Ring A is selected from 3-8 membered cycloalkyl or 3-8 membered heterocyclyl optionally substituted with 1-2 substituents selected from halogen, hydroxy, amino, nitro, cyano, C_1-6Alkyl radical, C_1-6Alkylamino radical, di (C)_1-6Alkyl) amino, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Alkylthio radical, C_1-6Alkylcarbonyl, halo C_1-6Alkoxy, halo C_1-6Alkylthio, hydroxy C_1-6Alkoxy, hydroxy C_1-6Alkylthio, amino C_1-6Alkoxy, amino C_1-6An alkylthio group;

R²、R³、R⁴each independently selected from hydrogen, halogen, nitro, cyano, amino, hydroxyl and C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, cyano C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, hydroxy C_1-6Alkoxy, amino C_1-6Alkoxy or cyano C_1-6An alkoxy group;

R⁵、R⁶each independently selected from hydrogen and C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl or cyano C_1-6An alkyl group.

In certain embodiments, ring a is selected from a 3-6 membered cycloalkyl or 3-6 membered heterocyclyl optionally substituted with 1-2 substituents; the substituent is selected from halogen, hydroxyl, amino, nitro, cyano and C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl radical, C_1-6Alkoxy or halo C_1-6An alkoxy group.

In certain embodiments, ring a is selected from 5-6 membered saturated cycloalkyl or 5-6 membered saturated heterocyclyl optionally substituted with 1-2 substituents; the substituent is selected from halogen, hydroxyl, amino, nitro, cyano and C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl radical, C_1-6Alkoxy or halo C_1-6An alkoxy group.

In certain embodiments, ring a is selected from cyclohexyl, tetrahydrofuranyl, oxacyclohexanyl or azacyclohexanyl, optionally substituted with 1-2 substituents; the substituents are selected from fluoro, chloro, hydroxy, methyl, ethyl, propyl, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, trifluoromethoxy or trifluoroethoxy.

In certain embodiments, ring a is selected from cyclohexyl, 1-difluorocyclohexyl, 1-hydroxy-cyclohexyl, 1-hydroxy-1-methyl-cyclohexyl, 1-dimethylcyclohexyl, oxacyclohexyl, or azacyclohexyl.

In certain embodiments, ring A is selected from cyclohexyl, 1-difluorocyclohex-4-yl, 1-hydroxy-cyclohex-4-yl, 1-hydroxy-1-methyl-cyclohex-4-yl, 1-dimethylcyclohex-4-yl, oxacyclohex-4-yl, or azacyclohex-4-yl.

In certain embodiments, R¹Selected from hydrogen or C_1-6An alkyl group;

X₁is selected from C (R)²) Or N;

X₂is selected from N (R)⁵)；

X is selected from-S (O) -, -S (O)₂-or-S (O) (NR)⁶)-；

Ring A is selected from 3-8 membered rings optionally substituted with 1-2 substituentsAlkyl or 3-8 membered heterocyclyl, said substituents being selected from halogen, hydroxy, amino, nitro, cyano, C_1-6Alkyl radical, C_1-6Alkylamino radical, di (C)_1-6Alkyl) amino, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Alkylthio radical, C_1-6Alkylcarbonyl, halo C_1-6Alkoxy, halo C_1-6Alkylthio, hydroxy C_1-6Alkoxy, hydroxy C_1-6Alkylthio, amino C_1-6Alkoxy, amino C_1-6An alkylthio group;

R²selected from hydrogen, halogen, nitro, cyano, amino, hydroxy, C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl, cyano C_1-6Alkyl radical, C_1-6Alkoxy, halo C_1-6Alkoxy, hydroxy C_1-6Alkoxy, amino C_1-6Alkoxy or cyano C_1-6An alkoxy group;

In certain embodiments, R¹Selected from hydrogen or C_1-6An alkyl group;

X₁is selected from C (R)²) Or N;

X₂is selected from N (R)⁵)；

X is selected from-S (O) -, -S (O)₂-or-S (O) (NR)⁶)-；

Ring A is selected from 5-6 membered saturated cycloalkyl or 5-6 membered saturated heterocyclyl optionally substituted with 1-2 substituents selected from halogen, hydroxy, amino, nitro, cyano, C_1-6Alkyl radical, C_1-6Alkylamino radical, di (C)_1-6Alkyl) amino, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Alkylthio radical, C_1-6Alkylcarbonyl, halo C_1-6Alkoxy radicalHalogen substituted C_1-6Alkylthio, hydroxy C_1-6Alkoxy, hydroxy C_1-6Alkylthio, amino C_1-6Alkoxy, amino C_1-6An alkylthio group;

In certain embodiments, R¹Selected from hydrogen or C_1-6An alkyl group;

X₁is selected from C (R)²) Or N;

X₂is selected from N (R)⁵)；

X is selected from-S (O) -, -S (O)₂-or-S (O) (NR)⁶)-；

Ring A is selected from 5-6 membered saturated cycloalkyl or 5-6 membered saturated heterocyclyl;

In certain embodiments, the compound of formula (I), a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, further has a structure represented by formula (II),

wherein, X, X₁、R²、R⁵、R⁶And ring a is as previously described.

In certain embodiments, X₁Is selected from C (R)²) Or N;

R²selected from hydrogen, halogen, cyano, amino, hydroxy, C_1-6Alkyl, halo C_1-6Alkyl radical, C_1-6Alkoxy or halo C_1-6An alkoxy group.

In certain embodiments, X₁Is selected from C (R)²) Or N;

R²selected from hydrogen, fluorine, chlorine, hydroxyl, methyl, methoxy, trifluoromethyl and trifluoromethoxy.

In certain embodiments, R⁵Selected from hydrogen, C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl or amino C_1-6An alkyl group.

In certain embodiments, R⁵Selected from hydrogen, methyl, monofluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl or aminomethyl.

In certain embodiments, R⁵Selected from hydrogen, methyl, monofluoromethyl, difluoromethyl or trifluoromethyl.

In certain embodiments, X is selected from the group consisting of-C (O) -, -S (O)₂-or-S (O) (NR)⁶)-；R⁶Selected from hydrogen, methyl, monofluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl or aminomethyl.

In certain embodiments X is selected from the group consisting of-C (O) -, -S (O)₂-or-S (O) (NR)⁶)-；R⁶Selected from hydrogen, methyl, trifluoromethyl, hydroxymethyl or aminomethyl.

In certain embodiments, X is selected from the group consisting of-C (O) -, -S (O)₂-or-S (O) (NR)⁶)-；R⁶Selected from hydrogen or methyl.

In certain embodiments, ring A is selected from the group consisting of oxacyclohex-4-yl.

wherein, X₁Is selected from C (R)²) Or N;

x is selected from the group consisting of-C (O) -, -S (O)₂-or-S (O) (NR)⁶) And when X is-C (O) -X is₁Is C (R)²)；

In certain embodiments, X₁Is selected from C (R)²) Or N;

R²selected from hydrogen, fluoro, chloro, hydroxy, methyl, methoxy, trifluoromethyl, trifluoromethoxy;

R⁵selected from hydrogen, methyl, monofluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl or aminomethyl;

R⁶Selected from hydrogen, methyl, monofluoromethyl, difluoromethyl, trifluoromethyl, hydroxymethyl or aminomethyl;

ring A is selected from 5-6 membered saturated cycloalkyl or 5-6 membered saturated heterocyclyl optionally substituted with 1-2 substituents; the substituent is selected from halogen, hydroxyl, amino, nitro, cyano and C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl radical, C_1-6Alkoxy or halo C_1-6An alkoxy group.

In certain embodiments, X₁Is selected from C (R)²) Or N;

ring A is selected from the group consisting of cyclohexyl, 1-difluorocyclohexyl, 1-hydroxy-cyclohexyl, 1-hydroxy-1-methyl-cyclohexyl, 1-dimethylcyclohexyl, oxacyclohexyl and azacyclohexyl.

In certain embodiments, X₁Is N;

x is selected from-S (O) -, -S (O)₂-or-S (O) (NR)⁶)-；

In certain embodiments, X₁Is N;

x is selected from-S (O) -or-S (O)₂-；

R⁵selected from hydrogen, C_1-6Alkyl, halo C_1-6Alkyl, hydroxy C_1-6Alkyl, amino C_1-6Alkyl or cyano C_1-6An alkyl group.

In certain embodiments, X₁Is N;

x is selected from-S (O) -or-S (O)₂-；

Ring A is selected from a 5-6 membered saturated cycloalkyl group or a 5-6 membered saturated heterocyclyl group.

In certain embodiments, X₁Is N;

x is selected from-S (O) -or-S (O)₂-；

The technical solutions of the present invention can be combined with each other to form a new technical solution, and the formed new technical solution is also included in the scope of the present invention.

In certain embodiments, the compound of formula (I), a pharmaceutically acceptable salt thereof, or an isomer thereof, is selected from the compounds shown in table 1 below:

TABLE 1

In another aspect, the present invention also provides a pharmaceutical preparation, which contains the compound described in the above general formulas (I), (II), its pharmaceutically acceptable salt or its isomer, and one or more pharmaceutically acceptable excipients, and the pharmaceutical preparation can be any pharmaceutically acceptable dosage form. Pharmaceutically acceptable excipients are substances which are non-toxic, compatible with the active ingredient and otherwise biologically suitable for use in the organism. The choice of a particular excipient will depend on the mode of administration or disease type and state used to treat a particular patient.

In certain embodiments, the pharmaceutical formulations described above may be administered to a patient or subject in need of such treatment by oral, parenteral, rectal, or pulmonary administration, among others. For oral administration, the pharmaceutical composition can be prepared into oral preparations, for example, conventional oral solid preparations such as tablets, capsules, pills, granules and the like; it can also be made into oral liquid, such as oral solution, oral suspension, syrup, etc. When the composition is formulated into oral preparations, appropriate filler, binder, disintegrating agent, lubricant, etc. can be added. For parenteral administration, the above pharmaceutical preparations may also be prepared into injections, including injections, sterile powders for injection and concentrated solutions for injection. The injection can be prepared by conventional method in the existing pharmaceutical field, and can be prepared without adding additives or adding suitable additives according to the properties of the medicine. For rectal administration, the pharmaceutical composition may be formulated as a suppository or the like. For pulmonary administration, the pharmaceutical composition may be formulated as an inhalation formulation, aerosol, powder spray, or the like.

In another aspect, the present invention also relates to the use of the compound of the aforementioned general formula (I), (II), its pharmaceutically acceptable salt or its isomer in the preparation of a medicament for preventing and/or treating diseases such as benign tumor or cancer, including carcinoma in situ and metastatic carcinoma.

Furthermore, the invention also relates to application of a pharmaceutical preparation containing the compounds of the general formulas (I) and (II), the pharmaceutically acceptable salts thereof or the isomers thereof in preparing a medicament for preventing and/or treating diseases such as benign tumors or cancers, wherein the cancers comprise carcinoma in situ and metastatic cancers.

In another aspect, the present invention also relates to the use of the compound of the aforementioned general formula (I), (II), its pharmaceutically acceptable salt or its isomer in the preparation of a medicament for preventing and/or treating diseases such as benign tumor or cancer, in combination with radiotherapy and/or one or more anticancer agents, wherein the cancer includes carcinoma in situ and metastatic cancer.

Furthermore, the invention also relates to the application of a pharmaceutical preparation containing the compound shown in the general formula (I) and the general formula (II), the pharmaceutically acceptable salt thereof or the isomer thereof in preparing a medicament for preventing and/or treating diseases such as benign tumors or cancers, wherein the medicament can be combined with radiotherapy and/or one or more anticancer agents, and the cancers comprise carcinoma in situ and metastatic cancers.

In another aspect, the present invention also relates to the use of the compound of the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof, or an isomer thereof, for the preparation of a medicament for sensitizing cancer cells to anticancer agents and/or ionizing radiation.

Furthermore, the invention also relates to application of a pharmaceutical preparation containing the compound shown in the general formula (I) and the general formula (II), the pharmaceutically acceptable salt thereof or the isomer thereof in preparing a medicament for sensitizing cancer cells to anticancer agents and/or ionizing radiation.

The ionizing radiation refers to the radiation of various energy rays received by a patient during the radiotherapy process.

In another aspect, the present invention also provides a pharmaceutical composition comprising a compound of the foregoing general formula (I), (II), a pharmaceutically acceptable salt thereof, or an isomer thereof, and one or more second therapeutically active agents selected from the group consisting of anti-cancer agents, including mitotic inhibitors, alkylating agents, anti-metabolites, DNA chimerics, anti-tumor antibiotics, growth factor inhibitors, signaling inhibitors, cell cycle inhibitors, enzyme inhibitors, retinoid receptor modulators, proteasome inhibitors, topoisomerase inhibitors, biological response modifiers, hormonal agents, angiogenesis inhibitors, cell growth inhibitors, targeting antibodies, HMG-CoA reductase inhibitors, and prenyl protein transferase inhibitors.

In certain embodiments, the second therapeutically active agent can be a drug that reduces or reduces one or more side effects of a compound of the invention when used to treat a disease in a subject, or can enhance the efficacy of a compound of the invention.

In certain embodiments, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients, as described above.

In another aspect, the present invention also relates to the use of a pharmaceutical composition containing the compound of the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof, or an isomer thereof, in the preparation of a medicament for preventing and/or treating diseases such as benign tumor or cancer, including carcinoma in situ and metastatic carcinoma.

In another aspect, the present invention also relates to the use of a pharmaceutical composition containing the compound of the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof or an isomer thereof in the preparation of a medicament for preventing and/or treating diseases such as benign tumor or cancer, which can be used in combination with radiotherapy and/or one or more anticancer agents, wherein the cancer includes carcinoma in situ and metastatic cancer.

Furthermore, the invention also relates to application of a pharmaceutical composition containing the compound shown in the general formula (I) and the general formula (II), the pharmaceutically acceptable salt thereof or the isomer thereof in preparing a medicament for sensitizing cancer cells to anticancer agents and/or ionizing radiation.

In another aspect, the present invention also provides a method for treating a disease associated with DNAPK overactivation, the method comprising administering to a patient in need thereof an effective amount of a compound of the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof, or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition; the disease associated with DNAPK over-activation is selected from benign tumors or cancers, including carcinoma in situ and metastatic carcinoma.

Further, the present invention provides a method for treating a disease associated with DNAPK overactivation, which comprises administering an effective amount of a compound represented by the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition to a patient before/after receiving radiotherapy; the disease associated with DNAPK over-activation is selected from benign tumors or cancers, including carcinoma in situ and metastatic carcinoma.

Further, the present invention provides a method for treating a disease associated with DNAPK overactivation, comprising administering an effective amount of a compound represented by the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition to a patient before/after receiving chemotherapy; the disease associated with DNAPK over-activation is selected from benign tumors or cancers, including carcinoma in situ and metastatic carcinoma.

In another aspect, the present invention also provides a method for enhancing the sensitivity of a patient to an anticancer agent or radiation therapy, which comprises administering to a patient in need thereof an effective amount of a compound represented by the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition; the anticancer agent is as described below.

Further, the present invention provides a method for enhancing the sensitivity of a patient to an anticancer agent or radiation therapy, which comprises administering an effective amount of a compound represented by the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition to the patient before/after receiving radiation therapy; the anticancer agent is as described below.

Further, the present invention provides a method for enhancing the sensitivity of a patient to an anticancer agent or radiation therapy, which comprises administering an effective amount of a compound represented by the aforementioned general formula (I), (II), a pharmaceutically acceptable salt thereof or an isomer thereof, the aforementioned pharmaceutical preparation or pharmaceutical composition to the patient before/after receiving chemotherapy; the anticancer agent is as described below.

In another aspect, the present invention also provides a kit comprising:

(a) an effective amount of one or more compounds of the general formulas (I) and (II), pharmaceutically acceptable salts or isomers thereof,

and (b) an effective amount of one or more anti-cancer agents.

The term "anticancer agent" as used herein refers to an agent having a certain preventive, palliative, retarding, inhibitory or curative effect on tumors, including but not limited to mitotic inhibitors, alkylating agents, antimetabolites, DNA chimerics, antitumor antibiotics, growth factor inhibitors, signal transduction inhibitors, cell cycle inhibitors, enzyme inhibitors, retinoid receptor modulators, proteasome inhibitors, topoisomerase inhibitors, biological response modifiers, hormonal drugs, angiogenesis inhibitors, cell growth inhibitors, targeting antibodies, HMG-CoA reductase inhibitors, prenyl protein transferase inhibitors, etc.; the tumor includes benign tumor and cancer. By "effective amount" is meant a dosage of a drug that prevents, alleviates, retards, inhibits or cures a condition in a subject. The size of the administered dose is related to the administration mode of the drug, the pharmacokinetics of the medicament, the severity of the disease, the individual signs (sex, weight, height, age) of the subject, and the like.

In the present invention, unless otherwise defined, scientific and technical terms used herein have meanings commonly understood by those skilled in the art, however, in order to better understand the present invention, definitions of some terms are provided below. To the extent that the definitions and explanations of terms provided herein do not conform to the meanings commonly understood by those skilled in the art, the definitions and explanations of terms provided herein shall control.

The "halogen" as referred to herein means a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

"C" according to the invention_1-6Alkyl "denotes straight or branched alkyl having 1 to 6 carbon atoms, including for example" C_1-4Alkyl group "," C_1-3Alkyl group "," C_1-2Alkyl group "," C_2-6Alkyl group "," C_2-5Alkyl group "," C_2-4Alkyl group "," C_2-3Alkyl group "," C_3-6Alkyl group "," C_3-5Alkyl group "," C_3-4Alkyl "and the like, specific examples include, but are not limited to: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, 2-methylbutyl, neopentyl, 1-ethylpropyl, n-hexyl, isohexyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 1, 2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 1, 2-dimethylpropyl, and the like. "C" according to the invention_1-4Alkyl "means C_1-6Specific examples of the alkyl group having 1 to 4 carbon atoms.

"C" according to the invention_1-6Alkoxy "means" C_1-6alkyl-O- ", said" C_1-6Alkyl "is as defined above. "C" according to the invention_1-4Alkoxy "means" C_1-4alkyl-O- ", said" C_1-4Alkyl "is as defined above.

"C" according to the invention_1-6Alkylthio "means" C_1-6alkyl-S- ", said" C_1-6Alkyl "is as defined above. "C" according to the invention_1-4Alkylthio "means" C_1-4alkyl-S- ", said" C_1-4Alkyl "is as defined above.

The "hydroxy group C" of the present invention_1-6Alkyl, amino C_1-6Alkyl, halo C_1-6Alkyl, cyano C_1-6Alkyl "means C_1-6One or more hydrogens of the alkyl group are each replaced by one or more hydroxy, amino, halogen or cyano groups. C_1-6Alkyl is as previously defined

The "hydroxy group C" of the present invention_1-6Alkoxy, amino C_1-6Alkoxy, halo C_1-6Alkoxy, cyano C_1-6Alkoxy "means" C_1-6One or more hydrogens of "alkoxy" are replaced with one or more hydroxy, amino, halo, or cyano groups.

Described in the invention "Hydroxy radical C_1-6Alkylthio, amino C_1-6Alkylthio, halo C_1-6Alkylthio "means" C_1-6Alkylthio "is one in which one or more hydrogens are replaced with one or more hydroxy, amino, or halogen.

"C" according to the invention_1-6Alkylamino radical, di (C)_1-6Alkyl) amino "means independently C_1-6alkyl-NH-),

The "3-to 8-membered cycloalkyl" as referred to herein means a saturated or partially saturated monocyclic cyclic group having 3 to 8 ring atoms and having no aromaticity, and the "3-to 8-membered cycloalkyl" as referred to herein includes "3-to 8-membered saturated cycloalkyl" and "3-to 8-membered partially saturated cycloalkyl", and is exemplified by "3-to 6-membered cycloalkyl", "3-to 6-membered saturated cycloalkyl", "5-to 6-membered saturated cycloalkyl", and the like. Examples include, but are not limited to: cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl or cyclohexenyl and the like.

The "3-to 8-membered heterocyclic group" as used herein means a saturated or partially saturated and non-aromatic monocyclic cyclic group containing at least one (e.g., 1,2, 3,4 or 5) hetero atom which is a nitrogen atom, an oxygen atom and/or a sulfur atom and has 3 to 8 ring atoms, and optionally, a ring atom (e.g., a carbon atom, a nitrogen atom or a sulfur atom) in the cyclic structure may be oxo. The "3-to 8-membered heterocyclic group" described in the present invention includes "3-to 8-membered saturated heterocyclic group" and "3-to 8-membered partially saturated heterocyclic group". The "3-to 8-membered heterocyclic group" is exemplified by "3-to 6-membered heterocyclic group", "3-to 6-membered saturated heterocyclic group", "3-to 5-membered saturated heterocyclic group", "5-to 6-membered saturated heterocyclic group", etc. Specific examples thereof include, but are not limited to: aziridinyl, 2H-aziridinyl, diazacyclopropenyl, 3H-diazacyclopropenyl, azetidinyl, oxetanyl, 1, 4-dioxanyl, 1, 3-dioxanyl, 1, 4-dioxadienyl, tetrahydrofuranyl, dihydropyrrolyl, tetrahydropyrrolyl, tetrahydropyrazolidinyl, tetrahydroimidazolyl, 4, 5-dihydroimidazolyl, pyrazolidinyl, 4, 5-dihydropyrazolyl, 2, 5-dihydrothienyl, tetrahydrothienyl, 4, 5-dihydrothiazolyl, thiazolidinyl, tetrahydropyranyl (oxetanyl), tetrahydrothiopyranyl, 1-dioxotetrahydrothiopyranyl, piperidinyl (azacyclohexanyl), Tetrahydropyridinyl, piperidonyl, tetrahydropyridonyl, dihydropiperidonyl, piperazinyl, hexahydropyrimidyl, morpholinyl and the like.

"C (R) according to the invention²)”、“C(R³)(R⁴)”、“N(R⁵) "refers to the following structures, respectively:

the present invention is said to be "-C (O) -", "-S (O)")₂-”、“-S(O)(NR⁶) - "denotes the following structure, respectively:

represents the position of attachment of the group to an adjacent group.

The term "optionally substituted with a substituent" as used herein refers to both the case where one or more hydrogen atoms on a substituted group are "substituted" or "unsubstituted" with one or more substituents.

The chemotherapy is the abbreviation of chemical drug therapy, and achieves the purpose of treatment mainly by using chemical therapeutic drugs to kill cancer cells.

The "radiotherapy" in the invention refers to a tumor treatment method, i.e. tumor radiotherapy, which mainly uses radioactive rays to perform local tumor treatment, wherein the "radioactive rays" include alpha, beta and gamma rays generated by radioactive isotopes, and x rays, electron beams, proton beams and other particle beams generated by various x-ray treatment machines or accelerators.

"pharmaceutically acceptable salt" as used herein refers to an acidic functional group (e.g., -COOH, -OH, -SO) present in a compound₃H, etc.) with a suitable inorganic or organic cation (base), including salts with alkali or alkaline earth metals, ammonium salts, salts with nitrogen-containing organic bases; and salts of basic functional groups present in the compounds (e.g., -NH2, etc.) with suitable inorganic or organic anions (acids), including salts with inorganic or organic acids (e.g., carboxylic acids, etc.).

"isomers" as used herein means that the compounds of the present invention contain one or more asymmetric centers and thus are available as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The compounds of the present invention may have asymmetric centers that each independently produce two optical isomers. The scope of the present invention includes all possible optical isomers and mixtures thereof. The compounds of the present invention, if they contain an olefinic double bond, include cis-isomers and trans-isomers, unless otherwise specified. The compounds of the invention may exist in tautomeric (one of the functional group isomers) forms having different points of attachment of hydrogen through one or more double bond shifts, e.g., a ketone and its enol form are keto-enol tautomers. The compounds of the present invention contain a spiro ring structure, and substituents on the ring may be present on both sides of the ring to form the opposite cis (cis) and trans (trans) isomers, depending on the steric structure of the ring. Each tautomer and mixtures thereof are included within the scope of the present invention. All enantiomers, diastereomers, racemates, meso, cis-trans isomers, tautomers, geometric isomers, epimers, mixtures thereof and the like of the compounds are included within the scope of the present invention.

The compounds of the invention may be prepared by enantiospecific synthesis or by resolution from a mixture of enantiomers in such a way as to give the individual enantiomers. Conventional resolution techniques include resolving mixtures of enantiomers of the starting material or the final product using various well-known chromatographic methods.

When the stereochemistry of the disclosed compounds is named or depicted by structure, the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% pure by weight relative to the other stereoisomers. When a single isomer is named or depicted by structure, the depicted or named enantiomer is at least 60%, 70%, 80%, 90%, 99%, or 99.9% pure by weight. The optical purity wt% is the ratio of the weight of an enantiomer to the weight of the enantiomer plus the weight of its optical isomer.

Advantageous effects of the invention

1. The compound, the pharmaceutically acceptable salt or the isomer thereof has excellent DNA-PK inhibitory effect, has good pharmacokinetic property in organisms, has lasting effect and high bioavailability, and can enhance the sensitivity of cancer cells to radiotherapy and/or one or more anticancer agents.

2. The compound, the pharmaceutically acceptable salt or the isomer thereof has good inhibition effect on benign tumors and cancers, and the stability of the liver microsome is high.

3. The compound of the invention has simple preparation process, high medicine purity, stable quality and easy large-scale industrial production.

Detailed description of the preferred embodiments

The technical solutions of the present invention will be described below in conjunction with the specific embodiments, and the above-mentioned contents of the present invention will be further described in detail, but it should not be understood that the scope of the above-mentioned subject matter of the present invention is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Abbreviations:

BrettPhos Pd G3: methanesulfonic acid (2-dicyclohexylphosphino-3, 6-dimethoxy-2 ',4',6' -triisopropyl-1, 1' -biphenyl) (2-amino-1, 1' -biphenyl-2-yl) palladium (II); pd₂(dba)₃: tris (dibenzylideneacetone) dipalladium; xantphos: 4, 5-bis (diphenylphosphino) -9, 9-bisMethyl xanthene; TMEDA: tetramethylethylenediamine; PE: petroleum ether; EA is ethyl acetate; DIEA: n, N-diisopropylethylamine.

The first embodiment is as follows: preparation of 7-fluoro-3-methyl-6- ((7-methyl- [1,2,4] triazolo [1,5-a ] pyridin-6-yl) amino) -1- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro-2H-imidazo [4,5-c ] pyridin-2-one (compound 1)

1. Preparation of tert-butyl (6-chloro-5-fluoropyridin-3-yl) carbamate

5-bromo-2-chloro-3-fluoropyridine (25.0g,118.8mmol), BocNH₂(15.3g，130.7mmol)，Pd₂(dba)₃(5.4 g, 5.9mmol), Xantphos (4.8g, 8.3mmol) and Cs₂CO₃(77.4g, 237.6mmol) of 1, 4-dioxane (500.0mL), N₂Stirring at 85 deg.C for 6.0 hr under protection. The product was obtained by silica gel column chromatography (PE: EA ═ 10:1 to 3:1) (18.1g, yield: 61.8%).

2. Preparation of tert-butyl (6-chloro-5-fluoro-4-iodopyridin-3-yl) carbamate

Under the protection of nitrogen, 2.5Mn-BuLi (38.9mL,97.2mmol) was slowly added dropwise to a solution of tert-butyl (6-chloro-5-fluoropyridin-3-yl) carbamate (8.0g,32.4mmol) and TMEDA (11.3g,97.2mmol) in tetrahydrofuran (100mL) at-78 deg.C, the mixture was stirred for 90 minutes while heating to-20 deg.C, then cooled to-78 deg.C, a solution of iodine (24.7g, 97.2mmol) in tetrahydrofuran (50.0mL) was slowly added, and the mixture was stirred for 6 hours while slowly heating to 30 deg.C. Water (150.0mL) was added thereto, and the mixture was quenched, extracted with ethyl acetate (200.0mL), and subjected to silica gel column chromatography (PE: EA ═ 5:1 to 1:1) to give a product (12.0g, yield: 99.2%).

3. Preparation of 6-chloro-7-fluoro-1- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro-2H-imidazo [4,5-c ] pyridin-2-one

Tert-butyl (6-chloro-5-fluoro-4-iodopyridin-3-yl) carbamate (11.0g,29.5mmol), 4-aminotetrahydropyran hydrochloride (4.1g,29.5mmol), Pd₂(dba)₃(2.2g,2.4mmol), Xantphos (2.0g,3.5mmol) and Cs₂CO₃(28.8g,88.5mmol) of 1, 4-dioxane (500.0mL), N₂Stirring at 100 deg.C for 12.0 hr under protection. The product was obtained by silica gel column chromatography (PE: EA ═ 1:1) (3.0g, yield: 37.5%).

4. Preparation of 6-chloro-7-fluoro-3-methyl-1- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro-2H-imidazo [4,5-c ] pyridin-2-one

To a solution of 6-chloro-7-fluoro-1- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro-2H-imidazo [4,5-c ] pyridin-2-one (400.0mg,1.5mmol) in DMF (10.0mL) at 0 ℃ was added 60% NaH (240.0mg, 6.0mmol), reacted at 26 ℃ for 0.5 hour, then iodomethane (1.1g,7.5mmol) was added, the system was concentrated after reacted at 26 ℃ for 1.0 hour, then water (5.0mL) was added, and ethyl acetate (10.0mL) was added and extracted to give the product (410.0mg, yield: 97.1%).

5. Preparation of 7-fluoro-3-methyl-6- ((7-methyl- [1,2,4] triazolo [1,5-a ] pyridin-6-yl) amino) -1- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro-2H-imidazo [4,5-c ] pyridin-2-one

Mixing 6-chloro-7-fluoro-3-methyl-1- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro-2H-imidazo [4, 5-c)]Pyridin-2-one (400.0mg,1.4mmol), 7-methyl- [1,2,4]Triazolo [1,5-a]Pyridin-6-amine (207.5mg,1.4mmol), cesium carbonate (912.2mg,2.8mmol) and Brettphos Pd G3(127.2mg,0.14mmol) were added to 1, 4-dioxane (10.0mL) in a N-flask₂Reacting at 100 deg.C for 6.0 hr, concentrating, and passing through reversed phase column layerThe product was obtained by separation (acetonitrile, water) (168.6mg, yield: 30.3%).

The molecular formula is as follows: c₁₉H₂₀FN₇O₂Molecular weight: 397.4 LC-MS (M/e): 398.2(M + H +)

¹HNMR(400MHz,DMSO):δ8.99(s,1H),8.35(s,1H),8.05(s,1H),7.75(s,1H),7.70 (s,1H),4.55-4.60(m,1H),3.98-4.02(m,2H),3.47-3.50(m,2H),3.33(s,3H),2.33(s,3H), 1.99-2.22(m,2H),1.74-1.77(m,2H)

Example two: preparation of 1-methyl-5- ((7-methyl- [1,2,4] triazolo [1,5-a ] pyridin-6-yl) amino) -3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazol [3,4-d ] pyrimidine 2, 2-dioxide (Compound 4)

1. Preparation of 2-chloro-5-nitro-N- (tetrahydro-2H-pyran-4-yl) pyrimidin-4-amine

2, 4-dichloro-5-nitropyrimidine (10g,51.5mmol) was dissolved in dichloromethane (200mL) and DIEA (16.6g,129mmol), tetrahydro-2H-pyran-4-amine hydrochloride (7.1g,51.5mmol) were added sequentially at-70 ℃ and after completion the temperature was raised naturally to 25 ℃. Concentration and column chromatography (ethyl acetate/n-heptane 0-40%) gave the product (11g, 83% yield).

2. 2-chloro-N⁴Preparation of (tetrahydro-2H-pyran-4-yl) pyrimidine-4, 5-diamine

Dissolving 2-chloro-5-nitro-N- (tetrahydro-2H-pyran-4-yl) pyrimidin-4-amine (3g,11.6mmol) in ethanol (60mL), adding stannous chloride (6.6g,34.8mmol), reacting at 80 ℃ for 2H, concentrating the reaction solution, diluting with ethyl acetate, adding saturated aqueous sodium bicarbonate solution, precipitating a solid, filtering with celite, extracting the filtrate with ethyl acetate to obtain an ethyl acetate phase, drying with anhydrous sodium sulfate, filtering, concentrating, and performing column chromatography (50% -90% ethyl acetate/N-heptane) to obtain a product (1.67g, yield 63%).

3. 2-chloro-N⁵-methyl-N⁴Preparation of (tetrahydro-2H-pyran-4-yl) pyrimidine-4, 5-diamine

2-chloro-N⁴- (tetrahydro-2H-pyran-4-amine) pyrimidine-4, 5-diamine (1.67g,7.3mmol) was dissolved in methanol (30mL), paraformaldehyde (monomer molecular weight 30,438mg,14.6mmol), sodium methoxide (2.0g,36.5mmol) were added, the mixture was sealed at 50 ℃ for 4 hours, cooled to room temperature, added with sodium borohydride (691mg,18.3mmol), reacted at 25 ℃ for 1 hour, concentrated, and subjected to column chromatography (ethyl acetate/n-heptane ═ 50% to 90%) to obtain a product (1.2g, yield 68%).

4. Preparation of 5-chloro-1-methyl-3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazole [3,4-d ] pyrimidine 2-oxide

2-chloro-N⁵-methyl-N⁴- (tetrahydro-2H-pyran-4-yl) pyrimidine-4, 5-diamine (800mg,3.3mmol) was dissolved in chloroform (25 mL), thionyl chloride (2.5mL) was added, reacted at 70 ℃ for 3 hours, and concentrated to give the product (750mg, yield 79%).

5. Preparation of 5-chloro-1-methyl-3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazole [3,4-d ] pyrimidine 2, 2-dioxide

5-chloro-1-methyl-3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazole [3,4-d ] pyrimidine 2-oxide (100 mg,0.35mmol) was dissolved in dichloromethane/water (3/3mL), benzyltriethylammonium chloride (40mg,0.18mmol), 10% aqueous sodium hypochlorite solution (1.4mL) was added, reacted at 25 ℃ for 2 hours, ethyl acetate was extracted twice, the ethyl acetate phase was dried over anhydrous sodium sulfate, filtered, concentrated, and subjected to column chromatography (30% -50% ethyl acetate/n-heptane) to obtain a product (60mg, yield 57%).

6. Preparation of 1-methyl-5- ((7-methyl- [1,2,4] triazolo [1,5-a ] pyridin-6-yl) amino) -3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazole [3,4-d ] pyrimidine 2, 2-dioxide

5-chloro-1-methyl-3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazole [3,4-d ] pyrimidine 2, 2-dioxide (60mg,0.2mmol) was dissolved in dioxane (10mL), 7-methyl- [1,2,4] triazolo [1,5-a ] pyridin-6-amine (35 mg,0.24mmol), Brettphos Pd-G3(55mg,0.06mmol), cesium carbonate (156mg,0.48mmol) was added, reacted at 80 ℃ for 4 hours, concentrated, and column chromatographed (acetonitrile/water 0-45%) to give the product (39mg, 47% yield).

Molecular formula C₁₇H₂₀N₈O₃S molecular weight 416.5 LC-MS (M/e):437.2(M + H)⁺)

¹H-NMR(400MHz,CDCl₃)δ:9.68(s,1H),8.26(s,1H),7.66(s,1H),7.57(s,1H),6.62 (s,1H),4.52-4.38(m,1H),4.21-4.10(m,2H),3.60-3.50(m,2H),3.29(s,3H),2.68-2.55(m, 2H),2.49(s,3H),2.08-1.98(m,2H).

Example three: preparation of 1-methyl-5- ((7-methyl- [1,2,4] triazolo [1,5-a ] pyridin-6-yl) amino) -3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazol [3,4-d ] pyrimidine 2-oxide (Compound 5)

5-chloro-1-methyl-3- (tetrahydro-2H-pyran-4-yl) -1, 3-dihydro- [1,2,5] thiadiazole [3,4-d ] pyrimidine 2-oxide (50 mg,0.17mmol) was dissolved in dioxane (10mL), 7-methyl- [1,2,4] triazolo [1,5-a ] pyridin-6-amine (31 mg,0.21mmol), BrettPhos Pd-G3(15mg,0.017mmol), cesium carbonate (133mg,0.41mmol) was added, reacted at 80 ℃ for 4 hours, concentrated, and column chromatographed (methanol/water 1/15) to give the product (10.8mg, 16% yield).

Molecular formula C₁₇H₂₀N₈O₂Molecular weight of S400.5 LC-MS (M/e):401.2(M + H +)

¹H-NMR(400MHz,CDCl₃)δ:9.37(s,1H),8.30(s,1H),7.79(s,1H),7.61(s,1H), 4.60-4.40(m,4H),4.12-4.01(m,2H),3.60-3.45(m,2H),2.50(s,3H),2.45-2.00(m,4H).

The following compounds were prepared using the same or similar methods as in the above examples:

experimental protocol

An exemplary experimental scheme of a portion of the compounds of the invention is provided below to show the advantageous activity and advantageous technical effects of the compounds of the invention. It should be understood, however, that the following experimental protocols are only illustrative of the present disclosure and are not intended to limit the scope of the present disclosure.

Experimental example 1 in vitro enzymatic Activity of the Compound of the present invention

Abbreviations

EDTA: ethylenediaminetetraacetic acid

DMSO, DMSO: dimethyl sulfoxide

Tris (Tris): tris (hydroxymethyl) aminomethane

Brij-35: polyoxyethylene lauryl ether

DTT: dithiothreitol

And (3) testing the sample: the compounds of table 1 of the present invention, their structural formulae and methods of preparation are shown in the examples.

Experimental reagent:

name (R)	Brand
		ADP-Glo Kinase Assay	Promege
DNA-PK	Promege

The experimental method comprises the following steps:

1. 1-fold kinase buffer solution is prepared

1) 1-fold kinase buffer

40mM Tris,pH 7.5

0.0055％Brij-35

20mM MgCl₂

0.05mM DTT

2. Compound preparation

1) The initial concentration of the compound to be tested was 1. mu.M, and the compound was prepared at 100-fold concentration, i.e., 100. mu.M. Mu.l of 10mM compound was taken and 198. mu.l of 100% DMSO was added to prepare a 100. mu.M solution of the compound. 100 μ l of 100-fold compound was added to the second well of the 96-well plate, and 60 μ l of 100% DMSO was added to the other wells. Mu.l of compound from the second well was added to the third well and diluted sequentially 3-fold further down for a total of 10 concentrations.

2) Transfer the highest concentration (400nM) of 100. mu.l of 100% DMSO and the positive control Wortmannin to two empty wells as Max and Min wells, respectively.

3) Echo was used to transfer 50nl of compound to 384-well plates.

3. Preparation of 2 Xkinase solution

1) A2-fold DNA-PK kinase solution was prepared using a 1-fold kinase buffer.

2) Transfer 2.5. mu.l of 2-fold enzyme solution to 384-well reaction wells.

3) Shaking, mixing, and standing at room temperature.

4. Preparation of 2 Xsubstrate solution

1) A2-fold substrate solution was prepared using 1-fold kinase buffer.

2) Transfer 2.5. mu.l of 2-fold substrate solution to 384-well reaction wells to initiate the reaction.

3) Oscillating and mixing.

5. Kinase reaction and termination

1) The 384 well plates were capped and incubated at 28 ℃ for 3 hours.

2) Transfer 5. mu.l ADP-Glo reagent and incubate at 28 ℃ for 2 hours.

6. Detection of reaction results

1) The reaction was stopped by transferring 10. mu.l of the kinase detection reagent to reaction wells of a 384-well plate.

2) Rest for 30 minutes at room temperature.

7. Data reading

Sample values were read at Envision.

8. Inhibition rate calculation

1) Data is copied from Envision.

2) This was converted to inhibition data.

Percent inhibition is (max-conversion)/(max-min) 100. where max refers to the conversion rate of the DMSO control, min refers to the conversion rate of the no enzyme control, and conversion refers to the conversion rate at each concentration of test compound.

3) Data were imported into MS Excel and curve-fitted using XLFit Excel add-in version 5.4.0.8.

The experimental results are as follows:

the compounds of Table 1 of the present invention have good DNA-PK inhibitory activity, IC thereof₅₀The values were all between 1-300nM, as shown in Table 2 below.

TABLE 2 in vitro enzymatic Activity data for Compounds of the invention

Experimental example 2 Metabolic stability of liver microsomes in various species of the Compound of the present invention

And (3) testing the sample: the chemical name and the preparation method of the compound are shown in the preparation examples of each compound.

Experimental materials:

human mixed liver microsomes (purchased) with a liver microsomed protein concentration of 20 mg/mL^-1。

Beagle mixed liver microsomes (purchased), the concentration of liver microsomes protein is 20 mg/mL^-1。

Rat Mixed liver microsomes (purchased), liver microsomes protein concentration 20 mg. multidot.mL^-1。

Experimental initiator β -NADPH (purchased); phosphate buffered saline (PBS, manufactured by Takara Shuzo) at pH 7.4.

Preparing a test solution:

a proper amount of test powder is precisely weighed, a proper amount of dimethyl sulfoxide (DMSO) is added to dissolve the test powder to 1mM, and the test powder is diluted by 20 times to 50 mu M of working solution by using methanol.

The experimental method comprises the following steps:

TABLE 3 liver microsome metabolic stability experiment incubation system composition

The experimental operation steps are as follows:

(1) according to the above Table 3 "constitution of the Experimental incubation System", 5.85 mL of 100mM PBS, 20mM MgCl were used for each compound₂Solution 0.585mL and H₂O3.57 mL, and a mixed solution 1 (not containing microsomes, a sample and. beta. -NADPH) for incubation was prepared. The positive pair drug verapamil of the experimental incubation system was also performed to demonstrate normal liver microsomal enzyme activity.

(2) Liver microsomes (20mg protein/mL) were removed from the-80 ℃ freezer and placed on a 37 ℃ water bath constant temperature shaker for pre-incubation for 3 min.

(3) For each compound, 1.9mL of mixed solution 1 of incubation system was taken for each species, and 56. mu.L of microsomes of different species was added to prepare mixed solution 2 of incubation system (containing no test substance and. beta. -NADPH).

(4) Sample set (microsome and β -NADPH containing): and adding 14 mu L of the test sample working solution with the concentration of 50 mu M into 616 mu L of the mixed solution 2 of the incubation system, and adding 70 mu L of 10mM beta-NADPH working solution. Mixing, and repeating the steps. The sampling time points are 0min, 5min, 10min, 20min, 30min and 60 min. This sample set was used to evaluate the metabolic stability of compounds mediated via β -NADPH.

(5) Control group (microsome-containing, no β -NADPH, water instead of β -NADPH): 264 mu L of the mixed solution 2 of the incubation system is taken, 6 mu L of the working solution of the test article with the concentration of 50 mu M is added, and 30 mu L of water is added. Mixing, and repeating the steps. Sampling time points were 0min and 60 min. This negative control group was used to evaluate whether compounds present non- β -NADPH mediated metabolism in the liver microsome incubation system.

(6) At each predetermined time point, 50 μ L of sample was taken from the incubation sample tube, added to a stop sample tube (containing 300 μ L of cold stop reagent, containing 50ng/mL acetonitrile as internal standard of tolbutamide), vortexed, and the reaction was stopped.

(7) After vortexing for 10min, centrifuge for 5min (12000 rpm).

(8) Taking 100 mu L of supernatant, adding 100 mu L of water, mixing uniformly by vortex, and carrying out LC-MS/MS sample injection analysis.

And (3) data analysis:

the percent residual was converted by the ratio of the peak area of the test article to the internal standard in the following equation.

The experimental results are as follows:

TABLE 4 hepatic microsome stability results for the compounds of the invention

And (4) experimental conclusion:

the compounds of the invention have good stability in human, canine and rat liver microsomes.

Experimental example 3 CD1 mouse pharmacokinetics experiment of the Compound of the present invention

And (3) testing the sample: the chemical name and the preparation method of the compound are shown in the preparation examples of each compound. The test animals were: CD1 mouse, female, 6 mice/route of administration.

Preparing a test solution:

the preparation method of the blank solvent (1) comprises the following steps: weighing 28g of HP-beta-CD, adding a proper amount of water for injection to dissolve, then fixing the volume to 100mL by using the water for injection, and uniformly mixing by vortex to obtain 28% HP-beta-CD.

The preparation method of the blank solvent (2) comprises the following steps: weighing 20g of HPC, slowly adding 500mL of stirred purified water, then adding 1mL of Tween 80, stirring until the mixture is clear and transparent, diluting to 1000mL, and uniformly stirring to obtain 2% of HPC + 0.1% of Tween 80.

IV (bolus IV) administration:

weighing the compound 4(1.95mg) of the invention, adding DMA (54.3 mu L), carrying out vortex dissolution, then adding PEG400(90.5 mu L), carrying out vortex mixing, finally adding a blank solvent (1) (1.665mL), carrying out vortex mixing, and carrying out heat preservation at 50 ℃ for 20min to prepare a 1mg/mL clear solution as an IV administration solution of the test compound.

PO (intragastric) administration:

the compound (19.49mg) of the embodiment of the invention is weighed and placed in a tissue grinder, a blank solvent (2) (3.61mL) is added, the mixture is uniformly ground at the rotating speed of 1200 r/min, and suspension liquid medicine of 5mg/mL is prepared and is used as PO administration liquid medicine of the test compound.

Experimental methods

The IV dose is 5mg/kg, the administration concentration is 1mg/mL, and the administration volume is 5 mL/kg.

PO was administered at a dose of 50mg/kg, at a concentration of 5mg/mL, and at a volume of 10 mL/kg.

Blood sampling time points: blood was collected at 0.083, 0.25, 0.5, 1,2,4, 6, 8, 24h after administration, specifically in the manner shown in the following table:

approximately 50. mu.L of whole blood was collected at each time point via the canthus and placedTo contain EDTA-K₂Centrifuging at 8000 rpm at 4 deg.C for 6min in an anticoagulant tube to obtain plasma sample, and freezing at-80 deg.C in a refrigerator for analysis.

Plasma sample analysis

Adopting a protein precipitation method: taking 20 mu L of a plasma sample, adding 200 mu L of an internal standard (acetonitrile solution containing 50ng/mL of tolbutamide), vortexing for 10min, then centrifuging for 20min at 4000 rpm, taking 100 mu L of supernatant, then adding 100 mu L of water, vortexing and uniformly mixing for 3min, and then analyzing the drug concentration in the plasma by LC-MS/MS.

Results of the experiment

TABLE 5 evaluation results of CD1 mouse PK

AUC_0-tArea under curve 0 → t when drug is represented; CL represents clearance; v_ssRepresenting the steady state apparent distribution volume; t is_1/2Represents a terminal elimination half-life; t is_maxRepresents the time to peak; c_max(ii) surrogate expression peak concentration; f% represents the absolute bioavailability.

Conclusion of the experiment

The test result shows that the compound has good pharmacokinetic property and higher exposure and bioavailability.

Claims

1. A compound represented by the general formula (I), a pharmaceutically acceptable salt thereof or an isomer thereof,

wherein,

R¹selected from hydrogen or C_1-6An alkyl group;

X₁is selected from C (R)²) Or N;

X₂is selected from C (R)³)(R⁴) Or N (R)⁵)；

2. The compound, pharmaceutically acceptable salt thereof, or isomer thereof according to claim 1,

ring A is selected from 3-6 membered cycloalkyl or 3-6 membered heterocyclyl optionally substituted with 1-2 substituents;

preferably, ring A is selected from 5-6 membered saturated cycloalkyl or 5-6 membered saturated heterocyclyl optionally substituted with 1-2 substituents;

the substituent is selected from halogen, hydroxyl, amino, nitro, cyano and C_1-6Alkyl, halo C_1-6Alkyl radicalHydroxy group C_1-6Alkyl, amino C_1-6Alkyl radical, C_1-6Alkoxy or halo C_1-6An alkoxy group.

3. The compound, a pharmaceutically acceptable salt thereof, or an isomer thereof according to any one of claims 1 to 2,

ring A is selected from cyclohexyl, tetrahydrofuranyl, oxacyclohexanyl or azacyclohexanyl, optionally substituted with 1-2 substituents; the substituents are selected from fluoro, chloro, hydroxy, methyl, ethyl, propyl, monofluoromethyl, difluoromethyl, trifluoromethyl, trifluoroethyl, methoxy, ethoxy, trifluoromethoxy or trifluoroethoxy.

4. The compound of any one of claims 1-3, a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, further having a structure represented by the following general formula (II),

wherein, X, X₁、R²、R⁵、R⁶And ring A is as defined in any one of claims 1 to 3.

5. The compound, a pharmaceutically acceptable salt thereof, or an isomer thereof according to any one of claims 1 to 4,

X₁is selected from C (R)²) Or N;

x is selected from the group consisting of-C (O) -, -S (O)₂-or-S (O) (NR)⁶)-；

6. The compound of claim 1, a pharmaceutically acceptable salt thereof, or an isomer thereof, selected from the group consisting of:

7. a pharmaceutical formulation comprising a compound according to any one of claims 1 to 6, a pharmaceutically acceptable salt thereof or an isomer thereof, wherein the pharmaceutical formulation comprises one or more pharmaceutically acceptable excipients, and wherein the pharmaceutical formulation is in any pharmaceutically acceptable dosage form.

8. A pharmaceutical composition comprising a compound of any one of claims 1-6, a pharmaceutically acceptable salt thereof, or an isomer thereof, comprising one or more second therapeutically active agents selected from the group consisting of anti-cancer agents, including mitotic inhibitors, alkylating agents, anti-metabolites, DNA chimerics, anti-tumor antibiotics, growth factor inhibitors, signaling inhibitors, cell cycle inhibitors, enzyme inhibitors, retinoid receptor modulators, proteasome inhibitors, topoisomerase inhibitors, biological response modifiers, hormonal agents, angiogenesis inhibitors, cell growth inhibitors, targeting antibodies, HMG-CoA reductase inhibitors, and prenyl protein transferase inhibitors.

9. Use of a compound according to any one of claims 1 to 6, a pharmaceutically acceptable salt or isomer thereof, a pharmaceutical formulation according to claim 7, or a pharmaceutical composition according to claim 8 for the manufacture of a medicament for the prevention and/or treatment of benign tumours or cancers, including carcinoma in situ and metastatic cancers.

10. Use of a compound according to any one of claims 1 to 6, a pharmaceutically acceptable salt or isomer thereof, a pharmaceutical formulation according to claim 7, or a pharmaceutical composition according to claim 8 in the manufacture of a medicament for the prevention and/or treatment of benign tumours or cancers, including carcinoma in situ and metastatic carcinoma, in combination with radiotherapy and/or one or more anti-cancer agents.

11. Use of a compound of any one of claims 1-6, a pharmaceutically acceptable salt or isomer thereof, a pharmaceutical formulation of claim 7, or a pharmaceutical composition of claim 8, in the manufacture of a medicament for sensitizing cancer cells to an anti-cancer agent and/or radiation therapy.