USP14 covalent inhibitor as well as preparation method and application thereof

Abstract

The invention discloses a USP14 covalent inhibitor as well as a preparation method and application thereof, and belongs to the technical field of medicines. The invention provides a compound containing a novel heterocyclic structure, which has obvious inhibitory activity on USP14, can effectively inhibit growth of human colon cancer cells, and can be used for development of a covalent USP14 inhibitor and tumor treatment.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a heterocyclic compound and a drug having a USP14 covalent inhibition effect, and further relates to a preparation method and pharmaceutical use thereof. Background Art

[0002] Ubiquitin is a small protein consisting of 76 amino acids, which is important for the regulation of protein function in cells. Ubiquitination and deubiquitination are enzymatically mediated processes, through which ubiquitin is covalently bound to a target protein or cleaved from a target protein by a deubiquitinase (DUB), in which there are about 95 kinds of DUBs in human cells, which are divided into subfamilies based on sequence homology. The USP family is characterized by their common Cys and His boxes, which contain Cys and His residues that are crucial for their DUB activity. Ubiquitination and deubiquitination processes are related to the regulation of many cellular functions, including cell cycle progression, apoptosis, modification of cell surface receptors, regulation of DNA transcription and DNA repair. Therefore, the ubiquitin system is related to the pathogenesis of many disease states, including inflammation, viral infection, metabolic dysfunction, CNS disorders and tumorigenesis.

[0003] The proteasome is a large protein complex composed of 33 different subunits. The proteasome complex functions as a protease that partially degrades unwanted or misfolded proteins. The proteasome regulates many aspects of cellular physiology, and its malfunction causes a variety of diseases, including cancer and neurodegenerative diseases. In most cases, but not exclusively, proteasome substrates are targeted for degradation by covalent attachment of multimeric chains of ubiquitin, a small, highly conserved protein. Because longer ubiquitin chains interact more strongly with the proteasome than shorter ones, processes that frequently change the length of the ubiquitin chain can also affect the rate of substrate degradation. Certain proteasome-associated deubiquitinases and ubiquitin ligases can regulate the length of ubiquitin chains attached to substrates marked for proteasome degradation. These deubiquitinases and ligases appear to regulate proteasome activity by cleaving and extending ubiquitin chains bound to the proteasome.

[0004] The mammalian proteasome contains three major deubiquitinases: Rpn11, Uch37, and Usp14. Rpn11 removes ubiquitin from labeled substrates by severing the binding site between the ubiquitin chain and the substrate. Since Rpn11-mediated cleavage occurs after the proteolytic commitment of the substrate but before the substrate is degraded, Rpn11 helps prevent ubiquitin from being degraded along with the substrate, thereby minimizing fluctuations in cellular ubiquitin levels. In addition, since proteasomal substrates must pass through a narrow translocation channel before accessing the cryptic proteolytic sites of the proteasome, the removal of large ubiquitin chains may also promote substrate translocation. Therefore, Rpn11 promotes substrate degradation by removing ubiquitin chains en bloc at a relatively late stage in the proteasomal pathway. In contrast to Rpn11, Uch37 acts before the substrate is committed to proteasomal degradation. Uch37 degrades ubiquitin chains at the ends of substrates, and its enzymatic activity only shortens the chains rather than completely removing them. It has been proposed that chain trimming by Uch37 enhances the ability of the proteasome to discriminate between short and long polyubiquitin chains.

[0005] USP14 is one of the three deubiquitinating enzymes that bind to the proteasome (USP14, Uch37, Rpn11) and can be recruited to the 19S particles to avoid recognition and degradation by the proteasome by cleaving ubiquitin chains on protein substrates. Inhibition of USP14 will lead to the rapid accumulation of polyubiquitin chain-linked proteins, endoplasmic reticulum stress and ROS production, affecting organelle transport, thereby inhibiting microtubule transport of toxic protein aggregates and inducing cell apoptosis. However, it does not affect the proteolytic activity of the 20S proteasome. After the approval of the proteasome inhibitor bortezomib for the treatment of multiple myeloma, the ubiquitin-proteasome system has gained attention as a target for the treatment of cancer. Extended treatment with bortezomib is limited by its associated toxicity and drug resistance. However, therapeutic strategies that target specific aspects of the ubiquitin-proteasome pathway upstream of the proteasome (such as DUBs) are expected to be better tolerated. Therefore, there is a need for compounds and pharmaceutical compositions that inhibit DUBs (such as USP14) for the treatment of indications in which DUB activity is observed, including, but not limited to, cancer. Summary of the invention

[0006] The purpose of the present invention is to provide a USP14 covalent inhibitor, which can be used to treat various diseases related to USP14 deubiquitinase, such as tumors.

[0007] To achieve the above-mentioned purpose of the invention, the technical solution of the present invention is as follows:

[0008] In one aspect, the present invention provides a compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs;

[0009]

[0010] in,

[0011] X is independently selected from

[0012] R1 is selected from hydrogen, hydroxyl, amino, halogen, unsubstituted or substituted alkyl, cycloalkyl or heterocycloalkyl, and the substituent is one or more of alkyl, halogen, hydroxyl or amino;

[0013] R2 is a group containing at least one of an alkenyl group, an alkynyl group, a cyano group, an acylamino group, an ethylene oxide group, and a sulfonyl fluoride group;

[0014] R3 is selected from hydrogen, hydroxy, amino, dialkylamino, halogen, C 1 4 Alkoxy, substituted or unsubstituted alkyl, cycloalkyl, phenyl, the substituent is one or more of alkyl, halogen, hydroxyl, alkoxy, amino, nitrile, and the value of n is an integer between 1 and 6;

[0015] The substituent is selected from unsubstituted or substituted aromatic heterocycles, and the substituent is selected from one or more of alkyl, halogen, hydroxyl and amino.

[0016] Preferably, R1 is selected from hydrogen, hydroxy, amino, halogen, unsubstituted or substituted with 1 to 3 substituents C 1 4 Alkyl, C 3 6 Cycloalkyl or C 3 6 Heterocycloalkyl, wherein the substituent is one or more of methyl, ethyl, fluorine, hydroxyl or amino;

[0017] R2 is selected from

[0018] R3 is selected from hydrogen, hydroxy, amino, dimethylamino, halogen, C 1 4 Alkoxy, substituted or unsubstituted C 1 4 Alkyl, C 3 6 Cycloalkyl, phenyl, the substituent is one or more of methyl, ethyl, fluorine, chlorine, hydroxyl, methoxy, amino, nitrile, and the value of n is an integer between 1 and 4;

[0019] Selected from unsubstituted or 1-3 substituent-substituted C 4-9 Aromatic heterocycle, wherein the substituent is selected from one or more of methyl, fluorine, hydroxyl and amino.

[0020] As some examples of the present invention, preferably, X is R1 is selected from hydrogen, unsubstituted or substituted with 1 to 3 substituents C1 4 Alkyl, C 3 6 Cycloalkyl or C 3 6 Heterocycloalkyl, wherein the substituent is one or more of methyl, ethyl, fluorine, hydroxyl or amino.

[0021] As some examples of the present invention, more preferably, X is R1 is selected from hydrogen, unsubstituted or substituted with 1 to 3 substituents C 3 6 Cycloalkyl or C 3 6 Heterocycloalkyl, wherein the substituent is one or more of methyl, ethyl, fluorine, hydroxyl or amino.

[0022] As some examples of the present invention, preferably, X is R1 is selected from hydrogen, unsubstituted or substituted with 1 to 3 substituents C 3 6 Heterocycloalkyl, wherein the substituent is one or more of methyl, ethyl, fluorine, hydroxyl or amino.

[0023] As some examples of the present invention, preferably, X is R2 is selected from

[0024] As some examples of the present invention, preferably, X is R3 is selected from C 1 4 Alkoxy, substituted or unsubstituted C 1 4 Alkyl, C 36 Cycloalkyl, phenyl, the substituent is one or more of methyl, ethyl, fluorine, chlorine, hydroxyl, methoxy, amino, nitrile, and the value of n is an integer between 1-4.

[0025] As some examples of the present invention, more preferably, X is R3 is selected from C 1 4 Alkoxy, substituted or unsubstituted C 3 6 Cycloalkyl, phenyl, the substituent is one or more of methyl, ethyl, fluorine, chlorine, hydroxyl, methoxy, amino, nitrile, and the value of n is an integer between 1-4.

[0026] As some examples of the present invention, preferably, X is R3 is selected from C 1 4 Alkoxy, substituted or unsubstituted C 3 6 Cycloalkyl, phenyl, the substituent is one or more of chlorine, methoxy, nitrile, and the value of n is an integer between 1-4.

[0027] As some examples of the present invention, preferably, X is Selected from substituted or unsubstituted pyridine, pyrrole, pyrazine, pyrimidine, pyridazine, thiophene, thiazole, furan, imidazole, pyrazole, triazole, tetrazole, oxazole, isoxazole, diazole, isothiazole, thiadiazole, benzotriazole, indole, isoindole, dihydroindole, isoindole, quinoxaline, quinazoline, cinnoline, chroman, isochroman, tetrahydroquinoline, quinoline, tetrahydroisoquinoline, isoquinoline, 2,3-dihydrobenzofuran, 2,3-dihydrobenzo-1,4-diene, imidazo(2,1-b)(1,3)thiazole, benzo-1,3-dioxole, and the substituent is selected from one or more of methyl, fluorine, hydroxyl, and amino.

[0028] As some examples of the present invention, more preferably, X is Selected from substituted or unsubstituted pyridine, pyrrole, pyrazine, pyrimidine, pyridazine, thiophene, thiazole, furan, imidazole, pyrazole, triazole, tetrazole, oxadiazole, isoxadiazole, isothiazole, thiadiazole, the substituent is selected from one or more of methyl, fluorine, hydroxyl, amino, the substituent is selected from one or more of methyl, fluorine, hydroxyl, amino.

[0029] As some examples of the present invention, preferably, X is Selected from substituted or unsubstituted pyrrole, thiazole, imidazole, pyrazole, triazole, tetrazole, oxazole, isoxazole, diazole, isothiazole, thiadiazole, the substituent is selected from one or more of methyl, fluorine, hydroxyl, amino.

[0030] As some examples of the present invention, preferably, X is R1 is selected from hydrogen, hydroxy, amino, halogen.

[0031] As some examples of the present invention, more preferably, X is R1 is selected from hydrogen, hydroxy, amino, fluorine, and chlorine.

[0032] As some examples of the present invention, preferably, X is R1 is hydrogen.

[0033] As some examples of the present invention, preferably, X is R2 is selected from

[0034]

[0035] As some examples of the present invention, preferably, X is R3 is selected from hydrogen, hydroxy, amino, dimethylamino, halogen, C 1 4 The value of n in the alkoxy group is an integer between 1 and 4.

[0036] As some examples of the present invention, more preferably, X is R3 is selected from methoxy or n=1.

[0037] As some examples of the present invention, preferably, X is is selected from substituted or unsubstituted benzotriazole, indole, isoindole, dihydroindole, isoindole, quinoxaline, quinazoline, cinnoline, chroman, isochroman, tetrahydroquinoline, quinoline, tetrahydroisoquinoline, isoquinoline, 2,3-dihydrobenzofuran, 2,3-dihydrobenzo-1,4-diene, imidazo (2,1-b) (1,3) thiazole, benzo-1,3-dioxole, wherein the substituent is selected from one or more of methyl, fluorine, hydroxyl and amino. As some examples of the present invention, more preferably, X is It is selected from substituted or unsubstituted indole, isoindole, dihydroindole, isohydroindole, quinoline and isoquinoline, and the substituent is selected from one or more of methyl, fluorine, hydroxyl and amino.

[0038] As some examples of the present invention, preferably, X is Selected from substituted or unsubstituted indoles, the substituents are selected from one or more of methyl, fluorine, hydroxyl and amino.

[0039] Preferably, R1 is selected from hydrogen,

[0040] R2 is selected from

[0041] R3 is selected from

[0042] Selected from

[0043] Preferably, the compound has a structure shown in formula (II) or formula (III):

[0044]

[0045]

[0046] Among them, X is selected from R1 is selected from hydrogen,

[0047] R2 is selected from

[0048]

[0049] R3 is selected from

[0050] More preferably, and as a specific example of the present invention, the compound is selected from Table 1.

[0051] Table 1

[0052]

[0053]

[0054] More preferably, and as a specific example of the present invention, the compound is selected from Table 2.

[0055] Table 2

[0056]

[0057]

[0058] Further preferably, and as a specific example of the present invention, the compound is selected from Table 3.

[0059] Table 3

[0060]

[0061] Unless otherwise specified, the term "alkyl" as used herein includes branched and straight chain saturated aliphatic hydrocarbon groups having a specified number of carbon atoms, including all isomers. Common abbreviations for alkyl groups, such as methyl, can be represented by "Me" or CH3, ethyl, can be represented by "Et" or CH2CH3, propyl, can be represented by "Pr" or CH2CH2CH3, butyl, can be represented by "Bu" or CH2CH2CH2CH3, etc. For example, "C 1-4 "C1-C4 alkyl" refers to a straight or branched chain alkyl group having a specified number of carbon atoms, including all isomers. 1-4 Alkyl includes n-, iso-, sec- and tert-butyl, n- and iso-propyl, ethyl and methyl. 1-10 "Alkyl" and the like have similar meanings.

[0062] The term "alkoxy" refers to straight and branched chain alkyl groups having the indicated number of carbon atoms attached through an oxygen bridge.

[0063] The term "halogen" (or "halo") refers to fluorine, chlorine, bromine, and iodine (alternatively referred to as fluoro (F), chloro (Cl), bromo (Br), and iodo (I)).

[0064] The term "aryl" refers to aromatic mono- and polycyclic carbon ring systems wherein the individual carbon rings in the polycyclic system are fused or linked to each other by single bonds. Typical aryl groups include phenyl, naphthyl and biphenylene.

[0065] The term "heterocycle" refers to a ring structure composed of carbon atoms and non-carbon atoms, such as nitrogen, oxygen and sulfur. Typical heterocyclic groups include pyridine, quinoline, tropane, phenothiazine, benzodiazepine, furan, pyrazolone and pyrimidine.

[0066] The term "aromatic heterocycle" refers to a 5- or 6-membered monocyclic aromatic ring or a 7- to 12-membered bicyclic ring consisting of carbon atoms and one or more heteroatoms selected from N, O and S. Examples of aromatic heterocycles include pyridyl, pyrrolyl, pyrazinyl, pyrimidinyl, pyridazinyl, thienyl (or thiophenyl), thiazolyl, furanyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl and thiadiazolyl, benzotriazolyl, indolyl, isoindolyl, indazolyl, indolinyl, isoindolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, chromanyl, isochromanyl, tetrahydroquinolyl, quinolyl, tetrahydroisoquinolyl, isoquinolyl, 2,3-dihydrobenzofuranyl, 2,3-dihydrobenzo-1,4-dienyl, imidazo(2,1-b)(1,3)thiazole and benzo-1,3-dioxolyl.

[0067] The aryl group in the term "substituted aryl" is as defined above. When the substituent of the substituted aryl group is not specified, the substituent may be selected from the following groups, including but not limited to: halogen, C1-C 20 Alkyl, CF3, NH2, N(C1-C6 alkyl)2, NO2, oxo, CN, N3, -OH, -O(C1-C6 alkyl), C3-C 10 Cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, (C0-C6 alkyl)S(O) 0-2 -, Aryl-S(O) 0-2 -, (C0-C6 alkyl)S(O) 0-2 (C0-C6 alkyl)-, (C0-C6 alkyl)C(O)NH-, H2N-C(NH)-, -O(C1-C6 alkyl)CF3, (C0-C6 alkyl)C(O)-, (C0-C6 alkyl)OC(O)-, (C0-C6 alkyl)2NC(O)-(C0-C6 alkyl)O(C1-C6 alkyl)-, (C0-C6 alkyl)C(O) 1-2 (C0-C6 alkyl)-, (C0-C6 alkyl)OC(O)NH-, aryl, aralkyl, heteroaryl, heterocyclylalkyl, halogen-aryl, halogen-aralkyl, halogen-heterocycle, halogen-heterocyclylalkyl, cyano-aryl, cyano-aralkyl, cyano-heterocycle and cyano-heterocyclylalkyl. The term "substituted phenyl" has a similar definition.

[0068] Unless explicitly stated, all ranges listed herein are inclusive. For example, "the value of n is an integer between 0 and 2" means that n can be 0, 1 or 2.

[0069] The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound of the present invention is acidic, its corresponding salt can be easily prepared from an inorganic base or an organic base. Salts derived from such inorganic bases include salts of aluminum, ammonium, calcium, copper (copper and cuprous), iron, ferrous, lithium, magnesium, manganese (manganese and manganous), potassium, sodium, zinc, etc. Preferred are salts of ammonium, calcium, magnesium, potassium and sodium. Salts prepared from organic bases include primary, secondary and tertiary amines derived from natural and synthetic sources. Pharmaceutically acceptable organic non-toxic bases that can form salts include arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydrabamine, isopropylamine, dicyclohexylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc. When the compound of the present invention is basic, its corresponding salt can be easily prepared from inorganic or organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, and the like.

[0070] The term "solvate" refers to a complex of variable stoichiometry formed by a solute (i.e., a compound of Formula I) or a pharmaceutically acceptable salt thereof and a solvent that does not interfere with the biological activity of the solute. Examples of solvents include, but are not limited to, water, ethanol, and acetic acid. When the solvent is water, the solvate is referred to as a hydrate. Hydrates include, but are not limited to, hemihydrates, monohydrates, one and a half hydrates, dihydrates, and trihydrates.

[0071] The term "prodrug" is a functional derivative of the compounds of the invention which is readily convertible in vivo into the desired compound.

[0072] In another aspect, the present invention provides a method for preparing the above-mentioned compound, comprising at least one of the following synthetic routes:

[0073] Synthetic route 1:

[0074]

[0075] Synthetic route 2:

[0076]

[0077] Synthetic route 3:

[0078]

[0079] Synthetic route 4:

[0080]

[0081] Wherein, X, R1, and R2 have the same definitions as the corresponding products.

[0082] In another aspect, the present invention provides the use of the above-mentioned compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs in the preparation of USP14 covalent inhibitors.

[0083] In another aspect, the present invention provides the use of the above-mentioned compound or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs in the preparation of anti-tumor drugs.

[0084] Preferably, the tumor is selected from one or more of cervical squamous cell carcinoma, bile duct carcinoma, hepatocellular carcinoma, gastric adenocarcinoma, sarcomatous lung cancer, thymic carcinoma, pancreatic cancer, colorectal cancer, neuroblastoma, and multiple myeloma.

[0085] In the last aspect, the present invention provides a drug comprising a compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, and one or more pharmaceutically acceptable carriers, diluents, and excipients.

[0086] The drug can be a raw material drug, a drug composition, or a drug preparation.

[0087] In the medicine, the product comprising active ingredients and inert ingredients constituting the carrier (pharmaceutically acceptable excipient) is included, as well as any product obtained directly or indirectly by the combination, complexation or aggregation of two or more ingredients, or the decomposition of one or more ingredients, or other types of reactions or interactions of one or more ingredients. Therefore, the medicine of the present invention includes any composition prepared by mixing the compound of formula I, other active ingredients and pharmaceutically acceptable excipients.

[0088] Medicine of the present invention comprises the compound shown in formula I as active ingredient (or its pharmaceutically acceptable salt or solvate), pharmaceutically acceptable carrier and optional other therapeutic ingredients or adjuvant.Pharmaceutical composition comprises composition suitable for oral, rectal, topical and parenteral (including subcutaneous, intramuscular and intravenous) administration, although the most suitable approach in any particular case depends on the nature and severity of the disease to which the active ingredient is given.Pharmaceutical composition can be prepared by any method known to the field of pharmacy.

[0089] The active ingredient can be administered orally in a solid dosage form or a liquid dosage form, such as capsules, tablets, lozenges, lozenges, granules and powders, and the liquid dosage form is such as elixirs, syrups, emulsions, dispersions and suspensions. The active ingredient can also be administered parenterally in a sterile liquid dosage form such as a dispersion, suspension or solution. Other dosage forms that can be used to administer the active ingredient include ointments, creams, drops, transdermal patches or powders for topical administration; ophthalmic solutions or suspension forms for eye administration, i.e., eye drops; sprays or powder compositions for inhalation or intranasal administration, or creams, ointments, sprays or suppositories for rectal or vaginal administration. Gelatin capsules contain active ingredients and powdered carriers, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Similar diluents can be used to prepare compressed tablets. Both tablets and capsules can be prepared as sustained-release products to provide sustained release of drugs over several hours. Compressed tablets can be coated with sugar or film to cover any unpleasant taste and protect the tablet from air, or can be enteric coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration can include colorants and flavoring agents to increase patient acceptance. Generally speaking, water, suitable oil, saline, dextrose (glucose) aqueous solution and related sugar solution and glycols such as propylene glycol or polyethylene glycol are suitable carriers of parenteral solutions. The solution for parenteral administration preferably includes water-soluble salts of active ingredients, suitable stabilizers and buffer substances used as required. Antioxidants such as sodium bisulfite, sodium sulfite or ascorbic acid alone or in combination are suitable stabilizers. Citric acid and its salts and sodium EDTA can also be used. In addition, parenteral solutions can also include preservatives, such as benzalkonium chloride, methylparaben or propylparaben and chlorobutanol. For inhalation administration, the compound of the present invention can be conveniently delivered in aerosol form from a pressurized package or a sprayer. The compound can also be delivered in the form of a powder for preparation, and the powder composition can be sucked with the help of a powder inhaler device. The preferred delivery system for suction is a metered dose inhalation (MDI) aerosol, which can be formulated into a suspension or solution of the compound of formula I in a suitable propellant, such as a fluorocarbon or hydrocarbon. For eye administration, ophthalmic preparations can be prepared with a solution or suspension of a suitable weight percentage of the compound of formula I in a suitable ophthalmic carrier, thereby keeping the compound in contact with the surface of the eye enough time to allow the compound to penetrate into the cornea and the inner area of ​​the eye.

[0090] Useful pharmaceutical dosage forms for administering the compounds of the present invention include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injection solutions, and oral suspensions.

[0091] When the compounds of the present invention are administered stepwise or in combination with other therapeutic agents, the same dosage forms as described above may be used. When the drugs are administered in physical combination, the dosage form and route of administration should be selected based on the compatibility of the combined drugs. The compounds of the present invention may be administered as the sole active ingredient or in combination with a second active ingredient, including active ingredients known to be useful for assisting in the inhibition of USP proteins and / or anti-tumor.

[0092] The beneficial effects of the present invention are as follows:

[0093] The present invention proposes a new compound that can be used as an inhibitor of the deubiquitinating enzyme USP14 protein, has the ability to effectively inhibit the activity of USP14 protein, can effectively treat and prevent USP14 protein-related and / or tumor diseases, and provides a new idea for tumor treatment. DETAILED DESCRIPTION

[0094] The following non-limiting examples can enable those skilled in the art to more fully understand the present invention, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of protection claimed in this application, and those skilled in the art can make various changes and modifications to the invention of this application based on the disclosed content, and they should also fall within the scope of protection claimed in this application.

[0095] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels.

[0096] In the following examples, the English abbreviations involved and their corresponding Chinese meanings are shown in Table 4.

[0097] Table 4

[0098]

[0099] The compounds of the present invention can be synthesized using the following four general synthetic schemes.

[0100] General solution 1

[0101]

[0102] Reagents and conditions: a) NIS, acetonitrile, rt, 4h, b) pyrrole, K2CO3, DMF, 85℃, 6h, c) n-butyllithium, THF, -78℃, 10h, d) H2, palladium on carbon, methanol, rt, 12h, e) R1-N-BOC, HATU, DIPEA, DCM, rt, 6h, f) hydrochloric acid ethyl acetate solution, rt, 1h, g) R2-Cl, TEA, DCM, rt, 1h.

[0103] General Solution 2

[0104]

[0105] Reagents and conditions: a) thionyl chloride, DMF, DCM, rt, 1h, b) 1-(4-chlorophenyl)-2,5-dimethylpyrrole, AlCl3, DCM, rt, 8h, c) H2, palladium carbon, methanol, rt, 12h, d) R1-N-BOC, HATU, DIPEA, DCM, rt, 6h, e) hydrochloric acid ethyl acetate solution, rt, 1h, f) R2-Cl, TEA, DCM, rt, 1h.

[0106] General solution 3

[0107]

[0108] Reagents and conditions: a) thionyl chloride, DMF, DCM, rt, 1h, b) 5-methoxyindole, AlCl3, DCM, rt, 8h, c) borane dimethyl sulfide, rt, 8h, d) boron tribromide, DCM, rt, 3h, e) (2-bromomethyl)dimethylamine, KOH, ethanol, 60°C, 6h, f) H2, palladium on carbon, methanol, rt, 12h, g) acryloyl chloride, TEA, DCM, rt, 1h.

[0109] General solution 4

[0110]

[0111] Reagents and conditions: a) thionyl chloride, DMF, DCM, rt, 1 h, b) 5-methoxyindole, AlCl3, DCM, rt, 8 h, c) H2, Raney nickel, methanol, rt, 12 h, d) 4-(dimethylamino)but-2-enoic acid hydrochloride, HATU, DIPEA, DCM, rt, 6 h.

[0112] The present invention is further described below with reference to the embodiments.

[0113] Example 1 N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)acrylamide

[0114] Synthesized according to Scheme 1 steps a, b, c, d, g.

[0115]

[0116] Step 1. Dissolve methyl 4-chloro-2-nitrobenzoate (108 mg, 0.5 mmol) in N,N-dimethylformamide (2 ml), add potassium carbonate (212 mg, 1.5 mmol) and tetrahydropyrrole (42 mg, 0.6 mmol) in turn, and stir at 85°C for 2 h. After the reaction is completed, add water to the reaction solution to quench, extract with ethyl acetate 5 times, backwash with saturated brine 2 times, dry the organic phase with anhydrous sodium sulfate, and purify by column chromatography (PE:EA=7:1) to obtain 127 mg of methyl 2-nitro-4-(pyrrolidin-1-yl)benzoate. ESI-MS: m / z=251[M+H] + . Step 2. Weigh 205 mg (1 mmol) of 1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole and add it to 8 ml of acetonitrile, then add 248 mg (1.1 mmol) of N-iodosuccinimide and stir at room temperature for 3 hours. TLC detected that the reaction was complete, and the organic phase was extracted and washed with water several times with DCM, dried with MgSO4, and the crude product was separated by flash column chromatography with an elution gradient of pure DCM. The products were combined, concentrated and dried to obtain 272 mg of 1-(4-chlorophenyl)-3-iodo-2,5-dimethyl-1H-pyrrole. ESI-MS: m / z=332[M+H] + .

[0117] Step 3. Weigh 331 mg (1 mmol) of 1-(4-chlorophenyl)-3-iodo-2,5-dimethyl-1H-pyrrole and add it to anhydrous tetrahydrofuran. After cooling to -78°C under nitrogen protection, add 77 mg (1.2 mmol) of n-butyl lithium. After stirring for 1 hour at -78°C under nitrogen protection, add 250 mg (1 mmol) of methyl 2-nitro-4-(pyrrolidin-1-yl)benzoate, and then stir at room temperature overnight. TLC detection showed no change in the reaction. The organic phase was extracted and washed with water several times with DCM, dried with MgSO4, and the crude product was separated by flash column chromatography with an elution gradient of pure DCM. The products were combined, concentrated and dried to obtain 101 mg of (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-yl)(2-nitro-4-(pyrrolidin-1-yl)phenyl)methanone. ESI-MS: m / z=424[M+H] + .

[0118] Step 4. Weigh 42.3 mg (0.1 mmol) of (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) (2-nitro-4-(pyrrolidin-1-yl) phenyl) ketone and dissolve it in 4 ml of methanol, then add 4.3 mg (1%) of palladium carbon and stir at room temperature overnight. TLC detection shows that the reaction is complete. After filtration, the product (2-amino-4-(pyrrolidin-1-yl) phenyl) (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) ketone (37.2 mg) was obtained. ESI-MS: m / z=393 [M+H] + ;

[0119] Step 5. Weigh 39 mg (0.1 mmol) of (2-amino-4-(pyrrolidin-1-yl)phenyl)(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-yl)methanone and dissolve it in 4 ml of anhydrous dichloromethane mixed with 30.3 mg (0.3 mmol) of triethylamine, then add 8 mg (0.09 mmol) of acryloyl chloride and stir at room temperature for 1 hour. TLC detected that the reaction was complete, and the organic phase was extracted and washed with water several times with DCM, dried over MgSO4, and the crude product was separated by flash column chromatography with an elution gradient of MeOH:DCM=1:10. The products were combined, concentrated and dried to obtain 21 mg of N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)acrylamide. ESI-MS: m / z=448[M+H] + ; 1 H NMR(400MHz,Chloroform-d)δ12.27(s,1H),8.07(d,J=2.4Hz,1H),7.79(d,J=8.9Hz,1H),7.51-7.44(m,2H),7.21-7.12(m,2H),6.42 -6.30(m,2H),6.21(dd,J=8.9,2.5Hz,1H),6.14(s,1H),5.72(dd,J=9.7,1.8Hz,1H),3.41(d,J=6.2Hz,4H),2.17(s,3H),1.98(s,3H).

[0120] Example 2 N-(5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)acrylamide

[0121] Synthesized according to Scheme 2 steps a, b, c, f.

[0122]

[0123] Step 1. Weigh 236 mg (1 mmol) of 3-nitro-4-(pyrrolidin-1-yl)benzoic acid and add it to 10 ml of dry dichloromethane solution mixed with three drops of N,N-dimethylformamide, then add 476 mg (4 mmol) of thionyl chloride and stir at room temperature for 1 hour. TLC detection shows that the reaction is complete, and the product 3-nitro-4-(pyrrolidin-1-yl)benzoyl chloride is obtained by concentration and drying for use and directly used in the next step.

[0124] Step 2. Aluminum chloride (27 mg, 0.2 mmol) was added to anhydrous DCM (1 ml), stirred at 0°C and 3-nitro-4-(pyrrolidin-1-yl)benzoyl chloride (51 mg, 0.2 mmol) was slowly added, and stirred at 0°C for half an hour. Subsequently, 1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole (20 mg, 0.1 mmol) was dissolved in anhydrous DCM (1 ml), and chloroacetyl chloride was slowly added dropwise to the aluminum chloride DCM solution at 0°C, and stirred at room temperature overnight. After the reaction was completed, water was added to the reaction solution to quench, and the mixture was extracted with DCM (dichloromethane, 3×20 ml). The organic phase was dried over anhydrous sodium sulfate and purified by column chromatography (DCM:CH3OH=40:1) to obtain 22 mg of the product (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl)(3-nitro-4-(pyrrolidin-1-yl)phenyl)methanone. ESI-MS: m / z=424[M+H] + .

[0125] Step 3. Weigh 42.3 mg (0.1 mmol) of (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) (3-nitro-4-(pyrrolidin-1-yl) phenyl) ketone and dissolve it in 4 ml of methanol, then add an appropriate amount of Raney nickel and stir overnight at room temperature. TLC detection shows that the reaction is complete. After filtration, the product (3-amino-4-(pyrrolidin-1-yl) phenyl) (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) ketone (37.2 mg) was obtained. ESI-MS: m / z=393 [M+H] + ;

[0126] Step 4. Weigh 39 mg (0.1 mmol) of (3-amino-4-(pyrrolidin-1-yl)phenyl)(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrolidin-3-yl)methanone and dissolve it in 4 ml of anhydrous dichloromethane mixed with 30.3 mg (0.3 mmol) of triethylamine, then add 8 mg (0.09 mmol) of acryloyl chloride and stir at room temperature for 1 hour. TLC detected that the reaction was complete, and the organic phase was extracted and washed with water several times with DCM, dried over MgSO4, and the crude product was separated by flash column chromatography with an elution gradient of MeOH:DCM=1:10. The products were combined, concentrated and dried to obtain 15 mg of N-(5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)acrylamide. ESI-MS: m / z=448[M+H] + ;1H NMR(400MHz,Chloroform-d)δ8.30(s,1H),7.88(s,1H),7.68-7.59(m,1H),7.48-7.46(m,2H),7.17-7.15(m,2H),6.95(d,J=8.6Hz,1 H),6.44-6.33(m,2H),6.29(s,1H),5.78-5.70(m,1H),3.21(d,J=6.4Hz,4H),2.26(s,3H),1.98(d,J=1.0Hz,3H),1.94-1.91(m,4H).

[0127] Example 3 N-(2-((5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)acrylamide

[0128] Synthesized according to Scheme 2 steps a, b, c, d, e, f.

[0129]

[0130] Step 1. Weigh 236 mg (1 mmol) of 3-nitro-4-(pyrrolidin-1-yl)benzoic acid and add it to 10 ml of dry dichloromethane solution mixed with three drops of N,N-dimethylformamide, then add 476 mg (4 mmol) of thionyl chloride and stir at room temperature for 1 hour. TLC detection shows that the reaction is complete, and the product 3-nitro-4-(pyrrolidin-1-yl)benzoyl chloride is obtained by concentration and drying for use and directly used in the next step.

[0131] Step 2. Aluminum chloride (27 mg, 0.2 mmol) was added to anhydrous DCM (1 ml), stirred at 0°C and 3-nitro-4-(pyrrolidin-1-yl)benzoyl chloride (51 mg, 0.2 mmol) was slowly added, and stirred at 0°C for half an hour. Subsequently, 1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole (20 mg, 0.1 mmol) was dissolved in anhydrous DCM (1 ml), and chloroacetyl chloride was slowly added dropwise to the aluminum chloride DCM solution at 0°C, and stirred at room temperature overnight. After the reaction was completed, water was added to the reaction solution to quench, and the mixture was extracted with DCM (dichloromethane, 3×20 ml). The organic phase was dried over anhydrous sodium sulfate and purified by column chromatography (DCM:CH3OH=40:1) to obtain 22 mg of the product (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl)(3-nitro-4-(pyrrolidin-1-yl)phenyl)methanone. ESI-MS: m / z=424[M+H] + .

[0132] Step 3. Weigh 42.3 mg (0.1 mmol) of (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) (3-nitro-4-(pyrrolidin-1-yl) phenyl) ketone and dissolve it in 4 ml of methanol, then add an appropriate amount of Raney nickel and stir overnight at room temperature. TLC detection shows that the reaction is complete. After filtration, the product (3-amino-4-(pyrrolidin-1-yl) phenyl) (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) ketone (37.2 mg) was obtained. ESI-MS: m / z=393 [M+H] + ;

[0133] Step 4. Weigh 17.5 mg (0.1 mmol) of (tert-butyloxycarbonyl)glycine and add it to 2 ml of dichloromethane, then add 57 mg (0.15 mmol) of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and 39 mg (0.3 mmol) of N,N-diisopropylethylamine in sequence, stir at room temperature for 1 hour, then add 39 mg (0.1 mmol) of (3-amino-4-(pyrrolidin-1-yl)phenyl)(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl)methanone, stir at room temperature for 4 hours. TLC detection shows that the reaction is complete, and the organic phase is extracted and washed with water several times with DCM, dried with MgSO4, and the crude product is separated by flash column chromatography with an elution gradient of EA:PE=1:5. The products were combined, concentrated and dried to obtain 22 mg of tert-butyl (2-((5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)carbamate. ESI-MS: m / z=551[M+H] + ;

[0134] Step 5. Weigh 55 mg (0.1 mmol) of tert-butyl (2-((5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)carbamate and add it to 0.5 ml (1 mmol, 2M) hydrochloric acid ethyl acetate solution and stir at room temperature for 0.5 hours. TLC detected that the reaction was complete, and the reaction was quenched with saturated sodium bicarbonate aqueous solution. The organic phase was extracted and washed with water several times with EA, dried over MgSO4, and the products were combined, concentrated and dried to obtain 42 mg of 2-amino-N-(5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)acetamide. ESI-MS: m / z=451[M+H] + ;

[0135] Step 6. Weigh 45 mg (0.1 mmol) of 2-amino-N-(5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)acetamide and dissolve it in 4 ml of anhydrous dichloromethane mixed with 30.3 mg (0.3 mmol) of triethylamine, then add 8 mg (0.09 mmol) of acryloyl chloride and stir at room temperature for 1 hour. TLC detection shows that the reaction is complete, and the organic phase is extracted and washed with water several times with DCM, dried over MgSO4, and the crude product is separated by flash column chromatography with an elution gradient of MeOH:DCM=1:10. The products are combined, concentrated and dried to obtain 19 mg of N-(2-((5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)acrylamide. ESI-MS: m / z = 505 [M + H] + ; 1 HNMR(400MHz,Chloroform-d)δ8.27(s,1H),8.19(d,J=2.0Hz,1H),7.64(dd,J=8.5,2 .1Hz,1H),7.49-7.46(m,2H),7.17(d,J=6.6Hz,2H),6.93(d,J=8.5Hz,1H),6.36(dd,J =16.9,1.6Hz,1H),6.28-6.24(m,2H),5.71(dd,J=10.1,1.6Hz,1H),4.18(d,J=5.4Hz, 2H), 3.20 (t, J = 5.7Hz, 4H), 2.27 (s, 3H), 1.98 (d, J = 0.9Hz, 3H), 1.92 (q, J = 3.3Hz, 4H).

[0136] Example 4 N-(3-((5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)amino)-3-oxopropyl)acrylamide

[0137]

[0138] Synthesized using a similar method as described in Example 3, using BOC-β-alanine in step d.

[0139] ESI-MS: m / z = 519 [M + H] + ; 1 H NMR(400MHz,Chloroform-d)δ8.14(d,J=2.2Hz,1H),7.85(s,1H),7.64(dd,J=8.5,2.1Hz,1H ),7.50-7.46(m,2H),7.18-7.15(m,2H),6.91(d,J=8.5Hz,1H),6.28-6.25(m,1H),6.23(d,J= 1.6Hz,1H),6.07(dd,J=17.0,10.2Hz,1H),5.60(dd,J=10.2,1.6Hz,1H),3.68(q,J=5.9Hz,2H ), 3.26-3.19 (m, 4H), 2.68 (t, J = 5.8Hz, 2H), 2.26 (s, 3H), 1.98 (s, 3H), 1.92 (q, J = 3.4Hz, 4H).

[0140] Example 5 N-(5-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)-3-propionamidopropionamide

[0141]

[0142] Synthesized using a similar method as described in Example 4 using propionyl chloride in step f.

[0143] ESI-MS: m / z = 521 [M + H] +;1H NMR(400MHz,Chloroform-d)δ8.15(d,J=2.1Hz,1H),7.83(s,1H),7.64(dd,J=8.5 ,2.1Hz,1H),7.49-7.46(m,2H),7.18-7.15(m,2H),6.92(d,J=8.5Hz,1H),6.25(s, 1H),3.60(q,J=5.9Hz,2H),3.26-3.19(m,4H),2.64(t,J=5.8Hz,2H),2.26(s,3H) ,2.17(q,J=7.6Hz,2H),1.98(s,3H),1.93(q,J=3.3Hz,4H),1.11(t,J=7.6Hz,3H).

[0144] Example 6 N-(2-((2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)acrylamide

[0145] Synthesized according to Scheme 1 steps a, b, c, d, e, f, g.

[0146]

[0147] Step 1. Dissolve methyl 4-chloro-2-nitrobenzoate (108 mg, 0.5 mmol) in N,N-dimethylformamide (2 ml), add potassium carbonate (212 mg, 1.5 mmol) and tetrahydropyrrole (42 mg, 0.6 mmol) in turn, and stir at 85°C for 2 h. After the reaction is completed, add water to the reaction solution to quench, extract with ethyl acetate 5 times, backwash with saturated brine 2 times, dry the organic phase with anhydrous sodium sulfate, and purify by column chromatography (PE:EA=7:1) to obtain 127 mg of methyl 2-nitro-4-(pyrrolidin-1-yl)benzoate. ESI-MS: m / z=251[M+H] + . Step 2. Weigh 205 mg (1 mmol) of 1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole and add it to 8 ml of acetonitrile, then add 248 mg (1.1 mmol) of N-iodosuccinimide and stir at room temperature for 3 hours. TLC detected that the reaction was complete, and the organic phase was extracted and washed with water several times with DCM, dried with MgSO4, and the crude product was separated by flash column chromatography with an elution gradient of pure DCM. The products were combined, concentrated and dried to obtain 272 mg of 1-(4-chlorophenyl)-3-iodo-2,5-dimethyl-1H-pyrrole. ESI-MS: m / z=332[M+H] + .

[0148] Step 3. Weigh 331 mg (1 mmol) of 1-(4-chlorophenyl)-3-iodo-2,5-dimethyl-1H-pyrrole and add it to anhydrous tetrahydrofuran. After cooling to -78°C under nitrogen protection, add 77 mg (1.2 mmol) of n-butyl lithium. After stirring for 1 hour at -78°C under nitrogen protection, add 250 mg (1 mmol) of methyl 2-nitro-4-(pyrrolidin-1-yl)benzoate, and then stir at room temperature overnight. TLC detection showed no change in the reaction. The organic phase was extracted and washed with water several times with DCM, dried with MgSO4, and the crude product was separated by flash column chromatography with an elution gradient of pure DCM. The products were combined, concentrated and dried to obtain 101 mg of (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-yl)(2-nitro-4-(pyrrolidin-1-yl)phenyl)methanone. ESI-MS: m / z=424[M+H] + .

[0149] Step 4. Weigh 42.3 mg (0.1 mmol) of (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) (2-nitro-4-(pyrrolidin-1-yl) phenyl) ketone and dissolve it in 4 ml of methanol, then add 4.3 mg (1%) of palladium carbon and stir at room temperature overnight. TLC detection shows that the reaction is complete. After filtration, the product (2-amino-4-(pyrrolidin-1-yl) phenyl) (1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl) ketone (37.2 mg) was obtained. ESI-MS: m / z=393 [M+H] + ;

[0150] Step 5. Weigh 17.5 mg (0.1 mmol) of (tert-butyloxycarbonyl)glycine and add it to 2 ml of dichloromethane, then add 57 mg (0.15 mmol) of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and 39 mg (0.3 mmol) of N,N-diisopropylethylamine in sequence, stir at room temperature for 1 hour, then add 39 mg (0.1 mmol) of (2-amino-4-(pyrrolidin-1-yl)phenyl)(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrol-3-yl)methanone, stir at room temperature for 4 hours. TLC detection shows that the reaction is complete, and the organic phase is extracted and washed with water several times with DCM, dried with MgSO4, and the crude product is separated by flash column chromatography with an elution gradient of EA:PE=1:5. The products were combined, concentrated and dried to obtain 22 mg of tert-butyl (2-((2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)carbamate. ESI-MS: m / z=551[M+H] + ;

[0151] Step 6. Weigh 55 mg (0.1 mmol) of tert-butyl (2-((2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)carbamate and add it to 0.5 ml (1 mmol, 2M) hydrochloric acid ethyl acetate solution and stir at room temperature for 0.5 hours. TLC detected that the reaction was complete, and the reaction was quenched with saturated sodium bicarbonate aqueous solution. The organic phase was extracted and washed with water several times with EA, dried over MgSO4, and the products were combined, concentrated and dried to obtain 42 mg of 2-amino-N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)acetamide. ESI-MS: m / z=451[M+H] + ;

[0152] Step 7. Weigh 45 mg (0.1 mmol) of 2-amino-N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-2-(pyrrolidin-1-yl)phenyl)acetamide and dissolve it in 4 ml of anhydrous dichloromethane mixed with 30.3 mg (0.3 mmol) of triethylamine, then add 8 mg (0.09 mmol) of acryloyl chloride and stir at room temperature for 1 hour. TLC detection shows that the reaction is complete, and the organic phase is extracted and washed with water several times with DCM, dried over MgSO4, and the crude product is separated by flash column chromatography with an elution gradient of MeOH:DCM=1:10. The products are combined, concentrated and dried to obtain 17 mg of N-(2-((2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)acrylamide. ESI-MS: m / z = 506 [M + H] + ; 1 HNMR(400MHz,Chloroform-d)δ7.86(d,J=2.4Hz,1H),7.79(d,J=8.9Hz,1H),7.52-7.4 5(m,2H),7.21-7.15(m,2H),6.70(s,1H),6.34(d,J=1.7Hz,1H),6.28(d,J=10.0Hz,1H ),6.25-6.21(m,1H),6.13(d,J=1.2Hz,1H),5.68(ddd,J=10.1,1.7,0.8Hz,1H),4.28- 4.22(m,2H),3.44-3.36(m,4H),2.17(s,3H),2.06-2.01(m,4H),1.99(d,J=0.9Hz,3H).

[0153] Example 7 2-Chloro-N-(2-((2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)amino)-2-oxoethyl)acetamide

[0154]

[0155] Synthesized using a similar method as described in Example 6 using chloroacetyl chloride in step g.

[0156] ESI-MS: m / z = 527 [M + H] + ; 1 H NMR(400MHz,Chloroform-d)δ7.87(d,J=2.4Hz,1H),7.80(d,J=8.9Hz,1H),7.51-7.45(m,2H),7.36(t,J=5.3Hz,1H),7.20-7.15(m,2H), 6.13 (d, J = 1.0Hz, 1H), 4.19 (d, J = 5.9Hz, 4H), 3.40 (td, J = 6.7, 5.4, 3.2Hz, 4H), 2.18 (s, 3H), 2.01 (d, J = 3.3Hz, 4H), 1.98 (d, J = 1.0Hz, 3H).

[0157] Example 8 N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)-2-cyanaminoacetamide

[0158]

[0159] Synthesized using a similar method as described in Example 6 using cyanogen bromide in step g.

[0160] ESI-MS: m / z = 476 [M + H] + ; 1 H NMR(400MHz,Chloroform-d)δ7.83-7.78(m,2H),7.50(d,J=2.8Hz,1H),7.49-7.48(m,1H),7.47(d,J=2.7Hz,1H),7.18(d,J=2.1Hz,1H),7.16 (d,J=2.0Hz,1H),6.12(d,J=1.1Hz,1H),3.98(d,J=5.1Hz,2H),3.40(d,J=4.6Hz,4H),2.17(s,3H),2.03-2.01(m,4H),1.98(d,J=0.9Hz,3H).

[0161] Example 9 N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)-2-(prop-2-yn-1-ylamino)acetamide

[0162]

[0163] Synthesized using a similar method as described in Example 6 using 3-bromopropyne in step g.

[0164] ESI-MS: m / z = 489 [M + H] + ; 1 H NMR(400MHz,Chloroform-d)δ7.94(d,J=2.4Hz,1H),7.75(d,J=8.8Hz,1H),7.50-7.44(m,2H),7.19-7.14(m,2H),6.21(dd,J=8.9,2.5Hz, 1H), 6.14 (d, J = 1.1Hz, 1H), 3.65 (s, 2H), 3.62 (d, J = 2.5Hz, 2H), 3.44-3.37 (m, 4H), 2.26 (t, J = 2.4Hz, 1H), 2.04-2.00 (m, 5H), 1.98 (s, 3H).

[0165] Example 10 N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)-2-(di(prop-2-yn-1-yl)amino)acetamide

[0166]

[0167] Synthesized using a similar method as described in Example 9.

[0168] ESI-MS: m / z = 527 [M + H] + ;1H NMR(400MHz,Chloroform-d)δ7.94(d,J=2.4Hz,1H),7.74(d,J=8.9Hz,1H),7. 52-7.41(m,2H),7.22-7.13(m,2H),6.20(dd,J=8.9,2.5Hz,1H),6.15(t,J=1. 0Hz,1H),3.62(d,J=2.4Hz,4H),3.45(s,2H),3.39(td,J=5.4,4.2,2.5Hz,4H) ,2.23(t,J=2.4Hz,2H),2.16(s,3H),2.05-1.99(m,4H),1.98(d,J=1.0Hz,3H).

[0169] Example 11 N-(3-(5-(2-(dimethylamino)ethoxy)-1H-indole-3-carbonyl)phenyl)acrylamide was synthesized according to Scheme 3 steps a, b, c, e, f, g.

[0170]

[0171] Step 1. Dissolve 167 mg (1 mmol) of 3-nitrobenzoic acid in 3 ml of anhydrous dichloromethane, add 3 drops of N,N-dimethylformamide, then add 357 mg (3 mmol) of thionyl chloride, and stir at room temperature for 30 minutes. TLC detection shows that the reaction is complete, and the mixture is concentrated under reduced pressure to obtain a light yellow solid 3-nitrobenzoyl chloride for use.

[0172] Step 2. Dissolve 3-nitrobenzoyl chloride in 3 ml of anhydrous dichloromethane, then add 133 mg (1 mmol) of aluminum chloride, and stir at 0°C for 30 minutes under nitrogen protection. After the aluminum chloride solid is dissolved, add 147 mg (1 mmol) of 5-methoxyindole at 0°C, then return to room temperature and stir for 6 hours under nitrogen protection. TLC detection reaction is complete, use DCM to extract and wash the organic phase with water for many times, dry MgSO4, and separate the crude product by flash column chromatography, with an elution gradient of EA: PE = 1: 2. The products are combined, concentrated and dried to obtain 212 mg of (5-methoxyindol-3-yl) (3-nitrophenyl) ketone. ESI-MS: m / z = 297 [M+H] + ; Step 3. Weigh 296 mg (1 mmol) (5-methoxyindol-3-yl) (3-nitrophenyl) ketone and dissolve it in 4 ml of anhydrous dichloromethane, then add 250 mg (1 mmol) of boron tribromide and stir at room temperature for 2 hours. TLC detection reaction is complete, DCM is used to extract and wash the organic phase with water for multiple times, and MgSO4 is dried. The crude product is separated by flash column chromatography with an elution gradient of MeOH: DCM = 1: 20. The products are combined, concentrated and dried to obtain 101 mg of (5-hydroxy-1H-indol-3-yl) (3-nitrobenzene) ketone. ESI-MS: m / z = 283 [M + H] + ;

[0173] Step 4. Weigh 282 mg (1 mmol) of (5-hydroxy-1H-indol-3-yl) (3-nitrobenzene) ketone and add it to 6 ml of ethanol containing 112 mg (2 mmol) of potassium hydroxide. Heat to 60°C and stir for 5 minutes. Then add 233 mg (1 mmol) of N, N-dimethylaminoethyl bromide hydrobromide and stir at 60°C for 4 hours. TLC detection shows that the reaction is complete. DCM is used to extract and wash the organic phase with water for multiple times, and then dried over MgSO4. The crude product is separated by flash column chromatography with an elution gradient of MeOH:DCM=1:20. The products are combined, concentrated and dried to obtain 177 mg of (5-(2-(dimethylamino)ethoxy)-1H-indol-3-yl) (3-nitrophenyl) ketone. ESI-MS: m / z=354[M+H] + ;

[0174] Step 5. Weigh 353 mg (1 mmol) of (5-(2-(dimethylamino)ethoxy)-1H-indol-3-yl) (3-nitrophenyl)methanone and dissolve it in 4 ml of methanol, then add 3.4 mg (1%) of palladium carbon and stir at room temperature overnight. TLC detected that the reaction was complete, filtered, concentrated and dried to obtain 344 mg of the product (3-aminophenyl) (5-(2-(dimethylamino)ethoxy)-1H-indol-3-yl)methanone. ESI-MS: m / z=324 [M+H] + ;

[0175] Step 6. Weigh 323 mg (1 mmol) of (3-aminophenyl)(5-(2-(dimethylamino)ethoxy)-1H-indol-3-yl)methanone and dissolve it in 4 ml of anhydrous dichloromethane mixed with 303 mg (3 mmol) of triethylamine, then add 81 mg (0.9 mmol) of acryloyl chloride and stir at room temperature for 1 hour. TLC detection shows that the reaction is complete, and the organic phase is extracted and washed with water several times with DCM, dried over MgSO4, and the crude product is separated by flash column chromatography with an elution gradient of MeOH:DCM=1:10. The products are combined, concentrated and dried to obtain 67 mg of N-(3-(5-(2-(dimethylamino)ethoxy)-1H-indole-3-formyl)phenyl)acrylamide.

[0176] ESI-MS: m / z = 378 [M+H] +;1H NMR (400MHz, DMSO-d6) δ10.35(s,1H),7.87(s,1H),7.85(s,1H),7.83(s,1H),7.74(d,J=2.5Hz,2H),7.45(s,1H),7.43(s,1H),7 .39(s,1H),7.37(s,1H),6.86(dd,J=8.8,2.5Hz,1H),6.27(d,J=2.1Hz,1H),5.73(s,1H),4.05(s,2H),3.19(s,2H),2.22(s,6H).

[0177] Example 12 N-(2-(1-(4-chlorophenyl)-2,5-dimethyl-1H-pyrrole-3-carbonyl)-5-(pyrrolidin-1-yl)phenyl)-2-(di(prop-2-yn-1-yl)amino)acetamide

[0178]

[0179] The product was synthesized by a method similar to that described in Example 11, with the addition of step d.

[0180] ESI-MS: m / z = 364 [M+H] + ;1H NMR (400MHz, DMSO-d6) δ10.65(s,-1H),10.03(s,-1H),7.47(d,J=8.2Hz,1H),7.18(s, 1H),7.16(d,J=1.9Hz,1H),7.06(d,J=2.5Hz,1H),6.97(d,J=8.0Hz,1H),6.85(d,J=2. 4Hz,1H),6.65(dd,J=8.7,2.5Hz,1H),6.36(dd,J=17.1,10.1Hz,1H),6.19(d,J=2.2Hz ,1H),5.70-5.63(m,2H),3.94(d,J=5.8Hz,2H),3.92(s,2H),3.13(s,2H),2.17(s,6H).

[0181] Example 13 4-(Dimethylamino)-N-(3-(6-methoxy-1H-indole-3-carbonyl)phenyl)but-2-enamide was synthesized according to Scheme 3, steps a, b, d.

[0182]

[0183] Step 1. Dissolve 167mg (1mmol) 3-nitrobenzoic acid in 3ml anhydrous dichloromethane, add 3 drops of N,N-dimethylformamide, then add 357mg (3mmol) thionyl chloride, and stir at room temperature for 30 minutes. TLC detection reaction is complete, and after reduced pressure concentration, a light yellow solid 3-nitrobenzoyl chloride is obtained for standby use. Dissolve 3-nitrobenzoyl chloride in 3ml anhydrous dichloromethane, then add 133mg (1mmol) aluminum chloride, and keep stirring at 0°C under nitrogen protection for 30 minutes. After the aluminum chloride solid is dissolved, add 147mg (1mmol) 5-methoxyindole at 0°C, then return to room temperature under nitrogen protection and stir for 6 hours. TLC detection reaction is complete, use DCM to extract and wash the organic phase with water for many times, dry MgSO4, and separate the crude product by flash column chromatography, with an elution gradient of EA:PE=1:2. The products were combined, concentrated and dried to obtain 221 mg of (5-methoxyindol-3-yl)(3-nitrophenyl)methanone. ESI-MS: m / z=297[M+H] + ;

[0184] Step 2. Weigh 296 mg (1 mmol) of (5-methoxyindol-3-yl) (3-nitrophenyl) ketone and dissolve it in 4 ml of methanol, then add an appropriate amount of Raney nickel and stir overnight at room temperature. TLC detection shows that the reaction is complete, and the product (3-aminophenyl) (6-methoxy-1H-indol-2-yl) ketone is obtained by filtration, concentration and drying. ESI-MS: m / z = 267 [M+H] + ;

[0185] Step 3. Weigh 12.9 mg (0.1 mmol) of 4-(dimethylamino)but-2-enoic acid hydrochloride and add it to 2 ml of dichloromethane, then add 57 mg (0.15 mmol) of 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate and 39 mg (0.3 mmol) of N,N-diisopropylethylamine in sequence, stir at room temperature for 1 hour, then add 26 mg (0.1 mmol) of (3-aminophenyl)(6-methoxy-1H-indol-2-yl)methanone, and stir at room temperature for 4 hours. TLC detection shows that the reaction is complete, and the organic phase is extracted and washed with water several times with DCM, dried with MgSO4, and the crude product is separated by flash column chromatography with an elution gradient of EA:PE=1:5. The products are combined, concentrated and dried to obtain 10 mg of (E)-4-(dimethylamino)-N-(3-(6-methoxy-1H-indole-2-carbonyl)phenyl)but-2-enoic acid amide. ESI-MS: m / z = 378 [M+H] + ; 1H NMR(400MHz, Methanol-d4)δ8.01(t,J=1.9Hz,1H),7.84(d,J=2.5Hz,1H),7.8 1(ddd,J=8.1,2.3,1.2Hz,1H),7.73(s,1H),7.50(d,J=6.7Hz,2H),7.41(t,J= 7.8Hz,1H),7.32(d,J=8.8Hz,1H),6.88(dd,J=8.8,2.5Hz,1H),6.85-6.77(m, 1H), 6.42-6.32 (m, 1H), 3.85 (s, 4H), 3.50 (dd, J = 6.8, 1.4Hz, 2H), 2.55 (s, 7H).

[0186] Example 14 4-(Dimethylamino)-N-(3-((6-methoxy-1H-indol-3-yl)methyl)phenyl)but-2-enamide

[0187]

[0188] The synthesis was carried out in a similar manner to that described in Example 13, with the addition of step c.

[0189] ESI-MS: m / z = 364 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.00(s,1H),7.73(t,J=6.4Hz,2H),7.61(d,J=2.6Hz,1H),7.29(q,J=9.0,8.2Hz,4H),6.72(dd,J =8.8, 2.6Hz, 1H), 6.60 (dt, J = 15.4, 6.2Hz, 1H), 6.26 (d, J = 15.4Hz, 1H), 3.06 (d, J = 6.2Hz, 2H), 2.11 (s, 6H), 1.59 (s, 2H).

[0190] Example 15 Determination of the inhibitory activity of compounds on USP14 protein

[0191] Compounds were added to a 384-well plate (containing 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 1 mg / mL ovalbumin, 5 mM ATP / MgCl2, 1 mM DTT (dithiothreitol), 1 nM Ptsm-VS, and 15 nM USP14) in gradient concentrations (ranging from 0.1 μM to 50 μM) and pre-incubated for 30 min. To start the reaction, 1 μM Ub-AMC was added to the system. Fluorescence was measured by real-time monitoring of the reaction at room temperature for 45 min using an Envision plate reader. Each experiment was repeated 3 times, and the average value was calculated. IC values ​​of compounds for USP14 50 The data are shown in Table 5.

[0192] Table 5

[0193] Compound No. <![CDATA[IC 50 (μM)]]> Compound No. <![CDATA[IC 50 (μM)]]> A1 25.12 A8 22.69 A2 29.51 A9 26.77 A3 3.87 A10 >50 A4 8.54 A11 >50 A5 13.28 A12 >50 A6 5.01 A13 >50 A7 >30 A14 >50

[0194] It can be seen from the above table that the compounds of the present invention can inhibit the activity of USP14 protein.

[0195] Example 16 Determination of IC of Compounds on HCT116 Cell Line 50

[0196] HCT116 cells (Institute of Cell Collection, Chinese Academy of Sciences) were inoculated in a 96-well microplate with 5000 cells per well. The compound was dissolved in DMSO and diluted to the test gradient concentration (concentration range 0.1μM-50μM). After adding all reagents, the 96-well plate was placed in an incubator for 24 hours, and then 10μl of MTT solution was added to each well, and the incubation continued for 4h. The supernatant was removed, and 150μl of DMSO was added to each well. The plate was placed on a shaker and oscillated at a low speed for 10min to fully dissolve the crystals. The absorbance of each well was measured at OD490nm in an enzyme-linked immunosorbent assay. IC inhibition of cell proliferation of the compound on the HCT116 cell line 50 As shown in Table 6.

[0197] Table 6

[0198] Compound No. <![CDATA[IC 50 (μM)]]> Compound No. <![CDATA[IC 50 (μM)]]> A1 >50 A8 >50 A2 >50 A9 >50 A3 4.61 A10 >50 A4 9.41 A11 >50 A5 37.62 A12 >50 A6 7.83 A13 >50 A7 >50 A14 >50

[0199] It can be seen from the above table that the compounds of the present invention can inhibit the proliferation of HCT116 cell line.

[0200] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A compound represented by formula (I) or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs; in, X is independently selected from R1 is selected from hydrogen, hydroxyl, amino, halogen, unsubstituted or substituted alkyl, cycloalkyl or heterocycloalkyl, and the substituent is one or more of alkyl, halogen, hydroxyl or amino; R2 is a group containing at least one of an alkenyl group, an alkynyl group, a cyano group, an acylamino group, an ethylene oxide group, and a sulfonyl fluoride group; R3 is selected from hydrogen, hydroxy, amino, dialkylamino, halogen, C 14 Alkoxy, substituted or unsubstituted alkyl, cycloalkyl, phenyl, the substituent is one or more of alkyl, halogen, hydroxyl, alkoxy, amino, nitrile, and the value of n is an integer between 1 and 6; The substituent is selected from unsubstituted or substituted aromatic heterocycles, and the substituent is selected from one or more of alkyl, halogen, hydroxyl and amino.

2. The compound according to claim 1 or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that: R1 is selected from hydrogen, hydroxy, amino, halogen, unsubstituted or substituted with 1 to 3 substituents C 1 4 Alkyl, C 3 6 Cycloalkyl or C 3 6 Heterocycloalkyl, wherein the substituent is one or more of methyl, ethyl, fluorine, hydroxyl or amino; R2 is selected from R3 is selected from hydrogen, hydroxy, amino, dimethylamino, halogen, C 1 4 Alkoxy, substituted or unsubstituted C 1 4 Alkyl, C 3 6 Cycloalkyl, phenyl, the substituent is one or more of methyl, ethyl, fluorine, chlorine, hydroxyl, methoxy, amino, nitrile, and the value of n is an integer between 1 and 4; Selected from unsubstituted or 1-3 substituent-substituted C 4-9 Aromatic heterocycle, wherein the substituent is selected from one or more of methyl, fluorine, hydroxyl and amino.

3. The compound according to claim 1 or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that: R1 is selected from hydrogen, R2 is selected from R3 is selected from Selected from 4. The compound according to claim 1 or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, pharmaceutically acceptable salt or prodrug, characterized in that: The compound has a structure shown in formula (II) or formula (III); Among them, X is selected from R1 is selected from hydrogen, R2 is selected from R3 is selected from 5. The compound according to any one of claims 1 to 4 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that: The compound is selected from:

6. A method for preparing the compound according to any one of claims 1 to 5 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, and pharmaceutically acceptable salts, characterized in that: Includes at least one of the following synthetic routes: Synthetic route 1: Synthesis route 2: Synthetic route 3: Synthetic route 4: Wherein, X, R1, and R2 have the same definitions as in the cited claims.

7. Use of the compound according to any one of claims 1 to 5 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs in the preparation of USP14 covalent inhibitors.

8. Use of the compound according to any one of claims 1 to 5 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, pharmaceutically acceptable salts or prodrugs in the preparation of anti-tumor drugs.

9. The use according to claim 7, characterized in that: The tumor is selected from one or more of cervical squamous cell carcinoma, bile duct carcinoma, hepatocellular carcinoma, gastric adenocarcinoma, sarcomatous lung cancer, thymic carcinoma, pancreatic cancer, colorectal cancer, neuroblastoma, and multiple myeloma.

10. A drug, characterized in that The invention comprises the compound according to any one of claims 1 to 5 or its stereoisomers, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates and pharmaceutically acceptable salts.

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