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WO1996040781A1 - INHIBITEURS PEPTIDIQUES CYCLIQUES D'ADHERENCE CELLULAIRE INDUITE PAR INTEGRINE β1 ET β¿2? - Google Patents

INHIBITEURS PEPTIDIQUES CYCLIQUES D'ADHERENCE CELLULAIRE INDUITE PAR INTEGRINE β1 ET β¿2? Download PDF

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Publication number
WO1996040781A1
WO1996040781A1 PCT/US1996/010186 US9610186W WO9640781A1 WO 1996040781 A1 WO1996040781 A1 WO 1996040781A1 US 9610186 W US9610186 W US 9610186W WO 9640781 A1 WO9640781 A1 WO 9640781A1
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Prior art keywords
group
cys
compound
asp
fca
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PCT/US1996/010186
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English (en)
Inventor
Thomas J. Lobl
Shiu-Lan Chiang
Wolfgang Scholz
Nancy Delaet
Pina Cardarelli
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Mitsubishi Tanabe Pharma Corp
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Tanabe Seiyaku Co Ltd
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Priority to AU63321/96A priority Critical patent/AU6332196A/en
Priority to US08/738,838 priority patent/US5821329A/en
Priority claimed from US08/738,838 external-priority patent/US5821329A/en
Publication of WO1996040781A1 publication Critical patent/WO1996040781A1/fr
Anticipated expiration legal-status Critical
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K7/00—Peptides having 5 to 20 amino acids in a fully defined sequence; Derivatives thereof
    • C07K7/02—Linear peptides containing at least one abnormal peptide link
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to novel cyclic peptides and peptidomimetic compounds which are characterized by cell adhesion modulation activity.
  • Vascular endothelial cells form the interface between blood and tissues and control the passage of leukocytes as well as plasma fluid into tissues.
  • a variety of signals generated at the site of inflammation can activate both endothelial cells as well as circulating leukocytes so that they become more adhesive to one another.
  • Activation of endothelium and leukocytes initiates a complex adhesion cascade.
  • This adhesion cascade involves the "tethering" of the leukocytes to the endothelium, after which they "roll” along the endothelial surface and finally strongly adhere and migrate into tissue to perform host defense functions.
  • integrins Several adhesion molecules, belonging to a super gene family consisting of non-covalently associated heterodimeric proteins called integrins, have been identified as being involved in leukocyte- endothelial cell interactions.
  • the ⁇ 2 integrin subfamily includes LFA-1 (CDlla/CD18) , Mac-1 (CDllb/CD18, CR3) and pl50/95 (CDllc/CD18, CR4) .
  • LFA-1 CDlla/CD18
  • Mac-1 CDllb/CD18, CR3
  • pl50/95 CDllc/CD18, CR4
  • the known ligands for LFA-1 are ICAM-1, ICAM-2 and ICAM-3.
  • the Intracellular Adhesion Molecules (ICAM) are members of the Ig gene superfamily.
  • ICAM-1 is the most ubiquitous of the ICAMs, being expressed in low levels on most peripheral blood leukocytes as well as endothelial cells, fibroblasts and dendritic cells. Cytokine activation of endothelial cells induces a dramatic increase in the expression of ICAM-1 and LFA-l/lCAM-1 interactions which are integral to both lymphocyte adhesion and transmigration through the endothelial barrier; Dustin, M. . et al, J. Immunol. , 137, 245-254 (1986) . ICAM-2 is primarily constitutively expressed on endothelial cells; de Fougerolles, A.R. et al, J. EXP. Med..
  • ICAM-3 is largely found on resting lymphocytes, monocytes and neutrophils; and shows increased expression upon T cell activation; de Fougerolles, A.R. and Springer, T.A. , J. EXP. Med.. 175. 185-190 (1992) .
  • ICAM-1 In addition to its critical role in mediating cellular adhesion, ICAM-1 has also been shown to act as a receptor for a subgroup of rhinoviruses known as the major groups, and soluble ICAM-1 has been shown to act as specific inhibitor of rhinovirus infection; Martin, S.D. et al, Nature. 344. 70-72 (1990) .
  • a compound which blocks the interaction of rhinovirus with ICAM-1 may be a powerful pharmacological agent for the prevention and treatment of colds and secondary complications arising from rhinovirus infection.
  • ⁇ integrin subfamily include a ⁇ i > oe 2 ⁇ l l a 2 ⁇ l t a ⁇ ⁇ (V A-4, CD49d/CD29) , ot ⁇ ⁇ , ot 5 ⁇ 1 (V A-5, CD49e/CD29) , a 6 ⁇ l t a e ⁇ ⁇ and v ⁇ ⁇ (CD51/CD29) .
  • a ⁇ binds to both fibronectin and the cytokine inducible molecule on endothelial cells termed vascular cell adhesion molecule (VCAM) . interaction has been shown to be involved with lymphocyte extravasation at sites of chronic inflammation; Elices, -M.J. et al, Cell. 6>fJ, 577- 584 (1990) .
  • MadCAM macosal addressin cell adhesion molecule
  • ⁇ 2 integrins and ICAM have been shown to have important roles in graft rejection, ischemia reperfusion, delayed type hypersensitivity, asthma and allergies, inflammatory bowel disease, rheumatoid arthritis and AIDS.
  • Support for the importance of ⁇ 2 integrins in mediating inflammatory responses has been demonstrated by the ability of monoclonal antibodies which recognize LFA-1 to block CTL-mediated lysis of target cells, as well as inhibit proliferation of T cells in response to soluble antigens, alloantigens and mitogen.
  • Several in vivo models have demonstrated the importance of ⁇ 2 integrins in delayed-type hypersensitivity.
  • Anti-LFA-1 antibodies have been shown to block the migration of spleen T cells to sites of dermal inflammation, as well as the homing of lymph node and spleen T cells to peripheral and mesenteric lymph node in rats; Issekutz, T.B., J. Immunol.. 149. 3394- 3402 (1992) .
  • Both anti-LFA-1 and anti-ICAM-1 antibodies can reduce ear swelling caused by edema and cell infiltration in association with delayed-type hypersensitivity; Scheynius, A. et al, J. Immunol.. 150. 655-663 (1993) .
  • Anti-ICAM-1 antibodies have also been shown to attenuate airway eosinophilia, hyperresponsiveness and asthma symptoms in a primate asthma model.
  • One objective of the present invention is to provide compounds which act to modulate cell adhesion. These compounds are represented by the formula (I) :
  • L 1 is a residue of an amino acid or an amino acid mimetic, having a functional group suitable for the formation of a cyclizing bridge, Z, between L 1 and L 2 .
  • L 2 is a residue of an amino acid or an amino acid mimetic, having a functional group suitable for the formation of a cyclizing bridge, Z, between L 1 and L 2 .
  • Z is a cyclizing moiety or bond between L 1 and L 2 ;
  • X° is a residue of an amino acid or an amino acid mimetic and r is an integer of 0 or 1.
  • X 1 is a residue of an amino acid or an amino acid mimetic and m is an integer of 0, 1 or 2.
  • X 2 is jS-Asp or ⁇ -Glu or an ester or an amide derivative formed from the -carboxyl group of any of the aforementioned groups.
  • X 3 is Ser(R 4 ), Thr(R 4 ), Tyr(R 4 ), Ala, Gly, Lys(R 4 ), Orn(R 4 ), Dpr(R 4 ), N-Me-Ala, Aib, Val, Tic, o- or m- halo-Tyr, dihalo-Tyr, p-halo- Phe, dihalo-Phe, Sar, Leu, lie, Nle or Cys(R 4 ) wherein when X 3 is Ser(R 4 ), Thr(R 4 ) or Tyr(R 4 ), then R 4 is a substituent of the side chain hydroxyl group of X 3 and is selected from the group consisting of a hydrogen atom, a lower alkyl, a cycloalkyi, a cycloal
  • q is an integer of 0 or 1.
  • X 5 is a residue of an amino acid or an amino acid mimetic, n is an integer of 0, 1, 2 or 3.
  • X 6 is a residue of an amino acid or an amino acid mimetic, s is an integer of 0 or 1.
  • R 1 is a hydrogen atom, Xan, Fmoc, 9-FAc, 9-FCA, 1- FCA, Ac, 2-NaphAc, Ada, 5-Fine, biotinyl, Su, 1-NaphCA, For, 1-NaphAc, PhAc, l-NaphS0 2 , 2-NaphS0 2 or 2-NaphCA, with the proviso that the compound (Fmoc) -Arg-Cys*- ( ⁇ - Asp) - (ThioP) -Cys* is not included.
  • the present invention includes compounds having an IC 50 of less than about 500 ⁇ M as established in JY-Endothelial cell and Jurkat-Endothelial cell adhesion assays; in another regard, the invention includes compounds having an IC 50 of less than about 100 ⁇ M in such assays.
  • the invention also includes methods for obtaining (either in vi tro or in vivo) , such integrin receptor adhesion inhibition.
  • the compounds of the present invention accomplish strong inhibition, at low concentrations, with an IC 50 of less than about 500 ⁇ M, or alternatively less than about 100 ⁇ M.
  • the present invention includes compounds having an IC 50 of less than about 200 ⁇ M as established in Jurkat-endothelial cell and JY-endothelial cell adhesion assays. Compounds with activity below 100 ⁇ M are most preferred, below 150 ⁇ M are preferred, below 500 ⁇ M are less preferred and above 500 ⁇ M least preferred.
  • the invention also includes methods for obtaining (either in vitro o ⁇ in vivo) such leukocyte receptor adhesion inhibition.
  • the compounds of the present invention accomplish strong inhibition at low concentrations, with an IC 50 of less than about 250 ⁇ M.
  • Another objective of the present invention is to provide compounds, pharmaceutical compositions, and methods which may be used in the study, diagnosis, treatment or prevention of diseases and conditions which involve or relate to cell adhesion.
  • Another objective of the present invention is to provide compounds, pharmaceutical compositions and methods of treatment which may be used to inhibit inflammatory diseases in humans.
  • Another objective is to provide antibodies made by using the compounds of the invention, such as, but not limited to, antibodies to the compounds and anti- idiotype antibodies to the antibodies disclosed in order to study, diagnose, treat or prevent the above mentioned diseases and conditions which relate to cell adhesion.
  • Another objective of the present invention is to provide a matrix which can be used to purify proteins, polysaccharides or other compounds which specifically bind to the cyclic peptides of the present invention with high affinity.
  • the present invention relates to compounds meeting one or more of the above objectives.
  • the compounds have activity as cell adhesion modulators.
  • the novel compounds have been found to modulate cell adhesion by competing, for example, with integrin specific ligands, such as but not limited to, ICAM-1 and VCAM, which are sufficiently inhibited from binding to the leukocyte receptors so as to prevent or reduce cell-cell adhesion.
  • integrin specific ligands such as but not limited to, ICAM-1 and VCAM
  • Other uses for the compounds include enhancing cell adhesion to a distinct surface. These useful compounds can function as both cell-adhesion agonists and antagonists.
  • Cell adhesion is required for certain normal physiological functions. Yet, there are situations in which cell adhesion is undesirable, or in which modulated cell adhesion is desirable. Many pathologies exist which are related to abnormal cellular adhesion. These include, but are not limited to, rheumatoid arthritis, asthma and allergies, adult respiratory distress syndrome (ARDS) , AIDS, cardiovascular disease thrombosis or harmful platelet aggregation, reocclusion following thrombolysis, allograft rejection, reperfusion injury, psoriasis, eczema, contact dermatitis and other skin inflammatory diseases, osteoporosis, osteoarthritis, atherosclerosis, neoplastic disease including metastasis of neoplastic or cancerous growth, treatment of certain eye diseases such as detaching retina, Type I diabetes, multiple sclerosis, systemic lupus erythematosus (SLE) , inflammatory and immunoinflammatory conditions including ophthalmic inflammatory conditions and inflammatory bowel disease (
  • a peptide or other compound with suitable affinity for integrin receptors when attached, for example, to a suitably positioned matrix or surface, may be able to promote beneficial cell adhesion and resultant enhancement of wound healing.
  • ICAM and VCAM expression can be seen at post-capillary venuole sites. This increased expression has been suggested to play a role in increased risk of arterial blockage from increased cellular adhesion at these sites. Compounds which effect ICAM and VCAM interactions with integrins should be useful in treatment of this type of arterial blockage.
  • Anti-ICAM antibodies are immunosuppressive in monkeys and aid in preventing graft rejection (Wee et al. Trans. Proc.. 23. 279 (1991)) .
  • Compounds, such as the ones encompassed by this invention are expected to be useful as immunosuppressives for preventing graft rejection. They should also have the advantage over antibodies of being less antigenic in vivo because of their small size and structural design.
  • cell adhesion modulation compounds of the present invention are represented in part by amino acid sequence formulas wherein the individual amino acids are represented by their standard three-letter abbreviations, or alternatively, they are represented by one-letter abbreviations as indicated in the following Table of Abbreviations. Table of Abbreviations
  • Amino acids of the peptide backbone are delineated with hyphens; synthetic amino acids, unnatural amino acids, and amino acid mimetics are enclosed in parentheses; the R 1 -R 5 groups which are attached to the backbone amino acid groups are enclosed in parentheses and are immediately adjacent to the amino acid to which they are attached without an intervening hyphen. Additional abbreviations used herein include the following:
  • Aib ⁇ -aminoisobutyric acid j ⁇ -Ala: j ⁇ -alanine (3-aminopropionic acid) jS-Asp: j ⁇ -aspartic acid
  • BCECF-AM 2' ,7'-bis- (2-carboxyethyl) -5- (and -6)- carboxyfluorescein, acetoxymethyl ester, Molecular Probes Cat. No. B-1170
  • BSA bovine serum albumin Cha: 3-Cyclohexyl-2-aminopropionic acid
  • CTL cytotoxic T-lymphocyte
  • DCM dichloromethane
  • DIEA diisopropylethylamine
  • Fmoc 9-Fluorenylmethoxycarbonyl FN: fibronectin
  • HSA human serum albumin
  • HUVEC human umbilical vein endothelial cell
  • ICAM intercellular adhesion molecule
  • IC 50 inhibitory concentration, concentration at which adhesion is inhibited to 50% of control level
  • IL-1 interleukin 1
  • LFA-1 Lymphocyte Functional Antigen-1
  • 2-NaphCA 2-Naphthalenecarbonyl l-NaphS0 2 : 1-Naphthalenesulfonyl
  • PBS phosphate buffered saline
  • Pen penicillamine
  • PhAc Phenylacetyl
  • R.T. room temperature (about 24°C)
  • rTNF recombinant Tumor Necrosis Factor
  • Thr(OMe) O-methylthreonine
  • Tic l,2,3,4-tetrahydroisoquinoline-3- carboxylic acid
  • TNF Tumor Necrosis Factor
  • TTC Tetrahydro-1,3-thiazine-4-carboxylic acid
  • Tyr(OMe) O-methyltyrosine
  • VCAM vascular cell adhesion molecule
  • VLA very late activation antigens
  • R 1 is a substituent on the - amino terminal group of the N-terminal amino acid.
  • R 1 is preferably a hydrogen atom, Ada, 1- or 2-NaphAc, 1- NaphS0 2 , 2-NaphS0 2 , 1-NaphCA, 2-NaphCA, 9-FAc, 1-FCA, 9-
  • FCA Chloroc
  • PhAc PhAc
  • Ac PhAc
  • Fmoc Fmoc
  • Xan 5-Fine
  • biotinyl Su and For.
  • X° is an optional group, r is an integer of 0 or 1, and when present, X° is preferably
  • Trp 1-Nal, 2-Nal, or an aromatic or hydrophobic amino acid, such as Tyr, Leu, Nle, He, Val, NorVal and Cha.
  • X 1 is an optional group, "m” is an integer of 0, 1 or 2 and when present, X 1 is preferably a hydrophobic amino acid or a side chain-protected basic amino acid residue having a side chain terminal amino group with the formula -NHR 2 , where R 2 is selected from the group consisting of a hydrogen atom, PhAc, 1-NaphAc, 2-NaphAc, Ada, 2-norbornylacetyl, 1-FCA, CHAc and For.
  • Preferred moieties for X 1 include Arg, Lys(R 2 ), Gly, N ⁇ - Me-Arg, Nle, Cha, Orn(R 2 ) , Val, NorVal, Dpr(R 2 ), Abu, Leu and He.
  • X 2 is selected from D- and L-/3-Asp and D- and L- ⁇ -Glu and ⁇ -carboxyl esters or amides of any of these aforementioned amino acids.
  • the carboxyl peptide linkages of jS-Asp and ⁇ -Glu are formed between the side chain carboxyl groups ( ⁇ -carboxyl or ⁇ - carboxyl) of jS-Asp and ⁇ -Glu, respectively, and the a- amino group of the adjacent amino acid.
  • the ⁇ -carboxyl group of X 2 may have the formula -C(0)R 3 , where R 3 is -OR, -NHR or -NRR and where R is a hydrogen atom, a lower alkyl, an aryl, an arylalkyi, a cycloalkyi, a heteroaryl-lower alkyl or a heterocyclic group as appropriate for an amide or ester. More specifically, R may be methyl, Fm, benzyl, cyclohexyl, 1-adamantyl, 2-, 3- or 4-picolyl, 1- or 2-naphthylmethyl or isobutyl.
  • R 3 is -OR and R is a hydrogen atom, -C(0)OH is present.
  • R 3 is -NHR or -NRR and R is a hydrogen atom, a carboxamide is present. Otherwise, for -NRR each R group may be selected independently of the other R group.
  • X 3 is an optional amino acid residue where "p" is an integer of 0 or 1.
  • preferred amino acids include Ser(R 4 ) , Thr(R 4 ) , Tyr(R 4 ), Ala, Gly, Lys(R 4 ), Orn(R 4 ), Dpr(R 4 ), N-Me-Ala, Aib, Val, Tic, o- and m-halo-Tyr, dihalo-Tyr, p-halo- Phe, dihalo-Phe, Sar, Leu, He, Nle and Cys(R 4 ) .
  • X 4 is an optional amino acid, where "q" is an integer of 0 or 1.
  • preferred amino acids include Pro, ThioP, Aib, Asn, TCA, TTC, Sar, N-Me-Ala, other N-methylated natural amino acids, Tic and pipecolinic acid (homoproline) .
  • X 5 is optional and "n” is an integer of 0, 1, 2 or 3.
  • X 6 is optional and "s" is an integer of 0 or 1.
  • Preferred groups for X 5 and/or X 6 include a side chain-protected derivative of a basic amino acid residue having a side chain terminal amino group with the formula -NHR 4 ; a side chain-protected derivative of an acidic amino acid residue having a side chain terminal carboxyl group with the formula -C(0)R 4 , where R 4 is -OR, -NHR or -NRR; Ser(R 4 ) , Thr(R 4 ) , Tyr(R 4 ) , and R 4 is a substituent of the hydroxyl group; and Cys(R 4 ) and R 4 is a substituent of the sulfhydryl group.
  • X 5 and/or X 6 include Ala; Asp(R 4 ) ; Glu(R 4 ) ; Ser(OMe) ; Thr(OMe) ; Tyr(OMe) ; Gly; Abu; AnC; Leu; Val; ⁇ -Ala; Lys(R 4 ); Dpr(R 4 ); Orn(R 4 ), He and Nle.
  • R 4 is a substituent of a terminal amino group of a side chain of a basic amino acid, of a ⁇ - or ⁇ -carboxylic group of an acidic amino acid, of a side chain hydroxyl group of Ser, Thr, Tyr or of a hydroxyl containing derivative or analog of Ser, Thr or Tyr or of a sulfhydryl group of Cys or of a sulfhydryl containing derivative or analog of Cys .
  • the terminus of the side chain may be represented by the formula -C(0)R 4 and R 4 is -OR, -NHR or -NRR, wherein R may be a hydrogen atom, a lower alkyl, an aryl, an arylalkyi, a cycloalkyi, a heteroaryl-lower alkyl or a heterocyclic group as is appropriate for an amide or ester functional group. More specifically, R may be benzyl, cyclohexyl, 1- or 2- naphthylmethyl, 2-, 3- or 4-picolyl, 1-adamantyl, Fm, isobutyl or methyl.
  • R 4 is -OR and R is H, the side chain terminal functionality -C(0)OH, is present.
  • R 4 is NHR or NRR and R is H, a carboxamide is present.
  • each R group may be selected independently of the other R group.
  • R 4 is a substituent of the hydroxyl group and may be a hydrogen atom, a lower alkyl, a cycloalkyi, a cycloalkyi-lower alkanoyl, an aryl, an arylalkyi, an aryloxycarbonyl, an arylcarbonyl, a heteroaryl-lower alkyl, an arylacetyl, a heterocyclic group, a lower alkanoyl, or an arylalkoxycarbonyl.
  • R 4 may be methyl, benzyl, cyclohexyl, 1- or 2- naphthylmethyl, Fm, 2-, 3- or 4-picolyl or isobutyl. If R 4 is a hydrogen atom, a hydroxyl side chain is present.
  • R 4 When X 3 , X 5 or X 6 is a residue of a basic amino acid, the terminal group of the side chain may be denoted by the formula -NHR 4 .
  • Preferred groups for R 4 include a hydrogen atom, lower alkyl, aryl, aryloxycarbonyl, arylcarbonyl, arylacetyl, an arylalkyi, an arylalkoxycarbonyl, a lower alkanoyl, cycloalkyi, and cycloalkyi-lower alkanoyl. More specifically, R 4 may be cyclohexyl, 1- and 2-naphthylmethyl, Fmoc, 1-FCA, 9-FCA, Ada, Ac and 9-FAc. When R 4 is a hydrogen atom, an amino terminus of the side chain is present.
  • R 4 is a substituent of the sulfhydryl group and is a hydrogen atom, a lower alkyl, an arylalkyi, an aryl, a cycloalkyi, a cycloalkyi-lower alkanoyl, an aryloxycarbonyl, an arylcarbonyl, an acetamido-lower alkyl, an arylacetyl, a heterocyclic group, a lower alkanoyl, or an arylalkoxycarbonyl.
  • R 4 is preferably methyl, benzyl, 1- or 2-naphthylmethyl, cyclohexyl, Fm, Acm, 4-methylbenzyl or Ada.
  • R 4 is preferably not a hydrogen atom; however, where R 4 is a hydrogen atom, a sulfhydryl terminal group of the side chain is present.
  • the ⁇ -carboxyl group may be represented by the formula -C(0)R 5 , where R 5 is -OR, -NHR, or -NRR and R is a hydrogen atom, a lower alkyl, an arylalkyi, an aryl, a cycloalkyi, a heteroaryl-lower alkyl or a heterocyclic group as is appropriate for an amide or ester functional group. More specifically, R may be methyl, Fm, isobutyl, benzyl, cyclohexyl, 1- or 2-naphthylmethyl, 1- adamantyl or 2-, 3- or 4-picolyl.
  • R 5 is -OR and R is H, an ⁇ -carboxyl terminus, -C(0)OH, is present.
  • R 5 is -NHR or -NRR and R is H, a carboxamide is the C- terminus of the peptide.
  • each R group may be selected independently of the other R group.
  • R 1 -R 5 are preferably each independently selected so as to enhance the specific activity of the resulting compound and/or to preserve the compound against metabolism in the in vivo environment and to increase the effective half-life of the compound.
  • the use of one or more groups for X°-X 6 particularly at the N-terminal position or in other regions in the compound, promotes the activity of the compound.
  • a bridge is formed via a cyclizing moiety, Z, between L 1 and L 2 .
  • L 1 and L 2 are each, or together, residues of amino acids or amino acid mimetics having functional groups suitable for the formation of a cyclizing bridge between L 1 and L 2 .
  • L 1 and L 2 are chosen so that each contains a functional group which contributes to the formation of the cyclizing bridge moiety, Z.
  • Z is formed from functional groups contributed by L 1 and L 2 and may also contain additional atoms and spacer groups.
  • preferred functional groups include thiol, amino and carboxyl groups. Such functional groups may be borne on the side chain of amino acids or amino acid mimetics, or may constitute the ⁇ -amino group (in L 1 ) or terminal-carboxyl group (in L 2 ) thereof.
  • L 1 and L 2 are Cys, Cys-NH 2
  • L 1 and L 2 are each selected from Cys or Pen. Both Cys and Pen contain a sulfhydryl group and thus, the bridging cyclization can be accomplished by oxidative coupling of the sulfhydryls to form a disulfide bond between residues L 1 and L 2 .
  • the cyclizing moiety, Z is a covalent bond between the two sulfur atoms. This may be depicted generally for compounds wherein, both L 1 and L 2 are Cys residues as follows:
  • L 1 is Cys and L 2 is Cys.
  • the cyclizing moiety may also be formed by a hydrocarbon moiety, for example a (poly)methylene bridge moiety of the form - (CH 2 ) - where y is an integer from
  • L 1 and L 2 may be chosen from other natural or synthetic, - or D-amino acids or amino acid mimetics which provide a side chain or an amino- or carboxyl-terminus suitable as a functional group, for the formation of a cyclizing moiety.
  • L 2 may be selected from Asp, Glu, or other natural or synthetic amino acids which provide a suitable side chain carboxyl group for cyclic linkage, through formation of an amide bond in a condensation reaction, with an amino group (e.g., an N ⁇ -amino group, or an ⁇ side chain amino group as on, for example, Lys or Orn) on L 1 .
  • the cyclizing moiety Z may be a simple amide bond between L 1 and L 2 .
  • an amino acid residue L 2 may provide an ⁇ - carboxyl group from its carboxyl terminus for amide linkage with either a side chain amino or ⁇ -amino group on L 1 ; the direction of the amide linkage may also be reversed where L 1 provides a side chain carboxyl group and L 2 provides a side chain amino group.
  • the cyclizing bridge between L 1 and L 2 may also be formed via a monosulfide (thioether) linkage, as exemplified below.
  • thioether monosulfide
  • the ⁇ -carboxyl group may be represented as -C(0)R 5 where R 5 is -OR, -NHR or -NRR and R is H, a lower alkyl, an aryl, an arylalkyi, a cycloalkyi, a heteroaryl-lower alkyl or a heterocyclic group as is appropriate for an amide or ester functional group.
  • R may be methyl, benzyl, cyclohexyl, 1- or 2-naphthylmethyl, 1-adamantyl, Fm, 2-, 3- or 4- picolyl or isobutyl.
  • R 5 is -OR and R is H, the ⁇ - carboxyl group, -C(0)OH, is present on L 2 .
  • R 5 is - NHR or -NRR and R is H, the carboxamide is present.
  • each R group in -NRR may be selected independently of the other R group.
  • Synthetic amino acid residues may also be utilized for L 1 or L 2 , as for example, homologues wherein a side chain is lengthened or shortened while still providing the sulfhydryl carboxyl, amino or other reactive functional group for cyclization.
  • the preferable compounds are compounds of the formula (I) wherein L 1 and L 2 are each or together selected from the group consisting of Cys, Cys-NH 2 (only for L 2 and when no X 5 or X 6 groups are present) , Arg (only for L 1 and when X° and X 1 are absent) Pen, Orn, Lys, Dpr, Gly (only for L 1 and when X° and X 1 are absent) , Met, Glu and Asp;
  • Z is selected from a single bond, a lower alkylene or formula: -C0CH 2 -;
  • X° is selected from the group consisting of Trp, 1- Nal, 2-Nal, Tyr, Leu, Nle, He, Val, NorVal and Cha;
  • X 1 is selected from the group consisting of Lys(R 2 ) , Gly, N ⁇ -Me-Arg, Nle, Cha, Val, NorVal, Dpr(R 2 ), Arg, Orn(R 2 ), Leu, He and Abu;
  • R 2 is selected from a hydrogen atom, Ada, 2- norbornylacetyl, 1-FCA, 1-NaphAc, 2-NaphAc, PhAc, For or CHAc;
  • X 2 is selected from the group consisting of ⁇ -Asp and j ⁇ -Asp(R 3 ), ⁇ -Glu and ⁇ -Glu(R 3 ) and R 3 is -OR, -NHR or -NRR and where R is a hydrogen atom, a lower alkyl, an aryl, an arylalkyi, a cycloalkyi, a heteroaryl-lower alkyl or a heterocyclic group; and
  • X 5 and/or X 6 is selected from the group consisting of Ala; Asp(R 4 ) and Glu(R 4 ) wherein R 4 is a substituent of the derivatized side chain terminal carboxyl group and is selected from the group consisting of -OR, -NRR and -NHR where R is selected from the group consisting of a hydrogen atom, a lower alkyl, an aryl, a cycloalkyi, a heteroaryl-lower alkyl, a heterocyclic group and an arylalkyi; Ser(R 4 ) , Tyr(R 4 ) and Thr(R 4 ) wherein R 4 is OR and R is selected from the group consisting of a hydrogen atom, a lower alkyl, a cycloalkyi, an aryl, an arylalkyi, a cycloalkyi-lower alkanoyl, an aryloxycarbonyl, an arylcarbonyl, a heteroary
  • R 4 is a substituent of the derivatized side chain terminal amino group and is selected from the group consisting of a hydrogen atom, a lower alkyl, a cycloalkyi, a lower alkanoyl, an arylalkyi, an arylalkoxycarbonyl, an aryl, an aryloxycarbonyl, an arylcarbonyl, a cycloalkyi-lower alkanoyl and an arylacetyl; AnC; Leu; Val; He; Nle; ⁇ - Ala and Cys(R 4 ) wherein R 4 is a substituent of the derivatized side chain sulfhydryl group expressed as -SR and R is selected from the group consisting of a hydrogen atom, a lower alkyl, a cycloalkyi, an acetamido-lower alkyl,
  • R 1 is selected from the group consisting of hydrogen, Xan, 9-FAc, 9-FCA, 1-
  • FCA Ac, 2-NaphAc, Ada, 5-Finc, biotinyl, Su, 1-NaphCA, For, 1-NaphAc, PhAc, l-NaphS0 2 , 2-NaphS0 2 and 2-NaphCA.
  • most preferable ones are those wherein p is an integer of 1.
  • R 1 is selected from hydrogen, 1-FCA, 9-FAc, Ada, 5- Finc, Ac, Biotinyl, 9-FCA, Fmoc, For, 1- or 2-NaphAc, PhAc, l-NaphS0 2 , 2-NaphS0 2 , 1- or 2-NaphCA, Xan or Su;
  • X° is selected from the group consisting of Trp, 1- Nal, 2-Nal, Tyr, Leu, Nle, He, Val, NorVal and Cha;
  • r is an integer of 0 or 1;
  • X 1 is selected from Arg, N ⁇ -Me-Arg, Lys, Lys (Ada) , Lys (1-NaphAC) , Lys (PhAc), Lys(For), Lys (2-NaphAc) , Orn, Orn(PhAc), Gly, Nle or Cha; m is an integer of 0 or 1;
  • L 1 is selected from Cys, Lys, Pen, Orn or Dpr;
  • X 2 is selected from 3-Asp or ⁇ -Asp( ⁇ OFm) ;
  • X 3 is selected from Ser, N-Me-Ala, Gly or Ala;
  • X 4 is selected from ThioP, Asn, TCA, Pro, Aib or Tic;
  • L 2 is selected from Cys, Cys-NH 2 (only when no X 5 or X 6 groups are present) , Asp, Pen or Glu;
  • X 5 is selected from Ala, Gly, Abu or Lys(Fmoc) ;
  • X 6 is selected from Asp- (OFm) , Asp- (NH-Ada) , Ser, Lys, Lys(Fmoc) -NH 2 or Gly.
  • R 1 is selected from a hydrogen atom, l-FCA, 9-FAc, Ada, 5-Fine, Ac, Biotinyl, 9-FCA, For, 9-FAc, 2-NaphAc, PhAc, l-NaphS0 2 , 2-NaphS0 2 , 2-NaphCA or Xan.
  • Z is selected from a single bond, a lower alkylene or a group of the formula: -COCH 2 -;
  • R 1 is selected from hydrogen, l-FCA, 9-FAc, Ada, 5- Fine, Ac, Biotinyl, 9-FCA, Fmoc, For, 9-FAc, 2-NaphAc, PhAc, 2-NaphCA or Xan;
  • X° is Trp; r is an integer of 0 or 1;
  • X 1 is selected from Arg, N ⁇ -Me-Arg, Lys, Lys(Ada), Lys(1-NaphAc) , Lys(PhAc) , Lys(For), Lys(2-NaphAc) , Orn, Orn(PhAc), Gly, Nle or Cha; m is an integer of 0 or 1;
  • L 1 is selected from Cys, Lys or Pen;
  • X 3 is selected from Gly or Ala;
  • X 4 is selected from ThioP, Asn, Pro or Tic;
  • L 2 is selected from Cys, Cys-NH 2 (only when no X 5 or X 6 groups are present) , Asp or Pen;
  • X 5 is selected from Ala, Gly, Abu or Lys(Fmoc) ; and X 6 is selected from Asp- (OFm) , Asp- (NH-Ada) , Ser, Lys, Lys(Fmoc) -NH 2 or Gly.
  • X 6 is selected from Asp- (OFm) , Asp- (NH-Ada) , Ser, Lys, Lys(Fmoc) -NH 2 or Gly.
  • R 1 is selected from a hydrogen atom, l-FCA, 9-FAc, Ada, 5-Finc, Ac, Biotinyl, 9-FCA, For, 9-FAc, 2-NaphAc, PhAc, l-NaphS0 2 , 2-NaphS0 2 , 2-NaphCA or Xan.
  • preferable ones are those wherein p is an integer of 1.
  • Other preferable compounds are compounds of the formula (I) wherein Z is a single bond;
  • R 1 is selected from l-FCA, 9-FAc, Ac or Xan; r is an integer of 0; X 1 is selected from Arg or Lys; m is an integer of 1; L 1 is Cys;
  • X 2 is 3-Asp; p is an integer of 0; X 4 is ThioP; q is an integer of 1; L 2 is selected from Cys or Cys-NH 2 (only when no X 5 or X 6 groups are present) ; n is an integer of 0; and s is an integer of 0.
  • X° in structure I is absent; X 1 is Arg; X 2 is j ⁇ -Asp; X 3 is absent; X 4 is ThioP; and X 5 and X 6 are absent.
  • the cyclizing Z moiety is formed by a disulfide linkage between L 1 and L 2 cysteine residues and the overall preferred sequence would be Arg-Cys*- (S-Asp) - (ThioP) -Cys* (* denotes residue involved in cyclization of compound) .
  • R 1 is preferably l-FCA, 9-FAc, 9-FCA, Xan, For, Ac, Biotinyl, Ada, a hydrogen atom, 5-Finc, PhAc or Su.
  • residue X° is absent, residue X 1 is Lys; residue X 2 is jS-Asp; X 3 is absent; X 4 is ThioP; and X 5 and X 6 are absent.
  • sequence Lys-Cys*- ( / ⁇ -Asp) - (ThioP) -Cys* is also preferred, wherein the cyclizing Z moiety is formed by the disulfide linkage between the cysteine residues.
  • R 1 is preferably l-FCA, 9-FCA or 9-FAc or Fmoc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Arg; X 2 is 3-Asp; X 3 is absent; X 4 is ThioP; and X 5 is Ala, and n is 2 and X 6 is absent.
  • sequence Arg-Cys*- (S-Asp) - (ThioP) - Cys*-Ala-Ala is also preferred, wherein the cyclizing Z moiety is formed by a disulfide linkage between the cysteine residues.
  • R 1 is preferably l-FCA, 9-FCA, 2-NaphAc, l-NaphS0 2 , 2-NaphS0 2 , 2-NaphCA, Ac or PhAc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Lys; X 2 is ⁇ -Asp; X 3 is absent; X 4 is ThioP; X 5 is Ala and n is an integer of 2 and X 6 is absent.
  • the sequence Lys-Cys*- ( ⁇ - Asp) - (ThioP) -Cys*-Ala-Ala is also preferred, wherein the cyclization of the peptide occurs through a disulfide linkage formed between the cysteine residues.
  • R 1 is preferably 9-FCA and R 2 is PhAc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Arg; X 2 is jS-Asp; X 3 is absent; X 4 is ThioP; X 5 is Ala and X 6 is Asp.
  • R 1 is preferably Ac and R 4 is Ada or Fm.
  • sequence Orn-Cys*- (S-Asp) - (ThioP) -Cys* is also preferred, wherein the sulfhydryl linkages of the two cysteine residues cyclize the peptide.
  • R 1 is preferably l-FCA or Fmoc and the side chain terminal amino group of Orn is denoted as NHR 2
  • R 2 is preferably hydrogen or PhAc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Arg; X 2 is /S-Asp; X 3 is absent; X 4 is ThioP; X 5 is Ala and X 6 is Ser.
  • the sequence Arg-Cys*- (S-Asp) - (ThioP) -Cys*-Ala-Ser is also preferred, wherein the peptide is cyclized by a disulfide linkage of the cysteine residues.
  • R 1 is preferably l-FCA.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Lys; X 2 is / ⁇ -Asp; X 3 is absent; X 4 is TCA; and X 5 and X 6 are absent.
  • the sequence Lys-Cys*- ( / S-Asp) - (TCA) -Cys* is also preferred, wherein the peptide is cyclized by a disulfide linkage of the cysteine residues.
  • R 1 is preferably Fmoc, 9-FAc, 9-FCA or l-FCA
  • the e-amino terminal group of Lys is denoted as NHR 2 and R 2 is preferably H, 1-NaphAc, 2-NaphAc or PhAc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Lys; X 2 is /8-Asp; X 3 is absent; X 4 is Pro; X 5 and X 6 are absent.
  • the sequence Arg-Cys*- (/3-Asp) -Pro-Cys* is also preferred, wherein the peptide is cyclized by a disulfide linkage of the cysteine residue.
  • R 1 is preferably Fmoc, 9-FAc, 9-FCA or l-FCA.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Arg; X 2 is / ⁇ -Asp; X 3 is absent; X 4 is absent and X 5 and X 6 are absent.
  • the sequence Arg-Cys*- (/3-Asp) -Cys* is also preferred, wherein the cyclizing Z moiety is formed by a disulfide linkage between the cysteine residues.
  • R 1 is preferably l-FCA, 9-FCA, 9- FAc or Fmoc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Arg; X 2 is / S-Asp; X 3 is absent; X 4 is ThioP; X 5 is Abu and X 6 is absent.
  • R 1 is preferably 9-FCA, l-FCA, 9-FAc or Fmoc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Arg; X 2 is / S-Asp; X 3 is absent; X 4 is ThioP; X 5 is Ala and X 6 is absent.
  • R 1 is preferably l-FCA.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Arg; X 2 is /3-Asp; X 3 is absent; X 4 is ThioP; X 5 is Gly and n is an integer of 2 and X 6 is absent.
  • the sequence Arg-Cys*- ( ⁇ - Asp) - (ThioP) -Cys*-Gly-Gly is also preferred, wherein the cyclizing Z moiety is formed from the disulfide linkage of the cysteine residues.
  • R 1 is preferably l-FCA, 9-FCA, 9-FAc or Fmoc.
  • Another particularly preferred sequence is one wherein X° is absent; X 1 is Lys; X 2 is /8-Asp; X 3 is absent; X 4 is ThioP; X 5 is Ala and X 6 is Ser.
  • the sequence Lys-Cys*- (/8-Asp) - (ThioP) -Cys*-Ala-Ser is also preferred, wherein the cysteine residues are joined by a disulfide linkage.
  • R 1 is preferably l-FCA or 9-FCA
  • the e-amino terminal group of Lys is denoted as NHR 2 and R 2 is preferably H, 2-NaphAc, 1-NaphAc or PhAc.
  • amino acids are those from natural or non-natural sources and are those having at least one amino group and one carboxyl group in a molecule.
  • amino acids include amino acids from natural sources or antipodes thereof, D- or L-enantiomeric amino acids, and racemic mixtures of these amino acids.
  • ⁇ -Amino acids or /3-amino acids are the preferred examples and may be either one of neutral, acidic, and basic amino acids.
  • amino acids having plural amino groups or a guanidino group or an imidazolyl group such as arginine, histidine, ornithine, lysine, and the like
  • amino acids having plural carboxyl groups such as glutamic acid, aspartic acid, and the like
  • amino acids having the same number of amino groups and carboxyl groups such as alanine, isoleucine, leucine, and the like.
  • amino acids which can be used suitably, include glycine, alanine, N-methylalanine, N- ⁇ -methylarginine, isoleucine, leucine, valine, norvaline, glutamic acid, methionine, phenylalanine, proline, 8-alanine, arginine, norarginine, ornithine, serine, 0-methylserine, threonine, O-methylthreonine, asparagine, aspartic acid, jS-aspartic acid, glutamine, ⁇ -glutamine, cystine, cysteine, tyrosine, O-methyltyrosine, histidine, tryptophan, lysine, homoarginine, ⁇ -glutamic acid, sarcosine, creatine, homocysteine, norleucine, isoserine, homoserine, norvaline, ornithine, penicillamine,
  • Amino acid mimetics are derivatives of the above amino acids. Specific examples of those which can be used suitably, include ester thereof (e.g., a lower alkyl ester, aryl ester), amide thereof (e.g., unsubstituted amide, a cyclohexylamide) or derivative of proline (e.g., thioproline, 3-thiomorpholinecarboxylic acid, tetrahydro-1,3-thiazine-4-carboxylic acid, homoproline, hydroxyproline) and the like.
  • ester thereof e.g., a lower alkyl ester, aryl ester
  • amide thereof e.g., unsubstituted amide, a cyclohexylamide
  • proline e.g., thioproline, 3-thiomorpholinecarboxylic acid, tetrahydro-1,3-thiazine-4-carboxylic acid, homoproline,
  • An aryl group, an aryl moiety in the arylalkyi, aryloxycarbonyl, arylcarbonyl, arylacetyl or arylalkoxycarbonyl is preferably an aromatic hydrocarbocyclic group containing 1 to 4 rings, such as phenyl group, naphthyl group, and the like.
  • a heteroaryl group is preferably an aromatic group containing 1 or 2 rings wherein at least one of the rings contains at least 1, preferably 1 to 3, more preferably 1 or 2 heteroatoms such as N, 0 or S wherein the remainder of the ring atoms are carbon atoms, wherein each ring contains 4 to 7 ring atoms, with a total of 4 to 11 ring atoms where two fused rings are present.
  • a heteroaryl group is a 5- or 6- membered aromatic heterocyclic group such as thienyl group, furyl group, pyridyl group, pirazinyl group, pyrimidinyl group, and the like.
  • the above aryl and/or heteroaryl group may have one to three substituents and as the substituent, there may be mentioned a halogen atom, a lower alkyl group and a lower alkoxy group.
  • a lower alkyl group is preferably a C ⁇ C g alkyl group, more preferably a C-_-C 5 alkyl group
  • a lower alkanoyl group is preferably a C 1 -C 9 alkanoyl group, more preferably a C- j ⁇ -C 8 alkanoyl group
  • a lower alkoxy group is preferably a C 1 -C 8 alkoxy group, more preferably a C 1 -C 5 alkoxy group
  • aryl is preferably a C 6 -C 17 aryl group (containing 1 to 4 rings) , more preferably a C 8 -C 12 aryl group (containing 1 or 2 rings) and most preferably is an aromatic hydrocarbocyclic group containing 1 or 2 rings, such as a phenyl group, naphthyl group and the like
  • arylalkyi is preferably a C 6 -C 17 or C 6 -C 12 aryl group (containing 1 or 2
  • Aryl-O-C(O) -) more preferably a C 6 -C 13 aryloxycarbonyl (containing 1 or 2 rings)
  • arylcarbonyl is preferably an aryl group as defined above having a carbonyl group attached to one of the ring carbon atoms (e.g. Aryl-C(O)-) , more preferably C 7 - C 16 arylcarbonyl (containing 1 or 2 rings)
  • arylacetyl is preferably an aryl group as defined above having an acetyl group attached to one of the ring carbon atoms (e.g.
  • Aryl-CH 2 -C(0) -) more preferably a C 8 -C 17 arylacetyl group (containing 1 or 2 rings) and heterocyclic is preferably a heterocyclic group containing l or 2 rings containing at least 1, preferably 1 to 3, more preferably 1 or 2 heteroatoms such as N, O or S wherein the remainder of the ring atoms are carbon atoms, wherein each ring is saturated or unsaturated and contains 4 to 7 ring atoms, with a total of 4 to 11 ring atoms where two fused rings are present.
  • an effective amount of the active compound, including derivatives or salts thereof, or a pharmaceutical composition containing the same in combination with a pharmaceutical carrier or diluent, as described below, is administered via any of the usual and acceptable methods known in the art, either singly or in combination with another compound or compounds of the present invention or other pharmaceutical agents such as immunosuppressants, antihistamines, corticosteroids, and the like.
  • compositions can thus be administered orally, sublingually, topically (e.g., on the skin or in the eyes) , by inhalation, by suppository, or parenterally (e.g., intramuscularly, intravenously, subcutaneously or intradermally) , and in the form of either solid or liquid dosage including tablets, suspensions, and aerosols, as is discussed in more detail below.
  • the administration can be conducted in single unit dosage form with continuous therapy or in single dose therapy ad libitum.
  • a unit dose is defined as 1 to 3000 mg for a human patient.
  • compositions hereof can be solids, liquids or mixtures thereof; thus, the compositions can take the form of tablets, pills, capsules, powders, enterically coated or other protected formulations (such as binding on ion exchange resins or other carriers, or packaging in lipid or lipoprotein vesicles or adding additional terminal amino acids) , sustained release formulations, erodible formulations, implantable devices or components thereof, microsphere formulations, solutions (e.g., ophthalmic drops), suspensions, elixirs, aerosols, and the like.
  • protected formulations such as binding on ion exchange resins or other carriers, or packaging in lipid or lipoprotein vesicles or adding additional terminal amino acids
  • sustained release formulations such as binding on ion exchange resins or other carriers, or packaging in lipid or lipoprotein vesicles or adding additional terminal amino acids
  • erodible formulations e.g., implantable devices or components thereof
  • microsphere formulations solutions (e.g., ophthalmic drops
  • Water, saline, aqueous dextrose, and glycols are preferred liquid carriers, particularly (when isotonic) for injectable solutions.
  • the carrier can be selected from various oils including those of petroleum, animal, vegetable or synthetic origin, for example, peanut oil, soybean oil, mineral oil, sesame oil, and the like.
  • Suitable pharmaceutical excipients include starch, cellulose, talc, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate, sodium stearate, glycerol monostearate, sodium chloride, dried skim milk, glycerol, propylene glycol, water, ethanol, and the like.
  • compositions may be subjected to conventional pharmaceutical expedients such as sterilization and may contain conventional pharmaceutical additives such as preservatives, stabilizing agents, wetting or emulsifying agents, salts for adjusting osmotic pressure, buffers, and the like.
  • suitable pharmaceutical carriers and their formulations are described in Martin, "Remington's Pharmaceutical Sciences", 15th Ed.; Mack Publishing Co., Easton (1975); see, e.g., pp. 1405-1412 and pp. 1461-1487.
  • Such compositions will, in general, contain an effective amount of the active compound together with a suitable amount of carrier so as to prepare the proper dosage form for proper administration to the host.
  • the therapeutic methods of the present invention are practiced when the relief of symptoms is specifically required or perhaps imminent; in another preferred embodiment, the method hereof is effectively practiced as continuous or prophylactic treatment.
  • the particular dosage of pharmaceutical composition to be administered to the subject will depend on a variety of considerations including the nature of the disease, the severity thereof, the schedule of administration, the age and physical characteristics of the subject, and so forth. Proper dosages may be established using clinical approaches familiar to the medicinal arts. It is presently believed that dosages in the range of 0.1 to 100 mg of compound per kilogram of subject body weight will be useful, and a range of 1 to 100 mg per kg generally preferred, where administration is by injection or ingestion.
  • Topical dosages may utilize formulations containing generally as low as 0.1 mg of compound per ml of liquid carrier or excipient, with multiple daily applications being appropriate.
  • the compounds and therapeutic or pharmaceutical compositions of the invention might be useful in the study or treatment of diseases or other conditions which are mediated by the binding of integrin receptors to ligands, including conditions involving inappropriate (e.g., excessive or insufficient) binding of cells to natural or other ligands.
  • Such diseases and conditions might include inflammatory diseases such as rheumatoid arthritis, asthma, allergy conditions, adult respiratory distress syndrome, inflammatory bowel diseases (e.g., ulcerative colitis and regional enteritis) and ophthalmic inflammatory diseases; autoimmune diseases; and cardiovascular disease; prevention of occlusion following thrombolysis; neoplastic disease including metastatic conditions; as well as conditions wherein increased cell binding is desired, as in wound healing or prosthetic implantation situations, as discussed in more detail above.
  • inflammatory diseases such as rheumatoid arthritis, asthma, allergy conditions, adult respiratory distress syndrome, inflammatory bowel diseases (e.g., ulcerative colitis and regional enteritis) and ophthalmic inflammatory diseases; autoimmune diseases; and cardiovascular disease; prevention of occlusion following thrombolysis; neoplastic disease including metastatic conditions; as well as conditions wherein increased cell binding is desired, as in wound healing or prosthetic implantation situations, as discussed in more detail above.
  • the compounds of the present invention might find use in the diagnosis of diseases which result from abnormal cell adhesion.
  • diseases which result from abnormal cell adhesion For example, excessive adhesion of leukocytes to endothelial cells or to exposed extracellular matrix in blood vessels has been implicated in early stages of atherosclerosis.
  • a person demonstrating excessive binding of leukocytes to endothelial cells might be at risk for developing occluded arteries.
  • One might detect this risk factor by measuring the binding of a compound of structure I to leukocytes and endothelial cells of the patient thought to be at risk.
  • the compounds of the present invention might find use in the diagnosis of autoimmune diseases caused by antibodies which bind to cell adhesion molecules or which bind to receptors for cell adhesion molecules.
  • a disease is caused by antibodies binding to a cell adhesion molecule which is mimicked by a compound of structure I
  • a diagnostic test for the presence of such antibodies is easily performed by immunoassay of blood or serum from a patient using a compound of structure I bound to a carrier so as to capture the antibodies.
  • the bound antibody can be detected by the means typical of the art such as a labelled second antibody directed to the Fc portion of human antibodies or using labelled Fc-binding proteins from bacteria (protein A or protein G) .
  • a competitive immunoassay format can be used. In this format, the compound I is labelled and competition for binding to receptor protein attached to the substrate can be measured.
  • derivatives of the present compounds might be useful in the generation of antigens which are prepared by coupling the peptides to a carrier protein. Animals are then immunized with this complex thereby generating antibodies to the peptides. These antibodies will, in some cases, themselves be effective in inhibiting cell adhesion or modulating immune activity by acting as receptors for cell adhesion ligands, or, if anti-idiotypic, by acting to block cellular receptors.
  • the compounds of the present invention might be used to produce matrices for purifying substances which bind to the compounds of the present invention with high affinity.
  • a matrix could be produced, for example, by covalently attaching a compound of the present invention to a derivatized chromatographic support.
  • a cyclic peptide listed in Table 1 which contains a free amino group can be coupled to a cyanogen bromide activated chromatography resin, such as that available from Pharmacia, (Uppsala, Sweden, Cat. No. 52-1153-00-AK) .
  • an amino group can be introduced into the desired peptide, either by addition of a lysine residue, or by addition of another amine-containing residue.
  • carbodiimide-activated resin can be used in conjunction with cyclic peptides bearing free carboxyl functions.
  • the peptide is coupled using the protocol essentially as provided by the manufacturer.
  • the cyclic peptide-derivatized resin can then be used to purify proteins, polysaccharides or the like which may bind the cyclic peptide with high affinity.
  • Such a purification is accomplished by contacting the cyclic peptide- derivatized resin with a sample containing the compound to be affinity purified under conditions which allow formation of the specific complex, washing of the complex bound to the resin with a solution which removes unwanted substances, but leaves the complex intact, and then eluting the substance to be purified by washing the resin with a solution which disrupts the complex.
  • Boc protecting group was removed by mixing the resin with TFA:DCM (1:1) for 20 minutes. Following rinsing in order with DCM (3X) , MeOH (2X) , DCM (3X) the resin was air dried.
  • TCA When used in the peptide synthesis, TCA was obtained by the chemical synthesis procedures fully disclosed in the Published International Patent Application WO 94/15958. Those procedures are fully incorporated in their entirety into this application by reference.
  • Boc-deprotected peptides on-resin were cleaved by stirring at -5 to 0°C with a cocktail of distilled anhydrous HF (10 ml/g resin) , anisole (1 ml/g resin) and dimethyl sulfide (0.5 ml/g resin) . After one hour, the HF was removed under reduced pressure. The cleaved peptide/resin mixture was washed three times with diethyl ether and then extracted with 80% aqueous acetic acid. The combined extracts (100 ml/g or 200 ml/2g resin) were pooled and carried on to the cyclization step.
  • Cyclization The peptides were cyclized through the formation of an amide linkage, a disulfide linkage, a monosulfide linkage or a dithioether linkage, as appropriate.
  • the cyclic peptide was purified on a Waters Delta Prep 3000 system (Waters, Milford, Massachusetts) equipped with C 18 reverse phase column (15-20 mm, 5X30 cm ID) , using a linear gradient of increasing acetonitrile concentration in 1.0% triethylammonium phosphate (TEAP, pH 2.3) as the mobile phase. The appropriate fractions were pooled to give the pure peptide as a phosphate salt. The peptide salt was applied again to the column and eluted with a linear gradient of increasing amounts of acetonitrile in 1.0% aqueous HOAc to afford the desired acetate salt form.
  • TEAP triethylammonium phosphate
  • N-Boc-L- ⁇ -Asp- / ⁇ - fluoromethyl ester was attached to the hydroxymethyl resin using DCC and DMAP as a catalyst (Fehrentz, J.A. and Castro B., Synthesis. 676-678 (1983)).
  • Boc-L- ⁇ - Asp(OFm)OCH 2 -resin (1.0 g, 0.75 mmol) was used as the starting resin. Excess amino acid (2-3 fold) was used for each coupling.
  • the peptide chain was constructed on the Beckman peptide synthesizer using Boc chemistry with the stepwise addition of each amino acid following the standardized cycle similar to that presented in Table 1, with adjustments for scale. 50% TFA in DCM, 5% DIEA in DCM, and 0.5 M of DCC in DCM were used as deprotecting, neutralizing, and activating agents, respectively, for each cycle.
  • the peptide was cyclized on the resin by forming an amide linkage between the ⁇ -carboxyl group of Asp and the e-amino group of Lys using the following general procedure.
  • the final cyclic compound was removed from the resin by treatment with HF in the presence of anisole for 1 hr at 0°C. After removal of the HF, in vacuo, the residue was washed with diethyl ether and the peptide was extracted from the resin with an aqueous HOAc solution. The aqueous extract was lyophilized to yield the crude peptide.
  • the compound was purified using a Waters Delta Prep 3000 system (Waters, Milford, MA) equipped with a C 18 column, using a linear gradient of increasing acetonitrile concentration in TEAP (pH 2.2 to 2.4) as the mobile phase.
  • the collected fractions of the pure compound were pooled and applied again to the C 18 column. This time the sample was eluted with a linear gradient of increasing amounts of acetonitrile in 1.0% aqueous HOAc to convert the phosphate salt form of the peptide to the desired acetate form.
  • the pure peptide fractions were pooled, concentrated in vacuo, redissolved in water and lyophilized to give a white powder.
  • This procedure can also be used for synthesis and purification of longer cyclic peptide compounds which contain a disulfide linkage.
  • Trifluoroacetic acid was from Halocarbon Co. (New York,
  • Triethylamine was from Fisher Scientific
  • the peptide was synthesized by the solid phase method with a Beckman automated peptide synthesizer
  • Boc-Lys(Fmoc) Boc-/8-Asp( ⁇ -0-Bzl) and Boc- (ThioP) .
  • Boc-Cys(4-Me-Bzl) -0CH 2 -polystyrene resin was used for the stepwise assembly of the title peptide, using the Boc amino acid procedure in Table 1. Following capping of the N-terminus with l-FCA, the Lys N-e-Fmoc group was removed by treatment with 20% piperidine in DMF. The solution was then stirred for 20 minutes at room temperature and filtered. The resin was washed with DMF.
  • the free Lys side-chain amino group was coupled with bromoacetic acid and after HF cleavage, the peptide was cyclized by forming a mono-sulfide linkage by the general procedure, as described below.
  • the Lys side-chain deprotected peptide was coupled with bromoacetic acid through the use of DCC in DCM. The mixture was stirred for 2 hours at room temperature
  • the bromoacetyl peptide was removed from the resin by treatment with HF in the presence of anisole (1 ml/g resin) and DMS (0.5 ml/g resin) for 1 hr. at 0°C. The HF was removed under reduced pressure. The cleaved mixture was washed three times with diethyl ether and the peptide was extracted into aqueous acetic acid. This bromoacetyl peptide is unstable, and the cyclization to form the monosulfide bridge immediately followed the HF cleavage.
  • 1-Fluorenecarboxylic acid, dibromoethane, and DCC were purchased from Aldrich Chemical Company (Milwaukee, WI) . All amino acids, amino acid derivatives and analogs were purchased from BACHEM INC. (Torrance, California) . Trifluoroacetic acid was from Halocarbon Co. (New York, New York) . Triethylamine was from Fisher Scientific (Fall Lawn, New Jersey) . Other reagents were obtained from conventional sources and were of analytical grade.
  • the peptide was synthesized by the solid phase method with a Beckman automated peptide synthesizer (System 990, Beckman Instruments, Inc., Palo Alto, California) . N-Boc-S-9-Fluorenylmethyl-L-Cysteine was attached to the hydroxymethyl resin using DCC and DMAP as a catalyst. The following amino acids were used in the synthesis: Boc-Arg(Tos) , Boc- / S-Asp( ⁇ -0-Bzl) and Boc- (ThioP) . Boc-Cys(Fm) -0CH 2 -polystyrene resin was used for the stepwise assembly of the peptide following the Boc amino acid procedure in Table 1.
  • the dithioether linkage (-S- (CH 2 )n-S-) was synthesized while the peptide was bound on the resin.
  • the dithioether linkage was formed by adding dibromoethane (3 equivalents) and DIEA in DMF. The pH was adjusted to 8-9, and the solution was stirred for 2 to 4 hours at room temperature, (completion of the cyclization was monitored by HPLC and the Ellman test) . The peptide was then washed with DMF (2 x 1 min) , DCM (3 x 1 min) , MeOH (2 x 1 min) and DCM (3 x 1 min) .
  • the cyclic peptide was removed from the resin by treatment with HF in the presence of anisole for 1 hr. at 0°C. After removal of the HF, in vacuo, the residue was washed with diethyl ether and the peptide was extracted into an aqueous acetic acid solution. The aqueous extract was lyophilized to yield the crude peptide.
  • the crude peptide was purified as described for Procedure A, Section 4. TABLE 2
  • Amino acids of the peptide backbone are delineated with hyphens; synthetic amino acids unnatural amino acids, and amino acid mimetics are enclosed in parentheses; the R 1 -R 5 group which are attached to the backbone amino acid groups are enclosed in parentheses and ar
  • the following assay established the activity of the present compounds in inhibiting ⁇ -mediated cell adhesion in a representative in vitro system.
  • This assay measured the adhesive interactions of a T-cell line, Jurkat, known to express high levels of ⁇ / 8 1 , to endothelial cell monolayers in the presence of representative test compounds of the present invention.
  • the test compounds were added in increasing concentrations to T-cells and then the T-cell/compound mixture was added to IL-1 stimulated endothelial cell monolayers. The plates were incubated, washed and the percentage of attached cells was quantitated.
  • the present assay directly demonstrated the cell adhesion inhibitory activity and adhesion modulatory activity of the present compounds.
  • Human umbilical vein endothelial cells were purchased from Clonetics (San Diego, CA) at passage number 2. The cells were grown on 0.5% porcine skin gelatin pre-coated flasks (Sigma, St. Louis, MO) in EGM- UV media (Clonetics, San Diego, CA) supplemented with 10% fetal bovine serum. Cells were re-fed every 2-3 days, reaching confluence by day 4 to 6. The cells were monitored for factor VIII antigen and our results showed that at passage 12, the cells are positive for this antigen. The endothelial cells were not used following passage 7. The T-cell line Jurkat was obtained from American Type Culture Collection and cultured in RPMI media containing 10% fetal calf serum.
  • HBSS Hank's buffered saline solution
  • DMEM Dulbecco's Minimal Eagle's Media
  • HSA Human Serum Albumin
  • Confluent endothelial monolayers grown in 96-well tissue culture plates, were stimulated for 4 hours at 37°C with 0.1 ng/ml (50 U/ml) recombinant IL-1 (Amgen, Thousand Oaks, CA) . Following this incubation, the monolayers were washed twice with HBSS and 0.1 ml of DMEM-HSA solution was added.
  • Jurkat cells (5 x 10 5 cells) were combined with the appropriate concentration of peptide and 0.1 ml of the Jurkat cell-peptide mixture was added to the endothelial cell monolayers. Generally, 250, 50, 10 and 2 ⁇ M peptide concentrations were tested.
  • the IC 50 was determined by testing the peptides at 50, 10, 2 and 0.4 ⁇ M. The plates were placed on ice for 5 minutes to allow for Jurkat cell settling and the plates were incubated at 37°C for 20 minutes. Following this incubation, the monolayers were washed twice with PBS containing 1 mM calcium chloride and 1 mM magnesium chloride and the plates were read using a Millipore Cytofluor 2300 (Marlboro, MA) . Fluorescence in each well was measured as Arbitrary Fluorescence Units and percent adhesion in the absence of peptide was adjusted to 100% and the % adhesion in the presence of peptides was calculated. Monolayers were also fixed in 3% paraformaldehyde and evaluated microscopically to verify the adhesion.
  • the JY-Endothelial Cell Assay The following assay established the activity of the present compounds in inhibiting cell adhesion through ⁇ 2 integrins in a representative in vi tro system. The present assay directly demonstrated the anti- / 8 2 adhesion activity and adhesion modulatory activity of the present compounds.
  • the human B cell line, JY was cultured in RPMI media containing 10% fetal calf serum at 37°C, in a humidified C0 2 atmosphere. Endothelial cells were grown to confluency in 96-well micro-titer assay plates, as in the Jurkat-EC ' assay. Before the assay, the endothelial cells were stimulated with 50 U/ml rTNF for 18-24 hours at 37°C.
  • JY cells were loaded with the fluorescent dye indicator BCECF-AM as follows: JY cells were washed twice with HBSS and cells were resuspended in HBSS at 5X10 6 cells/ml; BCECF-AM (Molecular Probes), stock concentration ⁇ mg/ml in DMSO, was added to the JY cells to a final concentration of 2 ⁇ g/ml; cells were incubated in the dark at 37°C for 30-45 minutes; washed twice with HBSS and used in the assay.
  • the compounds presented in this invention were typically dissolved in 2.5 mg HSA/ml DME at four times the assay concentration and Ph adjusted with 7.5% Na bicarbonate, as needed.
  • TNF-stimulated HUVEC were washed twice with HBSS (at room temp) .
  • 50 ⁇ l of cold HSA-DME was added to each well and the plate transferred to ice. While on ice, the following components were added to the wells in the indicated order:
  • test compound 2 50 ⁇ l BCECF-labelled JY cells
  • the above-described adhesion assay yielded the following results for the inhibition of Jurkat cell adhesion to IL-1 stimulated endothelial cells.
  • an objective of the present invention is to provide compounds having extraordinarily high potencies in modulating cell adhesion to VCAM and ICAM through ⁇ and ⁇ 2 integrins, respectively, including but not limited to inhibition of T-cell adhesion to endothelial cells.
  • the exact receptors involved in this interaction and the specific receptors targeted by the test compounds include, but are not limited to, a ⁇ i 0* 4/ 8 7 ,
  • the amino acid without an indication of enantiomeric structure means L 25 enantiomer.

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Abstract

L'invention concerne des composés peptidiques cycliques qui modulent l'adhérence cellulaire induite par intégrine. Ces composés peuvent inhiber l'adhérence cellulaire induite à la fois par β1 et β2. L'invention concerne en outre des applications thérapeutiques de ces composés dans le traitement de pathologies apparentées à l'adhérence cellulaire.
PCT/US1996/010186 1995-06-07 1996-06-06 INHIBITEURS PEPTIDIQUES CYCLIQUES D'ADHERENCE CELLULAIRE INDUITE PAR INTEGRINE β1 ET β¿2? Ceased WO1996040781A1 (fr)

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AU63321/96A AU6332196A (en) 1995-06-07 1996-06-06 Cyclic peptide inhibitors of beta1 and beta2 integrin-mediated adhesion
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US6020347A (en) * 1997-11-18 2000-02-01 Merck & Co., Inc. 4-substituted-4-piperidine carboxamide derivatives
US6031072A (en) * 1996-07-12 2000-02-29 Mcgill University Compounds and methods for modulating cell adhesion
US6069163A (en) * 1997-10-21 2000-05-30 Merck & Co., Inc. Azapeptide acids as cell adhesion inhibitors
US6090841A (en) * 1997-11-21 2000-07-18 Merck & Co., Inc. Substituted pyrrole derivatives as cell adhesion inhibitors
US6169071B1 (en) 1996-07-12 2001-01-02 Mcgill University Compounds and methods for modulating cell adhesion
US6191171B1 (en) 1997-11-20 2001-02-20 Merck & Co., Inc. Para-aminomethylaryl carboxamide derivatives
US6203788B1 (en) 1997-09-29 2001-03-20 Adherex Inc. Compounds and methods for regulating cell adhesion
US6207639B1 (en) 1996-07-12 2001-03-27 Mcgill University Compounds and methods for modulating neurite outgrowth
US6221888B1 (en) 1997-05-29 2001-04-24 Merck & Co., Inc. Sulfonamides as cell adhesion inhibitors
US6277824B1 (en) 1998-07-10 2001-08-21 Adherex Technologies Compounds and methods for modulating adhesion molecule function
US6291511B1 (en) 1997-05-29 2001-09-18 Merck & Co., Inc. Biarylalkanoic acids as cell adhesion inhibitors
US6333307B1 (en) 1996-07-12 2001-12-25 Mcgill University Compounds and method for modulating neurite outgrowth
US6346512B1 (en) 1996-07-12 2002-02-12 Mcgill University Compounds and methods for modulating cell adhesion
US6417325B1 (en) 1996-07-12 2002-07-09 Mcgill University Compounds and methods for cancer therapy
EP1015479A4 (fr) * 1997-04-11 2002-07-24 Lilly Co Eli Echantillotheques combinatoires de macrocycles peptidomimetiques et procedes correspondants
US6465427B1 (en) 1996-07-12 2002-10-15 Mcgill University Compounds and methods for modulating cell adhesion
US6511961B1 (en) 1997-11-13 2003-01-28 Toray Industries, Inc. Cyclic peptides and medicinal use thereof
RU2203902C2 (ru) * 1997-07-11 2003-05-10 Байотай Терапис Корп. Связывающийся с интегрином пептид и его применение
US6562786B1 (en) 1996-07-12 2003-05-13 Mcgill University Compounds and methods for modulating apoptosis
US6610821B1 (en) 1996-07-12 2003-08-26 Mcgill University Compounds and methods for modulating endothelial cell adhesion
US6645939B1 (en) 1997-11-24 2003-11-11 Merck & Co., Inc. Substituted β-alanine derivatives as cell adhesion inhibitors
US6667331B2 (en) 1999-12-28 2003-12-23 Pfizer Inc Non-peptidyl inhibitors of VLA-4 dependent cell binding useful in treating inflammatory, autoimmune, and respiratory diseases
WO2004041279A1 (fr) 2002-10-30 2004-05-21 Merck & Co., Inc. Modulateurs gamma-aminoamides de l'activite de recepteur de chimiokine
US6903075B1 (en) 1997-05-29 2005-06-07 Merck & Co., Inc. Heterocyclic amide compounds as cell adhesion inhibitors
US7122623B2 (en) 1996-07-12 2006-10-17 Adherex Technologies, Inc. Compounds and methods for modulating cell adhesion
EP2045241A1 (fr) 2002-01-24 2009-04-08 Merck Frosst Canada Ltd. Cycloalkanoindoles substitues par un fluor et leur utilisation comme antagonistes du recepteur de prostaglandine D2
WO2009126920A2 (fr) 2008-04-11 2009-10-15 Merrimack Pharmaceuticals, Inc. Lieurs d'albumine de sérum humain, et ses conjugués
EP2140881A1 (fr) 1999-12-16 2010-01-06 Biogen Idec MA Inc. Procédés de traitement de maladie hémorragique ou ischémique du système nerveux central, utilisant des antagonistes d'intégrine anti alpha 4
WO2010031183A1 (fr) 2008-09-22 2010-03-25 Merck Frosst Canada Ltd. Dérivés d'indole comme antagonistes du récepteur crth2
WO2010090764A1 (fr) 2009-02-09 2010-08-12 Supergen, Inc. Inhibiteurs pyrrolopyrimidinyle de l'axi kinase
WO2012051036A1 (fr) 2010-10-11 2012-04-19 Merck Sharp & Dohme Corp. Composés de type quinazolinone convenant comme antagonistes de crth2
WO2012087861A1 (fr) 2010-12-23 2012-06-28 Merck Sharp & Dohme Corp. Quinoxalines et aza-quinoxalines comme modulateurs du récepteur crth2
EP2492267A1 (fr) 2009-02-24 2012-08-29 Merck Sharp & Dohme Corp. Dérivés d'indole en tant qu'antagonistes du récepteur CRTH2 en combination avec un second principe actif
EP2510941A2 (fr) 2007-02-20 2012-10-17 Merrimack Pharmaceuticals, Inc. Procédés de traitement de la sclérose en plaques par administration d'une alpha-foetoprotéine combinée à un antagoniste de l'intégrine
WO2012151137A1 (fr) 2011-05-04 2012-11-08 Merck Sharp & Dohme Corp. Inhibiteurs de tyrosine kinase splénique contenant une aminopyridine
WO2012174176A1 (fr) 2011-06-17 2012-12-20 Merck Sharp & Dohme Corp. Tétrahydroquinolines à fusion cycloalkyle en tant que modulateurs de l'activité du récepteur crth2
WO2013052393A1 (fr) 2011-10-05 2013-04-11 Merck Sharp & Dohme Corp. Inhibiteurs de tyrosine kinase de la rate (syk) contenant un 3-pyridyl carboxamide
WO2018140510A1 (fr) 2017-01-25 2018-08-02 Biogen Ma Inc. Composition et méthodes de traitement d'un accident vasculaire cérébral et d'autres troubles du snc
WO2020143793A1 (fr) 2019-01-10 2020-07-16 石药集团中奇制药技术（石家庄）有限公司 Sels de composé hétérocyclique et utilisation associée

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WO1992000995A1 (fr) * 1990-07-09 1992-01-23 Tanabe Seiyaku Co. Ltd. Composes de modulation de l'adherence cellulaire cyclique
WO1994015958A2 (fr) * 1993-01-08 1994-07-21 Tanabe Seiyaku Co., Ltd. Inhibiteurs peptidiques de l'adhesion cellulaire

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Publication number Priority date Publication date Assignee Title
WO1992000995A1 (fr) * 1990-07-09 1992-01-23 Tanabe Seiyaku Co. Ltd. Composes de modulation de l'adherence cellulaire cyclique
WO1994015958A2 (fr) * 1993-01-08 1994-07-21 Tanabe Seiyaku Co., Ltd. Inhibiteurs peptidiques de l'adhesion cellulaire

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Publication number Priority date Publication date Assignee Title
US6417325B1 (en) 1996-07-12 2002-07-09 Mcgill University Compounds and methods for cancer therapy
US7122623B2 (en) 1996-07-12 2006-10-17 Adherex Technologies, Inc. Compounds and methods for modulating cell adhesion
US6780845B2 (en) 1996-07-12 2004-08-24 Mcgill University Compounds and methods for cancer therapy
US6914044B2 (en) 1996-07-12 2005-07-05 Mcgill University Compounds and methods for modulating cell adhesion
US6967238B2 (en) 1996-07-12 2005-11-22 Mcgill University Compounds and methods for modulating cell adhesion
US6346512B1 (en) 1996-07-12 2002-02-12 Mcgill University Compounds and methods for modulating cell adhesion
US6610821B1 (en) 1996-07-12 2003-08-26 Mcgill University Compounds and methods for modulating endothelial cell adhesion
US6207639B1 (en) 1996-07-12 2001-03-27 Mcgill University Compounds and methods for modulating neurite outgrowth
US6562786B1 (en) 1996-07-12 2003-05-13 Mcgill University Compounds and methods for modulating apoptosis
US7138369B2 (en) 1996-07-12 2006-11-21 Mcgill University Compounds and methods for modulating apoptosis
US6465427B1 (en) 1996-07-12 2002-10-15 Mcgill University Compounds and methods for modulating cell adhesion
US6326352B1 (en) 1996-07-12 2001-12-04 Mcgill University Compounds and methods for modulating cell adhesion
US6031072A (en) * 1996-07-12 2000-02-29 Mcgill University Compounds and methods for modulating cell adhesion
US6333307B1 (en) 1996-07-12 2001-12-25 Mcgill University Compounds and method for modulating neurite outgrowth
US6169071B1 (en) 1996-07-12 2001-01-02 Mcgill University Compounds and methods for modulating cell adhesion
EP1015479A4 (fr) * 1997-04-11 2002-07-24 Lilly Co Eli Echantillotheques combinatoires de macrocycles peptidomimetiques et procedes correspondants
US6291511B1 (en) 1997-05-29 2001-09-18 Merck & Co., Inc. Biarylalkanoic acids as cell adhesion inhibitors
US6903075B1 (en) 1997-05-29 2005-06-07 Merck & Co., Inc. Heterocyclic amide compounds as cell adhesion inhibitors
US6221888B1 (en) 1997-05-29 2001-04-24 Merck & Co., Inc. Sulfonamides as cell adhesion inhibitors
RU2203902C2 (ru) * 1997-07-11 2003-05-10 Байотай Терапис Корп. Связывающийся с интегрином пептид и его применение
US7326686B2 (en) 1997-09-29 2008-02-05 Adherex Inc. Compounds and methods for regulating cell adhesion
US6203788B1 (en) 1997-09-29 2001-03-20 Adherex Inc. Compounds and methods for regulating cell adhesion
US6069163A (en) * 1997-10-21 2000-05-30 Merck & Co., Inc. Azapeptide acids as cell adhesion inhibitors
US6511961B1 (en) 1997-11-13 2003-01-28 Toray Industries, Inc. Cyclic peptides and medicinal use thereof
US6020347A (en) * 1997-11-18 2000-02-01 Merck & Co., Inc. 4-substituted-4-piperidine carboxamide derivatives
US6191171B1 (en) 1997-11-20 2001-02-20 Merck & Co., Inc. Para-aminomethylaryl carboxamide derivatives
US6090841A (en) * 1997-11-21 2000-07-18 Merck & Co., Inc. Substituted pyrrole derivatives as cell adhesion inhibitors
US6645939B1 (en) 1997-11-24 2003-11-11 Merck & Co., Inc. Substituted β-alanine derivatives as cell adhesion inhibitors
US6806255B2 (en) 1998-07-10 2004-10-19 Adherex Technologies, Inc. Compounds and methods for modulating adhesion molecule function
US6472368B1 (en) 1998-07-10 2002-10-29 Adherex Technologies, Inc. Compounds and methods for modulating adhesion molecule function
US6277824B1 (en) 1998-07-10 2001-08-21 Adherex Technologies Compounds and methods for modulating adhesion molecule function
EP2140881A1 (fr) 1999-12-16 2010-01-06 Biogen Idec MA Inc. Procédés de traitement de maladie hémorragique ou ischémique du système nerveux central, utilisant des antagonistes d'intégrine anti alpha 4
EP2332578A1 (fr) 1999-12-16 2011-06-15 Biogen Idec MA Inc. Procédés de traitement de maladie hémorragique ou ischémique du système nerveux central, utilisant des antagonistes de l'intégrine alpha 4
US6668527B2 (en) 1999-12-28 2003-12-30 Pfizer Inc. Non-peptidyl inhibitors of VLA-4 dependent cell binding useful in treating inflammatory, autoimmune, and respiratory diseases
US6667331B2 (en) 1999-12-28 2003-12-23 Pfizer Inc Non-peptidyl inhibitors of VLA-4 dependent cell binding useful in treating inflammatory, autoimmune, and respiratory diseases
US6903128B2 (en) 1999-12-28 2005-06-07 Pfizer Inc Non-peptidyl inhibitors of VLA-4 dependent cell binding useful in treating inflammatory, autoimmune, and respiratory diseases
EP2045241A1 (fr) 2002-01-24 2009-04-08 Merck Frosst Canada Ltd. Cycloalkanoindoles substitues par un fluor et leur utilisation comme antagonistes du recepteur de prostaglandine D2
EP2295409A1 (fr) 2002-01-24 2011-03-16 Merck Frosst Canada Limited Cycloalkanoindoles substitués par un fluor et leur utilisation comme antagonistes du récepteur de prostaglandine D2
WO2004041279A1 (fr) 2002-10-30 2004-05-21 Merck & Co., Inc. Modulateurs gamma-aminoamides de l'activite de recepteur de chimiokine
EP2510941A2 (fr) 2007-02-20 2012-10-17 Merrimack Pharmaceuticals, Inc. Procédés de traitement de la sclérose en plaques par administration d'une alpha-foetoprotéine combinée à un antagoniste de l'intégrine
WO2009126920A2 (fr) 2008-04-11 2009-10-15 Merrimack Pharmaceuticals, Inc. Lieurs d'albumine de sérum humain, et ses conjugués
EP2860260A1 (fr) 2008-04-11 2015-04-15 Merrimack Pharmaceuticals, Inc. Lieurs d'albumine de sérum humain et de leurs conjugués
WO2010031183A1 (fr) 2008-09-22 2010-03-25 Merck Frosst Canada Ltd. Dérivés d'indole comme antagonistes du récepteur crth2
WO2010090764A1 (fr) 2009-02-09 2010-08-12 Supergen, Inc. Inhibiteurs pyrrolopyrimidinyle de l'axi kinase
EP2492267A1 (fr) 2009-02-24 2012-08-29 Merck Sharp & Dohme Corp. Dérivés d'indole en tant qu'antagonistes du récepteur CRTH2 en combination avec un second principe actif
WO2012051036A1 (fr) 2010-10-11 2012-04-19 Merck Sharp & Dohme Corp. Composés de type quinazolinone convenant comme antagonistes de crth2
WO2012087861A1 (fr) 2010-12-23 2012-06-28 Merck Sharp & Dohme Corp. Quinoxalines et aza-quinoxalines comme modulateurs du récepteur crth2
WO2012151137A1 (fr) 2011-05-04 2012-11-08 Merck Sharp & Dohme Corp. Inhibiteurs de tyrosine kinase splénique contenant une aminopyridine
WO2012174176A1 (fr) 2011-06-17 2012-12-20 Merck Sharp & Dohme Corp. Tétrahydroquinolines à fusion cycloalkyle en tant que modulateurs de l'activité du récepteur crth2
WO2013052393A1 (fr) 2011-10-05 2013-04-11 Merck Sharp & Dohme Corp. Inhibiteurs de tyrosine kinase de la rate (syk) contenant un 3-pyridyl carboxamide
WO2018140510A1 (fr) 2017-01-25 2018-08-02 Biogen Ma Inc. Composition et méthodes de traitement d'un accident vasculaire cérébral et d'autres troubles du snc
WO2020143793A1 (fr) 2019-01-10 2020-07-16 石药集团中奇制药技术（石家庄）有限公司 Sels de composé hétérocyclique et utilisation associée

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