WO2001042192A2 - Vla-4 integrin antagonists - Google Patents
Vla-4 integrin antagonists Download PDFInfo
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- WO2001042192A2 WO2001042192A2 PCT/EP2000/012226 EP0012226W WO0142192A2 WO 2001042192 A2 WO2001042192 A2 WO 2001042192A2 EP 0012226 W EP0012226 W EP 0012226W WO 0142192 A2 WO0142192 A2 WO 0142192A2
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- 0 C*N(CC(N*C(O)=O)=O)C(Cc(cc1)ccc1NC(Nc1c(*)cccc1)=O)=O Chemical compound C*N(CC(N*C(O)=O)=O)C(Cc(cc1)ccc1NC(Nc1c(*)cccc1)=O)=O 0.000 description 2
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
- C07D295/16—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms
- C07D295/18—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms acylated on ring nitrogen atoms by radicals derived from carboxylic acids, or sulfur or nitrogen analogues thereof
- C07D295/182—Radicals derived from carboxylic acids
- C07D295/185—Radicals derived from carboxylic acids from aliphatic carboxylic acids
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P27/00—Drugs for disorders of the senses
- A61P27/02—Ophthalmic agents
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C275/00—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups
- C07C275/28—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton
- C07C275/42—Derivatives of urea, i.e. compounds containing any of the groups, the nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of urea groups bound to carbon atoms of six-membered aromatic rings of a carbon skeleton being further substituted by carboxyl groups
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C279/00—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups
- C07C279/28—Derivatives of guanidine, i.e. compounds containing the group, the singly-bound nitrogen atoms not being part of nitro or nitroso groups having nitrogen atoms of guanidine groups bound to cyano groups, e.g. cyanoguanidines, dicyandiamides
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D295/00—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms
- C07D295/04—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms
- C07D295/12—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms
- C07D295/125—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings
- C07D295/13—Heterocyclic compounds containing polymethylene-imine rings with at least five ring members, 3-azabicyclo [3.2.2] nonane, piperazine, morpholine or thiomorpholine rings, having only hydrogen atoms directly attached to the ring carbon atoms with substituted hydrocarbon radicals attached to ring nitrogen atoms substituted by singly or doubly bound nitrogen atoms with the ring nitrogen atoms and the substituent nitrogen atoms attached to the same carbon chain, which is not interrupted by carbocyclic rings to an acyclic saturated chain
Definitions
- VLA-4 INTEGRIN ANTAGONISTS
- Cell adhesion (i.e., a process by which cells associate with each other, migrate towards a specific target, or localize within the extracellular matrix) underlies many biological phenomena. Cell adhesion causes adhesion of hemoatopoietic to endothelial cells and the subsequent migration of those hemopoietic cells out of blood vessels and to the site of injury, thus playing a role in mammalian pathologies such as inflammation and immune reactions.
- integrins are the key mediators in adhesive interactions between hematopoietic and other cells.
- Integrins are non- covalent heterodimeric complexes consisting of two subunits, ⁇ and ⁇ . Depending on the type of its and ⁇ subunit components, each integrin molecule is categorized into its own subfamily. There are at least 12 different ⁇ subunits ( ⁇ 1- ⁇ 6, ⁇ -L, ⁇ -M, ⁇ -X, ⁇ - IIB, ⁇ -V, and ⁇ -E) and at least 9 different ⁇ subunits ( ⁇ 1- ⁇ 9).
- the integrin VLA-4 (very late antigen-4), also known as ⁇ 4 ⁇ 1 integrin or CD49d/CD29, is a leukocyte cell surface receptor that participates in a variety of cell-cell and cell- matrix adhesions. It is a receptor for both the cytokine-inducible endothelial cell surface protein, vascular cell adhesion molecule-1 (VCAM-1 ), and the extracellular matrix protein fibronectin (FN).
- VCAM-1 vascular cell adhesion molecule-1
- FN extracellular matrix protein fibronectin
- Anti-VLA-4 monoclonal antibodies mAb's inhibit VLA-4- dependent adhesive interactions both in vitro and in vivo. The inhibition of VLA-4- dependent cell adhesion may prevent or inhibit several inflammatory and autoimmune pathologies.
- VLA-4 Antagonists are disclosed e.g. in International Application WO 96/22966.
- the invention relates to the compounds as defined herein which are potent VLA-4 antagonists and methods for preparation thereof, pharmaceutical compositions comprising said compounds, and methods of inhibiting VLA-4 and of treating, preventing or suppressing conditions in mammals which are responsive to VLA-4 inhibition using said compounds or pharmaceutical compositions comprising said compounds.
- the compounds of the invention are useful to inhibit, prevent and suppress VLA-4 mediated cell adhesions. They are thus useful for the treatment of VLA-4 mediated conditions, particularly inflammation, autoimmune diseases, and immune reactions to e.g. organ transplantation. Conditions in which VLA-4 is implicated include rheumatoid arthritis, respiratory diseases such as asthma, multiple sclerosis, and complications of organ transplantation, e.g. of heart, lung, pancreas (islet) transplantation.
- the invention relates to the novel compounds of formula I
- Ar is carbocyclic or heterocyclic aryl, or biaryl
- Q is O, S or N-C ⁇ N
- X is arylene
- V is NH, O, NHOH, CH 2 or a direct bond
- W is NH, O, NHOH, CH 2 or a direct bond
- Alk is C 2 -C 7 -alkylene or C 2 -C 7 -alkylene interrupted by O, S, SO, SO 2 or NR 3 ;
- Y is amino, acylamino, mono- or di- (lower alkyl, aryl or aralkyl)-amino, pyrrolidino, perhydroazepino or a group of the formula — M, M 2
- M is N; and M 2 is CH 2 , O, NR 3 , S, SO or SO 2;
- R L R 2 and R 3 are independently hydrogen, lower alkyl, lower alkenyl, cycloalkyl, aryl, cycloalkyl-lower alkyl, aryl-lower alkyl or aryl-lower alkenyl;
- Z is lower alkylene or lower alkylene substituted by one or more of lower alkyl, lower alkenyl, cycloalkyl, aryl, cycloalkyl-lower alkyl, aryl-lower alkyl or aryl-lower alkenyl; or Z is lower alkylene interrupted by O, S, SO, SO 2 or NR 3 ; pharmaceutically acceptable esters and amides thereof; and pharmaceutically acceptable salts thereof.
- Compounds of the invention may possess one or more asymmetric carbon atoms and can exist as diastereomers, racemates and the enantiomers thereof, all of which are within the purview of the invention.
- V and W are NH or NHOH; or wherein V is CH 2 and W is NH; or wherein V is a direct bond and W is NH; or wherein V is NH and W is CH 2 ; also to the above compounds wherein Q is O, S or N-C ⁇ N.
- V and W are NH; Q is O; X is phenylene; Ar is carbocyclic or heterocyclic aryl; Alk is C 2 -C 4 -alkylene; Ri, R 2 and R 3 are hydrogen or lower alkyl; Y is a group of the formula
- M ⁇ is N and M 2 is CH 2 , O or S;
- Z is C 2 -C 5 - straight chain alkylene optionally substituted by lower alkyl, lower alkenyl, carbocyclic aryl or heterocyclic aryl; or Z is C 2 - C 5 - straight chain alkylene interrupted by O, S, SO or SO 2 ; pharmaceutically acceptable esters and amides thereof; and pharmaceutically acceptable salts thereof.
- Ar is monocarbocyclic aryl;
- Alk is C 2 -C 4 -alkylene;
- R 4 is lower alkyl, lower alkenyl, or monocarbocyclic aryl; m is 1 or 2; pharmaceutically acceptable esters thereof; and pharmaceutically acceptable salts thereof.
- R 4 is phenyl or phenyl substituted by one to three of Crd-alkoxy, chloro, fluoro or C C 4 -alkyl; and R 5 is C ⁇ C ⁇ alkoxy, chloro, fluoro, or C r C 4 -alkyl; pharmaceutically acceptable esters thereof; and pharmaceutically acceptable salts thereof.
- the general definitions used herein have the following meaning within the scope of the present invention.
- Aryl represents carbocyclic or heterocyclic aryl, either monocyclic or bicyclic.
- Arylene is an aryl linking group in which aryl is heterocyclic or carbocyclic aryl, preferably monocyclic carbocyclic aryl.
- a carbocyclic aryl linking group (as for X in formula I) is for instance optionally substituted phenylene to which the two adjacent groups are attached either ortho, meta or para, preferably para, to each other.
- Monocyclic carbocyclic aryl represents optionally substituted phenyl, being preferably phenyl or phenyl substituted by one to three substituents, such being advantageously lower alkyl, hydroxy, lower alkoxy, acyloxy, halogen, cyano, trifluoromethyl, carbocyclic aryloxy or carbocyclic aryl-lower alkoxy.
- Bicyclic carbocyclic aryl represents 1 - or 2-naphthyl or 1 - or 2-naphthyl substituted by, e.g., lower alkyl, lower alkoxy or halogen.
- Monocyclic heterocyclic aryl represents optionally substituted thienyl, furanyl, pyridyl, pyrrolyl, thiazolyl, pyrazinyl, pyridazinyl or pyrazolyl, preferably optionally substituted thiazolyl, thienyl, furanyl or pyridyl.
- Optionally substituted furanyl represents 2- or 3-furanyl preferably substituted by lower alkyl.
- Optionally substituted pyridyl represents 2-, 3- or 4-pyridyl or 2-, 3- or 4-pyridyl preferably substituted by lower alkyl, halogen or cyano.
- Optionally substituted thienyl represents 2- or 3-thienyl or 2- or 3-thienyl preferably substituted by lower alkyl.
- Optionally substituted thiazolyl represents, e.g., 4-thiazolyl, or 4-thiazolyl substituted by lower alkyl.
- Bicyclic heterocyclic aryl represents e.g. quinolinyl, isoquinolinyl, indolyl or benzothiazolyl optionally substituted by hydroxy, lower alkyl, lower alkoxy or halogen.
- Aryl as in aryl-lower alkyl is preferably phenyl or phenyl substituted by one or two of lower alkyl, lower alkoxy, hydroxy, acyloxy, halogen, trifluoromethyl or cyano; also, optionally substituted naphthyl.
- Aryl-lower alkyl is advantageously benzyl or 1 - or 2-phenethyl optionally substituted on phenyl by one or two of lower alkyl, lower alkoxy, hydroxy, lower alkanoyloxy, halogen, cyano or trifluoromethyl.
- Biaryl represents phenyl substituted by carbocyclic aryl or heterocyclic aryl as defined herein, ortho, meta or para to the point of attachment of the phenyl ring, advantageously para, such as 4-biphenyl.
- a lower alkyl group preferably contains 1-4 carbon atoms and represents for example, ethyl, propyl, butyl or advantageously methyl.
- a lower alkylene group preferably contains 1 -4 carbon atoms, and represents, for example, ethylene, propylene and the like.
- a lower alkenyl group preferably contains 2-4 carbon atoms, and represents, for example, allyl.
- Cycloalkyl represents preferably cyclopentyl, cyclohexyl or cycloheptyl.
- a lower alkoxy group preferably contains 1 -4 carbon atoms and represents for example, methoxy, propoxy, isoproproxy or advantageously ethoxy.
- Halogen preferably represents fluoro or chloro, but may also be bromo or iodo.
- Acyl is derived from a carboxylic acid or carbonic acid and represents preferably optionally substituted lower alkanoyl, aroyl , lower alkoxycarbonyl or aryl-lower alkoxycarbonyl, advantageously aroyl.
- Lower alkanoyl is preferably acetyl, propionyl, butyryl, or pivaloyl.
- Optionally substituted lower alkanoyl for example represents lower alkanoyl or lower alkanoyl substituted e.g. by lower alkoxycarbonyl, lower alkanoyloxy, lower alkanoylthio, lower alkoxy, or by lower alkylthio.
- Aroyl is preferably monocyclic carbocyclic or monocyclic heterocyclic aroyl.
- Monocyclic carbocyclic aroyl is preferably benzoyl or benzoyl substituted by lower alkyl, lower alkoxy, halogen or trifluoromethyl.
- Monocyclic heterocyclic aroyl is preferably pyridylcarbonyl or thienylcarbonyl.
- Acyloxy is preferably optionally substituted lower alkanoyloxy, lower alkoxycarbonyloxy, monocyclic carbocyclic aroyloxy or monocyclic heterocyclic aroyloxy.
- Aryl-lower alkoxycarbonyl is preferably monocyclic carbocyclic-lower alkoxycarbonyl, advantageously benzyloxycarbonyl.
- esters are preferably prodrug ester derivatives, such being convertible by solvolysis or under physiological conditions to the free carboxyclic acids of formula I.
- Pharmaceutically acceptable esters are preferably prodrug esters, e.g. lower alkyl esters, cycloalkyl esters, lower alkenyl esters, benzyl esters, mono or disubstituted lower alkyl esters, e.g.
- amides are e.g. primary, secondary and tertiary amides, i.e. the unsubstituted, the N-mono-lower alkyl or N,N-di-lower alkylamides or amides of cyclic amines, e.g. of piperidine, pyrrolidine, morpholine or optionally substituted piperazine.
- Pharmaceutically acceptable salts of the acids of the invention are salts derived from pharmaceutically acceptable bases, e.g. alkali metal salts (e.g. sodium, potassium salts), alkaline earth metal salts (e.g. magnesium, calcium salts) amine salts (e.g. ethanolamine, diethanolamine, lysine and tromethamine salts) and the like conventionally used in the art.
- alkali metal salts e.g. sodium, potassium salts
- alkaline earth metal salts e.g. magnesium, calcium salts
- amine salts e.g. ethanolamine, diethanolamine, lysine and tromethamine salts
- salts may also be prepared from pharmaceutically acceptable non-toxic acids, such as mineral acids, e.g. hydrochloric, sulfuric or phosphoric acid, or organic acids, for example aliphatic or aromatic carboxylic or sulfonic acids, e.g. succinic, lactic, malic, citric, maleic, fumaric, methanesulfonic, acetic acid and the like.
- mineral acids e.g. hydrochloric, sulfuric or phosphoric acid
- organic acids for example aliphatic or aromatic carboxylic or sulfonic acids, e.g. succinic, lactic, malic, citric, maleic, fumaric, methanesulfonic, acetic acid and the like.
- the compounds of the invention are particularly useful in mammals as VLA-4 antagonists, for inhibiting VLA-4 associated cell adhesions and for treating, ameliorating or preventing conditions in mammals in which VLA-4 is implicated, e.g. immune, autoimmune disorders and inflammatory conditions.
- VLA-4 e.g. immune, autoimmune disorders and inflammatory conditions.
- respiratory disorders e.g. asthma
- arthritis e.g. rheumatoid arthritis
- psoriasis transplantation rejection
- multiple sclerosis type I diabetes
- inflammatory bowel disease e.g. rheumatoid arthritis
- psoriasis transplantation rejection
- multiple sclerosis type I diabetes
- inflammatory bowel disease e.g. sickle cell anemia, Crohn's disease, uveitis, systemic lupus erythematosus, myasthenia gravis, gout, and the like.
- the compounds of the invention are useful as agents for the symptomatic or prophylactic treatment of inflammatory airways diseases.
- Such diseases include asthma of whatever type or genesis including both intrinsic and, especially, extrinsic asthma. They are useful for the treatment of allergic asthma, whether atopic (i.e. IgE-mediated) or non-atopic, as well as bronchitic asthma, exercise- induced asthma, occupational asthma, asthma induced following bacterial infection and other non-allergic asthmas.
- Treatment of asthma is also to be understood as embracing treatment of patients of less than 4 or 5 years of age exhibiting wheezing symptoms, particularly at night and diagnosed or diagnosable as "whez infants".
- Prophylactic efficacy in the treatment of asthma may be manifested by reduced frequency or reduced severity of symptomatic attack. It may be further evidenced by reduced requirement for symptomatic therapy, i.e. therapy for, or intended to restrict or abort, symptomatic attack when it occurs, for example for anti-inflammatory therapy using a corticosteroid.
- symptomatic therapy i.e. therapy for, or intended to restrict or abort, symptomatic attack when it occurs, for example for anti-inflammatory therapy using a corticosteroid.
- pneumoconiosis an inflammatory, commonly occupational, disease of the lungs occasioned by repeated inhalation of dusts
- pneumoconiosis an inflammatory, commonly occupational, disease of the lungs occasioned by repeated inhalation of dusts
- aluminosis asbestosis, chalicosis, siderosis, silicosis, tabacosis and byssinosis.
- inflammatory airways diseases which may be treated with compounds of the invention include adult respiratory distress syndrome (ARDS), chronic obstructive pulmonary disease (COPD) in the exacerbation phase thereof and exacerbation of airways hyperactivity consequent to other drug therapy, e.g. aspirin or ⁇ -agonist bronchodilator therapy.
- ARDS adult respiratory distress syndrome
- COPD chronic obstructive pulmonary disease
- compounds of the invention are also useful for the treatment of related disorders of the airways, e.g. eosinophilia, hypereosinophilia, eosinophilic pneumonia, parasitic infestation (including tropical eosinophilia), bronchopulmonary aspergillosis, polyarteritis nodosa, eosinophilic granuloma and eosinophil-related disorders affecting the airways caused by drug- reaction.
- related disorders of the airways e.g. eosinophilia, hypereosinophilia, eosinophilic pneumonia, parasitic infestation (including tropical eosinophilia), bronchopulmonary aspergillosis, polyarteritis nodosa, eosinophilic granuloma and eosinophil-related disorders affecting the airways caused by drug- reaction.
- Compounds of the invention may also be used in the treatment of allergic inflammatory diseases such as allergic rhinitis.
- the compounds of the invention are particularly useful for inhibiting transplant rejection by transplant recipients (e.g., in heart, lung, combined heart-lung, liver, heart, kidney, pancreas, skin or corneal transplants), including both allo- and xeno-graft rejection.
- transplant recipients e.g., in heart, lung, combined heart-lung, liver, heart, kidney, pancreas, skin or corneal transplants
- the compounds of the invention are also indicated for the prevention of graft-versus- host disease, such as following bone marrow transplantation.
- the compounds of the invention may be used alone or in combination with known immunosuppressive agents.
- immunosuppressive agents include cyclosporine, tacrolimus, mycophenolic acid (mycophenolate mofetil), brequinar (brequinar sodium)), rapamycin and the like.
- the dose of these immunosuppressive agents required to achieve an immunosuppressive effect when used in combination may then be reduced, thus reducing the incidence of undesirable side effects associated with the particular known immunosuppressive agent, e.g., nephrotoxicity in the case of cyclosporine and tacrolimus.
- the above-cited properties are demonstrable in vitro and in vivo tests using advantageously mammals, e.g. rats, mice, dogs, monkeys, and isolated cells thereof.
- Said compounds can be applied in vitro in the form of solutions, e.g., preferably aqueous solutions and in vivo either enterally or parenterally, advantageously orally and intravenously.
- the dosage in vitro may range between about 10 "5 and 10 "9 molar concentrations.
- the dosage in vivo may range, depending on the route of administration, between about 0.1 and 100 mg/kg.
- the cell adhesion inhibitory activity of these compounds may be measured by determining the concentration of inhibitor required to block the binding of VLA-4- expressing cells to fibronectin- or CS1 -coated plates.
- microtiter wells are coated with either fibronectin (containing the CS-1 sequence) or CS-1. If CS-1 is used, it must be conjugated to a carrier protein, such as bovine serum albumin, in order to bind to the wells.
- varying concentrations of the test compound are then added together with appropriately labeled, VLA-4-expressing cells. Alternatively, the test compound may be added first and allowed to incubate with the coated wells prior to the addition of the cells.
- the cells are allowed to incubate in the wells for at least 30 minutes. Following incubation, the wells are emptied and washed. Inhibition of binding is measured by quantitating the fluorescence or radioactivity bound to the plate for each of the various concentrations of test compound, as well as for controls containing no test compound.
- VLA-4-expressing cells that may be utilized in this assay include Ramos cells, Jurkat cells, A375 melanoma cells, as well as human peripheral blood lymophocytes (PBLs).
- the cells used in this assay may be fluorescently or radioactively labeled.
- a direct binding assay may also be employed to quantitate the inhibitory activity of the compounds of this invention.
- a VCAM-lgG fusion protein containing the first two immunoglobin domains of VCAM (D1D2) attached above the hinge region of an lgG1 molecule (VCAM 2D-lgG) is conjugated to a marker enzyme, such as alkaline phosphatase (AP).
- AP alkaline phosphatase
- VCAM-lgG enzyme conjugate is then placed in the wells of a multi-well filtration plate, such as that contained in the Millipore Multiscreen Assay System (Millipore Corp., Bedford, MA). Varying concentrations of the test inhibitory compound are then added to the wells followed by addition of VLA-4-expressing cells. The cells, compound and VCAM-lgG enzyme conjugate are mixed together and allowed to incubate at room temperature. Following incubation, the wells are vacuum drained, leaving behind the cells and any bound VCAM. Quantitation of bound VCAM is determined by adding an appropriate colorimetric substrate for the enzyme conjugated to VCAM-lgG and determining the amount of reaction cell adhesion inhibitory activity.
- assays for other major groups of integrins i.e., ⁇ 2 and ⁇ 3, as well as other ⁇ 1 integrins, such as VLA-5, VLA-6 and ⁇ 4 ⁇ 7 are performed.
- these assays may be similar to the adhesion inhibition and direct binding assays described above, substituting the appropriate integrin-expressing cell and corresponding ligand.
- polymorphonuclear cells PMNs express ⁇ 2 integrins on their surface and bind to ICAM.
- ⁇ 3 integrins are involved in platelet aggregation and inhibition may be measured in a standard platelet aggregation assay.
- VLA-5 binds specifically to Arg-Gly-Asp sequences
- VLA-6 binds to laminin.
- the compounds of the invention may also be tested in the antigen-induced pulmonary eosinophilia assay in the mouse, as described below.
- mice Male B6D2F1/J mice are sensitized by i.p. injection of 0.5 mL alum-precipitated antigen containing 8 ⁇ g of ovalbumin (OVA) adsorbed to 2 mg of aluminum hydroxide gel in a saline vehicle. Five days later the mice are given a booster injection with OV A/alum. Control animals are sensitized with alum only. Ten mice are used for each group.
- OVA ovalbumin
- the antagonists are dissolved in 2% DMSO and 150 mM TRIS, pH 8.8. A solvent control is included for each experiment. Drugs are administered orally 30 minutes prior to OVA exposure, and 6 hour after the first OVA exposure. The % inhibition is calculated by the formula:
- Animals are sacrificed by CO 2 asphyxiation 24 hour after the first antigen challenge.
- the tracheas are exposed and cannulated.
- the lungs are lavaged with 0.6 mL buffer (Hanks buffered saline with 10 mM Hepes, 0.5% BSA and 10 U/mL heparin).
- the number of eosinophils in the lavage is assessed by counting the total number of leukocytes and the percentage of eosinophils for each sample.
- the immunosuppressive activity can also be determined in further animal models of T- cell mediated immune responses such as experimental allergic encephalomyelitis, Freund's adjuvant or collagen-induced arthritis, and models of graft vs. host reactions.
- the antiarthritic activity can be determined in the collagen II induced arthritis in mice model.
- DBA/1 LacJ female mice, 6-8 weeks old are immunized with 100 ⁇ g of chicken type II collagen emulsified in Freund's complete adjuvant (FCA) by injecting at the base of the tail on day 0.
- FCA Freund's complete adjuvant
- the immune response to type II collagen is boosted with a subcutaneous injection of 200 ⁇ g LPS solubilized in PBS.
- Paws are examined on days 17, 21 , 24, 28, 31 , 35, and 42 and scored for the symptoms of arthritis based on criteria such as inflammation, swelling and ankylosis.
- Compounds are suspended in corn starch vehicle and administered orally to mice once daily beginning on day 1 and continuing through day 42.
- Protection against transplant rejection can be determined in the following mouse tail skin transplantation procedure.
- mice are anesthetized with tribromoethanol i.p. injection according to the method of Papaioannou and Fox (Lab. Animal Science 1993;43:189-92).
- the mouse tail skin transplant rejection model is a modification of that described by Baily and Usama, (Transpl. Bull. 1960;7:424-5).
- Four skin grafts per mouse are exchanged between strains [C57BIJ10-SnJ (H-2K b ) and B10.BR/SgSnJ (H-2K k )] at surgery (dO). Fitted skin grafts are made with a scalpel, and tail graft sites are protected with tubing for 2 days following surgery. Mouse tails are evaluated at day 7 for missing grafts.
- the compounds of the invention can be prepared by adaptation of previously reported synthetic methodology, e.g., in International Application WO 96/22966 (designating the U.S. and which is incorporated herein by reference) as described below and illustrated in the examples.
- the condensation is carried out according to methodology well known in the art for amide formation, e.g. in the presence of a condensing agent such as 1-[3- (dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride and a base, such as diisopropylethylamine, in an inert solvent (such as methylene chloride), preferably at room temperature.
- a condensing agent such as 1-[3- (dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride and a base, such as diisopropylethylamine, in an inert solvent (such as methylene chloride), preferably at room temperature.
- the starting materials of formula IV such as optionally substituted phenylureidophenylacetic acids, are in turn known in the art or are prepared according to methods known in the art, e.g. by, for example, condensing a p-aminophenylacetic acid ester with the appropriate aryl isocyanate to obtain the corresponding phenylureidophenylacetic acid ester and hydrolyzing the resulting ester.
- the starting materials of formula V are in turn prepared by reacting a compound of the formula
- R ⁇ and R 2 have meaning as defined hereinabove and L is a leaving group, such as halo or (alkyl or aryl)-sulfonyloxy, in the presence of a base, such as triethylamine,
- Y and Alk have meaning as defined hereinabove, under conditions well-known in the art, to obtain a starting material of formula V in protected form (e.g. as an alkyl ester).
- Hydrolysis e.g. with base, such as aqueous lithium hydroxide, gives a starting material of formula V.
- base such as aqueous lithium hydroxide
- the above-mentioned reactions are carried out according to standard methods, in the presence or absence of diluent, preferably such as are inert to the reagents and are solvents thereof, of catalysts, condensing or said other agents respectively and/or inert atmospheres, at low temperatures, room temperature or elevated temperatures (preferably at or near the boiling point of the solvents used), and at atmospheric or super-atmospheric pressure.
- diluent preferably such as are inert to the reagents and are solvents thereof, of catalysts, condensing or said other agents respectively and/or inert atmospheres, at low temperatures, room temperature or elevated temperatures (preferably at or near the boiling point of the solvents used), and at atmospheric or super-atmospheric pressure.
- diluent preferably such as are inert to the reagents and are solvents thereof, of catalysts, condensing or said other agents respectively and/or inert atmospheres, at low temperatures, room temperature or elevated temperatures (preferably at or near
- the invention also relates to any novel starting materials and processes for their manufacture.
- any mixtures of final products or intermediates obtained can be separated on the basis of the physico-chemical differences of the constituents, in known manner, into the pure final products or intermediates, for example by chromatography, distillation, fractional crystallization, or by formation of a salt if appropriate or possible under the circumstances.
- the compounds of the invention or intermediates can also be obtained in the form of their hydrates, or include other solvents used for their crystallization.
- Some of the compounds described herein contain one or more asymmetric centers and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms which may be defined in terms of absolute stereochemistry as (R) or (S), or as (D) or (L) for amino acids.
- the present invention is meant to include all such possible diastereomers as well as their racemic and optically pure forms.
- Optically active (R) and (S), or (D) and (L), isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.
- the compounds described herein contain olefinic double bonds or other centers of geometric asymmetry, and unless specified otherwise, it is intended to include both E and Z geometric isomers. Likewise, all tautomeric forms are intended to be included.
- the invention further relates to pharmaceutical compositions suitable for enteral, such as oral or rectal, transdermal, intranasal, topical ocular and parenteral administration to mammals including man, which are useful as VLA-4 antagonists and for the treatment of disorders responsive thereto, comprising an effective amount of a pharmacologically active compound of the invention, alone or in combination, with one or more pharmaceutically acceptable carriers.
- enteral such as oral or rectal, transdermal, intranasal, topical ocular and parenteral administration to mammals including man, which are useful as VLA-4 antagonists and for the treatment of disorders responsive thereto, comprising an effective amount of a pharmacologically active compound of the invention, alone or in combination, with one or more pharmaceutically acceptable carriers.
- compositions include compositions suitable for oral, rectal, topical (including transdermal devices, aerosols, creams, ointments, lotions, and dusting powders), parenteral (including subcutaneous, intramuscular, and intravenous), ocular (ophthalmic), pulmonary (nasal or buccal inhalation), or nasal administration; the most suitable route in any given case will depend largely on the nature and severity of the condition being treated and on the nature of the active ingredient.
- the compounds of the invention may be conveniently presented in a unit dosage form prepared by any of the methods well known in the art of pharmacy.
- compounds of the invention may be administered orally, for example in tablet form, or by inhalation, for example in aerosol or other atomisable formulations or in dry powder formulations, using an appropriate inhalation device such as those known in the art.
- inhalation for example in aerosol or other atomisable formulations or in dry powder formulations, using an appropriate inhalation device such as those known in the art.
- the compounds of the invention may also be administered intranasally.
- the compounds of the invention may also be administered topically e.g. as an eye drop, gel, ointment and the like.
- a compound of the invention may be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques.
- the carrier may take a wide variety of forms depending on the nature of the preparation desired for administration, i.e., oral, parenteral, etc.
- any of the usual pharmaceutical media may be used, such as water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like in the case of oral liquid preparations (e.g., suspensions, elixirs, and solutions); or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, etc.
- solid oral preparations such as powders, capsules, and tablets.
- Solid oral preparations are generally preferred over liquid oral preparations. Because of their ease of administration, tablets and capsules are the preferred oral dosage unit form. If desired, tablets may be coated by standard aqueous or non-aqueous techniques.
- the compounds of the invention may be administered by controlled release means and devices, e.g. a transdermal therapeutic system.
- compositions of the present invention suitable for oral administration may be prepared as discrete units such as capsules, cachets, or tablets each containing a predetermined amount of the active ingredient in powder or granular form or as a solution or suspension in an aqueous or nonaqueous liquid or in an oil-in-water or water-in-oil emulsion.
- Such compositions may be prepared by any of the methods known in the art of pharmacy.
- the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers, finely divided solid carriers, or both and then, if necessary, shaping the product into the desired form.
- a tablet may be prepared by compression or molding, optionally with one or more accessory ingredients.
- Compressed tablets may be prepared by compressing in a suitable machine the active ingredient in a free-flowing form such as powder or granule optionally mixed with a binder, lubricant, inert diluent, or surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent.
- compositions are preferably aqueous isotonic solutions emulsions or suspensions, and suppositories are advantageously prepared from fatty emulsions or suspensions.
- Said compositions may be sterilized and/or contain adjuvants, such as preserving, stabilizing, wetting or emulsifying agents, solution promoters, salts for regulating the osmotic pressure and/or buffers. In addition, they may also contain other therapeutically valuable substances.
- Said compositions are prepared according to conventional mixing, granulating or coating methods, respectively, and contain about 0.1 to 75%, preferably about 1 to 50% of the active ingredient.
- transdermal devices are in the form of a bandage comprising a backing member, optionally a reservoir containing the compound optionally with carriers, optionally a rate controlling barrier to deliver the compound of the skin of the host at a controlled and predetermined rate over a prolonged period of time, and an adhesive layer to secure the device to the skin.
- compositions containing a compound of this invention may also comprise an additional agent selected from the group consisting of e.g. cortiocosteroids, bronchodilators, antiasthmatics (mast cell stabilizers), anti-inflammatories, antirheumatics, immunosuppressants, antimetabolites, immunonodulators, antipsoriatics, and antidiabetics.
- additional agent selected from the group consisting of e.g. cortiocosteroids, bronchodilators, antiasthmatics (mast cell stabilizers), anti-inflammatories, antirheumatics, immunosuppressants, antimetabolites, immunonodulators, antipsoriatics, and antidiabetics.
- specific compounds that can be used in combination include cyclosporine, FK-506, and rapamycin (immunosuppressants); cyclophosphamide and methotrexate (antimetabolites); and interferons (i
- the invention further relates to a method of inhibiting VLA-4 activity in mammals and treating or preventing diseases and conditions responsive thereto described herein, e.g. autoimmune disorders, rejection of transplantation, psoriasis or respiratory diseases which comprises administering to a mammal in need thereof an effective amount of a compound of the invention or of a pharmaceutical composition comprising a said compound in combination with one or more pharmaceutically acceptable carriers.
- diseases and conditions responsive thereto described herein e.g. autoimmune disorders, rejection of transplantation, psoriasis or respiratory diseases
- a particular embodiment thereof relates to a method of inhibiting VLA-4 dependent cell adhesion in mammals which comprises administering to a mammal in need thereof a correspondingly effective inhibiting amount of a compound of the invention or of a said compound in combination with one or more pharmaceutically acceptable carriers.
- the prophylactic or therapeutic dose of the compounds of the invention will vary with the nature and severity of the condition to be treated and with the particular compound of the invention and its route of administration.
- the daily dose lies in the range of 0.05 to 50 mg/kg body weight of a mammal, preferably in the range of 0.1 to 10 mg/kg, in single or divided doses. In some cases, it may be necessary to use doses outside these ranges.
- a unit dosage for a mammal of about 50 to 70 kg may typically contain between about 1 and 150 mg of the active ingredient.
- EDAC 1 -[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride
- TRIS tris(hydroxymethyl)aminomethane
- the mixture is reduced to dryness and flash chromatographed using 150 g silica gel (230-400 mesh), and 1 to 4% CH 3 OH/CH 2 CI 2 as eluant to yield methyl ⁇ -amino- ⁇ -(3,4-dimethoxy-phenyl) propionate (product C) as a yellow oil.
- the mixture is reduced to dryness and flash chromatographed using 30 g silica gel, Merck, grade 9385 (230-400 mesh, 60 A) using 25% ethyl acetate/75% hexanes as eluant, to yield methyl ⁇ -(3,4- dimethyoxyphenyl)- ⁇ -(bromoacetylamino) propionate (product D) as a thick yellow oil.
- Example 1 Similarly to the procedure of Example 1 are prepared the following compounds (as the indicated enantiomers if in optically active form).
- the starting material is prepared as follows:
- Product C is condensed with product D of Example 1 , and the resulting ester is hydrolyzed (similarly to steps 6 and 7 of Example 1) to the title product which is purified by flash chromatography eluting with 10% CH 3 OH in CH 2 CI 2 ; m.p. 102-106° dec.
Abstract
Description
Claims
Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP00990650A EP1235793A2 (en) | 1999-12-07 | 2000-12-05 | Vla-4 integrin antagonists |
CA002388686A CA2388686A1 (en) | 1999-12-07 | 2000-12-05 | Vla-4 integrin antagonists |
AU30064/01A AU3006401A (en) | 1999-12-07 | 2000-12-05 | Vla-4 integrin antagonists |
JP2001543494A JP2003516380A (en) | 1999-12-07 | 2000-12-05 | VLA-4 integrin antagonist |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US45567099A | 1999-12-07 | 1999-12-07 | |
US09/455,670 | 1999-12-07 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2001042192A2 true WO2001042192A2 (en) | 2001-06-14 |
WO2001042192A3 WO2001042192A3 (en) | 2002-01-10 |
Family
ID=23809780
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/EP2000/012226 WO2001042192A2 (en) | 1999-12-07 | 2000-12-05 | Vla-4 integrin antagonists |
Country Status (5)
Country | Link |
---|---|
EP (1) | EP1235793A2 (en) |
JP (1) | JP2003516380A (en) |
AU (1) | AU3006401A (en) |
CA (1) | CA2388686A1 (en) |
WO (1) | WO2001042192A2 (en) |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6969728B2 (en) | 2000-05-12 | 2005-11-29 | Genzyme Corporation | Modulators of TNF-α signaling |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1998004247A1 (en) * | 1996-07-25 | 1998-02-05 | Biogen, Inc. | Cell adhesion inhibitors |
WO1999037605A1 (en) * | 1998-01-23 | 1999-07-29 | Novartis Ag | Vla-4 antagonists |
US5936065A (en) * | 1993-12-06 | 1999-08-10 | Cytel Corporation | CS-1 peptidomimetics, compositions and methods of using the same |
-
2000
- 2000-12-05 JP JP2001543494A patent/JP2003516380A/en active Pending
- 2000-12-05 AU AU30064/01A patent/AU3006401A/en not_active Abandoned
- 2000-12-05 WO PCT/EP2000/012226 patent/WO2001042192A2/en not_active Application Discontinuation
- 2000-12-05 EP EP00990650A patent/EP1235793A2/en not_active Withdrawn
- 2000-12-05 CA CA002388686A patent/CA2388686A1/en not_active Abandoned
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5936065A (en) * | 1993-12-06 | 1999-08-10 | Cytel Corporation | CS-1 peptidomimetics, compositions and methods of using the same |
WO1998004247A1 (en) * | 1996-07-25 | 1998-02-05 | Biogen, Inc. | Cell adhesion inhibitors |
WO1999037605A1 (en) * | 1998-01-23 | 1999-07-29 | Novartis Ag | Vla-4 antagonists |
Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6969728B2 (en) | 2000-05-12 | 2005-11-29 | Genzyme Corporation | Modulators of TNF-α signaling |
US7034031B2 (en) | 2000-05-12 | 2006-04-25 | Genzyme Corporation | Modulators of TNF-α signaling |
US8518999B2 (en) | 2000-05-12 | 2013-08-27 | Genzyme Corporation | Modulators of TNF-αsignaling |
US8921547B2 (en) | 2000-05-12 | 2014-12-30 | Genzyme Corporation | Modulators of TNF-α signaling |
US9579325B2 (en) | 2000-05-12 | 2017-02-28 | Genzyme Corporation | Modulators of TNF-α signaling |
Also Published As
Publication number | Publication date |
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AU3006401A (en) | 2001-06-18 |
CA2388686A1 (en) | 2001-06-14 |
JP2003516380A (en) | 2003-05-13 |
WO2001042192A3 (en) | 2002-01-10 |
EP1235793A2 (en) | 2002-09-04 |
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