JP2007023029A - Pharmaceutical composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pharmaceutical composition having α<SB>4</SB>-integrin-mediated cell adhesion inhibitory activity. <P>SOLUTION: The pharmaceutical composition contains as active ingredient a new phenylalanine compound represented by the formula(I) and having α<SB>4</SB>-integrin-mediated cell adhesion inhibitory activity or a pharmacologically acceptable derivative thereof. <P>COPYRIGHT: (C)2007,JPO&INPIT

Description

The present invention relates to a pharmaceutical composition comprising, as an active ingredient, an inhibitor of alpha ₄ integrin mediated cell adhesion.

  Multiple adhesion interactions between leukocytes and endothelial cells or extracellular matrix proteins are important factors in the regulation of immunity and inflammation. The first event in leukocyte migration from the blood vessel at the site of inflammation is rolling of leukocytes, followed by tight adhesion after integrin affinity changes (see Non-Patent Documents 1-7). In response to chemotactic factors, leukocytes pass through two adjacent endothelial cells, some of which are extracellular matrix proteins fibronectin (FN) (see Non-Patent Document 8) and collagen (CN) (non-patent It shifts to the organization which consists of literature 9 and nonpatent literature 10). These important recognition molecules involved in the adhesion reaction belong to the integrin gene superfamily (see Non-Patent Document 3, Non-Patent Document 11, Non-Patent Document 5, and Non-Patent Document 6).

  Integrins are heterodimers composed of non-covalently assembled subunits called alpha (α) and beta (β) subunits. To date, 8 integrin β subunits have been identified, which can bind to 16 different α subunits to form at least 23 different integrins.

Α ₄ β ₁ integrin, also known as VLA-4 (Very Late Antigen-4), is constitutively expressed on the surface of leukocytes such as lymphocytes, monocytes, eosinophils and basophils ( Non-patent document 12 and Non-patent document 13). It has been reported that VLA-4 is present in neutrophils of septic patients (see Non-Patent Document 14). VLA-4 binds to vascular cell adhesion molecule-1 (VCAM-1) on activated endothelial cells, and then leukocytes extravasate (see Non-Patent Document 15). When cells reach the extravascular space, VLA-4 binds to connecting segment 1 (CS-1), which is an alternative splicing region of the FNA chain (see Non-Patent Document 16). VLA-4 is also known to bind to osteopontin, a protein that is upregulated in atherosclerotic plaques (see Non-Patent Document 17).

Α ₄ β ₇ integrin, also known as LPAM-1 (Lymphocyte-Peyer's patch Adhesion Molecule-1), interacts with three known ligands (VCAM-1, CS-1, MAdCAM-1). A ligand having a specific specificity for α ₄ β ₇ is Mucosal Addressin Cell Adhesion Molecule-1 (MAdCAM-1) (Non-patent Document 18, Non-patent Document 19 and Non-patent Document). Reference 20). MAdCAM-1 is expressed at high levels in Peyer's patch high vascular endothelial venules, mesenteric lymph nodes, intestinal mucosa lamina propria and mammary veins (Non-patent Document 21). Integrins α ₄ β ₇ and MAdCAM-1 have been shown to be important in the regulation of lymphocyte transport to the normal small intestine (Non-patent Document 22).

In addition, α ₄ integrin α ₄ β ₁ and α ₄ β ₇ orally bioavailable non-peptide small molecule antagonists are asthma, inflammatory bowel disease, rheumatoid arthritis, multiple sclerosis and other It may be useful for treatment or prevention of pathological conditions such as diseases (see Patent Document 1 and Patent Document 2).

  The compounds used in the present invention fall within the comprehensive range of Patent Document 1, but are not specifically described in the application.

International Publication No. WO99 / 36393 International Publication WO02 / 18320 Batcher, Cell, 67: 1033-1036 (1991) Harlan, Blood, 3: 513-525 (1985) Hemler, Annu. Rev. Immunol., 8: 365-400 (1990) Osborn, Cell, 62: 3-6 (1990) Shimizu et al., Immunol. Rev., 114: 109-143 (1990) Springer, Nature, 346: 425-434 (1990) Springer, Cell, 76: 301-314 (1994) Wayner, J. Cell Biol., 105: 1873-1884 (1987) Bornstein et al., Ann. Rev. Biochem., 49: 957-1003 (1980) Miller, Extracellular Matrix Biochemistry, “Collagen Chemistry and Its Distribution,” K.A. Piers and A.H. Regis, Elsevier Publishing, Amsterdam, 41-78 (1983) Hynes, Cell, 48: 549-554 (1987) Hemler et al., J. Bio. Chem. 262: 11478-11485 (1987) Bochner, J. Exp. Med. 173: 1553-1556 (1991) Ibbotson et al., Nature Med. 7: 465-470 (2001) Elices et al., Cell 60: 577-584 (1990) Wine et al., J. Cell Biol., 109: 1321-1330 (1989) Bayless et al., J. Cell Science 111: 1165-1174 (1998) Andrew et al., J. Immunol. 153: 3847-3861 (1994) Briskin et al., Nature 363: 461-464 (1993) Shyjan et al., J. Immunol. 156: 2851-2857 (1996) Berg et al., Immunol. Rev. 105: 5-18 (1989) Holzmann et al., Cell 56: 37-46 (1989)

The present invention aims to provide a pharmaceutical composition having alpha ₄ integrin mediated cell adhesion inhibitory activity.

（Ｉ）
（式中、Ｒ¹はブロモ、Ｒ²はハロゲン、Ｃ_1-6アルキルまたはＣ_1-6アルコキシを表す）
で示される化合物、またはその薬理学的に許容しうる誘導体を有効成分として含有することを特徴とする医薬組成物を提供する。 The present inventors have conducted extensive studies to solve the above problems, found a novel pharmaceutical composition comprising, as an active ingredient a phenylalanine derivative having an inhibitory effect on alpha ₄ integrin mediated cell adhesion, the present invention completed.
That is, the present invention relates to formula (I):

(I)
(Wherein R ¹ represents bromo, R ² represents halogen, C _1-6 alkyl or C _1-6 alkoxy)
Or a pharmacologically acceptable derivative thereof as an active ingredient.

For the present invention, compound (I) wherein R ² is halogen or C _1-6 alkoxy or a pharmaceutically acceptable derivative thereof is preferred.
Further, a compound in which R ² is fluoro, methoxy or ethoxy or a pharmacologically acceptable derivative thereof is more preferable.

Further preferred compounds for the present invention are P1 to P7 (described later) or pharmacologically acceptable derivatives thereof. Specifically,
(S) -2-{[1- (2-Bromo-5-ethoxyphenyl) methanoyl] amino} -3- (4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl) propionic acid,
(S) -2-{[1- (2-bromo-5-fluorophenyl) methanoyl] amino} -3- (4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl) propionic acid,
(S) -2-{[1- (2-bromo-5-methoxyphenyl) methanoyl] amino} -3- (4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl) propionic acid,
(S) -2-{[1- (2-bromo-5-methylphenyl) methanoyl] amino} -3- (4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl) propionic acid,
(S) -2-{[(2-bromo-5-chlorophenyl) methanoyl] amino} -3- [4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl] propionic acid,
(S) -2-{[(2,5-dibromophenyl) methanoyl] amino} -3- [4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl] propionic acid, and
(S) -2-{[(5- (iso-propoxy) -2-bromophenyl) methanoyl] amino} -3- [4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl] propionic acid Or a pharmacologically acceptable derivative thereof.

In the present specification, the terms described below have the following meanings unless otherwise specified.
“Halogen” means a group selected from fluorine, chlorine, bromine or iodine.

“C _1-6 alkyl” means a straight or branched alkyl group having 1 to 6 carbon atoms, or a part of a group containing them, such as methyl, ethyl, propyl, isopropyl, n- Examples include butyl, isobutyl, tert-butyl, pentyl and hexyl.

“C _1-6 alkoxy” means a group in which an oxygen atom is bonded to the above C _1-6 alkyl group or a part of a group containing them, such as methoxy, ethoxy, propoxy, isopropoxy, n- Examples include butoxy, isobutoxy, tert-butoxy or hexoxy.

  The compound according to the present invention has a 2,5-2 substituted benzoyl group and is characterized in that a cyano group is introduced at the 4′-position of the biphenyl nucleus.

Compound or the pharmaceutically acceptable derivatives of formula (I) have a potent inhibitory activity on alpha ₄ integrin mediated cell adhesion. Furthermore, it has been found that certain preparative compounds show excellent bioavailability and / or good systemic exposure after oral administration.
Compounds P1, P2 and P3 (described below) show an advantageous combination of the above characteristics.

  It will be appreciated that a compound of formula (I) or a pharmaceutically acceptable derivative thereof may have more than one asymmetric carbon atom, but in that case may exist as a diastereomer. All such isomeric forms and mixtures can also be used in the pharmaceutical composition of the present invention.

  Separation of diastereomers can be achieved by usual methods such as fractional crystallization, chromatography, or HPLC. Alternatively, single stereoisomeric compounds of the invention may be prepared, if necessary, from the optically pure corresponding intermediate, or the corresponding racemate is resolved using an appropriate chiral support (e.g. Or by fractional crystallization of diastereomeric salts formed by reaction of the corresponding racemate with the appropriate optically active acid or base. Alternatively, a mixture of enantiomers can be separated by chemical reaction with an appropriate chiral compound to form a new covalent species. For example, coupling of a racemic carboxylic acid with a chiral amine or chiral alcohol yields a diastereomeric mixture (amide or ester, respectively), which can be separated by conventional techniques such as column chromatography, HPLC or fractional crystallization. it can. The new covalent bond can then be cleaved with a suitable chemical reaction such as hydrolysis to convert one diastereomeric form into one enantiomer form of the desired compound.

  In the present specification, the “pharmacologically acceptable derivative” means a pharmacologically acceptable salt or prodrug (eg, ester) of the compound according to the present invention, and these are administered to a patient. The compounds according to the invention or their active metabolites or residues can be given (directly or indirectly). Such derivatives can be obtained by one skilled in the art without undue experimentation. See, for example, Burger's Medicinal Chemistry and Drug Discovery, 5th edition, Volume 1, “Principles and Practice”. Preferred pharmacologically acceptable derivatives are salts and esters.

  A person skilled in the field of organic chemistry knows that a large number of organic compounds can form complexes or precipitate or crystallize with the solvent of the reaction system. These complexes are widely known as “solvates”. For example, a complex with water is known as a “hydrate”. Solvates of the compounds according to the invention are also within the scope of the invention.

  In the present specification, the “prodrug” means a compound that is converted into an active form having a pharmacological action by hydrolysis in the body, for example, in blood. Examples of pharmacologically acceptable prodrugs can be found in the literature [T. Higuchi and V. Stella, Prodrugs as Novel Delivery Systems, “Bioreversible Carriers in Drug Design. Design) ", Edward B. Roche, Ed., American Pharmaceutical Association and Pergamon Press, ACS Symposium Series, Volume 14, (1987); and D. Fleischer, S. Ramon and H. Barbara , “Improved oral drug delivery: solubility limitations overcome by the use of prodrugs, Advanced Drug Delivery Reviews (1996) 19 (2): 115-130” ]It is described in.

  Prodrugs are carriers covalently linked to a compound or derivative that release a compound of formula (I) or a pharmacologically acceptable derivative in vivo when administered to a patient. Prodrugs are usually prepared by modifying functional groups such that the modifying moiety is cleaved by conventional means or in vivo to yield the parent compound. In the case of carboxylic acid (—COOH), esters such as methyl ester, ethyl ester and double ester can also be used. Esters are those that are active in humans and / or hydrolyzed under in vivo conditions, and suitable pharmacologically acceptable in vivo hydrolysable ester groups include those in the human body. Those which are easily decomposed to liberate the acid of the parent compound or a salt thereof.

  The compounds according to the invention may be in the form of pharmacologically acceptable salts and / or may be administered as pharmacologically acceptable salts. Suitable salts are reviewed in the literature (Berge et al., J. Pharm. Sci., 66: 1-19 (1977)).

  Usually, a pharmacologically acceptable salt can be easily produced by using a desired acid or base as required. The salt can be recovered by precipitating from solution and filtering or by distilling off the solvent.

  In the case of compounds of formula (I), suitable pharmacologically acceptable salts are formed from pharmacologically acceptable bases. Such base salts include ammonium salts, alkali metal salts such as sodium and potassium salts, alkaline earth metal salts such as calcium and magnesium, and primary, secondary and tertiary amines (eg, isopropylamine, diethylamine). And salts with organic bases, including salts of ethanolamine, trimethylamine, dicyclohexylamine, N-methyl-D-glucamine and tris (hydroxymethyl) methylamine).

(II)
（式中、Ｒ¹およびＲ²は式(Ｉ)における定義と同意義であり、Ｒはカルボン酸エステルを形成しうる基を表す）
で示されるカルボン酸エステル誘導体を加水分解し、適宜、次いで薬理学的に許容しうる誘導体を形成することにより製造することができる。 Compound (I), which is an active ingredient of the present invention, has the formula (II):

(II)
(Wherein R ¹ and R ² are as defined in formula (I), and R represents a group capable of forming a carboxylic acid ester)
Can be produced by hydrolyzing the carboxylic acid ester derivative represented by the formula, and then forming a pharmacologically acceptable derivative as appropriate.

R is preferably C _1-6 alkyl such as methyl or t-butyl, and more preferably methyl. Hydrolysis is performed using an acidic or alkaline solvent. An example of hydrolysis in an alkaline solvent is treatment of the compound of formula (II) with an alkali metal hydroxide in a suitable solvent, such as treatment with lithium hydroxide in an aqueous tetrahydrofuran solution. As an example of hydrolysis in an acidic medium, treatment of a compound of formula (II) with an inorganic acid at an elevated temperature in a suitable co-solvent, eg overnight treatment with 5N hydrochloric acid at 60 ° C. in dioxane Is mentioned. Those skilled in the art are familiar with such methods.

(III)
（式中、Ｒは式(II)における定義と同意義である）
で示される化合物またはその酸付加塩を式（IV）：

(IV)
（式中、Ｒ¹およびＲ²は式(Ｉ)における定義と同意義であり、Ｘは水酸基または脱離基である）
で示される化合物と反応させることにより製造することができる。 The compound of formula (II) is represented by formula (III):

(III)
(Wherein R has the same meaning as defined in formula (II))
Or a compound thereof represented by formula (IV):

(IV)
(Wherein R ¹ and R ² are as defined in formula (I), and X is a hydroxyl group or a leaving group)
It can manufacture by making it react with the compound shown by these.

  The acid addition salt of the compound of formula (III) is preferably a hydrochloride. When X is a leaving group, the leaving group X is preferably halogen, especially chloro. Usually, the reaction of the compound of formula (III) with the compound of formula (IV) is carried out in an inert organic solvent such as tetrahydrofuran or dichloromethane or in a mixed organic solvent / aqueous solvent, with an organic base (eg triethylamine), alkaline Performed at room temperature or elevated temperature in the presence of a suitable base such as a metal carbonate (eg, potassium carbonate) or an alkali metal bicarbonate (eg, sodium bicarbonate). When X is a hydroxyl group, the reaction of the compound of formula (III) with the compound of formula (IV) can be carried out using standard coupling methods (eg O- (7-azabenzotriazol-1-yl) -N, N, N ′, N′-tetramethyluronium hexafluorophosphate, triethylamine is stirred in dry dimethylformamide at room temperature overnight).

(V)
（式中、Ｚは脱離基であり、Ｒ^aは保護基である）
で示される化合物を式（VI）：

(VI)
（式中、Ｒ^bはヒドロキシメチル基である）
で示される化合物とカップリングさせ、次いで保護基Ｒ^aを除去し、Ｒ^bをシアノ基に変換することにより製造することができる。得られた生成物の保護基は、下記の方法により除去することができる。当業者が精通している方法及び本明細書記載の方法を用いてＲ^bをシアノ基に変換することができる。 The compound of formula (III) is represented by formula (V):

(V)
(Wherein Z is a leaving group and R ^a is a protecting group)
A compound represented by formula (VI):

(VI)
(Wherein R ^b is a hydroxymethyl group)
And then removing the protecting group R ^a and converting R ^b to a cyano group. The protecting group of the obtained product can be removed by the following method. R ^b can be converted to a cyano group using methods familiar to those skilled in the art and methods described herein.

  The reaction of the compound of formula (V) with the compound of formula (VI) can be carried out, for example, under Suzuki coupling conditions familiar to those skilled in the art. For example, a suitable base (eg, triethylamine) at elevated temperature in a suitable solvent (eg, 1-methyl-2-pyrrolidone) using a palladium catalyst (eg, tetrakis (triphenylphosphine) palladium (0)) The reaction can be carried out in the presence of The resulting protecting groups of the product can be removed by methods familiar to those skilled in the art. When the protecting group is t-butyloxycarbonyl, deprotection can be accomplished by acid treatment (eg treatment with hydrochloric acid) in a suitable solvent (eg alcohol such as ethanol or isopropanol).

  Intermediate compounds (IV) and (V) are commercially available or can be prepared by methods known to those skilled in the art using methods described herein or by methods analogous thereto. .

  One skilled in the art will recognize one or more reactive groups in the molecule to prevent undesired side reactions when preparing the compound of formula (I) or a pharmaceutically acceptable derivative thereof according to the present invention. It will be appreciated that it is necessary and / or desirable to protect Suitable protecting groups for the present invention are well known to those skilled in the art and can be used according to conventional methods. See, for example, T.W. Green and P.G.M. Utz, “Protective groups in organic synthesis”, John Wiley & Sons (1991) and P.J. Kohiensky, “Protecting groups” Georg Thieme Verlag (1994). Suitable amino protecting groups include, for example, acyl-type protecting groups (formyl, trifluoroacetyl, acetyl, etc.), aromatic urethane-type protecting groups (benzyloxycarbonyl (Cbz) and substituted Cbz, etc.), aliphatic urethane protecting groups ( 9-fluorenylmethoxycarbonyl (Fmoc), t-butyloxycarbonyl (Boc), isopropyloxycarbonyl, cyclohexyloxycarbonyl and the like) and alkyl-type protecting groups (benzyl, trityl, chlorotrityl and the like). Suitable oxygen protecting groups include, for example, alkylsilyl groups such as trimethylsilyl or tert-butyldimethylsilyl, alkyl ethers such as tetrahydropyranyl or tert-butyl, or esters such as acetate.

  The biological activity of the compound that is the active ingredient of the pharmaceutical composition of the present invention is determined by the interaction between VLA-4 expressed on the Jurkat J6 cell membrane and the test compound by the Jurkat J6 scintillation proximity assay (SPA). Based on this, it can be tested in vitro. The drug useful as an active ingredient of the pharmaceutical composition of the present invention has a pKi of preferably about 8.6 or more when tested by the method of Test Example 1. All of the compounds obtained in the production examples described later have a pKi of 8.6 or more, and were found to be useful as the pharmaceutical composition of the present invention.

Since the compounds of formula (I) or pharmaceutical compositions of the present invention containing a pharmacologically acceptable derivative inhibits alpha ₄ integrin mediated cell adhesion, the following which are alpha ₄ integrin mediated cell adhesion is involved It can be used for the treatment or prevention of pathological conditions. For example, rheumatoid arthritis (RA); asthma; allergic diseases such as rhinitis; adult respiratory distress syndrome; AIDS dementia; Alzheimer's disease; cardiovascular disease; thrombosis or harmful platelet aggregation; reocclusion after thrombolysis; Injuries; skin inflammatory diseases such as psoriasis, eczema, contact dermatitis and atopic dermatitis; diabetes (eg, insulin-dependent diabetes, autoimmune diabetes); multiple sclerosis; systemic lupus lupus erythematosus (SLE); Inflammatory bowel diseases such as ulcerative colitis, Crohn's disease (local enterocolitis) and cystitis (eg, bowel disease resulting after colorectal resection and ileal anastomosis); Diseases involving leukocyte infiltration into the gastrointestinal tract, such as enteric diseases, lymphocytic or collagenous colitis, and eosinophilic gastroenteritis, such as skin, urinary tract, airways and joint synovium Diseases related to leukocyte infiltration into the epithelial membrane tissue of the pancreas; pancreatitis; mastitis (mammary gland); hepatitis; cholecystitis; cholangitis or cholangitis (biliary and hepatic surrounding tissue); bronchitis; sinusitis; Inflammatory diseases of the lung that cause interstitial fibrosis such as pneumonia; collagen disease (SLE and RA); sarcoidosis; osteoporosis; osteoarthritis; atherosclerosis; neoplasms with metastasis or cancerous growth Disease; trauma (enhanced wound healing); certain eye diseases such as retinal detachment, allergic conjunctivitis and autoimmune uveitis; Sjogren's syndrome; rejection after organ transplantation (chronic and acute); host versus graft or transplant Unilateral host disease; intimal thickening; arteriosclerosis (including graft arteriosclerosis after transplantation); surgery such as percutaneous transluminal coronary angioplasty (PTCA) and percutaneous transluminal artery recanalization Later reinfarction or restenosis; nephritis; tumor neovascularization ; Malignancy; multiple myeloma and myeloma-induced bone resorption; sepsis; stroke, central nervous injuries, such as traumatic brain injury and spinal cord injury; Meniere's disease and inflammatory bowel disease.

  The pharmaceutical composition of the present invention has allergic pathologies such as asthma and rhinitis, inflammatory bowel diseases such as ulcerative colitis and Crohn's disease, rheumatoid arthritis, atopic dermatitis, multiple sclerosis and rejection after organ transplantation. It can be suitably used for treatment or prevention. In particular, the pharmaceutical composition of the present invention is useful for the treatment or prevention of asthma, rheumatoid arthritis, inflammatory bowel disease or multiple sclerosis.

The pharmaceutical compositions of the present invention comprise the active compound (I) in a safe and effective amount, inhibition of alpha ₄ integrin mediated cell adhesion is useful in the treatment or prevention of beneficial conditions. In particular, the pharmaceutical composition of the present invention is useful in the treatment or prevention of the above-mentioned pathological conditions.

  The pharmaceutical composition of the present invention contains the active compound represented by formula (I) in a pharmacologically effective amount together with a pharmaceutically acceptable carrier or diluent.

  The pharmaceutical composition of the present invention can be produced by mixing at least one compound (I) or a pharmaceutically acceptable derivative thereof together with a pharmaceutically acceptable carrier or diluent.

  The pharmaceutical compositions of the present invention are in the form of human and veterinary pharmaceutical formulations for use in humans or animals, and usually contain one or more pharmaceutically acceptable diluents, carriers or excipients. contains. Suitable carriers or diluents for therapeutic use are well known in the pharmaceutical art and are described, for example, in the literature (Remington's Pharmaceutical Sciences, Mack Publishing Co, edited by A. R. Genaro, (1985), etc.). The choice of pharmaceutically acceptable carrier, excipient or diluent should be selected in light of the intended route of administration and normal pharmaceutical practice. The carrier or diluent must be acceptable in the sense that it is not harmful to the patient. Pharmaceutically acceptable carriers or diluents include, for example, binders (syrup, gum arabic, gelatin, sorbit, tragacanth, polyvinylpyrrolidone, etc.), excipients (lactose, sugar, corn starch, potassium phosphate, sorbit) And glycine), lubricants (magnesium stearate, talc, polyethylene glycol, silica, etc.), disintegrants (potato starch, etc.) and wetting agents (sodium lauryl sulfate, etc.).

  Examples of the administration (delivery) route of the pharmaceutical composition of the present invention include, but are not limited to, one or more exemplified below. Oral (tablets, capsules or solutions for oral ingestion, etc.), local, transmucosal (nasal spray or aerosol for inhalation, etc.), nasal, parenteral (by injection, etc.), gastrointestinal tract, spinal cord, intraperitoneal, muscle Internal, intravenous, intrauterine, intraocular, intradermal, intracranial, intratracheal, intravaginal, intraventricular, intracerebral, subcutaneous, intraocular (including intravitreal or intraocular), transdermal, transrectal, Oral mucosa, epidural, sublingual.

  For example, the compound may be administered orally in the form of tablets, capsules, ovules, elixirs, solutions or suspensions suitable for immediate, delayed, modified, sustained, pulsed or controlled release. it can. These preparations may contain fragrances or colorants. Tablets include excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine, starch (preferably corn, potato or tapioca starch), sodium starch glycolate, croscarmellose sodium and It may contain disintegrants such as certain silicic acid complexes and granulating binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia. In addition, lubricants such as magnesium stearate, stearic acid, glyceryl behenate and talc may be included. Similar types of solid ingredients can also be utilized as fillers in gelatin capsules. Preferred excipients in this regard include lactose, starch, cellulose, lactose or high molecular weight polyethylene glycols. For aqueous suspensions and / or elixirs, the drug can be any of various sweetening or flavoring agents, coloring materials or pigments, emulsifiers and / or suspending agents, and diluents such as water, ethanol, propylene glycol and glycerin. Or a combination thereof.

  In formulation, the compound of the present invention can be pulverized to a particle size suitable for tablet molding and other formulation types using a known pulverization method such as wet pulverization. Finely divided (nanoparticle) preparations of the compounds of the invention can be produced by methods known in the art (see eg WO 02/00196).

  When the pharmaceutical composition of the present invention is administered parenterally, for example, via an administration route such as intravenous, intraarterial, intraperitoneal, intrathecal, intraventricular, intraurinary, intrasternal, intracranial, intramuscular or subcutaneous. And / or by infusion. For parenteral administration, the drug is best used in the form of a sterile aqueous solution, which contains sufficient salt or glucose to produce other substances, for example, isotonic solutions with blood. Also good. The aqueous solution is appropriately buffered if necessary (pH 3-9 is preferred). The production of parenteral preparations under sterile conditions can be easily performed by conventional pharmaceutical techniques well known to those skilled in the art.

  As noted above, the pharmaceutical compositions of the present invention can be administered intranasally or by inhalation, including from pressurized containers, pumps, sprays or nebulizers, dichlorodifluoromethane, tolchlorofluoromethane, dichlorotetrafluoroethane, Hydrofluoroalkanes such as 1,1,1,2-tetrafluoroethane (HFA 134AT "") or 1,1,1,2,3,3,3-heptafluoropropane (HFA 227EA) (eg Ineos Fluor) Delivery in the form of dry powder inhalation or aerosol spray using a suitable propellant such as carbon dioxide or other suitable gas is convenient. In the case of a pressurized aerosol, the dosage unit can be determined by providing a valve to deliver a measured amount. Pressurized containers, pumps, sprays or nebulizers contain, for example, drug solutions or suspensions with a mixture of ethanol and propellant as a solvent, which further includes lubricants such as sorbitan trioleate May be contained. Cartridges (eg, gelatinous) for use in capsules and inhalers can be formulated containing a powder mixture of drug and a suitable powder base (such as lactose or starch).

  Alternatively, the pharmaceutical composition of the present invention may be a suppository or vaginal suppository dosage form, or a dosage form for topical application of a gel, hydrogel, lotion, solution, cream, ointment or spray. . In addition, the pharmaceutical composition of the present invention can be made into a transdermal dosage form such as a skin patch. Furthermore, the dosage form may be administered via the pulmonary or rectal route. Moreover, it can be set as the dosage form for intraocular administration. For intraocular application, the drug may be formulated as a fine powder suspension in sterile saline, pH adjusted to isotonicity, or preferably with a preservative such as benzylalkonium chloride. It can be formulated as a solution in sterile physiological saline adjusted to be isotonic pH. Alternatively, it can be formulated in an ointment such as petrolatum.

  The agents of the present invention for topical application to the skin are formulated, for example, as a suitable ointment containing a drug suspended or dissolved in a mixture of one or more of the following ingredients. Mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene polyoxypropylene compound, emulsifying wax and water. Alternatively, it can be formulated as a suitable lotion or cream suspended or dissolved in one or a mixture of two or more of the following ingredients. Mineral oil, sorbitan monostearate, polyethylene glycol, liquid paraffin, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

The pharmaceutical composition of the present invention can be administered by direct injection.
The pharmaceutical composition of the present invention is preferably administered systemically (orally, buccal, sublingual, etc.), more preferably oral administration. Accordingly, the drug of the present invention is preferably in a dosage form suitable for oral administration.

Usually, the optimal dose for an individual patient is determined by a physician. The specific dose level and frequency of administration for a particular individual will depend on the activity of the drug used, metabolic stability and duration of action, individual age, weight, overall health, gender and diet, mode of administration and duration, excretion It can vary depending on a variety of factors, including rate, concomitant medication, severity of the particular condition, and even the treatment the individual is receiving.
For oral and parenteral administration to humans, the daily dose of the drug may be a single dose or a divided dose.

  The dose of the compound of the present invention to a human (body weight of about 70 kg) is 0.1 mg to 2 g, preferably 0.1 mg to 1 g (as the weight of free acid), as the active ingredient per unit dose. The unit dose may be administered, for example, 1 to 4 times per day. The dose depends on the route of administration. Depending on the age and weight of the patient and the severity of the condition being treated, dosages will need to be routinely changed. The actual dose and route of administration will ultimately be at the discretion of the attending physician or veterinarian.

  The compounds according to the invention may also be used in combination with other therapeutic agents. Accordingly, the present invention provides a combination preparation comprising a compound according to the present invention or a pharmacologically acceptable derivative thereof together with other therapeutic agents.

  When a compound of formula (I) or a pharmacologically acceptable derivative thereof is used in combination with a second drug that is active against the same disease pathology, the dose of each drug is when they are used alone The dose may be different. Appropriate doses can be readily appreciated by one skilled in the art. The amount of drug according to the present invention required for use in therapy or prevention will vary depending on the nature of the condition being treated and the age and symptoms of the patient and will ultimately be determined by the attending physician or veterinarian. It is entrusted to. Other active drugs that can be combined with compounds of formula (I) or pharmacologically acceptable derivatives thereof include, but are not limited to, those exemplified below. (a) other VLA-4 antagonists, (b) H1 histamine antagonists, (c) NSAIDs, (d) anti-diabetic drugs such as glitazones, (e) anticholinergics, (f) 2- (4-ethoxy COX-2 inhibitors such as -phenyl) -3- (4-methanesulfonyl-phenyl) pyrazolo [1,5-b] pyridazine (disclosed in WO99 / 12930), (g) PDE-IV inhibitors, (h) Steroids such as corticosteroids, (i) β agonists, (j) chemokine receptor antagonists such as CCR-2, CCR-3, CCR-5 and CCR-8, (k) appropriate multiples such as interferon Sclerosis therapeutic or prophylactic agent, (l) LFA-1 antagonist, (m) TNF inhibitor, (n) sulfasalazine and 5-amino salicylate, and (o) immunosuppressant .

  Pharmaceutical formulations for using the above combinations can be provided in a conventional manner, and therefore the present invention also includes pharmaceutical formulations containing the above combinations together with a pharmaceutically acceptable carrier or excipient. To do. The individual components in such combinations can be administered sequentially or simultaneously from any convenient route of administration, as separate pharmaceutical formulations or combined pharmaceutical formulations. In the case of continuous administration, either the drug according to the present invention or the second therapeutic drug is administered first. In the case of simultaneous administration, the drugs used in combination can be administered in the same pharmaceutical preparation or in different pharmaceutical preparations.

  When used together in the same pharmaceutical formulation, the two drugs must be stable and compatible (coexisting) with each other and with other formulation components in the formulation. When formulated separately, each drug can be formulated in any convenient manner, but it is convenient to formulate each drug in a known manner.

The production method and the like of the compound which is an active ingredient of the present invention will be described below with reference to test examples, reference examples and production examples.
Test Example 1: Jarkat J6 Scintillation Proximity Assay (SPA)
The interaction between integrin VLA-4 (very late antigen-4; α4β1; CD49d, CD29) expressed on Jurkat J6 cell membrane and the test compound was examined by Jurkat J6 scintillation proximity assay.
Method
Wheat germ agglutinin coated SPA beads (Amersham, 1 mg / well) in J6 cells (1 million cells / well) in assay buffer containing 50 mM HEPES, 100 mM NaCl and 1 mM MnCl ₂ (pH adjusted to 7.5 with 4 M NaOH) ). Tritiated ³ H standard compound A (final concentration in assay 1 to ³ nM) and test compound are dissolved in a suitable solvent and diluted with assay buffer. Compounds are assayed once and fit to the four parameter curve fit of equation (I). The equilibrium dissociation constant of each compound was calculated according to the method of Cheng & Prusoff. Results (in vitro α ₄ β ₁ activity) were expressed in pKi.

（式中、ａは最小値、ｂは傾き、cはIC₅₀、ｄは最大値である。）
ここで、最大値および最小値は、化合物の非存在下、および2mM EDTA二カリウム塩(シグマケミカルズ,UK、製品番号ED2P)の存在下での接着と比較した場合の値である。データは平均pIC₅₀で示す。
後述の製造例で得た化合物を上記の方法で試験したところ、以下の表に示すように、全ての場合、pＫiは８.６以上であった。
表 １ ジャルカットJ6シンチレーション近接アッセイ Standard compound A is (2S) -3- [4-({[4- (aminocarbonyl) -1-piperidyl] carbonyl} oxy) phenyl] -2-[((2S) -4-methyl-2-{[ 2- (2-methylphenoxy) acetyl] amino} pentanoyl) amino] propanoic acid potassium salt (WO 00/37444). Production of the tritiated ³ H derivative was carried out according to a conventional method.
Equation (I):

(In the formula, a is the minimum value, b is the slope, c is IC ₅₀ , and d is the maximum value.)
Here, the maximum and minimum values are values compared to adhesion in the absence of compound and in the presence of 2 mM EDTA dipotassium salt (Sigma Chemicals, UK, product number ED2P). Data are shown as mean pIC ₅₀ .
When the compounds obtained in the production examples described later were tested by the above method, the pKi was 8.6 or more in all cases, as shown in the following table.
Table 1. Jurkat J6 Scintillation Proximity Assay

Unless otherwise stated in the general method experiment section, the term MDAP means Mass Directed Auto Preparation. This is an automated system for compound purification by preparative HPLC that uses a mass spectrometer in combination with a preparative HPLC system to detect and recover at the desired mass. The following employs a Waters Fraction Lynx MDAP system (water / acetonitrile gradient, both containing 0.1% formic acid) using a suitable reverse phase column.

Reference Example 1: (S) -2-tert-butoxycarbonylamino-3- (4-hydroxyphenyl) propionic acid methyl ester (S1)
Di-tert-butyl dicarbonate (200 g, 0.92 mol) was added to L-tyrosine methyl ester hydrochloride (200 g, 0.86 mol, Aldrich) and sodium hydrogen carbonate (100 g, 1.19 mol) in dichloromethane (1 L) and To the mixture in water (1 L) was added in small portions. The mixture was stirred at room temperature for 2 hours. The organic layer was separated and the aqueous phase was extracted again with dichloromethane (500 mL). The combined organic layers were dried over magnesium sulfate, filtered and concentrated under reduced pressure to give the crude title compound as a colorless glass. This was used in the next step without purification. LC / MS (ES-ve): [MH] ⁻ ; m / z 294 (C ₁₅ H ₂₁ NO ₅ ; [MH] ⁻ ; m / z 294).

Reference Example 2: (S) -2-tert-butoxycarbonylamino-3- (4-trifluoromethanesulfonyloxyphenyl) propionic acid methyl ester (S2)
Pyridine (58 mL, 0.72 mol) was added to crude dichloromethane (S) -2-tert-butoxycarbonylamino-3- (4-hydroxyphenyl) propionic acid methyl ester (S1, 70.5 g, 0.24 mol) under argon. 1 L) was added to the solution. The solution was ice-cooled, and trifluoromethanesulfonic anhydride (52 mL, 0.31 mol) was added dropwise with stirring. After the addition was complete, the mixture was stirred in ice for 2 hours, washed with 2M hydrochloric acid (500 mL), dried over magnesium sulfate, filtered and concentrated in vacuo. The crude product was purified by silica gel chromatography (Biotage 75L, silica 800 g, elution solvent; ethyl acetate: hexane (15:85)) to give the title compound as a colorless oil. The product slowly solidified to give a white solid; LC / MS (ES + ve): [M-BOC + H] ⁺ ; m / z 328 (C ₁₆ H ₂₀ NO ₇ S; [M + H ] ⁺ ; M / z 428).

Reference Example 3: 4-hydroxymethyl-2,6-dimethoxyphenylboronic acid (S3)
An anhydrous tetrahydrofuran (1 L) solution of (3,5-dimethoxyphenyl) methanol (53 g, 0.31 mol, Aldrich) was cooled to -50 ° C to -70 ° C under argon, and n-butyllithium (450 mL, hexane 1.6M, 0.72 mol) for 30 minutes. After the addition, the reaction mixture was warmed to 0 ° C. over 45 minutes and then allowed to stand at room temperature for 2 hours. Next, the reaction system was cooled again to −60 ° C., and trimethyl borate (135 mL, 1.15 mol) was added dropwise little by little. After the addition, the mixture was warmed to room temperature and stirred for an additional 18 hours. The reaction was quenched at 0 ° C. by the addition of an aqueous citric acid solution (75 g of citric acid in 300 mL of water). Sodium chloride was added to saturate the aqueous layer and the product was extracted with ethyl acetate (2 × 1 L). The combined organic layers were dried over magnesium sulfate, filtered and concentrated under reduced pressure. Ethyl acetate (100 mL) was added to the residue, and the resulting colorless precipitate was collected by filtration and dried at 40 ° C. under reduced pressure to give the title compound as a colorless solid. This was used in the following reaction without purification.

Reference Example 4: (S) -2-tert-butoxycarbonylamino-3- (4′-hydroxymethyl-2 ′, 6′-dimethoxybiphenyl-4-yl) propionic acid methyl ester (S4)
Triethylamine (28 mL, 0.20 mol) was added to (S) -2-tert-butoxycarbonylamino-3- (4-trifluoromethanesulfonyloxyphenyl) propionic acid methyl ester (S2, 42.73 g, 0.10 mol) and 4-hydroxy Methyl-2,6-dimethoxyphenylboronic acid (S3, 29.7 g, 0.14 mol) was added to a dry dimethylformamide (250 mL) solution. After degassing with argon, tetrakis (triphenylphosphine) palladium (0) (5.8 g, 5 mmol) was added and the mixture was heated at 90 ° C. under argon for 1 h [Caution: Slightly exothermic. ]. After cooling to room temperature, the mixture was diluted with ethyl acetate (1 L) and water (700 mL) and separated. The organic layer was washed with water (2 × 300 mL), dried over magnesium sulfate, filtered and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (Biotage 75L, silica 800 g, elution solvent; ethyl acetate: hexane (50:50)) to give the title compound as a colorless solid; LC / MS (ES + ve): [ M-BOC + H] ⁺ ; m / z 346 (C ₂₄ H ₃₁ NO ₇ ; [M + H] ⁺ ; m / z 446).

Reference Example 5: (S) -2-tert-butoxycarbonylamino-3- (4′-formyl-2 ′, 6′-dimethoxybiphenyl-4-yl) propionic acid methyl ester (S5)
Manganese dioxide (230 g, 2.64 mol) was converted to (S) -2-tert-butoxycarbonylamino-3- (4′-hydroxymethyl-2 ′, 6′-dimethoxybiphenyl-4-yl) propionic acid methyl ester (S4 , 28.98 g, 65.1 mmol) in dichloromethane (1 L) in small portions. The resulting suspension was stirred at room temperature for 1 hour, filtered through celite, and the celite pad was washed with dichloromethane (1 L). The filtrate was concentrated under reduced pressure to give the product as a colorless foam. This was used without purification. LC / MS (ES + ve): [M-BOC + H] ⁺ ; m / z 344 (C ₂₄ H ₂₉ NO ₇ : [M + H] ⁺ : m / z 444).

Reference Example 6: (S) -2-tert-butoxycarbonylamino-3- [4 ′-(hydroxyiminomethyl) -2 ′, 6′-dimethoxybiphenyl-4-yl] propionic acid methyl ester (S6)
Hydroxylamine hydrochloride (7.4 g, 106 mmol) and diisopropylethylamine (18 mL, 106 mmol) were added to (S) -2-tert-butoxycarbonylamino-3- (4'-formyl-2 ', 6'-dimethoxybiphenyl) -4-yl) propionic acid methyl ester (S5, 23.6 g, 53 mmol) was added to a tetrahydrofuran (300 mL) solution. The reaction mixture was heated to reflux for 2 hours and then cooled to room temperature. The solution was concentrated under reduced pressure, then redissolved in ethyl acetate (500 mL), washed with 10% aqueous citric acid, water and brine (500 mL each), dried over magnesium sulfate, filtered, and concentrated under reduced pressure to give a colorless foam. A product was obtained. This was used in the next step without purification. LC / MS (ES + ve): [M-BOC + H] ⁺ ; m / z 359 (C ₂₄ H ₃₀ N ₂ O ₇ ; [M + H] ⁺ ; m / z 459).

Reference example 7: (S) -2-amino-3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) propionic acid methyl ester hydrochloride (S7)
Thionyl chloride (30 mL, 411 mmol) methyl (S) -2-tert-butoxycarbonylamino-3- [4 '-(hydroxyiminomethyl) -2', 6'-dimethoxybiphenyl-4-yl] propionate To a solution of ester (S6, 46.5 g, 101 mmol) in dichloromethane (500 mL) was added dropwise at 0 ° C. under argon. The reaction system was warmed to room temperature and stirred for 3 days. The precipitated solid was collected by filtration, washed with dichloromethane (200 mL), and dried at 45 ° C. under reduced pressure. The title compound was isolated as a white solid. LC / MS (ES + ve): [M + H] ⁺ ; m / z 341 (C ₁₉ H ₂₀ N ₂ O ₄ ; [M + H] ⁺ ; m / z 341).

Reference Example 8: 2-Bromo-5-hydroxybenzoic acid methyl ester (S8)
Add 2-bromo-5-methoxybenzoic acid in anhydrous dichloromethane (600 mL) cooled with dry ice / acetone while adding boron tribromide (1M in dichloromethane, 280 mL, 280 mmol) over 15 min under argon. 30.0 g, 130 mmol, Aldrich). The mixture was then stirred at room temperature for 2.5 hours to obtain a precipitate. Methanol (300 mL) was then carefully added dropwise over 30 minutes (Caution: first boiled with strong exotherm), finally resulting in a homogeneous dark solution. To this was added concentrated sulfuric acid (15 mL). The solution is stirred at reflux for 1 hour, cooled and concentrated in vacuo. The residue was dissolved in dichloromethane (500 mL), washed with brine (500 mL), dried over magnesium sulfate, filtered and concentrated in vacuo to give the title compound as a solid. LC / MS (ES + ve): [M + H] ⁺ ; m / z 231, 233 (C ₈ H ₇ BrO ₃ ; [M + H] ⁺ ; m / z 231, 233).

Reference Example 9: 2-Bromo-5-ethoxybenzoic acid methyl ester (S9)
Sodium hydride (mineral oil) in anhydrous dimethylformamide (150 mL) cooled in ice while adding 2-bromo-5-hydroxybenzoic acid methyl ester (S8, 20.41 g, 88 mmol) over 15 min under argon Medium 60%, 4.24 g, 106 mmol). The mixture was stirred at ambient temperature for 45 minutes, cooled again in ice and treated with iodoethane (8.5 mL, 106 mmol). The mixture was stirred at room temperature for 3 hours, concentrated in vacuo, diluted with ethyl acetate (400 mL), washed with water (3 × 400 mL) and brine (400 mL), dried over magnesium sulfate, filtered, Concentration in vacuo gave the title compound. This was contaminated with a small amount of the corresponding ethyl ester and residual mineral oil. This material was used in the next step without purification; LC / MS (ES + ve): [M + H] ⁺ ; m / z 259, 261 (C ₁₀ H ₁₁ BrO ₃ ; [M + H] ⁺ M / z 259, 261).

Reference Example 10: 2-Bromo-5-ethoxybenzoic acid (S10)
2-Bromo-5-ethoxybenzoic acid methyl ester (S9, 20.53 g, 79 mmol) is stirred in tetrahydrofuran (600 mL) and treated with lithium hydroxide (18.2 g, 791 mmol) in water (200 mL). did. The mixture was stirred for 4 days, concentrated in vacuo, diluted with water (500 mL), acidified with 5N hydrochloric acid, and extracted with ethyl acetate (2 × 300 mL). The combined extracts were washed with brine (250 mL), dried over magnesium sulfate, filtered and concentrated in vacuo to give an off-white solid. Triturated in hexane, filtered and dried to give the title compound as a white powder. LC / MS (ES + ve): [M + H] ⁺ ; m / z 245, 247 (C ₉ H ₉ BrO ₃ ; [M + H] ⁺ ; m / z 245, 247).

Reference Example 11: (S) -2-{[1- (2-Bromo-5-ethoxyphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) Propionic acid methyl ester (S11)
Under argon, (S) -2-amino-3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) propionic acid methyl ester hydrochloride (S7, 37.1 g, 98.5 mmol) was added to dichloromethane ( (500 mL) and water (400 mL) and cooled to 0 ° C. Sodium bicarbonate (20.1 g, 239.3 mmol) was added to the reaction mixture, followed by 2-bromo-5-ethoxybenzoyl chloride (27.2 g, 103.7 mmol) (oxalyl chloride in dichloromethane (4 eq) and 1 drop of dimethylformamide) 2-Bromo-5-ethoxybenzoic acid (S10) prepared by standard means) was added dropwise. The reaction was stirred at 0 ° C. for 1 hour and then diluted with a saturated aqueous solution of sodium bicarbonate (200 mL). After separating the organic layer, the aqueous layer was extracted again with dichloromethane (2 × 400 mL). The combined organic layers were dried over sodium sulfate, filtered and concentrated under reduced pressure. The product was purified by silica gel chromatography (Biotage 75L, silica 800 g, elution solvent; ethyl acetate: dichloromethane (3:97)) to give the title compound as a colorless solid: MS (ES + ve): [M + H ^{] +; m / z 567,} 569 (C 28 H 27 BrN 2 O 6; [M + H] +; m / z 567, 569).

Reference Example 12: (S) -2-{[1- (2-Bromo-5-methoxyphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) Propionic acid ethyl ester (S12)
2-Bromo-5-methoxybenzoic acid (0.355 g, 1.54 mmol, Aldrich), O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium under argon Hexafluorophosphate (1.168 g, 2 eq) and triethylamine (1.069 mL, 5 eq) were added to dimethylformamide (25 mL) with stirring at room temperature. After 0.5 hour, ethyl ester corresponding to S7 (S13), (S) -2-amino-3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) propionic acid ethyl ester hydrochloride ( 0.6 g, 1 eq) was added and stirring was continued overnight. The solvent was then concentrated under reduced pressure and the residue was partitioned between ethyl acetate and water. The organic layer was washed with water (x2) and saturated aqueous sodium bicarbonate and then concentrated to dryness. The crude product was purified by silica-column chromatography (elution solvent: 0-10% methanol gradient in dichloromethane) and then the title compound was obtained by preparative HPLC.
MS (AP + ve): [M + H] ⁺ ; m / z 567, 569 (C ₂₈ H ₂₇ BrN ₂ O ₆ ; [M + H] ⁺ ; m / z 567, 569).

Reference Example 13: (S) -2-Amino-3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) propionic acid ethyl ester hydrochloride (ethyl ester corresponding to S7) (S13)
Under argon, (S) -2-tert-butoxycarbonylamino-3- [4 '-(hydroxyiminomethyl) -2', 6'-dimethoxybiphenyl-4-yl] propionic acid ethyl ester (12.5 g, 26.5 mmol ) [Prepared in the same manner as S1-S6 except that Reference Example 1 was started with L-tyrosine ethyl ester hydrochloride (from Bachem) instead of L-tyrosine methyl ester] in dichloromethane (250 mL) To the solution was added thionyl chloride (7.71 mL, 105.7 mmol) dropwise at 0 ° C. over 15 minutes. The reaction was stirred at batch temperature for an additional 15 minutes, then warmed to room temperature and continued to stir overnight. The precipitated title compound was collected by filtration, washed with dichloromethane (2 × 15 mL), and dried under reduced pressure. LC / MS (ES + ve): [M + H] ⁺ ; m / z 355 (C ₂₀ H ₂₂ N ₂ O ₄ ; [M + H] ⁺ ; m / z 355).

(S)-2-{[1-(2-ブロモ-5-エトキシフェニル)メタノイル]アミノ}-3-(4'-シアノ-2',6'-ジメトキシビフェニル-4-イル)プロピオン酸メチルエステル(Ｓ１１, 51.5 g, 90.8 mmol)のテトラヒドロフラン(950 mL)溶液を０℃まで冷却し、0.5M 水酸化リチウム水溶液(700 mL)で処理した。反応混合物を０℃で１時間攪拌した後、5M塩酸で酸性にした。テトラヒドロフランを減圧下留去し残渣を酢酸エチルで希釈した。有機層を分離した後、水相を酢酸エチルで再抽出した。合した有機層を水で洗浄し(×２)、硫酸マグネシウムで乾燥し、溶媒を減圧下留去して表題化合物を無色固体として得た；
¹H NMR δ (DMSO-d6): 1.29 (3H, t, J 7.0Hz), 2.96 (1H, dd, J 13.9, 10.9Hz), 3.22 (1H, dd, J 14.1, 4.2Hz), 3.71 (6H, s), 3.99 (2H, q, J 7.0Hz), 4.65 (1H, ddd, J 10.9, 8.4, 4.3Hz), 6.69 (1H, d, J 3.0Hz), 6.91 (1H, dd, J 8.8, 3.1Hz), 7.14 (2H, d, J 8.1Hz), 7.24 (2H, s), 7.32 (2H, d, J 8.1Hz,), 7.47 (1H, d, J 8.8Hz), 8.77 (1H, br. d, J 8.4Hz), 12.83 (1H, br. s); MS (ES+ve) [M+H]⁺；m/z 553, 555 (C₂₇H₂₅BrN₂O₆；[M+H]⁺；m/z 553, 555)。 Production Example 1: (S) -2-{[1- (2-bromo-5-ethoxyphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) Propionic acid (P1)

(S) -2-{[1- (2-Bromo-5-ethoxyphenyl) methanoyl] amino} -3- (4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl) propionic acid methyl ester A solution of (S11, 51.5 g, 90.8 mmol) in tetrahydrofuran (950 mL) was cooled to 0 ° C. and treated with 0.5 M aqueous lithium hydroxide (700 mL). The reaction mixture was stirred at 0 ° C. for 1 hour and then acidified with 5M hydrochloric acid. Tetrahydrofuran was distilled off under reduced pressure, and the residue was diluted with ethyl acetate. After separating the organic layer, the aqueous phase was re-extracted with ethyl acetate. The combined organic layers were washed with water (x2), dried over magnesium sulfate, and the solvent was removed under reduced pressure to give the title compound as a colorless solid;
¹ H NMR δ (DMSO-d6): 1.29 (3H, t, J 7.0Hz), 2.96 (1H, dd, J 13.9, 10.9Hz), 3.22 (1H, dd, J 14.1, 4.2Hz), 3.71 (6H , s), 3.99 (2H, q, J 7.0Hz), 4.65 (1H, ddd, J 10.9, 8.4, 4.3Hz), 6.69 (1H, d, J 3.0Hz), 6.91 (1H, dd, J 8.8, 3.1Hz), 7.14 (2H, d, J 8.1Hz), 7.24 (2H, s), 7.32 (2H, d, J 8.1Hz,), 7.47 (1H, d, J 8.8Hz), 8.77 (1H, br d, J 8.4Hz), 12.83 (1H, br. s); MS (ES + ve) [M + H] ⁺ ; m / z 553, 555 (C ₂₇ H ₂₅ BrN ₂ O ₆ ; [M + H ] ⁺ ; M / z 553, 555).

Production Example 2: (S) -2-{[1- (2-Bromo-5-fluorophenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) Propionic acid (P2)
The title compound was produced in the same manner as described in Reference Example 11 and Production Example 1 using S13 and 2-bromo-5-fluorobenzoyl chloride (Apollo). [M + H] ⁺ ; m / z 527,529.

Production Example 3: (S) -2-{[1- (2-Bromo-5-methoxyphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) Propionic acid (P3)
Aqueous lithium hydroxide (0.5 M, 25 mL) was added to (S) -2-{[1- (2-bromo-5-methoxyphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6 '-Dimethoxybiphenyl-4-yl) propionic acid ethyl ester (S12, 0.539 g, 0.95 mmol) was added to a tetrahydrofuran (30 mL) solution with stirring at room temperature. Stirring was continued for 2 hours and then the reaction was quenched with excess 10% aqueous citric acid. The mixture was then extracted with ethyl acetate and the organic layer was washed with fresh citric acid followed by water (twice). The organic layer was concentrated to dryness and the resulting crude product was purified by MDAP to give the title compound as a white solid. [M + H] ⁺ ; m / z 539, 541.

Production Example 4: (S) -2-{[1- (2-bromo-5-methylphenyl) methanoyl] amino} -3- (4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl) Propionic acid (P4)
Using S13 and 2-bromo-5-methylbenzoic acid (Apin), the ethyl ester obtained by the same method as described in Reference Example 12 was hydrolyzed by the method described in Preparation Example 3 to give the title compound. Manufactured. [M + H] ⁺ ; m / z 523, 525.

Production Example 5: (S) -2-{[1- (2-Bromo-5-chlorophenyl) methanoyl] amino} -3- [4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl] propion Acid (P5)
The title compound was produced from the corresponding ethyl ester in the same manner as described in Production Example 3. The ethyl ester was prepared from S13 and 2-bromo-5-chlorobenzoic acid (Lancaster) by the method described in Reference Example 12. [MH] ^- ; m / z 541, 543, 545.

Production Example 6: (S) -2-{[1- (2,5-dibromophenyl) methanoyl] amino} -3- [4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl] propionic acid (P6)
The title compound was produced from the corresponding ethyl ester in the same manner as described in Production Example 3. The ethyl ester was prepared from S13 and 2,5-dibromobenzoic acid (Lancaster) by the method described in Reference Example 12. [MH] ^- ; m / z 585, 587, 589.

Production Example 7: (S) -2-{[1- (5- (iso-propoxy) -2-bromophenyl) methanoyl] amino} -3- [4′-cyano-2 ′, 6′-dimethoxybiphenyl- 4-yl] propionic acid (P7)
The title compound was produced from the corresponding ethyl ester in the same manner as described in Production Example 3. The ethyl ester is a compound of S13 and 5-iso-propoxy-2-bromobenzoic acid (except for using iso-propyl bromide in an alkylation step similar to that of Reference Example 9 and the procedures of Reference Example 8 to Reference Example 10). In the same manner, it was produced by the method described in Reference Example 12. [MH] ^- , m / z 565, 567.

Formula compounds and pharmaceutical compositions of the present invention containing a pharmacologically acceptable derivative thereof as an active ingredient of formula (I) are, alpha ₄ integrin therapy mediated cell adhesion various diseases inhibition is effective or It can be used for prevention, for example, rheumatoid arthritis, asthma, allergic diseases, adult respiratory distress syndrome, AIDS dementia, Alzheimer's disease, cardiovascular disease, thrombosis or harmful platelet aggregation, reocclusion after thrombolysis, resuscitation Perfusion injury, skin inflammatory disease, diabetes, multiple sclerosis, systemic lupus lupus erythematosus, inflammatory bowel disease, disease involving leukocyte infiltration into the gastrointestinal tract, disease involving leukocyte infiltration into epithelial tissue, pancreatitis, mammary gland Inflammation, hepatitis, cholecystitis, cholangitis, cholangitis, bronchitis, sinusitis, pulmonary inflammatory disease, collagen disease, sarcoidosis, osteoporosis, osteoarthritis, atherosclerosis Pulse sclerosis, neoplastic disease, trauma, eye disease, Sjogren's syndrome, rejection after organ transplantation, host versus graft or graft versus host disease, intimal thickening, arteriosclerosis, post-operative reinfarction or restenosis, It can be applied to the treatment or prevention of nephritis, tumor angiogenesis, malignant tumor, multiple myeloma and myeloma-induced bone resorption, sepsis, central neuropathy, Meniere disease and inflammatory bowel disease.

Claims (10)

Formula (I):

(I)
(Wherein R ¹ represents bromo, R ² represents halogen, C _1-6 alkyl or C _1-6 alkoxy)
Or a pharmacologically acceptable derivative thereof as an active ingredient. The pharmaceutical composition according to claim 1, wherein R ² is halogen or C _1-6 alkoxy. The pharmaceutical composition according to claim 2, wherein R ² is fluoro, methoxy or ethoxy. As an active ingredient, (S) -2-{[1- (2-bromo-5-methylphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) Propionic acid,
(S) -2-{[(2-bromo-5-chlorophenyl) methanoyl] amino} -3- [4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl] propionic acid,
(S) -2-{[(2,5-dibromophenyl) methanoyl] amino} -3- [4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl] propionic acid,
(S) -2-{[(5- (iso-propoxy) -2-bromophenyl) methanoyl] amino} -3- [4'-cyano-2 ', 6'-dimethoxybiphenyl-4-yl] propionic acid The pharmaceutical composition according to claim 1, comprising a compound selected from the group consisting of: or a pharmacologically acceptable derivative thereof.   As an active ingredient, (S) -2-{[1- (2-bromo-5-ethoxyphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) A pharmaceutical composition comprising propionic acid or a pharmacologically acceptable derivative thereof.   As an active ingredient, (S) -2-{[1- (2-bromo-5-fluorophenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) A pharmaceutical composition comprising propionic acid or a pharmacologically acceptable derivative thereof.   As an active ingredient, (S) -2-{[1- (2-bromo-5-methoxyphenyl) methanoyl] amino} -3- (4′-cyano-2 ′, 6′-dimethoxybiphenyl-4-yl) A pharmaceutical composition comprising propionic acid or a pharmacologically acceptable derivative thereof. The pharmaceutical composition according to claim 1 inhibition of alpha ₄ integrin mediated cell adhesion is a therapeutic or prophylactic agent for beneficial conditions.   Rheumatoid arthritis; asthma; rhinitis; adult respiratory distress syndrome; AIDS dementia; Alzheimer's disease; thrombosis; reocclusion after thrombolysis; reperfusion injury; psoriasis; eczema; contact dermatitis; atopic dermatitis; Multiple sclerosis; systemic lupus lupus erythematosus; ulcerative colitis; Crohn's disease; cystitis; pediatric steatosis; nontropical sprue; lymphocytic colitis; collagenous colitis; eosinophilic gastroenteritis; Mastitis; hepatitis; cholecystitis; cholangitis; perichonditis; bronchitis; sinusitis; hypersensitivity pneumonia; collagen disease; sarcoidosis; osteoporosis; osteoarthritis; atherosclerosis; Exfoliation; allergic conjunctivitis; autoimmune uveitis; Sjogren's syndrome; rejection after organ transplant; host versus graft or graft versus host disease; intimal thickening; arteriosclerosis; percutaneous transluminal coronary artery Reinfarction or restenosis after surgery such as angioplasty (PTCA) or percutaneous transluminal recanalization; nephritis; tumor angiogenesis; malignant tumor; multiple myeloma; myeloma-induced bone resorption; sepsis; The pharmaceutical composition according to any one of claims 1 to 8, which is a pathological condition selected from traumatic brain injury; traumatic spinal cord injury; Meniere's disease and inflammatory bowel disease.   The pharmaceutical composition according to claim 9, wherein the pathological condition is asthma, rheumatoid arthritis, inflammatory bowel disease or multiple sclerosis.