Compounds for use in the treatment of gastric cancer

Abstract

Formula I: Methods are provided for treating gastric cancer in a subject using compounds and compositions comprising a compound having the formula: JPEG2024543111000033.jpg3062 and pharma- ceutically acceptable salts thereof.

Description

[Technical field]

[0001] Related Applications This application claims the benefit of priority to U.S. Provisional Application No. 63 / 280,540, filed November 17, 2021, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] The American Cancer Society estimates that there will be approximately 26,560 new cases of gastric cancer (also known as stomach cancer) this year, with 11,180 deaths from this type of cancer. Approximately 6 of every 10 people diagnosed with gastric cancer are over the age of 65. Gastric cancer is difficult to treat, and with current therapies, survival rates are low (5-year survival rate is 32% for all stages and 5% for metastatic gastric cancer), representing a clear unmet clinical need. Summary of the Invention

[0003] It has now been found that 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide significantly reduces tumor growth in human xenograft models of gastric cancer. See, for example, Figure 1, where 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide showed more than 70% HS764T tumor growth inhibition (TGI) at 150 mg / kg in nude mice.

[0004] Thus, provided herein is a method of treating gastric cancer using 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide or a pharma- ceutically acceptable salt thereof.

[0005] [ka] Also provided herein is a method for treating gastric cancer using a compound of the formula: 1 , R 2 , R 3 , Z 1 , Z 2 , and p are as defined herein. [Brief description of the drawings]

[0006] [Figure 1] FIG. 1 shows the tumor growth inhibitory effect of 15 days of treatment with 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide on HS746T tumors in nude mice. [Diagram 2] FIG. 1 shows levels of cyclin B1 (CCNB1) and phospho-histone H3 (pHH3) from HS746T xenograft tumors in a xenograft model treated with 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide. [Diagram 3] FIG. 1 shows the tumor growth inhibitory effect of 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide on SNU5 tumors in nude mice for 21 days. [Figure 4] FIG. 1 shows dose-dependent increases in cyclin B1 and pHisH3 in tumors treated with 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] Methods are provided for treating gastric cancer in a subject, comprising administering to the subject a therapeutically effective amount of 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide or a pharma- ceutically acceptable salt thereof.

[0008] 2-(Difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide has the chemical structure shown below and can be synthesized according to the procedure described for compound 126 in U.S. Pat. No. 11,091,447, the entire contents of which are incorporated herein by reference. [ka] 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide has previously been shown to stabilize monoubiquitinated UBE2K in polyubiquitination assays, and is therefore considered, in one aspect, to be an effector of UBE2K. See, for example, FIG. 2 of U.S. Pat. No. 11,091,447. However, recent evidence suggests that 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide may also act as a regulator (e.g., inhibitor) of microtubule formation.

[0009] 2-(Difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide can exist in various tautomeric forms, each of which is expressly included as part of the present invention.

[0010] Also provided is a method of treating gastric cancer in a subject, comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

[0011] There is also provided the use of 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide or a pharma- ceutically acceptable salt thereof for the manufacture of a medicament for treating gastric cancer (e.g., in a subject).

[0012] There is also provided the use of 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide or a pharma- ceutically acceptable salt thereof for treating gastric cancer (e.g., in a subject).

[0013] Also provided is a pharma- ceutically acceptable composition comprising 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide or a pharma- ceutically acceptable salt thereof for treating gastric cancer (e.g., in a subject).

[0014] A therapeutically effective amount of a compound of Formula I: [ka] Also provided is a method of treating gastric cancer in a subject, comprising administering to the subject a compound of the formula: Z 1 and Z 2 are each independently N or CH; X is N or CH; Ring A is phenyl or 5- to 9-membered heteroaryl, each of which is R 5 is optionally substituted with 1 to 3 groups selected from Y is CH2, -CHR a , -CR a R b , or SO, R a and R b are each independently halo, (C1-C6)alkyl, or halo(C1-C6)alkyl, or R a and R b is Ra and R b together with the carbon atom to which it is attached form a 3- to 6-membered cycloalkyl or a 3- to 6-membered heterocyclyl, each of which is optionally substituted with 1 to 3 groups selected from halo, (C1-C6)alkyl, halo(C1-C6)alkyl, (C1-C6)alkoxy, halo(C1-C6)alkoxy, (C1-C6)alkylOH, (C1-C6)alkylO(C1-C6)alkyl, and OH; R 1 is halo(C1-C6)alkyl, halo(C1-C6)alkoxy, or -NR c R d wherein two available hydrogen atoms on the halo(C1-C6)alkyl and halo(C1-C6)alkoxy may, together with the carbon atom to which the halo(C1-C6)alkyl and halo(C1-C6)alkoxy are attached, form a 3- to 6-membered cycloalkyl optionally substituted with one to three groups selected from halo, (C1-C6)alkyl, halo(C1-C6)alkyl, (C1-C6)alkoxy, and halo(C1-C6)alkoxy; R c and R d are each independently hydrogen, (C1-C6)alkyl, halo, (C1-C6)alkyl, (C1-C6)alkylO(C1-C6)alkyl, halo(C1-C6)alkylO(C1-C6)alkyl, (C1-C6)alkyl-O-halo(C1-C6)alkyl, halo(C1-C6)alkyl-O-halo(C1-C6)alkyl, or (C1-C6)alkylOH; or R c and R d is R c and R d together with the nitrogen atom to which it is attached form a 4- to 7-membered heterocyclyl optionally substituted with one to three groups selected from halo, (C1-C6)alkyl, halo(C1-C6)alkyl, (C1-C6)alkoxy, halo(C1-C6)alkoxy, and oxo; R 2 is CN, halo, OH, (C1-C6)alkyl, halo(C1-C6)alkyl, (C1-C6)alkoxy, or halo(C1-C6)alkoxy, or R 1 and R2 is on an adjacent carbon atom, R 1 and R 2 together with the carbon atom to which is attached may form a 5- or 6-membered oxygen-containing heterocyclyl optionally substituted with 1 to 3 groups selected from halo, (C1-C6)alkyl, and halo(C1-C6)alkyl; R 3 is hydrogen, (C1-C6)alkyl, or halo(C1-C6)alkyl; R 4 is CN, halo, OH, (C1-C6)alkyl, halo(C1-C6)alkyl, (C1-C6)alkoxy, halo(C1-C6)alkoxy, -NH(C1-C6)alkyl, -N[(C1-C6)alkyl]2, or 5- or 6-membered heterocyclyl; p is 0 or 1.

[0015] Also provided is a method of treating gastric cancer in a subject, comprising administering to the subject a pharmaceutical composition comprising a therapeutically effective amount of a compound of Formula I, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

[0016] There is also provided the use of a compound of Formula I, or a pharma- ceutically acceptable salt thereof, for the manufacture of a medicament for treating gastric cancer (eg, in a subject).

[0017] There is also provided the use of a compound of Formula I, or a pharma- ceutically acceptable salt thereof, for treating gastric cancer (eg, in a subject).

[0018] Also provided is a pharma- ceutically acceptable composition comprising a compound of Formula I, or a pharma- ceutically acceptable salt thereof, for treating gastric cancer (eg, in a subject).

[0019] In one embodiment, the compound of formula I has formula II or III: [ka] or a pharma- ceutically acceptable salt thereof, where the remaining variables are as described above for formula I.

[0020] In one embodiment, the compound of formula I has formula IV: [ka] or a pharma- ceutically acceptable salt thereof, where the remaining variables are as described above for formula I.

[0021] In one embodiment, R in a compound of formula I, II, III, or IV or a pharma- ceutically acceptable salt thereof 3 is hydrogen, where the remaining variables are as described above for formula I.

[0022] In one embodiment, Y in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, is CH2, SO2, or cyclopropyl, where the remaining variables are as described above for Formula I or any of the preceding embodiments. In the alternative, Y in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, is CH2, where the remaining variables are as described above for Formula I or any of the preceding embodiments.

[0023] In one embodiment, in a compound of formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, Z 1 is N and Z 2 is CH or Z 1 is CH and Z 2 is N or Z 1 and Z 2 Each is CH, where the remaining variables are as described above for Formula I or any of the preceding embodiments. In an alternative embodiment, Z in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, 1 and Z 2 Each is CH, where the remaining variables are as described above for Formula I or any of the preceding embodiments.

[0024] In one embodiment, ring A in a compound of formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, is each R 5wherein the remaining variables are as described above for Formula I or any of the preceding embodiments. In an alternative embodiment, ring A in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, is each selected from R 5 In another alternative, ring A in a compound of formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, is each independently selected from R 5 wherein the remaining variables are as described above for Formula I or any of the preceding embodiments. In another alternative, Ring A in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, is selected from R 5 wherein the remaining variables are as described above for Formula I or any of the preceding embodiments.

[0025] In one embodiment, R in a compound of formula I, II, III, or IV or a pharma- ceutically acceptable salt thereof 1 and R 2 are on adjacent carbon atoms, and R 1 and R 2 together with the carbon atom to which it is attached form a 5-membered oxygen-containing heterocyclyl optionally substituted with 1 or 2 halo, where the remaining variables are as described above for Formula I or any of the preceding embodiments. In an alternative embodiment, R in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, 1 and R 2 are on adjacent carbon atoms, and R 1 and R 2 together with the carbon atom to which it is attached form a dioxolanyl optionally substituted with 1 or 2 halo, where the remainder of the variables are as described above for Formula I or any of the preceding embodiments.

[0026] In one embodiment, R in a compound of formula I, II, III, or IV or a pharma- ceutically acceptable salt thereof 1 is halo(C1-C4)alkyl, halo(C1-C4)alkoxy, or -NR c R d and R c is hydrogen, and R d is halo(C1-C4)alkyl, or R c and R d together form a 4- to 7-membered heterocyclyl optionally substituted with 1 to 3 groups selected from halo, (C1-C4)alkyl, and oxo, where the remaining variables are as described above for Formula I or any of the preceding embodiments. In an alternative embodiment, R in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, 1 is -OCF3, -OCHF2, -OCH2CF3, -CF3, -CH2CF3, -CHF2, piperidinyl, pyrrolidinyl, azapanyl, morpholinyl, thiomorpholinyl, piperazinyl, or azetidinyl, where each of the heterocycles is optionally substituted with 1 to 3 groups selected from halo, (C1-C4)alkyl, and oxo, and the remaining variables are as described above for Formula I or any of the embodiments above.

[0027] In one embodiment, R in a compound of formula I, II, III, or IV or a pharma- ceutically acceptable salt thereof 2 is CN, halo, (C1-C4)alkyl, halo(C1-C4)alkyl, or (C1-C4)alkoxy, where the remaining variables are as described above for Formula I or any of the preceding embodiments. In an alternative embodiment, R in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, 2 is CN or halo, where the remaining variables are as described above for Formula I or any of the preceding embodiments. In another alternative, R in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, 2 is fluoro, where the remaining variables are as described above for Formula I or any of the preceding embodiments.

[0028] In one embodiment, p in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, is 0, where the remaining variables are as described above for Formula I or any of the preceding embodiments.

[0029] In one embodiment, R in a compound of formula I, II, III, or IV or a pharma- ceutically acceptable salt thereof 5 is halo, (C1-C4)alkyl, halo(C1-C4)alkyl, halo(C1-C4)alkoxy, -N[(C1-C4)alkyl]2, or 6-membered heterocyclyl, where the remaining variables are as described above for Formula I or any of the preceding embodiments. In an alternative, R in a compound of Formula I, II, III, or IV, or a pharma- ceutically acceptable salt thereof, 5 is F, Br, Cl, -OCH3, -OCH2CH3, OH, -O(CH2)2CH3, -NMe2, -CH(CH3)2, -C(CH3)3, -OCH(CH3)2, morpholinyl, -CH3, or -CF3, where the remaining variables are as described above for Formula I, or any of the embodiments above.

[0030] In some embodiments, the compound of formula I is selected from any of the following, or a pharma- ceutically acceptable salt thereof: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0031] The above compounds can be prepared according to the procedures described in US Pat. No. 11,091,447.

[0032] In one embodiment, the gastric cancer treated by the method is metastatic.

[0033] When used in connection with describing a chemical group that may have multiple points of attachment, a hyphen (-) indicates the point of attachment of the group to the defined variable. For example, -NH(C1-C6)alkyl means that the point of attachment of the group is on the nitrogen atom.

[0034] The terms "halo" and "halogen" refer to an atom selected from fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -B), and iodine (iodo, -I).

[0035] The term "alkyl" used alone or as part of a larger moiety such as "haloalkyl" means a saturated straight-chain or monovalent branched hydrocarbon group. Unless otherwise specified, an alkyl group typically has 1 to 4 carbon atoms (i.e., (C1-C4) alkyl).

[0036] "Alkoxy" refers to an alkyl group bonded through a linking oxygen atom and is represented by -O-alkyl. For example, "(C1-C4)alkoxy" includes methoxy, ethoxy, proproxy, and butoxy.

[0037] The term "haloalkyl" includes mono-, poly-, and perhaloalkyl groups, where the halogens are independently selected from fluorine, chlorine, bromine, and iodine.

[0038] "Haloalkoxy" is a haloalkyl group that is attached to another moiety through an oxygen atom such as, but not limited to, -OCHCF2 or -OCF3.

[0039] "Oxo" refers to the divalent functional group =O, an oxygen atom attached to another atom (typically carbon or sulfur) by a double bond.

[0040] The term "heteroaryl" refers to an aromatic ring of a particular size (e.g., 5-, 6-, 7-, 8-, or 9-membered ring) containing 1 to 4 heteroatoms independently selected from N, O, and S. Heteroaryl groups may be monocyclic or bicyclic. Monocyclic heteroaryls include, for example, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, and the like. Bicyclic heteroaryls include groups in which a monocyclic heteroaryl ring is fused to one or more aryl or heteroaryl rings. Non-limiting examples include indolyl, imidazopyridinyl, benzoxazolyl, benzoxodiazolyl, indazolyl, benzimidazolyl, benzothiazolinyl, pyrazolopyridinyl, thienopyridinyl, thienopyrimidinyl, indolizinyl, etc. When specified, any substituent on a heteroaryl group may be present at any substitutable position.

[0041] The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic ring of a particular size (e.g., 3-, 4-, 5-, 6-, or 7-membered ring) containing 1 to 4 heteroatoms independently selected from N, O, and S. Any heteroatom or carbon atom that results in a stable structure can be attached to the side groups of the heterocyclyl ring. Examples of such saturated or partially unsaturated heterocyclic groups include, but are not limited to, oxiranyl, thiiranyl, aziridinyl, tetrahydrofuranyl, tetrahydrothienyl, terahydropyranyl, pyrrolidinyl, pyridinonyl, pyrrolidonyl, piperidinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, morpholinyl, dihydrofuranyl, dihydropyranyl, dihydropyridinyl, tetrahydropyridinyl, dihydropyrimidinyl, oxetanyl, azetidinyl, and tetrahydropyrimidinyl. When specified, optional substituents on a heterocyclyl group may be present at any substitutable position, including, for example, the position at which the heterocyclyl is attached.

[0042] The term "cycloalkyl" refers to a monocyclic hydrocarbon of a particular size (e.g., 3-, 4-, 5-, 6-, or 7-membered ring). Cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, and cyclooctyl. When specified, any substituent on a cycloalkyl group may be present at any substitutable position, including, for example, the position at which the cycloalkyl is attached.

[0043] The disclosed compounds exist in various tautomeric forms and are part of this disclosure. The term "tautomer" or "tautomerism" refers to two or more interconvertible compounds / substituents resulting from the formal migration of at least one hydrogen atom and a change in at least one valence. Exemplary tautomerizations include, for example: [ka] All such isomeric forms of such compounds are expressly included. Thus, when a compound herein is represented by a structural formula or designated by a chemical name herein, all other tautomeric forms that may exist for that compound are encompassed by the structural formula. This includes compounds of formula I where X is N or C.

[0044] The compounds described herein may exist in the form of pharmaceutically acceptable salts. For use in medicine, the salts of the compounds described herein refer to non-toxic "pharmaceutically acceptable salts". The pharmaceutically acceptable salt forms include pharmaceutically acceptable acidic / anionic or basic / cationic salts. Suitable pharmaceutically acceptable acid addition salts of the compounds described herein include, for example, salts of inorganic acids (such as hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, and sulfuric acid) and salts of organic acids (such as acetic acid, benzenesulfonic acid, benzoic acid, methanesulfonic acid, and p-toluenesulfonic acid). Compounds of the present teachings having an acidic group, such as a carboxylic acid, can form pharmaceutically acceptable salts with pharmaceutically acceptable base(s). Suitable pharmaceutically acceptable base salts include, for example, ammonium salts, alkali metal salts (such as sodium and potassium salts), and alkaline earth metal salts (such as magnesium and calcium salts). Compounds with a quaternary ammonium group also contain a counterion such as chloride, bromide, iodide, acetate, perchlorate, etc. Other examples of such salts include hydrochlorides, hydrobromides, sulfates, methanesulfonates, nitrates, benzoates, and salts with amino acids such as glutamic acid.

[0045] The terms "subject" and "patient" may be used interchangeably and refer to a mammal in need of treatment, such as companion animals (e.g., dogs, cats, etc.), farm animals (e.g., cows, pigs, horses, sheep, goats, etc.), and laboratory animals (e.g., rats, mice, guinea pigs, etc.). Typically, the subject is a human in need of treatment.

[0046] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying the onset, or inhibiting the progression of gastric cancer or one or more symptoms thereof, as described herein. In some embodiments, treatment may be administered after one or more symptoms have developed, i.e., therapeutic treatment. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to susceptible individuals before symptoms develop (e.g., taking into account a history of symptoms and / or exposure to a particular microorganism or other susceptibility factor) (i.e., prophylactic treatment). Treatment may also be continued after symptoms have disappeared, e.g., to delay their recurrence.

[0047] The term "pharmaceutical acceptable carrier" refers to a non-toxic carrier, adjuvant, or vehicle that does not impair the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles that can be used in the compositions described herein include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphoric acid, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol, and wool fat.

[0048] The term "effective amount" or "therapeutically effective amount" refers to an amount of 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide that elicits a biological or medical response in a subject, e.g., a dosage of 0.01 to 100 mg / kg of body weight / day.

[0049] The compositions described herein may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, bucally, vaginally, or via an implanted reservoir. The term "parenteral" as used herein includes subcutaneous, intravenous, intramuscular, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In some embodiments, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the compositions described herein may be aqueous or oily suspensions. These suspensions may be formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents.

[0050] The specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, as well as the judgment of the treating physician, and the severity of the particular disease being treated. EXAMPLES

[0051] Gastric cancer xenograft model The Hs746T cell line is a gastric cancer epithelial cell line derived from the stomach, isolated from a metastatic site in the left leg muscle. The Hs746T cell line was implanted subcutaneously in nude mice. Tumors were allowed to grow. The mean tumor volume was 120 mm 3At the time of reaching 10 mg / kg, animals were randomly divided into four groups: vehicle control, and 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide treatment groups at 75 mg / kg, 100 mg / kg, and 150 mg / kg. Methocel E3 Premium LV with 2% Labrasol in Milli-Q water was used for the vehicle control. 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide was prepared in 2% Labrasol in Milli-Q water prior to each dose. 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide was administered by gavage twice daily (BID). In this study, 10 animals were used in each group. All animals survived throughout the 15-day experimental period. The mean tumor volume of each group over time and the tumor growth in each individual animal are plotted below. For each individual animal in the vehicle group, the TGI% was calculated based on the tumor growth inhibition of the individual tumor on each given experimental day, compared to the mean of the entire group on that given experimental day. For each group treated with 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide, the P value on each given experimental day was calculated by comparing the TGI on the same day with that of the vehicle group. This P value differs from the P value related to tumor volume instead of TGI% as specified in the CRO report. P values ​​were calculated using two-tailed distribution and two-sample equal variance (homogeneous variance) with an Excel program. ns is P>0.05, * is P<0.05, ** is P<0.01, *** is P<0.001, **** is P<0.0001.

[0052] Statistical analysis using a linear mixed effects model of tumor volume at the end of the experiment showed tumor growth inhibition of 60.3%, 71.2%, and 85.3% for the three doses, respectively. There was no significant effect on body weight. See Figure 1 and Table 1 below. These results correlate well with the dose-dependent increase in cyclin B1 and phosphohistone H3 in both tumor tissues in response to 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide. See Figure 2.

[0053] [Table 1]

[0054] SNU-5 was derived from the ascites of a patient with poorly differentiated gastric cancer who had previously received chemotherapy including 5-fluorouracil, doxorubicin, and mitomycin C, and the cell line was derived from the ascites, the site of metastasis.

[0055] The SNU-5 cell line was implanted subcutaneously in nude mice. Tumors were allowed to grow. The mean tumor volume was 122 mm. 3At the time of reaching 10 mg / kg, the animals were randomly divided into three groups: 12 animals per group, a vehicle control group, and groups treated with 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide at 75 mg / kg and 150 mg / kg. Methocel E3 Premium LV in Milli-Q water was used as the vehicle control. A solid dispersion of 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide in Milli-Q water was prepared before each dose. 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide was administered by oral gavage twice daily (BID). Twelve animals were used in each group in this study. All animals survived throughout the 23-day experimental period, except for two animals in group 2 (75 mg / kg). All animals that died before day 23 and were removed from the study were excluded from the calculations. Tumor growth inhibition percentage (TGI%) was calculated for each tumor. The mean tumor volume over time for each group and tumor growth for each individual animal are plotted below. For each individual animal in the vehicle group, the TGI% was calculated based on the tumor growth inhibition of the individual tumor on each given experimental day, compared to the mean for the entire group on that given experimental day. For each group treated with 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide, a P value was calculated for each given experimental day by comparing the TGI on that day to that of the vehicle group. This P value is different from the P value specified in the CRO report, which is related to tumor volume and not TGI%. P values ​​were calculated using two-tailed distributions and two-sample equal variances (homogeneous distributions) with the Excel program. ns: P>0.05, *: P<0.05, **: P<0.01, ***: P<0.001, ****: P<0.0001.

[0056] Results showed 64% tumor growth inhibition at 150 mg / kg. There was no significant effect on body weight. See Figure 3 and Table 2 below. These results correlate well with the dose-dependent increase in cyclin B1 and phosphohistone H3 in tumor tissue in response to 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide. See Figure 4.

[0057] [Table 2]

[0058] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly known to one of ordinary skill in the art.

Claims

1. A pharmaceutical composition for treating stomach cancer, comprising 2-(difluoromethoxy)-N-[[5-(2-methoxyphenyl)-1H-1,2,4-triazol-3-yl]methyl]benzamide or a pharmaceutically acceptable salt thereof.

2. The pharmaceutical composition of claim 1 , wherein the gastric cancer is metastatic.

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