JP2005068145A - Pharmaceutical composition - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a pharmaceutical composition useful for treating or preventing an α<SB>L</SB>β<SB>2</SB>integrin-mediated pathologic state and a substance P-related pathologic state. <P>SOLUTION: This pharmaceutical composition contains a compound expressed by formula (I) (R is H, hydroxyl or carbamoyl; and n is 1 or 2) or a pharmacologically acceptable salt thereof as an active ingredient. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

The present invention relates to a pharmaceutical composition comprising a novel compound having an excellent α _L β ₂ integrin-mediated cell adhesion inhibitory action and substance P receptor antagonistic action as an active ingredient.

Leukocyte integrins and intercellular adhesion molecules (ICAM) play a central role in leukocyte adhesion to target cells and extracellular matrix. The β ₂ integrin (CD18) subfamily has four members: α _L β ₂ integrin (LFA-1, CD11a / CD18), α _M β ₂ integrin (Mac-1, CD11b / CD18), α _X It consists of β ₂ integrin (p150 / 95, CD11c / CD18) and α _D β ₂ integrin (CD11d / CD18) (Non-Patent Document 1), which are non-associated with CD18 and a related α chain. Consists of covalent bonds. In particular, LFA-1 has been shown to be a major factor in cell adhesion of T cells, eosinophils and other leukocytes and invasion of extravascular inflammatory tissues (Non-patent Document 2; Non-patent Document 3). . LFA-1 binds to the ICAM family (ICAM-1, 2, 3, 4, 5) found in many cell types such as vascular endothelial cells, dendritic neurons, epithelial cells, macrophages and T lymphoblasts. (Non-Patent Document 4). Furthermore, the interaction of LFA-1 / ICAM-1 and LFA-1 / ICAM-3 can function as a co-stimulatory signal required for T cell activation (Non-patent Document 5).

  Cell invasion and T cell activation are important processes in many inflammatory disease states. In animal models of several different inflammatory diseases, antibodies against LFA-1 and ICAM-1 suppress inflammation, indicating that LFA-1 has an important role in inflammation (non-patented) Reference 6; Non-patent document 7; Non-patent document 8). Furthermore, humanized monoclonal antibodies against CD11a (alpha chain of LFA-1) have shown efficacy in patients with psoriasis (Non-Patent Document 9).

Furthermore, it is known that antibodies against LFA-1 suppress rejection after organ transplantation (Non-patent Documents 10 and 11). In addition, use of a monoclonal antibody against α _L β ₂ integrin for transplantation is also disclosed (Patent Document 1).

  On the other hand, substances having an antagonistic action on the substance P receptor (ie, neurokinin 1 receptor) are inflammatory diseases (for example, asthma, rheumatoid arthritis, inflammatory bowel disease, cystitis and other gastric disorders). It is thought that it is useful for the treatment of non-patent document 12, non-patent document 13, and non-patent document 14.

International Publication No. 94/04188 Specification Bokner et al., Adhesion Molecules in Allergic Disease, Marcel Dekker, Inc. pp 1-24 (1997) Garmberg, Curr. Opin. Cell Biology, 9, 643-650 (1997) Pains et al., Br. J. Pharmacology, 126, 537-550 (1999) Dustin et al., J. Immunology, 137, 245-254 (1986) Wingren et al., Crit. Rev. in Immunology, 15, 235-253 (1995) Rothlein et al., Kidney International, 41, 617 (1992) Igo et al., J. Immunology, 147, 4167 (1991) Bennet et al., J. Pharmacol. And Exp. Therapeutics, 280, 988 (1997) Gottlieb et al., J. Am. Acad. Dermatology, 42, 428-35 (2000) Poston et al., Transplantation 69, 2005-2013 (2000) Nakakura et al., Transplantation 62, 547-552 (1996) Kleinbeld et al., Int. Immunopharmacology, 1, 1629-1650 (2001) Swain et al., Ann. Rep. Med. Chem., 34, 51-60 (1999) Onmahatto Jr. et al., Ann. Rep. Med. Chem., 33, 71-80 (1998)

（式中、Ｒは水素原子、ヒドロキシル基またはカルバモイル基、ｎは１または２を表す）
で示される化合物、またはその薬理学的に許容される塩を有効成分とする医薬組成物に関する。 The present invention relates to a compound of formula (I):

(Wherein R represents a hydrogen atom, a hydroxyl group or a carbamoyl group, and n represents 1 or 2)
Or a pharmacologically acceptable salt thereof as an active ingredient.

  The active ingredient of the present invention includes optical isomers based on the asymmetric carbon, and the present invention also includes these isomers and mixtures thereof.

  In embodiments of the present invention, the configuration of the active ingredient bonds need not be fixed. The active ingredient of the present invention may be a compound having a single configuration or a mixture of several having different configurations.

In a preferred embodiment of active ingredient (I), R is a hydrogen atom.
In another preferred embodiment of active ingredient (I), R is a hydroxyl group.
In another preferred embodiment of active ingredient (I), R is a carbamoyl group.
In another preferred embodiment of active ingredient (I), n is 1.
In another preferred embodiment of active ingredient (I), n is 2.
In a more preferred embodiment of active ingredient (I), R is a hydrogen atom and n is 1.
In a more preferred embodiment of active ingredient (I), R is a hydroxyl group and n is 1.
In a more preferred embodiment of active ingredient (I), R is a carbamoyl group and n is 2.

A particularly preferable compound as an active ingredient of the pharmaceutical composition of the present invention is a compound selected from the following group.
(5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-acetylamino-1,3-diazabicyclo [3.3.0] octane-2,4-dione,
(5S, 7S) -5- (4-Cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(2-hydroxyacetyl) amino] -1,3-diazabicyclo [3.3.0] octane -2,4-dione and (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(3-carbamoylpropionyl) amino] -1,3-diazabicyclo [3.3.0] Octane-2,4-dione.

  The active ingredient of the present invention is characterized in that an acylamino group is bonded to the 7-position of the 1,3-diazabicyclo [3.3.0] octane skeleton, and a 4-cyanobenzyl group is bonded to the 5-position. There are no literatures describing compounds with characteristics.

  The active ingredient of the present invention has a strong inhibitory activity against both LFA-1-mediated cell adhesion and LFA-1-mediated T cell simultaneous activation. The active ingredient of the present invention also has an overall improvement in (a) plasma protein binding, (b) water solubility, and (c) logP (octanol / water partition rate), which makes it an excellent biomarker for oral administration. Indicates availability. Therefore, the active ingredient of the present invention exhibits an excellent effect in vivo against diseases involving LFA-1-mediated cell adhesion.

  Furthermore, the active ingredient of the present invention also has a strong antagonistic activity against the substance P receptor (ie, neurokinin 1 receptor). Substance P receptor antagonists are useful for the treatment of inflammatory diseases such as asthma, rheumatoid arthritis, inflammatory bowel disease, cystitis and other gastric disorders (Non-Patent Documents 12-14). Therefore, the active ingredient of the present invention has an excellent therapeutic effect on diseases caused by substance P. In addition, the active ingredient of the present invention can exhibit an excellent effect on the treatment or prevention of inflammatory diseases based on two activities of LFA-1-mediated cell adhesion inhibition and substance P receptor antagonism.

  Furthermore, since the active ingredient (I) of the present invention has lower cytotoxicity and lower cytochrome P450 inhibitory activity than the compounds described in the prior art, the risk of side effects is low.

  The active ingredient of the present invention can be used for the prevention / treatment of diseases in either a free form or a pharmacologically acceptable salt form. Examples of pharmacologically acceptable salts include acid addition salts with inorganic or organic acids (for example, hydrochloride, sulfate, nitrate, hydrobromide, methanesulfonate, p-toluenesulfonate, Acetate), and salts with inorganic bases, organic bases, or amino acids (for example, triethylamine salts, salts with lysine, salts with alkali metals, salts with alkaline earth metals, etc.). Further, pharmacologically acceptable salts include these intramolecular salts, solvates or hydrates.

  The pharmaceutical composition of the present invention is formulated from a therapeutically effective amount of the active ingredient (I) and a pharmaceutically acceptable carrier or diluent. Examples of pharmacologically acceptable carriers or diluents include binders (syrup, gum arabic, gelatin, sorbit, tragacanth, polyvinylpyrrolidone), excipients (lactose, sucrose, corn starch, potassium phosphate, sorbit) Glycine), lubricants (magnesium stearate, talc, polyethylene glycol, silica), disintegrants (potato starch) and wetting agents (sodium lauryl sulfate).

  The active ingredient of the present invention can be administered orally or parenterally, and therefore can be administered as a suitable pharmaceutical preparation. Suitable pharmaceutical preparations for oral administration include, for example, solid preparations such as tablets, granules, capsules and powders, or liquid preparations such as solution preparations, suspension preparations and emulsion preparations. Appropriate pharmaceutical preparations for parenteral administration include suppository forms, or injection preparations or intravenous infusion preparations using distilled water for injection, physiological saline, aqueous dextrose, etc., and conventional inhalants Is mentioned.

  The dose of the active ingredient of the present invention varies depending on the administration method, the patient's age, sex, body weight and medical condition, but in general, the dose per day is preferably about 0.1 to 100 mg / kg, particularly preferably. Is in the range of 1-100 mg / kg.

  The pharmaceutical composition of the present invention can be used as a therapeutic or prophylactic agent for a disease state associated with LFA-1-mediated cell adhesion. In addition, the compound of the present invention can be used as a therapeutic agent for patients suffering from or susceptible to pathologies associated with LFA-1-mediated cell adhesion. Examples of pathologies associated with LFA-1 mediated cell adhesion include inflammatory diseases, autoimmune diseases and allergic diseases.

  The pharmaceutical composition of the present invention is also used as a therapeutic or preventive agent for a disease state caused by or mediated by substance P in a patient, and as a therapeutic agent for a patient suffering from or susceptible to such a disease state. Can do. Examples of such pathological conditions include inflammatory diseases.

  The pharmaceutical composition of the present invention comprises rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, allergic disease, adult respiratory distress syndrome, AIDS, cardiovascular disease, thrombosis, harmful platelet aggregation, reocclusion after thrombolysis, reperfusion injury , Skin inflammatory diseases (eg psoriasis, eczema, contact dermatitis, atopic dermatitis), osteoporosis, osteoarthritis, atherosclerosis, arteriosclerosis (including transplanted arteriosclerosis after transplantation), new Biological diseases (including neoplastic or cancerous growth metastases), trauma, retinal detachment, type I diabetes, multiple sclerosis, systemic lupus erythematosus (SLE), ocular inflammatory conditions, inflammatory bowel disease (Crohn's disease, ulcerative) Such as colitis), cystitis, gastric disorder, localized ileitis, Sjogren's syndrome, and other autoimmune diseases.

  The pharmaceutical compositions of the invention also include transplant rejection, such as organ transplantation (ie, chronic rejection and acute rejection), and allograft rejection (host versus graft disease) and graft versus host disease. Can also be used.

  The pharmaceutical composition of the present invention preferably contains psoriasis, rheumatoid arthritis (or rheumatoid arthritis), inflammatory bowel disease (Crohn's disease, ulcerative colitis), systemic lupus erythematosus, atopic dermatitis, Sjogren's syndrome, post transplantation It can be used as a therapeutic and prophylactic agent for rejection (allograft rejection and graft-versus-host disease).

  The pharmaceutical composition of the present invention can also be suitably used as a therapeutic or prophylactic agent for inflammatory diseases such as asthma, inflammatory bowel disease, cystitis and other gastric disorders.

（式中、ｎは前記と同一意味を有する）
で示される化合物またはその薬理学的に許容される塩は、式（ＩＩ）：

で示される化合物を、式(ＩＩＩ−ａ):
Ｈ−（ＣＨ₂)_n−ＣＯＯＨ （ＩＩＩ−ａ）
（式中、ｎは前記と同一意味を有する）
で示される化合物またはその反応性誘導体と縮合させ、次いで、必要に応じて得られた化合物を薬理学的に許容される塩に変換することにより製造することができる。 The active ingredient of the present invention can be produced, for example, by the following method.
Manufacturing method A
Of the compounds represented by formula (I) which are active ingredients, formula (Ia):

(Wherein n has the same meaning as above)
Or a pharmaceutically acceptable salt thereof is represented by the formula (II):

A compound represented by formula (III-a):
_{H- (CH 2) n -COOH (} III-a)
(Wherein n has the same meaning as above)
Or a reactive derivative thereof, and then, if necessary, the compound obtained can be converted into a pharmacologically acceptable salt.

  The condensation reaction between compound (II) and compound (III-a) is carried out in the presence or absence of a base (eg, DIEA, triethylamine, pyridine, DMAP, etc.) in a suitable solvent (eg, ethyl acetate, dichloromethane, tetrahydrofuran). , Dimethylformamide, etc.) and a conventional condensing agent (eg, BOP-Cl, BOP reagent or EDC) can be used in the presence of an activator (eg, HOBT).

  In addition, the condensation reaction between the compound (II) and a reactive derivative of the compound (III-a) (for example, an acid halide such as acid chloride or an acid anhydride) is performed by using a base (for example, DIEA, triethylamine, pyridine, DMAP, etc.) Can be carried out at a temperature of −30 ° C. to room temperature in a suitable solvent (for example, ethyl acetate, dichloromethane, tetrahydrofuran, dimethylformamide, etc.) or without solvent.

（式中、ｎは前記と同一意味を有する）
で示される化合物またはその薬理学的に許容される塩は、式（ＩＩ）で示される化合物を、式（ＩＩＩ−ｂ）：
Ｒ¹Ｏ−（ＣＨ₂）_n−ＣＯＯＨ (ＩＩＩ−ｂ)
（式中、Ｒ¹Ｏは保護されたヒドロキシル基を表し、ｎは前記と同一意味を有する）
で示される化合物またはその反応性誘導体と縮合させ、保護基を除去し、次いで必要に応じて、得られた化合物を薬理学的に許容される塩に変換することにより製造することができる。 Manufacturing method B
Of the compounds represented by formula (I) which are active ingredients, formula (Ib):

(Wherein n has the same meaning as above)
The compound represented by the formula (II) or a pharmacologically acceptable salt thereof is obtained by converting the compound represented by the formula (II) into the formula (III-b):
^{_{R 1 O- (CH 2) n}} -COOH (III-b)
(Wherein R ¹ O represents a protected hydroxyl group, and n has the same meaning as described above)
Or a reactive derivative thereof, to remove the protecting group, and then, if necessary, convert the obtained compound into a pharmacologically acceptable salt.

  The protecting group for the hydroxyl group can be selected from usual protecting groups for the hydroxyl group (for example, trimethylsilyl group, benzyl group, methyl group, etc.) that can be easily removed according to a conventional method.

  The condensation reaction of compound (II) with compound (III-b) or a reactive derivative thereof can be carried out in the same manner as described in the above-mentioned production method A.

Deprotection can be carried out according to a conventional method such as hydrolysis, acid treatment, BBr ₃ treatment, and catalytic reduction according to the type of protecting group to be removed.

  The catalytic reduction can be carried out under a hydrogen atmosphere using a catalyst such as palladium-carbon in a suitable solvent of acetic acid (for example, methanol, ethanol, acetic acid) at room temperature to under heating.

Demethylation using BBr ₃ can be performed in a suitable solvent (eg, tetrahydrofuran, dichloromethane, acetic acid) at a temperature of −78 ° C. to 50 ° C.

（式中、ｎは前記と同一意味を有する）
またはその薬理学的に許容される塩は、化合物(ＩＩ)を、式(ＩＩＩ−ｃ)：
Ｈ₂ＮＣ(＝Ｏ)−(ＣＨ₂)_n−ＣＯＯＨ (ＩＩＩ−ｃ)
（式中、ｎは前記と同一意味を有する）
で示される化合物またはその反応性誘導体と縮合させ、次いで、必要に応じて得られた化合物を薬理学的に許容される塩に変換することにより製造することができる。 Manufacturing method C
Of the target compound (I), the formula (Ic):

(Wherein n has the same meaning as above)
Or a pharmacologically acceptable salt thereof, compound (II), compound of formula (III-c):
_{H 2 NC (= O) -} (CH 2) n -COOH (III-c)
(Wherein n has the same meaning as above)
Or a reactive derivative thereof, and then, if necessary, the compound obtained can be converted into a pharmacologically acceptable salt.

  The condensation reaction of compound (II) with compound (III-c) or a reactive derivative thereof can be carried out in the same manner as described in Production Method A.

  The compound of the formula (II) can be produced according to the method described in WO01 / 30781 or the method described in Reference Examples described later.

In the present specification and claims, C _1-6 alkyl means a linear or branched alkyl group having 1 to 6 carbon atoms, such as a methyl group, an ethyl group, a propyl group, an isopropyl group, A butyl group, an isobutyl group, etc. are mentioned, Preferably it has 1-4 carbon atoms.

Abbreviations:
BOP-Cl: bis (2-oxo-3-oxazolidinyl) phosphinic chloride BOP reagent: benzotriazol-1-yloxy-tris (dimethylamino) phosphonium hexafluorophosphate BSA: bovine serum albumin DIEA: diisopropylethylamine DMAP: 4-dimethyl Aminopyridine EDC: 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride HBSS: Hank's balanced salt solution HOBT: 1-hydroxybenzotriazole HSA: human serum albumin PBS: phosphate buffered saline

Production Example 1: (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-acetylamino-1,3-diazabicyclo [3.3.0] octane-2, 4-dione (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-amino-1,3-diazabicyclo [3.3.0] octane-2,4- Acetic anhydride (1.0 mL) was added to a solution of dione (78.5 mg) in tetrahydrofuran (5 mL). The reaction mixture was stirred at 45 ° C. for 2 hours, the mixture was concentrated and purified by preparative thin layer chromatography (silica gel; dichloromethane) to give the title compound (84 mg). MS (m / z) 478.8 (MNa ⁺ ).

Production Example 2: (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(3-carbamoylpropionyl) amino] -1,3-diazabicyclo [3.3 .0] octane-2,4-dione (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-amino-1,3-diazabicyclo [3.3.0] ] A mixture of octane-2,4-dione (82.7 mg), succinamic acid (45.86 mg), EDC (93.12 mg), HOBT (61.24 mg), DIEA (104.79 μL) and tetrahydrofuran (5 mL). Stir overnight at room temperature. The reaction mixture was concentrated and purified by high performance liquid chromatography (HPLC) (Beckman 5μ C18 column; gradient elution: water / acetonitrile = 10-100% / 0.1% trifluoroacetic acid) to give 72 mg of the title compound. MS (m / z) 536 (MNa ⁺ ).

Production Example 3: (5S, 7S) -5- (4-Cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(2-carbamoylacetyl) amino] -1,3-diazabicyclo [3.3 .0] octane-2,4-dione (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-amino-1,3-diazabicyclo [3.3.0] A mixture of octane-2,4-dione (200 mg), malonamic acid (59.5 mg), EDC (112 mg), HOBT (97.5 mg), DIEA (168 μL) and tetrahydrofuran (5 mL) was stirred at room temperature overnight. . The reaction mixture was concentrated. The residue was dissolved in ethyl acetate and the resulting solution was washed, dried and concentrated to give the title compound (212 mg). MS (m / z) 500 (MH ^<+> ).

Production Example 4: (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(3-hydroxypropionyl) amino] -1,3-diazabicyclo [3.3 .0] octane-2,4-dione (1) (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-amino-1,3-diazabicyclo [3. 3.0] Octane-2,4-dione (0.300 g), 3-methoxypropionic acid (0.209 μL), EDC (0.224 g), HOBT (0.221 g), DIEA (0.38 μL) and tetrahydrofuran (15 mL) of the mixture was stirred overnight at room temperature. The reaction mixture was concentrated. The residue was purified by HPLC [Beckman 5 μC18 column; gradient elution: water / acetonitrile (10-100%) / 0.1% acetic acid] to give a foam. This was dissolved in ethyl acetate and the resulting solution was washed, dried and concentrated to (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7- [ (3-Methoxypropionyl) amino] -1,3-diazabicyclo [3.3.0] octane-2,4-dione was obtained (0.259 g). MS (m / z) 501 (MH ^<+> ).

(2) Boron tribromide (1M in dichloromethane, 3 mL) was converted to (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(3-methoxypropionyl) Amino] -1,3-diazabicyclo [3.3.0] octane-2,4-dione (0.16 g) was added to a solution of dichloromethane (15 mL) at −78 ° C., and the mixture was stirred for 8 hours at −78 ° C. did. The mixture was concentrated and the residue was purified by HPLC [Beckman 5 μC18 column; gradient elution: water / acetonitrile (10-100%) / 0.1% acetic acid)] to give a foam. This was dissolved in ethyl acetate and the resulting solution was washed, dried and concentrated to give the title compound (0.119 g). MS (m / z) 487 (MH ^<+> ).

Production Example 5: (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(2-hydroxyacetyl) amino] -1,3-diazabicyclo [3.3 .0] octane-2,4-dione (1) (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-amino-1,3-diazabicyclo [3. 3.0] To a solution of octane-2,4-dione (150 mg) and DIEA (189 μL) in tetrahydrofuran (4 mL) was added a solution of benzyloxyacetyl chloride (57 μL) in tetrahydrofuran (2 mL) and the mixture was stirred at room temperature overnight. did. The reaction mixture was concentrated and the residue was taken up in ethyl acetate. The resulting solution was washed, dried, filtered and concentrated. The residue was purified by HPLC [Beckman 5 μC18 column; gradient elution: water / acetonitrile (10-100%) / 0.1% acetic acid] to give a foam. This was dissolved in ethyl acetate and the resulting solution was washed, dried and concentrated to (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7- [ (2-Benzyloxyacetyl) amino] -1,3-diazabicyclo [3.3.0] octane-2,4-dione was obtained (0.135 g). MS (m / z) 563.4 [MH ^<+ >].

(2) (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(2-benzyloxyacetyl) amino] -1,3-diazabicyclo obtained above Hydrogen was bubbled into a mixture of [3.3.0] octane-2,4-dione (0.125 g), 5% palladium-carbon (15 mg) and ethanol (10 mL) and the resulting mixture was charged with hydrogen overnight. Stir under atmosphere. 5% Palladium-carbon (10 mg) was added to the mixture, and the mixture was stirred overnight under a hydrogen atmosphere. The reaction mixture was filtered through celite, and the filtrate was concentrated. The residue was purified by HPLC (Beckman 5 μC18 column; gradient elution: water / acetonitrile (10-100%) / 0.1% trifluoroacetic acid) to give the title compound (0.023 g). MS (m / z) 473 [MH ^<+ >] and 495 [MNa ^<+ >].

（上記の反応工程式において、ｔＢＤＭＳＯはｔｅｒｔ−ブチルジメチルシリルオキシ基であり、ＭｓＯはメタンスルホニルオキシ基である。） Reference example: (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-amino-1,3-diazabicyclo [3.3.0] octane-2,4- Dione The title compound was prepared according to the following reaction scheme.
Reaction process formula

(In the above reaction process formula, tBDMSO is a tert-butyldimethylsilyloxy group, and MsO is a methanesulfonyloxy group.)

(1) p-Toluenesulfonic acid (50.6 g) is added to a suspension of L-4-trans-hydroxyproline (25.25 g) in a mixture of benzyl alcohol (100 mL) and benzene (250 mL). • The mixture was heated for 24 hours using a Dean Stark trap. The reaction mixture was concentrated and diethyl ether was added to precipitate a solid. The solid was filtered, washed with diethyl ether and dried to give L-4-trans-hydroxyproline benzyl ester (75 g).

(2) To a suspension of the L-4-trans-hydroxyproline benzyl ester p-toluenesulfonate (40.43 g) obtained above in tetrahydrofuran (500 mL) and DIEA (51.3 mL), 3,5 -Dichlorophenyl isocyanate (22.1 g) was added. After stirring overnight, the reaction mixture was concentrated. The residue was dissolved in ethyl acetate, washed, dried, filtered and concentrated. The residue was triturated in ethyl acetate / hexane (1: 1), the white solid was filtered and purified by flash column chromatography (silica gel; hexane / ethyl acetate = 2: 1) (2S, 4R) -2. -[(3,5-dichlorophenyl) carbamoyl] -4-hydroxyproline benzyl ester was obtained (33.07 g).

(3) To a suspension of (2S, 4R) -2-[(3,5-dichlorophenyl) carbamoyl] -4-hydroxyproline benzyl ester (33.07 g) in acetonitrile (800 mL), imidazole (11 g) and tert-Butyldimethylsilyl chloride (13.64 g) was added. After stirring for 48 hours, the reaction mixture was concentrated. The residue was dissolved in ethyl acetate, washed, dried, filtered and concentrated. The residue was purified by flash column chromatography (silica gel; hexane / ethyl acetate = 2: 1) to give (2S, 4R) -2-[(3,5-dichlorophenyl) carbamoyl] -4- (tert-butyldimethylsilyloxy) ) Proline benzyl ester was obtained (44.45 g).

(4) To a solution of (2S, 4R) -2-[(3,5-dichlorophenyl) carbamoyl] -4- (tert-butyldimethylsilyloxy) proline benzyl ester (23.49 g) in acetonitrile (500 mL), DIEA ( 34.44 mL) was added and the mixture was heated to reflux. After refluxing for 24 hours, the reaction mixture was concentrated and purified by flash column chromatography (silica gel; hexane to hexane / ethyl acetate = 1: 1) to give 3- (3,5-dichlorophenyl) -7- (tert- The two diastereomers of butyldimethylsilyloxy) -1,3-diazabicyclo [3.3.0] octane-2,4-dione were separated.
Diastereomer A: 7.46 g, MS: m / z 415 (M ⁺ ), and Diastereomer B: 10.66 g, MS: m / z 415 (M ⁺ ).

(5) The compound obtained above (diastereomer A or B: 12.73 g) was benzylated as follows. Under a nitrogen atmosphere, n-butyllithium (1.6 M hexane solution, 30 mL) was added to a tetrahydrofuran (100 mL) solution of diisopropylamine (6.5 mL) at −78 ° C. with stirring. The mixture was maintained at -78 ° C for an additional 30 minutes. Of 3- (3,5-dichlorophenyl) -7- (tert-butyldimethylsilyloxy) -1,3-diazabicyclo [3.3.0] octane-2,4-dione (12.73 g) under nitrogen atmosphere To the tetrahydrofuran (100 mL) solution was added the mixture at −78 ° C. using a cannula. After stirring at −78 ° C. for 30 minutes, a solution of 4-cyano-α-bromotoluene (9.08 g) in tetrahydrofuran (100 mL) was added. The reaction mixture was stirred at −78 ° C. for 2.5 hours, then allowed to warm slowly to room temperature and left at room temperature for 0.5 hour. The reaction mixture was concentrated and the residue was dissolved in ethyl acetate. The ethyl acetate solution was washed, dried, filtered and concentrated. The residue was purified by flash column chromatography (silica gel; hexane / ethyl acetate = 24: 1 to 3: 1) to give (5S, 7R)-and (5R, 7R) -5- (4-cyanobenzyl) -3- (3,5-Dichlorophenyl) -7- (tert-butyldimethylsilyloxy) -1,3-diazabicyclo [3.3.0] octane-2,4-dione was obtained.
(5S, 7R) isomer: 7.6 g, MS: m / z 530 (M ⁺ ) and (5R, 7R) isomer: 1.8 g, MS: m / z 530 (M ⁺ ).

(6) (5S, 7R) -5- (4-Cyanobenzyl) -3- (3,5-dichlorophenyl) -7- (tert-butyldimethylsilyloxy) -1,3-diazabicyclo [3.3.0 A solution of octane-2,4-dione (1.0 g) in tetrahydrofuran (1 mL) was added to 70% HF / pyridine (25 mL). The reaction mixture was stirred for 24 hours and then concentrated. The residue was dissolved in ethyl acetate and the resulting solution was washed, dried and concentrated. The residue was purified by flash column chromatography (silica gel; methanol / dichloromethane = 2-7%) to give (5S, 7R) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-hydroxy. -1,3-diazabicyclo [3.3.0] octane-2,4-dione (0.52 g) was obtained. MS (m / z) 416 [MH ^<+ >].

(7) (5S, 7R) -5- (4-Cyanobenzyl) -3- (3,5-dichlorophenyl) -7-hydroxy-1,3-diazabicyclo [3.3.0] octane- obtained above To a solution of 2,4-dione (0.52 g) in dichloromethane (8 mL) at 0 ° C. was added DIEA (0.45 mL) followed by methanesulfonyl chloride (0.15 mL) and the mixture was stirred for 1.5 hours. . The reaction mixture is diluted with dichloromethane and the resulting mixture is washed, dried, filtered and concentrated to (5S, 7R) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl)- 7-methanesulfonyloxy-1,3-diazabicyclo [3.3.0] octane-2,4-dione was obtained (0.76 g). This compound was used as such in the next step. MS (m / z) 501 [MH ^<+ >].

(8) (5S, 7R) -5- (4-Cyanobenzyl) -3- (3,5-dichlorophenyl) -7-methanesulfonyloxy-1,3-diazabicyclo [3.3.0] obtained above. Sodium azide was added to a solution of octane-2,4-dione (0.76 g) in dimethylformamide (5 mL) and the mixture was stirred for 24 hours. The reaction mixture was partitioned between ethyl acetate and water. The organic solution was washed, dried, filtered and concentrated. The residue was purified by flash column chromatography (silica gel; dichloromethane) to give (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-azido-1,3-diazabicyclo [ 3.3.0] Octane-2,4-dione (0.46 g) was obtained. MS (m / z) 441 [MH ^<+ >].

(9) (5S, 7S) -5- (4-Cyanobenzyl) -3- (3,5-dichlorophenyl) -7-azido-1,3-diazabicyclo [3.3.0] octane- obtained above Hydrogen was bubbled into a mixture of 2,4-dione (0.42 g), 5% palladium-carbon (15 mg) and ethanol (15 mL) and the reaction mixture was stirred overnight under a hydrogen atmosphere. The reaction mixture was filtered through celite, and the filtrate was concentrated. The residue was purified by HPLC (Beckman 5 μC18 column; gradient elution, water / acetonitrile (10-100%) / 0.1% trifluoroacetic acid) (5S, 7S) -5- (4-cyanobenzyl) -3- ( 3,5-Dichlorophenyl) -7-amino-1,3-diazabicyclo [3.3.0] octane-2,4-dione (0.21 g) was obtained. MS (m / z) 415 [MH ^<+ >].

Experimental Example 1 Cell Adhesion Inhibitory Action Recombinant ICAM-1 was constructed from fusion of 5 extracellular domains of human ICAM-1 and human IgG constant region. Recombinant ICAM-1 (hereinafter ICAM-1) was purified by protein A affinity chromatography and stored at −20 ° C. ICAM-1 was diluted with PBS (pH 7.5), transferred to a Falcon Probind III plate at 100 μl / well, and fixed by incubation at 4 ° C. overnight. BSA coated wells were used as a basis for non-specific background adhesion. The washed plate was blocked with 0.25% PBS solution of ovalbumin at 37 ° C. for 1 hour. HBSS washed Jurkat cells were adjusted to a final concentration of 2.5 × 10 ⁶ / mL with TBSg adhesion buffer (24 mM Tris, pH 7.4, 0.14 M sodium chloride, 2.7 mM potassium chloride, 2 mM glucose, 0.1% HSA). This suspension (100 μl) was added to an ICAM-1 coated plate containing 100 μl of plate buffer (containing TBSg buffer, 10 mM magnesium chloride, 2% dimethyl sulfoxide). Adhesion was performed at 37 ° C. for 1 hour. Nonadherent cells were removed using an EL404 plate washer (Biotech Instruments; Highland Park, Vermont). The number of adherent cells was quantified by measuring the enzyme activity of endogenous N-acetyl-hexosaminidase using p-nitrophenol-N-acetyl-β-D-gluco-saminide as the enzyme substrate. The amount of p-nitrophenol released was measured by reading the optical density at 405 nm with a vertical path spectrophotometer (VMAX Kinetic Microplate Reader, MOLECULAR DEVICES, Menlo Park, California) and cell adhesion Quantified. The test compound was dissolved in dimethyl sulfoxide, diluted to the required concentration with plate buffer, and then transferred to the ICAM-1 coated plate. The concentration (IC ₅₀ ) at which the test compound (compound described in the production example) inhibits adhesion (cell adhesion) between Jurkat cells and ICAM-1 coated plates by 50% was calculated. The results are shown in Table 1.

Experimental Example 2 Neurokinin 1 (NK1) Receptor Binding Inhibitory Effect According to the method described in European Journal of Pharmacology 254, 221-227 (1994), human lymphoblastoma Cells (IM-9) (4 × 10 ⁶ cells) were prepared at 0.3 nM [ ³ H] (Sar ⁹ , Met ¹¹ (O ₂ )) substance P (Kd value: 0.17 nM) and a common ratio of 10. 50 mM Tris-HCl (pH 7.4, 25 containing 150 mM sodium chloride, 3 mM manganese chloride, 40 μg / mL bacitracin, 4 μg / mL leupeptin, 4 μg / mL chymostatin, 4 μg / mL phosphoramidon, 0.02% bovine serum albumin together with the test compound. (C) at room temperature for 60 minutes in 0.5 mL. Suction-filter with a GF / C glass filter pretreated with 0.3% polyethyleneimine, wash twice with 3 mL of ice-cold reaction buffer without bovine serum albumin and various protease inhibitors, and filter with a liquid scintillation counter The upper radioactivity (dpm) was measured (total binding). The nonspecific binding amount was measured in the same manner using L-703606 (2 μM) having an NK1 receptor antagonistic action. The specific binding amount was obtained by subtracting the non-specific binding amount from the total binding amount, and the inhibition rate of the sample compound for the specific binding of the labeled ligand at each concentration was calculated to calculate the 50% inhibitory concentration (IC ₅₀ ). . The results are shown in Table 2. L-703606 is a compound represented by the following formula.

The active ingredient (I) of the present invention has an α _L β ₂ integrin-mediated cell adhesion inhibitory action and a substance P receptor antagonistic action. For this reason, the pharmaceutical composition of the present invention can be used as a therapeutic or prophylactic agent for a pathological condition associated with LFA-1 mediated cell adhesion, and as a therapeutic or prophylactic agent for a pathological condition caused by or mediated by substance P.

Claims (18)

Formula (I):

(Wherein R represents a hydrogen atom, a hydroxyl group or a carbamoyl group, and n represents 1 or 2)
Or a pharmacologically acceptable salt thereof as an active ingredient.   The pharmaceutical composition according to claim 1, wherein R is a hydrogen atom.   The pharmaceutical composition according to claim 1, wherein R is a hydroxyl group.   The pharmaceutical composition according to claim 1, wherein R is a carbamoyl group.   The pharmaceutical composition according to any one of claims 1 to 4, wherein n is 1.   The pharmaceutical composition according to any one of claims 1 to 4, wherein n is 2.   The pharmaceutical composition according to claim 1, wherein R is a hydrogen atom and n is 1.   The pharmaceutical composition according to claim 1, wherein R is a hydroxyl group and n is 1.   The pharmaceutical composition according to claim 1, wherein R is a carbamoyl group and n is 2. The compound is (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-acetylamino-1,3-diazabicyclo [3.3.0] octane-2,4 -Dione,
(5S, 7S) -5- (4-Cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(2-hydroxyacetyl) amino] -1,3-diazabicyclo [3.3.0] octane -2,4-dione and (5S, 7S) -5- (4-cyanobenzyl) -3- (3,5-dichlorophenyl) -7-[(3-carbamoylpropionyl) amino] -1,3-diazabicyclo [3.3.0] The pharmaceutical composition according to claim 1, selected from octane-2,4-dione.   The pharmaceutical composition according to any one of claims 1 to 10, which is a therapeutic or prophylactic agent for LFA-1 mediated disease states.   The pharmaceutical composition according to claim 11, wherein the LFA-1-mediated condition is an inflammatory disease, an autoimmune disease or an allergic disease.   The pharmaceutical composition according to any one of claims 1 to 10, which is an agent for treating or preventing a disease caused by or mediated by substance P.   The pharmaceutical composition according to claim 13, wherein the disease is an inflammatory disease. Rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, allergic disease, adult respiratory distress syndrome, AIDS, cardiovascular disease, thrombosis, harmful platelet aggregation, reocclusion after thrombolysis, reperfusion injury, skin inflammatory disease, osteoporosis, Osteoarthritis, atherosclerosis, arteriosclerosis, neoplastic disease, trauma, retinal detachment, type I diabetes, multiple sclerosis, systemic lupus erythematosus, ocular inflammatory condition, inflammatory bowel disease, cystitis, stomach The pharmaceutical composition according to any one of claims 1 to 10, which is a therapeutic or prophylactic agent for a disease selected from a disorder, localized ileitis, Sjogren's syndrome, and rejection after organ transplantation.   16. The pharmaceutical composition according to claim 15, wherein the disease is psoriasis, rheumatoid arthritis, inflammatory bowel disease, systemic lupus erythematosus, atopic dermatitis, Sjogren's syndrome or rejection after organ transplantation.   16. The pharmaceutical composition according to claim 15, wherein the disease is rheumatoid arthritis, asthma, chronic obstructive pulmonary disease, psoriasis, multiple sclerosis or rejection after organ transplantation. The pharmaceutical composition according to claim 15, wherein the disease is asthma, inflammatory bowel disease, cystitis or gastric disorder.