Arginase inhibitor and pharmaceutical composition comprising same

Abstract

The present invention relates to a compound represented by chemical formula 1 or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising same can be advantageously used for the prevention or treatment of cancer or tumors. [Chemical Formula 1] In chemical formula 1, R is as defined in the specification.

Description

Arginase inhibitor and pharmaceutical composition containing the same

[0001] The present invention relates to a compound of a novel structure that can be usefully used for the treatment or prevention of cancer or tumors.

[0002]

[0003] Regarding the development of immunotherapies, which are currently emerging in the field of cancer treatment, it is a well-known fact that arginine depletion in the tumor microenvironment is observed in cancer patients due to elevated levels of arginase. In fact, it is known that tumor cells, including those of acute myeloid leukemia, breast cancer, prostate cancer, glioblastoma, esophageal cancer, and renal cell carcinoma, evade destruction by the body's immune system through this mechanism. Consequently, there has been a need to develop arginase inhibitors that suppress arginase to restore arginine levels in the tumor microenvironment and promote the tumor-killing activity of cytotoxic T-cells.

[0004]

[0005] Regarding arginase inhibitors, pyrrolidine-based arginase inhibitors have been developed and are known to date, but there is a need for the development of arginase inhibitors with new chemical structures.

[0006]

[0007] Accordingly, the inventors of the present invention studied novel compounds having chemical structures different from arginase inhibitors reported to date, and confirmed that the compound described below exhibits excellent arginase inhibitory activity, thereby completing the present invention.

[0008]

[0009] The present invention is intended to provide a compound of a novel structure that can be usefully used for the treatment or prevention of cancer or tumors.

[0010]

[0011] To solve the above problem, the present invention provides a compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof:

[0012] [Chemical Formula 1]

[0013]

[0014] In the above chemical formula 1,

[0015] R is R1, -(CO)-CH(NH2)-R2, or -(CO)-R3, and

[0016] R1 is hydrogen, C 1-4 Alkyl, C 1-4 haloalkyl, amino, or -(C 1-4 It is an alkylene)amino, and

[0017] R2 is hydrogen, C 1-4 Alkyl, -(C 1-4 Alkylene)amino, phenyl, or benzyl, and

[0018] R3 is a 5-membered or 6-membered heterocycloalkyl containing at least one of N, O, and S.

[0019]

[0020] Preferably, R1 is hydrogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, 2,2,2-trifluoroethyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, or 4-aminobutyl. More preferably, R1 is hydrogen, methyl, ethyl, propyl, isopropyl, 2,2,2-trifluoroethyl, or 3-aminopropyl.

[0021]

[0022] Preferably, R2 is hydrogen, methyl, ethyl, propyl, isopropyl, 1-butyl, 2-butyl, isobutyl, tert-butyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, phenyl, or benzyl. More preferably, R2 is hydrogen, methyl, ethyl, propyl, isopropyl, 2-butyl, isobutyl, 4-aminobutyl, or benzyl.

[0023]

[0024] Preferably, R3 is pyrrolidinyl.

[0025]

[0026] Representative examples of compounds represented by the above chemical formula 1 are as follows:

[0027] 1) 1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0028] 2) 1-(4-voronobutyl)-5-methyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0029] 3) 1-(4-voronobutyl)-5-ethyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0030] 4) 1-(4-voronobutyl)-5-propyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0031] 5) 1-(4-voronobutyl)-5-isopropyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0032] 6) 5-(3-aminopropyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0033] 7) 5-(L-alanyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0034] 8) 5-(L-phenylalanyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0035] 9) 5-(L-lucyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0036] 10) 5-((2S)-2-amino-3-methylpentanoyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0037] 11) 1-(4-voronobutyl)-5-glycyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0038] 12) 5-(L-prolyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid,

[0039] 13) 5-(L-lysyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, and

[0040] 14) 1-(4-voronobutyl)-5-(2,2,2-trifluoroethyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid.

[0041]

[0042] Additionally, the compound of the present invention may exist in the form of a salt, particularly a pharmaceutically acceptable salt. As the salt, any salt commonly used in the art, such as an acid addition salt formed by a pharmaceutically acceptable free acid, may be used without limitation. The term "pharmaceutically acceptable salt" in the present invention means any organic or inorganic addition salt of said compound at a concentration having a relatively non-toxic and harmless active effect on the patient, such that the side effects caused by said salt do not impair the beneficial efficacy of the compound represented by Formula 1.

[0043]

[0044] Other pharmaceutically unacceptable salts or solvates of the compound represented by Formula 1 may be used as intermediates in the preparation of the compound represented by Formula 1, its pharmaceutically acceptable salts, or solvates.

[0045]

[0046] In addition, the compound represented by Formula 1 according to the present invention comprises, without limitation, its pharmaceutically acceptable salts as well as solvates such as possible hydrates that can be prepared therefrom and all possible stereoisomers. The solvates and stereoisomers of the compound represented by Formula 1 can be prepared from the compound represented by Formula 1 using methods known in the art.

[0047]

[0048] In addition, the compound represented by Formula 1 according to the present invention may be prepared in a crystalline or amorphous form, and if prepared in a crystalline form, it may be optionally hydrated or solvated. The present invention may include not only the stoichiometric hydrate of the compound represented by Formula 1 but also compounds containing varying amounts of water. The solvates of the compound represented by Formula 1 according to the present invention include both stoichiometric solvates and non-stoichiometric solvates.

[0049]

[0050] In addition, for example, the compound represented by Chemical Formula 1 according to the present invention can be prepared as shown in Reaction Scheme 1 below:

[0051] [Reaction Equation 1]

[0052]

[0053] The first reaction of the above reaction scheme 1 is an R1 substitution reaction, and the second reaction is a protecting group release reaction; the above manufacturing method can be further specified in the manufacturing examples to be described later.

[0054]

[0055] In addition, the compound represented by Formula 1 above, or a pharmaceutically acceptable salt thereof, has an arginase inhibitory effect and can restore arginine levels in the tumor microenvironment and promote the tumor-killing activity of cytotoxic T-cells by inhibiting arginase. Accordingly, the present invention provides a pharmaceutical composition for the prevention or treatment of cancer or tumor comprising the compound represented by Formula 1 above, or a pharmaceutically acceptable salt thereof.

[0056]

[0057] The term "prevention" in this invention refers to any act of suppressing or delaying the occurrence, spread, and recurrence of the said disease through the administration of the composition of this invention, and "treatment" refers to any act of improving or beneficially altering the symptoms of the said disease through the administration of the composition of this invention.

[0058]

[0059] The pharmaceutical composition of the present invention may be formulated into oral or parenteral administration forms in accordance with standard pharmaceutical practices. These formulations may contain, in addition to the active ingredient, pharmaceutically acceptable additives such as carriers, adjuvants, or diluents.

[0060]

[0061] Suitable carriers include, for example, physiological saline, polyethylene glycol, ethanol, vegetable oil, and isopropyl myristate, and diluents include, for example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and / or glycine, but are not limited thereto. In addition, the compounds of the present invention may be dissolved in oils, propylene glycol, or other solvents commonly used in the preparation of injectable solutions. Furthermore, the compounds of the present invention may be formulated into ointments or creams for topical action.

[0062]

[0063] The preferred dosage of the compound of the present invention varies depending on the patient's condition and body weight, the severity of the disease, the form of the drug, the route of administration, and the duration, but can be appropriately selected by those skilled in the art. However, for a desirable effect, it is preferable to administer the compound of the present invention at a dose of 0.0001 to 100 mg / kg (body weight) per day, preferably 0.001 to 100 mg / kg (body weight). Administration may be administered once a day or in divided doses via oral or parenteral routes.

[0064]

[0065] Depending on the method of administration, the pharmaceutical composition may contain 0.001 to 99 weight%, preferably 0.01 to 60 weight%, of the compound of the present invention.

[0066]

[0067] The pharmaceutical composition according to the present invention may be administered to mammals, including rats, mice, livestock, and humans, by various routes. All modes of administration are expected, for example, orally, rectally or intravenously, intramuscularly, subcutaneously, intrathecally, or intracervically.

[0068]

[0069] The compound represented by Formula 1 of the present invention, or a pharmaceutically acceptable salt thereof, may be usefully used for the prevention or treatment of cancer or tumors.

[0070]

[0071] Hereinafter, preferred embodiments are presented to aid in understanding the present invention; however, the following embodiments are merely illustrative of the invention and the scope of the invention is not limited to the following embodiments.

[0072]

[0073] Example 1: Preparation of rac-(3aS,6aR)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid

[0074]

[0075]

[0076] Step 1) Preparation of 5-((9H-fluorene-9-yl)methyl) 2-benzyl(1S,3aS,6aR)-1-(hydroxymethyl)tetrahydropyrrolo[3,4-c]pyrrole-2,5(1H,3H)-dicarboxylate

[0077] 5-((9H-fluorene-9-yl)methyl) 2-(tert-butyl) (1S,3aS,6aR)-1-(hydroxymethyl)tetrahydropyrrolo[3,4-c]pyrrole-2,5(1H,3H)-dicarboxylate was dissolved in DCM (133 mL) solvent, TFA (33 mL) was added, and the mixture was stirred at room temperature for 30 minutes. After confirming that the starting material disappeared on MS, the mixture was concentrated under reduced pressure. The resulting yellow oily compound was dissolved in DCM (117 mL), and NaHCO3 (12 g) and Cbz-OSu (6.4 g) were added and the mixture was stirred at room temperature. After confirming that the starting material disappeared on TLC, the mixture was extracted with a sat. NaHCO3 / DCM solvent. The extracted organic layer was dried over anhydrous MgSO4 and concentrated under reduced pressure, and column separated with ethyl acetate:hexane (1:9) to obtain a white solid compound 1-1 (6.9 g).

[0078] MS: [M+H] + = 499

[0079] 1 H NMR (500 MHz, CDCl3) δ 7.65 (s, 2H), 7.49 - 7.44 (m, 2H), 7.33 - 7.11 (m, 9H), 5.04 (s, 2H), 4.38 - 3.96 (m, 3H), 3.79 - 3.42 (m, 6H), 3.37 - 3.04 (m, 3H), 2.79 (s, 1H), 2.50 (s, 1H).

[0080]

[0081] Step 2) Preparation of (1S,3aS,6aR)-5-(((9H-fluorene-9-yl)methoxy)carbonyl)-2-((benzyloxy)carbonyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid

[0082] Compound 1-1 (6.9 g) was dissolved in acetone (70 mL) and the temperature was lowered to 0°C. 2.5 M Jones reagent (16.7 mL) was slowly added, and the mixture was stirred for 1 hour at the same temperature. After confirming that the starting material had disappeared on the MS, IPA was added and the mixture was stirred. The solution was diluted with ethyl acetate and filtered using Celite. The filtered solution was concentrated under reduced pressure and separated by column with methanol:dichloromethane (1:9) to obtain a pale yellow oil-like compound 1-2 (4.4 g).

[0083] MS: [M+H] + = 513

[0084]

[0085] Step 3) Preparation of 5-((9H-fluorene-9-yl)methyl) 1,2-dibenzyl(1S,3aS,6aR)-tetrahydropyrrolo[3,4-c]pyrrole-1,2,5(1H,3H)-tricarboxylate

[0086] Compound 1-2 (4.5 g) was dissolved in DMF (43 mL), potassium carbonate (1.4 g), sodium iodide (260 mg), and benzyl bromide (1.2 mL) were added, and the mixture was stirred at room temperature. After confirming that the starting material disappeared on MS, it was extracted with sat. NaHCO3 and ethyl acetate. The extracted organic layer was dried on anhydrous MgSO4, concentrated under reduced pressure, and separated by column with ethyl acetate:hexane (1:9) to obtain compound 1-3 (3.4 g) in a white solid state.

[0087] MS: [M+H] + = 603

[0088] 1H NMR (500 MHz, CDCl3) δ 7.74 (d,J= 8.0 Hz, 2H), 7.56 (d,J= 7.5 Hz, 2H), 7.41 - 7.21 (m, 14H), 5.33 - 4.97 (m, 4H), 4.49 - 4.14 (m, 4H), 3.91 - 3.18 (m, 6H), 2.92 (d,J= 35.6 Hz, 2H).

[0089]

[0090] Step 4) Preparation of dibenzyl(1S,3aS,6aR)-hexahydropyrrolo[3,4-c]pyrrole-1,2(1H)-dicarboxylate

[0091] Compound 1-3 (3.4 g) was dissolved in MeCN (28 mL) and piperidine (28 mL) was added and stirred at room temperature for 30 minutes. After confirming that the starting material disappeared on TLC, the solution was concentrated under reduced pressure and separated by column with ethyl acetate:hexane (3:7) to obtain compound 1-4 (2.2 g) in a white solid state.

[0092] MS: [M+H] + = 381

[0093]

[0094] Step 5) Preparation of 1,2-Dibenzyl 5-(tert-butyl)(1S,3aS,6aR)-tetrahydropyrrolo[3,4-c]pyrrole-1,2,5(1H,3H)-tricarboxylate

[0095] Compound 1-4 (2.2 g) was dissolved in DCM (29 mL), and triethylamine (1.6 mL), Boc2O (2 mL), and DMAP (70 mg) were added and stirred at room temperature. After confirming that the starting material disappeared on TLC, extraction was performed with sat. NH4Cl / DCM. The extracted organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure, and separated by column with ethyl acetate:hexane (1:4) to obtain compound 1-5 (2.6 g) in a white solid state.

[0096] MS: [M+H] + = 481

[0097] 1 H NMR (500 MHz, CDCl3) δ 7.41 - 7.19 (m, 10H), 5.33 - 4.98 (m, 4H), 4.29 (dd,J= 47.1, 3.4 Hz, 1H), 3.84 (q,J= 9.0 Hz, 1H), 3.72 - 3.62 (m, 1H), 3.56 - 3.16 (m, 4H), 2.96 - 2.79 (m, 2H), 1.44 (s, 9H).

[0098]

[0099] Step 6) Preparation of rac-1,2-dibenzyl 5-(tert-butyl)(1R,3aS,6aR)-1-((E)-but-2-en-1-yl)tetrahydropyrrolo[3,4-c]pyrrole-1,2,5(1H,3H)-tricarboxylate

[0100] Compound 1-5 (2.6 g) was dissolved in tetrahydrofuran (55 mL), and the temperature was lowered to 0°C. Then, KHMDS (1 M in THF; 13.7 mL) was slowly added dropwise. Under the same temperature conditions, crotyl bromide (1.4 mL) was added, and the temperature was slowly increased to room temperature and stirred for 30 minutes. After confirming that the starting material disappeared on TLC, sat. NH4Cl was added at 0°C and extracted with ethyl acetate. The extracted organic layer was dried over anhydrous MgSO4, concentrated under reduced pressure, and separated by column with ethyl acetate:hexane (1:4) to obtain compound 1-6 (2.4 g) in a white solid state.

[0101] MS: [M+H] + = 535

[0102] 1H NMR (500 MHz, CDCl3) δ 7.44 - 7.19 (m, 10H), 5.70 - 5.37 (m, 1H), 5.19 - 4.78 (m, 3H), 3.79 - 3.65 (m, 1H), 3.61 - 3.33 (m, 3H), 3.24 (q,J= 12.3 Hz, 1H), 2.99 - 2.78 (m, 2H), 1.61 (d,J= 20.9 Hz, 4H), 1.43 (s, 9H).

[0103]

[0104] Step 7) Preparation of rac-1,2-dibenzyl 5-(tert-butyl)(1R,3aS,6aR)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)tetrahydropyrrolo[3,4-c]pyrrole-1,2,5(1H,3H)-tricarboxylate

[0105] Compound 1-6 (720 mg), [Ir(cod)Cl]2 (181 mg), and dppe (214 mg) were dissolved in DCM (6.8 mL), and pinacolborane (0.49 mL) was slowly added dropwise under nitrogen at room temperature. After stirring overnight and confirming that the starting material had disappeared on TLC, the solution was concentrated under reduced pressure and separated by column with ethyl acetate:hexane (1:4) to obtain compound 1-7 (396 mg) as a white solid.

[0106] MS: [M+H] + = 663

[0107]

[0108] Step 8) Preparation of rac-(3aS,6aR)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid

[0109] 6N-HCl aqueous solution (2 mL) was added to compounds 1-7 (100 mg), and the reaction was completed by refluxing for 3 hours. After cooling to room temperature, the solution was washed twice with MC, and the aqueous solution was concentrated and dried under reduced pressure to obtain compound 1 (35 mg).

[0110] MS: [M+H] += 257

[0111] 1 H NMR (500 MHz, MeOD) δ 3.7 (m, 3H), 3.5-3.4 (m, 2H), 3.3 (m, 2H), 3.2 (m, 1H), 2.1 (m, 1H), 2.0 (m, 1H), 1.4 (m, 3H), 1.3 (m, 1H), 0.8 (m, 2H)

[0112]

[0113] Example 2: Preparation of rac-(3aR,6aR)-1-(4-voronobutyl)-5-methyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0114]

[0115]

[0116] Step 1) Preparation of rac-dibenzyl (3aS,6aR)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)hexahydropyrrolo[3,4-c]pyrrole-1,2(1H)-dicarboxylic acid hydrochloride

[0117]

[0118] Compound 1-7 (11 mg) was dissolved in 4N HCl (in 1,4-dioxane; 0.2 mL) and heated to 50°C and stirred for 30 minutes. After confirming that the starting material disappeared on MS, compound 2-1 (10 mg), a yellow oil, was obtained by concentrating under reduced pressure.

[0119] MS: [M+H] + = 563

[0120]

[0121] Step 2) Preparation of rac-(3aR,6aR)-1-(4-voronobutyl)-5-methyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0122]

[0123] Compound 2-1 (10 mg) was dissolved in tetrahydrofuran (0.2 mL), then formaldehyde (2.1 μL) and sodium triacetoxyborohydride (5.4 mg) were added, and the reaction was completed by stirring at room temperature for 3 hours. After concentrating the reaction mixture, it was separated by column with ethyl acetate (100%), and Compound 2 (2.2 mg) was obtained in the same manner as in Step 8 of Example 1.

[0124] MS: [M+H] + = 271

[0125] 1 H NMR (500 MHz, DO) δ 3.89 (ddd,J= 69.6, 12.1, 8.2 Hz, 1H), 3.70 - 3.50 (m, 2H), 3.47 - 3.33 (m, 2H), 3.30 - 3.13 (m, 1H), 3.02 (m,J= 12.4, 8.1 Hz, 1H), 2.96 - 2.80 (m, 4H), 1.98 (ddd,J= 15.8, 12.1, 5.0 Hz, 1H), 1.82 (ddd,J= 14.9, 11.9, 4.1 Hz, 1H), 1.40 - 1.24 (m, 3H), 1.17 (m,J= 12.7, 7.2 Hz, 1H), 0.70 (dt,J= 7.6, 4.5 Hz, 2H).

[0126]

[0127] Example 3: Preparation of rac-(3aR,6aR)-1-(4-voronobutyl)-5-ethyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0128]

[0129] Compound 2-1 (50 mg) was dissolved in acetonitrile (0.5 mL), potassium carbonate (3 equivalents) and ethyl iodide (3 equivalents) were added, and the reaction was completed by stirring at 50°C for 5 hours. Water and ethyl acetate were added to the reaction mixture to separate the organic layer. After concentration, the mixture was separated by column separation with methanol:dichloromethane (1:9), and Compound 3 (9.4 mg) was obtained in the same manner as in Step 8 of Example 1.

[0130] MS: [M+H] + = 285

[0131] 1 H NMR (500 MHz, MeOD) δ 4.0 (m, 2H), 3.8 (m, 2H), 3.6 (m, 4H), 2.1-2.0 (m, 2H), 1.6 (m, 1H), 1.5 (m, 3H), 1.4 (m, 5H), 0.8 (m, 2H)

[0132]

[0133] Example 4: Preparation of rac-(3aR,6aR)-1-(4-voronobutyl)-5-propyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0134]

[0135] Compound 2-1 (50 mg) was dissolved in acetonitrile (0.5 mL), potassium carbonate (3 equivalents) and propyl iodide (3 equivalents) were added, and the reaction was completed by stirring at 50°C for 5 hours. Water and ethyl acetate were added to the reaction mixture to separate the organic layer. After concentration, the mixture was separated by column separation with methanol:dichloromethane (1:9), and Compound 4 (12.5 mg) was obtained in the same manner as in Step 8 of Example 1.

[0136] MS: [M+H] + = 299

[0137] 1H NMR (500 MHz, MeOD) δ 4.0 (m, 2H), 3.8 (m, 2H), 3.6 (m, 4H), 2.1-2.0 (m, 1H), 1.6 (m, 1H), 1.5 (m, 3H), 1.4 (m, 5H), 1.0 (m, 3H), 0.8 (m, 2H)

[0138]

[0139] Example 5: Preparation of rac-(3aR,6aR)-1-(4-voronobutyl)-5-isopropyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0140]

[0141] Compound 2-1 (58 mg) was dissolved in acetonitrile (0.5 mL), potassium carbonate (3 equivalents) and isopropyl iodide (3 equivalents) were added, and the reaction was completed by stirring at 50°C for 5 hours. Water and ethyl acetate were added to the reaction mixture to separate the organic layer. After concentration, the mixture was separated by column separation with methanol:dichloromethane (1:9), and Compound 5 (9.4 mg) was obtained in the same manner as in Step 8 of Example 1.

[0142] MS: [M+H] + = 299

[0143] 1 H NMR (500 MHz, D2O) δ 3.87 (m,J= 55.1, 12.1, 8.0 Hz, 2H), 3.70 - 3.25 (m, 4H), 3.20 - 2.86 (m, 3H), 1.99 (ddd,J= 16.1, 11.5, 4.5 Hz, 1H), 1.83 (ddd,J= 14.6, 11.7, 3.9 Hz, 1H), 1.38 - 1.15 (m, 10H), 0.75 - 0.63 (m, 2H).

[0144]

[0145] Example 6: Preparation of rac-(3aR,6aR)-5-(3-aminopropyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid trihydrochloride

[0146]

[0147] Compound 2-1 (84 mg) was dissolved in dimethylformamide (0.3 mL), then tert-butyl (3-bromopropyl)carbamate (40 mg), potassium carbonate (23 mg), and diisopropylethylamine (48.9 μL) were added, and the reaction was completed by stirring overnight at 60°C. Water and ethyl acetate were added to the reaction mixture to separate the organic layer. After concentration, column separation with methanol:dichloromethane (1:9) was performed, and Compound 6 (33.7 mg) was obtained in the same manner as in Step 8 of Example 1.

[0148] MS: [M+H] + = 314

[0149] 1 H NMR (500 MHz, D2O) δ 4.08 - 3.86 (m, 1H), 3.68 - 3.57 (m, 1H), 3.48 - 3.19 (m, 6H), 2.99 (m,J= 7.8 Hz, 4H), 2.13 - 1.76 (m, 4H), 1.36 - 1.13 (m, 4H), 0.73 - 0.65 (m, 2H).

[0150]

[0151] Example 7: Preparation of rac-(3aR,6aR)-5-(L-alanyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0152]

[0153]

[0154] Step 1) Preparation of rac-dibenzyl (3aS,6aR)-5-((tert-butoxycarbonyl)-L-alanyl)-1-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)hexahydropyrrolo[3,4-c]pyrrole-1,2(1H)-dicarboxylate

[0155]

[0156] Compound 2-1 (50 mg) was dissolved in dimethylformamide (0.5 mL), and HATU (1.2 equivalents) was added and stirred for 10 minutes. In a separate reactor, 2,5-dioxopyrrolidine-1-yl (tert-butoxycarbonyl)-L-alaniate (1.2 equivalents) and diisopropylamine (3 equivalents) were dissolved in dimethylformamide (0.5 mL) and stirred for 10 minutes. The product was added to the reactor in which Compound 2-1 had previously been reacted. The reaction mixture was stirred at 20 to 30°C for 3 to 5 hours to complete the reaction. Water and ethyl acetate were added to the reaction mixture to separate the organic layer. After concentration, Compound 7-1 (27.6 mg) was obtained by column separation with hexane and ethyl acetate.

[0157] MS: [M+H] + = 734

[0158]

[0159] Step 2) Preparation of rac-(4-((3aS,6aR)-1,2-bis((benzyloxy)carbonyl)-5-((tert-butoxycarbonyl)-L-alanyl)octahydropyrrolo[3,4-c]pyrrolo-1-yl)butyl)boronic acid.

[0160]

[0161] Compound 7-1 (27 mg) was dissolved in acetone (0.5 mL), methylboronic acid (10 equivalents) and 1N-NaOH (1 equivalent) were added, and the reaction was completed by stirring at room temperature for 4 hours. The mixture was concentrated and extracted with a saturated NH4Cl solution and ethyl acetate. After concentrating the organic layer, compound 7-2 (10 mg) was obtained by column separation with methylene chloride:methanol (20:1).

[0162] MS: [M+H] + = 652

[0163]

[0164] Step 3) Preparation of rac-(3aR,6aR)-5-(L-alanyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0165]

[0166] Compound 7-2 (10 mg) was dissolved in methanol, 10% Pd / C was added, and the reaction was completed after 1 hour by bubbling with hydrogen gas. The reaction solution was filtered through celite and concentrated. After dissolving in methanol, 4N-HCl (in dioxane; 10 equivalents) was added and the reaction was completed by stirring at 20 to 30°C. After concentrating the reaction solution, compound 7 (5 mg) was obtained by drying under reduced pressure.

[0167] MS: [M+H] + = 328

[0168] 1 H NMR (500 MHz, MeOD) δ 4.2 (m, 2H), 3.9-3.7 (m, 3H), 3.7-3.6 (m, 2H), 3.5 (m, 1H), 3.4-3.3 (m, 2H), 2.2 (m, 1H), 2.0 (m, 1H), 1.5 (m, 6H), 1.4 (m, 1H), 0.8 (m, 2H)

[0169]

[0170] Example 8: Preparation of rac-(3aR,6aR)-5-(L-phenylalanyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0171]

[0172] Compound 8 (12 mg) was obtained in the same manner as in Example 7, except that compound 7-1 (50 mg) and 2,5-dioxopyrrolidine-1-yl (tert-butoxycarbonyl)-L-phenylalanineate (1.2 equivalents) were used.

[0173] MS: [M+H] + = 404

[0174] 1 H NMR (500 MHz, MeOD) δ 7.4-7.3 (m, 5H), 4.4 (m, 1H), 4.0 (m, 0.5H), 3.7 (m, 0.5H), 3.6 (m, 1H), 3.4-3.35 (m, 1H), 3.2 (m, 1H), 3.1 (m, 3H), 2.6 (m, 1H), 2.2 (m, 0.5H), 2.1 (m, 0.5H), 1.9 (m, 0.5H), 1.8 (m, 0.5H), 1.7 (m, 2H), 1.5-1.4 (m, 3H), 1.3 (m, 1H), 0.8 (m, 2H)

[0175]

[0176] Example 9: Preparation of rac-(3aR,6aR)-5-(L-lucyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0177]

[0178] Compound 9 (14 mg) was obtained in the same manner as in Example 7, except that compound 7-1 (48 mg) and 2,5-dioxopyrrolidine-1-yl (tert-butoxycarbonyl)-L-lucinate (39.5 mg) were used.

[0179] MS: [M+H] + = 370

[0180] 1 H NMR (500 MHz, DMSO) δ 4.03 - 3.00 (m, 7H), 2.02 - 1.12 (m, 8H), 0.92 (dt,J= 12.6, 6.2 Hz, 9H), 0.60 (t,J= 7.7 Hz, 2H).

[0181]

[0182] Example 10: Preparation of rac-(3aR,6aR)-5-((2S)-2-amino-3-methylpentanoyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0183]

[0184] Compound 10 (8.4 mg) was obtained in the same manner as in Example 7, except that compound 7-1 (65 mg) and 2,5-dioxopyrrolidine-1-yl (tert-butoxycarbonyl)-L-isolucinate (53.5 mg) were used.

[0185] MS: [M+H] + = 370

[0186] 1 H NMR (500 MHz, DMSO) δ 3.92 - 3.01 (m, 5H), 2.03 - 1.77 (m, 2H), 1.55 - 0.80 (m, 17H), 0.59 (d,J= 7.5 Hz, 2H).

[0187]

[0188] Example 11: Preparation of rac-(3aR,6aR)-1-(4-voronobutyl)-5-glycyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0189]

[0190] Compound 11 (6 mg) was obtained in the same manner as in Example 7, except that compound 7-1 (134 mg) and 2,5-dioxopyrrolidine-1-yl (tert-butoxycarbonyl)glycinate (53.5 mg) were used.

[0191] MS: [M+H] + = 314

[0192] 1 H NMR (500 MHz, D2O) δ 3.86 - 3.74 (m, 2H), 3.72 - 3.40 (m, 5H), 3.35 - 3.17 (m, 2H), 3.06 (dq,J= 31.2, 7.5 Hz, 1H), 2.04 (ddd,J= 15.9, 12.7, 4.5 Hz, 1H), 1.78 (m,J= 15.6, 12.4, 4.2 Hz, 1H), 1.39 - 1.12 (m, 4H), 0.69 (td,J= 7.6, 2.8 Hz, 2H).

[0193]

[0194] Example 12: Preparation of rac-(3aR,6aR)-5-(L-prolyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid dihydrochloride

[0195]

[0196] Compound 12 (5 mg) was obtained in the same manner as in Example 7, except that compound 7-1 (126 mg) and 2,5-dioxopyrrolidine-1-yl (tert-butoxycarbonyl)-L-prolinate (99 mg) were used.

[0197] MS: [M+H] + = 354

[0198] 1H NMR (500 MHz, DO) δ 4.41 (t,J= 9.1 Hz, 1H), 3.80 - 3.41 (m, 5H), 3.38 - 3.16 (m, 4H), 3.03 (dd,J= 31.2, 7.5 Hz, 1H), 2.47 - 2.36 (m, 1H), 2.10 - 1.71 (m, 5H), 1.41 - 1.11 (m, 4H), 0.68 (m,J= 7.7 Hz, 2H).

[0199]

[0200] Example 13: Preparation of rac-(3aR,6aR)-5-(L-lysyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid trihydrochloride

[0201]

[0202] Compound 7-1 (101 mg) and 2,5-dioxopyrrolidine-1-yl N 2 ,N 6 Compound 13 (5 mg) was obtained in the same manner as in Example 7, except that bis(tert-butoxycarbonyl)-L-lysine (113 mg) was used.

[0203] MS: [M+H] + = 385

[0204] 1 H NMR (500 MHz, D2O) δ 4.16 (m, 1H), 3.89 - 3.48 (m, 5H), 3.36 - 3.20 (m, 2H), 3.10 - 2.87 (m, 3H), 2.03 (m,J= 13.7 Hz, 1H), 1.85 - 1.71 (m, 3H), 1.68 - 1.57 (m, 2H), 1.44 - 1.10 (m, 6H), 0.69 (m,J= 8.1 Hz, 2H).

[0205]

[0206] Experimental Example 1: Arginase Inhibitory Activity Test

[0207] An arginase inhibitory activity test was conducted on the compound according to the present invention.

[0208]

[0209] Specifically, all reagents required for evaluation were diluted in reaction buffer (8 mM NAHPO4, 2 mM KH2PO4, pH 7.5, 137 mM NaCl, 2.7 mM KCl, and 0.05% Tween-20). The compounds were dissolved in DMSO and diluted, and then diluted in reaction buffer to the desired concentration. 10 μL of the compound and 10 μL of 30 nM Arginase-1 were mixed in a clear 96-well plate (SPL) and incubated at room temperature for at least 1 hour. Subsequently, 10 μL of 15 mM L-arginine and 3 mM MnCl2 were added to the plate and reacted at room temperature for at least 30 minutes. Finally, 30 μL of a 1:1 mixture of reagent A (10 mM o-phthaldialdehyde, 0.4% polyoxyethylene lauryl ether, and 1.8 M sulfuric acid in DW) and reagent B (1.3 mM primaquine diphosphate, 0.4% polyoxyethylene lauryl ether, and 3.6 M sulfuric acid in DW) was added to complete the enzymatic reaction, and the mixture was incubated at room temperature for at least 1 hour. After incubation, the absorbance of the plate at 450 nM was measured using a Flexstation3 multi-mode microplate reader (Molecular Devices).

[0210]

[0211] The ΔOD for each reading was obtained by subtracting the background control (BC) reading from all readings, and after setting the ΔOD of the enzyme control (EC) to the maximum value, the % relative Arginase-1 activity of each compound (S) was calculated using the following formula, and the IC of the compounds was calculated using PRISM (GraphPad software). 50 The value was determined.

[0212] Relative Arginase-1 activity (%) = (ΔOD of S / ΔOD of EC)*100

[0213]

[0214] The efficacy of the compounds is listed in Table 1 below on a scale from A to C. The efficacy value for A is an IC50 of less than 100 nM. 50 Refers to a compound of the present invention having a value, and a compound with an efficacy value of B has an IC in the range of 100 nM to 1,000 nM. 50 Indicates a value, ICs exceeding 1000 nM 50 The efficacy value of C was assigned to compounds having the value.

[0215] Target IC 50 Example 1 (Compound 1) A Example 2 (Compound 2) A Example 3 (Compound 3) A Example 4 (Compound 4) A Example 5 (Compound 5) A Example 6 (Compound 6) A Example 7 (Compound 7) B Example 8 (Compound 8) B Example 9 (Compound 9) B Example 10 (Compound 10) B Example 11 (Compound 11) B Example 12 (Compound 12) B Example 13 (Compound 13) B

[0216] Experimental Example 2: PBMC Activity Evaluation Test

[0217] Primary Peripheral Blood Mononuclear Cells (PBMC) activity tests were conducted on the compound according to the present invention. After inhibiting T cell differentiation induced by α-CD3 and α-CD28 antibodies via synthetic arginase1, it was confirmed that the compound inhibits arginase1 to induce T cell differentiation and proliferation. To evaluate T cell proliferation, the CellTiter-Glo® Luminescent Cell Viability (Promega, Catalog #G7571) evaluation method was used, which can confirm cell proliferation by quantifying ATP present in the cells.

[0218]

[0219] Specifically, an α-CD3 antibody was coated onto a clear 96-well microplate. 1 x 10⁶ 5 PBMC cells were placed in each well. RPMI media (10% FBS, 1% penicillin / streptomycin) was used as the cell culture medium. α-CD28 antibody and synthetic arginase 1 were added to each well. The compounds prepared in the example were treated at various concentrations (2, 1, 0.5, 0.25, 0.125, 0.0625, 0.03125 μM). After 72 hours of incubation, 50 μL of cells from each well were transferred to a white 96-well microplate, and 50 μL of CellTiter-Glo® solution was mixed in. After reacting at room temperature in the dark for 10 minutes, luminescene was measured. The measurement results for each compound were calculated using Excel, and EC 50 The value was calculated using GraphPad Prism software.

[0220]

[0221] The efficacy of the compounds is listed in Table 2 below on a scale from A to C. The efficacy value for A is an EC of less than 100 nM.50 Refers to the compound of the present invention having a value, and the compound with an efficacy value of B has an EC in the range of 100 nM to 1,000 nM. 50 Represents a value, EC exceeding 1000 nM 50 The efficacy value of C was assigned to compounds having the value.

[0222] Target EC 50 Example 1 (Compound 1) A Example 3 (Compound 3) A Example 5 (Compound 5) B Example 6 (Compound 6) B

Claims

1. A compound represented by the following chemical formula 1, or a pharmaceutically acceptable salt thereof: [Chemical Formula 1] In the above chemical formula 1, R is R1, -(CO)-CH(NH2)-R2, or -(CO)-R3, and R1 is hydrogen, C 1-4 Alkyl, C 1-4 haloalkyl, amino, or -(C 1-4 It is an alkylene)amino, and R2 is hydrogen, C 1-4 Alkyl, -(C 1-4 Alkylene)amino, phenyl, or benzyl, and R3 is a 5-membered or 6-membered heterocycloalkyl containing at least one of N, O, and S.

2. In Paragraph 1, R1 is hydrogen, methyl, ethyl, propyl, isopropyl, trifluoromethyl, 2,2,2-trifluoroethyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, or 4-aminobutyl, A compound, or a pharmaceutically acceptable salt thereof.

3. In Paragraph 1, R2 is hydrogen, methyl, ethyl, propyl, isopropyl, 1-butyl, 2-butyl, isobutyl, tertbutyl, aminomethyl, 2-aminoethyl, 3-aminopropyl, 4-aminobutyl, phenyl, or benzyl, A compound, or a pharmaceutically acceptable salt thereof.

4. In Paragraph 1, R3 is pyrrolidinyl, A compound, or a pharmaceutically acceptable salt thereof.

5. In Paragraph 1, The compound represented by the above chemical formula 1 is, 1) 1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 2) 1-(4-voronobutyl)-5-methyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 3) 1-(4-voronobutyl)-5-ethyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 4) 1-(4-voronobutyl)-5-propyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 5) 1-(4-voronobutyl)-5-isopropyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 6) 5-(3-aminopropyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 7) 5-(L-alanyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 8) 5-(L-phenylalanyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 9) 5-(L-lucyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 10) 5-((2S)-2-amino-3-methylpentanoyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 11) 1-(4-voronobutyl)-5-glycyloctahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 12) 5-(L-prolyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, 13) 5-(L-lysyl)-1-(4-voronobutyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid, and 14) 1-(4-voronobutyl)-5-(2,2,2-trifluoroethyl)octahydropyrrolo[3,4-c]pyrrole-1-carboxylic acid One of which is selected from the group consisting of A compound, or a pharmaceutically acceptable salt thereof.

6. A pharmaceutical composition for the prevention or treatment of cancer or tumor, comprising a compound of any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof.

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