Benzotriazole derivatives as inhibitors of cdks, pharmaceutical compositions thereof and uses thereof

Abstract

The application belongs to the technical field of pharmaceutical chemistry, and particularly relates to a benzotriazole derivative as a CDKs inhibitor, a pharmaceutical composition thereof and application thereof in preparation of an antitumor drug. The benzotriazole derivative has strong activity in inhibiting CDK9 and other CDK subtypes, and has strong antitumor activity, and has strong inhibitory activity on cell proliferation of human blood tumor cells K562 and colorectal cancer cells HCT116. The structure of the benzotriazole derivative is shown in general formula I.

Description

Technical Field

[0001] This invention belongs to the field of medicinal chemistry technology, specifically relating to a benzotriazole derivative as a CDKs inhibitor, its pharmaceutical composition, and its application in the preparation of antitumor drugs. Background Technology

[0002] CDK9 (Cyclin-dependent kinase 9) is a member of the transcriptional CDK subfamily and plays a role in the transcriptional regulation of RNAP II. CDK9 is located on chromosome 9q34.1, and its active site has a conserved bilobal structure, consisting of an N-terminus and a C-terminus. The N-terminal lobe of CDK9 contains 16 to 108 residues, including five β-chains and a major α-helix. The C-terminal lobe contains 109 to 330 residues, including four β-chains and seven major α-helices. The hinge region of the CDK9 ATP-binding site is located in the cleft between the two lobes of the kinase. Its enzymatic activity depends on the phosphorylation of a threonine residue (Thr186) in the activated region; therefore, this ATP-binding site generally serves as a binding pocket for CDK9-related inhibitors. In terms of biological function, CDK9 is primarily involved in controlling the synthesis and processing of eukaryotic RNA polymerase II mRNA. Approximately 80% of CDK9 forms a heterodimer with Cyclin T1, while the remaining 20% ​​forms a complex with Cyclin T2A, Cyclin T2B, or Cyclin K. CDK9 activation primarily depends on the formation of the CDK9 / Cyclin T1 heterodimer, which forms the catalytic subunit of positive transcription elongation factor b (P-TEFb), thus driving transcription initiation. Inhibiting CDK9 can suppress transcriptional elongation of some genes, effectively reducing mRNA levels in tumor cells and thereby inducing tumor cell apoptosis.

[0003] Currently, there are no drugs targeting CDK9 on the market. Some investigational drugs have reached Phase III clinical trials globally, the highest stage of development globally. Among them, Alvocidib, a selective CDK9 inhibitor developed by Sanofi, has received orphan drug designation in the EU for the treatment of acute myeloid leukemia and chronic lymphocytic leukemia. Given the feasibility of CDK9-targeted drugs and referencing the successful development experience of CDK4 / 6 inhibitors, many well-known domestic (CSPC Pharmaceutical Group, Jinfang Pharmaceutical, etc.) and international (Pfizer, Bayer, Merck, AstraZeneca, etc.) companies and research institutions have already begun to invest in this target. Summary of the Invention

[0004] The purpose of this invention is to provide a benzotriazole derivative, its pharmaceutical composition, and its application. The benzotriazole derivative of this invention exhibits activity against CDK9 and other CDK subtypes (CDK2, CDK4, CDK5, CDK6, CDK7, CDK8, CDK12, CDK13, CDK15, CDK18, etc.), and its in vitro antitumor activity has been preliminarily evaluated, laying the foundation for the development and clinical application of novel antitumor drugs.

[0005] Specifically, the present invention is achieved through the following technical solutions:

[0006] In a first aspect, the present invention provides a benzotriazole derivative or a pharmaceutically acceptable salt thereof, the structure of which is shown in general formula I:

[0007]

[0008] Wherein, ring A is an aryl group containing 6-12 carbon atoms or a heteroaryl group containing 5-12 ring atoms, wherein the heteroaryl group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S;

[0009] The B ring is an aryl group containing 6-12 carbon atoms or a heteroaryl group containing 5-12 ring atoms, wherein the heteroaryl heterocycle optionally contains 1, 2, or 3 heteroatoms selected from N, O, and S; or the B ring is a cycloalkyl group containing 3-12 carbon atoms or a heterocyclic group containing 3-12 ring atoms, wherein the heterocyclic group optionally contains 1, 2, or 3 heteroatoms selected from N, O, and S; or B is a C1-C6 alkyl group or -NR. a R b R a R b Each is independently selected from hydrogen, C1-C6 alkyl, C1-C6 aminoalkyl or heterocyclic group containing 3-12 ring atoms, wherein the heterocycle of the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S;

[0010] The C ring is an aryl group containing 6-12 carbon atoms or a heteroaryl group containing 5-12 ring atoms, wherein the heteroaryl group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S.

[0011] The D ring is an aryl group containing 6-12 carbon atoms or a heteroaryl group containing 5-12 ring atoms, wherein the heteroaryl heterocycle optionally contains 1, 2, or 3 heteroatoms selected from N, O, and S; or the B ring is a cycloalkyl group containing 3-12 carbon atoms or a heterocyclic group containing 3-12 ring atoms, wherein the heterocyclic group optionally contains 1, 2, or 3 heteroatoms selected from N, O, and S; or B is a C1-C6 alkyl group or -NR. a R bR a R b Each is independently selected from hydrogen, C1-C6 alkyl, C1-C6 aminoalkyl or heterocyclic group containing 3-12 ring atoms, wherein the heterocycle of the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S;

[0012] The C ring and the D ring are fused together;

[0013] R is C1-C 12 Alkyl, C1-C 12 Alkyl groups or cycloalkyl groups containing 3-10 carbon atoms;

[0014] R 1 R 2 R 3 and R 4 Each is independently selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, hydroxyl, carboxyl, amino, cyano, nitro, C1-C6 alkylamine or cycloalkyl containing 3-10 carbon atoms;

[0015] R 5 R 6 R 7 and R 8 Each of the following groups is independently selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, hydroxyl, carboxyl, amino, cyano, nitro, C1-C6 alkylamine, C1-C6 acyl, C1-C6 sulfonyl, cycloalkyl containing 3-10 carbon atoms, or heterocyclic group containing 3-10 ring atoms, wherein the heterocycle of the heterocyclic group optionally contains 1, 2, or 3 heteroatoms selected from N, O, and S;

[0016] L represents the bond, -CR 9 R 10 -, -O-, -S-, -SO2-, -C(O)-, -NR 11 -、-SO2NR 11 -or-NR 11 SO2-;

[0017] R 9 R 10 and R 11 Each is independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, -(C1-C3 alkylene)(C3-C6 cycloalkyl), C6-C 14 Aryl, 5- to 6-membered heteroaryl, or 3- to 6-membered heterocyclic group, each of which may be independently and optionally substituted by: halogen, oxo group, -CN, -OR 12 -NR 12 R 13Or optionally substituted C1-C6 alkyl groups with halogen, -OH or oxo groups;

[0018] R 12 and R 13 Each is independently selected from hydrogen, C1-C6 alkyl groups optionally substituted with a halogen or oxo group, C2-C6 alkenyl groups optionally substituted with a halogen or oxo group, or C2-C6 alkynyl groups optionally substituted with a halogen or oxo group; or R 12 and R 13 Together with the atoms to which they are attached, they form 3- to 6-membered heterocyclic groups, which are optionally substituted with halogens, oxo groups, or C1-C6 alkyl groups that are optionally substituted with oxo groups or halogens.

[0019] Preferably, ring A is a heteroaryl group containing 5-8 ring atoms, wherein the heteroaryl group optionally contains 1 or 2 heteroatoms selected from N, O, and S.

[0020] More preferably, ring A is a heteroaryl group containing 5-6 ring atoms, wherein the heteroaryl group optionally contains one heteroatom selected from N, O, and S.

[0021] Preferably, the B ring is a cycloalkyl group containing 3-8 carbon atoms or a heterocyclic group containing 3-8 ring atoms, wherein the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S.

[0022] More preferably, the B ring is a heterocyclic group containing 5-7 ring atoms, wherein the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S.

[0023] Preferably, the C ring is a heteroaryl group containing 5-8 ring atoms, wherein the heteroaryl group optionally contains 1 or 2 heteroatoms selected from N, O, and S.

[0024] More preferably, the C ring is a heteroaryl group containing 5-6 ring atoms, wherein the heteroaryl group optionally contains one heteroatom selected from N, O, and S.

[0025] Preferably, the D ring is a cycloalkyl group containing 3-8 carbon atoms or a heterocyclic group containing 3-8 ring atoms, wherein the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S.

[0026] More preferably, the D ring is a heterocyclic group containing 5-7 ring atoms, wherein the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S.

[0027] Preferably, R is a C1-C8 alkyl or C1-C8 alkoxy.

[0028] More preferably, R is methyl, ethyl, n-propyl or isopropyl.

[0029] Preferred, R 1 R 2 R 3 and R 4 Each is independently selected from hydrogen, halogen, C1-C6 alkyl or C1-C6 alkoxy.

[0030] More preferably, R 1 R 2 R 3 and R 4 Each is independently selected from hydrogen.

[0031] Preferred, R 5 R 6 R 7 and R 8 Each is independently selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, hydroxyl, amino, C1-C6 alkylamine, C1-C6 acyl or cycloalkyl containing 3-8 carbon atoms.

[0032] More preferably, R 5 R 6 R 7 and R 8 Each is independently selected from hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 acyl or cycloalkyl containing 3-6 carbon atoms.

[0033] Preferably, L is a bond, -CR 9 R 10 -, -O-, -S-, -SO2-, -C(O-)- or -NR 11 -

[0034] More preferably, L is -C(O)-.

[0035] Preferred, R 9 R 10 and R 11 Each is independently selected from hydrogen or C1-C6 alkyl groups.

[0036] More preferably, R 9 R 10 and R 11 Each is independently selected from hydrogen.

[0037] Preferred, R 12 and R 13 Each is independently selected from hydrogen or optionally substituted with a halogen or oxo group of C1-C6 alkyl groups.

[0038] More preferably, R 12 and R 13 Each is independently selected from hydrogen.

[0039] Furthermore, the present invention preferably uses benzotriazole derivatives with the following structural formulas:

[0040]

[0041]

[0042] Furthermore, the present invention also provides a method for preparing the benzotriazole derivative represented by general formula I, the synthetic route of which is shown below:

[0043]

[0044] Where R is as defined in general formula I above; X is C; X 1 For C or N; X 2 It is hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, or halo-C1-C6 alkyl; R 14 for B and R 6 As defined in general formula I above.

[0045] In the above synthetic route, step I is a substitution reaction, in which the starting material 2-fluoro-4-bromonitrobenzene reacts with an aliphatic or cyclic amine with 1-8 carbon atoms under alkaline conditions. The reaction solvent is selected from one or more of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, dichloromethane, chloroform, toluene, tetrahydrofuran, N,N-dimethylacetamide, and 2-methyltetrahydrofuran; the base is selected from one or more of potassium carbonate, sodium carbonate, cesium carbonate, potassium hydroxide, sodium hydroxide, triethylamine, and N,N-diisopropylethylamine; and the reaction temperature is 10-100℃.

[0046] Step II is a nitro reduction reaction. The reducing agent is one or more of the following: iron powder, zinc powder, stannous chloride, sodium sulfide, sodium thiosulfate, and sodium hydrosulfite. The reaction solvent is one or more of the following: dilute hydrochloric acid (1-10 mol / L), dilute sulfuric acid (1-10 mol / L), ammonium chloride aqueous solution (1-10 mol / L), ethanol, methanol, and isopropanol. The reaction temperature is 10-100℃.

[0047] Step III is the aryltriazole ring-closing reaction, with sodium nitrite, nitrite or tert-butyl nitrite as the reactants; dilute hydrochloric acid (1-10 mol / L) and dilute sulfuric acid (1-10 mol / L) are used as the reaction solvents; the reaction temperature is -10 to 30℃.

[0048] Step IV is the Miyaura reaction, with pinacol diborate as the reactant; the catalyst is selected from [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, bis(triphenylphosphine)palladium dichloride, tetra(triphenylphosphine)palladium, or palladium acetate; the base can be one or more of potassium carbonate, sodium carbonate, cesium carbonate, potassium acetate, sodium acetate, or potassium phosphate; the reaction solvent is selected from one or more of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, toluene, tetrahydrofuran, N,N-dimethylacetamide, or 2-methyltetrahydrofuran; the protective gas is nitrogen, helium, or argon; and the reaction temperature is 10-100℃.

[0049] Step V is a Suzuki coupling reaction. The catalyst is one or more of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, bis(triphenylphosphine)palladium dichloride, tetra(triphenylphosphine)palladium, and palladium acetate. The base is one or more of potassium carbonate, sodium carbonate, cesium carbonate, potassium acetate, sodium acetate, and potassium phosphate. The reaction solvent is one or more of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, toluene, tetrahydrofuran, N,N-dimethylacetamide, and 2-methyltetrahydrofuran. The protective gas is nitrogen, helium, or argon. The reaction temperature is 10-100℃.

[0050] Step VI is the Buchwald-Hartwig reaction. The catalyst is one or more of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, bis(triphenylphosphine)palladium dichloride, tetra(triphenylphosphine)palladium, and palladium acetate. The ligand is one or more of P(o-tolyl)3, P(t-Bu)3, CyPF-t-Bu, JosiPhos, Binap, XantPhos, DPPF, BrettPhos, RuPhos, XPhos, SPhos, and BippyPhos. The base is one or more of potassium carbonate, sodium carbonate, cesium carbonate, potassium acetate, sodium acetate, and potassium phosphate. The reaction solvent is one or more of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, toluene, tetrahydrofuran, N,N-dimethylacetamide, and 2-methyltetrahydrofuran. The protective gas is nitrogen, helium, or argon. The reaction temperature is 10-120℃.

[0051] Step VII is an ester hydrolysis reaction. The base is selected from one or more of potassium carbonate, sodium carbonate, cesium carbonate, potassium hydroxide, sodium hydroxide, triethylamine, N,N-diisopropylethylamine, lithium iodide, and lithium bromide. The reaction temperature is 10-100℃. The reaction solvent is selected from one or more of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, toluene, tetrahydrofuran, N,N-dimethylacetamide, 2-methyltetrahydrofuran, methanol, ethanol, and isopropanol.

[0052] Step VIII is a condensation reaction. The condensation conditions can be DCC / EDCI / HOBt / HOAt / DMAP, HATU / HBTU / HCTU / TBTU / DIPEA / Et3N / DBU, PyBop, or T3P. The reaction solvent can be one or more of dimethyl sulfoxide, N,N-dimethylformamide, dichloromethane, acetonitrile, toluene, tetrahydrofuran, N,N-dimethylacetamide, and 2-methyltetrahydrofuran. The reaction temperature is 10-100℃.

[0053] Step IX is the Buchwald-Hartwig reaction. The catalyst is one or more of [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride, bis(triphenylphosphine)palladium dichloride, tetra(triphenylphosphine)palladium, and palladium acetate. The ligand is one or more of P(o-tolyl)3, P(t-Bu)3, CyPF-t-Bu, JosiPhos, Binap, XantPhos, DPPF, BrettPhos, RuPhos, XPhos, SPhos, and BippyPhos. The base is one or more of potassium carbonate, sodium carbonate, cesium carbonate, potassium acetate, sodium acetate, and potassium phosphate. The reaction solvent is one or more of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, toluene, tetrahydrofuran, N,N-dimethylacetamide, and 2-methyltetrahydrofuran. The protective gas is nitrogen, helium, or argon. The reaction temperature is 10-120℃.

[0054] Step X is a reductive amination reaction. The acid used in the reaction is one or more of glacial acetic acid, formic acid, dilute hydrochloric acid (1-10 mol / L), and dilute sulfuric acid (1-10 mol / L). The reducing agent is one or more of sodium borohydride, sodium cyanoborohydride, sodium triacetoxyborohydride, and sodium triisopropoxyborohydride. The reaction solvent is one or more of dimethyl sulfoxide, N,N-dimethylformamide, acetonitrile, toluene, tetrahydrofuran, N,N-dimethylacetamide, 2-methyltetrahydrofuran, methanol, ethanol, and isopropanol. The reaction temperature is 10-100℃.

[0055] In a second aspect, the present invention provides a pharmaceutical composition comprising a benzotriazole derivative of general formula I or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

[0056] In a third aspect, the present invention provides the use of a benzotriazole derivative of general formula I or a pharmaceutically acceptable salt thereof or the pharmaceutical composition thereof in the preparation of an antitumor drug.

[0057] In this invention, the term "aryl" refers to an optionally substituted monocyclic or fused bicyclic or polycyclic cyclic system having well-known aromatic characteristics, wherein at least one ring contains a fully conjugated π-electron system. Typically, an aryl group contains 6-20 carbon atoms as ring members, preferably 6-14 carbon atoms, or more preferably 6-12 carbon atoms. Examples of aryl groups include, but are not limited to, phenyl, biphenyl, naphthyl, anthraceneyl, phenanthryl, indenyl, indenyl, and tetrahydronaphthyl.

[0058] In this invention, the term "heteroaryl" refers to a monocyclic or fused bicyclic or polycyclic ring system having well-known aromatic characteristics, containing a specified number of ring atoms and including at least one heteroatom selected from N, O, and S as a ring member in the aromatic ring. The inclusion of heteroatoms allows for the aromaticity of both 5-membered and 6-membered rings. Typically, heteroaryl contains 5-20 ring atoms, preferably 5-14 ring atoms, more preferably 5-12 ring atoms. The heteroaryl ring is linked to the base molecule through the ring atoms of the heteroaryl ring, thereby maintaining aromaticity. Examples of heteroaryl often include, but are not limited to, pyrroleyl, furanyl, thiopheneyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, triazolyl, pyridinyl, pyrimidinyl, pyrazinyl, or pyridazinyl.

[0059] In this invention, the term "cycloalkyl" refers to a non-aromatic saturated carbocyclic ring system containing a specified number of carbon atoms, which can be a monocyclic, spirocyclic, bridged, or fused bicyclic or polycyclic ring system in which carbon atoms of the cycloalkyl ring are connected to the base molecule. Typically, the cycloalkyl groups of this invention contain 3-12 carbon atoms, preferably 3-8 carbon atoms. Examples of cycloalkyl groups often include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclohexane, cycloheptane, etc.

[0060] In this invention, the term "heterocyclic group" can be used interchangeably to refer to a non-aromatic saturated ring system containing a specified number of ring atoms, including at least one heteroatom selected from N, O, and S as a ring member. Typically, the heterocyclic groups of this invention contain 3-12 ring atoms, preferably 3-8 ring atoms, and more preferably 3-6 ring atoms. Examples of heterocyclic groups frequently include, but are not limited to, azirropropyl, oxacyclopropyl, thiocyclopropyl, azirrobutyl, oxacyclobutyl, thiocyclobutyl, pyrrolidinyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, tetrahydropyranyl, tetrahydrothiophenyl, morpholinyl, piperazine, etc.

[0061] In this invention, the term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0062] In this invention, the term "alkyl" (including when used alone and when included in other groups) means a branched and straight-chain saturated hydrocarbon group comprising 1-20 carbon atoms, preferably 1-12 carbon atoms, more preferably 1-5 carbon atoms, and most preferably 1-3 carbon atoms, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, 4,4-dimethylpentyl, 2,2,4-trimethylpentyl, undecyl, dodecyl, and various isomers thereof.

[0063] In this invention, the term "alkoxy" refers to an alkyl group having the stated number of carbon atoms connected by an oxygen bridge. Thus, "alkoxy" encompasses the definition of an alkyl group as described above.

[0064] In this invention, the term "alkenyl" refers to a straight-chain or branched hydrocarbon group containing a specified number of carbon atoms and at least one carbon-carbon double bond. Preferably, one carbon-carbon double bond is present, and more preferably, other parts of the compound are connected through the carbon-carbon double bond. The number of carbon atoms can be 2-12, preferably 2-5, more preferably 2, such as vinyl, 1-propenyl, 1-butenyl, etc.

[0065] In this invention, the term "alkynyl" refers to a straight-chain or branched hydrocarbon group containing a specified number of carbon atoms and at least one carbon-carbon triple bond. Preferably, one carbon-carbon triple bond is present; more preferably, the other parts of the compound are connected through the carbon-carbon triple bond. The number of carbon atoms can be 2-12, preferably 2-5, more preferably 2, such as ethynyl, 1-propynyl, 1-butynyl, etc.

[0066] In this invention, the term "aminoalkyl" refers to an alkyl group having a specified number of carbon atoms, which is substituted with one or more substituted or unsubstituted amino groups. Aminoalkyl groups typically contain 1-6 carbon atoms in the alkyl moiety and are substituted with 1, 2, or 3 amino substituents. Thus, examples of C1-C6 aminoalkyl groups often include, but are not limited to, aminomethyl (-CH2NH2), N,N-dimethylaminoethyl (-CH2CH2N(CH3)2), and 3-(N-cyclopropylamino)propyl (-CH2CH2CH2NH- c Pr) and N-pyrrolidinylethyl (-CH2CH2-N-pyrrolidinyl).

[0067] In this invention, the term "acyl" refers to a monovalent group -C(O)alkyl, wherein the alkyl moiety has a specified number of carbon atoms (typically C1-C8, preferably C1-C6 or C1-C4) and may optionally be substituted with a group suitable for the alkyl group, such as F, OH, or alkoxy. Thus, optionally substituted -C(O)C1-C4 alkyl groups include unsubstituted acyl groups, such as -C(O)CH3 (i.e., acetyl) and -C(O)CH2CH3 (i.e., propionyl), and substituted acyl groups, such as -C(O)CF3 (trifluoroacetyl), -C(O)CH2OH (hydroxyacetyl), -C(O)CH2OCH3 (methoxyacetyl), -C(O)CF2H (difluoroacetyl), etc.

[0068] Compared with the prior art, the present invention has the following advantages:

[0069] The benzotriazole derivative of this invention has a novel structure and strong inhibitory activity against CDK9 and other CDK subtypes, as well as strong antitumor activity. It also exhibits strong inhibitory activity against the proliferation of human hematologic malignancy cell line K562 and colorectal cancer cell line HCT116, laying the foundation for the development and clinical application of novel antitumor drugs. Detailed Implementation

[0070] The embodiments of the present invention are described in detail below. The embodiments are provided to better illustrate the content of the present invention and are only used to explain the present invention, and should not be construed as limiting the present invention.

[0071] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0072] Unless otherwise specified, the experimental methods used in the following embodiments are conventional methods. Unless otherwise specified, the experimental materials used in the following embodiments are commercially available products.

[0073] Example 1: (6-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-3-yl)(morpholino)methyl ketone (A-1)

[0074]

[0075] The preparation method of compound A-1, and its operation steps are as follows:

[0076] (1) 2-fluoro-4-bromonitrobenzene (10.0 g, 45.67 mmol) was dissolved in N,N-dimethylformamide, and potassium carbonate (7.63 g, 54.81 mmol) and isopropylamine (3.24 g, 54.81 mmol) were added respectively. The mixture was reacted at room temperature for 5 hours. After the reaction was completed, the reaction mixture was poured into an ice-water mixture and stirred for 2 hours. The mixture was filtered to obtain intermediate 1 as a yellow solid with a yield of 95.4%.

[0077] (2) Intermediate 1 (5 g, 19.38 mmol) was dissolved in ethanol, and reduced iron powder (5.42 g, 96.9 mmol) was added, followed by an aqueous solution of ammonium chloride (5.18 g, 96.9 mmol). The mixture was reacted at 80 °C for 1 hour. The mixture was then filtered, and the solvent was removed by rotary evaporation. The residue was redissolved in ethyl acetate and separated by silica gel column chromatography (cyclohexane / ethyl acetate = 3 / 1) to obtain intermediate 2 as a brown solid, with a yield of 89.1%.

[0078] (3) Intermediate 2 (5 g, 20.92 mmol) was dissolved in concentrated hydrochloric acid, and an aqueous solution of sodium nitrite (2.17 g, 31.38 mmol) was added. The reaction was carried out at room temperature for 3 hours. Subsequently, the pH of the system was adjusted to 8 with 1 N sodium hydroxide aqueous solution, and the mixture was extracted with ethyl acetate. The organic phases were combined and dried over anhydrous sodium sulfate. Intermediate 3 was separated by silica gel column chromatography (cyclohexane / ethyl acetate = 20 / 1) as a white solid, with a yield of 86.3%.

[0079] (4) Intermediate 3 (5 g, 20.92 mmol) was dissolved in dioxane, and pinacol diboronate (7.97 g, 31.38 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (1.53 g, 2.09 mmol), and potassium acetate (4.11 g, 41.84 mmol) were added. The mixture was reacted at 110 °C for 6 hours. After the reaction was completed, the mixture was filtered, and the filtrate was extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate and separated by silica gel column chromatography (cyclohexane / ethyl acetate = 5 / 1) to obtain intermediate 4 as a white solid, with a yield of 91.7%.

[0080] (5) Intermediate 4 (3 g, 10.45 mmol) was dissolved in a 4 / 1 mixture of dioxane and water. 2-Amino-4-bromopyridine (1.8 g, 10.45 mmol), tetrakis(triphenylphosphine)palladium (1.21 g, 1.05 mmol), and sodium carbonate (2.21 g, 20.9 mmol) were added, and the mixture was reacted at 100 °C for 3 hours. After the reaction was complete, the mixture was filtered, and the filtrate was extracted with dichloromethane. The combined organic phases were dried over anhydrous sodium sulfate and separated by silica gel column chromatography (cyclohexane / ethyl acetate = 1 / 1) to obtain intermediate 5 as a white solid, with a yield of 88.7%.

[0081] (6) Intermediate 5 (2 g, 7.90 mmol) was dissolved in N,N-dimethylformamide, and methyl 6-bromonicotinate (2.05 g, 9.48 mmol), tris(dibenzylacetone)dipalladium (0.36 g, 0.39 mmol), cesium carbonate (6.1 g, 15.8 mmol), and 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthracene (0.41 g, 0.79 mmol) were added. The reaction was carried out at 100 °C for 6 hours. After the reaction was completed, the mixture was extracted with dichloromethane, and the combined organic phases were dried over anhydrous sodium sulfate. Intermediate 6 was separated by silica gel column chromatography (cyclohexane / ethyl acetate = 1 / 1) as a yellow solid, with a yield of 65.9%.

[0082] (7) Intermediate 6 (1 g, 2.58 mmol) was dissolved in methanol, and an aqueous solution of sodium hydroxide (1 g, 25.8 mmol) was added. The reaction was carried out at 80 °C for 3 hours. After the reaction was completed, the solvent was removed by rotary evaporation, and the pH of the system was adjusted to 3 with 1 N dilute hydrochloric acid. The intermediate 7 was obtained by filtration as a white solid with a yield of 85.9%.

[0083] (8) Intermediate 7 (0.5 g, 1.34 mmol) was dissolved in N,N-dimethylformamide, and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.31 g, 1.61 mmol), 1-hydroxybenzotriazole (0.22 g, 1.61 mmol), and morpholine (0.14 g, 1.61 mmol) were added, respectively. The reaction was carried out at room temperature for 3 hours. After the reaction was completed, the mixture was extracted with dichloromethane, and the combined organic phases were dried over anhydrous sodium sulfate. The solution was separated by silica gel column chromatography (cyclohexane / ethyl acetate = 1 / 3) to obtain A-1 as a yellow solid, with a yield of 86.7%. ESI-LC-MS: 444.2 [M+H] + , 1 H NMR (600MHz, DMSO-d6) δ10.11(s,1H),8.36-8.39(m,2H),8.28(m,1H),8.18-8.20(m,1H),8.12(m,1H),7.89-7. 90(m,1H),7.73-7.79(m,2H),7.40(m,1H),5.36(hept,J=6.4Hz,1H),3.55-3.63(m,8H),1.69(d,J=6.4Hz,6H).

[0084] Example 2: (6-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-3-yl)(4-methylpiperazin-1-yl)methyl ketone (A-2)

[0085]

[0086] Compound A-2 was prepared using the same method as compound A-1, except that N-methylpiperazine was used instead of morpholine in step (8) of Example 1. A-2 was isolated as a yellow solid with a yield of 79.4%. ESI-LC-MS: 457.3 [M+H] + , 1 HNMR(600MHz,DMSO-d6)δ10.02(s,1H),8.46-8.48(m,2H),8.34(m,1H),8.17-8.19(m,1H),8.15(m,1H),7.93-8.01(m, 1H),7.83-7.89(m,2H),7.54(m,1H),5.37(hept,J=6.6Hz,1H),3.57-3.61(m,8H),2.18(s,3H),1.69(d,J=6.6Hz,6H).

[0087] Example 3: (6-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-3-yl)(4-ethylpiperazin-1-yl)methyl ketone (A-3)

[0088]

[0089] Compound A-3 was prepared using the same method as compound A-1, except that N-ethylpiperazine was used instead of morpholine in step (8) of Example 1. A-3 was isolated as a yellow solid, with a yield of 81.2%. ESI-LC-MS: 471.3 [M+H] + , 1 HNMR(600MHz,DMSO-d6)δ10.12(s,1H),8.56-8.61(m,2H),8.43(m,1H),8.27-8.29(m,1H),8.25(m,1H),8.12-8.17(m,1H),7.98-8.02 (m,2H),7.64(m,1H),5.39(hept,J=6.6Hz,1H),3.58-3.61(m,8H),2.16(q,J=4.5Hz,2H),1.70(d,J=6.6Hz,6H),1.03(t,J=4.5Hz,3H).

[0090] Example 4: (6-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-3-yl)(4-isopropylpiperazin-1-yl)methyl ketone (A-4)

[0091]

[0092] Compound A-4 was prepared using the same method as compound A-1, except that N-isopropylpiperazine was used instead of morpholine in step (8) of Example 1. A-4 was isolated as a yellow solid with a yield of 83.5%. ESI-LC-MS: 485.3 [M+H] + , 1 HNMR(600MHz,DMSO-d6)δ10.04(s,1H),8.21-8.23(m,2H),8.18(m,1H),8.16-8.17(m,1H),8.14(m,1H),8.05-8.08(m,1H),7.98-8.02 (m,2H),7.63(m,1H),5.38(hept,J=6.7Hz,1H),3.58-3.61(m,8H),2.69(m,J=5.5Hz,1H),1.70(d,J=6.5Hz,6H),1.03(d,J=5.6Hz,6H).

[0093] Example 5: (6-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-3-yl)(4-methyl-1,4-diaza-1-yl)methyl ketone (A-5)

[0094]

[0095] Compound A-5 was prepared using the same method as compound A-1, except that N-methylperiprazine was used instead of morpholine in step (8) of Example 1. A-5 was isolated as a yellow solid with a yield of 89.1%. ESI-LC-MS: 471.2 [M+H] + , 1 HNMR(600MHz,DMSO-d6)δ10.04(s,1H),8.21-8.23(m,2H),8.18(m,1H),8.16-8.17(m,1H),8.14(m,1H),8.05-8.08(m, 1H),7.98-8.02(m,2H),7.63(m,1H),5.38(hept,J=6.7Hz,1H),3.58-3.61(m,10H),2.16(s,3H),1.69(d,J=6.6Hz,6H).

[0096] Example 6: (2-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-4-yl)(morpholino)methyl ketone (A-6)

[0097]

[0098] Compound A-6 was prepared using the same method as compound A-1, except that methyl 2-bromopyridine-4-carboxylate was used instead of methyl 6-bromonicotinic acid in step (6) of Example 1. A-6 was isolated as a yellow solid with a yield of 82.3%. ESI-LC-MS: 444.2 [M+H] + , 1 H NMR (600MHz, DMSO-d6) δ10.11(s,1H),8.36-8.39(m,2H),8.28(m,1H),8.18-8.20(m,1H),8.12(m,1H),7.89-7. 90(m,1H),7.73-7.79(m,2H),7.40(m,1H),5.36(hept,J=6.4Hz,1H),3.55-3.63(m,8H),1.69(d,J=6.4Hz,6H).

[0099] Example 7: (2-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-4-yl)(4-methylpiperazin-1-yl)methyl ketone (A-7)

[0100]

[0101] Compound A-7 was prepared using the same method as compound A-1, except that methyl 2-bromopyridine-4-carboxylate was used instead of methyl 6-bromonicotinic acid in step (6) of Example 1, and N-methylpiperazine was used instead of morpholine in step (8) of Example 1. A-7 was isolated as a yellow solid with a yield of 81.9%. ESI-LC-MS: 457.3 [M+H] + , 1 H NMR(600MHz,DMSO-d6)δ10.02(s,1H),8.46-8.48(m,2H),8.34(m,1H),8.17-8.19(m,1H),8.15(m,1H),7.93-8.01(m, 1H),7.83-7.89(m,2H),7.54(m,1H),5.37(hept,J=6.6Hz,1H),3.57-3.61(m,8H),2.18(s,3H),1.69(d,J=6.6Hz,6H). Example 8: (2-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-4-yl)(4-ethylpiperazin-1-yl)methyl ketone (A-8)

[0102]

[0103] Compound A-8 was prepared in the same manner as compound A-1, except that methyl 2-bromopyridine-4-carboxylate was used instead of methyl 6-bromonicotinic acid in step (6) of Example 1, and N-ethylpiperazine was used instead of morpholine in step (8) of Example 1. A-8 was isolated as a yellow solid with a yield of 73.3%. ESI-LC-MS: 471.3 [M+H] + , 1 H NMR(600MHz,DMSO-d6)δ10.12(s,1H),8.56-8.61(m,2H),8.43(m,1H),8.27-8.29(m,1H),8.25(m,1H),8.12-8.17(m,1H),7.98-8.02( m,2H),7.64(m,1H),5.39(hept,J=6.6Hz,1H),3.58-3.61(m,8H),2.16(q,J=4.5Hz,2H),1.70(d,J=6.6Hz,6H),1.03(t,J=4.5Hz,3H).

[0104] Example 9: (2-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-4-yl)(4-isopropylpiperazin-1-yl)methyl ketone (A-9)

[0105]

[0106] Compound A-9 was prepared in the same manner as compound A-1, except that methyl 2-bromopyridine-4-carboxylate was used instead of methyl 6-bromonicotinic acid in step (6) of Example 1, and N-isopropylpiperazine was used instead of morpholine in step (8) of Example 1. A-9 was isolated as a yellow solid with a yield of 81.2%. ESI-LC-MS: 485.3 [M+H] + , 1 HNMR(600MHz,DMSO-d6)δ10.04(s,1H),8.21-8.23(m,2H),8.18(m,1H),8.16-8.17(m,1H),8.14(m,1H),8.05-8.08(m,1H),7.98-8.02 (m,2H),7.63(m,1H),5.38(hept,J=6.7Hz,1H),3.58-3.61(m,8H),2.69(m,J=5.5Hz,1H),1.70(d,J=6.5Hz,6H),1.03(d,J=5.6Hz,6H).

[0107] Example 10: (2-((4-(1-isopropyl-1H-benzo[d][1,2,3]triazol-6-yl)pyridin-2-yl)amino)pyridin-4-yl)(4-methyl-1,4-diaza-1-yl)methyl ketone (A-10)

[0108]

[0109] Compound A-10 was prepared using the same method as compound A-1, except that methyl 2-bromopyridine-4-carboxylate was used instead of methyl 6-bromonicotinic acid in step (6) of Example 1, and N-methylperipterazine was used instead of morpholine in step (8) of Example 1. A-10 was isolated as a yellow solid with a yield of 86.7%. ESI-LC-MS: 471.2 [M+H] + , 1 HNMR(600MHz,DMSO-d6)δ10.04(s,1H),8.21-8.23(m,2H),8.18(m,1H),8.16-8.17(m,1H),8.14(m,1H),8.05-8.08(m, 1H),7.98-8.02(m,2H),7.63(m,1H),5.38(hept,J=6.7Hz,1H),3.58-3.61(m,10H),2.16(s,3H),1.69(d,J=6.6Hz,6H).

[0110] Example 11: Pharmacological Activity Experiment

[0111] (1) The inhibitory effect of some compounds of the present invention on the CDK9 / Cyclin T1 complex was determined by ADP-Glo ​​method (IC50). 50 )

[0112] The DMSO stock solution and CDK9 / Cyclin T1 of the test samples were diluted to the target concentration with the appropriate buffer. The compounds of this invention were diluted in a 2-fold concentration gradient of 10 concentration points, with a maximum concentration of 500 nM. The compounds of this invention or the DMSO blank were mixed with the enzyme in a 384-well plate, and ATP and substrate were added to initiate the kinase reaction. The plate was incubated at room temperature for 60 minutes. The kinase reaction was terminated by adding ADP-Glo ​​reagent and the remaining ATP was consumed. Then, the newly generated ATP was detected by adding Kinase detection reagent. The chemiluminescence detection module of a multi-functional microplate reader was used to detect the content of newly generated ATP to reflect the enzyme activity. The results are shown in Table 1.

[0113] (2) Inhibition of cell proliferation of human hematologic malignancy cells K562 and colorectal cancer cells HCT116 using the CCK-8 assay (GI) 50 ) to conduct testing

[0114] Cells were cultured to the logarithmic growth phase using appropriate culture medium and seeded at a density of 5000 cells / well in 96-well plates. They were then incubated at 37°C for 24 hours. Afterward, DMSO stock dilutions of the test samples at appropriate concentrations were added, and blank and positive control groups were retained, with three replicates for each group. After 48 hours of incubation, 10 μL CCK-8 was added to each well and incubated for 3 hours. The absorbance at 450 nm was then measured using an Emax Microplate Reader (Molecular Devices, Sunnyvale, CA, USA). Cell viability inhibition rate was calculated based on the blank group results, as shown in Table 1.

[0115] (3) Experimental Results

[0116] Table 1. Inhibitory effects of compounds on CDK9 / Cyclin T1 complex, K562 and HCT116 cells.

[0117]

[0118] The test results in Table 1 show that the benzotriazole derivative represented by general formula I of this invention has strong inhibitory activity against CDK9 and other CDK subtypes. It is a series of CDK inhibitors with novel structures and has strong anti-tumor activity, exhibiting strong inhibitory activity against the proliferation of human hematologic malignancy cells K562 and colorectal cancer cells HCT116. It has potential application prospects and clinical research value.

[0119] Obviously, the above embodiments are merely examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those skilled in the art can make various modifications and variations to the present invention without departing from the spirit and scope of the invention. Therefore, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention also intends to include these modifications and variations.

Claims

1. A benzotriazole derivative or a pharmaceutically acceptable salt thereof, characterized in that, The structure of the benzotriazole derivative is shown in general formula I: General Formula I in, Ring A is a pyridinyl group; The B ring is a cycloalkyl group containing 3-12 carbon atoms or a heterocyclic group containing 3-12 ring atoms, wherein the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O or S; R is C1-C 12 Alkyl, C1-C 12 Alkyl groups or cycloalkyl groups containing 3-10 carbon atoms; R 1 R 2 R 3 and R 4 Each is independently selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, hydroxyl, carboxyl, amino, cyano, nitro, C1-C6 alkylamine or cycloalkyl containing 3-10 carbon atoms; R 6 The group is independently selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, hydroxyl, carboxyl, amino, cyano, nitro, C1-C6 alkylamine, C1-C6 acyl, C1-C6 sulfonyl, cycloalkyl containing 3-10 carbon atoms, or heterocyclic group containing 3-10 ring atoms, wherein the heterocycle of the heterocyclic group optionally contains 1, 2, or 3 heteroatoms selected from N, O, and S; L is -C(O)-.

2. The benzotriazole derivative of general formula I according to claim 1, characterized in that, In general formula I, ring B is a cycloalkyl group containing 3-8 carbon atoms or a heterocyclic group containing 3-8 ring atoms, wherein the heterocyclic group optionally contains 1, 2 or 3 heteroatoms selected from N, O, and S; R is a C1-C8 alkyl group or a C1-C8 alkoxy group; R 1 R 2 R 3 and R 4 Each is independently selected from hydrogen, halogen, C1-C6 alkyl, or C1-C6 alkoxy; R 6 It is independently selected from hydrogen, halogen, C1-C6 alkyl, C1-C6 alkoxy, halogenated C1-C6 alkyl, hydroxyl, amino, C1-C6 alkylamine, C1-C6 acyl or cycloalkyl containing 3-8 carbon atoms.

3. The benzotriazole derivative of general formula I according to claim 2, characterized in that, In general formula I, ring B is a heterocyclic group containing 5-7 ring atoms, wherein the heterocyclic group optionally contains 1, 2, or 3 heteroatoms selected from N, O, and S; R is methyl, ethyl, n-propyl, or isopropyl; R 1 R 2 R 3 and R 4 Each is independently selected from hydrogen; R 6 It is independently selected from hydrogen, halogen, C1-C6 alkyl, halogenated C1-C6 alkyl, C1-C6 acyl or cycloalkyl containing 3-6 carbon atoms.

4. A benzotriazole derivative or a pharmaceutically acceptable salt thereof, characterized in that, The structural formula of the benzotriazole derivative is shown below:

5. A pharmaceutical composition, characterized in that, It comprises the benzotriazole derivative as described in any one of claims 1-4, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

6. The use of the benzotriazole derivative of any one of claims 1-4 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition of claim 5 in the preparation of an antitumor drug.

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