2-(substituted benzyl)aromatic formic acid FTO inhibitor, preparation method and application thereof

Abstract

The present invention provides 2-(substituted benzyl) aromatic formic acid FTO inhibitors, preparation methods and applications thereof. Specifically, the present invention discloses 2-(substituted benzyl) aromatic formic acid compounds represented by the following general formula (I) and pharmaceutically acceptable salts, hydrates or solvates thereof, which can be used as FTO target inhibitors for treating diseases related to the FTO target, such as obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, malignant glioblastoma and the like.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical compounds. Specifically, the present invention discloses 2-(substituted benzyl) aromatic formic acid compounds represented by the following general formula (I) and pharmaceutically acceptable salts or pharmaceutically acceptable solvates thereof, which can be used as inhibitors targeting FTO to treat diseases targeting FTO, such as obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer and other cancers. Background Art

[0002] The fat mass and obesity associated protein FTO is the first protein discovered to have both m 6 A(N 6 -methyl adenosine,N 6 Fto is a protein that regulates the metabolic process in the body and regulates the activity of RNA demethylase (methylated adenosine). Fto gene knockout can lead to severe growth retardation and multiple malformations. In addition to being directly related to obesity, FTO is also closely related to metabolic syndrome (MS), type II diabetes, Alzheimer's disease and other diseases. The latest research shows that Fto is an important oncogene in the pathogenesis of solid tumors such as breast cancer and malignant glioblastoma brain tumors, and knocking down the Fto gene or reducing the expression of FTO protein can effectively inhibit the proliferation of tumor cells such as breast cancer and malignant glioblastoma brain tumors.

[0003] In summary, there is an urgent need in this field to develop small molecule inhibitors that specifically target FTO and use them in the treatment of related diseases. Summary of the invention

[0004] The purpose of the present invention is to design and synthesize a series of inhibitors that specifically target FTO, and to achieve the purpose of treating small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, malignant glioblastoma and other cancers by inhibiting the enzymatic function of FTO or the signal transduction process mediated by FTO protein.

[0005] In the first aspect of the present invention, there is provided a compound represented by the following formula (I), and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof:

[0006]

[0007] in,

[0008] A1, A2, A3, A4 are each independently CR' or N;

[0009] M is selected from the following group: CR'2, NH, O or S; R' is selected from the following group: H, halogen atoms, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, substituted or unsubstituted C1-C6 acylamino, C2-C 12 Ester group, or substituted or unsubstituted C1-C 10 alkyl;

[0010] X has a structure as shown below: carboxyl, hydroxamic acid, substituted or unsubstituted C2-C 12 an ester group, a substituted or unsubstituted amide group, a substituted or unsubstituted 3-12 membered heterocyclic group;

[0011] Y is selected from the group consisting of: substituted or unsubstituted C6-C 12 aryl, substituted or unsubstituted 3-12 membered heterocyclic group;

[0012] R a , R b , R c , R d Each is independently selected from the following group: H, halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;

[0013] The substitution refers to the substitution of one or more hydrogen atoms on the group by a substituent selected from the group consisting of a halogen atom, a carbonyl group (=O), a carboxyl group, a hydroxyl group, an amino group, a nitro group, a cyano group, a C1-C6 alkoxy group, a C1-C6 alkylamino group, a C1-C6 alkoxycarbonyl group, a C1-C6 acylamino group, a C2-C 12 Ester group, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C2-C 10 Alkenyl, substituted or unsubstituted C2-C 10 Alkynyl, substituted or unsubstituted C6-C 10 aryl, or substituted or unsubstituted five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, 3-12-membered cycloalkyl, preferably C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy or C1-C6 alkylamino; wherein the substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6-C 10 The substituents of the aryl or five-membered or six-membered heteroaryl are selected from the following groups: halogen atoms, carbonyl (=O), hydroxyl, carboxyl, C1-C6 alkoxycarbonyl, amino, C1-C6 acylamino, nitro, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C6-C6 alkyl, 10Aryl or five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, 3-12-membered cycloalkyl group, preferably a halogen atom, C1-C6 alkoxycarbonyl, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy or phenyl, 5-6-membered heterocyclic group.

[0014] The second aspect of the present invention provides a compound as shown in the following formula (I), and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof:

[0015]

[0016] in,

[0017] X has a structure shown in the following formula: carboxyl, substituted or unsubstituted C2-C12 ester, substituted or unsubstituted amide, substituted or unsubstituted 5-9 membered heterocyclic group;

[0018] Y is selected from the following group: substituted or unsubstituted C6-C12 aryl, substituted or unsubstituted 3-12 membered heterocyclic group;

[0019] The substitution refers to that one or more hydrogen atoms on the group are replaced by a substituent selected from the following groups: halogen atoms, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6-C 10 Aryl or five-membered or six-membered heteroaryl, preferably C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy or C1-C6 alkylamino; wherein the substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6-C 10 The substituents of the aryl or five-membered or six-membered heteroaryl are selected from the following groups: halogen atoms, carbonyl (=O), hydroxyl, carboxyl, C1-C6 alkoxycarbonyl, amino, C1-C6 acylamino, nitro, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C6-C6 alkyl, 10 The aryl group or the five-membered or six-membered heteroaryl group is preferably a halogen atom, a C1-C6 alkoxycarbonyl group, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, a C1-C6 alkoxy group or a phenyl group.

[0020] In another preferred embodiment, Y is not an unsubstituted phenyl group.

[0021] In another preferred embodiment, the X has a structure as shown below:

[0022]

[0023] wherein each R0 is independently selected from the following group: hydrogen, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C6-C 10 Aryl, substituted or unsubstituted C3-C 10 Cycloalkyl (including monocyclic, polycyclic, bridged ring structure), substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, -OR", R" is selected from the following group: halogen atom, carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, C2-C 12 Ester group, substituted or unsubstituted C1-C4 alkyl-C6-C 10 Aryl, substituted or unsubstituted C1-C4 alkyl-5-9 membered heterocyclic group, or substituted or unsubstituted five-membered or six-membered heteroaryl;

[0024] Y is substituted or unsubstituted benzene, substituted or unsubstituted pyridine, substituted or unsubstituted pyrimidine, substituted or unsubstituted pyridazine, substituted or unsubstituted tetrazine, substituted or unsubstituted triazine, substituted or unsubstituted pyrrole, substituted or unsubstituted thiophene, substituted or unsubstituted furan, substituted or unsubstituted tetrazole, substituted or unsubstituted triazole, substituted or unsubstituted imidazole, substituted or unsubstituted thiazole, substituted or unsubstituted oxazole, substituted or unsubstituted pyrazole, substituted or unsubstituted isothiazole, substituted or unsubstituted isoxazole, substituted or unsubstituted oxadiazole, substituted or unsubstituted thiadiazole, substituted or unsubstituted naphthalene, substituted or unsubstituted indole, substituted or unsubstituted indazole, substituted or unsubstituted quinoline, substituted or unsubstituted isoquinoline, substituted or unsubstituted benzofuran, substituted or unsubstituted benzothiophene, substituted or unsubstituted benzimidazole, substituted or unsubstituted benzoxazole, substituted or unsubstituted benzothiazole, substituted or unsubstituted benzisothiazole, substituted or unsubstituted benzisoxazole, substituted or unsubstituted benzotriazole, substituted or unsubstituted morpholine, substituted or unsubstituted dihydropiperidine, substituted or unsubstituted thiomorpholine, substituted or unsubstituted piperidine, substituted or unsubstituted piperazine, substituted or unsubstituted tetrahydropyran, substituted or unsubstituted dihydropyran, substituted or unsubstituted pyrroline, substituted or unsubstituted tetrahydrothiophene, substituted or unsubstituted tetrahydrofuran, substituted or unsubstituted oxetane, substituted or unsubstituted thietane, substituted or unsubstituted azetidine.

[0025] In another preferred embodiment, the substituent is selected from the following group: halogen atoms, hydroxyl groups, carboxyl groups, C1-C6 alkoxycarbonyl groups, amino groups, C1-C6 acylamino groups, nitro groups, cyano groups, C1-C6 alkyl groups, halogenated C1-C6 alkyl groups, C1-C6 alkoxy groups, C1-C6 alkylamino groups, C6-C 10The aryl group or the five-membered or six-membered heteroaryl group is preferably a halogen atom, a C1-C6 alkoxycarbonyl group, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, a C1-C6 alkoxy group or a phenyl group.

[0026] In another preferred embodiment, the Y is selected from the following group:

[0027]

[0028] R, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 are independently hydrogen, carbonyl (=O), C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, C1-C6 acylamino, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6-C 10 Aryl or five-membered or six-membered heteroaryl, preferably H, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy or C1-C6 alkylamino;

[0029] wherein the substituent is selected from a halogen atom, a carbonyl group (=O), a carboxyl group, a hydroxyl group, an amino group, a nitro group, a cyano group, a C1-C6 alkoxycarbonyl group, a C1-C6 acylamino group, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, a C1-C6 alkoxy group, a C1-C6 alkylamino group, a C6-C 10 The aryl group or the five-membered or six-membered heteroaryl group is preferably a halogen atom, a C1-C6 alkoxycarbonyl group, a C1-C6 alkyl group, a halogenated C1-C6 alkyl group, a C1-C6 alkoxy group or a phenyl group.

[0030] In another preferred embodiment, R a , R b , R c , R d Each is independently selected from the following group: H, F, Cl, OH, methyl, methoxy.

[0031] In another preferred embodiment, R a and R b Each independently is H, or R c and R d Each is independently H.

[0032] In another preferred embodiment, R a and R b are the same.

[0033] In another preferred embodiment, R c and R d are the same.

[0034] In another preferred embodiment, A2 and A3 are each independently CR'.

[0035] In another preferred embodiment, the X group and the Y group are the groups corresponding to the compounds in the specific embodiments.

[0036] In another preferred embodiment, the R, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 is the group corresponding to the compound in the specific embodiment.

[0037] In another preferred embodiment, the compound of formula I is selected from compounds 1-88.

[0038] In another preferred embodiment, the pharmaceutically acceptable salt is selected from the following group: an inorganic acid salt, an organic acid salt, an inorganic base salt, or an organic base salt.

[0039] In another preferred embodiment, the pharmaceutically acceptable salt is an alkali metal salt, preferably a lithium salt, a sodium salt or a potassium salt.

[0040] In another preferred embodiment, the inorganic acid salt is selected from hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic acid salt is selected from formates, acetates, trifluoroacetates, propionates, pyruvates, glycolates, oxalates, malonates, fumarates, maleates, lactates, malates, citrates, tartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, salicylates, picrates, glutamates, salicylates, ascorbates, camphorates, and camphorsulfonates.

[0041] The second aspect of the present invention provides a use of the compound of formula I as described in the first aspect of the present invention and its pharmaceutically acceptable salt, hydrate, solvate or prodrug, for use selected from the group consisting of:

[0042] (a) preparing a drug for treating a disease associated with FTO protein activity or expression;

[0043] (b) preparing targeted inhibitors of FTO protein activity;

[0044] (c) non-therapeutic inhibition of FTO protein activity in vitro; and / or

[0045] (d) Treating diseases related to FTO activity or expression.

[0046] In another preferred embodiment, the inhibition includes inhibiting m 6 A. The activity of the demethylase FTO on a DNA substrate and / or the activity of FTO on an RNA substrate.

[0047] In another preferred embodiment, the disease is selected from the following group: obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, malignant glioblastoma and other cancers.

[0048] In another preferred embodiment, when treating a disease related to FTO activity or expression, the compound of formula I is used in combination with a second therapeutic agent.

[0049] The third aspect of the present invention provides a pharmaceutical composition, which comprises: (i) an effective amount of a compound of formula I, and a pharmaceutically acceptable salt, hydrate, solvate or prodrug thereof; and (ii) a pharmaceutically acceptable carrier.

[0050] In another preferred embodiment, the carrier is a liquid, and the concentration of the compound of formula (I) in the composition is ≤300 μM; preferably ≤200 μM; more preferably ≤100 μM; and most preferably ≤30 μM.

[0051] In another preferred embodiment, the pharmaceutical composition further comprises a second therapeutic agent.

[0052] The fourth aspect of the present invention provides a method for inhibiting the activity of FTO protein, comprising the steps of: administering to the inhibited object an inhibitory effective amount of the compound of formula I or a pharmaceutically acceptable salt thereof as described in the first aspect of the present invention, or administering to the inhibited object an inhibitory effective amount of the pharmaceutical composition as described in the third aspect of the present invention.

[0053] The fifth aspect of the present invention provides a method for preparing a compound of formula I, comprising the steps of:

[0054]

[0055] In an inert solvent, the compound of formula II is reacted with a coupling reagent to obtain a compound of formula (I).

[0056] In another preferred embodiment, the coupling reagent is YB(OH)2.

[0057] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. DETAILED DESCRIPTION

[0058] After long-term and in-depth research, the inventors found that a class of compounds as shown in formula (I) can effectively and selectively inhibit the activity of FTO protein, and its activity is significantly improved compared with existing FTO inhibitors. Based on the above findings, the inventors completed the present invention.

[0059] the term

[0060] Unless otherwise specified, in this document, the term "substituted" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the following groups: C1-C 10 Alkyl, C3~C 10 Cycloalkyl, C1~C 10 Alkoxy, halogen, hydroxyl, carboxyl (-COOH), C1~C 10 Aldehyde, C2~C 10 Acyl, C2~C 10 Ester group, amino group, phenyl group; the phenyl group includes unsubstituted phenyl group or substituted phenyl group having 1-3 substituents, the substituents are selected from: halogen, C1-C 10 Alkyl, cyano, OH, nitro, C3~C 10 Cycloalkyl, C1~C 10 Alkoxy, amino.

[0061] The term "C1-C6 alkyl" refers to a straight or branched chain alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.

[0062] The term "C1-C6 alkoxy" refers to a straight or branched alkoxy group having 1 to 6 carbon atoms, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, or the like.

[0063] The term "carbonyl C1-C6 alkyl" refers to a group of the form "-COO-a straight-chain or branched alkyl group of 1 to 6 carbon atoms", for example, carbonyl-methyl, carbonyl-ethyl, carbonyl-propyl, carbonyl-isopropyl, carbonyl-butyl, carbonyl-isobutyl, carbonyl-sec-butyl, carbonyl-tert-butyl, or a similar group.

[0064] The term "C6~C 12 The term "aryl" refers to an aromatic group having 6 to 12 carbon atoms, including monocyclic or bicyclic aromatic groups, such as phenyl, naphthyl, or the like.

[0065] The term "3-12 membered heterocyclyl" refers to a saturated or unsaturated (including aromatic) ring system substituent having a 3-12 membered ring system with one or more heteroatoms selected from O, S, N or P, such as pyridyl, thienyl, piperidinyl, or similar groups, preferably a 4-9 membered heterocyclyl.

[0066] The term "halogen" refers to F, Cl, Br and I.

[0067] In the present invention, the terms "comprising", "including" or "including" indicate that various components can be used together in the mixture or composition of the present invention. Therefore, the terms "consisting mainly of" and "consisting of" are included in the term "comprising".

[0068] In the present invention, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for use in humans and / or animals without excessive adverse side effects (such as toxicity, irritation and allergic reaction), that is, has a reasonable benefit / risk ratio.

[0069] In the present invention, the term "effective amount" refers to the amount of a therapeutic agent to treat, alleviate or prevent a target disease or condition, or to an amount that exhibits a detectable therapeutic or preventive effect. The exact effective amount for a subject depends on the subject's size and health, the nature and extent of the condition, and the therapeutic agent and / or combination of therapeutic agents selected for administration. Therefore, it is useless to specify an exact effective amount in advance. However, for a given condition, the effective amount can be determined by routine experimentation, which a clinician is able to judge.

[0070] As used herein, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on the group are replaced by a substituent selected from the group consisting of halogen, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C2-C6 acyl, unsubstituted or halogenated C1-C6 alkyl-hydroxy.

[0071] Unless otherwise specified, in the present invention, all compounds appearing are intended to include all possible optical isomers, such as single chiral compounds, or mixtures of various chiral compounds (i.e., racemates). Among all compounds of the present invention, each chiral carbon atom can be optionally in R configuration or S configuration, or a mixture of R configuration and S configuration.

[0072] As used herein, the term "compound of the present invention" refers to a compound of Formula I. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compound of Formula I.

[0073] Compounds of formula I

[0074] The present invention provides a compound represented by the following formula (I), and a pharmaceutically acceptable salt and a prodrug thereof:

[0075]

[0076] in,

[0077] A1, A2, A3, A4 are each independently CR' or N;

[0078] M is selected from the following group: CR'2, NH, O or S; R' is selected from the following group: H, halogen atoms, carbonyl (=O), carboxyl, hydroxyl, amino, nitro, cyano, C1-C6 alkoxy, C1-C6 alkylamino, C1-C6 alkoxycarbonyl, substituted or unsubstituted C1-C6 acylamino, C2-C 12 Ester group, or substituted or unsubstituted C1-C 10 alkyl;

[0079] X has a structure as shown in the following formula: a carboxyl group and its bioisostere, a hydroxamic acid group, a substituted or unsubstituted C2-C 12 an ester group, a substituted or unsubstituted amide group, a substituted or unsubstituted 3-12 membered heterocyclic group;

[0080] Y is selected from the following group: substituted or unsubstituted C6-C12 aryl, substituted or unsubstituted 3-12 membered heterocyclic group;

[0081] R a , R b , R c , R d Each is independently selected from the following group: H, halogen, -OH, CN, NO2, NH2, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy;

[0082] The substitution refers to the substitution of one or more hydrogen atoms on the group by a substituent selected from the group consisting of a halogen atom, a carbonyl group (=O), a carboxyl group, a hydroxyl group, an amino group, a nitro group, a cyano group, a C1-C6 alkoxy group, a C1-C6 alkylamino group, a C1-C6 alkoxycarbonyl group, a C1-C6 acylamino group, a C2-C 12 Ester group, substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C2-C 10 Alkenyl, substituted or unsubstituted C2-C 10 Alkynyl, substituted or unsubstituted C6-C 10 aryl, or substituted or unsubstituted five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, 3-12-membered cycloalkyl, preferably C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy or C1-C6 alkylamino; wherein the substituted or unsubstituted C1-C 10 Alkyl, substituted or unsubstituted C6-C 10 The substituents of the aryl or five-membered or six-membered heteroaryl are selected from the following groups: halogen atoms, carbonyl (=O), hydroxyl, carboxyl, C1-C6 alkoxycarbonyl, amino, C1-C6 acylamino, nitro, cyano, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy, C1-C6 alkylamino, C6-C6 alkyl, 10Aryl or five-membered or six-membered heteroaryl, 3-12-membered heterocyclic group, 3-12-membered cycloalkyl group, preferably a halogen atom, C1-C6 alkoxycarbonyl, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 alkoxy or phenyl, 5-6-membered heterocyclic group.

[0083] In the present invention, the preferred compound of formula I is selected from the following group:

[0084]

[0085]

[0086]

[0087] The pharmaceutically acceptable salt is preferably an inorganic acid salt or an organic acid salt, and the inorganic acid salt is selected from hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, and acid phosphate; the organic acid salt is selected from formates, acetates, trifluoroacetates, propionates, pyruvates, glycolates, oxalates, malonates, fumarates, maleates, lactates, malates, citrates, tartrates, methanesulfonates, ethanesulfonates, benzenesulfonates, salicylates, picrates, glutamates, salicylates, ascorbates, camphorates, and camphorsulfonates.

[0088] Pharmaceutical compositions and methods of administration

[0089] Since the compounds of the present invention have excellent inhibitory activity on FTO protein, the compounds of the present invention and their various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates, and pharmaceutical compositions containing the compounds of the present invention as the main active ingredients can be used to treat, prevent and alleviate diseases related to FTO activity or expression. According to the prior art, the compounds of the present invention can be used to treat the following diseases: obesity, metabolic syndrome (MS), type 2 diabetes (T2D), Alzheimer's disease, breast cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, malignant glioblastoma brain tumors and other cancers.

[0090] The pharmaceutical composition of the present invention comprises a safe and effective amount of the compound of the present invention or a pharmacologically acceptable salt thereof and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Usually, the pharmaceutical composition contains 1-2000 mg of the compound of the present invention per dose, and more preferably, contains 5-200 mg of the compound of the present invention per dose. Preferably, the "one dose" is a capsule or tablet.

[0091] "Pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the compounds of the present invention and with each other without significantly reducing the efficacy of the compounds. Some examples of pharmaceutically acceptable carriers include cellulose and its derivatives (such as sodium carboxymethyl cellulose, sodium ethyl cellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ), wetting agents (such as sodium lauryl sulfate), colorants, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, etc.

[0092] There is no particular limitation on the administration of the compound or pharmaceutical composition of the present invention. Representative administrations include, but are not limited to, oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

[0093] Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose, and acacia; (c) humectants, for example, glycerol; (d) disintegrants, for example, agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizers, for example, paraffin; (f) absorption accelerators, for example, quaternary ammonium compounds; (g) wetting agents, for example, cetyl alcohol and glyceryl monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the case of capsules, tablets and pills, the dosage forms may also comprise buffering agents.

[0094] Solid dosage forms such as tablets, pills, capsules, pills and granules can be prepared using coatings and shell materials, such as enteric coatings and other materials known in the art. They may contain opacifiers, and the release of the active compound or compounds in such compositions can be delayed in a certain part of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be formed into microencapsulated form with one or more of the above-mentioned excipients.

[0095] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compound, the liquid dosage form may contain an inert diluent conventionally used in the art, such as water or other solvents, solubilizers and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butylene glycol, dimethylformamide and oils, in particular cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil and sesame oil or mixtures of these substances.

[0096] Besides such inert diluents, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

[0097] Suspensions, in addition to the active compounds, may contain suspending agents such as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methanol and agar, or mixtures of these substances, and the like.

[0098] Compositions for parenteral injection may include physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents or excipients include water, ethanol, polyols and suitable mixtures thereof.

[0099] Dosage forms for topical administration of the compounds of the invention include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be required.

[0100] The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

[0101] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal (such as a human) in need of treatment, wherein the dosage during administration is a pharmaceutically effective dosage, and for a person weighing 60 kg, the daily dosage is usually 1 to 2000 mg, preferably 5 to 500 mg. Of course, the specific dosage should also take into account factors such as the route of administration and the health status of the patient, which are all within the skill range of a skilled physician.

[0102] The main advantages of the present invention include:

[0103] (1) The present invention provides a new class of demethylase FTO inhibitors, which can be used at very low concentrations (IC 50 The activity of FTO protein was inhibited at 40 μM (the value was usually lower than 30 μM).

[0104] (2) The FTO inhibitor has good safety.

[0105] (3) The FTO inhibitor is highly selective and has an inhibitory effect only on FTO demethylase, and has no inhibitory effect on other dioxygenases in the same family or superfamily.

[0106] (4) The FTO inhibitor is FTO-targeting at the cellular level.

[0107] (5) The FTO inhibitor has plasma stability.

[0108] (6) The FTO protein activity inhibitor of the present invention exhibits good pharmacokinetic effects in a mouse model.

[0109] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are intended to illustrate the present invention only and are not intended to limit the scope of the present invention. The experimental methods in the following examples where specific conditions are not specified are usually performed under conventional conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or under conditions recommended by the manufacturer. Unless otherwise indicated, percentages and parts are weight percentages and weight parts.

[0110] The experimental reagents used in the experiment were mainly purchased from China National Pharmaceutical Group, Sigma and Shanghai Biotechnology.

[0111] The compound numbers mentioned in the examples are as follows:

[0112]

[0113]

[0114]

[0115] The following is an experiment on the inhibition of FTO enzyme activity by 2-(substituted benzoyl) aromatic formic acid compounds represented by general formula (I).

[0116] Example 1

[0117] Preparation of 2-(substituted benzyl)aromatic formic acid compounds 15 and 25

[0118] The synthesis of compound 25 uses o-iodobenzoic acid and 4-bromo-2,6-dichloroaniline as starting materials, undergoes Ullman coupling under the action of anhydrous copper acetate and N-methylmorpholine, undergoes esterification, Suzuki coupling reaction, and hydrolysis to obtain compound 15, on which compound 25 is obtained by hydroxamic acidization, as shown in the figure below:

[0119]

[0120] Step 1: Dissolve 30g (120mmol, 1.2eq) of o-iodobenzoic acid, 24g (100mmol, 1.0eq) of 2,6-dichloro-4-bromoaniline, triethylamine (150mmol, 1.5eq) and 9g (5.0mmol, 0.5eq) of anhydrous copper acetate in 500mL of DMF, heat to 120°C under argon protection for 24h, and after the reaction is completed, cool to room temperature, add an equal volume of water, extract the mother liquor with 300mL×3 of DCM, wash DMF with water, spin dry the organic phase, and transition the column ratio from PE:EA=20:1 to PE:EA=1:1 to obtain 9.8g of yellow solid.

[0121] Step 2: Dissolve 3.6 g of 2-(4-bromo-2,6-dichlorophenyl)amino)benzoic acid in 200 mL of anhydrous ethanol, add 20 mL of concentrated sulfuric acid under ice-water cooling, and heat to 100 ° C for reflux reaction for 12 hours. After the reaction is completed, the reaction system is cooled to room temperature, concentrated by rotary evaporation to remove ethanol, 100 mL of water is added to the system, and saturated sodium carbonate is used to neutralize until no bubbles are generated. The organic phase is extracted with ethyl acetate 50 mL × 3, and the organic phases are combined and washed with saturated brine 10 mL × 3. The organic phase is dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated and separated by silica gel chromatography to obtain 3.1 g of ethyl 2-(4-bromo-2,6-dichlorophenyl)amino)benzoate as a white solid.

[0122] Step 3: Dissolve 3.0 g (8.0 mmol, 1.0 eq) of ethyl 2-(4-bromo-2,6-dichlorophenyl)amino)benzoate, 1.36 g (9.6 mmol, 1.2 eq) of 2,5-dimethylisoxazole-4-boronic acid, 584 mg (0.8 mmol, 0.1 eq) of catalyst Pd(dppf)Cl2, and 1.68 g (12.0 mmol, 1.5 eq) of potassium carbonate in 160 mL (4:1 / v:v) of a mixed solvent of dioxane and water, and heat to 100 °C for 24 hours. Cool to room temperature, remove 1 / 2 of the solvent by rotary evaporation, extract with ethyl acetate 50 mL × 3, combine the organic phases and wash with saturated brine 10 mL × 3. The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated and separated by silica gel chromatography (petroleum ether: ethyl acetate 20:1) to obtain 1.98 g of ethyl 2-((2,6-dichloro-4-(3,5-dimethylisoxazole)phenyl)amino)benzoate as a white solid.

[0123] Step 4: Dissolve 1.98 g (4.4 mmol, 1.0 eq) of ethyl 2-((2,6-dichloro-4-(3,5-dimethylisoxazole)phenyl)amino)benzoate in a mixed solvent of 36 mL of tetrahydrofuran and 72 mL of anhydrous ethanol. Add 880 mg (22 mmol, 5.0 eq) of sodium hydroxide in 9 mL of water slowly under ice-water cooling. Heat to 45°C and react overnight. After the reaction is completed, cool the system to room temperature, concentrate to remove the organic solvent, add 20 mL of water, place in an ice-water bath, adjust the pH to 3 with 2 M dilute hydrochloric acid, and continue to stir the resulting suspension at room temperature for 30 min. Filter by suction, rinse the solid with water, and obtain 1.6 g of 2-((2,6-dichloro-4-(3,5-dimethylisoxazole)phenyl)amino)benzoic acid as a white solid, i.e., compound 15.

[0124] Step 5: Dissolve 844 mg (2.2 mmol, 1.0 eq) of 2-(4-bromo-2,6-dichlorophenyl)amino)benzoic acid in 22 mL of dichlorothionyl, heat to 65 ° C and react for two hours. Take a sample of the reaction solution and add it to anhydrous methanol. TLC tracks the end of the reaction. Rotary evaporate to dry yellow solid acid chloride for use. Weigh 152 mg (2.2 mmol, 1.0 eq) of hydroxylamine hydrochloride and dissolve it in 33 mL of a mixture of ethyl acetate and water (2:1 / v:v), place it in an ice-water bath, add 608 mg (4.4 mmol, 2.0 eq) of anhydrous potassium carbonate, and react at room temperature for one hour. Place the freshly prepared hydroxylamine solution in an ice-water bath, slowly add 10 mL of ethyl acetate solution of acid chloride (2.2 mmol, 1.0 eq) through a constant pressure dropping funnel, and react at room temperature for two hours. Rotary evaporation and slurry to obtain 623 mg of brown solid.

[0125] Other 2-(substituted benzyl)aromatic carboxylic acid compounds with pyridine and pyrazine as the parent core were prepared by a synthetic method similar to that in Example 1 using corresponding starting materials.

[0126] NMR hydrogen spectrum data of 2-(substituted benzyl) aromatic formic acid compounds:

[0127]

[0128] 1 H NMR (300MHz, d 6 -DMSO)δ13.12(s,1H),9.52(s,1H),9.42(s,1H),7.91(d,J=7.8Hz,1H),7.73(s,2H ),7.35(t,J=7.5Hz,1H),6.80(t,J=7.4Hz,1H),6.53(s,2H),6.31(d,J=4.3Hz,2H).

[0129]

[0130] 1 H NMR(300MHz,d 6 -DMSO)δ13.15(s,1H),9.51(s,1H),8.03(s,2H),7.91(d,J=7.8Hz,1H),7.81(s,1H),7.33(d,J=7.2Hz,1H),6.92(d,J=2.2Hz,1H),6.79(t,J=7.5Hz,1H),6.30(d,J=8.3Hz,1H).

[0131]

[0132] 1 H NMR(300MHz,d 6 -DMSO)δ13.19(s,1H),9.55(s,1H),7.92(d,J=7.1Hz,3H),7.79(d,J=7.5Hz,2H),7.47(dq,J=14.0,7.0Hz,3H),7.35(t,J=7.8Hz,1H),6.80(t,J=7.6Hz,1H),6.32(d,J=8.3Hz,1H).

[0133]

[0134] 1 H NMR(300MHz,d 6 -DMSO)δ13.14(s,1H),9.55(s,1H),7.96–7.85(m,3H),7.40(s,2H),7.35(t,J=7.9Hz,1H),7.06(s,1H),6.80(t,J=7.7Hz,1H),6.30(d,J=8.4Hz,1H).

[0135]

[0136] 1 H NMR(300MHz,d 6 -DMSO)δ13.23(s,1H),9.61(s,1H),8.69(s,2H),8.10(s,2H),7.92(d,J=7.7Hz,1H),7.85(d,J=5.3Hz,2H),7.36(t,J=7.6Hz,1H),6.82(t,J=7.6Hz,1H),6.33(d,J=8.5Hz,1H).

[0137]

[0138] 1 H NMR(300MHz,d 6 -DMSO)δ13.18(s,1H),9.57(s,1H),7.92(d,J=7.7Hz,1H),7.59(s,2H),7.35(dd,J=20.3,8.8Hz,5H),6.81(t,J=7.5Hz,1H),6.34(d,J=8.4Hz,1H),2.30(s,3H).

[0139]

[0140] 1 H NMR(300MHz,d 6 -DMSO)δ13.17(s,1H),9.51(s,1H),7.91(d,J=8.1Hz,1H),7.88(s,2H),7.73(d,J=2.8Hz,1H),7.67(d,J=5.0Hz,1H),7.34(t,J=7.8Hz,1H),7.19(dd,J=5.0,3.7Hz,1H),6.80(t,J=7.4Hz,1H),6.31(d,J=8.3Hz,1H).

[0141]

[0142] 1 H NMR(300MHz,d 6 -DMSO)δ9.81(s,1H),9.24(d,J=8.3Hz,3H),8.14(s,2H),7.93(d,J=7.7Hz,1H),7.33(t,J=7.5Hz,1H),6.81(t,J=7.3Hz,1H),6.31(d,J=8.3Hz,1H).

[0143]

[0144] 1 H NMR(300MHz,d 6 -DMSO)δ13.09(s,1H),9.53(s,1H),7.91(d,J=9.0Hz,3H),7.69(d,J=8.0Hz,2H),7.33(dd,J=16.2,7.9Hz,3H),6.80(t,J=7.5Hz,1H),6.31(d,J=8.3Hz,1H),2.36(s,3H).

[0145]

[0146] 1 H NMR(300MHz,d 6 -DMSO)δ13.24(s,1H),9.57(s,1H),7.98(s,2H),7.91(d,J=8.0Hz,1H),7.81–7.69(m,2H),7.35(t,J=7.5Hz,1H),6.81(t,J=7.5Hz,1H),6.32(d,J=8.5Hz,1H).

[0147]

[0148] 1 H NMR(300MHz,d 6 -DMSO)δ13.19(s,1H),9.58(s,1H),8.41(s,1H),8.10(s,2H),8.04(d,J=8.0Hz,2H),8.00–7.85(m,3H),7.63–7.50(m,2H),7.36(t,J=7.7Hz,1H),6.81(t,J=7.6Hz,1H),6.35(d,J=8.5Hz,1H).

[0149]

[0150] 1 H NMR(300MHz,d 6 -DMSO)δ13.16(s,1H),9.50(s,1H),8.70(s,2H),7.91(d,J=11.1Hz,3H),7.34(t,J=7.7Hz,1H),6.94(s,2H),6.79(t,J=7.3Hz,1H),6.28(d,J=8.4Hz,1H).

[0151]

[0152] 1 H NMR(300MHz,d 6 -DMSO)δ13.35(s,1H),9.52(s,2H),9.48–9.34(m,1H),7.90(d,J=7.7Hz,1H),7.71(s,2H),7.33(t,J=7.7Hz,1H),6.79(t,J=7.6Hz,1H),6.42(s,1H),6.25(d,J=8.4Hz,1H),3.76(s,2H),2.73(s,2H).

[0153]

[0154] 1 H NMR(300MHz,d 6 -DMSO)δ13.18(s,1H),11.39(s,1H),9.59(s,1H),7.93(d,J=7.9Hz,1H),7.85(s,2H),7.49(s,2H),7.37(t,J=7.7Hz,1H),7.21(d,J=6.2Hz,2H),6.81(t,J=7.4Hz,1H),6.61(s,1H),6.39(d,J=8.4Hz,1H).

[0155]

[0156] 1 H NMR(400MHz,DMSO)δ13.21(s,1H),9.62(s,1H),7.92(d,J=7.7Hz,1H),7.68(s,2H),7.37(t,J=7.6Hz,1H),6.82(t,J=7.6Hz,1H),6.35(d,J=8.2Hz,1H),2.47(s,3H),2.29(s,3H).

[0157]

[0158] 1 H NMR(300MHz,d 6 -DMSO)δ13.17(s,1H),9.56(s,1H),7.95(s,2H),7.92(d,J=7.9Hz,1H),7.84(d,J=8.6Hz,2H),7.55(d,J=8.6Hz,2H),7.35(t,J=7.8Hz,1H),6.81(t,J=7.5Hz,1H),6.31(d,J=8.4Hz,1H).

[0159]

[0160] 1 H NMR(500MHz,d 6 -DMSO)δ13.35(s,1H),δ9.54(s,1H),8.24–8.14(m,1H),8.05–7.99(m,1H),7.82–7.77(m,2H),7.70–7.64(m,1H),7.24(dt,J=7.0,3.8Hz,1H),2.13–2.00(m,3H),1.82–1.65(m,3H).

[0161]

[0162] 1 H NMR(500MHz,d 6 -DMSO)δ13.17(s,1H),δ9.58(s,1H),8.09–8.04(m,1H),7.96–7.91(m,1H),7.82–7.77(m,2H),7.67–7.59(m,2H),7.59–7.55(m,1H),7.26–7.19(m,1H),3.86–3.67(m,3H).

[0163]

[0164] 1 H NMR(500MHz,d 6 -DMSO)δ13.20(s,1H),δ9.52(s,1H),8.15–8.08(m,1H),8.00(dd,J=5.3,2.2Hz,1H),7.98–7.93(m,2H),7.69–7.61(m,1H),7.28–7.19(m,1H),6.26–6.18(m,1H),2.25–2.05(m,3H),2.05–1.91(m,3H).

[0165]

[0166] 1 H NMR(500MHz,d 6 -DMSO)δ13.19(s,1H),δ9.57(s,1H),8.11–8.04(m,1H),8.03–7.97(m,1H),7.83–7.76(m,2H),7.68–7.58(m,1H),7.25–7.17(m,1H).

[0167]

[0168] 1 H NMR(500MHz,d 6 -DMSO)δ13.25(s,1H),δ9.57(s,1H),8.19–8.07(m,1H),7.99–7.92(m,1H),7.72–7.67(m,2H),7.66–7.60(m,1H),7.32–7.19(m,1H),7.13–7.04(m,1H),7.05–6.96(m,1H),5.62–5.54(m,1H).

[0169]

[0170] 1 H NMR(400MHz,d 6 -DMSO)δ9.90(s,1H),8.04(d,J=7.2Hz,1H),7.66(s,2H),7.21(t,J=7.2Hz,1H),6.91(t,J=7.2Hz,1H),6.42(d,J=7.6Hz,1H),2.47(s,3H),2.29(s,3H).

[0171]

[0172] 1 H NMR(500MHz,d 6 -DMSO)δ13.35(s,1H),δ9.54(s,1H),8.24–8.14(m,1H),8.05–7.99(m,1H),7.82–7.77(m,2H),7.70–7.64(m,1H),7.24(dt,J=7.0,3.8Hz,1H),2.13–2.00(m,3H),1.82–1.65(m,3H).

[0173]

[0174] 1 H NMR(400MHz,DMSO)δ13.28(s,1H),9.54(s,1H),7.92(dd,J=7.9,1.5Hz,1H),7.54(s,1H),7.48(dd,J=11.0,1.8Hz,1H),7.44–7.36(m,1H),6.84(t,J=7.5Hz,1H),6.53(dd,J=8.3,3.9Hz,1H),2.47(s,3H),2.29(s,3H).

[0175]

[0176] 1 H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0177]

[0178] 1 H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0179]

[0180] 1 H NMR(500MHz,Chloroform)δ8.38(s,1H),8.08(s,1H),7.92(s,1H),7.62(s,1H),7.22(s,1H),6.75–6.67(m,2H),6.20(s,2H),2.30(s,6H).

[0181]

[0182] 1 H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0183]

[0184] 1 H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0185]

[0186] 1H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0187]

[0188] 1 H NMR(400MHz,DMSO)δ13.34–12.95(m,1H),9.63(s,1H),8.63(s,2H),7.93(d,J=7.9Hz,1H),7.41(d,J=7.2Hz,4H),7.32(s,1H),7.22(s,1H),7.07(d,J=8.3Hz,1H),6.79(t,J=7.6Hz,1H),2.24(d,J=7.0Hz,7H).

[0189]

[0190] 1 H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0191]

[0192] 1 H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0193]

[0194] 1H NMR(400MHz,DMSO)δ13.32(s,1H),9.53(s,1H),8.69(s,2H),7.88(dd,J=37.0,8.4Hz,5H),7.42(t,J=7.8Hz,1H),6.86(t,J=7.7Hz,1H),6.62(d,J=7.7Hz,1H).

[0195]

[0196] 1 H NMR(400MHz,DMSO)δ13.02(s,1H),7.86(dd,J=7.7,1.7Hz,1H),7.71(s,2H),7.51–7.42(m,1H),7.18(t,J=7.1Hz,1H),6.59(d,J=7.8Hz,1H),2.46(s,3H),2.28(s,3H).

[0197]

[0198] 1 H NMR(300MHz,DMSO)δ13.34–13.01(m,1H),8.06–7.96(m,2H),0.00(s,1H),0.00(t,J=5.8,5.8Hz,1H),0.00(t,J=7.5,7.5Hz,1H),0.00(t,J=7.8,7.8Hz,1H).

[0199]

[0200] 1 H NMR(400MHz,DMSO)δ13.06(s,1H),9.87(s,1H),8.25–8.12(m,2H),7.97(s,1H),7.60(s,2H),6.82(dd,J=7.6,4.8Hz,1H),2.42(s,3H).

[0201]

[0202] 1 H NMR(400MHz,DMSO)δ10.02(s,1H),8.69(s,2H),8.21(dd,J=13.1,6.2Hz,2H),8.03(s,2H),7.86(s,2H),6.85(dd,J=7.6,4.8Hz,1H).

[0203]

[0204] 1 H NMR(400MHz,DMSO)δ13.59(s,1H),9.63(s,1H),8.39(s,1H),8.22(dt,J=6.8,1.9Hz,2H),7.85(s,2H),7.79(t,J=1.7Hz,1H),7.13(d,J=1.1Hz,1H),6.84(dd,J=7.6,4.9Hz,1H).

[0205]

[0206] 1 H NMR(400MHz,DMSO)δ11.85(s,1H),8.07(s,1H),7.93(s,1H),7.60(s,2H),2.46(s,4H),2.28(s,3H).

[0207]

[0208] 1 H NMR(400MHz,DMSO)δ9.79(s,1H),8.70(d,J=6.1Hz,2H),8.32(d,J=2.2Hz,1H),8.15(d,J=2.3Hz,1H),8.06(s,2H),7.86(d,J=6.1Hz,2H).

[0209]

[0210] 1 H NMR(400MHz,DMSO)δ9.66(s,1H),8.32(d,J=2.3Hz,1H),8.13(d,J=2.3Hz,1H),7.99(s,1H),7.63(s,2H),2.42(s,1H).

[0211]

[0212] 1 H NMR(400MHz,DMSO)δ10.36(s,1H),8.40(s,1H),8.21(s,1H),8.05(s,1H),7.86(s,2H),7.79(s,1H),7.13(s,1H).

[0213]

[0214] 1H NMR(400MHz,DMSO)δ9.87(s,1H),8.69(s,2H),8.09(s,2H),7.86(s,2H),7.64(dd,J=9.5,3.0Hz,1H),7.22(td,J=8.8,3.1Hz,1H),6.34(dd,J=9.0,4.5Hz,1H).

[0215]

[0216] 1 H NMR(400MHz,DMSO)δ13.58(s,1H),9.44(s,1H),9.27(s,2H),9.24(s,1H),8.17(s,2H),7.65(dd,J=9.4,3.1Hz,1H),7.35–7.21(m,1H),6.35(dd,J=9.2,4.5Hz,1H).

[0217]

[0218] 1 H NMR(400MHz,DMSO)δ13.60(s,1H),9.37(s,1H),9.28(s,0H),7.93(s,2H),7.89–7.78(m,2H),7.64(d,J=6.7Hz,1H),7.42–7.16(m,3H),6.39–6.23(m,1H).

[0219]

[0220] 1 H NMR(400MHz,DMSO)δ13.26(s,1H),9.33(s,1H),8.26(s,2H),7.91(s,2H),7.62(dd,J=9.5,3.2Hz,1H),7.32–7.17(m,1H),6.30(dd,J=9.2,4.5Hz,1H).

[0221]

[0222] 1 H NMR(400MHz,DMSO)δ10.75(s,1H),8.76(d,J=2.8Hz,1H),8.17(dd,J=9.3,2.8Hz,1H),7.74(s,2H),6.45(d,J=9.3Hz,1H),2.48(s,3H),2.30(s,3H).

[0223]

[0224] 1 H NMR (400MHz, DMSO) δ13.49–12.92(m,1H),10.48(s,1H),9.51(s,1H),8.35(d,J=2.6Hz,1H),7.68(s,2H),7.64(dd, J=9.0,2.6Hz,1H),7.10(d,J=15.4Hz,1H),6.64(d,J=15.4Hz,1H),6.38(d,J=9.0Hz,1H),2.47(s,3H),2.29(s,3H).

[0225] NMR hydrogen spectrum of 2-(substituted benzyl) aromatic oxime acid compounds:

[0226]

[0227] 1 H NMR (400MHz, DMSO) δ11.33(s,1H),9.52(s,1H),9.17(s,1H),7.62(s,2H),7.50(d,J=7.6Hz,1H) ,7.26(t,J=7.8Hz,1H),6.79(t,J=7.5Hz,1H),6.34(d,J=8.2Hz,1H),2.44(s,3H),2.26(s,3H).

[0228]

[0229] 1 H NMR (400MHz, DMSO) δ11.32(s,1H),9.43(s,1H),7.98(s,1H),7.66(s,2H),7.52(d,J=7.7 Hz,1H),7.26(t,J=7.8Hz,1H),6.78(t,J=7.2Hz,1H),6.31(d,J=8.2Hz,1H),2.42(s,3H).

[0230]

[0231] 1 H NMR (400MHz, DMSO) δ11.33(s,1H),9.49(s,1H),7.53(d,J=7.6Hz,1H),7.49(s,2 H),7.32–7.22(m,1H),6.79(t,J=7.5Hz,1H),6.34(d,J=8.3Hz,1H),2.26(s,6H).

[0232]

[0233] 1 H NMR (400MHz, DMSO) δ11.38(s,1H),9.47(s,1H),9.22(s,1H),7.52(d,J=9.3Hz,2H),7.44(d,J=11.4Hz,1 H),7.32(t,J=7.5Hz,1H),6.84(t,J=7.3Hz,1H),6.57(dd,J=8.1,4.7Hz,1H),2.46(s,3H),2.29(s,3H).

[0234]

[0235] 1 H NMR (400MHz, DMSO) δ11.57(s,1H),10.06(s,1H),8.86(s,2H),8.19(s,2H),8.15(s,3H),7.95(d,J=7.6Hz,1H),6.87(dd,J=7.4,5.0Hz,1H).

[0236] Synthesis steps and NMR spectrum data of 2-(substituted benzyl) aromatic amide series compounds:

[0237]

[0238] In a 10mL round-bottom flask, add acid (0.2mmoL, 1.0eq), HATU (0.3mmoL, 1.5eq) dissolved in anhydrous DMF (2mL), stir at room temperature for 5min, add DIEA (0.6mmoL, 3.0eq), amine (0.4mmoL, 2.0eq) in turn. React at room temperature overnight, add water to the reaction system, and white turbidity will be generated. Filter and wash with water to obtain the target amide product.

[0239]

[0240] 1 H NMR (400MHz, d 6 -DMSO)δ10.18(s,1H),8.12(s,1H),7.74(d,J=7.6Hz,1H),7.63(s,2H),7.51(s,1H),7.27 (t,J=7.6Hz,1H),6.79(t,J=7.6Hz,1H),6.33(d,J=8.4Hz,1H),2.45(s,3H),2.27(s,3H).

[0241]

[0242] 1 H NMR(400MHz,DMSO)δ8.72(s,1H),8.14(d,J=4.7Hz,1H),8.06(d,J=7.7Hz,1H),7.59(s,2H),6.86(dd,J=7.7,4.8Hz,1H),2.65–2.53(m,5H),2.45(s,4H),2.28(s,3H),0.98(t,J=7.1Hz,6H).

[0243]

[0244] 1 H NMR(400MHz,DMSO)δ9.85(s,1H),8.53(s,1H),7.65(d,J=9.0Hz,3H),7.28(t,J=7.8Hz,1H),6.84(t,J=7.5Hz,1H),6.36(d,J=8.2Hz,1H),3.35(d,J=10.2Hz,4H),2.58(d,J=22.3Hz,5H),2.46(s,3H),2.28(s,3H),0.99(t,J=7.1Hz,6H).

[0245]

[0246] 1 H NMR(400MHz,DMSO)δ9.72(s,1H),8.53(t,J=5.5Hz,1H),7.63(d,J=5.6Hz,3H),7.28(t,J=7.7Hz,1H),6.84(t,J=7.4Hz,1H),6.36(d,J=8.3Hz,1H),2.46(s,4H),2.28(s,4H),2.11(s,3H).

[0247]

[0248] 1H NMR (400MHz, DMSO) δ13.08(s,1H),9.71(s,1H),8.49(d,J=5.7Hz,1H),8.39(s,1H),8.09(s,1H),7.88(s,2H),7.64(d,J= 7.8Hz,1H),7.24(t,J=7.8Hz,1H),6.79(t,J=7.2Hz,1H),6.28(d,J=8.3Hz,1H),2.62–2.53(m,4H),0.99(t,J=7.1Hz,6H).

[0249]

[0250] 1 H NMR (400MHz, DMSO) δ9.88(s,1H),8.67(d,J=5.3Hz,1H),7.67(d,J=6.8Hz,1H),7.64(s,2H),6.84(t,J=7.2Hz,1H),6.36(d, J=8.1Hz,1H),3.58(t,J=4.5Hz,4H),3.35–3.27(m,2H),2.46(s,3H),2.36(t,J=7.0Hz,6H),2.28(s,3H),1.77–1.66(m,2H).

[0251]

[0252] 1 H NMR (400MHz, DMSO) δ10.14(s,1H),9.89(s,1H),9.00(s,1H),8.05(s,2H),7.81(d,J=8.1Hz,1H),7.64(d,J=7.0Hz,2H),7.3 1(t,J=8.3Hz,2H),6.87(t,J=7.5Hz,1H),6.35(d,J=8.3Hz,1H),3.67(s,2H),3.24(d,J=26.4Hz,6H),1.26(d,J=6.7Hz,6H).

[0253] Synthesis steps and NMR spectrum data of 2-(substituted benzyl) aromatic ester series compounds:

[0254]

[0255] In a 10mL round-bottom flask, add acid (0.2mmoL, 1.0eq), K2CO3 (0.3mmoL, 1.5eq) and anhydrous DMF (2mL), add the corresponding bromohydrocarbon, heat to 40°C and stir overnight. Add water to the reaction system, extract the reaction system with ethyl acetate, combine the organic phases, wash the organic phases with water and saturated brine in turn, dry the organic phase with anhydrous sodium sulfate, concentrate, and separate by silica gel column chromatography to obtain the target product ester.

[0256]

[0257] 1 H NMR (500MHz, CDCl3) δ9.49 (s, 1H), 8.05 (dd, J = 8.0, 1.5Hz, 1H), 7.37–7.33 (m, 1H), 7.32 (s, 2H), 6.86–6. 81(m,1H),6.44(d,J=7.7Hz,1H),4.43(q,J=7.1Hz,2H),2.49(s,3H),2.34(s,3H),1.46(t,J=7.1Hz,3H).

[0258]

[0259] 1 H NMR (400MHz, d 6 -DMSO)δ9.29(s,1H),7.96(d,J=6.4Hz,1H),7.68(s,2H),7.51(d,J=7.8Hz,2H),7.41-7.45( m,4H),6.84(t,J=7.6Hz,1H),6.36(d,J=8.0Hz,1H),5.40(s,2H),2.49(s,3H),2.28(s,3H).

[0260]

[0261] 1 H NMR(400MHz, CDCl3)δ9.34(s,1H),8.08(dd,J=8.1,1.5Hz,1H),7.42–7.31(m,3H),6.89–6.81(m,1 H),6.45(d,J=8.5Hz,1H),4.98(d,J=2.5Hz,2H),2.57(t,J=2.5Hz,1H),2.49(s,3H),2.35(s,3H).

[0262]

[0263] 1H NMR(400MHz, CDCl3)δ9.34(s,1H),8.08(dd,J=8.1,1.5Hz,1H),7.42–7.31(m,3H),6.89–6.81(m,1 H),6.45(d,J=8.5Hz,1H),4.98(d,J=2.5Hz,2H),2.57(t,J=2.5Hz,1H),2.49(s,3H),2.35(s,3H).

[0264]

[0265] 1 H NMR(400MHz, CDCl3)δ9.34(s,1H),8.08(dd,J=8.1,1.5Hz,1H),7.42–7.31(m,3H),6.89–6.81(m,1 H),6.45(d,J=8.5Hz,1H),4.98(d,J=2.5Hz,2H),2.57(t,J=2.5Hz,1H),2.49(s,3H),2.35(s,3H).

[0266]

[0267] 1 H NMR(400MHz, CDCl3)δ9.34(s,1H),8.08(dd,J=8.1,1.5Hz,1H),7.42–7.31(m,3H),6.89–6.81(m,1 H),6.45(d,J=8.5Hz,1H),4.98(d,J=2.5Hz,2H),2.57(t,J=2.5Hz,1H),2.49(s,3H),2.35(s,3H).

[0268]

[0269] 1 H NMR(400MHz, CDCl3)δ9.34(s,1H),8.08(dd,J=8.1,1.5Hz,1H),7.42–7.31(m,3H),6.89–6.81(m,1 H),6.45(d,J=8.5Hz,1H),4.98(d,J=2.5Hz,2H),2.57(t,J=2.5Hz,1H),2.49(s,3H),2.35(s,3H).

[0270] Biological Example 1 FTO Inhibitory Activity Assay

[0271] High-purity FTO protein was obtained by nickel column affinity chromatography.

[0272] The FTO enzyme activity inhibition reaction system was as follows: 50 mM Tris·HCl, pH 7.5, 0.3 μM FTO, 1 μM 39nt-m 6 A modified double-stranded DNA, 300μM 2OG, 280μM (NH4)2Fe(SO4)2, 2mM L-Ascorbic Acid and compounds of different concentrations were incubated at room temperature for 2h, then slowly heated at 65℃ for inactivation, and 1μM 39nt antisense DNA was added to anneal into double strands. 8ul of the reaction solution was taken, and the double-stranded substrate was digested with the methylation-sensitive enzyme DpnII. The digested samples were detected by 15% non-denaturing polyacrylamide electrophoresis, and the gel was photographed under the Gel-Red staining system, and the grayscale inhibition rate of the obtained bands was read.

[0273] The following is the inhibition rate of 2-(substituted benzoyl) aromatic carboxylic acid and its derivatives represented by general formula (I) on FTO enzyme activity at a concentration of 50 μM.

[0274] In vitro inhibition rate of compound (concentration 50 μM) on FTO:

[0275]

[0276]

[0277] IC50 values ​​of FTO inhibitors of 2-(substituted benzyl)aromatic carboxylic acid and its derivatives:

[0278]

[0279]

[0280] Compared with existing FTO inhibitory compounds, the inhibitory activity of the compounds of the present application is significantly improved, and the FTO inhibitory activity of some compounds can even reach the nmol level.

[0281] Biological Example 2

[0282] The following is a study on the cytotoxicity of 2-(substituted benzyl) aromatic carboxylic acid and its derivatives FTO inhibitors represented by general formula (I) on solid tumor human small cell lung cancer cell line (SCLC-21H), human bone marrow rhabdomyosarcoma cell line (RH30) and pancreatic cancer cell line (KP3):

[0283] Solid tumor cell lines such as SCLC-21H, RH30, and KP3 were cultured separately. Cells were seeded at a density of 5000 per well in 96-well plates. The cells were cultured until they adhered to the wall. Different compounds were added and cultured for 72 hours. 10uL of MTS solution was directly added and incubated for 4 hours. The absorbance at 490nm was detected, and the inhibition rate was calculated using the DMSO group as the control.

[0284] The following is the cytotoxicity, i.e., inhibition rate, of the 2-(substituted benzoyl) aromatic formic acid compound FTO inhibitor represented by the general formula (I) at a concentration of 50 μM and a time point of 72 h on SCLC-21H, RH30, and KP3 cells.

[0285] Summary of compound activity (cell inhibition rate at a concentration of 50 μM):

[0286]

[0287]

[0288]

[0289] Cytotoxicity IC of compounds against KP3 cell line 50 :

[0290] Compound <![CDATA[IC 50 (μM)]]> 25 2.9 31 6.2 32 5.6 33 2.2 38 <1 39 2.6 78 8.7 79 2.6 80 2.0 81 <1 82 <1 83 4.7

[0291] Cytotoxicity IC of compounds against SCLC cell lines 50 :

[0292] Compound <![CDATA[IC 50 (μM)]]> 25 2.4 31 7.2 32 2.1 33 7.1 38 4.2 39 7.3 78 5.1 80 <1 79 <1 81 9.2 82 9.9

[0293] Cytotoxicity IC of compounds against cell line RH30 50 :

[0294] Compound <![CDATA[IC 50 (μM)]]> 25 2.9 31 3.6 32 5.3 33 4.1 38 9.2 39 3.2 78 14.7 79 2.6 80 2.0 81 1.3 82 1.6 83 4.7

[0295] The results show that the compounds of the present application have a very good inhibitory effect on various solid tumor cells.

[0296] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. A compound represented by the following formula (I), and a pharmaceutically acceptable salt or hydrate thereof, It is characterized in that The compound is selected from the following group:

2. The compound according to claim 1, and its pharmaceutically acceptable salt or hydrate, characterized in that: The compound is selected from the following group:

3. The compound according to claim 1, and its pharmaceutically acceptable salt or hydrate, characterized in that: The pharmaceutically acceptable salt is selected from the following group: inorganic acid salt, organic acid salt, inorganic base salt, or organic base salt.

4. The compound according to claim 1, and its pharmaceutically acceptable salt or hydrate, characterized in that: The pharmaceutically acceptable salt is an alkali metal salt.

5. A use of the compound of formula I and its pharmaceutically acceptable salt or hydrate as claimed in claim 1, characterized in that: For use selected from the group consisting of: (a) preparing a drug for treating a disease associated with FTO protein activity or expression; (b) preparing a targeted inhibitor of FTO protein activity; and / or (c) Non-therapeutic inhibition of FTO protein activity in vitro.

6. The use according to claim 5, characterized in that The disease is selected from the group consisting of obesity, metabolic syndrome, type II diabetes, Alzheimer's disease, and cancer.

7. The use according to claim 6, characterized in that The cancer is selected from the group consisting of breast cancer, small cell lung cancer, human bone marrow rhabdomyosarcoma, pancreatic cancer, and malignant glioblastoma.

8. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises: (i) an effective amount of the compound of formula I as claimed in claim 1, and a pharmaceutically acceptable salt or hydrate thereof; and (ii) a pharmaceutically acceptable carrier.

9. The pharmaceutical composition according to claim 8, characterized in that The pharmaceutical composition also includes a second therapeutic agent.

10. A method for preparing the compound of formula I as claimed in claim 1, characterized in that: The method comprises the steps of: In an inert solvent, the compound of formula II is reacted with a coupling reagent to obtain a compound of formula (I).

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