RU2772276C2 - Phenyldifluoromethyl-substituted prolineamide compound - Google Patents

Abstract

FIELD: pharmaceutics.

SUBSTANCE: invention relates to a compound of the formula (I) or its pharmaceutically acceptable salt, where, in the formula, R¹ is lower alkyl or halogen lower alkyl, R² is halogen or halogen lower alkyl, L is a bond or -CH₂-, A is CH or N, R³ is H or lower alkyl, R⁴ and R⁵ are the same or different and are H or lower alkyl, R⁶ and R⁷ are the same or different and are H, lower alkyl or halogen. The invention also relates to a pharmaceutical composition for the prevention or treatment of an autoimmune disease by means of inhibiting cathepsin S, containing the effective amount of a compound or its pharmaceutically acceptable salt of the compound of the formula I and one or several auxiliary substances. Compounds of the formula I are intended for the prevention or treatment of an autoimmune disease by means of inhibiting cathepsin S, wherein the autoimmune disease is selected from SLE and lupus nephritis, allergies or organ, bone marrow or tissue transplant rejections.

EFFECT: phenyldifluoromethyl-substituted prolineamide compound is obtained, inhibiting cathepsin S.

24 cl, 24 tbl, 18 ex

Description

The field of technology to which the present invention relates

[0001]

The present invention relates to a phenyldifluoromethyl-substituted prolinamide compound which has an inhibitory effect on cathepsin S and is expected to be used as an active ingredient in a pharmaceutical composition, for example, a pharmaceutical composition for the prevention and/or treatment of an autoimmune disease, including systemic lupus erythematosus (SLE ) and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection.

Background of the Invention

[0002]

Cathepsin S is a lysosomal cysteine protease expressed primarily in antigen-presenting cells such as dendritic cells, macrophages and B cells and causes degradation of the invariant chain associated with major histocompatibility complex class II (MHC class II) molecules during manufacture. Class II MHC molecules bind to an autologous or non-autologous peptide incorporated extracellularly and induce the secretion of various cytokines, presenting the autologous peptide or non-autologous peptide to CD4-positive T cells. It has been confirmed that inhibition or deletion of cathepsin S inhibits the loading of the antigenic peptide into MHC class II molecules, and, in addition, the suppression of antigen presentation to CD4-positive T cells reduces the immune response against foreign antigens ("Immunity", 1999, vol. 10, No. 2, pp. 207-217). It is believed that in the case of an autoimmune disease such as SLE, the antigen presentation described above occurs against a pathogenic autologous peptide, and therefore it is considered that there is a high probability that a cathepsin S inhibitor will be useful for the treatment of an autoimmune disease. ("Journal of Clinical Investigation", 1998, Vol. 101, No. 11, p. 2351-2363).

[0003]

Accordingly, a cathepsin S inhibitor is expected to be promising as an agent for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, or an agent for the prevention and/or treatment of allergy or organ, bone marrow, or tissue transplant rejection.

[0004]

Patent Document 1 discloses that the compound of formula (A) exhibits an inhibitory effect on cathepsin S, and is useful for treating various metabolic diseases or immune diseases such as SLE.

[Chem. one]

(Refer to this publication for formula symbols.)

Patent Document 2 discloses that the compound of formula (B) exhibits an inhibitory effect on cathepsin S and is useful for treating various metabolic diseases or immune diseases such as SLE.

[Chem. 2]

(Refer to this publication for formula symbols.)

Non-Patent Document 1 discloses that the compound of formula (C) prevents the development of SLE and lupus nephritis.

[Chem. 3]

Patent Document 3 discloses that a compound of formula (D) exhibits a cathepsin S inhibitory effect and is useful for treating diabetes mellitus and the like.

[Chem. 4]

((Refer to the present publication for the symbols in the formula.) Patent Document 4 discloses that the compound of formula (E) has an inhibitory effect on cathepsin S and is useful for treating diabetes mellitus and the like.

[Chem. 5]

(Refer to this publication for formula symbols.)

Materials used in the examination of the application

patent document

[0005]

[Patent Document 1] WO 2010/121918

[Patent Document 2] WO 2012/059507

[Patent document 3] WO 2010/142650

[Patent document 4] WO 2017/144483

Not a patent document

[0006]

[Non-Patent Document 1] Annals of the Rheumatic Diseases), 2015, Vol. 74, p.452-463

Disclosure of the present invention

Problems to be Solved by the Invention

[0007]

A compound is provided that has an inhibitory effect on cathepsin S and is expected to be useful as an active ingredient in a pharmaceutical composition, for example, a pharmaceutical composition for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies, or organ transplant rejection. , bone marrow or tissue.

Troubleshooting Tools

[0008]

As a result of extensive research on a compound having an inhibitory effect on cathepsin S, the present inventors have found that a phenyldifluoromethyl-substituted compound of prolinamide has an inhibitory effect on cathepsin S, thus the present invention is complete.

That is, the present invention relates to a compound of formula (I) or a salt thereof, and relates to a pharmaceutical composition containing a compound of formula (I) or a salt thereof and one or more excipients.

[Chem. 6]

(In the formula

R ¹ is lower alkyl or halo lower alkyl,

R ² is halo or halo lower alkyl,

L is a bond or -CH ₂ -,

A is CH or N,

R ³ is H or lower alkyl,

R ⁴ and R ⁵ are the same or different and are H or lower alkyl, and

R ⁶ and R ⁷ are the same or different and are H, lower alkyl or halogen.)

Unless otherwise described, in the event that the symbols in the chemical formulas in the present description are also used in other chemical formulas, these same symbols have the same meaning.

[0009]

The present invention further relates to a pharmaceutical composition for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection, comprising a compound of formula (I) or a salt thereof. The pharmaceutical composition includes an agent for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies or transplant rejection of an organ, bone marrow or tissue, and it contains a compound of formula (I) or a salt thereof.

The present invention further relates to:

(1) a compound of formula (I), or a salt thereof, which is a cathepsin S inhibitor;

(2) a compound of formula (I) or a salt thereof for use as a cathepsin S inhibitor;

(3) a cathepsin S inhibitor containing a compound of formula (I) or a salt thereof;

(4) the use of a compound of formula (I) or a salt thereof for the manufacture of a pharmaceutical composition for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection;

(5) the use of a compound of formula (I) or a salt thereof for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection;

(6) a compound of formula (I), or a salt thereof, for use in the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection; and

(7) a method of preventing and/or treating an autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection, the method comprising administering to the subject an effective amount of a compound of formula (I) or a salt thereof.

"Subject" refers to humans or other animals in need of prevention or treatment of a disease, and in one embodiment, subject refers to humans in need of prevention or treatment of a disease.

Implementation of the present invention

[0010]

The compound of formula (I) or a salt thereof has a cathepsin S inhibitory effect and can be used as an agent for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection.

Embodiments of the present invention

[0011]

Hereinafter, the present invention will be described in detail.

[0012]

The term "lower alkyl" refers to straight or branched alkyl having 1 to 6 carbon atoms (hereinafter also referred to as C _1-6 ), such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl and n-hexyl, and refers to C _1-4 alkyl according to one embodiment, methyl or ethyl according to one embodiment, and methyl according to one embodiment.

[0013]

The term "halogen" means F, Cl, Br and I.

[0014]

The term "halogen lower alkyl" is C _1-6 alkyl substituted with one or more halogen atoms, and refers to C _1-6 alkyl substituted with 1 to 5 halogen atoms according to one embodiment, and refers to CF ₃ according to one embodiment.

[0015]

Some aspects of the present invention are shown below.

(1) A compound or salt thereof, in which formula (I) is represented by the following formula (Ia).

[Chem. 7]

(2) A compound or salt thereof, in which the formula (I) is represented by the following formula (Ib).

[Chem. eight]

(3) A compound or salt thereof, wherein R ¹ is lower alkyl or halo lower alkyl; a compound or salt thereof, in which R ¹ is halo lower alkyl; a compound or salt thereof in which R ¹ is lower alkyl; a compound or salt thereof, in which R ¹ is methyl or CF ₃ ; or a compound or salt thereof wherein R ¹ is methyl.

(4) A compound or salt thereof, wherein R ² is halogen or halo lower alkyl; a compound or salt thereof, in which R ² is halogen; a compound or salt thereof, in which R ² is halo lower alkyl; or a compound or salt thereof, in which R ² is CF ₃ .

(5) A compound or salt thereof, wherein A is CH or N; a compound or salt thereof, in which A is CH; or a compound or salt thereof wherein A is N.

(6) A compound or salt thereof, wherein L is a bond or —CH ₂ —; a compound or salt thereof, in which L is a bond; or a compound or salt thereof, in which L represents -CH ₂ -.

(7) A compound or salt thereof, wherein R ³ is H or lower alkyl; a compound or salt thereof, in which R ³ is H; a compound or salt thereof, in which R ³ is lower alkyl; a compound or salt thereof, in which R ³ is methyl or ethyl; or a compound or salt thereof, in which R ³ is methyl.

(8) A compound or salt thereof, wherein R ⁴ is H or lower alkyl; a compound or salt thereof, in which R ⁴ is lower alkyl; or a compound or salt thereof wherein R ⁴ is H.

(9) A compound or salt thereof, wherein R ⁵ is H or lower alkyl; a compound or salt thereof, in which R ⁵ is lower alkyl; or a compound or salt thereof wherein R ⁵ is H.

(10) A compound or salt thereof, wherein R ⁶ is H, lower alkyl, or halogen; a compound or salt thereof, in which R ⁶ is lower alkyl; a compound or salt thereof, in which R ⁶ is halogen; or a compound or salt thereof wherein R ⁶ is H.

(11) A compound or salt thereof, wherein R ⁷ is H, lower alkyl, or halogen; a compound or salt thereof, in which R ⁷ is lower alkyl; a compound or salt thereof, in which R ⁷ is halogen; or a compound or salt thereof wherein R ⁷ is H.

(12) A compound or salt thereof which is represented by two or more consistent combinations among the embodiments described in (1) to (11).

[0016]

Examples of the compound or salt thereof of the present invention represented by the combinations in the above embodiment (12) include the following embodiments.

(13) A compound of formula (I) or a salt thereof, wherein R ¹ is lower alkyl or halo lower alkyl, R ² is halo or halo lower alkyl, L is a bond or -CH ₂ -, A is CH or N, R ³ is H or lower alkyl, R ⁴ is H or lower alkyl, R ⁵ is H or lower alkyl, R ⁶ is H, and R ⁷ is H.

(14) A compound of formula (Ia) or a salt thereof, wherein R ¹ is lower alkyl or halo lower alkyl, R ² is halo or halo lower alkyl, L is a bond or -CH ₂ -, A is CH or N, R ³ is H or lower alkyl, R ⁴ is H or lower alkyl, R ⁵ is H or lower alkyl, R ⁶ is H, lower alkyl or halogen and R ⁷ is H, lower alkyl or halogen .

(15) A compound of formula (I) or a salt thereof, wherein R ¹ is lower alkyl or halo lower alkyl, R ² is halo or halo lower alkyl, L is -CH ₂ -, A is CH or N, R ³ is H or lower alkyl, R ⁴ is H or lower alkyl, R ⁵ is H or lower alkyl, R ⁶ is H, lower alkyl or halogen, and R ⁷ is H, lower alkyl or halogen.

(16) A compound of formula (I) or a salt thereof, wherein R ¹ is lower alkyl or halo lower alkyl, R ² is halo lower alkyl, L is -CH ₂ -, A is N, R ³ is lower alkyl, R ⁴ is H, R ⁵ is H, R ⁶ is H, and R ⁷ is H.

(17) A compound of formula (Ia) or a salt thereof, wherein R ¹ is lower alkyl or halo lower alkyl, R ² is halo lower alkyl, L is -CH ₂ -, A is N, R ³ is lower alkyl, R ⁴ is H, R ⁵ is H, R ⁶ is H, and R ⁷ is H.

(18) A compound of formula (Ib) or a salt thereof, wherein R ¹ is lower alkyl or halo lower alkyl, R ² is halo lower alkyl, L is -CH ₂ -, A is N, R ³ is lower alkyl, R ⁴ is H, R ⁵ is H, R ⁶ is H, and R ⁷ is H.

[0017]

Specific examples of compounds included in the present invention include compounds or salts thereof selected from the following group:

(4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl) carbonyl]-L-prolinamide,

(4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1-{[1-(trifluoromethyl )cyclopropyl]carbonyl}-L-prolinamide,

(4R)-N-(1-cyanocyclopropyl)-4-[{4-[(4-ethylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}(difluoro)methyl]-1-{[1- (trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide and

(4R)-N-(1-cyanocyclopropyl)-4-[{4-[(4-ethylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}(difluoro)methyl]-1-[(1- methylcyclopropyl)carbonyl]-L-prolinamide.

[0018]

Specific examples of compounds included in the present invention include a compound or salt thereof, which is a crystal containing (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl )methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide (hereinafter referred to as “compound A” in some cases) and succinic acid in a 1:2 molar ratio, and which is characterized by any of the following aspects.

(1) The compound has approximately 2θ(°) peaks of 2.7, 5.3, 9.8, 10.4, 13.5, 14.0, 15.1, 16.6, 17.4, and 24.4 powder x-ray diffraction.

(2) The compound characteristically exhibits approximately 2θ(°) peaks of 5.3, 9.8, 15.1, 16.6, and 24.4 by powder X-ray diffraction.

(3) The compound is characterized by an endothermic peak of approximately 134.3° C. by differential scanning calorimetry (DSC analysis).

A crystal containing compound A and succinic acid in a 1:2 molar ratio also includes a crystal of compound A disuccinate and a co-crystal of succinic acid monosuccinate of compound A.

[0019]

Tautomers and geometric isomers may exist in the compound of formula (I) depending on the types of the substituent. According to the present description, the compound of formula (I) can be described in only one isomer form, but the present invention also includes isomers other than it, and includes a form obtained by separation of isomers, or a mixture thereof.

In addition, the compound of formula (I) may contain an asymmetric center or an axis of chirality in some cases, and enantiomers (optical isomers) based on it may exist. A compound of formula (I), or a salt thereof, includes any isolated single enantiomer, such as the (R) form or (S) form, and a mixture thereof (including racemic mixture or non-racemic mixture). In one embodiment, the enantiomer is "stereochemically pure". The term "stereochemically pure" refers to the degree of purity by which those skilled in the art will recognize that an enantiomer is substantially stereochemically pure. According to another embodiment, the enantiomer is, for example, a compound with a stereochemical purity of 90% e. and. (enantiomeric excess) or more, 95% e. and. or more, 98% e. and. or more or 99% e. and. or more.

[0020]

The salt of the compound of formula (I) is a pharmaceutically acceptable salt of the compound of formula (I), and depending on the types of substituents, an acid addition salt may be formed in some cases. Specific examples thereof include an acid addition salt of an inorganic acid such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, nitric acid and phosphoric acid; and organic acid such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, mandelic acid, tartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid, citric acid , methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, aspartic acid, and glutamic acid, and the like. In addition, the salt of the compound of formula (I) also includes a co-crystal of the compound of formula (I) and an acid.

[0021]

The present invention further includes crystal polymorphisms, various hydrates and solvates of the compound of formula (I) and a salt thereof. The present invention further includes a compound of formula (I), or a salt thereof, which is pharmaceutically acceptable and labeled with one or more radioactive or non-radioactive isotopes. Examples of suitable isotopes used for isotopic labeling of the compound of the present invention include isotopes such as hydrogen ( ² H, ³ H, etc.), carbon ( ¹¹ C, ¹³ C, ¹⁴ C, etc.) , nitrogen ( ¹³ N, ¹⁵ N, etc.), oxygen ( ¹⁵ O, ¹⁷ O, ¹⁸ O, etc.), fluorine ( ¹⁸ F, etc.), chlorine ( ³⁶ Cl, etc.) etc.), iodine ( ¹²³ I, ¹²⁵ I, etc.), phosphorus ( ³² P, etc.) and sulfur ( ³⁵ S, etc.).

The isotopically labeled compound of the present invention can be used for research and the like. tissue distribution of drugs and/or substrates. For example, radioactive isotopes such as tritium ( ³ H) and carbon 14 ( ¹⁴ C) can be used for this purpose in terms of ease of labeling and ease of identification.

Substitution with heavier isotopes, eg, replacement of hydrogen with deuterium ( ² H), is advantageous in relation to processing with improved metabolic stability in some cases (eg, increased in vivo half-life, reduced dose required, decreased drug-drug interactions).

Displacement with positron-study isotopes ( ¹¹ C, ¹⁸ F, ¹⁵ O, ¹³ N, etc.) can be used in a positron emission tomography (PET) test to investigate substrate receptor loading.

The isotopically labeled compound of the present invention can generally be prepared by methods of the prior art known to those skilled in the art, or by the same preparation method as in the Examples or Preparation Examples using suitable reagents that are isotopically labeled instead of non-isotopically labeled reagents.

[0022]

(Obtaining method)

Protective Groups in Organic Synthesis (4th edition, 2006)» by Wuts (PGM. Wuts) and Greene (T.W. Greene). Защитная группа может быть соответствующим образом выбрана и использована согласно таким реакционным условиям. В таком способе требуемое соединение может быть получено введением защитной группы для проведения реакции, а затем удалением защитной группы при необходимости.The compound of formula (I) and its salt can be obtained using various known synthetic methods using their basic structure or characteristics based on the types of substituents. Depending on the types of functional groups for the manufacturing process, in some cases it is efficient to replace the functional group with an appropriate protecting group (a group that can be easily converted into a functional group) in advance from the starting material to the intermediate. Examples of such a protecting group include a protecting group and the like. and are described in

Protective Groups in Organic Synthesis (4th edition, 2006)" by Wuts (PGM. Wuts) and Greene (TW Greene). The protecting group may be appropriately selected and used according to such reaction conditions. In such a method, the desired compound can be obtained by introducing a protecting group to carry out the reaction, and then removing the protecting group if necessary.

A pharmaceutically acceptable prodrug is a compound containing a group that can be converted into an amino group, a hydroxyl group, a carboxyl group or the like by solvolysis or under physiological conditions. Examples of a group forming a prodrug include the group described in Prog. Med., 5, 2157-2161 (1985) and "Pharmaceutical Research and Development, Drug Design, Hirokawa Publishing Company" (Hirokawa-Shoten Ltd., 1990), Vol. 7, Molecular Design 163-198.

In addition, like a protecting group, a prodrug of a compound of formula (I) can be prepared by introducing a particular group in a step from the starting material into an intermediate, or further reacting with the resulting compound of formula (I). The reaction can be carried out using a method known to the person skilled in the art, such as total esterification, amidation and dehydration.

The following describes a specific method for obtaining the compounds of formula (I). Each method of obtaining can also be carried out with a link to a reference document attached to the explanation. The production method of the present invention is not limited to the examples shown below.

[0023]

As used herein, the following abbreviations may be used.

DMF = N,N-dimethylformamide, DMSO = dimethyl sulfoxide, EtOAc = ethyl acetate, MeOH = methanol, MeCN = acetonitrile, THF = tetrahydrofuran, TEA = triethylamine, DIPEA = N,N-diisopropylethylamine, NMM = N-methylmorpholine, XPhos = 2- dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl, RuPhos = 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl, XantPhos = 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, HATU = O-(7 -azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate, saline = saturated aqueous sodium chloride solution, MgSO ₄ = anhydrous magnesium sulfate.

[0024]

The following abbreviations may be used in structural formulas and groups as described herein.

Ac=acetyl, BOC=t-butoxycarbonyl, t-Bu=t-butyl, Me=methyl, Et=ethyl, _CF3 =trifluoromethyl, Ms=methanesulfonyl, Ts=p-toluenesulfonyl, Tf=trifluoromethanesulfonyl, Ph=phenyl.

[0025]

(Obtaining method 1)

[Chem. nine]

(In the formula, R ^B is -BF ₃ ^- Y ⁺ , -B(OR) ₃ , etc. Lv is a leaving group. Y is an alkali metal such as Na or K. R can be H or lower alkyl or two R's can form a lower alkylene together.)

The compound of formula (I) can be obtained by a coupling reaction between compound (1) and compound (2). Examples of the leaving group include halogen, TfO, and the like.

In this reaction, Compound (1) and Compound (2) were used in equivalent amounts or either of them in excess. Their mixture was mixed in a solvent inert during the reaction in the presence of a base and a palladium catalyst at room temperature until heated to reflux temperature, according to one embodiment, from room temperature to 150°C, typically within 0.1 hour to 5 days .

Examples of the solvent include, but are not particularly limited to, aromatic hydrocarbons such as toluene, ethers such as THF and 1,4-dioxane, halogenated hydrocarbons such as dichloromethane, alcohols, DMF, DMSO, EtOAc, MeCN, H ₂ O and solvent mixtures. As an example of the base, an inorganic base such as CS ₂ CO ₃ , K ₃ PO ₄ , K ₂ CO ₃ , Na ₂ CO ₃ and KOH is preferable. Examples of the palladium catalyst include a palladium catalyst introduced into the system by palladium acetate and a phosphine ligand such as XPhos and RuPhos, palladium tetrakis(triphenylphosphine), palladium dichlorobis(triphenylphosphine), palladium(II) 1,1'-bis(diphenylphosphino) ) dichloride and the like.

[Document]

Edited by A.d. Meijere and F. Diederich, "Metal-Catalyzed Cross-Coupling Reactions", 1st edition, ^VCH Publishers Inc., 1997

Edited by The Chemical Society of Japan, "5th ^Edition , Courses in Experimental Science (Vol. 14)", Maruzen, 2005

[0026]

(Obtaining method 2)

[Chem. ten]

(In the formula, a compound of formula (I-1) is a compound of formula (I) in which L is CH ₂ and A is N.)

The compound of formula (I-1) can be obtained by introducing a leaving group into compound (3) and then reacting with compound (4).

In this reaction, the compound obtained by reacting compound (3) with a halogenated sulfonyl compound such as MsCl or TsCl in a solvent inert during the reaction in the presence of a base and compound (4) were used in equivalent amounts or either of them in excess quantity. Their mixture is mixed in a solvent inert during the reaction in the presence of a base under ice-cooling to reflux, preferably at 0° C. to 120° C., typically for 0.1 hour to 5 days.

Examples of the solvent include, but are not specifically limited to, aromatic hydrocarbons such as toluene, ethers such as 1,4-dioxane, halogenated hydrocarbons such as dichloromethane, DMF, DMSO, EtOAc, MeCN, and solvent mixtures. Examples of the base include an organic base such as TEA, DIPEA and NMM, an inorganic base such as K ₂ CO ₃ , Na ₂ CO ₃ and KOH, and the like.

[0027]

(Obtaining method 3)

[Chem. eleven]

The compound of formula (I) can be obtained by an amidation reaction between compound (5) and compound (6). In this reaction, compound (5) and compound (6) were used in equivalent amounts or either of them in excess. Their mixture is mixed in a solvent inert during the reaction in the presence of a condenser while cooling to warm, preferably at -20°C to 60°C, usually within 0.1 hour to 5 days. Examples of the solvent include, but are not particularly limited to, aromatic hydrocarbons such as toluene, ethers such as THF and 1,4-dioxane, halogenated hydrocarbons such as dichloromethane, alcohols, DMF, DMSO, EtOAc, MeCN, and solvent mixtures. . Examples of the capacitor include HATU, 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide or its hydrochloride, dicyclohexylcarbodiimide, 1,1'-carbonyldiimidazole, diphenylphosphoryl azide, and the like. In some cases, it is preferable to use an additive (eg 1-hydroxybenzotriazole) for the reaction. In some cases, it is preferable to carry out the reaction in the presence of an organic base such as TEA, DIPEA and NMM or an inorganic base such as K ₂ CO ₃ , Na ₂ CO ₃ and KOH and the like to allow the reaction to proceed smoothly.

Moreover, a method in which the compound (5) is converted into a reactive derivative and then reacted with the compound (6) can be used. Examples of the reactive carboxylic acid derivative include an acid halide obtained by reacting with a halogenating agent such as phosphorus oxychloride and thionyl chloride, a mixed acid anhydride obtained by reacting with isobutyl chloroformate or the like, an active ester obtained by condensing with 1 -hydroxybenzotriazole or the like, and the like. The reaction between such reactive derivatives and the compound (6) was carried out in a solvent inert during the reaction, such as halogenated hydrocarbons, aromatic hydrocarbons, ethers or the like. under conditions from cooling to heating, preferably at from -20°C to 60°C.

[Document]

S.R. Sandler and W. Karo, Organic Functional Group Preparations, 2nd ^Edition , Vol. 1, Academic Press, Inc., 1991

Edited by The Chemical Society of Japan, "Courses in Experimental Chemistry (5th ^Edition )", Vol. 16 (2005)

[0028]

(Synthesis of starting material 1)

[Chem. 12]

(In the formula, Lv is a leaving group, P _g1 and P _g2 are protecting groups, and X is halogen. The crossed double bonds in compound (13) are a mixture of geometric isomers.)

Examples of P _g1 include a BOC group and the like, and examples of P _g2 include a t-Bu group, an Et group, a Me group, and the like.

The steps represented by Step 1-1 to Step 1-3 are a reaction to obtain a compound (15) used as a Wittig reagent (phosphorus ylide) from a compound (17) and a reaction to obtain a compound (13) by a Wittig reaction of a compound (15 ) and compound (14), respectively. In Step 1-1 and Step 1-2, the compound (17) was allowed to react with a halogenating agent such as N-bromosuccinimide or bromine to form halide (16), and then the mixture with triphenylphosphine was stirred in a solvent inert during the reaction, at from cooling to heating under reflux, according to one variant of implementation at from 0°C to 120°C, usually within from about 0.1 hour to 5 days. In the case of dihalogenation during the halogenation reaction, the monohalide (16) can be obtained by reacting with diethylphosphonate. In Step 1-3, a mixture of compound (14) and compound (15) is stirred in a solvent inert during the reaction in the presence of a base at cooling to reflux, preferably at 0°C to 100°C, usually for from about 0.1 hour to 10 days. Examples of the solvent include aromatic hydrocarbons, ethers, halogenated hydrocarbons such as dichloromethane, alcohols, DMF, DMSO, EtOAc, MeCN, and mixtures thereof. Examples of the base include organic bases such as sodium methoxide, potassium t-butoxide, n-butyllithium, lithium hexamethyldisilazide, an inorganic base such as K ₂ CO ₃ , Na ₂ CO ₃ and KOH, and the like.

[Document]

Wittig, G et al., U. Ber., 1954, Vol. 87, p. 1318

The step represented by Step 1-4 is a reaction to obtain compound (12) by an oxidation reaction that occurs after the alkene hydroboration of compound (13). In this reaction, a reagent which is prepared by mixing a mixture of compound (13) and a borane-THF complex, 9-borabicyclo[3.3.1]nonane, disamylborane, texylborane or the like, in a solvent inert during the reaction, preferably at 10° C. to 80° C., typically over 0.1 hour to 3 days, treated with an equivalent amount or excess of an oxidizing agent in a solvent inert to the reaction, in the presence of a base at cooling to reflux, preferably at - 20°C to 80°C, usually within 0.1 hour to 3 days, and thus the compound (12) can be obtained.

Examples of the solvent include aromatic hydrocarbons, ethers such as THF, halogenated hydrocarbons, DMF, DMSO, EtOAc, MeCN, and mixtures thereof. Examples of the base include NaOH, K ₂ CO ₃ , Na ₂ CO ₃ and KOH, and the like. Examples of the oxidant include hydrogen peroxide, cumene hydroperoxide, peracetic acid, perbenzoic acid, meta-chloroperbenzoic acid, oxone, activated manganese dioxide, chromic acid, potassium permanganate, sodium periodate, and the like.

[Document]

J. Am. Chem., Soc., 1956, Vol. 78, p. 5694-5695

J. Org. Chem., 1986, Vol. 51, p. 439-445

The step represented by Step 1-5 is a reaction to obtain compound (11) by an oxidation reaction of compound (12). In this reaction, the compound (12) is treated with an equivalent amount or an excess amount of an oxidizing agent in a solvent inert during the reaction, under cooling to heating under reflux, preferably at -20°C to 80°C, usually within 0.1 hours to 3 days. This reaction suitably used DMSO oxidation such as tetrapropylammonium perruthenate (TPAP) oxidation or Swern oxidation or Dess-Martin oxidation. In the oxidation of TPAP, compound (12) was treated in the presence of tetrapropylammonium perruthenate, which was an oxidation catalyst, 4A molecular sieve, which was a dehydrating agent, and N-methylmorpholine-M-oxide (NMMO), which was a reoxidizing agent. Examples of the solvent include aromatic hydrocarbons, ethers, halogenated hydrocarbons such as dichloromethane, DMF, DMSO, EtOAc, MeCN, and mixtures thereof. Examples of other oxidizing agents include hydrogen peroxide, cumene hydroperoxide, peracetic acid, perbenzoic acid, meta-chloroperbenzoic acid, oxone, activated manganese dioxide, chromic acid, potassium permanganate, sodium periodate, and the like.

[Document]

J. Chem. Soc, Chem. Commun., 1987, p. 1625-1627

Edited by The Chemical Society of Japan, "5th ^Edition , Courses in Experimental Science (Vol. 14)", Maruzen, 2005

Protective Groups in Organic Synthesis)), 4^th edition, 2006, и т.п.The step represented by Step 1-6 is a deprotection reaction P _g1 of compound (11) and then condensation with compound (10), and thereby obtaining compound (9). The step can be carried out in the same manner as in Production Method 3 after carrying out the deprotection reaction by referring to the method described in "

Protective Groups in Organic Synthesis)), 4th ^edition , 2006, etc.

The step represented by Step 1 to 7 is a reaction to obtain compound (8) by fluorination of compound (9). In this reaction, the compound (9) was stirred in a solvent inert during the reaction in the presence of a fluorinating agent from cooling to heating, preferably at -20°C to 120°C, usually for 0.1 hour to 10 days. Examples of the solvent include aromatic hydrocarbons, ethers, halogenated hydrocarbons such as dichloromethane, DMF, DMSO, EtOAc, MeCN, and mixtures thereof. Examples of the fluorinating agent include 4-t-butyl-2,6-dimethylphenylsulfur trifluoride, hydrogen fluoride, diethylaminosulfur trifluoride (DAST), sulfur tetrafluoride (SF ₄ ), bis(2-methoxyethyl)aminosulfur trifluoride and the like.

[Document]

J. Am. Chem. Soc, 2010, Vol. 132, p. 18199-18205

Protective Groups in Organic Synthesis)), 4^th edition, 2006, и т.п.The step represented by step 1-8 is a reaction to obtain compound (7) by deprotecting compound (8). This reaction can be mentioned, for example, in the method described in "

Protective Groups in Organic Synthesis)), 4th ^edition , 2006, etc.

The step represented by Step 1 to 9 is a reaction to obtain a compound (1) by condensing a compound (7) and a compound (6). The step can be carried out in the same manner as Production Method 3.

[0029]

(Synthesis of starting material 2)

[Chem. thirteen]

(In formula P, _g3 represents a protecting group.)

Examples of P _g3 include 2,4,6-trichlorophenyl.

The step represented by Step 2-1 is a reaction to obtain a compound (18) by reacting a compound (1) and a compound (19). The step can be carried out in the same manner as Production Method 1, except that the compound (19) was used instead of the compound (2).

As the solvent, toluene or benzotrifluoride is preferred. As the base, TEA or tributylamine is preferred. As the palladium catalyst, a palladium catalyst adjusted in situ with palladium acetate and a phosphine ligand such as XantPhos is preferred.

[Document]

Organic Letters, 2012, Vol. 14, no. 20, pp. 5370-5373.

The step represented by Step 2-2 is a reaction to obtain a compound (3) by a reduction reaction of a compound (18). In this reaction, the compound (18) is treated with an equivalent amount or an excess amount of a reducing agent in a solvent inert during the reaction, under cooling to heating, preferably at -20°C to 80°C, usually within 0.1 hour to 3 hours. Examples of the solvent include aromatic hydrocarbons, ethers, halogenated hydrocarbons, alcohols such as MeOH, DMF, DMSO, EtOAc, MeCN, and mixtures thereof. Examples of the reducing agent include hydride reducing agents such as sodium borohydride and lithium borohydride.

[Document]

M. Hudlicky, "Reductions in Organic Chemistry, 2nd ^Edition (ACS Monograph: 188)", ACS, 1996

RC Larock, Comprehensive Organic Transformations)), 2nd Edition, ^VCH Publishers, Inc., 1999

T.J. Donohoe "Oxidation and Reduction in Organic Synthesis (Oxford Chemistry Primers 6)", Oxford Science Publications, 2000

Edited by The Chemical Society of Japan, "Courses in Experimental Chemistry (5 ^th Edition)))", Vol. 14 (2005)

[0030]

(Synthesis of starting material 3)

[Chem. fourteen]

The step shown in Step 3-1 is a reaction to obtain a compound (20) by a coupling reaction of a compound (8) and a compound (2), and can be carried out in the same manner as in the production method 1.

The step represented by step 3-2 is a reaction to obtain compound (5) by deprotection of compound (20), and can be carried out in the same manner as step 1-8 in the synthesis of starting material 1.

[0031]

The compound of formula (I) was isolated as a free form compound, salt, hydrate, solvate or crystal polymorphism thereof and purified. The salt of the compound of formula (I) can also be obtained by subjecting the compound to a salt formation reaction of the general method.

Isolation and purification were carried out using conventional chemical operations such as extraction, fractional crystallization and various fractional chromatography.

Different isomers can be obtained by selecting the appropriate starting compound or can be separated using differences in physicochemical properties between the isomers. For example, the optical isomer may be obtained by a general method of optical resolution of the racemic form (for example, fractional crystallization resulting in a diastereomeric salt with an optically active base or acid, chromatography using a chiral column, or the like, etc.) or may be derived from the corresponding optically active parent compound.

[0032]

The pharmacological activity of the compound of formula (I) can be confirmed by the following studies or well-known improved studies.

[0033]

Study Example 1: In Vitro Measurement of Human Cathepsin S Inhibiting Activity

In a 96-well plate, 5 μl of human cathepsin S enzyme (R&D 1183-CY-010) was added to 20 ng/well. Then, using assay buffer (50 mM sodium acetate, 250 mM sodium chloride and 5 mM dithiothreitol (DTT, pH=4.5)), the test compound (10 mM DMSO solution) was diluted by series 10 -fold dilutions with 5 steps so that the final concentration is from 0.1 nM to 1 μM, or through a series of 3-fold dilutions with 7 steps so that the final concentration is from 0.01 nM to 10 nM, and 10 μl is added to the well (final concentration of DMSO was 0.1%) followed by the addition of 25 μl of VVR-AMC synthetic substrate (Peptide Institute 3211-V) so that the final concentration is 40 μM, and thus the enzymatic reaction was initiated. The fluorescence intensity (excitation wavelength: 380 nm, fluorescence wavelength: 460 nm) was measured at 37°C for 5-10 minutes from the start of the reaction using a spectrofluorophotometer (SPECTRAMAX GEMINI, Molecular Devices), and the reaction rate at linearity (5 minutes) were obtained for each concentration of test compound. The degree of inhibition at each concentration was determined by suppressing the reaction rate during the absence of enzyme addition without test compound addition and the reaction rate during enzyme addition without test compound addition by 100% inhibition and 0% inhibition, respectively, and therefore the IC ₅₀ value was calculated from using the method of non-linear regression of the sigmoid Emax. The results are shown in Table 1. In Table 1, Ex is the Example Compound No. and Dat1 is the IC ₅₀ (nM) value of human cathepsin S inhibitory activity.

[0034]

[0035]

Study Example 2: Evaluation of the Inhibitory Effect on MHC Class II Expression in Vitro Using Mouse Splenocytes (Cell Scoring System)

In antigen-presenting cells, inhibition of cathepsin S suppresses the expression of class II MHC molecules. As a result, suppression of antigen presentation to CD4-positive T cells causes a deterioration in the immune response to foreign antigens. With regard to increasing the expression of MHC class II in B cells, the inhibitory effect of the compound of formula (I) was studied.

Splenocytes harvested from male C57BL/6J mice (Charles River Laboratories Japan, Inc.) were seeded in a 96-well plate at 1×10 ⁵ cells/well. With RPMI 1640 medium (containing 10% fetal bovine serum (FCS), 5 x 10 ^-5 M 2-mercaptoethanol, 50 IU/ml penicillin and 50 μg/ml streptomycin), a 10 mM DMSO solution of test compound was diluted by series 5 -fold dilutions with 9 steps so that the final concentration is from 0.026 nM to 10 μm, or diluted by a series of 5-fold dilutions with 12 steps so that the final concentration is from 0.205 pM to 10 μm (final concentration of DMSO is 0.1%) , and added. At the same time, LPS (Sigma L4005) was added to the well so that the final concentration was 2 μg/ml, and the cells were cultured at 37°C at 5% CO ₂ for 48 hours. After culturing, cells were stained with biotin labeled YAe antibody (EBIOSCIENCE 13-5741-85) at 4° C. for 20 minutes and washed. Cells were then further stained with fluorochrome FITC labeled anti-mouse B220 antibody (BD BIOSCIENCES 553088) and fluorochrome PE labeled streptavidin (BD BIOSCIENCES 554061) for 20 minutes at 4°C. Therefore, the level of expression (fluorescence intensity YAe-biotin/streptavidin-PE) of class II MHC associated with Ea peptide in B220 positive B cells was measured using a flow cytometry system (Guava EasyCyte Plus System, Millipore). The degree of inhibition at each concentration was determined by suppressing the value during the absence of LPS stimulation without the addition of test compound and the value during LPS stimulation without the addition of test compound by 100% inhibition and 0% inhibition, respectively, and therefore the IC ₅₀ value was calculated using the method non-linear regression of sigmoid Emax. The results are shown in Table 2. In the table, Ex is the Example Compound No. and Dat2 is the IC ₅₀ value (nM).

[0036]

[0037]

Study Example 3: Evaluation of the Inhibition Effect of Class II MHC Expression Ex Vivo Using Mouse Peripheral Blood

The effect of inhibiting the expression of MHC class II molecules was evaluated in an ex vivo system.

The test compound was orally administered to male C57BL/6J mice (Charles River Laboratories Japan, Inc.) and the inhibitory effect on increasing expression of MHC class II in peripheral blood B cells after oral administration was examined. That is, 10 ml/kg of test compound dissolved in vehicle [30% propylene glycol solvent {propylene glycol: hydrogenated castor oil (HCO40): Tween 80=4:2:1}/HCl (2 equivalents based on test compound)/water] , was administered orally to male C57BL/6J mice (dose: 0.3 mg/kg, 4 subjects per group), and peripheral blood was obtained after 6 hours. To 90 μl of peripheral blood was added 10 μl of PBS or 10 μl (final concentration 100 μg/ml) LPS (Sigma L4005) and cultivation was carried out at 37°C at 5% CO ₂ for 15 hours. After culturing, cells were stained with fluorochrome FITC-labeled anti-mouse IA/IE antibody (BD BIOSCIENCES 553623) and fluorochrome PE-labeled anti-mouse B220 antibody (BD BIOSCIENCES 553090) for 30 minutes under cooling, followed by hemolysis and fixation for 11-12 minutes. at 37°C using buffer (BD BIOSCIENCES Phosflow Lyse/Fix Buffer 558049). After washing using a flow cytometry system (FACSCanto II, BD BIOSCIENCES), the expression level of MHC class II on the surface of B220-positive B cells was measured with the average fluorescence intensity of FITC (hereinafter referred to as MFI) as an indicator. The difference between the MFI of LPS stimulation and the MFI without stimulation was defined as ΔMFI, and therefore the degree of ΔMFI inhibition according to test compound administration was calculated by setting the ΔMFI of mice treated with 10 ml/kg of vehicle as 1.

[0038]

In this study, the compound of example 8 showed 41% inhibition (0.3 mg/kg), which inhibited the increase in MHC class II expression.

[0039]

Study Example 4: Evaluation of the Inhibitory Effect on the Occurrence of SLE-Like Disease in a Spontaneous Occurrence Model Using F1 NZB/W Mice

NZB/W F1 mice (Japan SLC, Inc.) were used as a model for SLE, a spontaneously developing human-like disease "JAMA", 1966, Vol. 195, p. 145; "Advances in Immunology", 1985, Vol. 37, p. 269-390; "Journal of Biomedicine and Biotechnology", 2011, Vol. 2011, 271694).

Female NZB/W F1 mice were divided into groups according to urinary protein values (creatinine corrected value), plasma anti-dsDNA (IgG) antibody value, level of MHC class II expression in peripheral blood B cells, and body weight at 19 weeks of age. At this time, individuals with urinary protein (adjusted creatinine) values of 3 or more were excluded. At 20 weeks of age, test compound was administered orally twice a day, followed by urine collection and blood collection over time. Changes in urinary protein value and anti-dsDNA antibody value were assessed over time.

In the above study, the plasma value of anti-dsDNA IgG antibody produced in connection with SLE-like disease was measured by ELISA method (ALPHA DIAGNOSTIC 5120). In geometric mean values of the antibody at 28 weeks of age and at 32 weeks of age for each individual, the geometric mean of the 10 subjects treated with vehicle was 216662 U/mL, while the geometric mean of the 10 subjects treated with 1 mg/mL kg twice a day of the compound of example 2 was 26827 U/ml, which is a low value characterized by a significant difference (P value=0.0009, Dunnett's multiple comparison). In addition, urinary protein values (corrected for creatinine) were measured at 40 weeks of age. The geometric mean for the 10 subjects treated with the vehicle was 13.0, while the geometric mean for the 10 subjects treated with 1 mg/kg twice daily of the compound of Example 2 was 0.7, which is a low value characterized by significant difference (P-value = 0.0005, Dunnett's multiple comparison). Based on this, it was confirmed that the compound of Example 2 has an action of suppressing the occurrence of SLE-like disease in female NZB/W F1 mice.

[0040]

Study Example 5: Evaluation of Therapeutic Effects on SLE-Like Disease in an Induced Poly(I:C) Emergence Model Using NZB/WF1 Mice

poly(I:C), which is a ligand for Toll-like receptor 3, is administered to NZB/W F1 mice (Japan SLC, Inc.), and hence the increase in proteinuria associated with SLE-like disease can be accelerated. Administration of the test compound is started from a state where proteinuria is induced by administration of poly(I:C) and then the therapeutic effect on SLE-like disease is evaluated.

200 μg of poly (I:C) (InvivoGen tlrl-picw-250) was administered to 22 week old NZB/W F1 mice three times a week for 4 weeks for a total of 12 times. In the following 2 weeks, individuals whose urinary protein value (corrected for creatinine) became essentially 2 to 50 are included in the study and assigned based on the urinary protein value. After grouping, the test compound was administered orally twice a day for 5 weeks, urine was collected over time, and therefore the change in urinary protein value over time was evaluated.

[0041]

Based on the above results, the compound of formula (I), or a salt thereof, is expected to be useful as an agent for the prevention and/or treatment of an autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection.

[0042]

A pharmaceutical composition containing one or more compounds of formula (I) or salts thereof as active ingredient can be prepared using an excipient generally used in the art, i.e. a pharmaceutical agent, a pharmaceutical carrier, and the like, according to commonly used methods.

The administration may be in any form of oral administration with tablets, pills, capsules, granules, powders, solutions and the like, injections such as intra-articular, intravenous and intramuscular injections, and parenteral administration via suppository, eye drops, eye ointment, transdermal solution, ointment, transdermal patch, transmucosal solution, transmucosal patch, inhaler, and the like.

[0043]

As a solid oral composition, tablets, powders, granules, and the like are used. In such solid compositions, one or more active ingredients are admixed with at least one inert excipient. The composition may contain inactive additives such as lubricants and disintegrants, stabilizers and solubilizers, in accordance with the general methods. Tablets or pills may be sugar-coated or film-coated with a gastro-soluble or enteric-soluble substance, if desired.

Liquid compositions for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, elixirs, and the like, and include commonly used inert diluents such as purified water or ethanol. The liquid composition may contain a solubilizing agent, a wetting agent, an adjuvant such as a suspending agent, a sweetening agent, a flavoring agent, a flavoring agent and a preservative, in addition to an inert diluent.

[0044]

Injections for parenteral administration contain sterile aqueous or non-aqueous solutions, suspensions or emulsions. Examples of the aqueous solvent include distilled water for injection or saline. Examples of non-aqueous solvents include alcohols such as ethanol. Such a composition may further comprise a tonic, a preservative, a wetting agent, an emulsifying agent, a dispersing agent, a stabilizing agent, or a solubilizing agent. They are sterilized, for example, by filtration through a bacteria-retaining filter and mixing of a sterilizing agent or by irradiation. They may also be used by preparing a sterile solid composition and dissolving or suspending the composition in sterile water or a sterile injectable solvent prior to use.

[0045]

Examples of external preparations include ointments, patches, creams, jellies, hot compresses, sprays, lotions, eye drops, eye ointments, and the like. Commonly used ointment base, lotion base, aqueous or non-aqueous solutions, suspensions, emulsions, etc. .P.

[0046]

Transmucosal agents such as inhalers and transnasal preparations are solid, liquid or semi-solid and can be prepared according to known methods of the prior art. For example, well-known adjuvants may be appropriately added, and in addition, pH adjusters, preservatives, surfactants, lubricants, stabilizers, thickeners, and the like. For insertion, a correct inhalation or insufflation device may be used. For example, using a known device such as a metered dose inhaler device or a nebulizer, the compound can be administered alone or as a formulated powder, or as a solution or suspension in combination with a pharmaceutically acceptable carrier. The dry powder inhaler and the like may be a single or multiple administration device, and a dry powder or a powder capsule may be used. Alternatively, the form of administration may be a suitable means of release, such as a pressurized aerosol can, using a suitable gas, such as chlorofluoroalkane or carbon dioxide.

[0047]

In the case of oral administration, the daily dose is about 0.001 to 100 mg/kg, preferably 0.1 to 30 mg/kg, and more preferably 0.1 to 10 mg/kg of body weight, as appropriate, and this dose is administered one times or divided into 2-4 doses. In the case of intravenous administration, a suitable daily dose is from about 0.0001 to 10 mg/kg of body weight, and this dose is administered from one to several doses per day. With regard to the transmucosal agent, from about 0.001 to 100 mg/kg of body weight is administered from one to several doses per day. The dose is determined according to individual cases, taking into account symptoms, age, sex, and the like.

[0048]

The pharmaceutical composition of the present invention contains from 0.01% to 100 wt. % and from 0.01 to 50 wt. % according to one embodiment of one or more compounds of formula (I) or their salt, which is the active ingredient, although its weight may vary depending on the route of administration, dosage form, site of administration, types of excipients and additives.

[0049]

The compound of formula (I) can be used in combination with various agents for the treatment or prevention of diseases in which the compound of formula (I) is believed to be effective. The combination can be administered simultaneously or separately in turn or at a desired time interval. The agent for simultaneous administration may be a mixing agent or may be formulated separately.

[Examples]

[0050]

Hereinafter, the production method of the compound of formula (I) will be explained in more detail based on examples. The present invention is not limited to the compounds described in the following examples. In addition, each preparation method for the starting compound is shown in the Preparation Examples. Moreover, the production method of the compound of formula (I) is not limited to the production methods of the specific examples shown below. The compound of formula (I) may be prepared by a combination of such preparation methods, or by methods that are obvious to those skilled in the art.

[0051]

The initial temperature of the DSC curve, obtained by measurement under the following conditions, is shown in the following tables as the melting point.

DSC measurement was carried out using an aluminum sample cuvette in the uncoated state of the sample cuvette under the measurement conditions of temperature range: room temperature to 300°C, temperature increase: 10°C/min, and nitrogen flow rate: 50 ml/min using DSC Q2000 (measured by TA Instruments.).

[0052]

, измеренный диапазон дифракционного угла: (2θ): 2,5-40°.Powder X-ray diffraction was performed using RINT-TTR II (measured by RIGAKU Corporation) under lamp conditions: C, lamp current: 300 mA, lamp voltage: 50 kV, sample width: 0.020°, scan speed: 4°/min, wavelength : 1.5418

, measured diffraction angle range: (2θ): 2.5-40°.

As for the X-ray powder diffraction sample, since the nature of its data, the crystal lattice space and all samples are important in identifying the crystal identity, the error interval of the diffraction angle (2θ(°)) of the powder X-ray diffraction is usually ±0.2°, but the diffraction angle and diffraction intensity may vary depending on the direction of crystal growth, grain size and measurement conditions, and thus the samples do not have to be accurately interpreted.

[0053]

The following abbreviations were sometimes used in the examples, production examples, and tables described later.

Note. received = Receipt example no., approx. = example number, syn. = preparation method (indicating that the preparation was carried out in the same manner as described in Example No. or Production Example No.), p. = structural formula, given. = physico-chemical data quantity, ESI+ = m/z ESI-MS value (representing [M+H] ⁺ unless specifically stated otherwise), ESI- = m/z ESI-MS value (representing [M-H] ^- , unless specifically stated otherwise), NMR1: δ (ppm) peak in ¹ H NMR in DMSO-d ₆ at room temperature, NMR2: δ (ppm) peak in ¹ H NMR in DMSO-d ₆ at 80°C, NMR 3 : δ (ppm) peak in ¹ H NMR in DMSO-d ₆ at 60°C, [α] _D ^23.5 : D line, specific rotation at 23.5°C, mp: melting point, 2θ: diffraction peak angle in powder x-ray diffraction.

The HCl in the structural formula is the hydrochloride and the number in front of the HCl is the molar ratio. For example, 2HCl means dihydrochloride. Similarly, SA is succinate and 2SA is disuccinate (including a co-crystal containing the compound and succinic acid in a 1:2 molar ratio).

The symbol gm noted in the Production Example No. and Example No. is a mixture of geometric isomers. Similarly, the symbol em represents a mixture of epimers at the 4-position of the pyrrolidine ring or a mixture of epimers at the 3-position of the pyrrolidine ring, and the symbol dm represents a mixture of diastereomers with different steric configurations at the α-position of the benzyl group and the 4-position of the pyrrolidine ring.

[0054]

For convenience, the mol/l concentration is represented as M. For example, 1 M sodium hydroxide aqueous solution means 1 mol/l sodium hydroxide aqueous solution.

[0055]

Receiving Example 1

N-bromosuccinimide (148.9 g) and 47% hydrobromic acid (5 ml) were added to a MeCN (700 ml) solution of 4-bromo-1-methyl-2-(trifluoromethyl)benzene (100 g) at room temperature under a gaseous atmosphere argon. 2,2'-azobis(isobutyronitrile) (3.43 g) was added to the mixture, stirred at 70° C. for 10 minutes, and then stirred at 100° C. overnight. The mixture was cooled to room temperature. The mixture was cooled on ice, a saturated aqueous sodium thiosulfate solution was added, and the mixture was stirred for 10 minutes. Water and EtOAc were added to the mixture. The organic layer was separated and washed with saturated aqueous sodium bicarbonate and brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. n-hexane was added to the residue and the precipitated solid was separated by filtration. The filtrate was concentrated under reduced pressure. DIPEA (79 ml) and diethylphosphonate (54 ml) were added to a THF (500 ml) solution of the residue under ice-cooling. The mixture was stirred at room temperature for 2 hours. The mixture was cooled on ice and water was added thereto. EtOAc was added to the mixture and the organic layer was separated and washed with hydrochloric acid (1M), water, saturated aqueous sodium bicarbonate and brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. Triphenylphosphine (115 g) was added to a toluene (700 ml) solution of the residue and the mixture was stirred overnight at 100°C. The precipitate was collected by filtration and washed with toluene to give [4-bromo-2-(trifluoromethyl)benzyl](triphenyl)phosphonium bromide (198.51 g) as a solid.

[0056]

Receiving example 2

Potassium tert-butoxide (27.5 g) was added to a dichloromethane (600 ml) solution of [4-bromo-2-(trifluoromethyl)benzyl](triphenyl)phosphonium bromide (150 g) under argon gas under ice-cooling, and the mixture was stirred under room temperature for 6 hours. The mixture was cooled on ice, and a dichloromethane (150 ml) solution of 1-tert-butyl ester of (2S)-4-oxopyrrolidine-1,2-dicarboxylic acid 2-methyl ester (50 g) was added thereto, and the mixture was stirred at room temperature. temperature for 4 days. Saturated ammonium chloride aqueous solution was added to the mixture and stirred for 15 minutes.

The organic layer was separated and the aqueous layer was extracted with chloroform. The combined organic layers were washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. EtOAc (40 ml) and n-hexane (200 ml) were added to the residue and the mixture was stirred at room temperature for 1 hour. The precipitate was separated by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give 1-tert-butyl ester of 2-methyl ester (2S)-4-[4-bromo-2-(trifluoromethyl)benzylidene]pyrrolidine- 1,2-dicarboxylic acid (mixture of geometric isomers) (51.74 g) as an oily substance.

[0057]

Receiving example 3

Borane-THF complex (1 M THF solution, 167 ml) was added to THF (100 ml) solution of 1-tert-butyl ester of 2-methyl ester (2S)-4-[4-bromo-2-(trifluoromethyl)benzylidene ]pyrrolidine-1,2-dicarboxylic acid (mixture of geometrical isomers) (25.9 g) under argon gas with ice cooling. The mixture was stirred at 30° C. for 30 minutes. The mixture was cooled using an ice water bath to which sodium chloride was added, and MeOH (27 ml) was added thereto. A mixture of an aqueous solution of NaOH (1 M, 112 ml) and a 30% aqueous solution of hydrogen peroxide (18 ml) was added to the mixture, and the mixture was stirred at room temperature for 1 hour. The mixture was cooled on ice and EtOAc and 14% aqueous sodium thiosulfate was added. The mixture was stirred at room temperature for 1 hour. The organic layer was separated and washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give 1-tert-butyl ester of 2-methyl ester (2S)-4-{[4-bromo-2-(trifluoromethyl)phenyl]( hydroxy)methyl}pyrrolidine-1,2-dicarboxylic acid (mixture of diastereomers with different steric configurations at the α-position of the benzyl group and the 4-position of the pyrrolidine ring) (20.96 g) as a solid.

[0058]

Receiving Example 4

4-methylmorpholine N-oxide (6.1 g) and 4A molecular sieve (8.7 g) were added to a dichloromethane (105 ml) solution of 1-tert-butyl ester of 2-methyl ester (2S)-4-{[ 4-bromo-2-(trifluoromethyl)phenyl](hydroxy)methyl}pyrrolidine-1,2-dicarboxylic acid (mixture of diastereomers containing various steric configurations at the α-position of the benzyl group and the 4-position of the pyrrolidine ring) (20.96 g ) under nitrogen gas under ice-cooling, and the mixture was stirred for 10 minutes. Tetrapropylammonium perruthenate (1.5 g) was added to the mixture under ice-cooling, and the mixture was stirred at room temperature for 1 hour. EtOAc was added to the mixture and concentrated under reduced pressure. The precipitate was filtered using silica gel and washed with EtOAc. A saturated aqueous sodium thiosulfate solution was added to the filtrate, and the mixture was stirred. The organic layer was separated and washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give 1-tert-butyl ester of 2-methyl ester (2S)-4-[4-bromo-2-(trifluoromethyl)benzoyl]pyrrolidine- 1,2-dicarboxylic acid (mixture of epimers at the 4-position of the pyrrolidine ring) (19.6 g) as an oily substance.

[0059]

Receiving Example 5

Hydrogen chloride (4 M 1,4-dioxane solution, 395 ml) was added to MeOH (395 ml) suspension of 1-tert-butyl ester of 2-methyl ester (2S)-4-[4-bromo-2-(trifluoromethyl )benzoyl]pyrrolidine-1,2-dicarboxylic acid (mixture of epimers at the 4-position of the pyrrolidine ring) (78.9 g) under ice-cooling, and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure. Toluene was added to the mixture, and the mixture was concentrated under reduced pressure. 1-Methylcyclopropanecarboxylic acid (20 g), HATU (76 g) and DIPEA (86 ml) were added to a dichloromethane (630 ml) solution of the precipitate under nitrogen gas under ice-cooling, and the mixture was stirred for 5 hours at room temperature. The mixture was cooled on ice, and a saturated aqueous sodium bicarbonate solution was added thereto. Chloroform was added to the mixture and the organic layer was separated and washed with hydrochloric acid (1M), saturated aqueous sodium bicarbonate and brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by amino silica gel column chromatography (eluent; n-hexane/EtOAc) and silica gel column chromatography (eluent; n-hexane/EtOAc) to give 4-[4-bromo-2-(trifluoromethyl)benzoyl] methyl ester -1-[(1-methylcyclopropyl)carbonyl]-L-proline (mixture of epimers at the 4-position of the pyrrolidine ring) (57.65 g) as an oily substance.

[0060]

Receiving example 6

Using a reaction vessel made from Teflon (registered trademark), 4-tert-butyl-2,6-dimethylphenylsulfur trifluoride (138.6 g) and hydrogen fluoride-pyridine complex (101.5 ml) were added to dichloromethane (320 ml ) a solution of 4-[4-bromo-2-(trifluoromethyl)benzoyl]-1-[(1-methylcyclopropyl)carbonyl]-L-proline methyl ester (mixture of epimers at the 4-position of the pyrrolidine ring) (64 g) under atmosphere gaseous argon and the mixture was stirred for 15 hours at room temperature. Stirring in the reaction vessel was stopped and the mixture was allowed to stand at room temperature for 75 hours. The mixture was added to a mixture of ice and 28% aqueous ammonia solution (1400 ml). Chloroform was added to the mixture, and the mixture was stirred for 8 hours. The organic layer was separated and the aqueous layer was extracted with chloroform. The combined organic layers were washed with saturated aqueous sodium bicarbonate, hydrochloric acid (1M), saturated aqueous sodium bicarbonate and brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. n-hexane was added to the residue, and the mixture was stirred at room temperature for 1 hour. The insoluble matter was separated by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc). The resulting oily substance was mixed with n-hexane (253 ml) and EtOAc (2.5 ml). The mixture was heated and when a solid precipitated, the mixture was left to cool and stirred for 15 hours at room temperature. The precipitate was collected by filtration and washed with a mixture of n-hexane and EtOAc (99:1) to give the methyl ester (4R)-4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-[( 1-methylcyclopropyl)carbonyl]-L-proline (26.6 g) as a solid.

[0061]

Receiving Example 7

An aqueous solution of lithium hydroxide hydrate (1 M, 82 ml) was added to a mixture of methyl ester (4R)-4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-[(1-methylcyclopropyl )carbonyl]-L-proline (26.57 g) and THF (265 ml) under ice-cooling, and the mixture was stirred at room temperature for 2 hours. The mixture was cooled on ice and hydrochloric acid (1M, approx. 85 ml) was added thereto. EtOAc was added to the mixture and the organic layer was separated and washed with brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give (4R)-4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-[(1-methylcyclopropyl)carbonyl]- L-proline (26.03 g) as a solid.

[0062]

Receiving Example 8

1-Aminocyclopropanecarbonitrile hydrochloride (7.19 g), HATU (21.9 g) and DIPEA (23.5 ml) were added to a dichloromethane (258 ml) solution of (4R)-4-{[4-bromo-2-(trifluoromethyl )phenyl](difluoro)methyl}-1-[(1-methylcyclopropyl)carbonyl]-L-proline (25.80 g) under ice-cooling. The mixture was stirred at room temperature for 15 hours. Chloroform and a saturated aqueous ammonium chloride solution were added to the mixture. The organic layer was separated and washed with saturated aqueous sodium bicarbonate and brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give (4R)-4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-N-(1-cyanocyclopropyl) -1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide (28.68 g) as a solid.

[0063]

Receiving example 9

Using a reaction vessel made from Teflon (registered trademark), 4-tert-butyl-2,6-dimethylphenylsulfur trifluoride (25 g) and hydrogen fluoride-pyridine complex (18 ml) were added to a dichloromethane (64 ml) solution of methyl complex 4-[4-bromo-2-(trifluoromethyl)benzoyl]-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-proline ester (mixture of epimers at the 4-position of the pyrrolidine ring) (12.9 g) in an argon gas atmosphere and the mixture was stirred for 9 hours at room temperature. Stirring in the reaction vessel was stopped and the mixture was allowed to stand at room temperature for 14 hours. The mixture was stirred at room temperature for 11 hours. Stirring in the reaction vessel was stopped and the mixture was allowed to stand at room temperature for 14 hours. The mixture was added to a mixture of ice, 28% aqueous ammonia solution (220 ml) and chloroform and stirred for 3 hours. The organic layer was separated and the aqueous layer was extracted with chloroform. The combined organic layers were dried over anhydrous sodium sulfate and concentrated under reduced pressure. n-hexane was added to the residue, and the mixture was stirred at room temperature for 1 hour. The insoluble matter was separated by filtration, and the filtrate was concentrated under reduced pressure. Toluene was added to the residue and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give the methyl ester 4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-{[1-(trifluoromethyl )cyclopropyl]carbonyl}-L-proline (mixture of epimers at the 4-position of the pyrrolidine ring) (11.67 g) as an oily substance.

[0064]

Receiving example 10

4-{[4-Bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-proline methyl ester (mixture of epimers at the 4-position of the pyrrolidine ring ) (3.69 g) was purified by column chromatography (column: CHIRALPAK IA, eluent: n-hexane/EtOAc) to give (4R)-4-{[4-bromo-2-(trifluoromethyl)phenyl]( difluoro)methyl}-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-proline (1.91 g) as an oily substance.

[0065]

Receiving example 11

4-{[4-Bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-[(1-methylcyclopropyl)carbonyl]-L-proline 1,4-dioxane (10 ml) methyl ester mixture (mixture epimers at the 4-position of the pyrrolidine ring) (478 mg), potassium trifluoro[(4-methylpiperazin-1-yl)methyl]borate(1-) potassium (435 mg), palladium(II) acetate (23 mg), RuPhos (97 mg) and K ₃ PO ₄ (839 mg) and water (0.072 ml) were stirred overnight at 80° C. under argon gas. A saturated sodium bicarbonate aqueous solution was added to the mixture. Chloroform was added to the mixture and the organic layer was separated. The organic layer was washed with brine and concentrated under reduced pressure. The residue was purified by amino silica gel column chromatography (eluent; chloroform/MeOH) to give the methyl ester 4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1 -[(1-methylcyclopropyl)carbonyl]-L-proline (mixture of epimers at the 4-position of the pyrrolidine ring) (340 mg) as an oily substance.

[0066]

Receiving example 12

Triphenylphosphine (9.75 g) was added to a toluene (70 ml) solution of 4-bromo-1-(bromomethyl)-2-chlorobenzene (10 g) at room temperature, and the mixture was stirred at 80° C. for 6 hours. The mixture was cooled on ice and stirred for 30 minutes. The precipitate was collected by filtration and washed with toluene to give (4-bromo-2-chlorobenzyl)(triphenyl)phosphonium bromide (18.9 g) as a solid.

[0067]

Receiving example 13

2,4,6-Trichlorophenyl formate (1 g), palladium (II) acetate (120 mg), XantPhos (300 mg) and DIPEA (1.5 ml) were added to a toluene (30 ml) solution of 4-{[4-bromo -2-(trifluoromethyl)phenyl](difluoro)methyl}-N-(1-cyanocyclopropyl)-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (mixture of epimers at the 4-position of the pyrrolidine ring) ( 2.1 g) at 100° C. under argon gas and the mixture was stirred for 1 hour. The mixture was cooled on ice, and chloroform and brine were added thereto. The insoluble matter was separated by filtration with Celite, and the organic layer of the filtrate was separated. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane/EtOAc) to give 2,4,6-trichlorophenyl ester 4-{[(5S)-5-[(1-cyanocyclopropyl)carbamoyl]-1-{[ 1-(trifluoromethyl)cyclopropyl]carbonyl}pyrrolidin-3-yl](difluoro)methyl}-3-(trifluoromethyl)benzoic acid (mixture of epimers at the 3-position of the pyrrolidine ring) (2.4 g) as a solid.

[0068]

Receiving Example 14

Sodium borohydride (103 mg) was added to a mixture of 2,4,6-trichlorophenyl ester 4-{[(5S)-5-[(1-cyanocyclopropyl)carbamoyl]-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl} pyrrolidin-3-yl](difluoro)methyl}-3-(trifluoromethyl)benzoic acid (mixture of epimers at the 3-position of the pyrrolidine ring) (1 g) and THF (20 ml) with ice cooling. MeOH (2 ml) was added to the mixture under ice-cooling, and the mixture was stirred at room temperature for 30 minutes. Water was added to the mixture. EtOAc was added to the mixture and the organic layer was separated and washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give N-(1-cyanocyclopropyl)-4-{difluoro[4-(hydroxymethyl)-2-(trifluoromethyl)phenyl]methyl}-1-{ [1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (mixture of epimers at the 4-position of the pyrrolidine ring) (290 mg) as a solid.

[0069]

Receiving example 15

A mixture of 1,2-dibromoethane (0.032 ml) and chloro(trimethyl)silane (0.047 ml) was added to a suspension of zinc (365 mg) in N,N-dimethylacetamide (2 ml) at room temperature under argon gas, and the mixture was stirred at 50 °C for 15 minutes. An N,N-dimethylacetamide (2 ml) solution of 4-iodopiperidine-1-carboxylic acid tert-butyl ester (1.73 g) was added to the mixture at room temperature, and the mixture was stirred at 50° C. for 15 minutes. The above mixture was added to an N,N-dimethylacetamide (5 ml) suspension of 4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl methyl ester }-L-proline (a mixture of epimers at the 4-position of the pyrrolidine ring) (1 g), a complex of 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride-dichloromethane (151 mg) and copper (I) iodide ( 71 mg) at room temperature under argon gas and the mixture was stirred at 80° C. overnight. The mixture was cooled to room temperature, EtOAc was added and the insoluble matter was separated by filtration. The filtrate was washed with water and brine, and the organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: n-hexane/EtOAc) to give tert-butyl ester 4-[4-{difluoro[(5S)-5-(methoxycarbonyl)-1-{[1-(trifluoromethyl) cyclopropyl]carbonyl}pyrrolidin-3-yl]methyl}-3-(trifluoromethyl)phenyl]piperidine-1-carboxylic acid (mixture of epimers at the 3-position of the pyrrolidine ring) (1.15 g) as an oily substance.

[0070]

Receiving Example 16

Trifluoroacetic acid (1.5 ml) was added to a dichloromethane (15 ml) and MeCN (7.5 ml) solution of 4-[4-{[(5S)-5-[(1-cyanocyclopropyl)carbamoyl] tert-butyl ester -1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}pyrrolidin-3-yl](difluoro)methyl}-3-(trifluoromethyl)phenyl]piperidine-1-carboxylic acid (mixture of epimers at the 3-position of the pyrrolidine ring) ( 675 mg) under ice-cooling, and the mixture was stirred at room temperature for 3 hours. Trifluoroacetic acid (2.25 ml) was added to the mixture at room temperature, and the mixture was stirred for 4 hours. The mixture was added to a saturated aqueous sodium bicarbonate solution. Chloroform was added to the mixture and the organic layer was separated and washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure to give N-(1-cyanocyclopropyl)-4-{difluoro[4-(piperidin-4-yl)-2-(trifluoromethyl)phenyl]methyl}-1-{[ 1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (mixture of epimers at the 4-position of the pyrrolidine ring) (509 mg) as a solid.

[0071]

Receiving example 17

Potassium carbonate (5.5 g) was added to a dichloromethane (80 ml) solution of (4-bromo-2-chlorobenzyl)(triphenyl)phosphonium bromide (18.9 g) at room temperature under nitrogen gas, and the mixture was stirred for 20 minutes . (2S)-4-oxopyrrolidine-1,2-dicarboxylic acid 1-tert-butyl ester (2S)-4-oxopyrrolidine-1,2-dicarboxylic acid (6.5 g) and 18-crown-6 (110 mg) were added to the mixture at room temperature and the mixture stirred at reflux for 3 days. The mixture was cooled on ice, and a saturated aqueous sodium bicarbonate solution was added thereto. The organic layer was separated and washed with brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give 1-tert-butyl ester of 2-methyl ester (2S)-4-(4-bromo-2-chlorobenzylidene)pyrrolidine-1,2 -dicarboxylic acid (mixture of geometric isomers) (5.17 g) as an oily substance.

[0072]

Receiving Example 18

Bis(2-methoxyethyl)aminosulfur trifluoride (15 ml) was added to 4-(4-bromo-2-chlorobenzoyl)-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-proline methyl ester (mixture of epimers at 4-position of the pyrrolidine ring) (2.8 g) under an argon gas atmosphere, and the mixture was stirred at 90° C. for 3 hours. The mixture was cooled on ice and added to a mixture of ice and saturated aqueous sodium bicarbonate. EtOAc was added to the mixture and the organic layer was separated and washed with brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc) to give the methyl ester 4-[(4-bromo-2-chlorophenyl)(difluoro)methyl]-1-{[1-(trifluoromethyl)cyclopropyl] carbonyl}-L-proline (mixture of epimers at the 4-position of the pyrrolidine ring) (2.37 g) as an oily substance.

[0073]

Receiving example 19

Lithium hydroxide hydrate (20 mg) was added to MeOH (0.7 ml) and THF (0.7 ml) solution of 4-(4-{difluoro[(5S)-5-(methoxycarbonyl)-1-{ [1-(trifluoromethyl)cyclopropyl]carbonyl}pyrrolidin-3-yl]methyl}-3-fluorophenyl)piperidine-1-carboxylic acid (mixture of epimers at the 3-position of the pyrrolidine ring) (105 mg) at room temperature and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure to give (2S)-4-[{4-[1-(tert-butoxycarbonyl)piperidin-4-yl]-2-fluorophenyl}(difluoro)methyl]-1-{[1-(trifluoromethyl )cyclopropyl]carbonyl}lithium pyrrolidine-2-carboxylate (mixture of epimers at the 4-position of the pyrrolidine ring) (102 mg) as a solid.

[0074]

Example 1

Trifluoro[(4-methylpiperazin-1-yl)methyl]borate (1-)potassium (4.1g), XPhos (892mg), cesium carbonate (12g) and palladium(II) acetate (214mg) were added to 1,4-dioxane (50 ml) and aqueous (10 ml) solution of (4R)-4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-N-(1-cyanocyclopropyl)-1 -[(1-methylcyclopropyl)carbonyl]-L-prolinamide (5 g) at room temperature under argon gas, and the mixture was stirred for 3 hours under reflux. The mixture was cooled to room temperature and water was added thereto. The insoluble matter was separated by filtration with celite and washed with chloroform. Water and chloroform were added to the filtrate. The organic layer was separated and washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; n-hexane/EtOAc and chloroform/MeOH) to give (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl )methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide (4.7 g) as a solid.

[0075]

Example 2

Hydrogen chloride (4 M 1,4-dioxane solution, 1.39 ml) was added to EtOAc (7 ml) and ethanol (7 ml) solution of (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[ (4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide (1.43 g) and the mixture was stirred at room temperature for 30 minutes. The mixture was concentrated under reduced pressure. Ethanol was added to the residue and the mixture was concentrated under reduced pressure. EtOAc was added to the residue and the mixture was stirred for approximately 1 hour. The mixture was concentrated under reduced pressure. Ethanol (10 ml) was added to the precipitate and heated at 95°C to obtain a solution. The mixture was stirred overnight at room temperature. The precipitate was collected by filtration and washed with ethanol to give (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)- 1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide dihydrochloride (516 mg) as a solid.

[0076]

Example 4

Mixture of (4R)-4-{[4-bromo-2-(trifluoromethyl)phenyl](difluoro)methyl}-N-(1-cyanocyclopropyl)-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L- prolinamide (627.4 mg), potassium trifluoro[(4-methylpiperazin-1-yl)methyl]borate (1-) potassium (705 mg), palladium (II) acetate (25 mg), RuPhos (105 mg), K ₃ PO ₄ (905 mg), 1,4-dioxane (13 ml) and water (1.3 ml) were stirred at 80° C. for 6 hours under argon gas. Saturated sodium bicarbonate aqueous solution and chloroform were added to the mixture. The organic layer was separated and washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by amino silica gel column chromatography (eluent; chloroform/MeOH) to give (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2- (trifluoromethyl)phenyl}methyl)-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (491.4 mg) as an oily substance.

[0077]

Example 7

Hydrochloric acid (6 M, 0.678 mL) was added to an ethanolic (6 mL) solution of 4-{4-[{(3R,5S)-5-[(1-cyanocyclopropyl)carbamoyl]-1-[( 1-methylcyclopropyl)carbonyl]pyrrolidin-3-yl}(difluoro)methyl]-3-(trifluoromethyl)benzyl}piperazine-1-carboxylic acid (339 mg) and stirred at 50°C for 1 hour. The mixture was stirred at 60°C for 5 hours. The mixture was cooled on ice and an aqueous solution of NaOH (1 M, 5 ml) was added thereto. Chloroform was added to the mixture, and the organic layer was separated and concentrated under reduced pressure. The precipitate was purified by amino silica gel column chromatography (eluent; chloroform/MeOH). Hydrogen chloride (4 M 1,4-dioxane solution, 0.285 ml) was added to a solution of 1,4-dioxane (3 ml) and ethanol (5 ml) of the resulting oily substance, and the mixture was stirred at room temperature for 5 minutes. The mixture was concentrated under reduced pressure. EtOAc was added to the residue and the precipitate was collected by filtration and washed with EtOAc to give (4R)-N-(1-cyanocyclopropyl)-4-{difluoro[4-(piperazin-1-ylmethyl)-2-(trifluoromethyl)phenyl]methyl}- 1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide dihydrochloride (190 mg) as a solid.

[0078]

Example 8

A mixture of 4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-proline (mixture of epimers at 4 -position of the pyrrolidine ring) (415 mg), 1-aminocyclopropanecarbonitrile hydrochloride (117 mg), HATU (375 mg), DIPEA (0.338 ml) and DMF (8 ml) were stirred overnight at room temperature. Water was added to the mixture, and the mixture was extracted with chloroform. The organic layer was concentrated under reduced pressure. The precipitate was purified by amino silica gel column chromatography (eluent; chloroform/MeOH). The resulting oily substance was purified by reverse phase high performance liquid chromatography (eluent; 0.1% aqueous formic acid/MeCN). Hydrogen chloride (4 M 1,4-dioxane solution, 0.115 ml) was added to a dichloromethane (1 ml) solution of the resulting residue, and the mixture was stirred at room temperature for 5 minutes. The mixture was concentrated under reduced pressure to give N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1 -methylcyclopropyl)carbonyl]-L-prolinamide dihydrochloride (mixture of epimers at the 4-position of the pyrrolidine ring) (62 mg) as a solid.

[0079]

Example 9

Trifluoroacetic acid (0.11 ml) was added to dichloromethane (0.42 ml) and MeCN (0.11 ml) solution of 4-[4-{[(3R,5S)-5-[(l- cyanocyclopropyl)carbamoyl]-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}pyrrolidin-3-yl](difluoro)methyl}-3-(trifluoromethyl)benzyl]piperazine-1-carboxylic acid (35 mg) with ice-cooling and the mixture was stirred overnight at room temperature. The mixture was concentrated under reduced pressure. The precipitate was purified by amino silica gel column chromatography (eluent; chloroform/MeOH). Hydrogen chloride (4 M 1,4-dioxane solution, 0.015 ml) was added to a 1,4-dioxane (0.35 ml) solution of the resulting residue, and the mixture was stirred at room temperature for 5 minutes. The mixture was concentrated under reduced pressure, and diisopropyl ether was added to the residue. The precipitate was collected by filtration to give (4R)-N-(1-cyanocyclopropyl)-4-{difluoro[4-(piperazin-1-ylmethyl)-2-(trifluoromethyl)phenyl]methyl}-1-{[1-(trifluoromethyl )cyclopropyl]carbonyl}-L-prolinamide dihydrochloride (22 mg) as a solid.

[0080]

Example 10

Methanesulfonic chloride (0.011 ml) and TEA (0.039 ml) were added to a dichloromethane (2 ml) solution of N-(1-cyanocyclopropyl)-4-{difluoro[4-(hydroxymethyl)-2-(trifluoromethyl)phenyl]methyl}-1- {[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (a mixture of epimers at the 4-position of the pyrrolidine ring) (50 mg) and the mixture was stirred at room temperature for 30 minutes. The mixture was concentrated under reduced pressure. DMF (2 ml), 1-ethylpiperazine (0.024 ml) and potassium carbonate (86 mg) were added to the residue, and the mixture was stirred at room temperature for 20 hours. Water and EtOAc were added to the mixture and the organic layer was separated and washed with water and brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by amino silica gel column chromatography (eluent: n-hexane/chloroform and chloroform/MeOH) to give N-(1-cyanocyclopropyl)-4-[{4-[(4-ethylpiperazin-1-yl)methyl]-2 -(trifluoromethyl)phenyl}(difluoro)methyl]-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (mixture of epimers at the 4-position of the pyrrolidine ring) (52 mg) as a solid.

[0081]

Example 12

Hydrogen chloride (4 M EtOAc solution, 0.455 ml) was added to EtOAc (12 ml) and ethanol (1.2 ml) solution of (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazine -1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (565.9 mg) and the mixture was stirred at room temperature overnight. collected by filtration and washed with EtOAc to give (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1 -{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide dihydrochloride (508 mg) as a solid.

[0082]

Example 15

Acetic acid (0.010 ml) and sodium triacetoxyborohydride (50 mg) were added to a dichloroethane (1 ml) solution of N-(1-cyanocyclopropyl)-4-{difluoro[2-fluoro-4-(piperidin-4-yl)phenyl]methyl }-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (mixture of epimers at the 4-position of the pyrrolidine ring) (32 mg) under argon gas under ice-cooling, and the mixture was stirred for several minutes. Acetaldehyde (0.030 ml) was added to the mixture under ice-cooling, and the mixture was stirred for 30 minutes. Saturated aqueous sodium bicarbonate solution was added to the mixture under ice-cooling, and the organic layer was separated and washed with brine. The organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by amino silica gel column chromatography (eluent; n-hexane/EtOAc) to give N-(1-cyanocyclopropyl)-4-{[4-(1-ethylpiperidin-4-yl)-2-fluorophenyl](difluoro)methyl }-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (mixture of epimers at the 4-position of the pyrrolidine ring) (27 mg) as a solid.

[0083]

Example 17

Acetic acid (0.019 ml) and acetaldehyde (0.045 ml) were added to a dichloroethane (1.9 ml) solution of N-(1-cyanocyclopropyl)-4-{difluoro[4-(piperidin-4-yl)-2-(trifluoromethyl) phenyl]methyl}-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide (mixture of epimers at the 4-position of the pyrrolidine ring) (95 mg) at room temperature and the mixture was stirred for 5 minutes. Sodium triacetoxyborohydride (136 mg) was added to the mixture at room temperature, and the mixture was stirred for 2 hours. Water and chloroform were added to the mixture and the organic layer was separated and washed with brine. The organic layer was dried over MgSO ₄ and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent; chloroform/MeOH). Hydrogen chloride (4 M 1,4-dioxane solution, 0.025 ml) was added to a dichloromethane (2 ml) solution of the resulting oily substance, and the mixture was stirred for 30 minutes. The mixture was concentrated under reduced pressure to give N-(1-cyanocyclopropyl)-4-{[4-(1-ethylpiperidin-4-yl)-2-(trifluoromethyl)phenyl](difluoro)methyl}-1-{[1- (trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide hydrochloride (mixture of epimers at the 4-position of the pyrrolidine ring) (60.6 mg) as a solid.

[0084]

Example 18

Succinic acid (104 mg) was added to a 2-propanol (2.5 ml) solution of (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]- 2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide (250 mg) at room temperature. The mixture was stirred at 60° C. for 5 minutes and then stirred at room temperature overnight. 2-Propanol (2.5 ml) was added to the mixture and stirred for 10 minutes. The precipitate was collected by filtration and washed with a small amount of 2-propanol to obtain a crystal (281 mg) containing 1:2 (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl )methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide; and succinic acid (referred to herein as "disuccinate" in some instances).

[0085]

Compounds of preparation examples and examples shown in the following tables were prepared in the same manner as in the above preparation examples or examples.

[0086]

[0087]

[0088]

[0089]

[0090]

[0091]

[0092]

[0093]

[0094]

[0095]

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

Industrial Applicability

[0108]

The phenyldifluoromethyl-substituted prolinamide compound of the present invention has a cathepsin S inhibitory effect and is expected as an agent for the prevention and/or treatment of autoimmune disease, including SLE and lupus nephritis, allergies, or organ, bone marrow, or tissue transplant rejection.

Claims (44)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: [Chem. fifteen]

in formula R ¹ is lower alkyl or halo lower alkyl, R ² is halo or halo lower alkyl, L is a bond or -CH ₂ -, A is CH or N, R ³ is H or lower alkyl, R ⁴ and R ⁵ are the same or different from each other and represent H or lower alkyl, R ⁶ and R ⁷ are the same or different and are H, lower alkyl or halogen. 2. A compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein L is -CH ₂ -. 3. The compound or pharmaceutically acceptable salt thereof according to claim 2, wherein R ² is halo lower alkyl, A is N, R ³ is lower alkyl, R ⁴ is H, R ⁵ is H, R ⁶ is H and R ⁷ is H. 4. The compound or pharmaceutically acceptable salt thereof according to claim 1, where formula (I) is formula (Ia) [Chem. sixteen]

5. A compound or pharmaceutically acceptable salt thereof according to claim 1, wherein formula (I) is formula (Ib) [Chem. 17]

6. A compound or a pharmaceutically acceptable salt thereof according to claim 3, where formula (I) is formula (Ib). 7. A compound or pharmaceutically acceptable salt thereof according to claim 1, which is selected from the group consisting of: (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl) carbonyl]-L-prolinamide, (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}methyl)-1-{[1-(trifluoromethyl )cyclopropyl]carbonyl}-L-prolinamide, (4R)-N-(1-cyanocyclopropyl)-4-[{4-[(4-ethylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}(difluoro)methyl]-1-{[1- (trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide, (4R)-N-(1-cyanocyclopropyl)-4-[{4-[(4-ethylpiperazin-1-yl)methyl]-2-(trifluoromethyl)phenyl}(difluoro)methyl]-1-[(1- methylcyclopropyl)carbonyl]-L-prolinamide, and its pharmaceutically acceptable salt. 8. The compound or pharmaceutically acceptable salt thereof according to claim 7, which is (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2- (trifluoromethyl)phenyl}methyl)-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide or a pharmaceutically acceptable salt thereof. 9. The compound or pharmaceutically acceptable salt thereof according to claim 7, which is (4R)-N-(1-cyanocyclopropyl)-4-[{4-[(4-ethylpiperazin-1-yl)methyl]-2-( trifluoromethyl)phenyl}(difluoro)methyl]-1-{[1-(trifluoromethyl)cyclopropyl]carbonyl}-L-prolinamide or a pharmaceutically acceptable salt thereof. 10. The compound or pharmaceutically acceptable salt thereof according to claim 7, which is (4R)-N-(1-cyanocyclopropyl)-4-[{4-[(4-ethylpiperazin-1-yl)methyl]-2-( trifluoromethyl)phenyl}(difluoro)methyl]-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide or a pharmaceutically acceptable salt thereof. 11. The compound or pharmaceutically acceptable salt thereof according to claim 7, which is (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2- (trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide or a pharmaceutically acceptable salt thereof. 12. The compound or pharmaceutically acceptable salt thereof according to claim 11, which is (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2- (trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide disuccinate. 13. The compound or pharmaceutically acceptable salt thereof according to claim 12, which is a crystal of (4R)-N-(1-cyanocyclopropyl)-4-(difluoro{4-[(4-methylpiperazin-1-yl)methyl]-2 -(trifluoromethyl)phenyl}methyl)-1-[(1-methylcyclopropyl)carbonyl]-L-prolinamide disuccinate. 14. The compound or pharmaceutically acceptable salt thereof according to claim 13, which is a crystal characterized by peaks of approximately 2θ(°) 2.7, 5.3, 9.8, 10.4, 13.5, 14.0, 15 .1, 16.6, 17.4 and 24.4 by X-ray powder diffraction using a Cu lamp. 15. The compound or pharmaceutically acceptable salt thereof according to claim 14, which is a crystal characterized by an endothermic peak of 134.3°C differential scanning calorimetry. 16. Pharmaceutical composition for preventing or treating an autoimmune disease by inhibiting cathepsin S, containing an effective amount of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-15 and one or more excipients. 17. Pharmaceutical composition according to claim 16, wherein the autoimmune disease is selected from SLE and lupus nephritis, allergies or organ, bone marrow or tissue transplant rejections. 18. A pharmaceutical composition according to claim 16, wherein the compound or a pharmaceutically acceptable salt thereof is a compound or a pharmaceutically acceptable salt thereof according to claim 7. 19. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-15 for making a pharmaceutical composition for preventing or treating an autoimmune disease by inhibiting cathepsin S, the autoimmune disease being selected from SLE and lupus nephritis, allergy, or organ, bone marrow, or tissue transplant rejections. 20. Use according to claim 19, wherein the compound or pharmaceutically acceptable salt thereof is the compound or pharmaceutically acceptable salt thereof according to claim 7. 21. The use of a compound or a pharmaceutically acceptable salt thereof according to any one of paragraphs. 1-15 for preventing or treating an autoimmune disease by inhibiting cathepsin S, the autoimmune disease being selected from SLE and lupus nephritis, allergy, or organ, bone marrow, or tissue transplant rejections. 22. Use according to claim 21, wherein the compound or pharmaceutically acceptable salt thereof is the compound or pharmaceutically acceptable salt thereof according to claim 7. 23. A method of preventing or treating an autoimmune disease by inhibiting cathepsin S, wherein the autoimmune disease is selected from SLE and lupus nephritis, allergy, or organ, bone marrow, or tissue transplant rejections, the method comprising administering to a subject an effective amount of a compound or a salt thereof any of paragraphs. 1-15. 24. The method of claim 23, wherein the compound or pharmaceutically acceptable salt thereof is the compound or pharmaceutically acceptable salt of claim 7.