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WO2006005683A1 - Phamaceutical preparations comprising insulin - Google Patents

Phamaceutical preparations comprising insulin Download PDF

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Publication number
WO2006005683A1
WO2006005683A1 PCT/EP2005/053070 EP2005053070W WO2006005683A1 WO 2006005683 A1 WO2006005683 A1 WO 2006005683A1 EP 2005053070 W EP2005053070 W EP 2005053070W WO 2006005683 A1 WO2006005683 A1 WO 2006005683A1
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WO
WIPO (PCT)
Prior art keywords
alkyl
aryl
independently selected
insulin
optionally substituted
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/EP2005/053070
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French (fr)
Inventor
Helle Birk Olsen
Niels Christian Kaarsholm
Peter Madsen
Per Balschmidt
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Novo Nordisk AS
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Novo Nordisk AS
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Filing date
Publication date
Application filed by Novo Nordisk AS filed Critical Novo Nordisk AS
Priority to EP05758689A priority Critical patent/EP1768694A1/en
Priority to JP2007519777A priority patent/JP2008505866A/en
Publication of WO2006005683A1 publication Critical patent/WO2006005683A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K33/00—Medicinal preparations containing inorganic active ingredients
    • A61K33/24—Heavy metals; Compounds thereof
    • A61K33/30—Zinc; Compounds thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/22—Hormones
    • A61K38/28—Insulins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P5/00—Drugs for disorders of the endocrine system
    • A61P5/48—Drugs for disorders of the endocrine system of the pancreatic hormones
    • A61P5/50—Drugs for disorders of the endocrine system of the pancreatic hormones for increasing or potentiating the activity of insulin

Definitions

  • the present invention discloses insulin preparations comprising ligands for the His B10 -Zn 2+ sites of the R-state insulin hexamer wherein the ligand is extended by protamine.
  • Insulin AHostery The insulin hexamer is an allosteric protein that exhibits both posi ⁇ tive and negative cooperativity and half-of-the-sites reactivity in ligand binding. This allosteric behaviour consists of two interrelated allosteric transitions designated L A 0 and L B 0 , three in ⁇ ter-converting allosteric conformation states (eq. 1),
  • T 61 T 3 R 3 and R 6 and two classes of allosteric ligand binding sites designated as the phenolic pockets and the His 610 anion sites. These allosteric sites are associated only with insulin subunits in the R conformation. insulin Hexamer Structures and Ligand Binding.
  • the T- to R-transition of the insulin hexamer involves transformation of the first nine residues of the B chain from an extended conformation in the T-state to an a-helical conformation in the R-state. This coil-to-heJix transition causes the N-terminal residue, Phe B1 , to undergo an ⁇ 30 A change in position.
  • This conformational change creates hydrophobic pockets (the phenolic pockets) at the sub- unit interfaces (three in T 3 R 3 , and six in R 8 ), and the new B-chain helices form 3-helix bun ⁇ dles (one in T 3 R 3 and two in R 6 ) with the bundle axis aligned along the hexamer three-fold symmetry axis.
  • the His 910 Zn 2+ in each R 3 unit is forced to change coordination geometry from octahedral to either tetrahedra! (monodentate ligands) or pentahedral (bidentate ligands).
  • Formation of the helix bundle creates a narrow hydrophobic tunnel in each R 3 unit that extends from the surface -12 A down to the His B1 ° metal ion. This tunnel and the His B1 ° Zn 2+ ion form the anion binding site.
  • absorption rates vary between about 1 hour (for rapid-acting insulin analogues, such as Asp B2 ⁇ human insulin) and about 4 hours (Co 3+ - hexamer).
  • Current Approaches Toward Slow Acting Insulins The inherent limitation of the ab ⁇ sorption half-life to about 4 hours for a soluble human insulin hexamer necessitates further modifications to obtain the desired protraction. Traditionally, this has been achieved by the use of preparations wherein the constituent insulin is in the form of a crystalline and/or amor ⁇ phous precipitate.
  • NPH and Ultratente belong to this category of insulin preparations where crystallization/precipitation is effected by the addition of protamine and excessive zinc ion, respectively.
  • the present invention provides insulin preparations comprising high-affinity ligands for the His Bi0 -Zn 2+ sites of the R-state insulin hexamer, zinc ions and insulin wherein the ligand is extended by protamine.
  • the resulting ligands with protamine extensions work to modify the time action pro ⁇ file of insulin formulations.
  • These preparations may be formulated with variable insulin spe- cies over a wide range of pH from 3.0 to 8.5 and their time action profiles may be tailored by suitable adjustments of anchor affinity as well as the concentration of protami ⁇ e-extended iigand.
  • the invention also provides a method of preparing ligands for the His 810 Zn 2+ sites of the R-state insulin hexamer comprising the steps of • Identifying a starter compound that binds to the R-state His B10 -Zn z+ site
  • Also provided are methods of treating type 1 or type 2 diabetes comprising adminis ⁇ tering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation of the invention.
  • Halogen designates an atom selected from the group consisting of F 1 Cl, Br and ⁇ .
  • C 1 -C 6 -BIkVl 11 as used herein represents a saturated, branched or straight hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n- pentyl, isopentyl, neopentyl, fert-pe ⁇ tyl, n-hexyl, isohexyl and the like.
  • CrCe-alkylene represents a saturated, branched or straight bivalent hydrocarbon group having from 1 to 6 carbon atoms.
  • C 2 -Ce-alkenyr represents a branched or straight hydro ⁇ carbon group having from 2 to 6 carbon atoms and at least one double bond.
  • groups include, but are not limited to. vinyl, 1-propenyl, 2-propenyl, iso-propenyl, 1,3- butadienyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyH-propenyl, 1-pentenyl, 2-pentenyl, 3- pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 2,4-hexadienyl, 5- hexenyl and the like.
  • C 2 -C 6 -alkynyr represents a branched or straight hydro ⁇ carbon group having from 2 to 6 carbon atoms and at least one triple bond.
  • groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2- butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pe ⁇ tynyl, 1-hexynyl, 2-hexynyl, 3- hexynyl. 4-hexynyl, 5-hexynyl, 2,4-hexadiynyl and the like.
  • d-Ce-alkoxy refers to the radical -O-d-C B -alkyl t wherein C r C 6 -alkyl is as defined above. Representative examples are methoxy, ethoxy, n-propoxy, iso- propoxy, butoxy, sec-butoxy, fert-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like.
  • C 3 -C 8 -cycloalkyr represents a saturated, carbocyclic group having from 3 to 8 carbon atoms. Representative examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.
  • d- ⁇ -cycloalkenyF represents a non-aromatic, carbocyclic group having from 4 to 8 carbon atoms containing one or two double bonds.
  • Representative examples are 1-cyclopente ⁇ yl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cycIohexenyl, 2-cyclo- hexenyl, 3-cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl, 2-cyclooctenyl, 1 ,4-cycioocta- dienyl and the like.
  • heterorocydyr as used herein represents a non-aromatic 3 to 10 membered ring containing one or more heteroatoms selected from nitrogen, oxygen and sulphur and op ⁇ tionally containing one or two double bonds.
  • Representative examples are pyrrolidinyl, piperi- dyl, piperazinyl, morpholinyl, thiomorpholinyl, aziridinyl, tetrahydrofuranyl and the like.
  • aryl as used herein is intended to include carbocyclic, aromatic ring sys ⁇ tems such as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, carbocyclic, aromatic ring systems. Representative examples are phenyl, biphenylyl, naphthyl, anthra- cenyl, phenanthrenyl, fluorenyl, indenyl, a ⁇ lenyl and the like. Any! is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4- dihydronaphthyl and the like.
  • arylene as used herein is intended to include divalent, carbocyclic, aro-
  • ring systems such as 6 membered monocyclic and 9 to 14 raembered bi- and tricyclic, divalent, carbocyclic, aromatic ring systems.
  • Representative examples are phenylene, bi- phenylylene, naphthylene, anthracenylene, phenanthrenylene, fluorenylene, indenylene, az- ulenylene and the like.
  • Arylene is also intended to include the partially hydrogenated deriva- fives of the ring systems enumerated above.
  • Non-limiting examples of such partially hydro ⁇ genated derivatives are 1 ,2,3,4-tetrahydronaphthyle ⁇ e, 1,4-dihydronaphthylene and the like.
  • aryloxy as used herein denotes a group -O-aryl, wherein aryl is as defined above.
  • aroyl denotes a group -C(O)-aryl, wherein aryl is as defined above.
  • heteroaryl as used herein is intended to include aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sul ⁇ phur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxy- gen and sulphur.
  • Representative examples are furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyra- zolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1 ,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1 ,2,4-triazinyl, 1 ,3,5- triazinyl, 1 ,2,3-oxadiazolyl.
  • Heteroaryl is also intended to include the partially hydrogenated de ⁇ rivatives of the ring systems enumerated above.
  • Non-limiting examples of such partially hy ⁇ drogenated derivatives are 2,3-dihydroben ⁇ ofuranyl, pyrrolinyl, pyrazoltnyl, indolinyl, oxazolid- tnyl, oxazo ⁇ nyl, oxazepinyl and the like.
  • heteroarylene as used herein is intended to include divalent, aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxy ⁇ gen and sulphur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitro- gen, oxygen and sulphur.
  • carbazolylene azepinylene, diazepinylene, acridinylene and the like.
  • Heteroaryl is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above.
  • Non-limiting examples of such partially hydrogenated deriva ⁇ tives are 2,3-dihydrobenzofuranylene, pyrrolinylene, pyrazolinylene, indolinylene, oxazolid- inylene, oxazolinylene, oxazepinylene and the like.
  • ArGI as used herein is intended to include an aryl or arylene radical as ap- plicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, anthracenyl, phena ⁇ threnyl, fluorenyl, indenyl, and azulenyl as well as the co ⁇ esponding divalent radicals.
  • ArG2 as used herein is intended to include an aryl or arylene radical as ap ⁇ plicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, fluorenyl, and indenyl, as well as the corresponding divalent radicals.
  • Hetr as used herein is intended to include a heteroaryl or heteroarytene radical as applicable, where heteroaryl or heteroarytene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1 ,2,3-triazinyi, 1 ,2,4-triazinyl, 1 ,3,5- triazinyl, 1 ,2.3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1 ,2,5-oxa- diazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadia
  • ⁇ et2 as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl. pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1 ,2,4-triazinyl.
  • Het3 is intended to include a heteroaryl or Heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl.
  • Aryl-CrCe-alkyl "heteroaryl-C r C 6 -alkyl”, “aryl-C ⁇ -C ⁇ -alkenyP etc. is intended to mean Ci-C 8 -alkyl or C 2 -C ⁇ -alkenyl as defined above, substituted by an aryl or heteroaryl as defined above, for example:
  • treatment means the management and care of a patient for the purpose of combating a disease, disorder or condition.
  • the term is intended to include the delaying of the progression of the disease, disorder or condition, the alleviation or relief of symptoms and complications, and/or the cure or elimination of the dis ⁇ ease, disorder or condition.
  • the patient to be treated is preferably a mammal, in particular a human being.
  • fragment as used herein is intended to mean a bivalent chemical group.
  • neutral amino acid as used herein is intended to mean any natural (cod- able) and non-natural amino acid, including ⁇ - or ⁇ -aminocarboxylic acids, including D- isomers of these (when applicable) without charges at physiologically relevant pH in the side chain, such as glycine, alanine, ⁇ -alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, aspargine, glutamine, cysteine, methionine, 3-aminobenzoic acid, 4-aminobenzoic acid or the like.
  • positively charged group as used herein is intended to mean any phar ⁇ maceutically acceptable group that contains a positive charge at physiologically relevant pH, such as amino (primary, secondary and tertiary), ammonium and guanidino groups.
  • ⁇ amino acid as used herein is intended to mean mean any natural (cod- able) and non-natural ⁇ -aminocarboxylic acid, including D-isomers of these.
  • ⁇ amino acid as used herein is intended to mean any ⁇ -aminocarboxylic add, such as ⁇ -alanine, isoserine or the like.
  • desB30 as used herein is intended to mean meant a natural insulin B chain or an analogue thereof lacking the B30 amino acid residue.
  • amino acid residues are indicated in the three letter amino acid code or the one letter amino code.
  • BIOS amino acid residue in position 1 in the B chain of insulin or analogue thereof (counted from the N- terminal end) and the amino acid residue in position 1 in the A chain of insulin or analogue thereof (counted from the N-terminal end), respectively.
  • groups of compounds such as carboxylates, dithiocarboxylates, phe ⁇ olates, thiophenolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano-1,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, naphthoic acids and salicylic acids, these groups of compounds are in ⁇ tended to include also derivatives of the compounds from which the groups take their name.
  • insulin refers to all variants of insulin including human in ⁇ sulin, an analogue thereof, a derivative thereof and combinations of any of these, acid- stabilised insulin, fast/rapid acting insulin and long/stow/basal acting insulin.
  • human insulin refers to naturally produced insulin or re- combinantly produced insulin. Recombinant human insulin may be produced in any suitable host cell, for example the host cells may be bacterial, fungal (including yeast), insect, animal or plant cells.
  • insulin analogue as used herein is meant human insulin in which at least one amino acid has been deleted and/or replaced by another amino acid including non- codeable amino acids, or human insulin comprising additional amino acids, i.e. more than 51 amino acids, such that the resulting analogue possesses insulin activity.
  • insulin derivative refers to human insulin or an analogue thereof which has been chemically modified, i.e. at least one organic substitue ⁇ t is bound to one or more of the amino acids, e.g. by introducing a side chain in one or more positions of the insulin backbone.or by oxidizing or reducing groups of the amino acid residues in theJnsulin or by converting a free carboxylic group to an ester group or acylating a free amino group or a hydroxy group.
  • acid-stabilised insulin refers to an insulin analog that does not deamidate or dimerize at pH values below 7. Specifically, the analog cannot have Asn or Asp as a C-terminal residue.
  • fast/rapid acting insulin as used herein is meant any insulin having an onset of action after injection or any other form of administration faster or equal to that of soluble and neutral formulations of human insulin.
  • long/slow/basal acting insulin as used herein is intended to include insu ⁇ lin compounds such as protamine insulin, zinc insulin, protamine zinc insulin.
  • phenolic compound or similar expressions as used herein refers to a chemical compound in which a hydroxyl group is bound directly to a benzene or substituted benzene ring. Examples of such compounds include, but are not limited to, phenol, o-cresol, m-cresol and p-cresol.
  • Protamine as used herein refers to the generic name of a group of strongly basic proteins present in sperm cell nucleic in saltlike combination with nucleic acids. Commercially available protamines can be isolated from mature fish sperm and are usually obtained as the sulphate. The peptide composition of a specific protamine may vary depending of which fam- ily, genera or species of fish it is obtained from.
  • Protamine usually contains four major com ⁇ ponents, i.e. single-chain peptides containing about 30-32 residues of which about 21-22 are arginines.
  • the N-ternimal is praline for each of the four main components, and since no other amino groups are present in the sequence, chemical modification of protamine is expected to be homogenoues in this context.
  • protamines to be used together with insulin are obtained from e.g. salmon (salmine), rainbow trout (iridine), herring (clupeine), sturgeon (sturine) or Spanish mackerel (thynnine).
  • Protamine also refers to preparations comprising salts of the proteins.
  • an insulin derivative according to the invention is stated to be “soluble at physiological pH values” it means that the insulin derivative can be used for preparing injectable insulin compositions that are fully dissolved at physiological pH values.
  • Such favourable solubil ⁇ ity may either be due to the inherent properties of the insulin derivative alone or a result of a fa ⁇ vourable interaction between the insulin derivative and one or more ingredients contained in the vehicle.
  • physiologically relevant pH as used herein is intended to mean a pH of about 7.1 to 7.9.
  • Fig. 1 Change in plasma glucose level after subcutaneous injection of the prepara- tion: 0,6mM A21G, B28D human insulin, 0.3mM Zn2+, 3OmM phenol, 1.6% glycerol, 0.3mM 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyrylprotamine.
  • Fig. 2 4H3N-assay. UV/vis spectra resulting from a titration of hexameric insulin with the compound 3-hydroxy-2-naphthoic acid in the presence of 4-hydroxy-3-n ⁇ trobenzoic acid (4H3N). Inserted in the upper right comer is the absorbance at 444nm vs. the co ⁇ cen- tration of ligand
  • Fig. 3 Fluorescence spectra resulting from a titration of hexameric in ⁇ sulin with 5-(3-methoxybenzylidene)thiazolidine-2,4-dione in the presence of 5-(4- dimethylaminobenzylidene)thiazolidine-2 T 4-dione (TZD). Inserted in the upper right comer is the fluorescence at 460 nm vs. the concentration of ligand
  • the present invention is based on the discovery that the His 910 Zn +* ligand binding sites of the R-state insulin hexamer wherein the ligand is extended by protamine can be used to obtain an insulin preparation having prolonged action designed for flexible injection re ⁇ gimes including once-daily, based on insulin molecules of any kind.
  • the basic concept underlying the present invention involves reversible attachment of a ligand to the His B1 ° Zn 2+ site of the R-state hexamer.
  • a suitable ligand binds to the hexamer metal site with one end while at the other end, the anchor features a carboxylate group.
  • the carboxylate group is covalently bound via an amide link to protamine.
  • the anions currently used in insulin formulations as allosteric ligands for the R-state hexamers bind only weakly to the His B1 ° anion site.
  • the present inven ⁇ tion which is based on the discovery of suitable higher affinity ligands for these anion sites, provides ligands which are extended to modify timing via changes in hexamer solubility as outlined above.
  • the His B1Q Zn 2* site consists of a tunnel or cavity with a triangular-shaped cross- section that extends -12 A from the surface of the hexamer down to the His 810 Zn 2* ion.
  • the diameter of the tunnel varies along its length and, depending on the nature of the ligand oc ⁇ cupying the site, the opening can be capped over by the Asn 63 and Phe B1 side chains.
  • the walls of the tunnel are made up of the side chains of the amino acid residues along one face each of the three ⁇ -helices.
  • the side chains from each helix that make up the lining of the tunnel are Phe B1 , Asn 83 , and Leu Be . Therefore, except for the zinc ion, which is coordinated to three His B1 ° residues and is positioned at the bottom of the tunnel, the site is principally hydrophobic.
  • substituents on the ligand it may be possible for substituents on the ligand to make H-bonding interactions with Asn B3 and with the peptide linkage to Cys B7 .
  • the present invention originates from a search for compounds with suitable binding properties by using UV-visible and fluorescence based competition assays described herein which are based on the displacement of chromophoric ligands from the R-state His B10 -Zn 2+ site by the incoming ligand in question. These compounds will be referred to as "starter com ⁇ pounds" in the following. These assays are easily transformed into a high-throughput format capable of handling fibraries constructed around hits from the initial search of compound da ⁇ tabases. These starter compounds provide the starting point for the task of constructing a chemical handle that allows for attachment of the protamine group.
  • this chemical group can be attached (optionally using a spacer group using and synthesis procedures known to those skilled in the art) to a position on the starter compound remote from the Zn 2* -binding functionality.
  • the invention thus provides pharmaceutical preparation comprising
  • Zinc ions A ligand which binds reversibly to a HisB10 Zn 2+ site of an R-state hexamer and wherein the ligand is extended by covalent attachment to protamine, having the following general formula (I)
  • CGr is a chemical group which reversibly binds to a His B1 ° Zn 2+ site of an insulin hexamer
  • Lnk is a linker selected from
  • B 2 is a valence bond, C ⁇ C ⁇ -alkylene.
  • C 2 -C 1B -alkenylene, C r Ci 8 -alkynylene, arylene. heteroarylene, -C t -Ci ⁇ -alkyl-aryl-, -Crde-alkenyl-aryl-. -Cz-Ci ⁇ -alkynyl-aryl-. -C( O)-
  • arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR 6 , -C(O)H, OCOR 6 , -SO 2 , -CN, -CF 3 , -
  • Frg is a fragment consisting of O to 5 neutral ⁇ - or ⁇ -amino acids
  • the present invention also encompasses pharmaceutically acceptable salts of the present compounds.
  • Such salts include pharmaceutically acceptable acid addition salts, pharmaceutically acceptable metal salts, ammonium and alkylated ammonium salts.
  • Acid addition salts include salts of inorganic acids as well as organic acids. Representative exam ⁇ ples of suitable inorganic acids include hydrochloric, hydrobromic, hydroiodic, phosphoric, sulphuric, nitric acids and the like.
  • suitable organic acids include formic, acetic, trichloroacetic, trifluoroacetic, propionic, benzoic, cinnamic, citric, fumaric, gly- colic, lactic, maleic, malic, malonic, ma ⁇ delic, picric, pyruvic, succinic, methanesulfonic, ethanesulfonic, tartaric, ascorbic, pamoic, , ethanedisulfonic, gluconic, citraconic, aspartic, stearic, palmitic, glycolic, p-aminobenzoic, glutamic, benzenesulfonic, p-toluenesulfonic acids and the like.
  • compositions include the pharmaceutically acceptable salts listed in J. Pharm. Sci. 1977, 66, 2, which is incorporated herein by reference.
  • metal salts include lithium, sodium, potassium, magnesium salts and the like.
  • ammonium and alkylated ammonium salts include ammonium, methyl-, dimethyl-, trimethyl-, ethyl-, hydroxyethyl-, diethyl-, n-butyh sec-butyi-, tert-butyh tetramethylammonium salts and the like.
  • pharmaceutically acceptable acid addition salts are the hydrates, which the present compounds, are able to form.
  • the acid addition salts may be obtained as the direct products of compound synthe ⁇ sis, in the alternative, the free base may be dissolved in a suitable solvent containing the ap ⁇ intestinalte acid, and the salt isolated by evaporating the solvent or otherwise separating the salt and solvent.
  • the compounds of the present invention may form solvates with standard low mo ⁇ lecular weight solvents using methods well known to the person skilled in the art. Such sol ⁇ vates are also contemplated as being within the scope of the present invention.
  • CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiophe ⁇ olates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano-1,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids.
  • CGr is a chemical structure selected from the group consist- ing of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidin- ediones, 5-mercaptotetrazoles, or 4-cyano-1 ,2,3-triazoles.
  • R 1 and R 4 are independently selected from hydrogen or Ci-C ⁇ -alkyl
  • R 2 is hydrogen or Ci-C 6 -alkyl or aryl
  • R 1 and R 2 may optionally be combined to form a double bond
  • R 3 and R 6 are independently selected from hydrogen, halogen, aryl, C r C 6 -alkyl, or -C(O)NR 11 R 12 .
  • a and B are independently selected from d-C ⁇ -alkylene, arylene, aryl-CrC 6 -alkyl-, aryl-C 2 - C ⁇ -alkenyl- or heteroarylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 6 and the arylene or heteroarylene is optionally substituted with up to four substituents R ? , R 8 , R 9 , and R 10 .
  • a and R 3 may be connected through one or two valence bonds
  • B and R 5 may be connected through one or two valence bonds
  • R e is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 ,
  • R 7 , R 8 , R B and R 10 are independently selected from • hydrogen, halogen, -CN, -CH 2 CN 1 -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 . -OS(O) 2 CF 3 , -SCF 3 . -NO 2 , -OR 11 , -NR 11 R 12 , -SR 11 , -NR 11 S(O) 2 R 12 , -S(O) 2 NR 11 R 12 , -S(O)NR 11 R 12 .
  • Ci-C ⁇ -alkyl C ⁇ -C ⁇ -alkenyl or Cz-C ⁇ -alkynyl. each of which may optionally be substi ⁇ tuted with one or more substituents independently selected from R 13 ,
  • each cyclic moiety may optionally be substituted with one or more substitu ⁇ ents independently selected from R 1 *,
  • R 11 and R 12 are independently selected from hydrogen, OH, d-Cj-o-alkyl, aryl-Ci-C fl -alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 ; R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 13 is independently selected from halogen. -CN, -CF 3 , -OCF 3 , -OR 11 , -C(O)OR 11 , -NR 11 R 12 , and -C(O)NR 11 R 12 ,
  • R 1 " is independently selected from halogen, -C(O)OR 11 , -CH 2 C(O)OR 11 , -CH 2 OR 11 , -CN, - CF 3 . -OCF 3 , -NO 2 , -OR 11 . -NR 11 R 12 , S(O) 2 R 11 , aryl and d-C ⁇ -aikyl,
  • R 15 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -Od-C ⁇ -alkyl, -C(O)OC 1 -C 6 - alkyl, -COOH and -NH 2
  • R l ⁇ is independently selected from halogen, -C(O)OC 1 -Ce-SlKyI, -COOH, -CN, -CF 3 .
  • X O.
  • Y is -O- or -S-.
  • Y is -O-.
  • Crg is arylene optionally substituted with up to four substituents, R 7 ,
  • R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from ArG1 optionally substituted with up to four sub ⁇ stituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is phenylene or naphtylene optionally substituted with up to four substituents, R 7 , R 8 , R ⁇ , and R 10 which may be the same or different.
  • A is phenylene
  • A is heteroarylene optionally substituted with up to four substituents
  • R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from Het1 optionally substituted with up to four sirb- stituents, R 7 , R ⁇ , R 9 , and R 10 which may be the same or different.
  • A is selected from Het2 optionally substituted with up to four sub ⁇ stituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from Het3 optionally substituted with up to four sub ⁇ stituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is selected from the group consisting of indolylene, benzofu- ranylidene, quinolylene, furylene, thienylene, or pyrrolylene, wherein each heteroaryl may optionally substituted with up to four substitue ⁇ ts, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is be ⁇ zofuranylene optionally substituted with up to four substitu- ents R 7 , R B , R 9 , and R 10 which may be the same or different.
  • R 7 , R B , R 9 , and R 10 which may be the same or different.
  • A is carbazolylidene optionally substituted with up to four substitu- ents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • A is quinolylidene optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different. In another embodiment A is
  • A is indolylene optionally substituted with up to four substituents R 7 , R 8 , R ⁇ , and R 10 which may be the same or different.
  • R 7 , R 8 , R ⁇ , and R 10 which may be the same or different.
  • R 1 is hydrogen. In another embodiment R 2 is hydrogen.
  • R 1 and R 2 are combined to form a double bond.
  • R 3 is d-C ⁇ -alkyl, halogen, or C(O)NR 18 R 17 .
  • R 3 is C r C 6 -alkyl or C(O)NR 16 R 17 .
  • R 3 is methyl.
  • R 7 , R B , R 9 , and R 10 which may be the same or different.
  • R 4 is hydrogen.
  • R 5 is hydrogen.
  • R 6 is aryl.
  • R ⁇ is phenyl.
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • R 7 , R ⁇ , R 8 and R 10 are independently selected from
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 .
  • R 7 , R 8 , R 9 and R 10 are independently selected from
  • each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 .
  • R 7 , R 8 , R 9 and R 10 are independently selected from •hydrogen, halogen, -OR 11 , -Od-C ⁇ -alkyl-qojOR 11 , or -C(O)OR 11 ,
  • R 11 and R 12 are independently selected from hydrogen, Ci-Ca-alkyl, aryl or aryKVCe-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R ie ; R 11 and R 1Z when at ⁇ tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
  • R 11 and R 12 are independently selected from hydrogen, CrCaralkyl, aryl or aryl-CrC ⁇ -alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 16 .
  • R 11 and R 12 are independently selected from phenyl or phenyl-C r C 6 - alkyl
  • R 11 and R 12 are methyl.
  • R 13 is independently selected from halogen, CF 3 , OR 11 or NR 11 R 12 .
  • R 13 is independently selected from halogen or OR 11 .
  • R 13 is OR 11 .
  • R 14 is independently selected from halogen, -C(O)OR 11 , -CN, -CF 3 , -
  • R 14 is independently selected from halogen, -C(O)OR 11 , or -OR 11 .
  • R 15 is independently selected from halogen, -CN, -CF 3 , -C(O)OC 1 -C 6 - alkyl.and -COOH.
  • R 15 is independently selected from halogen or -C(O)OCi-C ⁇ -alkyl.
  • R t ⁇ is independently selected from halogen
  • R 19 is hydrogen or Ci-C ⁇ -alkyl
  • R 20 is hydrogen or Ci-C 6 -alkyl
  • D and F are a valence bond or CVC ⁇ -alkylene optionally substituted with one or more sub- stituents independently selected from R 72 ,
  • R 72 is independently selected from hydroxy. Ci-C ⁇ -alkyl, or aryl,
  • E is Ci-C ⁇ -alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is option ⁇ ally substituted with up to three substituents R 21 , R 22 and R 23 ,
  • G is Ci-C ⁇ -alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is op- tionally substituted with up to three substituents R 24 , R 25 and R 26 ,
  • R 17 , R 18 , R z ⁇ R 22 , R 23 , R 24 , R 25 and R 26 are independently selected from
  • R 29 which may optionally be substituted with one or more substituents independently se ⁇ lected from R 29 , •aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-C ⁇ -alkoxy, aryl-CrC ⁇ -alkyl.
  • R 27 and R 28 are independently selected from hydrogen.
  • C r C ⁇ -alkyl, aryl-d-Ce-alkyl or aryl, or R 27 and R 2 ⁇ when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 29 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 2r , and -NR 27 R 28 ,
  • R 30 is independently selected from halogen, -C(Q)OR 27 . -CM, -CF 3 , -OCF 3 . -NO 2 , -OR 27 , -NR 27 R za and d-C 8 -alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
  • D is a valence bond.
  • D is Ci-C 6 -alkylene optionally substituted with one or more hydroxy, d-C ⁇ -alkyl, or aryl.
  • E is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • E is arylene optionally substituted with up to three substituents inde ⁇ pendently selected from R 21 , R 22 and R 23 .
  • E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • E is phe ⁇ ylene optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • CGr is
  • R 21 , R 22 and R 23 are independently selected from
  • R 21 , R 22 and R 23 are independently selected from
  • Ci-C ⁇ -alkyl optionally substituted with one or more substituents independently se ⁇ lected from R 29
  • R 21 , R 22 and R 23 are independently selected from
  • R 21 , R 22 and R 23 are independently selected from
  • R 21 , R 22 and R 23 are independently selected from
  • R 19 is hydrogen or methyl.
  • R 1 ⁇ is hydrogen.
  • R 87 is Hydrogen, CrC ⁇ -aIkyl or aryl.
  • R 27 is hydrogen or CrC ⁇ -alkyl.
  • R 28 is hydrogen or d-Cs-alkyl.
  • F is a valence bond.
  • F is C r C ⁇ -alkylene optionally substituted with one or more hydroxy, Ci-C ⁇ -alkyl, or aryl.
  • G is CrC ⁇ -alkylene or arylene, wherein the arylene is optionally sub ⁇ stituted with up to three substituents R 24 , R 25 and R 28 .
  • G is Ci-C 6 -alkylene or ArG1, wherein the arylene is optionally substi ⁇ tuted with up to three substituents R 24 , R 28 and R 2 ⁇
  • G is Ci-C ⁇ -alkylene.
  • G is phenylene optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • R 24 , R 25 and R 26 are independently selected from
  • R 24 , R 25 and R 26 are independently selected from
  • R 24 , R 25 and R 26 are independently selected from
  • R 24 , R 25 and R 26 are independently selected from
  • R 24 , R 2S and R 28 are independently selected from
  • R 24 , R 25 and R 28 are independently selected from
  • ArG1-Ci-C 6 -alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se ⁇ lected from R 30 .
  • R 20 is hydrogen or methyl.
  • R 20 is hydrogen
  • R 27 is hydrogen, Ci-C 6 -alkyl or aryl.
  • R 27 is hydrogen or C r C 6 -alkyl or ArG1. In another embodiment R 27 is hydrogen or C 1 -C ⁇ -alkyl.
  • R 28 is hydrogen or d-Ce-alkyl.
  • R 17 and R 18 are independently selected from
  • R 17 and R 18 are independently selected from •hydrogen, halogen, -CN. -CF 3 , -NO 2 , -OR 27 , -NR 27 R 28 , or -C(O)OR 27 ,
  • R 17 and R 1 ⁇ are independently selected from
  • R 17 and R 18 are independently selected from
  • R 17 and R 18 are independently selected from • hydrogen, halogen, -CN, -CF 3 , -NO 2 , -OR 27 . -NR 27 R 28 , or -C(O)OR 27
  • R 27 is hydrogen or C,-C ⁇ -alkyl. In another embodiment R 27 is hydrogen, methyl or ethyl. In another embodiment R 28 is hydrogen or Ci-C 6 -alkyl. In another embodiment R 28 is hydrogen, methyl or ethyl. In another embodiment R 72 is -OH or phenyl. In another embodiment CGr is
  • I is selected from *a valence bond
  • Z 1 is S(O) 2 or CH 2 .
  • Z 2 is -NH-, -O-or -S-, and n is 1 or 2,
  • Arylene -aryloxy-. arylene-oxycarbonyl-, -aroyl, arylene-C r C ⁇ -alkoxy-, ary- lene-CrC ⁇ -alkylene, arylene-C r C 6 -alkenylene, arylene-C 2 -C a -alkynylene, heteroary- len ⁇ , heteroarylene-Ca-Ce-alkenylene or heteroary- le ⁇ e-C 2 -C 6 -alkynylene, wherein the cyclic moieties are optionally substituted with one or more substituents selected from R 37 , R 31 is independently selected from hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF3, -OCHF 2 , -OCH 2 CF 3 .
  • R 32 and R 33 are independently selected from hydrogen, CrC ⁇ -alkyl or Ci-Ce-alkanoyl,
  • R 34 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR 35 R 38 ,
  • R 35 and R 36 are independently selected from hydrogen, CrC e -alkyl, aryl-d-C ⁇ -alkyl or aryl, or R 35 and R 36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 37 is independently selected from halogen, -C(OpR 35 , -C(O)H, -CN, -CF 3 , -OCF 3 , -NO 2 , - OR 35 . -NR 35 R 39 , d-Ce-alkyl or d-C ⁇ -alkanoyl.
  • t is a valence bond, -CH 2 N(R 32 )-, or -SO 2 N(R 33 )-.
  • 1 is a valence bond.
  • J is
  • J is «ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R 37 .
  • tn another embodiment J is
  • R 32 and R 33 are independently selected from hydrogen or Ci-C ⁇ -alkyl.
  • R 3 * is hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -SCF 3 , -NO 2 , -OR 35 ,
  • Ce-BlKyI-C(O)OR 35 -Sd-C 6 -alkyl-C(O)OR 35 or -C(O)OR 35 .
  • R 34 is hydrogen, halogen, -CF 3 . -NO 2 , -OR 35 , -NR 35 R 36 , -SR 35 , -NR 35 C(O)R 38 , or -C(O)OR 35 .
  • R 34 is hydrogen, halogen, -CF 3 , -NO 2 , -OR 35 , -NR 35 R 36 , or
  • R 34 is hydrogen, halogen, or -OR 35 .
  • R 33 and R 36 are independently selected from hydrogen, CrC ⁇ -alkyl, or aryl.
  • R 35 and R 36 are independently selected from hydrogen or C
  • R 37 is halogen. -C(O)OR 35 , -CN. -CF 3 , -OR 35 , -NR 35 R 36 , Ci-C e -alkyl or
  • R 37 is halogen, -C(O)OR 35 , -OR 35 , -NR 35 R 38 , d-C ⁇ -alkyl or C 1 -C 6 - alkanoyl.
  • R 37 is halogen, -C(O)OR 35 or -OR 35 .
  • U is a valence bond, d-Ce-alkenylene, -C t -C 6 -alkyl-O- or Ci-C ⁇ -alkyJe ⁇ e wherein any C,- Ce-alkyl moiety is optionally substituted with d-C ⁇ -alkyl,
  • R 38 is CrC ⁇ -alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R 39 ,
  • R 3B is independently selected from halogen, cyano, nitro, amino,
  • M is a valence bond, arylene or heteroaryfene, wherein (he aryl or heleroaryl moieties are optionally substituted with one or more substituents independently selected from R* 0 ,
  • R 40 is selected from
  • R 41 and R 42 are independently selected from hydrogen, -OH 1 d-C ⁇ -alkyl f d-C B -alkenyl, aryl- d-C ⁇ -alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R 45 , and the aryl moieties may optionally be substi- tuted with one or more substituents independently selected from R 46 ; R 41 and R 42 when at ⁇ tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 43 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 41 , and -NR 41 R 42
  • R 44 is independently selected from halogen, -C(O)OR 41 , -CH 2 C(O)OR 41 , -CH 2 OR 41 , -CN, - CF 3 . -OCF 3 , -NO 2 .
  • R 45 is independently selected from halogen. -CN, -CF 3 , -OCF 3 . -O-C-Ce-alkyl, -C(O)-O-C 1 - C ⁇ -alkyl, -COOH and -NH 2 ,
  • R 47 and R 48 are independently selected from hydrogen, C r C e -alkyl, aryl optionally substituted with one or more R 49 ,
  • R 49 is independently selected from halogen and -COOH
  • T is •CrC ⁇ -alkylene, C r C ⁇ -alkenylene , C 2 -C ⁇ -alkynylene, -CrC ⁇ -alkyloxy-carbonyl.
  • the alkylene, alkenylene and alkynylene moieties are optionally substituted with one or more substituents independently selected from R 50 , •arylene, -aryloxy-, -aryloxy-carbonyl-, arylene-d-C ⁇ -alkylene, -aroyl-, arylene-Ci- C ⁇ -alkoxy-, arylene-CrC ⁇ -alkenylene, arylene-C 2 -C e -alkynylene, heteroarylene, het- eroarylene-Ci-C ⁇ -alkylene, heteroarylene-C 2 -Ce-alkenylene, heteroarylene-C 2 - C ⁇ -alkynylene,
  • any alkylene, alkenylene , alkynylene. arylene and heteroarylene moiety is optionally substituted with one or more substituents independently selected from R 50 ,
  • R 51 and R 52 are independently selected from hydrogen and d-Ce-alkyl
  • R 53 is independently selected from Ci-C 6 -alkyl. d-Ce-alkoxy, -Ci-C 6 -alkyl-COOH, -Cr
  • K is a valence bond.
  • K is a valence bond.
  • CrC ⁇ -alkyl moiety is optionally substituted with R 38 .
  • K is a valence bond or d-C ⁇ -alkylene, wherein any d-C ⁇ -alkyl moi- ety is optionally substituted with R 38 .
  • K is a valence bond
  • U is a valence bond or -C,-C e -alkyl-O-.
  • U is a valence bond
  • M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • M is ArG1 or Het1, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • M is ArGI or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • M is phenylene optionally substituted with one or more substituents independently selected from R 40 .
  • M is indolylene optionally substituted with one or more substituents independently selected from R 40 .
  • M is
  • M is carbazolylene optionally substituted with one or more substitu ⁇ ents independently selected from R 40 .
  • M is
  • R 40 is selected from
  • Ci-C ⁇ -alkyl or C 2 -C 6 - alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 ,
  • R 40 is selected from
  • R 40 is selected from
  • R 41 and R 42 are independently selected from hydrogen, d-Ce-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
  • R 41 and R 42 are independently selected from hydrogen, methyl, ethyl. or phenyl, wherein the phenyl moieties may optionally be substituted with hafogen or -
  • Q is a valence bond, -CH 2 -. -CH 2 -CH 2 -, -CH 2 -O-, -CH 2 -CH 2 -O-,
  • R 4r and R 48 are independently selected from hydrogen, methyl and phenyl.
  • T is #Ci-C ⁇ -alkylene optionally substituted with one or more substituents independently selected from R 50 ,
  • ArGI 1 ArGI-d-C ⁇ -alkylene, Het3, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R w .
  • Ci-Ce-alkylene optionally substituted with one or more substituents independently selected from R 50 ,
  • R 50 is d-C ⁇ -alkyl, d-C ⁇ -alkoxy, aryl, aryloxy, aryl-d-C ⁇ -alkoxy , -OR 61 , -NO 2 , halogen, -COOH 1 -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 .
  • R 50 is d-Ce-alkyl, aryloxy, aryl-Ci-C 6 -alkoxy , -OR 51 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 .
  • R 50 is C r C 8 -alkyl, ArGI-O-, ArG1-C r Ce-alkoxy , -OR 51 , halogen, -COOH, -CF 3 , wherein any aryl moiety is optionally substituted with one or more R 53 .
  • R 50 is phenyl, methyl or ethyl.
  • R 50 is methyl or ethyl.
  • R 51 is methyl.
  • R 53 is d-Ce-alkyl, d-C 6 -alkoxy, -OR 51 , halogen.or -CF 3 .
  • V is Ci-C ⁇ -a1kylene, arylene, heteroarylene, arylene-Ci-e-alkylene or aryfene-C 2 ⁇ - alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 55 ,
  • R 54 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2
  • R 55 is independently selected from -hydrogen, halogen. -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 ,
  • -OS(O) 2 R » -C(O)NR 56 R 57 , -OC(O)NR 50 R 57 , -NR 56 C(O)R 57 , -CH 2 C(O)NR 56 R 57 . -OC 1 -C 6 - 8 ⁇ yI-C(O)NR 56 R 57 , -CH 2 OR 56 , -CH 2 OC(O)R 58 , -CH 2 NR 56 R 57 . -OC(O)R 58 , -OC 1 -C 8 - alkyl-CtOJOR 59 , -SCrC ⁇ -alkyl-CCOJOR 56 , -Cz-C ⁇ -alkenyl-
  • R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 58 is independently selected from hafoge ⁇ , -CN, -CF 3 , -OCF 3 , -OR 56 , and -NR 56 R 57 ,
  • R 59 is independently selected from halogen. -C(O)OR 56 . -CH 2 C(O)OR 56 , -CH 2 OR 59 , -CN, - CF 3 , -OCF 3 , -NO 2 , -OR 56 , -NR 56 R 57 and d-Qralkyl,
  • R 62 is d-Ce-alkyl, aryl optionally substituted with one or more substituents independently se ⁇ lected from halogen, or heteroaryl optionally substituted with one or more C r C ⁇ -alkyl inde ⁇ pendently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
  • V is arylene, heteroarylene, or arylene-Ci.C ⁇ -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected R 54 , and the arylene or heteroarylene is optionally substituted with one or more substituents inde- pendently selected from R 55 .
  • V is arylene, Het1, or aryle ⁇ e-d.C ⁇ -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde ⁇ pendently selected from R 55 .
  • V is arylene, Het2, or arylene-d-C ⁇ -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde ⁇ pendently selected from R 55 .
  • V is arylene, Het3, or arylene-d.C ⁇ -alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R 54 , and the arylene or heteroarylene moiety is optionally substituted with one or more substituents Inde ⁇ pendently selected from R 55 .
  • V is arylene optionally substituted with one or more substituents in ⁇ dependently selected from R 55 .
  • V is ArG1 optionally substituted with one or more substituents inde ⁇ pendently selected from R 53 .
  • V is phenylene, naphthyle ⁇ e or anthranylene optionally substituted with one or more substituents independently selected from R 55 .
  • V is phenylene optionally substituted with one or more substituents independently selected from R 55 .
  • R 55 is independently selected from
  • Ci-C ⁇ -alkyl optionally substituted with one or more substituents independently se- lected from R 58
  • R 55 is independently selected from • halogen, d-C ⁇ -alkyl, -CN. -OCF 3 ,-CF 3 . -NO 2 , -OR 56 , -NR 58 R 57 . -NR 58 C(O)R 57
  • Ci-Ce-alkyl optionally substituted with one or more substituents independently se ⁇ lected from R 58
  • R 55 is independently selected from halogen, -OR 59 , -NR 58 R 57 , -C(O)OR 58 , -OCrC-alkyl-CfOJOR 56 , -NR 58 C(O)R 57 or C r C ⁇ -alkyl.
  • R 55 is independently selected from halogen, -OR 56 , -NR 58 R 57 . -C(O)OR 58 , -OC-C ⁇ -alkyl-qojOR 56 , -NR 58 C(O)R 57 , methyl or ethyl.
  • R 56 and R S7 are independently selected from hydrogen or d-C 12 -alkyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 mem- bered heterocyclic ring with the said nitrogen atom.
  • R 56 and R 57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • AA is Ci-C ⁇ -alkylene, arylene, heteroarylene, arylene-Ci-C 6 -alkylene or arylene-Cj.
  • C ⁇ -alkenylene wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R 63 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 84 ,
  • R 63 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 ,
  • R 6 * is independently selected from
  • R 67 is independently selected from halogen. -CN, -CF 3 . -OCF 3 , -OR 65 , and -NR 65 R 66 .
  • R 68 is independently selected from halogen, -C(O)OR 65 . -CH 2 C(O)OR 65 , -CH 2 OR 65 , -CN, - CF 3 , -OCF 3 , -NO 2 , -OR 65 , -NR 65 R 68 and d-Ce-alkyl.
  • R 69 is independently selected from Ci-C ⁇ -alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more Ci-C ⁇ -alkyl,
  • R 70 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OC r C ⁇ -afkyl, -C(O)OC 1 -Ce- alkyl, -COOH and -NH 2 ,
  • R 71 is independently selected from halogen, -C(O)OCi-C ⁇ -alkyl, -COOH, -CN, -CF 3 , -OCF 3 , -
  • AA is arylene, heteroarylene or arylene-d.C ⁇ -alkylene, wherein the alkylene is optionally substituted with one or more R 63 , and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R 64 .
  • AA is aryle ⁇ e or heteroarylene, wherein the aryiene or heteroarylen ⁇ is optionally substituted with one or more substituents independently selected from R 64 .
  • AA is ArG1 or Het1 optionally substituted with one or more substitu ⁇ ents independently selected from R 64 .
  • AA is ArG1 or Het2 optionally substituted with one or more substitu ⁇ ents independently selected from R 84 .
  • AA is ArG 1 or Het3 optionally substituted with one or more substitu ⁇ ents independently selected from R 64 .
  • AA is phenylene, naphtylene, anthrylene, carbazolylene, thienylene, pyridylene, or benzodioxylene optionally substituted with one or more substituents independ ⁇ ently selected from R 64 .
  • AA is phenylene or naphtylene optionally substituted with one or more substituents independently selected from R 64 .
  • R 64 is independently selected from hydrogen, halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , C-Ce-alkyl, -OC(O)R 65 , -OC 1 -C 6 -BlRyI-C(O)OR 65 , aryloxy or aryl, wherein Ci-C B -alkyl is optionally substituted with one or more substituents independ ⁇ ently selected from R 97 , and the cyclic moieties optionally are substituted with one or more substituents independently selected from R 88 .
  • R 84 is independently selected from halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 66 , methyl, ethyl, propyl, -OC(O)R 65 , -OCH 2 -C(O)OR 65 , -OCH 2 -CH 2 -C(O)OR 65 , phenoxy optionally substituted with one or more substituents independently selected from R 68 .
  • R 65 and R eB are independently selected from hydrogen, CF 3 , C ⁇ C ⁇ -alkyl, aryl, or heteroaryl optionally substituted with one or more substituents inde- pendently selected from R 71 .
  • R 65 and R 8 ⁇ are independently hydrogen, Ci-C 1z -alkyl, aryl, or het ⁇ eroaryl optionally substituted with one or more substituents independently selected from R 7 ⁇
  • R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents inde- pendently selected from R 71 .
  • R 65 and R ⁇ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents inde ⁇ pendently selected from R 71 .
  • R 65 and R ⁇ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG 1 or Het3 optionally substituted with one or more substituents inde ⁇ pendently selected from R 71 .
  • R 65 and R e ⁇ are independently hydrogen, methyl, ethyl, propyl, butyl,
  • R 71 is halogen or Ci-C ⁇ -alkyl.
  • R 71 is halogen or methyl.
  • Frg consists of 0 to 5 neutral amino acids independently selected from the group consisting of GIy, Ala, Thr, and Ser.
  • Frg consists of 0 to 5 GIy. In another embodiment Frg consists of O GIy.
  • Frg consists of 1 GIy.
  • Frg consists of 2 GIy.
  • Frg consists of 3 GIy.
  • Frg consists of 4 GIy. In another embodiment Frg consists of 5 GIy.
  • G B is of the formula B'-B ⁇ CKO ⁇ -, B'-B ⁇ SOz- or B 1 ⁇ -CH 2 -, wherein
  • G B is of the formula B 1 -B 2 -C(O)-.
  • G B is of the formula B 1 -B 2 -C(O)-, B ⁇ B 2 OH 2 - or B T -B 2 -NH- t wherein
  • G B is of the formula B 1 -B 2 -CH 2 -, B 1 -B 2 -S ⁇ 2- or B 1 -B 2 -NH-. wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B 1 -B 2 -C(O)- or B 1 ⁇ -SO 2 -, wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B 1 ⁇ -C(O)- or B 1 -B 2 -CH 2 -, wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B 1 -B 2 -C(O)- or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B n -B 2 -CH 2 - or B 1 -B 2 -SO 2 - , wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B 1 -B 2 -NH- or B 1 -B 2 -SO 2 - , wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B'-B ⁇ CHz- or B 1 -B 2 -NH- , wherein B 1 and B 2 are as defined in claim 1.
  • G B is of the formula B 1 -B 2 -C(O)-. In another embodiment G B is of the formula B 1 -B 2 -CH 2 -. In another embodiment G B is of the formula B 1 -B 2 -SO 2 -. In another embodiment G B is of the formula B 1 -B 2 -NH-. In another embodiment B 1 is a valence bond, -O-, or -S-. In another embodiment B 1 is a valence bond, -O-. or -N ⁇ R 6 )-. In another embodiment B 1 is a valence bond, -S-, or -N(R 6 )-.
  • B 1 is -O-, -S- or -N(R 6 )-.
  • B 1 is a valence bond or -O-.
  • B 1 is a valence bond or -S-.
  • B 1 is a valence bond or -N(R 6 )-.
  • B 1 is -O-or -S-.
  • B 1 is -O-or -N(R 6 )-.
  • B 1 is -S-or -N(R 6 )-.
  • tn another embodiment B 1 is a valence bond.
  • B 1 is -O-.
  • B 1 is -S-.
  • B 1 is -N(R 8 )-.
  • B 2 is a valence bond, d-C t ⁇ -alkylene, C 2 -Ci a -alkenylene, C 2 -Ci 8 - alkynylene. arylene, heteroarylene.
  • B 2 is a valence bond, Ci-d 8 -alkylene, arylene, heteroarylene, -Ci- Ci 8 -alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
  • B 2 is a valence bond, d-Ci ⁇ -alkylene, arylene, -d-C ⁇ alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
  • B 2 is a valence bond or and the alkylene moieties are optionally substituted as defined in claim 1.
  • the insulin is selected from the goup consisting of human in ⁇ sulin, an analogue thereof, a derivative thereof and combinations of any of these.
  • the insulin is human insulin.
  • the insulin is an analogue of human insulin.
  • the insulin is a derivative of human insulin. In another embodiment the insulin is an analogue of human insulin wherein position
  • B28 is Asp, GIu, Lys, Leu, VaI, or Ala.
  • the insulin is an analogue of human insulin wherein position B28 is Asp, GIu or Lys
  • the insulin is an analogue of human insulin wherein position B28 is Asp or GIu.
  • the insulin is an analogue of human insulin wherein position B28 is Asp.
  • the insulin is an analogue of human insulin wherein position B28 is GIu. In another embodiment the insulin is an analogue of human insulin wherein position B28 is GIu. In another embodiment the insulin is an analogue of human insulin wherein position B28 is GIu.
  • B29 is Pro, Asp or GlU/
  • the insulin is an analogue of human insulin wherein position B29 is Pro or GIu.
  • the insulin is an analogue of human insulin wherein position B29 is Pro.
  • the insulin is an analogue of human insulin wherein position B29 is GIu.
  • the insulin is an analogue of human insulin wherein position
  • B9 is Asp or GIu.
  • the insulin is an analogue of human insulin wherein position B10 is Asp or GIu.
  • the insulin is an analogue of human insulin wherein position B10 is GIu. In another embodiment the insulin is an analogue of human insulin wherein position B1 is GIy.
  • the insulin is an analogue of human insulin wherein position 83 is Lys, Thr, Ser, Ala or GIn. In another embodiment the insulin is an analogue of human insulin wherein position 83 is Lys, Thr, Ser, Ala or GIn. In another embodiment the insulin is an analogue of human insulin wherein position 83 is Lys, Thr, Ser, Ala or GIn. In another embodiment the insulin is an analogue of human insulin wherein position
  • B3 is Lys, Thr, Ser or Ala.
  • the insulin is an analogue of human insulin wherein position B3 is Lys or Ala.
  • the insulin is an analogue of human insulin wherein position B3 is Lys.
  • the insulin is an analogue of human insulin wherein position B3 is Lys and position B29 is GIu.
  • the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin
  • the insulin is an analogue of human insulin wherein position B30 is deleted.
  • the insulin is an analogue of human insulin wherein position A18 is Gln.
  • the insulin is an analogue of human insulin wherein position A21 is Ala, GIn, GIu, GIy, His, lie, Leu, Met, Phe, Ser, Thr, Trp, Tyr, VaI or hSer.
  • the insulin is an analogue of human insulin wherein position A21 is Ala, GIy, lie, Leu, Phe, Ser, Thr, VaI or hSer. In another embodiment the insulin is an analogue of human insulin wherein position A21 is Ala, GIy, lie, Leu, Phe, Ser, Thr, VaI or hSer. In another embodiment the insulin is an analogue of human insulin wherein position A21 is Ala, GIy, lie, Leu, Phe, Ser, Thr, VaI or hSer. In another embodiment the insulin is an analogue of human insulin wherein position
  • A21 is Ala or GIy.
  • the insulin is an analogue of human insulin wherein position A21 is GIy.
  • the insulin is a derivative of human insulin or an analogue thereof having one or more lipophilic substituents.
  • the insulin is a derivative of human insulin or an analogue thereof wherein the N f -amino group in position B29Lys is modified by covalent acylation with a hydrophobic moiety such as an fatty acid derivative or an litocholic acid derivative.
  • the insulin derivative is selected from the group consisting of B29-N ⁇ -myristoyI ⁇ des(B30) human insulin, B29-N E -palmitoyl-des(B30) human insulin, B29-N ⁇ - myristoyl human insulin, B29-N ⁇ -palmitoyl human insulin, B28-N ⁇ -myristoyl Lys B2S Pro 829 hu- man insulin.
  • analogs of human insulin contain any combination of the additional stabilizing substitutions in positions B1, B3, A18 and A21.
  • the insulin is an analogue of human insulin selected from the group consisting of:
  • the insulin is an analogue of human insulin selected from the group consisting of:
  • A21G, B10E,desB30 A21G, desB25. desB30.
  • B29E B1 G, A21 G, B3K, B29E, desB30
  • the insulin is an analogue of human insulin from above three lists further modified in positions B3 and A18, eg B3T, B3S. B3Q and A18Q.
  • the insulin is an analogue of human insulin from the above three lists further modified as follows: B3T, B28D B3T, desB27.
  • the insulin is an analogue of human insulin from the above three lists further modified by deletion of B30.
  • the ratio of the protamine-exte ⁇ ded ligand of general for ⁇ mula (I) to zinc ion is 1:20 to 20:1.
  • the ratio of the protami ⁇ e-extended ligand of general for ⁇ mula (I) to zinc ion is 1:6 to 10:1.
  • the amount of zinc ions is 2-6 moles per mole of putative in- sulin hexamer.
  • the amount of zinc ions is 2.0-3.5 moles per putative insulin hexamer.
  • zinc ions are present in an amount corresponding to 10 to 40 ⁇ g Zn/100 U insulin. In another embodiment zinc ions are present in an amount corresponding to 10 to 26 ⁇ g Zn/100 U insulin.
  • the ratio between insulin and the protamine-extended ligand of the invention is in the range from 99:1 to 1:99.
  • the ratio between insulin and the protamine-extended ligand of the invention is in the range from 95:5 to 5:95.
  • the ratio between between insulin and the protamine- extended ligand of the invention is in the range from 80:20 to 20:80.
  • the ratio between between insulin and the protamine- extended ligand of the invention is in the range from 70:30 to 30:70.
  • the invention relates to a method of preparing a protamine- extended ligand of the invention comprising the steps of:
  • the invention relates to a method of prolonging the action of an in ⁇ sulin preparation which comprises adding the ligand of the invention to the insulin prepara ⁇ tion.
  • the invention in another aspect relates to a method of treating type 1 or type 2 dia- betes comprising administering to a patient in need thereof a theraputicaHy effective amount of a pharmaceutical preparation comprising
  • a ligand that binds to the R-state His B10 -Zn 2* site where said ligand may be as described in the embodiments above.
  • the invention provides an embodiment 1, which is a pharmaceuti ⁇ cal preparation comprising
  • a ligand which binds reversibly to a HisB10 Zn 2+ site of an R-state hexamer and wherein the ligand is extended by covalent attachment to protamine having the following general formula (I)
  • CGr is a chemical group which reversibly binds to a His 810 Zn 2+ site of an insulin hexamer
  • Lnk is a linker selected from
  • R 6B and R 7B are independently H, C r C 4 -alkyl;
  • Frg is a fragment consisting of O to 5 neutral ⁇ - or ⁇ -amino acids, or
  • CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano- 1 ,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thiazolidinediones, tetrazoles, 5- mercaptotetrazoles, rhodanines.
  • Embodiment 3 A pharmaceutical preparation according to embodiment 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2- ⁇ apthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or A- cyano-1 ,2,3-triazoles.
  • Embodiment 4 A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
  • Y is -S-, -O- or -NH-
  • R 1 , R 1A and R 4 are independently selected from hydrogen or Ci-C 6 -alkyl
  • R 2 and R 2 * are hydrogen or CrCe-alkyl or aryl, R 1 and R 2 may optionally be combined to form a double bond, R 1A and R 2 * may optionally be combined to form a double bond,
  • R 3 , R 3 * and R 5 are independently selected from hydrogen, halogen, aryl optionally substi- tuted with one or more substituents independently selected from R 16 .
  • d-Ce-alkyl, or -C(O)NR 11 R 12 , A, A 1 and B are independently selected from Ci-C 6 -alkyl, aryl, aryl-C r C fl -aIkyl, -NR 11 -ary[, aryI-C 2 -C 6 -alkenyl or heteroaryl, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R e and the aryl or heteroaryl is optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 , A and R 3 may be connected through one or two valence bonds, B and R 5 may be connected through one or two valence bonds,
  • R ⁇ is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 , R 7 , R 8 , R 9 and R 10 are independently selected from
  • each cyclic moiety may optionally be substituted with one or more substitu ⁇ ents independently selected from R 14 ,
  • R 11 and R 12 are independently selected from hydrogen, OH, Ci-C ⁇ -alkyl, aryl-C ⁇ Ce-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substituents independently selected from R 1 ⁇ ;
  • R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds
  • R 13 is independently selected from halogen, -CN. -CF 3 , -OCF 3 , -OR 11 , -C(O)OR 11 , -NR 11 R 12 , and -C(O)NR 11 R 12 ,
  • R 14 is independently selected from halogen, -C(O)OR 11 , -CH 2 C(O)OR 11 , -CH 2 OR 11 , -CN, - CF 3 , -OCF 3 , -NO 2 , -OR 11 , -NR 11 R 12 , -NR 11 C(O)R 11 . -S(O) 2 R 11 , aryl and C r C ⁇ -alkyl,
  • Embodiment 8. A pharmaceutical composition according to any one of the embodiments 4 to 7 wherein Y is -O- or -S-.
  • Embodiment 9 A pharmaceutical composition according to embodiment 8 wherein Y is -O-.
  • Embodiment 10 A pharmaceutical composition according to embodiment 8 wherein Y is
  • Embodiment 11 A pharmaceutical composition according to embodiment 8 wherein Y is -S-.
  • Embodiment 12 A pharmaceutical composition according to any one of the embodiments 4 to 11 wherein A is aryl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 13 A pharmaceutical composition according to embodiment 12 wherein A is selected from ArG1 optionally substituted with up to four substituents, R 7 , R 8 , R B , and R 10 which may be the same or different.
  • Embodiment 14 A pharmaceutical composition according to embodiment 13 wherein A is phenyl or naphtyl optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 15 A pharmaceutical composition according to embodiment 14 wherein A is
  • Embodiment 17 A pharmaceutical composition according to any one of the embodiments 4 to 11 wherein A is heteroaryl optionally substituted with up to four substitue ⁇ ts, R 7 . R 8 , R ⁇ , and R 10 which may be the same or different.
  • Embodiment 18 A pharmaceutical composition according to embodiment 17 wherein A is selected from Het1 optionally substituted with up to four substituents, R ⁇ , R ⁇ , R a , and R 10 which may be the same or different.
  • Embodiment 19 A pharmaceutical composition according to embodiment 18 wherein A is selected from Het2 optionally substituted with up to four substituents, R 7 t R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 20 A pharmaceutical composition according to embodiment 19 wherein A is selected from Het3 optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R t0 which may be the same or different.
  • Embodiment 21 A pharmaceutical composition according to embodiment 20 wherein A is selected from the group consisting of indolyl, benzofuranyl, quinolyl, furyl, thienyl, or pyrrolyl. wherein each heteroaryl may optionally substituted with up to four substituents, R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 22 A pharmaceutical composition according to embodiment 20 wherein A is benzofuranyl optionally substituted with up to four substituents R 7 , R 8 . R ⁇ , and R 10 which may be the same or different.
  • Embodiment 23 A pharmaceutical composition according to embodiment 22 wherein A is
  • Embodiment 26 A pharmaceutical composition according to embodiment 20 wherein A is quinolyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 27 A pharmaceutical composition according to embodiment 26 wherein A is
  • Embodiment 28 A pharmaceutical composition according to embodiment 20 wherein A is indolyl optionally substituted with up to four substituents R 7 , R 8 , R 9 , and R 10 which may be the same or different.
  • Embodiment 29 A pharmaceutical composition according to embodiment 28 wherein A is
  • Embodiment 30 A pharmaceutical composition according to any one of the embodiments 4 to 29 wherein R 1 is hydrogen.
  • Embodiment 31 A pharmaceutical composition according to any one of the embodiments 4 to 30 wherein R 2 is hydrogen.
  • Embodiment 32 A pharmaceutical composition according to any one of the embodiments 4 to 29 wherein R 1 and R 2 are combined to form a double bond.
  • Embodiment 33 A pharmaceutical composition according to any one of the embodiments 4 to 32 wherein R 3 is d-C ⁇ -alkyl, halogen, or C(O)NR 16 R 17 .
  • Embodiment 34 A pharmaceutical composition according to embodiment 33 wherein R 3 is
  • Embodiment 35 A pharmaceutical composition according to embodiment 34 wherein R 3 is methyl.
  • Embodiment 36 A pharmaceutical composition according to any one of the embodiments 4 to 11 wherein B is phenyl optionally substituted with up to four substituents, R 7 , R s , R 9 , and R 10 which may be the same or different.
  • Embodiment 37 A pharmaceutical composition according to any one of the embodiments 4 to 11 or 36 wherein R 4 is hydrogen.
  • Embodiment 38 A pharmaceutical composition according to any one of the embodiments 4 to 11 or 36 to 37 wherein R 5 is hydrogen.
  • Embodiment 39 A pharmaceutical composition according to any one of the embodiments 4 to 38 wherein R 6 is aryl.
  • Embodiment 40 A pharmaceutical composition according to embodiment 39 wherein R ⁇ is phenyl.
  • Embodiment 41 A pharmaceutical composition according to any one of the embodiments 4 to 40 wherein R 7 , R ⁇ , R ⁇ and R 10 are independently selected from
  • Embodiment 42 A pharmaceutical composition according to embodiment 41 wherein R 7 , R ⁇ , R 9 and R 10 are independently selected from
  • Ci-Ce-alkyl or CrC ⁇ -alkenyl which may each optionally be substituted with one or more substituents independently selected from R 13 • aryl, aryloxy, aroyl, aryl-Ci-C a -alkoxy, aryl-Ci-Ce-alkyl, heteroaryl,
  • each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 .
  • Embodiment 43 A pharmaceutical composition according to embodiment 42 wherein R 7 , R 8 , R B and R 10 are independently selected from
  • Embodiment 44 A pharmaceutical composition according to embodiment 43 wherein R 7 , R 8 , R 9 and R 10 are independently selected from
  • each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R 14 .
  • Embodiment 45 A pharmaceutical composition according to embodiment 44 wherein R 7 , R 8 , R 9 and R 10 are independently selected from
  • each of the cyclic moieties optionally may be substituted with one or more substitu- ents independently selected from R 14 .
  • Embodiment 46 A pharmaceutical composition according to embodiment 45 wherein R 7 , R 8 , R 9 and R 10 are independently selected from
  • Embodiment 47 A pharmaceutical composition according to any one of the embodiments 4 to 46 wherein R 11 and R 12 are independently selected from hydrogen, CrC 2 o-alkyl, aryl or aryl-d-C f i-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substi- tuted one or more substituents independently selected from R 1 ⁇ ; R 11 and R 12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitro ⁇ gen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
  • R 11 and R 12 are independently selected from hydrogen, CrC 2 o-alkyl, aryl or aryl-d-C f i-alkyl, wherein the alkyl groups may optionally be substituted with
  • Embodiment 48 A pharmaceutical composition according to embodiment 47 wherein R 11 and R 12 are independently selected from hydrogen, d-Czo-alkyl, aryl or aryl-C,-C ⁇ -alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R 15 , and the aryl groups may optionally be substituted one or more substitu ⁇ ents independently selected from R 1 ⁇ .
  • Embodiment 49 A pharmaceutical composition according to embodiment 48 wherein R 11 and R 12 are independently selected from phenyl or phenyl-Ci-C ⁇ -alkyl.
  • Embodiment 50 A pharmaceutical composition according to embodiment 48 wherein one or both of R 11 and R 12 are methyl.
  • Embodiment 51 A pharmaceutical composition according to any one of the embodiments 4 to 50 wherein R 13 is independently selected from halogen, CF 3 , OR 11 or NR 11 R 12 .
  • Embodiment 52 A pharmaceutical composition according to embodiment 51 wherein R 13 is independently selected from halogen or OR 11 .
  • Embodiment 53 A pharmaceutical composition according to embodiment 52 wherein R is
  • Embodiment 54 A pharmaceutical composition according to any one of the embodiments 4 to 53 wherein R 14 is independently selected from halogen, -C(O)OR 11 , -CN, -CF 3 , -OR 11 ,
  • Embodiment 55 A pharmaceutical composition according to embodiment 54 wherein R 14 is independently selected from halogen, -C(O)OR 11 , or -OR 11 .
  • Embodiment 56 A pharmaceutical composition according to any one of the embodiments 4 to 55 wherein R 15 is independently selected from halogen, -CN, -CF 3 , -CfOJOCrC ⁇ -alkyl.and
  • Embodiment 57 A pharmaceutical composition according to embodiment 56 wherein R 15 is independently selected from halogen or -C(O)OCi-C ⁇ -alkyl.
  • Embodiment 5 ⁇ A pharmaceutical composition according to any one of the embodiments 4 to 57 wherein R 1 ⁇ is independently selected from halogen, -C(O)OC r C a -alkyl, -COOH, -NO 2 ,
  • Embodiment 59 A pharmaceutical composition according to embodiment 58 wherein R 19 is independently selected from halogen, -C(O)OCi-C 6 -afkyl, -COOH, -NO 2 , or d-C 6 -alkyl.
  • Embodiment 60 A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
  • R 1 ⁇ is hydrogen or C r C ⁇ -alkyK
  • R 20 is hydrogen or d-C 6 -alkyl
  • D, D 1 and F are a valence bond, d-C ⁇ -alkylene or d-C 6 -alkenylene optionally substituted with one or more substituents independently selected from R 72 ,
  • R 72 is independently selected from hydroxy.
  • E is C t -C ⁇ -alkyl, aryl or h ⁇ teroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents R 21 , R 22 and R 23 ,
  • G and G 1 are d-C ⁇ -alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally sub ⁇ stituted with up to three substituents R 24 , R 25 and R 2 ⁇ ,
  • R 24 , R 25 and R 26 are independently selected from
  • R 27 and R 28 are independently selected from hydrogen, Ci-C 6 -alkyl, aryl-CrC ⁇ -alkyl or aryl, or R 27 and R 23 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 29 Is independently selected from halogen, -CN 1 -CF 3 , -OCF 3 .
  • -OR 27 , and -NR 27 R 28 , R 30 is independently selected from halogen, -C(O)OR 27 , -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 27 ,
  • Embodiment 61 A pharmaceutical composition according to embodiment 60 wherein D is a valence bond.
  • Embodiment 62 A pharmaceutical composition according to embodiment 60 wherein D is
  • C-pCe-alkylene optionally substituted with one or more hydroxy, C,-Ce-alkyl, or aryl.
  • Embodiment 63 A pharmaceutical composition according to any one of the embodiments 60 to 62 wherein E is aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • Embodiment 64 A pharmaceutical composition according to embodiment 63 wherein E is aryl optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • Embodiment 65 A pharmaceutical composition according to embodiment 64 wherein E is selected from ArG1 and optionally substituted with up to three substttuents independently selected from R 21 , R 22 and R 23 .
  • Embodiment 66 A pharmaceutical composition according to embodiment 65 wherein E is phenyl optionally substituted with up to three substituents independently selected from R 21 , R 22 and R 23 .
  • Embodiment 67 A pharmaceutical composition according to embodiment 66 wherein CGr is
  • Embodiment 68 A pharmaceutical composition according to any one of the embodiments 60 to 67 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 69 A pharmaceutical composition according to embodiment Error! Reference source not found, wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 70 A pharmaceutical composition according to embodiment 69 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 71 A pharmaceutical composition according to embodiment 70 wherein R 21 , R 22 and R 23 are independently selected from
  • Embodiment 72 A pharmaceutical composition according to embodiment 71 wherein R 2 ⁇ R 22 and R 23 are independently selected from
  • Embodiment 73 A pharmaceutical composition according to any one of the embodiments 60 to 72 wherein R 19 is hydrogen or methyl.
  • Embodiment 74 A pharmaceutical composition according to embodiment 73 wherein R 1 ⁇ is hydrogen.
  • Embodiment 75 A pharmaceutical composition according to any one of the embodiments 60 to 74 wherein R 27 is Hydrogen, Ci-Ce-alkyl or aryl.
  • Embodiment 76 A pharmaceutical composition according to embodiment 75 wherein R 27 is hydrogen or C r C e -alkyl.
  • Embodiment 77 A pharmaceutical composition according to any one of the embodiments 60 to 76 wherein R 28 is hydrogen or C r C ⁇ -alkyl.
  • Embodiment 78 A pharmaceutical composition according to embodiment 60 wherein F is a valence bond.
  • Embodiment 79 A pharmaceutical composition according to embodiment 60 wherein F is Cr
  • Embodiment 80 A pharmaceutical composition according to any one of the embodiments 60 or 78 to 79 wherein G is Ci-C ⁇ -alkyl or aryl, wherein the aryl is optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • Embodiment 81 A pharmaceutical composition according to any one of the embodiments 60 or 78 to 79 wherein G is C r Ce-alkyl or ArGI 1 wherein the aryl is optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • Embodiment 82 A pharmaceutical composition according to embodiment 80 wherein G is
  • Embodiment 83 A pharmaceutical composition according to embodiment 82 wherein G is phenyl optionally substituted with up to three substituents R 24 , R 25 and R 26 .
  • Embodiment 84 A pharmaceutical composition according to any one of the embodiments 60 to 83 wherein R 24 , R 25 and R 28 are independently selected from
  • Embodiment 85 A pharmaceutical composition according to embodiment 84 wherein R 24 , R 25 and R 28 are independently selected from
  • Embodiment 86 A pharmaceutical composition according to embodiment 85 wherein R 24 , R 25 and R 26 are independently selected from
  • Embodiment 87 A pharmaceutical composition according to embodiment 86 wherein R 2 ⁇ R 22 and R 23 are independently selected from
  • Embodiment 88 A pharmaceutical composition according to embodiment 87 wherein R 21 , R 22 and R 23 are independently selected from
  • ArGI-d-Cs-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se ⁇ lected from R 30 .
  • Embodiment 90 A pharmaceutical composition according to any one of the embodiments 60 or 78 to 89 wherein R 20 is hydrogen or methyl.
  • Embodiment 91 A pharmaceutical composition according to embodiment 90 wherein R 20 is hydrogen/**
  • Embodiment 92 A pharmaceutical composition according to any one of the embodiments 60 or 78 to 91 wherein R 27 is hydrogen, Ci-C 6 -alkyl or aryi.
  • Embodiment 93 A pharmaceutical composition according to embodiment 92 wherein R 27 is hydrogen or Ci-C ⁇ -alkyl or ArG1.
  • Embodiment 94 A pharmaceutical composition according to embodiment 93 wherein R 27 is hydrogen or C r C ⁇ -alkyl.
  • Embodiment 95 A pharmaceutical composition according to any one of the embodiments 60 or 78 to 93 wherein R 28 is hydrogen or d-Ce-alkyl.
  • Embodiment 96 A pharmaceutical composition according to embodiment 60 wherein R 17 and
  • R 18 are independently selected from
  • Embodiment 97 A pharmaceutical composition according to embodiment 96 wherein R 17 and R 18 are independently selected from
  • Embodiment 98 A pharmaceutical composition according to embodiment 97 wherein R 17 and R 1 ⁇ are independently selected from
  • Embodiment 99 A pharmaceutical composition according to embodiment 98 wherein R 17 and R 18 are independently selected from
  • Ci-C ⁇ -alkyl optionally substituted with one or more substituents independently se ⁇ lected from R 29 -phenyl, phenyloxy, phenyl-d-C ⁇ -aikoxy, phenyl-Ci-C ⁇ -alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se ⁇ lected from R 30 .
  • Embodiment 101 A pharmaceutical composition according to any one of the embodiments.
  • R 27 is hydrogen or Ci-C 6 -alkyl.
  • Embodiment 102 A pharmaceutical composition according to embodiment 101 wherein R 27 is hydrogen, methyl or ethyl.
  • Embodiment 103 A pharmaceutical composition according to any one of the embodiments
  • R 2 ⁇ is hydrogen or C r C 6 -alkyl.
  • Embodiment 104 A pharmaceutical composition according to embodiment 103 wherein R 28 is hydrogen, methyl or ethyl.
  • Embodiment 105 A pharmaceutical composition according to any one of the embodiments
  • R 72 is -OH or phenyl.
  • Embodiment 106 A pharmaceutical composition according to embodiment 60 wherein CGr is
  • Embodiment 107 A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is of the form H-I-J-
  • Z 1 is S(O) 2 or CH 2
  • Z 2 is -NH-, -O-or -S-
  • ⁇ is 1 or 2
  • R 31 is independently selected from hydrogen, halogen, S CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 , -SCF 3 , -NO 2 .
  • -OR 35 , -C(O)R 35 . -NR 35 R 36 , -SR 35 .
  • R 32 and R 33 are independently selected from hydrogen, Ci-C ⁇ -alkyl or CrC 6 -alkanoyl,
  • R 34 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR 35 R 38 ,
  • R 35 and R 3 ⁇ are independently selected from hydrogen, d-C ⁇ -alkyl. aryl-Ci-C 6 -alkyl or aryl, or R 35 and R 3 ⁇ when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 37 is independently selected from halogen, -C(O)OR 33 , -C(O)H, -CN, -CF 3 , -OCF 3 , -NO 2 . - OR 35 . -NR 35 R 36 , d-C ⁇ -alkyl or C r Ce-alkanoyl,
  • Embodiment 108 A pharmaceutical composition according to embodiment 107 wherein CGr is of the form H-I-J, wherein H is
  • phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R 31 ,
  • I is selected from *a valence bond
  • n 1 or 2
  • J is • d-C ⁇ -alkyl, CjrC ⁇ -alkenyl or Cz-C ⁇ -alkynyl, which may each optionally be substituted with one or more substituents selected from R 34 ,
  • R 31 is independently selected from hydrogen, halogen, -CN, -CH 2 CN 1 -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 , -OCF 2 CHF 2 , -S(O) 2 CF 3 , -SCF 3 , -NO 2 , -OR 35 , -C(O)R 35 , -NR 35 R 36 , -SR 35 , -NR 35 S(O) 2 R 38 , -S(O) 2 NR 35 R 38 , -S(O)NR 35 R 36 .
  • R 32 and R 33 are independently selected from hydrogen. d-Cg-alkyl or CrC ⁇ -alkanoyl,
  • R 34 Js independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR 35 R 38 ,
  • R 35 and R 3 ⁇ are independently selected from hydrogen, Ci-C ⁇ -alkyl, aryl-Ci-C ⁇ -alkyl or aryl. or R 35 and R 36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 37 is independently selected from halogen, -C(O)OR 35 , -C(O)H, -CN, -CF 3 , -OCF 3 , -NO 2 , - OR 35 , -NR 35 R 38 , d-Ce-alkyl or Ci-C B -alkanoyl,
  • Embodiment 109 A pharmaceutical composition according to any one of the embodiments 107 or 108 wherein H is
  • Embodiment 110 A pharmaceutical composition according to embodiment 109 wherein H is
  • Embodiment 111 A pharmaceutical composition according to embodiment 109 wherein H is
  • Embodiment 112 A pharmaceutical composition according to any one of the embodiments 107 to 111wherein I is a valence bond, -CH 2 N(R 32 )-, or SO 2 N(R 33 )-.
  • Embodiment 113 A pharmaceutical composition according to embodiment 112 wherein I is a valence bond.
  • Embodiment 114 A pharmaceutical composition according to any one of the embodiments
  • Ci-Ce-alkyl Ca-Ce-alkenyl or C 2 -C ⁇ -alkynyl, which may optionally be substituted with one or more substituents selected from halogen.
  • -CN, -CF 3 , -OCF 3 , -OR 35 , and -NR 35 R 36
  • Embodiment 115 A pharmaceutical composition according to embodiment 114 wherein J is
  • Embodiment 116 A pharmaceutical composition according to embodiment 114 wherein J is
  • Embodiment 117 A pharmaceutical composition according to embodiment 116 wherein J is • hydrogen,
  • Embodiment 118 A pharmaceutical composition according to embodiment 117 wherein J is hydrogen.
  • Embodiment 119 A pharmaceutical composition according to any one of the embodiments
  • R 32 and R 33 are independently selected from hydrogen or d-C E -alkyl.
  • Embodiment 120 A pharmaceutical composition according to any one of the embodiments.
  • R 34 is hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -SCF 3 , -NO 2 , -OR 35 .
  • Embodiment 121 A pharmaceutical composition according to embodiment 120 wherein R 34 is hydrogen, halogen, -CF 3 . -NO 2 , -OR 35 , -NR 35 R 36 , -SR 35 , -NR 35 C(O)R 38 , Or-C(O)OR 35 .
  • Embodiment 122 A pharmaceutical composition according to embodiment 121 wherein R 34 is hydrogen, halogen, -CF 3 , -NO 2 , -OR 35 , -NR 35 R 38 . or -NR 35 C(O)R 36 .
  • Embodiment 123 A pharmaceutical composition according to embodiment 122 wherein R 34 is hydrogen, halogen, or -OR 35 .
  • Embodiment 124 A pharmaceutical composition according to any one of the embodiments
  • R 35 and R 3 ⁇ are independently selected from hydrogen, d-C ⁇ -alkyl, or aryl.
  • Embodiment 125 A pharmaceutical composition according to embodiment 124 wherein R 35 and R 39 are independently selected from hydrogen or Ci-C ⁇ -alkyl.
  • Embodiment 126 A pharmaceutical composition according to any one of the embodiments
  • R 37 is halogen, -C(O)OR 35 . -CN, -CF 3 , -OR 33 , -NR 35 R 36 , d-C ⁇ -alkyl or C 1 - C 6 -alkanoyl.
  • Embodiment 127 A pharmaceutical composition according to embodiment 126 wherein R 37 is halogen, -C(O)OR 35 , -OR 35 , -NR 35 R 36 , CrC ⁇ -alkyl or C r C ⁇ -alkanoyl.
  • Embodiment 128 A pharmaceutical composition according to embodiment 127 wherein R 37 is halogen, -C(O)OR 35 or -OR 35 .
  • Embodiment 129. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
  • U is a valence bond, d-C ⁇ -alkenylene, -d-C 6 -alkyl-O- or Ci-C ⁇ -alkylene wherein any C 1 - Ce-alkyl moiety is optionally substituted with Ci-Ce-alkyl, R 38 is CrC ⁇ -alky], aryl, wherein the alky] or aryl moieties are optionally substituted with one or more substituents independently selected from R 3 ⁇ ,
  • R 39 is independently selected from halogen, cyano, ⁇ itro, amino,
  • M is a valence bond, afylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R 40 ,
  • R 40 is selected from •hydrogen, halogen, -CN, -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 . -OCHF 2 , -OCH 2 CF 3 .
  • R 41 and R 42 are independently selected from hydrogen, -OH 1 Ci-C ⁇ -alkyl, CrC 6 -alkenyl, aryl- Ci-C ⁇ -afkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R 45 , and the aryl moieties may optionally be substi ⁇ tuted with one or more substituents independently selected from R 46 ; R 41 and R 42 when at ⁇ tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R 43 is independently selected from halogen, -CN. -CF 3 , -OCF 3 , -OR 41 , and -NR 41 R 42
  • R 44 is independently selected from halogen, -C(O)OR 41 , -CH 2 C(O)OR 41 , -CH 2 OR 41 , -CN, - CF 3 , -OCF 3 , -NO 2 , -OR 41 , -NR 41 R 42 and C r C 6 -alkyl,
  • R 45 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -O-d-C ⁇ -alkyl, -C(O)-O-C 1 -
  • R 46 is independently selected from halogen, -C(O)OCi-C ⁇ -alkyl, -COOH, -CN, -CF 3 , -OCF 3 , -
  • Q is a valence bond, d-C ⁇ -alkyle ⁇ e. -d-Ce-alkyl-O-, -d-C ⁇ -alkyl-NH-, -NH-C-C ⁇ -alkyl,
  • R 47 and R 4 ⁇ are independently selected from hydrogen, d-C ⁇ -alkyl, aryl optionally substituted with one or more R 49 .
  • R 49 is independently selected from halogen and -COOH
  • Ci-C ⁇ -alkyl C 2 -C 6 -alkenyl , C 2 -C ⁇ -alkynyl, d-C ⁇ -alkyloxy-carbonyl, wherein the alkyl, alkenyl and alkynyl moieties are optionally substituted with one or more substituents independently selected from R 50 ,
  • any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R 50 ,
  • Embodiment R 50 * and R 50B are independently selected from -C(O)OC r C ⁇ -alkyl, -COOH, -C 1 - C ⁇ -alkyl-CtOOC-Ce-alkyl, -C r C ⁇ -aikyUCOOH. or C,-C ⁇ -alkyl,
  • R 51 and R 52 are independently selected from hydrogen and d-C ⁇ -alkyl
  • R 53 Is independently selected from C-rC e -alkyl, d-C ⁇ -alkoxy, -d-C ⁇ -alkyl-COOH, -C 2 -
  • a pharmaceutical composition according to embodiment 130 wherein K is a valence bond, d-C B -alkylene, -NH-C( O)-U-, -Ci-C ⁇ -alkyl-S-, or -d-C 6 -alkyl-O, wherein any d-C ⁇ -alkyl moiety is optionally substituted with R 38 .
  • Embodiment 133 A pharmaceutical composition according to embodiment 132 wherein K is a valence bond or d-C a -alkylene, wherein any d-C ⁇ -alkyl moiety is optionally substituted with R 38 .
  • Embodiment 135. A pharmaceutical composition according to embodiment 133 wherein K is a valence bond.
  • Embodiment 136 A pharmaceutical composition according to any one of the embodiments
  • Embodiment 137 A pharmaceutical composition according to embodiment 136 wherein U is a valence bond.
  • Embodiment 138 A pharmaceutical composition according to any one of the embodiments
  • Embodiment 139 A pharmaceutical composition according to embodiment 138 wherein M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • Embodiment 139 A pharmaceutical composition according to embodiment 138 wherein M is
  • Embodiment 140 A pharmaceutical composition according to embodiment 139 wherein M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R 40 .
  • Embodiment 141 A pharmaceutical composition according to embodiment 140 wherein M is
  • Embodiment 143 A pharmaceutical composition according to embodiment 141 wherein M is indolylene optionally substituted with one or more substituents independently selected from R 40 .
  • Embodiment 144 A pharmaceutical composition according to embodiment 143 wherein M is
  • Embodiment 146 A pharmaceutical composition according to embodiment 145 wherein M is
  • Embodiment 147 A pharmaceutical composition according to any one of the embodiments 129 to 146 wherein R 40 is selected from -hydrogen, halogen, -CN, -CF 3 , -OCF 3 , -NO 2 , -OR 41 , -NR 41 R 42 , -SR 41 , -S(O) 2 R 41 .
  • CrC ⁇ -alkyl or C 2 -C 8 - alkenyl which may each optionally be substituted with one or more substituents independently selected from R 43 , • aryl, aryloxy, ary]-Ci-C ⁇ -alkoxy, aryl-d-C ⁇ -alkyl, aryl-C ⁇ Ce-alkenyl, heteroaryl, het- eroaryl-CrCe-alkyl, or heteroaryt-C 2 -C e -alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substitue ⁇ ts selected from R 44 .
  • Embodiment 148 A pharmaceutical composition according to embodiment 147 wherein R* 0 is selected from
  • Embodiment 149 A pharmaceutical composition according to embodiment 148 wherein R 40 is selected from
  • Embodiment 150 A pharmaceutical composition according to embodiment 149 wherein R 40 is hydrogen.
  • Embodiment 151 A pharmaceutical composition according to embodiment 149 wherein R 40 is selected from • halogen, -NO 2 , -OR 41 , -NR 41 R 42 , -C(O)OR 41 , or -NR 41 C(O)R 42 ,
  • Embodiment 152 A pharmaceutical composition according to any one of the embodiments 129 to 151 wherein R 41 and R 42 are independently selected from hydrogen, d-C ⁇ -alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
  • Embodiment 153 A pharmaceutical composition according to embodiment 152 wherein R 41 and R 42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or -COOH.
  • Embodiment 156 A pharmaceutical composition according to any one of the embodiments 129 to 155 wherein R 47 and R 48 are independently selected from hydrogen, methyl and phenyl.
  • Embodiment 157 A pharmaceutical composition according to any one of the embodiments 129 to 156 wherein T is
  • Ci-C 6 -alkyl optionally substituted with one or more substituents independently se ⁇ lected from R 50 ,
  • Embodiment 158 A pharmaceutical composition according to embodiment 157 wherein T is
  • Ci-C 6 -alkyl optionally substituted with one or more substituents independently se ⁇ lected from R 50 , »ArG1, ArG1-Ci-C e -atkyl, Het3, wherein the alkyl, aryl and heteroaryl moieties are op- tionally substituted with one or more substituents independently selected from R .
  • Embodiment 159 A pharmaceutical composition according to embodiment 158 wherein T is
  • Embodiment 160 A pharmaceutical composition according to embodiment 159 wherein T is phenyl substituted with R 50 .
  • Embodiment 163 A pharmaceutical composition according to embodiment 162 wherein R 50 is CrC ⁇ -alkyl, aryloxy, aryl-Ct-C ⁇ -alkoxy, -OR 51 , halogen, -COOH 1 -CF 3 , wherein any aryl moiety is optionally substi ⁇ tuted with one or more R 53 .
  • Embodiment 165 A pharmaceutical composition according to embodiment 164 wherein R 50 is Embodiment 166.
  • Embodiment 167 A pharmaceutical composition according to embodiment 166 wherein R 50 is methyl or ethyl.
  • Embodiment 168. A pharmaceutical composition according to any one of the embodiments 129 to 167 wherein m is 1 or 2.
  • Embodiment 169 A pharmaceutical composition accord ⁇ hg to any one of the embodiments 129 to 168 wherein R 51 is methyl.
  • Embodiment 170 A pharmaceutical composition according to any one of the embodiments 129 to 169 wherein R 53 is Ci-C ⁇ -alkyl, Ci-C 6 -alkoxy, -OR 51 , halogen.or -CF 3 .
  • Embodiment 172 A pharmaceutical composition according to any one of the embodiments 129 to 171 wherein R MB is -C(O)OCH 3 , -C(O)OCH 2 CH 3 -COOH, -CH 2 C(O)OCH 3 , - CH 2 C(O)OCH 2 CH 3 , -CH 2 CH 2 C(O)OCH 3 , -CH 2 CH 2 C(O)OCH 2 CH 3 , -CH 2 COOH, methyl, or ethyl.
  • V is CrC 6 -alkyl, aryl, heteroaryl, aryl-C 1-8 -alkyl- or aryl-C 2- ⁇ -alkenyl-, wherein the al- kyl or alkenyl is optionally substituted with one or more substituents independently selected from R 54 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 5S ,
  • R 54 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2
  • R 55 is independently selected from »hydrogen, halogen, -CN. -CH 2 CN, -CHF 2 , -CF 3 , -OCF 3 , -OCHF 2 , -OCH 2 CF 3 ,
  • alkyl groups may optionally be substituted with one or more substituents independently selected from R ao
  • the aryl groups may option ⁇ ally be substituted with one or more substituents independently selected from R 61 ;
  • R 56 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
  • R 59 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , -OR 56 , and -NR 56 R 57 ,
  • R 59 is independently selected from halogen. -C(O)OR 56 , -CH 2 C(O)OR 56 , -CH 2 OR 56 , -CN, - CF 3 , -OCF 3 , -NO 2 , -OR 56 , -NR 56 R 57 and C r C ⁇ -alkyl,
  • R 62 is CrC ⁇ -alkyl, aryl optionally substituted with one or more substituents independently se ⁇ lected from halogen, or heteroaryl optionally substituted with one or more inde ⁇ pendently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
  • Embodiment 174 A pharmaceutical composition according to embodiment 173 wherein V is aryl, heteroaryl, or aryl-C 1-8 -alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected R 54 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 178 A pharmaceutical composition according to embodiment 177 wherein V is aryl optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 179 A pharmaceutical composition according to embodiment 178 wherein V is ArG1 optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 180 A pharmaceutical composition according to embodiment 179 wherein V is phenyl, naphthyl or anthranyl optionally substituted with one or more substituents independ ⁇ ently selected from R 55 .
  • Embodiment 181 A pharmaceutical composition according to embodiment 180 wherein V is phenyl optionally substituted with one or more substituents independently selected from R 55 .
  • Embodiment 182. A pharmaceutical composition according to any one of the embodiments 173 to 181 wherein R 55 is independently selected from
  • Ci-C e -alkyl optionally substituted with one or more substituents independently se- lected from R 58
  • Embodiment 183 A pharmaceutical composition according to embodiment 182 wherein R 55 is independently selected from
  • Ci-C ⁇ -alkyl optionally substituted with one or more substituents independently se ⁇ lected from R 58 • ArG1.
  • Embodiment 184 A pharmaceutical composition according to embodiment 183 wherein R 55 is independently selected from halogen, -OR 56 , -NR 56 R 57 , -C(O)OR 56 , -OC 1 -C 8 - alkyl-CfOJOR 56 , -NR 56 C(O)R 57 or C r C e -alkyl.
  • Embodiment 185 A pharmaceutical composition according to embodiment 184 wherein R 55 is independently selected from halogen, -OR 56 , -NR 56 R 57 , -C(O)OR 56 , -OC 1 -Ca- alkyl-C(O)OR 58 , -NR 56 C(O)R 57 , methyl or ethyl.
  • R 55 is independently selected from halogen, -OR 56 , -NR 56 R 57 , -C(O)OR 56 , -OC 1 -Ca- alkyl-C(O)OR 58 , -NR 56 C(O)R 57 , methyl or ethyl.
  • Embodiment 186 Embodiment 186.
  • a pharmaceutical composition according to any one of the embodiments 173 to 185 wherein R 58 and R 57 are independently selected from hydrogen, CF 3 , Ci-Ci2-alkyI, or -C( O)-C 1 -Ca-alkyl; R 59 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • Embodiment 187 A pharmaceutical composition according to embodiment 186 wherein R 56 and R 57 are independently selected from hydrogen or C r CiralkyL R 5 ⁇ and R 57 when at- tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • Embodiment 188 A pharmaceutical composition according to embodiment 187 wherein R 56 and R 57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R 58 and R 57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
  • Embodiment 189 A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
  • AA is Ci-C 6 -alkyl, aryl, heteroaryl, or aryl-C 2 - 6 -alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R 63 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 64 ,
  • R 63 is independently selected from halogen, -CN, -CF 3 , -OCF 3 , aryl, -COOH and -NH 2 .
  • R 64 is independently selected from
  • R ⁇ 7 is independently selected from halogen, -CN, -CF 3 , -OCF 3 . -OR 85 , and -NR 65 R 68 ,
  • R 6B is independently selected from halogen, -C(O)OR 65 , -CH 2 C(O)OR 65 , -CH 2 OR 65 , -CN, - CF 3 , -OCF 3 , -NO 2 , -OR 65 , -NR 65 R 66 and Ci-C ⁇ -alkyl,
  • R ⁇ is independently selected from C t -C ⁇ -alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more Ci-C ⁇ -alkyl,
  • R 70 is independently selected from halogen, -CN. -CF 3 , -OCF 3 , -OC r C ⁇ -alkyl, -C(O)OC 1 -C 6 - alkyl, -COOH and -NH 2 ,
  • Embodiment 190 A pharmaceutical composition according to embodiment 189 wherein AA is aryl, heteroaryl or aryl-C 1-8 -alkyl-, wherein the alkyl is optionally substituted with one or more R 63 , and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R 84 .
  • Embodiment 191. A pharmaceutical composition according to embodiment 190 wherein AA is aryl or heteroaryl optionally substituted with one or more substituents independently se ⁇ lected from R ⁇ 4 .
  • Embodiment 192 A pharmaceutical composition according to embodiment 191 wherein AA is ArG 1 or Het1 optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 193 A pharmaceutical composition according to embodiment 192 wherein AA is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 194. A pharmaceutical composition according to embodiment 193 wherein AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 64 .
  • Embodiment 195 A pharmaceutical composition according to embodiment 194 wherein AA is phenyl, naphtyl, anthryl, carbazolyl, thienyl, pyridyl, or benzodioxyl optionally substituted with one or more substituents independently selected from R 84 .
  • Embodiment 196 A pharmaceutical composition according to embodiment 195 wherein AA is phenyl or naphtyl optionally substituted with one or more substituents independently se ⁇ lected from R 64 .
  • Embodiment 197 A pharmaceutical composition according to any one of the embodiments 189 to 196 wherein R 84 is independently selected from hydrogen, halogen, -CF 3 , -OCF 3 , -OR 65 . -NR 6S R ⁇ , d-Ce-alkyl , -OC(O)R 65 , -OCi-C 6 -alkyl-C(O)OR e5 .
  • Embodiment 198 A pharmaceutical composition according to embodiment 197 wherein R 64 is independently selected from halogen, -CF 3 , -OCF 3 , -OR 65 , -NR 65 R 86 , methyl, ethyl, propyl, -OC(O)R 65 , -OCH r C(O)OR 65 . -OCHrCH 2 -C(O)OR 65 , phenoxy optionally substituted with one or more substituents independently selected from R 68 .
  • Embodiment 199 A pharmaceutical composition according to any one of the embodiments 189 to 198 wherein R 65 and R ⁇ are independently selected from hydrogen, CF 3 , Ci-Cu-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently se ⁇ lected from R 71 .
  • Embodiment 200 A pharmaceutical composition according to embodiment 199 wherein R 85 and R 88 are independently hydrogen, C r C 12 -aIkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 201 A pharmaceutical composition according to embodiment 200 wherein R 65 and R ⁇ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-d ⁇ methyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 202 A pharmaceutical composition according to embodiment 201 wherein R 65 and R ⁇ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG 1 or Het2 optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 203 A pharmaceutical composition according to embodiment 202 wherein R 65 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 204 A pharmaceutical composition according to embodiment 203 wherein R ⁇ 5 and R 66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R 71 independently; or isoxazolyl optionally substituted with one or more substituents independently selected from R 71 .
  • Embodiment 205 A pharmaceutical composition according to any one of the embodiments
  • R 71 is halogen or d-C ⁇ -alkyl.
  • Embodiment 206 A pharmaceutical composition according to embodiment 205 wherein R 71 is halogen or methyl.
  • Embodiment 207 A pharmaceutical preparation according to any one of the embodiments 1 to 205 wherein Frg consists of 0 to 5 neutral amino acids independently selected from the group consisting of GIy 1 Ala, Thr, and Ser.
  • Embodiment 208 A pharmaceutical preparation according to embodiment 207 wherein Frg consists of 0 to 5 GIy.
  • Embodiment 209 A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 0 GIy.
  • Embodiment 210 A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 1 GIy.
  • Embodiment 211. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 2 GIy.
  • Embodiment 212 A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 3 GIy.
  • Embodiment 213. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 4 GIy.
  • Embodiment 214. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 5 GIy.
  • Embodiment 215. A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein G B is of the formula B 1 -B 2 -C(O)-, B 1 -B 2 -SOz- or B 1 ⁇ -CHz- . wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 216 A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein G B is of the formula B 1 -B 2 -C(OK B 1 ⁇ -SO 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in embodiment 1.
  • Embodiment 217 A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein G B is of the formula B'-B ⁇ CfO)-, B 1 ⁇ -CH 2 - or B 1 -B 2 -NH-, wherein B 1 and B 2 are as defined in embodiment 1.

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Abstract

Novel preparations comprising ligands for the HisB10 Zn2+ sites of the R-state insulin hexamer wherein the ligand is extended by protamine that are capable of prolonging the ac­tion of insulin preparations.

Description

PHARMACEUTICAL PREPARATIONS COMPRISING INSULIN.
FIELD OF THE INVENTION
The present invention discloses insulin preparations comprising ligands for the HisB10-Zn2+ sites of the R-state insulin hexamer wherein the ligand is extended by protamine.
BACKGROUND OF THE INVENTION
Insulin AHostery. The insulin hexamer is an allosteric protein that exhibits both posi¬ tive and negative cooperativity and half-of-the-sites reactivity in ligand binding. This allosteric behaviour consists of two interrelated allosteric transitions designated LA 0 and LB 0, three in¬ ter-converting allosteric conformation states (eq. 1),
I A ( B
L o L o
T8 «→ T3R3 -M. R8 (1)
designated T61 T3R3, and R6 and two classes of allosteric ligand binding sites designated as the phenolic pockets and the His610 anion sites. These allosteric sites are associated only with insulin subunits in the R conformation. insulin Hexamer Structures and Ligand Binding. The T- to R-transition of the insulin hexamer involves transformation of the first nine residues of the B chain from an extended conformation in the T-state to an a-helical conformation in the R-state. This coil-to-heJix transition causes the N-terminal residue, PheB1, to undergo an ~ 30 A change in position. This conformational change creates hydrophobic pockets (the phenolic pockets) at the sub- unit interfaces (three in T3R3, and six in R8), and the new B-chain helices form 3-helix bun¬ dles (one in T3R3 and two in R6) with the bundle axis aligned along the hexamer three-fold symmetry axis. The His910 Zn2+ in each R3 unit is forced to change coordination geometry from octahedral to either tetrahedra! (monodentate ligands) or pentahedral (bidentate ligands). Formation of the helix bundle creates a narrow hydrophobic tunnel in each R3 unit that extends from the surface -12 A down to the HisB1° metal ion. This tunnel and the HisB1° Zn2+ ion form the anion binding site.
Hexamer Ligand Binding and Stability of Insulin Formulations. The in vivo role of the T to R transition is unknown. However, the addition of allosteric ligands (e.g. phenol and chloride ion) to insulin preparations is widely used. Hexamerization is driven by coordination of Zn2+ at the His810 sites to give T6, and the subsequent ligand-mediated transition of T6 to T3R3 and to R6 is known to greatly enhance the physical and chemical stability of the result¬ ing formulations. Ligand Binding and Long Acting Insulin Formulations. Although the conversion of T6 to T3R3 and Re improves the stability of the preparation, the rate of absorption following subcutaneous injection of a soluble hexameric preparation is not much affected by the addi¬ tion of phenol and cloride. Putative events following injection of a soluble hexameric preparation. The small molecule ligands initially diffuse away from the protein. The affinity of the ligands for insulin may help to slow this process. On the other hand, trie affinity of Zn2+ for e.g. albumin and the large effective space available for diffusion of the lipophilic phenol will tend to speed up the separation. In about 10-15 minutes after injection, the distribution of insulin species in the subcutaneous tissue will roughly correspond to that of a zinc-free insulin preparation at the same dilution. Then, the equilibrium distribution of species at this point will determine the ob¬ served absorption rate. In this regimen, absorption rates vary between about 1 hour (for rapid-acting insulin analogues, such as AspB2β human insulin) and about 4 hours (Co3+- hexamer). Current Approaches Toward Slow Acting Insulins. The inherent limitation of the ab¬ sorption half-life to about 4 hours for a soluble human insulin hexamer necessitates further modifications to obtain the desired protraction. Traditionally, this has been achieved by the use of preparations wherein the constituent insulin is in the form of a crystalline and/or amor¬ phous precipitate. In this type of formulation, the dissolution of the precipitate in the subcuta- neous depot becomes rate-limiting for the absorption. NPH and Ultratente belong to this category of insulin preparations where crystallization/precipitation is effected by the addition of protamine and excessive zinc ion, respectively.
Most recently, a series of soluble insulin derivatives with a hydrophobic moiety cova- lently attached to the side chain of LysB29 have been synthesized. These derivatives may show prolonged action profile due to various mechanisms including albumin binding (e.g. B29-Nε-myristoyl-des(B30) human insulin), extensive protein self-association and/or sticki¬ ness (e.g. B29-Nε-(N-lithocholyl-γ-glutamyl)-des(B30) human insulin) induced by the attached hydrophobic group.
SUMMARY OF THE INVENTION The present invention provides insulin preparations comprising high-affinity ligands for the HisBi0-Zn2+ sites of the R-state insulin hexamer, zinc ions and insulin wherein the ligand is extended by protamine.
The resulting ligands with protamine extensions work to modify the time action pro¬ file of insulin formulations. These preparations may be formulated with variable insulin spe- cies over a wide range of pH from 3.0 to 8.5 and their time action profiles may be tailored by suitable adjustments of anchor affinity as well as the concentration of protamiπe-extended iigand.
The invention also provides a method of preparing ligands for the His810 Zn2+ sites of the R-state insulin hexamer comprising the steps of • Identifying a starter compound that binds to the R-state HisB10-Znz+ site
• optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids « attaching protamine
Also provided are methods of treating type 1 or type 2 diabetes comprising adminis¬ tering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation of the invention.
DEFINITIONS
The following is a detailed definition of the terms used to describe the invention: "Halogen" designates an atom selected from the group consisting of F1 Cl, Br and \. The term "C1-C6-BIkVl11 as used herein represents a saturated, branched or straight hydrocarbon group having from 1 to 6 carbon atoms. Representative examples include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, n- pentyl, isopentyl, neopentyl, fert-peπtyl, n-hexyl, isohexyl and the like.
The term "CrCe-alkylene" as used herein represents a saturated, branched or straight bivalent hydrocarbon group having from 1 to 6 carbon atoms. Representative examples in- elude, but are not limited to, methylene, 1 ,2-ethylene, 1 ,3-propylene, 1 ,2-proρylene, 1 ,4- butylene, 1 ,5-pentylene, 1 ,6-hexyleπe, and the like.
The term "C2-Ce-alkenyr as used herein represents a branched or straight hydro¬ carbon group having from 2 to 6 carbon atoms and at least one double bond. Examples of such groups include, but are not limited to. vinyl, 1-propenyl, 2-propenyl, iso-propenyl, 1,3- butadienyl, 1-butenyl, 2-butenyl, 3-butenyl, 2-methyH-propenyl, 1-pentenyl, 2-pentenyl, 3- pentenyl, 4-pentenyl, 3-methyl-2-butenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 2,4-hexadienyl, 5- hexenyl and the like.
The term "C2-C6-alkynyr as used herein represents a branched or straight hydro¬ carbon group having from 2 to 6 carbon atoms and at least one triple bond. Examples of such groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2- butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-peπtynyl, 1-hexynyl, 2-hexynyl, 3- hexynyl. 4-hexynyl, 5-hexynyl, 2,4-hexadiynyl and the like.
The term "d-Ce-alkoxy" as used herein refers to the radical -O-d-CB-alkylt wherein CrC6-alkyl is as defined above. Representative examples are methoxy, ethoxy, n-propoxy, iso- propoxy, butoxy, sec-butoxy, fert-butoxy, pentoxy, isopentoxy, hexoxy, isohexoxy and the like. The term "C3-C8-cycloalkyr as used herein represents a saturated, carbocyclic group having from 3 to 8 carbon atoms. Representative examples are cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl and the like.
The term "d-β-cycloalkenyF as used herein represents a non-aromatic, carbocyclic group having from 4 to 8 carbon atoms containing one or two double bonds. Representative examples are 1-cyclopenteπyl, 2-cyclopentenyl, 3-cyclopentenyl, 1-cycIohexenyl, 2-cyclo- hexenyl, 3-cyclohexenyl, 2-cycloheptenyl, 3-cycloheptenyl, 2-cyclooctenyl, 1 ,4-cycioocta- dienyl and the like.
The term "heterocydyr as used herein represents a non-aromatic 3 to 10 membered ring containing one or more heteroatoms selected from nitrogen, oxygen and sulphur and op¬ tionally containing one or two double bonds. Representative examples are pyrrolidinyl, piperi- dyl, piperazinyl, morpholinyl, thiomorpholinyl, aziridinyl, tetrahydrofuranyl and the like.
The term "aryl" as used herein is intended to include carbocyclic, aromatic ring sys¬ tems such as 6 membered monocyclic and 9 to 14 membered bi- and tricyclic, carbocyclic, aromatic ring systems. Representative examples are phenyl, biphenylyl, naphthyl, anthra- cenyl, phenanthrenyl, fluorenyl, indenyl, a∑υlenyl and the like. Any! is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated derivatives are 1,2,3,4-tetrahydronaphthyl, 1,4- dihydronaphthyl and the like. The term "arylene" as used herein is intended to include divalent, carbocyclic, aro-
"'matic ring systems such as 6 membered monocyclic and 9 to 14 raembered bi- and tricyclic, divalent, carbocyclic, aromatic ring systems. Representative examples are phenylene, bi- phenylylene, naphthylene, anthracenylene, phenanthrenylene, fluorenylene, indenylene, az- ulenylene and the like. Arylene is also intended to include the partially hydrogenated deriva- fives of the ring systems enumerated above. Non-limiting examples of such partially hydro¬ genated derivatives are 1 ,2,3,4-tetrahydronaphthyleπe, 1,4-dihydronaphthylene and the like. The term "aryloxy" as used herein denotes a group -O-aryl, wherein aryl is as defined above.
The term "aroyl" as used herein denotes a group -C(O)-aryl, wherein aryl is as defined above.
The term "heteroaryl" as used herein is intended to include aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxygen and sul¬ phur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxy- gen and sulphur. Representative examples are furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyra- zolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1 ,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1 ,2,4-triazinyl, 1 ,3,5- triazinyl, 1 ,2,3-oxadiazolyl. 1 ,2,4-oxadiazolyl, 1 ,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1 ,2,4-thiadiazolyl, 1,2,5-thiadiazolyI, 1,3,4-thiadiazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryi, benzothienyl, indazolyl, benzimidazolyl, benzthiazolyl, beπzisothiazolyl, benzoxa- zolyl, benzisoxazolyl, purinyl, quinazolinyl, quinolizinyl, quinolinyl, isoquiπoltnyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyi, azepinyl, diazepinyl, acridinyl, thiazolidinyl, 2-thiooxo- thiazolidinyl and the like. Heteroaryl is also intended to include the partially hydrogenated de¬ rivatives of the ring systems enumerated above. Non-limiting examples of such partially hy¬ drogenated derivatives are 2,3-dihydroben∑ofuranyl, pyrrolinyl, pyrazoltnyl, indolinyl, oxazolid- tnyl, oxazoϋnyl, oxazepinyl and the like.
The term "heteroarylene" as used herein is intended to include divalent, aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitrogen, oxy¬ gen and sulphur such as 5 to 7 membered monocyclic and 8 to 14 membered bi- and tricyclic aromatic, heterocyclic ring systems containing one or more heteroatoms selected from nitro- gen, oxygen and sulphur. Representative examples are furylene, thienylene, pyrrolylene, oxazolylene, thiazolylene, imidazolylene, isoxazolylene, isothiazolylene, 1,2,3-triazolylene, 1,2,4-triazolylene, pyranylene, pyridytene, pyridazinylene, pyrimidinylene, pyrazinylene, 1,2,3-triazinylene, 1 ,2,4-triazinylene, 1,3,5- triazinylene, 1 ,2,3-oxadiazolylene, 1,2,4-oxa- diazolylene, 1,2,5-oxadiazolylene, i,3,4-oxadiazolylene, 1,2,3-thiadiazαlylene, 1,2,4-thia- diazolylene, 1,2,5-thiadiazotylene, 1,3,4-thiadiazolyleπe, tetrazolylene, thiadiazinylene, indo- lylene, isoindolylene, benzofurylene, benzothienylene, indazolylene, benzimidazolylene, benzthiazolylene, benzisothiazolylene, benzoxazolylene, benzisoxazolylene, purinylene, quinazolinylene, quinolizinylene, quinolinylene, isoquinolinylene, quinoxalinylene, naphthy- ridinylene, pteridinylene. carbazolylene, azepinylene, diazepinylene, acridinylene and the like. Heteroaryl is also intended to include the partially hydrogenated derivatives of the ring systems enumerated above. Non-limiting examples of such partially hydrogenated deriva¬ tives are 2,3-dihydrobenzofuranylene, pyrrolinylene, pyrazolinylene, indolinylene, oxazolid- inylene, oxazolinylene, oxazepinylene and the like.
The term "ArGI" as used herein is intended to include an aryl or arylene radical as ap- plicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, anthracenyl, phenaπthrenyl, fluorenyl, indenyl, and azulenyl as well as the coπτesponding divalent radicals.
The term "ArG2" as used herein is intended to include an aryl or arylene radical as ap¬ plicable, where aryl or arylene are as defined above but limited to phenyl, biphenylyl, naphthyl, fluorenyl, and indenyl, as well as the corresponding divalent radicals. The term "Hetr as used herein is intended to include a heteroaryl or heteroarytene radical as applicable, where heteroaryl or heteroarytene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1 ,2,3-triazinyi, 1 ,2,4-triazinyl, 1 ,3,5- triazinyl, 1 ,2.3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1 ,2,5-oxa- diazolyl, 1,3,4-oxadiazolyl, 1,2,3-thiadiazolyl, 1 ,2,4-thiadiazolyl, 1 ,2,5-thiadiazolyl, 1,3,4-thia- diazolyl, tetrazolyt, thiadiazinyl, tndolyl, isoindolyl, benzofuryl, benzothienyl, indazolyl, ben- zimϊdazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyt, benzisoxazolyl, purinyl, quinazolinyl, quinolizinyl, quinolinyl, isoquinolinyl, quinoxalinyl, naphthyridiπyl, pteridinyl. carbazolyl, a∑epinyl, diazepinyl. acridinyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrrespond- ing divalent radicals.
The term Ηet2" as used herein is intended to include a heteroaryl or heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl, 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1 ,2,4-triazolyl, pyranyl. pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, 1,2,3-triazinyl, 1 ,2,4-triazinyl. 1,3,5- triazinyl, 1,2,3-oxadiazolyl, 1 ,2,4-oxadiazolyl, 1,2,5-oxa- diazolyl, 1,3,4-oxadiazolyt, 1,2,3-thiadiazolyl, 1 ,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thia- diazolyl, tetrazolyl, thiadiazinyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, benzoxazolyi, benzisoxazolyl, quinolinyl. isoquinolinyl, qui- noxalinyl, carbazolyl, thiazolidinyl, 2-thiooxothiazolidinyl, as well as the corrresponding diva¬ lent radicals. rThe term "Het3" as used herein is intended to include a heteroaryl or Heteroarylene radical as applicable, where heteroaryl or heteroarylene are as defined above but limited to furyl, thienyl, pyrrolyl, pyrazolyl. 3-oxopyrazolyl, oxazolyl, thiazolyl, imidazolyl, isoxazolyl, isothiazolyl, 1,2,3-triazolyl, 1 ,2,4-triazolyl, pyridyl, tetrazolyl, indolyl, isoindolyl, benzofuryl, benzothienyl, benzimidazolyl, benzthiazolyl, benzisothiazolyl, beπzoxazotyl, benzisoxazolyl, quinolyl, isoquinolyl, quinoxalinyl, carbazolyl, thiazolidinyl, 2-thϊooxothiazolidinyl, as well as the corrresponding divalent radicals.
"Aryl-CrCe-alkyl". "heteroaryl-CrC6-alkyl", "aryl-Cϊ-Cβ-alkenyP etc. is intended to mean Ci-C8-alkyl or C2-Cβ-alkenyl as defined above, substituted by an aryl or heteroaryl as defined above, for example:
Figure imgf000007_0001
The term "optionally substituted" as used herein means that the groups in question are either unsubstituted or substituted with one or more of the substituents specified. When the groups in question are substituted with more than one substituent the substituents may be the same or different. Certain of the above defined terms may occur more than once in the structural formulae, and upon such occurrence each term shall be defined independently of the other. Furthermore, when using the terms "independently are" and "independently selected from" it should be understood that the groups in question may be the same or different.
The terms "treatment" and "treating" as used herein means the management and care of a patient for the purpose of combating a disease, disorder or condition. The term is intended to include the delaying of the progression of the disease, disorder or condition, the alleviation or relief of symptoms and complications, and/or the cure or elimination of the dis¬ ease, disorder or condition. The patient to be treated is preferably a mammal, in particular a human being. The term "fragment" as used herein is intended to mean a bivalent chemical group.
The term "neutral amino acid" as used herein is intended to mean any natural (cod- able) and non-natural amino acid, including α- or β-aminocarboxylic acids, including D- isomers of these (when applicable) without charges at physiologically relevant pH in the side chain, such as glycine, alanine, β-alanine, valine, leucine, isoleucine, phenylalanine, tyrosine, aspargine, glutamine, cysteine, methionine, 3-aminobenzoic acid, 4-aminobenzoic acid or the like.
The term "positively charged group" as used herein is intended to mean any phar¬ maceutically acceptable group that contains a positive charge at physiologically relevant pH, such as amino (primary, secondary and tertiary), ammonium and guanidino groups. The term "σ amino acid" as used herein is intended to mean mean any natural (cod- able) and non-natural α-aminocarboxylic acid, including D-isomers of these.
The term "β amino acid" as used herein is intended to mean any β-aminocarboxylic add, such as β-alanine, isoserine or the like.
The term "desB30" as used herein is intended to mean meant a natural insulin B chain or an analogue thereof lacking the B30 amino acid residue.
The amino acid residues are indicated in the three letter amino acid code or the one letter amino code.
The terms "BI", "A1" and the like as used herein is intended to mean the amino acid residue in position 1 in the B chain of insulin or analogue thereof (counted from the N- terminal end) and the amino acid residue in position 1 in the A chain of insulin or analogue thereof (counted from the N-terminal end), respectively. When in the specification or claims mention is made of groups of compounds such as carboxylates, dithiocarboxylates, pheπolates, thiophenolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano-1,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, naphthoic acids and salicylic acids, these groups of compounds are in¬ tended to include also derivatives of the compounds from which the groups take their name. The term "insulin" as used herein refers to all variants of insulin including human in¬ sulin, an analogue thereof, a derivative thereof and combinations of any of these, acid- stabilised insulin, fast/rapid acting insulin and long/stow/basal acting insulin. The term "human insulin" as used herein refers to naturally produced insulin or re- combinantly produced insulin. Recombinant human insulin may be produced in any suitable host cell, for example the host cells may be bacterial, fungal (including yeast), insect, animal or plant cells.
The term "insulin analogue" as used herein is meant human insulin in which at least one amino acid has been deleted and/or replaced by another amino acid including non- codeable amino acids, or human insulin comprising additional amino acids, i.e. more than 51 amino acids, such that the resulting analogue possesses insulin activity.
The term "insulin derivative" as used herein refers to human insulin or an analogue thereof which has been chemically modified, i.e. at least one organic substitueπt is bound to one or more of the amino acids, e.g. by introducing a side chain in one or more positions of the insulin backbone.or by oxidizing or reducing groups of the amino acid residues in theJnsulin or by converting a free carboxylic group to an ester group or acylating a free amino group or a hydroxy group.
The term "acid-stabilised insulin" as used herein refers to an insulin analog that does not deamidate or dimerize at pH values below 7. Specifically, the analog cannot have Asn or Asp as a C-terminal residue.
By "fast/rapid acting insulin" as used herein is meant any insulin having an onset of action after injection or any other form of administration faster or equal to that of soluble and neutral formulations of human insulin. The term "long/slow/basal acting insulin" as used herein is intended to include insu¬ lin compounds such as protamine insulin, zinc insulin, protamine zinc insulin.
The term "phenolic compound" or similar expressions as used herein refers to a chemical compound in which a hydroxyl group is bound directly to a benzene or substituted benzene ring. Examples of such compounds include, but are not limited to, phenol, o-cresol, m-cresol and p-cresol. "Protamine" as used herein refers to the generic name of a group of strongly basic proteins present in sperm cell nucleic in saltlike combination with nucleic acids. Commercially available protamines can be isolated from mature fish sperm and are usually obtained as the sulphate. The peptide composition of a specific protamine may vary depending of which fam- ily, genera or species of fish it is obtained from. Protamine usually contains four major com¬ ponents, i.e. single-chain peptides containing about 30-32 residues of which about 21-22 are arginines. The N-ternimal is praline for each of the four main components, and since no other amino groups are present in the sequence, chemical modification of protamine is expected to be homogenoues in this context. Normally, protamines to be used together with insulin are obtained from e.g. salmon (salmine), rainbow trout (iridine), herring (clupeine), sturgeon (sturine) or Spanish mackerel (thynnine). Protamine also refers to preparations comprising salts of the proteins.
When an insulin derivative according to the invention is stated to be "soluble at physiological pH values" it means that the insulin derivative can be used for preparing injectable insulin compositions that are fully dissolved at physiological pH values. Such favourable solubil¬ ity may either be due to the inherent properties of the insulin derivative alone or a result of a fa¬ vourable interaction between the insulin derivative and one or more ingredients contained in the vehicle.
The term "physiologically relevant pH" as used herein is intended to mean a pH of about 7.1 to 7.9.
Abbreviations:
4H3N 4-hydroxy-3-nitrobenzoic acid
AcOH acetic acid
BT Benzotriazol-5-oyl DMF N,N-Dimethylformamide
DMSO Dimethylsulfoxide
DlC Diisopropylcarbodiimidθ
EDAC 1 -ethyl-S-tS'-dimethylamino-propylJcarbodiimide, hydrochloride
Fmoc 9H-Fluorene-9-ylmethoxycarbonyl G. GIy Glycine
HOAt 1 -hydroxy-7-azabenzotriazole
HOBT 1 -Hydroxybenzotriazole
L. Lys Lysine
NMP N-methyl-2-pyrralidone Pbf 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl Pmc 2,2,5,7,8-pentamethylchroman-6-sulfonyl
R. Arg Arginine
TFA Trifluoroacetic acid
Abbreviations for non-natural amino acid residues:
Figure imgf000011_0001
BRIEF DESCRIPTION OF DRAWINGS
Fig. 1; Change in plasma glucose level after subcutaneous injection of the prepara- tion: 0,6mM A21G, B28D human insulin, 0.3mM Zn2+, 3OmM phenol, 1.6% glycerol, 0.3mM 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyrylprotamine.
Fig. 2: 4H3N-assay. UV/vis spectra resulting from a titration of hexameric insulin with the compound 3-hydroxy-2-naphthoic acid in the presence of 4-hydroxy-3-nιtrobenzoic acid (4H3N). Inserted in the upper right comer is the absorbance at 444nm vs. the coπcen- tration of ligand
Fig. 3: TZD-assay. Fluorescence spectra resulting from a titration of hexameric in¬ sulin with 5-(3-methoxybenzylidene)thiazolidine-2,4-dione in the presence of 5-(4- dimethylaminobenzylidene)thiazolidine-2T4-dione (TZD). Inserted in the upper right comer is the fluorescence at 460 nm vs. the concentration of ligand
DESCRIPTION OF THE INVENTION
The present invention is based on the discovery that the His910 Zn+* ligand binding sites of the R-state insulin hexamer wherein the ligand is extended by protamine can be used to obtain an insulin preparation having prolonged action designed for flexible injection re¬ gimes including once-daily, based on insulin molecules of any kind. The basic concept underlying the present invention involves reversible attachment of a ligand to the HisB1° Zn2+ site of the R-state hexamer. A suitable ligand binds to the hexamer metal site with one end while at the other end, the anchor features a carboxylate group. The carboxylate group is covalently bound via an amide link to protamine. On this ba¬ sis, prolonged action via modification of preparation solubility may be obtained in a number of ways. However, all cases involve the same point of protein-ligand attachment and the de¬ livery of human insulin (or analogues or derivatives thereof) as the active species. Use of a acid-stabilized insulin analog allows a stable, clear solution with ligand to be formulated at slightly acidic pH. Following subcutaneous injection, the pH is gradually adjusted towards neutral. As a result the ligand binds to and precipitates insulin in the subcutaneous tissue. The release of insulin analog into the blood stream is then limited by the rate of redissolution of the precipitate. Of particular advantage is the possibility of adjusting the amount of added ligand as well as the charge and affinity of the ligand. Variation of these parameters allows adjustment of the rate of dissolution following precipitation in the subcutis and hence the pro- portion of slow and fast acting analog in the formulationon. Hence formulations covering a wide range of release rates may be prepared by this principle.
The anions currently used in insulin formulations as allosteric ligands for the R-state hexamers (notably chloride ion) bind only weakly to the HisB1° anion site. The present inven¬ tion, which is based on the discovery of suitable higher affinity ligands for these anion sites, provides ligands which are extended to modify timing via changes in hexamer solubility as outlined above.
Most ligand binding sites in proteins are highly asymmetric. Because the HisB1° Zn2+ sites reside on the three-fold symmetry axis, these sites posses a symmetry that is unusual, but not unique. Several other proteins have highly symmetric ligand binding sites. The HisB1Q Zn2* site consists of a tunnel or cavity with a triangular-shaped cross- section that extends -12 A from the surface of the hexamer down to the His810 Zn2* ion. The diameter of the tunnel varies along its length and, depending on the nature of the ligand oc¬ cupying the site, the opening can be capped over by the Asn63 and PheB1 side chains. The walls of the tunnel are made up of the side chains of the amino acid residues along one face each of the three α-helices. The side chains from each helix that make up the lining of the tunnel are PheB1, Asn83, and LeuBe. Therefore, except for the zinc ion, which is coordinated to three HisB1° residues and is positioned at the bottom of the tunnel, the site is principally hydrophobic. Depending on the ligand structure, it may be possible for substituents on the ligand to make H-bonding interactions with AsnB3and with the peptide linkage to CysB7. The present invention originates from a search for compounds with suitable binding properties by using UV-visible and fluorescence based competition assays described herein which are based on the displacement of chromophoric ligands from the R-state HisB10-Zn2+ site by the incoming ligand in question. These compounds will be referred to as "starter com¬ pounds" in the following. These assays are easily transformed into a high-throughput format capable of handling fibraries constructed around hits from the initial search of compound da¬ tabases. These starter compounds provide the starting point for the task of constructing a chemical handle that allows for attachment of the protamine group.
Thus, from the structure-activity relationship (SAR) information obtained from the binding assay(s) it will be apparent for those skilled in the art to modify the starter com- pounds in question by introduction of a chemical group that will allow for coupling to a pep¬ tide containing e.g. one or more arginine or lysine residues. These chemical groups include carboxylic acid (amide bond formation with the peptide), carbaldehyde (reductive alkylation of the peptide), sulfonyl chloride (sulphoπamide formation with the peptide) or the like.
The decision where and how to introduce this chemical group can be made in vari- ous ways. For example: From the SAR of a series of closely related starter compounds, a suitable position in the starter compound can be identified and the chemical group can be attached to this position, optionally using a spacer group, using synthesis procedures known to those skilled in the art.
Alternatively, this chemical group can be attached (optionally using a spacer group using and synthesis procedures known to those skilled in the art) to a position on the starter compound remote from the Zn2*-binding functionality.
The invention thus provides pharmaceutical preparation comprising
1. Insulin
2. Zinc ions 3. A ligand which binds reversibly to a HisB10 Zn2+ site of an R-state hexamer and wherein the ligand is extended by covalent attachment to protamine, having the following general formula (I)
CGr-Lnk-Frg-Protamine (I)
wherein:
CGr is a chemical group which reversibly binds to a HisB1° Zn2+ site of an insulin hexamer;
Lnk is a linker selected from
• a valence bond
• a chemical group GB of the formula -B1-B2-C(O)-, -B1-B2-SO;r, -B'-B^CHa-, or -B1- B2-NH-; wherein B1 is a valence bond, -O-, -S-, or -NR6-,
B2 is a valence bond, C^C^-alkylene. C2-C1B-alkenylene, CrCi8-alkynylene, arylene. heteroarylene, -Ct-Ciβ-alkyl-aryl-, -Crde-alkenyl-aryl-. -Cz-Ciβ-alkynyl-aryl-. -C(=O)-
C1-Ciβ-alkyl-C(=O)-. -C(=O)-C1-C1β-alkenyl-C(=O)-, -C(=O)-C,-C18-alkyl-O-C1-C18- alkyl-C(=O)-, -C{=0)- C1-C18-alkyl-S-C1-C1B-alkyl-C(=O)-1 -C(=O)-CrCia-alkyl-NRβ-C1- C1β-alkyl-C(=O)-, -C(=O)-aryl-C(=O)-, -C(=O)-heteroaryl-C{=O)-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF3, -OCF3, -ORΘ. or -NR8R7 and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR6, -C(O)H, OCOR6, -SO2, -CN, -CF3, -
OCF3, -NO2, -OR6, -NR6R7, CrC18-alkyl, or Ci-C18-alkanoyl; Rβand Rr are independently H, CrC4-alkyl;
Frg is a fragment consisting of O to 5 neutral α- or β-amino acids; or
a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mix¬ ture of optical isomers, including a racernic mixture, or any tautomeric forms.
The present invention also encompasses pharmaceutically acceptable salts of the present compounds. Such salts include pharmaceutically acceptable acid addition salts, pharmaceutically acceptable metal salts, ammonium and alkylated ammonium salts. Acid addition salts include salts of inorganic acids as well as organic acids. Representative exam¬ ples of suitable inorganic acids include hydrochloric, hydrobromic, hydroiodic, phosphoric, sulphuric, nitric acids and the like. Representative examples of suitable organic acids include formic, acetic, trichloroacetic, trifluoroacetic, propionic, benzoic, cinnamic, citric, fumaric, gly- colic, lactic, maleic, malic, malonic, maπdelic, picric, pyruvic, succinic, methanesulfonic, ethanesulfonic, tartaric, ascorbic, pamoic, , ethanedisulfonic, gluconic, citraconic, aspartic, stearic, palmitic, glycolic, p-aminobenzoic, glutamic, benzenesulfonic, p-toluenesulfonic acids and the like. Further examples of pharmaceutically acceptable inorganic or organic acid addi- tion salts include the pharmaceutically acceptable salts listed in J. Pharm. Sci. 1977, 66, 2, which is incorporated herein by reference. Examples of metal salts include lithium, sodium, potassium, magnesium salts and the like. Examples of ammonium and alkylated ammonium salts include ammonium, methyl-, dimethyl-, trimethyl-, ethyl-, hydroxyethyl-, diethyl-, n-butyh sec-butyi-, tert-butyh tetramethylammonium salts and the like. Also intended as pharmaceutically acceptable acid addition salts are the hydrates, which the present compounds, are able to form.
The acid addition salts may be obtained as the direct products of compound synthe¬ sis, in the alternative, the free base may be dissolved in a suitable solvent containing the ap¬ propriate acid, and the salt isolated by evaporating the solvent or otherwise separating the salt and solvent. The compounds of the present invention may form solvates with standard low mo¬ lecular weight solvents using methods well known to the person skilled in the art. Such sol¬ vates are also contemplated as being within the scope of the present invention.
In one embodiment CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiopheπolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano-1,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thia- zolidinediones, tetrazoles, 5-mercaptotetrazoles, rhodanines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids.
In another embodiment CGr is a chemical structure selected from the group consist- ing of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidin- ediones, 5-mercaptotetrazoles, or 4-cyano-1 ,2,3-triazoles.
In another embodiment CGr is
Figure imgf000015_0001
wherein X is =O, =S or =NH Y is -S-, -O- or -NH-
R1 and R4 are independently selected from hydrogen or Ci-Cθ-alkyl, R2 is hydrogen or Ci-C6-alkyl or aryl, R1 and R2 may optionally be combined to form a double bond,
R3 and R6 are independently selected from hydrogen, halogen, aryl, CrC6-alkyl, or -C(O)NR11R12.
A and B are independently selected from d-Cβ-alkylene, arylene, aryl-CrC6-alkyl-, aryl-C2- Cβ-alkenyl- or heteroarylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R6 and the arylene or heteroarylene is optionally substituted with up to four substituents R?, R8, R9, and R10.
A and R3 may be connected through one or two valence bonds, B and R5 may be connected through one or two valence bonds, Re is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2,
R7, R8, RB and R10 are independently selected from • hydrogen, halogen, -CN, -CH2CN1 -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3. -OS(O)2CF3, -SCF3. -NO2, -OR11, -NR11R12, -SR11, -NR11S(O)2R12, -S(O)2NR11R12, -S(O)NR11R12. -S(O)R11, -S(O)2R11, -OS(O)2 R11, -C(O)NR11R12, -OC(O)NR11R12, -NR11C(O)R12, -CH2C(O)NR11R12, -OC1-C6^IKyI-C(O)NR11R12, -CH2OR11, -CH2OC(O)R11, -CH2NR11R12, -OC(O)R11,
-OCrC,5-alkyl-C(O)OR11, -OCrCe-alkyl-OR1\ -SCrCg-alkyl-CfOpR11 -C2-Cβ-alkenyl-C(=O)OR11, -NR11-C(=O)-CrCβ-alkyl-C(=O)OR11,
-NR11-C(=O)-CrCs-alkenyl-C(=O)OR11 , -C(O)OR11, C(O)R11, or -C2-Cβ-alkeπyl- C(=O)R11, =0, or-C2-Cβ-alkenyl-C(=O)-NR11R12,
• Ci-Cβ-alkyl, C∑-Cβ-alkenyl or Cz-Cβ-alkynyl. each of which may optionally be substi¬ tuted with one or more substituents independently selected from R13,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-C,-Cδ-aJkoxy, aryl-CrCe-alkyl,
Figure imgf000016_0001
aroyl-Cz-Cfl-alkenyl, aryl-C2-Cβ-alkynyl, heteroaryl, heteroaryl-Cr
Ce-alkyl, heteroaryl-Cz-Cβ-alkenyl, heteroaryl-C2-Cβ-alkynyl, or C3-C6 cycloalkyl,
of which each cyclic moiety may optionally be substituted with one or more substitu¬ ents independently selected from R1*,
R11 and R12 are independently selected from hydrogen, OH, d-Cj-o-alkyl, aryl-Ci-Cfl-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R13 is independently selected from halogen. -CN, -CF3, -OCF3, -OR11, -C(O)OR11 , -NR11R12, and -C(O)NR11R12,
R1" is independently selected from halogen, -C(O)OR11, -CH2C(O)OR11, -CH2OR11, -CN, - CF3. -OCF3, -NO2, -OR11. -NR11R12, S(O)2R11, aryl and d-Cβ-aikyl,
R15 is independently selected from halogen, -CN, -CF3, -OCF3, -Od-Cβ-alkyl, -C(O)OC1-C6- alkyl, -COOH and -NH2, Rlβ is independently selected from halogen, -C(O)OC1-Ce-SlKyI, -COOH, -CN, -CF3. -OCF3, - NO2, -OH, -OCi-Cfpalkyl, -NH2, C(=0) or Ci-Cβ-alkyl, or any enantiomer, diastereomer, in¬ cluding a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically accept- able acid or base.
In another embodiment X is =O or =S.
In another embodiment X is =O.
In another embodiment X is =S. In another embodiment Y is -O- or -S-.
In another embodiment Y is -O-.
In another embodiment wherein Y is -S-.
In another embodiment Crg is arylene optionally substituted with up to four substituents, R7,
R8, R9, and R10 which may be the same or different. In another embodiment A is selected from ArG1 optionally substituted with up to four sub¬ stituents, R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is phenylene or naphtylene optionally substituted with up to four substituents, R7, R8, Rθ, and R10 which may be the same or different.
In another embodiment A is
Figure imgf000017_0001
In another embodiment A is phenylene.
In another embodiment A is heteroarylene optionally substituted with up to four substituents,
R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is selected from Het1 optionally substituted with up to four sirb- stituents, R7, Rβ, R9, and R10 which may be the same or different.
In another embodiment A is selected from Het2 optionally substituted with up to four sub¬ stituents, R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is selected from Het3 optionally substituted with up to four sub¬ stituents, R7, R8, R9, and R10 which may be the same or different. In another embodiment A is selected from the group consisting of indolylene, benzofu- ranylidene, quinolylene, furylene, thienylene, or pyrrolylene, wherein each heteroaryl may optionally substituted with up to four substitueπts, R7, R8, R9, and R10 which may be the same or different.
In another embodiment A is beπzofuranylene optionally substituted with up to four substitu- ents R7, RB, R9, and R10 which may be the same or different. In another embodiment A is
Figure imgf000018_0001
In another embodiment A is carbazolylidene optionally substituted with up to four substitu- ents R7, R8, R9, and R10 which may be the same or different. In another embodiment A is
Figure imgf000018_0002
In another embodiment A is quinolylidene optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. In another embodiment A is
Figure imgf000018_0003
In another embodiment A is indolylene optionally substituted with up to four substituents R7, R8, Rθ, and R10 which may be the same or different. In another embodiment A is
Figure imgf000018_0004
In another embodiment R1 is hydrogen. In another embodiment R2 is hydrogen.
In another embodiment R1 and R2 are combined to form a double bond. In another embodiment R3 is d-Cβ-alkyl, halogen, or C(O)NR18R17. In another embodiment R3 is CrC6-alkyl or C(O)NR16R17. In another embodiment R3 is methyl.
In another embodiment B is phenylene optionally substituted with up to four substituents, R7, RB, R9, and R10 which may be the same or different. In another embodiment R4 is hydrogen. In another embodiment R5 is hydrogen. In another embodiment R6 is aryl. In another embodiment Rδ is phenyl. In another embodiment R7, R8, R9 and R10 are independently selected from
•hydrogen, halogen. -NO2, -OR11. -NR11R12. -SR11, -NR11S(O)2R12, -S(O)2NR11R12, -S(O)NR11R12, -S(O)R11, -S(O)2R11. -OS(O)2 R11, -NR11C(O)R12, -CH2OR11. - CH2OC(O)R11, -CH2NR1V2, -OC(O)R11, -OC1-Ce^lKyI-C(O)OR11, -OC1-C6- alkyl-C(O)NR11R12. -Od-Cβ-alkyl-OR11, -Sd-Cβ-alkyl-C(O)OR11, -QrCβ-alkenyl-
C(=O)OR11, -C(O)OR11, or -Cz-Cβ-alkenyl-C(=O)R11.
• d-C6-alkyl, C2-Cθ-alkenyl or C2-Cβ-alkynyl, which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, arylsulfanyl, aryl-d-Cβ-alkoxy, aryl-Ci-Cβ-alkyl, aryl-C2- Cβ-alkenyl, aroyl-CrCβ-alkenyl, aryl-C∑-Ce-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14
In another embodiment R7, Rβ, R8 and R10 are independently selected from
• hydrogen, halogen, -NO2, -OR11. -NR11R12, -SR11, -S(O)2R11. -OS(O)2 R". - CH2OC(O)R11, -OC(O)R11. -OC,-Ce-alkyl-C(O)OR11, -Od-Cβ-alkyl-OR11. -SC1-C6- alkyl-CtOJOR11, -C(O)OR11, or -C2-C6-alkenyl-C(=O)R11,
• CrCβ-alkyl or CrC6-alkenyl which may each optionally be substituted with one or more substituents independently selected from R13
*aryl. aryloxy, aroyl, aryl-Ci-C8-alkoxy, aryl-d-Ce-alkyl, heteroaryl, of which each of the cyclic moieties optionally may be substituted with one or more substitueπts independently selected from R"
In another embodiment R7, R8, R9 and R10 are independently selected from
•hydrogen, halogen, -NO2, -OR11. -NR11R12, -SR11, -S(O)2R11, -OS(O)2 R11. - CH2OC(O)R11. -OC(O)R11, -OCrCβ-alkyl-C(O)OR11. -Od-Cβ-alkyl-OR11. -SC1-C6- alkyl-C(O)OR11, -C(O)OR11, or-C2-C6-alkenyl-C(=O)R11,
"CrCβ-alky! or CrC6- which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, aryl-d-CB-alkoxy, aryl-C^Ce-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
In another embodiment R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11,
Figure imgf000020_0001
or -C(O)OR11,
„, •C1-Cβ-alkyl which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aryl-CrCe-alkoxy,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
In another embodiment R7, R8, R9 and R10 are independently selected from •hydrogen, halogen, -OR11, -Od-Cβ-alkyl-qojOR11, or -C(O)OR11,
• C-Ce-alkyl which may optionally be substituted with one or more substituents inde¬ pendently selected from R13 •phenyl, phenyloxy, phenyl-CrC3-alkoxy, wherein each of the cyclic moieties option¬ ally may be substituted with one or more substituents independently selected from R14.
In another embodiment R11 and R12 are independently selected from hydrogen, Ci-Ca-alkyl, aryl or aryKVCe-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from Rie; R11 and R1Z when at¬ tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds.
In another embodiment R11 and R12 are independently selected from hydrogen, CrCaralkyl, aryl or aryl-CrCβ-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R16.
In another embodiment R11 and R12 are independently selected from phenyl or phenyl-CrC6- alkyl,
In another embodiment R11 and R12 are methyl. In another embodiment R13 is independently selected from halogen, CF3, OR11 or NR11R12.
In another embodiment R13 is independently selected from halogen or OR11.
In another embodiment R13 is OR11.
In another embodiment R14 is independently selected from halogen, -C(O)OR11, -CN, -CF3, -
OR11. S(O)2R11, and CrCβ-alkyl. In another embodiment R14 is independently selected from halogen, -C(O)OR11, or -OR11.
In another embodiment R15 is independently selected from halogen, -CN, -CF3, -C(O)OC1-C6- alkyl.and -COOH.
In another embodiment R15 is independently selected from halogen or -C(O)OCi-Cβ-alkyl.
In another embodiment R1Θ is independently selected from halogen, -C(O)OCrCValkyl, -COOH, -NO2. -OCrCβ-alkyl, -NH2. C(=O) or C,-Cβ-alkyl.
In another embodiment Rtβ is independently selected from halogen,
Figure imgf000021_0001
-COOH1 -NO2, or CrC6-alkyl.
In another embodiment CGr is
Figure imgf000022_0001
wherein
R19 is hydrogen or Ci-Cθ-alkyl, R20 is hydrogen or Ci-C6-alkyl,
D and F are a valence bond or CVCβ-alkylene optionally substituted with one or more sub- stituents independently selected from R72,
R72 is independently selected from hydroxy. Ci-Cβ-alkyl, or aryl,
E is Ci-Cβ-alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is option¬ ally substituted with up to three substituents R21, R22 and R23,
G is Ci-Cβ-alkylene, arylene or heteroarylene, wherein the arylene or heteroarylene is op- tionally substituted with up to three substituents R24, R25 and R26,
R17, R18, Rz\ R22, R23, R24, R25 and R26 are independently selected from
• hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -SCF3, -NO2. -OR27, -NR27R28, -SR27, -NR27S(O)2R28, -S(O)2NR27R28, -S(O)NR27R28, -S(O)R27, -S(O)2R27, -C(O)NR27R28, -OC(O)NR27R28, -NR27C(O)R28, -NR27C(O)OR28, -CH2C(O)NR27R2*, -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28, -OC(O)R27, -OCrCβ-alkyl-C(O)0R27, -SC,-Cβ-alkyl-C(O)OR27, -C2-C6- alkenyl-C(=O)OR27, -NR27-C(=O)-CrC6-alkyl-C(=O)OR27, -NR27-C(=O)-C,-C6- aikenyl-C(=O)OR27, -C(=O)NR27-C,-C6-alkyl-C(=O)OR27,
Figure imgf000022_0002
-C(O)OR27,
> d-Cβ-alkyl, C2-Cβ-alkenyl or
Figure imgf000022_0003
which may optionally be substituted with one or more substituents independently se¬ lected from R29, •aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-CrCβ-alkyl. aryl-Car Cβ-alkenyi, aryl-Cz-Cβ-alkynyl, heteroaryl, heteroaryl-d-Cβ-alkyl, heteroaryl-C2-Cβ- alkenyl or heteroaryl-C2-Cθ-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30,
R27 and R28 are independently selected from hydrogen. CrCβ-alkyl, aryl-d-Ce-alkyl or aryl, or R27 and R2β when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R29 is independently selected from halogen, -CN, -CF3, -OCF3, -OR2r, and -NR27R28,
R30 is independently selected from halogen, -C(Q)OR27. -CM, -CF3, -OCF3. -NO2, -OR27, -NR27Rza and d-C8-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. In another embodiment D is a valence bond.
In another embodiment D is Ci-C6-alkylene optionally substituted with one or more hydroxy, d-Cβ-alkyl, or aryl.
In another embodiment E is arylene or heteroarylene, wherein the arylene or heteroarylene is optionally substituted with up to three substituents independently selected from R21, R22 and R23.
In another embodiment E is arylene optionally substituted with up to three substituents inde¬ pendently selected from R21, R22 and R23.
In another embodiment E is selected from ArG1 and optionally substituted with up to three substituents independently selected from R21, R22 and R23. In another embodiment E is pheπylene optionally substituted with up to three substituents independently selected from R21, R22 and R23. In another embodiment CGr is
Figure imgf000023_0001
In another embodiment R21, R22 and R23 are independently selected from
• hydrogen, halogen, -CHF2, -CF3. -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - NO2, -OR27, -NR27R28, -SR27. -C(O)NR27R28, -OC(O)NR27R26, -NR27C(O)R28,
-NR27C(O)OR28. -CH2C(O)NR27R28, -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28, -OC(O)R27, -OCrCβ-alky!-C(O)OR27, -SC1-C6-BlKyI-C(O)OR27, -C2-Cβ-alkenyl- C(=O)OR27. -NR27-C(=O)-C1-Ce-alkyl-C(=O)OR27, -NR^-Ct=O)-C1-C6- alkenyl-C(=O)OR27-. -C(=O)NR2r-CrC6-alkyl-C(=O)OR27. -C1-Ca-alkyl-C(=O)OR27. or -C(O)OR27,
• C-t-Ce-alkyl, C2-Cβ-alkenyl or CrCβ-alkynyl.
which may optionally be substituted with one or more substituents independently se- lected from R29
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Ce-alkoxy, aryl-d-Cβ-alkyl, aryl-Qr Ce-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-Ci-C6-alkyl, heteroa PyI-C2-C6- alkenyl or heteroaryl-C2-C8-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
In another embodiment R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R26, -SR27. -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27. -OCrCe-alkyl-C(O)OR27, -Sd-Ce-alkyl-CtOJOR27, -CrCβ-alkenyl- Ct=O)OR27, -C(=O)NR27-C1-Cβ-aikyl-C(=O)OR27, -d-Qralkyl-C^OJOR27, or -C(O)OR27,
• Ci-Cβ-alkyl optionally substituted with one or more substituents independently se¬ lected from R29
• aryl, aryloxy, aroyl, aryl-CrCβ-alkoxy, aryl-Ci-C6-alkyl, heteroaryl, heteroaryl-Ci-C6- alkyl. of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
In another embodiment R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27. -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -OCrC6-aIkyl~C(OpR27,
Figure imgf000025_0001
-Ca-Cβ-alkenyl- C(=O)OR27, -C(=O)NR27-C1-Cβ-alkyl-C(=O)OR27, -C1-Cβ-alkyl-C(=O)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
•aryl, aryloxy, aroyl, aryl-Ci-Ce-alkoxy, aryl-Ci-Cβ-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30. In another embodiment R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28,
-OC(O)R27, -OCrCβ-alkyl-CtOPR27, -Sd-Cβ-alkyl-C(O)OR27, -C;rCβ:alkenyl- CC=O)OR27, -C(=O)NR2r-Ct-C6-alkyl-C(=O)OR27, -C1-Cβ-alkyl-C(=O)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
•AΓG1 , ArGI-O-, ArGI-C(O)-, ArG1-d-Ce-alkoxy, ArG1-d-Cβ-alkyl, Het3, Het3-Cr Cβ-alkyJ of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30. In another embodiment R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28. -SR27, -NR27C(O)R2β, -NR27C(O)OR28. -OC(O)R27, -OC1-C6-alkyl-C(O)OR27, -SCi-Ce-alkyl-C(O)0R27, -C2-C6-alkenyl- Cf=O)OR27, -C(=O)NR27-C1-Cβ-alkyI-C(=0)0R27, -CrCβ-alkyl-C(=O)OR27, or -C(O)OR27,
• CrCβ-alkyl optionally substituted with one or more substituents independently se- lected from R29
• phenyl, phenyloxy, phenyl-CrC6-alkoxy, phenyl-CrCβ-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30. In another embodiment R19 is hydrogen or methyl. In another embodiment R1Θ is hydrogen. In another embodiment R87 is Hydrogen, CrCβ-aIkyl or aryl. In another embodiment R27 is hydrogen or CrCβ-alkyl. In another embodiment R28 is hydrogen or d-Cs-alkyl. In another embodiment F is a valence bond.
In another embodiment F is CrCβ-alkylene optionally substituted with one or more hydroxy, Ci-Cβ-alkyl, or aryl.
In another embodiment G is CrCβ-alkylene or arylene, wherein the arylene is optionally sub¬ stituted with up to three substituents R24, R25 and R28. In another embodiment G is Ci-C6-alkylene or ArG1, wherein the arylene is optionally substi¬ tuted with up to three substituents R24, R28 and R2β In another embodiment G is Ci-Cβ-alkylene.
In another embodiment G is phenylene optionally substituted with up to three substituents R24, R25 and R26. In another embodiment R24, R25 and R26 are independently selected from
• hydrogen, halogen, -CHF2. -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - NO2, -OR27, -NR27R26, -SR27. -C(O)NR27R28, -OC(O)NR27R28. -NR27C(O)R28. -NR27C(O)OR28, -CH2C(O)NR27R28. -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28, -OC(O)R27, -OCrCe-alkyl-C(O)OR27, -SCrC6~alkyl-C(O)OR2r, -CrC6-alkenyl-
C(=O)OR27, -NRa7-C(=O)-CrC6-alkyl-C(=0)OR27, -NR27-C(=O)-CrCβ- alkenyl-C(=O)OR27-. -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -C,-Cβ-alkyl-C(=O)OR27. or -C(O)OR27,
"CrCe-alkyl, C2-CB-alkenyl or CrCβ-alkynyl, which may optionally be substituted with one or more substituents independently se¬ lected from R29
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-CrCe-alkoxy, aryl-Ci-Cθ-alkyl, aryl-Cr Cβ-alkenyl, aryl-C2-Cβ-alkynyl, heteroaryl, heteroaryl-Ci-C6-alkyl, heteroaryl-C2-Ce- alkenyl or heteroaryl-CrC6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
In another embodiment R24, R25 and R26 are independently selected from
•hydrogen, halogen, -OCF3, -OR27, -NR27R28. -SR27, -NR27C(O)R28. -NR27C(O)OR28. -OC(O)R27, -Od-Cβ-alkyl-CtOJOR27,
Figure imgf000027_0001
-C2-C6-a!kenyi- C(=O)OR27, -C(=O)NR27-CrCβ-alkyl-C(=O)OR27.
Figure imgf000027_0002
or
-C(O)OR27,
• Ct-Cβ-alkyt, Cj-Ce-alkenyl or C2-Cθ-alkyπyl,
which may optionally be substituted with one or more substituents independently se¬ lected from R29
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Cβ-alkoxy, aryl-Ci-Cβ-alkyl, aryl-C≥- Ce-alkenyl, aryl-Cz-Ce-alkynyl, heteroaryl, heteroaryl-d-Cβ-alkyl, heteroaryl-C2-Cβ- alkenyl or heteroaryl-CrCβ-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
In another embodiment R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -OCrCe-alkyl-CpjOR27, -SC1-C6-^KyI-C(O)OR27, -CrC6-alkenyl-
Figure imgf000027_0003
or -C(O)OR27, •Ci-Cβ-alkyl optionally substituted with one or more substituents independently se¬ lected from R29
•aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-d-Cβ-alkyl, heteroaryl, heteroaryl-d-Cβ- alkyl.
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
In another embodiment R24, R25 and R26 are independently selected from
•hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28. -OC(O)R27, -OC1-Ce-BlKyI-C(O)OR27, -SCrCβ-alkyl-C(O)OR27, -Cz-Ce-alkenyl- C(=O)0R27, -C(=O)NR27-C,-Cβ-alkyl-C(=O)OR27, -C1-Ce-alkyl-C(=O)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
-ArGI, ArGI-O-, ArGI-C(O)-, ArGI-d-Cβ-alkoxy, ArG1-d-C6-alkyl, Het3, Het3-d-
Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30. In another embodiment R24, R2S and R28 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -OCrCβ-alkyl-C(O)OR27, -Sd-Cβ-alkyl-CtOJOR27, -CrCβ-alkenyl- Ct=O)OR27, -C{=O)NR27-CrCβ-alkyl-C(=O)OR27, -C1-Cβ-alkyl-C(=O)OR27, or -C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
•ArG1, ArGI-O-, ArGI-C(O)-, ArG1-CrCβ-alkoxy, ArG1-CrC6-alkyl, Het3, Het3-d- Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30. In another embodiment R24, R25 and R28 are independently selected from
.hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR26,
-OC(O)R27, -OCrCβ-alkyl-CζOJOR27, -SC1-Ce-SIlCyI-C(O)OR27, -C2-Ce-alkenyl- C(=O)OR27, -C(=O)NR27-CrCe-alkyl-C(=O)0R27, -CrCβ-a!kyl-C(=O)OR27, or -C(O)OR27.
•methyl, ethyl propyl optionally substituted with one or more substituents independ- ently selected from R29
•ArG1, ArGI-O-, ArGI-d-Cβ-alkoxy. ArG1-Ci-C6-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30.
In another embodiment R20 is hydrogen or methyl.
In another embodiment R20 is hydrogen.
In another embodiment R27 is hydrogen, Ci-C6-alkyl or aryl.
In another embodiment R27 is hydrogen or CrC6-alkyl or ArG1. In another embodiment R27 is hydrogen or C1-Cβ-alkyl.
In another embodiment R28 is hydrogen or d-Ce-alkyl.
In another embodiment R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3. -OCF3, -NO2, -OR27, -NR27R28, -SR27, -S(O)R27, -S(O)2R27, -C(O)NR27R29, -CH2OR27, -OC(O)R27, -0CrCβ-a!kyl-C(O)0R27, -SC1-C8- alkyl-C(O)OR27, Or -C(O)OR27,
• d-Cβ-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, optionally substituted with one or more substituents independently selected from R29
• aryl, aryloxy, aroyl, aryl-Ci-Cβ-alkoxy, aryl-d-Cβ-alkyl, heteroaryl, heteroaryl-CrCβ- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
In another embodiment R17 and R18 are independently selected from •hydrogen, halogen, -CN. -CF3, -NO2, -OR27, -NR27R28, or -C(O)OR27,
•Ci-Cβ-alkyl optionally substituted with one or more substituents independently se- lected from R29
• aryl, aryloxy, aroyl, aryl-Ci-Cβ-alkoxy, aryl-CrCe-alkyl, heteroaryl, heteroaryl-CrC6- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
In another embodiment R17 and R1βare independently selected from
• hydrogen, halogen, -CN, -CF3, -NO2, -OR27, -NR27R28. or -C(O)OR27 • methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R2B
•aryl, aryloxy, aroyl, aryl-C,-Cβ-alkoxy, aryl-Ci-Ce-alkyl, heteroaryl, heteroaryl-Ci-Cβ- alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
In another embodiment R17 and R18 are independently selected from
• hydrogen, halogen, -CN1 -CF3, -NO2, -OR27, -NR27R28, or -C(O)OR27
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29 »ArG1, ArGI-O-. ArGI-C(O)-, ArGI-d-Cβ-alkoxy, ArGI-d-Cs-alkyl, Het3. Het3-C,-
Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30. In another embodiment R17 and R18 are independently selected from • hydrogen, halogen, -CN, -CF3, -NO2, -OR27. -NR27R28, or -C(O)OR27
• Ct-Cβ-alkyl optionally substituted with one or more substituents independently se¬ lected from R29
• phenyl, phenyloxy.
Figure imgf000030_0001
phenyl-C,-Cβ-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se- lected from R30.
In another embodiment R27 is hydrogen or C,-Cβ-alkyl. In another embodiment R27 is hydrogen, methyl or ethyl. In another embodiment R28 is hydrogen or Ci-C6-alkyl. In another embodiment R28 is hydrogen, methyl or ethyl. In another embodiment R72 is -OH or phenyl. In another embodiment CGr is
Figure imgf000031_0001
In another embodiment CGr is of the form H-I-J-
Figure imgf000031_0002
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R31
I is selected from *a valence bond,
• -CH2N(R32)- or -SO2N(R33)-,
Figure imgf000031_0003
Z1 is S(O)2 or CH2. Z2 is -NH-, -O-or -S-, and n is 1 or 2,
J is
• Ci-Cfi-alkylene, CrCe-alkeπylene or CrCe-aikynylene, which may each optionally be substituted with one or more substituents selected from R34,
• Arylene, -aryloxy-. arylene-oxycarbonyl-, -aroyl, arylene-CrCβ-alkoxy-, ary- lene-CrCβ-alkylene, arylene-CrC6-alkenylene, arylene-C2-Ca-alkynylene, heteroary- lenβ,
Figure imgf000031_0004
heteroarylene-Ca-Ce-alkenylene or heteroary- leπe-C2-C6-alkynylene, wherein the cyclic moieties are optionally substituted with one or more substituents selected from R37, R31 is independently selected from hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3. -OCF2CHF2, -S(O)2CF3, -SCF3, -NO2, -OR33, -C(O)R35, -NR35R36. -SR35, -NR35S(O)2R38, -S(O)2NR35R36, -S(O)NR35R36. -S(O)R35, -S(O)2R35, -C(O)NR35R36, -OC(O)NR35R36, -NR35C(O)R36, -CH2C(O)NR35R38, -OCH2C(O)NR35R38, -CH2OR33, -CH2NR35R36. -OC(O)R35, -Od-Ce-alkyl-C(O)OR35, -SCi-Ce-alkyl-CtOJOR35 -C-rOralkenyl-
Figure imgf000032_0001
-NR36-C(=O)-C1-Cβ-alkenyl-C(=O)OR35-, d-Cβ-alkyl, Ci-Cβ-alkanoyl or -C(O)OR35,
R32 and R33 are independently selected from hydrogen, CrCβ-alkyl or Ci-Ce-alkanoyl,
R34 is independently selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R38,
R35 and R36 are independently selected from hydrogen, CrCe-alkyl, aryl-d-Cβ-alkyl or aryl, or R35 and R36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(OpR35, -C(O)H, -CN, -CF3, -OCF3, -NO2, - OR35. -NR35R39, d-Ce-alkyl or d-Cβ-alkanoyl.
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment H is
Figure imgf000032_0002
In another embodiment H is
Figure imgf000032_0003
In another embodiment H is
Figure imgf000033_0001
In another embodiment t is a valence bond, -CH2N(R32)-, or -SO2N(R33)-. In another embodiment 1 is a valence bond. In another embodiment J is
•CVCe-alkylene, C2-Cβ-alkenylene or C2-C6-alkynylene, which may optionally be substituted with one or more substituents selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R39,
• arylene, or heteroarylene, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
In another embodiment J is
• arylene or heteroarylene, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
In another embodiment J is «ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37. tn another embodiment J is
• phenylene or naphthyleπe optionally substituted with one or more substitu¬ ents independently selected from R37. In another embodiment R32 and R33 are independently selected from hydrogen or Ci-Cβ-alkyl.
In another embodiment R3* is hydrogen, halogen, -CN, -CF3, -OCF3, -SCF3, -NO2, -OR35,
-C(O)R35, -NR35R36, -SR35. -C(O)NR35R38, -OC(O)NR35R36, -NR35C(O)R38, -OC(O)R35, -OC1-
Ce-BlKyI-C(O)OR35, -Sd-C6-alkyl-C(O)OR35 or -C(O)OR35.
In another embodiment R34 is hydrogen, halogen, -CF3. -NO2, -OR35, -NR35R36, -SR35, -NR35C(O)R38, or -C(O)OR35.
In another embodiment R34 is hydrogen, halogen, -CF3, -NO2, -OR35, -NR35R36, or
-NR35C(O)R39.
In another embodiment R34 is hydrogen, halogen, or -OR35.
In another embodiment R33 and R36 are independently selected from hydrogen, CrCβ-alkyl, or aryl.
In another embodiment R35 and R36 are independently selected from hydrogen or C|-Ce-alkyl.
In another embodiment R37 is halogen. -C(O)OR35, -CN. -CF3, -OR35, -NR35R36, Ci-Ce-alkyl or
Ci-C6-alkanoyl. In another embodiment R37 is halogen, -C(O)OR35, -OR35, -NR35R38, d-Cβ-alkyl or C1-C6- alkanoyl.
In another embodiment R37 is halogen, -C(O)OR35 or -OR35.
In another embodiment CGr is
Figure imgf000034_0001
wherein K is a valence bond, CrC6-alkylene, -NH-C(=O)-U-, -Ci-Cβ-alkyi-S-, -CrCβ-alkyl-O-, -C(=O)-f or -Cf=O)-NH-. wherein any Ci-Cβ-alkyl moiety is optionally substituted with R =>338
U is a valence bond, d-Ce-alkenylene, -Ct-C6-alkyl-O- or Ci-Cβ-alkyJeπe wherein any C,- Ce-alkyl moiety is optionally substituted with d-Cβ-alkyl,
R38 is CrCβ-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R39,
R3B is independently selected from halogen, cyano, nitro, amino,
M is a valence bond, arylene or heteroaryfene, wherein (he aryl or heleroaryl moieties are optionally substituted with one or more substituents independently selected from R*0,
R40 is selected from
• hydrogen, halogen. -CN, -CH2CN, -CHF2. -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -OS(O)2CF3. -SCF3, -NO2. -OR41. -NR41R42. -SR41. -NR41S(O)2R42, -S(O)2NR41R42, -S(O)NR41R42. -S(O)R41, -S(O)2R41, -OS(O)2 R41, -C(O)NR41R42, -OC(O)NR41R42, -NR41C(O)R42, -CH2C(O)NR41R42, -OCrCβ- alkyl-C(O)NR41R42. -CH2OR41, -CH2OC(O)R41, -CH2NR41R42, -OC(O)R41, -OC1-C8- alkyl-C(O)OR4\ -OCrCe-alkyl-OR41, -S-CrC6-alkyl-C(O)OR4\ -Cz-Cβ-alkenyl- C(O)OR41, -NR41-C(=O)-CrC6-alkyl-C(=O)OR41,
Figure imgf000034_0002
alkenyl-C(=0)OR41 , -C(O)OR41, -C2-C6-alkeπyl-C(=O)R41, =O, -NH-C(=O)-O-C,- Ce-alkyl, or -NH-C(=O)-C(=O)-O-C,-C8-alkyl,
• CrC6-alkyl, C2-C6-alkenyl or CrCβ-alkynyl, which may each optionally be substituted with one or more substituents selected from R43, •aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryI-d-C6-alkoxy, aryl-Ci-C6-alkylr ary[-Cz-Cβ-aIkenyl, aroyl-C2-Cβ-alkenyl, aryl-Cz-Cβ-alkynyl, heteroaryl, heteroaryl-Ci- Cβ-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-Cβ-alkyπylt wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44,
R41 and R42 are independently selected from hydrogen, -OH1 d-Cβ-alkylf d-CB-alkenyl, aryl- d-Cβ-alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R45, and the aryl moieties may optionally be substi- tuted with one or more substituents independently selected from R46; R41 and R42 when at¬ tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R43 is independently selected from halogen, -CN, -CF3, -OCF3, -OR41, and -NR41R42
R44 is independently selected from halogen, -C(O)OR41, -CH2C(O)OR41, -CH2OR41, -CN, - CF3. -OCF3, -NO2. -OR41, -NR41R42 and CrCθ-alkyl, R45 is independently selected from halogen. -CN, -CF3, -OCF3. -O-C-Ce-alkyl, -C(O)-O-C1- Cβ-alkyl, -COOH and -NH2,
R4β is independently selected from halogen, -C(O)Od-Cβ-alkyl, -COOH, -CN. -CF3, -OCF3, - NO2, -OH, -OCrCβ-alkyl, -NH2, C(=0) or d-Cβ-alkyl,
Q is a valence bond, d-Cβ-alkylene, -d-Ce-alkyl-O-, -d-Cθ-alkyl-NH-. -NH-Ci-Cβ-alkyl, -NH-C(O)-, -C(=0)-NH-. -O-d-Cβ-alkyl, -C(Oh or -d-Cβ-alkyl-C(=O)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48,
R47 and R48 are independently selected from hydrogen, CrCe-alkyl, aryl optionally substituted with one or more R49,
R49 is independently selected from halogen and -COOH,
T is •CrCβ-alkylene, CrCβ-alkenylene , C2-Cβ-alkynylene, -CrCβ-alkyloxy-carbonyl. wherein the alkylene, alkenylene and alkynylene moieties are optionally substituted with one or more substituents independently selected from R50, •arylene, -aryloxy-, -aryloxy-carbonyl-, arylene-d-Cβ-alkylene, -aroyl-, arylene-Ci- Cβ-alkoxy-, arylene-CrCβ-alkenylene, arylene-C2-Ce-alkynylene, heteroarylene, het- eroarylene-Ci-Cβ-alkylene, heteroarylene-C2-Ce-alkenylene, heteroarylene-C2- Cβ-alkynylene,
wherein any alkylene, alkenylene , alkynylene. arylene and heteroarylene moiety is optionally substituted with one or more substituents independently selected from R50,
R50 is CrCβ-alkyl, C,-Cβ-alkoxy. aryl, aryloxy. aryl-d-Cβ-alkoxy. -C(=O)-NH-Ci-Cβ-alkyl-aryl, heterαaryl, heterαaryl-d-Ce-alkoxy, -Ci-Cβ-alkyl-COOH, -O-Ci-Cβ-alkyl-COOH, -S(O)2R51, -QrCβ-alkenyl-COOH, -OR51, -NO2, halogen, -COOH1 -CF3, -CN, =0, -N(R51R52), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53,
R51 and R52 are independently selected from hydrogen and d-Ce-alkyl,
R53 is independently selected from Ci-C6-alkyl. d-Ce-alkoxy, -Ci-C6-alkyl-COOH, -Cr
Cβ-alkenyl-COOH, -OR51, -NO2, halogen, -COOH, -CF3, -CN. Or-N(R51R52),
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment K is a valence bond. Ci-Cβ-alkylene, -NH-C(=O)-U-. -Ci-Cβ-alkyl-S-,
-CrCβ-alkyl-O-, or -C(=O)-. wherein any Ci-C6-alkyl moiety is optionally substituted with R38. In another embodiment K is a valence bond, CrCβ-alkylene, -NH-C(=O)-U-, -CrCβ-alkyl-S-, or -Ci-Cfj-alkyl-O, wherein any CrCβ-alkyl moiety is optionally substituted with R38.
In another embodiment K is a valence bond. Ci-CB-alkylene, or -NH-C(=O)-U, wherein any
CrCβ-alkyl moiety is optionally substituted with R38.
In another embodiment K is a valence bond or d-Cβ-alkylene, wherein any d-Cβ-alkyl moi- ety is optionally substituted with R38.
In another embodiment K is a valence bond or -NH-C(=O)-U.
In another embodiment K is a valence bond.
In another embodiment U is a valence bond or -C,-Ce-alkyl-O-.
In another embodiment U is a valence bond In another embodiment M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
In another embodiment M is ArG1 or Het1, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
In another embodiment M is ArGI or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40. In another embodiment M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40. In another embodiment M is phenylene optionally substituted with one or more substituents independently selected from R40.
In another embodiment M is indolylene optionally substituted with one or more substituents independently selected from R40. In another embodiment M is
Figure imgf000037_0001
In another embodiment M is carbazolylene optionally substituted with one or more substitu¬ ents independently selected from R40. In another embodiment M is
Figure imgf000037_0002
In another embodiment R40 is selected from
• hydrogen, halogen, -CN1 -CF3, -OCF3, -NO2, -OR41, -NR41R42, -SR41, -S(O)2R41, -NR41C(O)R42, -0C1-Cθ-alkyl-C(O)NR4V2, -QrCβ-alkenyl-C(=O)OR41. -C(O)OR41, =O,
Figure imgf000037_0003
or-NH-C(=O)-C(=O)-O-C1-Cβ-alkyl,
Ci-Cβ-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• aryl, aryloxy, aryl-Ci-Cβ-alkoxy, aryl-C1-C6-alkyl, aryl-Cz-Ce-alkenyl, heteroaryl, het- eroaryl-d-Ce-alkyl, or heteroaryl-C2-Ce-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
In another embodiment R40 is selected from •hydrogen, halogen, -CN, -CF3, -OCF3, -NO2, -OR41, -NR41R42, -SR41, -S(O)2R41, -NR41C(O)R42, -OC,-Cβ-alkyt-C(O)NR41R42, -C2-Cβ-alkenyl-C(=O)OR41, -C(O)OR41.
Figure imgf000038_0001
d-Cβ-alkyl or C2-C3- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• ArG1, ArGI-O-, ArGI-d-Cβ-alkoxy, ArG1-CrC6-alkyl, ArG1-C2-Ce-alkenyl, Het3, Het3-Ci-CB-alkyl. or Het3-C2-Cβ-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
In another embodiment R40 is selected from
• hydrogen, halogen, -CF3, -NO2. -OR41, -NR41R42. -C(O)OR41, =O, or -NR41C(O)R42.
• C,-Cβ-alkyl,
• ArG1. In another embodiment R40 is selected from
• Halogen, -NO2, -OR41, -NR41 R42, -C(O)OR41. or -NR41C(O)R42.
• Methyl,
• Phenyl.
In another embodiment R41 and R42 are independently selected from hydrogen, d-Ce-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
In another embodiment R41 and R42 are independently selected from hydrogen, methyl, ethyl. or phenyl, wherein the phenyl moieties may optionally be substituted with hafogen or -
COOH.
In another embodiment Q is a valence bond, d-Cβ-alkylene, -d-C6-alkyl-O-, -Ci- C8-alkyl-NH-, -NH-d-Cβ-alkyl, -NH-C(=O)-. -C(=O)-NH-. -O-CrC6-alkyl, -C(=0)-, or -C,-
Cβ-alkyl-C(=O)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R*8.
In another embodiment Q is a valence bond, -CH2-. -CH2-CH2-, -CH2-O-, -CH2-CH2-O-,
-CH2-NH-, -CH2-CH2-NH-, -NH-CH2-, -NH-CH2-CH2-, -NH-C(O)-, -C(O)-NH-, -0-CH2-, -0-CH2-CH2-. or -C(O)-.
In another embodiment R4r and R48 are independently selected from hydrogen, methyl and phenyl.
In another embodiment T is
• d-Cβ-alkylene optionally substituted with one or more substituents independently selected from R50, • arylene, arylene-CrC^-alkylene, heteroarylene, wherein the alkylene, arylene and heteroaryleπe moieties are optionally substituted with one or more substituents inde¬ pendently selected from R50.
In another embodiment T is #Ci-Cβ-alkylene optionally substituted with one or more substituents independently selected from R50,
• ArGI1 ArGI-d-Cβ-alkylene, Het3, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from Rw.
In another embodiment T is
• Ci-Ce-alkylene, optionally substituted with one or more substituents independently selected from R50,
•phenylene, phenylene-d-Ce-alkylene, wherein the alkylene and pheπylene moieties are optionally substituted with one or more substituents independently selected from R50.
In another embodiment R50 is d-Cβ-alkyl, Ci-Cβ-alkoxy. aryl, aryloxy. aryl-d-Ce-alkoxy, -C(=O>NH-C,-C6-alkyl-aryl, heteroaryl, -CrCe-alkyl-COOH, -O-CrCβ-alkyl-COOH. -S(O)2R51, -CrCβ-alkenyl-COOH, -OR51, -NO2, halogen. -COOH, -CF3, -CN, =O, -N(R51R52), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53.
In another embodiment R50 is d-Cβ-alkyl, d-Cβ-alkoxy, aryl, aryloxy, aryl-d-Cβ-alkoxy , -OR61, -NO2, halogen, -COOH1 -CF3, wherein any aryl moiety is optionally substituted with one or more R53. In another embodiment R50 is d-Ce-alkyl, aryloxy, aryl-Ci-C6-alkoxy , -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53.
In another embodiment R50 is CrC8-alkyl, ArGI-O-, ArG1-CrCe-alkoxy , -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53. In another embodiment R50 is phenyl, methyl or ethyl. In another embodiment R50 is methyl or ethyl. In another embodiment R51 is methyl.
In another embodiment R53 is d-Ce-alkyl, d-C6-alkoxy, -OR51, halogen.or -CF3.
In another embodiment CGr is
Figure imgf000040_0001
wherein V is Ci-Cβ-a1kylene, arylene, heteroarylene, arylene-Ci-e-alkylene or aryfene-C2^- alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R55,
R54 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R55 is independently selected from -hydrogen, halogen. -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3,
-OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3, -NO2, -OR56. -NR58R57, -SR56. -NR56S(O)2R57, -S(O)2NR56R67, -S(O)NR56R57, -S(O)R59, -S(O)2R56. -OS(O)2 R» -C(O)NR56R57, -OC(O)NR50R57, -NR56C(O)R57, -CH2C(O)NR56R57. -OC1-C6- 8^yI-C(O)NR56R57, -CH2OR56, -CH2OC(O)R58, -CH2NR56R57. -OC(O)R58, -OC1-C8- alkyl-CtOJOR59,
Figure imgf000040_0002
-SCrCβ-alkyl-CCOJOR56, -Cz-Cβ-alkenyl-
C(=0)0Rse, -NR56-C(=O)-C,-C6-alkyl-C(=O)OR5a. -NRsβ-C(=O)-CrCe- alkenyl-C(=O)OR56 , -C(O)OR56, or-C2-Cβ-alkenyl-C(=O)R56,
• CrCβ-alkyl, C2-Cβ-alkenyl or C2-C6-alkynyl,
which may optionally be substituted with one or more substituents selected from R58,
•aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl. aryl-CrCβ-alkoxy, aryl-CrCe-alkyl, aryl-C2-Cβ-alkenyl, aroyl-C2-Ce-alkenyl. aryl-C2-Ce-alkynyl, heteroaryl, heteroaryl-Cr Cβ-alkyl, heteroaryl-CrCe-alkenyl or heteroaryl~Cz-Cβ-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R59,
R56 and R57 are independently selected from hydrogen, OH1 CF3, Ci-C12-alkyl, ary[-C,-Cβ- alkyl, -C(=O)-Ci-Cβ-alkyl or aryl, wherein the alky! groups may optionally be substituted with one or more substituents independently selected from R60, and the aryl groups may option¬ ally be substituted with one or more substituents independently selected from R61; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R58 is independently selected from hafogeπ, -CN, -CF3, -OCF3, -OR56, and -NR56R57,
R59 is independently selected from halogen. -C(O)OR56. -CH2C(O)OR56, -CH2OR59, -CN, - CF3, -OCF3, -NO2, -OR56, -NR56R57 and d-Qralkyl,
R60 is independently selected from halogen, -CN, -CF3, -OCF3, -OCrCβ-alkyl, -C(O)OC1-C6- alkyl. -C(=O)-R62. -COOH and -NH2.
Rβ1 is independently selected from halogen, -C(O)OCi -Cβ-alkyl, -COOH, -CN, -CF3, -OCF3, - NO2, -OH1 -OC,-Cβ-alkyl, -NH2. C(=0) or d-Cβ-alkyl,
R62 is d-Ce-alkyl, aryl optionally substituted with one or more substituents independently se¬ lected from halogen, or heteroaryl optionally substituted with one or more CrCβ-alkyl inde¬ pendently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment V is arylene, heteroarylene, or arylene-Ci.Cβ-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected R54, and the arylene or heteroarylene is optionally substituted with one or more substituents inde- pendently selected from R55.
In another embodiment V is arylene, Het1, or aryleπe-d.Cβ-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde¬ pendently selected from R55. In another embodiment V is arylene, Het2, or arylene-d-Cβ-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents inde¬ pendently selected from R55. In another embodiment V is arylene, Het3, or arylene-d.Cε-alkylene, wherein the alkylene is optionally substituted with one or more substituents independently selected from R54, and the arylene or heteroarylene moiety is optionally substituted with one or more substituents Inde¬ pendently selected from R55.
In another embodiment V is arylene optionally substituted with one or more substituents in¬ dependently selected from R55. In another embodiment V is ArG1 optionally substituted with one or more substituents inde¬ pendently selected from R53.
In another embodiment V is phenylene, naphthyleπe or anthranylene optionally substituted with one or more substituents independently selected from R55.
In another embodiment V is phenylene optionally substituted with one or more substituents independently selected from R55.
In another embodiment R55 is independently selected from
• halogen. CrCβ-alkyl, -CN, -OCF3 ,-CF3, -NO2, -OR58, -NR56R57, -NR56C(O)R57 -SR56, -OCrCβ-alkyl-CtOJOR56, or -C(O)OR58,
• Ci-Cβ-alkyl optionally substituted with one or more substituents independently se- lected from R58
•aryl, aryl-Ci-Ce-alkyl, heteroaryl, or heteroaryl-Ci-Cθ-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents independently selected from R59. In another embodiment R55 is independently selected from • halogen, d-Cβ-alkyl, -CN. -OCF3 ,-CF3. -NO2, -OR56, -NR58R57. -NR58C(O)R57
-SR56. -OCrCe-alkyl-CpjOR56. or -C(O)OR56
• Ci-Ce-alkyl optionally substituted with one or more substituents independently se¬ lected from R58
• ArGi, ArGI-CrCβ-alkyl, Het3, or Het3-C,-Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents independently selected from R59.
In another embodiment R55 is independently selected from halogen, -OR59, -NR58R57, -C(O)OR58, -OCrC-alkyl-CfOJOR56, -NR58C(O)R57 or CrCθ-alkyl. In another embodiment R55 is independently selected from halogen, -OR56, -NR58R57. -C(O)OR58, -OC-Cβ-alkyl-qojOR56, -NR58C(O)R57, methyl or ethyl. In another embodiment R56 and R57 are independently selected from hydrogen, CF3, d-Cia-alkyl, or -C(=O)-C,-Cβ-alkyl; R58 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom. tn another embodiment R56 and RS7 are independently selected from hydrogen or d-C12-alkyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 mem- bered heterocyclic ring with the said nitrogen atom.
In another embodiment R56 and R57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
In another embodiment CGr is
Figure imgf000043_0001
wherein AA is Ci-Cβ-alkylene, arylene, heteroarylene, arylene-Ci-C6-alkylene or arylene-Cj. Cβ-alkenylene, wherein the alkylene or alkenylene is optionally substituted with one or more substituents independently selected from R63, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R84,
R63 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2,
R6* is independently selected from
•hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2. -OCH2CF3,
-OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3. -NO2, -OR65, -NR65R66, -SRβ5, -NR65S(O)2R60, -S(O)2NR65R86, -S(O)NR65R68, -S(O)R65. -S(O)2R65, -OS(O)2 R65, -C(O)NR65R96. -OC(O)NR65R66, -NR65C(O)R86, -CH2C(O)NR65R66. -OCi-C6- 3IKyI-C(O)NR65R66, -CH2OR65, -CH2OC(O)R65, -CH2NR65R66, -OC(O)R65, -OC1-C8- alkyl-CfOJOR65, -OCrC6-alkyl-OR65, -Sd-Cβ-alkyl-CfOJOR65, -Ca-Ce-alkenyl-
C(=O)OR85, -NRfl5-C(=O)-C1-Cβ-alkyl-C(=O)OR65, -NR65-C(=O)-C,-Cβ- alkenyl-C(=O)OR65 , -C(O)OR65, or -C2-C6-alkenyl-C(=O)R65,
* Ci-Cβ-alkyl, C2-Cβ-alkenyl or C2-C6-alkynyl. each of which may optionally be substi- tuted with one or more substituents selected from Rθ7, • aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-CrC6-alkoxy, aryl-Ci-CB-alkyl, aryl-Cz-Ce-alkeπyl, aroyl-Cz-Cβ-alkenyl, aryl-C2-CB-alkynyl, heteroaryl, heteroaryl-d- Cβ-alkyl, heteroaryl-CjrCe-alkenyl or heteroaryl-C2-C8-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R6*,
Rβ5 and Rβθ are independently selected from hydrogen, OH. CF3, Ci-C12-alkyl, aryl-Ci-Cβ- alkyl, -CC=O)-R69, aryl or heteroaryl, wherein the alkyl groups may optionally be substituted with one or more substituents selected from R70, and the aryl and heteroaryl groups may op¬ tionally be substituted with one or more substituents independently selected from R71; Re5 and RBβ when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R67 is independently selected from halogen. -CN, -CF3. -OCF3, -OR65, and -NR65R66.
R68 is independently selected from halogen, -C(O)OR65. -CH2C(O)OR65, -CH2OR65, -CN, - CF3, -OCF3, -NO2, -OR65, -NR65R68 and d-Ce-alkyl.
R69 is independently selected from Ci-Cβ-alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more Ci-Cθ-alkyl,
R70 is independently selected from halogen, -CN, -CF3, -OCF3, -OCrCβ-afkyl, -C(O)OC1-Ce- alkyl, -COOH and -NH2,
R71 is independently selected from halogen, -C(O)OCi-Cβ-alkyl, -COOH, -CN, -CF3, -OCF3, -
NO2. -OH, -OCi-Cβ-alkyl. -NH2, C(=0) or Ci-Cβ-alkyl,
or any enantiomer, diaεtereαmer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
In another embodiment AA is arylene, heteroarylene or arylene-d.Cβ-alkylene, wherein the alkylene is optionally substituted with one or more R63, and the arylene or heteroarylene is optionally substituted with one or more substituents independently selected from R64. In another embodiment AA is aryleπe or heteroarylene, wherein the aryiene or heteroarylenβ is optionally substituted with one or more substituents independently selected from R64. In another embodiment AA is ArG1 or Het1 optionally substituted with one or more substitu¬ ents independently selected from R64. In another embodiment AA is ArG1 or Het2 optionally substituted with one or more substitu¬ ents independently selected from R84.
In another embodiment AA is ArG 1 or Het3 optionally substituted with one or more substitu¬ ents independently selected from R64. In another embodiment AA is phenylene, naphtylene, anthrylene, carbazolylene, thienylene, pyridylene, or benzodioxylene optionally substituted with one or more substituents independ¬ ently selected from R64.
In another embodiment AA is phenylene or naphtylene optionally substituted with one or more substituents independently selected from R64. In another embodiment R64 is independently selected from hydrogen, halogen, -CF3, -OCF3, -OR65, -NR65R66, C-Ce-alkyl, -OC(O)R65, -OC1-C6-BlRyI-C(O)OR65,
Figure imgf000045_0001
aryloxy or aryl, wherein Ci-CB-alkyl is optionally substituted with one or more substituents independ¬ ently selected from R97, and the cyclic moieties optionally are substituted with one or more substituents independently selected from R88. In another embodiment R84 is independently selected from halogen, -CF3, -OCF3, -OR65, -NR65R66, methyl, ethyl, propyl, -OC(O)R65, -OCH2-C(O)OR65, -OCH2-CH2-C(O)OR65, phenoxy optionally substituted with one or more substituents independently selected from R68.
In another embodiment R65 and ReB are independently selected from hydrogen, CF3, C^C^-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents inde- pendently selected from R71.
In another embodiment R65 and R8β are independently hydrogen, Ci-C1z-alkyl, aryl, or het¬ eroaryl optionally substituted with one or more substituents independently selected from R7\ In another embodiment R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents inde- pendently selected from R71.
In another embodiment R65 and Rββ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het2 optionally substituted with one or more substituents inde¬ pendently selected from R71. In another embodiment R65 and Rββ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG 1 or Het3 optionally substituted with one or more substituents inde¬ pendently selected from R71. In another embodiment R65 and Reβ are independently hydrogen, methyl, ethyl, propyl, butyl,
2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R71 independently; or isoxazolyl optionally substituted with one or more substituents independ¬ ently selected from R71. In another embodiment R71 is halogen or Ci-Cβ-alkyl.
In another embodiment R71 is halogen or methyl.
In another embodiment Frg consists of 0 to 5 neutral amino acids independently selected from the group consisting of GIy, Ala, Thr, and Ser.
In another embodiment Frg consists of 0 to 5 GIy. In another embodiment Frg consists of O GIy.
In another embodiment Frg consists of 1 GIy.
(n another embodiment Frg consists of 2 GIy.
In another embodiment Frg consists of 3 GIy.
In another embodiment Frg consists of 4 GIy. In another embodiment Frg consists of 5 GIy.
In another embodiment GB is of the formula B'-B^CKO}-, B'-B^SOz- or B1^-CH2-, wherein
B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1-B2-C(O)-. B1^-SO2- or B'-B^NH-, wherein
B1 and B2 are as defined in claim 1. In another embodiment GB is of the formula B1-B2-C(O)-, B^B2OH2- or BT-B2-NH-t wherein
B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1-B2-CH2-, B1-B2-Sθ2- or B1-B2-NH-. wherein B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1-B2-C(O)- or B1^-SO2-, wherein B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1^-C(O)- or B1-B2-CH2-, wherein B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1-B2-C(O)- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1. In another embodiment GB is of the formula Bn-B2-CH2- or B1-B2-SO2- , wherein B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1-B2-NH- or B1-B2-SO2- , wherein B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B'-B^CHz- or B1-B2-NH- , wherein B1 and B2 are as defined in claim 1.
In another embodiment GB is of the formula B1-B2-C(O)-. In another embodiment GB is of the formula B1-B2-CH2-. In another embodiment GB is of the formula B1-B2-SO2-. In another embodiment GB is of the formula B1-B2-NH-. In another embodiment B1 is a valence bond, -O-, or -S-. In another embodiment B1 is a valence bond, -O-. or -N{R6)-. In another embodiment B1 is a valence bond, -S-, or -N(R6)-. In another embodiment B1 is -O-, -S- or -N(R6)-. In another embodiment B1 is a valence bond or -O-. In another embodiment B1 is a valence bond or -S-. In another embodiment B1 is a valence bond or -N(R6)-. In another embodiment B1 is -O-or -S-. In another embodiment B1 is -O-or -N(R6)-. In another embodiment B1 is -S-or -N(R6)-. tn another embodiment B1 is a valence bond. In another embodiment B1 is -O-. In another embodiment B1 is -S-. In another embodiment B1 is -N(R8)-.
In another embodiment B2 is a valence bond, d-Ctβ-alkylene, C2-Cia-alkenylene, C2-Ci8- alkynylene. arylene, heteroarylene. -CrC,a-alkyl-ary1-, -C(=O)-Ci-Cia-alkyl-C(=O)-t -C(=O>- C1-C1β-alkyl-O-C1-C18-alkyl-C(=O)-, -C(=0)-C,-C,8-alkyl-S-C1-C1β-alkyl-C(=O)-, -Cf=O)-C1- C1β-alkyl-NRθ-CrC1B-alkyl-C(=O)-; and the alkylene and arylene moieties are optionally sub¬ stituted as defined in claim 1.
In another embodiment B2 is a valence bond, CrCi8-alkylene, Cz-dβ-alkenylene, C2-Ci8- alkynylene, arylene. heteroarylene, -d-C^-alkyl-aryl-. -C(=O)-C,-de-alkyl-C(=O)-, -C(=O)- CrC^-alkyl-O-Crdβ-alkyl-C^O)-, and the alkylene and arylene moieties are optionally sub¬ stituted as defined in claim 1.
In another embodiment B2 is a valence bond, d-Ciβ-alkylene, C2-C18-alkenylene, C2-C18- alkynylene, arylene, heteroarylene, -Crds-alkyl-aryl-, -C(=O)-C1-C18-alkyl-C(=O)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. In another embodiment B2 is a valence bond, d-C1β-alkylene, arylene, heteroarylene, -Ci- C18-alkyl-aryl-, -C(=O)-d-dβ-alkyl-C(=O)-, and the alkylene and arylene moieties are option¬ ally substituted as defined in claim 1.
In another embodiment B2 is a valence bond, Ci-d8-alkylene, arylene, heteroarylene, -Ci- Ci8-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. In another embodiment B2 is a valence bond, d-Ciβ-alkylene, arylene, -d-C^alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1. In another embodiment B2 is a valence bond or
Figure imgf000048_0001
and the alkylene moieties are optionally substituted as defined in claim 1. In another embodiment the insulin is selected from the goup consisting of human in¬ sulin, an analogue thereof, a derivative thereof and combinations of any of these.
In another embodiment the insulin is human insulin.
In another embodiment the insulin is an analogue of human insulin.
In another embodiment the insulin is a derivative of human insulin. In another embodiment the insulin is an analogue of human insulin wherein position
B28 is Asp, GIu, Lys, Leu, VaI, or Ala.
In another embodiment the insulin is an analogue of human insulin wherein position B28 is Asp, GIu or Lys
In another embodiment the insulin is an analogue of human insulin wherein position B28 is Asp or GIu.
In another embodiment the insulin is an analogue of human insulin wherein position B28 is Asp.
In another embodiment the insulin is an analogue of human insulin wherein position B28 is GIu. In another embodiment the insulin is an analogue of human insulin wherein position
B29 is Pro, Asp or GlU/
In another embodiment the insulin is an analogue of human insulin wherein position B29 is Pro or GIu.
In another embodiment the insulin is an analogue of human insulin wherein position B29 is Pro.
In another embodiment the insulin is an analogue of human insulin wherein position B29 is GIu.
In another embodiment the insulin is an analogue of human insulin wherein position B28 is Asp or Lys, and position B29 is Lys or Pro. In another embodiment the insulin is an analogue of human insulin wherein position
B9 is Asp or GIu.
In another embodiment the insulin is an analogue of human insulin wherein position B10 is Asp or GIu.
In another embodiment the insulin is an analogue of human insulin wherein position B10 is GIu. In another embodiment the insulin is an analogue of human insulin wherein position B1 is GIy.
In another embodiment the insulin is an analogue of human insulin wherein position 83 is Lys, Thr, Ser, Ala or GIn. In another embodiment the insulin is an analogue of human insulin wherein position
B3 is Lys, Thr, Ser or Ala.
In another embodiment the insulin is an analogue of human insulin wherein position B3 is Lys or Ala.
In another embodiment the insulin is an analogue of human insulin wherein position B3 is Lys.
In another embodiment the insulin is an analogue of human insulin wherein position B3 is Lys and position B29 is GIu.
In another embodiment the insulin is an analogue of human insulin wherein position B25 is deleted. In another embodiment the insulin is an analogue of human insulin wherein position
B27 is deleted.
In another embodiment the insulin is an analogue of human insulin wherein position B30 is deleted.
In another embodiment the insulin is an analogue of human insulin wherein position A18 is Gln.
In another embodiment the insulin is an analogue of human insulin wherein position A21 is Ala, GIn, GIu, GIy, His, lie, Leu, Met, Phe, Ser, Thr, Trp, Tyr, VaI or hSer.
In another embodiment the insulin is an analogue of human insulin wherein position A21 is Ala, GIy, lie, Leu, Phe, Ser, Thr, VaI or hSer. In another embodiment the insulin is an analogue of human insulin wherein position
A21 is Ala or GIy.
In another embodiment the insulin is an analogue of human insulin wherein position A21 is GIy.
In another embodiment the insulin is a derivative of human insulin or an analogue thereof having one or more lipophilic substituents.
In another embodiment the insulin is a derivative of human insulin or an analogue thereof wherein the Nf-amino group in position B29Lys is modified by covalent acylation with a hydrophobic moiety such as an fatty acid derivative or an litocholic acid derivative.
In another embodiment the insulin derivative is selected from the group consisting of B29-Nε-myristoyI~des(B30) human insulin, B29-NE-palmitoyl-des(B30) human insulin, B29-Nε- myristoyl human insulin, B29-Nε-palmitoyl human insulin, B28-Nε-myristoyl LysB2S Pro829 hu- man insulin. B28~Nc-palmitoyl Lysβ28 Pro829 human insulin, BSO-N^myristαyl-Thr^Lys930 hu¬ man insulin, BSO-N^palmitoyl-Thr^Lys1330 human insulin, B29-Ne-(N-palmitoyl-y-glutamyl)- des(B30) human insulin, B29-Nc-(N-lithocholyl-γ-glutamyl)-des(B30) human insulin, B29-Nε-
(ω-carboxyheptadecanoyl)-des(B30) human insulin and B29-NG-{cϋ-carboxyheptadecanoyl) human insulin. in another embodiment, the analogs of human insulin contain any combination of additional stabilizing substitutions.
In another embodiment, the analogs of human insulin contain any combination of the additional stabilizing substitutions in positions B1, B3, A18 and A21. In another embodiment the insulin is an analogue of human insulin selected from the group consisting of:
B28D desB27
B28K.B29P B3K.B29E
B29E desB25
B9E/D
B10E/D. In another embodiment the insulin is an analogue of human insulin selected from the group consisting of:
A21G
A21G, B28K, B29P
A21G. B28D A21G, B28E
A21G, B3K, B29E
A21G, desB27
A21G, B9E
A21G, B9D A21G, B10E
A21G, desB25
A21G, desB30
A21G, B28K, B29P
A21G, B28K, B29P. desB30 A21G, B28D, desB30
A21G, B28E A21G, B28E, desB30
A21G, B3K. B29E
A21G, B3K, B29E, desB30
A21G, desB27, desB30 A21G, B9E/D
A21G, B9E, desB30
A21G, B9D, desB30
A21G, B10E/D
A21G, B10E,desB30 A21G, desB25. desB30.
In another embodiment the insulin is an analogue of human insulin selected from the group consisting of:
B1G.A21G
B1G.A21G, B28K. B29P B1G.A21G, B28D
B1G.A21G.B28E
B1G.A21G. B3K, B29E
B1G,A21G.desB27
B1G, A21G, B9E B1G,A21G,B9D
B1 G, A21 G.B10E
B1G.A21GrdesB25
B1G, A21G. desB30
B1G.A21G, B28K, B29P B1G, A21G, B28K, B29P, desB30
B1G,A21G,B28D,desB30
B1G.A21G, B28E
B1G. A21G, B28E, desB30
B1G,A21G,B3K. B29E B1 G, A21 G, B3K, B29E, desB30
B1G, A21G. desB27, desB30
B1G.A21G.B9E/D
B1G, A21G, B9E, desB30
B1G,A21G,B9D,desB30 B1G.A21G, B10E/D
B1G, A21G, B10E, desB30 B1G, A21G, clesB25t desB30.
In another embodiment, the insulin is an analogue of human insulin from above three lists further modified in positions B3 and A18, eg B3T, B3S. B3Q and A18Q.
In another embodiment, the insulin is an analogue of human insulin from the above three lists further modified as follows: B3T, B28D B3T, desB27.
In another embodiment, the insulin is an analogue of human insulin from the above three lists further modified by deletion of B30. In another embodiment the ratio of the protamine-exteπded ligand of general for¬ mula (I) to zinc ion is 1:20 to 20:1.
In another embodiment the ratio of the protamiπe-extended ligand of general for¬ mula (I) to zinc ion is 1:6 to 10:1.
In another embodiment the amount of zinc ions is 2-6 moles per mole of putative in- sulin hexamer.
In another embodiment the amount of zinc ions is 2.0-3.5 moles per putative insulin hexamer.
In another embodiment zinc ions are present in an amount corresponding to 10 to 40 μg Zn/100 U insulin. In another embodiment zinc ions are present in an amount corresponding to 10 to 26 μg Zn/100 U insulin.
In another embodiment the ratio between insulin and the protamine-extended ligand of the invention is in the range from 99:1 to 1:99.
In another embodiment the ratio between insulin and the protamine-extended ligand of the invention is in the range from 95:5 to 5:95.
In another embodiment the ratio between between insulin and the protamine- extended ligand of the invention is in the range from 80:20 to 20:80.
In another embodiment the ratio between between insulin and the protamine- extended ligand of the invention is in the range from 70:30 to 30:70. In another aspect the invention relates to a method of preparing a protamine- extended ligand of the invention comprising the steps of:
• Identifying starter compounds that binds to the R-state HlsB10-Zn2* site
• optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids • attaching protamine. In another aspect the invention relates to a method of prolonging the action of an in¬ sulin preparation which comprises adding the ligand of the invention to the insulin prepara¬ tion.
In another aspect the invention relates to a method of treating type 1 or type 2 dia- betes comprising administering to a patient in need thereof a theraputicaHy effective amount of a pharmaceutical preparation comprising
• Insulin
• Zinc ions
• A ligand that binds to the R-state HisB10-Zn2* site, where said ligand may be as described in the embodiments above.
In another aspect the invention provides an embodiment 1, which is a pharmaceuti¬ cal preparation comprising
• Insulin • Zinc ions
• A ligand which binds reversibly to a HisB10 Zn2+ site of an R-state hexamer and wherein the ligand is extended by covalent attachment to protamine, having the following general formula (I)
CGr-Lnk-Frg-Protamine (I)
wherein:
CGr is a chemical group which reversibly binds to a His810 Zn2+ site of an insulin hexamer;
Lnk is a linker selected from
• a valence bond
• a chemical group GB of the formula -B1-B2-C(O)-, -B'-Ef-SOa-, -B1-B2-CH2-, or -B1- B2-NH-; wherein B1 is a valence bond. -O1 -S-, or -NR63-, B2 is a valence bond, C^de-alkylene, C2-C1θ-alkenylene, CrCie-alkynylene, arylene, heteroarylene, -CrC^-alkyl-aryl-, -Ca-C^-alkenyl-aryl-, -Ca-Ctβ-alkynyl-aryl-, -C(=O)-
Figure imgf000053_0001
alkyl-C(=O)-, -C(=O)- C,-C18-alkyl-S-C1-C1e-alky1-C(=O)-.
Figure imgf000053_0002
Ct8-alkyl-C(=0)-, -C(=O)-aryl-C(=O)-, -C(=O)-heteroaryl-C(=O}-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF3, -OCF3, -OR6B, or -NR65R78 and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR68, -C(O)H, OCORβB, -SO2, -CN, -CF3, -OCF3, -NO2. -0RβB. -NR6BR7B, CrC18-alkyl, or CrCi8-alkanoyl; R6Band R7B are independently H, CrC4-alkyl;
Frg is a fragment consisting of O to 5 neutral α- or β-amino acids, or
a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mix¬ ture of optical isomers, including a racemic mixture, or any tautomeric forms. Embodiment 2. A pharmaceutical preparation according to embodiment 1 wherein CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiophenolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano- 1 ,2,3-triazoles, benzimidazoles, benzotriazoles, purines, thiazolidinediones, tetrazoles, 5- mercaptotetrazoles, rhodanines. N-hydroxyazoles, hydantoines, thiohydantoines, barbitu¬ rates, naphthoic acids and salicylic acids. Embodiment 3. A pharmaceutical preparation according to embodiment 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2- πapthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or A- cyano-1 ,2,3-triazoles. Embodiment 4. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
Figure imgf000054_0001
wherein
X is =0, =S or =NH
Y is -S-, -O- or -NH-
R1, R1A and R4 are independently selected from hydrogen or Ci-C6-alkyl,
R2 and R2* are hydrogen or CrCe-alkyl or aryl, R1 and R2 may optionally be combined to form a double bond, R1A and R2* may optionally be combined to form a double bond,
R3, R3* and R5 are independently selected from hydrogen, halogen, aryl optionally substi- tuted with one or more substituents independently selected from R16. d-Ce-alkyl, or -C(O)NR11R12, A, A1 and B are independently selected from Ci-C6-alkyl, aryl, aryl-CrCfl-aIkyl, -NR11-ary[, aryI-C2-C6-alkenyl or heteroaryl, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from Re and the aryl or heteroaryl is optionally substituted with up to four substituents R7, R8, R9, and R10, A and R3 may be connected through one or two valence bonds, B and R5 may be connected through one or two valence bonds,
Rθ is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R7, R8, R9 and R10 are independently selected from
-hydrogen, halogen, -CN, -CH2CN1 -CHF2, -CF3, -OCF3. -OCHF2, -OCH2CF3,
-OCF2CHF2, -S(O)2CF3, -OS(O)2CF3. -SCF3, -NO2, -OR11, -NR11R12, -SR11, -NR11S(O)2R12, -S(O)2NR11R12, -S(O)NR11R12, -S(O)R11, -S(O)2R11, -OS(O)2 R1t, -C(O)NR11R12, -OC(O)NR11R12. -NR11C(O)R12, -CH2C(O)NR11R12,
-OCrCe-alkyl-C(O)NR11R12 r -CH2OR11, -CH2OC(O)R11, -CH2NR11R12, OC(O)R11, -OCrC15-alkyl-C(O)OR11, -OCrCβ-alkyl-OR11, -Sd-Ce-alkyl-qojOR11
-C2-Cβ-alkenyl-C(=O)0R11. -NR11-C(=O)-CrCe-alkyl-C(=0)0R11,
-NR11-C(=O)-C1-Cβ-alkenyl-C(=O)OR11 , -C(O)OR11, C(O)R11. or -C2-Cβ-alkenyl- C(=O)R11, =0, or -C2-Cβ-alkenyl-C(=O)-NR11R12,
•d-Cβ-alkyl, C2-Cβ-alkenyl or C2-Cβ-aIkynyI, each of which may optionally be substi¬ tuted with one or more.substituents independently selected from R13,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Ci-C8-alkoxy, aryl-CrCβ-alkyl, aryl-CrCe-alkenyl, aroyl-C2-C6-alkenyl. aryl-C^Ce-alkynyl, heteroaryl, heteroaryi-Cr Cβ-alkyl, heteroaryl-C2-C6-alkeπyl, heteroaryl-Cj-Ce-alkynyl, or C3-C6 cycloalkyl,
of which each cyclic moiety may optionally be substituted with one or more substitu¬ ents independently selected from R14,
R11 and R12 are independently selected from hydrogen, OH, Ci-C∞-alkyl, aryl-C^Ce-alkyl or aryl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substituents independently selected from R1β; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R13 is independently selected from halogen, -CN. -CF3, -OCF3, -OR11, -C(O)OR11 , -NR11R12, and -C(O)NR11R12,
R14 is independently selected from halogen, -C(O)OR11, -CH2C(O)OR11, -CH2OR11, -CN, - CF3, -OCF3, -NO2, -OR11, -NR11R12, -NR11C(O)R11. -S(O)2R11, aryl and CrCθ-alkyl,
R15 is independently selected from halogen. -CN, -CF3, =O, -OCF3, -OCrCβ-alkyl. -C(O)OCi- Cβ-alkyl, -COOH and -NH2,
R16 is independently selected from halogen, -C(O)OCrCβ-alkyl, -COOH, -CN. -CF3, -OCF3, - NO2. -OH, -OCt-Cβ-alkyl, -NH2, C(=0) or CrCβ-alkyl, or any enantiomer, diastereomer, in¬ cluding a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically accept¬ able acid or base. Embodiment 5. A pharmaceutical composition according to embodiment 4 wherein X is =O or =S.
Embodiment 6. A pharmaceutical composition according to embodiment 5 wherein X is =O. Embodiment 7. A pharmaceutical composition according to embodiment 5 wherein X is =S. Embodiment 8. A pharmaceutical composition according to any one of the embodiments 4 to 7 wherein Y is -O- or -S-.
Embodiment 9. A pharmaceutical composition according to embodiment 8 wherein Y is -O-.
Embodiment 10. A pharmaceutical composition according to embodiment 8 wherein Y is
-NH-.
Embodiment 11. A pharmaceutical composition according to embodiment 8 wherein Y is -S-. Embodiment 12. A pharmaceutical composition according to any one of the embodiments 4 to 11 wherein A is aryl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
Embodiment 13. A pharmaceutical composition according to embodiment 12 wherein A is selected from ArG1 optionally substituted with up to four substituents, R7, R8, RB, and R10 which may be the same or different.
Embodiment 14. A pharmaceutical composition according to embodiment 13 wherein A is phenyl or naphtyl optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different. Embodiment 15. A pharmaceutical composition according to embodiment 14 wherein A is
Figure imgf000057_0001
16. A pharmaceutical composition according to embodiment 14 wherein A is phenyl.
Embodiment 17. A pharmaceutical composition according to any one of the embodiments 4 to 11 wherein A is heteroaryl optionally substituted with up to four substitueπts, R7. R8, Rβ, and R10 which may be the same or different.
Embodiment 18. A pharmaceutical composition according to embodiment 17 wherein A is selected from Het1 optionally substituted with up to four substituents, Rτ, Rβ, Ra, and R10 which may be the same or different.
Embodiment 19. A pharmaceutical composition according to embodiment 18 wherein A is selected from Het2 optionally substituted with up to four substituents, R7 t R8, R9, and R10 which may be the same or different.
Embodiment 20. A pharmaceutical composition according to embodiment 19 wherein A is selected from Het3 optionally substituted with up to four substituents, R7, R8, R9, and Rt0 which may be the same or different.
Embodiment 21. A pharmaceutical composition according to embodiment 20 wherein A is selected from the group consisting of indolyl, benzofuranyl, quinolyl, furyl, thienyl, or pyrrolyl. wherein each heteroaryl may optionally substituted with up to four substituents, R7, R8, R9, and R10 which may be the same or different.
Embodiment 22. A pharmaceutical composition according to embodiment 20 wherein A is benzofuranyl optionally substituted with up to four substituents R7, R8. Rβ, and R10 which may be the same or different.
Embodiment 23. A pharmaceutical composition according to embodiment 22 wherein A is
Figure imgf000057_0002
24. A pharmaceutical composition according to embodiment 20 wherein A is carbazolyl op¬ tionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. Embodiment 25. A pharmaceutical composition according to embodiment 24 wherein A is
Figure imgf000058_0001
Embodiment 26. A pharmaceutical composition according to embodiment 20 wherein A is quinolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. Embodiment 27. A pharmaceutical composition according to embodiment 26 wherein A is
Figure imgf000058_0002
Embodiment 28. A pharmaceutical composition according to embodiment 20 wherein A is indolyl optionally substituted with up to four substituents R7, R8, R9, and R10 which may be the same or different. Embodiment 29. A pharmaceutical composition according to embodiment 28 wherein A is
Figure imgf000058_0003
Embodiment 30. A pharmaceutical composition according to any one of the embodiments 4 to 29 wherein R1 is hydrogen.
Embodiment 31. A pharmaceutical composition according to any one of the embodiments 4 to 30 wherein R2 is hydrogen.
Embodiment 32. A pharmaceutical composition according to any one of the embodiments 4 to 29 wherein R1 and R2 are combined to form a double bond.
Embodiment 33. A pharmaceutical composition according to any one of the embodiments 4 to 32 wherein R3 is d-Cβ-alkyl, halogen, or C(O)NR16R17. Embodiment 34. A pharmaceutical composition according to embodiment 33 wherein R3 is
C-Cβ-alkyl or C(O)NR18R17.
Embodiment 35. A pharmaceutical composition according to embodiment 34 wherein R3 is methyl. Embodiment 36. A pharmaceutical composition according to any one of the embodiments 4 to 11 wherein B is phenyl optionally substituted with up to four substituents, R7, Rs, R9, and R10 which may be the same or different.
Embodiment 37. A pharmaceutical composition according to any one of the embodiments 4 to 11 or 36 wherein R4 is hydrogen.
Embodiment 38. A pharmaceutical composition according to any one of the embodiments 4 to 11 or 36 to 37 wherein R5 is hydrogen.
Embodiment 39. A pharmaceutical composition according to any one of the embodiments 4 to 38 wherein R6 is aryl. Embodiment 40. A pharmaceutical composition according to embodiment 39 wherein Rβ is phenyl.
Embodiment 41. A pharmaceutical composition according to any one of the embodiments 4 to 40 wherein R7, Rβ, Rθ and R10 are independently selected from
•hydrogen, halogen. -NO2, -OR11, -NR11R12, -SR11, -NR11S(O)2R12, -S(O)2NR11R12,
-S(O)NR11R12, -S(O)R11, -S(O)2R11. -OS(O)2 R11. -NR11C(O)R12, -CH2OR11, - CH2OC(O)R11, -CH2NR11R12. -OC(O)R11. -OC1-C6^IKyI-C(O)OR11, -OC1-Cs- alkyl-C(O)NR11R12, -Od-Cβ-alkyl-OR11. -Sd-Cβ-alkyl-CtOJOR11, -C2-C6-alkenyl- C(O)OR11, -C(O)OR11. or-C2-Cβ-alkenyl-C(=O)R11,
• Ci-Cβ-alkyl, C2-Cβ-alkenyl or C2-Cβ-alkynyl, which may each optionally be substituted with one or more substituents independently selected from R13
•aryl, aryloxy, aroyl, arylsulfanyl, aryl-Ci-Cβ-alkoxy, aryl-CrCe-alkyl, aryl-C2- Cβ-alkenyl, aroyl-Cz-Ce-alkenyl, aryl-CrCβ-alkynyl, heteroaryl, heteroaryl-CrCa-alkyl, wherein each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
Embodiment 42. A pharmaceutical composition according to embodiment 41 wherein R7, Rβ, R9 and R10 are independently selected from
• hydrogen, halogen, -NO2, -OR11, -NR11R12, -SR11, -S(O)2R11, -OS(O)2 R11. - CH2OC(O)R11, -OC(O)R11, -Od-Cβ-alkyl-CtOJOR11, -OC-Cβ-alkyl-OR11, -SC1-C6- alkyl-C(O)OR11, -C(O)OR11. or-C2-Ce-alkenyl-C(=O)R11,
"Ci-Ce-alkyl or CrCδ-alkenyl which may each optionally be substituted with one or more substituents independently selected from R13 • aryl, aryloxy, aroyl, aryl-Ci-Ca-alkoxy, aryl-Ci-Ce-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
Embodiment 43. A pharmaceutical composition according to embodiment 42 wherein R7, R8, RB and R10 are independently selected from
• hydrogen, halogen, -NO2, -OR11, -NR11R12, -SR11, -S(O)2R11, -OS(O)2 R11. - CH2OC(O)R11, -OC(O)R11,
Figure imgf000060_0001
-SC1-C6- alkyl-C(O)OR11, -C(O)OR11, or-C2-Cβ-alkenyl-C(=O)R11,
•Ci-Cβ-alkyl or C1-Ce- which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aroyl, aryl-Ci-C6-alkoxy, aryl-CrCβ-alkyl, heteroaryl,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14. Embodiment 44. A pharmaceutical composition according to embodiment 43 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11, -OCi-Cβ-alkyF-C(O)OR11, or -C(O)OR11,
• CrC6-alkyl which may each optionally be substituted with one or more substituents independently selected from R13
• aryl, aryloxy, aryl-CrCβ-alkoxy,
of which each of the cyclic moieties optionally may be substituted with one or more substituents independently selected from R14.
Embodiment 45. A pharmaceutical composition according to embodiment 44 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen, -OR11, -OCrC6-alkyl-C(O)OR11, or -C(O)OR11,
•C1-C6-alkyl which may each optionally be substituted with one or more substituents independently selected from R13 •ArG1, ArG1oxyt ArG 1 -C1-C6-SIkOXy,
of which each of the cyclic moieties optionally may be substituted with one or more substitu- ents independently selected from R14.
Embodiment 46. A pharmaceutical composition according to embodiment 45 wherein R7, R8, R9 and R10 are independently selected from
• hydrogen, halogen. -OR11, -Od-Cβ-alkyl-CCOJOR11. or -C(O)OR",
•d-Cβ-alkyl which may optionally be substituted with one or more substituents inde¬ pendently selected from R13
• phenyl, phenyloxy, phenyl-Ci-C6-alkoxy, wherein each of the cyclic moieties option¬ ally may be substituted with one or more substituents independently selected from R14.
Embodiment 47. A pharmaceutical composition according to any one of the embodiments 4 to 46 wherein R11 and R12 are independently selected from hydrogen, CrC2o-alkyl, aryl or aryl-d-Cfi-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substi- tuted one or more substituents independently selected from R1β; R11 and R12 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitro¬ gen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds. Embodiment 48. A pharmaceutical composition according to embodiment 47 wherein R11 and R12 are independently selected from hydrogen, d-Czo-alkyl, aryl or aryl-C,-Cβ-alkyl, wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from R15, and the aryl groups may optionally be substituted one or more substitu¬ ents independently selected from R1Θ. Embodiment 49. A pharmaceutical composition according to embodiment 48 wherein R11 and R12 are independently selected from phenyl or phenyl-Ci-Cβ-alkyl.
Embodiment 50. A pharmaceutical composition according to embodiment 48 wherein one or both of R11 and R12 are methyl.
Embodiment 51. A pharmaceutical composition according to any one of the embodiments 4 to 50 wherein R13 is independently selected from halogen, CF3, OR11 or NR11R12. Embodiment 52. A pharmaceutical composition according to embodiment 51 wherein R13 is independently selected from halogen or OR11. Embodiment 53. A pharmaceutical composition according to embodiment 52 wherein R is
OR 11
Embodiment 54. A pharmaceutical composition according to any one of the embodiments 4 to 53 wherein R14 is independently selected from halogen, -C(O)OR11, -CN, -CF3, -OR11,
S(OJ2R", and d-Ce-alkyl.
Embodiment 55. A pharmaceutical composition according to embodiment 54 wherein R14 is independently selected from halogen, -C(O)OR11, or -OR11.
Embodiment 56. A pharmaceutical composition according to any one of the embodiments 4 to 55 wherein R15 is independently selected from halogen, -CN, -CF3, -CfOJOCrCβ-alkyl.and
-COOH.
Embodiment 57. A pharmaceutical composition according to embodiment 56 wherein R15 is independently selected from halogen or -C(O)OCi-Cβ-alkyl.
Embodiment 5θ. A pharmaceutical composition according to any one of the embodiments 4 to 57 wherein R1θ is independently selected from halogen, -C(O)OCrCa-alkyl, -COOH, -NO2,
-OCrCβ-alkyl, -NH2, C(=0) or d-Q-alkyl.
Embodiment 59. A pharmaceutical composition according to embodiment 58 wherein R19 is independently selected from halogen, -C(O)OCi-C6-afkyl, -COOH, -NO2, or d-C6-alkyl.
Embodiment 60. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
Figure imgf000062_0001
wherein
R1δ is hydrogen or CrCβ-alkyK
R20 is hydrogen or d-C6-alkyl,
D, D1 and F are a valence bond, d-Cβ-alkylene or d-C6-alkenylene optionally substituted with one or more substituents independently selected from R72,
R72 is independently selected from hydroxy. Ci-Ce-alkyl, or aryl, E is Ct-Cβ-alkyl, aryl or hβteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents R21, R22 and R23,
G and G1 are d-Cβ-alkyl, aryl or heteroaryl, wherein the aryl or heteroaryl is optionally sub¬ stituted with up to three substituents R24, R25 and R2β,
R17. R18. R21, R22, R23. R24, R25 and R26 are independently selected from
•hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -SCF3, -NO2, =0, -OR27, -NR27R28, -SR27, -NR27S(O)2R28, -S(O)2NR27R28, -S(O)NR27R28. -S(O)R27, -S(O)2R27, -C(O)NR27R28. -OC(O)NR27R26,
-NR27C(O)R28, -NR27C(O)OR28. -CH2C(O)NR27R2", -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28, -OC(O)R27, -OC-Cβ-alkyl-CfOJOR27. -SC1-C6-BlKyI-C(O)OR27, -C2-C6-
Figure imgf000063_0001
alkenyl-C(=O)OR27, -C(=O)NR27-CrCβ-alkyl-C(=0)0R27, -C1-C6-alkyl-C(=0)OR27,or -C(OPR27,
• CrCe-alkyl, C2-Cβ-alkenyl or C2-CB-alkynyl,
which may optionally be substituted with one or more substituents independently se- lected from R28,
• aryl. aryloxy. aryloxycarbonyl, aroyl, aryl-C,-Cβ-alkαxy, aryl-Ct-Cβ-alkyl, aryl-Cr Ce-alkenyl, aryl-C∑-Cβ-alkynyl, heteroaryl, heteroaryl-Ci-Cβ-alkyl, heteroaryl-C2-C9- alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30,
R27 and R28 are independently selected from hydrogen, Ci-C6-alkyl, aryl-CrCβ-alkyl or aryl, or R27 and R23 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R29 Is independently selected from halogen, -CN1 -CF3, -OCF3. -OR27, and -NR27R28, R30 is independently selected from halogen, -C(O)OR27, -CN, -CF3, -OCF3, -NO2, -OR27,
-NR27R28 and CrCβ-alkyl, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. Embodiment 61. A pharmaceutical composition according to embodiment 60 wherein D is a valence bond.
Embodiment 62. A pharmaceutical composition according to embodiment 60 wherein D is
C-pCe-alkylene optionally substituted with one or more hydroxy, C,-Ce-alkyl, or aryl.
Embodiment 63. A pharmaceutical composition according to any one of the embodiments 60 to 62 wherein E is aryl or heteroaryl, wherein the aryl or heteroaryl is optionally substituted with up to three substituents independently selected from R21, R22 and R23.
Embodiment 64. A pharmaceutical composition according to embodiment 63 wherein E is aryl optionally substituted with up to three substituents independently selected from R21, R22 and R23. Embodiment 65. A pharmaceutical composition according to embodiment 64 wherein E is selected from ArG1 and optionally substituted with up to three substttuents independently selected from R21, R22 and R23.
Embodiment 66. A pharmaceutical composition according to embodiment 65 wherein E is phenyl optionally substituted with up to three substituents independently selected from R21, R22 and R23.
Embodiment 67. A pharmaceutical composition according to embodiment 66 wherein CGr is
Figure imgf000064_0001
Embodiment 68. A pharmaceutical composition according to any one of the embodiments 60 to 67 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - NO2. -OR27, -NR27R28, -SR27, -C(O)NR27R28, -OC(O)NR27R28, -NR27C(O)R28, -NR27C(O)OR28, -CH2C(O)NR27R26. -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28, -OC(O)R27, -Od-C6-alkyl-C(O)OR27,
Figure imgf000064_0002
-Cs-Ce-al keπyl- C(=O)OR27, -NR27-C(=O)-CrC6-alkyl-C(=O)OR27, -NR^-Ct=O)-C1-Ce- alkeπyl-C(=O)OR27-, -C(=O)NR27-C1-Cβ-alkyl-C(=O)OR27, -C1-C6-alkyl-C(=O)OR27, or -C(O)OR27, •CrCe-alkyl, C2-Cβ-a!kenyl or CrCβ-alkynyl,
which may optionally be substituted with one or more substituents independently se- lected from R29
•aryl, aryloxy, aryloxycarbonyl. aroyl, aryl-Ci-Cβ-alkoxy, aryl-C-i-Cβ-alkyl, aryl-C2- Cβ-alkenyl, aryl-Ca-Cβ-alkynyl, heteroaryl, heteroaryl-Ci-C6-alkyl, heteroaryl-C2-Cβ- alkenyl or heteroaryl-Cz-Ce-alkyπyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
Embodiment 69. A pharmaceutical composition according to embodiment Error! Reference source not found, wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R23, -SR27, -NR27C(O)R28. -NR27C(O)OR28, -OC(O)R27, -OCrCe-alkyl-CfOpR27, -SCrCβ-alkyl-C(O)OR27, -C2-Cβ-alkenyl- C(=O)OR27. -C(=O)NR27-CrCθ-alkyI-C(=O)OR27, -C1-Cβ-alkyl-C(=O)OR27, or -C(O)OR27,
•Ci-Ce-alkyl optionally substituted with one or more substituents independently se¬ lected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-CrCβ-alkyl, heteroaryl, heteroaryl-CrC6- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
Embodiment 70. A pharmaceutical composition according to embodiment 69 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR26. -OC(O)R27, -OC,-CB-alkyl-C(O)OR27, -SCrCa-alkyl-C(O)OR27, -C;rC6-alkenyl- C(=O)OR27, -C(=0)NR27-C1-C6-alkyl-C(=O)0R27, -CrCβ-alkyl-C(=O)OR2T, or -C(O)OR27, •methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
•aryl, aryloxy, aroyl. aryl-d-Ce-alkoxy, aryl-CrC6-alkyl, heteroaryl, heteroaryf-d-Cβ- alkyl of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
Embodiment 71. A pharmaceutical composition according to embodiment 70 wherein R21, R22 and R23 are independently selected from
•hydrogen, halogen, -OCF3. -OR27, -NR27R23. -SR27. -NR27C(O)R23, -NR27C(O)OR28. -OC(O)R27, -Od-Cβ-alkyl-CtOJOR27, -SC,-C6-alkyl-C(O)OR27. -C2-Cε-alkenyl- C(=O)OR27, -C(=O)NR27-C1-Cβ-alkyl-C(=O)OR27. -C,-Cβ-alkyl-C(=O)OR27, or -C(O)OR27,
•methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
• ArG1, ArGI-O-, ArGI-C(O)-, ArG1-Ci-Cβ-alkoxy, ArGI-d-Cβ-alkyl, Het3, Het3-Cr Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30.
Embodiment 72. A pharmaceutical composition according to embodiment 71 wherein R2\ R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -OC1-Ca-alkyl-C(O)OR27, -SC, -CB-alky 1-C(O)OR27. -Ca-Ce-alkenyl- C(=O)OR27. -C(=O)NR27-C1-Cβ-alkyl-C(=O)OR27, -CrC6-alkyl-C(=O)OR27, or -C(O)OR27,
• d-Ce-alkyl optionally substituted with one or more substituents independently se¬ lected from R29
• phenyl, phenyloxy, phenyl-CrC6-alkoxy. phenyl-Ci-Ce-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30. Embodiment 73. A pharmaceutical composition according to any one of the embodiments 60 to 72 wherein R19 is hydrogen or methyl.
Embodiment 74. A pharmaceutical composition according to embodiment 73 wherein R1Θ is hydrogen. Embodiment 75. A pharmaceutical composition according to any one of the embodiments 60 to 74 wherein R27 is Hydrogen, Ci-Ce-alkyl or aryl.
Embodiment 76. A pharmaceutical composition according to embodiment 75 wherein R27 is hydrogen or CrCe-alkyl.
Embodiment 77. A pharmaceutical composition according to any one of the embodiments 60 to 76 wherein R28 is hydrogen or CrCβ-alkyl.
Embodiment 78. A pharmaceutical composition according to embodiment 60 wherein F is a valence bond.
Embodiment 79. A pharmaceutical composition according to embodiment 60 wherein F is Cr
Cβ-alkylene optionally substituted with one or more hydroxy, Ci-Cβ-alkyl, or aryl. Embodiment 80. A pharmaceutical composition according to any one of the embodiments 60 or 78 to 79 wherein G is Ci-Cβ-alkyl or aryl, wherein the aryl is optionally substituted with up to three substituents R24, R25 and R26.
Embodiment 81. A pharmaceutical composition according to any one of the embodiments 60 or 78 to 79 wherein G is CrCe-alkyl or ArGI1 wherein the aryl is optionally substituted with up to three substituents R24, R25 and R26.
Embodiment 82. A pharmaceutical composition according to embodiment 80 wherein G is
Ci-Cβ-alkyt.
Embodiment 83. A pharmaceutical composition according to embodiment 82 wherein G is phenyl optionally substituted with up to three substituents R24, R25 and R26. Embodiment 84. A pharmaceutical composition according to any one of the embodiments 60 to 83 wherein R24, R25 and R28 are independently selected from
• hydrogen, halogen, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -SCF3, - NO2, -OR27. -NR27R28, -SR27, -C(O)NR27R28, -OC(O)NR27R28, -NR27C(O)R28, -NR27C(O)OR28, -CH2C(O)NR27R28, -OCH2C(O)NR27R28, -CH2OR27, -CH2NR27R28.
-OC(O)R27, -OCrCβ-alkyl-CfOOR27, -SCrCβ-alkyl-C(O)OR27, -Ca-Cβ-alkenyl- C(=O)0R27, -NR27-C(=O)-C1-Cβ-alkyl-C(=O)OR27 t -NR27-C(=O)-Ci-C6- alkenyl-C(=O)OR27-, -C(=O)NR27-C1-Ce-alkyl-C(=O)OR27, -C1-C6-alkyl-C(=O)OR27, or -C(O)OR27,
• CrCβ-alkyl, C2-Cβ-alkenyl or Cz-Ce-alkynyl, which may optionally be substituted with one or more substituents independently se¬ lected from R29
•aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Ce-alkoxy, aryl-d-Ce-alkyl, aryl-Cr
Ce-alkenyl, aryl-C2-C6-a!kynyl. heteroaryl, heteroaryl-d-Oj-alkyl, heteroaryl-C2-Cβ- alkenyl or heteroaryl-Cz-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
Embodiment 85. A pharmaceutical composition according to embodiment 84 wherein R24, R25 and R28 are independently selected from
• hydrogen, halogen, -OCF3. -OR27, -NR27R28, -SR27, -NR27C(O)R28. -NR27C(O)OR28. -OC(O)R27, -OCrCβ-a[kyl-C(O)OR27, -Sd-Ce-alkyl-CfOJOR27, -CrC6-alkenyl-
C(=O)OR27, -C(=0)NR27-C1-Cβ-alkyl-C(=0)OR27, -C1-Cβ-alkyl-C(=O)OR27, or -C(O)OR27,
• d-Ce-alkyl, C2-C6-alkenyl or C2-Cβ-alkynyl,
which may optionally be substituted with one or more substituents independently se¬ lected from R29
• aryl, aryloxy, aryloxycarbonyl, aroyl, aryl-d-Ce-alkoxy, aryl-Ci-Cβ-alkyl, aryl-C2- C6-alkenyl, aryl-C2-Cβ-alkynyl, heteroaryl, heteroaryl-d-Ce-alkyl, heteroaryl-C2-Cβ- alkenyl or heteroaryl-CrCβ-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30. Embodiment 86. A pharmaceutical composition according to embodiment 85 wherein R24, R25 and R26 are independently selected from
• hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -Od-C6-alkyl-C(O)OR27, -Sd-Cβ-alkyl-C(O)OR27, -C2-C6-alkenyl- C(=O)OR27, -C(=O)NR27-CrCβ-alkyl-C(=O)OR27, -CrCβ-alkyl-C(=O)OR:!7, or
-C(O)OR27, •d-Ce-alkyl optionally substituted with one or more substitueπts independently se¬ lected from R29
»aryl, aryloxy, aroyl, aryl-Ci-Cβ-alkoxy, aryl-Ci-Cβ-alkyl, heteroaryl, heteroaryl-d-Ce- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30. Embodiment 87. A pharmaceutical composition according to embodiment 86 wherein R2\ R22 and R23 are independently selected from
•hydrogen, halogen, -OCF3, -OR27, -NR27R28. -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -OC,-Cβ-alkyl-C(O)OR27, -SCrCβ-alkyl-C(O)OR27. -C2-CB-alkenyl- Cf=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)OR27, -C1-C6-alkyl-C{=O)OR27, or
-C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
• ArG1, ArGI-O-, ArGI-C(O)-, ArGI-d-Cβ-alkoxy, ArG1-CrCβ-alkyI, Het3, Het3-Cr C6-alkyl of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R3D. Embodiment 88. A pharmaceutical composition according to embodiment 87 wherein R21, R22 and R23 are independently selected from
• hydrogen, halogen, -OCF3. -OR27, -NR27R28, -SR27, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -OC,-C8-alkyl-C(O)OR27, -SCrC3-alkyl-C(O)OR27, -CrC6-alkenyl- C(=O)OR27, -C(=O)NR27-CrC6-alkyl-C(=O)0R27, -Ci-C6-alkyl-C(=O)OR27, or
-C(O)OR27.
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29 •ArG1, ArGI-O-, ArGI-C(O)-, ArGi-C,-C6-a]koxy, ArG1-C,-C6-alkyl, Het3, Het3-C,- Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30. Embodiment 89. A pharmaceutical composition according to embodiment 88 wherein R21, R22 and R23 are independently selected from
•hydrogen, halogen, -OCF3, -OR27, -NR27R28, -SRΪ7, -NR27C(O)R28, -NR27C(O)OR28, -OC(O)R27, -OC-Cβ-alkyl-CtOJOR27, -SCrCβ-alkyl-C(O)OR27, -CrCβ-a1kenyl- C(=O)OR27, -C(=O)NR27-C1-Cβ-alkyl-C(=O)0R27, -C1-Cβ-a1kyl-C(=O)OR27, or
-C(O)OR27,
• methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R20
»ArG1, ArGI-O-, ArG1-CrCβ-alkoxy. ArGI-d-Cs-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30.
Embodiment 90. A pharmaceutical composition according to any one of the embodiments 60 or 78 to 89 wherein R20 is hydrogen or methyl. Embodiment 91. A pharmaceutical composition according to embodiment 90 wherein R20 is hydrogen/**
Embodiment 92. A pharmaceutical composition according to any one of the embodiments 60 or 78 to 91 wherein R27 is hydrogen, Ci-C6-alkyl or aryi.
Embodiment 93. A pharmaceutical composition according to embodiment 92 wherein R27 is hydrogen or Ci-Cβ-alkyl or ArG1.
Embodiment 94. A pharmaceutical composition according to embodiment 93 wherein R27 is hydrogen or CrCβ-alkyl.
Embodiment 95. A pharmaceutical composition according to any one of the embodiments 60 or 78 to 93 wherein R28 is hydrogen or d-Ce-alkyl. Embodiment 96. A pharmaceutical composition according to embodiment 60 wherein R17 and
R18 are independently selected from
• hydrogen, halogen, -CN1 -CF3, -OCF3, -NO2, -OR27, -NR27R28, -SR27, -S(O)R27, -S(O)2R27, -C(O)NR27R23, -CH2OR27, -OC(O)R27, -OC-Cβ-alkyl-CζOjOR27, -SC1-C6- alkyl-C(O)OR27, Or -C(O)OR27, *CrC6-alky!, Cz-Cβ-alkenyl or CrCθ-alkynyl, optionally substituted with one or more substituents independently selected from R2a
•aryl, aryloxy, aroyl, aryl-CrCβ-alkoxy, aryl-Ci-Cβ-alkyl, heteroaryl, heteroaryl-CrCe- alkyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
Embodiment 97. A pharmaceutical composition according to embodiment 96 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN. -CF3. -NO2, -OR27, -NR27R28. or -C(O)OR27,
•Ci-C6-alkyl optionally substituted with one or more substituents independently se- lected from R29
• aryl, aryloxy, aroyl, aryl-Ci-Cfl-alkoxy, aryl-CrCβ-alkyl, heteroaryl,
Figure imgf000071_0001
afkyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R30.
Embodiment 98. A pharmaceutical composition according to embodiment 97 wherein R17 and R1δare independently selected from
• hydrogen, halogen, -CN, -CF3, -NO2. -OR27. -NR27R28. or -C(O)OR27 • methyl, ethyl propyl optionally substituted with one or more substituents independ¬ ently selected from R29
• aryl, aryloxy, aroyl, aryl-d-Cβ-alkoxy, aryl-CrCe-alkyl, heteroaryl, heteroaryl-CrCβ- alkyl of which the cyclic moieties optionally may be substituted with one or more substitu- ents selected from R30.
Embodiment 99. A pharmaceutical composition according to embodiment 98 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN. -CF3, -NO2, -OR27, -NR27R28, or -C(O)OR27
• methyl, ethyl propyl optionally substituted with one or more substituents independ- ently selected from R29 •ArG1, ArGI-O-, ArGI-C(O)-, ArG1-CrC6-alkoxy, ArG1-CrCe-alkyl, Het3. Het3-C,- Ce-alkyl of which the cyclic moieties optionally may be substituted with one or more substitυents se¬ lected from R30. Embodiment 100. A pharmaceutical composition according to embodiment 99 wherein R17 and R18 are independently selected from
• hydrogen, halogen, -CN, -CF3, -NO2, -OR27, -NR27R28, or -C(O)OR27
• Ci-Cβ-alkyl optionally substituted with one or more substituents independently se¬ lected from R29 -phenyl, phenyloxy, phenyl-d-Cβ-aikoxy, phenyl-Ci-Cβ-alkyl, of which the cyclic moieties optionally may be substituted with one or more substituents se¬ lected from R30.
Embodiment 101. A pharmaceutical composition according to any one of the embodiments
60 to 100 wherein R27 is hydrogen or Ci-C6-alkyl. Embodiment 102. A pharmaceutical composition according to embodiment 101 wherein R27 is hydrogen, methyl or ethyl.
Embodiment 103. A pharmaceutical composition according to any one of the embodiments
60 to 102 wherein R2β is hydrogen or CrC6-alkyl.
Embodiment 104. A pharmaceutical composition according to embodiment 103 wherein R28 is hydrogen, methyl or ethyl.
Embodiment 105. A pharmaceutical composition according to any one of the embodiments
60 to 104 wherein R72 is -OH or phenyl.
Embodiment 106. A pharmaceutical composition according to embodiment 60 wherein CGr is
Figure imgf000072_0001
Embodiment 107. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is of the form H-I-J-
Figure imgf000072_0002
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R31
I is selected from
• a valence bond,
• -CH2N(R32J- Or -SO2N(R33)-.
•
Figure imgf000073_0001
wherein Z1 is S(O)2 or CH2, Z2 is -NH-, -O-or -S-, and π is 1 or 2,
J is
• Ci-Cβ-alkyl, C2-Cβ-alkeπyl or C2-Cβ-alkyπyl, which may each optionally be substituted with one or more substituents selected from R34,
•Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-CrCβ-alkoxy~, aryl-Ci-Cβ-alkyl-, aryl-C2- C6-alkenyl-, aryl-C2-Ce-alkynyl-, heteroaryl, heteroaryl-d-Cβ-alkyl-, heteroaryl-C2-C&- alkenyl- or
Figure imgf000073_0002
wherein the cyclic moieties are optionally substi¬ tuted with one or more substituents selected from R37, ■ hydrogen,
R31 is independently selected from hydrogen, halogen, SCN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -SCF3, -NO2. -OR35, -C(O)R35. -NR35R36, -SR35.
-NR35S(O)2R36, -S(O)2NR35R38, -S(O)NR35R36. -S(O)R35, -S(O)2R35, -C(O)NR35R39,
-OC(O)NR35R3a, -NR35C(O)R38. -CH2C(O)NR35R36, -OCH2C(O)NR35R38, -CH2OR35,
-CH2NR35R38, -OC(O)R35, -Od-Cβ-alkyl-CfOJOR35, -SCrCe-alkyl-CtopR35 _c2-Cβ-alkenyl-
C(=O)OR35, -NR3S-C(=O)-C1-C6-alkyl-C(=O)OR35 l -NR3S-C{=O)-CrC6-alkenyl-C(=0)0R35-, d-Cβ-alkyl, d-Ce-alkanoyl or -C(O)OR35,
R32 and R33 are independently selected from hydrogen, Ci-Cβ-alkyl or CrC6-alkanoyl,
R34 is independently selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R38,
R35 and R3β are independently selected from hydrogen, d-Cβ-alkyl. aryl-Ci-C6-alkyl or aryl, or R35 and R3β when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(O)OR33, -C(O)H, -CN, -CF3, -OCF3, -NO2. - OR35. -NR35R36, d-Cβ-alkyl or CrCe-alkanoyl,
or any eπantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 108. A pharmaceutical composition according to embodiment 107 wherein CGr is of the form H-I-J, wherein H is
Figure imgf000074_0001
wherein the phenyl, naphthalene or benzocarbazole rings are optionally substituted with one or more substituents independently selected from R31,
I is selected from *a valence bond,
• -CH2N(R32)- Or -SO2N(R33)-,
Figure imgf000074_0002
is S(O)2 or CH2, Z2 is N1-O-Or -S-, and n is 1 or 2,
J is • d-Cβ-alkyl, CjrCβ-alkenyl or Cz-Cβ-alkynyl, which may each optionally be substituted with one or more substituents selected from R34,
•Aryl, aryloxy, aryl-oxycarbonyl-, aroyl, aryl-Ci-Cβ-alkoxy-, aryt-Ci-Ce-alkyl-, aryl-C2- Ce-alkenyl-, aryl-Cz-Ce-alkynyl-, heteroaryl, heteroaryl-Ci-Ce-alkyl-, heteroaryl-C2-Ce- alkenyl- or heteroaryl-Cj-Cβ-alkynyh wherein the cyclic moieties are optionally substi- tuted with one or more substituents selected from R37,
• hydrogen, R31 is independently selected from hydrogen, halogen, -CN, -CH2CN1 -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -SCF3, -NO2, -OR35, -C(O)R35, -NR35R36, -SR35, -NR35S(O)2R38, -S(O)2NR35R38, -S(O)NR35R36. -S(O)R35, -S(O)2R35, -C(O)NR35R38, -OC(O)NR35R36, -NR35C(O)R36, -CH2C(O)NR35R38, -OCH2C(O)NR35R36, -CH2OR35, -CH2NR35R38, -OC(O)R35, -OC^Ce-alkyl-CfOpR36, -SCrQj-alkyl-CfOPR35 -Cz-Ce-alkenyl- C(=O)OR35, -NR3S-C(=O)-C1-Cθ-alkyl-C(=O)OR35, -NR35-C(=O)-C1-Ce-alkenyl-C(=O)OR33-. Ci-Cβ-alkyl. Ci-Cs-alkanoyl or -C(O)OR35,
R32 and R33 are independently selected from hydrogen. d-Cg-alkyl or CrCβ-alkanoyl,
R34Js independently selected from halogen, -CN, -CF3, -OCF3, -OR35, and -NR35R38,
R35 and R3β are independently selected from hydrogen, Ci-Cβ-alkyl, aryl-Ci-Cβ-alkyl or aryl. or R35 and R36 when attached to the same nitrogen atom together with the said nitrogen atom may form a 3 to 8 membered heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R37 is independently selected from halogen, -C(O)OR35, -C(O)H, -CN, -CF3, -OCF3, -NO2, - OR35, -NR35R38, d-Ce-alkyl or Ci-CB-alkanoyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base, With the proviso that R31 and J cannot both be hydrogen. Embodiment 109. A pharmaceutical composition according to any one of the embodiments 107 or 108 wherein H is
Figure imgf000075_0001
Embodiment 110. A pharmaceutical composition according to embodiment 109 wherein H is
Figure imgf000076_0001
Embodiment 111. A pharmaceutical composition according to embodiment 109 wherein H is
Figure imgf000076_0002
Embodiment 112. A pharmaceutical composition according to any one of the embodiments 107 to 111wherein I is a valence bond, -CH2N(R32)-, or SO2N(R33)-.
Embodiment 113. A pharmaceutical composition according to embodiment 112 wherein I is a valence bond.
Embodiment 114. A pharmaceutical composition according to any one of the embodiments
107 to 113 wherein J is •hydrogen,
• Ci-Ce-alkyl, Ca-Ce-alkenyl or C2-Cβ-alkynyl, which may optionally be substituted with one or more substituents selected from halogen. -CN, -CF3, -OCF3, -OR35, and -NR35R36,
• aryl, or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
Embodiment 115. A pharmaceutical composition according to embodiment 114 wherein J is
• hydrogen,
• aryi or heteroaryl, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37. Embodiment 116. A pharmaceutical composition according to embodiment 114 wherein J is
• hydrogen,
• ArG1 or Het3, wherein the cyclic moieties are optionally substituted with one or more substituents independently selected from R37.
Embodiment 117. A pharmaceutical composition according to embodiment 116 wherein J is • hydrogen,
• phenyl or naphthyl optionally substituted with one or more substituents inde¬ pendently selected from R37.
Embodiment 118. A pharmaceutical composition according to embodiment 117 wherein J is hydrogen. Embodiment 119. A pharmaceutical composition according to any one of the embodiments
107 to 118 wherein R32 and R33 are independently selected from hydrogen or d-CE-alkyl.
Embodiment 120. A pharmaceutical composition according to any one of the embodiments
107 to 119 wherein R34 is hydrogen, halogen, -CN, -CF3, -OCF3, -SCF3, -NO2, -OR35. -C(O)R35, -NR35R33, -SR35, -C(O)NR35R36, -OC(O)NR35R36, -NR35C(O)R36, -OC(O)R36, -OC1-
Cβ-alkyl-CpjOR35. -Sd-Cβ-alkyi-CtOJOR35 or -C(O)OR35.
Embodiment 121. A pharmaceutical composition according to embodiment 120 wherein R34 is hydrogen, halogen, -CF3. -NO2, -OR35, -NR35R36, -SR35, -NR35C(O)R38, Or-C(O)OR35.
Embodiment 122. A pharmaceutical composition according to embodiment 121 wherein R34 is hydrogen, halogen, -CF3, -NO2, -OR35, -NR35R38. or -NR35C(O)R36.
Embodiment 123. A pharmaceutical composition according to embodiment 122 wherein R34 is hydrogen, halogen, or -OR35.
Embodiment 124. A pharmaceutical composition according to any one of the embodiments
107 to 123 wherein R35 and R3β are independently selected from hydrogen, d-Cβ-alkyl, or aryl.
Embodiment 125. A pharmaceutical composition according to embodiment 124 wherein R35 and R39 are independently selected from hydrogen or Ci-Cβ-alkyl.
Embodiment 126. A pharmaceutical composition according to any one of the embodiments
107 to 125 wherein R37 is halogen, -C(O)OR35. -CN, -CF3, -OR33, -NR35R36, d-Cβ-alkyl or C1- C6-alkanoyl.
Embodiment 127. A pharmaceutical composition according to embodiment 126 wherein R37 is halogen, -C(O)OR35, -OR35, -NR35R36, CrCβ-alkyl or CrCβ-alkanoyl.
Embodiment 128. A pharmaceutical composition according to embodiment 127 wherein R37 is halogen, -C(O)OR35 or -OR35. Embodiment 129. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
Figure imgf000077_0001
wherein K is a valence bond, d-Cβ-atkylene, -NH-C(=O)-U-, -d-Cβ-alkyl-S-, -d-Cβ-afkyl-O-, -C(=O)-, or -C(=O)-NH-, wherein any d-Cβ-alkyl moiety is optionally substituted with R38,
U is a valence bond, d-Cβ-alkenylene, -d-C6-alkyl-O- or Ci-Cβ-alkylene wherein any C1- Ce-alkyl moiety is optionally substituted with Ci-Ce-alkyl, R38 is CrCβ-alky], aryl, wherein the alky] or aryl moieties are optionally substituted with one or more substituents independently selected from R3β,
R39 is independently selected from halogen, cyano, πitro, amino,
M is a valence bond, afylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R40,
R40 is selected from •hydrogen, halogen, -CN, -CH2CN, -CHF2, -CF3, -OCF3. -OCHF2, -OCH2CF3.
-OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3, -NO2, -OR41, -NR41R42, -SR41, -NR41S(O)2R42, -S(O)2NR41R42, -S(O)NR41R42. -S(O)R41, -S(O)2R41, -OS(O)2 R41, -C(O)NR41R42, -OC(O)NR41R42, -NR41C(O)R42, -CH2C(O)NR41R42, -OCrCβ- alkyl-C(O)NR41R42, -CH2OR41, -CH2OC(O)R41, -CH2NR41R42, -OC(O)R41, -OC,-Cfl- alkyl-C(O)OR4\ -Od-Cβ-alkyl-OR41, -S-d-Cβ-alkyl-CfOJOR41, -C2-C8-alkenyl-
Figure imgf000078_0001
alkenyl-C(=O)OR41 , -C(O)OR41, -C2-Cθ-alkenyl-C(=O)R41, =O, -NH-C^O)-O-C1- C9-alkyl, or -NH-C(=0)-C(=O)-O-C1-C6-alkyl,
•d-Ce-alkyl, Cj-Cβ-alkenyl or CrCβ-alkynyt, which may each optionally be substituted with one or more substituents selected from R43,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Ci-Cβ-alkoxy, aryl-d-Cβ-aIkyl, aryl-CrCβ-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-Cβ-alkynyl, heteroaryl, heteroaryl-d- Cβ-alkyl, heteroaryl-Cz-Cβ-alkenyl or heteroaryl-C2-Ce-alkynyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44,
R41 and R42 are independently selected from hydrogen, -OH1 Ci-Cθ-alkyl, CrC6-alkenyl, aryl- Ci-Cβ-afkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R45, and the aryl moieties may optionally be substi¬ tuted with one or more substituents independently selected from R46; R41 and R42 when at¬ tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds, R43 is independently selected from halogen, -CN. -CF3, -OCF3, -OR41, and -NR41R42
R44 is independently selected from halogen, -C(O)OR41, -CH2C(O)OR41, -CH2OR41, -CN, - CF3, -OCF3, -NO2, -OR41, -NR41R42 and CrC6-alkyl,
R45 is independently selected from halogen, -CN, -CF3, -OCF3, -O-d-Cβ-alkyl, -C(O)-O-C1-
Cβ-alkyl, -COOH and -NH2,
R46 is independently selected from halogen, -C(O)OCi-Cβ-alkyl, -COOH, -CN, -CF3, -OCF3, -
NO2, -OH, -OC-Cβ-alkyl, -NH2, C(=O) or d-Cβ-alkyl,
Q is a valence bond, d-Cβ-alkyleπe. -d-Ce-alkyl-O-, -d-Cβ-alkyl-NH-, -NH-C-Cβ-alkyl,
-NH-C(=O)-, -Cf=O)-NH-, -O-CrCβ-alkyl, -C(O)-, or -C1-Cβ-alkyl-C(=O)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48,
R47 and R4β are independently selected from hydrogen, d-Cβ-alkyl, aryl optionally substituted with one or more R49.
R49 is independently selected from halogen and -COOH,
T is
• hydrogen,
• Ci-Cβ-alkyl. C2-C6-alkenyl , C2-Cβ-alkynyl, d-Cβ-alkyloxy-carbonyl, wherein the alkyl, alkenyl and alkynyl moieties are optionally substituted with one or more substituents independently selected from R50,
• aryl, aryloxy, aryloxy-carbonyl, aryl-CrCβ-alkyl, aroyl, aryl-CrC6-alkoxy, aryl-Cz- Ce-alkenyl, aryl-CrC6-alkyny-, heteroaryl, heteroaryl-Ci-CB-alkyl, heteroaryl-C2- Ce-alkenylF heteroaryl-C2-Ca-alkynyl,
wherein any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R50,
R50 is d-Cβ-alkyl, CrCβ-alkoxy, aryl, aryloxy, aryl-Ci-C6-alkoxy, -C(=O)-NH-d-Ca-alkyl-aryl, -C(=O>NR50A-C1-C6-alky], -C(=O)-NH-(CH2CH20)mCi-C8-alkyl-COOH, heteroaryl, het- eroaryl-d-Ce-alkoxy, -d-Ce-alkyl-COOH, -O-Ci-Cβ-alkyl-COOH, -S(O)2R31, -Cz-Cβ-alkenyl-COOH, -OR51, -NO2, halogen. -COOH, -CF3, -CN, =O, -N(R51R52), wherein m is 1 , 2, 3 or 4, and wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53, and the alkyl moieties are optionally substituted with one or more R508. Embodiment R50* and R50B are independently selected from -C(O)OCrCβ-alkyl, -COOH, -C1- Cβ-alkyl-CtOOC-Ce-alkyl, -CrCβ-aikyUCOOH. or C,-Cβ-alkyl,
R51 and R52 are independently selected from hydrogen and d-Cβ-alkyl,
R53 Is independently selected from C-rCe-alkyl, d-Cβ-alkoxy, -d-Cβ-alkyl-COOH, -C2-
Cβ-alkenyl-COOH, -OR51, -NO2, halogen, -COOH, -CF3, -CN, Or -N(R51R52),
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 130. A pharmaceutical composition according to embodiment 129 wherein K is a valence bond, C,-Cβ-alkylene, -NH-C(=O)-U-, -d-Cβ-alkyl-S-, -d-Ce-alkyl-O-, or -C(=O)-, wherein any d-Cβ-alkyl moiety is optionally substituted with R38. Embodiment 131. A pharmaceutical composition according to embodiment 130 wherein K is a valence bond, d-CB-alkylene, -NH-C(=O)-U-, -Ci-Cβ-alkyl-S-, or -d-C6-alkyl-O, wherein any d-Cβ-alkyl moiety is optionally substituted with R38.
Embodiment 132. A pharmaceutical composition according to embodiment 131 wherein K is a valence bond, d-Ca-alkylene, or -NH-C(=O)-U, wherein any Ci-Cβ-alkyI moiety is optionally substituted with R38.
Embodiment 133. A pharmaceutical composition according to embodiment 132 wherein K is a valence bond or d-Ca-alkylene, wherein any d-Cβ-alkyl moiety is optionally substituted with R38.
Embodiment 134. A pharmaceutical composition according to embodiment 132 wherein K is a valence bond or -NH-C(=O)-U.
Embodiment 135. A pharmaceutical composition according to embodiment 133 wherein K is a valence bond.
Embodiment 136. A pharmaceutical composition according to any one of the embodiments
129 to 135 wherein U is a valence bond or -Ci-Cβ-alkyl-O-. Embodiment 137. A pharmaceutical composition according to embodiment 136 wherein U is a valence bond.
Embodiment 138. A pharmaceutical composition according to any one of the embodiments
129 to 137 wherein M is arylene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40. Embodiment 139. A pharmaceutical composition according to embodiment 138 wherein M is
ArG1 or Het1. wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
Embodiment 140. A pharmaceutical composition according to embodiment 139 wherein M is ArG1 or Het2, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
Embodiment 141. A pharmaceutical composition according to embodiment 140 wherein M is
ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40. Embodiment 142. A pharmaceutical composition according to embodiment 141 wherein M is phenylene optionally substituted with one or more substituents independently selected from
R40.
Embodiment 143. A pharmaceutical composition according to embodiment 141 wherein M is indolylene optionally substituted with one or more substituents independently selected from R40.
Embodiment 144. A pharmaceutical composition according to embodiment 143 wherein M is
Figure imgf000081_0001
145. A pharmaceutical composition according to embodiment 141 wherein M is carbazolylene optionally substituted with one or more substituents independently selected from R40.
Embodiment 146. A pharmaceutical composition according to embodiment 145 wherein M is
Figure imgf000081_0002
Embodiment 147. A pharmaceutical composition according to any one of the embodiments 129 to 146 wherein R40 is selected from -hydrogen, halogen, -CN, -CF3, -OCF3, -NO2, -OR41, -NR41R42, -SR41, -S(O)2R41.
-NR41C(O)R42, -OC,-Ce-alkyl-C(O)NR41R42, -C2-Cβ^lkenyl-C(=O)OR41, -C(O)OR41, =0, -NH-C^OJ-O-CrCe-alkyl, or -NH-C(=O)-C(=O)-O-CrCB-alkyl,
CrCβ-alkyl or C2-C8- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43, • aryl, aryloxy, ary]-Ci-Cβ-alkoxy, aryl-d-Cβ-alkyl, aryl-C^Ce-alkenyl, heteroaryl, het- eroaryl-CrCe-alkyl, or heteroaryt-C2-Ce-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substitueπts selected from R44.
Embodiment 148. A pharmaceutical composition according to embodiment 147 wherein R*0 is selected from
•hydrogen, halogen, -CN, -CF3, -OCF3, -NO2, -OR41, -NR41R42, -SR41, -S(O)2R41, -NR41C(O)R42, -OC,-Cβ-alkyl-C(O)NR41R42, -C2-Cθ-aIkenyl-C(=O)0R41, -C(O)OR41, =0, -NH-C(=O)-O-Ci-C6-alkyl1 or-NH-C(=0)-C(=0)-0-Ci-Cθ-alkyl,
d-Cβ-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• ArG1, ArGI-O-, ArGI-d-Cβ-alkoxy, ArG1-d-Cβ-alkyl. ArGI-Ca-Cβ-alkenyl, Het3, Het3-Ct-C6-alkyl, or Het3-C2-Cβ-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
Embodiment 149. A pharmaceutical composition according to embodiment 148 wherein R40 is selected from
• hydrogen, halogen. -CF3, -NO2. -OR41, -NR41R42, -C(O)OR41, =0. Or -NR41C(O)R42,
• d-Ce-alkyl, «ArG1.
Embodiment 150. A pharmaceutical composition according to embodiment 149 wherein R40 is hydrogen.
Embodiment 151. A pharmaceutical composition according to embodiment 149 wherein R40 is selected from • halogen, -NO2, -OR41, -NR41R42, -C(O)OR41, or -NR41C(O)R42,
• methyl,
• phenyl.
Embodiment 152. A pharmaceutical composition according to any one of the embodiments 129 to 151 wherein R41 and R42 are independently selected from hydrogen, d-Cβ-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
Embodiment 153. A pharmaceutical composition according to embodiment 152 wherein R41 and R42 are independently selected from hydrogen, methyl, ethyl, or phenyl, wherein the phenyl moieties may optionally be substituted with halogen or -COOH. Embodiment 154. A pharmaceutical composition according to any one of the embodiments 129 to 153 wherein Q is a valence bond, CrC6-alky[ene, -d-Ce-alkyl-O-, -Ci-Cβ-alkyl-NH-, -NH-d-Cβ-alkyl, -NH-C(=O)-, -C(=O)-NH-. -O-d-Cβ-alkyl, -C(=0)-, or -Ci- C6-alkyl-C{=O)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48.
Embodiment 155. A pharmaceutical composition according to embodiment 154 wherein Q is a valence bond, -CH2-. -CHrCH2-, -CH2-O-. -CH2-CHrO-, -CH2-NH-, -CH2-CHrNH-, -NH-CH2-, -NH-CH2-CHr, -NH-C(=O)-, -Ct=O)-NH-, -0-CH2-, -0-CH2-CHr, or -C(=O)-.
Embodiment 156. A pharmaceutical composition according to any one of the embodiments 129 to 155 wherein R47 and R48 are independently selected from hydrogen, methyl and phenyl.
Embodiment 157. A pharmaceutical composition according to any one of the embodiments 129 to 156 wherein T is
• hydrogen,
• Ci-C6-alkyl optionally substituted with one or more substituents independently se¬ lected from R50,
• aryl, aryl-d-Ce-alkyl, heteroaryl, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R50.
Embodiment 158. A pharmaceutical composition according to embodiment 157 wherein T is
• hydrogen,
• Ci-C6-alkyl optionally substituted with one or more substituents independently se¬ lected from R50, »ArG1, ArG1-Ci-Ce-atkyl, Het3, wherein the alkyl, aryl and heteroaryl moieties are op- tionally substituted with one or more substituents independently selected from R . Embodiment 159. A pharmaceutical composition according to embodiment 158 wherein T is
• hydrogen,
• d-Cβ-alkyl, optionally substituted with one or more substituents independently se- lected from R50,
• phenyl, phenyl-Ci-Cβ-alkyl, wherein the alkyl and phenyl moieties are optionally substituted with one or more substituents independently selected from R50.
Embodiment 160. A pharmaceutical composition according to embodiment 159 wherein T is phenyl substituted with R50. Embodiment 161. A pharmaceutical composition according to any one of the embodiments 129 to 160 wherein R50 is CrCβ*-alkyl, CrCβ-a!koxy, aryl, aryloxy, aryl-Ci-Ce-alkoxy, -C(=O)-NH-CrC6-alkyl-aryl, -C(=O)-NR50A-C1-Ce-alkyl, -C^Oi-NH-tCHaCHsOJmd-Cβ-alkyl- COOH1 heteroaryl, -d-Cø-alkyl-COOH, -O-d-Ce-alkyl-COOH, -S(O)2R51, -CrCβ-alkenyl-COOH, -OR51, -NO2, halogen. -COOH, -CF3, -CN. =0, -N(R51R52), wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53. Embodiment 162. A pharmaceutical composition according to embodiment 161 wherein R50 is d-Ce-alkyl, C,-Cβ-alkoxy, aryl, aryloxy, -C(=O)-NR50A-C1-Ce-alkyl1 -C^OVNH-fCHzCHzO CrCe-alkyl-COOH, aryl-C,-CB-alkoxy , -OR51, -NO2, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53. Embodiment 163. A pharmaceutical composition according to embodiment 162 wherein R50 is CrCβ-alkyl, aryloxy,
Figure imgf000084_0001
aryl-Ct-Cβ-alkoxy, -OR51, halogen, -COOH1 -CF3, wherein any aryl moiety is optionally substi¬ tuted with one or more R53. Embodiment 164. A pharmaceutical composition according to embodiment 163 wherein R50 is CrCβ-alkyl, ArGI-O-, -C(=O)-NR50A-CrCβ-alkyl, -C(=O)-NH-(CH2CH2O)mC,-C6-alkyl- COOH, ArG1-CrCβ-alkoxy , -OR51, halogen, -COOH, -CF3, wherein any aryl moiety is op¬ tionally substituted with one or more R53.
Embodiment 165. A pharmaceutical composition according to embodiment 164 wherein R50 is
Figure imgf000084_0002
Embodiment 166. A pharmaceutical composition according to embodiment 164 wherein R50 is phenyl, methyl or ethyl.
Embodiment 167. A pharmaceutical composition according to embodiment 166 wherein R50 is methyl or ethyl. Embodiment 168. A pharmaceutical composition according to any one of the embodiments 129 to 167 wherein m is 1 or 2.
Embodiment 169. A pharmaceutical composition accordϊhg to any one of the embodiments 129 to 168 wherein R51 is methyl.
Embodiment 170. A pharmaceutical composition according to any one of the embodiments 129 to 169 wherein R53 is Ci-Cβ-alkyl, Ci-C6-alkoxy, -OR51, halogen.or -CF3. Embodiment 171. A pharmaceutical composition according to any one of the embodiments 129 to 170 wherein R5oA is -C(O)OCH3, -C(O)OCH2CH3 -COOH, -CH2C(O)OCH3, - CH2C(O)OCH2CH3, -CH2CH2C(O)OCH3, -CH2CH2C(O)OCH2CH3, -CH2COOH1 methyl, or ethyl. Embodiment 172, A pharmaceutical composition according to any one of the embodiments 129 to 171 wherein RMB is -C(O)OCH3, -C(O)OCH2CH3 -COOH, -CH2C(O)OCH3, - CH2C(O)OCH2CH3, -CH2CH2C(O)OCH3, -CH2CH2C(O)OCH2CH3, -CH2COOH, methyl, or ethyl.
Embodiment 173. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is N=N
HN γH. "V-
wherein V is CrC6-alkyl, aryl, heteroaryl, aryl-C1-8-alkyl- or aryl-C2-β-alkenyl-, wherein the al- kyl or alkenyl is optionally substituted with one or more substituents independently selected from R54, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R5S,
R54 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2, R55 is independently selected from »hydrogen, halogen, -CN. -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3,
-OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3, -NO2, -OR56, -NR5$R57, -SR58, -NR56S(O)2R57, -S(O)2NR56R57, -S(O)NR56R57, -S(O)R58, -S(O)2R56, -OS(O)2 R58, -C(O)NR56R57, -OC(O)NR58R57, -NR56C(O)R57, -CH2C(O)NR56R57, -OC-C6- alkyl-CfOJNR^R57, -CH2OR56. -CH2OC(O)R56, -CH2NR56R57, -OC(O)R56, -OC1-C6- alkyl-CtOpR56, -OC-Cβ-alkyl-OR56, -SC-Ce-alkyl-CtOJOR56, -Cϋ-Cβ-alkenyl-
Cf=O)OR59, -NR5β-C(=O)-C1-Cβ-alkyl-C(=O)OR5θ, -NR5β-C(=0)-CrCβ- alkenyl-C(=O)OR5β , -C(O)OR56. or-C2-CB-alkenyl-C(=O)R56,
• Ci-Cβ-alkyl, Cr-Cβ-alkenyl or C2-C6-alkyny!,
which may optionally be substituted with one or more substituents selected from R5B,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Ci-C6-alkoxy, aryl-Ci-Cβ-alkyl, aryl-CrCe-alkenyl. aroyl-Cz-C6-alkenyl, aryl-Cz-Ce-alkynyl, heteroaryl, heteroaryl-Cr Cβ-alkyl, heteroaryl-C2-Ce-alkenyl or heteroaryl-C2-C8-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R5Θ.
R56 and R57 are independently selected from hydrogen, OH, CF3, d-C^-alkyl, aryl-CrCe- alkyl. -C(=O)-CrCa-alkyl or aryl. wherein the alkyl groups may optionally be substituted with one or more substituents independently selected from Rao, and the aryl groups may option¬ ally be substituted with one or more substituents independently selected from R61; R56 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further het- eroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R59 is independently selected from halogen, -CN, -CF3, -OCF3, -OR56, and -NR56R57,
R59 is independently selected from halogen. -C(O)OR56, -CH2C(O)OR56, -CH2OR56, -CN, - CF3, -OCF3, -NO2, -OR56, -NR56R57 and CrCβ-alkyl,
Rβ0 is independently selected from halogen, -CN, -CF3, -OCF3, -OCrCβ-alkylt -C(O)OCrC6- alkyl, -C(=O)-Re2 t -COOH and -NH2,
R61 is independently selected from halogen. -C(O)OCrCe-alkyl, -COOH. -CN1 -CF3, -OCF3, - NO2, -OH.
Figure imgf000086_0001
-NH2. C(=0) or C,-Ce-alkyl,
R62 is CrCβ-alkyl, aryl optionally substituted with one or more substituents independently se¬ lected from halogen, or heteroaryl optionally substituted with one or more
Figure imgf000086_0002
inde¬ pendently, or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
Embodiment 174. A pharmaceutical composition according to embodiment 173 wherein V is aryl, heteroaryl, or aryl-C1-8-alkyl-, wherein the alkyl is optionally substituted with one or more substituents independently selected R54, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R55. Embodiment 175. A pharmaceutical composition according to embodiment 174 wherein V is aryl, Het1 , or aryl-C^-alkyl-, wherein the alkyl is optionally substituted with one or more sub¬ stituents independently selected from R54, and the aryl or heteroaryl moiety is optionally sub¬ stituted with one or more substituents independently selected from R55. Embodiment 176. A pharmaceutical composition according to embodiment 175 wherein V is aryl, Het2, or aryl-C^-alkyl-, wherein the alkyl is optionally substituted with one or more sub¬ stituents independently selected from R54, and the aryl or heteroaryl moiety is optionally sub¬ stituted with one or more substituents independently selected from R55. Embodiment 177. A pharmaceutical composition according to embodiment 176 wherein V is aryl, Het3, or
Figure imgf000086_0003
wherein the alkyl is optionally substituted with one or more sub- stituents independently selected from R54. and the aryl or heteroaryl moiety is optionally sub¬ stituted with one or more substituents independently selected from Rss. Embodiment 178. A pharmaceutical composition according to embodiment 177 wherein V is aryl optionally substituted with one or more substituents independently selected from R55. Embodiment 179. A pharmaceutical composition according to embodiment 178 wherein V is ArG1 optionally substituted with one or more substituents independently selected from R55. Embodiment 180. A pharmaceutical composition according to embodiment 179 wherein V is phenyl, naphthyl or anthranyl optionally substituted with one or more substituents independ¬ ently selected from R55.
Embodiment 181. A pharmaceutical composition according to embodiment 180 wherein V is phenyl optionally substituted with one or more substituents independently selected from R55. Embodiment 182. A pharmaceutical composition according to any one of the embodiments 173 to 181 wherein R55 is independently selected from
• halogen, d-Cβ-alkyl, -CN1 -OCF3 ,-CF3, -NO2, -OR59, -NR56R57. -NR58C(O)R57 -SR58, -OCrCβ-alkyl-CtOOR58, or -C(O)OR58,
• Ci-Ce-alkyl optionally substituted with one or more substituents independently se- lected from R58
• aryl, aryl-d-Ce-alkyl, heteroaryl, or heteroaryl-d-Cβ-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents independently selected from R59.
Embodiment 183. A pharmaceutical composition according to embodiment 182 wherein R55 is independently selected from
• halogen, d-Ce-alkyl, -CN, -OCF3 ,-CF3, -NO2, -OR56,, -NR50R57, -NR56C(O)R57 -SR56, -OC1-C8-alkyl-C(O)OR5β, or -C(O)OR56
• Ci-Cβ-alkyl optionally substituted with one or more substituents independently se¬ lected from R58 • ArG1. AΓG1 -CrCβ-alkyl, Het3, or HetS-d-Ca-alkyl of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents independently selected from R59.
Embodiment 184. A pharmaceutical composition according to embodiment 183 wherein R55 is independently selected from halogen, -OR56, -NR56R57, -C(O)OR56, -OC1-C8- alkyl-CfOJOR56, -NR56C(O)R57 or CrCe-alkyl.
Embodiment 185. A pharmaceutical composition according to embodiment 184 wherein R55 is independently selected from halogen, -OR56, -NR56R57, -C(O)OR56, -OC1-Ca- alkyl-C(O)OR58, -NR56C(O)R57, methyl or ethyl. Embodiment 186. A pharmaceutical composition according to any one of the embodiments 173 to 185 wherein R58 and R57 are independently selected from hydrogen, CF3, Ci-Ci2-alkyI, or -C(=O)-C1-Ca-alkyl; R59 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
Embodiment 187. A pharmaceutical composition according to embodiment 186 wherein R56 and R57 are independently selected from hydrogen or CrCiralkyL R5β and R57 when at- tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
Embodiment 188. A pharmaceutical composition according to embodiment 187 wherein R56 and R57 are independently selected from hydrogen or methyl, ethyl, propyl butyl, R58 and R57 when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom.
Embodiment 189. A pharmaceutical composition according to any one of the embodiments 1 to 3 wherein CGr is
Figure imgf000088_0001
wherein AA is Ci-C6-alkyl, aryl, heteroaryl,
Figure imgf000088_0002
or aryl-C2-6-alkenyl-, wherein the alkyl or alkenyl is optionally substituted with one or more substituents independently selected from R63, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R64,
R63 is independently selected from halogen, -CN, -CF3, -OCF3, aryl, -COOH and -NH2.
R64 is independently selected from
• hydrogen, halogen. -CN, -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3,
-OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3. -NO2, -OR65, -NR65R66, -SR65, -NR85S(O)2R68. -S(O)2NR65R86, -S(O)NR65R68, -S(O)R65, -S(O)2R65. -OS(O)2 R65, -C(O)NR65R66, -OC(O)NR65R66, -NR65C(O)R66, -CH2C(O)NR65R66, -OC1-C6- 3^yI-C(O)NR65R66, -CH2OR65, -CH2OC(O)R65, -CH2NR65R66, -OC(O)R65, -OC1-Ca- alkyl-C(O)ORes. -Od-Cβ-alkyl-OR65, -SC1-Cδ-alkyl-C(O)OR6S, -C2-Cβ-alkenyl-
C(=O)OR65, -NR65-C(=O)-C1-Cβ-alkyl-C(=O)OR65, -NR65-C(=O)-C1-C6- alkenyl-C^OJOR65 , -C(O)OR65, or -C2-Cβ-alkenyl-C(=O)R65, • d-Ce-alkyl, CrCe-alkenyl or C2-Cβ-alkynylf each of which may optionally be substi¬ tuted with one or more substituents selected from Ra7,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-Ci-Cβ-alkoxy, aryl-CrCβ-alkyl, aryl-C2-Cβ-alkenyl.
Figure imgf000089_0001
aryl-CrCβ-alkynyl, heteroaryl, heteroaryl-Cr
Ce-alkyl, heteroaryl-CrCβ-alkenyl or heteroaryl-C2-C6-alkynyl,
of which the cyclic moieties optionally may be substituted with one or more substitu¬ ents selected from R68.
R85 and Rββ are independently selected from hydrogen, OH, CF3, Ci-Ci2-alkyl, aryl-CrCβ- alkyl, -C(=0)-Rβθ, aryl or heteroaryl, wherein the alkyl groups may optionally be substituted with one or more substituents selected from R70, and the aryl and heteroaryl groups may op¬ tionally be substituted with one or more substituents independently selected from R71; R85 and Rββ when attached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
Rβ7 is independently selected from halogen, -CN, -CF3, -OCF3. -OR85, and -NR65R68,
R6B is independently selected from halogen, -C(O)OR65, -CH2C(O)OR65, -CH2OR65, -CN, - CF3, -OCF3, -NO2, -OR65, -NR65R66 and Ci-Cβ-alkyl,
Rβθ is independently selected from Ct-Cβ-alkyl, aryl optionally substituted with one or more halogen, or heteroaryl optionally substituted with one or more Ci-Cβ-alkyl,
R70 is independently selected from halogen, -CN. -CF3, -OCF3, -OCrCβ-alkyl, -C(O)OC1-C6- alkyl, -COOH and -NH2,
R71 is independently selected from halogen, -C(O)OCi-Cθ-alkyl, -COOH, -CN, -CF3, -OCF3, - NO2, -OH, -OCrCβ-alkyl, -NH2. C(=0) or CrCβ-alkyl,
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base. Embodiment 190. A pharmaceutical composition according to embodiment 189 wherein AA is aryl, heteroaryl or aryl-C1-8-alkyl-, wherein the alkyl is optionally substituted with one or more R63, and the aryl or heteroaryl is optionally substituted with one or more substituents independently selected from R84. Embodiment 191. A pharmaceutical composition according to embodiment 190 wherein AA is aryl or heteroaryl optionally substituted with one or more substituents independently se¬ lected from Rθ4.
Embodiment 192. A pharmaceutical composition according to embodiment 191 wherein AA is ArG 1 or Het1 optionally substituted with one or more substituents independently selected from R64.
Embodiment 193. A pharmaceutical composition according to embodiment 192 wherein AA is ArG1 or Het2 optionally substituted with one or more substituents independently selected from R64. Embodiment 194. A pharmaceutical composition according to embodiment 193 wherein AA is ArG1 or Het3 optionally substituted with one or more substituents independently selected from R64.
Embodiment 195. A pharmaceutical composition according to embodiment 194 wherein AA is phenyl, naphtyl, anthryl, carbazolyl, thienyl, pyridyl, or benzodioxyl optionally substituted with one or more substituents independently selected from R84. Embodiment 196. A pharmaceutical composition according to embodiment 195 wherein AA is phenyl or naphtyl optionally substituted with one or more substituents independently se¬ lected from R64.
Embodiment 197. A pharmaceutical composition according to any one of the embodiments 189 to 196 wherein R84 is independently selected from hydrogen, halogen, -CF3, -OCF3, -OR65. -NR6SRββ, d-Ce-alkyl , -OC(O)R65, -OCi-C6-alkyl-C(O)ORe5. aryl-CrCe-alkeny1, ary- loxy or aryl, wherein CrCβ-alkyl is optionally substituted with one or more substituents inde¬ pendently selected from R67, and the cyclic moieties optionally are substituted with one or more substituents independently selected from R68. Embodiment 198. A pharmaceutical composition according to embodiment 197 wherein R64 is independently selected from halogen, -CF3, -OCF3, -OR65, -NR65R86, methyl, ethyl, propyl, -OC(O)R65, -OCHrC(O)OR65. -OCHrCH2-C(O)OR65, phenoxy optionally substituted with one or more substituents independently selected from R68.
Embodiment 199. A pharmaceutical composition according to any one of the embodiments 189 to 198 wherein R65 and Rβδ are independently selected from hydrogen, CF3, Ci-Cu-alkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently se¬ lected from R71. Embodiment 200. A pharmaceutical composition according to embodiment 199 wherein R85 and R88 are independently hydrogen, CrC12-aIkyl, aryl, or heteroaryl optionally substituted with one or more substituents independently selected from R71.
Embodiment 201. A pharmaceutical composition according to embodiment 200 wherein R65 and Rεβ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dϊmethyl-propyl, ArG1 or Het1 optionally substituted with one or more substituents independently selected from R71.
Embodiment 202. A pharmaceutical composition according to embodiment 201 wherein R65 and Rββ are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG 1 or Het2 optionally substituted with one or more substituents independently selected from R71. Embodiment 203. A pharmaceutical composition according to embodiment 202 wherein R65 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, ArG1 or Het3 optionally substituted with one or more substituents independently selected from R71.
Embodiment 204. A pharmaceutical composition according to embodiment 203 wherein Rβ5 and R66 are independently hydrogen, methyl, ethyl, propyl, butyl, 2,2-dimethyl-propyl, phenyl, naphtyl, thiadiazolyl optionally substituted with one or more R71 independently; or isoxazolyl optionally substituted with one or more substituents independently selected from R71.
Embodiment 205. A pharmaceutical composition according to any one of the embodiments
189 to 204 wherein R71 is halogen or d-Cβ-alkyl.
Embodiment 206. A pharmaceutical composition according to embodiment 205 wherein R71 is halogen or methyl.
Embodiment 207. A pharmaceutical preparation according to any one of the embodiments 1 to 205 wherein Frg consists of 0 to 5 neutral amino acids independently selected from the group consisting of GIy1 Ala, Thr, and Ser.
Embodiment 208. A pharmaceutical preparation according to embodiment 207 wherein Frg consists of 0 to 5 GIy.
Embodiment 209. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 0 GIy.
Embodiment 210. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 1 GIy. Embodiment 211. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 2 GIy.
Embodiment 212. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 3 GIy.
Embodiment 213. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 4 GIy. Embodiment 214. A pharmaceutical preparation according to embodiment 208 wherein Frg consists of 5 GIy.
Embodiment 215. A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein GB is of the formula B1-B2-C(O)-, B1-B2-SOz- or B1^-CHz- . wherein B1 and B2 are as defined in embodiment 1.
Embodiment 216. A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein GB is of the formula B1-B2-C(OK B1^-SO2- or B1-B2-NH-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 217. A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein GB is of the formula B'-B^CfO)-, B1^-CH2- or B1-B2-NH-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 218. A pharmaceutical preparation according to any one of the embodiments 1 to 214 wherein GB is of the formula B1^-CHr, B1-B2-Sθ2- or B1^-NH-, wherein B1 and B2 are as defined in embodiment 1. Embodiment 219. A pharmaceutical preparation according to any one of the embodiments
215 or 216 wherein GB is of the formula B'~B2-C(O)- or B1-B2-Sθ2-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 220. A pharmaceutical preparation according to any one of the embodiments
215 or 217 wherein GB is of the formula B1-B2-C(O)- or B1-B2-CHr, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 221. A pharmaceutical preparation according to any one of the embodiments
216 or 217 wherein GB is of the formula B'-B^CζO)- or B1^-NH-, wherein B1 and B2 are as defined in embodiment 1.
Embodiment 222. A pharmaceutical preparation according to any one of the embodiments 215 or 218 wherein GB is of the formula B1-B2-CH2- or
Figure imgf000092_0001
, wherein B1 and B2 are as defined in embodiment 1. Embodiment 223. A pharmaceutical preparation according to any one of the embodiments
216 or 218 wherein GB is of the formula B1-B2-NH- or B1^-SOr . wherein B1 and B2 are as defined in embodiment 1. Embodiment 224. A pharmaceutical preparation according to any one of the embodiments
217 or 218 wherein GB is of the formula B1-B2-CH2- or B1^-NH- , wherein B1 and B2 are as defined in embodiment 1.
Embodiment 225. A pharmaceutical preparation according to any one of the embodiments 219, 220, or 221 wherein GB is of the formula B1-B2-C(O)-. Embodiment 226. A pharmaceutical preparation according to any one of the embodiments 220, 222 or 224 wherein GB is of the formula B1-B2-CH;>-. Embodiment 227. A pharmaceutical preparation according to any one of the embodiments
220, 222 or 223 wherein GB is of the formula B1-B2-SO;r.
Embodiment 228. A pharmaceutical preparation according to any one of the embodiments
221 , 223 or 224 wherein GB is of the formula B'-BMMH-. Embodiment 229. A pharmaceutical preparation according to any one of the embodiments 1 to 228 wherein B1 is a valence bond, -O-, or -S-.
Embodiment 230. A pharmaceutical preparation according to any one of the embodiments 1 to 228 wherein B1 is a valence bond, -O-, or -N(R6B)-.
Embodiment 231. A pharmaceutical preparation according to any one of the embodiments 1 to 228 wherein B1 is a valence bond, -S-, or -N(R63)-.
Embodiment 232. A pharmaceutical preparation according to any one of the embodiments 1 to 228 wherein B1 is -O-, -S- or -N(R6V
Embodiment 233. A pharmaceutical preparation according to any one of the embodiments
229 or 230 wherein B1 is a valence bond or -O-. Embodiment 234. A pharmaceutical preparation according to any one of the embodiments
229 or 231 wherein B1 is a valence bond or -S-.
Embodiment 235. A pharmaceutical preparation according to any one of the embodiments
230 or 231 wherein B1 is a valence bond or -N(R6B)-.
Embodiment 236. A pharmaceutical preparation according to any one of the embodiments 229 or 232 wherein B1 is -O-or -S-.
Embodiment 237. A pharmaceutical preparation according to any one of the embodiments
230 or 232 wherein B1 is -O-or -N(RβB)-.
Embodiment 238. A pharmaceutical preparation according to any one of the embodiments
231 or 232 wherein B1 is -S-Or-N(R66)-. Embodiment 239. A pharmaceutical preparation according to any one of the embodiments 233, 234 or 235 wherein B1 is a valence bond. Embodiment 240. A pharmaceutical preparation according to any one of the embodiments
233. 236 or 237 wherein B1 is -O-.
Embodiment 241. A pharmaceutical preparation according to any one of the embodiments 234, 236 or 238 wherein B1 is -S-.
Embodiment 242. A pharmaceutical preparation according to any one of the embodiments
235. 237 or 238 wherein B1 is -N(R63)-.
Embodiment 243. A pharmaceutical preparation according to any one of the embodiments 1 to 242 wherein B2 is a valence bond, CrC18-alkylene, Cj-Ci β-alkenylene, Cz-Ci8-alkynylene, arylene, heteroarylene, -Cn-C^-alkyl-aryl-, -C(=O)-CrC1B-alkyl-C(=O)-, -C(=O)-CrC18-alkyl-
O-CrCie-alkyl-C(=O)-, -C(=O)-C1-C18-alkyl-S-C1-C1β-alkyl-C(=O)-, -C(=O)-Ci-Cia-alkyl-NR6- Ci-Cia-alkyl-C(=O)-; and the alkylene and arylene moieties are optionally substituted as de¬ fined in embodiment 1.
Embodiment 244. A pharmaceutical preparation according to embodiment 243 wherein B2 is a valence bond, d-Cia-alkylene, C2-C1a-alkenylene, Cz-C^-alkynylene, arylene, heteroary- lene, -d-C^-alkyl-aryl-, -C(=O)-C1-C1β-alkyl-C(=O)-, -C(=O)-CrCie-alkyl-O-Ci-C1e-alkyl- C(=O)-, and the alkylene and arylene moieties are optionally substituted as defined in em¬ bodiment 1.
Embodiment 245. A pharmaceutical preparation according to embodiment 244 wherein B2 is a valence bond, d-Cie-alkylene, C2-Ci8-alkenylene, CrCw-alkynylene, arylene, heteroary- lene, -d-Ciβ-alkyl-aryl-, -C(=O)-d-Ci8-alkyl-C(=O)-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
Embodiment 246. A pharmaceutical preparation according to embodiment 245 wherein B2 is a valence bond, d-dβ-alkylene, arylene, heteroarylene, -d-dβ-alkyl-aryl-, -C^O)-C1-C18- alkyl-C(=O)-1 and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
Embodiment 247. A pharmaceutical preparation according to embodiment 246 wherein B2 is a valence bond, d-Ci8-alkylene, arylene, heteroarylene, -d-de-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1. Embodiment 248. A pharmaceutical preparation according to embodiment 247 wherein B2 is a valence bond, Ci-Ci8-alkylene, arylene, -Ci-C18-alkyl-aryl-, and the alkylene and arylene moieties are optionally substituted as defined in embodiment 1.
Embodiment 249. A pharmaceutical preparation according to embodiment 248 wherein B2 is a valence bond or -CrC1B-alkylene, and the alkylene moieties are optionally substituted as defined in embodiment 1. Embodiment 250. A pharmaceutical preparation according to any one of the embodiments 1 to 249 wherein the insulin is selected from the group consisting of human insulin, an ana¬ logue thereof, a derivative thereof, and combinations of any of these. Embodiment 251. A pharmaceutical preparation according to embodiment 250 wherein the insulin is human insulin. Embodiment 252. A pharmaceutical preparation according to embodiment 250 wherein the insulin is an analogue of human insulin wherein position B28 is Asp, GIu, Lys, Leu, VaI or Ala.
Embodiment 253. A pharmaceutical preparation according to embodiment 252 wherein posi¬ tion B28 is Asp, GIu or Lys. Embodiment 254. A pharmaceutical preparation according to embodiment 253 wherein posi¬ tion B28 is Asp or GIu. Embodiment 255. A pharmaceutical preparation according to embodiment 254 wherein posi¬ tion B28 is Asp.
Embodiment 256. A pharmaceutical preparation according to embodiment 254 wherein posi¬ tion B28 is GIu. Embodiment 257. A pharmaceutical preparation according to any one of the embodiments 250 to 256 wherein the insulin is an analogue of human insulin wherein position 829 is Pro, Asp or GIu.
Embodiment 258. A pharmaceutical preparation according to embodiment 257 wherein posi¬ tion B29 is Pro or GIu. Embodiment 259. A pharmaceutical preparation according to embodiment 258 wherein posi¬ tion B29 is Pro.
Embodiment 260. A pharmaceutical preparation according to embodiment 258 wherein posi¬ tion B29 is GlU. Embodiment 261. A pharmaceutical preparation according to any one of the embodiments 250 to 260 wherein the insulin is an analogue of human insulin wherein position B9 is Asp or GIu.
Embodiment 262. A pharmaceutical preparation according to any one of the embodiments 250 to 261 wherein the insulin is an analogue of human insulin wherein position B10 is Asp or GIu. Embodiment 263. A pharmaceutical preparation according to embodiment 262 wherein posi¬ tion B10 is GIu.
Embodiment 264. A pharmaceutical preparation according to any one of the embodiments 250 to 263 wherein the insulin is an analogue of human insulin wherein position B1 is GIy. Embodiment 265. A pharmaceutical preparation according to any one of the embodiments 250 to 264 wherein the insulin is an analogue of human insulin wherein position B3 is Lys, Thr. Ser, Ala or GIn.
Embodiment 266. A pharmaceutical preparation according to embodiment 265 wherein posi¬ tion B3 is Lys, Thr, Ser or Ala. Embodiment 267. A pharmaceutical preparation according to embodiment 266 wherein posi- tion B3 is Lys or Ala.
Embodiment 268. A pharmaceutical preparation according to embodiment 267 wherein posi¬ tion B3 is Lys.
Embodiment 269. A pharmaceutical preparation according to any one of the embodiments 250 to 268 wherein the insulin is an analogue of human insulin wherein position B25 is de- leted. Embodiment 270. A pharmaceutical preparation according to any one of the embodiments 250 to 269 wherein the insulin is an analogue of human insulin wherein position B27 is de¬ leted.
Embodiment 271. A pharmaceutical preparation according to any one of the embodiments 250 to 270 wherein the insulin is an analogue of human insulin wherein position B30 is de¬ leted.
Embodiment 272. A pharmaceutical preparation according to any one of the embodiments 250 to 271 wherein the insulin is an analogue of human insulin wherein position A1Θ is GIn. Embodiment 273. A pharmaceutical preparation according to any one of the embodiments 250 to 272 wherein insulin is an analogue of human insulin wherein position A21 is Ala, GIn, GIu1 GIy, His, He, Leu, Met, Phe, Ser, Thr, Trp, Tyr, VaI or hSer.
Embodiment 274. A pharmaceutical preparation according to embodiment 273 wherein posi¬ tion A21 is Ala, GIy, lie, Leu, Phe, Ser, Thr, VaI or hSer. Embodiment 275. A pharmaceutical preparation according to embodiment 274 wherein posi- tion A21 is Ala or GIy.
Embodiment 276. A pharmaceutical preparation according to embodiment 275 wherein posi¬ tion A21 is GIy.
Embodiment 277. A pharmaceutical preparation according to any one of the embodiments 250 to 276 wherein the insulin is a derivative of human insulin or an analogue thereof having one or more lipophilic substituents.
Embodiment 278. A pharmaceutical preparation according to embodiment 277 wherein the Ne-amino group in position B29Lys is modified by covalent acylation with a hydrophobic moi¬ ety such as an fatty acid derivative or an litocholic acid derivative. Embodiment 279. A pharmaceutical preparation according to embodiment 277 or 278 wherein the insulin derivative is selected from the group consisting of B29-Ns-myristoyi- des(B30) human insulin, B29-Nε-palmitoyl-des(B30) human insulin, B29-Nε-myristoyl human insulin, B29-Nε-palmitoyl human insulin, B28-Nc-myristoyl LysB28 ProB29 human insulin, B28- Nε-palmitoyl LysB2e ProB2β human insulin, BSO-N^myristoyl-Thr^Lys830 human insulin, B30- Nε-palmitoyl-ThrB2βLysB30 human insulin, B29-NE-(N-palmitoyl-y-glutamyl)-des(B30) human insulin, B29-Nε-(N-lithocholyl-y-glutamyl>des(B30) human insulin, B29-Nε-(ω-carboxyhepta- decanoyl)-des(B30) human insulin and B29-Nε-(ω-carboxyheptadecanαyl) human insulin. Embodiment 280. A pharmaceutical preparation according to any one of the embodiments 250 to 279 wherein the insulin contain any combination of additional stabilizing substitutions. Embodiment 281. A pharmaceutical preparation according to embodiment 280 wherein the insulin contain any combination of the additional stabilizing substitutions in positions B1 , B3, A18 and A2i. Embodiment 282. A pharmaceutical preparation according to embodiment 250 wherein the insulin is an analogue of human insulin selected from the group:
B28D desB27 B28K.B29P
B3K.B29E
B29E desB25
B9E/D B10E/D.
Embodiment 283. A pharmaceutical preparation according to embodiment 250 wherein the insulin is an analogue of human insulin selected from the group:
A21G
A21G. B28K, B29P A21G. B28D
A21G, B28E
A21G, B3K, B29E
A21G, desB27
A21G, B9E A21G. B9D
A21G, B10E
A21G, desB25
A21G. desB30
A21G, B28K, B29P A21 G, B28K, B29P, desB30
A21G, B28D, desB30
A21G. B28E
A21G, B28E, desB30
A21G. B3K. B29E A21 G, B3K, B29E, desB30
A21G, desB27, desB30
A21G, B9E/D
A21G, B9E, desB30
A21G, B9D, desB30 A21G. B10E/D
A21G. B10E, desB30 A21G,desB25, desB30.
Embodiment 284. A pharmaceutical preparation according to embodiment 250 wherein the insulin is an analogue of human insulin selected from the group:
B1G.A21G B1G.A21G, B28K, B29P
B1G,A21G, B28D
B1G.A21G.B28E
B1G.A21G, B3K, B29E
B1G,A21G,desB27 B1G,A21GTB9E
B1G.A21G, B9D
B1G,A21G, B10E
B1G,A21G,desB25
B1G.A21G,desB30 B1G,A21G,B28K, B29P
B1G, A21G, B28K, B29P, desB30
B1G.A21G.B28D, desB30
B1G.A21G, B28E
B1G.A21G. B28E. desB30 B1G,A21G, B3K, B29E
B1 G1 A21 G1 B3K, B29E, desB30
B1G, A21G, desB27, desB30
B1G.A21G, B9E/D
B1G.A21G, B9E,desB30 B1 G, A21 G, B9D. desB30
B1G.A21G, B10E/D
B1G.A21G, B10E,desB30
B1G, A21G, desB25, desB30.
Embodiment 285. A pharmaceutical preparation according to any one of the embodiments 282 to 284 wherein the insulin is an analogue of human insulin further modified in positions
B3 and A18 as follows:
B3T
B3T.A18Q
B3S B3S, A18Q
B3Q B3Q. A18Q,
Embodiment 286. A pharmaceutical preparation according to any one of the embodiments 282 to 284 wherein the insulin is an analogue of human insulin further modified as follows: B3T, B28D B3T. desB27.
Embodiment 287. A pharmaceutical preparation according to to any one of the embodiments
282 to 286 wherein the insulin is an analogue of human insulin further modified by deletion of
B30.
Embodiment 288. A pharmaceutical preparation according to embodiments 1 to 287 wherein the ratio of the protamine-extended iigand of general formula (I) to zinc ion is 1 :20 to 20:1. Embodiment 289. A pharmaceutical preparation according to embodiment 288 wherein the ratio of the protamine-extended Iigand of general formula (I) to zinc ion is 1:6 to 10:1. Embodiment 290. A pharmaceutical preparation according to embodiments 1 to 289 wherein the amount of zinc ions is 2-6 moles per mole of putative insulin hexamer. Embodiment 291. A pharmaceutical preparation according to embodiment 290 wherein the amount of zinc ions is 2.0-3.5 moles per putative insulin hexamer.
Embodiment 292. A pharmaceutical preparation according to any one of the embodiments 1 to 291 wherein zinc ions are present in an amount corresponding to 10 to 40 μg Zn/100 U insu¬ lin. Embodiment 293. A pharmaceutical preparation according to embodiment 292 wherein zinc ions are present in an amount corresponding to 10 to 26 μg Zn/100 U insulin. Embodiment 294. A pharmaceutical preparation according to any one of the embodiments 1 to 293 wherein the ratio between insulin and the protamine-extended Iigand according to any one of the embodiments 1 to 249 is in the range from 99:1 to 1:99. Embodiment 295. A pharmaceutical preparation according to embodiment 294 wherein the ratio between insulin and the protamine-extended Iigand according to any one of the em¬ bodiments 1 to 249 is in the range from 95:5 to 5:95.
Embodiment 296, A pharmaceutical preparation according to embodiment 295 wherein the ratio between between insulin and the protamine-extended Iigand according to any one of the embodiments 1 to 249 is in the range from 80:20 to 20:80.
Embodiment 297. A pharmaceutical preparation according to embodiment 296 wherein the ratio between between insulin and the protamine-extended Iigand according to any one of the embodiments 1 to 249 is in the range from 70:30 to 30:70. Embodiment 298. A pharmaceutical preparation according to any one of the embodiments 1 to 297 wherein the concentration of insulin is 60 to 3000 πmol/ml. Embodiment 299. A pharmaceutical preparation according to embodiment 298 wherein the concentration of insulin is 240 to 1200 nmol/ml.
Embodiment 300. A pharmaceutical preparation according to embodiment 299 wherein the concentration of insulin is about 600 nmol/ml. Embodiment 301. A method of preparing a protamine-extended ligand according to embodi¬ ment 1 comprising the steps of
• Identifying starter compounds that binds to the R-state HisB10-Zn2+ site
• optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids
• attaching protamine. Embodiment 302. Method of prolonging the action of an insulin preparation which comprises adding a protamine-extended ligand according to any one of the embodiments 1 to 249 to the insulin preparation.
Embodiment 303. A method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation according to any one of the embodiments 1 to 300.
Embodiment 304. Use of a preparation according to any one of the embodiments 1 to 300 for the preparation of a medicament for treatment of type 1 or type 2 diabetes.
PHARMACEUTICAL PREPARATIONS
The present invention also relates to a pharmaceutical preparation for the treatment of diabetes in a patient in need of such a treatment comprising an R-state hexamer of insulin according to the invention together with a pharmaceutically acceptable carrier.
In one embodiment of the invention the insulin preparation comprises 60 to 3000 nmol/ml of insulin.
In another embodiment of the invention the insulin preparation comprises 300-2400 nmol/ml of insulin.
In another embodiment of the invention the insulin preparation comprises 240 to 1200 nmol/ml of insulin.
In another embodiment of the invention the insulin preparation comprises about 600 nmol/ml of insulin. Zinc ions may be present in an amount corresponding to 10 to 40 μg Zn/100 U insulin, more preferably 10 to 26 μg Zn/100 U insulin.
Insulin formulations of the invention are usually administered from multi-dose con¬ tainers where a preservative effect is desired. Since phenolic preservatives also stabilize the R-state hexamer the formulations may contain up to 50 mM of phenolic molecules. The phe- nolic molecules in the insulin formulation may be selected from the group consisting of phenol, m-cresol, chloro-cresol, thymol, 7-hydroxyindole or any mixture thereof.
In one embodiment the invention provides a pharmaceutical preparation further comprising at least 3 molecules of a phenolic compound per insulin hexamer. In another embodiment of the invention 0.5 to 4.0 mg/ml of phenolic compound may be employed.
In another embodiment of the invention 0.6 to 4.0 mg/ml of m-cresol may be em¬ ployed.
In another embodiment of the invention 0.5 to 4.0 mg/ml of phenol may be employed. In another embodiment of the invention 1.4 to 4.0 mg/ml of phenol may be employed.
In another embodiment of the invention 0.5 to 4.0 mg/ml of a mixture of m-cresol or phenol may be employed.
In another embodiment of the invention 1.4 to 4.0 mg/ml of a mixture of m-cresol or phenol may be employed. In another embodiment the invention provides a pharmaceutical preparation which may optionally contain a preservative such as e.g. phenol, m-cresol or mixtures thereof.
In another embodiment the invention provides a pharmaceutical preparation which may optionally contain an isotonicity agent such as e.g. NaCI, glycerol, mannitol and/or lac¬ tose. Chloride would be used at moderate concentrations (e.g. up to 50 mM) to avoid compe- tition with the zinc-site ligands of the present invention.
'* In another embodiment the the invention provides a pharmaceutical preparation which may optionally contain a buffer substance, such as a TRIS, phosphate, glycine or gly- cylglycine (or another zwitterionic substance) buffer
In another embodiment the the invention provides a pharmaceutical preparation which optionally comprises between 0.001 % by weight and 1 % by weight of a non-ionic sur¬ factant, for example tween 20 or Polox 188. A πonionic detergent can be added to stabilise insulin against fibrillation during storage and handling.
The action of insulin may further be slowed down in vivo by the addition of physio¬ logically acceptable agents that increase the viscosity of the pharmaceutical preparation. Thus, the pharmaceutical preparation according to the invention may furthermore comprise an agent which increases the viscosity, such as polyethylene glycol, polypropylene glycol, copolymers thereof, dextrans and/or polylactides.
In one embodiment the pharmaceutical preparation of the present invention may have a pH value in the range of 2.5 to 5.5, e.g. pH 2.5 to 4.5, pH 3 to 5.5. pH 3 to 4. In another embodiment insulin preparation of the present invention may have a pH value in the range of 3.5 to 8.5, e.g. pH 5.0 to 8.5, pH 5,5 to 8.5, pH 7.4 to 7.9. For pharmaceutical preparations of the present invention intended for formulation in the pH-rarige about 5.0-8.5, stabilizing mutations may include BIGIy1 des(B1), B3 may be Thr, Ser, or GIn, and A18 may be GIn.
For pharmaceutical preparations of the present invention intended for formulation in the pH-range 3.0-5.0 these substitutions may be combined with the A21 GIy stabilizing substi¬ tution.
In one embodiment the preparations of the invention are used in connection with in¬ sulin pumps. The insulin pumps may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable. Insulin pumps may be skin-mounted or carried, and the path of the insulin preparation from the storage compartment of the pump to the patient may be more or less tortuous. Non-limiting examples of insulin pumps are dis¬ closed in US 5.957,895, US 5.858,001, US 4,468,221 , US 4.468,221, US 5,957,895, US 5,858,001, US 6,074,369, US 5,858,001, US 5,527,288, and US 6,074,369.
In another embodiment the preparations of the invention are used in connection with pen-like injection devices, which may be prefilled and disposable, or the insulin preparations may be supplied from a reservoir which is removable. Non-limiting examples of pen-like in¬ jection devices are FlexPen®, innoLet®, InDuo™, Innovo®.
In a further embodiment preparations of the invention are used in connection with devices for pulmonary administration of aqueous insulin preparations, a non-limiting example of which is the AerX® device.
COMBINATION TREATMENT
The invention furthermore relates to treatment of a patient in which the pharmaceu¬ tical preparation of the invention, i.e. comprising zinc ions, insulin, eg human insulin, an ana¬ logue thereof, a derivative thereof or combinations of any of these analogue, acid-stabillised insulin, fast/rapid acting insulin and long/sfow/basal acting insulin, and a tigand for the R- state His810 Zn2+ site extended by protamine, is combined with another form of treatment.
In one aspect of the invention, treatment of a patient with the pharmaceutical prepa¬ ration of the invention is combined with diet and/or exercise.
In another aspect of the invention the pharmaceutical preparation of the invention is administered in combination with one or more further active substances in any suitable ratios. Such further active substances may e.g. be selected from antiobesity agents, antidiabetics, antihypertensive agents, agents for the treatment of complications resulting from or associ¬ ated with diabetes and agents for the treatment of complications and disorders resulting from or associated with obesity. Thus, in a further aspect of the invention the pharmaceutical preparation of the in¬ vention may be administered in combination with one or more antiobesity agents or appetite regulating agents.
Such agents may be selected from the group consisting of CART (cocaine am- phetamine regulated transcript) agonists, NPY (neuropeptide Y) antagonists, MC4 (melano- cortin 4) agonists, MC3 (melanocortin 3) agonists, orexin antagonists, TNF (tumor necrosis factor) agonists, CRF (corticotropin releasing factor) agonists, CRF BP (corticotropin releas¬ ing factor binding protein) antagonists, urocortin agonists, β3 adrenergic agonists such as CL-316243, AJ-9677, GW-0604, LY362884, LY377267 or AZ-40140, MSH (melanocyte- stimulating hormone) agonists, MCH (melanocyte-concentrating hormone) antagonists, CCK (cholecystokinin) agonists, serotonin re-uptake inhibitors such as fluoxetine, seroxat or cita- lopram, serotonin and noradrenaline re-uptake inhibitors, mixed serotonin and noradrenergic compounds, 5HT (serotonin) agonists, bombesin agonists, galanin antagonists, growth hor¬ mone, growth factors such as prolactin or placental lactogen, growth hormone releasing compounds, TRH (thyreotropin releasing hormone) agonists, UCP 2 or 3 (uncoupling protein 2 or 3) modulators, leptin agonists, DA agonists (bromocriptin, doprexin), lipase/amylase in¬ hibitors, PPAR (peroxisome proliferator-activated receptor) modulators, RXR (retinoid X re¬ ceptor) modulators, TR β agonists, AGRP (Agouti related protein) inhibitors, H3 histamine antagonists, opioid antagonists (such as naltrexone), exeπdin-4, GLP-1 and ciliary neurotro- phic factor.
In one embodiment of the invention the antiobesity agent is leptin. In another embodiment the antiobesity agent is dexamphetamiπe or amphetamine. In another embodiment the antiobesity agent is fenfluramine or dexfenfluramine. In still another embodiment the antiobesity agent is sibutramine. In a further embodiment the antiobesity agent is orlistat.
In another embodiment the antiobesity agent is mazindol or phentermine. In still another embodiment the antiobesity agent is phendimetrazine, diethylpropion, fluoxetine, bupropion, topiramate or ecopipam.
The orally active hypoglycemic agents comprise imidazolines, suiphonylureas, biguanides, meglitinides, oxadiazotidinediones, thiazolidinediones, insulin sensitizers, insulin secretagogues such as glimepride, α-giucosidase inhibitors, agents acting on the ATP- dependent potassium channel of the β-celis eg potassium channel openers such as those disclosed in WO 97/26265. WO 99/03861 and WO 00/37474 (Novo Nordisk A/S) which are incorporated herein by reference, or mitiglinide, or a potassium channel blocker, such as BTS-67582, nateglinide, glucagon antagonists such as those disclosed in WO 99/01423 and WO 00/39088 (Novo Nordisk A/S and Agouron Pharmaceuticals, Inc.), which are incorpo- rated herein by reference, GLP-1 agonists such as those disclosed in WO 00/42026 (Novo Nordisk A/S and Agouron Pharmaceuticals, Inc.), which are incorporated herein by refer¬ ence, DPP-IV (dipeptidyl peptidase-IV) inhibitors, PTPase (protein tyrosine phosphatase} in¬ hibitors, inhibitors of hepatic enzymes involved in stimulation of gluconeogenesis and/or gly- cogenolysis, glucose uptake modulators, GSK-3 (glycogen synthase kinase-3) inhibitors, compounds modifying the lipid metabolism such as antilipidemic agents, compounds lower¬ ing food intake, PPAR (peroxisome proliferator-activated receptor) and RXR (retinoid X re¬ ceptor) agonists, such as ALRT-268, LG-1268 or LG-1069.
In a further embodiment of the invention the pharmaceutical preparation of the in- vention is administered in combination with a sulphonylurea e.g. tolbutamide, chlorpropa¬ mide, tolazamide, glibenclamide, glipizide, glimepiride, glicazide or glyburide.
In another embodiment of the invention the pharmaceutical preparation of the inven¬ tion is administered in combination with a biguanide, e.g. metformin.
In yet another embodiment of the invention the pharmaceutical preparation of the in- vention is administered in combination with a meglitinide eg repaglinide or nateglinide.
In still another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with a thiazolidinedione insulin sensitizer, e.g. trogli- tazone, ciglitazone, pioglitazone, rosiglitazone, isaglitazone, darglitazone, englitazone, CS- 011/CI-1037 orT 174 or the compounds disclosed in WO 97/41097, WO 97/41119. WO 97/41120, WO 00/41121 and WO 98/45292 (Dr. Reddy's Research Foundation), which are incorporated herein by reference.
In still another embodiment of the invention the pharmaceutical preparation of the invention may be administered in combination with an insulin sensitizer, e.g. such as Gl 262570, YM-440, MCC-555, JTT-501 , AR-H039242, KRP-297, GW^09544, CRE-16336, AR-H049020, LY510929, MBX-102, CLX-0940, GW-501516 or the compounds disclosed in WO 99/19313, WO 00/50414, WO 00/63191, WO 00/63192, WO 00/63193 (Dr. Reddy's Re¬ search Foundation) and WO 00/23425, WO 00/23415, WO 00/23451 , WO 00/23445, WO 00/23417, WO 00/23416, WO 00/63153. WO 00/63196. WO 00/63209, WO 00/63190 and WO 00/63189 (Novo Nordisk A/S), which are incorporated herein by reference. In a further embodiment of the invention the pharmaceutical preparation of the in¬ vention is administered in combination with an α-glucosidase inhibitor, e.g. voglibose, emigli- tate, miglitol or acarbose.
In another embodiment of the invention the pharmaceutical preparation of the inven¬ tion is administered in combination with an agent acting on the ATP-dependent potassium channel of the β-cells, e.g. tolbutamide, glibenclamide, glipizide, glicazide, BTS-67582 or re¬ paglinide. In yet another embodiment of the invention the pharmaceutical preparation of the in¬ vention may be administered in combination with nateglinide.
In still another embodiment of the invention the pharmaceutical preparation of the invention is administered in combination with an antilipidemic agent, e.g. cholestyramine, colestipol, clofibrate, gemfibrozil, lovastatin, pravastatin, simvastatin, probucol or dextrothyroxine.
In another aspect of the invention, the pharmaceutical preparation of the invention is administered in combination with more than one of the above-mentioned compounds, e.g. in combination with metformin and a sulphonylurea such as glyburide; a sul phony lurea and acarbose; nateglinide and metformin; acarbose and metformin; a sulphonylurea, metformin and troglitazone; metformin and a sulphonylurea; etc.
Furthermore, the pharmaceutical preparation of the invention may be administered in combination with one or more antihypertensive agents. Examples of antihypertensive agents are β-biockers such as alprenolol, atenolol, timolol, pindolol, propranolol and metoprolol, ACE (angiotensin converting enzyme) inhibitors such as benazepril, captopril, enalapril, fosinopril, lisinopril, quinapril and ramipril, calcium channel blockers such as nifedipine, felodipine, nicardipine, isradipine, nimodipine, diltiazem and verapamil, and a- blockers such as doxazosin, urapidil, prazosin and terazosin. The pharmaceutical prepara¬ tion of the invention may also be combined with NEP inhibitors such as candoxatril. Further reference can be made to Remington: The Science and Practice of Phar¬ macy, 19th Edition, Gennaro, Ed., Mack Publishing Co., Easton, PA1 1995.
It should be understood that any suitable combination of the compounds according to the invention with diet and/or exercise, one or more of the above-mentioned compounds and optionally one or more other active substances are considered to be within the scope of the present invention.
EXAMPLES
The following examples and general procedures refer to intermediate compounds and final products identified in the specification and in the synthesis schemes. The prepara¬ tion of the compounds of the present invention is described in detail using the following ex- amples, but the chemical reactions described are disclosed in terms of their general applica¬ bility to the preparation of compounds of the invention. Occasionally, the reaction may not be applicable as described to each compound included within the disclosed scope of the inven¬ tion. The compounds for which this occurs will be readily recognised by those skilled in the art. In these cases the reactions can be successfully performed by conventional modiftca- tions known to those skilled in the art, that is, by appropriate protection of interfering groups, by changing to other conventional reagents, or by routine modification of reaction conditions. Alternatively, other reactions disclosed herein or otherwise conventional will be applicable to the preparation of the corresponding compounds of the invention. In all preparative methods, all starting materials are known or may easily be prepared from known starting materials. All temperatures are set forth in degrees Celsius and unfess otherwise indicated, all parts and percentages are by weight when referring to yields and all parts are by volume when refer¬ ring to solvents and eluents.
HPLC-MS (Method A)
The following instrumentation was used: • Hewlett Packard series 1100 G1312A Bin Pump
• Hewlett Packard series 1100 Column compartment
• Hewlett Packard series 1100 G13 15A DAD diode array detector
• Hewlett Packard series 1100 MSD
The instrument was controlled by HP Chemstation software.
The HPLC pump was connected to two eluent reservoirs containing:
A: 0.01 % TFA in water
B: 0.01% TFA in acetonitrile
The analysis was performed at 400C by injecting an appropriate volume of the sample (pref- erably 1 μL) onto the column, which was eluted with a gradient of acetonitrile.
The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table.
Figure imgf000106_0001
HPLC-MS (Method B) The following instrumentation was used:
Sciex AP1 100 Single quadropole mass spectrometer Perkin Elmer Series 200 Quard pump Perkln Elmer Series 200 autosampler Applied Biosystems 785A UV detector Sedex 55 evaporative light scattering detector
A Valco column switch with a Valco actuator controlled by timed events from the pump.
The Sciex Sample control software running on a Macintosh PowerPC 7200 computer was used for the instrument control and data acquisition.
The HPLC pump was connected to four eluent reservoirs containing:
A: Acetonitrife
B: Water
C: 0.5% TFA in water
D: 0.02 M ammonium acetate
The requirements for samples are that they contain approximately 500 μg/mL of the com¬ pound to be analysed in an acceptable solvent such as methanol, ethanol, acetonitrile, THF, water and mixtures thereof. (High concentrations of strongly eluting solvents will interfere with the chromatography at low acetonitrile concentrations.)
The analysis was performed at room temperature by injecting 20 μL of the sample solution on the column, which was eluted with a gradient of acetonitrile in either 0.05% TFA or 0.002 M ammonium acetate. Depending on the analysis method varying elutlon conditions were used.
The eluate from the column was passed through a flow splitting T-connector, which passed approximately 20 μUmin through approx. 1 m. 75 μ fused silica capillary to the API interface of AP1 100 spectrometer.
The remaining 1.48 mL/min was passed through the UV detector and to the E=LS detector.
During the LC-analysis the detection data were acquired concurrently from the mass spec¬ trometer, the UV detector and the ELS detector.
The LC conditions, detector settings and mass spectrometer settings used for the different methods are given in the following table. Column YMC ODS-A 120A s - 5μ 3 mm x 50 mm id
Gradient 5% - 90% acetonitrile in 0.05% TFA linearly during 7.5 min at 1.5 mL/min
Detection UV: 214 nm ELS: 40 0C
MS Experiment: Start: 100 amu Stop: 800 amu Step: 0.2 amu
Dwell: 0.571 msec
Method: Scan 284 times = 9.5 min
HPLC-MS (Method C) The following instrumentation is used:
• Hewlett Packard series 1100 G1312A Bin Pump
• Hewlett Packard series 1100 Column compartment
• Hewlett Packard series 1100 G1315A DAD diode array detector
• Hewlett Packard series 1100 MSD
• Sedere 75 Evaporative Light Scattering detector The instrument is controlled by HP Chemstation software.
The HPLC pump is connected to two eluent reservoirs containing:
0.01% TFA in water
B 0.01% TFA in acetonitrile
Thelpanalysis is performed at 40 0C by injecting an appropriate volumejof the sample (pref¬ erably 1 μ\) onto the column which is eluted with a gradient of acetonitrile. The HPLC conditions, detector settings and mass spectrometer settings used are given in the following table.
Figure imgf000108_0001
After the DAD the flow is divided yielding approximately 1 ml/min to the ELS and 0.5 ml/min to the MS. HPLC-MS (Method D) The following instrumentation was used: Sciex AP1 150 Single Quadropole mass spectrometer Hewlett Packard Series 1100 G1312A Bin pump Gilson 215 micro injector
Hewlett Packard Series 1100 G1315A DAD diode array detector Sedex 55 evaporative light scattering detector
A Valco column switch with a Valco actuator controlled by timed events from the pump. The Sciex Sample control software running on a Macintosh Power G3 computer was used for the instrument control and data acquisition.
The HPLC pump was connected to two eluent reservoirs containing: A: Acetonitrile containing 0.05% TFA
B: Water containing 0.05% TFA
The requirements for the samples are that they contain approximately 500 μg/ml of the com¬ pound to be analysed in an acceptable solvent such as methanol, ethanol. acetonitrile, THF1 water and mixtures thereof. (High concentrations of strongly eluting solvents will interfere with the chromatography at low acetonitrile concentrations.)
The analysis was performed at room temperature by injecting 20 μl of the sample solution on the column, which was eluted with a gradient of acetonitrile in 0.05% TFA The eluate from the column was passed through a flow splitting T-connector, which passed approximately 20 μl/min through approx. 1 m 75 μ fused silica capillary to the API interface of AP1 150 spectrometer.
The remaining 1.48 ml/min was passed through the UV detector and to the ELS detector. During the LC-analysis the detection data were acquired concurrently from the mass spec¬ trometer, the UV detector and the ELS detector. The LC conditions, detector settings and mass spectrometer settings used for the different methods are given in the following table.
Figure imgf000109_0001
EXAMPLES
Example 1 HBOL 1H-Benzotriazole
Figure imgf000110_0001
Example 2 HBOL 5,6-Dimethyl-i H-benzotriazole
Figure imgf000110_0002
Example 3 HBOL itf-Benzotriazole-5-carboxylic acid
Figure imgf000110_0003
Example 4 HBOL 4-Nitro-1H-benzotriazo!e
Figure imgf000110_0004
Example 5 HBOL 5-Amino-1 H-benzotriazole
Figure imgf000110_0005
Example 6 HBOL
5-Chlorσ-1 H-benzαtriazole
Figure imgf000111_0001
Example 7 HBOL
5-Nitro-1 H-benzotriazole
Figure imgf000111_0002
Example 8 PEM
4-[(1 H-Benzotriazole-5-carbonyl)amino]benzoic acid
Figure imgf000111_0003
4-[(1H-Benzotriazole-5-carbonyl)amino]benzoic acid methyf ester (5.2 g, 17.6 mmol) was dissolved in THF (60 mL) and methanol (10 mL) was added followed by 1N sodium hydrox¬ ide (35 mL). The mixture was stirred at room temperature for 16 hours and then 1N hydro¬ chloric acid (45 mL) was added. The mixture was added water (200 mL) and extracted with ethyl acetate (2 x 500 mL). The combined organic phases were evaporated in vacuo to afford 0.44 g of 4-[(1 H-benzotriazole-5-carbonyl)amino]benzoic acid. By filtration of the aqueous phase a further crop of 4-[(1H-benzotriazole-5-carbonyl)amino]benzoic acid was isolated (0.52 g).
1H-NMR (DMSO-dβ): δ 7.97 (4H, S)1 8.03 (2H1 m), 8.66 (1H, bs). 10.7 (1H, s), 12.6 (1H1 bs); HPLC-MS (Method A): m/z: 283 (M+1); Rt = 1.85 min. General procedure (A) for preparation of compounds of general formula I1:
Figure imgf000112_0001
wherein D, E and R19 are as defined above, and E is optionally substituted with up to three substituents R21, R22 and R23 independently as defined above.
The carboxylic acid of IH-benzotriazole-5-carboxylic acid is activated, ie the OH functionality is converted into a leaving group L (selected from eg fluorine, chlorine, bromine, iodine, 1- imidazolyl, 1,2,4-triazolyl, 1-benzotriazolyloxy, 1-(4-aza benzotriazolyl)oxy, pentafluoro- phenoxy, N-succinyloxy 3,4-dihydro-4-oxo-3-(1.2,3-benzotriazinyl)oxy. benzotriazole 5-COO. or any other leaving group known to act as a leaving group in acylation reactions. The acti¬ vated benzotriazote-5-carboxylic acid is then reacted with R2-(CH2)π-B' in the presence of a base. The base can be either absent (i.e. R2-(CH2)n-B' acts as a base) or triethylamine, N- ethyl-N,N.-diisopropylamine, N-methylmorpholiπe, 2,6-lutidine, 2,2,6,6-tetramethylpiperidine, potassium carbonate, sodium carbonate, caesium carbonate or any other base known to be useful in acylation reactions. The reaction is performed in a solvent solvent such as THF1 di- oxaπe, toluene, dichloromethane, DMF, NMP or a mixture of two or more of these. The reaction is performed between 0 0C and 800C, preferably between 20 0C and 400C. When the acylation is complete, the product is isolated by extraction, filtration, chromatography or other methods known to those skilled in the art
The general procedure (A) is further illustrated in the following example: Example 9 (General Procedure (A))PEM IH-Benzotriazole-5-carboxylic acid phenylamide
Figure imgf000112_0002
Benzotriazole-5-carboxylic acid (856 mg), HOAt (715 mg) and EDAC (1.00 g) were dissolved in DMF (17.5 mL) and the mixture was stirred at room temperature 1 hour. A 0.5 mL aliqot of this mixture was added to aniline (13.7 μL, 0.15 mmol) and the resulting mixture was vigor¬ ously shaken at room temperature for 16 hours. 1N hydrochloric acid (2 mL) and ethyl ace¬ tate (1 mL) were added and the mixture was vigorously shaken at room temperature for 2 hours. The organic phase was isolated and concentrated in vacuo to afford the title com¬ pound.
HPLC-MS (Method B): m/z: 239 (M+1); Rt = 3.93 min.
The compounds in the following examples were similarly made. Optionally, the compounds may be isolated by filtration or by chromatography.
Example 10 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (4-methoxyphenyl)amide
Figure imgf000113_0001
HPLC-MS (Method A): m/z: 269 (M+1) & 291 (M+23); Rt = 2.41 min HPLC-MS (Method B): m/z: 239 (M+1); Rt = 3.93 min.
Example 11 (General Procedure (A))PEM
{4-[(1 tt-Beπzorriazole-5-∞rbonyl)amino]phenyf}carbarπic add tert-butyl ester
Figure imgf000113_0002
HPLC-MS (Method B): m/z: 354 (M+1); Rt = 4.58 min.
Example 12 (General Procedure (A))PEM
IH-Benzotriazole-5-carboxylic acid (4-acetylaminophenyl)amide
Figure imgf000113_0003
HPLC-MS (Method B): m/z: 296 (M+1); Rt = 3.32 min.
Example 13 (General Procedure (A))PEM
IH-Benzotriazole-5-carboxylic acid (3-fluorophenyl)amide
Figure imgf000114_0001
HPLC-MS (Method B): m/z: 257 (M+1); Rt = 4.33 min.
Example 14 (General Procedure (A))PEM IH-Benzotrϊazole-5-carboxylic acid (2-chlorophenyl)amide
Figure imgf000114_0002
HPLC-MS (Method B): m/z: 273 (M+1); Rt = 4.18 min.
Example 15 (General Procedure (A))PEM 4-[(1 H-Benzotriazole-5-<arbonyl)amino]benzoic acid methyl ester
Figure imgf000114_0003
HPLC-MS (Method A):m/z: 297 (M+1); Rt : 2,60 min. HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.30 min.
Example 16 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (4-butylphenyl)amide
Figure imgf000114_0004
HPLC-MS (Method B): m/z: 295 (M+1); Rt = 5.80 min,
Example 17 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid (1 -phenylethyl)amide
Figure imgf000114_0005
HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.08 min. Example 18 (General Procedure (A))PEM IW-Benzotriazole-S-carboxylic acid beπzylamide
Figure imgf000115_0001
HPLC-MS (Method B): m/z: 253 (M+1); Rt = 3.88 min.
Example 19 (General Procedure (A))PEM IH-Benzotriazole-5-carboxylic acid 4~chlorobenzylamide
Figure imgf000115_0002
HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.40 min.
Example 20 (General Procedure (A))PEM IH-Benzαtriazole-5-carboxylic acid 2-chlorαbenzylamide
Figure imgf000115_0003
HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.25 min.
Example 21 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid 4-methoxybenzylamide
Figure imgf000115_0004
HPLC-MS (Method B): m/z: 283 (M+1); Rt = 3.93 min.
Example 22 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid 3-methoxybenzylamide
Figure imgf000115_0005
HPLC-MS (Method B): mSz: 283 (M+1); Rt = 3.97 min.
Example 23 (General Procedure (A))PEM
1 H-Benzotriazole-5-carboxylic acid (1,2-diphenylelhyl)amide
Figure imgf000116_0001
HPLC-MS (Method B): m/z: 343 (M+1); Rt = 5.05 min.
Example 24 (General Procedure (A))PEM 1tø-Benzotriazole~5-carboxylic acid 3-bromobenzylamide
Figure imgf000116_0002
HPLC-MS (Method B): m/z: 331 (M+1); Rt = 4.45 min.
Example 25 (General Procedure (A))PEM 4-{[(1 H-Benzotriazole-5-carbonyl)amino]methyl}benzoic acid
Figure imgf000116_0003
HPLC-MS (Method B): m/z: 297 (M+1 ); Rt = 3.35 min.
Example 26 (General Procedure (A))PEM 1 H-Beπzotriazote-5-carboxylic acid phenethylamide
Figure imgf000116_0004
HPLC-MS (Method B): m/z: 267 (M+1 ); Rt = 4.08 min.
Example 27 (Genera! Procedure (A))PEM IH-Benzotriazole-δ-carboxylic acid t2-(4-chloropheπyl)ethyl]amide
Figure imgf000116_0005
HPLC-MS (Method B): m/r. 301 (M+1); Rt = 4.50 min.
Example 28 (General Procedure (A))PEM 1H-Benzotriazole-5-carboxylic acid [2-(4-methoxyphenyl)ethyl]amide
Figure imgf000117_0001
HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.15 min.
Example 29 (General Procedure (A))PEM itf-Benzotriazole-δ-carboxylicacid [2-(3-methoxyphenyJ)ethyI]amide
Figure imgf000117_0002
HPLC-MS (Method B): m/z: 297 (M+1); Rt = 4.13 min.
Example 30 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic add [2-{3-chlorophenyI)ethyl]arnide
Figure imgf000117_0003
HPLC-MS (Method B): m/z: 301 (M+1); Rt = 4.55 min.
Example 31 (General Procedure (A))PEM 1 H-Benzotriazoϊe-5-carboχylic acid (2,2-diphenylethy1)amide
Figure imgf000117_0004
HPLC-MS (Method B): m/z: 343 (M+1); Rt = 5.00 min.
Example 32 (General Procedure (A))PEM 1 W-BenzotriazoIe-5-carboxylic acid (3,4-dichlorophenyl)methylamϊde
Figure imgf000117_0005
HPLC-MS (Method B): m/z: 321 (M+1); Rt = 4.67 min.
Example 33 (General Procedure (A))PEM itf-Benzotriazαle-5-carboxylic acid methytphenylamide
Figure imgf000118_0001
HPLC-MS (Method B): m/z: 253 (M+1); Rt = 3.82 min.
Example 34 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid benzylmethylamide
Figure imgf000118_0002
HPLC-MS (Method B): m/z: 267 (M+1); Rt = 4.05 min.
Example 35 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid [2-(3-chloro-4-methoxyphenyl)ethyllmethyl*amide
Figure imgf000118_0003
HPLC-MS (Method B): m/z: 345 (M+1); Rt = 4.37 min.
Example 36 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxylic acid rήethylphenethylamide
Figure imgf000118_0004
HPLC-MS (Method B): m/z: 281 (M+1); Rt = 4.15 min.
Example 37 (General Procedure (A))PEM IH-Benzotriazole-5-carboxylic acid [2-(3,4-dimethoxyphenyl)ethyi]methylamide
Figure imgf000118_0005
HPLC-MS (Method B): m/z: 341 (M+1); Rt = 3.78 min;
Example 38 (General Procedure (A))PEM 1 H-Benzotriazole-5-carboxyJic acid (2-hydroxy-2-phenylethyl)methylamide
Figure imgf000119_0001
HPLC-MS (Method B): m/z: 297 (M+1); Rt = 3.48 miπ.
Example 39 (General procedure (A)) IH-Benzotriazole-S-carboxylic acid (3-bromophenyl)amide
Figure imgf000119_0002
HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.19 min.
Example 40 (General procedure (A)) 1H-Benzotriazole-5-carboxylic acid (4-bromophenyl)amide
Figure imgf000119_0003
HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.18 min.
■y
Example 41 (General procedure (A)) {4-[(1 H-Benzotriazole-5-carbonyl)amino]benzoylamino}acetic acid
Figure imgf000119_0004
HPLC-MS (Method A): m/z: 340 (M+1); Rt = 1.71 min.
Example 42 (General procedure (A)) {4-[(1H-Benzotriazole-5-carbonyl)amino]phenyl}acetic acid
Figure imgf000119_0005
HPLC-MS (Method A): m/z: 297 (M+1); Rt = 2.02 min. Example 43 (General procedure (A))
3-{4-[(1 H-Benzotriazole-δ-carbonyOaminolphenytyacrylic acid
Figure imgf000120_0001
HPLC-MS {Method A): m/z: 309 (M+1); Rt = 3.19 min.
Example 44 (General procedure (A))
{3-[(1 H-Benzotriazole-5-rarboπyl)amino]phenyl}acetic acid
Figure imgf000120_0002
HPLC-MS (Method A): m/z: 297 (M+1 ); Rt a 2.10 min.
Example 45 (General procedure (A)) 2-{4-[{1H-Benzotriazole-5-carbonyl)amiπo]phenoxy}-2-methylpropioπic acid
Figure imgf000120_0003
HPLC-MS (Method A): m/z: 341 (M+1 ); Rt = 2.42 min.
Example 46 (General procedure (A)) S-^-KIH-Benzotriazole-δ-carbonytyamiπolbenzoylaminoJpropionic acid
Figure imgf000120_0004
HPLC-MS (Method A): m/z: 354 (M+1 ); Rt = 1.78 min.
Example 47 (General procedure (A))
3-(4-[(I H-Benzotriazole-5-carbonyl)amino]phenyl}propionic acid
Figure imgf000121_0001
HPLC-MS (Method A): m/z: 311 (M+1); Rt a 2.20 min.
Example 48 (General procedure (A)) 1 H-BenzotriazoIe-5-carboxylic acid (4-benzyfoxypheπyl)amide
Figure imgf000121_0002
HPLC-MS (Method A): m/z: 345 (M+1); Rt = 3.60 min.
Example 49 (General procedure (A)) 1 H'Benzotriazole-δ-carboxylic acid (3-chloro-4-methoxyphenyl)amide
Figure imgf000121_0003
* HPLC-MS (Method A): m/z: 303 (M+1 ); Rt = 2.88 min.
Example 50 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (4-phenoxyphenyl)amide
Figure imgf000121_0004
HPLC-MS (Method A): m/z: 331 (M+1); Rt = 3.62 min.
Example 51 (General procedure (A)) 1 H-Benzotriazole-5-carboxylic acid (4-butoxyphenyl)amide
HPLC-MS (Method A): m/z: 311 (M+1); Rt = 3.59 min.
Example 52 (General procedure (A))
1 H-Benzotriazole-S-carboxylic acid (3-bromo-4-trifluoromethoxyphenyl)amide
Figure imgf000122_0001
HPLC-MS (Method A): m/z: 402 (M+1); Rt = 3.93 min.
Example 53 (General procedure (A))
IH-Benzotriazoie-5-carboxyiic acid {3,5-dichloro-4-hydroxyphenyl)amide
Figure imgf000122_0002
HPLC-MS (Method A): m/z: 323 (M+1); Rt = 2.57 min.
Example 54 (General procedure (A))
4-{[(1 H-Benzotriazole-5-carbonyl)amiπo]methyl}benzoic acid
Figure imgf000122_0003
HPLC-MS (Method A): m/z: 297 (M+1); Rt = 1.86 min.
Example 55 (General procedure (A))
{4-[{1 H-Benzotriazole-S-carbonylJaminolphenylsulfany^acetic acid
Figure imgf000122_0004
HPLC-MS (Method A): m/z: 329 (M+1); Rt = 2.34 min. Example 56
N-(1 H-Benzotriazol-5-yl)acetamide
Figure imgf000123_0001
HPLC-MS (Method A): m/z: 177 (M+1); Rt = 0.84 min.
Example 57 (General Procedure (A)) itf-Benzotriazole-S-carboxylic acid 4-nitrobenzylamide
Figure imgf000123_0002
The following compound Is prepared according to general procedure (N) as described below: Example 58 (General procedure (N)) IH-Benzotriazole-5-carboxylic acid 4-chlorobenzylamide
Figure imgf000123_0003
HPLC-MS (Method B): m/z: 287 (M+1); Rt = 4.40 min.
Example 59 2-[(1H-Benzotriazol-5-y!imino)methyl]-4,6-dichlorophenol
Figure imgf000123_0004
Example 60 Diethyl 2-[(1 H-benzotriazol-6-ylamino)methylidene]malonate
Figure imgf000124_0001
Example 61 N1 -(1 H-Benzotriazol-5-yl>3-chlorαbenzamide
Figure imgf000124_0002
Example 62 N 1 -{ 1 H-Benzotriazof-5-y!>3 ,4,5-trimetho>ybenzamide
Figure imgf000124_0003
Example 63 N2-<1 H-BenzotriazoI-5-yl)-3-ch!orobenzo[b]thiophene-2-carboxamide
Figure imgf000124_0004
Example 64 6-Bromo-1 H-benzotriazole
Figure imgf000125_0001
Example 65 2-[(1 H-Benzotriazol-5-ylimino)methyl]-4-bromophenol
Figure imgf000125_0002
General procedure (B) for preparation of compounds of general formula I2
Figure imgf000125_0003
U wherein X, Y, A and R3 are as defined above and A is optionally substituted with up to four substitueπts R7, Ra, R8, and R10 as defined above.
The chemistry is well known (eg Lohray et a!., J. Med. Chem., 1999, 42, 2569-81) and is generally performed by reacting a carbonyl compound (aldehyde or ketone) with the hetero¬ cyclic ring (eg thiazolidiπe-2,4-dione (X = O; Y = S)1 rhodanine (X = Y = S) and hydantoin (X = O; Y = NH) in the presence of a base, such as sodium acetate, potassium acetate, ammo¬ nium acetate, piperidinium beπzoate or an amine (eg piperidine, triethylamine and the like) in a solvent (eg acetic acid, ethanol, methanol, DMSO, DMF, NMP1 toluene, benzene) or in a mixture of two or more of these solvents. The reaction is performed at room temperature or at elevated temperature, most often at or near the boiling point of the mixture. Optionally, azeotropic removal of the formed water can be done.
This general procedure (B) is further illustrated in the following example: Example 66 (General procedure (B)) 5-(3-Phenoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000126_0001
A solution of thiazolidine-2,4-dione (90%, 78 mg, 0.6 mmol) and ammonium acetate (92 mg, 1.2 mmol) in acetic acid (1 mL) was added to 3-phenoxybeπzaldehyde (52 μL, 0.6 mmol) and the resulting mixture was shaken at 1150C for 16 hours. After cooling, the mixture was con- centrated in vacuo to afford the title compound.
HPLC-MS (Method A): m/z: 298 (M+1 }; Rt = 4.54 min.
The compounds in the following examples were similarly prepared. Optionally, the com- pounds can be further purified by filtration and washing with water, ethanol and / or heptane instead of concentration in vacuo. Also optionally the compounds can be purified by washing with ethanol, water and/or heptane, or by chromatography, such as preparative HPLC. Example 67 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)thiazolidine-2,4-dione
Figure imgf000126_0002
HPLC-MS (Method C): m/z: 249 (M+1); Rt = 4.90 min
Example 68 (General procedure (B)) 5-Naphthalen-1-ylmethyleπethiazolidine-2,4-dione
Figure imgf000126_0003
HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4,16 min.
Example 69 (General procedure (B)) 5-Benzylidene-thiazolidine-2,4-dione 5 053070
126
Figure imgf000127_0001
HPLC-MS (Method A): m/z: 206 (M+1); Rt = 4,87 min.
Example 70 (General procedure (B)) 5-(4-Dtethylaminobenzy1idene)ihiazQlidine-2,4-clione
Figure imgf000127_0002
HPLC-MS (Method A): m/z: 277 (M+1); Rt = 4.73 min.
Example 71 (General procedure (B)) 5-(4-Methoxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000127_0003
HPLC-MS (Method A): m/r. 263 (M+1); Rt = 4,90 min.
Example 72 (General procedure (B)) 5-(4-Chloro-benzylidene)-thiazolidine-2,4*dione
Figure imgf000127_0004
HPLC-MS (Method A): m/z: 240 (M+1); Rt = 5,53 min.
Example 73 (General procedure (B)) 5-(4-Nitro-benzylidene)-thia2θlidine-2,4-dione
Figure imgf000128_0001
HPLC-MS (Method A): m/z: 251 (M+1); Rt = 4,87 min.
Example 74 (General procedure (B)) 5-(4-Hydroxy-3-methoxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000128_0002
HPLC-MS (Method A): m/z: 252 (M+1); Rt = 4,07 min.
Example 75 (General procedure (B)) 5-(4-Methylsulfanylbeπzylidene)thiazolidine-2,4-dione
Figure imgf000128_0003
HPLC-MS (Method A): m/z: 252 (M+1); Rt= 5,43 min.
Example 76 (General procedure (B)) 5-(2-Pentyloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000128_0004
HPLC-MS (Method C): m/z: 292 (M+1); Rt = 4.75 min.
1H NMR (DMSO-dβ): δ = 0.90 (3H, t), 1.39 (4H, m). 1.77 (2H, p). 4.08 (2H, t). 7.08 (1H, t),
7.14 (1H, d). 7.43 (2H1 m), 8.03 (1H. S). 12.6 (1H, bs).
Example 77 (General procedure (B)) 5-(3-Fluoro-4-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000129_0001
HPLC-MS (Method A): m/z: 354 (M+1); Rt = 4,97 miπ.
Example 78 (General procedure (B)) 5-(4-tert-Butylbenzylidene)thiazolidine-2,4-dioπe
Figure imgf000129_0002
HPLC-MS (Method A): m/z: 262 (M+1); Rt = 6,70 min.
Example 79 (General procedure (B)) Λ/-[4-(2,4-Dioxothiazolidiπ-5-ylidenemethy0pheπyI]acetanriide
Figure imgf000129_0003
HPLC-MS (Method A): m/z: 263 (M+1); Rt = 3,90 min.
Example 80 (General procedure (B)) 5-Biphenyl-4-ylmethylene-thiazolidiπe-2,4-dione
Figure imgf000129_0004
HPLC-MS (Method A): m/z: 282 (M+1); Rt = 4.52 min.
Example 81 (General procedure (B)) 5-(4-Phenoxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000129_0005
HPLC-MS (Method A): m/z: 298 (M+1); Rt = 6.50 min. Example 82 (General procedure (B)) 5-(3-Ben2yloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000130_0001
HPLC-MS (Method A): m/z: 312 (M+1); Rt = 6,37 min.
Example 83 (General procedure (B)) 5-(3-p-Tolyloxybenzylidene)thiazolidine-2,4-dione
Figure imgf000130_0002
HPLC-MS (Method A): m/z: 312 (M+1); Rt = 6,87 min.
Example 84 (General procedure (B)) 5-Naphthalen-2-ylmethylene-thiazolidine-2,4-dione
Figure imgf000130_0003
HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4.15 min.
Example 85 (General procedure (B)) 5-Benzo[1,3]dioxol-5-ylmethylenethiazolidine-2,4-dione
Figure imgf000130_0004
HPLC-MS (Method A): m/z: 250 (M+1), Rt = 3.18 min.
Example 86 (General procedure (B)) 5-(4-Chlorobenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000130_0005
HPLC-MS (Method A): m/z: 256 (M+1); Rt = 4,51 min.
Example 87 (General procedure (B)) 5-(4-Dimethylaminobenzylidene)-2-thioκothiazolidin-4-one
Figure imgf000131_0001
HPLC-MS (Method A): m/z: 265 (M+1); Rt = 5,66 min.
Example 88 (General procedure (B)) 5-(4-Nitrobeπzylidene)-2-thioxothiazolidin-4-one
Figure imgf000131_0002
HPLC-MS (Method A): m/z: 267 (M+1); Rt = 3,94 min.
Example 89 (General procedure (B)) 5-(4~Methylsulfany1benzylidene)-2-thtoxothiazolidin-4-one
Figure imgf000131_0003
HPLC-MS (Method A): m/z; 268 (M+1); Rt= 6,39 min.
Example 90 (General procedure (B)) 5-(3-Fluoro-4-methoxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000131_0004
HPLC-MS (Method A): m/z: 270 (M+1); Rt = 5,52 min.
Example 91 (General procedure (B)) 5-Naphthalen-2-ylmethylene-2-thioxσthiazo!idin-4-one
Figure imgf000132_0001
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 6,75 min.
Example 92 (Genera) procedure (B)) 5-(4-Diethylaminobenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000132_0002
HPLC-MS (Method A): m/z: 293 (M+1); Rt = 5,99 min.
Example 93 (General procedure (B)) 5-Biphenyl-4-ylmethylene-2-thioxothiazolidin-4-one
Figure imgf000132_0003
HPLC-MS (Method A): m/z: 298 (M+1); Rt= 7,03 min.
Example 94 (General procedure (B)) 5-(3-Phenoxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000132_0004
HPLC-MS (Method A): m/z: 314 (M+1); Rt = 6,89 min.
Example 95 (General procedure (B)) 5-(3-Benzyloxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000132_0005
HPLC-MS (Method A): m/z: 328 (M+1); Rt = 6,95 min. Example 96 (General procedure (B)) 5-(4-Benzyloxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000133_0001
HPLC-MS (Method A): m/z: 328 <M+1); RT = 6,89 min.
Example 97 (General procedure (B)) 5-Naphthalen-1-ylmethyleπe-2-thioxothiazolidin-4-one
Figure imgf000133_0002
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 6,43 min.
Example 98 (General procedure (B)) 5-(3-Methoxybenzyl)thiazolidine-2,4-dione
Figure imgf000133_0003
HPLC-MS (Method A): m/z: 236 (M+1); Rt = 3,05 min.
Example 99 (General procedure (D)) 4-[2-Chloro-4~(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid ethyl ester
O CH,
HPLC-MS (Method A): m/z: 392 (M+23), Rt = 4.32 min.
Example 100 (General procedure (D)) 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)-phenoxy]-butyric acid
Figure imgf000133_0004
HPLC-MS (Method A): m/z: 410 (M+23); Rt = 3,35 min. Example 101 (Genera! procedure (B)) 5-(3-Bromobenzylidene)thiazolidJne-2,4-dione
Figure imgf000134_0001
HPLC-MS (Method A): m/z: 285 (M+1 ); Rt = 4.01 min.
Example 102 (General procedure (B)) 5-(4-Bromobenzylidene)thiazolidine-2,4-dione
Figure imgf000134_0002
HPLC-MS (Method A): m/z: 285 (M+1 ); Rt - 4.05 min.
Example 103 (General procedure (B)) 5-(3-Ch[orobeπzylidene)thiazolidine-2,4-dioπe
Figure imgf000134_0003
HPLC-MS (Method A): m/z: 240 (M+1); Rt = 3.91 min.
Example 104 (General procedure (B)) 5-Thiophen-2-ylmethyleπethiazolidine-2,4-dione
Figure imgf000134_0004
HPLC-MS (Method A): m/z: 212 (M+1); Rt = 3.09 min.
Example 105 (General procedure (B)) 5-(4-BromothJophen-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000135_0001
HPLC-MS (Method A): m/z: 291 (M+1); Rt = 3.85 min.
Example 106 (General procedure (B)) 5-{3,5-DichlθrobenzyIidene)thiazolidine-2,4-dione
Figure imgf000135_0002
HPLC-MS (Method A): m/z: 274 (M+1); Rt = 4.52 min.
Example 107 (General procedure (B)) 5-(1 -Methyl-1 H-iπdol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000135_0003
HPLC-MS (Method A): m/z: 259 (M+1); Rt = 3.55 min.
Example 108 (Genera! procedure (B)) 5-(1H-(ndol-3-ylmethytene)thiazolidine-2,4-dione
Figure imgf000135_0004
HPLC-MS (Method A): m/z: 245 (M+1); Rt = 2.73 min.
Example 109 (General procedure (B)) 5-Fluoren-9-ylidenethiazolidine-2,4-dione
Figure imgf000136_0001
HPLC-MS (Method A): m/z: 280 (M+1); Rt = 4.34 min.
Example 110 (General procedure (B)) 5-(1-Phenylethylidene)thiazolidiπe-2,4-dione
Figure imgf000136_0002
HPLC-MS (Method A): m/z: 220 (M+1); Rt = 3,38 min.
Example 111 (General procedure (B)) 5-[1-(4-Methoxyphenyl)-ethylidene]-thia2olidine-2,4-dione
Figure imgf000136_0003
HPLC-MS (Method A): m/z: 250 (M+1); Rt = 3.55 min.
Example 112 (General procedure (B)) 5-(1 -Naphihalen-2-yl-ethylidene)-th/azolidine-2,4-dione
Figure imgf000136_0004
HPLC-MS (Method A): m/z: 270 (M+1); Rt = 4,30 min.
Example 113 (General procedure (B)) 5-[1-(4-Bromophenyl)-ethylidene]-thiazolidine-2,4-dioπe
Figure imgf000137_0001
HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,18 min.
Example 114 (General procedure (B)) 5-(2,2-Diphenylethylideπe)-thiazolidine-2,4-dione
Figure imgf000137_0002
HPLC-MS (Method A): m/z: 296 (M+1); Rt = 4,49 min.
Example 115 (General procedure (B)) 5-[1-(3-Methoxyphenyl)-ethylideπe]-thiazolidine-2,4-dione
Figure imgf000137_0003
HPLC-MS (Method A): m/z: 250 (M+1); Rt = 3,60 min.
Example 116 (General procedure (B)) 5-[1-(6-Methoxynaphthalen-2-yl)-ethy1idene]-thiazolidine-2,4-dione
Figure imgf000137_0004
HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,26 min.
Example 117 (General procedure (B)) 5-[1 -(4-Pheπoxyphenyl)-ethylidene]-lhiazolid ine-2,4-dione
Figure imgf000137_0005
HPLC-MS (Method A): m/z: 312 (M+1); Rt = 4,68 min.
Example 118 (General procedure (B)) 5-[1-(3-Fluoro-4-methoxyphenyl)ethylidene]thiazolidine-2,4-dione
Figure imgf000138_0001
HPLC-MS (Method A): m/z: 268 (M+1); Rt = 3,58 min.
Example 119 (General procedure (B)) 5-[1-(3-Brαmoρhenyl)-ethylidene]-thiazolidine-2,4-dione
Figure imgf000138_0002
HPLC-MS (Method A): m/z: 300 (M+1); Rt = 4,13 min.
Example 120 (General procedure (B)) 5-Anthracen-9-ylmethylenethiazαlidine-2,4-dione
Figure imgf000138_0003
HPLC-MS (Method A): m/z: 306 (M+1); Rt = 4,64 min.
Example 121 (General procedure (B)) 5-(2-Methoxynaphthalen-1-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000138_0004
HPLC-MS (Method A): m/z: 286 (M+1); Rt = 4,02 min. Example 122 (General procedure (B)) 5-(4-Methoxynaphthalen-1-ylmethyleπe)-thiazolidine-2,4-dione
Figure imgf000139_0001
HPLC-MS (Method A): m/z: 286 (M+1); Rt= 4,31 min.
Example 123 (General procedure (B)) 5-(4-Dimethylaminonaphthalen-1-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000139_0002
HPLC-MS (Method A): m/z: 299 (M+1); Rt = 4,22 min.
Example 124 (General procedure (B)) 5-(4-Methylnaphthalen-1-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000139_0003
HPLC-MS (Method A): m/z: 270 (M+1); Rt = 4,47 min.
Example 125 (General procedure (B)) 5-Pyridin-2-ylmethylene-thiazolidine-2,4-dione
Figure imgf000139_0004
Example 126
5-Pyrϊdin-2-yImethyl-thiazolidine-2,4-dione
Figure imgf000140_0001
5-Pyridin-2-y1methylene-thiazolidine-2,4-dione (5 g) in tetrahydrofuran (300 ml) was added 10% Pd/C (1 g) and the mixture was hydrogenated at ambient pressure for 16 hours. More 10% Pd/C (5 g) was added and the mixture was hydrogenated at 50 psi for 16 hours. After filtration and evaporation in vacuo, the residue was purified by column chromatography etuting with a mixture of ethyl acetate and heptane (1 :1). This afforded the title compound (0.8 g, 16%) as a solid.
TLC: Rf = 0.30 (SiO2; EtOAc: heptane 1:1)
Example 127 (General procedure (B)) 5-(1H-lmidazol-4-yfmethyfene)-thiazolidine-2,4-dione
Figure imgf000140_0002
Example 128 (General procedure (B))
5-(4-Benzyloxy-benzylidene)-thiazolidine-2,4-dione
Figure imgf000140_0003
HPLC-MS (Method A): m/z: 6,43 min ; 99 % (2A)
Example 129 (General procedure (B))
5-[4-(4-Fluorobenzyloxy)benzylidene]-2-thioxothiazolidin-4-one
Figure imgf000140_0004
Example 130 (General procedure (B)) 5-(4-Butoxybenzylidene)-2-thioxothiazolidin-4-one
Figure imgf000141_0001
Example 131 (General procedure (B)) 5-(3-Methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000141_0002
HPLC-MS (Method A): m/z: 236 (M+1); Rt = 4,97 min
Example 132 (General procedure (B)) 5-(3-Methoxybenzylidene)imidazolidine-2.4-dione
Figure imgf000141_0003
HPLC-MS (Method A): m/z: 219 (M+1); Rt = 2.43 min.
Example 133 (General procedure (B)) 5-(4-Methoxybenzylidene)imidazolidine-2,4-dione
Figure imgf000141_0004
HPLC-MS (Method A): m/z: 219 (M+1); Rt = 2.38 min. Example 134 (General procedure (B)) 5-(2,3-Dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000142_0001
Example 135 (General procedure (B)) 5-Benzofuran-7-ylrnethylenethiazolidine-2,4-dione
Figure imgf000142_0002
HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4,57 min.
Example 136 (General procedure (B)) 5-Benzo[1,3]dioxol-4-ylmethylenethiazolidine-2,4-dione
Figure imgf000142_0003
HPLC-MS (Method C): m/z: 250 (M+1); Rt = 4,00 min.
Example 137 (General procedure (B)) 5-(Φ-Methoxy-2,3-dimethylbenzylidene)thiazolidine-2,4-dione
Figure imgf000142_0004
HPLC-MS (Method C): m/z: 264 (M+1); Rt = 5,05 min.
Example 138 (General procedure (B)) 5-(2-Benzyloxy-3-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000142_0005
HPLC-MS (Method C): m/z: 342 (M+1); Rt = 5,14 min. Example 139 (General procedure (B)) 5-(2-Hydroxybenzylidene)thiazolidine-2,4-dione
Figure imgf000143_0001
HPLC-MS (Method C): m/z: 222 (M+1); Rt = 3,67 min.
Example 140 (General procedure (B)) 5-(2,4-Dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000143_0002
1H-NMR (DMSO-cfe): 7.60 (2H, "s"), 7.78 (1H. s), 7.82 (1H, s).
Example 141 (General procedure (B)) 5-(2-Chlorobenzylideπe)thiazσlidine-2,4-dione
Figure imgf000143_0003
1H-NMR (DMSO-d6): 7.40 (1H, t), 7.46 (1H, t), 7.57 (1H, d), 7.62 (1H, d), 7.74 (1H, s).
Example 142 (General procedure (B)) 5-(2-Bromobenzylideπe)thiazolidiπe-2,4-dioπe
Figure imgf000143_0004
1H-NMR (DMSOd6): 7.33 (1H, t), 7.52 (1H. t), 7.60 (1H. d). 7.71 (1H, s), 7.77 (1H, d).
Example 143 (General procedure (B)) 5-(2,4-Dimethoxybenzy!idene)thiazolidine-2,4-dione
Figure imgf000143_0005
HPLC-MS (Method C): m/z: 266 (M+1) Rt = 4.40 min. Example 144 (General procedure (B)) 5-(2-Methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000144_0001
HPLC-MS (Method C): m/z: 236 (M+1 ); Rt = 4,17 min.
Example 145 (General procedure (B)) 5-(2,6-Difluorobenzylidene)thiazolidine-2,4-dione
Figure imgf000144_0002
HPLC-MS (Method C): m/z: 242 (M+1 ); Rt = 4.30 min.
Example 146 (General procedure (B)) 5-(2,4-Dimethylbenzylidene)thiazolidine-2,4-dioπe
Figure imgf000144_0003
HPLC-MS (Method C): m/z: 234 (M+1); Rt = 5,00 min.
Example 147 (General procedure (B)) 5-(2,4,6-Trimethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000144_0004
HPLC-MS (Method C): m/z: 296 (M+1 ); Rt = 4,27 min.
Example 148 (General procedure (B)) 5-(4-Hydroxy-2-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000144_0005
HPLC-MS (Method C): m/z: 252 (M+1); Rt = 3,64 min.
Example 149 (General procedure (B)) 5-(4-Hydroxynaphthaleπ-1-ylmethylene)thiazolidine-2,4-dione
Figure imgf000145_0001
1H-NMR (DMSO-cfe): 6 = 7.04 (1H1 d), 7.57 (2H. m), 7.67 (1H. t), 8.11 (1H, d), 8.25 (1H, d), 8.39 (1H, s) 11.1 (1H, s), 12.5 (1H. bs). HPLC-MS (Method C): m/z: 272 (M+1); Rt = 3.44 min.
Example 150 (General procedure (B))
5-(2-Trifluorornethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000145_0002
HPLC-MS (Method C): m/z: 290 (M+1); Rt = 4,94 min.
Example 151 (General procedure (B))
5-Biphenyl-2-ylmethylenethiazolidine-2,4-dione
Figure imgf000145_0003
HPLC-MS (Method C): m/z: 282 (M+1); Rt = 5,17 min.
Example 152 (General procedure (B))
5-(2-Benzyloxybeπzylidene)thiazolidiπe-2,4-dione
Figure imgf000145_0004
HPLC-MS (Method C): m/z: 312 (M+1); Rt = 5,40 min.
Example 153 (General procedure (B)) 5-Adamantan-2-ylidenethiazolidine-2,4-dione
Figure imgf000146_0001
HPLC-MS (Method A): m/z: 250 (M+1); Rt = 4,30 min.
Example 154 {General Procedure (B))
5-[3-(4-Nitrophenyl)allyIidene]thiazolidiπe-2,4-dioπe
Figure imgf000146_0002
HPLC-MS (Method C): m/z: 277 (M+1); Rt = 3.63 min.
Example 155 (General Procedure (B)) 5-[3-(2-Methoxyphenyl)allyIidenelthiazolidiπe-2,4-dione
Figure imgf000146_0003
HPLC-MS (Method C): m/z: 262 (M+1 ); Rt = 3.81 min.
Example 156 (General Procedure (B)) 5-[3-(4-Methoxyphenyl)allylidene]thiazolidine-2.4-dione
Figure imgf000146_0004
HPLC-MS (Method C): m/z: 262 (M+1); Rt= 3.67 min. Example 157 (General procedure (B)) 5-(4-Hydroxybenzylidene)thiazolicline-214-dione
Figure imgf000147_0001
Example 158 (General procedure (B))
5-(4-Dimethylaminobenzylidene)pyrimidine-2,4,6-trione
Figure imgf000147_0002
HPLC-MS (Method C): m/z = 260 (M+1) Rt = 2.16 min.
Example 159 (General procedure (B))
5-(9-Ethyl-9H-carbazol-2-ylmethylene)-pyrimidine-2,4,6-trione
Figure imgf000147_0003
HPLC-MS (Method C): m/z = 334 (M+1); Rt = 3,55 min.
Example 160 (General procedure (B))
5-(4-Hexy]oxynaphthalen-1-ylmethylene)thiazolidiπe-2,4-dione
Figure imgf000147_0004
HPLC-MS (Method C): m/z = 356 (M+1 ); Rt = 5.75 min. Example 161 (General procedure (B)) 5-(4-Decyloxynaphthalen-1-ylmethylene)thiazolidiπe-2,4-dione
Figure imgf000148_0001
HPLC-MS {Method C): m/z = 412 (M+1); Rt = 6.44 min.
Example 162 (General procedure (B)) 5-[4-(2-Aminoethoxy)-naphthalen-1-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000148_0002
HPLC-MS (Method C): m/z = 315 (M+1); Rt = 3,24 min.
Example 163 (General procedure (B)) 5-(2,4-Dimethyl-9H-carbazol-3-ylmethylene)-pyrimidine-214,6-trione
Figure imgf000148_0003
HPLC-MS (Method C): m/z = 334 (M+1); Rt = 3,14 min.
Example 164 (General procedure (B)) 4-(4-Hydroxy-3-methoxybenzylidine)hydantoin
Figure imgf000148_0004
Example 165 (General procedure (B)) 5-Benzylidenehydantoin
Figure imgf000149_0001
General procedure (C) for preparation of compounds of general formula I2:
Figure imgf000149_0002
I2 wherein X1 Y, A, and R3 are as defined above and A is optionally substituted with up to four substituents R7, R8, Rβ, and R10 as defined above.
This general procedure (C) is quite similar to general procedure (B) and is further illustrated in the following example:
EΞxample 166 (General procedure (C)) 5-(3,4-Dibromobenzylidene)thiazolidine-2,4-dione
Figure imgf000149_0003
A mixture of thiazolidine-2,4-dione (90%, 65 mg, 0.5 mmol), 3,4-dibromobenzaldehyde (132 mg, 0.5 mmol), and piperidiπe (247 μL, 2.5 mmol) was shaken in acetic acid (2 ml_) at 1100C for 16 hours. After cooling, the mixture was concentrated to dryness in vacuo . The resulting crude product was shaken with water, centrifuged, and the supernatant was discarded. Subsequently the residue was shaken with ethanol, centrifuged, the supernatant was discarded and the residue was further evaporated to dryness to afford the title com¬ pound. 1H NMR (Acetoπe-dβ): <*H 7.99 (d,1H). 7.90 (d,1H). 7.70 (s,1H), 7.54 (d,1H); HPLC-MS (Method A): m/z: 364 (M+1); Rt = 4.31 min.
The compounds in the following examples were similarly prepared. Optionally, the com- pounds can be further purified by filtration and washing with water instead of concentration in vacuo. Also optionally the compounds can be purified by washing with ethanol, water and/or heptane, or by preparative HPLC.
Example 167 (General procedure (C)) 5-(4-Hydroxy-3-iodo-5-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000150_0001
Mp = 256 0C; 1H NMR (DMSO-dβ) δ ~ 12.5 (s,broad,1H), 10.5 (s.broad.iH), 7.69 (s,1H), 7.51 (d,1H), 7.19 (d,1H)3.88 (s,3H). 13C NMR (DMSO-Cf6) δc = 168.0, 167.7 . 149.0, 147.4, 133.0, 131.2, 126.7, 121.2, 113.5, 85.5, 56.5; HPLC-MS (Method A): m/z: 378 (M+1); Rt = 3.21 min.
Example 168 (General procedure (C)) 5-(4-Hydroxy-2,6-dimethylbenzylidene)thiazolidine-2,4-dione
Figure imgf000150_0002
HPLC-MS (Method C): m/z: 250 (M+1 ); Rt.= 2.45 min.
Example 169 (General procedure (C)) 4-[5-Bromo-6-(2,4-dioxothiazolidiπ-5-ylideπemethyl)-naphthalen-2-yloxymethyl]-benzoic acid
Figure imgf000150_0003
HPLC-MS (Method C): m/z: 506 (M+23); Rt.= 4.27 min. Example 170 (General procedure (C)) 5-(4-Bromo-2,6-dichlorαbenzylidene)thiazolidine-2,4-dione
Figure imgf000151_0001
HPLC-MS (Method C): m/z: 354 (M+1); Rt.= 4.36 min.
Example 171 (General procedure (C)) 5-(6-Hydroxy-2-naphthylmethyleπe) thiazolidiπe-2,4-dione
Figure imgf000151_0002
Mp 310-314 0C, 1H NMR (DMSO-cfe): δH = 12.5 (s,broadt1H), 8.06(d,1H), 7.90-
7.78(m,2H),7.86 (s,1H), 7.58 (dd,1H),7.207.12 (m,2H). 13C NMR (DMSOd6): δc = 166.2, 165.8 , 155.4, 133.3, 130.1, 129.1, 128.6, 125.4, 125.3, 125.1, 124.3, 120.0, 117.8, 106.8; HPLC-MS (Method A): m/z: 272 (M+1); Rt = 3.12 min.
Preparation of the starting material, 6-hydroxy-2-naphtalenecarbaldehyde:
6-Cyano-2-naphthaleπecarbaldehyde (1.0 g. 5.9 mmol) was dissolved in dry hexane (15 mL) under nitrogen. The solution was cooled to -600C and a solution of diisobutyl aluminium hy- dride (DIBAH) (15 mL, 1M in hexane) was added dropwise. After the addition, the solution was left at room temperature overnight. Saturated ammonium chloride solution (20 mL) was added and the mixture was stirred at room temperature for 20 min, subsequently aqueous H2SO4 (10% solution. 15 mL) was added followed by water until all salt was dissolved. The resulting solution was extracted with ethyl acetate (3x), the combined organic phases were dried with MgSO4, evaporated to dryness to afford 0.89 g of 6-hydroxy-2- naphtalenecarbaldehyde.
Mp.: 153.5-156.5 °°; HPLC-MS (Method A): m/z: 173 (M+1); Rt = 2.67 min; Λ H NMR (DMSO- dβ): SH = 10.32(s,1H), 8.95 (d,1H)r 10.02 (s,1H), 8.42 (s,broad,1H). 8.01 (d,1H), 7.82-7.78 (m,2H), 7.23-7.18 (m,2H). Alternative preparation of 6-hydroxy-2-naphtalenecarbaldehyde: To a stirred cooled mixture of 6-bromo-2-hydroxynaphthalene (25.3 g, 0.113 mol) in THF (600 mL) at -780C was added n-BuLi (2.5 M, 100 mLr 0.250 mol) dropwise. The mixture turned yellow and the temperature rose to -640C. After ca 5 miπ a suspension appeared. After addition, the mixture was maintained at -780C. After 20 minutes, a solution of DMF (28.9 mL, 0.373 mol) in THF (100 mL) was added over 20 minutes. After addition, the mix¬ ture was allowed to warm slowly to room temperature. After 1 hour, the mixture was poured in ice/water (200 mL). To the mixture citric acid was added to a pH of 5. The mixture was stirred for 0.5 hour. EthyJ acetate (200 mL) was added and the organic layer was separated and washed with brine (100 mL), dried over Na2S04and concentrated. To the residue was added heptane with 20% ethyl acetate (ca 50 mL) and the mixture was stirred for 1 hour. The mixture was filtered and the solid was washed with ethyl acetate and dried in vacuo to afford 16 g of the title compound.
Example 172 (General procedure (C))
5-(3-lodo-4-methoxybenzylidene)thiazolidieπe-2,4-dione
Figure imgf000152_0001
1H NMR (DMSO-dβ): δH 12.55 (s,broad,1H), 8.02 (d,1H), 7.72 (s,1H). 7.61 (d,1H)7.18(d,iH). " 3.88 (s,3H); 13C NMR (DMSO-C-8): δc 168.1, 167.7 , 159.8, 141.5" 132.0. 130.8, 128.0, 122.1, 112.5, 87.5, 57.3. HPLC-MS (Method A): m/z: 362 (M+1 ); Rt = 4.08 min.
Preparation of the starting material, 3-iodo-4-methoxybenzaldehyde:
4-Methoxybenzaldehyde (0.5 g, 3.67 mmol) and silver trifluoroacetate (0.92 g, 4.19 mmol) were mixed in dichloromethane (25 mL). Iodine (1.19 g, 4.7 mmol) was added in small por¬ tions and the mixture was stirred overnight at room temperature under nitrogen. The mixture was subsequently filtered and the residue washed with DCM. The combined filtrates were treated with an acqueous sodium thiosulfate solution (1 M) until the colour disappeared. Subsequent extraction with dichloromethane (3 x 20 mL) followed by drying with MgSO4 and evaporation in vacuo afforded 0.94 g of 3-iodo-4-methoxybeπzaldehydβ.
Mp 104-107 0C; HPLC-MS (Method A): m/z:263 (M+1); Rt = 3.56 min.;*H NMR (CDCl3): δH = 8.80 (s,1H), 8.31 (d,1H), 7.85 (dd.1H) 6.92 (d,1H). 3.99 (s, 3H). Example 173 (General procedure (C))
5-( 1 -Bromonaphthalen-2-ylmethylene)thiazolidine-2,4-dione
Figure imgf000153_0001
HPLC-MS (Method A): m/z: =336 (M+1); Rt = 4.46 min.
Example 174 (General procedure (C)) 1-I5-(2,4-Dioxothiazolidin-5-ylidenemethyl)thiazol-2-yl]piperidine-4-carboxy|jc acid ethyl ester
Figure imgf000153_0002
1H NMR (DMSO-cfe): δH = 7.88 (s.1H), 7.78 (s,1H).4.10 (q,2H), 4.0-3.8 (m.2H), 3.40-3.18 (m,2H), 2.75-2.60 (m,iH). 2.04-1.88 (m,2H), 1.73-1.49 (m,2H) , 1.08 (t,3H); HPLC-MS (Method A): m/z: 368 (M+1); Rt * 3.41 min.
Example 175 (General procedure (C))
5-(2-Phenyl-[1 ,2,3]triazol-4-ylmethylene) thiazolidine-2,4-dione
Figure imgf000153_0003
1H NMR (DMSO-cfe): δH = 12.6 (s,broad,1H), 8.46 (s,1H), 8.08 (dd,2H), 7.82 (s,1H). 7.70-7.45 (m, 3H). HPLC-MS (Method A): m/z: 273 (M+1); Rt = 3.76 min.
Example 176 (General procedure (C)) 5-(Quinolin-4-ylrnethylene)thiazolidine-2,4-dione
Figure imgf000153_0004
HPLC-MS (Method A): m/z: 257 (M+1); Rt = 2.40 min. Example 177 (General procedure (C)) 5-(6-Methylpyridin-2-ylrnethylene)triiazolidine-2,4-dione
Figure imgf000154_0001
1H NMR (DMSO-dβ): *H = 12.35 (s.broad.iH), 7.82 (t,1H), 7.78 (S,1H), 7.65 (d,1H).7.18 (d,1H), 2.52 (s,3 H); HPLC-MS (Method A): m/z: 221 (M+1); Rt - 3.03 min.
Example 178 (General procedure (C)) 5-(214-dioxothJa2olidjn-5-yfidenemethyl)-furan-2-ylmethyla∞tate
Figure imgf000154_0002
1H NMR (DMSO-dβ): δH = 12.46 (s,broad,1H), 7.58 (s,1H), 7.05 (d,1H), 6.74 (s,1H), 5.13 (s,2H), 2.10 (s,3H). HPLC-MS (Method A): m/z: 208 (M-CH3COO); Rt = 2.67 min.
Example 179 (General procedure (C)) 5-(2,4-Dioxothia2olidin-5-ylidenemethy!)fυran-2-sulfonic acid
Figure imgf000154_0003
HPLC-MS (Method A): m/z:276 (M+1); Rt = 0.98 min.
Example 180 (General procedure (C)) 5-(5-Benzyloxy-1H-pyrrolo[2,3-clpyridin-3-ylmethylene)-thiazorιdine-2,4-dione
Figure imgf000154_0004
HPLC-MS (Method A): m/z: 352 (M+1); Rt = 3.01 min.
Example 181 (General procedure (C)) 5-(Quinoliπ-2-ylmethylene)thiazolidine-2.4-dione
Figure imgf000155_0001
HPLC-MS (Method A): m/z: 257 (M+1); Rt = 3.40 miπ.
Example 182 (General procedure (C)) 5-(2t4-Dioxothia2olidin-5-ylideπemethyl)thiopheπe-2-carboxylic acid
Figure imgf000155_0002
HPLC-MS (Method A): m/z: 256 (M+1); Rt = 1.96 min.
Example 183 (General procedure (C)) 5-(2-Phenyl-1 H-imidazol-4-ylmethylene)thiazolidine-2,4-dione
Figure imgf000155_0003
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 2.89 min.
Example 184 (General procedure (C)) 5-(4-lmidazo[-1-yl-benzy[idene)thiazolidine-2,4-dione
Figure imgf000155_0004
HPLC-MS (Method A): m/z: 272 (M+1); Rt = 1.38 min.
Example 185 (General procedure (C)) 5-(9-Ethyl-9H-carbazol-3-ylmethy]eπe)thiazolidine-2,4-dione
Figure imgf000155_0005
HPLC-MS (Method A): m/z: 323 (M+1); Rt = 4.52 min. Example 186 (General procedure (C)) 5-(1,4-Dimethyl-9H-carbazol-3-ylrnethylene)thiazolidine-2,4-dione
Figure imgf000156_0001
HPLC-MS (Method A): m/z: 323 (M+1); Rt = 4.35 min.
Example 187 (General procedure (C)) 5-(2-Methyl-1H-indol-3-y1methylene)thiazolidine-2,4-dione
Figure imgf000156_0002
HPLC-MS (Method A): m/z: 259 (M+1 ); Rt = 3.24 min.
Example 188 (General procedure (C)) 5-(2-Ethylindol-3-ylmethylene)thiazolidtne-2,4-dione
Figure imgf000156_0003
2-Methyltndote (1.0 g, 7.6mmol) dissolved in diethyl ether (100 mL) under nitrogen was treated with n-Butyl lithium (2 M in pentane, 22.8 mmol) and potassium ferf-butoxide (15.2 mmol) with stirring at RT for 30 min. The temperature was lowered to -70 C and methyl Io¬ dide (15.2 mmol) was added and the resulting mixture was stirred at -70 for 2 h. Then 5 drops of water was added and the mixture allowed to warm up to RT. Subsequently, the mix- ture was poured into water (300 mL), pH was adjusted to 6 by means of 1N hydrochloric acid and the mixture was extracted with diethyl ether. The organic phase was dried with Na2SO4 and evaporated to dryness. The residue was purified by column chromatography on silica gel using heptane/ether( 4/1) as eluent. This afforded 720 mg (69 %) of 2-ethylindole.
1H NMR (DMSO-rfβ): δ = 10.85 (1H,S); 7.39 (1H,d); 7.25 (1H,d); 6.98(1 H,t); 6.90(1 H,t); 6.10 (1H,s); 2.71 (2H,q); 1.28 (3H,t). 2-Ethylindole (0.5 g, 3.4mmol) dissolved in DMF (2 mL) was added to a cold (0 0C) premixed (30 minutes) mixture of DMF (1.15 mL) and phosphorous oxychloride (0.64 g, 4.16 mmol). After addition of 2-ethylindole, the mixture was heated to 400C for 1 h, water (5 mL) was added and the pH adjusted to 5 by means of 1 N sodium hydroxide.The mixture was subse- quently extracted with diethyl ether, the organic phase isolated, dried with MgSO4 and evapo¬ rated to dryness affording 2-ethylindole-3-carbaldehyde (300 mg ).
HPLC-MS (Method C): m/z:174 (M+1); Rt. =2.47 min.
2-EthylindoIe-3-carbaldehyde (170 mg) was treated with thiazolidine-2,4-dioπe using the general procedure (C) to afford the title compound (50 mg).
HPLC-MS (Method C):m/z: 273 (M+1); Rt - 3.26 min.
Example 189 (General procedure (C))
5-[2-(4-Bromophenylsulfanyl)-1-methyl-1H-indol-3-ylmethylene]thiazolidine-2,4-dione
Figure imgf000157_0001
HPLC-MS (Method A): m/z: 447 (M+1); Rt - 5.25 min.
Example 190 (General procedure (C))
5-[2-(2,4-Dichlorobenzyloxy)-naphthalen-1-ylmethylene]thiazolidine-2,4-dione
Figure imgf000157_0002
HPLC-MS (Method A): (anyone 1) m/z: 430 (M+1); Rt= 5.47 min. Example 191 (General procedure (C)) 5-{4-[3-(4-Bromophenyl)-3-oxopropenyl]-benzylidene}thia2olidine-2,4-dioπe
Figure imgf000158_0001
HPLC-MS (Method A): m/z: 416 (M+1); Rt = 5.02 min.
Example 192 (General procedure (C)) 5-(4-Pyridin-2-ylbeπzylidene)thiazolidiπe-2,4-dione
Figure imgf000158_0002
HPLC-MS (Method A): m/z: 283 (M+1). Rt = 2.97 min.
Example 193 (General procedure (C)) 5-(3,4-Bisbenzyloxybenzylidene)thiazo!idine-2,4-dione
Figure imgf000158_0003
HPLC-MS (Method A): m/z: 418 (M+1); Rt = 5.13 min.
Example 194 (General procedure (C)) 5-[4-(4-Nitrobenzyloxy)-benzylidene]thiazolidine-2,4-dioπe
Figure imgf000158_0004
HPLC-MS (Method A): m/z: 357 (M+1); Rt = 4.45 min.
Example 195 (General procedure (C)) 5-(2-Phenyl-1H-indol-3-ylmethyleπe)thiazolidine-2,4-dione
Figure imgf000159_0001
HPLC-MS (Method A): m/z: 321 (M+1); Rt = 3.93 min.
Example 196 (General procedure (C)) 5-(5-Benzyloxy-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000159_0002
HPLC-MS (Method A): m/z: 351 (M+1); Rt = 4.18 min.
Example 197 (General procedure (C)) 5-(4-Hydroxybenzyliderte)thiazo1idine-2,4-dione
Figure imgf000159_0003
HPLC-MS (Method A): m/z: 222 (M+1); Rt= 2.42 min. Example 198 (General procedure (C)) 5-(1-Methyl-1H-indol-2-ylmethylene)thiazolidine-2.4-dione
Figure imgf000159_0004
'H NMR (DMSO-d6): δH = 12.60 (s,broad,1H), 7.85 (s.1H), 7.68 (dd,1H), 7.55 (dd,1 H), 7.38 (dt,1H), 7.11 (dt,iH) 6.84 (s,iH), 3.88 (s,3H); HPLC-MS (Method A): m/z: 259 (M+1); Rt = 4.00 min.
Example 199 (General procedure (C))
5-(5-Nitro-1H-indo!-3-ylmethy[ene)thiazolidine-2.4-dione
Figure imgf000160_0001
.Mp 330-3330C, 1H NMR (DMSO-dβ): <5H = 12.62 (s,broad,1H), 8,95 (d,1H), 8.20 (s,1H), 8.12 (dd,1H), 7.98 (s.broad.iH), 7.68 (d,iH); HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.18 min.
Example 200 (Genera! procedure (C)) 5-(6-Methoxynaphthaten-2-ylmethy!ene)thiazolidine-2.4-dione
Figure imgf000160_0002
HPLC-MS (Method A): m/z: 286 (M+1); Rt = 4.27 min.
Example 201 (General procedure (C)) 5-(3-Bromo-4-methoxybenzylJdene)thiazolidine-2,4-dione
Figure imgf000160_0003
HPLC-MS (Method A): m/z: 314 (M+1), Rt = 3.96 min.
Example 202 (General procedure (C)) 3-{(2-Cyanoethyl)-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenyl]amino}propionitrile
Figure imgf000160_0004
HPLC-MS (Method A): mfe 327 (M+1); Rt = 2.90 min.
Example 203 (General procedure (C)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid methyl ester
Figure imgf000161_0001
HPLC-MS (Method A): m/z: 303 (M+1); Rt = 3.22-3-90 min.
Example 204 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-6-carboxylic acid pentyl ester.
Figure imgf000161_0002
3-(2,4-Dioxothiazolidin-5-y!idenemethyl)indole-6-carboxylic acid methyl ester (example 203, 59 mg; 0.195mmol) was stirred in pentanol (20 mL) at 1450C for 16 hours. The mixture was evaporated to dryness affording the title compound (69 mg).
HPLC-MS (Method C): m/z: 359 (M+1); Rt= 4.25 min.
Example 205 (General procedure (C)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)iπdole-7-carboxylic acid
Figure imgf000161_0003
HPLC-MS (Method A): m/z: 289 (M+1); Rt - 2.67 min.
Example 206 (General procedure (C)) 5-(1-Benzylindol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000161_0004
HPLC-MS (Method A): m/z: 335 (M+1): Rt = 4.55 min. Example 207 (General procedure (C)) 5-(1-Benzenesulfonylindot-3-yImethylene)thiazotidine-2,4-dione
Figure imgf000162_0001
HPLC-MS (Method A): m/z: = 385 (M+1); Rt = 4.59 min.
Example 208 (General procedure (C)) 5-(4-[1,2,3]Thiadiazol-4-ylben2ylidene)thiazo!idiπe-2,4-dione
Figure imgf000162_0002
HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.45 min.
Example 209 (General procedure (C)) 5-[4-(4-Nitrobenzyloxy)-benzylidene]thiazolidine-2,4-dione
Figure imgf000162_0003
HPLC-MS (Method A): m/z: 357 (M+1); Rt = 4.42 min.
Example 210 (General procedure (C)) 3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indole-1-carboxylic acid ethyl ester
Figure imgf000162_0004
HPLC-MS (Method A): m/z: 317 (M+1); Rt = 4.35 min. Example 211 (General procedure (C)) 5-[2'(4-Pentylbenzoy|)-beπzofuran-5-ylmethyIene]thiazolidiπe-2,4-dione
Figure imgf000163_0001
HPLC-MS (Method A): m/z: 420 (M+1); Rt = 5.92 miπ.
Example 212 (General procedure (C)) 5-[1-(2-Fluorobenzyl)-4-nitroindoI-3-ylmethylene]triiazolidine-2,4-dioπe
Figure imgf000163_0002
HPLC-MS (Method A): (Anyone 1) m/z: 398 (M+1); Rt = 4.42 min.
Example 213 (General procedure (C)) 5-(4-Benzyloxyindol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000163_0003
HPLC-MS (Method A): m/z: 351 (M+1); Rt = 3.95 min.
Example 214 (General procedure (C)) 5-(4-lsobutyIbenzylideneHniazolidine-2,4-dione
Figure imgf000163_0004
HPLC-MS (Method A): m/z: 262 (M+1); Rt = 4.97 min,
Example 215 (General procedure (C))
Trifluoromethanesurfoπrc acid 4-(2,4-droxothiazoltdrn-5-ylidenemethyl)naphthaleπ-1-yl ester
Figure imgf000164_0001
HPLC-MS (Method A): m/z: 404 (M+1 ); Rt = 4.96 min.
Preparation of starling material: 4-Hydroxy-1-naphthaldehyde (10 g, 58 mmol) was dissolved in pyridin (50 ml) and the mix¬ ture was cooled to 0-5 DC. With stirring, trifluoromethanesulfonic acid anhydride (11.7 ml, 70 mmol) was added drop-wise. After addition was complete, the mixture was allowed to warm up to room temperature, and diethyl ether (200 ml) was added. The mixture was washed with water (2 x 250 ml), hydrochloric acid (3N, 200 ml), and saturated aqueous sodium chloride (100 ml). After drying (MgSO4), filtration and concentration in vacuo, the residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4). This afforded 8.35 g (47%) trifluoromethanesulfonic acid 4-formylnaphtha!en-1 -yl ester, mp 44-46.6 0C.
Example 216 (General procedure (C))
5-(4-Nitroindol-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000164_0002
HPLC-MS (Method A): m/z: 290 (M+1); Rt = 3.14 min.
Example 217 (General procedure (C))
5-(3,5-Dibromo-4-hydroxy-benzylideπe)thiazo/idine-2,4-dione
Figure imgf000164_0003
1H NMR (DMSO-cfe): δH = 12.65 (broad,1H), 10.85 (broad,1H), 7.78 (S.2H), 7.70 (s,1H); HPLC-MS (Method A): m/z: 380 (M+1); Rt = 3.56 min. Example 218 (General procedure (C))
Figure imgf000165_0001
HPLC-MS (Method A): m/z: 385 (M+1); Rt = 5.08 min.
General procedure for preparation of starting materials for examples 218 - 221 : lndole-3-carbaldehyde (3.8 g, 26 mmol) was stirred with potassium hydroxide (1.7 g) in ace¬ tone (200 ml_) at RT until a solution was obtained indicating full conversion to the indole po¬ tassium salt. Subsequently the solution was evaporated to dryness in vacuo. The residue was dissolved in acetone to give a solution containing 2.6 mmol/20 mL.
20 mL portions of this solution were mixed with equimotar amounts of arylmethyibromides in acetone (10 mL). The mixtures were stirred at RT for 4 days and subsequently evaporated to dryness and checked by HPLC-MS. The crude products, 1-benzylated indole-3- carbaldehydes, were used for the reaction with thiazolidine-2,4-dione using the general pro¬ cedure C.
Example 219 (General procedure (C)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-ylmethyl]benzoic acid methyl ester
Figure imgf000165_0002
HPLC-MS (Method A): m/z: 393 (M+1); Rt = 4.60 min.
Example 220 (General procedure (C))
S-fi-tθ.iO-Dioxo-Θ.IO-dihydroanthracen^-ylmethylJ-IH-indol-S-ylmethyienelthiazolidine^Λ- dione
Figure imgf000165_0003
HPLC-MS (Method A): m/z: 465 (M+1); Rt = 5.02 min.
Example 221 (General procedure (C)) 4-[3-(2,4-Dioxothiazoltdin-5-ylidenemethyl)indol-1-ylmethyl]biphenyl-2-carbonitrile
Figure imgf000166_0001
HPLC-MS (Method A): m/z: 458 (M+23); Rt = 4.81 min.
Example 222 (General procedure (C)) 3-[3-(2,4-Dioxothiazo(idin-5-yIideπemethyl)-2-methylindol-1-ylmethyl]benzonitrile.
Figure imgf000166_0002
2-Methylindole-3-carbaldehyde (200 mg, 1.26 mmol) was added to a slurry of 3- bromomethylbenzenecarbonitrile (1.26 mmol) followed by sodium hydride, 60%, (1.26 mmol) in DMF (2 mL). The mixture was shaken for 16 hours, evaporated to dryness and washed with water and ethanol. The residue was treated with thiazolidine-2,4-dione following the general procedure C to afford the title compound (100 mg).
HPLC-MS (Method C): m/z: 374 (M+1); Rt. = 3.95 min.
Example 223 (General procedure (C)) 5-( 1 -Benzyl-2-methylindol-3-ylmethytene)thiazolidine-2,4-dione.
Figure imgf000166_0003
This compound was prepared in analogy with the compound described in example 222 from benzyl bromide and 2-methylindole-3-carbaldehyde, followed by reaction with thiazolidine- 2,4-dione resulting in 50 mg of the title compound. HPLC-MS (Method C): m/z: 349 (M+1); Rt. = 4.19 miπ.
Example 224 4-[3-(2,4-Dioxothiazolidin-5-ylic!enemethyl)-2-methylindol-1-ylmethyl]benzoic acid methyl es¬ ter
Figure imgf000167_0001
This compound was prepared in analogy with the compound described in example 222 from 4-(bτomomethyl)benzoic acid methyl ester and 2-methylindole-3-carbaldehyde, followed by reaction with thiazolidine-2,4-dione.
HPLC-MS (Method C): m/z: 407 (M+1); Rt.= 4.19 min.
Example 225 (General procedure (C)) m 5-(2-Chloro-1-methyl-1H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000167_0002
HPLC-MS (Method A): m/z: 293 (M+1); Rt = 4.10 min.
Example 226 (General procedure (C)) 5-(4-Hydroxy-3,5-diiodo-benzylidene)-thiazolidine-2,4-dione
Figure imgf000167_0003
HPLC-MS (Method A): m/z: 474 (M+1); Rt = 6.61 miπ.
Example 227 (General procedure (C)) 5-(4-Hydroxy-3-iodobenzylidene)thiazolidine-2,4-dione
Figure imgf000168_0001
HPLC-MS (Method C): m/z: 348 (M+1); Rt. = 3.13 min
1H-NMR: (DMSOd6 ): 11.5 (1H,broad); 7.95(1H,d); 7.65(1H,s); 7.45 (1H.dd); 7.01(1H,dd);
3.4 (1H,broad).
Example 228 (General procedure (C))
5-(2,3,6-Trichlorobenzylidene)thiazolidine-2,4-diαne
Figure imgf000168_0002
H PLC-MS (Method C): m/z: 309 (M+1); Rt.= 4.07 mirt
Example 229 (General procedure (C)) 5-(2,6-Dichlorobenzylidene)thiazolidine-2,4-dione
Figure imgf000168_0003
Mp. 152-1540C.
HPLC-MS (Method C): m/z: 274 (M+1), Rt.= 3.70 min
1H-NMR: (DMSO-de): 12.8 (1H, broad); 7.72 (1H,s); 7.60 (2H,d); 7.50 (1H,t). Example 230 (General procedure (C))
5-[1-(2,6-Dichloro-4-trifluoromethylphenyt>215-dimethyl-1H-pyrτol-3-ylmethy!ene]thiazolicline-
2,4-dione
Figure imgf000169_0001
5 HPLC-MS (Method C): m/z: 436 (M+1); Rt. 4.81 min
Example 231 (General procedure (C))
5-[1-(3,5-Dichlorophenyl)-5-(4-methanesuIfonylphenyl)-2-methyf-1H-pyrrol-3-ylmethylene]- thiazolidine-2,4-dione
Figure imgf000169_0002
HPLC-MS (Method C): m/z: 508 (M+1); Rt. = 4.31 min
Example 232 (General procedure (C))
5-[1-(2,5-Dimethoxypheny^5-(4-methanesuIfonylpheny!)-2-methyl-1H-pyrrol-3-ylmethylene]- 15 thiazolidine-2,4-dioπe
Figure imgf000169_0003
HPLC-MS (Method C): m/z: 499 (M+1); Rt. = 3.70 min
Exampte 233 (General procedure (C)) 20 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)-2,5-dimethylpyrrol-1-yl]benzoic acid
Figure imgf000170_0001
HPLC-MS (Method C): m/z:342 (M+ 1); Rt.= 3.19 min
Example 234 (General procedure (C)) 5-(4-Hydroxy-2,6-dimethoxybenzylidene)thiazolidine-2,4-dioπe
Figure imgf000170_0002
HPLC-MS (Method C): m/z:282( M+1); Rt = 2.56, mp=331-333 DC
Example 235 (General procedure (C))
5-(2,6-Dimethylbeπzylidene)thiazolidine-2,4-dione
Figure imgf000170_0003
M.p: 104-105 0C
HPLC-MS (Method C): m/z: 234 (M+1); Rt.= 3.58 min,
Example 236 (General procedure (C)) 5-(2,6-Dimethoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000170_0004
Mp: 241-242 CC HPLC-MS (Method C): m/z: 266 (M+1 ); Rt= 3.25 min; Example 237 (General procedure (C)) 5-[4-(2-Fluoro-6-nitrobenzyloxy)-2,6-dimethoxybenzylidene]thiazolidine-2,4-clione
Figure imgf000171_0001
Mp: 255-256 °C
HPLC-MS (Method C): m/z: 435 (M+1), Rt 4.13 min,
Example 238 (General procedure (C)) 5-Benzofuran-2-ylmethylenethiazolidine-2,4-dione
Figure imgf000171_0002
HPLC-MS (Method C): m/z:246 (M+1); Rt= 3.65 min, mp = 265-266 0C .
Example 239 (General procedure (C)) 5-[3-(4-Dimethylaminophenyl)allylidene]thiazolidine-2,4-dione
Figure imgf000171_0003
HPLC-MS (Method C): m/z:276(M+1); Rt = 3.63, mp = 259-263 0C
1H-NMR: {DMSO-dβ ) <J= 12.3 (1H,broad); 7.46 (2H,d); 7.39 (iH,d); 7.11 (1H,d); 6.69 (2H,d); 6.59 (1H, dd); 2.98 (3H1S).
Example 240 (General procedure (C)) 5-(2-Methyl-3-pheπylallylidene)thiazolidine-2,4-dϊone
Figure imgf000171_0004
Mp: 203-210 0C
HPLC-MS (Method C): m/z: 246 (M+1); Rt = 3.79 min. Example 241 (General procedure (C)) 5-{2-Chloro-3-phenylallylidene)thiazolidine-2,4-dione
Figure imgf000172_0001
Mp: 251-254 °C HPLC-MS (Method C): m/z: 266 (M+1 ; Rt = 3.90 min
Example 242 (General procedure (C)) 5-(2-Oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2)4-dione
Figure imgf000172_0002
Mp: 338-347 αC
HPLC-MS (Method C): m/z: 273 (M+1); Rt. = 2.59 min.
Example 243 (General procedure (C)) 5-(2,4,6-Tribromo-3-hydroxybenzylidene)thiazolidine-2,4-dione.
Figure imgf000172_0003
HPLC-MS (Method C): m/z: 459 (M+1);Rt.= 3.65 min.
Example 244 (General procedure (C)) 5-(5-Bromo-2-methyliπdol-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000172_0004
HPLC-MS (Method C): m/z: 339 (M+1 ); Rt = 3.37mirt.
Example 245 (General procedure (C)) 5-(7-Bromo-2-methylindol-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000173_0001
HPLC-MS (Method C): m/z: 319 (M+1): Rt = 3.48min.
Example 246 (General procedure (C)) 5-{6-Brornoindo!-3-ylrnethylene)thiazolidine-2,4-dione.
Figure imgf000173_0002
HPLC-MS (Method C): m/z: 325 (M+1); Rt = 3.54 min.
Example 247 (General procedure (C)) 5-(8-Methyl-2-oxo-1 ,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000173_0003
HPLC-MS (Method C): m/z: 287 (M+1); Rt = 2.86 min.
Example 248 (General procedure (C))
5-(6-Methoxy-2-oxo-1 ,2-dihydroquiπolin-3-ylmethylene)thlazolidine-2,4-dione.
Figure imgf000173_0004
HPLC-MS (Method C): m/z: 303 (M+1); Rt = 2.65 min.
Example 249 (General procedure (C))
5-Gufnoliπ-3-ylmethyteπetniazolidine-2,4-diσne.
Figure imgf000173_0005
HPLC-MS (Method C): m/z: 257 (M+1); Rt = 2.77 min. Example 250 (General procedure (C)) 5-(8-Hydroxyquinolin-2-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000174_0001
HPLC-MS (Method C): m/z: 273 (M+1); Rt = 3.44 min.
Example 251 (General procedure (C)) 5-Quinolin~8-ylmethy!enethiazolidine-2,4-dione.
Figure imgf000174_0002
HPLC-MS (Method C): m/z: 257 (M+1); Rt= 3.15 min.
Example 252 (General procedure (C)) S-ti-Bromo-€-methoxynaphthalen^-ylmethylene^hiazoliciine^^-dione.
Figure imgf000174_0003
HPLC-MS (Method C): m/z: 366 (M+1); Rt = 4.44 min.
Example 253 (General procedure (C)) 5-(6-Methyl-2-oxo-1,2-dihydroquinolin-3-ylmethylene)thiazolidine-2,4-dione.
Figure imgf000174_0004
HPLC-MS (Method C): m/z: 287 (M+1); Rt. = 2.89 min.
Example 254 (General procedure (D)) 5-(2,6-Dichloro-4-dibenzylaminobenzylidene)thiazolidine-2,4-ciione.
Figure imgf000174_0005
HPLC-MS (Method C): m/z: 469 (M+1 ); Rt = 5.35 min.
Example 255 (General Procedure (C))
7-(2,4-Dioxothia2θlidin-5-ylJdeπemethyJ)-4-me.hoxybenzαfuran-2-carboxylic acid
Figure imgf000175_0001
HPLC-MS (Method C): m/z: 320 (M+1); Rt = 2.71 mln.
Preparation of the intermediate, 7-forrnyl-4-methoxybenzofuran-2-carboxylic acid:
A mixture of 2-hydroxy-6-methoxybeπzaldehyde (6.4 g, 42 mmσl), ethyl brσmσacetate (14.2 mL, 128 mmol) and potassium carbonate (26 g, 185 mmol) was heated to 1300C. After 3 h the mixture was cooled to room temperature and acetone (100 mL) was added, the mixture was subsequently filtered and concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1 :4). This afforded 7.5 g (55%) of ethyl 4-methoxybenzofuran-2-carboxylate.
A solution of ethyl 4-methoxybenzofuran-2-carboxylate (6.9 g, 31.3 mmol) in dichloro- methane (70 ml) was cooled to 0 0C and a solution of titanium tetrachloride (13.08 g, 69 mmol) was added drop wise. After 10 minutes dichloromethoxymethane (3.958 g, 34 mmol) was added over 10 minutes. After addition, the mixture was warmed to room temperature for 18 hours and the mixture poured into hydrochloric acid (2N, 100 mL). The mixture was stirred for 0.5 hour and then extracted with a mixture of ethyl acetate and toluene (1:1). The organic phase was dried over Na2SO4 and concentrated in vacuo. The residue was purified by col¬ umn chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4). This afforded 5.8 g (80%) of ethyl 7-formyl-4-rrtethoxybenzofuran-2-carboxylate.
7-formyl-4-methoxybenzofuran-2-carboxylate (5.0 g, 21.5 mmol) and sodium carbonate (43 mmol) in water (100 mL) was refluxed until a clear solution appeared (about 0.5 hour). The solution was filtered and acidified to pH =1 with hydrochloric acid (2 N), the resulting product was filtered off and washed with ethyl acetate and ethanol and dried to afford 3.5 g (74%) of 7-formyl-4-methoxybβπzofuran-2-carboxylic acid as a solid. 1H NMR (DMSO-(Z6): δ = 10.20 (s, 1H) ; 8.07 (d, 1H) ; 7.70 (s, 1H) ; 7.17 (d, 1H) ; 4.08 (s, 3H).
Example 256 (General Procedure (C)) 5-(4-Methoxybenzofuran-7-ylmethy[ene)thiazolιdine-2,4-dione
Figure imgf000176_0001
HPLC-MS (Method C): mfc 267 (M+1); Rt = 3.30 min.
Preparation of the intermediate, 4-methoxybenzofuran-7-carbaldehyde:
A mixture of 7-forrnyl-4-methoxybenzofuran-2-carboxylic acid (3.0 g, 13.6 mmo!) and Cu (0.6 g, 9.44 mmol) in quinoline (6 mL) was refluxed. After 0.5 h the mixture was cooled to room temperature and water (100 mL) and hydrochloric acid (10 N, 20 mL) were added. The mix¬ ture was extracted with a mixture of ethyl acetate and toluene (1:1). filtered through celite and the organic layer separated and washed with a sodium carbonate solution, dried over Na2SO4 and concentrated in vacuo to afford 1.5 g crude product. Column chromatography SiO2, EtOAc/heptanes=1/4 gave 1.1 g (46%) of 4-methoxybenzofuran-7-carbaldehyde as a solid.
1H NMR (CDCI3): δ: 10.30 (s.1H) ; 7.85 (d,1H) ; 7.75 (d,1H) ; 6.98 (d,1H) ; 6.87 (d,1H) ; 4.10 (s,3H). HPLC-MS (Method C) :m/z: 177 (M+1); Rt. = 7.65 min.
Example 257 (General Procedure (C)) 5-(4-Hydroxybeπ2ofuran-7-ylmethylene)thiazolidine-2,4-dioπe
Figure imgf000176_0002
HPLC-MS (Method C): m/z: = 262 (M+1); Rt 2.45 min.
Preparation of the intermediate, 4-hydroxybenzofuran-7-carbaldehyde A mixture of 4-methoxybenzofuran~7-carbaldehyde (1.6 gf 9.1 mmol) and pyridine hydrochlo¬ ride (4.8 g, 41.7mmol) in qufnoline (8 mL) was reffuxed. After 8 h the mixture was cooled to room temperature and poured into water (100 mL) and hydrochloric acid (2 N) was added to pH = 2. The mixture was extracted with a mixture of ethyl acetate and toluene (1:1), washed with a sodium carbonate solution, dried with Na2SO4 and concentrated in vacuo to afford 0.8 g crude product. This was purified by column chromatography on silica gel, eluting with a mixture of ethyl acetate and heptane (1:3). This afforded 250 mg of 4-hydroxybenzofuran-7- carbaldehyde as a solid.
1H NMR (DMSO-Of5): 6 = 11.35 (sf broad, 1H) ; 10.15 (s, 1H) ; 8.05 (df 1H) ; 7.75 (d, 1H) ; 7.10 (d, 1H); 6.83 (d, 1H). HPLC-MS (Method C): m/z: 163 (M+1); Rt. = 6.36 min.
Example 258 (Genera! Procedure (C))
5-(5-Bromo-2,3-dihydrobenzofuran-7-ylmethylene)thiazolidine-2,4-dione
Figure imgf000177_0001
HPLC-MS (Method C): m/z: 328 (M+1); Rt = 3.66 min.
Preparation of the intermediate, 5-bromo-2,3-dihydrobenzofuran-7-carbaldehyde:
To a cooled (150C) stirred mixture dihydrobenzofuran (50.9 g, 0.424 mol) in acetic acid (500 mL), a solution of bromine (65.5 mL, 1.27 mol) in acetic acid (200 mL) was added drop wise over 1 hour. After stirring for 18 hours, a mixture of Na2S2Os (150 g) in water (250 mL) was added carefully, and the mixture was concentrated in vacuo. Water (200 mL) was added and the mixture was extracted with ethyl acetate containing 10% heptane, dried over Na2SO4 and concentrated in vacuo to give crude 5,7-dibromo-2,3-dihydrobenzofuran which was used as such for the following reaction steps. To a cooled solution (-780C) of crude 5,7-dibromo-2,3- dihydrobenzofuran (50.7 g, 0.182 mol) in THF (375 mL) a solution of n-BuLi (2.5 M, 80 mL, 0.200 mol) in hexane was added. After addition, the mixture was stirred for 20 min. DMF (16 mL) was then added drop wise at -780C. After addition, the mixture was stirred at room tem¬ perature for 3 h and then the mixture was poured into a mixture of ice water, (500 mL) and hydrochloric acid (10 N, 40 mL) and extracted with toluene, dried over Na2SO4 and concen¬ trated in vacuo. Column chromatography on silica gel eluting with a mixture of ethyl acetate and heptane (1:4) afforede 23 g of 5-bromo-2,3-dihydrobenzofuran-7-carbaldehyde as a solid.
1H NMR (CDCI3): «5:10.18 (s,1H) ; 7.75 (d,1H) ;7.55 (d.1H) ; 4.80 (t,2H) ; 3.28 (t,2H).
Example 259 (General Procedure (C)) 5-(4-Cyc(ohexyIbenzy[idene)thiazolidiπe-2,4-dione
Figure imgf000178_0001
HPLC-MS (Method C): m/z: 288 (M+1); Rt = 5.03 min.
Preparation of the intermediate, 4-cyclohexylbenzaldehyde:
This compound was synthesized according to a modified literature procedure (J. Org. Chem., 37, No.24, (1972), 3972-3973). Cyclohexylbenzene (112.5 g, 0.702 mol) and hexamethylenetetramine (99.3 g, 0.708 mol) were mixed in TFA (375 mL). The mixture was stirred under nitrogen at 900C for 3 days. Af¬ ter cooling to room temperature the red-brown mixture was poured into ice-water (3600 ml) and stirred for 1 hour. The solution was neutralized with Na2CO3 (2 M solution in water) and extracted with dichloromethane (2.5 L). The organic phase was dried (Na2SO4) and the sol- vent was removed in vacuo. The remaining red-brown oil was purified by fractional distillation to afford the title compound (51 g, 39%).
1H NMR (CDCI3): £9.96 (s, 1H), 7.80 (d, 2H), 7.35 (d, 2H). 2.58 (m, 1H), 1.94-1.70 (m, 5 H), 1.51-1.17 (m, 5H)
Other ligands of the invention include
3',5'-Dichloro-4"-(2,4-dioxothiazolidin-5-ylidenemethyl)biphenyl-4-carboxylic acid:
Figure imgf000178_0002
Example 260 (General procedure (C)) 5-(1-Bromo-6-hydroxynaphthalen-2-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000179_0001
HPLC-MS (Method C): m/z = 350 (M+1 ); Rt. = 3.45 min.
Example 261 (General procedure (C))
5-[4-(2-Bromoethoxy)-naphthalen-1-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000179_0002
HPLC-MS (Method C): m/z = 380 (M+1 ); Rt = 3.52 min.
Example 262 (General procedure (C))
5-(2-Methyl-5-nitro-1H-indol-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000179_0003
HPLC-MS (Method C): m/z = 304 (M+1 ); Rt = 2.95 min.
Example 263 (General procedure (C)) 5-(4-Naphthalen-2-yl-thiazol-2-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000180_0001
HPLC-MS (Method C): m/z = 339 (M+1); Rt.= 4.498 min.
Example 264 (General procedure (C)) 5-[4-(4-Methoxy-naphthalen-1-yl)-thiazol-2-ylmethylene]-thia2olidine-2,4-dione
Figure imgf000180_0002
HPLC-MS (Method C): m/z = 369 (M+1); Rt= 4.456 mm.
Example 265 (General procedure (C)) 5-(2-Pyridin-4-yl-1 H-indol-3-yImethy!ene)-thiazolidine-2,4-dione
Figure imgf000180_0003
HPLC-MS (Method C): m/z = 322 (M+1 ); Rt. = 2.307 min.
Example 266 (General procedure (C)) 5-[5-{4-Chlorophenyl)-1H-pyrazol-4-ylmethy!ene]-thiazolidine-2,4-dione
Figure imgf000181_0001
HPLC-MS (Method C): m/z = 306 (M+1); Rt.= 3.60 min.
Example 267 (General procedure (C)) 5-[5-(2,5-Dimethylphenyl)-1H-pyrazol-4-ylmethyleπe]-thiazolidine-2,4-dione
Figure imgf000181_0002
HPLC-MS (Method C): m/z = 300 (M+1); Rt. = 3.063 min.
Example 268 (General procedure (C))
5-(2-Phenyl-benzo[d]imidazo[2,1-b]thiazol-3-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000181_0003
HPLC-MS (Method C): m/z = 378 (M+1); Rt = 3.90 min.
Example 269 (General procedure (C))
N-{4-[2-(2,4-Dioxothiazolidin-5-ylidenemethyl)-phenoxy]-phenyl}-acetamide
Figure imgf000182_0001
HPLC-MS (Method C): m/z = 355 (M+1); Rt 3.33 min.
Example 270 (General procedure (C)) 5-(2-Phenyl-imidazo[1,2-a]pyridin-3-ylmethyleneHhiazolidine-214-dione
Figure imgf000182_0002
HPLC-MS (Method C): m/z = 322 (M+1); Rt. = 2,78 min.
Example 271 (General procedure (C)) 5-(2-Naphthalen-2-yl-imidazo[1 ,2-a]pyridin-3-ylmethylene)-thiazolidine-2,4-dioπe
Figure imgf000182_0003
HPLC-MS (Method C): m/z = 372 (M+1); Rt. = 2.78 min.
Example 272 (General procedure (C)) 5-[6-Bromo-2-(3-methoxyphenyl)-imidazo[1 ,2-a]pyridin-3-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000183_0001
HPLC-MS (Method C): m/z = 431 (M+1); Rt.= 3.30 min.
Example 273 (General procedure (C)) 5-(1,2,3,4-Tetrahydrophenanthren-9-ylmethylene)thiazolidine-2r4-dione
Figure imgf000183_0002
HPLC-MS (Method C): m/z = 310 (M+1 }; Rt.= 4.97 min.
Example 274 (General procedure (C)) 5-(3,5,5,8.8-Pentamethyl-51617,8-tetrahydro-naphthalen-2-ylmethylene)thia2olidine-2,4-dione
Figure imgf000183_0003
HPLC-MS (Method C): m/z = 330 (M+1); Rt.= 5.33 min.
Example 275 (General procedure (C)) 5-[6-(2,4-Dichloro-phenyl)-imidazo[2,1-b]thiazol-5-ylmethylene]-thiazolidine-2,4-dione
Figure imgf000184_0001
HPLC-MS (Method C): m/z = 396 (M+1); Rt. = 3.82 min.
Example 276 (General procedure (C)) 5-(5-Bromobenzofuraπ-7-ylmethylene)-thiazolidine-2,4-dione
Figure imgf000184_0002
HPLC-MS (Method C): m/z = 324 (M+1); Rt. = 3.82 min.
Example 277 (General procedure (C)) 4-[3-(2,4-Dioxothiazolidin-5-ylfdenemethyi)-1,4-dimethylcarbazol-9-ylmethyl]-benzoic acid
Figure imgf000184_0003
HPLC-MS (Method C): m/z = 457 (M+1); Rt = 4,23 min.
Preparation of intermediary aldehyde: 1,4 Dimethylcarbazαl-3-carbaldehyde (0.68 g, 3.08 mmol) was dissolved in dry DMF (15 mL), NaH (diethyl ether washed) (0.162 g, 6.7 mol) was slowly added under nitrogen and the mix¬ ture was stirred for 1 hour at room temperature. 4-Bromomethylbenzoic acid (0.73 g, 3.4 mmol) was slowly added and the resulting slurry was heated to 400C for 16 hours. Water (5 rnL) and hydrochloric acid (6N, 3 mL) were added. After stirring for 20 min at room tempera¬ ture, the precipitate was filtered off and washed twice with acetone to afford after drying 0.38 g (34%) of 4-(3-formy!-1 ,4-dimethylcarbazol-9-ylmethyl)beπzofc acid.
HPLC-MS (Method C) : m/z = 358 <M+1), RT. = 4.15 min.
Example 278 (General procedure (C)) 4-[7-(2,4-Dioxothiazolidin-5-ylidenemethyl)-benzofuran-5-yl]-benzoic acid
Figure imgf000185_0001
Starting aldehyde commercially available (Syncom BV, NL) HPLC-MS (Method C): m/z = 366 (M+1); Rt. = 3.37 min.
Example 279 (General procedure (C)) 4_[4-(2,4-Diσxothiazolidin-5-ylideπemethyl)-2-nitrophenoxy]-benzαic acid methyl ester
Figure imgf000185_0002
HPLC-MS (Method C): m/z = 401 (M+1); Rt. = 4.08 min.
Example 280 (General procedure (C)) S'.δ'-Dichloro-^-t∑^-dioxothiazolidin-δ-ylidenemethylJ-biphenyW-carboxylic acid
Figure imgf000186_0001
Starting aldehyde commercially available (Syncom BV, NL)
HPLC-MS {Method C): m/z = 394 (M+1); Rt. = 3.71 min.
Example 281 (General procedure (C))
Figure imgf000186_0002
HPLC-MS (Method C}: m/z = 232( M+1 ); Rt = 3.6 min.
Example 282
5~(2-Methyl-1H-indol-3-ylmethyl)-thiazolidine-2,4-dione
Figure imgf000186_0003
5-(2-Methyl-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione (prepared as described in example 187, 1.5 g. 5.8 mmol) was dissolved in pyridine (20 mL) and THF (50 mL), LiBH4 (2 M in THF, 23.2 mmol) was slowly added with a syringe under cooling on ice. The mixture was heated to 85 0C for 2 days. After cooling, the mixture was acidified with concentrated hydro¬ chloric acid to pH 1. The aquous layer was extracted 3 times with ethyl acetate, dried with MgSO4 treated with activated carbon, filtered and the resulting filtrate was evaporated in vacuo to give 1.3 g (88%) of the title compound.
HPLC-MS (Method C): m/z = 261 (M+1); Rt. = 3.00 min. Example 283 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]buty ric acid
Figure imgf000187_0001
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphtha!en-1-yloxy]butyric acid (4.98 g, 13.9 mmol, prepared as described in example 469) was dissolved in dry THF (50 mL) and added dry pyridine (50 mL) and, in portions, lithium borohydride (2.0 M1 in THF, 14 mL). The result¬ ing slurry was refluxed under nitrogen for 16 hours, added (after cooling) more lithium boro¬ hydride (2.0 M1 in THF, 7 mL). The resulting mixture was refluxed under nitrogen for 16 hours. The mixture was cooled and added more lithium borohydride (2.0 M, in THF, 5 mL). The resulting mixture was refluxed under nitrogen for 16 hours. After cooling to 5 °C, the mix¬ ture was added water (300 mL) and hydrochloric acid (150 mL). The solid was isolated by filtration, washed with water (3 x 500 mL) and dried. Recrystallization from acetonitrile (500 mL) afforded2.5 g of the title compound.
1H-NMR (DMSO-dβ, selected peaks): δ = 3.42 (1H, dd), 3.90 (114, dd), 4.16 (2H, T), 4.95 (1 H, dd), 6.92 (1 H, d), 7.31 (1 H, d). 7.54 (1 H, t), 7.62 (1H, t). 8.02 (1H. d), 8.23 (1H, d), 12.1 (1H, bs), 12.2 (1 H, bs). HPLC-MS (Method C): m/z = 382 (M+23); Rt * 3.23 miπ.
Example 284 5-Naphthalen-1-y!methylthiazolidine-2,4-dione
Figure imgf000187_0002
5-Naphthalen-1-ylmethylenethiazolidine-2,4-dione (1.08 g, 4.2 mmol, prepared as described in example 68) was dissolved in dry THF (15 mL) and added dry pyridine (15 mL) and, in portions, lithium borohydride (2.0 M, in THF, 4,6 mL). The resulting mixture was refluxed un¬ der nitrogen for 16 hours. After cooling to 5 0C, the mixture was added water (100 mL), and, in portions, concentrated hydrochloric acid (40 mL). More water (100 mL) was added, and the mixture was extracted with ethyl acetate (200 mL). The organic phase was washed with water (3 x 100 mL), dried and concentrated in vacuo. The residue was dissolved in ethyl ace¬ tate (50 mL) added activated carbon, filtered and concentrated in vacuo and dried to afford 0.82 g (75%) of the title compound.
1H-NMR (DMSO-Cf6): δ = 3.54 (1H, dd), 3.98 (1 H, dd), 5.00 (1H, dd), 7.4-7.6 (4H, m), 7.87 (1H, d), 7.96 (1H1 d), 8.11 (1H, d), 12.2 (1H. bs). HPLC-MS (Method C): m/z = 258 (M+1 ); Rt = 3,638 min.
The following preferred compounds of the invention may be prepared according to proce¬ dures similar to those described in the three examples above:
Figure imgf000188_0001
Figure imgf000189_0001
Figure imgf000190_0001
Figure imgf000191_0001
Figure imgf000192_0001
Figure imgf000193_0001
Figure imgf000194_0001
Figure imgf000195_0001
Figure imgf000196_0001
Figure imgf000197_0001
Figure imgf000198_0001
Figure imgf000199_0001
Figure imgf000200_0003
The following compounds are commercially available and may be prepared using general procedures (B) and / or (C). Example 380 5-(5-Bromo-1 H-indol-3-ylmethylene)thiazolidine-2,4-dione
Figure imgf000200_0001
Example 381
5-Pyridin-4-ylmethylenethiazolidine-2,4-dione
Figure imgf000200_0002
Example 382 5-{3-Bromo-4-methoxybenzylidene)thiazolidine-2,4-dione
Figure imgf000201_0001
Example 383 5-(3-Nitrobenzylideπe)thiazolidine-2,4-dione
Figure imgf000201_0002
Example 384 5-Cyclohexylidene-1 ,3-thiazo1idine-2,4-dione
Figure imgf000201_0003
Example 385 5-(3,4-Dihydroxybenzylidene)thiazo!idine-2,4-dione
Figure imgf000201_0004
Example 386 5-(3-Ethoxy-4-hydroxybenzyIidene)thiazolidine-2,4-dione
Figure imgf000201_0005
Example 387 5-(4-Hydroxy-3-methoxy-5-nitrobenzylidene)thfazoJidine-2,4-dione
Figure imgf000202_0001
Example 388
5-(3-Etnoxy-4-hydra5<yberi∑ylidene)thiazoticJine-2,4-dSone
Figure imgf000202_0002
Example 389 5-(4-Hydroxy-3,5-ditnethoxyfaen2ylidene)thiazolidinβ-2,4-dioπe
Figure imgf000202_0003
Exampfe 390
5-(3-Bromo-5-ethoxy-4-hydroxyben2ylidene)thia2θltdine~2,4-dione
Figure imgf000202_0004
Example 391
5-(3-Ethoxy-4-hydroxy-5-πitrobenzylidene)thia2olidine-2,4-diorie
Figure imgf000202_0005
Example 392
Figure imgf000203_0001
Example 393
Figure imgf000203_0002
Example 394
Figure imgf000203_0003
Example 395
Figure imgf000203_0004
Example 396
Figure imgf000203_0005
Example 397
Figure imgf000204_0001
Example 398
Figure imgf000204_0002
Example 399
Figure imgf000204_0003
Example 400
Figure imgf000204_0004
Example 401
Figure imgf000204_0005
Example 402
Figure imgf000205_0001
Figure imgf000205_0002
Example 405
5-{3-Hydroxy>5-methyl-phenylamino)-thiazolidine-2,4-dione
Figure imgf000205_0003
Example 406
Figure imgf000206_0001
Example 407
Figure imgf000206_0002
Example 409
Figure imgf000206_0003
Example 410
Figure imgf000207_0001
Example 411
Figure imgf000207_0002
Example 412
Figure imgf000207_0003
Example 413
Figure imgf000207_0004
Example 414
Figure imgf000207_0005
Example 415
Figure imgf000208_0001
Example 416
Figure imgf000208_0002
Example 417
Figure imgf000208_0003
Example 418
Figure imgf000208_0004
Example 420
Figure imgf000209_0001
Example 421
Figure imgf000209_0002
Example 422
Figure imgf000209_0003
Example 423
Figure imgf000209_0004
Example 424
Figure imgf000209_0005
Example 425
Figure imgf000210_0001
Example 426
Figure imgf000210_0002
Example 427
Figure imgf000210_0003
Example 428
Figure imgf000210_0004
Example 430
Figure imgf000211_0001
Example 431 5-(4-D(ethylamino-2-methoxy-ben2ylidene)-irπidarolidine-2,4-dione
Figure imgf000211_0002
Example 432
Figure imgf000211_0003
Example 433
Figure imgf000211_0004
Example 434
Figure imgf000212_0001
Example 435
Figure imgf000212_0002
Example 436
Figure imgf000212_0003
Example 437
Figure imgf000212_0004
Example 438
Figure imgf000213_0001
Example 439
Figure imgf000213_0002
Example 440
Figure imgf000213_0003
Example 441
Figure imgf000214_0001
Figure imgf000214_0002
Example 443
Figure imgf000214_0003
Example 444
Figure imgf000214_0004
Example 445
Figure imgf000215_0002
Example 447
Figure imgf000215_0003
Figure imgf000215_0004
Example 449
Figure imgf000215_0005
Example 450
Figure imgf000216_0001
Example 451
Figure imgf000216_0002
Example 452
Figure imgf000216_0003
Example 453
Figure imgf000216_0004
Example 454 5-(4~Diethy}amino-benzy}idene)-2-imino-thiazo}idin-4-one
Figure imgf000217_0001
Example 455
Figure imgf000217_0002
Example 456
Figure imgf000217_0003
Example 457
Figure imgf000217_0004
Example 458
Figure imgf000218_0001
Figure imgf000218_0002
General procedure (D) for preparation of compounds of general formula
I3:
Figure imgf000218_0003
wherein X. Y, and R3 are as defined above, n is 1 or 3-20.
E is arylene or heterarylene (including up to four optional substituents, R13, R14, R15, and R15A as defined above),
R' is a standard carboxylic acid protecting group, such as Ci-Cβ-atkyl or benzyl and Lea is a leaving group, such as chloro, bromo, iodo, methanesulfonyloxy, toluenesulfonyloxy or the like. Step 1 is an alkylation of a phenol moiety. The reaction is preformed by reacting R10-C(=O)- E-OH with an ω-brorno-alkane-carboxylic acid ester (or a synthetic equivalent) in the pres¬ ence of a base such as sodium or potassium carbonate, sodium or potassium hydroxide, so- dium hydride, sodium or potassium alkoxide in a solvent, such as DMF, NMP, DMSO, ace¬ tone, acetonitrile, ethyl acetate or isopropyl acetate. The reaction is performed at 20 - 160 0C, usually at room temperature, but when the phenol moiety has one or more substituents heating to 500C or more can be beneficial, especially when the substituents are in the ortho position relatively to the phenol. This will readily be recognised by those skilled in the art.
Step 2 is a hydrolysis of the product from step 1.
Step 3 is similar to general procedure (B) and (C).
This general procedure (D) is further illustrated in the following examples:
Example 460 (General procedure (D)) 4-[4-(2,4-Dioxothiazolidin-5-ylideπemethyl)phenoxy]butyric acid
Figure imgf000219_0001
Step i:
A mixture of 4-hydroxybenzaldehyde (9.21 g, 75 mmol), potassium carbonate (56 g, 410 mmol) and 4-bromobutyric acid ethyl ester (12.9 ml_, 90 mmol) in A/./V-dimethylformamide (250 mL) was stirred vigorously for 16 hours at room temperature. The mixture was filtered and concentrated in vacuo to afford 19.6 g (100%) of 4-(4-formylphenoxy)butyric acid ethyl ester as an oil. 1H-NMR (DMSO-(Z6): δ 1.21 (3H, t), 2.05 (2H, p), 2.49 (2H, t), 4.12 (4H, m), 7.13 (2H1 d), 7.87 (2H, d), 9.90 (1H1 s). HPLC-MS (Method A): m/z = 237 (M+1); R, = 3.46 min.
Step 2: 4-(4-Formylphenoxy)butyric acid ethyl ester (19.6 g, 75 mmol) was dissolved in methanol (250 mL) and 1N sodium hydroxide (100 mL) was added and the resulting mixture was stirred at room temperature for 16 hours. The organic solvent was evaporated in vacuo (40 0C, 120 mBar) and the residue was acidified with 1N hydrochloric acid (110 mL). The mixture was filtered and washed with water and dried in vacuo to afford 14.3 g (91%) 4-(4- formylphenoxy)butyric acid as a solid. 1H-NMR (DMSOcfe): δ 1.99 (2H, p), 2.42 (2H, t), 4.13 (2H, t). 7.14 (2H, d), 7.88 (2H, d), 9.90 <1H, s), 12.2 (1H, bs). HPLC-MS (Method A): mfe = 209 (M+1); R, = 2.i9 min.
Step 3:
Thiazolidϊne-2.4-dione (3.55 g, 27.6 mmol), 4-(4-formylphenoxy)butyric acid (5.74 g, 27.6 mmol), anhydrous sodium acetate (11.3 g, 138 mmol) and acetic acid (100 mL) was refluxed for 16 h. After cooling, the mixture was filtered and washed with acetic acid and water. Drying in vacuo afforded 2.74 g (32%) of 4-[4-(2,4-dioxothiazolidin-5-yIidenemethyl)phenoxy]butyric acid as a solid.
1H-NMR (DMSO-Of6): <?1.97 (2H. p), 2.40 (2H, t), 4.07 (2H1 1), 7.08 (2H, d). 7.56 (2H, d). 7.77 (1H, s), 12.2 (1H, bs), 12.5 (1H1 bs); HPLC-MS (Method A): m/z: 308 (M+1); Rt = 2.89 min.
Example 461 (General procedure (D))
[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)pheπoxy]acetic acid
Figure imgf000220_0001
Step 3:
Thiazolidine-2,4-dtone (3.9 g, 33 mmol), 3-formylphenoxyacetic acid (6.0 g, 33 mmol), anhy- drous sodium acetate (13.6 g, 165 mmol) and acetic acid (100 mL) was refluxed for 16 h. Af¬ ter cooling, the mixture was filtered and washed with acetic acid and water. Drying in vacuo afforded 5.13 g (56%) of [3-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid as a solid.
1H-NMR (DMSO-Cf6): δ 4.69 (2H, s), 6.95 (1Hr dd). 7.09 (1H, t), 7.15 (1H1 d). 7.39 (1H, t),7.53 (1 H, s); HPLC-MS (Method A): m/z = 280 (M+1) (poor ionisation); R, = 2.49 min.
The compounds in the following examples were similarly prepared.
Exampfe 462 (General procedure (D)) S-^-^^-Dioxothiazolidin-S-ylideπemethylJphenyϊlacrylic acid
Figure imgf000221_0001
1H-NMR (DMSO-d6): «?6.63 (1H1 d), 7.59-7.64 (3H, m), 7.77 (1H, s), 7.83 (2H, m).
Example 463 (General procedure (D)) [4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetic acid
Figure imgf000221_0002
Triethylamine salt: 1H-NMR (DMSO-Cf6): δ 4.27 (2Hr s), 6.90 (2H. d). 7.26 (1H, s), 7.40 (2H, d).
Example 464 (General procedure (D)) 4-(2,4-Dioxothiazolidin-5-ylideπemethyl)benzoic acid
Figure imgf000221_0003
Example 465 (General procedure (D))
3-(2,4-Dioxothiazolidin-5-ylideπemethyI)benzoicacid
Figure imgf000221_0004
1H-NMR (DMSO-Ck): δ 7.57 (1H, s), 7.60 (1H, t), 7.79 (1H, dt), 7.92 (1H, dt), 8.14 (1H, t).
Example 466 (General procedure (D))
4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000222_0001
1H-NMR (DMSO-cfe): δ 2.00 (2H, p), 2.45 (2H, t), 4.17 (2H1 1), 7.31 (1 H, d). 7.54 (1H1 dd), 7.69 (1H, d), 7.74 (1H, s), 12.2 (1H, bs), 12.6 (1H. bs). HPLC-MS (Method A): m/z: 364 (M+23); Rt = 3.19 min.
Example 467 (General procedure (D)) 4-t2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000222_0002
1H-NMR (DMSO-Ck): $ 1.99 (2H, p), 2.46 (2H. t), 4.17 (2H, t). 7.28 (1H, d). 7.57 (1H, dd). 7.25 (1H1 s), 7.85 (1H, d), 12.2 (1H1 bs). 12.6 (1H1 bs). HPLC-MS (Method A): m/z: 410 (M+23); Rt = 3.35 min.
Example 468 (General procedure (D)) 4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000222_0003
1H-NMR (DMSO-dβ): δ 1.99 (2H. p). 2.45 (2H1 1), 4.18 (2H, t), 7.28 (1H. d). 7.55 (1H. dd), 7.60 (1H, s). 7.86 (1H, d), 12.2 (1H1 bs), 13.8 (1H, bs). HPLC-MS (Method A): m/z: 424 (M+23); Rt = 3.84 min. HPLC-MS (Method A): m/z: 424 (M+23); Rt = 3,84 min
Example 469 (General procedure (D)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid
Figure imgf000222_0004
1H-NMR (DMSO-ctø: δ 2.12 (2H1 p), 2.5 (below DMSO), 4.28 (2H, t), 7.12 (1H, d), 7.6-7.7 (3H, m). 8.12 (1H, d), 8.31 (1H, d), 8.39 (1H, s), 12.2 (1H, bs), 12.6 (1H, bs). HPLC-MS (Method A): m/z: 380 (M+23); Rt = 3.76 min.
Example 470 (General procedure (D))
5-[4-(2,4-Dioxothiazolidin-5-ylideπemethyl)πaphtha!eπ-1-yloxy]pentanoic acid
Figure imgf000223_0001
HPLC-MS (Method A): m/z: 394 (M+23); Rt = 3.62 min.
1H-NMR (DMSO-dβ): δ 1.78 (2H1 m), 1.90 (2H, m). 2.38 (2H. t).4.27 (2H, t), 7.16 (1H. d), 7.6-7.75 (3H, m), 8.13 (1H1 d), 8.28 (1H1 d). 8.39 (1H. s), 12.1 (1H1 bs), 12.6 (1H. bs).
Example 471 S-^-Bromo^Z^-dioxothiazolidin-S-ylidenemethyOnaphthalen-i-yloxyjpentanoic acid.
Figure imgf000223_0002
5-[4-(2,4'Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]pentanoic acid (example 470, 185 mg, 0.5 mmol) was treated with an equimolar amount of bromine in acetic acid (10 mL). Stirring at RT for 14 days followed by evaporation to dryness afforded a mixture of the bro- minated compound and unchanged starting material. Purification by preparative HPLC on a C18 column using acetonitrile and water as eluent afforded 8 mg of the title compound.
HPLC-MS (Method C): m/z: 473 (M+23), Rt. = 3.77 min
Example 472 4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethy!)naphthalen-1-yloxy]butyric acid.
Figure imgf000223_0003
Starting with 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyI)-naphthalen-1-yloxy]-butyric acid (ex¬ ample 469, 0.5 mmol) using the same method as in example 471 afforded 66 mg of the title compound,
HPLC-MS {Method C): m/z: 459 (M+23) ; Rt. = 3.59 min.
Example 473 (General procedure (D))
[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)pheπoxy]acetic acid
Figure imgf000224_0001
1H-NMR (DMSO-CZ6): δ 4.90 (2H1 s), 7.12 (1H, d), 7.52 (1H, dd). 7.65 (1H, s) 7.84 (1H, d).HPLC-MS (Method A): m/z: not observed; Rt = 2.89 min.
Example 474 (General procedure (D)) 4-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyric acid
Figure imgf000224_0002
1H-NMR (DMSO-tfβ): δ 1.98 (2H, p), 2.42 (2H, t), 4.04 (2H. t). 7.05 (1H. dd), 7.15 (2H, m), 7.45 (1H, t), 7.77 (1H, s), 12.1 (1H, bs), 12.6 (1H1 bs). HPLC-MS (Method A): m/z: 330 (M+23); Rt = 3.05 min.
Example 475 (General procedure (D))
[4-(2,4-Dioxothiazolidin-5-ylϊdenemethyI)-3-methoxyphenoxy]acetic acid
Figure imgf000224_0003
HPLC-MS (Method B): m/z: 310 (M+1); Rt = 3,43 min.
Example 476 (Genera! procedure (D)) [4-{214-Dioxothiazolidin-5-yIfdenemethyl)naρhthalen-1-yloxy]acetic acid
Figure imgf000225_0001
HPLC-MS (Method A): m/z: 330 (M+1); Rt = 3.25 min.
Example 477 (General procedure (D)) 8-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalene-1 -carboxylic acid
Figure imgf000225_0002
HPLC-MS (Method A): m/z: 299 (M+1 ); Rt = 2,49 min.
Example 478 (General procedure (D)) [3-(2,4-Dioxothiazolidiπ-5-ylidenemethyl)indol-1 -yl]acetic acid
Figure imgf000225_0003
HPLC-MS (Method A): m/z: 303 (M+1); Rt = 2.90 min.
Preparation of starting material: 3-Formyliπdol (10 g, 69 mmol) was dissolved in W.W-dimethylfαrmamide (100 mL) and under an atmosphere of nitrogenand with external cooling, keeping the temperature below 150C, sodium hydride (60% in mineral oil, 3.0 g, 76 mmol) was added in portions. Then a solution of ethyl bromoacetate (8.4 mL, 76 mmol) in Λ/,Λ/-dimethy|formamide (15 mL) was added dropwise over 30 minutes and the resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo and the residue was partitioned between wa¬ ter (300 mL) and ethyl acetate (2 x 150 mL). The combined organic extracts were washed with a saturated aqueous solution of ammonium chloride (100 mL), dried (MgSO4) and con¬ centrated in vacuo to afford 15.9 g (quant.) of (3-formyliπdol-1-yl)acetic acid ethyl ester as an oil. 1H-NMR (CDCI3): δH = 1-30 (3H, t), 4.23 (2H, q), 4.90 (2H, s), 7.3 (3H1 m), 7.77 (1H, s), 8.32 (1H, d). 10.0 (1H, s).
(3-Foππylindol-1-yl)acetic acid ethyl ester (15.9 g 69 mmol) was dissolved in 1,4-dioxane (100 mL) and 1N sodium hydroxide (10 mL) was added and the resulting mixture was stirred at room temperature for 4 days. Water (500 mL) was added and the mixture was washed with diethyl ether (150 mL). The aqueous phase was acidified with SN hydrochloric acid and extracted with ethyl acetate (250 + 150 mL). The combined organic extracts were dried (MgSO4) and concentrated in vacuo to afford 10.3 g (73%) of (3-formylindol-1 -yl)acetic acid as a solid.
1H-NMR (DMSO-Oe): δH = 5.20 (2H, s), 7.3 (2H, m), 7.55 (1H, d), 8.12 (1H, d), 8.30 (1H, s), 9.95 (1H, s). 13.3 (1 H, bs).
Example 479 (General procedure (D)) 3-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-yl]propionic acid
Figure imgf000226_0001
HPLC-MS (Method A): m/z: 317 (M+1); Rt = 3.08 min.
Preparation of starting material:
A mixture of 3-formylindol (10 g. 69 mmol), ethyl 3-bromopropionate (10.5 mL, 83 mmol) and potassium carbonate (28.5 g, 207 mmol) and acetonitrile (100 mL) was stirred vigorously at refux temperature for 2 days. After cooling, the mixture was filtered and the filtrate was con- centrated in vacuo to afford 17.5 g (quant.) of 3-(3-formylindol-1-yl)propionic acid ethyl ester as a solid.
1H-NMR (DMSO-Cf8): δH = 1.10 (3H, t), 2.94 (2H, t), 4.02 (2H. q), 4.55 (2H, t)t 7.3 (2H, m), 7.67 (1H, d), 8.12 (1H, d), 8.30 (1H, s), 9.90 (1H1 s). 3-(3-Formylindol-1-yl)propionic acid ethyl ester (17.5 g 69 mmol) was hydrolysed as de¬ scribed above to afford 12.5 g (83%) of 3-(3-formyliπdoI-1-yl)propionic acid as a solid.
1H-NMR (DMSO-tfβ): δH = 2.87 (2H, t), 4.50 (2H, t), 7.3 (2H, m), 7.68 (1H, d), 8.12 (1H, d), 8.31 (1 H, s), 9.95 (1 H1 s), 12.5 (1 H, bs).
Example 480 (General procedure (D))
{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzylideπe]-4-oxo-2-thioxothiazolidin-3-yl}acetic acid
Figure imgf000227_0001
HPLC-MS (Method A): m/z: 429 (M+23); Rt = 3.89 min.
Example 481 (General procedure (D)) 6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naprithalen-2-yloxyoctanoic acid
Figure imgf000227_0002
HPLC-MS (Method C): m/z: 436 (M+23); Rt.= 4.36 min
The intermediate aldehyde for this compound was prepared by a slightly modified procedure: 6-Hydroxynaphthalene-2-carba!dehyde (1.0 g, 5.8 mmol) was dissolved in DMF (10 mL) and sodium hydride 60% (278 mg) was added and the mixture stirred at RT for 15 min. 8- Bromooctanoic acid (0.37 g, 1.7 mmol) was converted to the sodium salt by addition of so¬ dium hydride 60% and added to an aliquot (2.5 mL) of the above naphtholate solution and the resulting mixture was stirred at RT for 16 hours. Aqueous acetic acid (10 %) was added and the mixture was extracted 3 times with diethyl ether. The combined organic phases were dried with MgSO4 and evaporated to dryness affording 300 mg of 8-(6-formylnaphthalen-2- yloxy)octanoic acid. HPLC-MS (Method C): m/z 315 (M+1); Rt. = 4.24 min. Example 482 (General procedure (D))
12-[6-(2,4-Dioxothiazolidin-5-ylidenernethyl)naphthalen-2-yfoxy]dodecaπoic acid .
Figure imgf000228_0001
HPLC-MS (Method C): m/z: 492 (M+23); Rt = 5.3 rnin.
The intermediate aldehyde was prepared similarly as described in example 481.
Example 483 (General procedure (D))
11 -[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]undecanoic acid.
Figure imgf000228_0002
HPLC-MS (Method C): m/z:478 (M+23); Rt.= 5.17 min.
The intermediate aldehyde was prepared similarly as described in example 481.
Example 484 (General procedure (D)) iδ-ie^Λ-Dioxothiazolidin-δ-ylidenemethylJnaphthal'en^-yloxylpentadecanoic acid.
Figure imgf000228_0003
HPLC-MS (Method C): m/z: 534 (M+23); Rt.= 6.07 min.
The intermediate aldehyde was prepared similarly as described in example 481.
Example 485 (General procedure (D)) β-fβ-t^-Dtoxothiazolidin-S-ylidenemethyOnaphthalen^-yloxylhexanoic acid.
Figure imgf000228_0004
HPLC-MS (Method C): m/z: 408 (M+23); Rt= 3.71 min. Example 486 (General procedure (D)) 4-[6-(2,4-Dioxothiazoliclin-5-ylidenemethyl)πaphthalen-2-ytoxy]butyric acid.
Figure imgf000229_0001
HPLC-MS (Method C): m/z: 380 (M+23); Rt= 3.23 min.
Example 487 (General procedure (D)) 6-[6-(2,4-Dioxothiazolidin-5-ylidenemethy[)naphthalen-2-yloxy]hexanoic acid ethyl ester.
Figure imgf000229_0002
HPLC-MS (Method C): m/z: 436 (M+23); Rt.= 4.64 min.
Example 488 (General procedure (D)) 4-[6-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-2-yloxy]butyric acid ethyl ester.
Figure imgf000229_0003
HPLC-MS (Method C): m/z: 408 (M+23); Rt= 4.28 min.
Example 489 (General procedure (D)) 2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentyl}malonic acid
Figure imgf000229_0004
HPLC-MS (Method C): m/z = 444 (M+1 ); Rt = 3,84 min. Example 490 (General procedure (D)
2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yIoxylpentyl}malonic acid diethyl ester
Figure imgf000230_0001
HPLC-MS (Method C): m/z = 500 (M+1); Rt = 5.18 min.
Example 491 (General procedure (D))
4-[4-(2,4,6-Trioxotetrahydropyrimidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid
Figure imgf000230_0002
HPLC-MS (Method C): m/z = 3;69 (M+1); Rt = 2,68 min.
Example 492
N-(3-Aminopropyl)-4-[4-(2,4-dioxothia2olidin-5-yl'tdenemethyl)-naphtha!en-1-yloxy]- butyramide
Figure imgf000230_0003
To a mixture of 4-[4-(2,4-dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]butyric acid (example 469, 5.9 g, 16.5 mmol) and 1-hydroxybenzotriazole (3.35 g, 24.8 mmol) in DMF (60 mL) was added 1-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (4.75 g, 24.8 mmol) and the resulting mixture was stirred at room temperature for 2 hours. /V-(3-amino- propylcarbamic acid tert-butyl ester (3.45 g, 19.8 mmol) was added and the resulting mixture was stirred at room temperature for 16 hours. The mixture was concentrated in vacuo and ethyl acetate and dichloromethane were added to the residue. The mixture was filtered, washed with water and dried in vacuo to afford 4.98 g (59%) of (3-{4-[4-{2,4-dioxothiazolidin- 5-ylidenemethyl)naphthalen-1-yloxy]butyrylamlno}propyl)carbamic acid tert-butyl ester.
HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.79 min.
(3-{4-[4-(2,4-Dioxothiazolϊdin-5-ylidenemethyl)πaphthaleπ-1-yloxy]butyrytamino}- propyl)carbamic acid tert-butyl ester (4.9 g, 9.5 rnmol) was added dichloromethane (50 mL) and trifluoroacetic acid (50 mL) and the resulting mixture was stirred at room temperature for 45 minutes. The mixture was concentrated in vacuo and co-evaporated with toluene. To the residue was added ethyl acetate (100 mL) and the mixture was filtered and dried in vacuo to afford the title compound as the trifluoroacetic acid salt.
HPLC-MS (Method C): m/z: 414 (M+1); Rt = 2,27 min.
Compounds of the invention includes:
Example 493
Figure imgf000232_0001
Example 494
Figure imgf000232_0002
Example 495
Figure imgf000232_0003
Example 496
Figure imgf000232_0004
Example 497
Figure imgf000232_0005
Example 498
Figure imgf000232_0006
Example 499
Figure imgf000233_0001
Example 500
Figure imgf000233_0002
Example 501
Figure imgf000233_0003
Example 502
Figure imgf000233_0004
Example 503
Figure imgf000233_0005
Example 504 (Prepared analogously to General Procedure (D)) 2-{5-[4-(2,4-Thiazolidindion-5-ylideπemethyl)naphthalen-1-yloxy]pentyl}malonic acid
Figure imgf000234_0001
A solution of 4-hydroxy-1-naphtaldehyde (1.0 g, 5.81 mmol), 2-(5-bromopentyl)ma!oπic acid diethyl ester (2.07 g, 6.68 mmol) and potassium carbonate (4.01 g, 29 mmol) in DMF (50 mL) was stirred at 100° C for 3 hours. The mixture was cooled and the salt was filtered off. The solvent was then removed under reduced pressure to afford 2.9 g of crude 2-l5-(4- formylnaphtalen-1-yloxy)pentyl]malonic acid diethyl ester which was used for the next reac¬ tion without further purification.
HPLC-MS (Method C): m/z: 401 (M+1); Rt = 5.16 min. 1H-NMR (DMSO-d6): S = 1.18 (t, 6 H), 1.39 (m, 2 H), 1.55 (m, 2 H), 1.87 (m, 4 H), 3.48 (t, 1 H), 4.13 (m, 4 H), 4.27 (t, 2 H), 7.17 (d, 1 H), 7.64(t, 1 H), 7.75 (t, 1 H), 8.13 (d, 1 H)1 8.29 (d, 1 H), 9.24 (d, 1 H), 10.19 (S, 1 H).
1.4 g (3.5 mmol) of crude 2-[5-(4-formylnaphtaleπ-1-yloxy)pentyl]rnalonic acid diethyl ester was treated with aqueous sodium hydroxide (1N, 8.75 mL, 8.75 mmol) and methanol (50 mL). The solution was stirred at 70° C for 5 hours and the mixture was concentrated under reduced pressure/ Hydrochloric acid (6 N) was added until pH <2. The resulting slurry was stirred uπtill it solidified. The crystals were filtered off, washed with water and then dried in vacuo to afford 1.1 g (92%) of 2-[5-(4-formylnaphtalen-1-yloxy)pentyl]malonic acid. The product was used in the next step without further purification.
HPLC-MS (Method C): m/z: 345 (M+1); Rt = 3.52 min. 1H-NMR(DMSO-d6): δ = 1,40 (m, 2 H), 1.55 (m, 2 H), 1.80 (m, 2 H), 1.90 (m. 2 H), 3.24 (t, 1 H), 4.29 (t, 2 H). 7.19 (d, 1 H). 7.64(t, 1 H), 7.75 (t, 1 H), 8.14 (d, 1 H), 8.30 (d, 1 H), 9.23 (d. 1 H), 10.18 (s, 1 H), 12.69 (s. 2 H).
To a solution of 2-[5-(4-formylnaphtalen-1-yloxy) pentyljmalonic acid (0.36 g, 1.05 mmol) in acetic acid (10 mL) was added 2,4-thiazolidindione (0.16 g,1.36 mmol) and piperidine (0.52 mL, 5.25 mmol). The solution was heated to 105 0C for 24 hours. After cooling to room tem¬ perature, the solvents were removed in vacuo. Water was added to the residue. The precipi- tate was filtered off and washed with water. Recrystalisation from acetonitrile afforded 200 mg (43%) of the title compound as a solid.
HPLC-MS (Method C): m/r 422 (M-C02+Na); Rt = 4.08 min. 1H-NMR(DMSO-dβ): 6 = 1.41 <m, 2 H). 1.55 (m, 4 H), 1.88 (m, 2 H), 2.23 (t. 1 H). 4.24 (t, 2 H), 7.61-7.74 (m, 3 H). 8.12 (d, 1 H), 8.28 (d, 1 H), 8.38 (s, 1 H), 12.00 (s, 1 H), 12.59 (S, 2 H).
The following compounds are commercially available and may be prepared according to general procedure (D): Example 505
Figure imgf000235_0001
Example 506
Figure imgf000235_0002
Example 507
Figure imgf000235_0003
Example 508
Figure imgf000236_0001
Example 509
Figure imgf000236_0002
Example 510
Figure imgf000236_0003
Example 511
Figure imgf000236_0004
The following salicylic acid derivatives do all bind to the His B10 Zn2+ site of the insulin hexamer: Example 512 Salicylic acid
Figure imgf000237_0001
Example 513
Thiosalicylic acid (or: 2-Mercaptobenzoic acid)
Figure imgf000237_0002
Example 514
2-Hydroxy-5-πitrobenzoic acid
Figure imgf000237_0003
Example 515 3-Nitrosalicyclic acid
Figure imgf000237_0004
Example 516
5,5'-Methylenedisalicylic acid
Figure imgf000237_0005
Example 517 2-Amiπo-5-trifluoromethyIbeπzoesyre
Figure imgf000238_0001
Example 518
2-Amino4-chlorobenzoic add
Figure imgf000238_0002
Example 519
2-Amino-5-methoxybenzoesyre
Figure imgf000238_0003
Example 520
Figure imgf000238_0004
Example 521
Figure imgf000238_0005
Example 522
Figure imgf000239_0001
Example 524
Figure imgf000239_0002
Example 525
Figure imgf000239_0003
Example 526 5-lodosalicy)ic acid
Figure imgf000239_0004
Example 527 5-Chforosalicylic acid
Figure imgf000240_0001
Example 528
1-Hydroxy-2-naphthoic acid
Figure imgf000240_0002
Example 529
3,5-Dihydroxy-2-naphthoic acid
Figure imgf000240_0003
Example 530
3-Hydroxy-2-naphthoic acid
Figure imgf000240_0004
Example 531
3,7-Dihydroxy-2-naphthoic acid
Figure imgf000240_0005
Example 532 2-Hydroxybeπzo[a]carbazα!e-3-carboxyl(c acid
Figure imgf000241_0001
Example 533
7-Bromc-3-hydroxy-2-naphthoic acid
Figure imgf000241_0002
This compound was prepared according to Murphy et a/., J. Med. Chem. 1990, 33, 171-8. HPLC-MS (Method A): m/z: 267 (M+1); Rt: = 3.78 min.
Example 534 1 ,6-Dibramo-2-hydroxynaphthalene-3-carboxylic acid
Figure imgf000241_0003
This compound was prepared according to Murphy etal., J. Med. Chem. 1990, 33, 171-8. HPLC-MS (Method A): m/z: 346 (M+1); Rt: = 4,19 min.
Example 535
7-Forrnyl-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000241_0004
A solution of 7-brαmo-3-hydroxynaphthaleπe-2-carboxylic acid (15.0 g, 56.2 mmol) (example 533) in tetrahydrofuran (100 mL) was added to a solution of lithium hydride (893 mg, 112 mmol) in tetrahydrofuran (350 mL). After 30 minutes stirring at room temperature, the result¬ ing solution was heated to 500C for 2 minutes and then allowed to cool to ambient tempera¬ ture over a period of 30 minutes. The mixture was cooled to -780C, and butyllithium (1.6 M in hexanes, 53 mL, 85 mmol) was added over a period of 15 minutes. Λ/,A/-Dimethylformamide (8.7 mL, 8.2 g, 112 mmol) was added after 90 minutes additional stirring. The cooling was discontinued, and the reaction mixture was stirred at room temperature for 17 hours before it was poured into 1 N hydrochloric acid (aq.) (750 mL). The organic solvents were evaporated in vacuo, and the resulting precipitate was filtered off and rinsed with water (3 x 100 mL) to yield the crude product (16.2 g). Purification on silica gel (dichloromethane / methanol / ace- tic acid = 90:9; 1 ) furnished the title compound as a solid.
1H-NMR (DMSO-cfe): 511.95 (1H, bs), 10.02 (1H, s), 8.61 (1H1 s), 8.54 (1H, s). 7.80 (2H1 bs), 7.24 (1H, s); HPLC-MS (Method (A)): rrr/z: 217 (M+1); Rt = 2.49 miπ.
Example 536
3-Hydroxy-7-methoxy-2-naphthoic acid
Figure imgf000242_0001
Example 537 4-Amino-2-hydroxybenzoic acid
Figure imgf000242_0002
Example 538 5-Acetylamino-2-hydroxybenzoic acid
Figure imgf000242_0003
Example 539
2-Hydroxy-5-methoxybenzoic acid
Figure imgf000242_0004
The following compounds were prepared as described below: Example 540
4-Bromo-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000243_0001
S-Hydroxynaphthalene^-carboxylic acid (3.0 g, 15.9 mmol) was suspended in acetic acid (40 mL) and with vigorous stirring a solution of bromine (817 μL, 15.9 mmol) in acetic acid (10 mL) was added drop wise during 30 minutes. The suspension was stirred at room tem¬ perature for 1 hour, filtered and washed with water. Drying in vacuo afforded 3.74 g (88%) of 4-bromo-3-hydroxynaphtha(ene-2-carboxylic acid as a solid. 1H-NMR (DMSO-dβ): δ 7.49 (1H1 1), 7.75 (1H1 t)t 8.07 (2H, T), 8.64 (1H, s). The substitution pattern was confirmed by a COSY experiment, showing connectivities between the 3 (4 hy¬ drogen) "triplets". HPLC-MS (Method A): m/z: 267 (M+1); Rt = 3.73 min.
Example 541 3-Hydroxy-4-iodonaphthalene-2-carboxylic acid
Figure imgf000243_0002
S-Hydroxynaphthalene^-carboxylic acid (0.5 g, 2.7 mmol) was suspended in acetic acid (5 mL) and with stirring iodine monochloride (135 μL, 2.7 mml) was added. The suspension was stirred at room temperature for 1 hour, filtered and washed with water. Drying afforded 0.72 g (85%) of 4-iodo-3-hydroxynaphthalene-2-carboxylic acid as a solid.
1H-NMR (DMSO-Cf6): δ 7.47 (1H, t), 7.73 (1H1 1), 7.98 (1H1 d), 8.05 (1H, d), 8.66 (1H. s). HPLC-MS (Method A): m/z: 315 (M+1); Rt = 3.94 min.
Example 542 2-Hydroxy-5-[(4-methoxyphenylarnino)rnethyl]benzoic acid
Figure imgf000244_0001
p-Anisidiπe (1.3 g, 10.6 mmol) was dissolved in methanol (20 ml.) and 5-fomnylsalicylic acid (1.75 g, 10.6 mmol)was added and the resulting mixture was stirred at room temperature for 16 hours. The solid formed was isolated by filtration, re-dissolved in N-methyl pyrrolidone (20 ml.) and methanol (2 mL). To the mixture was added sodium cyanoborohydride (1.2 g) and the mixture was heated to 700C for 3 hours. To the cooled mixture was added ethyl acetate (100 mL) and the mixture was extracted with water (100 mL) and saturated aqueous ammo¬ nium chloride (100 mL). The combined aqueous phases were concentrated in vacuo and a 2 g aliquot was purified by SepPac chromatography eluting with mixtures of aetonitrile and wa¬ ter containing 0.1% trifluoroacetic acid to afford the title compound.
HPLC-MS (Method A): m/z: 274 (M+1); Rt = 1.77 min.
1H-NMR (methanoi-cf,): δ 3.82 (3H, s). 4.45 (2H1 s), 6.96 (1H, d). 7.03 (2H1 d), 7.23 (2H1 d), 7.45 (1H1 dd), 7.92 (1 H, d).
Example 543
2-Hydroxy-5-(4-methoxyphenylsulfamoyl)benzoic acid
Figure imgf000244_0002
A solution of δ-chlrosulfonylsalicylic acid (0.96 g, 4.1 mmol) in dichloromethane (20 mL) and triethylamine (1.69 mL, 12.2 mmol) was added p-anisidine (0.49 g, 4.1 mmol) and the result¬ ing mixture was stirred at room temperature for 16 hours. The mixture was added dichloro¬ methane (50 mL) and was washed with water (2 x 100 mL). Drying (MgSO4) of the organic phase and concentration in vacuo afforded 0.57 g crude product. Purification by column chromatography on silica gel eluting first with ethyl acetate:heptane (1:1) then with methanol afforded 0.1 g of the title compound.
HPLC-MS (Method A): m/z: 346 (M+23); Rt = 2.89 min. 1H-NMR (DMSO-Cf6): δ 3.67 (3H, s), 6.62 (1H, d), 6.77 {2H, d), 6.96 (2H, d), 7.40 (1H, dd), &05 (1H. d), 9.6 (1H, bs).
General procedure (E) for preparation of compounds of general formula I4:
Figure imgf000245_0001
wherein Lea is a leaving group such as Cl, Br, I or OSO2CF3, R is hydrogen or Ci-Cβ-alkyl, optionally the two R-groups may together form a 5-8 membered ring, a cyclic boronic acid ester, and J is as defined above.
An analogous chemical transformation has previously been described in the literature (Bumagiπ et al., Tetrahedron, 1997, 53, 14437-14450). The reaction is generally known as the Suzuki coupling reaction and is generally performed by reacting an aryl halide or triflate with an arylboronic acid or a heteroarylboronic acid in the presence of a palladium catalyst and a base such as sodium acetate, sodium carbonate or sodium hydroxide. The solvent can be water, acetone, DMF, NMP, HMPA, methanol, ethanol toluene or a mixture of two or more of these solvents. The reaction is performed at room temperature or at elevated temperature.
The general procedure (E) is further illustrated in the following example: Example 544 (General Procedure (E)) 7-(4-Acetytphenyl)-3-hydraxynaphthalene-2-carboxy!ic Acid
Figure imgf000245_0002
To 7-bromc-3-hydroxynaphthalene-2-carboxylic acid (100 mg, 0.37 mmol) (example 533) was added a solution of 4-acetylphenylboronic acid (92 mg, 0.56 mmol) in acetone (2.2 mL) followed by a solution of sodium carbonate (198 mg, 1.87 mmol) in water (3.3 mL). A sus¬ pension of palladium(ll) acetate (4 mg, 0.02 mmol) in acetone (0.5 mL) was filtered and added to the above solution. The mixture was purged with N2 and stirred vigorously for 24 hours at room temperature. The reaction mixture was poured into 1 N hydrochloric acid (aq.) (60 mL) and the precipitate was filtered off and rinsed with water (3 x 40 ml_). The crude product was dissolved in acetone (25 mL) and dried with magnesium sulfate (1 h). Filtration followed by concentration furnished the title compound as a solid (92 mg). 1H-NMR (DMSO-de): £12.60 (1H, bs), 8.64 (1H, s), 8.42 (1H1 s), 8.08 (2H, d), 7.97 (2Ht d), 7.92 (2H, m), 7.33 (1H, s), 2.63 (3H, s); HPLC-MS (Method (A): mfz: 307 (M+1); Rt = 3.84 min.
The compounds in the following examples were prepared in a similar fashion. Optionally, the compounds can be further purified by recrystallization from e.g. ethanol or by chromatogra¬ phy.
Example 545 (General Procedure (E)) 3-Hydroxy-7-(3-methoxyphenyl)naphthalene-2-carboxylic acid
Figure imgf000246_0001
HPLC-MS (Method (A)): m/z: 295 (M+1); Rt = 4.60 min.
Example 546 (Generaf Procedure (E)) S-Hydroxy^-phenylnaphthalene^-carboxylic acid
Figure imgf000246_0002
HPLC-MS (Method (A)): m/z: 265 (M+1); Rt = 4.6 min.
Example 547 (General Procedure (E)) 3-Hydroxy-7-p-toIylnaphthatene-2-carboxylic acid
Figure imgf000246_0003
HPLC-MS (Method (A)): m/z: 279 (M+1); Rt = 4.95 min. Example 548 (General Procedure (E)) 7-(4-Formylphenyl)-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000247_0001
HPLC-MS (Method (A)): m/z: 293 (M+1); Rt = 4.4 min.
Example 549 (General Procedure (E)) 6-Hydroxy-[1 ,2]binaphthalenyl-7-carboxylic acid
Figure imgf000247_0002
HPLC-MS (Method (A)): m/z: 315 (M+1); Rt = 5.17 min.
Exampte 550 (General Procedure (E)) 7-(4-Carboxy-phenyl)-3-hydroxynaphthalene-2-carboxylicacid
Figure imgf000247_0003
HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 3.60 min.
Example 551 (General Procedure (E)) 7-Benzofuran-2-yl-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000247_0004
HPLC-MS (Method (A)): m/z: 305 (M+1); Rt = 4.97 min.
Example 552 (General Procedure (E)) 3-Hydroxy-7-(4-methoxyphenyl)-naphthalene-2-carboxylic acid
Figure imgf000248_0001
HPLC-MS (Method (A)): m/z: 295 (M+1); Rt = 4.68 min.
Example 553 (General Procedure (E)) 7-(3-Ethoxyphenyl)-3-hydroxynaphtha!ene-2-carboxylic acid
Figure imgf000248_0002
HPLC-MS (Method (A)): m/z: 309 (M+1); Rt - 4.89 min.
Example 554 (General Procedure (E)) 7-Beπzo[1 ,3]dioxol-5-yl-3-hydroxynaphthalene-2-carl3oxyIic acid
Figure imgf000248_0003
HPLC-MS (Method (A)): m/z: 309 (M+1); Rt = 5.61 min.
Example 555 (General Procedure (E)) 7-Biphenyl-3-yl-3-hydroxynaphthalene-2-carboxylic acid
Figure imgf000248_0004
HPLC-MS (Method (A)): m/z: 341 (M+1); Rt = 5.45 min.
General procedure (F) for preparation of compounds of general formula Is".
Figure imgf000248_0005
wherein R30 is hydrogen or d-Cβ-alkyl and T is as defined above
This general procedure (F) is further illustrated in the following example:
Example 556 (General procedure (F)) 3-Hydroxy-7-[(4-(2-propyl)pheπylamino)methyl]naphthalene-2-carboxylic Acid
Figure imgf000249_0001
7-FormyI-3-hydroxynaphthalene-2-carboxylic acid (40 mg, 0.19 mmol) (example 535) was suspended in methanol (300 μl_). Acetic acid (16 μL, 17 mg, 0.28 mmol) and 4-(2- propyljaniline (40 μL, 40 mg, 0.30 mmol) were added consecutively, and the resulting mix¬ ture was stirred vigorously at room temperature for 2 hours. Sodium cyanoborohydride (1.0 M in tetrahydrofuran, 300 μL, 0.3 mmol) was added, and the stirring was continued for an¬ other 17 hours. The reaction mixture was poured into 6 N hydrochloric acid (aq.) (6 mL), and the precipitate was filtered off and rinsed with water (3 x 2 mL) to yield the title compound (40 mg) as its hydrochloride salt. No further purification was necessary.
1H-NMR (DMSO-dβ): <510.95 (1H, bs), 8.45 (1H, s). 7.96 (1H, s), 7.78 (1H, d). 7.62 (1H. d). 7.32 (1H, s), 7.13 (2H, bd), 6.98 (2H. bd), 4.48 (2H, s), 2.79 (1H, sept), 1.14 (6H, d); HPLC- MS (Method (A)): m/z: 336 (M+1 ); Rt = 3.92 min.
The compounds in the following examples were made using this general procedure (F).
Example 557 (General procedure (F)) 7-{[(4-Bromophenyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Actd
Figure imgf000249_0002
HPLC-MS (Method C): m/z: 372 (M+1); Rt = 4.31 min.
Example 558 (General procedure (F)) 7-{[(3,5-Dichlorophenyl)amino]methyl}-3-hydroxynaphthaleπe-2-carboxylic Acid
Figure imgf000250_0001
HPLC-MS (Method C): m/z: 362 (M+1); Rt = 4.75 min.
Example 559 (General procedure (F)) T-tKBenzoVhiazol-e-yOaminolmethyiy-a-hydroxynaphthalene-Z-carboxylic Acid
Figure imgf000250_0002
HPLC-MS (Method C): m/z: 351 (M+1); Rt = 3.43 min.
Example 560 (General procedure (F)) 3-Hydrøxy-7-{Kquinoliπ-6-yl)amino]methyl}naphthaJene-2-carboxylic Acid
Figure imgf000250_0003
HPLC-MS (Method C): m/z: 345 (M+1); Rt = 2.26 min.
Example 561 (General procedure (F)) 3-Hydroxy-7-{[(4-methoxyphenyl)amino]methyl}naprithalene-2-carboxylic Acid
Figure imgf000250_0004
HPLC-MS (Method C): m/z: 324 (M+1); Rt = 2.57min.
Example 562 (General procedure (F)) 7-{[(2,3-Dihydrobenzofuran-5-ylmethyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000250_0005
HPLC-MS (Method C): m/z: 350 (M+1); Rt = 2.22 min. Example 563 (General procedure (F)) 7-{[(4-Chlorobenzyl)amino]methyl}-3-hydroxynaphthaleπe-2-cartιoxyIicAcid
Figure imgf000251_0001
HPLC-MS (Method C): m/z: 342 (M+1 ); Rt = 2.45 min.
Example 564 (General procedure (F)) 3-Hydroxy-7-{[(naphthalen-1-ylmethyl)amino]methyl}naphthalene-2-carboxylic Acid
Figure imgf000251_0002
HPLC-MS (Method C): m/z: 357 (M+1 ); Rt = 2.63 min.
Example 565 (General procedure (F)) 7-{[(Biphenyl-2-ylmethyl)amino]methyl}-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000251_0003
HPLC-MS (Method C): m/z: 384 (M+1 ); Rt = 2.90 min.
Example 566 (General procedure (F)) S-Hydroxy^-^-phenoxybenzylJaminolmethyllnaphthalene^-carboxylic Acid
Figure imgf000251_0004
HPLC-MS (Method C): m/z: 400 (M+1 ); Rt = 3.15 min.
Example 567 (General procedure (F)) 3-Hydroxy-7-{[(4-methoxybenzyl)amino]methyl}naphthalene-2-carboxylic Acid
Figure imgf000252_0001
HPLC-MS (Method C): m/z: 338 (M+1); Rt o2.32 min.
General procedure (G) for preparation of compounds of general formula I8:
Figure imgf000252_0002
wherein J is as defined above and the moiety (C-Cβ-afkanoylJijO is an anhydride.
The general procedure (G) is illustrated by the following example: Example 568 (General procedure (G)) Λ/-Acetyl-3-hydroxy-7-j;(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic Acid
Figure imgf000252_0003
3-Hydroxy-7-[(4-(2-propyl)phenylamino)methyl]naphthalene-2-carboxylic acid (25 mg, 0.07 mmol) (example 556) was suspended in tetrahydrofuran (200 μL). A solution of sodium hy- drogencarbonate (23 mg, 0.27 mmol) in water (200 μL) was added followed by acetic anhy¬ dride (14 μL, 15 mg. 0.15 mmol). The reaction mixture was stirred vigorously for 65 hours at room temperature before 6 N hydrochloric acid (4 mL) was added. The precipitate was fil¬ tered off and rinsed with water (3 x 1 mL) to yield the title compound (21 mg). No further puri- fication was necessary.
1H-NMR (DMSO-tf6): £ 10.96 (1H, bs), 8.48 (1H, s), 7.73 (1H, S)1 7.72 (1H, d), 7.41 (1H, dd), 7.28 (1H, s), 7.23 (2H, d), 7.18 (2H, d), 4.96 (2H. s). 2.85 (1H, sept). 1.86 (3H, s), 1.15 (6H, d); HPLC-MS (Method (A)): m/z: 378 (M+1); Rt = 3.90 min.
The compounds in the following examples were prepared in a similar fashion. Example 569 (General procedure (G)) Λ/-Acetyl-7-{[(4-bromophenyl)amino]methyl}-3-hydroxynaprithaleπe-2-carboxylicAcid
Figure imgf000253_0001
5 HPLC-MS (Method C): m/z: 414 (M+1 ); Rt = 3.76 min.
Example 570 (General procedure (G))
W-Acetyl-7-{[(2,3-dihydroben2ofuran-5-ylmethyl)amino]methyl}-3-hydroxynaphthalene-2- carboxylic Acid
Figure imgf000253_0002
HPLC-MS (Method C): m/z: 392 (M+1); Rt = 3.26 min.
Example 571 (General procedure (G)) N-Acetyl-7-{[(4-chlorobenzyl)amino]methyl)-3-hydroxynaphthalene-2-carboxylic Acid
Figure imgf000253_0003
HPLC-MS (Method C): m/z: 384 (M+1); Rt = 3.67 min.
Compounds of the invention may also include tetrazoles: Example 572 0 5-(3-(Naphthalen-2-yloxymethyl)-phenyl)-1H-tetrazole
Figure imgf000253_0004
To a mixture of 2-naphthol (10 g, 0.07 rnol) and potassium carbonate (10 g, 0.073 mol) in acetone (150 ππL), alpha-bromo-rn-tolunitril (13.6 g, 0.07 mol) was added in portions. The reaction mixture was stirred at reflux temperature for 2.5 hours. The cooled reaction mixture 5 was filtered and evaporated in vacuo affording an oily residue (19 g) which was dissolved in diethyl ether (150 mL) and stirred with a mixture of active carbon and MgSO4 for 16 hours. The mixture was filtered and evaporated in vacuo affording crude 18.0 g (100 %) of 3-
(naphthalen-2-yloxymethyl)-benzonitrile as a solid.
12 g of the above benzonitrile was recrystatlised from ethanol (150 mL) affording 8.3 g (69
%) of 3-(naphthalen-2-yloxymethyl)-benzonitrile as a solid.
M.p. 60 - 61 0C.
Calculated for C18H13NO:
C, 83.37 %; H, 5.05 %; N, 5.40 %; Found
C, 83.51 %; H, 5.03 %; N. 5.38 %.
To a mixture of sodium azide (1.46 g, 22.5 mmol) and ammonium chloride (1.28 g, 24.0 mmol) in dry dimethylformamide (20 mL) under an atmosphere of nitrogen, 3-(naphthalen-2- yloxymethyl)-benzonitrile (3.9 g, 15 mmol) was added and the reaction mixture was stirred at
1250C for 4 hours. The cooled reaction mixture was poured on to ice water (300 mL) and acidified to pH = 1 with 1 N hydrochloric acid. The precipitate was filtered off and washed with water, dried at 1000C for 4 hours affording 4.2 g (93 %) of the title compound.
M.p. 200 - 2020C. Calculated for Ci8H14N4O: C, 71.51 %; H, 4.67 %; N. 18.54 %; Found C, 72,11 %; H, 4.65 %; N, 17.43 %.
1H NMR (400 MHz, DMSO-dβ) δH 5.36 (s, 2H), 7.29 (dd, 1H). 7.36 (dt, 1H). 7.47 (m. 2H)1 7.66 (t. 1H), 7.74 (d, 1H), 7.84 (m, 3H), 8.02 (d, 1H), 8.22 (S1 1H).
Example 573 N-(3-(Tetrazol-5-yl)phenyl)-2-naphtoic acid amide
Figure imgf000254_0001
2-Naphtoic acid (10 g, 58 mmol) was dissolved in dichloromethane (100 mL) and N1N- dimethylformamide (0.2 mL) was added followed by thionyl chloride (5.1 ml, 70 mmol). The mixture was heated at reflux temperature for 2 hours. After cooling to room temperature, the mixture was added dropwise to a mixture of 3-aminobeπzonitril (6,90 g, 58 mmol) and triethyl amine (10 mL) in dichloromethane (75 ml_). The resulting mixture was stirred at room tem¬ perature for 30 minutes. Water (50 mL) was added and the volatiles was exaporated in_ vacuo. The resulting mixture was filtered and the filter cake was washed with water followed by heptane (2 x 25 mL). Drying in vacuo at 50 0C for 16 hours afforded 15.0 g {95 %) of N-(3- cyanophenyl)-2-naphtoic acid amide.
M.p. 138-14O 0C
The above naphthoic acid amide (10 g, 37 mmol) was dissolved in N,N-dimethylformamide (200 mL) and sodium azide (2.63 g, 40 mmol) and ammonium chloride (2.16 g, 40 mmol) were added and the mixture heated at 125 0C for 6 hours. Sodium azide (1.2 g) and ammo¬ nium chloride (0.98 g) were added and the mixture heated at 125 0C for 16 hours. After cool¬ ing, the mixture was poured into water (1.51) and stirred at room temperature for 30 minutes. The solid formed was filtered off. washed with water and dried in vacuo at 50 0C for 3 days affording 9.69 g (84 %) of the title compound as a solid which could be further purified by treatment with ethanol at reflux temperature.
1H NMR (200 MHz1 DMSO-d6): SH 7.58-7.70 (m. 3H). 7.77 (d, 1H), 8.04-8.13 (m, 5H). 8.65 (d, 1H), 10.7 (S1 1H).
Calculated for C18H13N6O, 0.75 H2O:
C, 65.74 %; H, 4.44 %; N, 21.30 %. Found:
C, 65.58 %; H, 4.50 %; N, 21.05 %.
Example 574
5-[3-(Biphenyl-4-yloxymethyl)phenyl]-1H-tetrazole
Figure imgf000255_0001
To a solution of 4-phenylphenol (10.0 g, 59 mmol) in dry N,N-dimethyl-formamide (45 mL) kept under an atmosphere of nitrogen, sodium hydride (2.82 g, 71 mmol, 60 % dispersion in oil) was added in portions and the reaction mixture was stirred until gas evolution ceased. A solution of m-cyanobenzyl bromide (13 g. 65 mmol) in dry N.N-dimethylformarnide (45 mL) was added dropwise and the reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was poured on to ice water (150 mL). The precipitate was filtered of and washed with 50 % ethanol
(3 x 50 mL), ethanol (2 x 50 mL), diethyl ether (80 mL), and dried m vacuo at 50 0C for 18 hours affording crude 17.39 g of 3-(biphenyl-4-yloxymethyl)-beπzonitrile as a solid.
1H NMR (200 MHz1 CDCI3) δH 5.14 (s, 2H)1 7.05 (m, 2H), 7.30 - 7.78 (m, 11H).
To a mixture of sodium azidβ (2.96 g, 45.6 mmol) and ammonium chloride (2.44 g, 45.6 mmol) in dry N,N-dimethylformamide (100 mL) under an atmosphere of nitrogen, 3-(biphenyl- 4-yloxyrnethyl)-benzonitrile (10.0 g, 35.0 mmol) was added and the reaction mixture was stirred at 1250C for 18 hours. The cooled reaction mixture was poured on to a mixture of 1N hydrochloric acid (60 mL) and ice water (500 mL). The precipitate was filtered off and washed with water (3 x 100 mL). 50 % ethanol (3 x 100 mL), ethanol (50 mL), diethyl ether (50 mL), ethanol (80 mL), and dried jn vacuo at 50 0C for 18 hours affording 8.02 g {70 %) of the trtle compound.
1H NMR (200 MHz, DMSO-dβ) δH 5.31 (s, 2H), 7.19 (m, 2H), 7.34 (m, 1H), 7.47 (m, 2H), 7.69 (m, 6H), 8.05 (dt, 1H), 8.24 (s, 1H).
Example 575
5-(3-Phenoxymethyl)-phenyl)-tetrazole
Figure imgf000256_0001
3-Bromomethylbenzonitrile (5.00 g, 25.5 mmol) was dissolved in N,N-dimethylfαrrnamide (50 mL), phenol (2.40 g, 25.5 mmol) and potassium carbonate (10.6 g, 77 mmol) were added. The mixture was stirred at room temperature for 16 hours. The mixture was poured into wa¬ ter (400 mL) and extracted with ethyl acetate (2 x 200 mL). The combined organic extracts were washed with water (2 x 100 mL), dried (MgSO4) and evaporated in vacuo to afford 5.19 g (97 %) 3-(phenoxymethyl)benzonitrile as an oil.
TLC: Rf = 0.38 (Ethyl acetate/heptane = 1:4) The above benzonitrile (5.19 g. 24.8 mmol) was dissolved in N.N-dimethylformamide (100 mL) and sodium azide (1.93 g, 30 mmol) and ammonium chloride (1.59 g, 30 mmol) were added and the mixture was heated at 140 0C for 16 hours. After cooling, the mixture was poured into water (800 mL). The aqeous mixture was washed with ethyl acetate (200 mL). The pH of the aqueous phase was adjusted to 1 with 5 N hydrochloric acid and stirred at room temperature for 30 minutes. Filtration, washing with water and drying in vacuo at 500C afforded 2.06 g (33 %) of the title compound as a solid.
1H NMR (200 MHz, CDCI3 + DMSO-dθ) δH 5.05 (ε, 2H), 6.88 (m, 3H), 7.21 (m. 2H). 7.51 (m,
2H), 7.96 (dt, 1H), 8.14 (S1 1H).
Example 576
5-[3-(Biphenyl-4-ylmethoxy)phenyl]-1H-tetrazole
Figure imgf000257_0001
To a solution of 3-cyanophenol (5.0 g, 40.72 mmol) in dry N.N-dimethylformamide (100 mL) kept under an atmosphere of nitrogen, sodium hydride (2 g, 48.86 mmol, 60 % dispersion in oil) was added in portions and the reaction mixture was stirred until gas evolution ceased, p- Phenylbenzyl chloride (9.26 g, 44.79 mmol) and potassium iodide (0.2 g, 1.21 mmol) were added and the reaction mixture was stirred at room temperature for 60 hours. The reaction mixture was poured on to a mixture of saturated sodium carbonate (100 mL) and ice water (300 mL). The precipitate was filtered of and washed with water (3 x 100 mL), n-hexane (2 x 80 mL) and dried m vacuo at 50 0C for 18 hours affording 11.34 g (98 %) of 3-(bipheny!-4- ylmethoxy)-benzonitrile as a solid.
To a mixture of sodium azide (2.37 g, 36.45 mmol) and ammonium chloride (1.95 g, 36.45 mmol) in dry N.N-dimethylforrnamide (100 mL) under an atmosphere of nitrogen, 3-(biphenyl- 4-y[methoxy)-benzonitrile (8.0 g, 28.04 mmol) was added and the reaction mixture was stirred at
1250C for 18 hours. To the cooled reaction mixture water (100 mL) was added and the reac¬ tion mixture stirred for 0.75 hour. The precipitate was filtered off and washed with water, 96 % ethanol (2 x 50 mL), and dried ]n vacuo at 500C for 18 hours affording 5.13 g (56 %) of the title compound. 1H NMR (200 MHz, DMSO-dβ) δH 5.29 <s, 2H)1 7.31 (ddr 1H), 7.37 - 7.77 (m. 12H).
Example 577 5-[4-(Biphenyt-4-ylmethoxy)-3-methoxyphenyl]-1H-tetrazol
Figure imgf000258_0001
This compound was made similarly as described in example 576.
Figure imgf000258_0002
Example 579 5-(2-Naph1ylmethylM H-tetrazole
Figure imgf000258_0003
This compound was prepared similarly as described in example 572, step 2.
Example 580
5-(1 -Naphtylmethyl)-1 H-tetrazole
Figure imgf000258_0004
This compound was prepared similarly as described in example 572, step 2.
Example 581 5-[4-(Biphenyl-4-yloxymethyl)phenyl]-1H-tetrazole
Figure imgf000259_0001
A solution of alpha-bromo-p-tolunitrile (5.00 g, 25.5 mmol), 4-phenylphenol (4.56 g, 26.8 mmol), and potassium carbonate (10.6 g, 76.5 mmol) in N,N-dimethylformamide (75 mL) was stirred vigorously for 16 hours at room temperature. Water (75 mL) was added and the mix- ture was stirred at room temperature for 1 hour. The precipitate was filtered off and washed with thoroughly with water. Drying in vacuo over night at 500C afforded 7.09 g (97 %) of 4- (biphenyl-4-yloxymethyl)benzonitrile as a solid.
The above benzonitrile (3.00 g, 10.5 mmol) was dissolved in N,N-dimethylformamide (50 mL), and sodium azide (1.03 g, 15.8 mmol) and ammonium chloride (0.84 g, 15.8 mmol) were added and the mixture was stirred 16 hours at 1250C. The mixture was cooled to room temperature and water (50 mL) was added. The suspension was stirred overnight, filtered, washed with water and dried in vacuo at 500C for 3 days to give crude 3.07 g (89 %) of the title compound. From the mother liquor crystals were colected and washed with water, dried by suction to give 0.18 g
(5 %) of the title compound as a solid.
1H NMR (200 MHz, DMSO-dθ): SH 5.21 (s. 2H), 7.12 (d, 2H), 7.30 (t, 1H).7.42 (t, 2H), 7.56- 7.63 (m, 6H), 8.03 (d, 2H).
Calculated for C20H16N4O, 2H2O:
C, 65.92 %; H, 5.53 %; N, 15.37 %. Found:
C, 65.65 %; H, 5.01 %; N, 14.92 %.
Example 582
Figure imgf000259_0002
This compound was prepared similarly as described in example 576. Example 583
Figure imgf000260_0001
Example 584
Figure imgf000260_0002
Example 585
Figure imgf000260_0003
Example 586 5-(3-(Biphenyl-4-yloxymethyl)-benzyl)-1H-tetrazole
Figure imgf000260_0004
Example 587
5-(1 -Naphthyl)-1 H-tetrazole
Figure imgf000260_0005
This compound was prepared similarly as described in example 572, step 2.
Example 588 5-[3-Methoxy-4-(4-methylsuIfonylbenzyloxy)pheπyl]-1H-tetrazole
Figure imgf000261_0001
This compound was made similarly as described in example 576.
Example 589 5-(2-Naphthyl)-1 W-tetrazole
Figure imgf000261_0002
This compound was prepared similarly as described in example 572, step 2.
Example 590 2-Amino-N-(1H-tetrazol-5-yl)-benzamide
Figure imgf000261_0003
Example 591 5-(4-Hydroxy-3-methoxyphenyl)-1H-tetrazo!e
Figure imgf000261_0004
This compound was prepared similarly as described in example 572, step 2. Example 592
4-(2H-Tetrazol-5-ylmethoxy)benzoic acid
Figure imgf000262_0001
To a mixture of methyl 4-hydroxybenzoate (30.0 g, 0.20 mol), sodium iodide (30.0 g, 0.20 mo!) and potassium carbonate (27.6 g, 0.20 mot) in acetone (2000 mL) was added chloroacetonitrile (14.9 g , 0.20 mol). The mixture was stirred at RT for 3 days. Water was added and the mixture was acidified with 1N hydrochloric acid and the mixture was extracted with diethyl ether. The combined organic layers were dried over Na2SO4 and concentrated in vacuo. The residue was dissolved in acetone and chloroacetonitrile (6.04 g,0.08 mol), so- dium iodide (12.0 g, 0.08 mol) and potassium carbonate (11.1 g, 0.06 mol) were added and the mixture was stirred for 16 hours at RT and at 60 °C. More chloroacetonitrile was added until the conversion was 97%. Water was added and the mixture was acidified with 1 N hy¬ drochloric acid and the mixture was extracted with diethyl ether. The combined organic lay¬ ers were dried over Na2SO4 and concentrated in vacuo to afford methyl 4- cyanornethyloxybenzoate in quantitative yield. This compound was used without further puri¬ fication in the following step.
A mixture of methyl 4-cyanomethyloxybenzoate (53.5 g,0.20 mol), sodium azide (16.9 g, 0.26 mol) and ammonium chloride (13.9 g, 0.26 mol) in DMF 1000 (mL) was refluxed overnight under N2. After cooling, the mixture was concentrated in vacuo. The residue was suspended in cold water and extracted with ethyl acetate. The combined organic phases were washed with brine, dried over Na2SO4 and concentrated in vacuo, to afford methyl 4-(2H-tetra∑ol-5- ylmethoxy)benzoate. This compound was used as such in the following step.
Methyl 4-(2H-Tetrazol-5-ylmethoxy)-benzoate was refluxed in 3N sodium hydroxide. The re¬ action was followed by TLC (DCMiMeOH = 9:1 ). The reaction mixture was cooled, acidified and the product filtered off. The impure product was washed with DCM1 dissolved in MeOH, filtered and purified by column chromatography on silica gel (DCM:MeOH = 9:1).The result¬ ing product was recrystallised from DCM:MeOH=95:5. This was repeated until the product was pure. This afforded 13.82 g (30 %) of the title compound.
1H-NMR (DMSO-de): 4.70 (2H. s), 7.48 (2H1 d), 7.73 (2H, d), 13 (1 H, bs). Example 593
4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoic acid
Figure imgf000263_0001
To a solution of sodium hydroxide (10.4 g, 0.26 rnol) in degassed water (600 mL) was added 4-mercaptobenzoic acid (20.0 g, 0.13 mol). This solution was stirred for 30 minutes. To a so¬ lution of potassium carbonate (9.0 g, 65 mmol) in degassed water (400 mL) was added chloroacetonitrile (9.8 g, (0.13 mol) portion-wise. These two solutions were mixed and stirred for 48 hours at RT under N2. The mixture was filtered and washed with heptane. The aque¬ ous phase was acidified with 3N hydrochloric acid and the product was filtered off, washed with water and dried, affording 4-cyanomethylsulfanylbenzoic acid (27.2 g, 88%). This com¬ pound was used without further purification in the following step.
A mixture of 4-cyanomethylsulfanyJbenzoic acid (27.2 g, 0.14 mol), sodium azide (11.8 g, 0,18 mol) and ammonium chloride (9.7 g, 0.18 mol) in DMF (1000 mL) was refluxed over- night under N2. The mixture was concentrated in vacuo. The residue was suspended in cold water and extracted with diethyl ether. The combined organic phases were washed with brine, dried over Na2SO4 and concentrated in vacuo. Water was added and the precipitate w'is filtered off. The aqueous layer was concentrated in vacuo, water was added and the precipitate filtered off. The combined impure products were purified by column chromatogra- phy using DCM:MeOH = 9:1 as eluent, affording the title compound (5.2 g, 16%).
1H-NMR (DMSO-cfe): 5.58 (2H. s), 7.15 (2H, d), 7.93 (2H, d), 12.7 (1H. bs).
Example 594 3-(2H-Tetrazol-5-yl)-9H-carbazole
Figure imgf000263_0002
3-Bromo-9H-carbazole was prepared as described by Smith ef a/, in Tetrahedron 1992, 48, 7479-7488. A solution of 3-bromo-9H-carbazole (23.08 g, 0.094 mol) and cuprous cyanide (9.33 g, 0.103 mol) in Λ/-methyl-pyrrolidone (300 ml) was heated at 2000C for 5 h. The cooled reaction mix¬ ture was poured on to water (600 ml) and the precipitate was filtered off and washed with ethyl acetate (3 x 50 ml). The filtrate was extracted with ethyl acetate (3 x 250 ml) and the combined ethyl acetate extracts were washed with water (150 ml), brine (150 ml), dried
(MgSO4) and concentrated in vacuo. The residue was crystallised from heptanes and recrys- tallised from acetonitrile (70 ml) affording 7.16 g (40 %) of 3-cyaπo-9H-carbazole as a solid. M.p. 180 - 181 0C.
3-Cyano-9H-carbazole (5.77 g, 30 mmol) was dissolved in Λ/,A/-dimethylformamide (150 ml), and sodium azide (9.85 g, 152 mmol), ammonium chloride (8.04 g, 150 mmol) and lithium chloride (1.93 g, 46 mmol) were added and the mixture was stirred for 20 h at 125 0C. To the reaction mixture was added an additional portion of sodium azide (9.85 g. 152 mmol) and ammonium chloride (8.04 g, 150 mmol) and the reaction mixture was stirred for an additional 24 h at 1250C. The cooled reaction mixture was poured on to water (500 ml). The suspen¬ sion was stirred for 0.5 h, and the precipitate was filtered off and washed with water (3 x 200 ml) and dried in vacuo at 50 0C. The dried crude product was suspended in diethyl ether (500 ml) and stirred for 2 h, filtered off and washed with diethyl ether (2 x 200 ml) and dried in vacuo at 500C affording 5.79 g (82 %) of the title compound as a solid.
1H-NMR (DMSO-ds): δ 11.78 (1 H, bs). 8.93 (1 H, d), 8.23 (1 H. d). 8.14 (1 H. dd), 7.72 (1 H1 d), 7.60 (1H1 d), 7.49 (1H1 1), 7.28 (1H, t); HPLC-MS (Method C): m/z: 236 (M+ 1); Rt = 2.77 min.
The following commercially available tetrazoles do all bind to the His B10 Zn2+ site of the insulin hexameπ
Example 595 5-(3-ToIyI)-I H-tetrazole
Figure imgf000264_0001
Example 596 5-(2-Bromophenyl)tetrazole
Figure imgf000265_0001
Example 597 5-(4-Ethoxalylamino-3-nitrophenyl)tetrazole
Figure imgf000265_0002
Example 598
Figure imgf000265_0003
Example 599
Figure imgf000265_0004
Example 600
Figure imgf000266_0001
Example 601
Figure imgf000266_0002
Example 602 Tetrazole
Figure imgf000266_0003
Example 603 5-Methyltetrazole
Figure imgf000266_0004
Example 604
5-Benzyl-2H-tetrazσle
Figure imgf000266_0005
Example 605 4-(2H-TetrazoI-5-yI)benzoic acid
Figure imgf000266_0006
Example 606 5-Phenyl-2H-tetrazo!e
Figure imgf000267_0001
Example 607
5-(4-Chlorophenylsulfany1methyl)-2H-tetrazole
Figure imgf000267_0002
Example 608
5-(3-Benzyloxyphenyl)-2H-tetrazole
Figure imgf000267_0003
Example 609 2-Phenyl-6-(1H-tetrazoi-5-yl)-chromen-4-one
Figure imgf000267_0004
Example 610
Figure imgf000267_0005
Example 611
Figure imgf000268_0001
Example 612
Figure imgf000268_0002
Example 613
Figure imgf000268_0003
Example 614
Figure imgf000268_0004
Example 615 5-{4-Bromo-phenyl)- 1 H-tetrazole
Figure imgf000269_0001
Example 616
Example 617
Figure imgf000269_0003
Example 61 S
Figure imgf000269_0004
Example 619
Figure imgf000269_0005
Example 620
Figure imgf000270_0001
Example 621
Figure imgf000270_0002
Example 622
Figure imgf000270_0003
Example 623
Figure imgf000270_0004
Example 624
Figure imgf000270_0005
Example 625
Figure imgf000271_0001
Example 626
Figure imgf000271_0002
Example 627
Figure imgf000271_0003
Example 628
Figure imgf000271_0004
Example 629
Figure imgf000271_0005
Example 630
Figure imgf000272_0001
Example 632
Figure imgf000272_0002
Example 633
Figure imgf000272_0003
Example 634
Figure imgf000273_0001
Example 635
Figure imgf000273_0002
Example 636
Figure imgf000273_0003
Example 637
Figure imgf000273_0004
Example 638
Figure imgf000274_0001
Example 639
Figure imgf000274_0002
Example 640
Figure imgf000274_0003
Example 641
Figure imgf000274_0004
Example 642
Figure imgf000275_0001
Example 643
Figure imgf000275_0002
Example 644
Figure imgf000275_0003
Example 645
Figure imgf000275_0004
Example 646 5-(2,6-Dichforobenzy!)-2W-tetrazote
Figure imgf000276_0001
General procedure (H) for preparation of compounds of genera! formula Ir:
NaCBH: 3
Figure imgf000276_0002
I7 wherein K1 M, and T are as defined above.
The reaction is generally known as a reductive alkylation reaction and is generally performed by stirring an aldehyde with an amine at low pH (by addition of an acid, such as acetic acid or formic acid) in a solvent such as THF, DMF, NMP1 methanol, ethanol, DMSO, dichloro- methane, 1 ,2-dichloroethane, trimethyl orthoformate, triethyl orthofσrmate, or a mixture of two or more of these. As reducing agent sodium cyano borohydride or sodium triacetoxy borohydride may be used. The reaction is performed between 2O0C and 12O0C, preferably at room temperature.
When the reductive alkylation is complete, the product is isolated by extraction, filtration, chro¬ matography or other methods known to those skilled in the art.
The general procedure (H) is further illustrated in the following example 647:
Example 647 (General procedure (H))
Biphenyl-4-ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amlne
Figure imgf000276_0003
A solution of 5-(3-aminophenyl)-2H-tetrazole (example 874, 48 mg, 0.3 mmol) in DMF (250 μl_) was mixed with a solution of 4-biphenylylcarbaldehyde (54 mg, 0.3 mmol) in DMF (250 μL) and acetic acid glacial (250 μL) was added to the mixture followed by a solution of so- dium cyano borohydride (15 mg, 0.24 mmol) in methanol (250 μL). The resulting mixture was shaken at room temperature for 2 hours. Water (2 mL) was added to the mixture and the re¬ sulting mixture was shaken at room temperature for 16 hours. The mixture was centrifugated (6000 rpm, 10 minutes) and the supernatant was removed by a pipette. The residue was washed with water (3 mL), centrifugated (6000 rpin, 10 minutes) and the supernatant was removed by a pipette. The residue was dried in vacua at 400C for 16 hours to afford the title compound as a solid.
HPLC-MS (Method C): m/z: 328 (M+1), 350 (M+23); Rt = 4.09 min.
Example 648 (General procedure (H)) Benzyl-[3-(2H-tetrazol-5-y!)phenyl]amine
Figure imgf000277_0001
HPLC-MS (Method D): m/z: 252 (M+1); Rt = 3,74 min.
Example 649 (General procedure (H)) (4-Methoxybenzyl)-[3-(2W-tetrazol-5-yl)phenyl]amiπe
Figure imgf000277_0002
HPLC-MS (Method D): m/z: 282,2 (M+1); Rt = 3.57 min.
Example 650 (General procedure (H)) 4-{[3-(2W-Tetrazok5-yl)phenylamino]methyl}phenol
Figure imgf000277_0003
HPLC-MS (Method D): m/z: 268,4 (M+1); Rt = 2,64 min.
Example 651 (General procedure (H)) (4-Nitrobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000278_0001
HPLC-MS (Method D): m/z: 297,4 (M+ 1); Rt = 3,94 min.
Example 652 (General procedure (H)) (4-Chlorobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000278_0002
HPLC-MS (Method D): m/z: 287,2 (M+1); Rt = 4,30 min.
Example 653 (General procedure (H)) (2-Chlorobenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000278_0003
HPLC-MS (Method D): m/z: 286 (M+1); Rt = 4,40 min.
Example 654 (General procedure (H)) (4-Bromobenzy[)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000278_0004
HPLC-MS (Method D): mfø:332 (M+1); Rt = 4,50 min.
Example 655 (General procedure (H)) (3-Benzyloxybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000278_0005
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,94 min. Example 656 (General procedure (H)) Naphthalen-1-ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000279_0001
HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min.
Example 657 (General procedure (H)) Naphthalen-2-ylmethyl-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000279_0002
HPLC-MS (Method D): mfe 302 (M+1); Rt = 4.60 min.
Example 658 (General procedure (H)) 4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}benzoic acid
Figure imgf000279_0003
HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3,24 min.
Example 659 (General procedure (H)) [3-(2H-Tetrazol-5-yl)-phenyl]-[3-(3-trifluoromethyl-phenoxy)benzyl]amine
<VΛ ,F
HPLC-MS (Method D): m/z: 4-12 (M+1); Rt = 5,54 min.
Example 660 (General procedure (H)) (3-Phenoχybenzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000279_0004
HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,04 min.
Example 661 (General procedure (H)) (4-Phenoxy-benzyl)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000280_0001
HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,00 min.
Example 662 (General procedure (H)) (4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl}phenoxy)acetic acid
Figure imgf000280_0002
HPLC-MS (Method D): m/z: 326 (M+1); Rt = 3,10 min.
Example 663 (General procedure (H)) (4-Benzyloxybenzy!)-[3-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000280_0003
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,97 min.
Example 664 (General procedure (H)) 3-(4-{[3-(2H-Tetrazol-5-yl)phenylamino]methyl)phenyl)acrylic acid
Figure imgf000280_0004
HPLC-MS (Method D): m/z: 322 (M+1); Rt = 3,60 min.
Example 665 (General procedure (H)) Dimethyl-(4-{[3-(2H-tetrazol-5-yl)phenylamino]methyl}naphthalen-1-yl)amine
Figure imgf000281_0001
HPLC-MS (Method D): m/z: 345 (M+1); Rt = 3,07 min.
Example 666 (General procedure (H)) (4'-MethoxybiphenyI-4-ylmethyl)-[3-(2H-tetrazo!-5-yl)prιenyI]amine
Figure imgf000281_0002
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4,97 min.
Example 667 (General procedure (H)) (2'-ChIorobiphenyl-4-ylmethylH3-(2W-tetrazol-5-yl)pheπyl]amine
Figure imgf000281_0003
HPLC-MS (Method D): m/z: 362 (M+1 ); Rt = 5.27 min.
Example 668 (General procedure (H)) Benzyl-[4-(2W-tetrazol-5-yl)pheπyl]amine
Figure imgf000281_0004
For preparation of starting material, see example 875. HPLC-MS (Method D): m/z: 252 (M+1); Rt - 3,97 min.
Example 669 (General procedure (H))
(4-MethoxybenzylM4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000282_0001
HPLC-MS (Method D): m/z: 282 (M+1); Rt = 3.94 min.
Example 670 (General procedure (H)) 4-{[4-(2H-Tetrazo»-5-yl)phenytøminolmethyl}phenol
Figure imgf000282_0002
HPLC-MS (Method D): m/z: 268 (M+1); Rt = 3,14 min.
Example 671 (General procedure (H)) (4-Nitrobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000282_0003
HPLC-MS (Method D): m/z: (M+1); Rt = 3,94 min.
Example 672 (General procedure (H)) (4-Chlorαbenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000282_0004
HPLOMS (Method D): m/z: (M+1); Rt = 4,47 min.
Example 673 (General procedure (H)) (2-Chlorobenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000282_0005
HPLC-MS (Method D): m/z: 286 (M+1); Rt = 4,37 min.
Example 674 (General procedure (H)) (4-BromobenzylH4-(2H-tetrazol-5-y!)phenyl_amine
Figure imgf000283_0001
HPLC-MS (Method D): m/z: 331 (M+1); Rt = 4,57 min.
Example 675 (General procedure (H)) (3-Benzyloxybenzyl)-[4-{2H-tetraεol-5-yl)phenyl]amine
Figure imgf000283_0002
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,07min.
Example 676 (General procedure (H)) Naphthalen-1~ylmethyl-[4-(2M-tetrazo!-5-yl)phenyl]amine
Figure imgf000283_0003
HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min.
Example 677 (General procedure (H)) Naphthalen-2-ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]arnine
Figure imgf000283_0004
HPLC-MS (Method D): m/z: 302 (M+1); Rt = 4,70 min.
Example 678 (General procedure (H)) Biphenyl-4-ylmethyl-[4-(2H-tetrazol-5-yl)phenyl]amine
HPLC-MS (Method D): m/z: 328 (M+1); Rt = 5,07 min.
Example 679 (General procedure (H)) 4-{[4-(2H-Tetrazol-5-yl)phenyiamino]methyl}benzoic acid
Figure imgf000284_0002
HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3,34 min.
Example 680 (General procedure (H)) [4-(2H-Tetrazol-5-yl)phenyl]-[3-(3-trifluoromethylphenoxy)benzy1]amine
Figure imgf000284_0003
HPLC-MS (Method D): m/z: 412 (M+1); Rt= 5,54 min.
Example 681 (General procedure (H)) (3-Phenoxybenzyl)-[4-(2H-tetrazol-5-yl)phenyl]amine
Figure imgf000284_0004
HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5.07 min.
Example 682 (General procedure (H)) (4-Phenoxybenzyl)-[4-(2H-tetrazol-5-yl)-phenyl]-amine
Figure imgf000284_0005
HPLC-MS (Method D): m/z: 344 (M+1); Rt = 5,03 min. Example 683 (General procedure (H)) 3-{[4-(2H-Tetrazol-5-yl)phenylamino]rnethyl}benzoic acid
Figure imgf000285_0001
HPLC-MS (Method D): m/z: 286 (M+1); Rt = 3.47 min.
Example 684 (General procedure (H)) (4-{[4-(2H-Tetrazol-5-yl)pheπylaminolmethyl}phenoxy)acetic acid
Figure imgf000285_0002
HPLC-MS (Method D): m/z: 326 (M+1 ); Rt = 3,40 min.
Example 685 (General procedure (H)) (4-Beπ2yloxybeπzy()-[4-(2W-tetrazo[-5-yl)phenyfJarnine
Figure imgf000285_0003
HPLC-MS (Method D): m/z: 358 (M+1 ); Rt = 5,14 min.
Example 686 (General procedure (H)) 3-(4-{[4-(2W-Tetrazol-5-yl)phenylamino3methyl}phenyl)acrylic acid
Figure imgf000285_0004
HPLC-MS (Method D): m/r. 322 (M+1); Rt = 3,66 min.
Example 687 (General procedure (H)) Dimethyl-(4-{[4-(2H-tetrazol-5-yl)phenyiamino]methyl}naphthalen-1-yl)amine
Figure imgf000286_0001
HPLC-MS (Method D): m/z: 345 (M+1); Rt = 3,10 min.
Example 688 (General procedure (H)) (4'-Methoxybiphenyl-4-yImethyl)-[4-(2H-tetrazol-5-yl)phenyl]amiπe
Figure imgf000286_0002
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 5,04 min.
Example 689 (General procedure (H)) (2'-Chlorobiphenyl-4-ylmethyl)-[4-(2H-tetrazol-5-y1)-phenyl]-amine
HPLC-MS (Method D): m/z: 362 (M+1); Rt = 5,30 min.
General procedure (I) for preparation of compounds of general formula I8:
HOAt
Figure imgf000286_0004
I, wherein K, M and T are as defined above.
This procedure is very similar to general procedure (A), the only difference being the carbox- ylic acid is containing a tetrazole moiety. When the acylation is complete, the product is iso- lated by extraction, filtration, chromatography or other methods known to those skilled in the art.
The general procedure (I) is further illustrated in the following example 690: Example 690 (General procedure (I)) 4-[4-(2H-Tetrazol-5-yl)benzoylamino]benzoic acid
Figure imgf000287_0001
To a solution of 4-(2H-tetrazol-5-yl)benzoic acid (example 605, 4 mmol) and HOAt (4.2 mmol) in DMF (6 ml_) was added 1-ethyl-3-(3'-dimethylamiπopropyl)carbodiimide hydrochlo¬ ride (4.2 mmol) and the resulting mixture was stirred at room temperature for 1 hour. An a!quot of this HOAt-ester solution (0.45 mL) was mixed with 0.25 mL of a solution of 4- aminobenzoic acid (1.2 mmol in 1 mL DMF). (Anilines as hydrochlorides can also be utilised, a slight excess of triethylamine was added to the hydrochloride suspension in DMF prior to mixing with the HOAt-ester.) The resulting mixture was shaken for 3 days at room tempera¬ ture. 1N hydrochloric acid (2 mL) was added and the mixture was shaken for 16 hours at room temperature. The solid was isolated by centrifugation (alternatively by filtration or ex¬ traction) and was washed with water (3 mL). Drying in vacuo at 400C for 2 days afforded the title compound.
HPLC-MS (Method D): m/z: 310 (M+1); Rt = 2.83 min.
Example 691 (General procedure (I)) 3-[4-(2H-Tetrazol-5-yl)benzoylamino]benzoic acid
Figure imgf000287_0002
HPLC-MS (Method D): m/z: 310 (M+1); Rt = 2.89 min.
Example 692 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-y|)benzoylamino]phenyl}acrylic acid
Figure imgf000287_0003
HPLC-MS (Method D): m/z: 336 (M+1); Rt = 3.10 min. Example 693 (General procedure (I)) 3.{4~[4.(2H-TetrazQl-5-yl)beπzoylaminolphenyl}propionic acid
Figure imgf000288_0001
HPLC-MS (Method D): m/z: 338 (M+1 ); Rt = 2.97 min.
Example 694 (General procedure (I)) 3-Methoxy-4-[4-(2W-tetrazol-5-yl)benzoylamiπo]benzoic acid
Figure imgf000288_0002
HPLC-MS (Method D): m/z: 340 (M+ 1 ); Rt - 3.03 min.
Example 695 (General procedure (I)) N-(4-Benzyloxyphenyl)-4-(2W-tetrazot-5-yl)benzamide
Figure imgf000288_0003
HPLC-MS (Method D): m/z: 372 (M+1 ); Rt = 4.47 min.
Example 696 (General procedure (I)) Λ/-(4-PhenoxyphenylH-(2H-tetrazol-5-yl)benzamide
Figure imgf000288_0004
HPLC-MS (Method D): m/z: 358 (M+1); Rt = 4.50 min.
Example 697 (General procedure (I)) N-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-yl)benzamide
Figure imgf000289_0001
HPLC-MS {Method D): m/z: 354 (M+1); Rt = 4.60 min.
Example 698 (General procedure (I)) Λ/-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-yl)benzamide
Figure imgf000289_0002
HPLC-MS (Method D): m/z: 383 (M+1); Rt = 4.60 min.
Example 699 (General procedure (I)) Λ/-Phenyl-4-(2tf-tetrazol-5-yl)benzamide
Figure imgf000289_0003
HPLC-MS (Method D): m/z: 266 (M+1); Rt = 3.23 min.
Example 700 (General procedure (I)) 4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]benzoic acid
Figure imgf000289_0004
The starting material was prepared as described in example 592. HPLC-MS (Method D): m/z: 340 (M+1); Rt = 2.83 min.
Example 701 (General procedure (I))
3-[4-(2H-Tetrazol-5-yfrnethoxy)beπzoylamiπo]benzoic acid
Figure imgf000290_0001
HPLC-MS (Method D): m/z: 340 (M+1); Rt = 2.90 min.
Example 702 (General procedure (I)) s^-^^aH-Tetrazol-S-ylmethoxyJbenzoylaminolphenylJacrylic acid
Figure imgf000290_0002
HPLC-MS (Method D): m/z: 366 (M+1); Rt = 3.07 min.
Example 703 (General procedure (I)) 3-{4-[4-(2H-Tetrazol-5-ylmethoxy)benzoylamino]phenyl}propionic acid
Figure imgf000290_0003
HPLC-MS (Method D): m/z: 368 (M+1); Rt = 2.97 min.
Example 704 (General procedure (I)) 3-Methoxy-4-[4-(2H-tetrazol-5-ylmethoxy)benzoylamino]benzoic acid
Figure imgf000290_0004
HPLC-MS (Method D): m/z: 370 (M+1); Rt = 3.07 min.
Example 705 (General procedure (I)) Λ/-(4-Benzyloxyphenyl)-4-(2H-tetrazσl-5-ylmethoxy)benzamide
Figure imgf000290_0005
HPLC-MS (Method D): m/z: 402 (M+1); Rt = 4.43 min. Example 706 (General procedure (I)) W-(4-Phenoxyphenyl)-4-(2H-tetrazo1-5-ylrnethoxy)benzamide
Figure imgf000291_0001
HPLC-MS (Method D): m/z: 388 (M+1 ); Rt = 4.50 min.
Example 707 (General procedure (I)) A/-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-ylmethoxy)benzamide
Figure imgf000291_0002
HPLC-MS (Method D): m/z: 384 (M+1 ); Rt = 4.57 min.
Example 708 (General procedure (I)) W-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazol-5-ylrnethoxy)benzamide
Figure imgf000291_0003
HPLC-MS (Method D): m/z: 413 (M+1 ); Rt = 4.57 min.
Example 709 (General procedure (I)) W-Phenyl-4-(2H-tetrazol-5-ylmethoxy)benzamide
Figure imgf000291_0004
HPLC-MS (Method D): m/z: 296 (M+1); Rt = 3.23 min.
Example 710 (General procedure (I)) 4-t4-(2H-TetrazoI-5-ylmethylsulfanyl)benzoylamino]benzoic acid
Figure imgf000292_0001
The starting material was prepared as described in example 593. HPLC-MS (Method D): m/z: 356 (M+1); Rt= 2.93 min.
Example 711 (General procedure (I))
3-[4-(2H-Tetra2ol-5-y(methytsulfanyl)benzoylaminolfaenzoic add
Figure imgf000292_0002
HPLC-MS (Method D): m/r. 356 (M+1); Rt= 3.00 min.
Example 712 (General procedure (I))
3-{4-.[4.(2H-Tetrazo1-5-ylmethylsulfanyJ)ben2oylamino)phenyl}acrylic acJd
Figure imgf000292_0003
HPLC-MS (Method D): m/z: 382 (M+1); Rt = 3.26 min.
Example 713 (General procedure (I))
3-{4^4-(2H-Tetrazol-5-ylmethylsulfanyl)benzoylamino]phenyl}ρropionic acid
Figure imgf000292_0004
HPLC-MS (Method D): m/z: 364 (M+1); Rt = 3.10 min.
Example 714 (General procedure (I))
3-Methoxy-4-[4-(2H-tetrazol-5-ylmethylsulfanyl)benzoylamino]benzoic acid
Figure imgf000292_0005
HPLC-MS (Method D): m/z: 386 (M+1); Rt = 3.20 min.
Example 715 (General procedure (I)) Λ/-(4-Benzyloxyphenyl)-4-(2H-tetrazol-5-y1methylsulfanyI)benzamide
Figure imgf000293_0001
HPLC-MS (Method D): m/z: 418 (M+1); Rt = 4.57 min.
Example 716 (General procedure (I)) N-(4-Phenoxyphenyl)-4-(2H-tetrazol-5-ylmethylεulfanyl)benzamide
Figure imgf000293_0002
HPLC-MS (Method D): m/z: 404 (M+1); Rt = 4.60 min.
Example 717 (General procedure (I)) N-(9H-Fluoren-2-yl)-4-(2H-tetrazol-5-ylmethylsulfanyl)benzamide
Figure imgf000293_0003
HPLC-MS (Method D): m/z: 400 (M+1); Rt = 4.67 min.
Example 718 (General procedure (I)) Λ/-(9-Ethyl-9H-carbazol-2-yl)-4-(2H-tetrazo1-5-ylmethylsu1fanyl)benzamide
Figure imgf000293_0004
HPLC-MS (Method D): m/z: 429 (M+1); Rt = 4.67 min.
Example 719 (General procedure (I)) N-PhenyI-4-(2H-tetrazol-5-ylmβthylsulfanyl)benzarnide
Figure imgf000294_0001
HPLOMS (Method D): m/z: 312 (M+1); Rt = 3.40 min.
General procedure (J) for solution phase preparation of amides of general formula I9:
Figure imgf000294_0002
wherein T is as defined above.
This general procedure (J) is further illustrated in the following example.
Example 720 (General procedure (J)). 9-(3-Chlσrobeπzyl}-3-(2tt-tetrazol-5-yl)-9H-carbazote
Figure imgf000294_0003
3-(2H-Tetrazo1-5-yl)-9H-carbazole (example 594. 17 g, 72.26 mmol) was dissolved in N,N- dimethylformamide (150 mL). Triphenylmethyl chloride (21.153 g, 75.88 mmol) and triethyl- amine (20.14 mL, 14.62 g, 144.50 mmol) were added consecutively. The reaction mixture was stirred for 18 hours at room temperature, poured into water (1.5 L) and stirred for an ad¬ ditional 1 hour. The crude product was filtered off and dissolved in dichloromethane (500 mL). The organic phase was washed with water (2 x 250 mL) and dried with magnesium sul¬ fate (1 h). Filtration followed by concentration yielded a solid which was triturated in heptanes (200 mL). Filtration furnished 3-[2-(triphenylmethyl)-2H-tetra2ol-5-yl]-9H-carbazole (31.5 g) which was used without further purification. 1H-NMR (CDCI3): <?8.87 (1H, d), 8.28 (1H, bs), 8.22 (1H, dd), 8.13 (1H, d), 7.49 (1H, d), 7.47- 7.19 (18H, m); HPLC-MS (Method C): m/z: 243 (triphenylmethyl); Rt = 5.72 min. 3-[2-(Triphenylmethyl)-2H-tetrazol-5-y|]-9H-carbazole (200 mg. 0.42 mmol) was dissolved in methyl sulfoxide (1.5 mL). Sodium hydride (34 mg, 60 %, 0.85 mmol) was added, and the resulting suspension was stirred for 30 min at room temperature. 3-Chlorαbenzyl chloride (85 μL, 108 mg, 0.67 mmol) was added, and the stirring was continued at 400C for 18 hours, The reaction mixture was cooled to ambient temperature and poured into 0.1 N hydrochloric acid (aq.) (15 mL). The precipitated solid was filtered off and washed with water (3 x 10 rnL) to furnish ^(S-chlorobenzyO-S-P^triphenylmethylJ^W-tetrazol-δ-yll-SH-carbazole, which was dissolved in a mixture of tetrahydrof uran and 6 N hydrochloric acid (aq.) (9:1 ) (10 mL) and stirred at room temperature for 18 hours. The reaction mixture was poured into water (100 mL). The solid was filtered off and rinsed with water (3 x 10 mL) and dichloromethane (3 x 10 mL) to yield the title compound (127 mg). No further purification was necessary. 1H-NMR (DMSO-CZ6): 58.89 (1H, d), 8.29 (1H, d), 8.12 (1H, dd), 7.90 (1H, d). 7.72 (1H, d), 7.53 (1 H, t), 7.36-7.27 (4H, m), 7.08 (1 H1 bt), 5.78 (2H, S); HPLC-MS (Method B): m/z: 360 (M+1); Rt= 5.07 min.
The compounds in the following examples were prepared in a similar fashion. Optionally, the compounds can be further purified by recrystallization from e.g. aqueous sodium hydroxide (1 N) or by chromatography.
Example 721 (General Procedure (J)). 9-(4-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000295_0001
HPLC-MS (Method C): m/z: 360 (M+1); Rt = 4.31 min.
Example 722 (General Procedure (J)). 9-(4«Methylbenzyl)-3-(2H-tetrazol-5-yl)-9tf-carbazole
Figure imgf000296_0001
HPLC-MS (Method C): m/z: 340 (M+1); Rt = 4.26 min.
Example 723 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-(4-trifluoromethyfbeπzyl)-9W-carbazote
Figure imgf000296_0002
HPLC-MS (Method C): m/z: 394 (M+1); Rt = 4.40 min.
Example 724 (General Procedure (J)). 9-(4-BenzyIoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000296_0003
HPLC-MS (Method C): m/z: 432 (M+1); Rt= 4.70 min.
Example 725 (General Procedure (J)). 9-(3-Methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000296_0004
HPLC-MS (Method C): m/z: 340 (M+1); Rt = 4.25 min. Example 726 (General Procedure (J)). 9-Benzyl-3-(2H-tetrazol-5-yl)-9tf-carbazole
Figure imgf000297_0001
1H-NMR (DMSO-dβ): J 8.91 (1H, dd). 8.30 (1H. d), 8.13 (1H, dd). 7.90 (1H, d), 7.73 (1H1 d), 7.53 (1 H, t).7.36-7.20 (6H. m), 5.77 (2H1 s).
Example 727 (General Procedure (J)). 9-(4-Phenylbenzy()-3-(2H-tetrazo!-5-yl)-9/-f-carbazole
Figure imgf000297_0002
1H-NMR (DMSO-Cf6): 58.94 (1H, s), 8.33 (1H, d), 8.17 (1H, dd), 7.95 (1H, d). 7.77 (1H, d), 7.61-7.27 (11H. m), 5.82 (2H, s).
Example 728 (General Procedure (J)). 9-{3-Methoxybenzyl)-3«(2H-tetrazαl-5-yl>9H-carbazole
Figure imgf000297_0003
HPLC-MS (Method C): m/z: 356 (M+1); Rt = 3.99 min.
Example 729 (General Procedure (J)). 9-(Naphtha!en-2-ylmethy!)-3-(2H-tetrazol-5-yl)-9H-carbazole
HPLC-MS (Method C): m/z: 376 (M+1); Rt = 4.48 min.
Example 730 (General Procedure (J)). 9-(3-Bromobenzyl)-3-(2W-tetrazol-5-yl)-9H-carbazole
HPLC-MS (Method C): m/z: 404 (M+1); Rt = 4.33 min.
Example 731 (General Procedure (J)). 9-(Biphenyl-2-ylmethyl)-3-(2H-tetrazol-5-yl)-9W-carbazole
Figure imgf000298_0003
HPLC-MS (Method C): m/z: 402 (M+1 ); Rt = 4.80 min.
Example 732 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-[4-( 1 ,2,3-trιiadiazol-4-yl)benzyl]-9H-carbazo!e
Figure imgf000298_0004
Example 733 (General Procedure (J)). 9-(2'-Cyanobiphenyl-4-ylmethyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000299_0001
1H-NMR <DMSO-d8): <Ϊ8.91 (1H. d). 8.31 (1H, d), 8.13 (1H. dd), 7.95 (1H, d), 7.92 (1H, d), 7.78 (1H, d), 7.75 (1H, dt), 7.60-7.47 (5H, m), 7.38-7.28 (3H, m), 5.86 (2H1 s); HPLC-MS (Method C): m/z: 427 (M+ 1); Rt = 4.38 min.
Example 734 (General Procedure (J)). 9-(4-JodobenzyJ)-3-(2W-tetrazol-5-yl)-9rtl-carbazole
Figure imgf000299_0002
HPLC-MS (Method C): m/z: 452 (M+ 1); Rt = 4.37 min.
Example 736 (General Procedure (J)). 9-(3,5-Bis(trifluoromethyl)benzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000299_0003
HPLC-MS (Method C): m/z: 462 (M+1); Rt = 4.70 min.
Example 736 (General Procedure (J)). 9-(4-Bromobeπzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000300_0001
1H-NMR (DMSO-dβ): S8.89 (W, d), 8.29 <1H, d), 8.11 (1H, dd), 7.88 <1H, d), 7.70 (1H. d). 7.52 (1H, t). 7.49 (2H, d), 7.31 (1H, t), 7.14 (2H, d), 5.74 (2H. s); HPLC-MS (Method C): m/z: 404 (M+ 1); Rt = 4.40 min.
Example 737 (General Procedure (J)). 9-(Anthracen-9-yImethyl)-3-(2W-tetrazol-5-yl)-9H-carba2ole
Figure imgf000300_0002
HPLC-MS (Method C): m/z: 426 (M+1); Rt = 4.78 min.
Example 738 (General Procedure (J)). 9-(4-Carboxybenzyl)-3-(2W-tetrazol-5-y^9W-carbazo!e
Figure imgf000300_0003
3.6 fold excess sodium hydride was used.
1H-NMR (DMSO-dβ): 512.89 (1H1 bs), 8.89 (1H, d), 8.30 (1H, d), 8.10 (1H, dd), 7.87 (1H, d), 7.86 (2H1 d), 7.68 (1H, d), 7.51 (1H, t). 7.32 (1H. t), 7.27 (2H1 d), 5.84 (2H1 s); HPLC-MS (Method C): m/z: 370 (M+1); Rt = 3.37 min.
Alternative mode of preparation of 9-(4-Carboxybenzyl)-3-(2H-tetrazol-5-yl)-9H- carbazole: Carbazole (52.26 g, 0.30 mol) was dissolved in dichloromethane (3 L) and silicagel (60 mesh, 600 g) was added to the mixture and the mixture was cooled to 10 0C. A mixture of N-bromosuccinimide (NBS, 55 g, 0.30 mol) in dichloromethane (400 mL) was added at 10 0C. After addition, the mixture was allowed to reach room temperature. After standing for 42 hours, the mixture was filtered, and the solid was washed with dichloromethane (4 x 200 mL), the combined filtrates were washed with water (300 mL) and dried over Na2SO4 . Evapora¬ tion in vacuo to dryness afforded 77 g of crude product. Recrystallizatϊon from 2-propanol (800 mL) afforded 71 % 3-bromocarbazole.
To a stirred solution of 3-bromocarbazole (63 g, 0.256 mol) in W-methylpyrroIidone
(900 mL) was added cuprous cyanide (CuCN1 25.22 g, 0.28 mol) and the mixture was heated to 1900C. After 9 hours of heating, the mixture was cooled to room temperature. The mixture was concentrated by bulb-to-bulb distillation (1000C1 0.1 mm Hg). The residue was treated with NH4OH (25%, 300 mL) and subsequently extracted with ethyl acetate (10%) in toluene. The organic layer was dried over Na2SO4 and concentrated by bulb-to-bulb distillation (100 0C, 0.1 mm Hg) to give 34 g (70%) of 3-cyanocarbazole.
Sodium hydride 55-60% in mineral oil (3.7 g. 0.093 mol) was added in portions to a stirred, cooled (5 0C) mixture of 3-cyanocarbazole (17.5 g, 0.091 mol) in W1W- dimethylformamide (200 mL). After 0.5 hours, a solution of methyl 4-bromomethyfbenzoate (22.9 g, 100 mmol) in W,W-dimethylformamidβ (80 mL) was added dropwise to the cooled mixture. The mixture was subsequently slowly warmed to room temperature and stirred overnight. The mixture was poured into ice water and extracted with dichloromethane (2 x 200 mL), the organic layer was washed several times with water, dried over Na2SO4 and concentrated in vacuo. A mixture of ethyl acetate and heptane (1/1 , 50 mL) was added to the concentrate and the solid was product filtered off. Yield 24 g (78%) of 4-(3-cyanocarba2θl-9~ ylmethyl)benzoic acid methyl ester.
Sodium azide (7.8 g, 0.12 mol) and ammonium chloride (6.42 g , 0.12 mol) were added to a stirred mixture of 4-{3-cyanocarbazol-9-ylmethyl)benzoic acid methyl ester (24.8 g, 0.073 mol) in WtW-dimethylformamide (130 mL) and the mixture was heated to 1100C. Af¬ ter 48 hours, the mixture was cooled to room temperature and poured into water (500 mL) and cooled to 5 0C. Hydrochloric acid (10 N) was then added to pH a 2. After stirring for 1 hour at 50C the precipitate was filtered off and washed with water. The solid obtained was air dried. Yield 27.9 g of 4-(3-(1H-tetrazo[-5-yl)carbazol-9-ylmethyl]benzoic acid methyl ester. 31,1 g of 4-[3-(iH-tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid methyl ester was added to a solution of sodium hydroxide (8.76 g, 0.219 mol) in water (150 ml_) and the mixture was heated to 800C, after 0.5 h activated carbon (0.5 g) was added and the mixture was filtered through celite. The filtrate was treated with hydrochloric acid (10 N) to pH * 1 and the formed precipitate was filtered off and air dried. This procedure was repeated as the first treatment did not give complete hydrolysis of the ester. Finally the product was dissolved in 2-propanol, the filtered the mother liquor was concentrated to approximately 100 mL and the product was isolated by filtration to afford 19 g of the title compound. After evaporation of the mother liq¬ uor to dryness and re-treatment with 2-propanol further 8 g of product was isolated resulting in a yield of 90 %.
Example 739 (General Procedure (J)). 9-(2-Chlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000302_0001
HPLC-MS (Method B): m/z: 360 (M+1); Rt = 5.30 min.
Example 740 (General Procedure (J)). 9-(4-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000302_0002
1H-NMR (DMSO-de): £8.88 (1HP d). 8.28 (1H. d). 8.10 (1H, dd), 7.89 (1H. d), 7.72 (1H, d), 7.52 (1H, t), 7.31 (1H, t), 7.31-7.08 (4H, m). 5.74 (2H, s); HPLC-MS (Method C): m/z: 344 (M+1); Rt = 4.10 min.
Example 741 (General Procedure (J)). 9-(3-Fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000303_0001
1H-NMR (DMSOd6): S8.89 (1H, d), 8.29 (1H1 d), 8.12 (1H, dd), 7.90 (1H, d), 7.72 <1H,.d), 7.53 (1H, t), 7.37-7.27 (2H, m), 7.12-7.02 (2H. m), 6.97 (1 H1 d), 5.78 (2H, s); HPLC-MS (Method C): m/z: 344 (M+ 1); Rt = 4.10 min.
Example 742 (General Procedure (J)). 9-(2-lodobenzyl)-3-(2H-tetrazol-5-yl)'9H-carbazole
Figure imgf000303_0002
HPLC-MS (Method C): m/z: 452 (M+1); Rt = 4.58 mirt.
Example 743 (General Procedure (J)). 9-(3~Carbαxybenzyl)-3-(2W-tetrazo[-5-yl)-9H-carbazole
Figure imgf000303_0003
3.6 fold excess sodium hydride was used.
1H-NMR (DMSO-d6y. δ 12.97 (1H1 bs), 8.90 (1H1 bε), 8.30 (1H, d), 8.12 (1H, bd), 7.89 (1H, d), 7.82 (1H, m), 7.77 (1H, bs), 7.71 (1H, d), 7.53 (1H, t), 7.46-7.41 (2H, m), 7.32 (1H, t), 5.84 (2H, s); HPLC-MS (Method C): m/z: 370 (M+1); Rt = 3.35 min.
Example 744 (General Procedure (J)).
9-[4-(2-Propyl)benzyl]-3-(2H-tetrazol-5-yl)-9tf-carbazole
Figure imgf000304_0001
1H-NMR (DMSO-c/β): S8.B7 (1H, d), 8.27 (1H, d), 8.10 (1H, dd), 7.87 (1H, d). 7.71 (1H, d), 7.51 (1H, t), 7.31 (1H. t), 7.15 (2H, d), 7.12 (2H, d). 5.69 (2H, s), 2.80 (1H, sept), 1.12 (6H, d); HPLC-MS (Method C): m/z: 368 (M+1); Rt = 4.73 min.
Example 745 (General Procedure (J)). 9-(3,5-Dimethoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazote
Figure imgf000304_0002
HPLC-MS (Method C): m/z: 386 (M+1); Rt = 4.03 min.
Example 746 (General Procedure (J)). 3-(2H-Tetrazo!-5-yl)-9-(2,4,5--tπfluorober)zy|)-9H-carbazo!e
Figure imgf000304_0003
HPLC-MS (Method B): m/z: 380 (M+1); Rt = 5.00 min.
Example 747 (General Procedure (J)). N-Methyl-W-phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000305_0001
HPLC-MS (Method B): m/z: 383 (M+1); Rt = 4.30 min.
Example 748 (General Procedure (J)). 9-(4-Methoxybenzyl)-3~(2tf-tetrazol-5-yl)-9H-carbazote
Figure imgf000305_0002
1H-NMR (DMSO-tfβ): £8.86 (1HV d), 8.26 (1H, d), 8.10 (1H, dd), 7,90 (1H, d}, 7.73 (1H, d), 7.51 (1H, t), 7,30 (1H, t), 7.18 (2H, d).6.84 (2H, d), 5.66 (2H1 s), 3.67 (3H, s); HPLC-MS (Method B): m/z: 356 (M+1); Rt = 4.73 min.
Example 749 (General Procedure (J)). 9-(2-Methoxybenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000305_0003
1H-NMR (DMSO-rffi): £8.87 (1H, d). 8.27 (1H. d), 8.09 (1H, dd), 7.77 (1H, d), 7.60 (1H, d). 7.49 (1H, t), 7.29 (1H1 1), 7.23 (1H, bt), 7.07 (1H1 bd), 6.74 (1H, bt), 6.61 (1H, bd), 5.65 (2H, s), 3.88 (3H. s); HPLC-MS (Method B): m/z: 356 (M+1 ); Rt = 4.97 min.
Example 750 (General Procedure (J)). 9-(4-Cyanobenzyl)-3-(2/-Metrazol-5-yl)-9H-carbazole
Figure imgf000306_0001
HPLC-MS (Method C): m/z: 351 (M+1); Rt = 3.74 min.
Example 751 (General Procedure (J)). 9-(3-Cyanobenzyl)-3-(2W-tetrazol-δ-yl)-9W-carbazofe
Figure imgf000306_0002
HPLC-MS {Method C): m/z: 351 (M+1); Rt = 3.73 min.
Example 752 (General Procedure (J)). 9-(5-Chloro-2-methoxyben2yl)-3-(2W-tetrazo!-5-yl)-9H-carbazole
Figure imgf000306_0003
'H-NMR (DMSOd6): «58.87 (1H, d), 8.35 (1H, d), 8.10 (1 H, dd), 7.73 (1H1 d), 7.59 (1H1 d), 7.49 (1H, t), 7.29 (1H, t), 7.27 (1H1 dd), 7.11 (1H1 d).6.51 (1H, d). 5.63 (2H, s), 3.88 (3H, s); HPLOMS (Method C): m/z: 390 (M+1); Rt = 4.37 min.
Example 753 (Genera) Procedure (J)). Λ/-Phenyl-2-[3-(2W-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000307_0001
1H-NMR (DMSO-Cf6): £10.54 (1H, s), 8.87 (1H1 bs), 8.27 (1H, d), 8.12 (1HT bd), 7.83 (1H. d), 7.66 (1H, d), 7.61 (2H, d), 7.53 (1H,t), 7.32 (1H, t), 7.32 (2H, t), 7.07 (1H, t), 5.36 (2H, s); HPLC-MS (Method C): m/z: 369 (M+1); Rt = 3.44 min.
Example 754 (General Procedure (J)). ΛA-Butyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetarπide
Figure imgf000307_0002
1H-NMR (DMSOd6): 58.85 (1H, d). 8.31 (1H, t), 8.25 (1H, d), 8.10 (1H, dd), 7.75 (1H, d), 7.58 (1H, d). 7.52 (1H, t), 7.30 (1H, t). 5.09 (2H, S)1 3.11 (2H1 q), 1.42 (2H1 quint). 1.30 (2H, sext), 0.87 (3H, t); HPLC-MS (Method C): m/z: 349 (M+1); Rt = 3.20 min.
Example 755 (General Procedure (J)), 9-(2,4-Dichlorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000307_0003
1H-NMR (DMSO-Cf6): <J8.92 (1H1 d), 8.32 (1H, d), 8.09 (1H. dd), 7.76 (1H, d), 7.74 (1H1 d), 7.58 (1Ht d). 7.51 (1H1 1), 7.33 (1H, t)r 7.23 (1H. dd), 6.42 (1H, d), 5.80 <2H, s); HPLC-MS (Method B): mfe: 394 (M+1); Rt = 5.87 min.
Example 756 (General Procedure (J)). 9~(2-Methylbenzyl)-3-(2W-tetrazol-5-yl)-9H-carbazole
Figure imgf000308_0001
1H-NMR (DMSO-tfβ): J8.92 (1H, d), 8.32 <1H, d), 8.08 (1H, dd), 7.72 (1H, d), 7.55 (1H, d), 7.48 (1H, t). 7.32 <1H, t), 7.26 (1H, d), 7.12 (1H, t), 6.92 (1H, t), 6.17 (1H, d), 5.73 (2H, S), 2.46 (3H, s); HPLC-MS (Method B): m/z: 340 (M+1); Rt = 5.30 min.
Example 757 (Genera! Procedure (J)). 9-(3-Nitrobenzyl)-3-(2H-tetrazQl-5-yl)-9H-carbazole
Figure imgf000308_0002
HPLC-MS (Method C): m/z: 371 (M+1); Rt = 3.78 min.
Example 758 (General Procedure (J)). 9-(3,4-Dichlorobenzyl)-3-(2H-tetrazo!-5-yl)-9H-carba2oIe
Figure imgf000308_0003
HPLC-MS (Method B): m/z; 394 (M+1); Rt ~ 5.62 min.
Example 759 (General Procedure (J)). 9-(2,4-Difluorobenzyl)-3-(2H-tetrazol-5-y1)-9H-carbazole
Figure imgf000309_0001
1H-NMR (DMSO-4,): £8.89 (1H, d), 8.29 <1H, d), 8.11 (1H, dd), 7,88 <1H, d), 7.69 (1H, d), 7.52 (1H, t), 7.36-7.24 (2H, m). 7.06-6.91 (2H1 m), 5.78 (2H1 s); HPLC-MS (Method B): m/z: 362 (M+1); Rt = 5.17 miα
Example 760 (General Procedure (J)). 9-(3.5-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9W-carbazole
Figure imgf000309_0002
1H-NMR (DMSO-CZ6): S8.90 (1H, bs), 8.31 (1H, d), 8.13 (1H, bd). 7.90 (1H1 d), 7.73 (1H, d). 7.54 (1H, t), 7.34 (1H, t), 7.14 (1H, t), 6.87 (2H, bd). 5.80 (2H. s); HPLC-MS (Method B): m/z: 362 (M+1), Rt = 5.17 min.
Example 761 (General Procedure (J)). 9-(3,4-Difluorobenzyl)-3-(2W-tetrazol-5-yl)-9W-carbazole
Figure imgf000309_0003
1H-NMR (DMSO-Cf6): 58.89 (1 H, bs), 8,29 (1H, d), 8.12 (1H1 bd), 7.92 (1H, d), 7.74 (1H, d), 7.54 (1H. t), 7.42-7.25 (3H, m), 6.97 (1H, bm), 5.75 (2H, s); HPLC-MS (Method B): m/z: 362 (M+1); Rt = 5.17 min.
Example 762 (General Procedure (J)).
9-(3-lodobenzyl)-3-(2H-tetrazoi-5-yl)-9H-carbazole
Figure imgf000310_0001
HPLC-MS (Method B): m/z: 452 (M+1); Rt = 5.50 min.
Example 763 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-[3-(trifluoromethyl)beπzyl]-9H-carbazo!e
Figure imgf000310_0002
1H-NMR (DMSO-Of6): £8.89 (1H, d), 8.30 (1H, d), 8.11 (1H, dd), 7.90 (1H1 d), 7.72 (1H, d), 7.67 (1H, bs), 7.62 (1H, bd), 7.53 (1H, t), 7.50 (1H, bt), 7.33 (1H, bd), 7.32 (1H, t). 5.87 (2H. s); HPLC-MS (Method B): m/z: 394 (M+1); Rt= 5.40 min.
Example 764 (General Procedure (J)). Λ/-(4-Carboxyphenyl)-2-{3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000310_0003
3.6 fold excess sodium hydride was used.
HPLC-MS (Method B): m/z: 413 (M+1); Rt = 3.92 min.
Example 765 (General Procedure (J)). Λ^(2-Propyl)-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetamide
Figure imgf000311_0001
HPLC-MS (Method B): m/z: 335 <M+1); Rt = 3.70 min.
Example 766 (General Procedure (J)). /V-Benzyl-W-phenyl-2-[3-(2H-tetrazol-5-yl)carbazol-9-yl]acetannide
Figure imgf000311_0002
HPLC-MS (Method B): m/z: 459 (M+1); Rt = 5.37 min.
Example 767 (General Procedure (J)). A/-[4-(2-Methyl-2-propyl)phenyl]-2-[3-(2H-tetrazol-5-yl)carbazol-9-ylJacetamide
Figure imgf000311_0003
HPLC-MS (Method B): m/z: 425 (M+1 ); Rt = 5.35 min.
Example 768 (General Procedure (J)). Λ/-Phenethyl-2-[3-(2«-tetrazol-5-yl)carba20l-9-yf]acetamtde
Figure imgf000312_0001
HPLC-MS (Method C): m/z: 397 (M+1); Rt = 3.43 min.
Example 769 (General Procedure (J)). 3-(2H-Tetrazol-5-yl)-9-[2-(trifluoromethyl)benzyl]-9H-carbazole
Figure imgf000312_0002
HPLC-MS (Method C): m/z: 394 (M+1); Rt = 4.44 min.
Example 770 (General Procedure (J)). 9-[2-Fluoro-6-(trifluoromethyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000312_0003
HPLC-MS (Method C): m/z: 412 (M+1); Rt = 4.21 min.
Example 771 (General Procedure (J)). 9-[2,4-Bis(trifluoromethyl)benzyl)]-3-(2W-tetrazol-5-yl)-9H-carbazole
Figure imgf000312_0004
HPLC-MS (Method C): m/z: 462 (M+1); Rt = 4.82 min.
Example 772 (General Procedure (J)). 3-(2H-Tetrazol-5-yO~9-(2A,6-trimethylbenzy1)-9H-carbazole
Figure imgf000313_0001
HPLC-MS (Method C): m/z: 368 (M+1); Rt = 4.59 min.
Example 773 (General Procedure (J)). 9-(2,3t5,6-Tetramethylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000313_0002
HPLC-MS (Method C): m/z: 382 (M+1); Rt = 4.47 min.
Example 774 (General Procedure (J)). 9-l(Naphtrialen-1-y])methyη-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000313_0003
HPLC-MS (Method C): m/z: 376 (M+1); Rt = 4.43 min.
Example 775 (General Procedure (J)). 9-[Bis(4-fluorophenyl)methyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000314_0001
HPLC-MS (Method C): m/z: 438 (M+1); Rt - 4.60 min.
Example 776 (General Procedure (J)). 9-(2-Bromobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000314_0002
HPLC-MS (Method C): m/z: 404 (M+1); Rt = 4.50 min.
Example 777 (General Procedure (J)). 9-(2-Fluorobenzyl)-3-(2W-tetrazol-5-yl)-9W-carbazole
Figure imgf000314_0003
HPLC-MS (Method C): m/z: 344 (M+1); Rt = 4.09 min.
Example 778 (General Procedure (J)). 9-(4-Carboxy-2-methylbenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000314_0004
In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): m/z: 384 (M+1); Rt = 3.56 min.
Example 779 (General Procedure (J)). 9-(2-Phenylethyl)-3'(2W-tetrazol-5-yl)-9H-carbazole
Figure imgf000315_0001
HPLC-MS (Method C): rn/z: 340 (M+1); Rt = 4.08 min.
Example 780 (General Procedure (J)). 9-I2-Fluoro-5-(trifluoromethyl)benzyl]-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000315_0002
HPLC-MS (Method C): m/z: 412 (M+1); Rt = 4.34 min.
Example 781 (General Procedure (J)). 9-(4-Carboxy-2-fluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000315_0003
3-Fluoro-4-methylbenzoic acid (3.Og, 19.5 mmol) and benzoyl peroxide (0.18 g, 0.74 mmol) were suspended in benzene. The mixture was purged with N2 and heated to reflux. Λf- Bromosuccinimide (3.47 g, 19.5 mmol) was added portionwise, and reflux was maintained for 18 hours. The reaction mixture was concentrated, and the residue was washed with water (20 mL) at 700C for 1 hour. The crude product was isolated by filtration and washed with ad¬ ditional water (2 x 10 mL). The dry product was recrystallized from heptanes. Filtration fur- nished 4-bromomethyl-3-fluorobenzoic acid (1.92 g) which was used in the following step ac¬ cording to General Procedure (J).
In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): m/z: 388 (M+1); Rt = 3.49 min.
Example 782 (General Procedure (J)). 5-{4-[[(3'(2H-Tetrazol-5-yl)carbazol-9-yl)methyl]naphthalen-1-yl]oxy}pentanoic Acid
Figure imgf000316_0001
5-[(4-Formylnaphthalen-1-yl)oxy]pentanoic acid intermediate obtained in example 470(3.0 g, 11.0 mmol) was dissolved in a mixture of methanol and tetrahydrofuran (9:1) (100 ml_), and sodium borohydride (1.67 g, 44.1 mmol) was added portionwise at ambient temperature. Af¬ ter 30 minutes, the reaction mixture was concentrated to 50 ml. and added to hydrochloric acid (0.1 N, 500 mL). Additional hydrochloric acid (1 N, 40 mL) was added, and 5-[(4- hydroxymethyl-naphthalen-1-yl)oxy]pentanoic acid (2.90 g) was collected by filtration. To the crude product was added concentrated hydrochloric acid (100 mL), and the suspension was stirred vigorously for 48 hours at room temperature. The crude product was filtered off and washed with water, until the pH was essentially neutral. The material was washed with-hep- tanes to furnish 5-[(4-chloromethylnaphthalen-1-yl)oxy]peπtanoic acid (3.0 g) which was used in the following step according to General Procedure (J).
In this preparation, a 3.6-fold excess of sodium hydride was used. HPLC-MS (Method C): m/z: 492 (M+1 ); Rt = 4.27 min.
Example 783 (General procedure (J)) 9-(2,3-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carbazo!e
Figure imgf000317_0001
HPLC-MS (Method C): m/z= 362 (M+1); Rt = 4.13 miα
Example 784 (General procedure (J)) 9-(2,5-Difluorobenzyl)-3-(2H-tetrazol-5-yl)-9H-carba2ole
Figure imgf000317_0002
HPLC-MS (Method C): m/z = 362 (M+1); Rt = 4.08 min.
Example 785 (General procedure (J)) 9-Pentafluorophenylmethyl-3-(2H-tetrazol-5-yl)-9H-carbazole
Figure imgf000317_0003
HPLC-MS (Method C): m/z = 416 (M+1); Rt = 4.32 min.
Example 786 (General procedure (J)) 9-(2,6-Difluorobenzyl)-3-(2H-tetrazol-5-y^9H-carbazole
Figure imgf000318_0001
HPLC-MS (Method C): m/z = 362 (M+ 1); Rt = 3.77 min.
Further compounds of the invention that may be prepared according to general procedure (J), and includes:
Example 787 Example 788 Example 789
Figure imgf000319_0001
Example 790 Example 791 Example 792
Figure imgf000319_0002
Example 793 Example 794 Example 795
Figure imgf000319_0003
Example 796 Example 797 Example 798
Figure imgf000319_0004
Example 799
Figure imgf000319_0005
The following compounds of the invention may be prepared eg. from 9-(4-bromobenzyl)-3- (2H-tetrazol-5-yl)-9H-carbazole (example 736) or from 9-(3-brornobenzyl)-3-(2/-/-tetrazol-5- yl)-9H-carbazole (example 730) and aryl boronic acids via the Suzuki coupling reaction eg as described in Littke, Dai & Fu J. Am. Chem. Soc, 2000, 122, 4020-8 (or references cited therein), or using the methodology described in general procedure (E), optionally changing the palladium catalyst to bis(tri-terf-butylphosphine)palladium (0).
Figure imgf000320_0001
General procedure (K) for preparation of compounds of general formula li0:
Figure imgf000320_0002
wherein T is as defined above.
The general procedure (K) is further illustrated by the following example: Example 806 (General procedure (K)). 1 -Benzyl-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000320_0003
5-Cyanoindole (1.0 g, 7.0 mmol) was dissolved in Λ/,Λ/-dimethylformamide (14 mL) and cooled in an ice-water bath. Sodium hydride (0.31 g, 60 %, 7.8 mmol) was added, and the resulting suspension was stirred for 30 min. Benzyl chloride (0.85 mL, 0.94 g, 7.4 mmol) was added, and the cooling was discontinued. The stirring was continued for 65 hours at room temperature. Water (150 ml.) was added, and the mixture was extracted with ethyl acetate (3 x 25 ml_). The combined organic phases were washed with brine (30 mL) and dried with so¬ dium sulfate (1 hour). Filtration and concentration yielded the crude material. Purification by flash chromatography on silica gel eluting with ethyl acetate/heptanes = 1 :3 afforded 1.60 g i-benzyl-IH-indole-5-carbonitrile.
HPLC-MS (Method C): m/z: 233 (M+1); Rt = 4.17 min.
i'Benzyl-IH-indole-5-carbαnitriIe was transformed into 1-benzyt-5-(2H-tetrazol-5-yl)-1H- indole by the method described in general procedure (J) and in example 594. Purification was done by flash chromatography on silica gel eluting with dichlorαmethane/methanol = 9:1.
HPLC-MS (Method C): m/z: 276 (M+1); Rt - 3.35 min.
The compounds in the following examples were prepared by the same procedure.
Example 807 (General procedure (K)).
1 -(4-BTOmobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000321_0001
HPLC-MS (Method C): m/z: 354 (M+1); Rt = 3.80 min.
Example 808 (General procedure (K)).
1 -(4-Phenylbenzyl)-5-(2H-tetrazol-5-y!)-1 H-indole
Figure imgf000321_0002
1H-NMR (200 MHz, DMSO-Qf6): δ = 5.52 (2H, s), 6.70 (1H, d), 7.3-7.45 (6H, m), 7.6 (4H, m), 7.7-7.8 (2H1 m), 7.85(1H1 dd), 8.35 (1H, d). Calculated for C22Hi7N5. H2O: 73.32% C; 5.03% H; 19.43% N. Found: 73.81% C; 4.90% H; 19.31% N.
Example 809 4"-[5-(2H-Tetrazol-5-yl)indol-1-ylmethyl]biphenyl-4-carboxylic acid
Figure imgf000322_0001
5-{2H-Tetrazol-5-yl)-1 H-indole (Syncom BV1 Groningen, NL) (1.66g, 8.9 mmol) was treated with trityl chloride (2.5 g, 8.9 mmol) and triethyl amine (2.5 ml_, 17.9 mmol) in DMF(25 ml_) by stirring at RT overnight. The resulting mixture was treated with water. The gel was isolated, dissolved in methanol, treated with activated carbon; filtered and evaporated to dryness in vacuo. This afforded 3.6 g (94%) of crude 5-(2-trityl-2H-tetrazol-5-yl)-1 H-indole.
HPLC-MS (Method C): m/z = 450 (M+23); Rt. = 5.32 min.
4-Methylphenylbenzoic acid (5 g, 23.5 mmol) was mixed with CCI4 (100 mL) and under an atmosphere of nitrogen, the slurry was added W-Bromosuccinimide (4.19 g, 23.55 mmol) and dibeπzoyl peroxide (0.228 g, 0.94 mmol). The mixture was subsequently heated to reflux for 0.5 hour. After cooling, DCM and water (each 30 mL) were added. The resulting precipitate was isolated, washed with water and a small amount of methanol. The solid was dried in vacuo to afford 5.27 g (77%) of 4'-bromomethylbiphenyl-4-carboxylic acid.
HPLC-MS (Method C): m/z = 291 (M+1); Rt. = 3.96 min.
5-(2-Trityl-2W-tetrazol-5-yl)-1 H-indole (3.6 g, 8.4 mmol) was dissolved in DMF (100 mL). Un¬ der nitrogen, NaH (60 % suspension in mineral oil, 34 mmol) was added slowly. 4'- Brornomethylbiphenyl-4-carboxylic acid (2.7 g, 9.2 mmol) was added over 5 minutes and the resulting slurry was heated at 40 0C for 16 hours. The mixture was poured into water (10OmL) and the precipitate was isolated by filtration and treated with THF/6N HCI (9/1) (70 mL) at room temperature for 16 hours. The mixture was subsequently evaporated to dryness in vacuo, the residue was treated with water and the solid was isolated by filtration and washed thoroughly 3 times with DCM. The solid was dissolved in hot THF (400 mL) treated with activated carbon and filtered. The filtrate was evaporated in vacuo to dryness. This af¬ forded 1.6 g (50%) of the title compound.
HPLC-MS (Method C): m/z - 396 (M+1); Rt. = 3.51 min.
Example 810 (General procedure (K)). 5-(2H-Tetrazol-5-yl)-1 H-indole
Figure imgf000323_0001
5-(2H-Tetrazol-5-y1)-1 H-indole was prepared from 5-cyanoindole according to the method described in example 594.
HPLC-MS (Method C): m/z: 186 (M+1); Rt = 1.68 min.
Example 811 (General procedure (K)). 1-Benzyl-4-(2H-tetrazol-5-yl)-1H-indole
Figure imgf000323_0002
1-Benzyl-1H-indole-4-carbonitrile was prepared from 4-cyanoindole according to the method described in example 806.
HPLC-MS (Method C): m/z: 233 (M+1); Rt = 4.24 min.
1-Benzyl-4-(2W-tetrazol-5-yl)-1 H-indole was prepared from 1-benzyl-1H-indole-4-carbonitrile according to the method described in example 594. HPLC-MS (Method C): m/z: 276 (M+1 ); Rt = 3-44 min. General procedure (L) for preparation of compounds of general formula I11:
, Br
Figure imgf000324_0001
wherein T is as defined above and
Pol- is a polystyrene resin loaded with a 2-chlorotrityl linker, graphically shown below:
Figure imgf000324_0002
This general procedure (L) is further illustrated by the following example: Example 812 (General procedure (L)). 5-(2H-Tetrazol-5-yl)-1 -[3-(trifluoromethyl)benzyl]-1 H-indole
Figure imgf000324_0003
2-Chlorotritylchloride resin (100 mg, 0.114 mmol active chloride) was swelled in dichloro- methane (2 mL) for 30 min. The solvent was drained, and a solution of 5-(2H-tetrazol-5-yl)- 1 H-indole (example 810) (63 mg, 0.34 mmol) in a mixture of tø.tø-dimethylformarriide, di- chloromethane and /V,W-di(2-propyl)ethylamine (DIPEA) (5:5:2) (1.1 mL) was added. The re¬ action mixture was shaken at room temperature for 20 hours. The solvent was removed by filtration, and the resin was washed consecutively with Λ/,Λ/-dimethylformamide (2 x 4 mL), dichlorornethane (6 x 4 mL) and methyl sulfoxide (2x 4 mL). Methyl sulfoxide (1 mL) was added, followed by the addition of a solution of lithium bis(trimethylsilyl)amide in tetrahydrofu- ran (1.0 M, 0.57 ml_, 0.57 mmol). The mixture was shaken for 30 min at room temperature, before 3-(trifluoromethyl)benzyl bromide (273 mg, 1.14 mmol) was added as a solution in methyl sulfoxide (0.2 ml_). The reaction mixture was shaken for 20 hours at room tempera¬ ture. The drained resin was washed consecutively with methyl sulfoxide (2 x 4 mL), di- chloromethane (2 x 4 mL), methanol (2 x4 mL), dichloromethane (2 x 4 mL) and tetrahydro- furan (4 mL). The resin was treated with a solution of hydrogen chloride in tetrahydrofuran, ethyl ether and ethanol = 8:1:1 (0.1 M, 3 mL) for 6 hours at room temperature. The resin was drained and the filtrate was concentrated in vacuo. The crude product was re-suspended in dichloromethane (1.5 mL) and concentrated three times to afford the title compound (35 mg). No further purification was necessary.
HPLC-MS (Method B): m/z: 344 (M+1); Rt = 4.35 min.
1H-NMR (DMSO-Cf6): 58.29 (1H1 s), 7.60 (1H, dd), 7.72 (2H. m), 7.64 (2H, bs), 7.56 (1H, t), 7.48 (1H1 d), 6.70 (1H, d). 5.62 (2H1 s).
The compounds in the following examples were prepared in a similar fashion. Optionatly, the compounds can be further purified by recrystallization or by chromatography.
Example 813 (General procedure (L)).
1 -(4-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000325_0001
HPLC-MS (Method B): m/z: 310 (M+1); Rt = 4.11 min.
Example 814 (General procedure (L)).
1 -(2-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000325_0002
HPLC-MS (Method B): m/z: 310 (M+1 ); Rt = 4.05 min.
Example 815 (General procedure (L)).
1 -(4-Methoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000326_0001
HPLC-MS (Method B): m/z: 306 (M+1); Rt = 3.68 min.
Example 816 (General procedure (L)).
1 -(4-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000326_0002
HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min.
Example 817 (General procedure (L)). 5-(2H-Tetrazol-5-yl)-1 -[4-(trif luoromethyl)benzyl]-1 H-indole
Figure imgf000326_0003
HPLC-MS (Method B): m/z: 344 (M+1); Rt = 4.18 min.
Example 818 (General procedure (L)).
1 -(3-Chlorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000327_0001
HPLC-MS (Method B): m/z: 310 (M+1); Rt = 4.01 min.
Example 819 (General procedure (L)). 1 -(3-Methylbenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000327_0002
HPLC-MS (Method B): m/z: 290 (M+1); Rt = 3.98 min.
Example 820 (General procedure (L)). 1 -(2,4-Dichlorobenzyl)-5-(2tf-tetrazol-5-yl)-1 H-indole
Figure imgf000327_0003
HPLC-MS (Method B): m/z: 344 (M+1); Rt = 4.41 min.
Example 821 (General procedure (L)). 1 -(3-Methoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000327_0004
HPLC-MS (Method B): m/z: 306 (M+1); Rt = 3.64 min. Example 822 (General procedure (L)). 1-(4-Fluorobenzyl)-5-(2H-tetrazol-5-yl)-1Wndole
Figure imgf000328_0001
HPLC-MS (Method B): m/z: 294 (M+1 ); Rt = 3.71 min.
Example 823 (General procedure (L)).
1 -(3-Fluorobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000328_0002
HPLC-MS (Method B): m/z: 294 (M+1 ); Rt = 3.68 min.
Example 824 (General procedure (L)).
1 -(2-lodobeπzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000328_0003
HPLC-MS (Method B): m/z: 402 (M+1); Rt = 4.11 min.
Example 825 (General procedure (L)).
1 -[(Naphthalen-2-yl)methyl]-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000328_0004
HPLC-MS (Method B): m/z: 326 (M+1); Rt = 4.18 min.
Example 826 (General procedure (L)).
1 -(3-Bromobenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000329_0001
HPLC-MS (Method B): m/z: 354 (M+1); Rt = 4.08 min.
Example 827 (General procedure (L)).
1 -(4-Carboxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000329_0002
In this preparation, a larger excess of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0
M, 1.7 mL, 1.7 mmol) was used.
HPLC-MS (Method B): m/z: 320 (M+1); Rt = 2.84 min.
Example 828 (General procedure (L)).
1 -(3-Carboxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000329_0003
In this preparation, a larger excess of lithium bis(trimethylsilyl)amide in tetrahydrofuran (1.0 M, 1.7 mL, 1.7 mmol) was used. HPLC-MS (Method B): m/z: 320 (M+1); Rt = 2.91 min. Example 829 (General procedure (L)).
1-(2,4-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1H-indole
Figure imgf000330_0001
HPLC-MS (Method B): m/z: 312 (M+1 ); Rt = 3.78 miπ.
Example 830 (General procedure (L)).
1 -(3,5-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 W-indole
Figure imgf000330_0002
HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.78 min.
Example 831 (General procedure (L)).
1 -(3,4-Difluorobenzyl)-5-(2H-tetrazol-5-yl)-1 W-indole
Figure imgf000330_0003
HPLC-MS (Method B): m/z: 312 (M+1); Rt = 3.81 min.
Example 832 (General procedure (L)).
1 -[4-(2-Propyl)benzyl]-5-(2H-tetrazol-5-yl)-1 H-iπdole
Figure imgf000331_0001
HPLC-MS (Method B): m/z: 318 (M+1); Rt = 4.61 min.
Example 833 (General procedure (L)). 1 -(3,5-Dimethoxybenzyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000331_0002
HPLC-MS (Method B): m/z: 336 (M+1 ); Rt = 3.68 min.
Example 834 (General procedure (L)). 1 -(2'-Cyanobiphenyl-4-ylmethyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000331_0003
HPLC-MS (Method B): m/z: 377 (M+1); Rt = 4.11 min.
Example 835 (General procedure (L)).
1 -(2-Methylbenzyl)-5-(2W-tetrazol-5-yl)-1 H-indole
Figure imgf000331_0004
HPLC-MS (Method B): m/z: 290 (M+ 1); Rt = 3.98 min.
Further compounds of the invention that may be prepared according to general procedure (K) and/or (L) includes:
Figure imgf000333_0001
Figure imgf000334_0001
The following compounds of the invention may be prepared eg. from 1-(4-bromobenzyl)-5- (2H-tetrazol-5-yl)-1H-indole (example 807) or from the analogue 1-(3-bromobeπzyl)-5-(2H- tetrazol-5-yl)-1 tf-indole and aryl boronic acids via the Suzuki coupling reaction eg as de¬ scribed in Littke, Dai St Fu J. Am. Chem. Soc, 2000, 122, 4020-8 (or references cited therein), or using the methodology described in general procedure (E), optionally changing the palladium catalyst to bis(tri-tert-butylphosphiπe)palladium (0).
Figure imgf000334_0002
General procedure (M) for preparation of compounds of general formula I12:
Figure imgf000334_0003
"12 wherein T is as defined above. The general procedure (M) is further illustrated by the following example: Example 865 (General procedure (M)). 1-Benzoyl-5-(2H-tetrazol-5-yl)-1H-indole
Figure imgf000335_0001
To a solution of 5-cyanoindole (1.0 g, 7.0 mmol) in dichloromethane (8 mL) was added A- (dimethylamino)pyridine (0.171 g, 1.4 mmol), triethylamine (1.96 mL, 1.42 g, 14 mmol) and benzoyl chloride (0.89 mL, 1.08 g, 7.7 mmol). The resulting mixture was stirred for 18 hours at room temperature. The mixture was diluted with dichloromethane (80 mL) and washed consecutively with a saturated solution of sodium hydrogencarbonate (40 mL) and brine (40 mL). The organic phase was dried with magnesium sulfate (1 hour). Filtration and concentra¬ tion furnished the crude material which was purified by flash chromatography on silica gel, eluting with ethyl acetate/heptanes = 2:3. i-Benzoyl-IW-indole-5-carbonitrile was obtained as a solid.
HPLC-MS (Method C): m/z: 247 (M+1 ); Rt = 4.07 min.
i-Benzoyl-IH-indole-5-carbonitrile was transformed into 1-beπzoyl-5-(2H-tetrazol-5-yl)-1H- indole by the method described in example 594.
HPLC (Method C): Rt = 1.68 min.
The compound in the following example was prepared by the same procedure.
Example 866 (General procedure (M)). 1 -BenzoyM-(2tf-tetrazol-5-yl)-1 H-indole
Figure imgf000335_0002
i-Benzoyl-IH-indoIe-4-carbonitrile was prepared from 4-cyanoindole according to the method described in example 865-
HPLC-MS (Method C): m/z: 247 (M+1); Rt = 4.24 min.
1 -Benzoyl-4-(2H~tetrazol-5-yl)-1 tf-indole was prepared from 1 -benzoyl-1 W-iπdole-4- carbonitrile according to the method described in example 594.
HPLC (Method C): Rt = 1.56 min.
Example 867 (General procedure (M))
(2-Fluoro-3-trifluoromethylphenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000336_0001
HPLC-MS (Method B): mfe = 376 {M+1 ); Rt = 4.32 min.
Example 868 (General procedure (M))
(4-Methoxyphenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-mefhanone
Figure imgf000336_0002
HPLC-MS (Method B): m/z «= 320 (M+1); Rt = 3.70 min.
Example 869 (General procedure (M))
(3-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000337_0001
HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.72 min.
Example 870 (General procedure (M)) (4-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone
Figure imgf000337_0002
HPLC-MS (Method B): m/z = 335 (M+1 ); Rt = 3.71 min.
Example 871 (General procedure (M)) NaphthaIen-2-yl-[5-(2H-tetrazol-&-yl)-iπdol-1-ylJ-methanone
Figure imgf000337_0003
HPLC-MS (Method C): m/z = 340 (M+1 ); Rt = 4.25 min.
Example 872 (General procedure (M)) (2,3-DifluorophenylH5-(2H-tetrazol-5-ylHndol-1-yl]-methanone
Figure imgf000337_0004
HPLC-MS (Method B: m/z = 326 (M+1 ); Rt = 3.85 min. The following known and commercially available compounds do atl bind to the His B10 Zn2* site of the insulin hexamer:
Example 873
1 -(4-Fluαrophenyl)-5-(2H-tetrazol-5-yl)-1 H-indole
Figure imgf000338_0001
Example 874 1-Amino-3-(2H-tetrazol-5-yl)benzene
Figure imgf000338_0002
Example 875 1-Amiπo-4-(2H-tetrazol-5-yl)benzene
Figure imgf000338_0003
A mixture of 4-aminobenzonitrile (10 g, 84.6 mmol), sodium azide (16.5 g, 254 mmol) and ammonium chloride (13.6 g, 254 mmol) in DMF was heated at 1250C for 16 hours. The cooled mixture was filtered and the filtrate was concentrated in vacuo. The residue was added water (200 mL) and diethyl ether (200 mL) which resulted in crystallisation. The mix¬ ture was filtered and the solid was dried in vacuo at 400C for 16 hours to afford 5-(4- aminophenyl)-2H-tetrazo!e.
1H NMR DMSO-dβ): δ = 5.7 (3H, bs), 6.69 (2H, d), 7.69 (2H, d). HPLC-MS (Method C): m/z: 162 (M+1); Rt = 0,55 min.
Example 8761 -Nitro-4-(2H-tetrazol-5-yl)benzene
Figure imgf000339_0001
Example 8771 -Bromo-4-(2H-tetrazol-5-yl)beπzene
Figure imgf000339_0002
General procedure (N) for solution phase preparation of amides of general formula I13:
θ ^v θ
Frag— U-OH + HN R *. p^g-U-N^R
R' R.
'» wherein Frag is any fragment carrying a carboxylic acid group, R is hydrogen, optionally sub¬ stituted aryl or C^-alkyl and R* is hydrogen or C1-4-alkyl.
Frag-CO2H may be prepared eg by general procedure (D) or by other similar procedures de¬ scribed herein, or may be commercially available.
The procedure is further illustrated in the following example 878:
Example 878 (General procedure (N))
W-(4-Ch!orobenzyl)-2-[3-(2T4-dioxothiazolidin-5-ylidenemethyl)-1H-indol-1-y|]acetamide
Figure imgf000339_0003
[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-yl]acetic acid (example 478, 90.7 mg, 0.3 mmol) was dissolved in NMP (1 mL) and added to a mixture of 1-ethyl-3-(3-dimethylamino- propyl)carbodiimide, hydrochloride (86.4 mg, 0.45 mmol) and 1-hydroxybeπzotriazol (68.8 mg, 0.45 mmol) in NMP (1 mL). The resulting mixture was shaken at RT for 2 h. 4- Chlorobenzylamine (51 mg, 0.36 mmol) and DIPEA (46.4 mg, 0.36 mmol) in NMP (1 mL) were added to the mixture and the resulting mixture shaken at RT for 2 days. Subsequently ethyl acetate (10 mL) was added and the resulting mixture washed with 2x10 mL water fol¬ lowed by saturated ammonium chloride (5 mL). The organic phase was evaporated to dry¬ ness giving 75 mg (57%) of the title compound.
HPLC-MS (Method C): m/z: 426 (M+1); Rt. = 3.79 min.
Example 879 (General procedure (N))
W-(4-Chlorobenzyl)-4-[2-chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide
Figure imgf000340_0001
HPLC-MS (Method A): m/z: 465 (M+1); Rt = 4.35 min.
Example 880 (General procedure (N)) N-(4-Chlorobenzyl)-4-j;4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyramide
Figure imgf000340_0002
HPLC-MS (Method A): m/z: 431 (M+1); Rt = 3.68 min.
Example 881 (General procedure (N)) 2-[2-Bromo-4-(2.4-dioxothiazolidin-5-ylidenemethyl)phenoxy]-W-(4-chlorobenzyl)acetamide
Figure imgf000340_0003
HPLC-MS (Method A): m/z: 483 (M+1); Rt = 4.06 min. Example 882 (General procedure (N)) Λ/-(4-ChIorobenzyl)-2-[3-(2,4-dioxothia∑olidin-5-ylidenemethyl)pheπoxy]acetamide
Figure imgf000341_0001
HPLC-MS (Method A): m/z: 403 (M+1); Rt = 4.03 min.
EΞxample 883 (General procedure (N))
W-(4-Chlorobenzyl)-3-t4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenyl]acrylamide
Figure imgf000341_0002
HPLC-MS (Method A): m/z: 399 (M+1); Rt = 3.82.
Example 884 (General procedure (N)) Λ/-(4-Chloroben2yl)-4-[3-(2.4-dioxothiazolidin-5-ylidenemetriyl)pheπoxylbutyramide
Figure imgf000341_0003
HPLC-MS (Method A): m/z: 431 (M+1); Rt = 3.84 min.
Example 885 (General procedure (N)) 4-t2-Bromo-4-(2,4-dioxothiazoIidin-5-ylidenemethyl)phenoxy]-W-(4-chlorobenzyl)butyramide
Figure imgf000341_0004
HPLC-MS (Method A): m/z: 511 (M+1); Rt = 4.05 min.
Example 886 (General procedure (N))
4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)-phenoxy]-W-(4-chlorobenzyl)- butyramide
Figure imgf000341_0005
HPLC-MS (Method A): m/z: 527 (M+ 1); Rt = 4.77 min.
Example 887 (General procedure (N)) N-(4-Chlorobenzyl)-2-[4-(2,4^ioχothiazolidin-5-ylidenernethyl)naphthalen-1-yloxy]acetamide
Figure imgf000342_0001
HPLC-MS (Method C): m/z: 431 (M+1); Rt. = 4.03 min.
Example 888 (General procedure (N)) Λ/-(4-Chlorobenzyl)-3-[3-(2,4-dioxothiazotidin-5-ylidenemethyi)-1H-indo!-1-yl]propionamide
Figure imgf000342_0002
HPLC-MS (Method C): m/z: 440 (M+1); Rt. = 3.57 min.
Example 889 (General procedure (N)) W-(4-Chlorobenzyl)-4-[4-(2,4-dioxothiazoiidin-5-ylidenemethyl)naphthalen-1-yloxy]butyramide
Figure imgf000342_0003
HPLC-MS (Method C): m/z: 481 (M+1); Rt = 4.08 min.
Example 890 (General procedure (N)) 4-[4-(2,4-Dioxothiazolidin-5-ylidenemethy!)-naphthaIen-1-yloxy]-Λ/-hexylbutyramide
Figure imgf000342_0004
HPLC-MS (Method C): m/z: 441 (M+1); Rt = 4.31 min. Example 891 (General Procedure (N))
4-({[3-(214-Dioxothia2θlidin-5-ylidenernethyl)indole-7-carbonyl]arniπo}rnethyl)ben2oic acid methyl ester
Figure imgf000343_0001
Rt= 3.55 min.
Example 892 (General procedure (N)) Λ/-(4-Chlorobenzyl)-4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzamide
Figure imgf000343_0002
HPLC-MS (Method C): m/z:493 (M+1); Rt = 4.19 min.
Example 893 (General procedure (N))
Λ/-(4-Chlorobenzyl)-3-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzamide
Figure imgf000343_0003
HPLC-MS (Method C): m/z: 493 (M+1 ); Rt = 4.20 min.
Example 894 (General Procedure (N)) Λ/-(4-Chtorobenzyl)-3-methyl-4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]benzamide
Figure imgf000344_0001
HPLC-MS (Method C): m/z: 507 (M+1); Rt = 4.37min.
Example 895 (General procedure (N)) 5-{2-[4-(2.4-Dioxothiazoltdin-5-ylidenemethyl)-naphthalen-1-yloxy]-acetylamino}-isophthalic acid dimethyl ester
Figure imgf000344_0002
HPLC-MS (Method C): m/z = 521 (M+1); Rt. = 4.57 min.
Example 896 (General procedure (N))
5-{2-[4-(2ADioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-acetylarnino}-isophthalic acid
Figure imgf000344_0003
HPLC-MS (Method C): m/z = 515 (M+23); Rt. = 3.09 min.
Example 897 (General procedure (N)) 5-(3-{2-[4-(2,4-Dioxothiaa:olidin-S-ylidenemethyl)-naphthaIen-1-yloxyJ-ethyl}-ureido)- isophthalic acid monomethyl ester
Figure imgf000345_0001
HPLC-MS (Method C): m/z = 536 (M+1); Rt = 3,58 min.
Example 898 (General Procedure (N)). 2-{4-[3-(2H-Tetrazol-5-yl)carbazot-9-ylmethyl]benzoylamino}succinic acid dimethyl ester
Figure imgf000345_0002
4-[3-(iH-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid (2.00 g, 5.41 mmol), 1- hydroxybenzotriazole (1.46 g. 10.8 mmol) and N,N-di(2-propyl)ethylamine (4.72 mL, 3.50 g,
27.1 mmol) were dissolved in dry N.N-dimethylformamide (60 mL). The mixture was cooled in an ice-water bath, and 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride
(1.45 g, 7.56 mmol) and (S)-aminosuccinic acid dimethyl ester hydrochloride (1.28 g, 6.48 mmol) were added. The cooling was discontinued, and the reaction mixture was stirred at room temperature for 18 hours before it was poured into hydrochloric acid (0.1 N, 600 mL).
The solid was collected by filtration and washed with water (2 X 25 mL) to furnish the title compound.
HPLC-MS (Method C): m/z: 513 (M+1); Rt= 3.65 min.
1H-NMR (DMSO-d6): δ 8.90 (1H1 d). 8.86 (1H, d), 8.29 (1H, d), 8.11 (1H, dd), 7.87 (1H. d), 7.75 (2H1 d), 7.69 (1H. d), 7.51 (1H, t). 7.32 (1H1 1), 7.28 (2H, d), 5.82 (2H, s), 4.79 (1H, m),
3.61 (3H, S)1 3.58 (3H, s), 2.92 (1H, dd). 2.78 (1H, dd).
Example 899 (General Procedure (N)). 2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid
Figure imgf000346_0001
2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester
(1.20 g, 2.34 mmol) was dissolved in tetrahydrofuran (30 mL). Aqueous sodium hydroxide (1
N, 14 mL) was added, and the resulting mixture was stirred at room temperature for 18 hours. The reaction mixture was poured into hydrochloric acid (0.1 N. 500 mL). The solid was collected by filtration and washed with water (2 X 25 mL) and diethyl ether (2 X 25 mL) to furnish the title compound.
HPLC-MS (Method C): m/z: 485 (M+ 1); Rt = 2.94 min.
1H-NMR (DMSO-dβ): δ 12.44 (2H, s (br)), 8.90 (1H, d). 8.68 (1H, d), 8.29 (1H, d), 8.11 (1H, dd). 7.87 (1H, d), 7.75 (2H, d), 7.68 (1H, d), 7.52 (1H1 1), 7.32 (1H, t), 7.27 (2H, d), 5.82 (2H,
S), 4.70 (1H1 m), 2.81 (1H, dd). 2.65 (1H, dd).
The compounds in the following examples were prepared in a similar fashion.
Example 900 (General procedure (N))
2-{4-[3-(2H-Tetrazol-5-yl)-carbazol-9-yfmethylJ-benzoylarnino}-succinic acid dimethyl ester
Figure imgf000346_0002
HPLC-MS (Method C): m/z = 513 (M+1); Rt = 3.65min.
Example 901 (General procedure (N))
2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethy|]benzoylamino}pentanedioic acid dimethyl ester
Figure imgf000347_0001
HPLC-MS (Method C): m/z = 527 (M+ 1); Rt = 3.57min.
Example 902 (General procedure (N)) (Methoxycarbonylmethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl}-amino)-acetic acid methyl ester
Figure imgf000347_0002
HPLC-MS (Method C): m/z = 513 (M+1); Rt = 3,55min.
Example 903 (General procedure (N))
2-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentaπedioic acid
Figure imgf000347_0003
HPLC-MS (Method C): m/z = 499 (M+1); Rt = 2.87min.
Example 904 (General procedure (N))
(Ethoxycarbonylmethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoyl}-amino)-acetic acid ethyl ester
Figure imgf000348_0001
HPLC-MS (Method C): m/z = 541 (M+1); Rt = 3.91 min.
Example 905 (General procedure (N)) 3-(3-{4-[4-(2,4-Dioxo-thiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyrylamino}- propylamino)-hexanedioic acid dimethyl ester
Figure imgf000348_0002
HPLC-MS (Method C: m/z = 585 (M+1); Rt = 2.81 min.
Example 906 (General procedure (N))
3_(3^4-[4-(2,4-Dioxo-thiazolidin-5-ylideπemethyl)-naphthalen-1-yloxy]-butyrylamino}- propylamino)-hexanediojc acid
Figure imgf000348_0003
HPLC-MS (Method C): m/z = 554 (M-3); Rt = 3,19 min.
Example 907 (General procedure (N)) (Garboxymethyl-{4-[3-(2H-tetrazol-5-yl)-carbazol-9-ylmethyη-benzoyl}-amino)-acetic acid
Figure imgf000348_0004
HPLC-MS (Method C): m/z = 485 (M+1); Rt = 3.04 min. Example 908 (General procedure (N))
4-(3-{4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)-naphthalen-1-yloxy]-butyrylamino}- propylamino)-cyc!ohexane-1 ,3-dicarboxylic acid dimethyl ester
Figure imgf000349_0001
HPLC-MS (Method C): mfz = 612 (M+1 ); Rt = 3,24 min.
Example 909 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyllbenzoylamino}pentanedioic acid dimethyl ester
Figure imgf000349_0002
HPLC-MS (Method C): m/z = 527 (M+1 ); Rt = 3.65min.
Example 910 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyIamino}pentanedioic acid dimethyl ester
Figure imgf000349_0003
HPLC-MS (Method C): m/z = 527 (M+1 ); Rt = 3.65min.
Example 911 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid dimethyl ester
Figure imgf000349_0004
HPLC-MS (Method C): m/z = 527 (M+1); Rt = 3.65min.
Example 912 (General procedure (N)) 2-{3-t3-(2H-Tetrazol-5-yl)carbazoI-9-ylmethyl]benzoylarπino}pentanedioic acid
Figure imgf000350_0001
HPLC-MS (Method C): m/z = 499 (M+1); Rt = 3.00 rran.
Example 913 (Genera! procedure (N))
(Methoxycarbonylmethyl-{3-[3-(2H-tetra2ot-5-yl)carbazol-9-ylmethyl]benzoyl}amino)acetic acid methyl ester
Figure imgf000350_0002
1H-NMR (bMSO-dβ): δ 8.88 (1H, d), 8.29 (1H, d), 8.10 (1H, dd). 7.85 (1H/d), 7.67 (1H, d), 7.52 (1H, t). 7.39 (1H. t), 7.30 (2H, m), 7.17 (2H. m). 5.79 (2H1 s). 4.17 (2H, s), 4.02 (2H, s), 3.62 (3H, s). 3.49 (3H, s).
Example 914 {General procedure (N)) 2-{3-[3-(2H-Tetra2ol-5-yl)carbazol-9-ylmethyl]benzoylamino}succinic acid dimethyl ester
Figure imgf000350_0003
HPLC-MS (Method C): m/z = 513 (M+1 ); Rt = 3.70 min.
Example 915 (General procedure (N)) 2-{3-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamiπo}-succinic acid
Figure imgf000351_0001
HPLC-MS (Method C): m/z = 485 (M+1); Rt = 2.96 min.
Example 916 (General procedure (N)) (Carboxymethyl-{3-[3-(2H-tetrazoU5-yl)carbazol-9-y1methyqbeπzoyl}amino)acetic add
Figure imgf000351_0002
HPLC-MS (Method C): m/z = 485 (M+1); Rt = 2.87 min.
Example 917 (General procedure (N)) 4-(4-(3-Carboxy-propylcarbamoyl)4-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl3- benzoylamino}-butyryla mino)-butyric acid
Figure imgf000351_0003
The title compound was prepared by coupling of (S)-2-{4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethyl]beπzoylamiπo}pentanedioic acid bis-(2,5"dioxopyrrolidin-1-yl) ester (prepared from (S)-2-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}pentanedioic acid by essen- tially the same procedure as described for the synthesis of 4-[3-(2H-tetrazol-5-yl)carbazol-9- ylmethytjbenzoic acid 2,5-dioxopyrrolidin-i-yl ester) with 4-aminobutyric acid according to the procedure described for the preparation of 4-{4-[3-(2H-tetrazol-5-yl)carbazol-9-ylmethyl]- benzoylamino}butyric acid .
HPLC-MS (Method C): m/z: 669 (M+1); Rt = 2.84 min.
Example 918 (General procedure (N)) [2-(2-{4-[3-(2H-Tetrazol-5-yl)-caιi3azol-9-ylmethyl]benzoyϊarnino}ethoxy)ethoxy]acetic acid
Figure imgf000352_0001
HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.10 min.
Example 919 (General procedure (N))
2-{4-[3-(2H-Tetrazol-5-yl)-carbazol-9-ylmethyl]-benzoylamino}-pentanedioic acid di-tert-butyl ester
Figure imgf000352_0002
HPLC-MS (Method C): m/z = 611 (M+1); Rt = 4.64 min.
Example 920 (General Procedure (N)). 4-{4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoylamino}butyricAcid
Figure imgf000352_0003
HPLC-MS (Method C): m/z: 455 (M+1); Rt = 3.13 min.
Example 921 (General Procedure (N)). [2-(2-{4-[3-(2H-Tetrazol-5-yI)carbazol-9-ylrnethyllhenzoylamino}ethoxv)ethoxylacetic aci(l
Figure imgf000353_0001
The title compound was prepared by coupling of 4-[3-(2H-tetrazol-5-yl)carbazo!-9- ylmethyljbenzoic acid 2,5-dioxopyrrolidin-1-yl ester with [2-(2-aminoethoxy)ethoxy]acetic acid (prepared from [2-[2-(Fmoo-amino)etrioxy]ethoxy]acetic acid by treatment with PS-Trisamine resin in DMF). HPLC-MS (Method C): m/z: 515 (M+1); Rt = 3.10 min.
The commercially available compounds in the following examples do all bind to the HisB10 Znz+site:
Example 922
1 -(4-Bromo-3-methylphenyI)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000353_0002
Example 923 1 -(4-lodophenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000353_0003
Example 924 1-(2,4t5-Trichlorophβnyl)-1H-tetrazole-5-thiol
Figure imgf000354_0001
Example 925
1 -(2,6-Dimethylphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000354_0002
Example 926
1 -(2,4.6-Trimethylphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000354_0003
Example 927
1 -(4-Dimethy1aminophenyl)-1 H-tetrazole-5-thiol
Figure imgf000354_0004
Example 928
1 -(3,4-Dichlorophenyl)-1 ,4-dihydro-1 W-tetrazole-5-thione
Figure imgf000354_0005
Example 929 1 -(4-Propylphenyl)-1 ,4-dihydro-1 H-tetrazole-5-thione
Figure imgf000355_0001
Example 930
1 -(3-Chloropheπyl>1 ,4-dihydro-1 H-tetrazoie-5-thione
Figure imgf000355_0002
Example 931
1 -(2-Fluorophenyl)-1 ,4-dihydro-1 H-tetrazole-5-thione
Figure imgf000355_0003
Example 932 1-(2,4-Dichlorophenyl)-1.4-dihydro-1H-tetrazole-5-thione
Figure imgf000355_0004
Example 933
^^-TrifluoromethoxyphenyD-i ^-dihydro-iH-tetrazole-S-thione
Figure imgf000355_0005
Example 934 M-[4-(5-Mercaptotetrazol-1-yl)-phenyll-acetamide
Figure imgf000356_0001
Example 935
1 -{4-Ch1orophenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000356_0002
ϊΞxample 936
1 -{4-Methoxyphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000356_0003
Example 937
1 -(3-Fluoro-4-pyrrolidin-1 -ylphenyl)-1 ,4-dihydrotetrazole-5-thione
Figure imgf000356_0004
Example 938
N-[3-(5-Merraptotetrazol-1-yl)phenyl]acetamide
Figure imgf000356_0005
Example 939 1-(4-Hydroxyphenyl)-5-mercaptotetrazole
Figure imgf000357_0001
Example 940
Figure imgf000357_0002
Preparation of 1 -aryl-1 ,4-dihydrotetrazole-5-thiones (or the tautomeric 1-aryltetrazole-5- thiols) is described in the literature (eg. by Kauer & Sheppard, J. Org. Chem., 32, 3580-92 (1967)) and is generally performed eg. by reaction of aryl-isothiocyanates with sodium azide followed by acidification
1 -Aryl-1 ,4-dihydrotetrazole-5-thiones with a carboxylic acid tethered to the aryl group may be prepared as shown in the following scheme:
Figure imgf000357_0003
Step 3
Figure imgf000357_0004
Step 1 is a phenol alkylation and is very similar to steps 1 and 2 of general procedure (D) and may also be prepared similarly as described in example 481. Step 2 is a reduction of the nitro group. SnCI2, H2 over Pd/C and many other procedures known to those skilled in the art may be utilised.
Step 3 is formation of an arylisothiocyanate from the corresponding aniline. As reagents CS2, CSCI2, or other reagents known to those skilled in the art, may be utilised.
Step 4 is a conversion to mercaptotetrazole as described above.
Compounds of the invention include:
Figure imgf000358_0001
Example 948
4-(4-Hydroxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000359_0001
Phenylsulphonyl acetonitrile (2.0 g, 11.04 mmol) was mixed with 4-hydroxybenzaldehyde (1.35 g, 11.04 mmol) in DMF (10 ml_) and toluene (20 mL). The mixture was refluxed for 3 hours and subsequently evaporated to dryness in vacuo. The residue was treated with di- ethyl ether and toluene. The solid formed was filtered to afford 2.08 g (66%) of 2- benzenesulfonyJ-3-(4-hydroxyphenyl)acrylonitrile. HPLC-MS (Method C): m/z: 286 (M+1); Rt. = 3.56 min.
A mixture of 2-benzenesulfonyl-3-(4-hydroxyphenyl)acrylonitrite (2.08 g, 7.3 mmol) and so- dium azide (0.47g,7.3 mmol) in DMF (50 mL) was heated at reflux temperature 2 hours. After cooling, the mixture was poured on ice. The mixture was evaporated in vacuo to almost dry¬ ness and toluene was added. After filtration, the organic phase was evaporated in vacuo. The residue was purified by silicagel chromatography eluting with a mixture of ethyl acetate and heptane (1 :2). This afforded 1.2 g (76%) of the title compound.
1H NMR (DMSO-cfe): 10.2 (broad.iH); 7.74 (d,2H); 6.99 (d,2H); 3.6-3.2 (broad,1H). HPLC-MS (Method C) m/z: = 187 (M+1); Rt. = 1.93 min
General procedure (O) for preparation of compounds of general formula I14:
Figure imgf000359_0002
14
wherein
AA is as defined above,
Steps 1 and 2 are described in the literature (eg Beck & Gύnther, Chem. Ber., 106, 2758-66 (1973)) Step 1 is a Knoevenagel condensation of the aldehyde AA-CHO with phenylsulfonyl- acetonitrile and step 2 is a reaction of the vinylsulfonyl compound obtained in step 1 with so¬ dium azide. This reaction is usually performed in DMF at 90 - 1100C.
This general procedure is further illustrated in the following example 949:
Example 949 (General Procedure (O)) [4-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)phenoxy]acetic acid
Figure imgf000360_0001
Phenylsulphonylacetonitrile (0.1 g, 0,55 mmol) was mixed with 4-formyiphenoxyactic acid
(0.099 g, 0.55 mmol) in DMF (3 mL) and heated to 110 0C for 3 h and subsequently cooled to RT. Sodium azide (0.036 g, 0.55 mmol) was added and the resulting mixture was heated to 1100C for 3 h and cooled to RT. The mixture was poured into water (20 mL) and centrifuged. The supernatant was discarded, ethanol (5 mL) was added and the mixture was centrifuged again. After discarding the supernatant, the residue was dried in vacuo to afford 50 mg (37%) of f4-(5-Cyano-1W-[1,2,3]triazoi-4-yl)phenoxy]acetic acid.
HPLC-MS (Method C): m/z: 245 (M+1) Rt. 2.19 min.
Example 950 (General Procedure (O))
5-(Naphthalen-1 -yl)-3W-[1 ,2,3]triazole-4-carboπitrile
Figure imgf000360_0002
HPLC-MS (Method C): m/z: 221 (M+1); Rt. 3,43 min.
Example 951 (General Procedure (O))
5-(Naphthalen-2-yl)-3H-[1f2,3]triazole-4-carbonitrile
Figure imgf000361_0001
HPLC-MS (Method C): m/z: 221 (M+1); Rt = 3.66 min.
Example 952 (Genera! procedure (O)) 4-[3-(5-Cyano-[1 ,2,3ltriazol-4-yl)-1 ^-dimethylcarbazol-θ-ylmethylj-benzoic acid
Figure imgf000361_0002
HPLC-MS (Method C): m/z = 422 (M+1); Rt= 3.85 min.
Preparation of intermediary aldehyde: 1,4 Dimethylcarbazol-3-carba(dehyde (0.68 g, 3.08 mmol) was dissolved in dry DMF (15 mL), NaH (diethyl ether washed) (0.162 g, 6.7 mol) was slowly added under nitrogen and the mix¬ ture was stirred for 1 hour at room temperature. 4-Bromomethylbenzαic acid (0.73 g, 3.4 mmol) was slowly added and the resulting slurry was heated to 40 0C for 16 hours. Water (5 mL) and hydrochloric acid (6Nt 3 mL) were added. After stirring for 20 min at room tempera- ture, the precipitate was filtered off and washed twice with acetone to afford after drying 0.38 g (34%) of 4-(3-formyl-1 ,4-dimethylcarbazol-9-ylmethyl)benzoic acid.
HPLC-MS (Method C) : m/z = 358 (M+1), RT. = 4.15 min.
Example 953 (General Procedure (O))
5-(Anthracen-9-yl)-3H-[1,2,3]triazole-4-carbonitrile
Figure imgf000362_0001
HPLC-MS (Method C): m/z: 271 (M+1); Rt = 3.87 min.
Example 954 (General Procedure (O)) 5-(4-Methoxynaphthalen-1 -yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000362_0002
HPLC-MS (Method C): m/z: 251 (M+1); Rt = 3.57 min.
Example 955 (General Procedure (O)) 5-(1 ,4-Dimethyl-9H-carbazol-3-yl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000362_0003
HPLC-MS (Method C): m/z: 288 (M+1); Rt = 3.67 min.
Example 956 (General procedure (O)) 5-(4'-Methoxybiphenyl-4-yl)-3H-[1 ,2,3]triazoIe-4-carbonitrile
Figure imgf000363_0001
HPLC-MS (Method C): m/z = 277 (M+1 ); Rt = 3.60 min.
Example 957 (General procedure (O)) 5-(4-Styrylphenyl)-3H-[1 ,2.3]triazole-4-carbonitrile
Figure imgf000363_0002
HPLC-MS (Method C): m/z = 273 (M+1); Rt » 4.12 min.
Example 958 (General procedure (O)) 5-(2,6-Dichloro-4-dibenzylaminophenyl)-3H-[1 ,2,3]triazole-4-carbonitrile
Figure imgf000363_0003
HPLC-MS (Method C): m/z = 434 (M+1 ); Rt = 4.64 min. Example 959 (General procedure (O)) 6-(1-Bromonaphthalen-2-yl)-3H-[1,2,3]triazole-4-carbonitrile
Figure imgf000364_0001
HPLC-MS (Method C: m/z = 300 (M+1 ); Rt. = 3.79 min.
Example 960
4-(4-Bromoρhenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000364_0002
This compound is commercially available (MENAI).
Example 961
N-[4-(5-Cyano-1 H-[1 ,2(3]triazol-4-yl)-phenyl]-acetamide
Figure imgf000364_0003
This compound is commercially available (MENAI).
Example 962 (General procedure (O)) S^'-ChlorobiphenyM-yO-SH-ti^.Sltriazole^-carbonitrile 5 053070
364
Figure imgf000365_0001
HPLC-MS (Method C): m/z = 281 (M+1); Rt = 4.22 min.
The compounds in the following examples are commercially available and may be prepared using a similar methodoEogy:
Example 963
4-(4-Trifluoromethoxyphenyl)-1 H-[1 ,2.3]triazole-5-carbonitrile
Figure imgf000365_0002
Example 964
4-Benzo[1 ,3]dioxol-5-yl-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000365_0003
Example 965 -(3-Trif!uoromef:hylpheπyl)-1 H-[i ,2,3]triazole-5-carbonitrile
Figure imgf000366_0001
Example 966
4-Pyridin-3-yl-1 W-{1 ,2,3]triazo!e-5-carbonittile
Figure imgf000366_0002
Example 967 4-(2,6-Dichlorophenyl)-1W-[1.2I3JtriazoIe-5-carbonitrile
Figure imgf000366_0003
Example 968
4-Thiophen-2-yl-1 H-[1 ,2.3]triazole-5-carbonitri!e
Figure imgf000366_0004
Example 969
3,5-Dfmethylisoxazole-4-carboxylic acid 4-(5-cyaπo1W-[1,2,3}triazol-4-yl)pheny! ester
Figure imgf000367_0001
Example 970
3,3-Dimethyl-butyric acid 4-{5-cyano-1H-[1,2,3]triazol-4-yl)phenyl ester
Figure imgf000367_0002
Example 971
4-Methy!-[1,2,3]thiadiazole-5-carboxylic acid 4-(5-cyano-1H-[1,2,3]triazol-4-yl)phenyl ester
Figure imgf000367_0003
CH3
Example 972
4-Chlorobeπzoic acid 4-(5-cyano-1 H-[1 ,2,3]triazol-4-yl)phenyl ester
Figure imgf000367_0004
Example 973 4-(3-Phenoxyphenyl)-1 H-[λ ,2,3]triazole-5-carbonitrile
Figure imgf000368_0001
Example 974
4-(5-Bromo-2-methoxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile
Figure imgf000368_0002
Example 975
4-(2-Chtorα-6-fluoropheπyl)-1 H-[1 ,2,3]triazole-5-carbonitrj[e
Figure imgf000368_0003
The following cyanotriazoles are also compounds of the invention:
4-(2-Chloro-6-fluorophenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. Terephthalic acid monof 4-(5-cyano-1 W-[1,2,3]triazol-4-yl)phenyr| ester. N- [4-(5-cyano-1H-[1,2,3]triazol-4-yl)-phenyl]terephthalamic acid 4-(4-Octyloxyphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile 4-(4-Styrylphenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(4'-Trifluoromethytbiphenyl-4-yl)-1H~t1l2,3]triazole-5-carbonitril6. 4-(4'-Chlorobiphenyl-4-yl)-1 H-["\ ,2,3]triazole-5-carbonitrile. 4-(4'-Methoxybiphenyl-4-yl)-1 H-[A ,2,3]triazole-5-carbonitrile.
Figure imgf000368_0004
4-(9-Anthranyl)-1 H-[1 ,2.3]triazole-5-carbonitrile. 4-(4-Methoxy-1 -naphthyl)-1 H-[1 ,2,3]triazole-5-carbonitri!e. 4-(4-Aminophenyl)-1 H-[1 ,2,3]triazole-5-carbonitrile. 4-(2-Naphthyl)-1 H-[1 ,2,3]triazole-5-carbonitrile.
General procedure (P) for preparation of compounds of general formula li5:
Figure imgf000369_0001
wherein n is 1 or 3-20, AA is as defined above, R" is a standard carboxylic acid protecting group, such as Ci-Cθ-alkyl or benzyl and Lea is a leaving group, such as chloro, bromo, iodo, methanesulfonyloxy, toluenesulfonyloxy or the like.
This procedure is very similar to general procedure (D), steps 1 and 2 are identical.
Steps 3 and 4 are described in the literature (eg Beck & Gϋnther, Chem. Ber., 106, 2758-66 (1973))
Step 3 is a Knoevenagel condensation of the aldehyde obtained in step 2 with phenylsulfon- ylacetonitrile and step 4 is a reaction of the vinylsulfonyl compound obtained in step 3 with sodium azide. This reaction is usually performed in DMF at 90 — 1100C.
This General procedure (P) is further illustrated in the following two examples Example 976 (General procedure (P))
5-[6-(5-Cyano-1H-[1,2,3Itriazol-4-y[)-naphthalen-2-y[oxy]-pentanoic acid ethyl ester
Figure imgf000370_0001
6-Hydroxynaphthalene-2-carbaldehyde (Syncom BV. NL, 15.5 g, 90 mmol) and K2CO3 (62.2 g. 450 mmol) were mixed in DMF (30OmL) and stirred at room temperature for 1hour. Ethyl 5-bromovalerate (21.65 g, 103.5 mmol) was added and the mixture was stirred at room tem¬ perature for 16 hours. Activated carbon was added and the mixture was filtered. The filtrate was evaporated to dryness in vacuo to afford 28.4 g of crude 5-(6-formylnaphthalen-2- yloxy)pentanoic acid ethyl ester, which was used without further purification.
HPLC-MS (Method C ): m/z = 301 (M+1); Rt. = 4.39 min.
5-(6-Formylπaphthaleπ-2-ytoxy)peπtaπotc acid ethyl ester (28.4 g, 94.5 mmol), prteπylsulfon- ^ylacetonitrile (20.69, 113.5 mmol), and piperidine (0.94 mL) weretlissolved in DMF (200 mL) and the mixture was heated at 50 0C for 16 hours. The resulting mixture was evaporated to dryness in vacuo and the residue was dried for 16 hours at 40 0C in vacuo. The solid was recrystallised from 2-propanol (800 mL) and dried again as described above. This afforded 35 g (80%) of 5-[6-(2-benzenesulfonyl-2-cyanovinyl)naphthalen-2-yloxy]pentanoic acid ethyl ester.
HPLC-MS (Method C): m/z = 486 (M+23); Rt. = 5.09 min.
5-[6-(2-Ben2enesulfonyl-2-cyanovinyl)naphthalen-2-yloxy]pentanoic acid ethyl ester (35 g, 74.6 mmol) and sodium azide (4.9 g, 75,6 mmol) were dissolved in DMF (100 mL) and stirred for 16 hours at 500C- The mixture was evaporated to dryness in vacuo, redissolved in THF / ethanol and a small amount of precipitate was filtered off. The resulting filtrate was poured into water (2.5 L). Filtration afforded after drying 24.5 g (88%) of 5-[6-(5-cyano-1 H- [1,2,3Jtriazol-4-yl)naphthalen-2-yloxy]pentanoic acid ethyl ester (24.5 g, 88%). HPLC-MS (Method C): m/z = 365 (M+1); Rt. = 4.36 min.
Example 977 (General procedure (B))
5-[6-(5-Cyaπo-1 H-[1 ,2,3]triazσl-4-yI)-naphthalen-2-yloxy]-pentanoic acid
Figure imgf000371_0001
5-[6-(5-CyQnO-1 H-[1,2,3]triazol-4-yl)naphthaleπ-2-yloxy]peπtanoicacid ethyl ester (24.5 g, 67.4 mmol) was dissolved in THF (150 mL) and mixed with sodium hydroxide (8.1 g, 202 mmol) dissolved in water (50 mL). The mixture was stirred for 2 days and the volatiles were evaporated in vacuo. The resulting aqueous solution was poured into a mixture of water (1 L) and hydrochloric acid (1N, 250 mL}. The sofid was isolated by filtration, dissolved in sodium hydroxide (1 N, 200 mL), and the solution was washed with DCM and then ethyl acetate, the aquous layer was acidified with hydrochloric acid (12N). The precipitate was isolated by filtra¬ tion, dissolved in THF / diethyl ether, the solution was treated with MgSO4 and activated car¬ bon, filtrated and evaporated in vacuo to almost dryness followed by precipitation by addition of peπtane (1L). This afforded after drying in vacuo 17.2 g ( 76%) of the title compound.
HPLC-MS (Method C): m/z = 337 (M+1); Rt. = 3.49 min.
Example 978 (General procedure (P)) 6-[6-(5-Cyano-1 H-[1 ,2,3]triazol-4-yl)naphthalen-2-yloxy]hexanoic acid
Figure imgf000371_0002
HPLC-MS (Method C): m/z = 351 (M+1); Rt = 3.68 min.
Example 979 (General procedure (P)) 11-I6-(5-Cyano-1 H-[1 ^.SJtriazol^yl^naphthalen^-yloxyJ-undecanoic acid
Figure imgf000372_0001
HPLC-MS (Method C): m/z = 443 (tø+23); Rt =* 4.92 min.
Example 980 (General procedure (P))
2-{3-[6-(5-Cyaπo-1H-[1,2,3]triazoI-4-y|)-naphthalen-2-yloxy]-propyl}-rπaloriic acid diethyl ester
Figure imgf000372_0002
HPLC-MS (Method C): m/z = 465 (M+1); Rt. a 4.95 min.
Example 981 (General procedure (P)) a^δ-te-tS-Cyano-IH-fi^.sμriazoM-ylJ-naphthalen^-yfoxyJ-pentyl^malonic acid diethyl ester
Figure imgf000372_0003
HPLC-MS (Method C): m/z = 465 (M+1); Rt. = 4.95 min.
Example 982 (General procedure (P))
2-{3-[6-(5>Cyano-1 H-[1 ,2,3]triazol-4-y[)-naphthalen-2-yloxy]-propyl}-malonic acid
Figure imgf000373_0001
HPLC-MS (Method C): m/z = 381 (M+1); Rt. = 3.12 min.
Example 983 (General procedure (P)) 2-{5-[6-(5-Cyaπo-1 H-[1 ,2,3]triazol-4-yl)-naphthalen-2-yloxy]-pentyl}-malonic acid
Figure imgf000373_0002
HPLC-MS (Method C): m/z 0409 (M+1); Rt. = 3.51 min.
Example 984 (General procedure (P)) 4-[4-(5-Cyano-1 H-[1 ^.SltriazoW-ylJ-phenoxylbutyric acid
Figure imgf000373_0003
HPLC-MS (Method C): m/z = 273 (M+1); Rt = 2.44 min.
The following compounds may be prepared according to this general procedure (P):
4-(4-(5-Cyano-1H-[1 ,2,3]triazol~4-yl)phenoxy)butyric acid:
Figure imgf000374_0001
2-(4-(5-Cyaπo-1 W-[1 ,2,3]triazol-4-yl)phenoxy)acetic acid:
Figure imgf000374_0002
4-(4-(5-Cyano-1 H-Ji ,2,3]triazo1-4~yl)phenoxy)butyric acid ethyl ester 5-(4-(5-Cyano-1 H-[1 ,2,3]triazol-4~yl)phenoxy)pentanoic acid S-(4-(5-Cyaπo-1 W-[1,2,3]triazσ!-4-yl)phenoκy)octanoic acid 10-(4-(5-Cyano-1 W-[1 ,2,3]triazol-4-yl)phenoxy)decanoic acid 12-(4-(5-Cyano-1H-[i ,2,3]triazol-4-yl)phenoxy)clodecanoic acid
General procedure (R) for preparation,^ compounds of general formula 1i2:
/ DMF
Figure imgf000374_0003
- 8:1 :1
Figure imgf000374_0004
Figure imgf000374_0005
'12
Pol
Figure imgf000374_0006
wherein T is as defined above and R2 and R3 are hydrogen, aryl or lower alky!, both option¬ ally substituted.
The general procedure (R) is further illustrated by the following example:
Example 985 (General procedure (R)) Phenyl-[3-(2H-tetrazol-5-yl)-carbazo!-9-yl]-rnethanone
Figure imgf000375_0001
2-Chlorotritylchloride resin (100 mg, 0.114 mmol active chloride) was swelled in dichloro- methane (4 mL) for 30 minutes. The solvent was drained, and a solution of 3-(2H-tetrazol-5- yl)-9H-carbazole (80 mg, 0.34 mmol) in a mixture of N.N-dimethylformamide / dichloro- methane / N,N-di(2-propyl)ethylanrtine (5:5:1) (3 mL) was added. The reaction mixture was shaken at room temperature for 20 hours. The solvent was removed by filtration, and the resin was washed thoroughly with N,N-dimethylformarnide (2 x 4 mL) and dichloromethaπe (6 x 4 mL). A solution of 4-(dimethylamino)pyridine (14 mg, 0.11 mmol) and N,N-di(2- propyl)etnylamine (0.23 mL, 171 mg, 1.32 mmol) in N,N-dimethylformamide (2 mL) was added followed by benzoyl chloride (0.13 mL, 157 mg, 1.12 mmol). The mixture was shaken for 48 hours at room temperature. The drained resin was washed consecutively with di- chloromethane (2 x 4 mL), methanol (2 x 4 mL) and tetrahydrofuran (4 mL). The resin was treated for 2 hours at room temperature with a solution of dry hydrogen chloride in tetrahy¬ drofuran / ethyl ether / ethanol = 8:1:1 (0.1 M, 3 mL). The reaction mixture was drained and concentrated. The crude product was stripped with dichloromethane (1.5 mL) three times to yield the title compound.
HPLC-MS (Method C): m/z: 340 (M+1); Rt = 3.68 min.
1H-NMR (DMSO-dβ): δ 8.91 (1H, s). 8.34 (1H, d). 8.05 (1H. d), 7.78 (3H, m), 7.63 (3H, m),
7.46 (2H. m), 7.33 (1 H, dd).
The compounds in the following examples were prepared in a similar fashion. Example 986 (General procedure (R)) Phenyl-[5-(2H-tetrazol-5-yI)-indol-1-yl]-metrιanone
Figure imgf000376_0001
HPLC-MS (Method C): m/z: 290 (M+1); Rt = 3.04 min.
1H-NMR (DMSO-dβ): δ 8.46 (1H. d), 8.42 (1H, d), 8.08 (1H1 dd), 7.82 (2H, d), 7.74 (1H, t),
7.64 (2H, 0, 7.55 (1H1 d), 6.93 (1H, d).
Example 987 (General procedure (R)) (2,3-Difluorophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-ylJ-methanone
Figure imgf000376_0002
hPLC-MS (Method B): m/z - 326 (M+1); Rt = 3.85 min.
Example 988 (General procedure (R)) (2-Fluoro-3-tvifluoromethylpheny<H5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000376_0003
HPLC-MS (Method B): m/z = 376 (M+1); Rt = 4.32 min.
Example 989 (General procedure (R)) (3-Nitrophenyl)-[5-(2H-tetrazol-5-yl)-indol-1-yl]-methanone
Figure imgf000377_0001
HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.72 min.
Example 990 {General procedure (R)) (4-Nitrophenyl)~[5-(2H4etrazol-5-yI)-indol-1-yl]-methanone
Figure imgf000377_0002
HPLC-MS (Method B): m/z = 335 (M+1); Rt = 3.71 min.
Example 991 (General procedure (R)) Naphthalen-2-yl-[5-(2H-tetrazol-5-yl)-indol-1 -yl]-methanone
Figure imgf000377_0003
HPLC-MS (Method C): m/z = 340 (M+1); Rt = 4.25 min.
Example 992 (General procedure (R))
Figure imgf000377_0004
HPLC-MS (Method C): m/z: 354 (M+1); Rt = 3.91 min. Example 993 (General procedure (R))
Figure imgf000378_0001
HPLC-MS (Method C): m/z: 418 (M+ 1); Rt = 4.39 min.
Example 994 (General procedure (R))
Figure imgf000378_0002
HPLC-MS (Method C): m/z: 370 (M+1); Rt = 4.01 min.
Example 995 (General procedure (R))
Figure imgf000378_0003
HPLC-MS (Method C): m/z: 374 (M+1); Rt = 4.28 min.
Example 996 (General procedure (R))
Figure imgf000378_0004
HPLC-MS (Method C): m/z: 416 (M+1); Rt = 4.55 min.
Example 997 (General procedure (R))
Figure imgf000379_0001
HPLC-MS (Method C): m/z: 354 (M+1 ); Rt = 4.22 min.
Example 998 (General procedure (R))
Figure imgf000379_0002
HPLC-MS (Method C): m/z: 358 (M+1 ); Rt = 3.91 min.
Example 999 (General procedure (R))
Figure imgf000379_0003
HPLC-MS (Method C): m/z: 390 (M+1); Rt = 4.38 min.
Example 1000 (General procedure (R))
Figure imgf000379_0004
HPLC-MS (Method C): m/z: 418 (M+1); Rt = 4.36 min.
Example 1001 (General procedure (R))
Figure imgf000380_0001
HPLC-MS (Method C): m/z: 304 (M+1 ); Rt = 3.32 min.
Example 1002 (General procedure (R))
Figure imgf000380_0002
HRLC-MS (Method C): m/z: 368 (M+1 ); Rt = 3.84 min.
Example 1003 (General procedure (R))
Figure imgf000380_0003
HPLC-MS (Method C): m/z: 320 (M+1); Rt = 3.44 min. Example 1004 (General procedure (R))
Figure imgf000381_0001
HPLC-MS (Method C): m/z: 324 (M+ 1); Rt = 3.73 min.
Example 1005 (General procedure (R))
Figure imgf000381_0002
HPLC-MS (Method C): m/z: 304 (M+1); Rt = 3.64 min.
Example 1006 (General procedure (R))
Figure imgf000381_0003
HPLC-MS (Method A): m/z: 30θ (M+1); Rt = 3.61 min.
Example 1007 (General procedure (R))
Figure imgf000382_0001
HPLC-MS (Method C): m/z: 368 (M+1); Rt = 3.77 min.
Example 1008 (General procedure (R))
Figure imgf000382_0002
HPLC-MS (Method A): (sciex) m/z: 326 (M+1); Rt = 3.73 min. HPLC-MS (Method C): m/z: 326 (M+1); Rt = 3.37 min.
Example 1009 (General procedure (R))
Figure imgf000382_0003
HPLC-MS (Method C): m/z: 374 (M+1); Rt = 4.03 min.
General procedure (Q) for preparation of compounds of general formula li7:
Step 1 Step 2 CGr- Lnk-Frg-CO2H ■" CGr- Lπk-Fig-CO-Lea *- CGr-Lnk-Frg-CO-Protamine
I17 wherein CGr, Lnk and Frg is as described above; and Lea is a suitable leaving group, such as N- hydroxysuccinimide, 1-hydroxybenzotriazole, benzotriazole, N-hydroxy-5-norbornene-endo- 2,3-dicarboximide.
Step 1 , activation of a carboxylic acid:
Step 1 is well known to those skilled in the art, see e.g. F. Z. Dόrwald, "Organic Synthesis on Solid Phase", Witey-VCH 2000, chapter 13, and refeences cited therein. The CGr-Lnk-Frg- CO-Lea is prepared using the carboxylic acid and H-Lea using a condensation reagent, such as N.Λf-dicyclohexylcarbondiimide, diisopropylcarbodiimide, carbonyldiimidazole, 1-ethyl-3- (3'-dirnethylaminopropyl)carbodiimide (hydrochloride), 2-ethoχy-1 -ethoxycarbonyl-1 ,2-di- hydroquinoline or the like. Alternatively CGr-Lnk-Frg-CO-Lea is be prepared using the car¬ boxylic acid and a reagent that combines delivery of Lea and mediates condensation, such as 2^1H-benzotriazole-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), 2- (1H-benzotriazole-1-yl)-1 ,1 ,3,3-tetramethyIuroniυm tertafluoroborate (TBTU), N,W-disuccin- imidyl carbonate, benzotriazole-1-yloxy-tris-pyrrolidinophosphonium hexafluorophosphate (PyBOP)1 benzotriazote-1 -yloxy-tris-dimethylaminophosphonium hexafluorophosphate (BOP), or the like.
Step 2, acylation of protamine with the activated carboxylic acid obtained in step 1 : The reaction medium is water, optionally using organic co-solvents, such as N,N-dimethyl- formamide, N-methylpyrrolidinone, acetonitrile, dimethyl sulfoxide. A base, such as triethyl amine, potassium or sodium carbonate, potassium or sodium hydroxide, can optionally be used. When protamine sulphate is used, prior to acylation, the sulphate salt is converted to a more soluble salt, such as the acetate, using e.g. barium acetate.
Optionally, the zink-binding group CGr is attached via an adequate linker to the C-terminal of the protamine. This may be done by coupling a mono-Boc protected diamine to a carboxylic acid on the CGr, followed by deprotection, for example as described in example 492 to ob¬ tain a CGr carrying an amino group. Other CGr-carboxylic acids includes, but is not limited to, those of examples 738, 743, 283, 466, 372, 460, and 467.
Protamine may be coupled C-terrninally to the CGr carrying an amino group by means of, for example, activation to the N-hydroxysucclnimide ester. Optionally, the N-terminal of the pro¬ tamine is temporarily protected, for example by a Boc group.
This general procedure (Q) is further illustrated in the following examples 1010 and 1011. Preparation of starting materials:
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethy])naphtha1en-1-yloxy]butyric acid 1 -hydroxy- sυccinimidy! ester
Figure imgf000384_0001
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-ylO5cy]butyric acid (1 g, 2.79 mmol, prepared as described in WO 200327081. example 291 ) was dissolved in THF (10 ml) and DMF (5 ml). 0-(Λ/-Succinimidyl)-1,1,3,3-tetramethyluronium tetrafluoroborate (TSTU, 1.01 g, 3.36 mmol) and N,N-diisopropylethylamine (DIPEA, 0.574 ml, 3.36 mmol) were added. The resulting mixture was stirred at room temperature for 3 days and 1 N hydrochloric acid (200 ml) was added. The mixture was extracted with ethyl acetate (150 ml). The organic extract was washed with water (3 x 150 ml), dried (Na2SO4) and concentrated in vacuo. This af¬ forded 1.13 g (89%) of the title compound.
1H NMR (DMSO-c/β): δ = 2.26 (2H, p), 2.82 (4H, s), 3.00 (2H, t).4.34 (2H, t), 7.15 (1H, d), 7.61-7.73 (3H, m), 8.12 (1H, d), 8.32 <1H, d), 8.38 (1H, s), 12.6 (1H, bs).
The following intermediates were similarly prepared:
4-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoic acid 1-hydroxysuccinimidyl ester
Figure imgf000384_0002
1H NMR (DMSO-dβ): δ - 2.86 (4H, s), 5.93 (2H, s). 7.33 (1H, t), 7.40 (2H, d), 7.52 (1H1 1), 7.67 (1H, d), 7.87 (1H, d), 8.03 (2H, d), 8.11 (1H, dd), 8.31 (1H, d), 8.91 (1H. d). 5-[6-(5-Cyaπo-1 H-[\ ,2,3]triazol-4-yl)ngphthalen-2-yloxy]pentaπoic acid 1-hydroxysuccinimidyl ester
Figure imgf000385_0001
1H NMR (DMSO-Cf6): δ = 1.8 - 1.9 (4H, m), 2.81 (2H, t).2.83 (4H, s), 4.18 (2H, t), 7.29 (1H, dd), 7.44 (1H, d), 7.91 (1H, d), 7.96 (1H, d), 8.02 (1H, d), 8.38 (1H, s).
Example 1010 (General Procedure (Q)
4-[4-(2,4-Dioxothiazolidin-5-ylideπemethyl)naphthalen-1 -yloxy]butyrylprotamlne, acetate salt Major components: R R R R R S S S R P I R R R R R P R A S R R R R R G G R R R R—w
R R R R S S R R P V R R R R Ft P R V S R R R R R R G G R R R R—en
R R R R S S S R P V R R R R R P R V S R R R R R R G G R R R R—OM
R R R R A S R R I R R R R R P R V S R R R R R G G R R R R
Figure imgf000385_0002
3.22 g salmon protamine sulphate was added water (200 ml) and heated to 450C until com¬ plete dissolution and cooled to room temperature. Barium acetate {1.6 g) dissolved in water (30 ml) was added and left at room temperature for 1 hour. The mixture was filtered and the filtrate was centrifuged. The clear supernatant was made basic (pH 10.8) by addition of an aqueous solution of triethyl amine.4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)πaphthalen-1- yloxyjbutyric acid 2,5-dioxopyrrolidin-1-yl ester (0.49 g) dissolved in NMP (200 ml) was added (optionally with cooling) and the mixture was left at room temperature for 16 hours. The product was precipitated by addition of 2 N sulphuric acid (to pH 2.7) followed by addi¬ tion of 2-propanol (1000 ml). The supernatant was decanted and the residue was washed twice with 2-propanol using centrifugation and decantation. The solid was dissolved in warm (70 "C) water (130 ml) and barium acetate (7.5 g) was added. The mixture was left at room temperature 2 hours. The mixture was added 2-propanol (1000 ml) and left at room tempera¬ ture for 16 hours. The mixture was filtered and the filtrate was concentrated in vacuo. This afforded 3.1 g crude product. This was dissolved in ultrapure water (25 ml) and desalted on 10 NAP 25 columns (Sephadex G25 DNA grade, Amersham) by addition of 2.5 ml to each column. The columns were each eluted with 3.5 ml ultrapure water. The combined eluates were lyophilised to afford 450 mg of the title material.
MALDI-TOF: m/z = 4403, 4577, 4589, and 4659 (selected peaks}.
Example 1011 (General Procedure (Q))
{5-[6-{5-Cyano-1H-[1 ,2,3]triazαl-4-yl)naphthalen-2-yloxy]pentyl}protamine, acetate salt Major components:
β R R R R-
G R R R —«
G R R R R—W
Figure imgf000386_0001
4.0 g salmon protamine sulphate was added water (200 ml) and heated to 55 "C until com¬ plete dissolution and cooled to room temperature. Barium acetate (2 g) dissolved in water (25 ml) was added and left at room temperature for 1.5 hour. The mixture was centrifuged. The clear supernatant was made basic (pH 10.45) by addition of an aqueous mixture of triethyl amine. S-Iδ-CS-Cyano-iH-II^.SltriazoM-yO-naphthalen^-yloxyJ-pentanoic acid 2,5- dioxopyrrolidin-1-yl ester (0.65 g) dissolved in NMP (200 ml) was added slowly (with cooling, the temperature was kept below 15 "C) and the mixture was left at room temperature for 16 hours. The product was precipitated by addition of 2 N sulphuric acid (to pH 2.5) followed by addition of 2-propanol (1500 ml). The supernatant was decanted and the residue was washed twice with 2-propanol using centrifugation and decantation. The solid was dissolved in warm (70 "C) water (175 ml) and barium acetate (2 g) was added. The mixture was left at room temperature for 16 hours. The mixture was centrifuged and the supernatant was lyophi¬ lised. This afforded 4.55 g crude product. 2.23 g of this was dissolved in water (100 ml) and added 25% ammonia (to pH 11 , approx. 20 ml) and the mixture was left at room temperature for 16 hours. Acetic acid glacial was added to pH 3.7 and the mixture was lyophilised. This afforded 20 g. 12 g of this was dissolved in 0.5M acetic acid (32 ml) and de-salted in two runs on two Sephadex G25 fine, HiPrep 26/10 columns (26 mm ID, L = 100 mm, connected in series) elυting 2 ml/miπ with 0.5M acetic acid. Salt-free fractions containing the desired compound were lyophilised to afford 610 mg of the title material.
MALDI-TOF: m/z = 4385, 4557, 4571, and 4642 (selected peaks).
Other compounds of the present invention include:
Building block from example 470:
5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)naphthalen-1-yloxy]pentanoyl-protamine
Building block from example 283:
4-[4-(2,4-Dioxothiazo1idiπ-5-ylmethyl)naphthalen-1-yloxy]butyryl-protamine
Building block from example 476:
2-[4-(2,4-Dioxothiazolidin-5-ylidenemethy1)naphthalen-1-y1oxy]acetyl-protamine
Building block from example 480:
2-{5-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)benzy1idene]-4-oxo-2-thioxothiazolidin-3- yl}acetyl-protamine
2-{5-[4-(2,4-Dioxothiazolidin-5-ylmethyl)benzyl]-4-oxo-2-thioxothiazolidin-3-yl}acetyl- protamine
Building block from example 462:
3-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenyl]acryloyl-protamine
Building block from example 473:
2-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-protarnine
Building block from example 466:
4-[2-Chloro-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-protamine
Building block from example 460:
4-[4-(2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-protamine
Building block from example 467:
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylidenemethyl)phenoxy]butyryt-protamine
Building block from example 464:
4-(2,4-Dioxothiazolidln-5-ylidenemethyl)ben2oyl-protamine
[3-(2,4-Dioxothiazo1idin-5-ylidenemethyl)indol-1-yl]acetyl-protamine
3-[3-(2,4-Dioxothiazolidin-5-ylidenemethyl)indol-1-yl]propionyl-protamine
4-[5-Bromo-6-(2,4-dioxothiazolidin-5-ylidenemethyl)-naphthalen-2-yloxymethyl]-benzoy1- protamine 4-(2,4-Dioxothiazolidin-5-ylmethyl)benzoyJ-protamine
[3-(2,4-Dioxothiazolidin-5-ylmethyl)indol-1-yl]acetyl-protamine
3-[3-(2,4-Dioxothiazolidin-5-ylmethyl)indol-1-yl]propionyl-protamine
4-[5-Bromo-6-(2r4-dioxothiazolidin-5-ylmethyl)naphthalen-2-yloxymethyl]benzoyl-protamine
Building block from example 463:
2-[4-(2I4-Dioxothiazolidin-5-ylidenβmethyl)phenoxy]acetyl-protamine
2-[4-(2,4-Dioxothiazolidin-5-ylmethyl)phenoxy]acetyl-protamiπe
Building block from example 461:
2-[3-{2,4-Dioxothiazolidin-5-ylidenemethyl)phenoxy]acetyl-protamine
2-[3-(2,4-Dioxothiazolidin-5-ylmethyl)phenoxy]acetyl-protamine
Building block from example 474:
4-I3-(2,4-Dioxothiazolidin-5-yltdenemethyl)phenoxy]butyryl-protaπnine
4-[3-(2,4-Dioxothiazolidin-5-y1methyl)phenoxy]butyryl-protamine
Building block from example 468:
4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylidenemethyl)phenoxy]butyryl-protamine
4-[2-Bromo-4-(4-oxo-2-thioxothiazolidin-5-ylmethyl)phenoxy]butyryl-protamine
Building block from example 359 :
5-[4-(2l4-Dioxothiazolidin-5-ylidenemethyi)naphthalen-1-yloxy]-peπtanoyl-protamine
5-[4-{2,4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]-pentanoyl-protamine
Building block from example 360 :
4-[3-(2,4-Dioxothiazolidin-5-ylidenerπethyl)-1,4-dimethylcarbazol-9-ylnπethyl]-benzoyl- protamine
4-t3-(2,4-Dioxothiazolidin-5-ytmethyl)-1,4-diinethyIcarbazol-9-ylmethyl]-benzoyl-protaπnine
Building block from example 738:
4-[3-{2H-Tetrazol-5-yl)carbazol-9-ylmethyl]benzoyl-protamiπe
Building block from page 319:
4'-[3-(2H-Tetrazol-5-yl)carbazol-9-ylmethyl]biphenyl-4-carboπyl-protamine
Building block from example 743:
3-[3-(2H-Tetrazot-5-yl)carbazol-9-ylmethyl]benzoy!-protamine
Building block from example 809 :
4'-[5-(2H-Tetrazol-5-yl)-indol-1-ylmethyl]-biphenyl-4-carbonyl-protamine
Reduced building block of example 476 :
[4-(2l4-Dioxothiazolidin-5-ylmethyl)naphthalen-1-yloxy]acetyl-protamiπe
Reduced building block of example 467 :
4-[2-Bromo-4-(2,4-dioxothiazolidin-5-ylmethyl)phenoxy]butyry!-prolamine
Reduced building block of example 470 : 5-[4-(2,4-Dioxothiazolidin-5-ylmethyl)naphthaleπ-1-yloxy]pentanoyl-protamine Reduced building block of example 462 : 3-[4-(2,4-Dioxothiazolidin-5-ylmethyI)phenyl]acryloyl-protamine Reduced building block of example 466 :
4-t2-Chloro-4-(2,4-dtoxothiazolidin-5-ylmethyl)phenoxy]butyryl-protamine Reduced building block of example 473 :
[2-Bromo-4-{2,4-dioxothiazolidin-5-ylmethyl)phenoxy]acetyl-protamine Reduced building block of example 460 : 4-[4-(2,4-Dioxothiazolidin-5-ylmethyl)phenoxy]butyryl-protamine
Example 1012
The PD studies were performed on domestic female pigs, LYD cross-breed, weigh¬ ing between 55 and 110 kg. The pigs were catheterised into the jugular vein through an ear vein at least 2 days before start of the study. The last meal before the start of the study was served to the animals approx. 18 hours prior to the injection of the test preparation, and the animals had free access to water at all time during the fasting period and the test period.
At time 0 hours the test preparation was given subcutaneous on the lateral side of the neck. At regular time intervals blood samples were drawn from the cathether and sam¬ pled into 1.5 ml glass tubes pre-coated with heparin. The blood samples were kept in ice wa- ter until separation of plasma by centrifugation for 10 min 3000 rpm at 4°C, which was done within the first 30 minutes. Plasma samples were stored at 4°C for short time (2-3 hours) or at -18βC for long term storage and were analysed for glucose on COBAS MIRA. Estimation of start level of glucose in blood was done on 4 samples taken 1 hour, half an hour, 20'min- utes and immediately before injection. Figure 1. Change in plasma glucose level (% of baseline) (average +/- SEM of 3 pigs) after subcutaneous injection at 0 hours of the following preparation 0.6mM A21G, B28D insulin, 0.3mM Zn2+, 3OmM phenol, 1.6% glycerol, 0.3mM 4-[4-(2,4-Dioxothiazolidin-5- y!idenemethyl)naphthalen-1-yloxy]butyrylprotamine (Example 1010), pH=3.0.
ANALYTICAL METHODS Assays to quantify the binding affinity of ligands to the metal site of the insulin R6 hexamers:
4H3N-assay:
The binding affinity of ligands to the metal site of insulin R6 hexamers are measured in a UV/vis based displacement assay. The UV/vis spectrum of 3-hydroxy-4-nitro benzoic acid (4H3N) which is a known ligand for the metal site of insulin R6 shows a shift in absorption maximum upon displacement from the metal site to the solution (Huang et a!.. 1997, Bio¬ chemistry 36, 9878-9888). Titration of a ligand to a solution of insulin R6 hexamers with 4H3N mounted in the metal site allows the binding affinity of these ligands to be determined follow¬ ing the reduction of absorption at 444 πm.
A stock solution with the following composition 0.2 mM human insulin, 0.067 mM Zn-acetate, 40 mM phenol, 0.101 mM 4H3N is prepared in a 1OmL quantum as described below. Buffer is always 5OmM tris buffer adjusted to pH=8.0 with NaOH/CICV -
1000 μL of 2.OmM human insulin in buffer 66.7 μL of 1OmM Zn-acetate in buffer 800 μL of 50OmM phenol in H2O 201 μL θf 4H3N in H2O 7.93 ml buffer
The ligand is dissolved in DMSO to a concentration of 20 mM.
The ligand solution is titrated to a cuvette containing 2 mL stock solution and after each addi¬ tion the UV/vis spectrum is measured. The titration points are listed in Table 1 below.
Table 1
Figure imgf000390_0001
The UV/vis spectra resulting from a titration of the compound 3-hydroxy-2-naphthoic acid is shown in Figure 2. Inserted in the upper right corner is the absorbance at 444nm vs. the con- centration of ligand. The following equation is fitted to these datapoints to determine the two parameters KD(obs), the observed dissociation constant, and abSmax the absorbance at maximal ligand concentra¬ tion.
abs ([ligand],™) = (absmax • [ligand]^)/ (KD(obs) + [ligandW)
The observed dissociation constant is recalculated to obtain the apparent dissociation con¬ stant
Ko(app) = K0(ObS) / ( 1+[4H3N]/K4H3N )
The value of
Figure imgf000391_0001
is taken from Huang et al., 1997, Biochemistry 36, 9878-9888.
TZD-assay:
The binding affinity of ligands to the metal site of insulin R6 hexamers are measured in a fluo- rescense based displacement assay. The fluorescence of 5-(4-dimethylaminobenzylidene)- thiazolidine-2,4-dione (TZD) which is a ligand for the metal site of insulin R6 is quenched upon displacement from the metal site to the solution. Titration of a ligand to a stock solution of insulin R6 hexamers with this compound mounted in the metal site allows the binding affin¬ ity of these ligands to be determined measuring the fluorescence at 455πm upon excitation at 4i0nm.
Preparation
Stock solution: 0.02 mM human insulin, 0.007 mM Zn-acetate, 40 mM phenol, 0.01 mM TZD in 5OmM tris buffer adjusted to pH=8.0 with NaOHZCIO4 '. The ligand is dissolved in DMSO to a concentration of 5 mM and added in aliquots to the stock solution to final concentrations of 0-250 μM.
Measurements
Fluorescence measurements were carried out on a Perkin Elmer Spectrofluorometer LS50B.The main absorption band was excited at 410 nm and emission was detected at 455 nm. The resolution was 10 nm and 2.5 nm for excitation and emission, respectively.
The fluorescence spectra resulting from a titration of the compound 5-(4-dimethylamino- benzylidene)thiazolidine-2,4-dione (TZD) is shown in Figure 3. Inserted in the upper right corner is the fluorescence at 455 nm upon exitation at 410 nM vs. the concentration of ligand.
Data analysis This equation is fitted to the datapoints
ΔF(455nm)) = ΔFmax • [ligandWt K^app) • ( 1+rrZD]/Kraj)+ [ligand]^))
KD(app) is the apparent dissociation constant and Fmax is the fluorescence at maximal ligand concentration. The value of KTZD is measured separately to 230 nM
Two different fitting-procedures can be used. One in which both parameters, KD(app) and Frnax. are adjusted to best fit the data and a second in which the value of Fmax is fixed
Figure imgf000392_0001
and only Ko(app) is adjusted. The given data are from the second fitting procedure. The Solver module of Microsoft Excel can be used to generate the fits from the datapoints.

Claims

1. A pharmaceutical preparation comprising
• Insulin
• Zinc ions • A ligand which binds reversibly to a HisB10 Zn2+ site of an R-state hexamer and wherein the ligand is extended by covalent attachment to protamine, having the following general formula (I)
CGr-Lnk-Frg-Protamine (I)
wherein:
CGr is a chemical group which reversibly binds to a His610 Zn2+ site of an insulin hexamer;
Lnk is a linker selected from
• a valence bond
• a chemical group GB of the formula -B1-B2-C(O)-, -B1-B2-SO2-, -B1-B2-CH2-, or -B1- B2-NH-; wherein B1 is a valence bond, -O-, -S-, or -NR6B-,
B2 is a valence bond, Ci-Ci8-alkylene, C2-Ci8-alkenylene, C2-Ci8-alkynylene, arylene, heteroarylene, -d-Cis-alkyl-aryl-, -C2-Ci 8-alkenyl-aryl-, -C2-C18-alkynyl-aryl-; -C(=O)-
Ci-C18-alkyl-C(=O)-, -C(=O)-Ci-C18-alkenyl-C(=O)-,
Figure imgf000393_0001
alkyl-C(=O)-, -C(=O)- C1-Ci8-alkyl-S-Ci-C18-alkyl-C(=O)-, -C(=O)-CrC18-alkyl-NR6-Cr Ci8-alkyl-C(=O)-, -C(=O)-aryl-C(=O)-, -C(=0)-heteroaryl-C(=0)-; wherein the alkylene, alkenylene, and alkynylene moieties are optionally substituted by -CN, -CF3, -OCF3, -OR6B, or -NR6BR7B and the arylene and heteroarylene moieties are optionally substituted by halogen, -C(O)OR68, -C(O)H, OCOR6B, -SO2, -CN, -CF3, -OCF3, -NO2, -0R6B, -NR6BR7B, CrCi8-alkyl, or CrCi8-alkanoyl; R6Band R7B are independently H, CrC4-alkyl;
Frg is a fragment consisting of O to 5 neutral α- or β-amino acids, or
a salt thereof with a pharmaceutically acceptable acid or base, or any optical isomer or mix¬ ture of optical isomers, including a racemic mixture, or any tautomeric forms.
2. A pharmaceutical preparation according to claim 1 wherein CGr is a chemical structure selected from the group consisting of carboxylates, dithiocarboxylates, phenolates, thiophe- nolates, alkylthiolates, sulfonamides, imidazoles, triazoles, 4-cyano-1 ,2,3-triazoles, benzimi- dazoles, benzotriazoles, purines, thiazolidinediones, tetrazoles, 5-mercaptotetra∑oles, rho- danines, N-hydroxyazoles, hydantoines, thiohydantoines, barbiturates, naphthoic acids and salicylic acids.
3. A pharmaceutical preparation according to claim 2 wherein CGr is a chemical structure selected from the group consisting of benzotriazoles, 3-hydroxy 2-napthoic acids, salicylic acids, tetrazoles, thiazolidinediones, 5-mercaptotetrazoles, or 4-cyano-1 ,2,3-triazoles.
4. A pharmaceutical composition according to any one of the claims 1 to 3 wherein CGr is
\ / IYT N=N
wherein K is a valence bond, CrC6-alkylene, -NH-C(=O)-U-, -Ci-Ce-alkyl-S-, -CrCe-alkyl-O-, -C(=O)-, or -C(=O)-NH-, wherein any CrC6-alkyl moiety is optionally substituted with R38,
U is a valence bond, Ci-C6-alkenylene, -Ci-C6-alkyl-O- or Ci-C6-alkylene wherein any C1- C6-alkyl moiety is optionally substituted with Ci-C6-alkyl,
R38 is Ci-C6-alkyl, aryl, wherein the alkyl or aryl moieties are optionally substituted with one or more substituents independently selected from R39,
R39 is independently selected from halogen, cyano, nitro, amino,
M is a valence bond, arylene or heteroarylene, wherein the aryl or heteroaryl moieties are optionally substituted with one or more substituents independently selected from R40,
R40 is selected from
• hydrogen, halogen, -CN1 -CH2CN, -CHF2, -CF3, -OCF3, -OCHF2, -OCH2CF3, -OCF2CHF2, -S(O)2CF3, -OS(O)2CF3, -SCF3, -NO2, -OR41, -NR41R42, -SR41, -NR41S(O)2R42, -S(O)2NR41R42, -S(O)NR41R42, -S(O)R41, -S(O)2R41, -OS(O)2 R41,
-C(O)NR41R42, -OC(O)NR41R42, -NR41C(O)R42, -CH2C(O)NR41R42, -OC1-C6- alkyl-C(O)NR41R42, -CH2OR41, -CH2OC(O)R41, -CH2NR41R42, -OC(O)R41, -OC1-C6- alkyl-C(O)OR4\ -OCrC6-alkyl-OR41, -S-CrC6-alkyl-C(O)OR4\ -C2-C6-alkenyl- C(=O)OR41, -NR41-C(=O)-CrC6-alkyl-C(=O)OR4\ -NR41-C(=O)-CrC6- alkenyl-C(=O)OR41 , -C(O)OR41, -C2-C6-alkenyl-C(=O)R41, =0, -NH-C(=O)-O-d- Ce-alkyl, or -N H-CC=O)-CC=O)-O-C1 -G6-alkyl,
• Ci-C6-alkyl, C2-C6-alkenyl or C2-C6-alkynyl, which may each optionally be substituted with one or more substituents selected from R43,
• aryl, aryloxy, aryloxycarbonyl, aroyl, arylsulfanyl, aryl-CrC6-alkoxy, aryl-CrC6-alkyl, aryl-C2-C6-alkenyl, aroyl-C2-C6-alkenyl, aryl-C2-C6-alkynyl, heteroaryl, heteroaryl-d- C6-alkyl, heteroaryl-C2-C6-alkenyl or heteroaryl-C2-C6-alkynyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from
R44,
R41 and R42 are independently selected from hydrogen, -OH, CrC6-alkyl, CrC6-alkenyl, aryl- d-Ce-alkyl or aryl, wherein the alkyl moieties may optionally be substituted with one or more substituents independently selected from R45, and the aryl moieties may optionally be substi¬ tuted with one or more substituents independently selected from R46; R41 and R42 when at¬ tached to the same nitrogen atom may form a 3 to 8 membered heterocyclic ring with the said nitrogen atom, the heterocyclic ring optionally containing one or two further heteroatoms selected from nitrogen, oxygen and sulphur, and optionally containing one or two double bonds,
R43 is independently selected from halogen, -CN1 -CF3, -OCF3, -OR41, and -NR41R42
R44 is independently selected from halogen, -C(O)OR41, -CH2C(O)OR41, -CH2OR41, -CN, - CF3, -OCF3, -NO2, -OR41, -NR41R42 and CrC6-alkyl,
R45 is independently selected from halogen, -CN, -CF3, -OCF3, -O-CrC6-alkyl, -C(O)-O-C1-
Ce-alkyl, -COOH and -NH2,
R46 is independently selected from halogen, -C(O)OC1 -C6-alkyl, -COOH, -CN, -CF3, -OCF3, -
NO2, -OH, -OCrC6-alkyl, -NH2, C(=0) or CrC6-alkyl,
Q is a valence bond, d-C6-alkylene, -d-Ce-alkyl-O-, -d-Ce-alkyl-NH-, -NH-d-C6-alkyl,
-NH-C(=O)-, -C(=O)-NH-, -O-d-Ce-alkyl, -C(=0)-, or -CrC6-alkyl-C(=O)-N(R47)- wherein the alkyl moieties are optionally substituted with one or more substituents independently selected from R48, R47 and R48 are independently selected from hydrogen, d-C6-alkyl, aryl optionally substituted with one or more R49,
R49 is independently selected from halogen and -COOH,
T is
• hydrogen,
• d-Ce-alkyl, C2-C6-alkenyl , C2-C6-alkynyl, d-Ce-alkyloxy-carbonyl, wherein the alkyl, alkenyl and alkynyl moieties are optionally substituted with one or more substituents independently selected from R50,
• aryl, aryloxy, aryloxy-carbonyl, aryl-CrC6-alkyl, aroyl, aryl-CrC6-alkoxy, aryl-C2- C6-alkenyl, aryl-C2-C6-alkyny-, heteroaryl, heteroaryl-CrC6-alkyl, heteroaryl-C2- C6-alkenyl, heteroaryl-C2-C6-alkynyl,
wherein any alkyl, alkenyl , alkynyl, aryl and heteroaryl moiety is optionally substituted with one or more substituents independently selected from R50,
R50 is d-Ce-alkyl, d-C6-alkoxy, aryl, aryloxy, aryl-Ci-C6-alkoxy, -C(=O)-NH-Ci-C6-alkyl-aryl, -C(=O)-NR50A-CrC6-alkyl, -C(=O)-NH-(CH2CH2O)mCrC6-alkyl-COOH, heteroaryl, het- eroaryl-d-Ce-alko-xy, -CrC6-alkyl-COOH, -O-Ci-Ce-alkyl-COOH, -S(O)2R51, -C2-C6-alkenyl- COOH, -OR51, -NO2, halogen, -COOH, -CF3, -CN, =0, -N(R51R52), wherein m is 1 , 2, 3 or 4, and wherein the aryl or heteroaryl moieties are optionally substituted with one or more R53, and the alkyl moieties are optionally substituted with one or more R50B. R50A and R50B are independently selected from -C(O)Od-C6-alkyl, -COOH, -CrC6-alkyl- C(O)OCi -Cβ-alkyl, -d-Cβ-alkyl-COOH, or CrC6-alkyl, R51 and R52 are independently selected from hydrogen and Ci-Ce-alkyl, R53 is independently selected from Ci-Ce-alkyl, d-C6-alkoxy, -d-Ce-alkyl-COOH, -C2- Ce-alkenyl-COOH, -OR51, -NO2, halogen, -COOH, -CF3, -CN, or -N(R51R52),
or any enantiomer, diastereomer, including a racemic mixture, tautomer as well as a salt thereof with a pharmaceutically acceptable acid or base.
5. A pharmaceutical composition according to claim 4 wherein K is a valence bond, d- Ce-alkylene, -NH-C(=O)-U-, -d-Ce-alkyl-S-, or -CrC6-alkyl-O, wherein any d-C6-alkyl moiety is optionally substituted with R38.
6. A pharmaceutical composition according to claim 4 or 5 wherein U is a valence bond or -d-Ce-alkyl-O-.
7. A pharmaceutical composition according to any one of the claims 4 to 6 wherein M is ary- lene or heteroarylene, wherein the arylene or heteroarylene moieties are optionally substi¬ tuted with one or more substituents independently selected from R40.
8. A pharmaceutical composition according to claim 7 wherein M is ArG1 or Het3, wherein the arylene or heteroarylene moieties are optionally substituted with one or more substituents independently selected from R40.
9. A pharmaceutical composition according to claim 8 wherein M is carbazolylene optionally substituted with one or more substituents independently selected from R40.
10. A pharmaceutical composition according to claim 9 wherein M is
Figure imgf000397_0001
11. A pharmaceutical composition according to any one of the claims 4 to4.10 wherein R40 is selected from -hydrogen, halogen, -CN, -CF3, -OCF3, -NO2, -OR41, -NR41R42, -SR41, -S(O)2R41,
-NR41C(O)R42, -OCi-C6-alkyl-C(O)NR41R42, -C2-C6-alkenyl-C(=O)OR41, -C(O)OR41, =0, -NH-C(=O)-O-CrC6-alkyl, or -NH-C(=O)-C(=O)-O-C1-C6-alkyl,
Ci-C6-alkyl or C2-C6- alkenyl which may each optionally be substituted with one or more substituents independently selected from R43,
• aryl, aryloxy, aryl-CrC6-alkoxy, aryl-Ci-C6-alkyl, aryl-C2-C6-alkenyl, heteroaryl, het- eroaryl-Ci-C6-alkyl, or heteroaryl-C2-C6-alkenyl, wherein the cyclic moieties optionally may be substituted with one or more substituents selected from R44.
12. A pharmaceutical composition according to any one of the claims 4 to 11 wherein R41 and R42 are independently selected from hydrogen, Ci-C6-alkyl, or aryl, wherein the aryl moieties may optionally be substituted with halogen or -COOH.
13. A pharmaceutical composition according to claim 4 to 12 wherein Q is a valence bond, -CH2-, -CH2-CH2-, -CH2-O-, -CH2-CH2-O-, -CH2-NH-, -CH2-CH2-NH-, -NH-CH2-, -NH-CH2- CH2-, -NH-C(=O)-, -C(=O)-NH-, -0-CH2-, -0-CH2-CH2-, or -C(=O)-.
14. A pharmaceutical composition according to any one of the claims 4 to 13 wherein R47 and R48 are independently selected from hydrogen, methyl and phenyl.
15. A pharmaceutical composition according to any one of the claims 4 to 14 wherein T is • hydrogen,
• C-i-Ce-alkyl optionally substituted with one or more substituents independently se¬ lected from R50,
• aryl, aryl-CrC6-alkyl, heteroaryl, wherein the alkyl, aryl and heteroaryl moieties are optionally substituted with one or more substituents independently selected from R50.
16. A pharmaceutical composition according to claim any one of the claims 4 to 15 wherein R50 is CrCe-alkyl, CrC6-alkoxy, aryl, aryloxy,
Figure imgf000398_0001
-C(=O)-NH- (CH2CH2O)mCi-C6-alkyl-COOH, aryl-CrC6-alkoxy , -OR51, -NO2, halogen, -COOH, -CF3, wherein any aryl moiety is optionally substituted with one or more R53.
17. A pharmaceutical composition according to any one of the claims 4 to 16 wherein m is 1 or 2.
18. A pharmaceutical composition according to any one of the claims 4 to 17 wherein R51 is methyl.
19. A pharmaceutical composition according to any one of the claims 4 to 18 wherein R53 is CrCe-alkyl, CrC6-alkoxy, -OR51, halogen.or -CF3.
20. A pharmaceutical composition according to any one of the claims 4 to 19 wherein R50A is -C(O)OCH3, -C(O)OCH2CH3 -COOH, -CH2C(O)OCH3, -CH2C(O)OCH2CH3, -CH2CH2C(O)- OCH3, -CH2CH2C(O)OCH2CH3, -CH2COOH, methyl, or ethyl.
21. A pharmaceutical composition according to any one of the claims 4 to 20 wherein R50B is -C(O)OCH3, -C(O)OCH2CH3 -COOH, -CH2C(O)OCH3, -CH2C(O)OCH2CH3, -CH2CH2C(O)- OCH3, -CH2CH2C(O)OCH2CH3, -CH2COOH, methyl, or ethyl.
22. A pharmaceutical preparation according to any one of the claims 1 to 21 wherein Frg1 consists of 0 to 5 neutral amino acids independently selected from the group consisting of GIy, Ala, Thr, and Ser.
23. A pharmaceutical preparation according to claim 22 wherein Frg1 consists of 0 to 5 GIy.
24. A pharmaceutical preparation according to any one of the claims 1 to 23 wherein GB is of the formula B1-B2-C(O)-, B1-B2-SO2- or B1-B2-CH2-, wherein B1 and B2 are as defined in claim 1.
25. A pharmaceutical preparation according to any one of the claims 1 to 23 wherein GB is of the formula B1-B2-C(O)-, B1-B2-SO2- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1.
26. A pharmaceutical preparation according to any one of the claims 1 to 23 wherein GB is of the formula B1-B2-C(O)-, B1-B2-CH2- or B1-B2-NH-, wherein B1 and B2 are as defined in claim 1.
27. A pharmaceutical preparation according to any one of the claims 1 to 23 wherein GB is of the formula B1-B2-CH2-, B1-B2-SO2- or B1-B2-NH-, wherein B1 and B2 are as defined in claim
1.
28. A pharmaceutical preparation according to any one of the claims 1 to 27 wherein B1 is a valence bond, -O-, or -S-.
29. A pharmaceutical preparation according to any one of the claims 1 to 27 wherein B1 is a valence bond, -O-, or -N(R68)-.
30. A pharmaceutical preparation according to any one of the claims 1 to 27 wherein B1 is a valence bond, -S-, or -N(R6B)-.
31. A pharmaceutical preparation according to any one of the claims 1 to 27 wherein B1 is .O-, -S- or -N(R6B)-.
32. A pharmaceutical preparation according to any one of the claims 1 to 31 wherein B2 is a valence bond, d-Cis-alkylene, C2-Ci8-alkenylene, C2-Ci8-alkynylene, arylene, heteroarylene, -CrCiβ-alkyl-aryl-^CC^-CrC^-alkyl-CC^-^CC^-Ci-Giβ-alkyl-O-d-dβ-alkyl-CC^)-, and the alkylene and arylene moieties are optionally substituted as defined in claim 1.
33. A pharmaceutical preparation according to any one of the claims 1 to 32 wherein the in- sulin is selected from the group consisting of human insulin, an analogue thereof, a deriva¬ tive thereof, and combinations of any of these.
34. A pharmaceutical preparation according to claim 33 wherein the insulin is human insulin.
35. A pharmaceutical preparation according to claim 33 wherein the insulin is an analogue of human insulin wherein position B28 is Asp, GIu, Lys, Leu, VaI or Ala.
36. A pharmaceutical preparation according to any one of the claims 33 to 35 wherein the insulin is an analogue of human insulin wherein position B29 is Pro, Asp or GIu.
37. A pharmaceutical preparation according to any one of the claims 33 to 36 wherein the insulin is an analogue of human insulin wherein position B9 is Asp or GIu.
38. A pharmaceutical preparation according to any one of the claims 33 to 37 wherein the insulin is an analogue of human insulin wherein position B10 is Asp or GIu.
39. A pharmaceutical preparation according to any one of the claims 33 to 38 wherein the insulin is an analogue of human insulin wherein position B1 is GIy.
40. A pharmaceutical preparation according to any one of the claims 33 to 39 wherein the insulin is an analogue of human insulin wherein position B3 is Lys, Thr, Ser, Ala or GIn.
41. A pharmaceutical preparation according to any one of the claims 33 to 40 wherein the insulin is an analogue of human insulin wherein position B25 is deleted.
42. A pharmaceutical preparation according to any one of the claims 33 to 41 wherein the insulin is an analogue of human insulin wherein position B27 is deleted.
43. A pharmaceutical preparation according to any one of the claims 33 to 42 wherein the insulin is an analogue of human insulin wherein position B30 is deleted.
44. A pharmaceutical preparation according to any one of the claims 33 to 43 wherein the insulin is an analogue of human insulin wherein position A18 is GIn.
45. A pharmaceutical preparation according to any one of the claims 33 to 44 wherein insulin is an analogue of human insulin wherein position A21 is Ala, GIn, GIu, GIy, His, lie, Leu, Met,
Phe, Ser, Thr, Trp, Tyr, VaI or hSer.
46. A pharmaceutical preparation according to any one of the claims 33 to 45 wherein the insulin is a derivative of human insulin or an analogue thereof having one or more lipophilic substituents.
47. A pharmaceutical preparation according to claim 46 wherein the Nε-amino group in posi¬ tion B29l_ys is modified by covalent acylation with a hydrophobic moiety such as an fatty acid derivative or an litocholic acid derivative.
48. A pharmaceutical preparation according to any one of the claims 1 to 47 wherein the ratio of the protamine-extended ligand of general formula (I) to zinc ion is 1 :20 to 20:1.
49. A pharmaceutical preparation according to any one of the claims 1 to 48 wherein the amount of zinc ions is 2-6 moles per mole of putative insulin hexamer.
50. A pharmaceutical preparation according to any one of the claims 1 to 49 wherein zinc ions are present in an amount corresponding to 10 to 40 μg Zn/100 U insulin.
51. A pharmaceutical preparation according to any one of the claims 1 to 50 wherein the ratio between insulin and the protamine-extended ligand according to any one of the claims 1 to 32 is in the range from 99:1 to 1 :99.
52. A pharmaceutical preparation according to any one of the claims 1 to 51 wherein the con- centration of insulin is 60 to 3000 nmol/ml.
53. A method of preparing a protamine-extended ligand according to claim 1 comprising the steps of
• Identifying starter compounds that binds to the R-state HisB10-Zn2+ site • optionally attaching a fragment consisting of 0 to 5 neutral α- or β-amino acids
• attaching protamine.
54. Method of prolonging the action of an insulin preparation which comprises adding a pro- tamine-extended ligand according to any one of the claims 1 to 32 to the insulin preparation.
55. A method of treating type 1 or type 2 diabetes comprising administering to a patient in need thereof a theraputically effective amount of a pharmaceutical preparation according to any one of the claims 1 to 52.
56. Use of a preparation according to any one of the claims 1 to 52 for the preparation of a medicament for treatment of type 1 or type 2 diabetes.
PCT/EP2005/053070 2004-07-09 2005-06-29 Phamaceutical preparations comprising insulin Ceased WO2006005683A1 (en)

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ITMI20110208A1 (en) * 2011-02-11 2012-08-12 Giuseppe Lembo HETEROCYCLES WITH ANTI-HYPERTENSIVE ACTIVITY
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US10865213B2 (en) 2016-02-16 2020-12-15 Massachusetts Institute Of Technology Max binders as MYC modulators and uses thereof
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WO2004056347A2 (en) * 2002-12-20 2004-07-08 Novo Nordisk A/S Pharmaceutical compositions comprising insulin and legends of insulin hexamer

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WO2000064940A1 (en) * 1999-04-27 2000-11-02 Eli Lilly And Company Insulin crystals for pulmonary administration
WO2003027081A2 (en) * 2001-09-14 2003-04-03 Novo Nordisk A/S NOVEL LIGANDS FOR THE HisB10 Zn2+ SITES OF THE R-STATE INSULIN HEXAMER
WO2004056347A2 (en) * 2002-12-20 2004-07-08 Novo Nordisk A/S Pharmaceutical compositions comprising insulin and legends of insulin hexamer

Cited By (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007119600A1 (en) * 2006-03-23 2007-10-25 Otsuka Pharmaceutical Co., Ltd. Carbazole compound
JP2007291079A (en) * 2006-03-27 2007-11-08 Otsuka Pharmaceut Co Ltd Pharmaceutical composed of carbostyril compound
WO2007135118A1 (en) * 2006-05-24 2007-11-29 Novo Nordisk A/S Soluble, stable insulin-containing formulations
US10259856B2 (en) 2008-03-18 2019-04-16 Novo Nordisk A/S Protease stabilized acylated insulin analogues
WO2012019106A3 (en) * 2010-08-06 2012-06-07 Board Of Regents Of The University Of Nebraska Compounds for use in modulating the nmda receptor
ITMI20110208A1 (en) * 2011-02-11 2012-08-12 Giuseppe Lembo HETEROCYCLES WITH ANTI-HYPERTENSIVE ACTIVITY
US10626095B2 (en) 2013-07-17 2020-04-21 Otsuka Pharmaceutical Co., Ltd. Cyanotriazole compounds
EP3230279A1 (en) * 2014-12-10 2017-10-18 Massachusetts Institute of Technology Fused 1,3-azole derivatives useful for the treatment of proliferative diseases
US10865213B2 (en) 2016-02-16 2020-12-15 Massachusetts Institute Of Technology Max binders as MYC modulators and uses thereof
US10265385B2 (en) 2016-12-16 2019-04-23 Novo Nordisk A/S Insulin containing pharmaceutical compositions
US10596231B2 (en) 2016-12-16 2020-03-24 Novo Nordisk A/S Insulin containing pharmaceutical compositions
CN111320681A (en) * 2020-01-23 2020-06-23 有机合成药品工业株式会社 High purity protamine sulfate
CN113929588A (en) * 2020-06-29 2022-01-14 沈阳化工研究院有限公司 Method for synthesizing 4-aminomethylbenzoic acid

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