HK1158619B - Diazepine and diazocane compounds as mc4 agonists - Google Patents

Description

Diazepanes and diazacyclooctanes as MC4 agonists

Technical Field

The present invention relates to diazepan and diazacyclooctane compounds, pharmaceutical compositions containing those compounds, and their use in therapy. The aforementioned compounds are useful as agonists at the melanocortin 4(MC4 or MCR4) receptor.

Background

Melanocortins are peptides derived from Proopiomelanocortin (POMC) that bind to and activate G-protein coupled receptors (GPCR's) of the melanocortin receptor family. Melanocortins regulate a variety of physiological processes including sexual function and behavior, food intake, and metabolism. Five melanocortin receptors, namely MCR1, MCR2, MCR3, MCR4, MCR5, have been cloned and expressed in a variety of tissues. MCR1 is specifically expressed in melanocytes and melanoma cells, MCR2 is the ACTH receptor and is expressed in adrenal tissue, MCR3 is most largely expressed in the brain and limbic system of the brain, MCR4 is widely expressed in the brain and spinal cord, and MCR5 is expressed in the brain and many peripheral tissues including skin, adipose tissue, skeletal muscle, and lymphoid tissues. MCR3 may be involved in the control of sexual function, food intake, and thermogenesis.

MCR4 is a G-protein-coupled seven transmembrane receptor that is expressed primarily in the hypothalamus, hippocampus, and thalamus (Gantz et al, 1993J Biol Chem 268: 15174-15179). This receptor is involved in central regulation of body weight: MCR4 is activated by alpha-Melanocyte Stimulating Hormone (MSH) derived from premanine and inactivated by acarus gene-associated protein (AGRP). In spiny mice, α -MSH causes weight loss, while abnormal expression of the spiny proteins leads to obesity (Fan et al, 1993 Nature 385: 165-. Additional evidence for the role of MCR4 in weight regulation comes from the mouse knockout model (Huszar et al, 1997 Cell 88: 131-141) and the human haplotype deficiency mutations (Valsse et al, 1998 Nat Genet 20: 113-114; Yeo et al, 1998 Nat Genet 20: 111-112; Hinney et al, 1999J Clin Endocrinol Metab 84: 1483-1486). In MCR4 knockout mice, increased body weight was recognized at 5 weeks of age. At 15 weeks of age, homozygous mutant females were on average twice as heavy as their wild-type littermates, while homozygous mutant males were-50% heavier than the wild-type controls. The MCR4 knockout mouse heterozygotes showed weight gains between those observed in wild-type and homozygous mutant littermates, confirming the gene dose effect of MCR4 ablation on weight regulation. The food intake of homozygous mutants was increased by-50% compared to wild-type inbreds (Huszar et al, 1997 Cell 88: 131-. [ from am.j.hum.genet., 65: 1501-1507, 1999]. MCR4 activation has been shown to cause penile erection in rodents, and MCR4 inactivation has been shown to cause obesity (reviewed in Hadley, 1999, Ann N Y Acad Sci., 885: 1-21, Wikberg et al, 2000, Pharmacol Res., 42(5), 393-420).

Chaki and Nakazato in Drugs Of The Future, 2004, 29 (10): 1065-1074 relates to the possible therapeutic use of ligands acting at the MC4 receptor. Diazepan derivatives are reported in WO95/00497, WO 97/17973, WO 98/07692, WO 98/20001, WO2006/040192 and EP 1867639. Inhibitors of FXa are reported in WO 98/54164. Compounds useful for the treatment of bone defect disorders are reported in WO 99/42107. Antagonists of the H3 receptor are reported in WO 02/072570. Modulators of PPAR are reported in US 2005/0234046.

The compounds of the invention are useful for treating diseases, disorders, or conditions responsive to activation of the MC4 receptor, including:

male and female sexual dysfunction, including hypoactive sexual desire disorder, orgasmic disorder and/or female pain disorder during sexual intercourse, male erectile dysfunction;

obesity (through appetite decline, increased metabolic rate, decreased fat intake or decreased craving); and

diabetes (by increasing glucose tolerance and/or decreasing insulin resistance).

The compounds of the invention are useful in the treatment of other diseases, disorders, or conditions, including, but not limited to, hypertension, hyperlipidemia, osteoarthritis, cancer, gallbladder disease, sleep apnea (sleep apnea), depression, anxiety, obsessive-compulsive disorders, neurasthenia, insomnia/sleep disorders, substance abuse (substention abuse), pain, fever, inflammation, immunomodulation, rheumatoid arthritis, tanning (skinning), acne and other skin conditions, neuroprotection and cognitive and memory enhancement (including treatment of alzheimer's disease), treatment of lower urinary tract dysfunction (including urinary incontinence-overactive bladder, urinary frequency, nocturia, urgency, urinary incontinence (any condition in which there is involuntary leakage of urine, including stress, urge and mixed urinary incontinence), Urinary incontinence associated overactive bladder, nocturia, primary nocturnal enuresis, continuous urinary incontinence, episodic urinary incontinence such as incontinence during sexual intercourse and Lower Urinary Tract Syndrome (LUTS) associated with Benign Prostatic Hypertrophy (BPH), as well as any of the indications of the books in the previously cited patent applications.

The compounds of the present invention are particularly useful in the treatment of female sexual dysfunction, male erectile dysfunction, obesity, diabetes and lower urinary tract dysfunction conditions.

The term "treatment" as used herein is intended to encompass both prevention and management, i.e., prophylactic and palliative treatment of the indicated condition.

Desirable characteristics of MCR4 agonist compounds of the invention include: desirable MCR4 agonist potency as detailed below; selectivity for MCR4 agonism relative to MCR1 and/or MCR5 and/or MCR3, as detailed below; both desirable MC4R agonist potency and selectivity for MCR4 over MCR1 and/or MCR5 and/or MCR 3; good biopharmaceutical properties such as victory stability, solubility, oral bioavailability, proper metabolic stability.

Disclosure of Invention

According to one embodiment, the present invention relates to a compound of formula (I):

wherein n and R¹、R²、R³L and R⁴As defined hereinafter in the detailed description.

Another embodiment of the present invention is directed to a pharmaceutical composition comprising a compound of formula (I). In one aspect, the composition comprises a therapeutically effective amount of a compound of formula (I). In another aspect, the composition may further comprise one or more additional pharmaceutical agents (e.g., those described below).

Yet another embodiment of the present invention is directed to a method of treating a condition (including a disease and/or disorder) benefiting from the agonism of MC4, comprising administering to a subject in need of such treatment a therapeutically effective amount of a compound of formula (I) (or a pharmaceutical composition thereof). In one aspect, the disorder is Female Sexual Dysfunction (FSD), male erectile dysfunction, or obesity.

Detailed Description

The invention relates to compounds of formula (I)

Or pharmaceutically acceptable salts, solvates (including hydrates) and prodrugs thereof, wherein

n is 0 or 1;

R¹is- (C)₁-C₄) Alkyl or Het¹；

R²Is phenyl or pyridyl, wherein said phenyl or pyridyl is optionally substituted with 1 to 3 substituents independently selected from halogen, CN, - (C)₁-C₄) Alkyl and- (C)₁-C₄) Substituent of alkoxy, wherein- (C)₁-C₄) Alkyl and- (C)₁-C₄) Alkoxy groups are optionally substituted with 1 to 3 fluorine atoms;

R³is phenyl or pyridyl, wherein said phenyl or pyridyl is optionally substituted with 1 to 3 substituents independently selected from halogen, CN, - (C)₁-C₄) Alkyl and- (C)₁-C₄) Substituent of alkoxy, wherein- (C)₁-C₄) Alkyl and- (C)₁-C₄) Alkoxy groups are optionally substituted with 1 to 3 fluorine atoms;

l is-CO-and R⁴Is- (C)₁-C₄) Alkyl, - (C)₁-C₄) Alkoxy, - (C)₃-C₆) Cycloalkyl, - (C)₁-C₂) Alkyl radical (C)₃-C₆) Cycloalkyl, - (C)₁-C₂) Alkyl radical (C)₁-C₄) Alkoxy, -NH₂、-NH(C₁-C₄) Alkyl, -N [ (C)₁-C₄) Alkyl radical]₂Or Het²Wherein said- (C)₁-C₄) Alkyl groups optionally substituted with 1 to 3 fluorine atoms and wherein- (C)₃-C₆) Cycloalkyl radicals optionally substituted by 1 to 3 fluorine atoms or- (C)₁-C₄) Alkyl groups;

or L is-SO₂-and R⁴Is- (C)₁-C₄) Alkyl, - (C)₃-C₆) Cycloalkyl, - (C)₁-C₂) Alkyl radical (C)₃-C₆) Cycloalkyl, - (C)₁-C₂) Alkyl radical (C)₁-C₄) Alkoxy, -NH₂、-NH(C₁-C₄) Alkyl, -N [ (C)₁-C₄) Alkyl radical]₂Or Het²；

Het¹Is that

(i) A 6-membered ring containing 1 or 2N atoms, wherein the ring is aromatic or the ring contains 2 intra-ring bisA bond and an ═ O substituent, said ring being optionally substituted with 1 to 3 substituents independently selected from halogen, CN and — (C)₁-C₄) Alkyl is substituted by a substituent;

(ii) a 6-membered aromatic ring comprising 1 or 2N atoms, the 6-membered aromatic ring being fused at the 3, 4 position relative to the bond to the pyrrolidine ring to a 5-membered aromatic ring comprising 1 to 3 additional N atoms; or

(iii) A tetrahydropyranyl group;

Het²is that

(i) A 5-membered aromatic ring containing 1 or 2N atoms and optionally also O, S or N atoms,

(ii) a 4-to 6-membered saturated ring comprising 1N atom; or

(iii) A 6-membered saturated ring containing one O atom and optionally also an N atom.

The term "alkyl" refers to a straight or branched chain saturated aliphatic hydrocarbon group containing a specified number of carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl.

The term "alkoxy" refers to the group OR, wherein R is alkyl as defined above.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "cycloalkyl" refers to a monocyclic aliphatic alkyl group containing the specified number of carbon atoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

Examples of "a 6-membered ring containing 1 or 2N atoms, wherein the ring is aromatic or the ring contains 2 intra-ring double bonds and an ═ O substituent" include pyrazole, pyridine, pyrazine, pyrimidine, pyridazine and pyridazinone.

Examples of "a 6-membered aromatic ring containing 1 or 2N atoms, said 6-membered aromatic ring being fused at the 3, 4-position relative to the binding to the pyrrolidine ring with a 5-membered aromatic ring containing 1 to 3 additional N atoms" include imidazo [1, 2-b ] pyridazine and [1,2,4] triazolo [4,3-b ] pyridazine.

Examples of "a 5-membered aromatic ring containing 1 or 2N atoms and optionally also O, S or N atoms" include pyrrole, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, triazole, oxadiazole and thiadiazole.

Examples of "a 4 to 6 membered saturated ring containing 1N atom" include azetidine, pyrrolidine, piperidine and piperazine.

Examples of "a 6-membered saturated ring containing one O atom and optionally also an N atom" include tetrahydropyran and morpholine.

In one embodiment, n is 1.

In one embodiment, R¹Is- (C)₁-C₄) An alkyl group. In another embodiment, R¹Is a tert-butyl group.

In one embodiment, R¹Is Het¹Wherein Het is¹Is (i) a 6-membered ring containing 1 or 2N atoms, wherein the ring is aromatic or the ring contains 2 intra-ring double bonds and ═ O substituents, the ring being optionally substituted by a substituent selected from the group consisting of halogen, CN and ═ C₁-C₄) Alkyl is substituted by a substituent; or (ii) a 6-membered aromatic ring comprising 1 or 2N atoms, the 6-membered aromatic ring being fused to a 5-membered aromatic ring comprising 1 or 2 additional N atoms at the 3, 4 position relative to the bond to the pyrrolidine ring.

In another embodiment, R¹Is Het¹Wherein Het is¹Is pyridin-2-yl, pyridin-3-yl, pyridazin-3-yl, 6-oxo-1, 6-dihydropyridazin-3-yl, 6-oxo-1, 6-dihydropyridin-3-yl, 2-oxo-1, 2-dihydropyrimidin-4-yl, 6-oxo-1, 6-dihydropyrimidin-4-yl, 2-oxo-1, 2-dihydropyridin-4-yl, [1,2,4] dihydropyridin-4-yl]Triazolo [4,3-b]Pyridazin-6-yl or 6-oxo-1, 6-dihydropyridin-2-yl, optionally substituted by one or two groups independently selected from- (C)₁-C₄) Substituents for alkyl, halogen and CNAnd (4) substituting.

In yet another embodiment, R¹Is Het¹Wherein Het is¹Is 6-oxo-1, 6-dihydropyridazin-3-yl, 1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl or [1,2,4]]Triazolo [4,3-b]Pyridazin-6-yl.

In one embodiment, R²Is phenyl or pyridyl, wherein said phenyl or pyridyl is optionally substituted by 1 or 2 substituents independently selected from halogen, CN, - (C)₁-C₄) Alkyl and- (C)₁-C₄) Substituted by a substituent of alkoxy. In another embodiment, R²Is 2, 4-difluorophenyl, 2-fluoro-4-methoxy, 4-cyanophenyl or 5-chloropyridin-2-yl.

In one embodiment, R³Is optionally substituted by 1 or 2 substituents independently selected from halogen and (C)₁-C₄) Phenyl substituted with a substituent of alkoxy. In another embodiment, R³Is 4-chlorophenyl.

In one embodiment, L is-CO-and R⁴Is optionally substituted by 1 to 3 fluorine atoms — (C)₁-C₄) Alkyl, - (C)₁-C₄) Alkoxy, optionally substituted by 1 or 2 fluorine atoms or- (C)₁-C₄) Alkyl group substituted- (C)₃-C₆) Cycloalkyl, - (C)₁-C₂) Alkyl radical (C)₃-C₆) Cycloalkyl, - (C)₁-C₂) Alkyl radical (C)₁-C₄) Alkoxy, -NH (C)₁-C₄) Alkyl, -N [ (C)₁-C₄) Alkyl radical]₂Or Het²Wherein Het is²Is a 5-membered aromatic ring containing 2N atoms or a 6-membered saturated ring containing one O atom and another optional N atom.

In another embodiment, L is-CO-and R⁴Is- (C)₁-C₄) Alkyl or- (C)₁-C₄) Alkoxy, wherein said- (C)₁-C₄) The alkyl group is optionally substituted with 1 to 3 fluorine atoms.

It is to be understood that the present invention encompasses all combinations of the particular embodiments described above that conform to the definition of the compounds of formula (I).

Representative compounds of the invention include:

6- [ (3S,4R) -3- { [5S- (4-chlorophenyl) -4- (3,3, 3-trifluoropropionyl) -1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-1-yl ] -2-methylpyridazin-3 (2H) -one;

6- [ (3S,4R) -3- { [5S- (4-chlorophenyl) -4-isobutyryl-1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (2, 4-difluorophenyl) pyrrolidin-1-yl ] -2-methylpyridazin-3 (2H) -one;

6- [ (3S,4S) -3- { [5S- (4-chlorophenyl) -4-isobutyryl-1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (5-chloropyridin-2-yl) pyrrolidin-1-yl ] -2-methylpyridazin-3 (2H) -one;

8- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

8S- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

8R- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

8R- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

6- [ (3S,4R) -3- { [ 4-acetyl-5S- (4-chlorophenyl) -1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-1-yl ] pyridazin-3 (2H) -one;

8S- (4-chlorophenyl) -4- { [ (3S,4S) -4- (5-chloropyridin-2-yl) -1- (6-cyanopyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

1- { [ (3S,4S) -1-tert-butyl-4- (5-chloropyridin-2-yl) pyrrolidin-3-yl ] carbonyl } -5S- (4-chlorophenyl) -4-isobutyryl-1, 4-diazacyclooctane;

6- [ (3S,4S) -3- { [ 4-acetyl-5S- (4-chlorophenyl) -1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (5-chloropyridin-2-yl) pyrrolidin-1-yl ] [1,2,4] triazolo [4,3-b ] pyridazine;

8S- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2, 4-difluorophenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

or pharmaceutically acceptable salts, solvates (including hydrates), and prodrugs thereof.

Pharmaceutically acceptable salts of the compounds of formula (I) include the acid addition and base salts thereof.

Suitable acid addition salts are formed from acids which form non-toxic salts. Examples include acetate, adipate, aspartate, benzoate, benzenesulfonate, bicarbonate/carbonate, bisulfate/sulfate, borate, camsylate, citrate, cyclamate, edisylate, ethanesulfonate, formate, fumarate, glucoheptonate, gluconate, glucuronic acid, hexafluorophosphate, oxybenzoyl benzoate, hydrochloride/chloride, hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malic acid, maleic acid, malonate, methanesulfonate, methylsulfate, naphthenate, 2-naphthalenesulfonate, nicotinate, nitrate, orotate, oxalate, palmitic acid, pamoate, sodium/hydrogen/dihydrogen phosphate, pyroglutamate, dihydrogenphosphate, dihydrogen, Sucrose salts, stearates, succinates, tannates, tartrates, tosylates, trifluoroacetates and oxynaphthalene carboxylates.

Suitable base salts are formed from bases which form non-toxic salts. Examples include aluminum, arginine, benzathine, calcium, choline, diethylamine, dialcohol, glycine, lysine, magnesium, meglumine, oleylamine, potassium, sodium, tromethamine and zinc salts.

Hemisalts of acids and bases, such as hemisulfate and hemicalcium salts, may also be formed.

For a review of suitable salts, see Stahl and WermuthHandbook of Pharmaceutical Salts：Properties，Selection，and Use(Wiley-VCH, 2002), which is incorporated herein by reference.

The compounds of the present invention may exist in solid state from completely amorphous to completely crystalline. The term "amorphous" refers to the following state: where the substance lacks a wide range of order at the molecular level and may exhibit physical properties of a solid or liquid depending on temperature. Typically, such materials do not provide a unique X-ray diffraction pattern, and while exhibiting the properties of a solid, they are described more as liquids. When heated, a change occurs from a solid to a liquid property, which is characterized by a change of state, usually two-stage ("glass transition"). The term "crystallization" refers to the following solid phase: wherein the substance has a regular ordered internal structure on a molecular level and provides an X-ray diffraction pattern having specific peaks. Such materials will also exhibit the properties of a liquid when heated sufficiently, but the change from solid to liquid is characterized by a phase change, typically first order ("melting point").

The compounds of the present invention may also exist in unsolvated and solvated forms. The term "solvate" is used herein to describe a molecular complex comprising a compound of the invention and one or more pharmaceutically acceptable solvent molecules, such as ethanol. When the solvent is water, the term "hydrate" is used.

One generally accepted classification system for organic hydrates is the following: which define isolated sites, channels or hydrates of coordination of metal ions-see k.rPolymorphism in Pharmaceutical Solids(H.G. Brittain eds., Marcel dekker, 1995), which is incorporated herein by reference. An isolated site hydrate is a hydrate of: wherein water molecules are isolated from direct contact with each other by intervening organic molecules. In channel hydrates, water molecules are present in lattice channels where they are next to other water molecules. In the metal ion coordinated hydrate, water molecules are combined with the metal ions.

When the solvent or water is tightly bound, the complex will have a well-defined stoichiometry independent of humidity. However, when the solvent or water is weakly bound, as in channel solvates and hygroscopic compounds, the water/solvent content will depend on the humidity and drying conditions. In this case, non-stoichiometry would be normal.

Also included within the scope of the invention are multicomponent complexes (other than salts and solvates) in which the drug and at least one other component are present in stoichiometric or non-stoichiometric amounts. Complexes of this type include clathrates (drug-matrix inclusion complexes) and co-crystals. The latter is generally defined as a crystalline complex of neutral molecular components held together by non-covalent interactions, but may also be a complex of a neutral molecule and a salt. Co-crystals can be prepared by melt crystallization, by recrystallization from a solvent, or by physical milling of the various components together-see o.almarsson and m.j.zawortko Chem commu,171889-18962004), which is incorporated herein by reference. For a general review of multicomponent complexes, see J Pharm Sci by Haleblian,64(8) 1269-1288 (8 months 1975), which is incorporated herein by reference.

The compounds of the invention may also be present in the mesomorphic state (mesomorphic or liquid crystal) when subjected to suitable conditions. A mesomorphic state is an intermediate state between a true crystalline state and a true liquid state (melt or solution). The mesogenic phenomenon due to a change in temperature is called "thermotropic liquid crystal", and the mesogenic phenomenon due to the addition of, for example, water or another solvent is called "lyotropic liquid crystal". Compounds with the potential to form lyotropic liquid crystals are called "amphiphilic" and consist of compounds with ionic polarityTerminal group (such as-COO)^-Na⁺、-COO^-K⁺or-SO₃ ^-Na⁺) Or non-ionic polar end groups (such as-N)^-N⁺(CH₃)₃) The molecular composition of (a). For more information see n.h.hartshorn and a.stuartCrystals and the Polarizing MicroscopeFourth edition (EdwardAlnold, 1970), which is incorporated herein by reference.

All references hereinafter to compounds of formula (I) include references to salts, solvates, multicomponent complexes and liquid crystals thereof, and to solvates, multicomponent complexes and liquid crystals thereof.

The compounds of the present invention include compounds of formula (I) as hereinbefore defined, including all polymorphs and crystal forms thereof, prodrugs and isomers thereof (including optical, geometric and tautomeric isomers), and isotopically-labelled compounds of formula (I) as hereinafter defined. As indicated, so-called "prodrugs" of the compounds of formula (I) are also within the scope of the present invention. Thus, certain derivatives of the compounds of formula (I) which have no or little pharmacological activity themselves can be converted, for example by hydrolytic cleavage, into compounds of formula (I) having the desired activity when administered into or onto an organism. Such derivatives are referred to as "prodrugs". Further information on prodrug use can be seenPro-drugs as Novel Delivery SystemsVol.14, ACSSymposium Series (T.Higuchi and W.Stella) andBioreversible Carriers in Drug Designpergamon Press, 1987(e.b. roche eds., american pharmaceutical Association), which is incorporated herein by reference.

Prodrugs according to the invention can be prepared, for example, by replacing suitable functional groups present in the compounds of formula (I) with certain molecular moieties known to those skilled in the art as "precursor-moieties", such as, for example, those of hDesign of Prodrugs(Elsevier, 1985), which is incorporated herein by reference.

In addition, certain compounds of formula (I) may themselves act as prodrugs of other compounds of formula (I).

The compounds of formula (I) may have asymmetric carbon atoms. The bond formed by an asymmetric carbon in the compound of the present invention may be represented by a solid (—), a wedge-shaped solid, or the likeOr dotted wedgeTo be depicted. The use of a solid line to depict the bond to an asymmetric carbon atom is intended to represent all possible stereoisomers contained at that carbon atom. The use of solid or dashed wedges to depict bonds to asymmetric carbon atoms is intended to mean that only the stereoisomers shown are intended to be included. It may be that the compound of formula (I) may contain more than one asymmetric carbon atom. In those compounds, the use of a solid line to depict bonds made by asymmetric carbon atoms is intended to indicate that all possible stereoisomers are intended to be encompassed. When the compounds of formula (I) contain an alkenyl or alkenylene group, geometric cis/trans (or Z/E) isomers are possible. Tautomerism ("tautomerism") can occur when structural isomers can interconvert through a low energy barrier. This phenomenon can occur in proton tautomerism in compounds of formula (I) containing, for example, imino, keto, or oxime groups, or in so-called valence tautomerism in compounds containing aromatic moieties. A single compound may then exhibit more than one isomerism. As an example to illustrate the tautomeric relationship, the following compounds (wherein, for example, the "Het" is mentioned¹The "groups shown below)," keto "and" enol "tautomers are all encompassed by" Het "of the compounds of the invention¹"in the range of):

all stereoisomers, geometric isomers and tautomeric forms of the compounds of formula (I) are included within the scope of the invention, including compounds exhibiting more than one isomerism, and mixtures of one or more thereof. Also included are acid addition salts or base salts (e.g., d-lactate or l-lysine salts) or racemates (e.g., dl-tartrate or dl-arginine salt) in which the counter ion is optically active.

The cis/trans isomers can be isolated by conventional methods well known to those skilled in the art, such as chromatography and fractional crystallization.

Conventional methods for the preparation/separation of individual enantiomers include chiral synthesis from suitable optically pure precursors or resolution of the racemate (or the racemate of a salt or derivative) using, for example, chiral High Pressure Liquid Chromatography (HPLC).

Alternatively, the racemate (or a racemic precursor) may be reacted with a suitable optically active compound, for example, an alcohol, or, in the case of compounds of formula (I) comprising an acid or base moiety, a base or acid (such as 1-phenylethylamine or tartaric acid). The resulting diastereomeric mixtures can be separated by chromatography and/or fractional crystallization and one or both of the diastereomers converted to the corresponding pure enantiomers by means well known to those skilled in the art.

Chiral compounds of the invention (and chiral salts thereof) can be obtained in enantiomerically enriched form using chromatography (typically HPLC) on an asymmetric resin in a mobile phase consisting of a hydrocarbon, typically heptane or hexane containing 0-50% by volume isopropanol (typically 2-20%) and may contain 0-5% by volume alkylamine. The eluate is concentrated to give an enriched mixture. The absolute composition of the mobile phase will depend on the chiral stationary phase (asymmetric resin) chosen.

Stereoisomeric aggregates can be separated by conventional techniques known to those skilled in the art-see, e.g., e.l.eliel and s.h.wilenStereochemistry of Organic Compounds(Wiley, New York, 1994), which is incorporated herein by reference.

The present invention includes all pharmaceutically acceptable isotopically-labeled compounds of formula (I) wherein one or more atoms are replaced by an atom having the same atomic number, but an atomic mass or mass number different from the atomic mass or mass number which predominates in nature.

Examples of isotopes suitable for inclusion in the compounds of the invention include isotopes of the following elements: hydrogen, such as²H and³h; carbon, such as¹¹C、¹³C and¹⁴c; chlorine, e.g. of³⁶Cl; fluorine, such as¹⁸F; iodine, such as¹²³I and¹²⁵i; nitrogen, e.g.¹³N and¹⁵n; oxygen, such as¹⁵O、¹⁷O and¹⁸o; phosphorus, e.g.³²P; and sulfur, such as³⁵S。

The following schemes, including those described in the examples and preparations, illustrate the synthesis of compounds of formula (I). Those skilled in the art will appreciate that the compounds of the present invention and intermediates thereof can be prepared by methods other than those specifically described herein, e.g., by variations of the methods described herein, e.g., by methods known in the art. Examples of suitable guidelines for synthesis, interconversion of functional groups, use of protecting groups, etc., are: RC Larock, "Comprehensive Organic Transformations", VCH Publishers Inc. (1989); advanced Organic Chemistry "by March, Wiley Interscience (1985); s Warren, "design Organic Synthesis", Wiley Interscience (1978); s Warren, "Organic Synthesis-The DisconnectionAproach", Wiley Interscience (1982); RK Mackie and DM Smith, "Guideboost to Organic Synthesis", Longman (1982); TW Greene and PGM Wuts, "Protective Groups in Organic Synthesis", John Wiley and sons, Inc. (1999); and PJ, "Protecting Groups" by Kocienski, Georg Thieme Verlag (1994).

In the following general synthetic methods, the substituents R, unless otherwise indicated, are¹、R²、R³、R⁴And L is as hereinbefore defined for the compound of formula (I) above.

The scheme illustrates the preparation of compounds of formula (I) by acylation of intermediate (II) with an acylating agent (III).

Scheme 1

Typical conditions include stirring a solution of diazepan or diazacyclooctane of the formula (II) and an acylating agent of the formula (III) with a base in a suitable solvent at room temperature. Suitable acylating agents (III) include carboxylic acid chlorides, sulfonyl chlorides, carbamoyl chlorides and chloroformates, and are either commercially available or known to those skilled in the art by reference to the prior art.

Scheme 2 illustrates an alternative preparation method for the preparation of certain compounds of general formula (I) wherein L is a carbonyl group from diazepan and diazacyclooctane intermediates (II) using a peptide coupling reagent (IV).

Scheme 2

Typical conditions include stirring a solution of diazepan or diazacyclooctane of formula (II) and a carboxylic acid of formula (IV), and 1- (3-dimethylaminopropyl) -3-ethyl-carbodiimide hydrochloride (EDCI) or its methiodide salt plus triethylamine and 1-hydroxybenzotriazole hydrate (HOBt) in Dichloromethane (DCM). The carboxylic acids of formula (IV) are commercially available or known to those skilled in the art by reference to the prior art. Another alternative suitable method is to stir a solution of the intermediate compound of formula (II) and the acid of formula (IV) in an inert solvent with a suitable peptide coupling reagent and, if necessary, add a suitable base and/or additive. Suitable peptide coupling reagents include O-benzotriazol-1-yl-N, N '-tetramethyluronium Hexafluorophosphate (HBTU), 2- { 1H-benzotriazol-1-yl } -1, 1,1, 3-tetramethyluronium tetrafluoroborate (TBTU), O- (7-azabenzotriazol-1-yl) -N, N' -tetramethyluronium Hexafluorophosphate (HATU), 2-chloro-1, 3-dimethylimidazoline hydrochloride (DIC), 1-propylphosphoric acid cyclic anhydride (T3P), or polymer-supported Mukaiyama reagent; and suitable bases include pyridine, triethylamine, diisopropylethylamine, N-methylmorpholine or dimethylaminopyridine. Any suitable inert solvent may be used in place of the above, wherein inert solvent means a solvent that does not contain a carboxylic acid or a primary or secondary amine. At least one equivalent of each coupling reagent should be used, and one or both may be used in excess if desired.

Scheme 3 illustrates an alternative route to prepare compounds of general formula (I) from functionalized diazepanes and diazacyclooctanes of general formula (V) and pyrrolidines of general formula (VI) using peptide coupling reagents as described in scheme 2.

Scheme 3

Scheme 4 further illustrates the preparation of R wherein R is prepared by a protecting group strategy¹Is Het¹Alternative routes to the compounds of the general formula (I)

Scheme 4

PG¹Are suitable nitrogen protecting groups.

PG²Is LR⁴Or with PG¹Orthogonal to the other nitrogen protecting group.

In scheme 4, diazepan and diazacyclooctane intermediates of general formula (VII) are coupled with protected pyrrolidinic acid intermediates of general formula (VIII) using standard peptide coupling procedures as previously described in schemes 2 and 3 to provide coupled intermediates containing orthogonal protection of general formula (IX). The nitrogen-protecting group PG may be protected using standard deprotection strategies¹And PG²Separately removed to provide intermediates of formula (X) (via PG)¹By deprotection) or intermediates of the general formula (XII) (by PG)²Deprotection of (ii). Any suitable nitrogen Protecting group may be used (as described in t.w.greene and p.g.wuts, third edition, "Protecting Groups in organic synthesis", Wiley-Interscience, 1999). Typical nitrogen Protecting Groups (PG) suitable for use herein include t-butoxycarbonyl (t-Boc), which can be readily removed by treatment with an acid such as trifluoroacetic acid or hydrochloric acid in an organic solvent such as dichloromethane or 1, 4-dioxane; benzyl groups, which can be easily removed by hydrogenation in the presence of a suitable catalyst or by treatment with 1-chloroethyl chloroformate (ACE-Cl) followed by methanolysis; or tert-butyl, which can also be easily removed by ACE-Cl and methane decomposition.

R can be introduced by displacement of a suitable leaving group (leaving group displacement ")¹Group (herein R)¹Het as described above¹) For example, of the formula "Het¹Suitable heteroaromatic precursors "Het" of-Z¹-Z ", wherein Z is a suitable leaving group. Suitable leaving groups include halogen. In some cases, transition metal catalysis (e.g., palladium, copper), optionally in combination with a phosphine ligand (such as binaphthyl-2, 2' -diyl bis-phenylphosphine), may be required to obtain the desired coupled product.

According to scheme 4, intermediates of formula (X) can be subjected to leaving group displacement ("metatated") to give intermediates of formula (XI), which can be prepared by reacting PG with a leaving group²Deprotection followed by R following the procedures described in schemes 1 and 2⁴L terminates the exposed NH functionality to produce a compound of formula (I). Alternatively, intermediates of general formula (XII) can be prepared with R following the procedures described in schemes 1 and 2⁴L is terminated and then PG is passed¹And subsequently subjected to leaving group displacement ("metathesis") as described above to give compounds of the general formula (I).

Or alternatively, wherein R¹Is a designated Het¹Compounds of the general formula (I) of the radicals can be converted into compounds in which R¹Is a different Het¹Other compounds of the general formula (I). For example:

i) wherein Het can be hydrolyzed under acidic or basic conditions¹Compounds of formula (Ia) containing a suitable leaving group Z, such as chloro or methoxy, are converted to compounds of formula (Ib) as shown in scheme 5. Acidic conditions are preferred, and the treatment of the compound of formula (Ia) with acetic acid at reflux temperature is particularly preferred. Or alternatively, a compound of formula (Ia) wherein Z is chloro may be reacted with a compound of formula Y-O^-To obtain an intermediate of formula (Ia) wherein Z is OY. Followed by hydrolysis to give the compound of formula (Ib). Suitable Y groups may include methyl or benzyl.

Scheme 5

ii) wherein Het is obtainable by treatment with a base and an alkylating agent in a suitable solvent¹As shown in scheme 6 and R⁵Conversion of a compound of formula (Ic) which is H to wherein R⁵A compound of formula (Id) which is an alkyl group. Suitable bases include sodium hydride, lithium diisopropylamide, and sodium bis (trimethylsilyl) amide; suitable alkylating agents include methyl iodide, tosylate, dimethyl sulfate, and ethyl iodide; and suitable solvents include tetrahydrofuran, dimethylformamide and N-methyl-2-pyrrolidone. In the reactionOptional additives such as lithium salts (e.g., lithium bromide) may also be present in the mixture.

Scheme 6

The pyrrolidines of the general formulae (VI) and (VIII) can be prepared by de-esterifying the precursor esters of the general formulae (XIV) and (XV), respectively, using a variety of known literature methods as shown in scheme 7.

Scheme 7

Preferred methods include alkaline hydrolysis of esters of formula (XIV) and (XV) using aqueous solutions of suitable metal hydroxides in suitable co-solvents. Suitable metal hydroxides include those derived from alkali metals (e.g., Li, Na or K) or alkaline earth metals (e.g., Ca or Ba), and suitable cosolvents include water-miscible organic solvents such as THF, dioxane, and hydroxyl solvents (e.g., methanol and ethanol). Another preferred method of deesterifying the esters of the general formulae (XIV) and (XV) is by treatment with potassium trimethylsilanolate in a suitable solvent such as acetonitrile or toluene.

Scheme 8 illustrates a route to prepare novel pyrrolidine ester intermediates from trans cinnamate derivatives (XVI).

Scheme 8

PG¹Are suitable protecting groups for the purpose of protecting,such as tert-butyl or benzyl.

Xc is a chiral auxiliary moiety.

Cinnamic acid (XVI) is commercially available or known to those skilled in the art by reference to the prior art. Cinnamic acid (XVI) can be coupled to a variety of chiral auxiliary moieties (Xc) known in the literature using standard peptide coupling reagents as described in scheme 2 to give homochiral cinnamate derivatives of general formula (XVII). In this regard, commercially available oxazolidinone chiral auxiliary moieties are preferred. Intermediate (XVII) undergoes a [3+2] cycloaddition with an azomethine ylid precursor of formula (XX) to provide a racemic pyrrolidine of formula (XVIII) in which the trans stereochemistry predominates or is entirely trans stereochemistry. The reaction requires an inert solvent such as dichloromethane or toluene or tetrahydrofuran and requires activation of one or more of the following: (1) acid catalysts, such as TFA; (2) desilication agents such as silver fluoride; (3) and (4) heating. The racemic compound of formula (XVIII) can be resolved by standard methods such as chromatography or fractional crystallization to give the homochiral intermediate of formula (XIX). The chiral auxiliary moiety Xc comprised in the compound of formula (XIX) can be separated using methods in the prior art to obtain a pyrrolidine ester of formula (XV). In particular, oxazolidinone chiral auxiliary moieties (chiralities) can be deprotected with a lewis acid such as samarium triflate in methanol.

The pyrrolidine ester of formula (XIV) can be prepared from a pyrrolidine ester of formula (XV) by a deprotection and leaving group displacement ("hethylation") strategy as described in scheme 4.

Scheme 9 illustrates the preparation of intermediates of formula (II) from diazepane and diazacyclooctane intermediates of formula (XXI) by coupling with a pyrrolidinic acid of formula (VI).

Scheme 9

The diazepan and diazacyclooctane intermediates of formula (XXI) can be regioselectively coupled with the aforementioned pyrrolidinic acid of formula (VI) under peptide coupling conditions as described in scheme 2 to give intermediates of formula (II).

Scheme 10 illustrates intermediates of formula (XXII) (wherein PG³Is a nitrogen protecting group such as benzyl or tert-butoxycarbonyl) to prepare intermediates of general formula (XXIII).

Scheme 10

Compounds of general formula (XXIII) can be prepared by acylation (as described in scheme 1) or by using peptide coupling reagents (as described in scheme 2). There are several methods that can be used to prepare the precursors of formula (XXII) including, but not limited to, regioselective mono-protection of compounds of formula (XXI) as exemplified in preparation 13 or more direct assembly as exemplified in preparation 2.

The intermediate compounds of the formulae (II), (V), (VI), (VII), (VIII), (IX), (XI), (XII), (XIII), (XIV), (XV), (XXI), (XXII) and (XXIII) described above represent further embodiments of the present invention.

Chiral compounds of the invention (and chiral precursors thereof) can be obtained in enantiomerically enriched form using chromatography (typically HPLC) on an asymmetric resin with a mobile phase consisting of a hydrocarbon, typically heptane or hexane with 0-50% by volume isopropanol (typically 2-20%) and possibly 0-5% by volume alkylamine. The eluate is concentrated to give an enriched mixture. The absolute composition of the mobile phase will depend on the chiral stationary phase (asymmetric resin) chosen.

The skilled person will appreciate that in addition to protecting nitrogen or acid groups as described hereinbefore, at various points in the synthesis of compounds of formula (I), it may be necessary to protect other groups, such as, for example, hydroxyl groups, with a suitable protecting group, followed by removal of the protecting group. The method of deprotection of any particular group will depend on the protecting group. For example, see TW Greene and PGM Wutz for "Protective groups in organic synthesis", which is incorporated herein by reference. For example, when the hydroxyl group is protected as a methyl ester, the deprotection conditions may, for example, comprise refluxing in 48% aqueous HBr or stirring with boron tribromide in dichloromethane. Alternatively, when the hydroxyl group is protected as a benzyl ester, the deprotection conditions may, for example, comprise hydrogenation with a palladium catalyst under a hydrogen atmosphere.

The preparation of all the above reactions and of the novel starting materials used in the aforementioned processes is conventional and the reaction conditions suitable for their performance or for the preparation and the process for isolating the desired product are known to the person skilled in the art by reference to the prior literature and to the examples and preparations herein.

Pharmaceutically acceptable salts of the compounds of formula (I) can be readily prepared by co-mixing a solution of the compound of formula (I) and a solution of the desired acid under suitable conditions. The salt may precipitate out of solution and be collected by filtration, or may be recovered by evaporation of the solvent.

Pharmaceutically acceptable salts of the compounds of formula (I) may be prepared by one or more of the following three methods:

(i) by reacting a compound of formula (I) with the desired acid;

(ii) by removing acid-or base-labile protecting groups from suitable precursors of compounds of formula (I) or by ring-opening of suitable cyclic precursors (e.g. caprolactone or caprolactam) using the desired acid; or

(iii) One salt of the compound of formula (I) is converted to the other by reaction with a suitable acid or by means of a suitable ion exchange column.

All three reactions are usually carried out in solution. The resulting salt may precipitate out and be collected by filtration or may be recovered by evaporation of the solvent. The degree of ionization of the resulting salt may vary from fully ionized to hardly ionized.

The compounds of formula (I) of the present invention have utility as MCR4 agonists in the treatment of a variety of disease states. Preferably, the MCR4 agonist exhibits functional potency (expressed as EC) at the MC4 receptor₅₀) Less than about 400nM, more preferably less than 200nM, still more preferably less than about 100nM, and more preferably still less than about 50nM, EC in which the functional potency of MCR4 can be performed using Protocol E as described in International patent application WO2005/077935₅₀And (6) measuring.

Combination therapy

The compounds of formula (I) or salts, solvates thereof of the present invention are delivered together with one or more other pharmaceutical agents. In addition, in certain instances, the compounds of formula (I) of the present invention, or salts, solvates, or prodrugs thereof, may be effectively delivered with an effective co-agent for reducing emesis. Some suitable pharmaceutical agents that may be used in conjunction with the present invention include:

1) compounds which modulate natriuretic factors, in particular atrial natriuretic factor (also known as atrial natriuretic peptide), B-and C-type natriuretic factors such as inhibitors or neuropeptide endonucleases, in particular compounds as described and claimed in WO 02/02513, WO 02/03995, WO 02/079143 and EP- ｃA-1258474, in particular the compound (2S) -2{ [1- {3-4 (-chlorophenyl) propyl ] amino } carbonyl) -cyclopentyl ] methyl } -4-methoxybutanoic acid of example 22 of WO 02/079143;

2) compounds which inhibit angiotensin converting enzyme, such as enalapril (enalapril), and combined inhibitors of angiotensin converting enzyme and endoneurin, such as omatralat (omapatrilat);

3) substrates for NO-synthetases, such as L-arginine;

4) cholesterol lowering agents, such as statins (e.g., atorvastatin/Lipitor)^TM) And fibrates (e.g., fenofibrate);

5) estrogen receptor modulators and/or estrogen agonists and/or estrogen antagonists, preferably raloxifene or lasofoxifene ((-) -cis-6-phenyl-5- [4- (2-pyrrolidin-1-yl-ethoxy) -phenyl ] -5, 6,7, 8-tetrahydronaphthalen-2-ol and pharmaceutically acceptable salts thereof, the preparation of which is described in detail in WO 96/21656);

6) PDE inhibitors, more specifically PDE2, 3, 4, 5,7 or 8 inhibitors, preferably PDE2 or PDE5 inhibitors, and most preferably PDE5 inhibitors (see below), preferably with an IC of less than 100nM for the respective enzyme₅₀Provided that the PDE 3 and 4 inhibitors are administered only topically or by injection into the penis to treat male erectile dysfunction);

7) vasoactive Intestinal Peptide (VIP), VIP mimetics, VIP analogues, more specifically those mediated by one or more VIP receptor subtypes VPAC1, VPAC or PACAP (pituitary adenylate cyclase activating peptide), one or more VIP receptor agonists or VIP analogues (e.g., Ro-125-1553) or VIP fragments, combinations of one or more alpha-adrenoreceptor antagonists with VIP (e.g., invigorp, Aviptadil);

8) serotonin receptor agonists, antagonists or modulators, more particularly agonists, antagonists or modulators of 5HT1A (including VML 670[ WO02/074288] and flibanserin [ US2003/0104980]) and/or agonists, antagonists or modulators of 5HT2A, 5HT2C, 5HT3 and/or 5HT6 receptors, including those described in WO-09902159, WO-00002550 and/or WO-00028993;

9) testosterone substitutes (including dehydroandrostenedione), testosterone (e.g. Tostrelle)^TM、LibiGel^TM) Dihydrotestosterone or a testosterone implant;

10) selective androgen receptor modulators, such as LGD-2226;

11) estrogen, estrogen and medroxyprogesterone acetate (MPA) (i.e., as a combination), or estrogen and methyltestosterone hormone replacement therapy agents (e.g., HRT, especially pimarin (Premarin), Cenestin, oesstrofemal, Equin, Estrace, norkunmide (Estrofem), eleste Solo, Estring, eastaderm TTS, eastaderm matrix, demerit (Dermestril), temphase, Preempro, Prempak, Premique, estetrest, Estratetest HS, Tibolone (Tibolone));

12) modulators of the norepinephrine, dopamine and/or serotonin transporters, such as bupropion (bupropion), GW-320659;

13) an agonist or modulator of the oxytocin/vasopressin receptor, preferably a selective oxytocin agonist or modulator;

14) dopamine receptor agonists or modulators, preferably D3 or D4 selective agonists or modulators. Such as apomorphine (apomorphine); and

15) antiemetic agents, e.g. 5-HT₃An antagonist or a neurokinin-1 (NK-1) antagonist.

Suitable 5-HT₃Antagonists include, but are not limited to, granisetron (granisetron), ondansetron (ondansetron), tropisetron (tropisetron), ramosetron (ramosetron), palonosetron (palonosetron), indisetron (indisetron), dolasetron (dolasetron), alosetron (aloetron), and azasetron (azasetron).

Suitable NK-1 antagonists include, but are not limited to, aprepitant, casopritant, epirubitant, cilapitant, netupitant, vettipitant, voroflitant, and 2- (R) - (1- (R) -3, 5-bis (trifluoromethyl) phenyl) ethoxy-4- (5- (dimethylamino) methyl-1, 2, 3-triazol-4-yl) methyl-3- (S) - (4-fluorophenyl) morpholine. See, for example, International patent application No. WO 2006/049933.

In particular, the use of the compounds of the invention for the treatment of lower urinary tract dysfunction in combination with other agents including, but not limited to:

muscarine acetylcholine receptor antagonists such as tolterodine;

an alpha-adrenergic receptor antagonist, in particular an alpha 1-adrenergic receptor antagonist or an alpha 2-adrenergic receptor antagonist;

an alpha-adrenergic receptor agonist or partial agonist, in particular an alpha 1-adrenergic receptor agonist or partial agonist or an alpha 2-adrenergic receptor agonist or partial agonist;

5HT2C agonists (see WO 2004/096196);

serotonin and Norepinephrine Reuptake Inhibitors (SNRI);

norepinephrine Reuptake Inhibitors (NRIs), such as reboxetine (in its racemic form or [ S, S ] -enantiomeric form);

capsaicin receptor [ VR ] antagonists, such as capsaicin;

α 2 δ ligands such as gabapentin (gabapentin) or pregabalin (pregabalin);

β 3 adrenergic receptor agonists;

5HT1a receptor antagonist or 5HT1a receptor inverse agonist;

prostaglandin receptor antagonists, such as EP1 receptor antagonists.

With respect to the use of compounds of formula (I) in the treatment of obesity and related disorders, the compounds may also be used in conjunction with other anti-obesity agents. Suitable anti-obesity agents include cannabinoid-1 (CB-1) receptor antagonists (such as rimonabant), apolipoprotein-B secretion/microsomal triglyceride transporter (apo-B/MTP) inhibitors (in particular, gut-selective MTP inhibitors such as ipapade or dirlotapede), 11 β inhibitor-inhibitors of hydroxysteroid dehydrogenase-1 (11 β -HSD type 1), peptide YY_3-36And mimetics thereof, cholecystokinin-A (CCK-A) agonists, monoamine reuptake inhibitors (such as sibutramine (sibutramine)), sympathomimetic agents,. beta.3 adrenergic receptor agonists, dopamine receptor agonists (such as bromocriptine (bromocriptine)), melanocyte stimulating hormone receptor mimetics, 5HT2c receptor agonists, melanin concentrating hormone antagonists, leptin (leptin) (OB protein), leptin mimetics, leptin receptor agonists, galanin (galanin) antagonists, lipase inhibitors (such as tetrahydrolipstatin (tetrahydrolipstatin), i.e., orlistat (orlistat)), anorectic drugs (such as bombesin (bomycin) agonists), neuropeptide-Y receptor antagonists (particularly, NPY-5 receptor antagonists), thyromimetics, dehydroepiglucocorticoids or mimetics thereof, androgen receptor agonists or antagonists, antagonists of the orexin receptor, agonists of the glucagon-like peptide-1 receptor, ciliary neurotrophic factors (such as Axokine)^TMCommercially available from Regeneron Pharmaceuticals, Inc., Tarrytown, NY and Procter&Gamble Company, Cincinnati, OH), human agony-related protein (AGRP) inhibitors, growth hormone releasing peptide (ghrelin) receptor antagonists, histamine 3 receptor antagonists or inverse agonists, neurointerleukin U receptor agonists, and the like. Other anti-obesity agents, including the preferred agents described hereinafter, are known to those skilled in the art or are readily apparent from the disclosure of the prior art. The compounds of the present invention may also be used in combination with natural compounds for lowering plasma cholesterol levels. Such natural compounds are commonly referred to as nutraceuticals and include, for example, garlic extract, Hoodia plant extract, and niacin. Preferred therefrom are anti-obesity agents selected from the group consisting of: CB-1 antagonists, gut-selective MTP inhibitors, orlistat, sibutramine, bromocriptine, ephedrine, leptin, peptide YY_3-36And mimetics thereof, and pseudoephedrine. Preferably, the compounds of the present invention and combination therapies for the treatment of obesity and related disorders are administered in conjunction with exercise and a healthy diet. Preferred CB-1 antagonists include rimonabant (SR141716A, also under its tradename Acomplia) described in U.S. Pat. No. 5,624,941^TMKnown from Sanofi-Synthelabo); and compounds described in the following documents: U.S. patents 5,747,524, 6,432,984 and 6,518,264; U.S. patent application publications US2004/0092520, US2004/0157839, US2004/0214855, and US 2004/0214838; U.S. patent application serial No. 10/971599 filed on month 10, 22 of 2004; and PCT patent publications WO 02/076949, WO 03/075660, WO04/048317, WO04/013120, and WO 04/012671. Preferred gut-selective MTP inhibitors include diropernide as described in us patent 6,720,351; 4- (4- (4- (4- ((2- ((4-methyl-4H-1, 2, 4-triazol-3-ylthio) methyl) -2- (4-chlorophenyl) -1, 3-dioxolan-4-yl) methoxy) phenyl) pyridazin-1-yl) phenyl) -2-sec-butyl-2H-1, 2, 4-triazol-3 (4H) -one described in U.S. Pat. nos. 5,521,186 and 5,929,075 (R103757); and inpitapide (implitapide) as described in us patent 6,265,431 (BAY 13-9952). Other representative anti-obesity agents for use in the combinations, pharmaceutical compositions and methods of the invention may be prepared using methods known to those skilled in the art, for example: sibutramine can be prepared as described in U.S. patent No. 4,929,629; bromocriptine can be prepared as described in U.S. patents 3,752,814 and 3,752,888; orlistat can be prepared as described in U.S. patents 5,274,143, 5,420,305, 5,540,917 and 5,643,874; and PYY_3-36(including mimetics) can be prepared as described in U.S. patent application publication 2002/0141985 and WO 03/027637.

A preferred group herein is the combination of a compound of the invention with one or more other therapeutic agents selected from: a PDE5 inhibitor; an NEP inhibitor; a D3 or D4 selective agonist or modulator; an estrogen receptor modulator and/or an estrogen agonist and/or an estrogen antagonist; testosterone replacement agents, testosterone or testosterone implants; estrogen, estrogen with medroxyprogesterone or medroxyprogesterone acetate (MPA), or estrogen with methyltestosterone hormone replacement therapy.

A preferred combination for the treatment of MED is a combination of a compound of the invention and one or more PDE5 inhibitors and/or NEP inhibitors.

A preferred combination for the treatment of FSD is a combination of a compound of the invention with: PDE5 inhibitors and/or 5HT1a receptor antagonists and/or NEP inhibitors and/or D3 or D4 selective agonists or modulators and/or estrogen receptor modulators, estrogen agonists, estrogen antagonists and/or testosterone substitutes, testosterone implants and/or estrogen, estrogen and medroxyprogesterone or medroxyprogesterone acetate (MPA), estrogen and methyltestosterone hormone replacement therapeutics.

Particularly preferred PDE5 inhibitors for use in such a combination product for the treatment of MED or FSD are selected from: 5- [ 2-ethoxy-5- (4-methyl-1-piperazinylsulfonyl) phenyl ] -1-methyl-3-n-propyl-1, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one (sildenafil), particularly present as the citrate salt);

(6R, 12aR) -2, 3, 6,7, 12, 12 a-hexahydro-2-methyl-6- (3, 4-methylenedioxyphenyl) -pyrazino [2 ', 1': 6, 1] pyrido [3, 4-b ] indole-1, 4-dione (IC-351 or tadalafil);

2- [ 2-ethoxy-5- (4-ethyl-pyridazin-1-yl-1-sulfonyl) -phenyl ] -5-methyl-7-propyl-3H-imidazo [5, 1-f ] [1,2,4] triazin-4-one (vardenafil));

5- (5-acetyl-2-butoxy-3-pyridinyl) -3-ethyl-2- (1-ethyl-3-azetidinyl) -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one;

5- (5-acetyl-2-propoxy-3-pyridyl) -3-ethyl-2- (1-isopropyl-3-azetidinyl) -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one;

5- [ 2-ethoxy-5- (4-ethylpyridazin-1-ylsulfonyl) pyridin-3-yl ] -3-ethyl-2- [ 2-methoxyethyl ] -2, 6-dihydro-7H-pyrazolo [4, 3-d ] pyrimidin-7-one;

4- [ (3-chloro-4-methoxybenzyl) amino ] -2- [ (2S) -2- (hydroxymethyl) pyrrolidin-1-yl ] -N- (pyrimidin-2-ylmethyl) pyrimidine-5-carboxamide (avanafil);

3- (1-methyl-7-oxo-3-propyl-6, 7-dihydro-1H-pyrazolo [4, 3-d ] pyrimidin-5-yl) -N- [2- (1-methylpyrrolidin-2-yl) ethyl ] -4-propoxybenzenesulfonamide (udenafil);

7- (3-bromo-4-methoxy-benzyl) -1-ethyl-8- (2-hydroxy-cyclopentylamino) -3- (2-hydroxy-ethyl) -3, 7-dihydro-purine-2, 6-dione (dasatinafil);

and pharmaceutically acceptable salts thereof.

Particularly preferred NEP inhibitors for use in such a combination product for the treatment of MED or FSD are the compounds exemplified in WO 02/079143.

By reference to the compounds contained in the patents and patent applications which can be used according to the present invention, we mean therapeutically active compounds as defined in the claims (in particular claim 1) and the specific examples (all of these documents are incorporated herein by reference).

If a combination of active agents is administered, they may be administered simultaneously, separately and sequentially in the same or different formulations.

Physiological experiments

Melanocortin receptor agonist activity, selectivity

Compounds directed against melanocortin receptors type 1 and 3 (MC1 and MC3) In vitro agonist potency (EC) ₅₀ ) The measurement of (2).

Activation of the Melanocortin (MC) receptor by agonists results in activation of intracellular adenylate cyclase, which synthesizes the second messenger signal molecule adenosine 3 ', 5' -cyclic monophosphate (cAMP). Changes in cAMP levels following treatment of recombinant MC1 and MC3 cell lines with test compounds were measured and MC1 and MC3 potency Estimates (EC) were calculated as follows₅₀)：

Using standard molecular biologyMethods Human Embryonic Kidney (HEK) or chinese hamster ovary cell lines stably transfected with full-length cDNA encoding the MC1 or MC3 receptors, respectively, were established. Test compounds were dissolved in dimethyl sulfoxide (DMSO) at 4 mM. Test compounds were prepared in 11-point semilog unit incremental dilution series, typically starting at 50uM, in a buffer consisting of: phosphate Buffered Saline (PBS), 2.5% DMSO, and 0.05% pluronic F-127 surfactant. 80-90% of the confluent freshly cultured cells were harvested and resuspended in Darber's Modified Eagle's Medium (DMEM). Cells (10,000 for MC3 and 20,000 for MC 1) were added to test compound dilution series in 384-well assay plates and incubated at 37 ℃ for 1 hour. The relevant cAMP concentration in each well was then measured using the β -galactosidase fragment complementation method purchased as a kit, discover cAMP II kit from GE Healthcare/Amersham biosciences uk. In the case of MC1, 3-isobutyl-1-methylxanthine (IBMX) was included at a concentration of 750. mu.M in DMEM when the cells were resuspended for analysis. The fluorescence readings taken from each assay well were converted to% effect relative to the maximum value of the control well corresponding to the concentration of melanocyte stimulating hormone demonstrated to provide the greatest effect. Fitting sigmoid curves to log using a custom software application named SIGHTS₁₀Point of Effect of inhibitor concentration on% EC as test Compound concentration determined by software₅₀The estimate gives the half-way effect (effect half way) between the bottom and top asymptotes of the sigmoidal dose response curve. Each experiment contained EC on alpha-melanocyte-stimulating hormone₅₀Determination of the data used to show experimental consistency and to allow EC to be obtained in different experiments₅₀A criterion for making a fair comparison between the estimates.

EC of MC5 and MC4 by the methods described in Protocols D and E of US2005/0176772 (pages 28 to 30), respectively₅₀The activity was measured.

Nle4, D-Phe 7-alpha-MSH inhibition of the MC4 receptor

Nle4，D-Phe7-αMSH is a stable analogue of Melanocyte Stimulating Hormone (MSH), which is an agonist of the MC4 receptor (MC 4R). Can use para 2¹²⁵I]Competitive binding experiments with Nle4, D-Phe7- α -MSH Compounds were tested for their ability to inhibit the membrane binding of Nle4, D-Phe7- α -MSH to MC4R expressing cells.

Cells expressing MC4R were subjected to homogenization and membrane debris was separated by differential centrifugation. CHO-CRE MC4R cell membranes were coupled to PVT-PEI-WGA SPA bead type A for 2 hours, spun at 1000RPM for 5min and suspended to a concentration of 300. mu.g beads/ml (0.15. mu.g membrane, 15. mu.g beads/well). Total volume 50. mu.l buffer (25mM HEPES, 1mM MgCl) per well₂，2.5mM CaCl₂1% Pluronic F68, 1 complete EDTA protease inhibitor tablet/50 ml pH7) with 0.06nM [ 2]¹²⁵I]The bead/membrane mixture was incubated in duplicate with Nle4, D-Phe7- α -MSH and 11.5 log concentration of competing ligand. Nonspecific binding was determined by the addition of 100nM SHU 9119. The reaction was initiated by adding beads/membrane and the plate was incubated at room temperature for 12 hours (first hour on a plate shaker) and then the amount of radioactivity present was determined using a Wallac plate counter. Data analysis was performed using appropriate software to determine Ki values.

Nle4, D-Phe 7-alpha-MSH inhibition of the MC3 receptor

Nle4, D-Phe 7-alpha-MSH is a stable analog of Melanocyte Stimulating Hormone (MSH), which is an agonist of the MC3 receptor (MC 3R). Can use para 2¹²⁵I]Competitive binding experiments with Nle4, D-Phe7- α -MSH Compounds were tested for their ability to inhibit the membrane binding of Nle4, D-Phe7- α -MSH to MC3R expressing cells.

Cells expressing MC3R were subjected to homogenization and membrane debris was separated by differential centrifugation. CHO-CRE MC3R cell membranes were coupled to PVT-PEI-WGA SPA bead type A for 2 hours, spun at 1000RPM for 5min, and suspended to a final test concentration of 800. mu.g beads/ml (1.2. mu.g membrane, 40. mu.g beads/well). Total volume in 50. mu.l buffer (25mM HEPES, 1mM MgCl) per well₂，2.5mM CaCl₂1% Pluronic F68, 1 complete EDTA protease inhibitor tablet/50 ml pH7) with 0.06nM [ 2]¹²⁵I]The bead/membrane mixture was incubated in duplicate with Nle4, D-Phe7- α -MSH and 11 semilog concentrations of competing ligand. Nonspecific binding was determined by the addition of 100nM SHU 9119. The reaction was initiated by adding beads/membrane and the plate was incubated at room temperature for 12 hours (first hour on a plate shaker) and then the amount of radioactivity present was determined using a Wallac plate counter. Data analysis was performed using appropriate software to determine Ki values.

High Density drug-drug interaction (DDI)3 μ M cocktail Screen

Drug interactions are situations where a substance affects the activity of another drug, i.e. the effect is enhanced or diminished or they together produce a new effect that the drug itself cannot produce. Drug interactions can be the result of different processes, but one that is relatively common is that one drug affects the pharmacokinetics of another drug by inhibiting the cytochrome P450 that metabolizes the other drug. Because of the importance of these phenomena, the evaluation of the DDI potential of Novel Chemical Entities (NCE) is considered to be very important early in the drug development process.

DDI cocktail screening (HLM) in human liver microsomes was performed in a fully automated mode and the purpose of the screening was to provide a single point assessment of the DDI potential of the 4 major cytochrome P450 enzymes 1a2, 2D6, 2C9 and 3a4 by a novel chemical entity.

The substrate cocktail method of P450DDI utilizes human liver microsomes along with clinical drug probes specific for the isoform and allows simultaneous measurement of the known activity of P4501a2, 2C9, 2D6 and 3a4 in a single incubation. The method is performed at high throughput and metabolite detection is performed simultaneously by LC-MS/MS. This method has been thoroughly tested and evaluated using standard compounds.

Source of microsomes	Pooled human liver microsomes
		Concentration of microsomes	0.1mg/mL

P450 concentration	0.03μM
		Regeneration system	NADPH(1.3mM)
Time of detection	8min
Probe substrate (probed enzyme)	Concentration of
		9-Aminotetrahydroacridine (Tacrine) (1A2)	2μM
Difluorofenac (Diclofenac) (2C9)	5μM
		Dextromethorphan (Dextromeorphan) (2D6)	5μM
Midazolam (Midazolam) (3A4)	2μM
Inhibitors	Concentration of
		NCE (test Compound)	3μM
Miconazole (Miconazole) (general purpose control)	3μM

The expression of metabolites of each substrate was examined over time in the presence and absence of NCE (test compound/inhibitor) at a concentration of 3. mu.M. The inhibitory potential of the compounds was assessed (as percentage values) and illustrated using the following protocol. These data are then used in conjunction with other measurements to evaluate the suitability of the NCE and to aid in the design and development of drugs.

% inhibition	IC50
		＞75％	＜1μM
25-75％	1-10μM
		＜25％	＞10μM

In vitro metabolic rate determination (human liver microsome (HLM); Rat Liver Microsome (RLM) assay)

Many drugs are metabolized by the cytochrome P450 monooxygenase system. This enzyme is found in high concentrations in the liver and binds to the endoplasmic reticulum of hepatocytes. Such an enzyme system can be obtained in a semi-purified state by preparing a liver microsomal fraction. In such systems the in vitro half-life of the compound is determined to provide a useful indication of metabolic stability.

Materials and reagents

All reagents were of analytical grade.

1.200mM phosphate buffer (Sigma) -100ml of 1M phosphate buffer pH 7.4 was dissolved in 400ml of ultrapure water. The pH is adjusted to 7.4 using concentrated orthophosphoric acid, if necessary, prepared monthly and stored under refrigeration (2-8 ℃).

2.0.1M MgCl₂.6H₂O (BDH) -2.032g was dissolved in 100ml of ultrapure water and stored under refrigeration (2-8 ℃ C.).

3.0.02M NADP (Sigma) -15.3mg were dissolved in 1000. mu.l of ultrapure water-then stored refrigerated (2-8 ℃) for further use.

4.0.1M D-L Isocitric acid (Sigma) -129mg were dissolved in 5ml ultrapure water-then stored refrigerated (2-8 ℃) for further use.

5. Isocitrate dehydrogenase, form IV (Sigma) -stored refrigerated (2-8 ℃ C.).

6. Substrate stock solutions (about 1mg/ml) in miscible organic solvents (such as methanol, ethanol) or water, stored refrigerated (2-8 ℃).

7.50mM p-nitroanisole (PNA) (Aldrich) -7.65mg dissolved in 1ml methanol, stored refrigerated (2-8 ℃) until ready to use.

8.50 μ M p-nitrophenol (PNP) (Sigma) -0.69mg in 100ml water and stored refrigerated (2-8 ℃ C.).

9.20% trichloroacetic acid (TCA) (BDH) -20g was dissolved in 100ml ultrapure water, prepared in amber glassware and stored at room temperature.

10.10M sodium hydroxide (BDH) -40g was dissolved in 100ml of ultrapure water (care should be taken when preparing this solution)Care is takenBecause the reaction is exothermic), prepared in "break-safe" glassware and stored at room temperature.

11. Liver microparticles or Supermix microparticles stored at-80 ℃, which should be used immediately after thawing, kept on ice and dispensed.

12. Ultrapure water.

13. A thermostatically controlled vibrating water bath set to provide a temperature of about 37 ℃ during incubation.

14. Reagents for terminating the incubation (usually organic solvents, acids or bases).

Method for in vitro metabolic rate determination using liver microsomes or Supermix microsomes

The method described below was used for an incubation volume of 1.5 ml.

1. The following mixtures were prepared in test tubes:

reagent

Stock solution concentration

Incubation concentration

Volume of addition

			(for 1.5ml incubation)
				Phosphate buffer pH 7.4	200mM	50mM	375ul
MgCl2	0.1M	5mM	75ul
				Isocitric acid	0.1M	5mM	75ul
Isocitrate dehydrogenase	See above the bottle	1 unit/ml	See below

Calculating the volume for each new batch of isocitrate dehydrogenase

For example, the protein concentration is 18mg/ml

Enzyme activity 3.3 units/mg

Thus the specific activity was 3.3 × 18 units/ml to 59 units/ml

For an incubation of 1.5ml,

2. the microsomes were thawed at room temperature and sufficient microsomes were added to give an incubation with a final concentration of 0.5nmol cytochrome P450/ml, e.g., for 1.5ml incubation. The volume of microsomes added is required to be:

3. sufficient ultrapure water was added to give a total incubation volume of 1.425 ml.

4. 237.5. mu.l of the incubation mixture was taken and placed in a tube as PNA positive control. Add 2.5. mu.l of PNA solution, vortex mix, and place the tube into the rack of a thermostatically controlled vibrating water bath.

5. 100 μ l was taken as a no substrate control and dispensed into tubes. The test tubes were placed in racks in a thermostatically controlled vibrating water bath.

6. Substrate was added to the incubation. The substrate should have an initial concentration of 1. mu.M. The substrate required for the remaining 1.162.5ml incubations was calculated as follows:

note: the volume of organic solvent added should not exceed 0.1% of the total incubation volume.

7. 100 μ l of the incubation mixture was added to the tube as a control without cofactor. Vortex mixed and placed into a rack of thermostatically controlled vibrating water bath.

8. The tubes containing the incubation mixture, as well as the positive control and no cofactor, were placed in a rack set to a thermostatically controlled vibrating water bath at 37 ℃ for a pre-incubation of about 5 min.

9. NADP was added to start the reaction (75. mu.l for 1.162.5ml incubation mixture, 12.5. mu.l for positive control tube, and 5. mu.l for no substrate tube) and the first time point was obtained. PNA positive controls, no cofactor controls and no substrate tubes were incubated for the full incubation time.

10. 100 μ l aliquots were taken at up to 9 different sampling times (typically 0,3, 5, 10, 15, 20,30, 45 and 60min) from 0 to 60min and the reaction was terminated. Longer incubations may be used, but after 120min the microsomes deteriorate. The reaction can be terminated by the addition of an organic solvent, acid or base. The no cofactor control and the no substrate control were treated in a similar manner at the end of the incubation process, i.e., terminated with the same reagents.

11.PNA positive control program：

After the last sample had been taken, a positive control was taken and 1ml 20% TCA was added to the tube. A tube containing 250. mu.l of 50. mu.M PNP standard was also prepared and 1ml of 20% TCA was added. Both tubes were vortexed and allowed to sit for about 5min to precipitate the protein.

Both tubes were centrifuged for about 5min in an apparatus set at 3500 rpm. 1ml of the supernatant was taken and placed in a clean tube, the remainder being discarded.

1ml of 10M NaOH was added to the supernatant, mixed by vortexing, and allowed to stand for about 5 min. The absorbance of the PNP standard against distilled water was then measured at 400nm with distilled water as a blank spectrophotometer. Microsomal 4-nitrobenzyl ether O-demethylase activity was calculated as follows:

calculation of results

Absorbance samples x PNP in nmol standard (i.e. 12.5nmol)

Absorption rate PNP standard substance x 60 x 0.125

＝nmol/min/nmol P450

The activity value of the incubation must be equal to or greater than 85% of the mean value of the batches used for effective incubation. If this criterion is not met, the incubation must be repeated.

11. Samples were analyzed by substrate specific assays (including no cofactor control and no substrate control) to determine disappearance kinetics (dispeparrance kinetics).

Data analysis

The data obtained using the above method can be quantified in terms of intrinsic clearance of the substrate in vitro (Clint). Assuming substrate concentrations below Km, metabolism should be first order and give a log-linear curve of substrate disappearance over time.

The in vitro half-life of the substrate can be determined by: measurements of relative substrate concentrations (e.g., drug/internal standard ratio) are plotted over time and a line that best fits the data is fitted. The gradient of the line is the first order rate constant (k) of substrate disappearance and is determined by regression analysis. This rate constant can be converted to a half-life according to the following equation.

Alternatively, the rate constant can be converted to intrinsic clearance (Clint) by the following equation.

Clint (μ l/min/mg) ═ k/protein concentration in incubation (mg/ml)). 1000

Application method

The compounds of the invention intended for pharmaceutical use may be administered as crystalline or amorphous products. They can be obtained, for example, as suppositories, powders or films by methods such as precipitation, crystallization, freeze-drying, spray-drying or evaporation-drying. Microwave or high frequency drying may also be used for this purpose.

They may be administered alone or in combination with one or other mixtures of the present invention or one or more other drugs (or as any combination thereof). Generally, they can be administered as a formulation in combination with one or more pharmaceutically acceptable excipients. The term "excipient" is used herein to describe any ingredient other than a compound of the present invention. The choice of excipient will depend in large part on factors such as the particular mode of administration, the effect of the excipient on solubility and stability, and the nature of the dosage form.

Pharmaceutical compositions suitable for delivery of the compounds of the invention and methods for their preparation will be apparent to those skilled in the art. Such compositions and methods for their preparation can be found, for example, in the following documents:Remington’s Pharmaceutical Sciences19 th edition (mack publishing Company, 1995).

Accordingly, the present invention provides a pharmaceutical composition comprising a compound of formula (I) in association with a pharmaceutically acceptable diluent or carrier.

Any suitable route of administration may be employed to provide an effective dose of a compound of the invention to a mammal, especially a human. For example, oral administration (including buccal and sublingual administration), rectal administration, topical administration, parenteral administration, ocular administration, pulmonary administration, nasal administration, and the like may be administered. Dosage forms include tablets, lozenges, dispersions, suspensions, liquids, capsules, creams/ointments, aerosols, and the like. Preferably, the compound of formula (I) is administered orally or nasally.

The effective dose of the active ingredient may vary depending on the following factors: the particular compound used, the mode of administration, the characteristics of the mammal to be treated (e.g., body weight), the disease state to be treated, and the severity of the disease state to be treated. The dosage can be readily determined by one skilled in the art.

For the treatment of sexual dysfunction, the compounds of the present invention are administered in the following dosage ranges: from about 0.001 milligram (mg) to about 1000mg, preferably from about 0.001mg to 500mg, more preferably from about 0.001mg to 100mg, even more preferably from about 0.001mg to 50mg, and particularly preferably from about 0.002mg to 25mg per kilogram of body weight, preferably as a single dose administered orally or as a nasal spray. For example, oral administration may require a total dose of about 0.1mg up to about 1000mg, whereas intravenous doses may require only about 0.001mg up to about 100 mg. The total daily dose may be administered in a single dose or in divided doses, or may fall outside the general ranges given herein, as directed by the physician.

When treating obesity with diabetes and/or hyperglycemia or obesity alone, satisfactory results are generally obtained when the compounds of the invention are administered in the following daily doses: the daily dose is from about 0.0001mg to about 1000mg, preferably from about 0.001mg to about 500mg, more preferably from about 0.005mg to about 100mg, especially preferably from about 0.005mg to about 50mg, administered per kg of animal body weight, preferably in a single dose or in divided doses from 2 to 6 times daily or in sustained release form. In the case of a 70kg adult, the total daily dose will generally be from about 0.7mg up to about 3500 mg. The dosage regimen may be adjusted to provide the optimal therapeutic response.

When treating diabetes and/or hyperglycemia, as well as diseases or conditions for which compounds of formula (I) are useful, satisfactory results are generally obtained when about 0.001mg to about 100mg of a compound of the invention is administered per kilogram of animal body weight, preferably in a single dose or in divided doses from 2 to 6 times daily, or in sustained release form. In the case of a 70kg adult, the total daily dose will generally be from about 0.07mg up to about 350 mg. The dosage regimen may be adjusted to provide the optimal therapeutic response.

These doses are based on an average human subject weighing about 65kg to 70 kg. A physician will be able to readily determine dosages for subjects with weights outside this range, such as infants, elderly and obese subjects.

The compounds of the invention may be administered orally. Oral administration may include swallowing to allow the compound to enter the gastrointestinal tract and/or buccal, lingual or sublingual administration whereby the compound enters the blood stream directly from the mouth.

Formulations suitable for oral administration include solid, semi-solid and liquid systems, such as tablets; soft or hard capsules comprising multiparticulates or nanoparticles, liquids or powders; lozenges (including liquid filled); a masticatory; gelling agent; a fast-dispersing dosage form; a thin film agent; ovule agents (ovules); a spray; and buccal/mucoadhesive patches.

Liquid preparations include suspensions, solutions, syrups and elixirs. Such formulations may be employed as fillers for soft or hard capsules (made, for example, from gelatin or hydroxypropylmethyl cellulose) and typically contain a carrier, such as water, ethanol, polyethylene glycol, propylene glycol, methylcellulose, or a suitable oil, and one or more emulsifying agents and/or suspending agents. Liquid formulations may also be prepared by reconstitution of a solid, for example, from a pharmaceutical pack.

The compounds of the invention may also be administered in combination with other therapeutic agentsFast dissolving, fast disintegrating dosage forms, such as those described in the following references: the Expert opinions in Therapeutic documents,11(6)，981-986 by Liang and Chen(2001)。

for a tablet dosage form, depending on the dosage, the drug may constitute from 1 wt% to 80 wt% of the dosage form, more typically from 5 wt% to 60 wt% of the dosage form. In addition to the drug, tablets typically contain a disintegrant. Examples of disintegrants include sodium starch glycolate, sodium carboxymethylcellulose, calcium carboxymethylcellulose, croscarmellose sodium, crospovidone, polyvinylpyrrolidone, methylcellulose, microcrystalline cellulose, lower alkyl-substituted hydroxypropylcellulose, starch, pregelatinized starch, and sodium alginate. Typically, the disintegrant will comprise from 1 wt% to 25 wt% of the dosage form, preferably from 5 wt% to 20 wt% of the dosage form.

Binders are commonly used to impart cohesive properties to the tablet formulation. Suitable binders include microcrystalline cellulose, gelatin, sugars, polyethylene glycol, natural and synthetic gums, polyvinylpyrrolidone, pregelatinized starch, hydroxypropyl cellulose, and hydroxypropyl methyl cellulose. Tablets may also contain diluents such as lactose (monohydrate, spray-dried monohydrate, anhydrous, etc.), mannitol, xylitol, dextrose, sucrose, sorbitol, microcrystalline cellulose, starch, and dicalcium phosphate dihydrate.

The tablets may also optionally contain surfactants such as sodium lauryl sulfate and tween 80, and glidants such as silicon dioxide and talc. When present, the surfactant may comprise 0.2 wt% to 5 wt% of the tablet, and the glidant may comprise 0.2 wt% to 1 wt% of the tablet.

Tablets also typically contain lubricating agents such as magnesium stearate, calcium stearate, zinc stearate, sodium stearyl fumarate, and mixtures of magnesium stearate and sodium lauryl sulfate. The lubricant will generally comprise from 0.25 wt% to 10 wt%, preferably from 0.5 wt% to 3 wt% of the tablet.

Other possible ingredients include antioxidants, coloring agents, flavoring agents, preservatives, and taste masking agents.

Exemplary tablets comprise up to about 80% drug, about 10 wt% to about 90 wt% binder, about 0 wt% to 85 wt% diluent, about 2 wt% to about 10 wt% disintegrant, and about 0.25 wt% to about 10 wt% lubricant.

The tablet blend may be compressed into tablets either directly or with rollers. Tablet blends or partial blends may be wet, dry or melt granulated, melt refrigerated or extruded prior to tableting. The final formulation may comprise one or more layers, and may be coated or uncoated; it may even be encapsulated.

The formulation of tablets is discussed in the following documents: of H.Lieberman and L.LachmanPharmaceutical Dosage Forms：TabletsVol.1 (Marcel Dekker, New York, 1980).

Consumable oral films for human or veterinary use are generally flexible, water-soluble or water-swellable film dosage forms which dissolve rapidly or may be mucoadhesive and generally comprise a compound of formula (I), a film-forming polymer, a binder, a solvent, a humectant, a plasticizer, a stabilizer or emulsifier, a viscosity modifier and a solvent. Some components of the formulation may perform more than one function.

The compounds of formula (I) may be water soluble or water insoluble. The water soluble compound typically comprises from 1 wt% to 80 wt%, more typically from 20 wt% to 50 wt% of the solute. Less soluble compounds may comprise a greater fraction of the composition, typically up to 88 wt% of the solute. Alternatively, the compound of formula (I) may be in the form of multiparticulate beads.

The film-forming polymer may be selected from natural polysaccharides, proteins or synthetic hydrocolloids and is typically present in the range of 0.01 to 99 wt%, more typically in the range of 30 to 80 wt%.

Other possible ingredients include antioxidants, colorants, flavors and fragrances, preservatives, saliva stimulating agents, cooling agents, co-solvents (including oils), emollients, fillers, anti-foaming agents, surfactants and taste masking agents.

The thin film according to the invention is typically prepared by evaporation drying of an aqueous film coated onto a releasable backing support or paper. This can be done in a drying oven or drying tunnel, typically a combination coating dryer, or can be done by freeze drying or vacuum.

Solid formulations for oral administration may be formulated for immediate release and/or modified release. Modified release dosage forms include delayed release, sustained release, pulsed release, controlled release, targeted release and programmed release.

Modified release formulations for the purposes of the present invention are described in U.S. Pat. No. 6,106,864. Details of other suitable delivery techniques (such as high energy dispersion and isotonic and coating particles) may be found in Verma et alPharmaceutical Technology On-line25(2), 1-14 (2001). The use of a chew for achieving controlled release is described in WO 00/35298.

The compounds of the invention may also be administered directly into the bloodstream, into muscle or into internal organs. Suitable parenteral administration modes include intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurethral, intrasternal, intracranial, intramuscular, intrasynovial and subcutaneous administration. Suitable means for parenteral administration include needle (including microneedle) syringes, needle-free syringes and infusion techniques.

Parenteral formulations are typically aqueous solutions which may contain excipients such as salts, sugars and buffers (preferably to a pH of 3 to 9), but for some applications may be more suitably formulated as sterile non-aqueous solutions or as dry forms for administration in conjunction with a suitable vehicle such as sterile, pyrogen-free water.

Preparation of parenteral formulations under sterile conditions can be accomplished using standard pharmaceutical techniques well known to those skilled in the art, for example, by freeze-drying.

The solubility of the compound of formula (I) for use in preparing parenteral solutions can be increased by using suitable formulation techniques, such as the addition of a solubilizing agent.

Formulations for parenteral administration may be formulated for direct release and/or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release and programmed release. The compounds of the present invention may thus be formulated as suspending agents or as solid, semi-solid or thixotropic liquids for administration as an implanted reservoir providing modified release of the active substance. Examples of such formulations include drug-coated stents (stents) and semi-solids and suspensions containing drug-loaded poly (dl-lactic-co-glycolic acid) (PGLA) microbeads.

The compounds of the invention may also be administered topically, intradermally (intra) or transdermally to the skin or mucosa. Typical formulations for this purpose include gels, hydrocolloids, lotions, solutions, creams, ointments, dusting powders, dressings, foams, films, patches, wafers, implants, sponges, fibers, bandages and microemulsions. Liposomes may also be used. Typical carriers include alcohols, water, mineral oil, liquid paraffin, white petrolatum, glycerin, polyethylene glycol, and propylene glycol. Penetration enhancers may be added-see, for example, J Pharm Sci by Finnin and Morgan,88(10) 955- "958 (10 months 1999).

Other means of topical application include by electroporation, iontophoresis, sonophoresis, and microneedle or needle-free (e.g., Powderject)^TM、Bioject^TMEtc.) injection delivery.

Formulations for topical administration may be formulated for direct release and/or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release and programmed release.

The compounds of the invention may also be administered intranasally or by inhalation, typically in the form of a dry powder from a dry powder inhaler (alone, as a mixture (e.g. a dry blend with lactose), or as a mixed component particle (e.g. mixed with a phospholipid such as lecithin), or as an aerosol from a pressurized container, pump, jet, nebuliser (preferably one using electrohydrodynamic forces to produce a mist) or nebuliser (with or without the use of a suitable propellant such as 1,1, 1, 2-tetrafluoroethane or 1,1, 1,2, 3,3, 3-hexafluoropropane) or as nasal drops.

The pressure vessel, pump, sprayer, atomizer or nebulizer contains a solution or suspension of a compound of the invention, including, for example, ethanol, aqueous ethanol or other suitable agent for dispersing, dissolving or prolonging the release of the active, a propellant as a solvent, and optionally a surfactant (such as sorbitan trioleate, oleic acid or oligolactic acid).

Prior to use in dry powder or suspension formulations, the drug product is micronized to a size suitable for delivery by inhalation (typically less than 5 microns). This may be achieved by any suitable comminution method, such as spiral jet comminution, fluidized bed jet comminution, supercritical fluid processing to form nanoparticles, high pressure homogenization or spray drying.

Capsules (made, for example, from gelatin or hydroxypropylmethyl cellulose), vesicles and cartridges for use in an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention, a suitable powder base such as lactose or starch, and a performance modifier such as l-leucine, mannitol or magnesium stearate. Lactose may be anhydrous or may be in the monohydrate form, the latter being preferred. Other suitable excipients include dextrose, glucose, maltose, sorbitol, xylitol, fructose, sucrose, and trehalose.

Solutions suitable for use in nebulizers that use electrohydrodynamic forces to generate a mist may contain 1 μ g to 20mg of a compound of the invention per actuation, and the actuation volume may vary from 1 μ l to 100 μ l. A typical formulation may comprise a compound of formula (I), propylene glycol, sterile water, ethanol and sodium chloride. Alternative solvents that may be used in place of propylene glycol include glycerol and polyethylene glycol.

Flavoring agents (such as menthol and levomenthol) or sweetening agents (such as saccharin or saccharin sodium) may be added to those formulations of the invention intended for inhalation/intranasal administration.

Formulations for topical administration may be formulated for direct release and/or modified release using, for example, PGLA. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release and programmed release.

For dry powder inhalers and aerosols, the dosage unit is determined by delivering a metered amount of the valve. Units according to the invention are generally configured to administer a metered amount or "puff" comprising from 0.001mg to 10mg of a compound of formula (I). The total daily dose will generally be in the range of from 0.001mg to 40mg, which may be administered in a single dose or more usually in divided doses throughout the day.

The compounds of the invention may be administered rectally or vaginally, for example in the form of suppositories, pessaries or enemas. Cocoa butter is a traditional suppository base, but many alternatives can be used as desired.

Formulations for rectal/vaginal administration may be formulated for direct release and/or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release and programmed release.

The compounds of the invention may also be administered directly to the eye or ear, usually in the form of drops of micronized suspension or solution in isotonic, pH adjusted, sterile saline. Other formulations suitable for ocular or otic administration include ointments, gels, biodegradable implants (e.g., absorbable gel sponges, collagen) and non-biodegradable implants (e.g., silicone), wafers, lenses, and particulate or vesicular systems such as niosomes or liposomes. The following polymers may be added with preservatives such as benzalkonium chloride: such as crosslinked polyacrylic acid, polyvinyl alcohol, hyaluronic acid, cellulosic polymers (e.g., hydroxypropylmethylcellulose, hydroxyethylcellulose, or methylcellulose), or heteropolysaccharide polymers.

Formulations for ocular/otic administration may be formulated for direct release and/or modified release. Modified release formulations include delayed release, sustained release, pulsed release, controlled release, targeted release and programmed release. The compounds of the invention may be used in combination with suitable macromolecular entities such as cyclodextrins and derivatives thereof or polyethylene glycol containing polymers in any of the above forms of administration to improve solubility, dissolution rate, taste masking, bioavailability and/or stability thereof.

For example, drug-cyclodextrin complexes are generally found useful in most forms of administration routes. Both inclusion and non-inclusion complexes may be used. As an alternative to direct complexation with the drug, the cyclodextrin may be used as an auxiliary additive, i.e. as a carrier, diluent or solubiliser. The most frequently used for this purpose are alpha-, beta-and gamma-cyclodextrins, examples of which can be found in international patent applications WO 91/11172, WO 94/02518 and WO 98/55148.

Since the administration of a combination of active substances may be required for the purpose of treating a particular disease or disease state, the following are also included within the scope of the present invention: two or more pharmaceutical compositions, at least one of which comprises a compound of the invention, are conveniently combined in a kit suitable for co-administration of the compositions.

The kit of the invention thus comprises two more separate pharmaceutical compositions, at least one of which comprises a compound of the invention, and means for separately storing said compounds, such as a container, a separate bottle or a separate foil pouch. Examples of such kits are blister packs like the pack tablets, capsules etc.

The kit of the invention is particularly suitable for administration of different dosage forms, such as oral and parenteral dosage forms; separate compositions for administration at different dosage intervals; or for titration of different compounds with each other. To aid compliance, the kit typically includes directions for administration, and may be provided with so-called memory aids.

The invention is illustrated by the following non-limiting examples, in which the following abbreviations and definitions are used:

examples

Examples 1-57 were prepared according to scheme 1.

Example 1: 6- [ (3S,4R) -3- { [ 4-acetyl-5- (4-methoxyphenyl) -1, 4-diazacyclooctane- 1-radical]Carbonyl } -4- (2, 4-difluorophenyl) pyrrolidin-1-yl]Pyridazine-3-carbonitriles

To a solution of preparation 77 (45mg, 0.084mmol) in DCM (5mL) was added pyridine (27.3. mu.L, 0.338mmol) and acetyl chloride (12. mu.L, 0.169 mmol). The reaction mixture was stirred at room temperature for 16 h. The starting material was still present, so further pyridine (13.65 μ L, 0.169mmol) and acetyl chloride (6 μ L, 0.084mmol) were added and stirring continued for an additional 16 h. The reaction was concentrated in vacuo, then diluted with EtOAc (20mL) and washed with 5% aqueous citric acid (20 mL). The organic extracts were dried over magnesium sulfate and concentrated in vacuo to give a crude residue. Purification by column chromatography using dichloromethane: methanol (100: 0 to 95: 5 to 90: 10) on silica gel gave 43mg (89%) of the title compound as a mixture of epimers as a yellow foam.

¹H NMR(400MHz，CD₃OD)δ1.2-2.2(4H，br，m)，2.2(3H，s)，3.0-4.5(11H，m)，5.1(m，1H)，6.7-7.2(7H，m)，7.7(1H，m)，7.9(1H，m).

LRMS：m/z APCI⁺575[MH⁺]。

Examples 2 to 38

These compounds were prepared by the method of example 1 starting from the appropriate carboxylic acid chlorides and the appropriate precursors listed in the table below.

A is a mixture of epimers; b is a single epimer

Example 39: 6- [ (3R, 4S) -3- (2, 4-difluorophenyl) -4- { [4- (methylsulfonyl) -5-phenyl- 1, 4-diazacyclooctan-1-yl]Carbonyl } pyrrolidin-1-yl]Nicotinonitrile

To a solution of preparation 73 (25mg, 0.05mmol) in DCM (5mL) was added triethylamine (28. mu.L, 0.2mmol) and methanesulfonyl chloride (8. mu.L, 0.1 mmol). The reaction mixture was stirred at room temperature for 72 h. The starting material was still present, so catalytic DMAP (2mg) was added and stirring continued for a further 16 h. The reaction was diluted with DCM (10mL) and washed with water (10 mL). The organic extracts were dried over magnesium sulfate and concentrated in vacuo to give a crude residue. Purification by column chromatography using dichloromethane: methanol (100: 0 to 95: 5) on silica gel gave 16mg (55%) of the title compound as a white solid. The title compound is a single epimer, but the absolute configuration at the point of 5-phenyl substitution is unknown.

¹H NMR(400MHz，CD₃OD)δ1.2-2.2(4H，br，m)，2.34(3H，s)，2.39(3H，s)，3.07(1H，m)，3.4-4.4(12H，m)，6.61(1H，t)，6.93(2H，m)，7.09(1H，d)，7.35(4H，m)，7.5(1H，m)，7.73(1H，m)，8.39(1H，d)。LRMS：APCI⁺m/z 580[MH⁺]。

Examples 40 to 43

These compounds were prepared by the method of example 39 using methanesulfonyl chloride and the appropriate suitable precursors listed in the table below.

A is a mixture of epimers; c-a single epimer with the opposite configuration at the 5-phenyl substitution site.

Example 44: 4- { [ (3S,4R) -1-tert-butyl-4- (2, 4-difluorophenyl) pyrrolidin-3-yl]Carbonyl- 8S- (4-chlorophenyl) -N, N-diethyl-1, 4-diazacyclooctane-1-carboxamide

To a solution of preparation 70 (40mg, 0.082mmol) in pyridine (1mL) was added DMAP (50mg, 0.41mmol) and diethylcarbamoyl chloride (0.103. mu.L, 0.82 mmol). The reaction mixture was stirred in a microwave oven at 120 ℃ for 2h and then cooled to RT over 16 h. A5% citric acid solution (10mL) was added to dilute the reaction and extracted with EtOAc (3X 10 mL). The combined organic extracts were washed with 5% citric acid solution (10mL), sodium bicarbonate solution (10mL) and brine (10mL), dried over sodium sulfate, filtered and concentrated in vacuo to give a crude residue which was purified with AP3(rf 2.68) to give the title compound (15mg ═ 31% yield). LRMS: APCI⁺m/z 589[MH⁺]。

Examples 45 to 46

These compounds were prepared by the method of example 44 using the appropriate commercially available carbamoyl chlorides and the appropriate precursors listed in the table below.

B is a single epimer

Example 47: 8S- (4-chlorophenyl) -4- { [ (3S,4S) -4- (5-chloropyridin-2-yl) -1- (1-methyl-6-) Oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl]Carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid Methyl ester

To a solution of preparation 82 (40mg, 0.074mmol) in DCM (5mL) was added N, N-diisopropylethylamine (51. mu.L, 0.296mmol) and methyl chloroformate (17. mu.L, 0.222 mmol). The reaction mixture was stirred at room temperature for 16 h. Potassium carbonate solution (10mL) was added to dilute the reaction and extracted with DCM (2X 3 mL). The combined organic extracts were concentrated in vacuo to give a crude residue. Purification by column chromatography using ethyl acetate: methanol: 0.88 ammonia (gradient 98: 2: 0.2 to 80: 20: 3) on silica gel gave 37mg (83%) of the title compound as a yellow foam.

¹H NMR(400MHz，CD₃OD)δ1.10-1.30(1H，m)，1.40-1.60(1H，m)，1.70-1.80(1H，m)，1.95-2.05(1H，m)，2.10-2.25(1H，m)，2.70-3.05(2H，m)，3.35-4.10(15H，m)，5.00-5.20(1H，dd)，6.80-6.90(1H，m)，6.95-7.05(2H，m)，7.10-7.40(4H，m)，7.60-7.70(1H，m)，8.00-8.10(1H，s)。LRMS：EI⁺m/z 599[MH⁺]。

Examples 48 to 57

These compounds were prepared by the method of example 47 using the appropriate commercially available chloroformate and the appropriate precursors listed in the following table.

A is a mixture of epimers; b is a single epimer

Examples 58-65 were prepared according to scheme 2.

Example 58: 6- [ (3S,4R) -3- [5S- (4-chlorophenyl) -4- (3,3, 3-trifluoropropionyl) -1, 4-dinitrogen Heterocyclic octane-1-yl]Carbonyl } -4- (2, 4-difluorophenyl) pyrrolidin-1-yl]Pyridazine-3-carbonitriles

To a solution of preparation 81 (15mg, 0.028mmol) in DCM (1mL) was added triethylamine (31. mu.L, 0.224mmol), 3,3, 3-trifluoropropionic acid (9mg, 0.068mmol), and HATU (32mg, 0.084 mmol). The reaction mixture was stirred at room temperature for 16 h. Sodium bicarbonate solution (2mL) was added to dilute the reaction and extracted with DCM (2 mL). The combined organic extracts were concentrated in vacuo to give a crude residue. Purification by AP3 gave 3mg (17%) of the title compound. AP3 Rf ═ 3.5. LRMS: EI (El)⁺m/z 647[MH⁺]。

Example 59: 1- { [ (3S,4R) -1-tert-butyl-4- (2, 4-difluorophenyl) pyrrolidin-3-yl]Carbonyl- 5S- (4-chlorophenyl) -4- (tetrahydro-2H-pyran-4-ylcarbonyl) -1, 4-diazacyclooctane

To a solution of preparation 70 (30mg, 0.061mmol) in DCM (10mL) was added triethylamine (43. mu.L, 0.43mmol), 2-chloro-1, 3-dimethylimidazoline hydrochloride (21mg, 0.122mmol), and tetrahydro-2H-pyran-4-carboxylic acid (40mg, 0.31 mmol). The reaction mixture was stirred at room temperature for 48 h. Sodium bicarbonate solution (2mL) was added to dilute the reaction and the separated organic extracts were concentrated in vacuo to give a crude residue. Purification by AP3 gave 15mg (41% yield) of the title compound (rf 3.43). LRMS: APCI⁺m/z 602[MH⁺]。

Example 60: 1- { [ (3S,4R) -1-tert-butyl-4- (2, 4-difluorophenyl) pyrrolidin-3-yl]Carbonyl- 5S- (4-chlorophenyl) -4- (1-methylcyclopropanecarbonyl) -1, 4-diazacyclooctane

This compound was prepared by the method of example 59 using the appropriate carboxylic acid and the compound of preparation 70. Purification by AP3 gave 16.69mg of the title compound (rf 3.66). LRMS: APCI⁺m/z 572[MH⁺]。

Example 61: 1- { [ (3S,4R) -1-tert-butyl-4- (2, 4-difluorophenyl) pyrrolidin-3-yl]Carbonyl- 5S-4-chlorophenyl) -4- [ (3, 3-difluorocyclobutyl) carbonyl]-1, 4-diazacyclooctane

To a solution of preparation 70 (30mg, 0.061mmol) in DCM (10mL) was added triethylamine (34. mu.L, 0.25mmol), PS-Mukaiyama reagent (144mg, 0.122mmol) and 3, 3-difluorocyclobutanecarboxylic acid (8mg, 0.061 mmol). The reaction mixture was stirred at room temperature for 24 h. The reaction mixture was filtered and the filtrate was concentrated in vacuo. Sodium bicarbonate solution (15mL) was added to dilute the residue and extracted with EtOAc (3X 15 mL). The combined organic extracts were concentrated in vacuo to give a crude residue which was purified by AP3(rf 2.74) to give 1.6mg (4% yield) of the title compound. LRMS: APCI⁺m/z 608[MH⁺]。

Examples 62 to 65

These compounds were prepared by the method of example 61 using the appropriate carboxylic acid and the appropriate precursors listed in the table below.

B is a single epimer

Examples 66-105 were prepared according to scheme 3.

Example 66: (8S-4-chlorophenyl) -4- { [ (3S,4R) -4- (2, 4-difluorophenyl) -1- (1-methyl-6-oxo Sub-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl]Carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester Esters

To a suspension of preparation 60 (200mg, 0.470mmol) in DCM (6mL) was added triethylamine (197. mu.L, 1.41mmol), 1-hydroxybenzotriazole monohydrate (83mg, 0.542mmol) and EDCI (113mg, 0.589 mmol). The reaction mixture was stirred at room temperature for 30 min. The compound of preparation 15a was then added and the reaction mixture was stirred at room temperature for 16 h. The reaction was concentrated in vacuo, and the residue was partitioned between EtOAc (20mL) and citric acid solution (10 mL). The organic layer was separated, washed with sodium bicarbonate solution (10mL), dried over magnesium sulfate, and concentrated in vacuo to give a crude residue. Purification by column chromatography using dichloromethane: methanol: 0.88 ammonia (gradient from pure dichloromethane to 95: 5: 0.5) on silica gel gave 234mg (83%) of the title compound as a yellow foam.

¹H NMR(400MHz，CD₃OD)δ0.95-1.10(1H，m)，1.45-1.60(1H，m)，1.65-1.75(1H，m)，1.90-2.05(1H，m)，2.10-2.25(1H，m)，2.35-2.50(1H，m)，2.85-3.05(1H，m)，3.30-4.10(15H，m)，4.95-5.15(1H，dd)，7.10-7.35(4H，m)，6.95-7.05(4H，m)，7.40-7.50(1H，m)。LRMS：EI⁺m/z 600[MH⁺]。

Examples 67 to 97

These compounds were prepared by the method of example 66 using the appropriate carboxylic acid and the appropriate precursors listed in the table below.

A is a mixture of epimers; b is a single epimer

Example 98: 8S- (4-chlorophenyl) -4- { [ (3S,4S) -4- (5-chloropyridin-2-yl) -1- (6-cyanopyridazine -3-yl) pyrrolidin-3-yl]Carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester

To a solution of preparation 65 (24mg, 0.075mmol) in DCM (3mL) was added diisopropylethylamine (79 μ L, 0.46mmol), HBTU (47mg, 0.13mmol) and preparation 15a (40mg, 0.13 mmol). The reaction mixture was stirred at room temperature for 24h, then diluted by addition of sodium bicarbonate solution (2 mL). The separated organic extract was concentrated in vacuo to give a crude residue which was purified with AP3(rf 3.67) to give 42mg (64% yield) of the title compound. LRMS: APCI⁺m/z 594[MH⁺]。

Examples 99 to 105

These compounds were prepared by the method of example 98, starting from the appropriate precursors listed in the table below.

A is a mixture of epimers; b is a single epimer

Example 104: 1- { [ (3S,4R) -1-tert-butyl-4- (2, 4-difluorophenyl) pyrrolidin-3-yl]Carbonyls -4-butynyl-5-phenyl-1, 4-diazacyclooctane

To a solution of preparation 100 (24mg, 0.075mmol) in dimethylacetamide (1.25mL) was added triethylamine (42 μ L, 0.30mmol), TBTU (24mg, 0.075mmol) and preparation 21 (12mg, 0.050 mmol). The reaction mixture was shaken at 60 ℃ for 24 h. The reaction mixture was concentrated in vacuo to give a crude residue. Purification by AP3(rf 3.68) gave 7mg (27% yield) of the title compound as a mixture of epimers. LRMS: APCI⁺m/z 526[MH⁺]。

Example 105-108 was prepared according to scheme 4.

Example 105: 6- [ (3S,4R) -3- { [5- (4-chlorophenyl) -4- (methylsulfonyl) -1, 4-diazacyclo Octane-1-yl]Carbonyl } -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-1-yl]Pyridazine-3-carbonitriles

To a solution of preparation 92 (30mg, 0.057mmol) in acetonitrile (10mL) was added 3-chloro-6-cyanopyridazine (12mg, 0.086mmol) and N, N-diisopropylethylamine (40. mu.L, 0.23 mmol). The reaction mixture was stirred at reflux for 3 h. The reaction was concentrated in vacuo and sodium bicarbonate solution (10mL) was added to dilute the residue and extracted with EtOAc (3X 10 mL). The combined organic extracts were wee washed with brine (15mL), dried with sodium sulfate, filtered and concentrated in vacuo to give a crude residue. Purification by column chromatography on silica gel using dichloromethane: methanol: 0.88 ammonia (95: 5: 0.5) gives 25mg (78%) of the title compound as a white solid as a mixture of epimers.

¹H NMR(400MHz，CD₃OD)δ1.19-2.47(10H，m)，2.87-4.49(12H，m)，4.80-4.85(1H，m)，6.52-6.69(3H，m)，6.92-6.94(2H，m)，7.09-7.37(3H，m)，7.42-7.47(1H，m)。LRMS：APCI⁺m/z 627[MH⁺]。

Example 106: 8- (4-chlorophenyl) -4- { [ (3S,4S) -4- (2-fluoro-4-methoxyphenyl) -1- (6-cyanopyridaz-d Oxazin-3-yl) pyrrolidin-3-yl]Carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester

The compound was prepared according to the method of example 105, but starting from 3-chloro-6-cyanopyridazine and the compound of preparation 93. LRMS: APCI⁺m/z 607[MH⁺]。

Example 107: 8- (4-chlorophenyl) -4- { [ (3S,4R) -1- (6-chloropyridazin-3-yl) -4- (2-fluoro-4-methyl-l Oxyphenyl) pyrrolidin-3-yl]Carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester

To a solution of the compound of preparation 93 (65mg, 0.13mmol) in dimethyl sulfoxide (2mL) were added 3, 6-dichloropyridazine (58mg, 0.39mmol), cesium fluoride (20mg, 0.13mmol) and triethylamine (54. mu.L, 0.39 mmol). The reaction mixture was stirred at reflux for 24h and then cooled to RT over 40 h. The reaction was diluted with sodium bicarbonate solution (30mL) and extracted with diethyl ether (4X 20 mL). The combined organic extracts were washed with brine (3 × 25mL) and concentrated in vacuo to give a crude residue. Purification by column chromatography using dichloromethane: methanol: 0.88 ammonia (98: 2: 0.2) on silica gel gave 73mg (92%) of the title compound as a mixture of epimers as an off-white foam.

¹H NMR(400MHz，CD₃OD)δ0.92-2.33(5H，m)，2.56-3.76(14H，m)，3.85-4.36(3H，m)，4.85-5.20(1H，m)，6.40-6.64(3H，m)，6.77-6.89(1H，m)，6.99-7.25(5H，m)。LRMS：APCI⁺m/z 616[MH⁺]。

Example 108: [5- (4-chlorophenyl) -4- (methylsulfonyl) -1, 4-diazacyclooctan-1-yl]- [ (3S,4R) -1- (6-chloro-pyridazin-3-yl) -4- (2-fluoro-4-methoxyphenyl) -pyrrolidin-3-yl]-methanones

The compound was prepared by the method of example 107 but starting from 3, 6-dichloropyridazine and the compound of preparation 92. LRMS: APCI⁺m/z 636[MH⁺]。

Example 109-110

These compounds were prepared by the method of example 107 using 3, 6-dichloropyridazine and the appropriate precursors listed in the table below.

A single epimer with an unknown absolute configuration at the C ═ chloro-phenyl substitution site; single epimer with the opposite configuration at the chloro-phenyl substitution site relative to the compound of example 109

Example 111-114 was prepared according to scheme 5.

Example 111: 6- [ (3S,4R) -3- { [5- (4-chlorophenyl) -4- (methylsulfonyl) -1, 4-diazacyclo Octane-1-yl]Carbonyl } -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-1-yl]Pyridazin-3 (2H) -ones

The compound (100mg, 0.157mmol) from example 108 was dissolved in acetic acid (5 mL). The resulting solution was degassed thoroughly and stirred under nitrogen at reflux overnight. The reaction was concentrated in vacuo, and the residue was diluted with sodium bicarbonate solution (25mL) and extracted with EtOAc (3X 25 mL). The combined organic extracts were washed with sodium bicarbonate solution (30mL), brine (30mL), dried over sodium sulfate, filtered and concentrated in vacuo to give a crude residue. Purification by column chromatography using dichloromethane: methanol: 0.88 ammonia (92.5: 7.5: 0.75) on silica gel gave 90mg (93%) of the title compound as a mixture of epimers as an off-white foam.

¹H NMR(400MHz，CD₃OD)δ1.09-2.53(8H，m)，2.91-3.23(1H，m)，3.34-4.38(14H，m)，6.60-6.77(2H，m)，6.83-7.09(2H，m)，7.26-7.40(5H，m)。LRMS：APCI⁺m/z 618[MH⁺]。

Example 112-

These compounds were prepared by the method of example 111, starting from the appropriate precursors listed in the table below.

A is a mixture of epimers; d ═ a single epimer of unknown absolute configuration at the chlorophenyl substitution site; single epimer with the opposite configuration at the chlorophenyl substitution site relative to the compound of example 113

Example 115-119 was prepared according to scheme 6.

Example 115: 6- [ (3S,4R) -3- { [5- (4-chlorophenyl) -4- (methylsulfonyl) -1, 4-diazacyclo Octane-1-yl]Carbonyl } -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-1-yl]-2-methylpyridazin-3 (2H) - Ketones

To a solution of the compound from example 111 (45mg, 0.073mmol) in DMF (2mL) was added lithium bromide (7.6mg, 0.087mmol) and sodium hexamethyisilazide (16mg, 0.087 mmol). the reaction mixture was stirred at room temperature for 30 min. iodomethane (5.4. mu.L, 0.087mmol) was added, the resulting solution was stirred at RT for 24 h. the reaction was concentrated in vacuo, sodium bicarbonate solution (20mL) was added to dilute the residue, washed with ethyl acetate (4X 20mL), the combined organic extracts were washed with brine (20mL) and concentrated in vacuo to give a crude residue, which was purified by column chromatography using dichloromethane: methanol: 0.88 ammonia (95: 5: 0.5) on silica gel to give 46mg (59%) of the title compound as a mixture of epimers which was a foam.

¹H NMR(400MHz，CD₃OD)δ1.09-1.34(2H，m)，1.43-2.53(6H，m)，2.85-3.23(1H，m)，3.34-4.38(17H，m)，6.60-6.76(2H，m)，6.87-6.91(1H，m)，7.01-7.09(2H，m)，7.24-7.40(5H，m)。LRMS：APCI⁺m/z 632[MH⁺]。

Example 116-

These compounds are prepared by the method of example 121 starting from the appropriate precursors listed in the table below.

A is a mixture of epimers; b is a single epimer; d-a single epimer of unknown absolute configuration at the chlorophenyl substitution site

Preparation of

Preparation 1: 3-chloro-1- (2, 4-difluorophenyl) propan-1-one

To a stirred mixture of aluminum (III) chloride (11.70g, 88.0mmol) in 1, 3-difluorobenzene (21mL) was added 3-chloropropionyl chloride (4.0mL, 41.9mmol) at RT. The reaction mixture was stirred at 60 ℃ for 6 h. The mixture was cooled to RT and poured into an ice/2M aqueous hydrochloric acid mixture (100 mL). After vigorous stirring, the mixture was extracted with DCM (2X 75 ml). The combined organic extracts were dried over magnesium sulfate and concentrated in vacuo to give 8.68g (quantitative yield) of the crude title compound as an orange oil.

¹H NMR(400MHz，CDCl₃)δ3.35-3.45(2H，t)，3.80-3.90(2H，t)，6.80-6.90(1H，m)，6.90-7.00(1H，m)，7.90-8.00(1H，m).

Preparation 2: (±) -1-benzyl-5- (2, 4-difluorophenyl) -1, 4-diazepane

A solution of N-benzylethylenediamine (2.50g, 16.64mmol) in 4-methyl-pentan-2-one (50mL) was stirred at reflux under Dean-Stark conditions for 3 h. The solution was cooled to RT and triethylamine (3.48mL, 25.0mmol) and the compound of preparation 1 (3.75g, 18.3mmol) were added thereto. The reaction mixture was stirred at 60 ℃ for 16 h. The mixture was cooled to RT and concentrated in vacuo. The residue was dissolved in a mixture of propan-2-ol and ater (50mL, 95: 5) and stirred at RT for 16 h. Sodium borohydride (1.50g, 39.65mmol) was added and the reaction mixture was stirred at RT for 16 h. Water (100mL) was added to dilute the reaction and propan-2-ol was removed in vacuo. The aqueous mixture was extracted with DCM (3X 50 mL). The combined organic extracts were dried over magnesium sulfate and concentrated in vacuo to give a crude residue. Purification by column chromatography using ethyl acetate: methanol: 0.88 ammonia (gradient 98: 2: 0.2 to 80: 20: 3) on silica gel gave 2.28g (45%) of the racemic title compound as a yellow oil.

¹H NMR(400MHz，CDCl₃)δ1.85-1.95(1H，m)，2.00-2.10(1H，m)，2.60-2.75(2H，m)，2.75-2.90(2H，m)，2.95-3.05(1H，m)，3.10-3.20(1H，m)，3.68(2H，s)，4.25-4.35(1H，m)，6.70-6.80(1H，m)，6.80-6.90(1H，m)，7.20-7.40(5H，m)，7.40-7.50(1H，m)。LRMS：APCI⁺m/z 303[MH⁺]。

Preparation 3: (±) -1-benzyl-5- (4-chlorophenyl) -1, 4-diazepane

This compound was prepared according to preparation 2 starting from N-benzylethylenediamine, except that commercially available 3-chloro-1- (4-chlorophenyl) propan-1-one was used. LRMS: APCI⁺m/z 300[MH⁺]。

Preparation 4: (±) -5- (2, 4-difluorophenyl) -1, 4-diazepane

To a solution of preparation 2 compound (220mg, 0.728mmol) in ethanol (20mL) was added 20% palladium (II) hydroxide on carbon catalyst (102mg, 0.146mmol) and ammonium formate (229mg, 3.64 mmol). The reaction mixture was stirred at 75 ℃ for 3 h. The solution was cooled to RT and used under nitrogenThe catalyst was filtered off. The catalyst was washed with a further 15ml of ethanol and the combined filtrates were concentrated in vacuo to give 172mg (quantitative yield) of the crude title compound (racemate) as a colourless oil.

¹H NMR(400MHz，CD₃OD)δ1.85-1.95(1H，m)，2.00-2.10(1H，m)，2.85-3.05(4H，m)，3.05-3.15(2H，m)，4.10-4.20(1H，m)，6.85-6.95(2H，m)，7.40-7.50(1H，m)。LRMS：APCI⁺m/z 213[MH⁺]。

Preparation 5: (±) {7a- (4-chlorophenyl) hexahydro-5H-pyrrolo [1, 2-a)]Imidazole-5-one }

To a solution of 3- (4-chlorobenzoyl) propionic acid (40g, 190mmol) in xylene (250mL) was added ethylenediamine (22.6g, 376mol) and 12M hydrochloric acid (0.500mL, 6 mmol). The reaction mixture was stirred at reflux for 12h under Dean-Stark and then cooled to room temperature. The solid was collected by filtration and washed with xylene to give a crude product. It was dissolved in DCM (400ml) and filtered. The filtrate was concentrated in vacuo to give 36.6g (62%) of the racemic title compound as a beige solid.

¹H NMR(400MHz，CD₃Cl)δ2.22-2.35(2H，m)，2.49-2.56(1H，m)，2.78-2.95(3H，m)，3.27-3.34(1H，m)，3.68-3.77(1H，m)，7.32-7.35(2H，m)，7.42-7.46(7.42-7.46(2H，m)。LRMS：APCI⁺m/z 237[MH⁺]。

Preparation 6: (±) [5- (4-chlorophenyl) -1, 4-diazacyclooctane]

Lithium aluminum hydride (3.21g, 84.5mmol) was added to a flask containing the compound of preparation 5 (5.00g, 21.1mmol), and diethyl ether (120ml) was added. The reaction mixture was stirred at room temperature for 30min, then refluxed for 72 h. The reaction was cooled in an ice bath and quenched by the addition of water (3.2ml), 2M sodium hydroxide solution (3.2ml) and water (9.6 ml). Reaction mixture is passed throughFiltered and washed with diethyl ether (25 ml). The filtrate was concentrated in vacuo to give a crude residue. Purification by column chromatography on silica gel using dichloromethane: methanol: 0.88 ammonia (75: 25: 2.5) gave 2.3g (45%) of the racemic title compound as a colorless oil.

¹H NMR(400MHz，CD₃Cl)δ1.56-1.83(3H，m)，1.96-2.04(1H，m)，2.74-3.10(6H，m)，3.84-3.88(1H，m)，7.26-7.31(4H，m)。LRMS：APCI⁺m/z 225[MH⁺]。

Preparation 7 to 11

These compounds were prepared by the methods of preparation 5 and 6 from ethylenediamine and commercially available propionic acid in analogy to the user in preparation 5. Each compound was obtained as a racemate.

Preparation 12: (S) and (R) -5- (4-chlorophenyl) -1, 4-diazacyclooctane

Chiral resolution of 12g of the racemic compound of preparation 6 was initially carried out by column chromatography on a 500 x 50mm internal diameter Chiralcel OD-H column eluting with heptane: isopropanol: diethylamine (80: 20: 0.1). This gave 5.74g (96% of theory) of the first eluting enantiomer (preparation 12a), which was a colorless oil at 99.4% ee, and 5.13g (86% of theory) of the second eluting enantiomer (preparation 12b), which was a colorless oil at 95.5% ee. For analytical purposes, the Chiralcel OD-H250X 4.6 column eluted the enantiomer at 1ml/min with retention times of 6.10 and 8.68 min, respectively. Each shows the same proton NMR spectrum:¹H NMR(400MHz，CD₃Cl)δ1.56-1.83(3H，m)，1.96-2.04(1H，m)，2.74-3.10(6H，m)，3.84-3.88(1H，m)，7.26-7.31(4H，m)。LRMS：APCI⁺m/z 225[MH⁺]。

or alternatively, the salt formation of the racemic compound of preparation 6 with a chiral acid, which salt formation allows the fractional crystallization of the individual diastereomeric salts 12c, with the advantage that the absolute configuration is subsequently allowed to be indicated.

Thus, 5.0g of the racemic compound of preparation 6 was dissolved in tert-butyl methyl ether (150mL), the solution was heated to 57 ℃ and 75mL of the solvent was evaporated off under atmospheric pressure. 45ml of ethanol were added and 65ml of the solution were evaporated off again. When cooled to 45 ℃ 8.2g of di-p-benzoyl-L-tartaric acid were charged in one portion, the suspension was heated again to 45 ℃ and the solution was cooled to 20 ℃ over 3 hours and then granulated for a further 13 hours. Filtration afforded the di-p-benzoyltartrate salt 12c (8.5g) as a white solid which was recrystallized from 10% v/v water/methanol on heating to give a material of > 98% ee. The absolute configuration shown by X-ray crystallography is the diazacyclooctane-S-enantiomer. Chiral HPLC analysis of 12c as "free base" showed it to correspond to enantiomer 12 a.

Preparation 13: (±) -tert-butyl 5- (4-chlorophenyl) -1, 4-diazacyclooctane-1-formate

Triethylamine (0.285mL, 2.05mmol) was added to a solution of preparation 6 (460mg, 2.05mmol) in THF (10mL), di-tert-butyl dicarbonate (536mg, 2.46mmol) was added, and the resulting solution was stirred at RT for 16 h. The reaction was concentrated in vacuo, and sodium bicarbonate solution (10mL) was added to dilute the residue, which was washed with EtOAc (4X 20 mL). The combined organic extracts were concentrated in vacuo to give a crude residue. Purification by column chromatography using EtOAc on silica gel gave 419mg (63%) of the racemic title compound as a colourless oil.

¹H NMR(400MHz，CDCl₃)δ1.47-1.49(9H，m)，1.54-1.70(2H，m)，1.74-1.81(1H，m)，1.87-1.98(1H，m)，2.85-2.94(1H，m)，2.97-3.12(2H，m)，3.29-3.51(1H，m)，3.59-3.69(1H，m)，3.72-3.80(1H，m)，3.82-3.96(1H，m)，7.23-7.28(4H，m)。LRMS：EI⁺m/z 325[MH⁺]。

Enantiomers 13a and 13b were similarly prepared starting from the compounds of preparation 12a and 12b, respectively.

Preparation 14: (±) -1-tert-butyl 4-methyl 5- (4-chlorphenyl) -1, 4-diazacyclooctane-1, 4-dimethyl Acid esters

To a stirred solution of preparation 13 compound (469mg, 1.44mmol) in pyridine (15mL) was added triethylamine (805. mu.L, 5.77mmol), methyl chloroformate (781. mu.L, 10.12mmol) and DMAP (352mg, 2.88 mmol). The reaction was stirred at 60 ℃ for 16 h. The reaction was cooled to RT and quenched by addition of potassium carbonate solution (10 mL). The reaction mixture was concentrated in vacuo and the residue was diluted with (15 mL). The aqueous compound was extracted with DCM (3X 10 mL). The combined organic extracts were concentrated in vacuo to give a crude residue. Purification by column chromatography using pentane: ethyl acetate (gradient 3: 1 to pure EtOAc) on silica gel afforded 322mg (58%) of the racemic title compound as a colorless oil.

¹H NMR(400MHz，CDCl₃)δ1.35-1.50(9H，s)，1.55-1.65(1H，m)，1.80-1.95(2H，m)，2.05-2.30(1H，m)，2.85-3.15(3H，m)，3.40-3.50(1H，m)，3.55-3.65(1H，m)，3.65-3.80(3H，m)，3.90-4.00(1H，m)，5.00-5.35(1H，m)，7.05-7.30(4H，m)。LRMS：APCI⁺m/z 283[M-Boc+H]⁺.

14a and 14b were prepared similarly starting from the compounds of preparation 13a and 13b, respectively.

Preparation 15: (±) -methyl 8- (4-chlorophenyl) -1, 4-diazacyclooctane-1-formate

To a stirred solution of preparation 14 (322mg, 0.841mmol) in DCM (5mL) was added 4M HCl in 1, 4-dioxane (5 mL). The reaction was stirred at RT for 16 h. The reaction mixture was concentrated in vacuo and the residue was washed with potassium carbonate solution (15 mL). The aqueous compound was extracted with DCM (3X 10 mL). The combined organic extracts were concentrated in vacuo to give 230mg (97%) of the racemic title compound as a yellow oil.

¹H NMR(400MHz，CDCl₃)δ1.75-2.05(2H，m)，2.25-2.40(1H，m)，2.65-2.85(2H，m)，2.95-3.05(2H，m)，3.05-3.25(2H，m)，3.40-3.50(1H，m)，3.65-3.75(3H，m)，5.05-5.30(1H，dd)，7.10-7.30(4H，m)。LRMS：APCI⁺m/z 283[MH⁺]。

Enantiomers 15a and 15b were prepared starting from the compounds of preparation 14a and 14b, respectively.

Preparation 16: (±) -5- (4-chlorophenyl) -4- (methylsulfonyl) -1, 4-diazacyclooctane-1-carboxylic acid tert-butyl ester Butyl ester

DMAP (2.67g, 21.9mmol) was added to a solution of preparation 13 compound (2.37g, 7.30mmol) in pyridine (20mL), methanesulfonyl chloride (1.69mL, 21.9mmol) was added and the resulting solution was stirred at 50 ℃ for 3 h. The reaction was concentrated in vacuo and 10% citric acid solution (50mL) was added to dilute the residue and extracted with EtOAc (4X 50 mL). The combined organic extracts were washed with 10% citric acid (50ml), brine (50ml), dried over sodium sulphate, filtered and concentrated in vacuo to give a crude residue. Purification by column chromatography on silica gel using pentane: ethyl acetate (1: 1) gave 1.69g (58%) of the racemic title compound as a colourless oil.

¹H NMR(400MHz，CDCl₃)δ1.47-1.49(9H，m)，1.70-1.77(2H，m)，1.98-2.06(1H，m)，2.12-2.30(1H，m)，2.40-2.43(3H，m)，3.11-3.30(1H，m)，3.34-3.46(2H，m)，3.52-3.60(1H，m)，3.73-3.89(1H，m)，4.07-4.15(1H，m)，4.91-4.98(1H，m)，7.22-7.26(2H，m)，7.31-7.35(2H，m)。LRMS：APCI⁺m/z 403[MH⁺]。

Preparation 17: (±) -8- (4-chlorophenyl) -1- (methylsulfonyl) -1, 4-diazacyclooctane hydrochloride

4M HCl in dioxane (20mL, 80mmol) was added to the compound of preparation 16 (1.69g, 4.19mmol) and the resulting solution was stirred at RT for 16 h. The reaction was concentrated in vacuo to give 1.42g (100%) of the racemic title compound as an off-white solid.

¹H NMR(400MHz，CDCl₃)δ1.92-2.12(2H，m)，2.22-2.43(4H，m)，2.86-3.01(1H，m)，3.36-3.80(5H，m)，4.14-4.26(1H，m)，4.49-5.07(1H，m)，7.30-7.39(4H，m)。LRMS：APCI⁺m/z 303[MH⁺]。

Preparation 17a-26

These compounds were prepared by the methods of preparation 13-17, starting from the appropriate precursors listed in the table below.

(1) A single enantiomer; (2) racemic modification

Preparation 27: (4S) -4-benzyl-3- [ (2E) -3- (2-fluoro-4-methoxyphenyl) prop-2-enoyl]-1, 3-oxa Oxazolidin-2-ones

To a commercially available solution of 2-fluoro-4-methoxyphenyl cinnamic acid (16.5g, 84.1mmol) in DCM (100mL) at 4 deg.C was added DMF (0.1mL) followed by a dropwise addition of oxalyl chloride (14.8mL, 170mmol) in DCM (50 mL). The reaction mixture was warmed to RT over 3 h. The reaction was concentrated in vacuo and azeotroped with DCM (2 × 100mL) to give the crude intermediate acid chloride. The acid chloride was dissolved in DCM (50mL) and added dropwise to an ice-cooled solution of(s) - (-) -4-benzyl-2-oxazolidinone (14.3g, 80.7mmol), lithium chloride (17.8g, 421mmol) and triethylamine (58.8mL, 421mmol) in DCM (100 mL). The reaction was stirred at RT for 16h, then diluted with water (100mL) and passedAnd (5) filtering. The filtrate was partitioned and the aqueous phase extracted with DCM (2X 100 mL). The combined organic extracts were dried over sodium sulfate, filtered and concentrated in vacuo to give a crude residue. Subjecting the residue to reaction in a second stepTriturated in ether (150 mL). Filtration gave 20g (67%) of the title compound as a light brown solid.

¹H NMR(400MHz，CDCl₃)δ2.82-2.88(1H，m)，3.36-3.40(1H，m)，3.84(3H，s)，4.18-4.27(2H，m)，4.77-4.83(1H，m)，6.63-6.67(1H，m)，6.72-6.76(1H，m)，7.23-7.37(5H，m)，7.60-7.64(1H，m)，7.84-7.88(1H，m)，8.01-8.05(1H，m)。LRMS：APCI⁺m/z 356[MH⁺]。

Preparation 28: (4S) -4-benzyl-3- [ (2E) -3- (4-cyanophenyl) prop-2-enoyl]-1, 3-oxazolidine- 2-ketones

The compound was classified by the method of preparation 27, but starting with commercially available 4-cyanocinnamic acid. LRMS: APCI⁺m/z 333[MH⁺]。

Preparation 29: (4S) -4-benzyl-3- { [ (3S,4R) -1-benzyl-4- (2-fluoro-4-methoxyphenyl) pyrrolidine- 3-yl]Carbonyl } -1, 3-oxazolidin-2-ones

To a solution of preparation 27 (20g, 56.3mmol) in DCM (200mL) at 5 deg.C was added trifluoroacetic acid (0.347mL, 67.5mmol) followed by the dropwise addition of commercially available N- (methoxymethyl) -N- (trimethylsilylmethyl) benzylamine (16g, 67.5 mmol). The reaction mixture was stirred at room temperature for 64h, then diluted by addition of sodium bicarbonate solution (100 mL). The organic extract was concentrated in vacuo to give a crude residue. Purification by column chromatography using ethyl acetate: dichloromethane (1: 1) on silica gel gave 12g (43%) of the title compound as an oil and as the single diastereomer shown.

¹H NMR(400MHz，CDCl₃)δ2.70-2.89(3H，m)，3.12-3.28(3H，m)，3.59-3.80(5H，m)，4.09-4.26(3H，m)，4.29-4.39(1H，m)，4.64-4.72(1H，m)，6.56-6.62(1H，m)，6.65-6.70(1H，m)，7.10-7.15(2H，m)，7.21-7.40(9H，m)。LRMS：APCI⁺m/z 489[MH⁺]。

Preparation of 30 to 32

These compounds were prepared by the method of preparation 29 using the appropriate precursors listed in the table below. Each compound was obtained as a single diastereomer.

Preparation 33: (3S,4R) -1-benzyl-4- (2-fluoro-4-methoxyphenyl) pyrrolidine-3-carboxylic acid methyl ester

To a stirred solution of the compound of preparation 29 (12.00g, 24.56mmol) in dry methanol (100mL) was added samarium (III) triflate (1.17g, 1.96mmol) portionwise at RT. The reaction was stirred at RT under nitrogen for 16 h. The reaction mixture was concentrated in vacuo to give a crude residue. Purification by column chromatography using pentane: ethyl acetate (gradient 4: 1 to 1: 1) on silica gel afforded 6.50g (77%) of the title compound as a single diastereomer as a pale yellow oil.

¹H NMR(400MHz，d⁶-DMSO)δ2.55-2.65(1H，m)，2.90-3.05(3H，m)，3.10-3.20(1H，m)，3.60-3.65(4H，m)，3.70-3.85(5H，m)，6.60-6.65(1H，d)，6.70-6.75(1H，d)，7.20-7.40(6H，m)。LRMS：APCI⁺m/z 344[MH]⁺.

Preparation of 34 to 36

These compounds were prepared by the method of preparation 33, starting with the precursors listed in the table below. Each compound is obtained as a single enantiomer.

Preparation 37: (3S,4R) -4- (2-fluoro-4-methoxyphenyl) pyrrolidine-3-carboxylic acid methyl ester

To a solution of preparation 33 (1.8g, 5.2mmol) in methanol (20mL) was added 20% palladium hydroxide on carbon (180mg) and 1-methyl-1, 4-cyclopentadiene (2.94mL, 26.2 mmol). The reaction mixture was stirred at reflux for 2.5h and then cooled to RT. Reactant is throughFiltered and washed with methanol. The filtrate was concentrated in vacuo to give 1.30g (98% yield) of the title compound as a colorless oil.

¹H NMR(400MHz，CD₃OD)δ2.83-2.89(1H，m)，3.09-3.15(1H，m)，3.20-3.25(1H，m)，3.29-3.37(2H，m)，3.59-3.66(1H，m)，3.64(3H，s)，3.77(3H，s)，6.64-6.72(2H，m)，7.20-7.24(1H，m)。LRMS：APCI⁺m/z 254[MH⁺]。

Preparation 38: (3S,4S) -4- (5-Chloropyridin-2-yl) -1- [1,2,4]Triazolo [4,3-b]Pyridazin-6-yl pyrindine Pyrrolidine-3-carboxylic acid methyl ester

To a solution of preparation 104 (970mg, 3.50mmol) in n-butanol (20mL) was added diisopropylethylamine (2.44mL, 14.0mmol) followed by 6-chloro [1,2,4] triazolo [4,3-b ] pyridazine (811mg, 5.25 mmol). The resulting solution was warmed to 120 ℃ and held for 2 h. The reaction mixture was concentrated in vacuo. The residue was partitioned between brine (30mL) and EtOAc (50 mL). The organic extracts were dried over magnesium sulfate, filtered and concentrated in vacuo to give a crude residue. Purification by column chromatography on silica gel using dichloromethane: methanol (95: 5) gave 1.53g (100%) of the title compound as a single enantiomer as an oily solid.

¹H NMR(400MHz，CH₃OD)δ3.65(3H，s)，3.67-3.84(3H，m)，3.93-4.00(1H，m)，4.03-4.10(2H，m)，7.07-7.11(1H，m)，7.37-7.42(1H，m)，7.75-7.80(1H，m)，7.86-7.91(1H，m)，8.49-8.52(1H，m)，8.95(1H，s)。LRMS：APCI⁺m/z 359[MH⁺]。

Preparation 39: (3S,4S) -4- (2, 4-difluorophenyl) -1- [1,2,4]Triazolo [4,3-b]Pyridazin-6-yl pyrindine Pyrrolidine-3-carboxylic acid methyl ester

This compound was prepared by the method of preparation 38, starting from the same chlorinated heterocycle and the compound of preparation 105. LRMS: APCI⁺m/z 360[MH⁺]。

Preparation 40 to 43

These compounds were prepared by the method of example 105 starting from commercially available 3-chloro-6-cyanopyridazine and the appropriate precursors listed in the table below. Each compound is obtained as a single enantiomer.

Preparation of 44 to 45

These compounds were prepared by the method of example 107, starting from commercially available 3, 6-dichloropyridazine and the appropriate precursors listed in the table below. Each compound is obtained as a single enantiomer.

Preparation of 46-47

These compounds were prepared by the method of example 111 by refluxing the compounds of preparations 44 and 45, respectively, in acetic acid. Each compound is obtained as a single enantiomer.

Preparation 48: (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazine- 3-Yl) pyrrolidine-3-carboxylic acid methyl ester

This compound was prepared by the method of example 115 using the compound of preparation 46, which is a single enantiomer. LRMS: APCI⁺m/z 362[MH⁺]。

Preparation 49: (3S,4R) -4- (2, 4-difluorophenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3- Yl) pyrrolidine-3-carboxylic acid methyl ester

To a stirred mixture of preparation 105 compound (25.27g, 82mmol) and 6-chloro-2-methylpyridazin-3 (2H) -one { Helvetica Chimica Acta; (1954),37837-48} (12.0g, 83.0mmol) to a suspension in degassed toluene (500mL) was added cesium carbonate (111g, 339mmol) and (9, 9-dimethyl-9H-xanthene-4, 5-diyl) -bis [ diphenylphosphine](6.23g, 10.8 mmol). The reaction mixture was purged twice with nitrogen. Palladium (II) diacetate (813mg, 3.62mmol) was added and the reaction stirred under nitrogen at 115 ℃ for 16 h. The reaction mixture was filtered under reduced pressure and the residue was washed with 20ml of toluene. The filtrate was concentrated in vacuo to give a crude product residue. Purification by column chromatography on silica gel using heptane: ethyl acetate (gradient 7: 3 to pure EtOAc to EtOAc: MeOH, 95: 5) gave 23.85g (83%) of the title compound as a yellow oil.

¹H NMR(400MHz，CDCl₃)δ3.30-3.40(1H，m)，3.40-3.50(1H，m)，3.62(3H，s)，3.67(3H，s)，3.62-3.70(1H，m)，3.75-3.90(2H，m)，3.90-4.00(1H，m)，6.75-6.95(4H，m)，7.15-7.25(1H，m)。LRMS：APCI⁺m/z 350[MH]⁺.

Preparation 50: (3S,4R) -4- (5-Chloropyridin-2-yl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3- Yl) pyrrolidine-3-carboxylic acid methyl ester

This compound is prepared by the method of preparation 49 using the same chlorinated heterocycle and the compound of preparation 104, which is a single enantiomer. LRMS: APCI⁺m/z 348[MH]⁺.

Preparation 51: (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1-pyridazin-3-ylpyrrolidine-3-carboxylic acid methyl ester

To a stirred suspension of preparation 44 (407mg, 1.11mmol) in methanol (5mL) was added 20% palladium (II) hydroxide over carbon catalyst (27mg, 0.189mmol) and 1-methyl-1, 4-cyclohexadiene (438 μ L, 3.89 mmol). The reaction mixture was stirred at reflux under nitrogen for 3h and then allowed to cool to RT over 16 h. Under nitrogen atmosphereThe catalyst was filtered off. The catalyst was washed with an additional 10ml of methanol and the combined filtrates were concentrated in vacuo to give 438mg (quantitative yield) of the crude product as a colourless oil which was used directly in the ester hydrolysis for preparation 63.

¹H NMR(400MHz，CDCl₃)δ3.45-3.55(3H，m)，3.68(3H，s)，3.77(3H，s)，3.90-4.05(2H，m)，4.05-4.25(1H，m)，6.60-6.70(2H，m)，7.05-7.15(1H，t)，7.20-7.25(1H，m)，7.75-7.80(1H，m)，8.60-8.65(1H，d)。LRMS：EI⁺m/z 332[MH⁺]。

Preparation 52: (3S,4R) -1- (6-Cyanopyridazin-3-Yl) -4- (2, 4-Difluorophenyl) pyrrolidine-3-carboxylic acid

To a stirred solution of preparation 41 (3.82g, 11.1mmol) in 1, 4-dioxane (100mL) and water (50mL) was added dropwise at 5 deg.C a 1M aqueous solution of sodium hydroxide (9.98mL, 9.98 mmol). The reaction was stirred at room temperature for 16h, then neutralized with 2M aqueous hydrochloric acid (4.99mL, 9.98mmol) and concentrated in vacuo. The crude product residue was azeotroped with toluene (3 × 50mL) to give 3.97g (92%) of the title compound as a cream solid and containing 1 equivalent of sodium chloride by-product.

¹H NMR(400MHz，d⁶-DMSO)δ3.40-3.50(2H，m)，3.65-3.80(1H，m)，3.85-4.20(3H，m)，7.00-7.10(2H，m)，7.15-7.25(1H，m)，7.45-7.55(1H，m)，7.80-7.85(1H，d)。LRMS：APCI⁺m/z 331[MH]⁺.

Preparation 53-65

These compounds were prepared by the method of preparation 52 starting from the appropriate precursors listed in the table below. Each compound is obtained as a single enantiomer.

Preparation 66: 6- [ (3S,4R) -3- { [5S- (4-chlorophenyl) -1, 4-diazacyclooctan-1-yl]Carbonyl- 4- (2, 4-difluorophenyl) pyrrolidin-1-yl]-2-methylpyridazin-3 (2H) -one

To a solution of preparation 60 (200mg, 0.473mmol) in DCM (15mL) was added N, N-diisopropylethylamine (328. mu.L, 1.89mmol), 1-hydroxybenzotriazole monohydrate (83mg, 0.544mmol), and 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide methiodide (176mg, 0.591 mmol). The reaction mixture was stirred at room temperature for 30 min. The compound of preparation 12a was then added and the reaction mixture was stirred at room temperature for 16 h. Potassium carbonate solution (20mL) was added to dilute the reaction and extracted with DCM (2X 5 mL). The combined organic extracts were concentrated in vacuo to give a crude residue. Purification by column chromatography on silica gel using ethyl acetate: methanol: 0.88 ammonia (gradient 98: 2: 0.2 to 90: 10: 1) gave 221mg (86%) of the title compound as a cream-colored foam.

¹H NMR(400MHz，CD₃OD)δ1.30-1.40(1H，m)，1.45-1.60(1H，m)，1.65-1.80(1H，m)，1.80-1.90(1H，m)，2.70-2.90(2H，m)，2.95-3.15(1H，m)，3.40-3.60(4H，m)，3.64(3H，s)，3.65-3.75(1H，m)，3.80-4.00(4H，m)，4.05-4.25(1H，m)，6.85-7.00(3H，m)，7.10-7.20(2H，m)，7.20-7.30(3H，m)，7.40-7.55(1H，m)。LRMS：EI⁺m/z 542[MH⁺]。

Preparation 67-71

These compounds were prepared by the method of preparation 66 using the appropriate precursors listed in the table below.

A is a mixture of epimers; b is a single epimer

Preparation of 72-85

These compounds were prepared by the method of example 66 using the appropriate precursors listed in the table below. In some cases, single epimers (marked with an asterisk) were separated from the initially produced epimer mixture by normal phase chromatography.

A is a mixture of epimers; b is a single epimer; a single epimer with an unknown absolute configuration at the C ═ phenyl substitution site; single epimer with the opposite configuration at the chloro-phenyl substitution site relative to preparation 73 (respectively, relative to preparation 84)

Preparation of 86-91

These compounds were prepared by the method of example 98 using the appropriate precursors listed in the table below.

A is a mixture of epimers; b is a single epimer

Preparation 92: 5- (4-chlorophenyl) -1- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-3-yl]Carbonyls 4- (methylsulfonyl) -1, 4-diazacyclooctane radical

Diethylisopropylamine (0.298mL, 1.71mmol) was added to a solution of preparation 83(239mg, 0.389mmol) in DCM (10 mL). 1-Chloroethyl chloroformate (0.340mL, 3.12mmol) was added and the solution was stirred at reflux for 20 h. The reaction was concentrated in vacuo and 10% citric acid solution (10mL) was added to dilute the residue and extracted with DCM (3 × 15 mL). The combined organic extracts were concentrated in vacuo and the residue was dissolved in methanol (10mL) and stirred at reflux for 2 h. The reaction mixture was concentrated in vacuo to give a crude residue. Purification by column chromatography on silica gel using dichloromethane: methanol: 0.880 ammonia (90: 10: 1) gave 270mg (100%) of the title compound as a brown oil (as a mixture of the two epimers).

¹H NMR(400MHz，CD₃OD)δ1.32-1.67(2H，m)，2.37-2.50(3H，m)，2.81-3.25(2H，m)，3.27-3.31(3H，m)，3.32-3.94(12H，m)，4.76-4.87(1H，m)，6.53-6.72(2H，m)，6.94-6.99(1H，m)，7.11-7.16(1H，m)，7.19-7.36(3H，m)。LRMS：APCI⁺m/z 524[MH⁺]。

Preparation 93 to 97

These compounds were prepared by the method of preparation 92 using the appropriate precursors listed in the table below.

A is a mixture of epimers; b is a single enantiomer; a single epimer with an unknown absolute configuration at the C ═ chloro-phenyl substitution site; d-a single epimer having the opposite configuration at the chlorophenyl substitution site relative to the mixture of preparation 94; e ═ racemate

Preparation 98-108

These mixtures are described in the patent documents listed in the following table.

Biological data

Data for representative compounds of the invention are given below.

Not determined

Claims (8)

1. A compound selected from:

6- [ (3S,4R) -3- { [5S- (4-chlorophenyl) -4- (3,3, 3-trifluoropropionyl) -1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-1-yl ] -2-methylpyridazin-3 (2H) -one;

6- [ (3S,4R) -3- { [5S- (4-chlorophenyl) -4-isobutyryl-1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (2, 4-difluorophenyl) pyrrolidin-1-yl ] -2-methylpyridazin-3 (2H) -one;

6- [ (3S,4S) -3- { [5S- (4-chlorophenyl) -4-isobutyryl-1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (5-chloropyridin-2-yl) pyrrolidin-1-yl ] -2-methylpyridazin-3 (2H) -one;

8- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

8S- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

8R- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

8R- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2-fluoro-4-methoxyphenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

6- [ (3S,4R) -3- { [ 4-acetyl-5S- (4-chlorophenyl) -1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (2-fluoro-4-methoxyphenyl) pyrrolidin-1-yl ] pyridazin-3 (2H) -one;

8S- (4-chlorophenyl) -4- { [ (3S,4S) -4- (5-chloropyridin-2-yl) -1- (6-cyanopyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

1- { [ (3S,4S) -1-tert-butyl-4- (5-chloropyridin-2-yl) pyrrolidin-3-yl ] carbonyl } -5S- (4-chlorophenyl) -4-isobutyryl-1, 4-diazacyclooctane;

6- [ (3S,4S) -3- { [ 4-acetyl-5S- (4-chlorophenyl) -1, 4-diazacyclooctan-1-yl ] carbonyl } -4- (5-chloropyridin-2-yl) pyrrolidin-1-yl ] [1,2,4] triazolo [4,3-b ] pyridazine;

8S- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2, 4-difluorophenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylic acid methyl ester;

and pharmaceutically acceptable salts thereof.

2. A compound which is methyl 8S- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2, 4-difluorophenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylate, or a pharmaceutically acceptable salt thereof.

3. A pharmaceutical composition comprising a compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or adjuvant.

4. A pharmaceutical composition comprising a compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or adjuvant.

5. A pharmaceutical composition comprising methyl 8S- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2, 4-difluorophenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylate or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or adjuvant.

6. A pharmaceutical composition comprising methyl 8S- (4-chlorophenyl) -4- { [ (3S,4R) -4- (2, 4-difluorophenyl) -1- (1-methyl-6-oxo-1, 6-dihydropyridazin-3-yl) pyrrolidin-3-yl ] carbonyl } -1, 4-diazacyclooctane-1-carboxylate or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable diluent or adjuvant.

7. Use of a compound according to any one of claims 1 to 2, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for a condition which is benefited by agonism of the MC4 receptor.

8. Use according to claim 7, wherein the disorder is male or female sexual dysfunction, obesity, diabetes or a urological disorder.