+

WO1999025685A1 - Derives de 4-piperidine carboxamides 4-substitues - Google Patents

Derives de 4-piperidine carboxamides 4-substitues Download PDF

Info

Publication number
WO1999025685A1
WO1999025685A1 PCT/US1998/024513 US9824513W WO9925685A1 WO 1999025685 A1 WO1999025685 A1 WO 1999025685A1 US 9824513 W US9824513 W US 9824513W WO 9925685 A1 WO9925685 A1 WO 9925685A1
Authority
WO
WIPO (PCT)
Prior art keywords
mpupa
independently selected
ioalkyl
optionally substituted
benzyl
Prior art date
Application number
PCT/US1998/024513
Other languages
English (en)
Inventor
Stephen E. Delaszlo
William K. Hagmann
Original Assignee
Merck & Co., Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB9727214.0A external-priority patent/GB9727214D0/en
Application filed by Merck & Co., Inc. filed Critical Merck & Co., Inc.
Priority to AU14150/99A priority Critical patent/AU1415099A/en
Publication of WO1999025685A1 publication Critical patent/WO1999025685A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D211/60—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D211/62—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals attached in position 4
    • C07D211/64—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals attached in position 4 having an aryl radical as the second substituent in position 4
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D211/00—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings
    • C07D211/04—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
    • C07D211/06—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
    • C07D211/36—Heterocyclic compounds containing hydrogenated pyridine rings, not condensed with other rings with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D211/60—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
    • C07D211/62—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals attached in position 4

Definitions

  • the present invention relates to novel 4-substituted-4- piperidinecarboxamide derivatives which are useful for the inhibition and prevention of leukocyte adhesion and leukocyte adhesion-mediated pathologies.
  • This invention also relates to compositions containing such compounds and methods of treatment using such compounds.
  • Many physiological processes require that cells come into close contact with other cells and/or extracellular matrix. Such adhesion events may be required for cell activation, migration, proliferation and differentiation.
  • Cell-cell and cell-matrix interactions are mediated through several families of cell adhesion molecules (CAMs) including the selectins, integrins, cadherins and immunoglobulins.
  • CAMs play an essential role in both normal and pathophysiological processes.
  • the integrin superfamily is made up of structurally and functionally related glycoproteins consisting of and ⁇ heterodimeric, transmembrane receptor molecules found in various combinations on nearly every mammalian cell type, (for reviews see: E. C. Butcher, Cell. £7, 1033 (1991); T. A. Springer, Cell, 76, 301 (1994); D. Cox et al., "The Pharmacology of the Integrins.” Medicinal Research Rev. 14. 195 (1994) and V. W.
  • VLA-4 very late antigen-4"; CD49d/CD29; or ⁇ 4 ⁇ i
  • VLA-4 very late antigen-4"; CD49d/CD29; or ⁇ 4 ⁇ i
  • CD49d/CD29 CD49d/CD29
  • ⁇ 4 ⁇ i is an integrin expressed on all leukocytes, except platelets and mature neutrophils, and is a key mediator of the cell-cell and cell-matrix interactions of leukocytes (see M. E. Hemler, "VLA Proteins in the Integrin Family: Structures, Functions, and Their Role on Leukocytes.” Ann. Rev. Immunol. 8, 365 (1990)).
  • VCAM-1 vascular cell adhesion molecule-1
  • FN fibronectin
  • VCAM-1 is a member of the Ig superfamily and is expressed in vivo on endothelial cells at sites of inflammation and on dendritic and macrophage-like cells.
  • VCAM-1 is produced by vascular endothelial cells in response to pro-inflammatory cytokines (See A. J. H. Gearing and W. Newman, "Circulating adhesion molecules in disease.”,
  • the CS-1 domain is a 25 amino acid sequence that arises by alternative splicing within a region of fibronectin.
  • a role for VLA-4/CS-1 interactions in inflammatory conditions has been proposed (see M. J. Elices, "The integrin ( 4 ⁇ (VLA-
  • ⁇ 4 ⁇ 7 (also referred to as LPAM-1 and oc ⁇ ) is an integrin expressed on leukocytes and is a key mediator of leukocyte trafficking and homing in the gastrointestinal tract (see C M. Parker et al., Proc. Natl. Acad. Sci. USA, fg, 1924 (1992)).
  • the ligands for ⁇ 4 ⁇ 7 include mucosal addressing cell adhesion molecule-1 (MadCAM-1) and, upon activation of ⁇ 4 ⁇ 7, VCAM-1 and fibronectin (Fn).
  • MadCAM-1 is a member of the Ig superfamily and is expressed in vivo on endothelial cells of gut-associated mucosal tissues of the small and large intestine ("Peyer's Patches") and lactating mammary glands. (See M. J. Briskin et al., Nature. 363. 461 (1993); A. Hamann et al., J. Immunol.. 152. 3282 (1994)). MadCAM-1 can be induced in vitro by proinflammatory stimuli (See E. E. Sikorski et al. J. Immunol.. 151. 5239 (1993)). MadCAM-1 is selectively expressed at sites of lymphocyte extravasation and specifically binds to the integrin, ( 4 ⁇ 7.
  • Neutralizing anti- ⁇ 4 antibodies or blocking peptides that inhibit the interaction between VLA-4 and/or ( 4 ⁇ 7 and their ligands have proven efficacious both prophylactically and therapeutically in several animal models of disease, including i) experimental allergic encephalomyelitis, a model of neuronal demyelination resembling multiple sclerosis (for example, see T. Yednock et al., "Prevention of experimental autoimmune encephalomyelitis by antibodies against 04 ⁇ integrin.” Nature. 356, 6J3 (1993) and E. Keszthelyi et al., "Evidence for a prolonged role of 0.4 integrin throughout active experimental allergic encephalomyelitis.” Neurology.
  • VLA-4 interactions in other diseases including rheumatoid arthritis; various melanomas, carcinomas, and sarcomas; inflammatory lung disorders; atherosclerotic plaque formation; restenosis; and circulatory shock (for examples, see A. A. Postigo et al., "The ⁇ 4 ⁇ /VCAM-l adhesion pathway in physiology and disease.”, Res. Immunol.. 144. 723 (1994) and J.-X. Gao and A. C. Issekutz, "Expression of VCAM-1 and VLA-4 dependent T-lymphocyte adhesion to dermal fibroblasts stimulated with proinflammatory cytokines.” Immunol. .89, 375 (1996)).
  • a humanized monoclonal antibody for example, see A. A. Postigo et al., "The ⁇ 4 ⁇ /VCAM-l adhesion pathway in physiology and disease.”, Res. Immunol.. 144. 723 (1994) and J.-X
  • VLA-4 (Antegren® Athena Neurosciences/Elan ) against VLA-4 in clinical development for the treatment of "flares” associated with multiple sclerosis and a humanized monoclonal antibody (ACT-1® LeukoSite) against 0-4 ⁇ 7 in clinical development for the treatment of inflammatory bowel disease.
  • ACT-1® LeukoSite a humanized monoclonal antibody against 0-4 ⁇ 7 in clinical development for the treatment of inflammatory bowel disease.
  • peptidyl antagonists of VLA-4 have been described (D. Y. Jackson et al., "Potent ⁇ 4 ⁇ l peptide antagonists as potential anti-inflammatory agents", J. Med. Chem.. 0, 3359 (1997); H. N. Shroff et al., "Small peptide inhibitors of ⁇ 4 ⁇ 7 mediated MadCAM-1 adhesion to lymphocytes", Bioorg. Med. Chem. Lett.. 6, 2495 (1996); US 5,510,332, WO97/0309
  • VLA-4- and ⁇ 4 ⁇ 7-dependent cell adhesion that have improved pharmacokinetic and pharmacodynamic properties such as oral bioavailability and significant duration of action.
  • Such compounds would prove to be useful for the treatment, prevention or suppression of various pathologies mediated by VLA-4 and ⁇ 4 ⁇ 7 binding and cell adhesion and activation.
  • the compounds of the present invention are antagonists of the VLA-4 integrin ("very late antigen-4"; CD49d/CD29; or ⁇ 4 ⁇ l) and/or the ⁇ 4 ⁇ 7 integrin (LPAM-1 and ( 4 ⁇ p), thereby blocking the binding of VLA-4 to its various ligands, such as VCAM-1 and regions of fibronectin and/or 4 ⁇ 7 to its various ligands, such as MadCAM-1, VCAM-1 and fibronectin.
  • VLA-4 integrin very late antigen-4"; CD49d/CD29; or ⁇ 4 ⁇ l
  • LPAM-1 and ( 4 ⁇ p) ⁇ 4 ⁇ 7 integrin
  • these antagonists are useful in inhibiting cell adhesion processes including cell activation, migration, proliferation and differentiation.
  • VLA-4 and/or ⁇ 4 ⁇ 7 binding and cell adhesion and activation are useful in the treatment, prevention and suppression of diseases mediated by VLA-4 and/or ⁇ 4 ⁇ 7 binding and cell adhesion and activation, such as multiple sclerosis, asthma, allergic rhinitis, allergic conjunctivitis, inflammatory lung diseases, rheumatoid arthritis, septic arthritis, type I diabetes, organ transplantation, restenosis, autologous bone marrow transplantation, inflammatory sequelae of viral infections, myocarditis, inflammatory bowel disease including ulcerative colitis and Crohn's disease, certain types of toxic and immune-based nephritis, contact dermal hypersensitivity, psoriasis, tumor metastasis, and atherosclerosis.
  • diseases mediated by VLA-4 and/or ⁇ 4 ⁇ 7 binding and cell adhesion and activation such as multiple sclerosis, asthma, allergic rhinitis, allergic conjunctivitis, inflammatory lung diseases, rheum
  • the present invention provides novel compounds of Formula I
  • Rl is 1) Ci-ioalkyl
  • R3 is 1) hydrogen
  • Ci-i ⁇ alkyl optionally substituted with one to four substituents independently selected from R a
  • Cy optionally substituted with one to four substituents independently selected from R D , R4 is 1) hydrogen
  • R 7 is 1) hydrogen
  • R" is 1) a group selected from R a ,
  • R d and R e are independently selected from the group consisting of
  • Cy Ci-ioalkyl wherein alkyl, alkenyl, alkynyl and Cy is optionally substituted with one to four substituents independently selected from R c ; or R d and R e together with the atoms to which they are attached form a heterocyclic ring of 5 to 7 members containing 0-2 additional heteroatoms independently selected from oxygen, sulfur and nitrogen; Rf and RS are independently selected from hydrogen, Ci-ioalkyl, Cy and Cy Ci-ioalkyl; or R and RS together with the carbon to which they are attached form a ring of 5 to 7 members containing 0-2 heteroatoms independently selected from oxygen, sulfur and nitrogen; R n is 1) hydrogen, 2) Ci-ioalkyl,
  • alkyl, alkenyl, and alkynyl are optionally substituted with one to four substituents independently selected from R a ; and aryl and heteroaryl are each optionally substituted with one to four substituents independently selected from R D ;
  • alkyl, alkenyl, alkynyl and aryl are each optionally substituted with one to four substituents independently selected from R c ;
  • Cy and Cyl are indep endently selected 1) cycloalkyl,
  • Rl is Ci-ioalkyl, Cy or Cy-Ci-ioalkyl wherin alkyl and Cy are optionally substituted as provided above.
  • Rl Cy is preferably aryl optionally substituted with one or two substituents independently selected from Rb.
  • Another subset of compounds of formula I are compounds wherein L is SO2 or C(O). Preferably L is SO2.
  • R2 is Ci-i ⁇ alkyl, Cy-Ci-ioalkyl or Cy each optionally substituted as provided above.
  • R2 Cy is preferably aryl.
  • L is 1) -C(O)- or
  • X is 1) a bond or
  • Rl is 1) Ci-ioalkyl
  • R2 is 1) a group selected from Rl;
  • R4 is 1) hydrogen
  • R 6 is 1) hydrogen, or
  • R ⁇ is 1) hydrogen; all the other variables are as defined above under formula I.
  • Another preferred embodiment of compounds of formula I are compounds of formula lb: lb X is 1) a bond or
  • Rl is 1) Ci-ioalkyl
  • I c X is 1) a bond or
  • R5 is 1) hydrogen, 2) Ci-ioalkyl,
  • Cy-(Cyl) p -Ci-ioalkyl wherein alkyl is optionally substituted with one to two substituents independently selected from R a ; and Cy and Cyl are optionally substituted with one to two substituents independently selected from R D ; Cy and Cyl are are independently
  • Id X is 1) a bond or
  • Alkyl as well as other groups having the prefix “alk”, such as alkoxy, alkanoyl, means carbon chains which may be linear or branched or combinations thereof.
  • alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, sec- and tert-butyl, pentyl, hexyl, heptyl, octyl, nonyl, and the like.
  • alkenyl means carbon chains which contain at least one carbon-carbon double bond, and which may be linear or branched or combinations thereof. Examples of alkenyl include vinyl, allyl, isopropenyl, pentenyl, hexenyl, heptenyl, 1-propenyl, 2-butenyl, 2- methyl-2-butenyl, and the like.
  • Alkynyl means carbon chains which contain at least one carbon-carbon triple bond, and which may be linear or branched or combinations thereof. Examples of alkynyl include ethynyl, propargyl, 3-methyl-l-pentynyl, 2-heptynyl and the like.
  • Cycloalkyl means mono- or bicyclic saturated carbocyclic rings, each of which having from 3 to 10 carbon atoms. The term also inccludes monocyclic ring fused to an aryl group in which the point of attachment is on the non-aromatic portion. Examples of cycloalkyl include cyclopropyl, cyclopentyl, cyclohexyl, cycloheptyl, tetrahydronaphthyl, decahydronaphthyl, indanyl, and the like.
  • Aryl means mono- or bicyclic aromatic rings containing only carbon atoms.
  • the term also includes aryl group fused to a monocyclic cycloalkyl or monocyclic heterocyclyl group in which the point of attachment is on the aromatic portion.
  • aryl include phenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3- dihydrobenzofuranyl, benzopyranyl, 1,4-benzodioxanyl, and the like.
  • Heteroaryl means a mono- or bicyclic aromatic ring containing at least one heteroatom selected from N, O and S, with each ring containing 5 to 6 atoms.
  • heteroaryl include pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, triazinyl, thienyl, pyrimidyl, pyridazinyl, pyrazinyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, benzothiophenyl, furo(2,3-b)pyridyl, quinolyl, indolyl, isoquinolyl, and the like.
  • Heterocyclyl means mono- or bicyclic saturated rings containing at least one heteroatom selected from N, S and O, each of said ring having from 3 to 10 atoms.
  • the term also includes monocyclic heterocycle fused to an aryl or heteroaryl group in which the point of attachment is on the non-aromatic portion.
  • heterocyclyl include pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, 2,3- dihydrofuro(2,3-b)pyridyl, benzoxazinyl, tetrahydrohydroquinolinyl, tetrahydroisoquinolinyl, dihydroindolyl, and the like.
  • Halogen includes fluorine, chlorine, bromine and iodine.
  • Compounds of Formula I contain one or more asymmetric centers and can thus occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The present invention is meant to comprehend all such isomeric forms of the compounds of Formula I.
  • tautomers Some of the compounds described herein may exist with different points of attachment of hydrogen, referred to as tautomers. Such an example may be a ketone and its enol form known as keto-enol tautomers. The individual tautomers as well as mixture thereof are encompassed with compounds of Formula I.
  • Compounds of the Formula I may be separated into diastereoisomeric pairs of enantiomers by, for example, fractional crystallization from a suitable solvent, for example methanol or ethyl acetate or a mixture thereof.
  • a suitable solvent for example methanol or ethyl acetate or a mixture thereof.
  • the pair of enantiomers thus obtained may be separated into individual stereoisomers by conventional means, for example by the use of an optically active acid as a resolving agent.
  • any enantiomer of a compound of the general Formula I or la may be obtained by stereospecif c synthesis using optically pure starting materials or reagents of known configuration.
  • pharmaceutically acceptable salts refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids
  • Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic salts, manganous, potassium, sodium, zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts.
  • Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-dibenzylethylenediamine, 2-diethylaminoethanol, 2- dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethyl- morpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.
  • salts may be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids.
  • acids include acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, ucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p- toluenesulfonic acid, and the like.
  • Particularly preferred are citric, hydrobromic, hydrochloric, maleic, phosphoric, sulfuric, and tartaric acids.
  • Another aspect of the present invention provides a method for the treatment (including prevention, alleviation, amelioration or suppression) of diseases or disorders or symptoms mediated by VLA-4 and/or 4 ⁇ 7 binding and cell adhesion and activation, which comprises administering to a mammal an effective amount of a compound of Formula I.
  • Such diseases, disorders, conditions or symptoms are for example (1) multiple sclerosis, (2) asthma, (3) allergic rhinitis, (4) allergic conjunctivitis, (5) inflammatory lung diseases, (6) rheumatoid arthritis, (7) septic arthritis, (8) type I diabetes, (9) organ transplantation rejection, (10) restenosis, (11) autologous bone marrow transplantation, (12) inflammatory sequelae of viral infections, (13) myocarditis, (14) inflammatory bowel disease including ulcerative colitis and Crohn's disease, (15) certain types of toxic and immune-based nephritis, (16) contact dermal hypersensitivity, (17) psoriasis, (18) tumor metastasis, (19) hepatitis, and (20) atherosclerosis.
  • prophylactic or therapeutic dose of a compound of Formula I will, of course, vary with the nature of the severity of the condition to be treated and with the particular compound of Formula I and its route of administration. It will also vary according to the age, weight and response of the individual patient. In general, the daily dose range lie within the range of from about 0.001 mg to about 100 mg per kg body weight of a mammal, preferably 0.01 mg to about 50 mg per kg, and most preferably 0.1 to 10 mg per kg, in single or divided doses. On the other hand, it may be necessary to use dosages outside these limits in some cases.
  • a suitable dosage range is from about 0.001 mg to about 25 mg (preferably from 0.01 mg to about 1 mg) of a compound of Formula I per kg of body weight per day and for cytoprotective use from about 0.1 mg to about 100 mg (preferably from about 1 mg to about 100 mg and more preferably from about 1 mg to about 10 mg) of a compound of Formula I per kg of body weight per day.
  • a suitable dosage range is, e.g.
  • ophthalmic preparations for ocular administration comprising 0.001-1% by weight solutions or suspensions of the compounds of Formula I in an acceptable ophthalmic formulation may be used.
  • compositions which comprises a compound of Formula I and a pharmaceutically acceptable carrier.
  • composition is intended to encompass a product comprising the active ingredient(s), and the inert ingredient(s) (pharmaceutically acceptable excipients) that make up the carrier, as well as any product which results, directly or indirectly, from combination, complexation or aggregation of any two or more of the ingredients, or from dissociation of one or more of the ingredients, or from other types of reactions or interactions of one or more of the ingredients.
  • the pharmaceutical compositions of the present invention encompass any composition made by admixing a compound of Formula I, additional active ingredient(s), and pharmaceutically acceptable excipients.
  • Any suitable route of administration may be employed for providing a mammal, especially a human with an effective dosage of a compound of the present invention.
  • a mammal especially a human with an effective dosage of a compound of the present invention.
  • suitable route of administration for example, oral, rectal, topical,
  • Dosage forms include tablets, troches, dispersions, suspensions, solutions, capsules, creams, ointments, aerosols, and the like.
  • compositions of the present invention comprise a compound of Formula I as an active ingredient or a pharmaceutically acceptable salt thereof, and may also contain a pharmaceutically acceptable carrier and optionally other therapeutic ingredients.
  • pharmaceutically acceptable salts refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids including inorganic bases or acids and organic bases or acids.
  • the compositions include compositions suitable for oral, rectal, topical, parenteral (including subcutaneous, intramuscular, and intravenous), ocular (ophthalmic), pulmonary (nasal or buccal inhalation), or nasal administration, although the most suitable route in any given case will depend on the nature and severity of the conditions being treated and on the nature of the active ingredient. They may be conveniently presented in unit dosage form and prepared by any of the methods well-known in the art of pharmacy.
  • the compounds of the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or nebulisers.
  • the compounds may also be delivered as powders which may be formulated and the powder composition may be inhaled with the aid of an insufflation powder inhaler device.
  • the preferred delivery system for inhalation is a metered dose inhalation (MDI) aerosol, which may be formulated as a suspension or solution of a compound of Formula I in suitable propellants, such as fluorocarbons or hydrocarbons.
  • MDI metered dose inhalation
  • Suitable topical formulations of a compound of formula I include transdermal devices, aerosols, creams, ointments, lotions, dusting powders, and the like.
  • the compounds of Formula I can 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 form of preparation desired for administration, e.g., oral or parenteral (including intravenous).
  • any of the usual pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents and the like in the case of oral liquid preparations, such as, for example, suspensions, elixirs and solutions; or carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents and the like in the case of oral solid preparations such as, for example, powders, capsules and tablets, with the solid oral preparations being preferred over the liquid preparations. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. If desired, tablets may be coated by standard aqueous or nonaqueous techniques.
  • compositions of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient, as a powder or granules or as a solution or a suspension in an aqueous liquid, a non-aqueous liquid, an oil-in-water emulsion or a water-in-oil liquid emulsion.
  • compositions may be prepared by any of the methods of pharmacy but all methods include the step of bringing into association the active ingredient with the carrier which constitutes one or more necessary ingredients.
  • the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product into the desired presentation.
  • 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 granules, optionally mixed with a binder, lubricant, inert diluent, 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. Desirably, each tablet contains from about 1 mg to about 500 mg of the active ingredient and each cachet or capsule contains from about 1 to about 500 mg of the active ingredient.
  • Compounds of Formula I may be used in combination with other drugs that are used in the treatment/prevention/suppression or amelioration of the diseases or conditions for which compounds of Formula I are useful. Such other drugs may be administered, by a route and in an amount commonly used therefor, contemporaneously or sequentially with a compound of Formula I.
  • a pharmaceutical composition containing such other drugs in addition to the compound of Formula I is preferred.
  • the pharmaceutical compositions of the present invention include those that also contain one or more other active ingredients, in addition to a compound of Formula I. Examples of other active ingredients that may be combined with a compound of Formula I, either administered separately or in the same pharmaceutical compositions, include, but are not limited to:
  • VLA-4 antagonists such as those described in US 5,510,332, WO97/03094, WO97/02289, WO96/40781, WO96/22966, WO96/20216, WO96/01644, WO96/06108, WO95/15973, WO96/31206 and WO98/42656;
  • steroids such as beclomethasone, methylprednisolone, betamethasone, prednisone, dexamethasone, and hydrocortisone;
  • immunosuppressants such as cyclosporin, tacrolimus, rapamycin and other FK-506 type immunosuppressants;
  • antihistamines Hl- histamine antagonists
  • the weight ratio of the compound of the Formula I to the second active ingredient may be varied and will depend upon the effective dose of each ingredient. Generally, an effective dose of each will be used. Thus, for example, when a compound of the Formula I is combined with an NSAID the weight ratio of the compound of the
  • Formula I to the NSAID will generally range from about 1000:1 to about 1:1000, preferably about 200:1 to about 1:200. Combinations of a compound of the Formula I and other active ingredients will generally also be within the aforementioned range, but in each case, an effective dose of each active ingredient should be used.
  • the central piperidine scaffold may be prepared as outlined in Scheme 1 (for some examples of this sequence see Bioorganic and Medicinal Chemical Letters 7 (19), 2531, 1994).
  • the commercially available bis-2-chloroethyl amine hydrochloride A is protected at nitrogen by introduction of a protecting group such as benzyl, methyl, t- butoxycarbonyl (BOC), benzyloxycarbonyl (Cbz) in a manner known to one skilled in the art to give B.
  • BOC benzyloxycarbonyl
  • Cbz benzyloxycarbonyl
  • D may also be derived from protected piperidine- 4-carboxylic acid esters E by deprotonation with a strong base followed by alkylation with an alkyl group substituted with a leaving group.
  • Manipulation of the 'G' group of D in a manner known to one skilled in the art will provide the piperidine F ready for incorporation into compounds of Formula 1. It may be necessary to adjust the protecting group of the piperidine nitrogen, for example to FMOC (fluorenylmethylcarbonyl) for solid phase synthesis, this is well established for one skilled in the art.
  • the Fmoc-protected piperidine carboxylic acid derivative E is coupled to D employing standard peptide (in this instance, 2-(lH-benzotriazol-l-yl)- 1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), HOBt, and N,N-diisopropylethylamine (DIEA) in DMF to yield dipeptoid F.
  • HBTU 2-(lH-benzotriazol-l-yl)- 1,1,3,3-tetramethyluronium hexafluorophosphate
  • DIEA N,N-diisopropylethylamine
  • the Fmoc group is removed with piperidine in DMF to yield the free amine G.
  • the amine is then derivatized with the appropriate R L group by sulfonylation or acylation to give H as indicated on the scheme.
  • the product of this reaction may in itself, be converted further to other R 1 groups by utilization of functional group interconversions well known to one skilled in the art.
  • the final product is removed from the resin with strong acid (in this instance, trifluoroacetic acid (TFA) in the presence, or absence (depending on the sensitivity of the product) of thioanisole and dithiane) to yield compounds of the present invention I .
  • strong acid in this instance, trifluoroacetic acid (TFA) in the presence, or absence (depending on the sensitivity of the product) of thioanisole and dithiane
  • the amine is then derivatized with the appropriate R L group by sulfonylation or acylation as indicated on the scheme.
  • the product of this reaction may in itself, be converted further to other R 1 groups by utilization of functional group interconversions well know to one skilled in the art.
  • the final product is prepared by acid catalysed removal of the t-butyl ester by treatment with and acid such as TFA.
  • A CBz, FMOC, BOC
  • a biaryl moiety may be prepared as outlined in Scheme 4.
  • the Z group is protected by means known to one skilled in the art.
  • Substituted aryl or heteroaryl boronic acids are coupled to A in the presence of a palladium(O) reagent, such as tetrakis(triphenylphosphine)palladium under Suzuki conditions (N. Miyaura et al., Synth. Commun., 1981, 11, 513-519) to yieldB.
  • Tyrosine triflate starting materials are prepared by treatment of the tyrosine analog of A with triflic anhydride in pyridine. The Z group protecting group is then removed to give the corresponding compounds of Formula I.
  • the bromide or iodide can be converted into the desired boronic acid by treatment with an alkyllithium reagent in tetrahydrofuran at low temperature followed by addition of trimethyl or triisopropyl borate. Hydrolysis to the boronic acid can be effected by treatment of the intermediate with aqueous base and then acid.
  • Aryl boronates which may also be utilized in coupling reactions in place of aryl boronic acids may be prepared by palladium catalyzed boronation of aryl iodides and bromides as decribed in J. Org Chem , 1995, 60, 7508-7510.
  • the aryl coupling reaction may be performed by application of Stille-type carbon-carbon bond forming conditions (Scheme 5). (A.M. Echavarren and J.K. Stille, J. Am. Chem. Soc. 1987, 109, 5478-5486). .
  • the Z group is protected by means known to one skilled in the art.
  • the aryl bromide or iodide intermediate A is converted into its trimethyltin derivative B using hexamethylditin in the presence of palladium(O) and lithium chloride and then reacted with an appropriately substituted aryl or heteroaryl bromide, iodide, or triflate in the presence of a palladium reagent, such as tetrakis(triphenyl phosphine)palladium(O) or tris(dibenzylideneacetone)dipalladium(0), in a suitable solvent, such as toluene, dioxane, DMF, or l-methyl-2- pyrrolidinone, to give intermediate C.
  • a palladium reagent such as tetrakis(triphenyl phosphine)palladium(O) or tris(dibenzylideneacetone)dipalladium(0)
  • a suitable solvent such as toluene, dioxane, D
  • Aryl boronates may also be utilized in coupling reactions in place of aryl stannane. They may be prepared by palladium catalyzed boronation of aryl iodides and bromides as decribed in J. Org Chem , 1995, 60, 7508-7510. The resulting boronate may then be coupled to aryl bromide or iodide to provide C
  • Step C Preparation of N-BOC-4-n-butyl-4-piperidinecarboxylic acid.
  • 1.5 g (5 mmol) of the product of Step B was heated at 70° C over night in a mixture of 5 ml of 5M NaOH in water (25 mmol) and 5 ml of EtOH.
  • the reaction mixture was concentrated in vacuo and the aqueous residue was washed with ether.
  • the aqueous phase was acidified with 2N HC1 and extracted with EtOAc.
  • the organic phase was dried over MgSO 4 , filtered and concentrated in vacuo to provide the desired product.
  • Step D Preparation of N-FMOC-4-n-butyl-4-piperidinecarboxylic acid.
  • Step A Ethyl N-BOC-4-methyl-4-piperidinecarboxylate was prepared as described in Step B of Preparation 2 using iodomethane as the alkylating agent.
  • ⁇ -NMR CDC1 3 , 400Mhz: 1.18 (s, 3H); 1.25 (d,t, 3H); 1.31 (dt, 2H); 1.42 (s, 9H); 2.03 (bd, 2H); 2.95 (dt, 2H); 3.72 (m, 2H); 4.14 (q, 2H).
  • Step B N-BOC-4-methyl-4-piperidinecarboxylic acid was prepared as described in Step C of Preparation 2 using the product of Step A.
  • 1 H- NMR (CDC1 3 , 400MHz): 1.25 (s, 3H); 1.39 ( , 2H); 1.45 (s, 9H); 2.05 (bd, 2H); 3.05 (dt, 2H); 3.78 (bdt, 2H).
  • Step C Preparation of N-FMOC-4-methyl-4-piperidinecarboxylic acid was prepared as described in Step D of Preparation 2 utilizing the product of Step B as starting material, except as follows. After stirring the reaction mixture overnight in the presence of FMOCCl the mixture was concentrated in vacuo to remove dioxane. The aqueous phase was diluted with water and extracted with ether. The desired product was extracted into the ether phase (salt is soluble in ether). The combined ether and aqueous phase were acidified and extracted with ether. The combined ethereal phases were washed with brine, dried over MgSO 4 , filtered and concentrated in vacuo.
  • Step A Ethyl N-BOC-4-(4-fluorobenzyl)-4-piperidinecarboxylate was prepared as described in Step B of Preparation 2 using 4- bromomethylfluorobenzene as the alkylating agent.
  • ⁇ -NMR (CDC1 3 , 400MHz): 1.17 (t, 3H); 1.20-1.40 (m, 2H); 2.05 (bd, 2H); 2.76 (s, 2H), 2.72- 2.82 (m, 2H); 4.91 (bm, 2H); 4.08 (q, 2H); 6.89-7.00 (m, 4H).
  • Step B N-BOC-4-(4-fluorobenzyl)-4-piperidinecarboxylic acid was prepared as described in Step C of Preparation 2 using the product of Step A.
  • ⁇ -NMR CDC1 3 , 400MHz: 1.38-1.46 (m, 2H); 1.42 (s, 9H); 2.03 (bd, 2H); 2.82 (s, 2H); 2.85 (bt, 2H); 3.92 (bd, 2H); 6.93 (t, 2H); 7.04 (dd, 2H).
  • Step C N-FMOC-4-(4-fluorobenzyl)-4-piperidinecarboxylic acid was prepared as described in Step C, Preparation 3 utilizing the product of Step B as starting material.
  • Step A Loading of N-Fmoc-amino acid derivatives onto resins.
  • N-Fmoc-amino acids were loaded onto Tantagel-HMPB- amino acid resin to provide:
  • the resin was prepared in the following fashion: 25 grams of Tantagel amine resin (commercially available) was swollen in methylene chloride (150 ml). To this mixture was added 3.74 g of 4- hyrdroxymethyl-3-methoxy-phenoxybutyric acid (commercially available), 2.1 g of N-hydroxybenzotriazole (HOBt) and 1.88 g of diisopropyl carbodiimide. The mixture was stirred over night at room temperature, filtered and the residue was washed with methylene chloride (3 x 150 ml), MeOH ( 3 x 150 ml), methylene chloride (3 x 150 ml) and ether ( 3 x 100 ml). The residue was dried in vacuo to give the handle derivatized resin ready for loading with the appropriate amino acid.
  • the resin (2.0 g, 0.54 mmol) was swollen in 50% THF/CH 2 C1 2 and suspended in 5 ml of the same solvent. 5 mg of dimethylaminopyridine (DMAP) was added followed by 1.0 mmol of the amino acid N-protected with the fluorenylmethoxycarbamate (FMOC) group. EDC (1.0 mmol) was added and the mixture was agitated over night. The mixture was filtered and then resubmitted to the same conditions for 4 hours.
  • DMAP dimethylaminopyridine
  • FMOC fluorenylmethoxycarbamate
  • Step B Deprotection of the N-Fmoc group.
  • the N-Fmoc protecting group was removed from the resin (75 mg , 0.015 mmol) from Step A by treatment with 20% piperidine in dimethylformamide for 30 minutes. Following filtration, the resin was washed sequentially with dimethylformamide (3 times), dichloromethane (1 time) and dimethylformamide (2 times) and used in the subsequent reaction.
  • Step C Coupling of the next N-Fmoc-amino acid derivative
  • a solution of the N-Fmoc-piperidine-4-carboxylic acid (0.06 mmol) in dimethylformamide (1 mL) was mixed with 2-(lH-benzotriazol- l-yl)-l,l,3,3-tetramethyluronium hexafluorophosphate (0.75 mmol), 1-N- hydroxybenztriazole (0.06 mmol) and diisopropylethylamine (0.07 mmol).
  • This solution was transferred to resin from Step B and typically allowed to react for 2 hours. Couplings were monitored by ninhydrin reaction. The coupling mixture was filtered and the resin washed with dimethylformamide (3 times) and used in the subsequent reaction.
  • Step D Deprotection of the N-Fmoc group.
  • the N-Fmoc protecting group was removed from the resin from Step C by the procedure described in Step B and used in the subsequent reaction.
  • Step E Acylation (or sulfonylation) of the terminal amino group.
  • the desired N-terminal capping reagent sulfonylchloride or acylchloride (0.06 mol) was dissolved in 50% CH 2 C1 2 /THF (2 ml), mixed with N,N-diisopropylethylamine(0.8 mmol) and added to the resin from Step D. After agitating over night, the resin was sequentially washed (3 x 3 ml 50% THF/CH 2 C1 2 ), 3 x 3 ml CH 2 C1 2 , 3 x 3 ml MeOH, 3 x 3 ml CH 2 C1 2
  • Step G Further modifications of product of Step E and Step F
  • the product of Step E and Step F may be forther modified by
  • Step F Cleavage of the desired products from the resins.
  • the final desired products were cleaved from the resins from Step E-H by mixing with a solution of 5% trifluoroacetic acid in CH 2 C1 2 (3 x 3 ml for 10 minutes each) and filtering. The filtrate was concentrated in vacuo. The residue was dissolved in 50% CH 3 CN/H 2 0 and lyophilised.
  • t-butyl tyrosine and t-butyl aspartate commercially available SASRIN resins was utilised and 1% trifluoroacetic acid in CH 2 C1 2 (10 x 1 ml for 3 minutes) was used for cleavage.
  • the eluant was run into 20% pyridine in methanol and the mixture was concentrated in vacuo. Purity was assesed by HPLC and molecular ions were obtained by electrospray ionization mass spectrometry to confirm the structure of each compound.
  • Step F Product of Step A; Note B:Step F with benzoic acid; Note C:Step F with phenylacetic acid; Note D: Step G with phenyl isocyanate; Note E: Step F with FMOC-4-aminophenyl acetic acid followed by Step D and Step F with benzoic acid; Note F: Step F with FMOC-4-aminophenyl acetic acid followed by Step D and Step F with phenylacetic acid; Note G: Step F with FMOC-4-aminophenyl acetic acid followed by Step D and Step F with 2-methylphenylacetic acid; Note H: Step F with FMOC-4- aminophenyl acetic acid followed by Step D and Step G with phenylisocyanate; Note I: Step F with FMOC-4-aminophenyl acetic acid followed by Step D and Step G with 2-methylphenylisocyanate; Note J: Step F with FMOC-4-aminophenyl acetic acid followed by Step D and Step G with 2-
  • reaction mixture was stirred for 2 hours, diluted with 50 ml of EtOAc and washed with IN HC1 (2 x 15 ml), saturated sodium bicarbonate solution (2 x 15 ml) and brine (1 x 15 ml).
  • the organic phase was dried over MgsO 4 , filtered and concentrated in vacuo. The residue was purified by flash chromatography over silica gel eluting with 50%EtOAc/hexanes to provide the product.
  • Step A The product of Step A (0.19 mmol) was treated with 10 ml of 1:1 CH 2 C1 2 :Et 2 NH and stirred at room temperature for 2 hours. The reaction mixture was concentrated in vacuo. The residue was then combined in 5 ml of DMF with 56.5 mg ( 0.23 mmol) of 4-(2-methyl- phenylureido)phenylacetic acid, 0.1 lg (0.29 mmol) of HBTU, 0.04 g (0.29 mmol) N-hydroxybenzotriazole hydrate and lastly, 0.1 ml (0.57 mmol) of diisopropylethylamine.
  • reaction mixture was stirred at room temperature over night, diluted with 50 ml of EtOAc and washed with IN HC1 (2 x 15 ml), saturated sodium bicarbonate solution (2 x 15 ml) and brine (1 x 15 ml).
  • the organic phase was dried over MgsO 4 , filtered and concentrated in vacuo.
  • the residue was purified by flash chromatography over silica gel eluting with 50%EtOAc/hexanes to provide the intermediate t-butyl ester.
  • the mixture was filtered and concentrated in vacuo.
  • the residue was dissolved in 5 ml of methanol and was treated with excess 2N NaOH solution.
  • the solution was stirred over night at room temperarature, concentrated in vacuo to remove methanol and was extracted with ether.
  • the aqueous phase was acidified with IN HCl solution and was extracted with ethyl acetate to provide, after drying over MgSO 4 , 48.8 mg of the desired product.
  • Step C Preparation of the title compound 47.2 mg (0.13 mmol) of the product of Step A was combined with 24 mg (0.13 mmol) of the product of Step B, 74 mg (0.2 mmol) of HBTU, 26 mg (0.2 mmol) of HOBt and 50 mg (0.4 mmol) of diisopropyl ethyl amine in 2 ml of methylene chloride. The reaction mixture was stirred over night at room temperature.
  • the solution was diluted with ethyl acetate (10 ml) and was washed with IN HCl (2 x 3 ml), saturated NaHCO 3 (2 x 5 ml), water (1 x 5 ml) and brine (1 x 5 ml) was dried over MgS0 4 .
  • the mixture was filtered and concentrated in vacuo and the residue was purified by preparatory thin layer chromatography over silica gel.
  • the product was treated with 3 ml of 1:1 TFA/CH 2 C1 2 for 4 hours and concentrated in vacuo to provide the title compound.
  • Step A Preparation of CS-1 Coated Plates.
  • Untreated 96 well polystyrene flat bottom plates were coated with bovine serum albumin (BSA; 20 ⁇ g/ml) for 2 hours at room temperature and washed twice with phosphate buffered saline (PBS).
  • BSA bovine serum albumin
  • PBS phosphate buffered saline
  • the albumin coating was next derivatized with 10 ⁇ g/ml 3-(2- pyridyldithio) propionic acid N-hydroxysuccinimide ester (SPDP), a heterobifunctional crosslinker, for 30 minutes at room temperature and washed twice with PBS.
  • SPDP 3-(2- pyridyldithio) propionic acid N-hydroxysuccinimide ester
  • the CS-1 peptide (Cys-Leu-His-Gly-Pro-Glu-Ile- Leu-Asp-Val-Pro-Ser-Thr), which was synthesized by conventional solid phase chemistry and purified by reverse phase HPLC, was next added to the derivatized BSA at a concentration of 2.5 ⁇ g/ml and allowed to react for 2 hours at room temperature. The plates were washed twice with PBS and stored at 4°C.
  • Step B Preparation of Fluor escentlv Labeled Jurkat Cells.
  • Jurkat cells obtained from the American Type Culture Collection (Rockville, MD; cat # ATCC TIB-152) were grown and maintained in RPMI-1640 culture medium containing 10% fetal calf serum (FCS), 50 units/ml penicillin, 50 ⁇ g/ml streptomycin and 2 mM glutamine. Fluorescence activated cell sorter analysis with specific monoclonal antibodies confirmed that the cells expressed both the ⁇ 4 and ⁇ l chains of VLA-4. The cells were centrifuged at 400xg for five minutes and washed twice with PBS.
  • FCS fetal calf serum
  • the cells were incubated at a concentration of 2 x 10 cells/ml in PBS containing a 1 ⁇ M concentration of a fluorogenic esterase substrate (2', 7'-bis-(2-carboxyethyl)-5-(and -6)- carboxyfluorescein, acetoxymethyl ester; BCECF-AM; Molecular Probes Inc., Eugene, Oregon; catalog #B-1150) for 30-60 minutes at 37°C in a 5% C ⁇ 2 air incubator.
  • the fluorescently labeled Jurkat cells were washed two times in PBS and resuspended in RPMI containing 0.25% BSA at a _ * final concentration of 2.0 x 10 cells/ml.
  • StepC Assay Procedure.
  • Compounds of this invention were prepared in DMSO at lOOx the desired final assay concentration. Final concentrations were selected from a range between 0.001 nM-100 ⁇ M.
  • Three ⁇ L of diluted compound, or vehicle alone, were premixed with 300 ⁇ L of cell suspension in 96-well polystyrene plates with round bottom wells. 100 ⁇ L aliquots of the cell /compound mixture were then transferred in duplicate to CS-1 coated wells. The cells were next incubated for 30 minutes at room temperature. The non-adherent cells were removed by two gentle washings with PBS.
  • the signal peptide as well as domains 1 and 2 of human VCAM were amplified by PCR using the human VCAM cDNA (R & D Systems) as template and the following primer sequences: 3'-PCR primer:5'-AATTATAATTTGATCAACTTAC CTGTCAATTCTTTTACAGCCTGCC-3'; 5'-PCR primer:
  • the 5'-PCR primer contained EcoRI and PvuII restriction sites followed by a Kozak consensus sequence (CCACC) proximal to the initiator methionine ATG.
  • the 3'-PCR primer contained a Bell site and a splice donor sequence. PCR was performed for 30 cycles using the following parameters: 1 min. at 94 C, 2 min. at 55 C, and 2 min. at 72 C.
  • the resulting PCR product of 650 bp was digested with EcoRI and Bell and ligated to expression vector pig-Tail (R & D Systems, Minneapolis, MN) digested with EcoRI and BamHL
  • the pig-Tail vector contains the genomic fragment which encodes the hinge region, CH2 and CH3 of human IgGl (GenBank Accession no. Z17370).
  • the DNA sequence of the resulting VCAM fragment was verified using Sequenase (US Biochemical, Cleveland, OH).
  • the fragment encoding the entire VCAM-Ig fusion was subsequently excised from pig-Tail with EcoRI and NotI and ligated to pCI-neo (Promega, Madison, WI) digested with EcoRI and NotI.
  • the resulting vector designated pCI-neo/VCAM-Ig was transfected into CHO-K1 (ATCC CCL 61) cells using calcium-phosphate DNA precipitation (Specialty Media, Lavalette, NJ).
  • Stable VCAM-Ig producing clones were selected according to standard protocols using 0.2-0.8 mg/ml active G418 (Gibco, Grand Island, NY), expanded, and cell supematants were screened for their ability to mediate Jurkat adhesion to wells previously coated with 1.5 ⁇ g/ml (total protein) goat anti-human IgG (Sigma, St. Louis, MO).
  • VCAM- Ig was purified from crude culture supematants by affinity chromatography on Protein A/G Sepharose (Pierce, Rockford, IL) according to the manufacturer's instructions and desalted into 50 mM sodium phosphate buffer, pH 7.6, by ultrafiltration on a YM-30 membrane (Amicon, Beverly, MA).
  • the labeled protein was separated from unincorporated isotope by means of a calibrated HPLC gel filtration column (G2000SW;
  • Step C VCAM-Ig Binding Assay.
  • Compounds of this invention were prepared in DMSO at lOOx the desired final assay concentration. Final concentrations were selected from a range between 0.001 nM-100 ⁇ M.
  • Jurkat cells were centrifuged at 400xg for five minutes and resuspended in binding buffer (25 mM HEPES, 150 mM NaCl, 3 mM KC1, 2 mM glucose, 0.1% bovine serum albumin, pH 7.4). The cells were centrifuged again and resuspended in binding buffer supplemented with MnCl 2 at a final concentration of 1 mM.
  • Compounds were assayed in Millipore MHVB multiscreen plates (cat# MHVBN4550, Millipore Corp., MA) by making the following additions to duplicate wells: (i) 200 ⁇ L of binding buffer
  • I-VCAM-Ig in the absence of cells was usually less than 5% of that observed using cells in the presence of vehicle. Percent inhibition was then calculated for each test well and the IC 50 was determined from a ten point titration using a validated four parameter fit algorithm.
  • Step A A ⁇ L Cell line.
  • RPMI-8866 cells (a human B cell line ⁇ ; a gift from Prof. John Wilkins, University of Manitoba, Canada) were grown in RPMI/10% fetal calf serum/ 100 U penicillin/100 ⁇ g streptomycin/2 mM L-glutamine at 37°C, 5 % carbon dioxide. The cells were pelleted at 1000 rpm for 5 minutes and then washed twice and resuspended in binding buffer (25 mM Hepes, 150 mM NaCl , 0.1 % BSA, 3 mM KC1, 2 mM Glucose, pH 7.4).
  • Step B VCAM-Ig Binding Assay.
  • Compounds of this invention were prepared in DMSO at lOOx the desired final assay concentration. Final concentrations were selected from a range between 0.001 nM-100 ⁇ M. Compounds were assayed in Millipore MHVB multiscreen plates (Cat# MHVBN4550) by making the following sequential additions to duplicate wells: (i) 100 ⁇ l/well of binding buffer containing 1.5 mM MnCl 2 ; (ii) 10 ⁇ l/well 125 I- VCAM-Ig in binding buffer (final assay concentration ⁇ 500 pM); (iii) 1.5 ⁇ l/well test compound or DMSO alone; (iv) 38 ⁇ l/well RPMI-8866 cell suspension (1.25 x 10 6 cells/well).
  • the plates were incubated at room temperature for 45 minutes on a plate shaker at 200 rpm, filtered on a vacuum box, and washed on the same apparatus by the addition of 100 ⁇ L of binding buffer containing 1 mM MnCl 2 .
  • 100 ⁇ L of Microscint-20 (Packard cat# 6013621) was added to each well.
  • the plates were then sealed, placed on a shaker for 30 seconds, and counted on a Topcount microplate scintillation counter (Packard).
  • Control wells containing DMSO alone were used to determine the level of VCAM-Ig binding corresponding to 0% inhibition.
  • Wells in which cells were omitted were used to determine the level of binding corresponding to 100% inhibition. Percent inhibition was then calculated for each test well and the IC 50 was determined from a ten point titration using a validated four parameter fit algorithm.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

Les 4-pipéridine carboxamides 4-susbstitués de Formule I sont des antagonistes de VLA-4 and/ou de α4β7 et, à ce titre, conviennent pour l'inhibition ou la prévention de l'adhésion cellulaire ou des pathologies à médiation par adhésion cellulaire. Ces composés peuvent être préparés sous forme de compositions pharmaceutiques et conviennent pour le traitement de l'asthme, des allergies, des inflammations, de la sclérose en plaques ainsi que pour diverses affections inflammatoires et maladies auto-immunes.
PCT/US1998/024513 1997-11-18 1998-11-16 Derives de 4-piperidine carboxamides 4-substitues WO1999025685A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU14150/99A AU1415099A (en) 1997-11-18 1998-11-16 4-substituted-4-piperidine carboxamide derivatives

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US6591797P 1997-11-18 1997-11-18
US60/065,917 1997-11-18
GB9727214.0 1997-12-23
GBGB9727214.0A GB9727214D0 (en) 1997-12-23 1997-12-23 4-Substituted-4-piperdine carboxamide derivatives

Publications (1)

Publication Number Publication Date
WO1999025685A1 true WO1999025685A1 (fr) 1999-05-27

Family

ID=26312836

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1998/024513 WO1999025685A1 (fr) 1997-11-18 1998-11-16 Derives de 4-piperidine carboxamides 4-substitues

Country Status (2)

Country Link
AU (1) AU1415099A (fr)
WO (1) WO1999025685A1 (fr)

Cited By (15)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001047887A1 (fr) * 1999-12-24 2001-07-05 Bayer Aktiengesellschaft COMPOSES D'ACIDE AMINE β UTILISES COMME ANTAGONISTES DE L'INTEGRINE
WO2001051487A1 (fr) * 1999-12-28 2001-07-19 Pfizer Products Inc. Inhibiteurs non peptidyle de liaison cellulaire dependant de vla-4 utiles pour le traitement de maladies inflammatoires, auto-immunes et respiratoires
EP1156807A1 (fr) * 1998-12-18 2001-11-28 Du Pont Pharmaceuticals Company N-ureidoalkyl-piperidines utilisees en tant que modulateurs de l'activite des recepteurs des chimiokines
WO2004099143A1 (fr) * 2003-05-09 2004-11-18 Glaxo Group Limited Derives d'amine cycliques, procedes de preparation de ceux-ci et compositions pharmaceutiques les contenant
WO2005016883A2 (fr) * 2003-08-14 2005-02-24 Icos Corporation Derives d'acrylamide servant d'antagonistes de l'integrine vla-1, et leurs utilisations
WO2005121135A1 (fr) * 2004-06-14 2005-12-22 Daiichi Pharmaceutical Co., Ltd. Inhibiteur de vla-4
US7053089B2 (en) 2001-02-23 2006-05-30 Merck & Co., Inc. N-substituted nonaryl-heterocyclic NMDA/NR2B antagonists
US7081460B2 (en) 2001-04-09 2006-07-25 Ortho-Mcneil Pharmaceutical, Inc. Quinazoline and quinazoline-like compounds for the treatment of integrin-mediated disorders
US7534893B2 (en) 2004-06-09 2009-05-19 Glaxo Group Limited Imidazol-2-one compounds useful in the treatment of various disorders
WO2009126920A2 (fr) 2008-04-11 2009-10-15 Merrimack Pharmaceuticals, Inc. Lieurs d'albumine de sérum humain, et ses conjugués
US7691843B2 (en) 2002-07-11 2010-04-06 Pfizer Inc. N-hydroxyamide derivatives possessing antibacterial activity
US7718673B2 (en) 1999-09-29 2010-05-18 Ortho-Mcneil Pharmaceutical, Inc. Isonipecotamides for the treatment of integrin-mediated disorders
EP2228065A3 (fr) * 2002-04-19 2010-12-22 Bristol-Myers Squibb Company Inhibiteurs heterocyclo de la fonction du canal potassique
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
CN104356051A (zh) * 2014-11-24 2015-02-18 苏州乔纳森新材料科技有限公司 一种镇痛药物中间体及其合成方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989010752A1 (fr) * 1988-05-13 1989-11-16 Abbott Laboratories Inhibiteurs de protease retrovirale
WO1995008536A1 (fr) * 1993-09-22 1995-03-30 Fujisawa Pharmaceutical Co., Ltd. Derives de n-(3-piperidinylcarbonyl)-beta-alanine utilises comme antagonistes du facteur d'agregation plaquettaire
WO1997003094A1 (fr) * 1995-07-11 1997-01-30 Biogen, Inc. Composes inhibiteurs d'adherence cellulaire
US5770575A (en) * 1994-03-16 1998-06-23 Ortho Pharmaceutical Corporation Nipecotic acid derivatives as antithrombotic compounds

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1989010752A1 (fr) * 1988-05-13 1989-11-16 Abbott Laboratories Inhibiteurs de protease retrovirale
WO1995008536A1 (fr) * 1993-09-22 1995-03-30 Fujisawa Pharmaceutical Co., Ltd. Derives de n-(3-piperidinylcarbonyl)-beta-alanine utilises comme antagonistes du facteur d'agregation plaquettaire
US5770575A (en) * 1994-03-16 1998-06-23 Ortho Pharmaceutical Corporation Nipecotic acid derivatives as antithrombotic compounds
WO1997003094A1 (fr) * 1995-07-11 1997-01-30 Biogen, Inc. Composes inhibiteurs d'adherence cellulaire

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, No. 114:247788, Vol. 114, 1991, RN 129739-41-9p; & WO 8910752 A1 (ABBOTT LABORATORIES) 16 November 1989. *
CHEMICAL ABSTRACTS, No. 123:285788; & WO 9508536 A1 (FUJISAWA PHARMACEUTICAL CO., LTD.) 30 March 1995. *
CHEMICAL ABSTRACTS, No. 126:47110, RN 184715-74, RN 184715-75; & JP 08277285 A (OOKUBO M. et al.) 22 October 1996. *

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1156807A4 (fr) * 1998-12-18 2002-04-03 Du Pont Pharm Co N-ureidoalkyl-piperidines utilisees en tant que modulateurs de l'activite des recepteurs des chimiokines
EP1156807A1 (fr) * 1998-12-18 2001-11-28 Du Pont Pharmaceuticals Company N-ureidoalkyl-piperidines utilisees en tant que modulateurs de l'activite des recepteurs des chimiokines
US7718673B2 (en) 1999-09-29 2010-05-18 Ortho-Mcneil Pharmaceutical, Inc. Isonipecotamides for the treatment of integrin-mediated disorders
US6794386B2 (en) 1999-12-24 2004-09-21 Bayer Aktiengesellschaft β-amino acid compounds as integrin antagonists
WO2001047887A1 (fr) * 1999-12-24 2001-07-05 Bayer Aktiengesellschaft COMPOSES D'ACIDE AMINE β UTILISES COMME ANTAGONISTES DE L'INTEGRINE
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
WO2001051487A1 (fr) * 1999-12-28 2001-07-19 Pfizer Products Inc. Inhibiteurs non peptidyle de liaison cellulaire dependant de vla-4 utiles pour le traitement de maladies inflammatoires, auto-immunes et respiratoires
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
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
US7053089B2 (en) 2001-02-23 2006-05-30 Merck & Co., Inc. N-substituted nonaryl-heterocyclic NMDA/NR2B antagonists
US7081460B2 (en) 2001-04-09 2006-07-25 Ortho-Mcneil Pharmaceutical, Inc. Quinazoline and quinazoline-like compounds for the treatment of integrin-mediated disorders
EP2228065A3 (fr) * 2002-04-19 2010-12-22 Bristol-Myers Squibb Company Inhibiteurs heterocyclo de la fonction du canal potassique
US7691843B2 (en) 2002-07-11 2010-04-06 Pfizer Inc. N-hydroxyamide derivatives possessing antibacterial activity
JP2006525975A (ja) * 2003-05-09 2006-11-16 グラクソ　グループ　リミテッド 環状アミン誘導体、その製法、およびそれを含有する医薬組成物
WO2004099143A1 (fr) * 2003-05-09 2004-11-18 Glaxo Group Limited Derives d'amine cycliques, procedes de preparation de ceux-ci et compositions pharmaceutiques les contenant
JP4684221B2 (ja) * 2003-05-09 2011-05-18 グラクソ グループ リミテッド 環状アミン誘導体、その製法、およびそれを含有する医薬組成物
WO2005016883A3 (fr) * 2003-08-14 2005-04-14 Icos Corp Derives d'acrylamide servant d'antagonistes de l'integrine vla-1, et leurs utilisations
WO2005016883A2 (fr) * 2003-08-14 2005-02-24 Icos Corporation Derives d'acrylamide servant d'antagonistes de l'integrine vla-1, et leurs utilisations
US7534893B2 (en) 2004-06-09 2009-05-19 Glaxo Group Limited Imidazol-2-one compounds useful in the treatment of various disorders
WO2005121135A1 (fr) * 2004-06-14 2005-12-22 Daiichi Pharmaceutical Co., Ltd. Inhibiteur de vla-4
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
CN104356051A (zh) * 2014-11-24 2015-02-18 苏州乔纳森新材料科技有限公司 一种镇痛药物中间体及其合成方法

Also Published As

Publication number Publication date
AU1415099A (en) 1999-06-07

Similar Documents

Publication Publication Date Title
EP1034164B1 (fr) Derives d'alanine-beta agissant en tant qu'inhibiteurs de l'adhesion cellulaire
AU728435B2 (en) Sulfonamides as cell adhesion inhibitors
AU726585B2 (en) Biarylalkanoic acids as cell adhesion inhibitors
US6353099B1 (en) Substituted ureas as cell adhesion inhibitors
US6645939B1 (en) Substituted β-alanine derivatives as cell adhesion inhibitors
US6403584B1 (en) Substituted nipecotyl derivatives as inhibitors of cell adhesion
WO1999020272A1 (fr) Acides azapeptidiques utilises comme inhibiteurs de l'adhesion cellulaire
US6271252B1 (en) Cyclic amino acid derivatives as cell adhesion inhibitors
WO1999026923A1 (fr) Derives de carboxamide de para-aminomethylaryl
US6191171B1 (en) Para-aminomethylaryl carboxamide derivatives
US6291511B1 (en) Biarylalkanoic acids as cell adhesion inhibitors
US6020347A (en) 4-substituted-4-piperidine carboxamide derivatives
US6221888B1 (en) Sulfonamides as cell adhesion inhibitors
EP1001764A1 (fr) Composes d'amide heterocycliques utilises en tant qu'inhibiteurs de l'adhesion cellulaire
WO1999064395A1 (fr) Composes d'amide heterocycliques utilises comme inhibiteurs d'adhesion cellulaire
US6579889B2 (en) Substituted isonipecotyl derivatives as inhibitors of cell adhesion
WO1999026922A1 (fr) Derives de pyrrole a substitution tenant lieu d'inhibiteurs d'adhesion cellulaire
WO1999025685A1 (fr) Derives de 4-piperidine carboxamides 4-substitues
US6559174B2 (en) N-arylsulfonyl aryl aza-bicyclic derivatives as potent cell adhesion inhibitors
GB2354440A (en) Aryl amides as cell adhesion inhibitors
CA2603835A1 (fr) Antagonistes du vla-4
US6482840B2 (en) Substituted cyclic amidine derivatives as inhibitors of cell adhesion
US6069163A (en) Azapeptide acids as cell adhesion inhibitors
US6855708B2 (en) N-arylsulfonyl aza-bicyclic derivatives as potent cell adhesion inhibitors
US6734311B2 (en) Substituted amidine derivatives as inhibitors of cell adhesion

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AL AM AU AZ BA BB BG BR BY CA CN CU CZ EE GD GE HR HU ID IL IS JP KG KR KZ LC LK LR LT LV MD MG MK MN MX NO NZ PL RO RU SG SI SK SL TJ TM TR TT UA US UZ VN YU

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW SD SZ UG ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE BF BJ CF CG CI CM GA GN GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: CA

点击 这是indexloc提供的php浏览器服务，不要输入任何密码和下载