NZ620469B2 - Pyrrolidine-3-ylacetic acid derivative - Google Patents

Abstract

Provided are pyrrolidine-3-yl-acetic acid derivative compounds, of the general formula (1), wherein the variables are as defined in the specification. Examples of the compounds include 2-[(3S,4R)-1-{[2-chloro-6-(trifluoromethyl)phenyl]methyl}-3-{[1-(2-fluoropentyl)piperidin-4-yl]carbamoyl}-4-methylpyrrolidin-3-yl]acetic acid and 2-[(3S,4R)-3-{[(3S)-1-(cyclohex-1-en-1-ylmethyl)pyrrolidin-3-yl]carbamoyl}-1-[(2,6-dichlorophenyl)methyl]-4-methylpyrrolidin-3-yl]acetic acid. The compounds are inhibitors of the fractalkine-CX3CR1 pathway. The compounds may be useful in the treatment of inflammatory bowel diseases, such as ulcerative colitis and Crohn’s disease. yrrolidin-3-yl]acetic acid and 2-[(3S,4R)-3-{[(3S)-1-(cyclohex-1-en-1-ylmethyl)pyrrolidin-3-yl]carbamoyl}-1-[(2,6-dichlorophenyl)methyl]-4-methylpyrrolidin-3-yl]acetic acid. The compounds are inhibitors of the fractalkine-CX3CR1 pathway. The compounds may be useful in the treatment of inflammatory bowel diseases, such as ulcerative colitis and Crohn’s disease.

Description

FP11-0417 DESCRIPTION Title ofInvention PYRROLIDlNEYLACETICACID DERIVATIVE cal Field The present invention relates to a pyrrolidin—3-ylacetic acid derivative. More particularly, the present invention relates to a pyrrolidin—3-ylacetic acid derivative having availability as a therapeutic agent for inflammatory bowel disease.

BackgroundArt Chemokines are major cell migration factors and regulate infiltration of 1’0 lymphOcytes into tissues through the ement of cell nt and the tion of adhesion molecules. Chemokines are classified into four subfamilies of CC, CXC, C and CX3C based on their sequences ofthe first two ne residues.

Fractalkine is the sole CX3C chemokine member and has distinct characteristics in its structure and fimctions which are not found in other Chemokines. Fractalkine binds to a receptor, CX3CR1, which can mediate strong adhesion without mediation of selectin or integrin even in the presence of a physiological blood flow. This means that the fiactalkine— CX3CR1 system mediates multi-stage infiltration mechanism through selectin or integrin by only a one—stage reaction. sion of fiactalkine on vascular endothelial cells is induced by inflammatory cytokines TNF and IL-1. On the other hand, CX3CR1 is expressed on monocytes, almost all NK cells and some T cells, but is not expressed on neutrophils. Therefore, the fiactalkine— CX3CR1 system is considered to be an extremely effective mechanism to mobilize immune cells onto the endothelial cells ofdamaged tissues or into the tissues.

\Vith regard to the relation between the fiactalldne—CXBCRI system and “pathologies, it is suggested that the fiactalkine—CXBCRI system is involved in the development and pathologies of autoimmune diseases such as rheumatoid arthritis, atory bowel disease, lupus tis and multiple sclerosis (Non Patent Literature 1).

In ular, with regard to inflammatory bowel disease, it is reported that expression of fractalkine is enhanced at inflammatory sites of colonic tissues ents and that CX3CR1 plays an important role in the infiltration of immune cells into the colon tissue (Non Patent ture 2).

Antibodies bed in Patent Literature 1 and low molecular weight nds described in Patent Literatures 2 to 6 have been previously known as fractalldne inhibitors.

FP11—0417 In addition, compounds described in Patent Literature 7 are described to be useful as ine CCR2 receptor nists, but dilfer in the target chemokine family from such inhibitors.

Citation List Patent Literature Patent Literature 1 : Japanese Patent Application Laid-Open Publication No. 2002-345454 Patent Literature 2: Patent Literature 3: Patent Literature 4: Patent Literature 5: Patent Literature 6: WC 2009/120140 Patent Literature 7: US. Patent Application Laid~0pen PublicationNo. 2010/0210633 Non Patent Literature

[0009] Non Patent Literature 1: Umehara et a1., "Fractalkine in Vascular Biology", Arterioscler.

Thromb. Vasc. Biol, Vol. 24, pp. 34—40, 2004 Non Patent ture 2: Kobayashi et al., "Exclusive Increase of CX3CR1_CD28_CD4_ T Cells in Inflammatory Bowel Disease and Their Recruitment as Intraepithelial Lymphocytes", Inflamm. Bowel. Dis, Vol. 13, pp. 837—846, 2007 y ofInvention Technical Problem An object of the present invention is to provide a nd having an tory efi‘ect in the fractalkine—CX3CR1 pathway.

Solution to Problem As a result of intensive studies, the present inventors have found the present invention Specifically, the present ion relates to A compound represented by formula (1) or pharmaceutically acceptable salt [Chemical Formula 1] FP11-0417 wherein R represents a C16 alkyl group unsubstituted or having 1 to 3 substituents selected from Substituent Group A, a C33 lkyl group unsubstituted or having 1 to 3 substituents ed from Substituent Group A, or a C33 cycloalkenyl group unsubstituted or having 1 to 3 substituents selectedfiom SubstituentG’roup A, X represents a C16 alkyl group, Y and Z are the same or nt from each other and each represents a halogen atom or a C16 alkyl group unsubstituted or having 1 to 3 substituents selected from Substituent Group n represents 0 or 1, Substituent GroupA consists ofhalogen atoms, and Substituent Group B consists ofhalogen atoms; The nd or pharmaceutically acceptable salt thereof according to [1], wherein R is a fluorobutyl group, a pentyl group, a cyclohexyl group, a difluorocyclohexyl group, a cyclopentenyl group or a cyclohexenyl group; The compound or pharmaceutically acceptable salt thereof according to [1] or [2], whereinX is amethyl group; The compound or pharmaceutically acceptable salt faccording to any one of to [3], wherein Y is a chlorine atom; [5] The compound or pharmaceutically acceptable salt thereofaccording to any one of to [4], wherein Z is a ne atom, a methyl group, a difluoromethyl group or a trifluoromethyl group; The compound or phannaceutically acceptable salt thereof according to any one of to [5], whereinnis l; [7] Acompound selected from the group consisting of: 2-[(3S,4R){[2-chloro(uifluoromethyl)phenyl]methyl}-3—{[1-(2-fluoropentyl)piperidin— 4-yl]carbamoyl}—4-mefl1ylpyrrolidin—3-yl]acetic acid, ,4R)—1-[(2,6-dichlorophenyl)methyl]-3—({ 1-[(4,4-difluorocyclohexyl)methyl]piperidin— FPll-0417 4-yl}carbamoyl)-4—methylpyrrolidin—3~yl]acetic acid, ,4R)-l-{[2-chloro(11ifluoromethyl)phenyl]methyl}{[l-(cyclohex-l-en-lylmethyl )piperidin—4—yl]carbamoyl}methylpyrrolidin—3~yl]acetic acid, 2—[(3S,4R)~l-[(2—chloromethylphenyl)methyl]{[l—(cycloheX-1—en—1- ylmethyl)piperidin—4—yl]carbamoyl}—4—methylpyrrolidin—3-y1]acetic acid, 2-[(3S,4R)—1—{[Z-chloro-6—(trifluoromethyl)phenyl]methyl}{[l-(cyclopent—l-en—l— yhnethyl)piperidin—4-yl]carbamoyl}—4—methylpyrrolidinyl]acetic acid, 2-[(3S,4R)[(2-chloromethylphenyl)methy1]—3-{[(1 -cyclopent—1-en—l- ylmethyl)piperidin—4—yl]carbamoyl}—4—methylpyrrolidin~3~yl]acetic acid, 2-[(3S,4R)—(3—{[(3S)—1~(cyclohex~l-en—l~ylmethyl)pyrrolidin~3~yl]carbamoyl}~1-[(2,6- dichlorophenyl)methy1]—4-methy1pyrrolidin—3-yl]acetic acid, 2—[(3S,4R)—l-{[2-chloro—6—(difluorometliyl)phenyl]mefl1yl}[(1—hexylpiperidin—4— bamoyl]-4—methylpyrrolidin—3—yl]acetic acid, 2—[(3S,4R){[(l -cyclohex-1~en-l~ylmethyl)pipexidin—4-yl]carbamoyl}-l{(2,6- dichlorophenyl)methyl]methylpyrrolidinyl]acetic acid, 2-[(3S,4R)—l-{[2-chloro(difluoromethyl)phenyl]methyl}{[1-(cyclohex—1-en—1- yl)piperidin—4-y1]carbamoyl}-4—methylpy1rolidinyl]acetic acid, 2—[(3S,4R)—l-{[2—chloro(difluoromethyl)phenyl]methyl}—3—{[l-(cyclopent—l—en—l— ylmethyl)piperidin—4—yl]carbamoyl}—4—methylpy1rolidin—3—yl]acetic acid and 2-[(3S,4R)-l—{[2-chloro-6{difluoromethprhenyflmethyl}{»[1— (cyclohexyhnethyl)piperidin—4-yl]carbamoyl}—4—methy1pyrrolidin—3-yl]acetic acid, or a pharmaceutically acceptable salt thereof; A medicine comprising the compound or pharmaceutically acceptable salt thereof according to any one of [l] to [7] as an active ient; [9] A therapeutic agent for inflammatory bowel disease comprising the compound or pharmaceutically acceptable salt thereof according to any one of [1] to [7] as an active ingredient; The therapeutic agent according to [9], wherein the inflammatory bowel disease is ulcerative colitis or Crohn's e; ' [l l] A kine—CX3CR1 pathway inhibitor comprising the nd or pharmaceutically acceptable salt thereof according to any one of [1] to [7] as an active ingredient; A fractalkine inhibitor comprising the compound or phannaceutically acceptable FP11-0417 salt thereofaccording to any one of [1] to [7] as an active ingredient; A CX3CR1 inhibitor sing the compound or pharmaceutically acceptable salt thereofaccording to any one of [1] to [7] as an active ingredient; A method for treating inflammatory bowel disease comprising administering the compound or pharmaceutically acceptable salt thereof according to any one of [1] to [7] to a patient; The method according to [14], wherein the inflammatory bowel disease is ulcerative s or Crohn's disease; A method for ting the fiactalldne—CDBCR1 pathway comprising administering the compound or pharrnaceutically able salt thereof according to any one of [1] to [7] to apatient; A method for inhibiting lkine comprising administering the compound or pharmaceutically acceptable salt thereofaccording to any one of [1] to [7] to a patient; A method for inhibiting CX3CR1 comprising administering the compound or phannaceutically acceptable salt thereofaccording to any one of [1] to [7] to a patient; The compound or aceutically acceptable salt thereof according to any one of to [7], which is used for the treatment ofinflammatory bowel disease; The compound or pharrnaceutically acceptable salt thereof according to [19], wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease;

[21] The compoundor pharmaceutically acceptable salt thereof according to any one of to [7], which is used for the inhibition ofthe lkine-CX3CR1 pathway; The compound or ceutically acceptable salt thereofaccording to any one of to [7], which is used for the inhibition offiactalkine; The compound or pharmaceutically able salt thereof according to any one of [1] to [7], which is used for the inhibition ofCX3CR1; Use of the compound or pharmaceutically acceptable salt thereof according to any one of [1] to [7] in the manufacture ofa therapeutic agent for inflammatory bowel disease; The use ing to [24], wherein the inflammatory bowel disease is tive colitis or Crohn's disease;

[26] Use ofthe nd or pharmaceutically acceptable salt thereof according to any one of [1] to [7] in the manufacture ofa fiactalldne-CX3CR1 pathway inhibitor; Use ofthe compound or pharrnaceuticaily acceptable salt thereof according to any one of [1] to [7] in the manufacture ofa lkine inhibitor; and FP11-0417 Use ofthe compound or phannaceutically acceptable salt thereof ing to any one of [1] to [7] in the manufacture ofa CX3CR1 tor.

Advantageous Effects ofInvention According to the results ofthe tests described below, the compounds according to the present invention have an inhibitory efiect in the fiactalldne—CX3CR1 pathway.

Therefore, the compounds according to the present invention have availability as therapeutic agents for inflammatory bowel e.

BriefDescription ofDrawings FIG 1 shows a graph showing the results of Test Example 2 for the compounds of Examples 3,6 and 11; FIG 2 shows a graph showing the results of Test Example 2 for the compounds of Examples 2, 7 and 8; FIG 3 shows a graph showing the results of Test e 2 for the compounds of es 1, 9 and 10; and FIG 4 shows a graph showing the results of Test Example 2 for the compounds of Examples 12, 13 and 14.

Description ofEmbodiments The present invention will be described in detail below.

In the present specification, the present invention is not limited to a particular crystal form but may include any one of l forms or mixtures thereof, although crystal polymorphs may exist. The present invention also includes amorphous forms, and the compounds according to the t invention include anhydrides, hydrates and solvates.

Hereinafter, the meanings of terms, symbols and the like described in the present specification will be described, and the present invention will be bed in detail.

[0017] The "CM; alkyl group" in the present cation means a linear or branched alkyl group having 1 to 6 carbon atoms, and examples include a methyl group, an ethyl group, a 1- propyl group, a 2—propy1 group, a 2-methyl—1-propyl group, a 2-methyl—2—propyl group, a 1- butyl group, a l group, a 1-pentyl group, a 2~penty1 group, a yl group, a 1-hexyl group, a l group and a 3-hexyl group.

[0018] The "C33 cycloalkyl group" in the present specification means a monocyclic saturated aliphatic hydrocarbon group having 3 to 8 carbon atoms, and examples include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group and a cyclooctyl group.

FP11-0417 The "ng cycloalkenyl group" in the t specification means a monocyclic aliphatic hydrocarbon group having 3 to 8 carbon atoms and containing 1 to 4 double bonds in the ring, and examples include a cyclopropenyl group, a cyclobutenyl group, a cyclopentenyl group, a exenyl group, a cycloheptenyl group and a cyclooctenyl group.

The "halogen atom" in the present specification means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

R in the compound represented by formula (1) represents a C16 alkyl group unsubstituted or having 1 to 3 substituents selected from Substituent Group A, a C33 cycloalkyl group unsubstituted or having 1 to 3 substituents selected from tuent Group A, or a C34; cycloalkenyl group unsubstituted or having 1 to 3 ments ed from Substituent Group A. Preferably, R represents a fluorobutyl group, a pentyl goup, a cyclohexyl group, a difluorocyclohexyl group, a entenyl group or a cyclohexenyl group.

X in the compound represented by formula (1) represents a C16 alkyl group.

Preferably, X represents a methyl group.

YandZ in the compound represented by formula (1) are the same or different from each other and each represents a halogen atom or a C16 alkyl group unsubstituted or having 1 to 3 substituents selected from Substituent Group B. Preferably, Y represents a chlorine atom. Preferably, Z represents a chlorine atom, a methyl group, a difluoromefliyl group or a trifluoromethyl group. n in the compound represented by formula (1) represents 0 or 1, and preferably ents 1.

Substituent GroupA consists ofhalogen atoms, and is preferably a fluorine atom.

Substituent Group B consists ofhalogen atoms, and is preferably a fluorine atom, a chlorine atom or a e atom, more ably a fluorine atom.

The "pharmaceutically acceptable sal " in the present specification is not particularly limited insofar as it forms a salt with the compound represented by formula (1) and is pharmaceutically acceptable, and es include inorganic acid salts, organic acid salts, inorganic base salts, c base salts, and acidic or basic amino acid salts.

[0028] Preferred examples of inorganic acid salts include hydrochlorides, hydrobromides, sulfates, nitrates and phosphates, and preferred examples of organic acid salts include acetates, ates, fumarates, maleates, tes, citrates, lactates, stearates, benzoates, mandelates, methanesulfonates, ethanesulfonates, p—toluenesulfonates and FP11-0417 benzenesulfonates.

Preferred es ofinorganic base salts include alkali metal salts such as sodium salts and potassium salts, ne earth metal salts such as calcium salts and ium salts, aluminum salts and ammonium salts, and preferred examples of organic base salts include diethylamine salts, diethanolamine salts, meglumine salts and N,N'— dibenzylethylenediamine salts.

Preferred examples of acidic amino acid salts include aspartates and glutamates, and red examples of basic amino acid salts include arginine salts, lysine salts and ornithine salts.

[0031] The compound represented by formula (1) can be produced by the method described below, and can also be produced by an improvement of the method described below by those skilled in the art based on the common knowledge. However, the method for producing the compound represented by formula (1) is not limited to these methods.

The compound represented by formula (1) (hereinafter also ed to as Compound (1)) can be produced through Process A, Process B and Process C described below in detail using an intermediate represented by formula (2) as a starting material. cal Formula 2] Process A Process B Process C [In the scheme, R, X, Y, Z and n are as defined above]

[0033] The sequence ofthe respective ses may be changed as appropriate based on the common knowledge of those skilled in the art. Each process may be followed by purification methods known to the d in the art, or may progress to next process without isolation and ation.

(Process A) Amidation [Chemical Formula 3] FP11-0417 Process A 0 ) R o Xé /0 n qu/< ""< HN N—\ N )n (2) (4) in X, R and n are as defined above] This process is a process of producing “Compound (4) by subjecting the carboxyl group ofCompound (2) and the amino group ofCompound (3) to dehydration condensation in an inert solvent in the ce ofa condensing agent and a base to form an amide bond.

The solvent used is not ularly limited insofar as it dissolves the starting material to some degree and does not inhibit the reaction, and examples include amides such as N,N-dimethylfonnamide, N,N—dimethy1acetamide and l-methylpyrrolidinone, ethers such as teuahydrofinan, and sulfoxides such as dimethyl sulfoxide, with methylfonnamide being preferred.

Examples of the condensing agent used include benzotriazol—l- yloxyuis(pyrrolidino)phosphonium hexafluorophosphate (hereinafter called PyBOP), bis(2- oxo-3—oxazolidinyl)phosphinic chloride (hereinafter called BOP-Cl), benzotn'azol—l- yloxytris(dimethylamino)phosphonium hexafluorophosphate, 2-(benzottiazol—1—yl)—1,1,3,3- tetramethyluronium hexafluorophosphate and diethyl cyanophosphate, with PyBOP or BOP— Cl being preferred and PyBOP being most preferred.

Examples of the base used include triethylamine and diisopropylethylamine, with triethylamine being preferred.

The reaction temperature varies depending on the starting material, solvent, condensing agent and base, but is usually ~20°C to 100°C, preferably 0°C to 60°C.

The reaction time varies depending on the starting material, solvent, condensing agent and base, but is y 30 s to five days, preferably one hour to three days.

(Process B) thylation [Chemical Formula 4] FP11-0417 Process B [wherein R, X, Y, Z and n are as defined above, and W1 represents a halogen atom, an alkylsulfonyloxy group or an arylsulfonyloxy group] This process is a process of ing Compound (6) by reacting Compound (4) with Compound (5) in an inert t in the presence ofa base.

The solvent used is not particularly limited insofar as it dissolves the starting material to some degree and does not inhibit the reaction, and examples include amides such as N,N—dimethylfonnamide, methylacetamide and l—methylpyrrolidinone, and sulfoxides such as dimethyl sulfoxide, with methylformamide being preferred.

[0044] Examples of the base used include inorganic bases such as sodium carbonate and potassium carbonate, with potassium carbonate being preferred.

The reaction temperature varies depending on the starting al, solvent and base, but is usually -20°C to 100°C, preferably 0°C to 60°C.

The reaction time varies depending on the starting material, solvent and base, but is usually 30 minutes to five days, preferably 1 to 24 hours.

(Process C) Elimination —butyl group [Chemical Formula 5] Process C (6) CC (1) [wherein R, X, Y, Z and n are as defined above] FP11-0417 This process is a process of ing Compound (1) by reacting Compound (6) with an acid in the absence ofa solvent or in an inert solvent.

The solvent used is not particularly limited insofar as it dissolves the ng material to some degree and does not inhibit the reaction, and examples include halogenated hydrocarbons such as dichloromethane and chloroform, toluene, dioxane, water and a mixed solvent ofdioxane and water, with dichloromethane being preferred.

Examples ofthe acid used include carboxylic acids such as trifluoroacetic acid, and inorganic acids such as hydrochloric acid, with trifluoroacetic acid being red.

The reaction temperature varies depending on the starting material, solvent and acid, but is usually —20°C to 100°C, preferably 0°C to 40°C.

The reaction time varies depending on the starting material, solvent and acid, but is usually 30 s to one day, preferably 1 to 12 hours.

A production method through Process D and Process F may also be used as another method for producing Compound (1).

[0054] (Process D) Reductive amination [Chemical Formula 6] in X, Y and Z are as defined above] This process is a process of producing Compound (8) by reacting Compound (2) with Compound (7) and a reducing agent in an inert solvent in the presence or absence of an acid.

The solvent used is not ularly limited insofar as it dissolves the starting material to some degree and does not inhibit the reaction, and examples include ethers such as tetrahydrofilran, and alcohols such as methanol and ethanol, with tetrahydrofiiran or methanol being preferred.

Examples of the reducing agent used include borohydride compounds such as sodium triacetoxyborohydride, sodium cyanoborohydride and sodium borohydride, with sodium triacetoxyborohydride being preferred.

The acid may or may not be used in this process, and if used, the acid used is not particularly limited insofar as it does not inhibit the reaction, and is preferably acetic acid.

The on temperature varies depending on the starting material, solvent, reducing agent and acid, but is usually —20°C to 100°C, preferably 0°C to 60°C.

The on time varies ing on the starting material, t, ng agent and acid, but is usually 30 minutes to five days, preferably 1 to 48‘hours.

Even when Compound (2) being the starting material forms a salt, the reaction can be allowed to proceed by adding an organic amine such as triethylarnine in an amount ofone or more lents to the amount ofthe carboxylic acid fonning the salt.

(Process E) Hydrogenation Compound (4) in which the arylmethyl group is eliminated can be obtained by subjecting Compound (6) obtained in the above Process B to hydrogenation reaction Compound (4) can further be converted by Process B to Compound (6) to which an arylmethyl group having a difiemnt substituent is added.

[Chemical Formula 7] ; Process 8 :;Q>n_\RProcess E X Wig?Xc. .../ HN N ”M:N:QN1R~——> HZ‘QL‘”NRN N Z1 22 [wherein R, Z and n are as defined above, Y1 and Y2 have the same meaning as in Y above, and Z1 and 22 have the same meaning as in Z above] This process is a process of producing Compound (4) by reacting Compound (6) with hydrogen in an inert solvent in the presence of a ion catalyst to remove the arylmethyl group.

The solvent used is not ularly limited insofar as it dissolves the starting material to some degree and does not inhibit the reaction, and examples include alcohols such as methanol and ethanol, ethers such as tetrahydrofiiran, dioxane and dimethoxyethane, FP11—0417 and organic acid esters such as ethyl acetate, with ethers, alcohols, organic acid esters or mixed solvents thereofbeing preferred and methanol or ethanol being most red.

Examples ofthe reduction catalyst used include Pd/C, palladium hydroxide, Raney nickel, platinum oxide and um black, with Pd/C or ium hydroxide being red.

The on temperature varies depending on the starting material and solvent, but is usually 0°C to 70°C, preferably 10°C to 50°C.

The reaction time varies depending on the starting material, solvent and reaction temperature, but is usually 30 minutes to five days, preferably one to three days.

[0068] The hydrogen pressure during the reaction in the case of using the reduction catalyst is usually 0.5 to 10 atm, preferably 1 to 5 atrn.

Typically, the nd obtained in this process can be used in the next process only by filtering 011”the catalyst.

(Process F) Compound (1) can also be obtained by Process F.

[Chemical Formula 8] Process F (F4) (12) [wherein R, X Y, Z and n are as defined above] (Process F—1)Amidation FP11-0417 This process is a s ofproducing Compound (11) by subjecting the carboxyl group of nd (8) and the amino group of Compound (10) to dehydration condensation and can be performed by a method similar to that ess A.

(Process F-2) Elimination oftert—butyl group and tert—butyloxycarbonyl group This process is a process ofproducing nd (12) by reacting Compound (11) with an acid and can be med by a method similar to that ofProcess C.

(Process F-3) Reductive amination This process is a process ofproducing Compound (1) by reacting Compound (12) with aldehyde compound (13) in the presence ofa reducing agent and can be performed by a method similar to that ofProcess D.

Compounds (10) and (13) can be available as commercially available compounds, or can be readily produced fiom commercially available compounds by methods usually performed by those skilled in the art (Process G) Compound (2) can also be ed by Process G below.

[Chemical Formula 9] FP11—0417 Process G W2/\©\ (15) 0 ¥0 0 X 0 racemic-(Z) (2) [wherein X is as defined above, V ents a hydrogen atom or a methoxy group, and W2 ents a halogen atom] (Process G—l) Esterification This process is a process ofproducing Compound (16) by reacting Compound (14) with Compound (15) in an inert solvent in the presence of a base. This process can be performed according to s B.

(Process G—2) Cycloaddition This process is a process ofproducing Compound (17) by reacting Compound (16) with hoxymethyl)—N—(11irnethylsilylmethyl)benzylamine in an inert solvent in the presence ofan acid The solvent used is not particularly limited insofar as it dissolves the starting FP11—0417 material to some degree and does not inhibit the on, and examples include ethers such as ydrofinan, halogenated hydrocarbons such as dichloromethane and chloroform, and ic hydrocarbons such as benzene and toluene, with dichloromethane and toluene and a mixed solvent thereofbeing preferred.

The acid used may be any acid usually used by those skilled in the art and is preferably roacetic acid.

(The reaction temperature varies depending on the starting material, solvent, reducing agent and acid, but is usually ~20°C to 60°C, preferably 10°C to 40°C.

The reaction time varies ing on the starting material, solvent, reducing agent and acid, but is usually 30 minutes to five days, preferably 1 to 24 hours.

The reaction may be exothermic, it is preferable to add N—(methoxymethyl)-N— (trimethylsilyhnethyl)benzylamine dropwise While being careful about heat generation, after nd (16), t and acid are mixed.

(Process G—3) Alkylation This process is a process ofproducing Compound (1 8) by causing a base to act on Compound (17) in an inert solvent and then ng with tert—butyl bromoacetate.

The solvent used is not particularly limited insofar as it dissolves the starting material to some degree and does not inhibit the reaction, and examples include ethers such as tetrahydrofuran and diethyl ether, aliphatic hydrocarbons such as , aromatic 2O hydrocarbons such as toluene, and mixed solvents thereof, with tetrahydrofinan and a mixed solvent oftetrahydrofiiran and hexane being preferred.

Preferred examples ofthe base used include lithium salts oforganic amines such as lithium diisopropylamide and lithium bis(trimethylsily1)arnide, with lithium diisopropylamide and m bis(trimethylsilyl)amide being more preferred.

[0086] The on temperature varies depending on the starting material, solvent and base, but is usually -100°C to 50°C, ably —80°C to 40°C, most preferably -80°C to — 70°C.

The reaction time varies depending on the starting material, t and base, but is usually 30 minutes to five days, preferably 1 to 24 hours, most preferably 2 to 5 hours.

[0088] (Process G—4) Hydrogenation reaction This process is a process of reacting Compound (18) with hydrogen in an inert solvent in the presence ofa reduction catalyst to remove the benzyl group.

This process can be performed according to Process E above.

FP11-0417 (Process G—5) Chiral resolution This process is a process of obtaining Compound (2) by subjecting a racemate of Compound (2) to chiral resolution.

The solvent used for the mobile phase or sample charge in the resolution is not particularly limited insofar as it dissolves the starting material to some degree and does not have adverse efiecw on columns or samples, and examples include water, aqueous saline solutions, alcohols such as methanol, ethanol and 2-propanol, hexane, acetonitrile, tetrahydrofiiran, trifluoroacetic acid, diethylarnine, or mixed solvents thereof, with acetonitrile, ethanol, or a mixed solvent nol and hexane being red.

[0092] Examples of the column used for the resolution include various commercially ble columns for optical resolution, with CHIRALPAK AD—H, CHIRALPAK IA and CHIRALCELOZ-H ctured by Daicel Chemical ries, Ltd. being preferred.

The column temperature during the resolution is preferably 10°C to 45°C.

(Process H) Compound (3) used in s A or the like can also be produced by Process H below.

[Chemical Formula 10] Process H .<_ o (:R ~<O- o 0—5N m HZNCF}? “‘4' NH m” ‘\ in H4 )n R H—2 n (19) (20) [wherein R and n are as defined above]

[0095] (Process H—l) Reductive ion This process is a process ofproducing Compound (20) by reacting Compound (19) with Compound (1 3) in the presence ofa ng agent in an inert solvent in the presence or absence ofan acid and can be performed according to Process D.

(Process H—2) Elimination oftert—butyloxycarbonyl group This process is a process ofproducing Compound (3) by ng Compound (20) with an acid in the absence of a solvent or in an inert solvent to remove the tert- butyloxycarbonyl group and can be performed according to Process C.

(Process 1) FP11-0417 Compound (2) can also be produced by Process I below using a te of Compound (2) as a starting material.

[Chemical Formula 11] Process I o >40 yo V o 0 X 0 x o X 0 l 1 OH N ii 0A0 ©Ao/ko racemic-(Z) V (21) (22) chiral—(22) inX andV are as defined above] (Process I—l) Introduction ofbenzyloxycarbonyl group This process is a process of reacting a racemate of Compound (2) with benzyl chlorofonnate in an inert t in the presence of a base to uce a benzyloxycarbonyl group.

[0099] The solvent used is not particularly limited insofar as it dissolves the starting material to some degree and does not inhibit the reaction, and es include ethers such as tetrahydrofiiran and 1,4-dioxane, water, N,N—dimethylformamide, dichloromethane, acetone and mixed solvents thereof, with a mixed solvent of water and acetone being preferred.

[0100] Examples of the base used include inorganic bases such as sodium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate and potassium carbonate, and c amines such as methylamine and diisopropylethylamine, with sodium hydroxide being preferred.

The reaction temperature varies depending on the starting al, solvent and base, but is usually -30°C to 20°C, preferably -10°C to 15°C.

FP11-0417 The reaction time varies depending on the starting material, t and base, but is usually 30 minutes to five days, preferably 1 to 24 hours.

(Process I-2) Esterification reaction This process is a process ofproducing Compound (22) by reacting Compound (21) with benzyl halide in an inert t in the ce of a base. This process can be performed according to Process B.

‘ (Process 1—3) Chiral resolution This process is a s of obtaining a chiral form of Compound (22) by subjecting Compound (22) to chiral resolution. This process can be performed according to Process GS.

(Process 1-4) Hydrogenation reaction This s is a process ofobtaining Compound (2) by reacting the chiral form of Compound (22) with hydrogen in an inert solvent in the presence of a reduction catalyst.

This process can be performed according to Process E.

[0106] (Process J) nd (2) can also be produced by Process J below using Compound (1 8) as a starting material.

[Chemical Formula 12] >40 >1o >4o 0 0 0 x 0 x 0 M +2 xfl ' ...../<0 o ____. o o N N N (>2 2 \> ©Ao’go 2 \> ©Ao’go 2 \> V V V chiral—(22) (18) (22) [whereinX andV have the same meaning as above] (Process J- 1) Introduction ofbenzyloxycarbonyl group FPll-0417 This process is a process ining Compound (22) by reacting Compound (18) with benzyl chloroformate in an inert solvent to replace the benzyl group with a benzyloxycarbonyl group.

The solvent used is not particularly limited r as it dissolves the starting material to some degree and does not inhibit the reaction, and examples include tetrahydrofuran, toluene, dichloromethane, chloroform and mixed solvents thereof, with dichloromethane being preferred.

The reaction temperature varies depending on the starting material and solvent, but is usually ~20°C to 60°C, preferably 0°C to 40°C.

[0110] The reactiOn time varies depending on the starting material and solvent, but is usually 30 minutes to five days, preferably 1 to 24 hours.

(Process J—2) Chiral resolution This process is a process of obtaining a chiral form of Compound (22) by subjecting Compound (22) to chiral resolution. This s can be performed according to Process G—S.

(Process J—3) Hydrogenation reaction This process is a process ofobtaining Compound (2) by reacting the chiral form of nd (22) with hydrogen in an inert solvent in the presence of a reduction catalyst.

This process can be performed according to Process E.

[0113] After completion of the reaction in each process of each method described above, the target compound in each process can be collected fiom the reaction e according to a tional method.

For e, when the Whole reaction mixture is a liquid, the reaction mixture is cooled to room temperature or cooled with ice as desired, and neutralized with an acid, alkali, ing agent or reducing agent as appropriate, an organic solvent immiscible with water and not ve with the target nd such as ethyl acetate is added, and the layer containing the target compound is separated. Next, a solvent immiscible with the resulting layer and not reactive with the target compound is added, the layer containing the target compound is washed, and the layer is separated. Moreover, when the layer is an organic layer, the target compound can be ted by drying with a drying agent such as anhydrous magnesium sulfate or ous sodium sulfate and ling ofi" the solvent. When the layer is an aqueous layer, the target compound can be collected by electrically demineralizing and then lyophilizing the layer.

FP11-0417 In addition, when the whole reaction mixture is a liquid and if le, the target compound can be collected only by distilling off substances other than the target nd (such as a solvent or a reagent) under normal pressure or reduced pressure.

Further, when only the target compound is precipitated as a solid, or when the whole reaction mixture described above is a liquid and only the target nd is precipitated in the course of collection, the target compound can be further ted by collecting the target compound by filtration first, washing the target compound collected by filtration with an appropriate c or inorganic solvent and drying, such that the mother liquor is treated in a manner similar to the case where the whole reaction mixture described above is‘a liquid.

Still r, when only the reagent or catalyst is present as a solid, or the whole reaction mixture described above is a liquid and only the reagent or catalyst is precipitated as a solid in the course of collection, and the target compound is dissolved in the solution, the target compound can be collected by filtering off the reagent or st first, washing the reagent or catalyst filtered offwith an appropriate organic or inorganic solvent, combining the resulting washings with the mother liquor, and ng the resulting mixture in a manner similar to the case where the whole reaction mixture described above is a liquid In particular, when substances other than the target compound which are contained in the reaction mixture do not inhibit the reaction in the next step, the reaction mixture may also be used in the next step as is Without particularly isolating the target nd.

Recrystallization, various chromatography methods and distillation may be carried out as riate in order to improve the purity of the target compound ted by the above method.

Typically, when the collected target compound is a solid, the purity of the target compound can be ed by recrystallization. In recrystallization, a single solvent or a mixture of a plurality of solvents not reactive with the target compound may be used Specifically, the target compound is first dissolved in one or more solvents not reactive with the target compound at room temperature or under heating. The resulting mixture is cooled with ice water or the like or is stirred or left to stand at room temperature, such that the target compound can be crystallized fiom the mixture.

The purity of the collected target compound can be ed by various chromatography methods. Generally, it is possible to use weak acidic silica gels such as Silica gel 60 manufactured by Merck KGaA (70-230 mesh or 340—400 mesh) and BW—300 FP11-0417 manufactured by Fuji Silysia Chemical Ltd. (300 mesh). When the target compound is basic and is adsorbed onto the above silica gels too strongly, it is also possible to use NH silica gels such as propylarnine coated silica gel manufactured by Fuji Silysia Chemical Ltd. (200—350 mesh) and disposable medium pressure preparative packed column manufactured by Yamazen Corporation (Hi—Flash Amino). When the target compound is dipolar or must be eluted with a more polar solvent such as methanol, for example, it is also possible to use NAM-200H or NAM-300H manufactured by NAM Laboratory or YMC "GEL ODS—A manufactured by YMC Co. Ltd. It is also possible to use disposable medium pressure preparative packed columns as bed above that are previously packed with fillers and manufactured by Yamaze’n 'CorpOration, Wako Pure Chemical Industries, Ltd., Biotage AB or W. R Grace & Co. (Hi-Flash Amino). The target compound whose purity is improved can be obtained by eluting the target compound with one or more solvents not ve with the target compound using these silica gels, and distilling offthe solvent(s).

When the ted target compound is a liquid, the purity ofthe target compound can also be improved by distillation. In distillation, the target compound can be distilled out by subjecting the target compound to reduced pressure at room ature or under heating.

Representative es of the method for producing Compound (1) have been described above. Raw material compounds and various reagents in the production of Compound (1) may form salts or solvates such as hydrates, all vary depending on the ng al, the solvent used or the like, and are not particularly limited insofar as they do not inhibit the reaction. Also, the solvent used varies depending on the ng al, the reagent or the like, and is not particularly limited insofar as it does not inhibit the reaction and dissolves the starting material to some degree, sly. When Compounds (1) are obtained as free forms, they can be converted to salts that may be formed by nds (1) or solvates ofthe compounds or salts by conventional methods.

When Compounds (1) are obtained as salts or solvates, they can be converted to free forms ofCompounds (1) by conventional s.

Various isomers obtained for Compounds (1) (such as geometric isomers, optical isomers, rotarners, stereoisomers and tautomers) can be purified and isolated using common 3O separation means, for example, recrystallization, reomeric salt ion, enzymatic resolution and various chromatography methods (such as thin layer tography, column chromatography and. gas chromatography).

Compounds (1) or pharmaceutically acceptable salts thereof can be formulated by FP11—0417 conventional methods, and examples of dosage forms include oral formulations (such as tablets, granules, powders, capsules and syrups), injections (for intravenous administration, intramuscular administration, subcutaneous administration and intraperitoneal administration) and external ations (such as tiansderrnal absorption formulations (such as ointments and patches), ophthalmic preparations, nasal ations and suppositories).

These solid formulations such as tablets, capsules, granules and powders may contain usually 0.001 to 99.5 wt%, preferably 0.01 to 90 wt% or the like, ofCompounds (1) or phannaceutically acceptable salts thereof.

When oral solid formulations are manufactured, tablets, granules, powders and capsules can be prepared by adding diluents, binders, disintegrants, lubricants, colorants or the like to compounds (1) or pharrnaceutically acceptable salts thereof as necessary and treating by tional methods. Tablets, granules, s, capsules and the like may also be film coated as necessary. es of diluents include lactose, corn starch and microcrystalline cellulose, examples ofbinders include hydroxypropylcellulose and hydroxypropylmethylcellulose, and examples of disintegrants e carboxymethylcellulose m and nnellose sodium.

Examples of lubricants include magnesium stearate and calcium stearate, and examples ofcolorants include titanium oxide.

[0131] Examples of film coating agents include hydroxypropylcellulose, ypropylmethylcellulose and cellulose.

Any excipients described above are not limited to these examples, obviously.

When injections (for intravenous administration, intramuscular administration, subcutaneous administration and intraperitoneal administration) are manufactured, they can be ctured by adding pH adjusters, buifers, suspending agents, solubilizing agents, antioxidants, preservatives (antiseptics), tonicity adjusting agents or the like to nds (1) or phannaceutically acceptable salts thereof as necessary and ng by conventional methods. Lyophilized formulations to be ved before use may also be prepared by lyophilization. These injections can be administered intravenously, subcutaneously and intramuscularly, for example. es of pH adjusters and buflem include c acids or inorganic acids and/or salts thereof, examples of suspending agents include methylcellulose, polysorbate 80 and carboxymethylcellulose sodium, examples of solubilizing agents include polysorbate 80 FP11—0417 and polyoxyethylene sorbitan monolaurate, examples of idants include or-tocopherol, examples of preservatives include methyl parahydroxybenzoate and ethyl droxybenzoate, and examples of tonicity adjusting agents include glucose, sodium chloride and tol; however, the excipients are not limited to these es, obviously.

These injections may contain usually 0.000001 to 99.5 wt%, preferably 0.00001 to 90 wt% or the like, ofCompounds (1) or pharrnaceutically acceptable salts thereof. "ii/When external formulations are manufactured, transdermal absorption ations (such as ointments and patches), eye drops, nasal drops, suppositories and the like can be manufactured by adding base materials and, as necessary, the fiers, preservatives, pH ers, colorants and the like described above to nds (1) or pharmaceutically acceptable salts thereof, and treating by conventional methods. s raw materials tionally used for pharmaceuticals, quasi drugs, ics and the like can be used as base materials, and examples include raw materials such as animal and vegetable oils, mineral oils, ester oils, waxes, higher alcohols and purified water.

These external formulations may contain usually 0.000001 to 99.5 wt%, preferably 0.00001 to 90 wt% or the like, of Compounds (1) or pharrnaceutically acceptable salts thereof.

The dosage of the medicine according to the present invention typically varies depending on the symptom, age, sex, weight or the like, but is acceptable if it is a dosage sufficient to produce a desired elfect. For example, for an adult, a dosage of about 0.1 to 5000 mg (preferably 0.5 to 1000 mg, more preferably 1 to 600 mg) per day is used in one dose during one or more days or in 2 to 6 divided doses for one day.

The present invention also include isotopically labeled Compounds (1), and such compounds are the same as Compounds (1), except that one or more atoms are replaced with an atom(s) having an atomic mass or mass number different fiom an atomic mass or mass number commonly found in nature. Isotopes that can be incorporated into Compounds (1) are es of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluoride, iodine and chlorine, for example, and include 2H, 3H, 11C, 14C, 13N, 15O, 18F, 32F, 35S, 123I and 1251.

[0141] nds (1) ning the above—described isotopes and/or other isotopes or pharmaceutically acceptable derivatives thereof (such as salts) fall within the claims of the present specification. The isotopically labeled compounds of the present invention, for example, compounds into which radioisotopes such as H and/or 14C are incorporated, are FP11—0417 useful for tissue distribution assays for medicines and/or substrates. 3H and 14C are considered to be useful because oftheir ease in preparation and detection. Isotopes 11C and 18F are considered to be usefiil for PET ron emission tomography), an isotope 1251 is considered to be useful for SPECT e photon emission computed aphy), and all these es are useful for brain imaging. Replacement with heavier isotopes such as 2H es certain therapeutic advantages such as an increase in the in vivo half-er due to higher metabolic stability, or a reduction in the required dose, and is therefore considered to be useful under certain circumstances. Isotopically labeled Compounds (1) can be uniformly prepared by performing the procedures disclosed in the following schemes and/or examples using readily available ically labeled reagents in place of non—"isotopica‘lly labeled reagents.

Compounds (1) can be used as chemical probes to trap target proteins in bioactive low molecular weight compounds. Specifically, Compound (1) can be converted to an affinity chromatography probe, a photoafiinity probe or the like by introducing a labeling group, a linker or the like into a moiety difiefing fiom a structural moiety essential for expression of activity of the compound by a que bed in J. Mass Spectrum. Soc.

Jpn, Vol. 51, No. 5, 2003, pp. 492-498 orW0 2007/139149 or the like.

Examples of labeling groups, linkers or the like used for chemical probes include groups shown in the group consisting of(l) to (5) below: (1) protein labeling groups such as photoaflinity labeling groups (such as a benzoyl group, a benzophenone group, an azido group, a carbonylazido group, a diaziridine group, an enone group, a diazo group and a nitro group) and chemical alfinity groups (such as a ketone group in which an oc-carbon atom is replaced with a halogen atom, a carbamoyl group, an ester group, an alkylthio group, Michael receptors such as or,B-unsaturated ketones and esters, and an oxirane group), (2) cleavable linkers such as -S-S-, O-, monosaccharides (such as a glucose group and a ose group) or disaccharides (such as e), and oligopeptide s cleavable by enzymatic reaction, (3) fishing tag groups such as biotin and a 3-(4,4-difluoro-5,7-dimethyl—4H—3a,4a—diaza—4— bora—s—indacen—3-yl)propionyl group, (4) detectable markers such as radiolabeling groups such as 1251, 32F, 3H and 14C; fluorescence labeling groups such as fluorescein, rhodamine, dansyl, umbelliferone, 7-nitrofinazanyl and a 3—(4,4-difluoro—5,7-dimethyl—4H—3a,4a-dia2a—4—bora—s—indacen—3-yl)propionyl group; FP11—0417 chemiluminescent groups such as luciferin and luminol; and heavy metal ions such as lanthanoid metal ions and radium ions; or (5) groups bound to solid phase carriers such as glass beads, glass beds, microfiter plates, agarose beads, agarose beds, polystyrene beads, polystyrene beds, nylon beads and nylon beds.

Probes prepared by introducing labeling groups or the like ed from the group consisting of (1) to (5) above into Compounds (1) according to the method desCribed in the above documents or the like can be used as chemical probes for identification of labeled proteins useful for searching for novel drug targets, for example. 1'0 Examples Compounds (1) can be produced by the s described in examples below, for example, and the effects of Compounds (1) can be confirmed by the methods described in test examples below. However, these methods are illustrative and the present invention is not limited to the following specific examples in any case.

[0145] When deuterium oxide is used as a measurement solvent in 1H—NMR for confirmation of the structures of compounds, the chemical shift of the spectrum of each compound is shown as a value corrected with the chemical shift ofthe solvent residual peak in deuterium oxide as 4.79.

(Example 1) Chiral form of ,4R)—1—{[2—chloro—6- (trifluoromethyl)phenyl]methyl}{[1-(2~fluoropentyl)piperidin—4~yl]carbamoyl}~4- methylpyrrolidin—3—yl]acetic acid (Example 1a) tert—Butyl 4—{[(benzyloxy)carbonyl]amino}piperidine—1—carboxy1ate tert—Butyl 4-aminopiperidine—1-carboxylate (10 g, 49.9 mmol), N,N- diisopropylethylamine (26 ml, 149 mmol), benzyl chloroformate (8.5 ml, 59.5 mmol) and dichloromethane (dehydrated) (300 ml) were mixed under ice—cooling. The resulting mixture was stirred at room temperature for one hour. An aqueous ted sodium bicarbonate on was added to the reaction mixture, which was extracted with dichloromethane three times. The c layer was washed with brine and dried over anhydrous sodium sulfate, and the solvent was distilled oif. The e was purified by silica gel column chromatography a gel, elution solvent: ethyl e/heptane) to give the title nd (13.1 g, yield 78.5%). 1H—NMR (400 MHz, CDC13) 5 ppm; .38 (2H, m), 1.45 (9H, s), 1.88-2.00 (2H, br), 2.80-2.94 (2H, br), 3.60-3.72 (1H, br), 3.92-4.10 (2H, br), 4.63-4.75 (1H, m), 5.09 (2H, s), FP11-0417 .40 (5H, m).

(Example lb) Benzyl N—(piperidin—4-yl)carbamate A mixture of tert-butyl 4-{[(benzyloxy)earbonyl]amino}piperidine—l-carboxylate obtained in e la (13.1 g, 39.2 mmol), a 5 N aqueous hydrochloric acid solution (40 ml, 200 mmol) and methanol (40 ml) was stirred at room temperature for 23 hours. A 5 N aqueous sodium hydroxide solution (40 ml) was added to the reaction mixture under ice- cooling. “Water and the solvent were distilled on from the reaction e While the mixture was azeotropically distilled with ethanol. l was added to the residue, the insoluble matter was removed by filtration, and the filtrate was concentrated to give the title compound (9.10 g, yield 99.1%). 1H—NMR (400 MHz, CDC13) 8 ppm; 1.31—1.42 (2H, m), 1.92—2.04 (2H, br), 2.70 (2H, d, J=12 Hz), 3.10 (2H, d, J=12 Hz), 3.56-3.68 (1H, br), 4.72—4.81 (1H, br), 5.09 (2H, s), 7.26- 7.40 (5H, m).

(Example 1c) Chiral form ofbenzyl N—[1-(2-fluoropentyl)piperidin—4-yl]carbamate (SS)-(-)—2,2,3-Trimethylbenzyl—4—imidazolidinone dichloroacetic acid (90 mg, 0.259 mmol), N—fluorobenzenesulfonimide (484 mg, 1.53 mmol), 2-propanol (0.4 ml) and tetrahydrofuran (dehydrated) (3.6 ml) were mixed at room temperature. The on mixture was cooled to —10°C, and pentanal (0.175 ml, 1.67 mmol) was then added, ed by stirring for three hours and 20 minutes while naturally warming from —10°C to room temperature. Benzyl N—(piperidin—4—yl)carbamate obtained in Example lb (304 mg, 1.3 mmol) and sodium triacetoxyborohydride (600 mg, 2.83 mmol) were added to the reaction mixture, which was stirred at room temperature for 15 hours and 40 minutes. Water and an aqueous sodium onate solution were added to the reaction mixture, the insoluble matter was removed by filtration, and the resulting e was extracted with ethyl acetate twice. The organic layer was washed with brine and dried over ous sodium sulfate, and the solvent was distilled ofi". The e was purified by silica gel column chromatography (silica gel, elution solvent: ethyl acetate/heptane) to give a chiral form mixture ofthe title compound (44 mg, yield 10.5%). 1H—NMR (400 MHZ, CDC13) 5 ppm; 0.94 (3H, t, J=7 Hz), .70 (6H, m), 1.85-2.00 (2H, br), 2.14-2.24 (2H, m), 2.35—2.48 (1H, m), 2.55—2.64 (1H, m), 2.80-3.00 (2H, m), 3.48-3.60 (1H, m), 4.59-4.72 (2H, m), 5.09 (2H, s), 7.26—7.37 (5H, m).

Optical resolution by HPLC (Analysis conditions) Column: CHlRALPAK AD-H (manufactured by Daicel Chemical FP11-0417 Industries, Ltd.) (0.46 cm diameter x 15 cm), eluent: hexane/ethanol = 9/1 (v/v), flow rate: 1 ml/mirr, detection: UV (210 nm) (Analysis result) The resulting chiral form mixture was analyzed under the above analysis conditions, and a peak with a retention time of 7.30 minutes (enantiomeric : 80% ee) and apeak with a retention time of 8.28 minutes were observed.

Further two lots of the chiral form mixture were ed by a method similar to the above method. Three lots in total ofthe chiral form mixture (451 mg, 1.4 mmol) were dissolved in ethanol (18 ml) and optically resolved repeatedly under the following onation conditions, and the peak with a shorter retention time was fiactionated to give the title compound (3 1 8 mg).

(Fractionation conditions) Column: CHIRALPAK AD-H (manufactured by Daicel Chemical ries, Ltd.) (2 cm er x 25 cm), eluent: hexane/ethanol = 9/1 (v/v), flow rate: 10 ml/min, detection: UV (220 nm) 1H—NMR (400 MHz, CDC13) 8 ppm; 0.94 (3H, t, J=7 Hz), 1.38-1.70 (6H, m), 1.85—2.00 (2H, br), 2.14-2.24 (2H, m), 2.35-2.48 (1H, m), 2.55-2.64 (1H, m), 2.80-3.00 (2H, m), 3.48—3.60 (1H, m), .72 (2H, m), 5.09 (2H, s), 7.26—7.37 (5H, m).

The resulting title compound was analyzed under the above analysis conditions to find that the retention time was 7.41 minutes and the enantiomeric excess was >99% ee.

(Example 1d) Chiral form of 1-(2-fluoropentyl)piperidin—4—amine A mixture of the chiral form (with a shorter retention time) of benzyl N—[l—(2— fluoropentyl)piperidin—4—yl]carbamate obtained in Example 10 (318 mg, 0.986 mmol), 10% Pd/C (100 mg) and methanol (7 ml) was stirred at room temperature for two hours under a hydrogen atmosphere. The atmosphere in the reaction vessel was replaced with nitrogen, % Pd/C was d ofli and the solvent was distilled off to give the title nd (249 mg). 1H—NMR (400 MHz, CDC13) 5 ppm; 0.95 (3H, t, J=7 Hz), 1.35-1.70 (4H, m), 1.92-2.10 (2H, br), 2.20-2.32 (2H, br), 2.48-2.62 (2H, br), 2.65-2.91 (2H, m), 3.18—3.30 (2H, br), 331—3 .42 (1H, br), 4.72—4.93 (1H, m).

(Example 1e) Benzyl (2E)-but—2-enoate Crotonic acid (70 g, 812 mmol) was dissolved in N,N—dimethylfom1amide (467 ml), which was cooled in an ice bath under nitrogen, and potassium carbonate (61.6 g, 447 mmol) was added. Benzyl bromide (91.7 ml, 772 mmol) was added dropwise to the reaction e over 20 minutes. The on mixture was stirred at room temperature for FPll-0417 18 hours. Ethyl acetate was added to the reaction mixture, which was filtered through Celite. The filtered ethyl acetate solution was washed with water, a saturated aqueous sodium bicarbonate solution and a saturated aqueous sodium de solution. The organic layer was dried over magnesium sulfate, filtered and concentrated under reduced pressure to give the title compound (142 g, yield: 99.4%). 1H—NMR (400 MHz, CDC13) 5 ppm; 1.87-1.90 (3H, m), 5.17 (2H, s), 5.87-5.92 (1H, m), 6.98-7.07 (1H, m), 7.26-7.39 (5H, m).

(Example If) Benzyl (3RS,4SR)~1—benzyl-4—methylpyrrolidinecarboxylate Benzyl ut—2—enoate obtained in Example le (20.5 g, 116 mmol) was ved in dichloromethane (5 ml), and the mixture was cooled in an ice bath with stirring.

Trifluoroacetic acid (257 til, 3.47 mmol) was added, and N~(methoxymethy1)-N— (trimethylsilylmethyl)benzylamine (33.1 g, 139 mmol) was added dropwise to the reaction liquid over 15 minutes so that the internal temperature did not exceed 62°C, while washing with dichloromethane (25 ml). The reaction liquid was left to stand until it reached room temperature and was stirred for 15 hours. The reaction liquid was concentrated and purified by silica gel column chromatography (elution solvent: ethyl acetate/heptane) to give the title compound (38 g). 1H—NMR (400 MHz, CDC13) 5 ppm; 1.14 (3H, d, J=6 Hz), 2.18-2.22 (1H, m), 2.48—2.65 (2H, m), 2.75—2.85 (2H, m), .94 (1H, m), 3.54-3.66 (2H, m), 5.13 (2H, s), 7.20—7.40 (10H, m).

(Example lg) Benzyl (3RS,4SR)benzyl—3—[2-(tert-butoxy)—2—oxoethyl]—4- methylpyrrolidine—3-carboxylate Benzyl (3RS,4SR)—1—benzy1—4—methylpyrrolidine~3—carboxy1ate obtained in Example 1f(30 g, 97.4 mmol) was dissolved in tetrahydrofuran (300 rrrl), which was cooled to -70°C with g under nitrogen A 1.11 M lithium diisopropylamide/n—hexane— tetrahydrofiiran solution (105 mi, 116 mmol) was added dropvvise over 20 minutes so that the internal temperature did not exceed ~64.3°C. The e was stirred at - 0°C for 1 hour, and tetrahydrofuran (30 ml) and utyl bromoacetate (26.6 g, 136 mmol) were then added dropwise over 10 minutes so that the internal ature did not exceed -60°C. The reaction mixture was stirred at -70°C for further one hour, and a saturated aqueous um chloride solution was then added to the reaction e. Immediately thereafter, flue reaction mixture was diluted with water and ethyl e was added. The organic layer was washed with brine and a 5 N aqueous hydrochloric acid solution, and then FP11-0417 dried over magnesium sulfate, filtered and concentrated. The e was purified by silica gel column chromatography (elution solvent: ethyl acetate/heptane). The residue was fiirther purified by NH silica gel column chromatography (elution solvent: heptane/ethyl acetate = 98/2) to give the title compound (6 g, yield: 14.5%). 1H—NMR (400 MHz, CDC13) 5 ppm; 0.86 (3H, d, J=6 Hz), 1.34 (9H, s), 2.05—2.15 (2H, m), 2.53 (1H, d, J=17 Hz), .00 (3H, m), 3.28 (1H, d, J=10 Hz), 3.59-3.72 (2H, m), 5.08- .16’(2H, m), 7.19—7.39 (10H, m).

By a similar method, 18.1 g ofthe title compound was obtained.

(Example 1h) 1,3—Dibenzyl '(3RS,4SR)[2-(tert-butoxy)oxoethyl]—4— 1‘0 methylpyrrolidine—l,3—dicarboxylate Benzyl (3RS,4SR)benzyl[2-(tert—butoxy)—2—oxoethyl]—4-methylpyrrolidine—3- carboxylate obtained in a manner similar to that in Example 1g (11.7 g, 27.6 mmol) was dissolved in dichloromethane (117 ml), and benzyl chloroformate (23.7 ml, 166 mmol) was added dropwise to the on liquid over 20 minutes so that the al temperature did not exceed 22°C. The mixture was stirred at room temperature for 12 hours, and then solvent was distilled ofi‘. The residue was purified by NH silica gel column chromatography (elution solvent: ethyl acetate/heptane) to give the title compound (9.1 g, yield: 70.5%). 1H—NMR (400 MHZ, CDC13) 5 ppm; 0.84-0.90 (3H, m), 1.32—1.46 (9H, m), 2.09-2.16 (1H, m), 2.21—2.27 (1H, m), 3.04—3.14 (2H, m), 3.33-3.38 (1H, m), 3.60-3.68 (1H, m), 4.32 (1H, t, J=12 Hz), 507-5.20 (4H, m), 7.26-7.36 (10H, m).

Analysis by HPLC; (Analysis conditions 1) Column: PAK AD-H (manufactured by Daicel al Industries, Ltd.) (0.46 cm diameter x 15 cm), eluent: hexane/ethanol = 95/5 (v/v), flow rate: 1 ml/min., detection: UV (210 nm) (Analysis result) The resulting title compound was analyzed under the above analysis conditions 1, and a peak with a retention time of 8.56 s and a peak with a retention time of 1085 minutes were observed.

The title compound separately obtained was analyzed in a chiral column different from the above .

(Analysis ions 2) Column: CHIRALPAK IA (manufactured by Daicel Chemical Industries, Ltd.) (0.46 cm diameter x 15 cm), : hexane/ethanol = 95/5 (V/v), flow rate: 1 ml/min, detection: UV (210 nm) (Analysis result) The resulting title compound was analyzed under the above analysis ions 2, and a peak with a retention time of 6.78 minutes and a peak with a retention time of8.20 minutes were observed.

(Example 1i) )[2-(tert-Butoxy)-2—oxoethyl]methylpyrrolidine—3- carboxylic acid 1,3-Dibenzy1 (3RS,4SR)[2-(tert—butoxy)—2-oxoethyl]—4-methy1pyrrolidine—1,3— dicarboxylate ed in Example lb (9.1 g) was optically resolved repeatedly under the following two types ofconditionsA or B.

Optical resolution by HPLC; (Fractionation conditions A) Column: CHIRALPAK AD—H (manufactured by Daicel Chemical Industries, Ltd.) (2 cm diameter x 25 cm), eluent: hexane/ethanol = 85/15 (V/V), flow rate: 8—10 mI/min.

(Fractionation conditions B) Column: CHIRALPAK IA (manufactmed by Daicel Chemical Industries, Ltd.) (3 cm diameter x 25 cm), eluent: hexane/ethanol = 95/5 (v/v), flow rate: 22 ml/min.

The peak with a shorter retention time was fractionated, and the resulting three lots were then analyzed under the following is conditions.

Analysis by HPLC; sis conditions) Column: CHIRALPAK AD—H (manufactured by Daicel Chemical Industries, Ltd.) (0.46 cm diameter x 15 cm), eluent: hexane/ethanol = 95/5 (v/v), flow rate: 1 ml/min., detection: UV (210 nm) (Analysis result) The retention time was 9.0 minutes to 9.3 s, and the enantiomeric excess was >99% ee for all lots.

The three lots were combined, the resulting chiral form (4.04 g) was dissolved in methanol (121 ml), 10% Pd/C (0.77 g) was added, and the atmosphere was replaced with hydrogen gas. The mixture was stirred at room temperature for 13 hours and then stirred with addition ofwarm water (30 to 40°C, 122 ml), and the precipitated solid was dissolved.- Afier Pd/C was filtered off, the solvent was concentrated and dried to give the title nd (2.1 g).

IR (400 MHz, D20) 5 ppm; 0.97 (3H, d, J=7 Hz), 1.42 (9H, s), 2.15-2.22 (1H, m), 2.30 (1H, d, J=17 Hz), 2.93 (1H, d, J=17 Hz), 3.04 (1H, t, J=12 Hz), 3.18 (1H, d, J=12 Hz), 3.49 (1H, dd, J=8, 12 Hz), 4.03 (1H, d, J=12 Hz).

(Example 1j) (3S,4R)Benzyl[2-(tert-butoxy)oxoethyl]-4— methylpyrrolidine—3-carboxylic acid FP11—0417 A mixture of (3S,4R)-3—[2-(tert-butoxy)—2—oxoethyl]~4—methylpyrrolidine—3- carboxylic acid obtained in e 1i (1.8 g, 7.4 mmol), benzaldehyde (1.51 ml, 14.8 mmol), acetic acid (0.635 ml, 11.1 mmol), sodium toxyborohydride (3.14 g, 14.8 mmol) and methanol (35 ml) was heated at 40°C for 38 hours and 30 s. The on mixture was concentrated, and the resulting residue was purified by silica gel column chromatography (ODS silica gel, elution solvent: water/methanol) to give the title compound as LotA (584 mg) and Lot B’(708 mg). 1H—NMR (400 MHz, CDC13) of Lot A 8 ppm; 1.02 (3H, d, J=7 Hz), 1.38 (9H, s), 2.14 (1H, d, J=17 Hz), 2.15-2.28 (1H, br), 2.97 (1H, d, J=17 Hz), 3.10—3.42 (3H, m), 4.00- 4.10 (1H, m), 4.30—4.40 (1H, br), 4.46 (1H, d, J=12 Hz), 7.45-7.53 (5H, m). lH—NMR ofLot B: identical to NMR ofLotA.

(Example 1k) Chiral form of tert—butyl 2-[(3 S,4R)—1—benzyl—3~{[1—(2~ entyl)piperidin—4—yl]carbamoyl}—4—methylpyrrolidin—3—yl]aeetate A e of (3S,4R)~1~benzyl—3-[2—(tert—butoxy)—2—oxoethyl]—4— methylpyrrolidinecarboxylic acid obtained in Example 1j (300 mg, 0.9 mmol), the chiral form of 1-(2-fluoropentyl)piperidin—4—amine obtained in Example 1d (182 mg, 0.967 mmol), triethylamine (0.376 ml, 2.7 mmol), PyBOP (656 mg, 1.26 mmol) and N,N- dimethylfonnamide (4.5 ml) was d at room temperature for 61 hours and 30 minutes.

Triethylamine (0.207 ml, 1.49 mmol) and PyBOP (360 mg, 0.69 mmol) were further added, followed by stirring for four hours and 30 minutes. Water was added to the reaction mixture, which was extracted with ethyl acetate twice. The organic layer was washed with a saturated aqueous ammonium chloride solution and dried over ous magnesium e, and the solvent was distilled off. The residue was purified by silica gel column chromatography (NH silica gel, elution solvent: ethyl acetate/heptane) to give the title compound (161 mg, yield 35.5%). lH—NIVIR (400 MHz, CDC13) 8 ppm; 0.92 (3H, d, J=7 Hz), 0.96 (3H, t, J=7 Hz), 1.40 (9H, s), 1.40-1.60 (6H, m), 1.82—2.09 (4H, m), 2.16-2.28 (2H, m), 2.36 (1H, d, J=10 Hz), 2.35-2.50 (1H, m), 2.56-2.67 (3H, m), 2.80-2.90 (2H, br), 3.09 (1H, d, J=16 Hz), 3.58379 (4H, m), 4.59-4.80 (1H, m), 7.25—7.40 (5H, m), 8.58 (1H, d, J=8 Hz).

[0158] (Example 11) Chiral form of tert-butyl 2-[(3S,4R)—1-{[2-chloro—6— (uifluoromethyl)phenyl]methyl}{[1-(2-fluoropentyl)piperidin—4-y1]carbamoyl} methylpyrrolidinyl]acetate A mixture of the chiral form of tert-butyl 2—[(3S,4R)-1—benzyl—3-{[1-(2— FP11—0417 fluoropentyl)piperidin—4~yl]carbarnoyl}-4—methylpyrrolidin—3-yl]acetate obtained in Example 1k (161 mg, 0.32 mrnol), 20% palladium hydroxide (135 mg) and methanol (5 ml) was stirred at room ature for 15 hours under a hydrogen atmosphere. The atmosphere in the reaction vessel was replaced with nitrogen, 20% palladium hydroxide was filtered OE, and the solvent was distilled off. 2—(Bromomethyl)—1-chloro-3— (trifluoromethyDbenzene (159 mg, 0.581 , ium carbonate (97 mg, 0.702 mmol) and N,N-dimethylformamide (2 ml) were added to the ing residue, which was heated at 45°C for three hours and 15 minutes. Water was added to the reaction mixture, which was extracted with ethyl acetate twice. The c layer was washed with a saturated aqueous ammonium chloride solution and dried over anhydrous magnesium sulfate, and the solvent was distilled ofi‘. The residue was d by silica gel column chromatography (NH silica gel, elution solvent: ethyl acetate/heptane) to give the title compound (153 mg, yield 78.9%). 1H—NMR (400 MHz, CDC13) 8 ppm; 0.91 (3H, d, J=7 Hz), 0.95 (3H, t, J=7 Hz), 1.38~1.70 (7H, m), 1.39 (9H, s), 1.82-1.90 (1H, br), 1.97 (1H, d, J=16 Hz), 2.00—2.20 (3H, m), 2.31- 2.45 (1H, m), 2.50-2.68 (3H, m), 2.80-2.92 (3H, m), 3.12 (1H, d, J=16 Hz), 3.57 (1H, d, J=1O Hz), 3.60—3.72 (1H, m), 3.92 (1H, d, J=13 Hz), 3.98 (1H, d, J=13 Hz), 4.56-4.74 (1H, m), 7.37 (1H, t, J=8 Hz), 7.61-7.68 (2H, m), 7.85 (1H, d, J=8 Hz).

(Example 1m) Chiral form of 2—[(3S,4R)—l—{[2-chloro (trifluoromethyl)phenyl]methyl}{[l-(2-fluoropentyl)piperidin—4—yl]carbamoyl}—4— methylpyrrolidin—3-y1]acetic acid A mixture of the chiral form of tert—butyl 2-[(3S,4R)—1-{[2—chloro-6— oromethyl)phenyl]methyl}-3—{[1-(2-fluoropentyl)piperidin—4—yl]carbamoyl}-4— methylpyrrolidin—3-yl]acetate obtained in Example 11 (153 mg, 0.252 mmol), trifluoroacetic acid (2 ml, 26.9 mmol) and dichloromethane (dehydrated) (2 ml) was stirred at room temperature for two hours and 35 minutes. The reaction mixture was concentrated, and the ing residue was purified by silica gel column tography (ODS silica gel, elution t: water/methanol) to give the title compound (122 mg, yield 88%). 1H—NMR (400 MHz, CDC13) 8 ppm; 0.94—1.02 (6H, m), 1.32—1.70 (5H, m), 1.82-1.90 (1H, br), 2.07-2.20 (3H, m), 2.35-2.75 (8H, m), 2.81-3.05 (3H, m), 3.19 (1H, d, J=10 Hz), 3.63- 3.75 (1H, m), 3.89 (1H, d, J=13 Hz), 4.00 (1H, d, J=13 Hz), 4.56-4.76 (1H, m), 7.40 (1H, t, J=8 Hz), 7.62-7.70 (2H, m), 8.52 (1H, d, J=7 Hz).

(Example 2) 2—[(3S,4R)[(2,6-Dichloropheny1)methyl]({l-[(4,4- difluorocyclohexyl)methyl]piperidin—4-yl}carbarnoyl)—4-methylpyrrolidin—3-yl]acetic acid (Example 2a) 4,4-Difluoro—N—methoxy—N~methylcyclohexane—l-carboxamide A mixture of ethyl 4,4-difluorocyclohexane-l-carboxylate (1.9 g, 9.88 mmol) and tetrahydrofiiran (60 ml) was cooled to -70°C, and N,O-dimethylhydroxyamine hydrochloride (1.44 g, 14.8 mmol) was added. r, 1.05 M n—propylmagnesium bromide (24.2 ml, 25.2 mmol) was added dropwise to the reaction mixture at -55°C over five minutes so that the internal temperature did not exceed -35°C, and the reaction mixture was stirred at 0°C for 30 s. A saturated aqueous ammonium chloride solution was added to the reaction mixture, which was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate, and the solvent was distilled otf. The residue was purified by silica gel column chromatography (elution solvent: ethyl e/heptane) to give the title compound (1.7 g, yield 83%). 1H—NMR (400 MHz, CDC13) 5 ppm; 1.65-1.95 (6H, br), 2.13—2.30 (2H, br), 2.70~2.82 (1H, br), 3.20 (3H, s), 3.70 (3H, s).

(Example 2b) 4,4-Difluorocyclohexane—1~carba1dehyde A mixture of 4,4—difluoro-N-methoxy—N—methylcyclohexane—1-carboxamide obtained in e 2a (1.7 g, 8.21 mmol) and tetrahydrofinan (60 ml) was cooled to ~70°C, a 1.0 M diisobutylaluminum hydride/toluene solution (9.85 ml, 9.85 mmol) was added, and the reaction mixture was stirred at -60°C for 35 s. A 1.0 M diisobutylaluminum hydride/toluene solution (5 ml, 5 mmol) was finther added at -65°C, and the reaction mixture was stirred at —70°C for two hours. A2 N aqueous hydrochloric acid solution was added to the reaction mixture, which was ted with ethyl acetate. The organic layer was dried over magnesium sulfate, and the solvent was distilled off to give a crude t of the title compound (1.4 g). 1H—NMR (400 MHz, CDC13) 5 ppm; 1.72-1.92 (4H, br), 1.95—2.18 (4H, br), 2.30-2.40 (1H, br), 9.68 (1H, s).

(Example 20) tert-Butyl N—{1-[(4,4-difluorocyclohexyl)methyl]piperidin—4- yl}carbamate A mixture of the crude product of 4,4—difluorocyclohexane—1~carbaldehyde obtained in Example 2b (1.4 g) and tetrahydrofiiran (100 ml) was cooled to 0°C, tert-butyl N- (piperidin—4—yl)carbamate (2.27 g, 11.3mmol) was added thereto, followed by stirring for 20 minutes. Sodium triacetoxyborohydride (2.2 g, 10.4 mmol) was then added to the on mixture, which was stirred at room temperature for eight hours. Brine was added to the reaction mixture, which was extracted with ethyl acetate. The organic layer was dried over FP11~0417 magnesium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography (elution t: ethyl acetate/heptane). The resulting residue was dissolved in ethyl acetate and washed with a saturated aqueous sodium bicarbonate solution, and the solvent was then distilled ofito give the title compound (650 mg, yield 20.7%). 1H—NMR (400 MHz, CDC13) 5 ppm; 1.15-2.20 (17H, m), 1.44 (9H, s), 2.70-2.85 (2H, br), 3.38-3.53 (1H, br), .50 (1H, br).

(Example 2d) l-[(4,4-Difluorocyclohexyl)methyl]piperidin—4-amine A 4 N hydrogen de/1,4—dioxane solution (11 ml, 43 mmol) was added to a e of tert—butyl N~{l~[(4,4—difluorocyclohexyl)methyl]piperidin—4—yl}carbarnate ed in Example 2c (650 mg, 1.96 mmol) and methanol (11 ml), followed by stirring at room temperature for three hours. After concentrating the reaction liquid, the residue was dissolved in ethyl acetate, a l N aqueous sodium hydroxide solution was added, and the organic layer was separated. The aqueous layer was flirther extracted with ethyl acetate twice, the organic layers were dried over sodium sulfate, and the solvent was then distilled 01f to give the title compound (419 mg, yield 92%). 1H—NMR (400 MHZ, CDC13) 8 ppm; 1.16-1.42 (7H, m), 1.59-2.16 (12H, m), .68 (1H, m), 2.74-2.82 (2H, m).

(Example 2e) (4-Methoxypheny1)methyl(2E)—but—2-enoate Crotonic acid (17.2 g, 200 mmol) was ved in NN—dimethylfonnamide (100 ml) with cooling in an ice bath under nitrogen, and powdered potassium carbonate (15.2 g, 110 mmol) was added. The mixture was d for 30 minutes, and 4—methoxybenzyl chloride (29.8 g, 190 mmol) was then added dropwise over 15 minutes. The reaction mixture was stirred at room temperature for four hours and at 45°C for 14 hours. Ethyl acetate (500 ml) and water (200 ml) were added to the reaction liquid. The separated organic layer was washed with water (100 ml x 2) and brine (100 ml). The aqueous layer was extracted with ethyl acetate (100 ml). The organic layers were combined, dried over magnesium sulfate, filtered and concentrated under reduced re to give a crude product ofthe title compound (40.51 g, yield: 98.2%). 1H—NMR (400 MHz, CDC13) 8 ppm; 1.87 (3H, dd, J=2, 7 Hz), 3.81 (3H, s), 5.10 (2H> s), 5.87 (1H, dq, J=2, 16 Hz), 6.86-6.92 (2H, m), 7.00 (1H, dq, J=7, 16 Hz), .34 (2H, m).

(Example 2f) (4-Methoxyphenyl)methyl (3RS,4SR)benzyl-4— methylpyrrolidine—3—carboxylate The title nd (57.86 g, yield: 86.5%) was obtained from (4- FPll-0417 methoxyphenyl)methyl ut-2~enoate obtained in Example 2e (40.6 g, 197 mmol) by a method similar to the method described in Example 1f. 1H-NMR (400 MHz, CDC13) 6 ppm; 1.12 (3H, d, J=7 Hz), 2.17—2.24 (1H, m), 2.46-2.62 (2H, m), 2.73-2.86 (2H, m), 2.87—2.93 (1H, m), 3.55 (1H, d, J=13 Hz), 3.63 (1H, d, J=l3 Hz), 3.81 (3H, s), .10 (2H, m), 6.86—6.92 (2H, m), .35 (7H, m).

(Example 2g) (4—Methoxyphenyl)methyl (3RS,4SR)-l~benzyl-3—[2—(tert—butoxy)- 2-ox0ethyl]methy1py1rolidine—3-carboxy1ate The title compound (60.3 g, yield: 80.1%) was obtained fiom (4- methoxyphenyl)methyl SR)-1—benzyl—4—methylpyrrolidine~3-carboxylate obtained in Example 2f(56.7 g, 166 mmol) by a method similartothe method described ’i‘n'Example 1g. 1H—NMR (400 MHz, CDC13) 8 ppm; 0.83 (3H, d, J=7 Hz), 1.35 (9H, s), 2.05—2.14 (2H, m), 2.51 (1H, d, J=17 Hz), 2.90-2.98 (2H, m), 3.60 (1H, d, .1213 Hz), 3.70 (1H, d, J=13 Hz), 3.81 (3H, s), 5.04 (1H, d, J=12 Hz), 5.07 (1H, d, J=12 Hz), 6.85-6.91 (2H, m), 7.19-7.34 (7H, m).

(Example 2h) l-Benzyl 3—(4—methoxyphenyl)methyl (3RS,4SR)-3—[2~(te1t— butoxy)-2—oxoethy1]methylpyrrolidine—1,3-dicarboxylate The title compound (15 g, yield: 65.1%) was obtained from (4- methoxyphenyl)methyl (3RS,4SR)benzyl[2-(tert—butoxy)oxoethyl]~4- methylpyrrolidinecarboxylate obtained in Example 2g (21 g, 46.3 mmol) by a method similar to the method described in e 1h. 1H—NMR (400 MHz, CDC13) 5 ppm; .88 (3H, m), 1.38 (9H, d, J=5 Hz), 2.04-2.15 (1H, m), 2.22 (1H, dd, J=3, 7 Hz), .12 (2H, m), 3.30-3.36 (1H, m), 3.57-3.67 (1H, m), 3.79 (3H, d, J=4 Hz), 4.30 (1H, t, J=12 Hz), 5.00-5.16 (4H, m), 6.82—6.86 (2H, m), 7.23—7.39 (7H, m).

Analysis by HPLC; (Analysis conditions) Column: CHIRALPAK AD—H (manufactured by Daicel Chemical Industries, Ltd.) (0.46 cm diameter x 15 cm), 40°C, eluent: hexane/ethanol = 9/1 (V/V), flow rate: 1 ml/min., detection: UV (210 nm) (Analysis result) The resulting title compound was analyzed under the above is conditions, and a peak with a retention time of7.14 minutes and a peak with a retention time of9.06 minutes were observed.

(Example 2i) l-Benzyl 3-(4-methoxyphenyl)methyl (3S,4R)-3~[2~(tert—butoxy)-2— oxoethyl]-4~methylpyrrolidjne-l,3-dicarboxylate l-Benzyl 3—(4-methoxyphenyl)methyl (3RS,4SR)[2-(te1t—butoxy)—2—oxoethyl]— FP11-0417 4—methylpyrrolidine—l,3-dicarboxylate ed in Example 2h (7.2 g, 14.5 mmol) was optically resolved repeatedly by HPLC (CHIRALPAK AD-H (2 cm er x 25 cm), elution solvent: hexane/ethanol = 85/15, flow rate: 8-10 ml/min) to give a chiral form corresponding to the peak with a shorter retention time (2.92 g, yield: 40.5%).

Analysis by HPLC; sis conditions) Column: CHlRALPAK AD-H (manufactured by Daicel Chemical Industries, Ltd.) (0.46 cm er x 15 cm), 40°C, eluent: hexane/ethanol = 9/1 (V/v), flow rate: 1 ml/min., ion: UV (210 nm) (Analysis result) The resulting chiral form was analyzed under the above analysis conditions to find”thatthe retention time was 7.18 minutes e enantiome'ric excess was >99% ee.

(Example 2j) (3S,4R)—3-[2-(tert—Butoxy)—2-oxoethyl]methylpyrrolidine—3— carboxylic acid Ethanol (50 ml) and 10% Pd/C (500 mg) were added to l-benzyl 3-(4- methoxyphenyl)methyl (3S,4R)—3~[2—(tert—butoxy)-2—oxoethyl]—4-methylpyrrolidine—1 ,3- .15 ’ dicarboxylate ed in Example 2i (2.92 g, 5.87 mmol), followed by stirring at room temperature for 38 hours and five minutes under a hydrogen atmosphere. Water (100 ml) was added to the reaction liquid, which was stirred and filtered. The filtrate was concentrated to give the title compound (1.37 g, yield: 95.9%). 1H—NMR (400 MHZ, D20) 5 ppm; 1.00 (3H, d, J=7 Hz), 1.45 (9H, s), 2.16-2.37 (1H, m), 2.33 (1H, d, J=17 Hz), 2.96 (1H, d, J=17 Hz), 3.05—3.11 (1H, m), 3.22 (1H, d, J=12 Hz), 3.50—3.56 (1H, m), 4.07 (1H, d, J=12 Hz).

(Example 2k) (3S,4R)[2-(tert—Butoxy)—2—oxoethyl]-1—[(2,6- dichlorophenyl)methyl]-4—methylpyrrolidine—3—carboxylic acid 2,6—Dichlorobenzaldehyde (646 mg, 3.7 mmol) and sodium triacetoxyborohydn'de (782 mg, 3.7 mmol) were added to a mixture of (3S,4R)—3-[2-(tert—butoxy)—2—oxoethyl]~4- methylpyrrolidine—3-carboxylic acid obtained by the method of Example 2j (600 mg, 2.47 mmol), acetic acid (0.14 ml, 2.47 mrnol) and methanol (10 ml), and the reaction mixture was stirred at room ature for two hours. 2,6—Dichlorobenzaldehyde (430 mg, 2.5 mmol) and sodium triacetoxyborohydride (522 mg, 2.5 mrnol) were further added, followed by stirring at an al temperature of40°C for 10.5 hours. Water was added to the reaction mixture, which was extracted with dichloromethane five times, and the organic layer was concentrated. The residue was purified by silica gel column chromatography (elution solvent: ethyl acetate/methanol) to give Lot A (32 mg) and Lot B (551 mg) of the title FP11—0417 compound.

The chemical shift of Lot A is shown below. Lot B is a combination of thee fiactions upon column purification, and they were combined after confirming that the chemical shift ofeach fiaction is similar to the chemical shift ofLot A. 1H—NMR (400 MHz, CDCl}) 6 ppm; 1.01 (3H, d, J=7 Hz), 1.42 (9H, s), 2.05—2.11 (1H, m), 2.16—2.22 (ll-I, m), .69 (2H, m), 2.97—3.04 (2H, m), 3.70 (1H, d, J=10 Hz), 4.07—4.15 (2H, m), 7.22 (1H, dd, J=8, 9 Hz), 7.35 (2H, d, J=8 Hz).

(Example 21) tert—Butyl 2—[(3S,4R)—1-[(2,6-dichlorophenyl)methyl]—3-({1-[(4,4— difluorocyclohexyl)methyl]piperidin—4—yl}carbamoyl)—4—mefl1ylpyrrolidin—3-yl]acetate The title compound (409 mg, yield: 92%) was obtained from l~[(4,4— difluorocyclohexyl)methyl]piperidin—4~amine ed in Example 2d (201 mg, 0.87 mmol) and (3S,4R)~3-[2—(tert~butoxy)—2—oxoethyl]-1—[(2,6-dichlorophenyl)methyl]-4— methylpyrrolidine—3-carboxylic acid obtained in Example 2k (290 mg, 0.72 mmol) by a method similar to the method described in Example 1k 1H—NMR (400 MHz, CDC13) 5 ppm; 0.85-0.93 (3H, m), 1.15-2.14 (19H, m), 1.41 (9H, s), 2.57-2.71 (4H, m), 2.92 (1H, t, J=10 Hz), 3.12 (1H, d, J=16 Hz), 3.59-3.72 (2H, m), 3.96 (2H, s), 7.18 (1H, t, J=8 Hz), 7.33 (2H, d, J=8 Hz), 8.11 (1H, d, J=8 Hz).

(Example 2m) 2—[(3S,4R)-1—[(2,6-Dichlorophenyl)methyl]—3—({1-[(4,4- difluorocyclohexyl)methy1]pipen'din—4—yl}carbamoyl)-4—methylpy1rolidin—3—yl]acetic acid The title compound (242 mg, yield: 66%) was obtained from text—butyl ,4R)- l-[(2,6-dichlorophenyl)methy1]—3-({ l-[(4,4-difluorocyclohexyl)methyl]pipen'din—4- bamoyl)—4—methylpyrrolidin—3-yl]acetate obtained in Example 21 (409 mg, 0.66 mmol) by a method similar to the method bed in Example 1m. 1H—NMR (400 MHz, CD3OD) 5 ppm; 0.87-0.93 (3H, m), 1.20-2.38 (19H, m), 2.62-2.71 (2H, m), 2.86—3.12 (4H, m), 3.61-3.73 (2H, m), 4.01—4.09 (2H, m), 7.29 (1H, dd, J=8, 9 Hz), 7.43 (2H, d, J=8 Hz).

(Example 3) 2-[(3 S,4R)—1~{[2—Chloro-6—(tiifluoromethyl)phenyl]methyl}-3—{[1— (cyclohex—l -en-1 —ylmethyl)piperidin—4—yl]carbamoyl}-4—methylpy1rolidin—3—yl]acetic acid le 3a) utyl 4-[(3S,4R)benzyl[2-(tert—butoxy)-2—oxoethy1] methylpyrrolidin—3-amido]piperidine—l-carboxylate text-Butyl 4-aminopiperidine—1-carboxylate (849 mg, 4.24 mmol), triethylamine (1.18 ml, 8.48 mmol) and PyBOP (2.21 g, 4.24 mmol) were added to a solution of (3S,4R)- 1~benzyl—3-[2-(tert—butoxy)—2~oxoethyl]—4-methylpyrrolidine—3-carboxylic acid obtained by a method similar to that of Example 1j (942 mg, 2.83 mmol) in N,N-dimethy1formamide (20 ml), followed by stirring at room temperature overnight. Ethyl acetate was added to the reaction liquid, which was washed with a 1 N aqueous sodium hydroxide solution and brine.

This was dried over anhydrous magnesium sulfate and concentrated. The residue was purified by silica gel column chromatography on solvent: heptane/ethyl acetate) to give the title compound (1.33 g, yield: 91.1%). \4R’(400‘Ilez,"CDC13) 5 ppm; 0.92’(3H, d, J=7 Hz), 1.20-1.40‘(2H, m), 1.40 (9H, s), 1.48 (9H, s), .88 (1H, m), 1.92-1.98 (1H, br), 1.96 (1H, d, J=16 Hz), 2.03—2.09 (1H, m), 2.36 (1H, d, J=10 Hz), 2.58-2.68 (2H, m), 2.89—2.95 (2H, m), 3.10 (1H, d, J=16 Hz), 3.59 (1H, d, J=10 Hz), 3.66 (2H, s), .00 (3H, m), 7.23-7.35 (5H, m), 8.65 ‘(1H, d, J=7 Hz).

MS (ESI) m/z: 538.2 (M+Na)+.

(Example 3b) tert-Butyl 4—[(3S,4R)—3-[2—(tert—butoxy)—2~oxoethyl]—4~ methylpyrrolidin—3-amido]piperidine—1carboxylate % palladium hydroxide (724 mg) was added to a solution of tert-butyl 4— [(38,4R)-l~benzyl—3—[2-(tert—butoxy)—2-oxoethyl]methylpyrrolidin—3-amido]piperidine~l- carboxylate obtained in Example 3a (1.33 g, 2.58 mmol) in methanol (30 ml), which was stirred under a hydrogen atmosphere overnight The reaction liquid was filtered and concentrated under reduced pressure to give the title compound (1.04 g, yield: 94.7%). 1H—NMR (400 MHZ, CDC13) 5 ppm; 0.92 (3H, d, J=7 Hz), 1.20-1.52 (2H, m), 1.41 (9H, s), 1.43 (9H, s), 1.80—2.10 (5H, m), 2.00 (1H, d, J=16 Hz), 2.55-2.61 (1H, m), 2.80—3.06 (2H, m), 2.92 (1H, d, J=10 Hz), 3.12 (1H, d, J=16 Hz), 3.35 (1H, d, J=9 Hz), 3.70 (1H, d, J=10 Hz), 3.80—4.00 (3H, m), 8.30 (1H, d, l=7 Hz).

MS (ESI) m/z: 426.1 (M+H)+. le 3c) tert-Butyl 4—[(3 S,4R)—3—[2-(tert~butoxy)-2—oxoethyl]~1—{[2-chloro (tnfluoromethyl)pheny1]methyl}—4-methylpyrrolidin—3—amido]piperidine-l-carboxylate 2-(Bromomethyl)—1-chloro(t1ifluoromethyl)benzene (443 mg, 1.62 mmol) and potassium carbonate (244 mg) were added to a solution of tert—butyl 4-[(3S,4R)[2-(tert— )-2—oxoethyl]—4-methylpyrrolidin—3—amido]piperidine—1—carboxylate obtained in Example 3b (345 mg, 0.811 mmol) in N,N—dimethylformamide (dehydrated) (10 mL), 3O which was stirred at 45°C for six hours and at 40°C for two days. Ethyl acetate was added to the reaction liquid, which was washed with a l N s sodium hydroxide solution and brine. This was dried over anhydrous magnesium sulfate and trated. The residue was purified by silica gel column tography (elution solvent: heptane/ethyl acetate) to FP11-0417 give the title nd (320 mg, yield: 63.8%). 1H-NMR (400 MHz, CDC13) 5 ppm; 0.92 (3H, d, J=7 Hz), 1.13-1.18 (2H, m), 1.39 (9H, s), 1.49 (9H, s), 1.53—1.65 (1H, m), 1.76-1.86 (1H, m), 1.98 (1H, d, J=16 Hz), 2.02-2.10 (1H, m), 2.51 (1H, d, J=10 Hz), 2.60-2.80 (3H, m), 2.88 (1H, t, J=10 Hz), 3.12 (1H, d, J=16 Hz), 3.53 (1H, d, J=10 Hz), 4.09-4.25 (5H, m), 7.38 (1H, t, J=8 Hz), 7.63 (2H, d, J=8 Hz), 7.96 (1H, d, J=8 Hz).

MS (1381) m/z: 640.2 (M+Na)+.

(Example 3d) 2—[(3S,4R)—1-{[2-Chloro—6-(trifluoromethyl)phenyl]methyl}~4- methyl—3-[(piperidin—4—yl)carbamoyl]pyrrolidin—3—yl]acetic acid Trifluoroacetic acid (8 mL) was added to a solution of tert—butyl 4-‘[(3S,4R)[2— (tert-butoxy)-2—oxoethyl]—1~{[2—chloro—6—(trifluoromethyl)phenyl]methyl}—4- pyrrolidin—3-amido]piperidine—l-carboxylate obtained in Example 3c (320 mg, 0.518 mmol) in dichloromethane (dehydrated) (8 mL) under ice-cooling, followed by stirring at room temperature for 2.5 hours. The on liquid was concentrated under reduced ‘15 pressure, and the residue was purified by ODS column chromatography (elution solvent: water/methanol) to give a mixture containing the title nd (344 mg). 1H—NMR (400 MHz, CD30D) 6 ppm; 0.83 (3H, d, J=5 Hz), .63 (2H, m), 1.76-1.89 (1H, m), 1.92-2.00 (1H, m), 2.03-2.19 (2H, m), 2.55—2.68 (2H, m), 2.91-3.10 (4H, m), 3.25— 3.36 (2H, m), 3.45-3.59 (1H, m), 3.75—4.18 (3H, m), 7.41—7.76 (3H, 111).

MS (ESI) m/z: 462.3 (M+H)+ (Example 3e) 2—[(3S,4R)~1~{[2—Chloro(trifluoromethyl)phenyl]methyl}—3-{[1- (cyclohex—l—en—l-ylmethyl)piperidin—4—yl]carbamoyl}—4—methylpyrrolidin—3-yl]acetic acid ex—l~ene-1—carbaldehyde (423 1.11, 3.73 mmol), acetic acid (300 pl) and sodium triacetoxyborohydride (789 mg, 3.73 mmol) were added to a solution ofthe mixture of 2—[(3S,4R)—1~{[2—chloro-6—(trifluoromethyl)phenyl]methyl}~4—methy1—3-[(piperidin—4— yl)carbamoyl]pyrrolidin—3-yl]acetic acid obtained by the method of Example 3d (344 mg, 0.745 mmol) in tetrahydrofiiran (dehydrated) (10 mL), followed by stirring overnight.

Water and methanol were added to the reaction liquid, which was trated under reduced re, and the residue was purified by ODS column chromatography (elution solvent: water/methanol). The purified product was dissolved in dichloromethane, suspended by adding hexane and concentrated to give the title compound (180 mg, yield: 43.4%). 1H—NMR (400 MHZ, CD3OD) 8 ppm; 0.89 (3H, d, J=7 Hz), 1.23—1.38 (2H, m), 1.44-1.82 FP11-0417 (6H, m), .96 (1H, m), 1.96-2.25 (5H, m), 2.30-2.45 (2H, m), 2.55-2.68 (2H, m), 2.92- 3.20 (5H, m), 3.54 (1H, d, J=10 Hz), 3.64—3.78 (1H, m), 3.95 (1H, d, J=10 Hz), 4.05 (1H, d, J=10 Hz), 5.76 (1H, s), 7.47-7.52 (1H, m), 7.72 (1H, d, J=7 Hz), 7.77 (1H, d, J=8 Hz).

MS (ESI) m/z: 578.3 (M+Na)+ (Example 4: Another method for compound of Example 3) 2~[(3S,4R)—1-{[2— Chloro(trifluoromethyl)phenyl]methyl}-3—{[1-(cyclohex—l-en-1—ylmethyl)piperidin—4— yl]carbamoyl}-4—methylpyrrolidin—3-yl]acetic acid (Example 4a) tert—Butyl N—[1-(cyclohex—l—en—1~ylmethyl)piperidin—4—yl]carbamate A mixture of tert-butyl eridin—4—yl)carbamate (5.3 g, 26.5 mmol), l— cyclohexene—l-carboxaldehyde (3.5 g, 31.8 mn’ml), sodium triac’etoxyborohydiide (7.29 g, 34.4 mmol), acetic acid (0.5 ml) and tetrahydrofuran (dehydrated) (80 ml) was stirred at room ature for 85 hours and 30 minutes. Water and sodium bicarbonate were added to the reaction mixture, which was stirred and then extracted with ethyl e twice. The organic layer was washed with brine and dried over anhydrous sodium sulfate, and the solvent was distilled off. The residue was purified by silica gel column chromatography (NH silica gel, elution solvent: ethyl acetate/heptane) to give the title compound (7.47 g, yield: 95.7%). 1H—NMR (400 IVHiz, CDC13) 8 ppm; 1.35—1.50 (2H, m), 1.44 (9H, s), 1.51-1.65 (4H, m), 1.84-2.03 (8H, m), 2.68-2.80 (4H, m), 3.39—3.50 (1H, m), .48 (1H, m), 5.54 (1H, s).

[0182] (Example 4b) l—(Cyclohex—1-en—1—ylmethyl)piperidin—4-amine A e of tert—butyl N—[l—(cyclohexen—1-ylmethy1)piperidin—4~yl]carbamate obtained in Example 4a (7.47 g, 25.4 mmol), a 5 N aqueous hydrochloric acid solution (25 ml, 125 mmol) and methanol (100 ml) was stirred at 70°C for 1.5 hours. The reaction mixture was ice-cooled, a 5 N aqueous sodium hydroxide solution (25 ml, 125 mmol) was added, and the solvent was led off. Water was added to the residue, which was extracted with ethyl acetate twice. The organic layer was washed with brine and dried over anhydrous sodium sulfate, and the t was distilled offto give the title compound (4.73 g, yield: 95.8%). 1H—NMR (400 MHz, CDC13) 6 ppm; 1.30-1.50 (4H, m), 1.51—1.70 (4H, m), 1.75-2.08 (8H, m), 2.59-2.68 (1H, m), 2.72-2.88 (4H, m), 5.55 (1H, s).

(Example 40) l-Benzyl 3-(4—methoxyphenyl)methyl (3RS,4SR)[2-(tert— )oxoethyl]methy1pyrrolidine—1,3—dicarboxylate A solution of (4—methoxyphenyl)methyl (3RS,4SR)—1~benzyl—3—[2—(tert—butoxy)—2— FP11-0417 oxoethyl]—4—methylpyrrolidine—3—carboxylate obtained in Example 2g (18 g, 39.7 mmol) in dichloromethane (85.7 ml) was ed to an internal temperature of 10 to 20°C, and benzyl chloroformate (11.9 ml, 79.4 mmol) was added thereto. After returning to room temperature and ng for 0.5 hour, the on mixture was concentrated directly. The resulting crude product was purified by column chromatography (silica gel, elution solvent: ethyl acetate/heptane) to give the title compound (12.89 g, yield: 65.3%). 1H—NMR (400 MHz, CDC13) 5 ppm; 0.80-0.88 "(3H, m), 1.38 (9H, d, J=5 Hz), 2.04—2.15 (1H, m), 2.22 (1H, dd, J=3, 17 Hz), 3.00—3.12 (2H, m), 3.62 (1H, m), 3.79 (3H, d, J=4 Hz), 4.30 (1H, t, J=12 Hz), 5.00-5.16 (4H, m), 6.82-6.86 (2H, m), 723—739 (8H, m).

[0184] (Example 4d) l—Benzyl ethoxyphenyl)methyl (3S,4R)[2-(tert—butoxy)—2— oxoethyl]—4-methylpyrrolidine—1,3-dicarboxylate l-Benzyl 3—(4-methoxyphenyl)methyl (3RS,4SR)—3—[2-(tert—butoxy)—2~oxoethyl]- 4—methylpyrrolidine—l,3-dicarboxylate obtained in Example 4c (12.7 g, 26 mrnol) was optically ed repeatedly by HPLC (CHIRALPAK AD-H (2 cm diameter x 25 cm), elution solvent: hexane/ethanol = 85/15, flow rate: 8—10 mJ/min.) to give a chiral form corresponding to the peak with a shorter retention time (5.7 g, yield: 44.9%). The resulting chiral form (5.3 g) was fiirther purified by silica gel column chromatography (elution solvent; heptane/ethyl acetate) in two portions to give 5.2 g ofthe title compound. 1H—NMR (400 MHz, CDC13) 8 ppm; 0.81-0.88 (3H, m), 3.37-3.38 (9H, m), 2.04—2.16 (1H, m), 2.19-2.25 (1H, m), .12 (2H, m), 3.30-3.36 (1H, m), 3.56-3.67 (1H, m), 3.75-3.79 (3H, m), 4.26-4.33 (1H, m), 5.00-5.16 (4H, m), 6.82-6.86 (2H, m), 727-7.37 (7H, m).

Analysis by HPLC; sis conditions) Column: CHIRALPAK AD—H (manufactured by Daicel Chemical Industries, Ltd.) (0.46 cm diameter x 15 cm), 40°C, eluent: hexane/ethanol = 9/1 (v/v), flow rate: 1 m]/min., ion: UV (210 nm) (Analysis result) The d chiral form was analyzed under the above analysis conditions to find that the retention time was 7.15 minutes and the enantiomeric excess was >99% ee.

(Example 4e) l-Benzyl 3-(4—methoxyphenyl)methyl (3S,4R)[2~(tert—butoxy)—2— oxoethyl]—4-methylpyrrolidine-l,3-dicarboxylate l-Benzyl 3-(4—methoxyphenyl)methyl (3RS,4SR)[2—(tert—butoxy)—2~oxoethyl]- 4-methylpy1rolidine—l,3—dicarboxylate obtained by a method r to that of e 4c (7.8 g, 16.5 mmol) was optically resolved repeatedly by HPLC (CI-HRALPAK AD-H, elution solvent: hexane/ethanol = 85/15) to give a chiral form with a r retention time FPll-0417 (3.3 g, yield: 42.3%).

(Example 41) (3S,4R)[2-(tert—Butoxy)oxoethyl]-4—methylpy1rolidine ylic acid Methanol (70 ml) and 10% Pd/C (600 mg) were added to l—benzyl 3-(4- methoxyphenyl)methyl (3 S,4R)—3—[2—(tert—butoxy)—2-oxoethyl]—4—methylpyrrolidine—1,3- dicarboxylate obtained by the method ofExample 4e (3.3 g, 6.63 mmol), followed by stirring at room temperature for three days under a hydrogen here. Water (70 ml) was added to the reaction liquid, which was stirred and filtered. The filtrate was concentrated to give the title compound (1.15 g, yield: 71.3%). 1H-NMR (400 MHz, D20) was identical to that ofthe compound ed in Exarnple 2].

(Example 4g) (3S,4R)—3~[2-(tert—Butoxy)oxoethy1]-4—methylpyrrolidine—3- ylic acid Methanol (3 ml) and 10% Pd/C (30 mg) were added to l-benzyl 3-(4- methoxyphenyl)methyl (3S,4R)[2-(tert—butoxy)—2—oxoethyl]—4—methylpyrrolidine—1,3- dicarboxylate which is obtained in Example 4d and which is without silica gel column tography purification (104 mg, 0.21 mmol), followed by stirring at room temperature overnight under a hydrogen atmosphere. Water was added to the reaction liquid, which was stirred and filtered. The filtrate was concentrated to give the title compound (48.7 mg, yield: 95.3%). 1H—NMR (400 MHZ, D20) was cal to that ofthe compound obtained in Example 2j.

(Example 4h) (3S,4R}3-[2—(tert—Butoxy)—2~oxoethyl]—4—methylpyirolidine—3— carboxylic acid Methanol (100 ml) and 10% Pd/C (559 mg) were added to l-benzyl 3-(4- methoxyphenyl)methyl (3S,4R)-3—[2-(tert—butoxy)—2—oxoethyl]—4—methylpyrrolidine—1 ,3~ dicarboxylate ed in Example 4d (5.2 g, 10.5 mmol), followed by stirring at room temperature for eight hours and 15 minutes under a hydrogen atmosphere. Water (100 ml) was added to the reaction liquid, which was stirred for one hour and filtered. The filtrate was concentrated. A combination (3.2 g, 13.2 mmol) ofthe residue and (3S,4R)—3-[2-(tert— )-2—oxoethyl]methylpyrrolidine—3-carboxylic acid obtained in es 2j, 4f and 4g was suspended by adding methanol (25 ml) and stirred at room temperature for 30 minutes, and collection by filtration gave 3 .02 g ofthe title compound. 1H-NMR (400 MHz, D20) 5 ppm; 0.98 (3H, d, J=7 Hz), 1.43 (9H, s), 2.14—2.24 (1H, m), 2.31 (1H, d, J=17 Hz), 2.94 (1H, d, J=17 Hz), 3.06 (1H, t, J=11 Hz), 3.20 (1H, d, J=12 Hz), FP11—0417 3.51 (1H, dd, J=8, 12 Hz), 4.05 (1H, d, J=12 Hz).

(Example 4i) tert—Butyl 2-[(3 S,4R)-l-{[2-chloro (trifluoromethyl)phenyl]methyl}{[1-(cyclohexenylmethy1)piperidin—4- yl]carbamoyl}—4-methy1pyrrolidin—3—yl]acetate lohex~l—enylmefliyl)piperidin—4-amine obtained in Example 4b (240 mg, 1.24 mmol), N,N—dimethylformamide (3 ml), triethylamine (344 pl, 2.47 mmol) and PyBOP (856 mg, 1.64 mmol) were added to (3S,4R)—3—[2-(tert—butoxy)~2~oxoethyl]—4— methylpyrrolidine—3—carboxylic acid obtained in Example 4h (200 mg, 0.822 mmol), followed by stirring with heating in a 45°C oil bath for 1.5 hours. Water was added to the reaction mixture, which was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and concentrated. 2—(Bromomethyl)—1~chloro (trifluoromethyl)benzene (247 mg, 0.903 mmol), potassium carbonate (170 mg, 1.23 mmol) and N,N—dimethy1formarnide (1 ml) were added thereto, followed by heating with stirring in a 45°C oil bath for three hours. Water was added to the reaction mixture, which was extracted With ethyl acetate. The organic layer was washed with water and brine and then dried over magnesium sulfate. The ing crude product was purified by column chromatography twice (NH silica gel, n t: ethyl acetate/heptane and silica gel, elution t: ethyl acetate/heptane) to give the title compound (353 mg, yield 70.1%). 1H—NMR (400 MHz, CDC13) 6 ppm; 0.91 (3H, d, J=7 Hz), 1.31-1.42 (2H, m), 1.39 (9H, s), .66 (5H, m), .09 (9H, m), 2.53 (1H, d, J=10 Hz), 2.63-2.77 (5H, m), 2.88 (1H, t, J=10 Hz), 3.11 (1H, d, J=16 Hz), 3.56 (1H, d, J=10 Hz), 3.61-3.72 (1H, m), 3.92 (1H, d, J=13 Hz), 3.98 (1H, d, J=13 Hz), 5.54 (1H, s), 7.36 (1H, t, J=8 Hz), 7.62 (1H, d, J=4 Hz), 7.64 (1H, d, J=4 Hz), 7.83 (1H, d, J=8 Hz).

(Example 4j) 2—[(3 S,4R)—1-{[2—Chloro—6—(trifluoromethyl)phenyl]methyl}{[1~ (cyclohex—l-en-1~ylmethy1)piperidin—4—yl]carbamoyl}~4~methylpyrrolidin—3-yl]acetic acid Trifluoroacetic acid (1.5 ml, 20.3 mmol) was added to a solution of tert—butyl 2— [(3S,4R){[2-chloro-6—(trifluoromethyl)phenyl]methyl}~3-{[1-(cyclohexen-1— ylmethyl)piperidin—4—yl]carbamoyl}~4—methylpyrrolidin—3-yl]acetate obtained in Example 4i (353 mg, 0.577 mrnol) in dichloromethane (400 pl), which was stirred at room temperature for two hours. The reaction mixture was concentrated under reduced re and purified by ODS column chromatography (elution solvent: methanol/water) to give the title compound (215 mg, yield 67%). 1H—NMR (400 MHZ, CDC13) 8 ppm; 1.01 (3H, d, J=7 Hz), 1.22-1.34 (2H, m), 1.42-1.68 FP11-0417 (7H, m), 1.79-2.07 (6H, m), 2.38-2.46 (1H, m), 2.49 (1H, d, J=10 Hz), 2.56 (1H, d, J=14 Hz), 2.67-2.82 (6H, m), 3.00 (1H, t, J=10 Hz), 3.16 (1H, d, J=10 Hz), 3.63-3.76 (II-I, m), 3.88 (1H, d, J=14 Hz), 4.01 (1H, d, J=14 Hz), 5.55 (1H, s), 7.40 (1H, t, J=8 Hz), 7.64 (1H, d, J=4 Hz), 7.66 (1H, d, J=4 Hz), 8.56 (1H, d, J=8 Hz) Specific on: [(111329 + 37.1 (c 0.11, MeOH) (Example 5: Another method for compound of Example 3) 2—[(3S,4R){[2— Chloro—6—(tn'fluoromethyl)phenyl]methyl}~3—{[1—(cyclohex-1—en—1-y1rnethyl)piperidin—4— yl]carbamoyl}methylpyrrolidin—3~y1]acetic acid (Example 5a) tert—Butyl 4-[(3RS,4SR)[2—(tert—butoxy)~2—oxoethyl]—1~[(2,6- dichlorophenyl)methyl]—4-methylpyrrolidin—3—amido]piperidine—1-carboxylate % palladium hydroxide (300 mg) was added to a solution ofbenzyl (3RS,4SR)— 1-benzy1-3—[2-(tert-butoxy)—2~oxoethyl]—4—methylpyrrolidine—3-carboxylate obtained by a method r to that of Example 1g (3 g, 7.08 mmol) in methanol (200 n11), followed by stirring at room temperature overnight under a hydrogen atmosphere. The on liquid was filtered and washed with methanol. Warm water at about 40°C was added to the product collected by filtration to dissolve the solid, followed by filtration. The filtrate was concentrated together with the first filtrate to give a deprotected form (1.7 g). ol (44 ml), 2,6—dichlorobenzaldehyde (4.83 g, 27.6 mmol) and acetic acid (1.1 ml, 18.4 mmol) were added to the deprotected form obtained by a method similar to the above method (4.47 g, 18.4 rmnol), followed by stirring at room ature for 15 minutes, and sodium triacetoxyborohydride (6.16 g, 27.6 mmol) was added, followed by heating with stirring at 40°C overnight. l buffer [prepared fiom potassium dihydrogenphosphate (13.65 g), disodium hydrogenphosphate dodecahydrate (71.6 g) and water (1.5 1)] was added to the on liquid, ethyl acetate and heptane were added, and the mixture was filtered.

The filtrate was ted with ethyl acetate, the organic layer was dried over sodium sulfate and concentrated, and the ing crude t was then purified by column chromatography (silica gel, elution solvent: methanol/ethyl acetate) and combined with the product collected by filtration to give (3RS,4SR)~3—[2-(tert—butoxy)-2—oxoethyl][(2,6— rophenyl)methyl]—4—methylpyrrolidjnecarboxylic acid (3.9 g). tert—Butyl 4- aminopiperidjne-l—carboxylate (974 mg, 4.89 mmol), N,N-djmethy1formamide (10 ml), triethylamine (1.14 ml, 8.13 mmol) and PyBOP (2.54 g, 4.89 mrnol) were added to this compound (1.64 g, 4.07 mmol), followed by stining at room temperature overnight. Water was added to the reaction liquid, which was extracted with ethyl acetate, and the organic FP11—0417 layer was then washed with water and brine and dried over magnesium sulfate. The crude product ed by concentration was purified by column chromatography (NH silica gel, elution t: ethyl acetate/heptane) to give the title compound (2.10 g, yield: 88.1%). 1H-NMR (400 MHZ, CDCl3) 5 ppm; 0.91 (3H, d, J=7 Hz), 1.15-1.34 (2H, m), 1.40 (9H, s), 1.49 (9H, s), 1.58—1.70 (2H, m), 1.76-1.84 (1H, br), 1.98 (1H, d, J=16 Hz), 2.03-2.13 (1H, m), 2.54—2.65 (2H, m), 2.66-2.88 (2H, br), 2.92 (1H, t, J=10 Hz), 3.11 (1H, d, J=16 Hz), 3.59 (1H, d, J=10 Hz), 3.81—3.90 (1H, m), 3.95”(2H, s), 3.90—4.07 (2H, br), 7.18 (1H, dd, J=7, 8 Hz), 7.33 (1H, d, J=8 Hz), 8.23 (1H, d, J=8 Hz).

Analysis by HPLC; (Analysis conditions) : CHIRALPAK IA actured by Daicel Chemical Industries, Ltd.) (0.46 cm diameter x 15 cm), 40°C, eluent: hexane/ethanol = 95/5 (v/v), flow rate: 1 ml/min., detection: UV (210 nm) (Analysis result) The resulting title nd was analyzed under the above analysis conditions, and a peak with a retention time of 3.46 minutes and a peak with a retention time of6.04 minutes were observed.

(Example 5b) tert-Butyl 4-[(3 S,4R)—3-[2—(tert—butoxy)~2—oxoethyl]—1-[(2,6- dichloropheny1)methy1]—4-methylpyrrolidin—3-amido]piperidine-1—carboxylate tert—Butyl 4—[(3RS,4SR)[2—(tert—butoxy)—2—oxoethyl]—1-[(2,6- dichlorophenyl)methy1]—4—rnethylpyrrolidin~3—arnido]piperidine—1-carboxylate obtained in Example 5a (2.04 g, 3.49 mmol) was optically resolved repeatedly by HPLC (CHRALPAK IA (3 cm diameter x 25 cm), n solvent: hexane/ethanol = 94/6, flow rate: 22 ml/min.) to give the title compound (a chiral form corresponding to the peak with a r retention time) (859 mg, 42.1%) and the optical isomer (a chiral form corresponding to the peak with a longer ion time) (817 mg, 40%), respectively.

Chiral form corresponding to the peak with a shorter retention time; 1H—NMR (400 MHz, CDC13) 5 ppm; 0.91 (3H, d, J=7 Hz), 1.15-1.33 (2H, m), 1.40 (9H, s), 1.49 (9H, s), 1.76-1.84 (1H, m), 1.98 (1H, d, J=16 Hz), 2.01—2.11 (1H, m), 2.57 (1H, d, J=10 Hz), 2.57-2.64 (1H, m), 2.69-2.82 (2H, m), 2.92 (1H, t, J=10 Hz), 3.11 (1H, d, J=16 Hz), 3.59 (1H, d, J=10 Hz), 3.80- 4.09 (2H, m), 3.95 (2H, s), 7.18 (1H, t, J=8 Hz), 7.33 (2H, d, J=8 Hz), 8.23 (1H, d, J=8 Hz).

Analysis by HPLC; (Analysis conditions) Column: CHIRALPAK IA (manufactured by Daioel Chemical Industries, Ltd.) (0.46 cm diameter x 15 cm), 40°C, : hexane/ethanol = 95/5 (V/v), flow rate: 1 m1/min., detection: UV (210 nm) (Analysis ) The resulting chiral form corresponding to the peak with a shorter retention time was analyzed under the above analysis conditions to find that the retention time was 3.44 minutes and the enantiomeric excess was >99% ee.

(Example 50) utyl 4—[(3 S,4R)—3-[2—(tert—butoxy)—2—oxoethyl]—1-{[2—chloro-6— (trifluoromethyl)phenyl]methyl}—4—methylpyrrolidin—3—amido]piperidine—1-carboxylate % palladium hydroxide (75.8 mg) was added to a solution of tert-butyl 4- [(3S,4R)~3-[2-(tert—butoxy)—2—oxoethyl]~1~[(2,6-dichlorophenyl)mefliyl]methylpyrrolidin— 3-amido]piperidine—1-carboxylate obtained in Example 5b (758 mg, 1.29 mmol) in methanol (20 ml), followed by stirring at 40°C for three hours under a hydrogen here. The reaction liquid was filtered, and the filtrate was concentrated to give a crude product (501 mg). 2~(Bromomethy1)—1-chloro—3-(trifluoromethyl)benzene (145 mg, 0.532 mmol) and potassium carbonate (53.2 mg) were added to a solution of this crude product (174 mg) in N,N-dimethylformamide rated) (6 mL), followed by stirring ght. 2— (Bromomethyl)—1—chloro(trifluoromethyl)benzene (145 mg, 0.532 mmol) and potassium carbonate (106 mg) were further added, followed by stirring for four hours. Ethyl acetate was added to the reaction liquid, which was washed with a 1 N aqueous sodium hydroxide solution and brine. This was dried over anhydrous magnesium sulfate and concentrated.

The e was purified by silica gel column tography (elution solvent: heptane/ethyl acetate) to give the title compound (82 mg, yield: 32.4%). 1H—NMR (400 MHz CDC13) 8 ppm; 0.92 (3H, d, J=7 Hz), 1.12-1.18 (2H, m), 1.39 (9H, s), 1.49 (9H, s), 1.53—1.65 (1H, m), 1.76—1.86 (1H, m), 1.98 (1H, d, J=16 Hz), .10 (1H, m), 2.51 (1H, d, J=10 Hz), 2.61-2.81 (3H, m), 2.89 (1H, t, J=10 Hz), 3.12 (1H, d, J=16 Hz), 3.53 (1H, d, J=1O Hz), 3.80—4.10 (5H, m), 7.38 (1H, t, J=8 Hz), 7.63 (2H, d, J=8 Hz), 7.96 (1H, d, J=8 Hz).

MS (ESI) m/z: 618.1 (M+H)+ (Example 5d) 2—[(3S,4R)—1~{[2-Chloro~6-(trifluoromethyl)phenyl]methyl} methyl-3—[(piperidin—4-yl)carbamoyl]pyrrolidin—3-y1]acetic acid Trifluoroacetic acid (3 mL) was added to a solution of tert-butyl 4—[(3S,4R)[2- (tert-butoxy)—2—oxoethyl]— 1-{ [2-chloro(trifluoromethyl)phenyl]methyl} methylpyrrolidin—3—amido]piperidine-l—carboxylate obtained in Example 5c (82 mg, 0.133 mmol) in dichloromethane (dehydrated) (3 mL) under ice—cooling, followed by stirring at room temperature for 2.5 hours. The on liquid was concentrated under reduced pressure, and the residue was purified by ODS column chromatography (elution solvent: FP11-0417 water/memanol) to give the title compound (72 mg, yield: quant.) 1H-NMR (400 MHz, CD3OD) 5 ppm; 0.90 (3H, d, J=5 Hz), .70 (2H, m), .91 (1H, m), 1.99-2.08 (1H, m), 2.12-2.23 (2H, m), 2.61 (1H, d, J=9 Hz), 2.60—2.70 (2H, m), 2.95—3.12 (4H, m), 3.25—3.36 (2H, m), 3.53 (1H, d, J=9 Hz), 3.83—3.92 (1H, m), 3.97 (1H, d, J=13 Hz), 4.07 (1H, d, J=l3 Hz), 7.51 (1H, t, J=8 Hz), 7.73 (1H, d, J=8 Hz), 7.78 (1H, d, J=8 Hz).

MS (ESI) m/z: 462.2 (M+H)+ (Example 5e) 2—[(3S,4R)—1—{[2~Chloro(trifluoromethyl)phenyl]methyl}~3—{[l- (cyclohex—l~en—1—ylmethyl)piperidin—4—yl]carbamoyl}—4—methylpyrrolidin~3—yl]acetic acid ex-l-ene-1—carbaldehyde (25.9 ul, 0.227 mmol), acetic acid (30 ul) and sodium tn'acetoxyborohydride (68.8 mg, 0.325 mmol) were added to a solution of 2- [(3S,4R)-l—{[2—chloro—6-(11ifluoromethyl)phenyl]methyl}—4-methyl[(piperidin4- bamoyl]pyrrolidin—3-yl]acetic acid obtained by the method of Example 5d (15 mg, 0.0325 mrnol) in tetrahydrofiiran (dehydrated) (2 mL), followed by stirring overnight.

Water and methanol were added to the reaction liquid, which was concentrated under reduced pressure, and the residue was purified by ODS column chromatography on solvent: water/methanol) to give the title compound (12.8 mg, yield: 70.8%). 1H—NMR (400 MHz, CD3OD) 8 ppm; 0.89 (3H, d, J=7 Hz), 1.50—1.90 (2H, m), 1.83-1.95 (1H, m), 2.00-2.22 (5H, m), 2.30—2.45 (2H, m), 2.57—2.68 (2H, m), 2.92-3.14 (5H, m), 3.54 (1H, d, J=10 Hz), 3.64-3.78 (1H, m), 3.95 (1H, d, J=13 Hz), 4.05 (1H, d, J=13 Hz), 5.76 (1H, s), 7.49 (1H, t, J=8 Hz), 7.72 (1H, d, J=8 Hz), 7.77 (1H, d, J=8 Hz).

MS (ESI) m/z: 556.3 (M+H)+ (Example 6) 2—[(3S,4R)[(2—Chloro—6~methylphenyl)methyl]~3—{[1-(cyclohex—l- enyhnethyl)piperidin—4—yl]carbamoyl}—4—methylpyrrolidin—3-yl]acetic acid

[0198] (Example 6a) (3S,4R)—3-[2—(tert—Butoxy)—2—oxoethyl]—1-[(2—chloro-6— methylphenyl)methyl]—4—methylpyrrolidine~3~carboxylic acid The title compound (154 mg, yield 27.2%) was ed from (3S,4R)[2—(te1t~ butoxy)-2—oxoethyl]methylpyrrolidine-3~carboxylic acid obtained by a method similar to that ofExample li (360 mg, 1.48 mmol) and 2—chloro-6—methylbenzaldehyde (416 mg, 2.69 mmol) by a method similar to the method described in Example 1j. 1H-NMR (400 MHz, CDC13) 8 ppm; 1.01 (3H, d, J=7 Hz), 1.41 (9H, s), 2.10 (1H, d, J=17 Hz), 2.15223 (1H, m), 2.42 (3H, s), 2.63-2.69 (2H, m), 2.97 (1H, t, J=10 Hz), 3.01 (1H, d, J=17 Hz), 3.68 (1H, d, J=9 Hz), 4.00 (2H, s), 7.09—7.26 (3H, m).

FP11—0417 (Example 6b) tert—Butyl 2-[(3S,4R)—1—[(2—chloro-6—methylphenyl)methyl]~3—{[1- (cyclohex-l-eny1methy1)piperidin—4-y1]carbarnoyl}—4-methy1pyrrolidin—3-yl]acetate The title nd (212 mg, yield 942%) was obtained from (3S,4R)—3-[2-(tert— butoxy)—2—oxoethy1]-1—[(2-chloro—6—methy1phenyl)methy1]—4—methylpyrrolidinecarboxylic acid obtained in Example 6a (154 mg, 0.403 mmol) and 1-(cyclohex—1-en—1— ylmethyl)piperidin—4—amine obtained in Example 4b (110 mg, 0.564 mmol) by a method similar to the method desoribed in Example 1k. 1H—NMR (400 MHz, CDC13) 8 ppm; 0.92 (3H, d, J=7 Hz), 1.21-1.39 (3H, m), 1.40 (9H, s), 1.52-1.69 (5H, m), 1.81-2.12 (8H, m), 1.99 (1H, d, J=16 Hz), 2.44 (3H, s), 2.52 (1H, d, J=10 Hz), 2.61 (1H, dd, J=6, 9 Hz), 2.68 (2H, br d, J=9 Hz), 2.75 (2H, s), 2.86 (1H, t, J=10 Hz), 3.10 (1H, d, J=16 Hz), 3.58 (1H, d, J=10 Hz), 3.60-3.70 (1H, m), 3.79 (1H, d, J=13 Hz), 3.84 (1H, d, J=13 Hz), 5.54 (1H, s), 7.08-7.14 (2H, m), 7.23—7.26 (1H, m), 8.00 (1H, d, J=8 Hz).

(Example 60) 2—[(3S,4R)—1-[(2-Chloro~6-methylphenyl)methyl]—3-{[1-(cyclohex— l-en-l -ylmethyl)piperidin—4—y1]carbamoyl}~4-methy1pyrrolidin—3-y1]acetic acid The title compound (140 mg, yield 73.4%) was ed from utyl 2— [(3S,4R)- chloro-6—methylphenyl)methy1]—3—{ [1 ohexen-1 -y1methyl)piperidin— 4-y1]carbamoyl}—4-methy1pyrrolidin—3-y1]acetate obtained in Example 6b (212 mg, 0.38 mmol) by a method similar to the method described in Example 1m. 1H—NMR (400 MHz, CD30D) 8 ppm; 0.93 (3H, d, J==7 Hz), 1.57—1.80 (6H, m), 1.90-2.39 (8H, m), 2.49 (3H, s), 2.61-3.38 (8H, m), 3.59—4.20 (4H, m), 5.95 (1H, s), 7.18-7.39 (3H, m).

(Example 7) ,4R)—1~{[2—Chloro—6-(t1ifluoromethyl)phenyl]methyl}{[1- (cyclopent—l—en—l -y1methy1)piperidin—4-yl]carbamoyl}—4-methylpyrrolidin—3-yl]acetic acid (Example 7a) Cyclopent—l—ene—l-carbaldehyde Amixture of sodium periodate (28.6 g, 134 mrnol) and water (250 ml) was added to a mixture of 1,2-cyclohexanediol (12 g, 103 mmol) and diethyl ether (150 ml), which was stirred at room temperature for 35 minutes. A20% aqueous potassium hydroxide solution (40 ml, 206 mmol) was added to the reaction e, which was stirred at room temperature for two hours. The on mixture was extracted with diethyl ether twice. The organic layer was dried over anhydrous magnesium sulfate, and the solvent was distilled offto give the title compound (6.1 g, yield 61.6%). 1H-NMR (400 MHZ, CDC13) 5 ppm; 1.96-2.04 (2H, m), 2.50-2.57 (2H, m), 2.58-2.66 (2H, m), 6.87-6.90 (1H, m), 9.80 (1H, s).

(Example 7b) 2—[(3S,4R)—1—{[2—Chloro(trifluoromethyl)pheny1]methy1}-3~{[1- FP11—0417 (cyclopent—1~enylmethyl)piperidin—4—yl]carbamoyl}—4~methylpyrrolidin—3-yl]acetic acid The title compound (288 mg, yield: 73.5%) was obtained from 2-[(3S,4R)-1—{[2- chloro(uifluoromethyl)pheny1]methyl}methyl—3-[(piperidjn yl)carbamoyl]pyrrolidin—3~y1]acetic acid obtained in e 3d (334 mg, 0.723 mmol), cyclopent-l-ene—l-carbaldehyde obtained in Example 7a (209 mg, 2.17 mmol), acetic acid (300 pd) and sodium triacetoxyborohydride (460 mg, 2.17 mmol) by a method similar to the method described in Example 3e. 1H—NMR (400 MHZ, CD30D) 5 ppm; 0.89 (3H, d, J=7 Hz), 1.24-1.35 (2H, m), .66 (2H, m), 1.70-1.78 (1H, m), 1.85-1.99 (3H, m), .22 (2H, m), 2.29—2.42 (4H, m), 2.61 (1H, d, J=10 Hz), 2.60-2.68 (1H, m), 2.91—3.02 (2H, m), 3.07 (1H, ’d, J=1'6 Hz), 3.54 '(1H, d, J=10 Hz), 3.64—3.78 (1H, m), 3.95 (1H, d, J=l4 Hz), 4.05 (1H, d, J=14 Hz), 5.74 (1H, s), 7.50 (1H, t, J=8 Hz), 7.71 (1H, d, J=8 Hz), 7.78 (1H, d, J=8 Hz).

MS (ESI) m/z: 564.3 (M+Na)+ (Example 8) 2-[(3S,4R)~1-[(2~Chloro~6—methylphenyl)methyl]~3-{[(l—cyclopent-l- enylmethyl)piperidin—4—yl]carbamoyl}—4-mefl1ylpyrrolidin—3—yl]acetic acid (Example 8a) tert—Butyl 4-[(3S,4R)—3-[(2—tert—butoxy)—2—oxoethyl]—l-[(2—chloro-6— methylphenyl)methyl]methylpyrrolidin—3~amido}piperidine-l-carboxylate The title compound (237 mg, yield 91.7%) was obtained fiom (3S,4R)-3—[(2-te1t— butoxy)—2—oxoethyl][(2-chloro—6—methylphenyl)methyl]—4-methylpyrrolidine~3—carboxylic acid ed by a method r to the method described in Example 6a (175 mg, 0.458 mmol) and tert—butyl 4-aminopipe1idine~1—carboxylate (137 mg, 0.684 mmol) by a method similar to the method described in Example 1k. 1H—NMR (400 MHZ, CDC13) 8 ppm; 0.92 (3H, d, J=8 Hz), 1.15-1.30 (2H, m), 1.40 (9H, s), 1.49 (9H, s), 1.58-1.68 (2H, m), 1.78-1.88 (1H, br), 1.99 (1H, d, J=8 Hz), 2.05—2.11 (1H, m), 2.42 (3H, s), 2.50 (1H, d, J=10 Hz), 2.60—2.66 (1H, m), 2.70-2.81 (2H, br), 2.85 (1H, t, J=10 Hz), 3.10 (1H, d, J=16 Hz), 3.55 (1H, d, J=10 Hz), 3.74-4.10 (4H, m), 7.07-7.15 (2H, m), 7.22-7.27 (1H, m), 8.14 (1H, d, J=8 Hz).

(Example 8b) 2—[(3S,4R)—1-[(2—Chloro—6—methylpheny1)methyl]—3-{[(l-cyclopent— 1—en-l-ylmethyl)piperidin—4~yl]carbamoyl}methylpyrrolidin—3—y1]acetic acid A mixture of tert-butyl 4-[(3S,4R)-3—[(2—tert—butoxy)-2—oxoethyl]—1-[(2-chloro methylphenyl)methyl]—4-methylpyrrolidin—3-amido}piperidine~1-carboxylate obtained in Example 8a (237 mg, 0.42 mmol), m'fluoroacetic acid (2.3 ml, 31 mmol) and romethane (dehydrated) (2.3 ml) was stiired at room temperature for three hours and FP11—0417 minutes. The reaction mixture was concentrated, and ent—l-ene—l—carbaldehyde obtained in Example 7a (121 mg, 1.26 mmol), triethylamine (0.176 ml, 1.26 mmol) and tetrahydrofiiran (5 ml) were added to the g residue, followed by stirring for 35 minutes. Sodium tn'acetoxyborohydride (267 mg, 1.26 mmol) was added to the reaction mixture, which was d at room temperature for 13 hours. The reaction mixture was concentrated, and the resulting residue was purified by silica gel column tography (ODS silica gel, elution solvent: water/methanol) twice to give the title compound (88 mg, yield 42.9%). 1H—NlVLR (400 MHZ, CD30D) 8 ppm; 0.92 (3H, d, J=7 Hz), 1.50-1.68 (2H, br), .83 1‘0 (1H, br), 1.88—2.00 (3H, m), 2.19-2.30 (2H, m), 2.31—2.43 (6H, m), 2.48 ‘(3H, s), 2.62-2.72 (2H, m), 2.90—3.09 (4H, m), 3.25—3.40 (2H, m), 3.61 (1H, d, J=10 Hz), 3.66-3.78 (1H, m), 3.89-4.06 (2H, m), 5.73 (1H, s), 7.14-7.20 (2H, m), 7.25—7.30 (1H, m).

(Example 9) 2—[(3S,4R)—3-{[(3S)—1-(Cyclohex—1—eny1methyl)pyrrolidin—3- yl]carbamoyl}~l—[(2,6-dichlorophenyl)methyl]—4—methylpyrrolidin—3—y1]acetic acid

[0208] (Example 9a) tert—Butyl N—[(3S)(cyclohex—1-eny1methyl)py1rolidin—3- yl]carbamate l-Cyclohexene-l-carboxaldehyde (1.48 ml, 13 mmol) was added to a mixture of (3S)—(—)—3~(tert—butoxycarbonylamino)pyrrolidine (2.0 g, 10.8 mmol), acetic acid (1.24 ml, 21.6 mmol) and tetrahydrofiiran (dehydrated) (27 ml), ed by stirring at room temperature for 15 minutes, and sodium triacetoxyborohydn'de (4.58 g, 21.6 mmol) was then added and the reaction mixture was stirred at room ature for 17 hours and 45 minutes.

A ted aqueous sodium bicarbonate solution was added to the reaction mixture, which was extracted with ethyl acetate. The c layer was dried over sodium sulfate, and the solvent was distilled 011‘. The residue was purified by NH silica gel column chromatography (elution solvent: ethyl acetate/heptane) to give the title compound (1.3 g, yield: 42.9%). , 1H—NMR (400 MHZ, CDC13) 6 ppm; 1.44 (9H, s), 1.50-1.65 (5H, m), 1.95-2.03 (4H, br), 2.17-2.29 (2H, m), 2.40-2.55 (2H, m), 2.65-2.74 (1H, m), 2.84-2.94 (2H, m), 4.10—4.19 (1H, br), 4.77-4.87 (1H, br), 5.56 (1H, br s).

[0209] (Example 9b) (3S)(Cyclohex—1-en—1-y1methy1)pyrrolidin—3-amine A mixture of tert—butyl N—[(3S)(cyclohexen—1~ylmethyl)py1rolidin—3- yl]carbamate obtained in Example 9a (1.3 g, 4.64 mmol) and ethanol (9.2 ml) was stirred under ice-cooling, a 5 N aqueous hloric acid solution (9.28 ml, 46.4 mmol) was FP11—0417 added, and the reaction mixture was stirred at room temperature for 14 hours and 30 minutes.

The reaction mixture was stirred under ice-cooling, a 5 N aqueous sodium hydroxide solution (9.28 ml, 46.4 mmol) was added, and the solvent was distilled off. Ethanol was added to the residue, the precipitated solid was filtered 011°, and the solvent in the filtrate was distilled off. Ethanol was fiirther added to the residue, the precipitated solid was filtered off, and the t in the filtrate was distilled 011"to give the title compound (0.8 g, yield 95.6%). 1H-NTVIR (400 MHZ, CD3OD) 5 ppm; 1.50—1.68 (6H, m), .05 (3H, m), 2.14-2.23’(1H, m), 2.28—2.32 (1H, m), 2.43-2.49 (1H, m), 2.63-2.73 (2H, m), 2.91-2.98 (2H, m), 3.44—3.50 (1H, m), 5.62 (1H, br s).

[0210] (Example 9c) tert—Butyl 2-[(3S,4R){[(3S)—1~(Cyclohex—l-en—1— ylmethyl)pyrrolidin—3-yl]carbamoyl}~l~[(2,6~dichlorophenyl)methyl]~4—methylpyrrolidin—3- yl]acetate The title compound (236 mg, yield 58%) was obtained from (3S)—1—(cyclohex—l- en—l-yhnethyl)pyrrolidin—3-amine obtained in e 9b (169 mg, 0.94 mmol) and (3S,4R)[2—(tert-butoxy)-2—oxoethyl]-1—[(2,6—dichlorophenyl)methyl]—4-methylpyrrolidine— oxylic acid obtained in Example 2k (290 mg, 0.72 mmol) by a method similar to the method described in e 1k. 1H—NMR (400 MHZ, CDC13) 6 ppm; 0.91 (3H, d, J=7 Hz), 1.41 (9H, s), .63 (6H, m), 1.87—2.14 (6H, m), 2.20-2.25 (1H, m), 2.33-2.40 (1H, m), .67 (4H, m), 2.79-2.94 (3H, m), 3.11 (1H, d, J==16 Hz), 3.62 (1H, d, J=10 Hz), 3.92-4.02 (2H, m), 4.28—4.38 (1H, In), 5.74 (1H, br s), 7.17 (1H, dd, J=7, 8 Hz), 7.32 (2H, d, J=8 Hz), 8.30 (1H, d, J=7 Hz).

(Example 9d) 2-[(3S,4R)—3~{[(3S)—l—(Cyclohex—1-en-1—ylmethyl)pyrrolidin—3— yl]carbamoyl}—1—[(2,6—diclflorophenyl)methyl]—4-methylpyrrolidin—3~y1]acetic acid The title compound (173 mg, yield 81%) was obtained fiom tert—butyl 2-[(3S,4R)— 3-{[(3S)(cyclohex—l-en—1—yhnethyl)pyrrolidin—3-yl]carbamoyl}—1-[(2,6— dichlorophenyl)methyl]~4—methylpyrrolidin—3-yl]acetate obtained in Example 90 (236 mg, 0.42 mmol) by a method similar to the method descfibed in Example 1m. 1H—NMR (400 MHZ, CD3OD) 5 ppm; 0.89 (3H, d, J=7 Hz), 1.55—1.67 (4H, m), 1.73—1.84 (1H, m), 1.92-2.32 (7H, m), 2.58-2.68 (2H, m), 2.74-2.85 (2H, m), .15 (4H, m), 3.20- 3.34 (2H, m), 3.61 (1H, d, J=10 Hz), 4.00-4.08 (2H, m), .27 (1H, m), 5.74 (1H, br s), 7.29 (1H, dd, J=7, 9 Hz), 7.40-7.44 (2H, m).

(Example 10) 2-[(3S,4R){[2-Chloro(difluoromethyl)phenyl]methyl}[(l- hexylpiperidin—4—yl)carbam0yl]—4-methylpy1rolidin—3—yl]acetic acid FP11~0417 (Example 10a) l-Hexylpiperidin—4—arnine The title compound was obtained by a method r to the method described in U82005/0222175 A1 and Examples 4a and 4b. 1H—NMR (400 MHz, CDCl3) 5 ppm: 0.80 (3H, t, J=7 Hz), 1.05—1.45 (12H, m), 1.73 (2H, d, J=6 Hz), 1.88 (2H, t, J=6 Hz), 2.21 (2H, dd, J=6, 8 Hz), 2.52-2.62 (1H, m), 2.79 (2H, d, J=12 (Example 1%) Methyl 3-chloro-2—methy1benzoate Iodomethane (1.96 ml, 31.5 mmol) was added to a mixture of 3—chloro—2— methylbenzoic acid (3.58 g, 21 mmol), potassium carbonate (5.8 g, 42 mmol) and N,N- dimethylformarnide (35.9 ml), followed by stirring at room temperature for 18 “hours and 30 minutes. Water and ethyl acetate were added to the reaction liquid, and the organic layer was extracted. The organic layer was sequentially washed with a saturated aqueous ammonium chloride solution and brine, and then dried over sodium sulfate, filtered and concentrated. The residue was purified by silica gel column tography (elution solvent: heptane/ethyl acetate) to give the title compound (3.67 g, yield: 97%). 1H—NMR (400 lVH-Iz, CDC13) 5 ppm; 2.60 (3H, s), 3.91 (3H, s), 7.13-7.21 (1H, m), 7.50 (1H, dd, J=1, 8 Hz), 7.70 (1H, dd, J=1, 8 Hz).

(Example 10c) Methyl 2—(bromomethyl)—3-chlorobenzoate A suspension ofmethyl 3-chloro—2—methylbenzoate obtained in Example 10b (1 g, 5.42 mmol), carbon tetrachloride (13.3 ml), N—bromosuccinimide (1.06 g, 5.96 mrnol) and benzoyl peroxide (3.5 mg, 0.0108 mmol) was heated in a 90°C oil bath under a nitrogen stream. After three hours and 45 minutes, heating was completed, and the mixture was diluted with water, ethyl e and a ted aqueous sodium bicarbonate solution. The organic layer was washed with a saturated aqueous sodium bicarbonate solution, dried over ous magnesium sulfate, d and concentrated to give the title compound (1.43 g, yield: 100%). 1H—NMR (400 MHz, CDClg) 5 ppm; 4.00 (3H, s), 5.12 (2H, s), 7.32 (1H, t, J=8 Hz), 7.58 (1H, dd, J=2, 8 Hz), 7.86 (1H, dd, J=2, 8 Hz).

(Example 10d) [2—(Bromomethyl)—3—chlorophenyl]methanol Dichloromethane (10 ml) was added to methyl 2—(bromomethyl)—3-chlorobenzoate obtained in Example 100 (500 mg, 1.9 mmol), a 1.04 M diisobutylalurninum hydride/n- hexane solution (4.57 ml, 4.75 mmol) was added at -78°C, ed by stirring for one hour under a nitrogen here. A saturated aqueous Rochelle salt on and tert—butyl FP11-0417 methyl ether were added, followed by extraction with tert—butyl methyl ether. The organic layer was washed with brine, dried over magnesium sulfate and then allowed to pass through a silica gel pad. The eluate was concentrated to give the title compound (440 mg, yield 98.3%). 1H—NMR (400 MHz, CDC13) 5 ppm; 1.82 (1H, t, J=6 Hz), 4.78 (2H, s), 4.85 (2H, d, J=5 Hz), 7.25-7.28 (1H, m), 7.31-7.40 (2H, m).

"(Example 10e) momethyl)~3~chlorobenzaldehyde Dichloromethane (15 ml) and ese dioxide (1.69 g, 19.4 mmol) were added to [2-(bromomethyl)chlorophenyl]methanol ed in Example 10d (440 mg, 1.87 1'0 mmol), followed by stirring at room temperature overnight. Manganese dioxide (1.2 g, 13.8 mmol) was fiirther added, followed by heating with stirring at 40°C for 2.5 hours. The reaction mixture was filtered through Celite and washed with ethyl acetate. The e was concentrated under reduced re, and the resulting crude product was purified by column chromatography (silica gel, elution solvent: heptane/ethyl acetate = 99/1 ——> 90/10) to give the title compound (289 mg, yield 66.2%). 1H—NMR (400 MHz, CDC13) 5 ppm; 5.13 (2H, s), 7.48 (1H, t, J=8 Hz), 7.66 (1H, dd, J=l, 8 Hz), 7.77 (1H, dd, J=1, 8 Hz), 10.23 (1H, s).

(Example 10f) 2-(Bromomethyl)—l-chloro-3—(difluoromethyl)benzene Dichloromethane (10 ml) was added to 2—(bromomethyl)—3—chlorobenzaldehyde obtained in Example lOe (289 mg, 1.24 mmol), [bis(2-methoxyethyl)amino]sulfi1r ride (457 pl, 2.48 mmol) was added at 0°C, ed by ng at room ature for 1.5 hours under a nitrogen atmosphere. A saturated aqueous sodium bicarbonate solution was added at 0°C to the reaction mixture, which was extracted with diethyl ether. The organic layer was washed with brine, dried over magnesium sulfate and then concentrated. The resulting crude product was purified by column chromatography (silica gel, elution solvent: heptane/ethyl acetate = 99/1 ——-> 95/5) to give the title compound (238.3 mg, yield 75.2%). 1H—NMR (400 MHz, CDC13) 5 ppm; 4.74 (2H, s), 6.93 (1H, t, J=55 Hz), 7.37 (1H, t, J=8 Hz), 7.53 (2H, t, J=8 Hz).

(Example 10g) (3RS,4SR)[2-(tert-Butoxy)-2~oxoethyl]-l-[(2,6- dichlorophenyl)methyl]—4-methy1pyrrolidine—3-carboxy1ic acid % palladium hydroxide (500 mg) was added to a solution ofbenzyl (3RS,4SR)- 1-benzyl[2-(te1t—butoxy)-2—oxoethyl]—4-methylpyrrolidinecarboxy1ate obtained by a method r to that ofExample 1g (8.35 g, 19.7 mmol) in methanol (200 ml), followed by FP11-0417 stirring at room temperature overnight under a hydrogen atmosphere. The reaction mixture was filtered, and palladium on the filter paper was washed well with hot water at 50°C. The e was concentrated and azeotropically distilled with methanol and toluene well to give a white solid (3.92 g). Methanol (20 ml), chlorobenzaldehyde (5.64 g, 32.2 mmol), acetic acid (966 pl, 16.1 mmol) and sodium triacetoxyborohydn'de (6.82 g,32.2 mmol) were added thereto, followed by stirring at room temperature overnight. The reaction mixture was fiirther heated with stining in a 50°C hot water bath for three hours. Neutral bufi‘er [prepared fiom ium dihydrogenphosphate (13.65 g), disodium hydrogenphosphate dodecahydrate (71.6 g) and water (1.5 L)] and ethyl acetate were added to the reaction mixture, and the solid was filtered ofi‘. The solid on the filter paper was washed with ethyl acetate to give the title nd as a white solid. The filtrate was extracted with ethyl acetate, and the organic layer was then dried over sodium sulfate, concentrated, purified by column tography (silica gel, elution solvent: methanol/ethyl acetate) and combined with the solid on the filter paper to give the title compound (3.77 g, yield 58.2%). 1H—NMR (400 MHZ, CDC13) 5 ppm; 1.01 (3H, d, J=7 Hz), 1.42 (9H, s), 2.09 (1H, d, J=16 Hz), 2.14-2.24 (1H, m), 2.62—2.69 (2H, m), 2.99 (1H, t, J=10 Hz), 3.02 (1H, d, J=10 Hz), 3.71 (1H, d, J=10 Hz), 4.08 (1H, d, J=12 Hz), 4.12 (1H, d, J=12 Hz), 7.21 (1H, d, J=8 Hz), 7.23 (1H, d, J=8 Hz), 7.37 (1H, d, J=8 Hz).

(Example 10h) tert—Butyl 2—[(3 S,4R)—l-[(2,6—dichlorophenyl)methyl]~3-[(1- hexylpiperidin—4—yl)carbamoyl]—4-methylpyrrolidin—3-yl]acetate 1-Hexy1piperidin-4—amine obtained in Example 10a (894 mg, 4.85 mmol), tn'ethylamine (1.04 ml, 7.46 mmol), N,N~dimethylformamide (10 ml) and PyBOP (2.52 g, 4.85 mmol) were added to (3RS,4SR)[2—(tert—butoxy)-2—oxoethyl]—1{(2,6- dichlorophenyl)methyl]methylpyrrolidine—3—carboxylic acid obtained in Example 10g (1.5 g, 3.73 mmol), followed by stirring at room temperature ght. Water was added to the reaction mixture, which was extracted with ethyl acetate. The organic layer was washed with water and brine and dried over sodium sulfate. This was trated, and the crude t was purified by column chromatography (NH silica gel, elution solvent: ethyl acetate/heptane) to give a white solid (1.85 g). This was optically ed by HPLC LPAK IA (3 cm diameter x 25 cm), elution solvent: ethanol/hexane = 6/94, flow rate: 20 ml/min.) to give a chiral form corresponding to the peak with a shorter retention time (875 mg).

Analysis by HPLC; FP11-0417 (Analysis ions) Column: PAK IA (manufactured by Daicel Chemical Industries, Ltd.) (0.46 cm diameter x 15 cm), 40°C, eluent: hexane/ethanol = 9/1 (v/V), flow rate: 1 ml/min., detection: UV (210 nm) (Analysis result) The resulting chiral form was analyzed under the above analysis conditions to find that the retention time was 4.85 minutes and the omeric excess was >99% ee.

(Example 10i) utyl 2—[(3S,4R)[(1—hexylpiperidin—4—yl)carbamoyl]—4— methylpyxrolidin—B—ylkcetate Methanol (10 ml) and 20% palladium hydroxide (50 mg) were added to 250 mg of tert-butyl 2-[(3S,4R)—1-[(2,6-dichlorophenyl)methyl][(1—hexylpiperidin—4—yl)carbamoyl]— 4—methylpyrrolidin-3—yl]aeetate obtained in Example l’Oh, f0110wed ”by stirring at room temperature for two hours under a hydrogen atmosphere. The reaction e was filtered, and the filtrate was concentrated to give the title compound (180 mg). 1H—NMR (400 MHz, CDC13) 5 ppm; 0.85—0.90 (3H, m), 1.04 (3H, d, J=6 Hz), 1.25-1.35 (5H, m), 1.42 (9H, s), 1.78-2.10 (6H, m), 2.26-2.50 (4H, m), .05 (3H, m), 3.23 (1H, d, J=13 Hz), 3.29-3.54 (5H, m), 3.55-3.67 (1H, m), 4.17 (1H, d, J=13 Hz), 4.27—4.40 (1H, br), 7.82-7.92 (1H, br).

(Example 103') tert—Butyl 2-[(3 S,4R)—1-{[2~chloro—6- (difluoromethyl)phenyl]methyl}[(l-hexylpiperidin-4—yl)carbamoyl]-4~methylpyrrolidin— 3-yl]acetate 2O N,N—Dimethylformamide (400 pl), momethyl)—1—chloro—3- (difluoromethyl)benzene obtained in Example 10f (24.9 mg, 0.0976 mmol) and potassium carbonate (20.2 mg, 0.146 mmol) were added to utyl 2—[(3S,4R)—3-[(1-hexylpiperidin- 4—yl)carbamoy1]~4—methylpyrrolidin—3-yl]aeetate obtained in Example mi (20 mg, 0.0488 mmol), followed by stirring at room temperature overnight. Ethyl acetate and water were added to the reaction mixture, which was extracted with ethyl acetate. The organic layer was washed with brine and dried over magnesium sulfate. This was concentrated, and the resulting crude product was purified by column chromatography (NH silica gel, elution t: heptane/ethyl acetate = 98/2 —~> 80/20) to give the title compound (14.4 mg, yield 50.2%). 1H—NMR (400 MHz, CDC13) 5 ppm; 0.87-0.94 (6H, m), 1.22-1.35 (6H, m), 1.40 (9H, s), 1.40-1.49 (2H, m), 1.68-1.76 (1H, m), .12 (7H, m), 2.24-2.28 (2H, m), 2.54 (1H, d, J=1O Hz), 2.59-2.63 (1H, m), 2.74-2.88 (3H, m), 3.13 (1H, d, J=16 Hz), 3.61 (1H, d, J=10 Hz), 3.61-3.72 (1H, m), 3.91 (1H, (1, 1:13 Hz), 3.95 (1H, d, J=13 Hz), 6.96 (1H, t, J=55 Hz), FP11—0417 7.35 (1H, t, J=8 Hz), 7.54 (2H, d, J=8 Hz), 7.65 (1H, d, J=8 Hz).

(Example 10k) 2-[(3S,4R)-l-{[2-Chloro-6—(difluoromethyl)phenyl]methyl}[(l- hexylpiperidin—4-yl)carbamoyl]—4-methylpyrrolidin—3-yl]acetic acid The title compound (13.9 mg, yield 61.5%) was obtained fiom tert—butyl 2- [(3S,4R)—1-{[2-chloro-6—(difluoromethyl)phenyl]methy1}—3-[(l-hexylpiperidin—4— yl)carbamoyl]—4—methylpyrrolidin-3—yl]acetate obtained by a method r to that of Example 10j (25 mg, 0.0428 mmol) by a method similar to the method described in Example 1m. 1H—NMR (400 MHz, CDC13) 5 ppm; 0.82090 (3H, m), 0.93 (3H, d, J=7 Hz), 1.23-1.34 1'0 (6H, m), .64 (2H, rn), 1.76—1198“(4H, In), 2.22-2.44 '(4H, m), 2.50—2.78 (SH, m), 2.91 (1H, t, J=8 Hz), 3.22—3.35 (1H, br), .43 (1H, m), 3.70-3.82 (1H, m), 3.92 (1H, d, J=14 Hz), 3.96 (1H, d, J=l4 Hz), 7.21 (1H, t, J=55 Hz), 7.30 (1H, t, J=8 Hz), 7.48 (1H, d, J=8 Hz), 7.55 (1H, d, J=8 Hz), 8.92-9.08 (1H, br). le 11) 2—[(3S,4R){[(l—Cyclohex-l—en-l~ylmethyl)piperidin—4— yl]carbamoyl}-1~[(2,6—dichlorophenyl)methyl]methylpyrr01idin—3-yl]acetic acid (Example 11a) (3S,4R)—3-[2-(tert-Butoxy)—2—oxoethyl]—1-[(2,6— dichlorophenyl)methyl]-4—methylpyrrolidine—3-carboxylic acid Methanol (3 ml), 2,6—dichlorobenzaldehyde (431 mg, 2.46 mmol), acetic acid (73.8 r11, 1.23 mmol) and sodium triacetoxyborohydride (521 mg, 2.46 mmol) were added to (3S,4R)—3-[2—(tert—butoxy)-2—oxoethyl]—4—methylpyrrolidine—3—carboxylic acid obtained by a method similar to that of Example 1i (300 mg, 1.23 mmol), followed by stirring at room temperature for two days. Water was added to the system, which was extracted with ethyl acetate, and the c layer was dried over sodium sulfate and concentrated. The ing crude product was purified by column chromatography (silica gel, elution solvent: methanol/ethyl acetate) to give the title compound (180 mg). On the other hand, the aqueous layer upon partitioning was concentrated and purified by ODS column chromatography (elution solvent, methanol/water) to recover )[2-(tert~butoxy)-2— oxoethyl]—4—methylpyrrolidinecarboxylic acid (120 mg). "Ihe title compound (42.7 mg) was obtained by performing similar reaction again using the red raw material, and was combined with the first title compound (180 mg). 1H—NMR (400 MHZ, CDC13) was confirmed to be identical to that ofthe compound obtained in Example 10g.

(Example 11b) tert—Butyl 2—[(3S,4R)-{3-[(1—cyclohex—l—en—l—ylmethyl)pipe1idin—4— FP11—0417 yl]carbamoyl}-1 -[(2,6—dichlorophenyl)methyl]—4—methylpyrr01idinyl]acetate The title compound (274 mg, yield 85.8%) was obtained from (3S,4R)[2—(tert— butoxy)oxoethyl]—1-[(2,6-dichlorophenyl)methyl]methy1pyrrolidinecarboxy]ic acid obtained in Example lla (222 mg, 0.55 mmol) and l-(cyclohex—1~enylmethyl)piperidin~ 4-amine obtained in Example 4b (150 mg, 0.772 mmol) by a method similar to the method described in Example 1k.

/IR‘(400 DCl3) 5 ppm; 0.91 (3H, d, J=8 Hz), 1.31-1.50 (2H, m), 1.42 (9H, s), 1.52-1.72 (5H, m), 1.81—2.12 (9H, m), 2.55-2.70 (4H, m), 2.75 (2H, s), 2.92 (1H, t, J=10 Hz), 3.12 (1H, d, J=l6 Hz), 3.60-3.72 (2H, m), 3.96 (2H, s), 5.54 (1H, s), 7.18 (1H, t, J=8 Hz), 7.33 (2H, d, J=8 Hz), 8.09 (1H, d, J=8 Hz).

(Example 110) 2-[(38,4R){[(l-Cyclohex-l—en—l-ylmethyl)piperidin bamoy1}-l-[(2,6-dichlorophenyl)methyl]—4-methylpyrrolidin~3-y1]acetic acid The title nd (169 mg, yield 68.2%) was obtained from tert—butyl 2— [(3S,4R)—{3~[(1~cyclohex-l-enylmethy1)piperidin—4-y1]carbamoyl}[(2,6- dichlorophenyl)methyl]-4—methy1pyrrolidin—3-yl]acetate obtained in Example 11b (274 mg, 0.474 mmol) by a method similar to the method described in Example 1m. 1H—NMR (400 MHZ, CD30D) 5 ppm; 0.89 (3H, d, J=7 Hz), 1.56-1.75 (6H, m), 1.78-1.94 (2H, m), 2.00-2.26 (6H, m), 2.36—2.48 (2H, m), 2.60-2.64 (1H, m), 2.68 (1H, d, J=10 Hz), 2.95-3.19 (6H, m), 3.62 (1H, d, J=10 Hz), 3.68-3.80 (1H, br), 3.99—4.07 (2H, m), 5.76 (1H, . s), 7.29 (1H, t, J=8 Hz), 7.43 (2H, d, J=8 Hz).

(Example 12) ,4R)—1—{[2—Chloro(difluoromethyl)phenyl]methyl}-3—{[1— (cyclohex~1~en—1~ylmethyl)pipe1idin—4~yl]carbamoyl}—4-methylpyrrolidin—3—yl]acetic acid (Example 12a) tert—Butyl 4—[(3S,4R)[2-(tert—butoxy)~2~oxoethyl]~1-{[2—chloro— 6~(difluoromethyl)pheny1]methyl}—4-methylpyrrolidin~3~amido]piperidine—1-carboxylate tert-Butyl 4—[(3S,4R)—3—[2—(tert—butoxy)—2-oxoethyl]—4—methylpyrrolidin—3- amido]piperidine-l-carboxylate obtained in Example 3b (375 mg, 0.881 mmol) was ved in N,N-dimethylformamide (7 ml). Potassium carbonate (304 mg, 2.2 mmol) and 2—(bromomethy1)chloro(difluoromethyl)benzene obtained by a method similar to that ofExample 10f(450 mg, 1.76 mmol) were added thereto, and the mixture was heated in a 45°C oil bath for two hours. The reaction e was partitioned between ethyl acetate and a ted aqueous ammonium chloride solution. The ted organic layer was washed with a saturated aqueous ammonium chloride solution three times, dried over anhydrous magnesium sulfate, filtered and concentrated The residue was purified by silica FP11—0417 gel column chromatography (silica gel, elution solvent: heptane/ethyl acetate) to give the title compound (332 mg, yield: 62.8%). 1H—NMR (400 MHz, CDC13) 6 ppm; 0.92 (3H, d, J=7 Hz), 1.13-1.74 (3H, m), 1.39 (9H, s), 1.49 (9H, s), 1.75—1.87 (1H, m), 2.00 (1H, d, J=16 Hz), 2.04-2.15 (1H, m), 2.52 (1H, d, J=10 Hz), 2.56-2.83 (3H, m), 2.86 (1H, t, J=10 Hz), 3.12 (1H, d, J=10 Hz), 3.57 (1H, d, J=10 Hz), 3.80—4.16 (3H, m), 3.90 (1H, d, J=13 Hz), 3.96 (1H, d, J=13 Hz), 6.93 (1H, t, J=55 Hz), 7.37 (1H, t, J=8 Hz), 7.55 (2H, d, J=8 Hz), 7.77 (1H, d, J=8 Hz).

(Example 12b) 2—[(3S,4R)—1-{[2-Chloro(difluoromethyl)phenyl]metl'ryl}{[1- (cyclohex—l~en—l—ylmethy1)piperidin~4—yl]carbamoyl}—4~methylpyrrolidin—3—yl]acetic acid 1‘0 tert-Butyl 4-‘[(3‘S,4R)—3-’[2—(tert~but0xy)oxoethyl]-1—{[2—chloro46- (difluoromethyl)phenyl]methyl}—4—methylpyrrolidin~3-amido]piperidine—1-carboxylate obtained in Example 12a (332 mg, 0.553 mmol) was dissolved in romethane (3.0 ml), trifluoroaoetic acid (3 .0 ml) was added thereto, and the mixture was then left to stand at room temperature. After two hours and 30 minutes, the reaction liquid was concentrated and azeotropically distilled with dichloromethane twice to give an ediate. 186 mg of the resulting intermediate (372 mg) was dissolved in ydrofuran (5 ml), ylamine (116 141, 0.831 mmol) and cyclohex—l-ene-l-carbaldehyde (91.5 mg, 0.831 mmol) were added, and the mixture was left to stand at room temperature. After 30 minutes, sodium tn'acetoxyborohydride (176 mg, 0.831 mmol) was added thereto, and the mixture was d at room temperature. After 10 hours and 50 minutes, the mixture was concentrated. The residue was purified by silica gel column chromatography (ODS, elution solvent: water/methanol) to give the title compound (80 mg) as a white solid. 1H—NMR (400 MHz, CD3OD) 8 ppm; 0.90 (3H, d, J=7 Hz), 1.57—1.75 (6H, m), 1.83—1.96 (2H, m), 2.02-2.13 (4H, m), 2.15-2.27 (1H, m), 2.27 (1H, d, J=16 Hz), 2.46-2.65 (3H, m), 2.61 (1H, d, J=10 Hz), 2.95 (1H, t, J=9 Hz), 2.97 (1H, d, J=16 Hz), 3.03-3.16 (2H, m), 3.20- 3.27 (2H, m), 3.50 (1H, d, J=10 Hz), 3.74-3.83 (1H, m), 3.98 (1H, d, J=13 Hz), 4.02 (1H, d, J=13 Hz), 5.78-5.86 (1H, br), 7.26 (1H, t, J=55 Hz), 7.42 (1H, t, J=8 Hz), 7.60 (2H, d, J=8 Hz).

(Example 13) 2-[(3S,4R)—1-{[2—Chloro(difluoromethyl)phenyl]methyl} {[l- 3O (cyclopent—l -en—1 -ylmethyl)piperidin—4—yl]carbamoyl}methylpyrrolidin—3-yl]acetic acid The title compound (76 mg) was obtained by a method similar to the method described in e 12b using the intermediate ed in e 12b (186 mg) and cyclopent—l-ene—l~carbaldehyde obtained in Example 721 (79.9 mg, 0.831 mmol).

FP11-0417 1H—NMR (400 MHz, CD30D) 5 ppm; 0.90 (3H, d, J=7 Hz), 1.58-1.75 (2H, m), .00 (4H, m), 2.15-2.27 (1H, m), 2.47 (1H, d, J=16 Hz), 2.32-2.42 (4H, m), 2.46-2.65 (3H, m), 2.62 (1H, d, J=10 Hz), 2.95 (1H, t, J=9 Hz), 2.99 (1H, d, J=l6 Hz), 3.03-3.16 (2H, m), 3.36- 3.43 (2H, m), 3.51 (1H, d, J=10 Hz), 3.73—3.82 (1H, m), 3.98 (1H, d, J=13 Hz), 4.03 (1H, d, J=13 Hz), 5.78—5.83 (1H, br), 7.25 (1H, t, 1:55 Hz), 7.43 (1H, t, J=8 Hz), 7.61 (2H, d, J=8 Hz). 6 (Example 14) 2—'[(3S,4R)—1—'{[2-Chloro—6~(difluoromethyl)phenyl]methyl}~3—{[1— (cyclohexylmethyl)piperidin—4—y1]carbamoyl}-4~methy1pyrrolidin—3-y1]acetic acid (Example 14a) 1-(Cyclohexylmethy1)piperidinamine The title nd was obtained by a method similar to the method described in /0222175 A1 and Examples 4a and 4b. 1H—NMR (400 MHZ, CDCl3) 8 ppm: 0.77—0.87 (2H, m), 1.11-1.49 (8H, m), 1.61~1.80 (5H, m), 1.97 (2H, t, J=12 Hz), 2.14 (2H, d, J=7 Hz), 2.50-2.64 (1H, m), 2.96 (2H, d, J=12 Hz).

(Example 14b) tert~Buty1 2—[(3S,4R)—1-benzyl—3—{[1—(cyclohexy1methyl)piperidin— 4—yl]carbamoyl}—4—methylpyrrolidin—3—yl]acetate The title compound (250 mg, yield: 81.4%) was obtained from (3S,4R)—1-benzyl- 3~[2-(tert-butoxy)—2—oxoethy1]—4-methy1pyrrolidine—3-carboxy1ic acid obtained in Example 1j (200 mg, 0.6 mmol) and 1—(cyc1ohexylmethyl)pipe1idine—4~amine obtained in Example 14a (141 mg, 0.72 mmol) by a method r to the method described in Example 1k 1H—NMR (400 MHz, CDC13) 5 ppm; 0.80—0.98 (2H, m), 0.92 (3H, d, J=7 Hz), 1.07—2.15 (17H, m), 1, 58 (9H, s), 1.95 (1H, d, J=16 Hz), 2.10 (1H, d, J=7 Hz), 2.36 (1H, d, J=10 Hz), 2.55—2.83 (4H, m), 3.08 (1H, d, J=16 Hz), 3.59 (1H, d, J=10 Hz), 3.63—3.77 (1H, m), 3.64 (1H, d, J=13 Hz), 3.69 (1H, d, J=13 Hz), 7.21—7.36 (5H, m), 8.56 (1H, d, J=8 Hz).

(Example 14c) tert—Butyl 2—[(3S,4R)—1~{[2—chloro—6— (difluommethyl)phenyl]methyl}{[1-(cyclohexylmethyl)piperidin~4—yl]carbamoyl} methylpynolidin—3—yl]acetate The title compound (198 mg, yield: 67.9%) was obtained from tert—butyl 2- [(3S,4R)-1—benzy1—3-{[1-(cyclohexylmethyl)pipe1idin—4-y1]carbamoyl}—4-methylpynolidin— 3-yl]acetate obtained in Example 14b (250 mg, 0.489 mmol) and 2—(bromomethyl)—1-chloro~ 3-(difluoromethyl)benzene obtained by a method similar to that of Example 10f (250 mg, 0.978 mmol) by a method similar to the method bed in e 11. 1H—NMR (400 MHZ, CDC13) 8 ppm; 0.78-0.97 (2H, m), 0.91 (3H, d, J=7 Hz), 1.08-2.13 (18H, m), 1.40 (9H, s), 2.00 (1H, d, J=17 Hz), 2.05 (1H, d, J=7 Hz), 2.54 (1H, d, J=10 Hz), FPll-04l7 2.61 (1H, dd, J=6, 10 Hz), .77 (2H, m), 2.85 (1H, t, J=10 Hz), 3.60 (1H, d, J=17 Hz), 3.58-3.72 (1H, m), 3.92 (1H, d, J=13 Hz), 3.96 (1H, d, J=13 Hz), 6.97 (1H, t, J=55 Hz), 7.36 (1H, t, J=8 Hz), 7.51-7.58 (2H, m), 7.62 (1H, d, J=8 Hz).

(Example 14d) 2-[(3S,4R)-1—{[2-Chloro(difluoromethy1)phenyl]methyl}-3—{[1- hexylmethyl)piperidin—4—yl]carbamoyl}—4—methylpyrrolidin—3-yl]acetic acid The title compound (140 mg, yield: 79.8%) was obtained from tert-butyl 2— [(3S,4R)—1~{[2-chloro—6-(difluoromethyl)phenyl]methyl}~3-{[1— (cyclohexyhnethyl)piperidin—4—y1]carbamoyl}-4—methylpyrrolidin—3—yl]acetate obtained in Example 140 (194 mg, 0.325 mmol) by a method similar to the method described in Example 1m. 1H—NMR (400 MHz, CD3OD) 6 ppm; 0.90 (3H, d, J=7 Hz), 0.93-1.06 (2H, m), 1.16—1.40 (4H, m), 1.62-1.83 (8H, m), 1.85-1.98 (2H, m), 2.17—2.28 (1H, m), 2.29 (1H, d, J=16 Hz), .50 (1H, m), 2.56-2.80 (4H, m), 2.90-2.99 (2H, m), 3.10-3.25 (2H, m), 3.49 (1H, d, J=10 Hz), 3.76-3.86 (1H, m), 3.97 (1H, d, J=13 Hz), 4.01 (1H, d, J=13 Hz), 7.28 (1H, t, J=55 Hz), 7.42 (1H, t, J=8 Hz), 7.56-7.63 (2H, m).

(Reference Example 1) 1,3-Dibenzyl (3RS,4SR)—3-[2—(tert—butoxy)-2—oxoethyl]—4- methylpyrrolidine-l,3-dicarboxylate (Reference Example 1a) (3RS,4SR)—3-[2-(tert-Butoxy)—2—oxoethyl] methylpyrrolidinecarboxylic acid 2O 44.7 g of (4—methoxyphenyl)methyl (3RS,4SR)~1~benzyl[2—(tert—but0xy)—2— oxoethyl]—4-methylpy1rolidine—3—carboxylate was obtained by a method similar to the method described in es 2e to 2g. A part of this nd (20 g, 44.1 mmol) was dissolved in methanol (316 ml), 10% Pd/C (3.93 g) was added, and the atmosphere was replaced with hydrogen gas. The mixture was stirred at room temperature overnight and then stirred with addition ofwarm water (36°C, 160 ml) for 30 minutes, and the precipitated solid was dissolved. Pd/C was filtered off, the e was then concentrated so that about to 40 ml of water remained, and methanol (80 ml) was added to the cloudy residue containing water, which was stirred for 30 minutes. The precipitated solid was filtered off (LotA). lH—NMR ofLotA is shown below.

Lot B was obtained by a similar method using (4-methoxyphenyl)methyl (3RS,4SR)benzyl[2-(tert-butoxy)-2—oxoethyl]—4-methy1pyrrolidine—3-carboxylate (21 g, 46.3 mmol). After confirming that 1H—NMR ofLot B is identical to 1H—NMR ofLot A, LotA and Lot B were combined and dried to give the title compound (9.91 g).

FP11—0417 1H—NMR (400 MHz, D20) 5 ppm; 0.97 (3H, d, J=6 Hz), 1.42 (9H, s), 2.12-2.24 (1H, m), 2.29 (1H, d, J=17 Hz), 2.93 (1H, d, J=17 Hz), 3.04 (1H, t, J=12 Hz), 3.18 (1H, d, J=12 Hz), 3.49 (1H, dd, J=8, 12 Hz), 4.03 (1H, d, J=12 Hz).

(Reference e 1b) (3RS,4SR)—1-[(Benzyloxy)carbonyl][2—(tert—butoxy)— thyl]—4-methylpyrrolidine—3—carboxylic acid A 2 N aqueous sodium hydroxide solution (20.2 ml) was added to a mixture of (3RS,4SR)—3—[2—(tert—butoxy)—2~oxoethyl]methylpyrrolidinecarboxylic acid obtained in Reference Example 1a (9.84 g, 40.5 mmol), acetone (39 ml) and water (49 ml) with stirring under ice-cooling (0 to 1°C), and the on mixture was dissolved by stirring for 45 minutes. Benzyl chloroformate (6.35 ml, 44.5 mmol) and a 2 N s sodium hydroxide solution (22.3 ml) were simultaneously added dropwise to the reaction mixture at an internal temperature of 35°C or lower with stirring under ice-cooling (O to 1°C) over 20 minutes.

The reaction liquid was stirred in an ice bath and gradually returned to room temperature.

This was stirred at room temperature overnight. A 1 N aqueous sodium hydroxide solution (10 ml) was added to the reaction liquid with stirring under ice—cooling (internal temperature: about 15°C), and the mixture was adjusted to pH 12. This reaction liquid was returned to room temperature and washed three times by adding ethyl ether. The aqueous layer was adjusted to pH 2 to 3 by tially adding a 2 N aqueous hydrochloric acid solution (20.2 ml) and a l N aqueous hydrochloric acid solution (13 ml) with stirring under ice-cooling (internal temperature: 5°C or lower). The aqueous layer was returned to room temperature and ted with ethyl acetate three times. The organic layer was washed with brine, and then dried over sodium sulfate, filtered and concentrated. The residue was ved in ethyl acetate, washed with water four times and then with brine, dried over sodium sulfate, filtered and trated to give the title compound (14.53 g, yield: 95.1%). 1H—NMR (400 MHz, CDC13) 8 ppm; 1.02 (3H, t, J=8 Hz), 1.42 (9H, s), 2.14-2.22 (1H, m), 2.27 (1H, d, J=17 Hz), 3.04 (1H, d, J=5, 17 Hz), 3.12-3.19 (1H, m), 3.35 (1H, dd, J=7, 12 Hz), 3.65—3.72 (1H, m), 4.29 (1H, dd, J=7, 12 Hz), 5.09-5.19 (2H, m), 7.26—7.38 (5H, m).

(Reference Example 10) 1,3—Dibenzyl SR)[2-(tert-butoxy)—2-oxoethyl]— 4—methylpyrrolidine-1,3—dicarboxylate Benzyl bromide (4.48 ml, 37.7 mrnol) was added to a e of (3RS,4SR) [(benzyloxy)carbonyl][2-(tert-butoxy)-2—oxoethyl]—4—met1’1ylpyrrolidinecarboxylic acid obtained in Reference Example lb (14.5 g, 38.5 mmol), potassium carbonate (10.6 g, 77 mmol) and N,N—dimethylformamide (50 ml) with ng under ice-cooling (internal FP11-0417 temperature: 3 to 7°C), and the mixture was stirred for one hour, ed to room temperature and stirred overnight. Water was added to the reaction , which was extracted with ethyl acetate three times. The organic layers were combined, tially washed with a saturated s ammonium chloride solution (five times), water (twice) and brine, and then dried over sodium sulfate, filtered and concentrated to give the title nd (17.67 g, yield: 98.2%). 1H—NMR (400 MHz, CDC13) 5 ppm; 0.84-0.89 (3H, m), 1.36—1.39 (9H, m), 2.09218 (1H, m), 2.24 (1H, dd, J=3, 17 Hz), 3.04-3.13 (2H, m), 3.35 (1H, dd, J=8, 12 Hz), 3.59—3.68 (1H, m), 4.32 (1H, t, J=12 Hz), 506-5.20 (4H, m), 7.29—7.36 (10H, m).

[0241] (Reference Example 2) (3 S,4R)—3-[2-(tert—Butoxy)—2—0xoethyl]-4~ methylpyrrolidinecarboxylic acid (Reference Example 2a) (-)-Dibenzoyl—L—tartrate of (4-methoxyphenyl)methyl (3R,4R)-1—benzyl—4-methylpyrrolidjne-3—carboxy1ate (4-Methoxyphenyl)methyl RS)-1~benzyl—4—methylpyrrolidine—3— carboxylate tely obtained by a method similar to the method described in Example 2f (1000 mg, 2.946 mmol) was dissolved in methyl isobutyl ketone (4 ml), and benzoyl-L— tartaric acid (1055 mg) was added and dissolved. Crystals obtained from (4- methoxyphenyl)methyl (3RS,4RS)—1—benzyl—4—methylpyrrolidine—3—carboxylate separately obtained by a method similar to that of Example 2f and (-)—dibenzoyl-L—tartaric acid were added to the resulting solution as seed crystals, and the ing precipitate was filtered offto give the title compound (757 mg, yield: 36.8%). l (3.02 mL) was added to 755 mg ofthe resulting solid, which was heated and dissolved, and then tert—butyl methyl ether (6.04 mL) was added. The resulting precipitate was filtered off to give the title compound (658 mg, yield: 87.2%) as crystals. (4-Methoxypheny1)methyl (3RS,4RS)—1-benzyl-4—methylpyrrolidine—3— carboxylate separately obtained by a method similar to the method described in Example 2f (150.0 g, 441 mmol) was dissolved in methyl isobutyl ketone (600 ml), and (-)—dibenzoyl-L- tartaric acid (157.0 g) was added and dissolved with stirring. (4-Methoxyphenyl)methyl (3R,4R)-l-benzyl—4-methylpyrrolidine-3carboxylate (—)—dibenzoyl—L-tartrate obtained by the method described in the previous paragraph was added to the resulting solution as seed crystals (7.5 mg), followed by stirring for 18 hours and 18 minutes. The precipitated solid was filtered OEand washed with methyl isobutyl ketone (150 ml). The resulting solid was dried under d pressure at 50°C to give the title compound (152.07 g, yield: 49.4%).

FP11—0417 Ethanol (600 ml) was added to 150.00 g of the resulting solid, which was heated to 80°C with stirring, and ution of the solid was ed, after which heating was stopped.

Fifiy—nine minutes after stopping heating, tert-butyl methyl ether (300 ml) was added over nine minutes; after further six minutes, seed crystals (5 mg) were added. After 12 minutes, tert—butyl methyl ether (900 ml) was added over two hours and 38 minutes, followed by stirring for fiirther 10 hours and 43 s. The precipitated solid was filtered OE and washed with a mixture of l and tert-butyl methyl ether (75 ml + 150 ml). The resulting solid was dried under reduced pressure at 50°C to give the title compound (106.40 g, yield: 70.9%). 1H—NMR (400 MHz, DMSO—d6) 6 ppm; 1.05 (3H, d, J=6 Hz), 2.32-2.45 (2H, m), 2.64-2.78 (1H, m), 2.92-3.12 (3H, m), 3.75 (3H, s), 3.80-3.94 (2H, m), 5.04 (2H, dd, J=12, 16 Hz), 5.77 (2H, s), 6.90-6.96 (2H, m), 7.26-7.38 (7H, m), 7.52-7.58 (4H, m), 7.66-7.72 (2H, m), 7.95- 8.05 (2H, m).

Analysis by HPLC; (Analysis conditions) Column: CHIRALCEL OJ-H (manufactured by Daicel al Industries, Ltd.) (0.46 cm diameter x 25 cm), eluent: hexane/emanol/diethylanfine = 850/150/1 (v/V/v), flow rate: 1 ml/min., detection: UV (226 mn) sis result) The resulting title compound was analyzed under the above analysis conditions, and a peak with a retention time of 8.62 minutes (enantiomeric excess: 98.0% cc) and a peak with a retention time of 10.9 s were observed.

(Reference Example 2b) (4-Methoxypheny1)methy1 )benzyl—4— methylpyrrolidine—3-carboxylate Ethyl acetate (900 ml) was added to (—)—dibenzoyl—L—tartrate of (4- methoxyphenyl)methyl (3R,4R)—1-benzyl-4—methylpy1rolidine—3~carboxylate obtained in Reference Example 2a (104.0 g), and a 1 N aqueous sodium hydroxide solution (600 ml) was added with stirring. The aqueous layer was discarded, and the c layer was washed with water twice (100 ml, 50 ml). The resulting organic layer was concentrated under reduced pressure at 50°C to give the title compound (49.8 g, yield: 98.5%). 1H—NMR (400 MHz, CDCl3) 6 ppm; 1.12 (3H, d, J=7 Hz), 2.19 (1H, dd, J=6, 9 Hz), 2.44- 2.62 (2H, m), 2.73—2.84 (2H, m), 2.85-2.92 (1H, m), 3.55 (1H, d, J=13 Hz), 3.63 (1H, d, J=l3 Hz), 3.81 (3H, s), 5.02-5.10 (2H, m), .90 (2H, m), 7.21-7.33 (7H, m).

(Reference Example 2c) (3S,4R)[2-(tert-Butoxy)-2—oxoethyl]-4— methylpyrrolidine—3-carboxy1ic acid FPll-0417 Tetrahydrofinan (200 ml) was added to (4-methoxyphenyl)methyl (3R,4R)—1- benzyl-4—methylpyrrolidine—3-carboxylate obtained in Reference Example 2b (29.50 g, 87 mmol), ed by azeotropic dehydration. Tetrahydrofinan (200 ml) was finther added, followed by azeotropic dehydration. Tetrahydrofuran (400 ml) was added to this product, which was cooled in a dry ice-ethanol bath, and a lithium diisopropylamide/n—hexane— tetrahydrofuran solution (129 ml, 1.11 M, 144 rnmol) was added over 27 s. After 30 minutes, a solution of tert—butyl bromoacetate (21.20 g, 144 mmol) in tetrahydrofuran (30 ml) was added over seven minutes. After 39 minutes, a 20% s ammonium chloride solution (440 ml) was added, and ethyl acetate (440 ml) was further added to perform extraction The resulting organic layer was washed with Water twice (60 ml, 60 ml) and concentrated under reduced pressure at 30°C. Methanol (150 ml) and palladium hydroxide (885 mg) were added to the resulting concentrate, followed by stirring under en pressure (0.35 lVH’a) for seven hours and 20 minutes. Water (200 ml) and tetrahydrofuran (100 ml) were added to the reaction liquid, followed by filtration, and the catalyst was sequentially washed with ol (50 ml) and water (50 ml x 2). The filter washings were concentrated under reduced re at 50°C, the resulting aqueous layer was washed with tert-butyl methyl ether (100 ml), and the aqueous layer after washing was concentrated under reduced pressure at 50°C. Methanol (150 ml) was added to the ing residue, and the mixture was ultrasonically treated and then filtered. The resulting solid was dried under ‘20 reduced pressure at 50°C to give the title compound (8.16 g, yield: 38.5%). 1H—NMR (400 MHZ, D20) 5 ppm; 1.00 (3H, d, J=7 Hz), 1.45 (9H, s), 2.16-2.28 (1H, m), 2.33 (1H, d, J=17 Hz), 2.97 (1H, (1, 1:17 Hz), 3.08 (1H, t, J=12 Hz), 3.22 (1H, d, J=12 Hz), 3.50—3.58 (1H, m), 4.07 (1H, (1, 1:12 Hz).

(Reference Example 3) (3S,4R)~3~[2—(tert—Butoxy)-2—oxoethyl]—4~ methylpyrrolidine—3-carboxylic acid (R)-(-)—1,1'—binaphthy1—2,2'—diyl hydrogenphosphate ethanol solvate SR)-3~[2-(tert-Butoxy)oxoethyl]—4—methylpyrrolidine—3-carboxylic acid obtained by a method similar to that of Reference Example 1a (500 mg), (R)—(-)-1,1'— binaphthyl—2,2'-diyl hydrogenphosphate (359 mg), ethanol (10.0 mL) and water (10.0 mL) were sequentially added to a 50 mL round bottom flask, ed by stirring at room temperature for about 22 hours. The precipitated solid was collected by filtration and washed with a 1:1 ethanol-water mixture (2 mL). The wet product was dried under reduced pressure at 40°C for about one hour to give the title nd (269 mg, yield: 20.5%).

FP11—0417 1H-NMR (400 MHZ, DMSO-d6) 5 ppm:0.88 (3H, d, J=7 Hz), 1.06 (3H, t, J=7 Hz), 1.38 (9H, s), 2.09-2.16 (1H, m), 2.44 (1H, d, J=18 Hz), 2.74 (1H, t, J=12 Hz), 2.87 (1H, d, J=18 Hz), 3.11 (1H, d, J=l2 Hz), 3.41-3.47 (2H, m), 3.81 (1H, d, J=12 Hz), 4.36 (1H, t, J=5 Hz), 7.21 (2H, d, J=9 Hz), 7.28-7.31 (2H, t, J=8 Hz), 7.38-7.45 (4H, m), 8.02 (4H, t, J=8 Hz). 8.85 mg of the above white solid was weighed out in a screw glass vessel. 0.2 mL ofMilliQ water and 0.8 mL of 99.5% ethanol were added thereto. Thereafter, this was equally divided into three portions in 1.5 mL LCMS vials, and the vials were loosely capped and left to stand at room temperature. After nine days, crystals were ed to be precipitated in the vials. An X—ray diffraction experiment was performed with R—AXIS RAPID II (Rigaku Corporation) using the ing single crystals (0.40 x 0.40 x 0.06 mm).

The llographic data and structural analysis results are shown in Table 1, and the atomic coordinate data are shown in Tables 2 to 4. The absolute structure of the title compound was specified from such results.

[Table 1] t = 95.637 (2)° Volume 3346.4 (2) A Z value, calculated y 4, 1.357 g/cm Absorption coefiicient 12.406 cm”1 Crystal size 0.40 x 0.40 x 0.06 mm Maximum measured 20 l36.5° Total number ofreflections/ 35631/11955 [R (Strength)=0.0631] number ofunique reflections Completeness 98.5% Structure solution Direct method (SlR92) Refinement Least-squares method for F Data/parameter 835 Goodness offit indicator 1.103 R factor (all data) 0.0739 0.0477 -0.00 (3) 0.43 and —0.40 e/A [Table 2] FP11~0417 -x z y Wm) 0.2859(3) 0.0336(1) 167(3) 0.1343 (I) m—0.0428(3) 0.0300(2) 0.0464 (3) 0.1033 (1) 0.1764(4) 0.1547(2) 702(9) 0.5024(3) 0.2371 (3) 0.2258(2) 5.75 (7) 0.1574(3) 0.4519(2) 014 0.4273 (2) (3) (2) 0.3547(3) 0.0085 (4) 0.3501(2) 0.1998(2) (3) 0.3086(1) ~0.5826(4) (2) 0.4322(1) 0.3597(7) 0.2939(4) 0.016012) 0.0052 (4) 0.4183 (2) 0.0909(4) 0.3854(2) 0.0875(5) 0.326312) 0.1735 (6) 0.2967 (3) 0.2649(6) 0.3240(4) 8.6 (2) 0.2729(5) 0.3811(3) 7.3 (2) 0.1871 (4) 0.4143(3) 5.2 (1) 0.1966(5) 0.4725(3) 0.0627(3) 0.1192(4) 0.5031(2) 500(9) 0.1083(4) (2) 14.33 (8) C12 0.0971(4) 0.3927(2) ) C13 0.1444(3) 0.1916(4) 0.3603(2) 0.1966(4) 0.3516(2) 0.2881(5) 0.3224(3) 0.3792(5) 0.2969(3) 6.9 (2) 0.1655(5) 0.3780(5) 0.3048(3) C18 0.2101(4) 0.2867(5) 0.3367(2) 0.0837(3) 0.2113(4) FP11-0417 0.0923 (2) 349(7) 0.0561(4) 0.0784(2) 029 0.0149(4) 0.0295(4) (2) 0.1038(4) 0.0199(2) 5.28 (9) [Table 3] 0. 4005 (4) 0.1378 (2) 3.83 (7) 0.3697 (3) 0.1190 (2) 3.36 (7) 0.4594(4) 0.1223 (2) l 365 (7) -O.1383 (3) | 0.4398(4) 0.1003(2) | 4.30 (8) 0.2124(4) } 0.5278 (5) 0.1034(2) 15.5 (1) (5) 0.6406(5) 0.1268(3) 0.0872(5) 0.6641(4) 0.1475(3) 0.0064(4) 0.5749(4) 0.1465(2) ( ) C39 0.0979(4) 0.5978(4) 0.1669(2) 0.5145(4) 0.1631(2) ~0.2809 (4) 0.0451(2) 0.2360 (3) 0.0731(2) 9.2979(4) 0.0362 (2) 0.2987 (6) 0.0228 (2) C45 0.4161(3) 0.0981(4) 0.0671(2) 0.4261(3) 0.2755(4) 0.1337(2) 0.5158(4) 0.2244(4) 0.1718(2) 0.2262(4) 0.2444(5) 0.0384(2) (4) 0.1967(7) 0.2712(2) 6.9 (2) 0.6778(5) 0.2758(8) 0.2683(4) 11.6 (3) . 0.5293 (6) (9) 0.3237(3) 0.6039(6) 0.0633 (8) 0.2658(3) (3) 0.1337(4) 0.4509(2) 417(8) 0.3066(3) 0.0503(3) 0.4574(2) 328(6) 0.2645 (3) 0.1071 (4) 0.3633(4) (5) 527(9) 0.3424(3) 0.0754(3) 3.97 (7) (3) 0.0656(4) 0.4074(2) 3.86 (7) 0.2682(4) 0.0315(4) 0.3529(2) 459(8) C60 0.1912(4) 0.0265(5) 0.5417(2) 6.3 (1) .5 (1) 8.1(2) 0.4183 0.3988 0.0833 9.66 0.0381 0.1706 0.2575 9.35 0. 3213 0.3030 H7 0.0762 0.3357 | 0.3988 0.2466 0.4886 6.92 -—-—m_ H14 0.0067 0.1351 0.3665 6.33 0.4395 0.2749 8.32 [Table 4] 90919 0.0732 0.0400 0.0115 1130 0.0826 0.0090 -—-— 05777 92223 0.0486 5.38 0.3723 0.0427 7.84 FP11-0417 0.2771 13.90 0.2950 13.90 0.2814 0.3457 14.10 -0.2417 0.3134 14.10 12.22 0.5411 -0.0226 0.2509 12.22 . 5.00 .00 4.93 , 6.32 6.32 H58A 0.1662 H58B 4.9 i‘s‘i‘s 7.56 7.56 - 7.56 0.1922 0.2570 9.71 0.1250 0.1995 9.71 0.0759 0.2548 9.71 -O.1136 0.2050 8.68 H63B 0 1821 -0.1466 0.2443 8.68 H63C 02981 0.1297 0.2700 8.68 01146 0.1581 0.2229 10.35 0.0701 0.0253 0.2250 10.35 0.1370 I10.35 The absolute structures of the compounds of Examples 1 to 14 are specified and named based on the ation obtained in Reference Example 3.

The structural formulas ofExample Compounds 1 to 8 are as follows. cal Formula 13] FP11—0417 Example 1 * Chiral, stereochemistry was not determined Example 7 Example 8 The ural formulas ofExample Compounds 9 to 14 are as follows.

[Chemical Formula 14] FP11-0417 2' INIIIZ C6 ””05 CHF2 Example 10 : :CHF2 Example 13 Example 14 (Test Example 1) Inhibition of cell ion in fractalldne—induced chemotaxis assay (1) Method The inhibitory efi‘ects of the example compounds on fiactalldne—induced cell migration were ed using CX3CR1—transfected B300 cells After equilibrating the Transwell plate (24-well clusters, pore size: 5 pm, manufactured by Corning Incorporated), a fiactalkine solution (0.3 nM, manufactured by R&D Systems, Inc.) was added to the lower wells. CX3CR1-expressing B300 cells which FP11—0417 were preincubated with the test compound (0.001, 0.003, 0.01 or 0.03 pM) for 30 minutes were placed in the upper layer wells, followed by tion under the condition of5% C02 for 3.5 hours at 37°C. The number of cells migrated to the lower wells was evaluated using CellTiter (manufactured by Promega Corporation).

The inhibitory rate ofthe test compound on fi‘actalldne-induced cell migration was calculated by the following formula, where [A] is the number of migrated cells in the presence ofboth fractalldne and the test compound, [B] is the number ofmigrated cells in the presence of fractalkine and in the absence of the test nd, and [C] is the number of migrated cells in the absence of both fractalkine and the test compound; the 50% inhibitory concentration (IC50) was calculated based on the inhibitory rate.

Inhibitory rate (%) = [1—{(A—C)/(B—C)}] x 100 (2) Results The results ofthis test example are shown in the following table.

[Table 5] Test compound C50 ( Exam1e 1 —13 Exam.le 2 —21 Examle3 _ Example 6 Exarnple 8 3 __16 Example 11 6 —1—Example14 16

[0251] (Test Example 2) Amelioration ofbody weight loss in T cell transfer—induced colitis model (1) Method By using colitis-induced SCID mice in which sitive, CD45RB-high cells isolated fiom BALB/c mice cytes were injected, the y of the example compounds was evaluated by the body weight changes. The experiment was performed over 31 days. On Day 1, CD4-positive, CD45RB—high cells isolated from the spleen ofBALB/c mice (5 x 105 cells/mouse) were intravenously administered to SCID mice. From Day 16 to 31, the e compound was orally administered to the SCID mice once a day, FP11-0417 followed by measuring body weights ofall animals on Day 19, 22, 24, 26, 29 and 31.

The efiicacy was evaluated by body weight changes on Day 19, 22, 24, 26, 29 or 31. The body weight change (%) was determined by the formula shown below, where [A] is the body weight on Day 16, and [B] is the body weight on each day of body weight measurement (Day 19, 22, 24, 26, 29 or 31). body weight change (%) = B/A x 100 (2) Results The results are shown in Figs. 1 to 4. The abscissa in the figures indicates the number ofdays d, where the day on which CD4-posifive, CD45RB-high cells isolated 1'0 fiom the spleen of BALB/c mice ‘(5 x 1‘05 mouse) were in1IaVenously administered to SCID mice is Day 0.

Claims (24)

1. A compound represented by formula (1) or phannaceutically acceptable wherein R ents a C16 alkyl group unsubstituted or having 1 to 3 substituents selected from Substituent Group A, a C34; cycloalkyl group unsubstituted or having 1 to 3 substituents selected from Substituent Group A, or a C34; cycloalkenyl group unsubstituted or having 1 to 3 substituents selected fiom Substituent Group A, X represents a Cm alkyl group, 10 Y and Z are the same or different from each other and each ents a halogen atom or a C16 alkyl group unsubstituted or having 1 to 3 substituents selected from Substituent Group n represents 0 or 1, Substituent GroupA consists ofhalogen atoms, and 15 tuent Group B ts ofhalogen atoms.

2. The nd or phannaceutically able salt thereof according to claim 1, wherein R is a fluorobutyl group, a pentyl group, a cyclohexyl group, a difluorocyclohexyl group, a cyclopentenyl group or a cyclohexenyl group.

3. The compound or pharmaceutically acceptable salt thereof according to 2O claim 1 or 2, whereinX is a methyl group.

4. The compound or pharmaceutically acceptable salt thereof according to any one ofclaims 1 to 3, nY is a chlorine atom.

5. The compound or phannaceutically acceptable salt thereof according to any one of claims 1 to 4, wherein Z is a chlorine atom, a methyl group, a difluoromethyl 25 group or a trifluoromethyl group.

6. The compound or phannaceutically able salt thereof according to any one ofclaims 1 to 5, wherein n is 1.

7. Acompound according to claim 1, selected from the group consisting of: 2-[(3S,4R)— 1 — {[2-chloro(t1ifluoromethy1)phenyl]methyl}—3- {[ l -(2-fluoropentyl)piperidin— 4-y1]carbamoy1} hy1pyrrolidin—3~y1]acetic acid, 2-[(3S,4R)— 1 -[(2,6—dichloropheny1)methy1]( { 1 -[(4,4-difluorocyclohexyl)methyl]piperidin- 4-yl} carbamoyl)~4-methy1pyrrolidin—3~y1]acetic acid, 2-[(3S,4R)— 1 — {[2-chloro(t1ifluoromethyl)pheny1]methyl} {[ 1 -(cyclohex—1 -en-1 - yhnethyl)piperidin—4—y1]carbamoyl}—4—methylpy1rolidin~3~yl]acetic acid, 2~[(3S,4R)-1 hloro-6—methy1pheny1)methyl] {[1 ohex~1—en—1— yl)piperidin—4—y1]carbamoyl}—4—methylpyrrolidin—3~y1]acetic acid, 2~[(3S,4R)—1 — {[2—chloro-6~(t1ifluoromethy1)phenyl]methy1}—3~ { [ l -(cyclopent— 1 -en—1 - 10 ylmethy1)piperidin—4-yl]carbamoy1}—4—methylpyrrolidin—3-y1]acetic acid, 2-[(3S,4R)- l —[(2-chloro~6—methylpheny1)methyl]—3— {[(l -cyclopent— 1 -en— 1 - ylmethyl)piperidin—4—y1]carbamoyl}—4~methy1py1rolidin—3~y1]acetic acid, 2-[(3S,4R)—3- {[(3S)- l -(cyclohex— 1 —en— 1 -yhnethy1)py1rolidin—3-y1]carbamoyl} — 1 -[(2,6- dichloropheny1)methyl]-4~methy1pyrrolidin—3~yl]acetic acid, 15 2—[(3S,4R)— 1 - {[2-chloro—6—(difluoromethy1)pheny1]methyl}—3-[( 1 ~hexy1pipen'din—4— yl)carbamoy1]—4—methylpy1rolidjny1]acetic acid, , 2-[(3S,4R)—3—{[(1 -cyclohex-l-eny1methy1)pipe1idin—4-y1]carbamoyl}[(2,6— dichloropheny1)methy1]—4—methylpyrrolidin—3-y1]acetic acid, 2—[(3S,4R)— 1 — {[2-chloro-6—(difluoromethyl)pheny1]methyl}~3— {[ 1 -(cyclohex— 1 —en 20 ylmethyl)pipeiidin—4-yl]carbamoyl}-4—methylpyirolidin—3~yl]acetic acid, 2-[(3S,4R)-1 - {[2-chloro—6-(difluoromethyl)pheny1]methyl}—3- {[1 opent-1 -en-l - ylmethyl)piperidin—4—yl]carbamoyl}~4~methy1pyrrolidin~3-yl]acetic acid and 2—[(3S,4R)-l — {[2—chloro(difluoromethyl)pheny1]methyl }-3— {[ 1 — (cyclohexylmethyl)piperidin-4—y1]carbamoyl}—4~methy1pyrrolidin—3—yl]acetic acid, 25 or a phannaceutically acceptable salt thereof.

8. A medicine comprising the compound or phannaceutically acceptable salt thereofaccording to any one ms 1 to 7 as an active ingredient.

9. A therapeutic agent for inflammatory bowel disease comprising the compound or pharmaceutically acceptable salt thereof according to any one of claims 1 to 7 3O as an active ingredient.

10. The therapeutic agent according to claim 9, n the inflammatory bowel disease is ulcerative colitis or s disease.

11. A fiactalkine—CX3CR1 pathway inhibitor comprising the compound or phannaceutically able salt thereof according to any one of claims 1 to 7 as an active ient.

12. A fiactalkine inhibitor comprising the compound or pharmaceutically acceptable salt thereofaccording to any one ofclaims 1 to 7 as an active ingredient

13. A CX3CR1 inhibitor comprising the compound or pharmaceutically acceptable salt thereofaccording to any one ms 1 to 7 as an active ingredient

14. A compound according to claim 1, comprising 2—[(3S,4R)—1—[(2,6— Dichlorophenyl)methyl]-3—(-({1—[(4,4-difluorocyclohexyl)methyl]piperidin—4— yl}carbamoyl)4—methylpyrrolidin—3—yl]acetic acid or a phannaceutically acceptable salt 10 thereof:

15. A compound according to claim 1, comprising 2—[(3S,4R)—l-{[2—Chloro- 6-(trifluorometliyl)phenyl]methyl}—3-{[1-(cycloheX—l~en-l-ylmethlyl)pipeiidin—4— yl]carbamoyl}—4—methylpyrrolidin—3-yl]acetic acid or a pharmaceutically acceptable salt 15 thereof:

16. A compound according to claim 1, comprising 2-[(3S,4R)[(2-Chloro- y1phenyl)methyl] { [ l ohexenyhnethyl)piperidin-4—yl]carbamoy1} methylpyrrolidin—3-yl]acetic acid or a pharrnaceutically acceptable salt thereof:

17. A compound according to claim 1, comprising 2—[(3S,4R)—1-{[2—Chloro— fluoromethy1)phenyl]methyl}-3— {[1 ~(cyclopent—1—en—l—ylmethy1)piperidin—4— y1]carbamoy1}—4—methylpyrrolidin—3-yl]acetic acid or a phannaceutically acceptable salt thereof: HSQ-w O CI HN-CN of b

18. A compound according to claim 1, comprising 2—[3S,4R)-l~[(2—Chloro— 6—me’rhylpheny1)methyl]—3- {[(1 -cyclopent— 1 -en— 1 -ylmethy1)piperidin4—yl]carbamoyl}—4— methylpyn'olidinyl]acetic acid or a phannaceutically acceptable salt thereof:

19. A nd according to claim 1, comprising 2-[(3S,4R){[(3S) (CycloheX—l-en—l-yhnethy1)pyrrolidin—3—y1]carbamoy1}—1—[(2,6-dichlorophenyl)methyl] pyrrolidiny1]acetic acid or a pharmaceutically acceptable salt thereof:

20. A compound according to claim 1, comprising 2-[(3S,4R)—3-{[(1- Cyclohex—1-en—l -y1methyl)piperidin—4—yl]carbamoyl}-1 —[(2,6—dichlorophenyl)methy1]—4— methylpyrrolidin—3—y1]acetic acid or a pharmaceutically acceptable salt thereof:

21. The use of the compound according to any one of claims 1 to 7 or 14 to 20, in the production ofa medicament for treating atory bowel e.

22. Use ing to claim 21, wherein the inflammatory bowel disease is 10 ulcerative colitis or Crohn’s disease.

23. A compound according to any one of claims 1 or 14 to 20, substantially as herein described with reference to any one ofthe accompanying examples thereof.

24. Use according to claim 21, substantially as herein described with reference to any one ofthe examples thereof.