POL theta inhibitor and application thereof

Abstract

The invention relates to a POL theta inhibitor as shown in a formula I, wherein the definition of each substituent group in # imgabs0 # is as shown in the specification. The invention also relates to a composition containing the compound or the salt thereof and application of the compound or the salt thereof in preparation of medicines for treating Pol theta-mediated diseases.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to a POLθ inhibitor and uses thereof. Background Art

[0002] DNA double strand breaks (DSBs) are one of the most serious DNA damages. In this context, DNA double strand breaks (DSBs) repair plays a key role in cell growth and cancer development. In particular, tumor cells with defects in DNA repair have a selective growth advantage, leading to genetic instability and promoting tumor evolution, but they often over-rely on alternative repair pathways, which represents a tumor cell-specific vulnerability that can be targeted for killing tumor cells (Pilié, PGet al. Nat. Rev. Clin. Oncol. 2019, 16(2), 81-104.). Mammalian cells use four repair pathways to repair highly cytotoxic DNA double strand breaks (DSBs) to maintain genome integrity and cell viability: non-homologous end joining (NHEJ), homologous recombination (HR), single strand annealing (SSA) and alternative end joining (alt-EJ) or microhomology-mediated end joining (MMEJ). There are two different genetic mechanisms of MMEJ, among which DNA polymerase θ (POLθ)-mediated end joining (TMEJ) is the main mechanism (Huang, R. et al. Signal Transduct. Target. Ther. 2021, 6(1), 254.).

[0003] The clinical success of PARPi provides proof of principle for the synthetic lethal approach. Although PARPi has encouraging therapeutic benefits, these drugs have shown some key problems, such as poor clinical efficacy, ineffectiveness against certain DNA repair-deficient cancers, and the development of acquired and innate resistance. The main resistance mechanism observed in preclinical and clinical models is related to the reversal mutation of BRCA2, which allows the correct encoding of functional proteins, thereby restoring the HR pathway. For these reasons, there is an urgent need to identify other synthetic lethal pathways involved in the DNA repair pathway to develop new treatments for BRCA-deficient tumors (Dias, MP et al. Nat. Rev. Clin. Oncol. 2021, 18 (12), 773-791.).

[0004] Among synthetic lethal targets, POLθ (encoded by the PolQ gene) is particularly promising for the treatment of BRCA1-deficient cancers. It is barely expressed in normal tissues but highly expressed in a variety of tumor types (such as breast cancer, ovarian cancer, head and neck squamous cell carcinoma, and lung cancer), and its overexpression is associated with poor prognosis. POLθ belongs to the A family polymerase and is a large (2590 residues in humans) multifunctional protein that contains a superfamily II N-terminal conserved helicase-like domain (called POLθ-hel, residues 32-899) and a C-terminal conserved DNA polymerase domain (called POLθ-pol, residues 1819-2590), connected by an unstructured central region. Both POLθ-hel and POLθ-pol domains are critical for TMEJ activity, and the central portion may have a regulatory role (Black, SJ et al. Nat. Commun. 2019, 10(1), 4423.).

[0005] In summary, POLθ is crucial for cells with homologous recombination defects, and there is currently an unmet need for the treatment of homologous recombination-deficient tumors. Inhibiting POLθ can inhibit microhomology-mediated end-joining repair in cells. The development of POLθ inhibitors could provide a novel strategy for the targeted treatment of homologous recombination-deficient tumors. Summary of the Invention

[0006] The present invention provides a class of compounds having POLθ inhibitory activity.

[0007] The present invention relates to a compound of the following formula I, a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

[0008]

[0009] wherein R1 and R2 are each independently selected from H, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl;

[0010] R3 is selected from H, amino, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0011] or said R3 and R1 or R2 and adjacent N and C form a heterocyclic group, said heterocyclic group is optionally substituted by hydrogen, halogen, amino, hydroxy, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy; R4 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, carbonyl, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclic group, aryl, heteroaryl;

[0012] R5 is each independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0013] B is aryl or heteroaryl, said aryl or heteroaryl being optionally substituted with hydrogen, halogen, amino, hydroxy, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy;

[0014] n is an integer from 0 to 4.

[0015] In some embodiments, R3 is aryl or heteroaryl, wherein the heteroaryl group contains at least one atom selected from N, O, and S.

[0016] In some embodiments, the present invention relates to a compound of Formula II, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0017]

[0018] wherein A1, A2, A3, and A4 are each independently selected from CR6, CR6R6a, NR6, O, S, and S(O)2; and R6 is independently selected from hydrogen, hydroxy, halogen, alkyl, haloalkyl, alkoxy, amino, aminoalkyl, acyl, cycloalkyl, aryl, heterocyclyl, and heteroaryl;

[0019] or A1 and A2, A2 and A3 optionally form a substituted or unsubstituted saturated or unsaturated 3-14 membered ring, which may optionally include one or more heteroatoms, which may be the same or different and are independently selected from O, N, and S;

[0020] R6a is independently selected from hydrogen, hydroxy, halogen, alkyl, haloalkyl, alkoxy, amino, aminoalkyl, acyl; or R6 and R6a together with the carbon atom to which they are attached form a -C(=S)-, (-C)=O, -C(=NH)- group, or a substituted or unsubstituted saturated or unsaturated 3-14 membered ring which may optionally include one or more heteroatoms which may be the same or different and which are independently selected from O, N, and S;

[0021] B is aryl or heteroaryl, said aryl or heteroaryl being optionally substituted with hydrogen, hydroxy, halogen, haloalkyl, alkoxy, amino, aminoalkyl, acyl, alkyl, cycloalkyl, aryl, heterocyclyl and heteroaryl;

[0022] R4 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, carbonyl, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0023] R5 is each independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl.

[0024] In some embodiments, Selected from:

[0025]

[0026] In some embodiments, the present invention relates to a compound of Formula III, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0027]

[0028] in:

[0029] A1 is selected from CR6, (-C)=O, S(O)2, R6 is independently selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl;

[0030] A2 is selected from CR6, NR6, O, R6 is independently selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl;

[0031] Or A1 and A2 form a 5-6 membered aryl or heteroaryl group;

[0032] R7 is selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl;

[0033] The other substituents are as defined above.

[0034] In some embodiments, B is selected from a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group may optionally include one or more heteroatoms, which may be the same or different and independently selected from O, N, and S; and the aryl or heteroaryl group is optionally substituted with hydrogen, hydroxyl, halogen, cyano, alkyl, haloalkyl, alkoxy, amino, or aminoalkyl.

[0035] In some embodiments, B is selected from:

[0036]

[0037] in:

[0038] A5, A6, A7, A8 are CR11 or NR11, R11 is independently selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl; R8 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl.

[0039] In some embodiments, B is selected from:

[0040]

[0041] R8 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl.

[0042] In some embodiments, the present invention relates to a compound of Formula IV, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0043]

[0044] R8 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl

[0045] R4 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, carbonyl, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl;

[0046] R5 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl.

[0047] In some embodiments, R8 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl.

[0048] In some embodiments, R4 is selected from hydrogen, hydroxyl, halogen, amino, carbonyl, C1-6 alkyl, C2-6 alkenyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-6 cycloalkyl, C3-6 heterocyclyl.

[0049] In some embodiments, R5 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl.

[0050] In some embodiments, the present invention relates to a compound of Formula V, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0051]

[0052] Formula V:

[0053] R8 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl.

[0054] R9 and R10 are each independently selected from hydrogen, hydroxyl, halogen, amino, carbonyl, C1-6 alkyl, C2-6 alkenyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-6 cycloalkyl, C3-6 heterocyclyl, and the C3-6 heterocyclyl may optionally include one or more O, N and S heteroatoms.

[0055] R5 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl.

[0056] The present invention provides a compound having the following structure, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof:

[0057]

[0058]

[0059]

[0060] In another aspect, the present invention provides a pharmaceutical composition comprising the above-mentioned compound, its stereoisomers or pharmaceutically acceptable salts thereof and one or more pharmaceutically acceptable carriers or excipients.

[0061] On the other hand, the present invention provides use of the above-mentioned compound, its stereoisomer or pharmaceutically acceptable salt, or the above-mentioned pharmaceutical composition in the preparation of a medicament for treating Polθ-mediated diseases.

[0062] In some embodiments, the Pol θ-mediated disease is liver cancer, breast cancer, ovarian cancer, lung cancer, kidney cancer, prostate cancer, skin cancer, bladder cancer, pancreatic cancer, or head and neck cancer.

[0063] Regarding the technical effects achieved by the present invention, on the one hand, the compounds of the present invention show an IC50 of less than 100nM in terms of the inhibitory effect on Pol θ. In some preferred embodiments, the compounds of the present invention show an IC50 of less than 50nM in terms of the inhibitory effect on Pol θ. In some preferred embodiments, the compounds of the present invention show an IC50 of less than 10nM in terms of the inhibitory effect on Pol θ. In some preferred embodiments, the IC50 of some compounds is 1-100nM. In some preferred embodiments, the IC50 of some compounds is 1-50nM.

[0064] Terminology

[0065] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0066] In the present invention, an alkyl group refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched-chain isomers thereof.

[0067] More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.

[0068] The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available point of attachment. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate groups. Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl and hydroxy-substituted alkyl are preferred in the present invention; the hydroxy-substituted alkyl may be 2-hydroxyisopropyl or 1-hydroxyethyl.

[0069] In the present invention, the cycloalkyl group refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, and the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 carbon atoms, and further preferably 3 to 6 carbon atoms.

[0070] Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; and polycyclic cycloalkyls include spirocycloalkyls, fused cycloalkyls, and bridged cycloalkyls.

[0071] In the present invention, spiroalkyl refers to a polycyclic group sharing a carbon atom (called spiro atom) between 5 to 20 monocycles, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. According to the number of spiro atoms shared between the rings, spiroalkyl is divided into single spiroalkyl, double spiroalkyl or multiple spiroalkyl, preferably single spiroalkyl and double spiroalkyl. More preferably, it is 4 / 4 members, 4 / 5 members, 4 / 6 members, 5 / 5 members or 5 / 6 members of single spiroalkyl.

[0072] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, etc. The cycloalkyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0073] In the present invention, heterocyclic group refers to a saturated or partially unsaturated monocyclic or polycyclic heterocyclic group containing 3 to 20 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, but excluding the ring portion of -OO-, -OS- or -SS-, the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 10 ring atoms; and further preferably, it contains 3 to 8 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, pyrrolidonyl, piperidin-2-onyl, 3,4-dihydropyridin-2(1H)-onyl, 4,5-dihydropyridazin-3(2H)-onyl, azetidinyl, oxetanyl, oxanyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc.; preferably pyrrolidyl, pyrrolidonyl, piperidin-2-one, 3,4-dihydropyridin-2 (1H) -one, 4,5-dihydropyridazine-3 (2H) -one, azetidinyl, oxetanyl, dihydropyrrolyl, tetrahydrofuranyl, pyrazolidinyl, morpholinyl, Piperazinyl and pyranyl; more preferably dihydropyrrolyl, pyrrolidinyl, pyrrolidonyl, piperidin-2-onyl, 3,4-dihydropyridin-2 (1H) -onyl, 4,5-dihydropyridazin-3 (2H) -onyl, azetidinyl, oxetanyl, oxanyl, morpholinyl, piperidinyl, piperazinyl, Pyranyl. Polycyclic heterocyclic groups include spirocyclic, fused-ring, and bridged heterocyclic groups; wherein the spirocyclic, fused-ring, and bridged heterocyclic groups are optionally connected to other groups through single bonds, or further connected to other cycloalkyl, heterocyclic, aryl, and heteroaryl groups through any two or more atoms on the ring.

[0074] The heterocyclyl ring may be fused to an aryl, heteroaryl, or cycloalkyl ring, wherein the ring attached to the parent structure is the heterocyclyl ring.

[0075] In the present invention, aryl refers to a 5- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 5- to 10-membered, more preferably 5- to 8-membered, such as phenyl and naphthyl, preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl, or cycloalkyl ring, wherein the ring connected to the parent structure is the aryl ring, non-limiting examples of which include:

[0076]

[0077] The aryl group may be substituted or unsubstituted. When substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0078] In the present invention, heteroaryl refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 yuan, more preferably 5 to 8 yuan, most preferably 5 yuan or 6 yuan, such as pyrazinyl, pyridazinyl, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, oxadiazole, pyrazinyl etc., preferably pyrimidinyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazole, pyridine. The heteroaryl ring can be fused on an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, and its non-limiting examples include:

[0079]

[0080] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0081] In the present invention, alkoxy refers to-O-(alkyl) and-O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as described above, preferably containing an alkyl of 1 to 8 carbon atoms, more preferably an alkyl of 1 to 6 carbon atoms, most preferably an alkyl of 1 to 3 carbon atoms. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents are preferably one or more following groups, independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0082] In the present invention, haloalkyl refers to an alkyl group substituted by one or more halogens, wherein alkyl is as defined above. Non-limiting examples of haloalkyl include: trifluoromethyl, trifluoroethyl;

[0083] Non-limiting examples of haloalkyl also include difluoromethyl, 1,1,2,2-tetrafluoroethyl, perfluoroethyl, and the like.

[0084] In the present invention, haloalkoxy refers to an alkoxy group substituted by one or more halogens, wherein alkoxy is as defined above;

[0085] The halogenated alkoxy group may be fully halogenated or partially halogenated, and the number of halogenations may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.; the halogen is preferably F, Cl, Br, I; for example, it may be trifluoromethoxy, difluoromethoxy, 1,1,2,2-tetrafluoroethoxy, perfluoroethoxy, etc.

[0086] In the present invention, hydroxyalkyl refers to an alkyl group substituted by a hydroxy group, wherein the alkyl group is as defined above.

[0087] In the present invention, alkenyl refers to a chain alkenyl group, also known as an alkene group, preferably an alkenyl group containing 2 to 8 carbon atoms, more preferably an alkenyl group containing 2 to 6 carbon atoms, further preferably an alkenyl group containing 2 to 4 carbon atoms, and most preferably an alkenyl group containing 2 to 3 carbon atoms. Non-limiting examples of alkenyl groups include: ethenyl and propenyl. The alkenyl group may be further substituted with other related groups, for example: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate groups.

[0088] In the present invention, a haloalkyl group refers to an alkyl group substituted by one or more halogens, wherein the alkyl group is as defined above.

[0089] In the present invention, a haloalkoxy group refers to an alkoxy group substituted by one or more halogen groups, wherein the alkoxy group is as defined above.

[0090] In the present invention, hydroxyalkyl refers to an alkyl group substituted by a hydroxy group, wherein the alkyl group is as defined above.

[0091] In the present invention, a hydroxyl group refers to an -OH group.

[0092] In the present invention, halogen refers to fluorine, chlorine, bromine or iodine.

[0093] In the present invention, amino group refers to -NH2.

[0094] In the present invention, cyano refers to -CN.

[0095] In the present invention, the carboxyl group refers to -C(O)OH.

[0096] In the present invention, a carbonyl group refers to -C(O).

[0097] The hydrogen atoms described in the present invention can all be replaced by their isotope deuterium, and any hydrogen atom in the example compounds of the present invention can also be replaced by a deuterium atom.

[0098] In the present invention, "optional" or "optionally" means that the event or circumstances described subsequently may but need not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and this description includes both situations in which the heterocyclic group is substituted with an alkyl group and situations in which the heterocyclic group is not substituted with an alkyl group.

[0099] In the present invention, "substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms, in a group are independently replaced by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) whether substitution is possible or not without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when combined with a carbon atom with an unsaturated (e.g., olefinic) bond.

[0100] In the present invention, any substitution on an aliphatic ring, an aliphatic heterocycle, an aromatic ring, or an aromatic heterocycle means that one or more hydrogen atoms (preferably up to 5, more preferably 1 to 3 hydrogen atoms) at any substitutable position on the aliphatic ring, an aliphatic heterocycle, an aromatic ring, or an aromatic heterocycle are independently replaced by one or more substituents.

[0101] The compounds of this patent application include isotopic derivatives thereof. The term "isotopic derivative" refers to a compound whose structure differs only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of this patent application, with "deuterium" or "tritium" replacing hydrogen, or with 18F-fluorine labeling (18F isotope) replacing fluorine, or with 11C-, 13C-, or 14C-enriched carbon (11C-, 13C-, or 14C-carbon labeling; 11C-, 13C-, or 14C-isotope) replacing carbon atoms are within the scope of this patent application. Such compounds can be used as analytical tools or probes in, for example, biological assays, or can be used as in vivo diagnostic imaging tracers for diseases, or as tracers for pharmacodynamics, pharmacokinetics, or receptor studies. The various deuterated forms of the compounds of this patent application refer to compounds in which each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of compounds with reference to relevant literature. Deuterated forms of the compounds can be prepared using commercially available deuterated starting materials, or they can be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated iodoethane, and deuterated iodomethane. Deuterated compounds generally retain comparable activity to the undeuterated compounds, and when deuterated at certain sites, they can achieve improved metabolic stability, thereby conferring certain therapeutic advantages.

[0102] The compounds of this patent application may exist in specific stereoisomeric forms. The term "stereoisomer" refers to isomers with the same structure but different arrangements of atoms in space. It includes cis and trans (or Z and E) isomers, (-)- and (+)-isomers, (R)- and (S)-enantiomers, diastereomers, (D)- and (L)-isomers, tautomers, atropisomers, conformers and mixtures thereof (such as racemates, mixtures of diastereomers). The substituents in the compounds of this patent application may have additional asymmetric atoms. All of these stereoisomers and their mixtures are included within the scope of this patent application. Optically active (-)- and (+)-isomers, (R)- and (S)-enantiomers and (D)- and (L)-isomers can be prepared by chiral synthesis, chiral reagents or other conventional techniques. An isomer of a compound of the present patent application can be prepared by asymmetric synthesis or the use of a chiral auxiliary, or, when the molecule contains a basic functional group (e.g., an amino group) or an acidic functional group (e.g., a carboxyl group), by forming a diastereomeric salt with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art to obtain the pure isomer. Furthermore, separation of enantiomers and diastereoisomers is typically accomplished by chromatography.

[0103] In the chemical structure of the compound described in this patent application, the bond “ / ” indicates an unspecified configuration, that is, if chiral isomers exist in the chemical structure, the bond “ / ” can be or or contain both and configurations.

[0104] The compound of this patent application can exist in different tautomeric forms, and all such forms are included in the scope of this patent application.Term " tautomer " or " tautomeric form " refer to the structural isomer that exists in equilibrium and is easily converted into another isomeric form from one isomeric form.It includes all possible tautomers, i.e. exists in the form of a single isomer or in the form of a mixture of any proportions of the tautomer.Non-limiting examples include: keto-enol, imine-enamine, lactam-lactim etc.All tautomeric forms are within the scope of this patent application, and the naming of compound does not exclude any tautomer.

[0105] In the present invention, a pharmaceutical composition refers to a mixture containing one or more compounds described herein, or their physiologically / pharmaceutically acceptable salts or prodrugs, and other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredient and thereby exerting its biological activity.

[0106] In the present invention, pharmaceutically acceptable salts refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.

[0107] With respect to a drug or pharmacologically active agent, the term "therapeutically effective amount" refers to an amount of the drug or agent sufficient to achieve, or at least partially achieve, the desired effect. The determination of a therapeutically effective amount varies from person to person, depending on the age and general condition of the recipient, as well as the specific active substance. The appropriate therapeutically effective amount in each individual case can be determined by those skilled in the art through routine experimentation.

[0108] The term "pharmaceutically acceptable" as used herein refers to compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with patient tissues without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio, and effective for the intended use.

[0109] As used herein, the singular form "a," "an," and "the" include plural references and vice versa unless the context clearly dictates otherwise.

[0110] In the present invention, "plurality" refers to 2 or more, for example, it can be an integer such as 2, 3, 4, 5, 6, 7, 8, etc.

[0111] When the term "about" is applied to a parameter such as pH, concentration, temperature, etc., it indicates that the parameter can vary by ±10%, and sometimes more preferably within ±5%. As will be understood by those skilled in the art, when a parameter is not critical, numbers are generally given for illustration purposes only and are not limiting.

[0112] Specific implementation examples

[0113] The technical solution of the present invention is further described in detail below in conjunction with the specification and specific implementation examples, but the implementation of the present invention is not limited thereto. Equivalent replacements, combinations, improvements or modifications made by those skilled in the art to the technical solution of the present invention based on the description of the present invention should all be included in the scope of protection of the present invention.

[0114] Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the reagents and materials used in the following examples are all commercially available.

[0115] Example 1 Synthesis of Compound KH01

[0116]

[0117] Compound KH01-02: To a solution of compound KH01-01 (5 g, 31.53 mmol) and compound a (15.714 g, 37.84 mmol) in tetrahydrofuran (100 mL) at 0°C was added potassium tert-butoxide (5.307 g, 47.30 mmol) and allowed to react at room temperature for 2 hours. After completion, the reaction mixture was quenched with water (500 mL) and extracted with ethyl acetate (200 mL x 3). The organic phase was washed with saturated brine (300 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain a white solid (4.1 g, yield 60.6%). LCMS (ESI, m / z): 215.2 [MH] -1 . 1 H NMR (400MHz, DMSO) δ12.49(s,1H),7.53(d,J=7.8Hz,1H),7.37–7.27(m,2H),6.79(d,J=15.9Hz,1H),6.53–6.39(m,1H),3.29(dd,J=7.1,1.5Hz,2H).

[0118] Compound KH01-03: To a solution of compound KH01-02 (4 g, 18.64 mmol) in ethyl acetate (40 mL) was added palladium on carbon (600 mg). The mixture was reacted under a hydrogen atmosphere at room temperature for 1 h. After completion of the reaction, the reaction mixture was filtered and the filtrate was concentrated under vacuum to afford a white solid (2.5 g, 61.9% yield). 1 H NMR (400 MHz, DMSO-d6) δ 12.18 (s, 1H), 7.36–7.22 (m, 2H), 7.18 (d, J = 7.5 Hz, 1H), 2.84–2.68 (m, 2H), 2.26 (t, J = 7.3 Hz, 2H), 1.92–1.69 (m, 2H). Compound KH01-04: To a solution of compound KH01-03 (1.5 g, 6.92 mmol) in dichloromethane (20 mL) was added oxalyl chloride (1.2 mL, 13.85 mmol) and DMF (25 mg, 0.35 mmol) at 0°C. After stirring for 30 min, the system was concentrated under reduced pressure. A dichloromethane solution (20 mL) and aluminum chloride (1.385 g, 10.39 mmol) were added, and the reaction was carried out at 40°C for 1 h. After the reaction was complete, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain a light yellow solid (1.12 g, yield 81.5%). LCMS (ESI, m / z): 199.0 [M+H] +1 . 1 H NMR (400MHz, DMSO-d6) δ7.83(dd,J=8.4,5.8Hz,1H),7.32(dd,J=15.3,7.1Hz,1H),2.92–2.84(m,2H),2.42–2.35(m,2H),2.03–1.94(m,2H).

[0119] Compound KH01-05: To a solution of compound KH01-04 (1.12 g, 5.64 mmol) in ethanol (20 mL) were added hydroxylamine hydrochloride (980 mg, 14.10 mmol) and sodium acetate (1.156 g, 14.10 mmol), and the mixture was allowed to react at 90°C for 1 h. After the reaction was complete, the reaction solution was concentrated under vacuum to obtain a crude product, which was then dissolved in water and filtered. The filter cake was concentrated under vacuum to obtain a yellow solid (1.15 g, 95.4% yield). LCMS (ESI, m / z): 214.0 [M+H] +1 . 1H NMR (400MHz, DMSO-d6) δ11.28(s,1H),7.88(dd,J=8.8,5.7Hz,1H),7.25(t,J =8.9Hz,1H),2.80(t,J=6.0Hz,2H),2.64(t,J=6.5Hz,2H),1.84–1.74(m,2H).

[0120] Compound KH01-06: Polyphosphoric acid (10 mL) was added to compound KH01-05 (1.15 g, 5.38 mmol) and the mixture was allowed to react overnight at 80°C. After completion of the reaction, water (100 mL) was added to the reaction solution to quench the reaction. The mixture was then extracted with ethyl acetate (50 mL × 3). The organic phase was washed with saturated brine (50 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:1) to obtain a brown solid (520 mg, 45.2% yield). LCMS (ESI, m / z): 214.0 [M+H] +1 . 1 HNMR (400MHz, DMSO-d6) δ9.67(s,1H),7.29(t,J=8.9Hz,1H),6.99(dd,J=8.8,4.9Hz,1H),2.87(t,J=6.8Hz,2H),2.16–2.07(m,4H).

[0121] Compound KH01-07: To a solution of compound KH01-06 (500 mg, 2.34 mmol) in tetrahydrofuran (10 mL) at 0°C was added lithium aluminum tetrahydride (178 mg, 4.68 mmol), and the mixture was allowed to react at 80°C for 3 h. After completion of the reaction, water (0.2 mL) and 15% aqueous sodium hydroxide solution (0.2 mL) were added to the reaction mixture, and the mixture was stirred for 10 min. Water (0.6 mL) and anhydrous sodium sulfate were then added. The mixture was stirred for another 30 min, and the mixture was filtered. The filtrate was concentrated under vacuum to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain a yellow oil (390 mg, 83.5% yield). LCMS (ESI, m / z): 202.0 [M+H] +1 . 1 HNMR (400MHz, DMSO-d6) δ6.98(t,J=8.9Hz,1H),6.80(dd,J=8.6,5.1Hz,1H),5.37(s,1H),2.93–2.82(m,4H),1.71–1.64(m,2H),1.60–1.51(m,2H).

[0122] Compound KH01-08: Compound b (200 mg, 0.99 mmol) and T4P (6.5 g, 9.04 mmol) were added to a solution of compound KH01-07 (180 mg, 0.90 mmol) in pyridine (5 mL) and allowed to react overnight at room temperature. After completion, water (30 mL) was added to the reaction mixture to quench the reaction. The mixture was then extracted with ethyl acetate (20 mL × 3). The organic phase was washed with saturated brine (20 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 1:3) to obtain a yellow oil (100 mg, yield 29.0%). LCMS (ESI, m / z): 383.1 [M+H] +1 Compound KH01-09: To a mixture of compound KH01-08 (90 mg, 0.24 mmol), compound c (85 mg, 0.32 mmol), cesium carbonate (153 mg, 0.47 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (27 mg, 0.05 mmol), and tris(dibenzylideneacetone)dipalladium (22 mg, 0.02 mmol) was added dioxane (3 mL) at room temperature and reacted at 120°C under nitrogen for 1.5 hours. After the reaction was complete, the reaction solution was filtered, and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain a white solid compound (100 mg, yield 78.5%). LCMS (ESI, m / z): 542.2 [M+1] + .

[0123] Compound KH01: Dioxane hydrochloride (4M) (6 mL) was added to compound KH01-09 (100 mg, 0.18 mmol) at room temperature. The reaction was allowed to proceed overnight at room temperature and then at 60°C for 5.5 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was then purified by reverse phase preparative chromatography (FA) to obtain compound KH01 (26.13 mg, 28.2% yield). LCMS (ESI, m / z): 502.1 [M+1] + . 1H NMR(400MHz, DMSO-d6)δ8.23(d,J=13.9Hz,1H),7.69–7.49(m,1H),7.48–7.36(m,2H),5.77–5.68(m,1H),5.61–4.69(m,2H),4.53–3.98(m, 3H),3.82–3.40(m,1H),3.30–3.24(m,1H),3.23–2.98(m,1H),2.72–2 .53(m,3H),2.05–1.91(m,1H),1.79–1.58(m,2H),1.43–1.25(m,1H).

[0124] Example 2 Synthesis of Compound KH02 and Compound KH03

[0125]

[0126] Compound KH02-02: Triphenylmethylphosphonium iodide (9.1 g, 22.50 mmol) and potassium tert-butoxide (1.1 g, 22.50 mmol) were added to a tetrahydrofuran (50 mL) solution at 0°C. After stirring for 1 hour, compound 1 (5.0 g, 15.00 mmol) was added and allowed to react overnight at room temperature under hydrogen. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product, which was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain a white solid (2.7 g, 54.3% yield). LCMS (ESI, m / z): 332.1 [M+H] +1 .

[0127] Compound KH02-03: To a solution of compound KH02-02 (2.0 g, 6.03 mmol) in 1,2-dichloroethane (20 mL) at 0°C were added chloroiodomethane (4.4 mL, 60.35 mmol) and diethylzinc (1.0 M) (30 mL, 30.17 mmol) dropwise. The mixture was then allowed to react at 30°C for 2 days. After completion, the reaction mixture was quenched with saturated aqueous ammonium chloride and extracted with dichloromethane. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to afford a white solid (1.35 g, 64.8% yield). LCMS (ESI, m / z): (KH02-02) 332.1 [M+H] + 、(KH02-03)346.1[M+H] + .

[0128] Compound KH02-04 / KH03-04: Acetic acid (10 mL), hydrochloric acid (10 mL), and water (5 mL) were added to a mixture of compounds KH02-03 / KH02-02 (1.35 g) at room temperature and allowed to react at 120°C for 3 days. After completion of the reaction, the pH was adjusted to neutral by adding saturated aqueous sodium bicarbonate solution. The mixture was then extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain the crude product, which was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 5:1) to obtain a yellow oil (350 mg, 46.8% yield). LCMS (ESI, m / z): (KH02-04) 192.1 [M+H] + 、(KH03-04)178.1[M+H] + .

[0129] Compound KH02-05 / KH03-05: Compound b (498 mg, 3.73 mmol) was added to a solution of compound a (250 mg, 1.24 mmol) in dichloromethane (10 mL) at 0°C. After stirring for 0.5 hours, a mixture of compounds KH02-04 / KH03-04 (309 mg) and pyridine (393 mg, 4.97 mmol) were added and the mixture was allowed to react at 30°C overnight. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to obtain a crude product. The crude product was then purified by silica gel column chromatography (ethyl acetate) to obtain a white solid compound (180 mg, 38.7% yield). LCMS (ESI, m / z): (KH03-05) 361.2 [M+1] + 、(KH02-05)375.2[M+1] + .

[0130] Compound KH02-06 / KH03-06: To a mixture of compound KH02-05 / KH03-05 (180 mg), compound c (173 mg, 0.72 mmol), cesium carbonate (313 mg, 0.96 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (56 mg, 0.10 mmol), and tris(dibenzylideneacetone)dipalladium (44 mg, 0.05 mmol) was added dioxane (5 mL) and reacted at 120°C under nitrogen for 2 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain the compound as a white solid (170 mg, yield 66.3%). LCMS (ESI, m / z): 519.9 [M+1] + .

[0131] Compounds KH02 and KH03: To the KH02-06 / KH03-06 mixture (150 mg) was added dioxane hydrochloride (4 M) (5 mL) at room temperature and allowed to react at 60°C for 4 hours. After the reaction was complete, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was then purified by reverse-phase preparative chromatography (NH3.H2O) to obtain compounds KH02 (10.55 mg) and KH03 (38.31 mg). KH02: LCMS (ESI, m / z): 494.1 [M+1] + . 1 HNMR(400MHz, DMSO-d6)δ8.21(d,J=7.5Hz,1H),7.93–7.54(m,1H),7.46–7.28(m,3H),6.08–5.93(m,1H),5.76–5.66(m,1H),5.13–4.63(m, 1H),4.53–4.16(m,3H),4.10–3.57(m,1H),2.69–2.57(m,3H),2.46–2 .42(m,1H),2.35–2.17(m,2H),2.02–1.92(m,1H),1.27–0.89(m,4H).

[0132] KH03: LCMS (ESI, m / z): 480.0[M+1] + . 1 H NMR(400MHz,DMSO-d6)δ8.18(s,1H),7.87–7.51(m,1H),7.47–7.22(m,3H),6.13– 5.94(m,1H),5.76–5.62(m,1H),5.50–5.04(m,1H),4.80–4.53(m,1H),4.50–4.34( m, 1H), 4.29–4.16 (m, 1H), 3.71–3.42 (m, 1H), 3.29–3.06 (m, 1H), 2.61 (s, 2H), 2.49–2.39 (m, 2H), 2.35–2.25 (m, 1H), 2.21 (s, 1H), 2.17–2.07 (m, 2H). Example 3 Synthesis of Compound KH04

[0133]

[0134] Compound KH04-02: Palladium on carbon (10%) (321 mg, 0.30 mmol) was added to a solution of compound 1 (1.0 g, 3.02 mmol) in tetrahydrofuran (10 mL) and methanol (20 mL) at room temperature. The mixture was reacted under hydrogen at room temperature for 1 h. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated under vacuum to obtain a white solid (1.0 g, 99.4% yield). LCMS (ESI, m / z): 334.1 [M+H] + Compound KH04-03: Polyphosphoric acid (5 mL) was added to compound KH04-02 (1.0 g, 3.02 mmol) and the mixture was allowed to react at 120°C for 2 h. After the reaction was complete, water was added to the reaction solution, the pH was adjusted to neutral with potassium carbonate, and ethyl acetate was added for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to obtain a crude product, which was then purified by silica gel column chromatography (petroleum ether: ethyl acetate = 10:1) to obtain a colorless oil (50%) (600 mg, yield 55.8%). LCMS (ESI, m / z): 180.1 [M+H] + .

[0135] Compound KH04-04: Compound a (240 mg, 1.19 mmol), compound KH04-03 (321 mg, 1.79 mmol), and butylphosphonic anhydride (50%) (8.6 g, 11.93 mmol) were dissolved in pyridine (10 mL) at room temperature and reacted under nitrogen overnight. After the reaction was complete, water was added to the reaction solution and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was then purified by silica gel column chromatography (ethyl acetate) to obtain a white solid compound (220 mg, 50.9% yield). LCMS (ESI, m / z): 363.1 [M+1] + .

[0136] Compound KH04-05: To a mixture of compound KH04-04 (220 mg, 0.61 mmol), compound c (219 mg, 0.91 mmol), cesium carbonate (396 mg, 1.21 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (70 mg, 0.12 mmol), and tris(dibenzylideneacetone)dipalladium (56 mg, 0.06 mmol) was added dioxane (5 mL) and reacted at 120°C under nitrogen for 2 hours. After the reaction was complete, the reaction solution was filtered and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain the compound as a white solid (150 mg, 47.4% yield). LCMS (ESI, m / z): 522.2 [M+1] + .

[0137] Compound KH04: Dioxane hydrochloride (4M) (5 mL) was added to compound KH04-05 (150 mg, 0.29 mmol) at room temperature. The reaction was allowed to proceed overnight at room temperature and then at 60°C for 5 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was then purified by reverse phase preparative chromatography (FA) to obtain compound KH04 (25.68 mg, 18.5% yield). LCMS (ESI, m / z): 482.3 [M+1]. + . 1 H NMR(400MHz, DMSO-d6)δ8.22(d,J=14.7Hz,1H),7.74–7.45(m,1H),7.43–7.39(m,1H),7. 26–7.19(m,1H),7.18–7.02(m,1H),5.87–5.36(m,2H),5.25–5.18(m,1H),4.80–4.35(m, 1H),4.33–4.27(m,1H),4.25–4.08(m,1H),3.84–3.55(m,1H),2.69–2.53(m,3H),2.47–2 .29(m,1H),1.84–1.72(m,2H),1.59–1.45(m,1H),1.36–1.28(m,3H),1.25–1.02(m,1H).

[0138] Example 4 Synthesis of Compound KH05

[0139]

[0140] Compound KH05-02: To a solution of compound KH05-01 (4.0 g, 24.36 mmol) in ethanol (50 mL) were added hydroxylamine hydrochloride (4.23 g, 60.91 mmol) and sodium acetate (5.0 g, 60.91 mmol) at room temperature and reacted at 90°C for 2 hours. After the reaction was complete, the reaction solution was concentrated, water was added, and filtration afforded the compound as a white solid (4.2 g, 96.2% yield). LCMS (ESI, m / z): 180.2 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δ11.08(s,1H),7.92–7.82(m,1H),7.09–6.95(m,2H),2.76–2.68(m,2H),2.64(t,J=6.6Hz,2H),1.83–1.69(m,2H).

[0141] Compound KH05-03: Polyphosphoric acid (40 mL) was added to compound KH05-02 (4.2 g, 23.44 mmol) at room temperature and allowed to react overnight at 80°C. After completion of the reaction, ethyl acetate and water were added to the reaction solution for extraction. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a brown solid compound (2.1 g, 50.0% yield). LCMS (ESI, m / z): 180.2 [M+1] + . 1 H NMR (400MHz, DMSO-d6) δ9.48(s,1H),7.14(dd,J=9.3,2.9Hz,1H),7.09–7.01(m,1H),7.01–6.95(m,1H),2.68(t,J=6.8Hz,2H),2.19–2.04(m,4H).

[0142] Compound KH05-04: Tetrahydrofuran (10 mL) was added to lithium aluminum tetrahydride (424 mg, 11.16 mmol) at room temperature, followed by compound KH05-03 (1.0 g, 5.58 mmol) at 0°C. The mixture was then reacted at 80°C for 5 hours. After completion of the reaction, tetrahydrofuran was added to dilute the mixture. 0.5 mL of water, 0.5 mL of 15% aqueous NaOH solution, and 1.5 mL of water were added sequentially. The mixture was stirred for 10 minutes and filtered. The filtrate was concentrated under vacuum to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 10:1) to obtain the compound as a white solid (760 mg, 82.4% yield). LCMS (ESI, m / z): 166.3 [M+1] + . 1 H NMR(400MHz,DMSO-d6)δ6.92–6.84(m,1H),6.84–6.71(m,2H),5.15(s,1H), 2.92–2.80(m,2H),2.73–2.57(m,2H),1.71–1.61(m,2H),1.57–1.47(m,2H).

[0143] Compound KH05-05: Compound a (300 mg, 1.49 mmol), compound KH05-04 (493 mg, 2.98 mmol), and T3P (9.5 g, 14.91 mmol) were dissolved in pyridine (10 mL) at room temperature under nitrogen and allowed to react overnight. After the reaction was complete, water was added to the reaction solution and extracted with ethyl acetate. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product. The crude product was then purified by silica gel column chromatography (ethyl acetate) to obtain a white solid compound (300 mg, yield 57.8%). LCMS (ESI, m / z): 349.2 [M+1]+ . 1 HNMR(400MHz,DMSO-d6)δ7.97(d,J=17.3Hz,1H),7.48–7.31(m,1H),7.29–7.05(m,2H),4.64–4.31(m,3H),3.75–3.5 9(m,1H),3.17(t,J=13.1Hz,1H),2.79–2.53(m,3H),1.97–1.85(m,1H),1.83–1.62(m,2H),1.32(s,3H),1.26(s,3H).

[0144] Compound KH05-06: To a mixture of compound KH05-05 (300 mg, 0.86 mmol), compound c (310 mg, 1.29 mmol), cesium carbonate (561 mg, 1.72 mmol), Xantphos (100 mg, 0.17 mmol), and Pd2(dba)3 (79 mg, 0.09 mmol) was added dioxane (5 mL) at room temperature and allowed to react at 120°C under nitrogen for 2 hours. After completion of the reaction, the reaction solution was filtered and the filtrate was concentrated under vacuum to obtain the crude product. The crude product was then purified by silica gel column chromatography (petroleum ether:ethyl acetate = 3:1) to obtain the compound as a white solid (400 mg, 91.5% yield). LCMS (ESI, m / z): 508.2 [M+1] + .

[0145] Compound KH05: Dioxane hydrochloride (4M) (4 mL) was added to compound KH05-06 (200 mg, 0.39 mmol) at room temperature. The reaction was allowed to proceed overnight at room temperature and then at 60°C for 3.5 hours. After completion of the reaction, the reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was then purified by reverse-phase preparative chromatography (formic acid / acetonitrile) to obtain compound KH05 (63.01 mg, 34.2% yield). LCMS (ESI, m / z): 468.2 [M+1] + . 1H NMR(400MHz, DMSO)δ8.22(d,J=14.5Hz,1H),7.67–7.49(m,1H),7.42(d,J=6.9Hz,1H),7 .35–7.13(m,2H),5.74–5.65(m,1H),5.57–5.21(m,1H),4.70–4.45(m,1H),4.38–4.12(m ,2H),3.72–3.67(m,1H),3.51(t,J=13.3Hz,1H),3.22–3.09(m,1H),2.84–2.70(m,1H),2 .59(s,3H),2.54–2.52(m,1H),2.02–1.88(m,1H),1.76–1.60(m,2H),1.42–1.21(m,1H).

[0146] Example 5 In vitro activity detection of compounds

[0147] 5.1 Experimental reagents and consumables

[0148] Table 1

[0149]

[0150] 5.2 Experimental steps

[0151] Compounds were serially diluted in DMSO using a 384PP Plate. Using an Echo, 0.15 μL of compound was transferred to a 384-reaction microplate (Greiner 784075), ensuring a final DMSO content of 1% (in duplicate). 5 μL of 3X POLQ-C enzyme solution was added to each well of the 384-reaction microplate and incubated at 25°C for 10 minutes. The well containing DMSO and POLQ-C enzyme served as a high control, while the well containing DMSO and assay buffer served as a low control. 5 μL of 3X FITC-dATP, dsDNA, and a mixed solution of dCTP, dGTP, and dTTP were added to each well. Finally, 5 μL of Streptavidin-Tb cryptate solution was added to each well and incubated at 25°C for 60 minutes. (Final concentrations: 5 nM POLQ-C, 10 nM dsDNA, 10 μM dCTP & dGTP & dTTP, 0.1 μM FITC-dATP, 1X Streptavidin-Tb cryptate). The signals at 490 nm and 520 nm were read on a BMG (PHERAstar FSX) microplate reader, and the ratio was calculated (Ratio: 520 / 490*10 4The percentage inhibition of compound-treated wells was normalized between High Control and Low Control (%inhibition = (AVE High Control -Ratio 化合物读值 ) / (AVE High Control -AVE Low Control )*100). Then, the four-parameter IC was fitted by XLfit 5.5.0. 50 Curve and analysis, IC 50 The IC50 of the compounds described in the present invention is 1-100 nM. In particular, the IC50 of compound KH01 reaches 4 nM.

Claims

1. A compound of the following formula, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that The compound structure is as follows: wherein R1 and R2 are each independently selected from H, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, and heteroaryl; R3 is selected from H, amino, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl; or said R3 and R1 or R2 and adjacent N and C form a heterocyclic group, said heterocyclic group is optionally substituted by hydrogen, halogen, amino, hydroxy, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy; R4 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, carbonyl, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclic group, aryl, heteroaryl; R5 is each independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl; B is aryl or heteroaryl, said aryl or heteroaryl being optionally substituted with hydrogen, halogen, amino, hydroxy, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy; n is an integer from 0 to 4.

2. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that R3 is an aryl group or a heteroaryl group, wherein the heteroaryl group contains at least one atom selected from N, O, and S.

3. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that The compound has the structure shown in Formula II: wherein A1, A2, A3, and A4 are each independently selected from CR6, CR6R6a, NR6, O, S, and S(O)2; and R6 is independently selected from hydrogen, hydroxy, halogen, alkyl, haloalkyl, alkoxy, amino, aminoalkyl, acyl, cycloalkyl, aryl, heterocyclyl, and heteroaryl; or A1 and A2, A2 and A3 optionally form a substituted or unsubstituted saturated or unsaturated 3-14 membered ring, which may optionally include one or more heteroatoms, which may be the same or different and are independently selected from O, N, and S; R6a is independently selected from hydrogen, hydroxy, halogen, alkyl, haloalkyl, alkoxy, amino, aminoalkyl, acyl; or R6 and R6a together with the carbon atom to which they are attached form a -C(=S)-, (-C)=O, -C(=NH)- group, or a substituted or unsubstituted saturated or unsaturated 3-14 membered ring which may optionally include one or more heteroatoms which may be the same or different and which are independently selected from O, N, and S; B is aryl or heteroaryl, said aryl or heteroaryl being optionally substituted with hydrogen, hydroxy, halogen, haloalkyl, alkoxy, amino, aminoalkyl, acyl, alkyl, cycloalkyl, aryl, heterocyclyl and heteroaryl; R4 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, carbonyl, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl; R5 is each independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl.

4. The compound according to claim 3, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that The compound has a structure as shown in Formula III: in: A1 is selected from CR6, (-C)=O, S(O)2, R6 is independently selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl; A2 is selected from CR6, NR6, O, R6 is independently selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl; Or A1 and A2 form a 5-6 membered aryl or heteroaryl group; R7 is selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl; The other substituents are as defined in claim 4.

5. The compound according to claim 3, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that: Selected from:

6. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that: B is selected from a 5-6 membered aryl or heteroaryl group, wherein the heteroaryl group may optionally include one or more heteroatoms, which may be the same or different and are independently selected from O, N, and S, and the aryl or heteroaryl group may be optionally substituted with hydrogen, hydroxy, halogen, cyano, alkyl, haloalkyl, alkoxy, amino, or aminoalkyl; Preferably, B is selected from: in: A5, A6, A7, A8 are CR11 or NR11, R11 is independently selected from hydrogen, hydroxy, halogen, substituted or unsubstituted alkyl; R8 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl; More preferably, B is selected from: R8 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl.

7. The compound according to claim 6, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that The compound has a structure as shown in Formula IV: R8 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl R4 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, carbonyl, alkyl, alkenyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl; R5 is independently selected from hydrogen, hydroxy, halogen, amino, cyano, carboxyl, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl.

8. The compound according to claim 7, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that: R8 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl.

9. The compound according to claim 7, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that: R4 is selected from hydrogen, hydroxy, halogen, amino, carbonyl, C1-6 alkyl, C2-6 alkenyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-6 cycloalkyl, C3-6 heterocyclyl; preferably, R4 is hydrogen; Preferably, R5 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl.

10. The compound according to claim 7, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that The compound has a structure as shown in Formula V: R8 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl. R9 and R10 are each independently selected from hydrogen, hydroxyl, halogen, amino, carbonyl, C1-6 alkyl, C2-6 alkenyl, C1-6 haloalkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-6 cycloalkyl, C3-6 heterocyclyl, wherein the C3-6 heterocyclyl may optionally include one or more O, N and S heteroatoms; preferably, R9 and R10 are hydrogen; R5 is selected from hydrogen, halogen, C1-6 alkyl, C1-6 haloalkyl.

11. The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that Has the following structure:

12. A pharmaceutical composition comprising a therapeutically effective dose of the compound according to any one of claims 1 to 11, its stereoisomers or pharmaceutically acceptable salts thereof and one or more pharmaceutically acceptable carriers or excipients.

13. Use of the compound according to any one of claims 1 to 11, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 12 in the preparation of a medicament for treating a Polθ-mediated disease; preferably, the Polθ-mediated disease is liver cancer, breast cancer, ovarian cancer, lung cancer, kidney cancer, prostate cancer, skin cancer, bladder cancer, pancreatic cancer or head and neck cancer.

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