Aryl alkyne compound and use thereof

Abstract

Disclosed in the present invention are an aryl alkyne compound and use thereof. Provided in the present invention is a compound represented by formula I or a pharmaceutically acceptable salt thereof. The compound provided in the present invention has a good inhibitory effect on the PARP1 enzyme.

Description

An aryl yne compound and its applications

[0001] This application claims priority to Chinese patent application 2025103674470, filed on March 26, 2025. The entire contents of the aforementioned Chinese patent application are incorporated herein by reference. Technical Field

[0002] This invention relates to an aryl alkyne compound and its applications. Background Technology

[0003] PARPs are a family of multifunctional post-translational modifying enzymes, with 17 family members (PARP1, PARP2, ... PARP17) identified to participate in various biological functions, including replication, recombination, chromosome remodeling, and DNA damage repair. Homologous recombination-deficient (HRD) tumor cells, especially BRCA1 / 2-deficient tumor cells, exhibit a combined lethal effect against PARP inhibitors (PARPi). PARPi, as a targeted cancer therapy, has been extensively studied, and more than six PARPis are currently on the market, such as Olaparib, Rucaparib, Niraparib, Talazoparib, Fluazoparib, and Pamiparib. However, PARPi resistance is very common clinically; over 40% of BRCA1 / 2-deficient patients do not respond to PARPi or develop resistance after treatment. Furthermore, PARPis all exhibit hematologic toxicity, affecting the duration and dosage of treatment, necessitating hematological monitoring of patients and timely management of any adverse events.

[0004] PARP1 and PARP2 are the most well-studied members of the PARP family, and all marketed PARP1 inhibitors are PARP1 / 2 inhibitors. PARP1 is activated at DNA damage sites, catalyzing the PARylation (poly ADP-ribosylation) reaction on target proteins, mediating the recruitment of DNA damage repair proteins, and is a key protein in the base excision repair pathway for repairing single-strand breaks (SSBs). The repair process is accompanied by the release of PARP1 / 2 and DNA. PARP1 gene deletion and BRCA1 / BRCA2-deficient tumor cells have a combined lethal effect. Studies have reported that PARP2 is an essential protein for the survival of hematopoietic stem cells / progenitor cells; PARP2 deficiency leads to a shortened erythrocyte lifespan, affects erythrocyte progenitor cell differentiation, and ultimately leads to hematologic toxicity. Known evidence suggests that PARP1 inhibition is the main source of PARP1 inhibitor tumor-suppressive activity, while PARP2 inhibition may be the source of hematologic toxicity. Therefore, this patent designs a selective inhibitor targeting PARP1.

[0005] Since HRD (especially BRCA1,2 mutations) is a prerequisite for the antitumor activity of PARPi, resistance eventually arises due to various factors leading to the restoration of homologous recombination repair, including DNA replication fork protection, excessive activation of parylation, reversion mutations, epigenetic alterations, and pharmacokinetic changes. Currently, combination therapy is mainly used to overcome PARPi resistance.

[0006] Developing next-generation PARPis, expanding their indications, and reducing their toxic side effects have always been key directions in new drug development. Summary of the Invention

[0007] The technical problem to be solved by this invention is to provide a novel small molecule inhibitor targeting PARP1. To this end, this invention provides an aryl alkyne compound and its applications. The alkyne-containing compound provided by this invention exhibits good inhibitory activity against PARP1.

[0008] This invention provides a compound as shown in Formula I or a pharmaceutically acceptable salt thereof.

[0009] L 1 For not replaced or by one or more R a Replacement C 2-14 alkynyl group;

[0010] Each R aIndependently unsubstituted or by one or more R a1 Substituted 6-10 aryl groups;

[0011] Each R a1 Independently, C is either unsubstituted or substituted with one or more hydroxyl groups. 1-6 alkyl;

[0012] L 2 For not replaced or by one or more R P2 Substituted 6-10 arylene groups or unsubstituted or substituted with one or more R groups P2 Substituted 5-10 heteroaryl groups;

[0013] L 3 For not replaced or by one or more R P2 Substituted 3-12 membered heterocyclic alkyl groups;

[0014] L 4 For not replaced or by one or more R P2 Replacement C 1-14 Alkylene;

[0015] Each R P2 Halogens are independent of each other;

[0016] Ring E is unsubstituted or replaced by one or more R e-1 Substituted 6-10 aryl rings or unsubstituted or substituted with one or more R e-1 Substituted 5-12 heterocyclic aromatic rings;

[0017] Ring F is unsubstituted or replaced by one or more R e-2 Substituted 4-12 membered olefinic heterocycles;

[0018] Each R e-1 Independent of halogen or unsubstituted C 1-6 alkyl;

[0019] Each R e-2 Independently an oxo group (=O), a halogen, or an unsubstituted C 1-6 alkyl;

[0020] The heteroatoms in the 5-10-membered heteroaryl, 3-12-membered heterocyclic alkyl, 5-12-membered heteroaromatic and 4-12-membered olefinic heterocycles are independently selected from one, two or three of N, O and S, and the number of heteroatoms is independently 1, 2, 3, 4 or 5; the number of olefin bonds in the 4-12-membered olefinic heterocycle is 1 or 2.

[0021] In a preferred embodiment, certain groups in the compound of Formula I or its pharmaceutically acceptable salt are defined as follows, and the definitions of groups not mentioned are as described in any embodiment of the present invention (hereinafter referred to as "in a preferred embodiment").

[0022] In a certain preferred solution, L 1 In, the unsubstituted or unreplaced R a Replacement C 2-14 C in the alkynyl group 2-14 The alkynyl group is independently C 2-6 alkynyl group, the C 2-6 The alkynyl group can be a straight-chain alkynyl group, for example... For example

[0023] In a certain preferred solution, R a In, the unsubstituted or unreplaced R a1 The 6-10 aryl group in the substituted 6-10 aryl group is phenyl or naphthyl, such as phenyl.

[0024] In a certain preferred solution, R a 1 In, the unsubstituted or unreplaced R a1-1 Replacement C 1-6 C in alkyl 1-6 The alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl, such as methyl.

[0025] In a certain preferred solution, L 2 In, the unsubstituted or unreplaced R P2 The 6-10 arylene group in the substituted 6-10 arylene group is independently phenylene or naphthylene, for example, phenylene, for example

[0026] In a certain preferred solution, L 2 In, the unsubstituted or unreplaced R P2 The 5-10-membered heteroaryl group in the substituted 5-10-membered heteroaryl group is independently a 5-6-membered heteroaryl group.

[0027] In a certain preferred solution, L 2 In, the unsubstituted or unreplaced R P2 The heteroatom in the substituted 5-10-membered heteroaryl group is N, and the number of heteroatoms can be 1, 2 or 3 independently.

[0028] In a certain preferred solution, L 2 In, the unsubstituted or unreplaced RP2 The 5-10-membered heteroaryl group in the substituted 5-10-membered heteroaryl group is independently a 5-6-membered heteroaryl group, wherein the heteroatom of the 5-6-membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2, or 3, such as pyridyl, pyrimidinyl, pyrazinyl, or pyridazinyl, for example...

[0029] In a certain preferred solution, L 2 In, the one or more R P2 The 5-10 methyl methacrylate is replaced by

[0030] In a certain preferred solution, L 4 In, the one or more R P2 The 5-10 aryl group that is replaced is

[0031] In a certain preferred solution, L 3 In, the unsubstituted or unreplaced R P2 The 3-12-membered heterocyclic alkyl group in the substituted 3-12-membered heterocyclic alkyl group is independently a monocyclic ring.

[0032] In a certain preferred solution, L 3 In, the unsubstituted or unreplaced R P2 The 3-12-membered heterocyclic alkyl group in the substituted 3-12-membered heterocyclic alkyl group is independently a 3-8-membered heterocyclic alkyl group, such as a 3-6-membered heterocyclic alkyl group.

[0033] In a certain preferred solution, L 3 In, the unsubstituted or unreplaced R P2 The heteroatoms of the substituted 3-12-membered heterocyclic alkyl group are independently selected from one or both of N and O, and the number of heteroatoms can be independently 1 or 2, for example, the heteroatom is N, and the number of heteroatoms is 1 or 2.

[0034] In a certain preferred solution, L 3 In, the unsubstituted or unreplaced R P2 The 3-12-membered heterocyclic alkyl group in the substituted 3-12-membered heterocyclic alkyl group is independently a 3-6-membered monocyclic heterocyclic alkyl group, wherein the heteroatom of the 3-6-membered monocyclic heterocyclic alkyl group is independently selected from one or both of N and O, and the number of heteroatoms is independently one or two; for example, piperazine group, and another example...

[0035] In a certain preferred solution, L 4 In, the unsubstituted or unreplaced R P2 Replacement C1-14 C in alkylene 1-14 Alkylene is independently C 1-8 Alkylene, the C 1-8 Alkylenes can be straight-chain alkylenes, for example For example

[0036] In a preferred embodiment, in ring E, the unsubstituted or unreplaced element is one or more R elements. e-1 The 6-10 membered aromatic ring of the substituted 6-10 membered aromatic ring is a benzene ring or a naphthalene ring, for example, a benzene ring.

[0037] In a preferred embodiment, in ring E, the unsubstituted or unreplaced element is one or more R elements. e-1 The substituted 5-12-membered heteroaromatic ring is a 5-10-membered heteroaromatic ring, such as a 5-6-membered heteroaromatic ring, such as a pyridine ring.

[0038] In a preferred embodiment, in ring E, the heteroatom of each of the 5-12 membered heteroaromatic rings is N, and the number of heteroatoms can be 1, 2 or 3.

[0039] In a preferred embodiment, in ring F, the unsubstituted or unreplaced element is one or more R elements. e-2 The 4-12 substituted olefinic heterocycles are independently 5-6 olefinic heterocycles.

[0040] In a preferred embodiment, in ring F, the unsubstituted or unreplaced element is one or more R elements. e-2 The heteroatom in the substituted 4-12-membered alkene heterocycle is N, and the number of heteroatoms can be 1 or 2.

[0041] In a preferred embodiment, in ring F, the unsubstituted or unreplaced element is one or more R elements. e-2 The number of alkene bonds in the substituted 4-12 alkene heterocycle is one or two.

[0042] In a preferred embodiment, in ring F, the unsubstituted or unreplaced element is one or more R elements. e-2 The 4-12-membered olefinic heterocycle is replaced with a 5-6-membered olefinic heterocycle, wherein the heteroatom of the 5-6-membered olefinic heterocycle is N, the number of heteroatoms is 1 or 2, and the number of olefin bonds is 1 or 2, for example...

[0043] In a preferred embodiment, in ring F, the element being controlled by one or more R... e-2 The substituted 4-12 membered olefinic heterocycle is

[0044] In a certain preferred solution, each R P2R e-1 and R e-2 In this context, the halogen is independently fluorine, chlorine, bromine, or iodine, such as fluorine.

[0045] In a certain preferred solution, each R e-1 and R e-2 In, the unsubstituted C 1-6 The alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, such as methyl or ethyl.

[0046] In a preferred embodiment, each of the "multiples" is two or three, for example, two.

[0047] In a certain preferred solution, L 1 For not replaced or by an R a Replacement C 2-6 Alkyne group.

[0048] In a certain preferred solution, each R a Independently for being a R a1 Replaced 6-10 aryl groups.

[0049] In a certain preferred solution, each R a1 Independently for C substituted with one hydroxyl group 1-6 alkyl.

[0050] In a certain preferred solution, L 2 For not replaced or by one or more R P2 Substituted phenylene or unsubstituted or with one or more R P2 The substituted 5-6-membered heteroaryl group; the heteroatom of the 5-6-membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2, or 3; preferably, L 2 It is an unsubstituted phenylene or an unsubstituted 5-6 membered heteroaryl group; the heteroatom of the 5-6 membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2 or 3.

[0051] In a certain preferred solution, L 3 For not replaced or by one or more R P2 The substituted 3-6-membered monocyclic heterocyclic alkylene group, wherein the heteroatoms of the 3-6-membered monocyclic heterocyclic alkylene group are independently selected from one or both of N and O, and the number of heteroatoms is independently one or two; preferably, L 3 It is an unsubstituted 3-6 membered monocyclic heterocyclic alkylene group, wherein the heteroatom of the 3-6 membered monocyclic heterocyclic alkylene group is N, and the number of heteroatoms is 1 or 2.

[0052] In a certain preferred solution, L 4 For not replaced or by one or more R P2Replacement C 1-8 Alkylene; preferably, L 4 For unreplaced C 1-8 Alkylene.

[0053] In a preferred embodiment, ring E is unsubstituted or substituted by one or more R. e-1 Substituted benzene ring or unsubstituted or with one or more R e-1 The substituted 5-6 membered heteroaromatic ring, wherein the heteroatom in the 5-6 membered heteroaromatic ring is N, and the number of heteroatoms is 1 or 2.

[0054] In a certain preferred solution, each R e-1 It is a halogen on its own.

[0055] In a preferred embodiment, ring F is unsubstituted or substituted by one or more R. e-2 The substituted 5-6 membered olefinic heterocycle, wherein the heteroatom of the 5-6 membered olefinic heterocycle is N, the number of heteroatoms is one or two, and the number of olefin bonds is one or two; preferably, the ring F is substituted by one or more R e-2 Substituted 5-6 membered olefinic heterocycles.

[0056] In a certain preferred solution, each R e-2 Independent of an oxo group or an unsubstituted C 1-6 alkyl.

[0057] In a certain preferred solution, L 1 for

[0058] In a certain preferred solution, L 2 for The better location is Better, for

[0059] In a certain preferred solution, L 2 for 1 bit and L 2 Connect, 2 bits and L 3 Connection; preferably, for 1 bit and L 2 Connect, 2 bits and L 3 Connect; better yet, for 1 bit and L 2 Connect, 2 bits and L 3 connect.

[0060] In a certain preferred solution, L3 for

[0061] In a certain preferred solution, L 4 for

[0062] In a certain preferred solution, for The better location is

[0063] In a certain preferred solution, L 1 For not replaced or by an R a Replacement C 2-6 alkynyl group;

[0064] R a For being an R a1 Substituted 6-10 aryl groups;

[0065] R a1 C replaced by a hydroxyl group 1-6 alkyl;

[0066] L 2 For not replaced or by one or more R P2 Substituted phenylene or unsubstituted or with one or more R P2 The substituted 5-6-membered heteroaryl group; wherein the heteroatom of the 5-6-membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2 or 3;

[0067] L 3 It is an unsubstituted 3-6 membered monocyclic heterocyclic alkylene group, wherein the heteroatom of the 3-6 membered monocyclic heterocyclic alkylene group is N, and the number of heteroatoms is 1 or 2;

[0068] L 4 For unreplaced C 1-8 Alkylene;

[0069] Ring E is unsubstituted or replaced by one or more R e-1 Substituted benzene ring or unsubstituted or with one or more R e-1 The substituted 5-6 membered heteroaromatic ring, wherein the heteroatom in the 5-6 membered heteroaromatic ring is N, and the number of heteroatoms is 1 or 2;

[0070] Each R e-1 Halogens are independent of each other;

[0071] Ring F is formed by one or more R e-2 A substituted 5-6 membered olefinic heterocycle; wherein the heteroatom of the 5-6 membered olefinic heterocycle is N, the number of heteroatoms is 1 or 2, and the number of olefin bonds is 1 or 2;

[0072] Each R e-2 Independent of an oxo group or an unsubstituted C 1-6 alkyl.

[0073] In a preferred embodiment, the compound shown in Formula I is any of the following compounds:

[0074] This invention provides a pharmaceutical composition comprising:

[0075] (1) (Therapeuticly effective amount) of the compound of formula I as described in any one of the present invention, or a pharmaceutically acceptable salt thereof, and

[0076] (2) Pharmaceutical excipients.

[0077] This invention provides the use of substance A in the preparation of PARP1 enzyme inhibitors, wherein substance A is a compound of formula I as described in any one of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0078] This invention provides the use of substance A in the preparation of medicaments for treating and / or preventing diseases associated with the PARP1 enzyme, such as breast cancer, colorectal cancer, prostate cancer, or pancreatic cancer, wherein substance A is a compound of formula I as described in any one of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0079] This invention provides the use of substance A in the preparation of medicaments for treating and / or preventing breast cancer, colorectal cancer, prostate cancer, or pancreatic cancer, wherein substance A is a compound of formula I as described in any one of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0080] The present invention provides a substance A for use as a treatment, said substance A being a compound of formula I as described in any one of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0081] The present invention provides a substance A for treating and / or preventing breast cancer, colorectal cancer, prostate cancer, or pancreatic cancer, wherein substance A is a compound of formula I as described in any one of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0082] The present invention provides a method for treating and / or preventing breast cancer, colorectal cancer, prostate cancer, or pancreatic cancer, comprising the steps of: administering to a subject a therapeutically effective amount of substance A, said substance A being a compound of Formula I as described in any one of the present invention, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0083] Detailed explanation: Unless otherwise stated, the terms used in the specification and claims shall have the following meanings.

[0084] Those skilled in the art will understand that, according to conventions used in the art, the structural formulas of the groups described in this invention are... This refers to the fact that the corresponding group is connected to other fragments or groups in the compound through this site.

[0085] In this article, a single hyphen "-" can be added before the substituents used to indicate that the named substituent is connected to the parent part by a single bond.

[0086] When any variable (e.g., R) a When a group appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is surrounded by 0-2 R... a If replaced, the group may optionally be replaced by at most two R groups. a Replaced, and R in each case a Each has its own independent options. Furthermore, combinations of substituents and / or their variants are only permitted if such combinations produce stable compounds.

[0087] The term "multiple" refers to 2, 3, 4 or 5, preferably 2 or 3.

[0088] In the claims of this application, "one or more" in "satisfying one or more of the following conditions" means 2, 3, 4 or more, and the maximum value of "more" is the maximum number of conditions recorded in each claim. For example, if a claim records 8 conditions, then "one or more" in "satisfying one or more of the following conditions" in that claim is any integer from 1 to 8, such as 1, 2, 3, 4, 5, 6, 7 or 8.

[0089] The term "pharmaceutically acceptable" means that the salts, solvents, excipients, etc., are generally non-toxic, safe, and suitable for patient use. The term "patient" preferably refers to a mammal, and more preferably a human.

[0090] The term "pharmaceutically acceptable salt" refers to a salt prepared from the compounds of this invention with a relatively non-toxic, pharmaceutically acceptable acid or base.

[0091] "Halogen" refers to fluorine, chlorine, bromine, or iodine.

[0092] "Oxo-substituted" means that the oxygen atom replaces two hydrogen atoms on the same carbon atom, that is, the carbonyl group replaces the methylene group.

[0093] "alkyl" (e.g., C 1-14 Alkyl, C 1-8Alkyl, C 1-6 Alkyl groups refer to straight-chain or branched alkyl groups having a specified number of carbon atoms. For example, C1-C2. 14 Alkyl refers to an alkyl group containing 1 to 14 carbon atoms. Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl and similar alkyl groups, preferably methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl or tert-butyl.

[0094] "Alynyl" refers to a straight-chain or branched monovalent alkynyl group having at least two carbon atoms, containing one or more carbon-carbon triple bonds, and without carbon-carbon double bonds. These one or more carbon-carbon triple bonds can be internal or terminal. 2-6 "Alynyl" refers to a straight-chain or branched alkynyl group containing 2-6 carbons, including but not limited to ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, preferably "C". 2-4 "Alynyl", more preferably ethynyl, propynyl or propynyl.

[0095] Unless otherwise specified, the number of atoms in a ring is usually defined as the elemental number of the ring. For example, a “3-10 elemental ring” refers to a “ring” with 3-10 atoms arranged around it.

[0096] "Heterocyclic alkyl" refers to a saturated monocyclic or polycyclic cyclic alkane group having a specified number of ring atoms (e.g., 3-12, 4-12, 3-10, 5-6, 3-8, 3-6, 3-9, 4-6, 5-6, and 5-9), a specified number of heteroatoms (e.g., 1, 2, or 3), and a specified heteroatom type (e.g., one or more of N, O, S, and P, preferably one or more of N, O, and S). "Heterocyclic alkyl" can be a monocyclic or polycyclic heterocyclic alkyl group, wherein the polycyclic heterocyclic alkyl group includes spirocyclic, fused, and bridged cyclic alkyl groups; preferably a monocyclic heterocyclic alkyl group.

[0097] The monocyclic heterocyclic alkyl group is preferably a 3-8 membered monocyclic heterocyclic alkyl group containing one, two or three heteroatoms independently selected from N, O or S; non-limiting examples of monocyclic heterocyclic alkyl groups include azirrobutyl, oxadiol, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.).

[0098] "Alkene heterocycle" refers to a non-aromatic cyclic system having a specified number of ring atoms (e.g., 4-12, 5-6), a specified number of heteroatoms (e.g., 1, 2, or 3), a specified type of heteroatom (one or more of N, O, and S), containing one or more carbon-carbon double bonds and no carbon-carbon triple bonds, preferably a monocyclic system. Monocyclic alkene heterocycles include, but are not limited to, […].

[0099] The term "aryl" refers to an aryl group having a specified number of carbon atoms (e.g., C36). 6-10 Aryl groups are cyclic groups consisting solely of carbon atoms, which can be monocyclic or polycyclic, and each ring is aromatic (following Hückel's rule). Aryl groups include, but are not limited to, phenyl and naphthyl groups.

[0100] "Heteroaryl" refers to a heteroaryl system having a specified number of ring atoms (e.g., 5-12, 5-10, 5-6), a specified number of heteroatoms (e.g., 1, 2, 3, 4, or 5), and a specified heteroatom type (one or more of N, O, and S). "Heteroaryl" can be monocyclic or fused-ring. A fused-ring is a polycyclic group of all carbon atoms in which each ring shares a pair of adjacent carbon atoms with the other rings in the system. When it is a fused-ring, at least one ring is aromatic. The number of rings in a fused-ring can be 2.

[0101] "Heteroaromatic ring" refers to a heteroaromatic cyclic system having a specified number of ring atoms (e.g., 5-12, 5-10, 5-6, 8-10), a specified number of heteroatoms (e.g., 1, 2, 3, 4, or 5), and a specified type of heteroatom (one or more of N, O, and S). It can be a monocyclic or fused ring. A fused ring refers to a polycyclic group consisting of all carbon atoms, where each ring shares a pair of adjacent carbon atoms with other rings in the system. When it is a fused ring, at least one ring is aromatic, and the number of rings in a fused ring can be two. In some embodiments of the present invention, the "heteroaromatic ring" is a pyridine ring.

[0102] "Subunit" refers to a divalent group, such as heterocyclic alkylene referring to a divalent heterocyclic alkylene, arylene referring to a divalent aryl, and heteroarylene referring to a divalent heteroaryl. The heterocyclic alkylene, aryl, and heteroaryl groups are as defined above.

[0103] "Optional" means that the event or environment described below may but does not have to occur, and this description includes situations in which the event or environment may or may not occur. For example, "optionally alkyl-substituted heterocyclic alkyl groups" means that alkyl groups may but do not have to be present, and this description includes cases where heterocyclic alkyl groups are substituted with alkyl groups and cases where heterocyclic alkyl groups are not substituted with alkyl groups.

[0104] The term "pharmaceutical excipients" refers to the excipients and additives used in the production of pharmaceuticals and the preparation of prescriptions. They are all substances contained in pharmaceutical preparations, excluding the active ingredient.

[0105] "Pharmaceutical composition" means a mixture containing one or more of the compounds described herein or their physiologically / pharmacologically acceptable salts or prodrugs, along with other chemical components, such as physiologically / pharmacologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to a living organism, to promote the absorption of the active ingredient, and thus to exert its biological activity.

[0106] The term "prevention" refers to the reduction of the risk of acquiring or developing a disease or disorder.

[0107] The term "treatment" means administering the compound or preparation described in this application to improve or eliminate a disease or one or more symptoms related to said disease, and includes:

[0108] (i) Suppress the disease or disease state, that is, curb its development;

[0109] (ii) Relieve the disease or disease state, even if the disease or disease state subsides.

[0110] The term "therapeutic effective amount" means the amount of the compound of this application used to treat a specific disease, condition, or disorder; (ii) to reduce, improve, or eliminate one or more symptoms of a specific disease, condition, or disorder; or (iii) to prevent or delay the onset of one or more symptoms of a specific disease, condition, or disorder described herein. The amount of the compound of this application constituting a "therapeutic effective amount" varies depending on the compound, the disease state and its severity, the route of administration, and the age of the mammal to be treated, but may routinely be determined by a person skilled in the art based on their own knowledge and the present disclosure.

[0111] The term "patient" refers to any animal, preferably a mammal, that is about to receive or has already received administration of the compound or composition according to embodiments of the invention. The term "mammal" includes any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, pigs, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, and humans, with humans being the most preferred.

[0112] Without violating common sense in the field, the above-mentioned preferred conditions can be combined arbitrarily to obtain various preferred embodiments of the present invention.

[0113] The reagents and raw materials used in this invention are all commercially available.

[0114] The positive and progressive effects of this invention are as follows: the small molecule compound provided by this invention has a good inhibitory effect on PARP1 enzyme, and further, it has a strong inhibitory effect on the proliferation of DLD1 BRCA2- / - cells, while having a very weak inhibitory effect on the proliferation of DLD1 wild-type cells, showing good selectivity. Furthermore, the compound of this application can overcome drug resistance, reduce the dosage, reduce toxic side effects, and achieve the purpose of expanding indications and improving therapeutic effects. Detailed Implementation

[0115] The present invention is further illustrated below by way of embodiments, but the invention is not limited to the scope of the embodiments described herein. Experimental methods in the following embodiments that do not specify specific conditions were performed according to conventional methods and conditions, or as selected according to the product instructions.

[0116] All compounds of this invention can be synthesized by those skilled in the art of organic chemistry using various methods. General synthetic schemes for preparing the compounds of this invention are described below. These schemes are general but do not imply limitation on possible techniques used by those skilled in the art to prepare the compounds disclosed herein. Different methods for preparing the compounds of this invention will be apparent to those skilled in the art. Furthermore, the various steps in the synthesis can be performed alternately and sequentially to obtain one or more desired compounds. The Preparation and Examples sections described below provide examples of preparing the compounds of this invention using the methods described in the general scheme.

[0117] The chemical reactions and synthetic techniques described herein are carried out using the reagents and corresponding solvents described herein, and the corresponding reaction yields will also be affected by the reagents and solvents used. Furthermore, it should be understood that all reaction conditions mentioned in the synthetic methods described below, including the choice of solvent, reaction atmosphere, reaction temperature, experimental duration, and reaction reaction order, should be considered as standard operating conditions for the reaction, which should be readily identifiable by those skilled in the art. This is also understandable to those skilled in the art of organic synthesis. The functional groups present on each part of the molecule must be compatible with the reagents used and the reaction itself. Such limitations where some functional groups on each part of the molecule are incompatible with the reaction conditions must be addressed by alternative methods, which will be obvious to those skilled in the art. It is obvious that it is necessary to determine and adjust the order of synthetic steps, or to select a specific synthetic process scheme, to obtain the compounds required by this invention. This is understandable and readily identifiable to those skilled in the art of organic synthesis. It should also be recognized that another major consideration in designing any synthetic route in this field is the rational selection of protecting groups to protect the tolerance of reactive functional groups present in the compounds described herein. For details, please refer to the book by Greene et al. in the field of chemistry (Protective Groups in Organic Synthesis, Third Edition, Wiley and Sons (1999)).

[0118] Example

[0119] The preparation of compounds and intermediates used in compound preparation can be carried out using the procedures shown in the following examples and related procedures. The methods and conditions used in these examples, and the actual compounds prepared in these examples, are not intended to be limiting, but rather to illustrate how the relevant compounds can be prepared. The starting materials and reagents used in these examples, when not prepared using the procedures described herein, are generally commercially available, or reported in relevant chemical literature, or can be prepared using the procedures described in the chemical literature.

[0120] In the examples given in this article, the term "drying and concentration" typically refers to adding anhydrous sodium sulfate or magnesium sulfate to an organic solvent to dry it, followed by filtration and removal of the solvent from the filtrate (usually under reduced pressure and at a temperature suitable for the stability of the compound being prepared). Column chromatography typically uses conventional or rapid column chromatography for column separation and purification, or a medium-pressure chromatograph (Biotage Isola One) pre-packed with a silica gel column, eluting in a specified solvent or solvent mixture. In some cases, the final product is rapidly purified by preparative thin-layer chromatography using 20 cm x 20 cm x 0.5 mm or 20 cm x 20 cm x 1 mm silica gel plates in a suitable solvent system. Preparative high-performance liquid chromatography (HPLC) is performed using a reversed-phase column (Waters Sunfire C18, Waters Xbridge C18, or similar) with dimensions suitable for the amount of the compound being separated. Elution is typically performed using a gradient of methanol or acetonitrile concentrations in an aqueous phase, with the eluent containing 0.05% or 0.1% formic acid, trifluoroacetic acid, or 10 mM ammonium acetate. The elution rate is matched to the size of the reversed-phase column used and the resolution of the compound being prepared.

[0121] List of abbreviations

[0122] Example 1: 3-Ethyl-7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0123] Step 1: 1-(4-Iodophenyl)piperazine hydrochloride

[0124] At room temperature, 1,4-dioxane hydrochloride (25 mL) was added to a reaction flask containing 1 g (2.58 mmol) of 4-(4-iodophenyl)piperazine-1-carboxylic acid tert-butyl ester. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the mixture was filtered, the filter cake was washed with ethyl acetate, and dried under reduced pressure to give the target compound (white solid, 1 g, yield 96%). LC / MS (ESI) (m / z): 289 [M+H] + .

[0125] Step 2: 3-Ethyl-7-((4-(4-iodophenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0126] At room temperature, 1-(4-iodophenyl)piperazine hydrochloride (1 g, 2.77 mmol), 7-(chloromethyl)-3-ethyl-1,5-naphthidium-2(1H)-one (0.93 g, 4.16 mmol), N,N-diisopropylethylamine (1.79 g, 13.8 mmol), sodium iodide (40 mg, 0.28 mmol), and acetonitrile (30 mL) were added to a reaction flask. The reaction mixture was stirred at 80 °C for 18 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure, and dichloromethane and water were added to the mixture. The layers were separated, and the aqueous phase was extracted with dichloromethane. The combined organic phases were washed successively with water and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was slurried with diethyl ether to give the target compound (yellow solid, 1.3 g, 99% yield). LC / MS (ESI) (m / z): 475 [M+H] + .

[0127] Step 3: 3-Ethyl-7-((4-((trimethylsilyl)ethynyl)phenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0128] Under nitrogen protection, 3-ethyl-7-((4-(4-iodophenyl)piperazin-1-yl)methyl)-1,5-naphthid-2(1H)-one (1.3 g, 2.74 mmol), trimethylethynylsilane (0.54 g, 5.48 mmol), triethylamine (3.5 mL, 25.2 mmol), cuprous iodide (50 mg, 0.274 mmol), palladium dichloride bis(triphenylphosphine) (100 mg, 0.137 mmol), and anhydrous N,N-dimethylformamide (7 mL) were added to a microwave reaction flask. After nitrogen purging, the mixture was microwaved at 100 °C for one hour. The mixture was then concentrated under reduced pressure to remove triethylamine and N,N-dimethylformamide. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-7% methanol) to give the target compound (yellow solid, 1.2 g, 99% yield). LC / MS(ESI) m / z: 445 [M+H] + .

[0129] Step 4: 3-Ethyl-7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0130] Potassium carbonate (1.90 g, 13.7 mmol) was added to a solution of 3-ethyl-7-((4-((trimethylsilyl)ethynyl)phenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one (1.22 g, 2.74 mmol) in anhydrous methanol (20 mL) and tetrahydrofuran (20 mL) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. Methanol and tetrahydrofuran were removed by concentration under reduced pressure. Water and ethyl acetate were added, and the mixture was allowed to separate into layers. The aqueous phase was extracted twice with ethyl acetate. The combined organic phases were washed successively with water and saturated brine, and concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-7% methanol) to give the target compound (yellow solid, 660 mg, yield 64.7%). LC / MS (ESI) m / z: 373 [M+H] + .

[0131] Example 2: 3-Ethyl-7-((4-(4-(hydroxymethyl)phenyl)ethynyl)phenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0132] Step 1: 3-Ethyl-7-((4-(4-(hydroxymethyl)phenyl)ethynyl)phenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0133] 3-Ethyl-7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one (20 mg, 0.054 mmol) was dissolved in a mixed solution of N,N-dimethylformamide (1 mL) and triethylamine (1 mL). (4-iodophenyl)methanol (15 mg, 0.064 mmol), cuprous iodide (1 mg, 0.005 mmol), and bis(triphenylphosphine)palladium dichloride (2 mg, 0.003 mmol) were added to the reaction solution. After purging with nitrogen, the reaction solution was stirred at 100 °C for 3 hours under nitrogen atmosphere. The reaction solution was concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-15% methanol) and then separated by preparative HPLC (C18, 25-95% acetonitrile aqueous solution containing 0.1% NH4HCO3) to obtain the target compound (white solid, 2.43 mg, yield 9.46%). 1H NMR (400MHz, DMSO-d6) δ11.85(s,1H),8.41(d,J=1.5Hz,1H),7.75(s,1H),7.63(s,1H),7.45(d,J=8.1Hz,2H),7.38–7.32(m,4H),6.94(d,J=8.9Hz,2 H),5.29-5.22(m,1H),4.51(d,J=5.7Hz,2H),3.64(s,2H),3.26-3.21(m,4 H),2.55-2.54(m,6H),1.18(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):479[M+H] + .

[0134] Example 3: 3-Ethyl-7-((4-(6-ethynylpyridin-3-yl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0135] Step 1: 4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester

[0136] Under nitrogen protection, 7-(chloromethyl)-3-ethyl-1,5-naphthyl-2(1H)-one (239 mg, 1.07 mmol), piperazine-1-carboxylic acid tert-butyl ester (200 mg, 1.07 mmol), and N,N-diisopropylethylamine (416 mg, 3.22 mmol) were dissolved in anhydrous acetonitrile (2 mL), followed by the addition of sodium iodide (16 mg, 0.11 mmol). The reaction mixture was stirred at 80 °C for 1 hour. After the reaction was complete, a solid precipitated. The reaction mixture was filtered, the filter cake was washed with acetonitrile, and the filter cake was dried under reduced pressure to give the target compound (white solid, 280 mg, yield 70.0%). LC / MS (ESI) m / z: 373 [M+H] + .

[0137] Step 2: 3-Ethyl-7-(piperazin-1-ylmethyl)-1,5-naphthidium-2(1H)-one

[0138] 4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)piperazine-1-carboxylic acid tert-butyl ester (280 mg, 0.75 mmol) was added to a 4M dioxane hydrochloride (3 mL) reaction flask. The reaction mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to give the target compound (white solid, 180 mg, yield 87.9%). LC / MS (ESI) m / z: 273 [M+H] + .

[0139] Step 3: 5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)piperazin-1-yl)pyridinecarboxaldehyde

[0140] Under nitrogen protection, 3-ethyl-7-(piperazin-1-ylmethyl)-1,5-naphthid-2(1H)-one (60 mg, 0.22 mmol) and 5-fluoropyridin-2-aldehyde (28 mg, 0.22 mmol) were dissolved in anhydrous dimethyl sulfoxide (1 mL), followed by the addition of N,N-diisopropylethylamine (85 mg, 0.66 mmol). The reaction mixture was stirred at 80 °C for 2 hours. After the reaction was complete, ethyl acetate and water were added, the ethyl acetate phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-15% methanol) to give the target compound (white solid, 60 mg, yield 72.1%). LC / MS (ESI) m / z: 378 [M+H] + .

[0141] Step 4: 3-Ethyl-7-((4-(6-ethynylpyridin-3-yl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0142] Under nitrogen protection, 5-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthid-3-yl)methyl)piperazin-1-yl)pyridinecarboxaldehyde (60 mg, 0.16 mmol) was dissolved in anhydrous methanol (1 mL), and potassium carbonate (66 mg, 0.48 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (92 mg, 0.48 mmol) were added. The mixture was then stirred at room temperature for 1 hour. After the reaction was complete, the mixture was filtered, and the filtrate was purified by preparative HPLC (20-95% acetonitrile in H2O solution and 0.1% ammonium bicarbonate) to give the target compound (white solid, 14.7 mg, yield 24.8%). 1 H NMR (400MHz, DMSO-d6) δ11.83(s,1H),8.40(d,J=1.7Hz,1H),8.26(d,J=2.8Hz,1H),7.75(s,1H),7.62(s,1H),7.36(d,J=8.7Hz,1H),7.26( dd,J=8.8,3.0Hz,1H),4.05(s,1H),3.64(s,2H),3.29-3.26(m,4H),2.58-2.52(m,6H),1.18(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):374[M+H] + .

[0143] Example 4: 7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0144] Step 1: 1-(4-Iodophenyl)piperazine

[0145] 4-(4-iodophenyl)piperazine-1-carboxylic acid tert-butyl ester (1 g, 2.6 mmol) was dissolved in hydrochloric acid / 1,4-dioxane (10 mL) solution at room temperature, and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction mixture was concentrated to give the target compound (white solid, 720 mg, yield 96.1%). LC / MS (ESI) m / z: 289 [M+H] + .

[0146] Step 2: 8-Fluoro-7-((4-(4-iodophenyl)piperazin-1-yl)methyl)-3-methylquinoxalin-2(1H)-one

[0147] To anhydrous acetonitrile (2 mL) of 1-(4-iodophenyl)piperazine (60 mg, 0.21 mmol) and 7-(bromomethyl)-8-fluoro-3-methylquinoxalin-2(1H)-one (56 g, 0.21 mmol), N,N-diisopropylethylamine (81 mg, 0.63 mmol) was added, and the reaction mixture was stirred at 80 °C for 2 h. After the reaction was complete, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and then filtered and concentrated. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 50-70% ethyl acetate) to give the target compound (yellow solid, 69 mg, yield 69.3%). LC / MS (ESI) m / z: 479 [M+H] + .

[0148] Step 3: 8-Fluoro-3-methyl-7-((4-((trimethylsilyl)ethyl)phenyl)piperazin-1-yl)methyl)quinoxalin-2(1H)-one

[0149] Under nitrogen protection, cuprous iodide (3 mg, 0.014 mmol) and palladium dichloride (10 mg, 0.014 mmol) were added to a solution of 8-fluoro-7-((4-(4-iodophenyl)piperazin-1-yl)methyl)-3-methylquinoxalin-2(1H)-one (69 mg, 0.144 mmol) and ethynyltrimethylsilane (71 mg, 0.722 mmol) in N,N-dimethylformamide (1 mL) and triethylamine (1 mL). The reaction mixture was purged with nitrogen three times, and then stirred at 100 °C for 2 hours under nitrogen protection. After the reaction was complete, the mixture was cooled to room temperature, extracted with water and ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 50-70% ethyl acetate) to give the target compound (yellow solid, 53 mg, yield 82.2%). LC / MS(ESI) m / z: 449(M+H) + .

[0150] Step 4: 7-((4-(4-ethylphenyl)piperazin-1-yl)methyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0151] 8-Fluoro-3-methyl-7-((4-((trimethylsilyl)ethyl)phenyl)piperazin-1-yl)methyl)quinoxalin-2(1H)-one (53 mg, 0.12 mmol) was dissolved in methanol (1 mL) at room temperature. Potassium carbonate (81 mg, 0.59 mmol) was then added to the reaction solution, and the mixture was stirred at room temperature for 2 hours. After the reaction was complete, the reaction solution was filtered and purified by preparative HPLC (C18, 10-50% acetonitrile in H2O solution and 0.1% ammonium bicarbonate) to obtain the target compound (8 mg, white solid, yield 18.0%). 1 H NMR (400MHz, DMSO-d6) δ7.52(d,J=8.3Hz,1H),7.29(d,J=8.5Hz,3H),6.89(d,J=7.4Hz,2H),3.91(s ,1H),3.68(s,2H),3.24-3.17(m,4H),2.56-2.53(m,4H),2.42(s,3H).LC / MS(ESI)(m / z):377[M+H] + .

[0152] Example 5: 7-((4-(6-ethynyl-2-fluoropyridin-3-yl)piperazin-1-yl)methyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0153] Step 1: 4-(2-fluoro-6-(methoxy(methyl)carbamoyl)pyridin-3-yl)piperazine-1-carboxylic acid tert-butyl ester

[0154] Under nitrogen protection, 5-(4-(tert-butyloxycarbonyl)piperazin-1-yl)-6-fluoropyridine acid (100 mg, 0.31 mmol) was dissolved in anhydrous dichloromethane (2 mL), and N,N'-carbonyldiimidazole (55 mg, 0.34 mmol) was added. The mixture was stirred at room temperature for 2 hours, followed by the addition of dimethylhydroxylamine hydrochloride (28 mg, 0.46 mmol) and triethylamine (47 mg, 0.46 mmol), and the reaction was continued at room temperature with stirring for another 16 hours. After the reaction was complete, dichloromethane and water were added, the dichloromethane phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0-80% ethyl acetate) to give the target compound (white solid, 100 mg, yield 88.3%). LC / MS (ESI) m / z: 369 [M+H] + .

[0155] Step 2: 4-(2-fluoro-6-formylpyridin-3-yl)piperazine-1-carboxylic acid tert-butyl ester

[0156] Under nitrogen protection, 100 mg (0.27 mmol) of 4-(2-fluoro-6-(methoxy(methyl)carbamoyl)pyridin-3-yl)piperazine-1-carboxylic acid tert-butyl ester was dissolved in anhydrous tetrahydrofuran (2 mL), and the mixture was cooled to -78 °C and diisobutylaluminum hydride (0.4 mL, 0.41 mmol, 1 M) was slowly added dropwise. After the addition was complete, the mixture was stirred at -78 °C for 1 hour. After the reaction was complete, methanol was added at -78 °C to quench the reaction, followed by the addition of ethyl acetate and water. The ethyl acetate phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-15% methanol) to give the target compound (white solid, 60 mg, yield 71.4%). LC / MS (ESI) m / z: 310 [M+H] + .

[0157] Step 3: 6-Fluoro-5-(piperazin-1-yl)pyridinecarboxaldehyde

[0158] 60 mg (0.19 mmol) of 4-(2-fluoro-6-formylpyridin-3-yl)piperazine-1-carboxylic acid tert-butyl ester was added to a 4 M dioxane hydrochloride (2 mL) reaction flask. The reaction mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure to give the target compound (white solid, 40 mg, yield 98.5%). LC / MS (ESI) m / z: 210 [M+H] + .

[0159] Step 4: 6-Fluoro-5-(4-((5-Fluoro-2-methyl-3-oxo-3,4-dihydroquinoxalo-6-yl)methyl)piperazin-1-yl)pyridinecarboxaldehyde

[0160] Under nitrogen protection, 6-fluoro-5-(piperazin-1-yl)pyridinecarboxaldehyde (39 mg, 0.18 mmol), 7-(bromomethyl)-8-fluoro-3-methylquinoxalin-2(1H)-one (50 mg, 0.18 mmol), and N,N-diisopropylethylamine (95 mg, 0.74 mmol) were dissolved in anhydrous acetonitrile (2 mL), followed by the addition of sodium iodide (3 mg, 0.018 mmol). The reaction mixture was stirred at 80 °C for 2 hours. After the reaction was complete, ethyl acetate and water were added, the ethyl acetate phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-15% methanol) to give the target compound (white solid, 65 mg, yield 88.2%). LC / MS (ESI) m / z: 400 [M+H] + .

[0161] Step 5: 7-((4-(6-ethynyl-2-fluoropyridin-3-yl)piperazin-1-yl)methyl)-8-fluoro-3-methylquinoxalin-2(1H)-one

[0162] Under nitrogen protection, 6-fluoro-5-(4-((5-fluoro-2-methyl-3-oxo-3,4-dihydroquinoxalino-6-yl)methyl)piperazin-1-yl)pyridinecarboxaldehyde (65 mg, 0.16 mmol) was dissolved in anhydrous methanol (1 mL), and potassium carbonate (67 mg, 0.48 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (94 mg, 0.48 mmol) were added. The mixture was then stirred at room temperature for 1 hour. After the reaction was complete, the mixture was filtered, and the filtrate was purified by preparative HPLC (20-95% acetonitrile in H2O solution and 0.1% ammonium bicarbonate) to give the target compound (white solid, 36.4 mg, yield 56.5%). 1 H NMR (400MHz, DMSO-d6) δ12.43(s,1H),7.51(d,J=8.3Hz,1H),7.42(d,J=5.5Hz,2H),7.30-7.25(m,1H),4 .25(s,1H),3.68(s,2H),3.14-3.08(m,4H),2.60-2.55(m,4H),2.41(s,3H).LC / MS(ESI)(m / z):396[M+H] + .

[0163] Example 6: 3-Ethyl-7-((4-(5-ethynylpyrimidin-2-yl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0164] Step 1: 3-Ethyl-7-((4-(5-ethynylpyrimidin-2-yl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one

[0165] Potassium carbonate (51 mg, 0.37 mmol) was added to a solution of 3-ethyl-7-(piperazin-1-ylmethyl)-1,5-naphthyl-2(1H)-one (50 mg, 0.18 mmol) and 2-chloro-5-ethylpyrimidine (51 mg, 0.37 mmol) in N-methylpyrrolidone (1 mL) at room temperature. The reaction mixture was stirred at 80 °C for 16 hours. After the reaction was complete, water was added, and the mixture was extracted with ethyl acetate. The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and then filtered and concentrated. The residue was separated by preparative HPLC (C18, 10-50% acetonitrile in H2O solution and 0.1% ammonium bicarbonate) to obtain the target compound (white solid, 8 mg, yield 11.6%). 1 H NMR (400MHz, DMSO-d6) δ11.85(s,1H),8.47(s,2H),8.40(d,J=1.7Hz,1H),7.75(s,1H),7.63(s,1H),4.27(s,1H),3.8 1-3.76(m,4H),3.63(s,2H),2.58-2.53(m,2H),2.48-2.44(m,4H),1.18(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):375[M+H] + .

[0166] Example 7: 3-Ethyl-7-((4-(5-ethynylpyridin-2-yl)piperazin-1-yl)methyl)-1,5-naphthyridin-2(1H)-one

[0167] Step 1: 3-Ethyl-7-((4-(5-ethynylpyridin-2-yl)piperazin-1-yl)methyl)-1,5-naphthyridin-2(1H)-one

[0168] Under nitrogen protection, 3-ethyl-7-(piperazin-1-ylmethyl)-1,5-naphthidium-2(1H)-one (60 mg, 0.22 mmol) and triethylamine (67 mg, 0.66 mmol) were dissolved in anhydrous dimethyl sulfoxide (1 mL), followed by the addition of 5-ethynyl-2-fluoropyridine (27 mg, 0.22 mmol). The mixture was then stirred at 80 °C for 6 hours. After the reaction was complete, the mixture was filtered, and the filtrate was purified by preparative HPLC (20-95% acetonitrile in H₂O solution and 0.1% ammonium bicarbonate) to obtain the target compound (white solid, 15 mg, yield 18.2%). 1 H NMR (400MHz, DMSO-d6) δ11.63(s,1H),8.40(d,J=1.7Hz,1H),8.21(d,J=2.2Hz,1H),7.75(s,1H),7.62(s,1H),7.58(dd,J=8.9,2.3Hz,1H),6.81(d,J =8.9Hz,1H),4.08(s,1H),3.63(s,2H),3.58-3.53(m,4H),2.57-2.53(m,2 H),2.49-2.45(m,4H),1.18(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):374[M+H] + .

[0169] Example 8: 2”-(difluoromethyl)-N-(6-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthylpyridin-3-yl)methyl)piperazin-1-yl)phenyl)ethynyl)benzo[d]thiazo-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-terpyridine]-5'-formamide

[0170] Step 1: 2”-(difluoromethyl)-3-fluoro-N-(6-iodobenzo[d]thiazo-2-yl)-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-formamide

[0171] At room temperature, 6-iodobenzo[d]thiazol-2-amine (50 mg, 0.181 mmol), 2”-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxylic acid (71 mg, 0.181 mmol), and N-methylimidazole (90 mg, 1.09 mmol) were dissolved in acetonitrile (5 mL). After stirring at 70 °C for 1 min, a solution of N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (76 mg, 0.272 mmol) in acetonitrile (1 mL) was added to the reaction solution. The reaction solution was then stirred at 70 °C for 1 h, and a solid precipitated. The reaction solution was filtered, the filter cake was washed with acetonitrile, and dried under reduced pressure to give the target compound (yellow solid, 80 mg, yield 68%). LC / MS (ESI) m / z: 649.8 [M+H] + .

[0172] Step 2: 2”-(difluoromethyl)-N-(6-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthylpyridin-3-yl)methyl)piperazin-1-yl)phenyl)ethynyl)benzo[d]thiazo-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-terpyridine]-5'-formamide

[0173] Under nitrogen protection at room temperature, 2”-(difluoromethyl)-3-fluoro-N-(6-iodobenzo[d]thiazo-2-yl)-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxamide (80 mg, 0.123 mmol), 3-ethyl-7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one (55.06 mg, 0.148 mmol), triethylamine (3 mL), cuprous iodide (2.35 mg, 0.012 mmol), and ditriphenylphosphine dichlorophosphate were added. Palladium (4.32 mg, 0.006 mmol) and N,N-dimethylformamide (3 mL) were added to a reaction flask, purged three times with nitrogen, and stirred at 100 °C for 3 hours. The mixture was concentrated under reduced pressure to remove triethylamine and N,N-dimethylformamide. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-5% methanol) to obtain the crude compound. The crude product was slurried with methanol and then purified by preparative HPLC (eluent: acetonitrile / 0.1% ammonium bicarbonate aqueous solution, gradient: 25%–90%) to obtain the target compound (yellow solid, 25 mg, yield 22.7%). 1H NMR (400MHz, DMSO-d6) δ13.30(s,1H),11.86(s,1H),9.05(s,1H),8.52(s,1H),8.42(d,J=1.7Hz, 1H),8.17(s,1H),8.10(s,1H),7.88(d,J=7.0Hz,1H),7.80-7.76(m,3H),7.64(s,1H),7.60-7.55 (m,2H),7.40(d,J=8.8Hz,2H),7.16-6.88(m,3H),6.44(td,J=7.3,4.7Hz,1H),3.73(s,3H),3.66 (s,2H),3.27-3.21(m,4H),2.58-2.54(m,6H),1.19(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):894[M+H] + .

[0174] Example 9: 2'-(difluoromethyl)-N-(6-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthylpyridin-3-yl)methyl)piperazin-1-yl)phenyl)ethynyl)benzo[d]thiazolyl-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

[0175] Step 1: 2'-(difluoromethyl)-N-(6-iodobenzo[d]thiazolyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

[0176] Under nitrogen protection, 2'-(difluoromethyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxylic acid (30 mg, 0.10 mmol), 6-iodobenzo[d]thiazol-2-amine (28 mg, 0.10 mmol), and N-methylimidazole (42 mg, 0.51 mmol) were dissolved in anhydrous acetonitrile (2 mL). After stirring at 70 °C for 5 minutes, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (45 mg, 0.20 mmol) was added. The reaction mixture was stirred at 70 °C for 1 hour. After the reaction was complete, a white solid product precipitated, which was filtered, and the filter cake was washed with a small amount of acetonitrile and dried to give the target compound (white solid, 33 mg, yield 58.6%). LC / MS (ESI) m / z: 553 [M+H] + .

[0177] Step 2: 2'-(difluoromethyl)-N-(6-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)piperazin-1-yl)phenyl)ethynyl)benzo[d]thiazolyl-2-yl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide

[0178] Under nitrogen protection, 3-ethyl-7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-1,5-naphthidin-2(1H)-one (22 mg, 0.06 mmol) and 2'-(difluoromethyl)-N-(6-iodobenzo[d]thiazolyl)-5'-methoxy-6-methyl-[4,4'-bipyridine]-3-carboxamide (33 mg, 0.06 mmol) were dissolved in a sealed tube containing anhydrous N,N-dimethylformamide (0.5 mL). Triethylamine (0.5 mL), cuprous iodide (2 mg, 0.012 mmol), and palladium dichloride bis(triphenylphosphine) (4 mg, 0.006 mmol) were added. The reaction mixture was purged three times under nitrogen purging and stirred at 100 °C for 1 hour. After the reaction was complete, N,N-dimethylformamide was dried, and the residue was subjected to silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-15% methanol) to obtain crude product. The crude product was purified by preparative HPLC (20-95% acetonitrile in H2O solution and 0.1% ammonium bicarbonate) to obtain the target compound (white solid, 11.7 mg, yield 24.7%). 1 H NMR (400MHz, DMSO-d6) δ13.09(s,1H),11.85(s,1H),8.87(s,1H),8.47(s,1H),8.41(d,J=1 .7Hz,1H),8.14(s,1H),7.76(t,J=7.6Hz,3H),7.64(s,1H),7.56(dd,J=8.4,1.5Hz,1H),7. 47(s,1H),7.40(d,J=8.8Hz,2H),7.14-6.86(m,3H),3.69(s,3H),3.65(s,2H),3.27-3.23( m,4H),2.62(s,3H),2.58-2.53(m,6H),1.19(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):797[M+H] + .

[0179] Example 10: 2”-(difluoromethyl)-N-(5-((4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthidin-3-yl)methyl)piperazin-1-yl)phenyl)ethynyl)-1,3,4-thiadiazol-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-terpyridine]-5'-formamide

[0180] Step 1: N-(5-bromo-1,3,4-thiadiazol-2-yl)-2”-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-terpyridine]-5'-carboxamide

[0181] At room temperature, 1-propyl phosphate cyclic anhydride (407 mg, 0.64 mmol, 50% Wt ethyl acetate) was added to an anhydrous pyridine solution of 2”-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-terpyridine]-5'-carboxylic acid (50 mg, 0.13 mmol) and 5-bromo-1,3,4-thiadiazol-2-amine (23 mg, 0.13 mmol). The reaction mixture was stirred at 80 °C for 3 h. The mixture was concentrated to dryness under reduced pressure. The residue was purified by silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-5% methanol) to give the target compound (white solid, 50 mg, yield 70.7%). LC / MS (ESI) m / z: 553 [M+H] + .

[0182] Step 2: 2”-(difluoromethyl)-N-(5-((4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthidin-3-yl)methyl)piperazin-1-yl)phenyl)ethynyl)-1,3,4-thiadiazol-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-terpyridine]-5'-formamide

[0183] Under nitrogen protection, N-(5-bromo-1,3,4-thiadiazol-2-yl)-2”-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-terpyridine]-5'-carboxamide (25 mg, 0.045 mmol) was dissolved in a mixed solution of 1,4-dioxane (5 mL) and water (1.5 mL). 3-ethyl-7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-1,5-naphthidium-2(1H)-one (20 mg, 0.054 mmol) was added to the reaction solution. Lithium carbonate (10 mg, 0.135 mmol), cuprous iodide (1 mg, 0.005 mmol), and tetraphenylphosphine palladium (5 mg, 0.004 mmol) were used. The reaction solution was purged three times under nitrogen and then stirred at 100 °C for 3 hours. After the reaction was complete, ethyl acetate and water were added, the ethyl acetate phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-10% methanol) followed by preparative HPLC (C18, 25-95% acetonitrile in aqueous solution containing 0.1% NH4HCO3) to obtain the target compound (yellow solid, 8.43 mg, yield 22.1%). 1 H NMR(400MHz,DMSO-d6)δ13.65(s,1H),11.88(s,1H),9.04(s,1H),8.51(s,1H),8.42(s,1H),8.0 9(s,1H),7.87(d,J=7.1Hz,1H),7.75(s,2H),7.65(s,1H),7.60-7.55(m,1H),7.49(d,J=8.7Hz, 2H),7.01(t,J=55.0Hz,1H),7.00(d,J=8.2Hz,2H),6.46-6.40(m,1H),3.70(s,3H),3.50-3.34( m,2H),3.30-3.24(m,4H),2.59-2.53(m,6H),1.19(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):845[M+H] + .

[0184] Example 11: 2”-(difluoromethyl)-N-(5-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthylpyridin-3-yl)methyl)piperazin-1-yl)phenyl)

[0185] (ethynyl)thiazolyl-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxamide

[0186] Step 1: N-(5-bromothiazol-2-yl)-2'-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxamide

[0187] At room temperature, 5-bromothiazol-2-amine (30 mg, 0.17 mmol), 2”-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxylic acid (66 mg, 0.17 mmol), and N-methylimidazole (82.55 mg, 1.01 mmol) were dissolved in acetonitrile (5 mL). After stirring at 70 °C for 1 minute, N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (7 mL) was added to the reaction solution. A solution of 0.53 mg (0.25 mmol) of acetonitrile (1 mL) was prepared, and the reaction mixture was stirred at 70 °C for one hour. The acetonitrile was removed by concentration under reduced pressure. The residue was dissolved in dichloromethane, washed with water and brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was then subjected to silica gel column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0-50% ethyl acetate) to give the target compound (white solid, 60 mg, yield 64.8%). LC / MS (ESI) m / z: 552 [M+H] + .

[0188] Step 2: 2”-(difluoromethyl)-N-(5-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthyridin-3-yl)methyl)piperazin-1-yl)phenyl)ethynyl)thiazo-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-formamide

[0189] Under nitrogen protection, N-(5-bromothiazol-2-yl)-2'-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxamide (40 mg, 0.072 mmol), 3-ethyl-7-((4-(4-ethynylphenyl)piperazin-1-yl)methyl)-1,5-naphthoidin-2(1H)-one (32.37 mg, 0.087 mmol), cuprous iodide (6.90 mg, 0.036 mmol), lithium carbonate (10.70 mg, 0.145 mmol), tetrakis(triphenylphosphine)palladium (12.55 mg, 0.011 mmol), 1,4-dioxane (9 mL), and water (3 mL) were added to a reaction flask. After purging with nitrogen, the mixture was stirred at 100 °C for 3 hours. The residue was concentrated under reduced pressure and purified by silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0-5% methanol) to obtain the crude compound, which was then purified by preparative HPLC (eluent: acetonitrile / 0.1% ammonium bicarbonate aqueous solution, gradient: 25%-90%) to obtain the target compound (yellow solid, 13 mg, yield 21.3%). 1 H NMR (400MHz, DMSO-d6) δ13.24(s,1H),11.85(s,1H),9.00(s,1H),8.52(s,1H),8.41(d,J=1.7Hz,1 H),8.08(s,1H),7.87(d,J=7.1Hz,1H),7.79(s,1H),7.75(d,J=3.8Hz,2H),7.63(s,1H),7.61-7.5 4(m,1H),7.36(d,J=8.8Hz,2H),7.15-6.87(m,3H),6.43(td,J=7.3,4.7Hz,1H),3.71(s,3H),3.65 (s,2H),3.28-3.24(m,4H),2.58-2.54(m,6H),1.19(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):844[M+H] + .

[0190] Example 12: 2”-(difluoromethyl)-N-(6-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthylpyridin-3-yl)methyl)piperazin-1-yl)-3-fluorophenyl)ethynyl)benzo[d]thiazo-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-tripyridine]-5'-formamide

[0191] Step 1: 2”-(difluoromethyl)-3-fluoro-N-(6-iodobenzo[d]thiazo-2-yl)-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-formamide

[0192] Under nitrogen protection, 2”-(difluoromethyl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxylic acid (200 mg, 0.51 mmol), 6-iodobenzo[d]thiazol-2-amine (70 mg, 0.51 mmol), and N-methylimidazole (209 mg, 2.55 mmol) were dissolved in anhydrous acetonitrile (2 mL). The mixture was stirred at 70 °C for 5 minutes, and then N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (286 mg, 5.10 mmol) was added. The reaction mixture was stirred at 70 °C for 1 hour. After the reaction was complete, a solid precipitated out. The solid was filtered, washed with acetonitrile, and dried to give the target compound (white solid, 220 mg, yield 66.4%). LC / MS (ESI) m / z: 650 [M+H] + .

[0193] Step 2: 4-(2-fluoro-4-formylphenyl)piperazine-1-carboxylic acid tert-butyl ester

[0194] 3,4-Difluorobenzaldehyde (2 g, 14.07 mmol) and piperazine-1-carboxylic acid tert-butyl ester (2.6 g, 14.01 mmol) were dissolved in anhydrous dimethyl sulfoxide (20 mL), followed by the addition of N,N-diisopropylethylamine (3.6 g, 28.15 mmol). The reaction mixture was stirred at 80 °C for 1 hour. After the reaction was complete, ethyl acetate and water were added, the ethyl acetate phase was separated, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0-50% ethyl acetate) to give the target compound (white solid, 3.0 g, yield 69.1%). LC / MS (ESI) m / z: 309 [M+H] + .

[0195] Step 3: 4-(4-ethynyl-2-fluorophenyl)piperazine-1-carboxylic acid tert-butyl ester

[0196] Under nitrogen protection, 200 mg (0.53 mmol) of 4-(2-fluoro-4-formylphenyl)piperazine-1-carboxylic acid tert-butyl ester was dissolved in anhydrous methanol (2 mL), followed by the addition of potassium carbonate (269 mg, 1.95 mmol) and dimethyl (1-diazo-2-oxopropyl)phosphonate (374 mg, 1.95 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in dichloromethane, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was subjected to silica gel column chromatography (eluent: petroleum ether / ethyl acetate, gradient: 0-45% ethyl acetate) to give the target compound (white solid, 190 mg, yield 96.2%). LC / MS (ESI) m / z: 305 [M+H] + .

[0197] Step 4: 4-(4-((2-(2”-(difluoromethyl)-3-fluoro-5'-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxamide)benzo[d]thiazolyl-6-yl)ethynyl)-2-fluorophenyl)piperazine-1-carboxylic acid tert-butyl ester

[0198] Under nitrogen protection, 2”-(difluoromethyl)-3-fluoro-N-(6-iodobenzo[d]thiazo-2-yl)-5”-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxamide (80 mg, 0.12 mmol) and 4-(4-ethynyl-2-fluorophenyl)piperazine-1-carboxylic acid tert-butyl ester (56 mg, 0.19 mmol) were dissolved in a sealed tube containing anhydrous N,N-dimethylformamide (0.5 mL). Triethylamine (0.5 mL), cuprous iodide (5 mg, 0.025 mmol), and palladium dichloride bis(triphenylphosphine) (8 mg, 0.012 mmol) were added. The reaction mixture was purged with nitrogen three times and stirred at 100 °C for 1 hour. After the reaction was complete, N,N-dimethylformamide was dried, and the residue was subjected to silica gel column chromatography (eluent: dichloromethane / methanol, gradient: 0). The target compound was obtained (white solid, 60 mg, yield 58.9%) from 15% methanol. LC / MS (ESI) m / z: 826 [M+H] + .

[0199] Step 5: 2”-(difluoromethyl)-3-fluoro-N-(6-((3-fluoro-4-(piperazin-1-yl)phenyl)ethynyl)benzo[d]thiazo-2-yl)-5”-methoxy-2-oxo-2H-[1,2':4',4”-tripyridine]-5'-formamide

[0200] Under nitrogen protection, 60 mg (0.073 mmol) of 4-(4-((2-(2”-(difluoromethyl)-3-fluoro-5'-methoxy-2-oxo-2H-[1,2':4',4”-bipyridine]-5'-carboxamide)benzo[d]thiazolyl-6-yl)ethynyl)-2-fluorophenyl)piperazine-1-carboxylic acid tert-butyl ester was added to a reaction flask containing 2 mL of anhydrous dichloromethane, followed by the addition of trimethylsilyl trifluoromethanesulfonate (32 mg, 0.15 mmol) in an ice bath. The reaction mixture was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was subjected to reverse-phase column chromatography (20-95% methanol in H2O solution and 0.1% FA) to give the target compound (white solid, 44 mg, yield 84.6%). LC / MS (ESI) m / z: 726 [M+H] + .

[0201] Step 6: 2”-(difluoromethyl)-N-(6-(4-(4-((7-ethyl-6-oxo-5,6-dihydro-1,5-naphthylpyridin-3-yl)methyl)piperazin-1-yl)-3-fluorophenyl)ethynyl)benzo[d]thiazo-2-yl)-3-fluoro-5”-methoxy-2-oxo-2H-[1,2':4',4”-tripyridine]-5'-formamide

[0202] Under nitrogen protection, 2”-(difluoromethyl)-3-fluoro-N-(6-((3-fluoro-4-(piperazin-1-yl)phenyl)ethynyl)benzo[d]thiazo-2-yl)-5”-methoxy-2-oxo-2H-[1,2':4',4”-tripyridine]-5'-carboxamide (22 mg, 0.028 mmol) and 7-(chloromethyl)-3-ethyl-1,5-naphthidium-2(1H)-one (7 mg, 0.030 mmol) were mixed. The target compound (0.45 mg, 0.003 mmol) was dissolved in anhydrous acetonitrile (1 mL), and sodium iodide (0.45 mg, 0.003 mmol) and N,N-diisopropylethylamine (12 mg, 0.090 mmol) were added. The reaction mixture was stirred at 80 °C for 2 hours. After the reaction was complete, the mixture was filtered, and the filtrate was purified by preparative HPLC (20-95% acetonitrile in H2O solution and 0.1% ammonium bicarbonate) to give the target compound (white solid, 6 mg, yield 21.7%). 1H NMR(400MHz,DMSO-d6)δ13.32(s,1H),11.86(s,1H),9.05(s,1H),8.52(s,1H),8.41(s,1H) ,8.19(s,1H),8.09(s,1H),7.88(d,J=7.2Hz,1H),7.82-7.73(m,3H),7.63(s,1H),7.62-7.5 4(m,2H),7.37-7.29(m,2H),7.16-6.87(m,2H),6.48-6.40(m,1H),3.72(s,3H),3.66(s,2H ),3.15-3.07(m,4H),2.62-2.53(m,6H),1.19(t,J=7.4Hz,3H).LC / MS(ESI)(m / z):912[M+H] + .

[0203] Effect Example

[0204] Experimental methods:

[0205] 1. Polθ biochemical activity assay (ADP Glo assay)

[0206] The N-terminus of the Polθ protein contains a helicase domain with ATPase activity, capable of hydrolyzing ATP into ADP, which can be detected by the ADP-Glo ​​assay kit (ADP-Glo). TM The Kinase Assay (Promega, V9102) was used to analyze the ATPase activity of Polθ protein and the inhibitory effect of small molecule compounds on the protein. The specific detection method is as follows:

[0207] (1) The helicase domain of the Polθ protein was purified using an insect system to obtain a protein with a purity >90%. ssDNA (5'-CCAGTGAATTGTTGCTCGGTACCTGCTAAC-3', Hangzhou Youkang Biotechnology Co., Ltd.) was ordered as the substrate for the Polθ protein; a reaction buffer containing 10 mM DTT (dithiothreitol), 20 mM MgCl2, Tris-HCl, and pH 7.5 was prepared.

[0208] (2) Prepare 2x ssDNA-Polθ premix and perform the reaction in a 384-well plate. Set up a control group with only buffer and add the premix to the other groups. Add the compound using an automated pipette (Thermo, Multidrop 8) at a starting concentration of 0.5 μM and dilute by 1 / 3. Set up a total of 9 detection points, with 2 replicates per group;

[0209] (3) Prepare 2xATP solution, add an equal volume of 2xATP solution to all wells, and let stand at room temperature for 60 min;

[0210] (4) According to the Promega reagent instructions, add ADP-Glo ​​Detection Reagent to the reaction system and let it stand at room temperature for 60 min;

[0211] (5) According to the Promega reagent instructions, add Kinase Detection Reagent to the reaction system, let it stand at room temperature for 60 min, and use an ELISA reader (Thermo, Varioskan LUX) to detect the chemiluminescence signal. The reading time interval for each well is set to 1000 ms.

[0212] Data analysis was performed on the measurement results: the CV% of the control group test results should be less than 10%, and the z' value should be greater than 0.5. Data meeting the above quality control results were used to calculate the inhibition rate of the compound (inhibition rate (%) = 100 * (control group average - experimental group) / (control group average - buffer group average). A nonlinear regression was used to fit the inhibition rate curve of the compound and the IC50 value was obtained.

[0213] 2. PARP1 biochemical activity assay

[0214] The inhibitory effect of the compounds on PARP1 enzyme activity was detected using the PARP1 Chemiluminescent Assay Kit (BPS Bioscience, USA, Cat#80569). The experiment was conducted according to the manufacturer's instructions, with adjustments made based on laboratory conditions. The specific method is as follows:

[0215] (1) Prepare a 1× histone mixture in PBS, add 25 μL of histone solution to the ELISA plate and incubate overnight at 4°C (prepare one day before the experiment; to ensure the reliability of the experiment, do not coat in advance);

[0216] (2) Add 100 μL of 1×PBST buffer (0.05% Tween-20 added to 1×PBS) to each well for washing. Wash 3 times and place the experimental plate on clean filter paper to remove the liquid.

[0217] (3) Add 100 μL of blocking solution to each well for sealing, place the experimental plate at room temperature for 90 min, and wash the plate according to step 2.

[0218] (4) Dilute 1×PARP buffer to prepare a mixture of 1×PARP substrate buffer and 1×active DNA. Add the mixture to the experimental plate at a rate of 12.5 μL / well.

[0219] (5) Dilute the compound in a gradient of 1 μM at the initial concentration and 1 / 3 dilution, for a total of 9 concentration points. Add 2.5 μL of the test compound to the experimental plate.

[0220] (6) Prepare PARP1 enzyme solution with 1×PARP buffer, add 10 μL to each well, with a final concentration of 2 nM, and start the enzymatic reaction.

[0221] (7) Shake the experimental plate for 30 seconds, incubate at room temperature for 60 minutes, and then wash the plate according to step 2.

[0222] (8) Dilute biotin-HRP with blocking buffer at a ratio of 1:50, add 25 μL / well to the experimental plate, incubate at room temperature for 30 min, and then wash the plate according to step 2.

[0223] (9) Mix HRP chemiluminescent substrate A and chemiluminescent substrate B on ice at a ratio of 1:1, and add 50 μL of the mixture per well to the experimental plate;

[0224] (10) Read the chemiluminescence readings on the microplate reader (Thermo, Varioskan LUX);

[0225] Data analysis was performed on the measurement results: the CV% of the control group should be less than 20%, and the z' value should be greater than 0.5. Data meeting these quality control requirements were used to calculate the inhibition rate (inhibition rate (%) = 100 * (control group average - experimental group) / (control group average - blank group). A nonlinear regression was used to fit the inhibition rate curve of the compound and obtain the IC50 value.

[0226] 3. PARP2 Biochemical Experiment (PARP2-Tracer FP Assay)

[0227] 1) Add 5 μL of diluted GST-PARP2 (final concentration: 10 nM) to each well of a 384-well reaction plate (Corning, Catalog #: 4514);

[0228] 2) Centrifuge horizontally at 1000 RPM for 1 minute in a centrifuge (Xiangyi, Catalog#: ZL-F-7);

[0229] 3) The compounds dissolved in DMSO were added to 384-well plates using a micro-liquid pipette (Thermo Multidrop Pico8) (final compound concentrations: 30000 nM, 7500 nM, 1875 nM, 469 nM, 117 nM, 29 nM, 7.3 nM, 1.8 nM, 0.46 nM, 0.11 nM);

[0230] 4) Add 5 μL of diluted Tracer reactant (final concentration: 5 nM) to each well of the 384-well reaction plate;

[0231] 5) Incubate at room temperature for 1 hour;

[0232] 6) Place the 384-well plate in a microplate reader (Thermo, Catalog#: VarioskanLUX) for fluorescence signal detection;

[0233] 7) Calculate the average value of the DMSO-treated wells in the plate and use it as the High Control (HC). Calculate the average value of the reaction wells without enzyme in the plate and use it as the Low Control (LC). Inhibition rate (%) = (Signal value Ave_HC - Signal value cmpd) / (Signal value Ave_HC - Signal value Ave - LC) × 100;

[0234] 8) Use GraphPad Prism software to perform four-parameter fitting analysis on the data and determine IC50.

[0235] 4. Cell viability assay 1

[0236] The in vitro efficacy of the inhibitor was assessed using a cell viability assay, specifically the commonly used CTG assay, with CellTiter Glo reagent (Promega, G7573). The specific assay method is as follows:

[0237] 1) Culture DLD1 wild-type (ATCC, CCL-221) and DLD1 BRCA2- / - cells. One day before the assay, digest the cells with trypsin (0.025% Trypsin-EDTA, Hyclone), centrifuge at 1000 rpm for 3 min, and collect the cells. Count the cells using a cell counter (Shanghai Mengwei Biomedical Technology Co., Ltd., SmartCell600A.SC1006), and seed them into 96-well white culture plates at an appropriate seeding ratio.

[0238] 2) 24 hours after cell seeding, the drug was added, which was recorded as Day 0. The compound was added using an automated pipette (Thermo, Multidrop 8). The plate was set as a 96-well plate with an initial concentration of 1 μM and a 1 / 3 dilution ratio. A total of 9 detection points were set, with 2 replicates per group. The cells were incubated at 37°C in a 5% CO2 incubator.

[0239] 3) On Day 7, remove the cell culture plate and add an equal volume of CTG reagent (the CTG reagent needs to be brought back to temperature and premixed according to the reagent instructions). Gently mix for 10 minutes (speed 300) on a constant temperature mixer (Hangzhou Aosheng Instrument Co., Ltd., MSC-100). Perform chemiluminescence detection using an ELISA reader (Thermo, Varioskan LUX).

[0240] 4) Data analysis of the measurement results: The CV% of the control group should be less than 20%, and the z' value should be greater than 0.5. Data meeting the above quality control results were used to calculate cell viability (inhibition rate (%) = 100 * (control group average - experimental group) / (control group average - culture medium control group average). GraphPad Prism8 was used to perform nonlinear regression fitting of the compound's inhibition rate curve and derive the IC50. 50 value.

[0241] Tests showed that the compounds in the examples of this application had a strong inhibitory effect on the proliferation of DLD1 BRCA2- / - cells, but a very weak inhibitory effect on the proliferation of DLD1 wild-type cells, indicating that the compounds in the examples of this application have good selectivity in inhibiting the proliferation of the two cell types.

[0242] Cell Viability Assay 2

[0243] (1) PEO1 cells (ATCC) were cultured in DMEM medium (Gibco, Catalog#: C11995500BT) containing 10% serum (Gibco, Catalog#: 10091148) and 1% penicillin and streptomycin (MACKLIN, Catalog#: Q6532).

[0244] (2) After digesting and resuspending PEO1 cells and counting them, 200 cells per well were seeded in 384-well cell culture plates (Univita, Catalog#: 6007680) and incubated in a CO2 (5%) incubator at 37°C for 24 hours to allow the cells to adhere completely.

[0245] (3) The compounds dissolved in DMSO were added to the cell-containing culture plates using a micro-liquid pipette (Thermo Multidrop Pico8) (final compound concentrations: 30000 nM, 10000 nM, 3000 nM, 1000 nM, 300 nM, 100 nM, 30 nM, 10 nM, 3 nM, 0 nM), and incubated in a CO2 (5%) incubator at 37°C for 7 days.

[0246] (4) Place the culture plate containing cells at room temperature for 30 minutes to allow the plate and its contents to equilibrate to room temperature;

[0247] (5) Add CellTiter-Glo reagent (Promega, Catalog#: G7573) equal to the volume of cell culture medium to each well, mix the contents on a fixed-track shaker for 2 minutes to induce cell lysis;

[0248] (6) After incubating at room temperature for 10 minutes to stabilize the signal value, the microplate reader (Thermo, Catalog#: VarioskanLUX) was used to detect and record the luminescence value;

[0249] (7) Calculate the average value of the wells treated with DMSO in the plate and use it as the High Control (HC). Calculate the average value of the wells containing only cell culture medium in the plate and use it as the Low Control (LC). Inhibition rate (%) = (signal value) Ave_HC -Signal value cmpd ) / (signal value) Ave_HC -Signal value Ave-LC )×100;

[0250] (8) Use GraphPad Prism software to perform four-parameter fitting analysis on the data and determine IC50.

[0251] Data list:

[0252] NT = Not detected;

[0253] 5. Mouse pharmacokinetics experiment

[0254] 1) Experimental objective:

[0255] In this study, ICR female mice were used as test animals. The plasma drug concentrations of the test compound were quantitatively determined at different time points after intravenous or oral administration using LC / MS / MS to evaluate the pharmacokinetic characteristics of the test drug in mice.

[0256] 2) Experimental materials:

[0257] ICR mice (female, 18-35g, 5-10 weeks old, Shanghai Slack).

[0258] 3) Experimental procedures:

[0259] The clarified solution of the test compound (solvent: 8% DMSO + 40% PEG400 (manufacturer: MCE; catalog number: HY-Y0873A; batch number: 159569) + 20% (10% TPGS (manufacturer: MACKLIN; catalog number: V819469-250g; batch number: C15030441)) aqueous solution, concentration: 30 mg / mL) was injected into ICR mice via tail vein (after overnight fasting or feeding) or administered to ICR mice via gavage (after overnight fasting or feeding).

[0260] 30 μL of blood was collected via submandibular vein puncture / canthal puncture at 0 h (before administration) and at 0.083, 0.25, 0.5, 1, 2, 4, 7, and 24 h after administration. The blood was placed in an anticoagulant tube containing heparin sodium, and the mixture was thoroughly vortexed at 4 °C and centrifuged at 6500 rpm for 5 minutes.

[0261] Oral administration via gavage was performed at 0 h (before administration) and at 0.25, 1, 2, 4, 7, and 24 h after administration. Blood was collected from the submandibular vein / canthal puncture and placed in an anticoagulant tube containing heparin sodium. The mixture was thoroughly vortexed and centrifuged at 6500 rpm for 5 minutes.

[0262] Blood drug concentrations were determined using LC-MS / MS, and relevant pharmacokinetic parameters were calculated using the non-compartmental linear logarithmic trapezoidal method. Mean, standard deviation, and coefficient of variation were calculated.

[0263] Experimental results: IV: Injection; PO: Oral administration via gavage.

Claims

1. A compound as shown in Formula I or a pharmaceutically acceptable salt thereof, L 1 For not replaced or by one or more R a Replacement C 2-14 alkynyl group; Each R a Independently for not replaced or by one or more R a1 Substituted 6-10 aryl groups; Each R a1 Independently, C is either unsubstituted or substituted with one or more hydroxyl groups. 1-6 alkyl; L 2 For not replaced or by one or more R P2 Substituted 5-10 arylene groups or unsubstituted or substituted with one or more R groups P2 Substituted 5-10 heteroaryl groups; L 3 For not replaced or by one or more R P2 Substituted 3-12 membered heterocyclic alkyl groups; L 4 For not replaced or by one or more R P2 Replacement C 1-14 Alkylene; Each R P2 Halogens are independent of each other; Ring E is unsubstituted or replaced by one or more R e-1 Substituted 6-10 aryl rings or unsubstituted or substituted with one or more R e-1 Replaced 5-12 heterocyclic aromatic rings; Ring F is unsubstituted or replaced by one or more R e-2 Substituted 4-12 membered olefinic heterocycles; Each R e-1 Independent of halogen or unsubstituted C 1-6 alkyl; Each R e-2 Independent oxo group (=O), halogen or unsubstituted C 1-6 alkyl; The heteroatoms in the 5-10-membered heteroaryl, 3-12-membered heterocyclic alkyl, 5-12-membered heteroaromatic and 4-12-membered olefinic heterocycles are independently selected from one, two or three of N, O and S, and the number of heteroatoms is independently 1, 2, 3, 4 or 5; the number of olefin bonds in the 4-12-membered olefinic heterocycle is 1 or 2.

2. The compound of formula I as claimed in claim 1, characterized in that, It meets one or more of the following conditions: (1)L 1 In, the unsubstituted or unreplaced R a Replacement C 2-14 C in alkynyl group 2-14 The alkynyl group is independently C 2-6 alkynyl group, the C 2-6 The alkynyl group can be a straight-chain alkynyl group, for example... For example (2)R a In, the unsubstituted or unreplaced R a1 The 6-10 aryl group in the substituted 6-10 aryl group is phenyl or naphthyl, for example, phenyl; (3)R a1 In, the unsubstituted or unreplaced R a1-1 Replacement C 1-6 C in alkyl 1-6 The alkyl group is methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, or tert-butyl, for example, methyl; (4)L 2 In, the unsubstituted or unreplaced R P2 The 5-10 arylene group in the substituted 5-10 arylene group is independently phenylene or naphthylene, for example, phenylene, for example (5)L 2 In, the unsubstituted or unreplaced R P2 The 5-10-membered heteroaryl group in the substituted 5-10-membered heteroaryl group is independently a 5-6-membered heteroaryl group; (6)L 2 In, the unsubstituted or unreplaced R P2 The heteroatom in the substituted 5-10-membered heteroaryl group is N, and the number of heteroatoms can be independently 1, 2 or 3; (7)L 3 In, the unsubstituted or unreplaced R P2 The 3-12-membered heterocyclic alkyl group in the substituted 3-12-membered heterocyclic alkyl group is independently a monocyclic ring; (8)L 3 In, the unsubstituted or unreplaced R P2 The 3-12-membered heterocyclic alkyl group in the substituted 3-12-membered heterocyclic alkyl group is independently a 3-8-membered heterocyclic alkyl group, such as a 3-6-membered heterocyclic alkyl group; (9)L 3 In, the unsubstituted or unreplaced R P2 The heteroatoms of the substituted 3-12-membered heterocyclic alkyl group are independently selected from one or both of N and O, and the number of heteroatoms can be independently 1 or 2, for example, the heteroatom is N, and the number of heteroatoms is 1 or 2; (10)L 4 In, the unsubstituted or unreplaced R P2 Replacement C 1-14 C in alkylene 1-14 Alkylene is independently C 1-8 Alkylene, the C 1-8 Alkylenes can be straight-chain alkylenes, for example For example (11) In ring E, the unsubstituted or unreplaced R e-1 The 6-10 membered aromatic ring of the substituted 6-10 membered aromatic ring is a benzene ring or a naphthalene ring, for example, a benzene ring; (12) In ring E, the unsubstituted or unreplaced R e-1 The substituted 5-12-membered heteroaryl ring is a 5-10-membered heteroaryl ring, such as a 5-6-membered heteroaryl ring, such as a pyridine ring; (13) In ring E, the heteroatom of each of the 5-12 membered heteroaromatic rings is N, and the number of heteroatoms can be 1, 2 or 3; (14) In ring F, the unsubstituted or unreplaced R e-2 The 4-12-membered olefinic heterocycle in the substituted 4-12-membered olefinic heterocycle is independently a 5-6-membered olefinic heterocycle; (15) In ring F, the unsubstituted or unreplaced R e-2 The heteroatom in the substituted 4-12-membered alkene heterocycle is N, and the number of heteroatoms can be 1 or 2; (16) In ring F, the unsubstituted or one or more R e-2 The number of alkene bonds in the substituted 4-12 alkene heterocycle is 1 or 2; (17) Each R P2 R e-1 and R e-2 In this context, the halogen is independently fluorine, chlorine, bromine, or iodine, such as fluorine; (18) Each R e-1 and R e-2 In, the unsubstituted C 1-6 The alkyl group is independently methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, or tert-butyl, for example, methyl or ethyl; (19) Each of the "multiples" refers to 2 or 3, for example, 2.

3. The compound of formula I as claimed in claim 2, characterized in that, It meets one or more of the following conditions: (1)L 2 In, the unsubstituted or unreplaced R P2 The 5-10-membered heteroaryl group in the substituted 5-10-membered heteroaryl group is independently a 5-6-membered heteroaryl group, wherein the heteroatom of the 5-6-membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2, or 3, such as pyridyl, pyrimidinyl, pyrazinyl, or pyridazinyl, for example... (2)L 3 In, the unsubstituted or unreplaced R P2 The 3-12-membered heterocyclic alkyl group in the substituted 3-12-membered heterocyclic alkyl group is independently a 3-6-membered monocyclic heterocyclic alkyl group, wherein the heteroatom of the 3-6-membered monocyclic heterocyclic alkyl group is independently selected from one or both of N and O, and the number of heteroatoms is independently one or two; for example, piperazine group, and another example... (3) In ring F, the unsubstituted or unreplaced R e-2 The 4-12-membered olefinic heterocycle is replaced with a 5-6-membered olefinic heterocycle, wherein the heteroatom of the 5-6-membered olefinic heterocycle is N, the number of heteroatoms is 1 or 2, and the number of olefin bonds is 1 or 2, for example...

4. The compound of formula I as claimed in claim 1, characterized in that, It meets one or more of the following conditions: (1)L 1 For not replaced or by an R a Replacement C 2-6 alkynyl group; (2) Each R a Independently for a R a1 Substituted 6-10 aryl groups; (3) Each R a1 Independently for C substituted with one hydroxyl group 1-6 alkyl; (4)L 2 For not replaced or by one or more R P2 Substituted phenylene or unsubstituted or with one or more R P2 The substituted 5-6-membered heteroaryl group; the heteroatom of the 5-6-membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2, or 3; preferably, L 2 It is an unsubstituted phenylene or an unsubstituted 5-6 membered heteroaryl group; the heteroatom of the 5-6 membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2 or 3; (5)L 3 For not replaced or by one or more R P2 The substituted 3-6-membered monocyclic heterocyclic alkylene group, wherein the heteroatoms of the 3-6-membered monocyclic heterocyclic alkylene group are independently selected from one or both of N and O, and the number of heteroatoms is independently one or two; preferably, L 3 It is an unsubstituted 3-6 membered monocyclic heterocyclic alkylene group, wherein the heteroatom of the 3-6 membered monocyclic heterocyclic alkylene group is N, and the number of heteroatoms is 1 or 2; (6)L 4 For not replaced or by one or more R P2 Replacement C 1-8 Alkylene; preferably, L 4 For unreplaced C 1-8 Alkylene; (7) Ring E is unsubstituted or replaced by one or more R e-1 Substituted benzene ring or unsubstituted or with one or more R e-1 The substituted 5-6 membered heteroaromatic ring, wherein the heteroatom in the 5-6 membered heteroaromatic ring is N, and the number of heteroatoms is 1 or 2; (8) Each R e-1 Halogens are independent of each other; (9) Ring F is unsubstituted or is replaced by one or more R e-2 The substituted 5-6 membered olefinic heterocycle, wherein the heteroatom of the 5-6 membered olefinic heterocycle is N, the number of heteroatoms is one or two, and the number of olefin bonds is one or two; preferably, the ring F is substituted by one or more R e-2 Substituted 5-6 membered olefinic heterocycles; (10) Each R e-2 Independent of an oxo group or an unsubstituted C 1-6 alkyl.

5. The compound of formula I as claimed in claim 1, characterized in that, It meets one or more of the following conditions: (1)L 1 for (2)L 2 for For example For example Ideally, L 2 for 1 bit and L 2 Connection, 2 bits and L 3 Connection; for example 1 bit and L 2 Connection, 2 bits and L 3 Connection; for example 1 bit and L 2 Connection, 2 bits and L 3 connect; (3)L 3 for (4)L 4 for (5) for The better location is 6. The compound of formula I as claimed in claim 1, characterized in that, L 1 For not replaced or by an R a Replacement C 2-6 alkynyl group; R a For being an R a1 Substituted 6-10 aryl groups; R a1 C replaced by a hydroxyl group 1-6 alkyl; L 2 For not replaced or by one or more R P2 Substituted phenylene or unsubstituted or with one or more R P2 The substituted 5-6-membered heteroaryl group; wherein the heteroatom of the 5-6-membered heteroaryl group is N, and the number of heteroatoms is independently 1, 2 or 3; L 3 It is an unsubstituted 3-6 membered monocyclic heterocyclic alkylene group, wherein the heteroatom of the 3-6 membered monocyclic heterocyclic alkylene group is N, and the number of heteroatoms is 1 or 2; L 4 For unreplaced C 1-8 Alkylene; Ring E is unsubstituted or replaced by one or more R e-1 Substituted benzene ring or unsubstituted or with one or more R e-1 The substituted 5-6 membered heteroaromatic ring, wherein the heteroatom in the 5-6 membered heteroaromatic ring is N, and the number of heteroatoms is 1 or 2; Each R e-1 Halogens are independent of each other; Ring F is formed by one or more R e-2 A substituted 5-6 membered olefinic heterocycle; wherein the heteroatom of the 5-6 membered olefinic heterocycle is N, the number of heteroatoms is 1 or 2, and the number of olefin bonds is 1 or 2; Each R e-2 Independent of an oxo group or an unsubstituted C 1-6 alkyl.

7. The compound of formula I as claimed in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compound shown in Formula I is any of the following compounds:

8. A pharmaceutical composition, characterized in that, It includes: (1) The compound of formula I as described in any one of claims 1-7, or a pharmaceutically acceptable salt thereof, and (2) Pharmaceutical excipients.

9. The use of substance A in the preparation of a PARP1 enzyme inhibitor, wherein substance A is a compound of formula I as described in any one of claims 1-7, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 8.

10. The use of substance A in the preparation of a medicament for treating and / or preventing diseases associated with the PARP1 enzyme, wherein the diseases associated with the PARP1 enzyme may be breast cancer, colorectal cancer, prostate cancer, or pancreatic cancer, wherein substance A is a compound of formula I as described in any one of claims 1-7, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition as described in claim 8.

Images