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 containing the compound or pharmaceutically acceptable salt thereof can be effectively 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 according to the present invention 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 or 3-aminopropyl.

[0021]

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

[0023]

[0024] Preferably, R3 is pyrrolidinyl.

[0025]

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

[0027] 1) 7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid,

[0028] 2) 7-(4-voronobutyl)-2-glycyl-2,6-diazaspiro[3,4]octane-7-carboxylic acid,

[0029] 3) 2-(L-alanyl)-7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid, and

[0030] 4) 2-(3-aminopropyl)-7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid.

[0031]

[0032] 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.

[0033]

[0034] Other pharmaceutically unacceptable salts or solvates of compounds 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.

[0035]

[0036] 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.

[0037]

[0038] 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.

[0039]

[0040] 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:

[0041] [Reaction Equation 1]

[0042]

[0043] 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.

[0044]

[0045] 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.

[0046]

[0047] 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.

[0048]

[0049] 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.

[0050]

[0051] 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.

[0052]

[0053] 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.

[0054]

[0055] 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.

[0056]

[0057] 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.

[0058]

[0059] 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.

[0060]

[0061] 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.

[0062]

[0063] [Example]

[0064] Example 1: Preparation of 7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid dihydrochloride

[0065]

[0066] Step 1) Preparation of 6-benzyl 2-(tert-butyl) 7-ethyl 2,6-diazaspiro[3,4]octane-2,6,7-tricarboxylate

[0067]

[0068] 2-(tert-butyl) 7-ethyl 2,6-diazaspiro[3,4]octane-2,7-dicarboxylate (100 mg) was dissolved in dichloromethane (1.5 mL), then triethylamine (98.1 μL) and benzyl chloroformate (60.2 μL) were slowly added dropwise, and the reaction was completed by stirring at room temperature for 30 minutes. The mixture was separated into layers using ethyl acetate and a saturated aqueous solution of ammonium chloride, and the organic layer was separated. The aqueous layer was extracted twice using ethyl acetate, and the organic layer was separated. The organic layer was dried with MgSO4 and then concentrated. Compound 1-1 (67 mg, yield 45%) was obtained by column separation using ethyl acetate (in 30% Hexane).

[0069] MS: [M+H] + = 419

[0070] 1H NMR (500 MHz, CDCl3) δ 7.39 - 7.27 (m, 5H), 5.20 - 5.01 (m, 2H), 4.38 (ddd,J= 27.3, 8.7, 3.9 Hz, 1H), 4.20 (q,J= 7.2 Hz, 1H), 4.09 - 3.97 (m, 1H), 3.94 - 3.78 (m, 4H), 3.74 (dd,J= 9.0, 2.4 Hz, 1H), 3.64 (dd,J= 19.4, 11.0 Hz, 1H), 2.40 (ddd,J= 17.2, 13.2, 8.8 Hz, 1H), 2.25 (ddd,J= 12.4, 6.6, 3.9 Hz, 1H), 1.43 (d,J= 3.3 Hz, 9H), 1.20 (dt,J= 68.5, 7.1 Hz, 3H)

[0071]

[0072] Step 2) Preparation of 6-((benzyloxy)carbonyl)-2-(tert-butoxycarbonyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid

[0073]

[0074] Compound 1-1 (67 mg) was dissolved in an ethanol / water mixed solution (5:3, 0.2 mL), lithium hydroxide (13.4 mg) was added, and the reaction was completed by stirring at room temperature for 1 hour. The layers were separated using ethyl acetate and a saturated aqueous solution of ammonium chloride, and the organic layer was separated. The aqueous layer was extracted twice using ethyl acetate, and the organic layer was separated. The organic layer was dried and concentrated with MgSO4, and the product was used directly in Step 3 below.

[0075] MS: [M+H] + = 257

[0076]

[0077] Step 3) Preparation of 6,7-Dibenzyl 2-(tert-butyl) 2,6-diazaspiro[3,4]octane-2,6,7-tricarboxylate

[0078]

[0079] Compound 1-2 (30 mg), sodium iodide (2.3 mg), and potassium carbonate (12.7 mg) were dissolved in N,N-dimethylformamide (0.4 mL), then benzyl bromide (11 μL) was added and the reaction was completed by stirring at room temperature for 3 hours. The mixture was filtered with ethyl acetate using Celite and then concentrated. Compound 1-3 (28 mg, yield 76%) was obtained by column separation using ethyl acetate (in hexane 20%).

[0080] MS: [M+H] + = 347

[0081] 1 H NMR (500 MHz, CDCl3) δ 7.61 - 7.09 (m, 10H), 5.27 - 5.10 (m, 2H), 5.07 - 4.98 (m, 2H), 4.44 (ddd,J= 37.1, 8.8, 3.9 Hz, 1H), 3.94 - 3.76 (m, 4H), 3.69 - 3.56 (m, 2H), 2.39 (ddd,J= 19.6, 13.2, 8.8 Hz, 1H), 2.22 (td,J= 13.4, 4.0 Hz, 1H), 1.42 (d,J= 3.9 Hz, 9H).

[0082]

[0083] Step 4) Preparation of 6,7-Dibenzyl 2-(tert-butyl)(E)-7-(butene-2-1-yl)-2,6-diazaspiro[3,4]octane-2,6,7-tricarboxylate

[0084]

[0085] Compound 1-3 (73 mg) was dissolved in tetrahydrofuran (1.5 mL), the temperature was lowered to 0 °C, and 1 N lithium bis(trimethylsilyl)amide (0.29 mL) was slowly added dropwise under nitrogen conditions. After stirring for 20 minutes at the same temperature, crotyl bromide (51.3 μL) was added, and the reaction was completed by stirring for 1 hour at the same temperature. The mixture was separated into layers using ethyl acetate and a saturated aqueous solution of ammonium chloride, and the organic layer was separated. The aqueous layer was extracted twice using ethyl acetate, and the organic layer was separated. The organic layer was dried with MgSO4 and then concentrated. Compound 1-4 (65 mg, yield 80%) was obtained by column separation using ethyl acetate (in hexane 20%).

[0086] MS: [M+H] + = 491

[0087] 1 H NMR (500 MHz, CDCl3) δ 7.40 - 7.17 (m, 10H), 5.35 - 4.99 (m, 3H), 3.96 - 3.43 (m, 6H), 3.09 (dd,J= 14.8, 6.3 Hz, 1H), 2.57 (dd,J= 14.2, 8.5 Hz, 1H), 2.34 - 2.15 (m, 4H), 1.64 (t,J= 6.5 Hz, 3H), 1.41 (s, 9H).

[0088]

[0089] Step 5) Preparation of 6,7-Dibenzyl 2-(tert-butyl) 7-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)-2,6-diazaspiro[3,4]octane-2,6,7-tricarboxylate

[0090]

[0091] Compound 1-4 (49 mg), chloro(1,5)-cyclooctadiene)iridium(I) dimer (12.3 mg), and 1,2-bis(diphenylphosphino)ethane (14.6 mg) were dissolved in dichloromethane (1 mL), and then 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (33.2 μL) was added dropwise under nitrogen conditions and stirred overnight at room temperature. After concentrating the reaction mixture, compound 1-5 (45 mg, yield 74%) was obtained by column separation using ethyl acetate (in hexane 30%).

[0092] MS: [M+H] + = 619

[0093] 1 H NMR (500 MHz, CDCl3) δ 7.41 - 7.17 (m, 10H), 5.19 - 4.85 (m, 2H), 4.10 - 3.95 (m, 1H), 3.85 - 3.66 (m, 3H), 3.50 (dd,J= 20.3, 10.5 Hz, 2H), 2.40 - 2.07 (m, 3H), 1.87 (dt,J= 57.8, 13.5 Hz, 1H), 1.59 (s, 2H), 1.41 (s, 9H), 1.24 (m,J= 16.0 Hz, 16H), 0.73 (dt,J= 33.5, 7.8 Hz, 2H).

[0094]

[0095] Step 6) Preparation of 2-(tert-butoxycarbonyl)-7-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid

[0096]

[0097] Compound 1-5 (45 mg) and 10% Pd / C were dissolved in methanol (0.5 mL), and the reaction was completed by stirring under hydrogen conditions for 3 hours. The reaction mixture was filtered with methanol using Celite to remove Pd / C and concentrated, and the product was used directly in step 7 below.

[0098] MS: [M+H] + = 439

[0099]

[0100] Step 7) Preparation of 7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid dihydrochloride

[0101]

[0102] The product 1-6 (28 mg) from step 6 above was dissolved in a 1 N aqueous hydrochloric acid solution (1 mL), and then hexane (1 mL) and phenylboronic acid (8 mg) were added in sequence and stirred vigorously for 30 minutes. After removing the hexane layer, the aqueous solution was washed three times with diethyl ether and the aqueous layer was concentrated. The product was dissolved again in a 1 N aqueous hydrochloric acid solution (1 mL), and the reaction was completed by stirring at 40 °C for 30 minutes. The aqueous solution was washed three times again with diethyl ether and the aqueous layer was concentrated, and then dried under reduced pressure to obtain compound 1 (17 mg, yield 81%).

[0103] MS: [M+H] + = 257

[0104] 1 H NMR (500 MHz, DO) δ 4.22 (d,J= 11.6 Hz, 1H), 4.11 (dd,J= 11.4, 8.8 Hz, 2H), 3.96 (d,J= 11.4 Hz, 1H), 3.78 (d,J= 12.9 Hz, 1H), 3.58 (d,J= 12.9 Hz, 1H), 2.87 (d,J= 14.1 Hz, 1H), 2.28 (d,J= 14.1 Hz, 1H), 2.05 (ddd,J= 14.5, 12.1, 4.7 Hz, 1H), 1.70 (ddd,J= 14.5, 12.0, 4.4 Hz, 1H), 1.38 - 1.29 (m, 2H), 1.29 - 1.09 (m, 2H), 0.76 - 0.63 (m, 2H).

[0105]

[0106] Example 2: Preparation of 7-(4-voronobutyl)-2-glycyl-2,6-diazaspiro[3,4]octane-7-carboxylic acid dihydrochloride

[0107]

[0108] Step 1) Preparation of dibenzyl 7-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)-2,6-diazaspiro[3,4]octane-6,7-dicarboxylate hydrochloride

[0109]

[0110] Compound 1-5 (362 mg) was dissolved in 4 N hydrochloric acid (2 mL), and the reaction was completed by stirring at 50 °C for 30 minutes. The concentrated product was used directly in Step 2 below.

[0111] MS: [M+H] + = 519

[0112]

[0113] Step 2) Preparation of dibenzyl 2-((tert-butoxycarbonyl)glycyl)-7-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)-2,6-diazaspiro[3,4]octane-6,7-dicarboxylate

[0114]

[0115] Compound 2-1 (107 mg) was dissolved in dichloromethane (0.9 mL), then triethylamine (75 μL) and 2,5-dioxopyrrolidine-1-yl (tert-butoxycarbonyl)glycinate (73 mg) were added, and the reaction was completed by stirring overnight. After concentrating the reaction mixture, compound 2-2 (71 mg, yield 55%) was obtained by column separation using ethyl acetate (100%).

[0116] MS: [M+H] + = 676

[0117] 1H NMR (500 MHz, CDCl3) δ 7.40 - 7.20 (m, 10H), 5.27 - 4.76 (m, 2H), 4.06 - 3.82 (m, 3H), 3.78 - 3.65 (m, 2H), 3.48 (d,J= 12.4 Hz, 3H), 2.40 - 2.08 (m, 3H), 1.87 (d,J= 55.7 Hz, 1H), 1.60 (s, 2H), 1.44 (d,J= 8.7 Hz, 9H), 1.37 - 1.05 (m, 16H), 0.74 (dd,J= 24.1, 16.1 Hz, 2H).

[0118]

[0119] Step 3) Preparation of 7-(4-voronobutyl)-2-glycyl-2,6-diazaspiro[3,4]octane-7-carboxylic acid dihydrochloride

[0120]

[0121] Compound 2 was obtained in the same manner as steps 6 and 7 of Example 1, except that compound 2-2 (71 mg) was used instead of compound 1-5 in step 6 of Example 1.

[0122] MS: [M+H] + = 314

[0123] 1H NMR (500 MHz, D2O) δ 4.24 (m,J= 9.7 Hz, 1H), 4.15 - 4.03 (m, 1H), 3.99 - 3.89 (m, 1H), 3.79 - 3.64 (m, 3H), 3.58 - 3.50 (m, 1H), 3.48 - 3.34 (m, 1H), 2.76 (dd,J= 13.9, 2.5 Hz, 1H), 2.23 (dd,J= 14.0, 2.3 Hz, 1H), 2.09 - 1.98 (m, 1H), 1.70 (ddd,J= 14.4, 12.1, 4.5 Hz, 1H), 1.33 (dq,J= 14.7, 6.5 Hz, 2H), 1.19 (dq,J= 50.9, 5.8 Hz, 2H), 0.76 - 0.63 (m, 2H).

[0124]

[0125] Example 3: Preparation of 2-(L-alanyl)-7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid dihydrochloride

[0126]

[0127] Compound 3 was obtained in the same manner as steps 2 and 3 of Example 2, except that in step 2 of Example 2, 2,5-dioxopyrrolidine-1-yl((benzyloxy)carbonyl)-L-alaninate was used instead of 2,5-dioxopyrrolidine-1-yl(tert-butoxycarbonyl)glycinate.

[0128] MS: [M+H] + = 328

[0129] 1H NMR (500 MHz, D2O) δ 4.38 - 3.89 (m, 3H), 3.81 - 3.70 (m, 1H), 3.64 - 3.48 (m, 1H), 2.82 - 2.70 (m, 1H), 2.25 (ddd,J= 14.0, 6.7, 4.0 Hz, 1H), 2.10 - 1.92 (m, 1H), 1.71 (ddd,J= 14.9, 12.1, 4.4 Hz, 1H), 1.50 - 1.41 (m, 1H), 1.36 (dp,J= 8.6, 5.0 Hz, 5H), 1.20 (ddt,J= 48.5, 11.7, 5.9 Hz, 2H), 0.76 - 0.63 (m, 2H).

[0130]

[0131] Example 4: Preparation of 2-(3-aminopropyl)-7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid trihydrochloride

[0132]

[0133] Step 1) Preparation of dibenzyl 2-(3-((tert-butoxycarbonyl)amino)propyl)-7-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)butyl)-2,6-diazaspiro[3,4]octane-6,7-dicarboxylate

[0134]

[0135] Compound 2-1 (382 mg) was dissolved in N,N-dimethylformamide (1.3 mL), and then N,N-diisopropylethylamine (0.22 mL), tert-butyl(2-bromoethyl)carbamate (182 mg), and potassium carbonate (106 mg) were added in sequence and stirred at 60 °C for 2 hours. The reaction mixture was filtered and concentrated using Celite, and then column-separated with methanol (in CH2Cl2 10%) solvent to obtain Compound 4-1 (244 mg, yield 53%).

[0136] MS: [M+H] + = 720

[0137]

[0138] Step 2) Preparation of 2-(3-aminopropyl)-7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid trihydrochloride

[0139]

[0140] Compound 4 was obtained in the same manner as steps 6 and 7 of Example 1, except that compound 4-1 (244 mg) was used instead of compound 1-5 in step 6 of Example 1.

[0141] MS: [M+H] + = 314

[0142] 1 H NMR (500 MHz, DO) δ 4.44 - 4.20 (m, 2H), 4.15 - 4.02 (m, 2H), 3.80 (dd,J= 50.6, 13.0 Hz, 1H), 3.69 - 3.54 (m, 1H), 3.28 (dt,J= 16.7, 8.0 Hz, 2H), 2.98 (t,J= 7.8 Hz, 2H), 2.86 (dd,J= 45.0, 14.2 Hz, 1H), 2.29 (dd,J= 14.1, 3.6 Hz, 1H), 2.12 - 1.98 (m, 1H), 1.87 (m,J= 12.5, 9.2, 5.2 Hz, 2H), 1.70 (m,J= 15.6, 11.8, 4.3 Hz, 1H), 1.33 (m,J= 7.5 Hz, 2H), 1.20 (m,J= 46.1, 13.8, 7.7 Hz, 2H), 0.69 (td,J= 7.7, 2.2 Hz, 2H).

[0143]

[0144] Experimental Example 1: Arginase Inhibitory Activity Test

[0145] An arginase inhibitory activity test was conducted on the compound according to the present invention.

[0146]

[0147] 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).

[0148]

[0149] 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.

[0150] Relative Arginase-1 activity (%) = (ΔOD of S / ΔOD of EC)*100

[0151]

[0152] 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.

[0153]

[0154] Target IC 50 1B2C3C4B

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) 7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid, 2) 7-(4-voronobutyl)-2-glycyl-2,6-diazaspiro[3,4]octane-7-carboxylic acid, 3) 2-(L-alanyl)-7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-carboxylic acid, and 4) 2-(3-aminopropyl)-7-(4-voronobutyl)-2,6-diazaspiro[3,4]octane-7-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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