Nitrogen-containing heterocyclic compound as well as preparation method and medical application thereof

Abstract

The invention relates to a nitrogen-containing heterocyclic compound as well as a preparation method and medical application thereof. In particular, the present invention relates to a compound represented by general formula (I), a preparation method thereof, a pharmaceutical composition containing the compound, and a use thereof as a PARP1 inhibitor. The compound and the pharmaceutical composition containing the compound can be used for treating and / or preventing diseases related to PARP1 activity, such as cancers carrying BRCA gene mutation and the like. Wherein the definition of each group in the general formula (I) is the same as that in the specification. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and specifically relates to nitrogen-containing heterocyclic compounds, preparation methods thereof and pharmaceutical compositions containing the same, as well as use of the nitrogen-containing heterocyclic compounds as PARP1 inhibitors in the treatment and / or prevention of diseases associated with PARP1 activity. Background Art

[0002] Poly (ADP-ribose) polymerases (PARPs) play an important role in DNA replication, recombination, chromatin remodeling and DNA damage repair. The family includes 18 members with ADP-ribosyltransferase activity, catalyzing the addition of ADP-ribose units to DNA or different receptor proteins, affecting a variety of cellular processes. PARP1 and PARP2 are the most widely studied PARPs because of their role in DNA damage repair. PARP1 is the most important member of the PARP family. It is a nuclear protein composed of three domains: a DNA binding domain containing two zinc fingers at the N-terminus, a self-modification domain and a C-terminal catalytic domain. It undertakes more than 90% of the functions of the PARPs family in cells and is a key factor in DNA damage repair. PARP2 has similar functions to PARP1, but the two differ in substrate selection. PARP1 and PARP2 can repair DNA single-strand breaks (SSBs), and PARP1 can also repair DNA double-strand breaks (DSBs) and replication fork damage. PARP1 is a sensor for DNA breaks. It is activated after DNA damage and recognizes and binds to DNA breaks through its zinc finger domain, reducing recombination and protecting damaged DNA from the effects of exonucleases. After binding to the DNA gap, PARP1 catalyzes the glycosylation of nicotinamide adenine dinucleotide (NAD +) is decomposed into nicotinamide and ADP ribose, and then ADP ribose is used as a substrate to make the receptor protein and PARP1 itself "PARylated", forming PARP-ADP-ribose branched chains, which can prevent the nearby DNA molecules from recombination with the damaged DNA, and on the other hand, it can attract DNA repair proteins to bind and reduce the affinity of PARP1 with DNA, so that PARP1 dissociates from the DNA break, and then DNA repair proteins bind to the DNA gap to repair the damaged site. The "PARylation" of PARP1 will be cleared by other enzymes, so that PARP1 can regain activity and look for the next DNA break. In eukaryotic cells, there is not only the formation of PAR chains, but also the degradation of PAR chains catalyzed by poly (ADP-ribose) glycohydrolase [PARG] and type 3 ADP ribosylhydrolase (ADP-ribosylhydrolase 3, ARH3). Four enzymes, PARP1, PARP2, PARP5A and PARP5B, can catalyze the synthesis of PAR chains. Most other enzymes in the superfamily can only catalyze the synthesis of a single ADP-ribose unit, so they are also called mono(ADP-ribosyl)ases (MARs).

[0003] The binding of PARP to DNA damage sites, its catalytic activity, and its release from DNA are all important steps in the tumor cell response to DNA damage caused by chemotherapeutic drugs and radiation. Inhibition of PARP family enzymes has been used as a strategy to selectively kill tumor cells by inhibiting DNA repair pathways. Small molecule PARP1 inhibitors have been shown to sensitize tumor cells to cytotoxic treatments (e.g., temozolomide, platinum, topoisomerase inhibitors, and radiation). A large number of preclinical and clinical studies have shown that tumor cells with deleterious BRCA1 or BRCA2 mutations are sensitive to PARP inhibitors. Tumors generated by mutation carriers usually lose the wild-type allele, which leads to tumor-specific homologous recombination repair (HRR) dysfunction at double-strand breaks, and thus rely on PARP function for survival. When PARP1 is inhibited, base excision repair is reduced, single-strand breaks generated during the normal cell cycle persist, and double-strand breaks can be formed when the replication fork encounters unrepaired breaks. Therefore, compared with wild-type cells, tumor cells lacking homologous recombination repair, such as BRCA1 and BRCA2 mutants, are highly sensitive to PARP inhibition. Currently, PARP inhibitor therapy is mainly targeted at cancer patients with BRCA mutations. However, for cancers with homologous recombination repair defects, each of the related genes may become a potential target for synthetic lethality of PARP inhibitors, and PARP inhibitors have great therapeutic value. For example, ATM deletions have been found in patients with T-lymphocytic leukemia, B-cell chronic lymphocytic leukemia, and breast cancer, CHK2 germline mutations have been found in sarcomas, breast cancer, ovarian cancer, and brain tumors, FANCC and FANCG mutations have been confirmed in pancreatic cancer, and FANCF promoter methylation has also occurred in ovarian cancer, breast cancer, cervical cancer, and lung cancer. Other homologous recombination repair-related genes have also been shown to constitute synthetic lethality with PARP. Homologous recombination repair is a multifactorial process that ensures the repair of DNA double bond breaks at damaged replication forks. PARP1 recruits MRE11 to the replication fork to promote base excision, PARP1 / 2 binds to 5'-deoxyribonucleic acid, and after PARP activation, it uses DNA polymerase β (Polβ) and ligase III to recruit the BER scaffold protein XRCC1. Cells with defects in XRCC1 and Polβ are highly sensitive to PARPis. PARP-DNA complex is a barrier to replication. In response to replication damage and stress, ataxia telangiectasia and rad3-related protein kinase (ATR) and cycle checkpoint kinase 1 (CHK1) activate the S phase checkpoint, slow down replication forks and block replication initiation, maintain genome stability, and inhibit ATR or CHK1, resulting in loss of S phase checkpoint, replication initiation, and double bond breaks. Combined use with PARP inhibitors shows synergistic effects. FANCI-FANCD2 can stabilize the RAD51-DNA complex, and PALB2 binds to BRCA2, promoting BRCA2 nuclear stability and accumulation.BRCA1 interacts with many HRR-associated factors (BARD1, CtIP, RAD51, BRCA2, PALB2, Abraxas, and RAP80) and accelerates HRR at multiple steps. The BRD family and BET proteins recognize and recruit multiple proteins to chromatin and transcription sites, as well as bind to acetylated histones. BET / BRD4 inhibitors inhibit HRR by blocking the transcription of DNA repair genes (such as CtIP, BRCA1, WEE1, TOPBP1, and RAD51), and show synergy with PARPis in cells with normal BRCA.

[0004] PARP1 / 2 inhibitors currently on the market or in clinical trials have severe hematological toxicity as monotherapy drugs, which limits chronic treatment plans or combination therapies. Therefore, it is necessary to develop compounds with high selectivity for PARP1 and less toxicity to meet clinical needs. Summary of the invention

[0005] After intensive research, the inventors designed and synthesized a series of nitrogen-containing heterocyclic compounds and screened them for PARP1 activity. The results showed that the compounds are potent and selective inhibitors, and have a selective inhibitory effect on BRCA-deficient cells. The compounds and pharmaceutical compositions containing the compounds can be used to treat and / or prevent diseases related to PARP1 activity, such as cancers carrying BRCA gene mutations.

[0006] Therefore, the object of the present invention is to provide a compound represented by general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer, deuterated substance, solvate, mixture thereof, or pharmaceutically acceptable salt,

[0007]

[0008] in:

[0009] is a single bond or a double bond;

[0010] A 1 and A 2 are each independently selected from N, C, O or CH;

[0011] X 1 , X 2 and X 3 are each independently selected from C or N;

[0012] B 1 and B 2 Each independently selected from O, N, S, NR 5 or CR 5 ;

[0013] L1 is selected from a bond, a carbonyl group or a CR 6 R 7 ;

[0014] L2 is selected from a bond, O, S, NR 6 or CR 6 R 7 ;

[0015] R 1 and R 2 are each independently selected from hydrogen, deuterium, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 3-5 Heterocyclic group, wherein the C 3-6 Cycloalkyl, C 3-5 The heterocyclic group is optionally substituted by 1, 2 or 3 R a Replace, or R 1 and R 2 Each does not exist;

[0016] R a is selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, mercapto, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted with one or more substituents selected from the group consisting of deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0017] Or, R 1 With R 2 , A 1 , A 2 Together they form C 3-7 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 aromatic ring or 5-14 membered heteroaromatic ring, wherein the C 3-7 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 The aromatic ring or 5-14 membered heteroaromatic ring is optionally replaced by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 Substitution of cycloalkyl groups;

[0018] Ring B is selected from a 4-6 membered saturated monocyclic heterocycle containing 1-2 nitrogen atoms, a 5-6 membered partially unsaturated monocyclic heterocycle containing 1-2 nitrogen atoms, a 6-8 membered saturated heterocyclic bridge ring containing 1-4 nitrogen atoms, a 5-10 membered saturated heterocyclic cyclopentadiene containing 1-4 nitrogen atoms, or a 5-11 membered saturated heterocyclic spirocycle containing 1-4 nitrogen atoms;

[0019] m is 0, 1, 2 or 3;

[0020] Each R 3 Each independently selected from deuterium, halogen, amino, cyano, hydroxyl, C 1-6 Alkyl, C 1-6 Cycloalkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkyl, deuterated C 1-4 Alkoxy; or two R on the same atom 3 Together with the carbon atom to which it is attached, it forms C=O;

[0021] Ring C is selected from a 5-6 membered monocyclic aromatic heterocycle containing 1-2 nitrogen, oxygen, or sulfur atoms, an 8-10 membered bicyclic aromatic heterocycle containing 1-2 nitrogen, oxygen, or sulfur atoms, and a 6-10 membered aromatic ring;

[0022] n is 0, 1, 2 or 3;

[0023] Each R 4 Each independently selected from deuterium, halogen, amino, amide, C(O)NR 4a R 4b , cyano, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkyl, deuterated C 1-4 Alkoxy;

[0024] R 4a and R 4b Each is selected from hydrogen, deuterium, C 1-6 Alkyl, C 3-6 Cycloalkyl, heterocyclyl, wherein the alkyl, cycloalkyl, heterocyclyl is optionally further substituted with one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0025] R 5Selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, mercapto, C 1-6 Alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted with one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

[0026] R 6 and R 7 are each independently selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; or

[0027] R 6 and R 7 The bond to which it is attached, C, N, O or S together form a cycloalkyl group, a heterocyclic group, and the cyclic group is substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl.

[0028] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, A 1 and A 2 are C and CH, CH and C, C and N, N and C, CH and O, O and CH or C and C respectively.

[0029] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, X 1 C or N, X 2 and X 3 For C.

[0030] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, B 1 and B 2 Each is NR 5 and CR 5 , CR 5 and NR 5 , O and CR5 , CR 5 and O, S and CR 5 , CR 5 and S, N and S, S and N, N and CR 5 , CR 5 and N or CR 5 and CR 5 .

[0031] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, L1 is CH2.

[0032] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, L2 is selected from a bond and O.

[0033] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, R 1 and R 2 are each independently selected from hydrogen and C 1-6 Alkyl, or R 1 and R 2 Each does not exist;

[0034] Or, R 1 With R 2 , A 1 , A 2 Together they form C 3-6 hydrocarbon ring, 3-6 membered heterocyclic ring containing 1-2 nitrogen, oxygen, sulfur atoms, C 6-10 aromatic ring or a 5-6 membered heteroaromatic ring containing 1-2 nitrogen, oxygen or sulfur atoms, wherein the hydrocarbon ring, heterocyclic ring, aromatic ring or heteroaromatic ring is optionally replaced by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 The cycloalkyl group is substituted with a substituent.

[0035] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomorph, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, ring B is selected from a 4-6 membered saturated monocyclic heterocycle containing 1-2 nitrogen atoms and a 5-6 membered partially unsaturated monocyclic heterocycle containing 1-2 nitrogen atoms; m is 0, and R 3 Does not exist.

[0036] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, ring B is selected from

[0037]

[0038] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomorph, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, ring C is selected from a 5-6 membered monocyclic aromatic heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms and an 8-10 membered bicyclic aromatic heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms; n is 0, 1 or 2; each R 4 are each independently selected from halogen, C(O)NR 4a R 4b and C 1-6 Alkyl; R 4a and R 4b are each independently selected from hydrogen, C 1-6 Alkyl and deuterated C 1-6 alkyl.

[0039] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, Selected from

[0040] In a preferred embodiment, in the compound represented by the above general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, R 5 Selected from hydrogen and C 1-6 alkyl.

[0041] In a preferred embodiment, the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt is a compound represented by general formula (II) or its tautomer, mesomer, racemate, enantiomer, diastereomer, or pharmaceutically acceptable salt thereof:

[0042]

[0043] Among them, ring A is C 3-6 hydrocarbon ring, 3-6 membered heterocyclic ring containing 1-2 nitrogen, oxygen, sulfur atoms, C 6-10aromatic ring or a 5-6 membered heteroaromatic ring containing 1-2 nitrogen, oxygen or sulfur atoms, wherein the hydrocarbon ring, heterocyclic ring, aromatic ring or heteroaromatic ring is optionally replaced by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 Substitution of cycloalkyl groups;

[0044] A 1 , A 2 , X 1 , X 2 , X 3 , B 1 , B 2 , L1, L2, R 3 , R 4 , Ring B, Ring C, m and n are as defined in Formula (I).

[0045] In a preferred embodiment, in the compound represented by the above general formula (II) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, ring A is selected from wherein the ring A is optionally substituted by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 The cycloalkyl group is substituted with a substituent.

[0046] In a preferred embodiment, in the compound represented by the above general formula (II) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, ring A is selected from

[0047]

[0048] In a preferred embodiment, the compound represented by the general formula (I) above or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt is a compound represented by the general formula (I-1) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt:

[0049]

[0050] in

[0051] X 1 C or N;

[0052] A 1 , A 2 , R 1 , R 2 , B 1 , B 2, L2, R 4 , Ring B, Ring C and n are as defined in formula (I).

[0053] In a preferred embodiment, the compound represented by the general formula (I-1) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt is a compound represented by the general formula (I-2) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt:

[0054]

[0055] in

[0056] Ring A is as defined in formula (II);

[0057] X 1 As defined in formula (I-1);

[0058] A 1 , A 2 , B 1 , B 2 , L2, R 4 , Ring B, Ring C and n are as defined in Formula (I). Typical compounds of the present invention include but are not limited to:

[0059]

[0060]

[0061]

[0062]

[0063]

[0064]

[0065]

[0066]

[0067]

[0068]

[0069]

[0070] Its tautomer, meso racemate, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt.

[0071] Another aspect of the present invention provides a method for preparing the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, which comprises the following steps:

[0072]

[0073] The compound represented by the general formula III or its salt (preferably hydrochloride) undergoes an affinity substitution reaction with the compound represented by the general formula IV under alkaline conditions, optionally in the presence of a catalyst, to obtain a compound represented by the general formula I or a pharmaceutically acceptable salt thereof;

[0074] Wherein Z is a halogen, preferably a chlorine atom;

[0075] A 1 , A 2 , X 1 , X 2 , X 3 , B 1 , B 2 , L1, L2, Ring B, Ring C, m and n are as defined in Formula (I).

[0076] The base includes organic base and inorganic base. The catalyst of the affinity substitution reaction is sodium iodide or potassium iodide.

[0077] Another aspect of the present invention provides a method for preparing the compound represented by general formula (II) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, which comprises the following steps:

[0078]

[0079] The compound represented by the general formula V or its salt (preferably hydrochloride) undergoes an affinity substitution reaction with the compound represented by the general formula IV under alkaline conditions, optionally in the presence of a catalyst, to obtain a compound represented by the general formula II or a pharmaceutically acceptable salt thereof;

[0080] Wherein Z is a halogen, preferably a chlorine atom;

[0081] A 1 , A 2 , X 1 , X 2 , X 3 , B 1 , B 2 , L1, L2, Ring A, Ring B, Ring C, m and n are as defined in Formula (II).

[0082] The base includes organic base and inorganic base. The catalyst of the affinity substitution reaction is sodium iodide or potassium iodide.

[0083] Another aspect of the present invention provides a pharmaceutical composition comprising the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0084] The present invention further provides use of the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, in the preparation of a PARP1 inhibitor.

[0085] The present invention further provides use of the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, in the preparation of a medicament for preventing and / or treating diseases associated with PARP1 activity.

[0086] The present invention further provides the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt or a pharmaceutical composition containing the same, for use as a drug.

[0087] The present invention further provides a compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, which is used as a PARP1 (poly (ADP-ribose) polymerase 1) inhibitor.

[0088] The present invention further provides the compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or its pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, for use in preventing and / or treating diseases associated with PARP1 activity.

[0089] The present invention further provides a method for inhibiting PARP1 activity, which comprises administering to a subject in need thereof an effective amount of a compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same.

[0090] The present invention further provides a method for preventing and / or treating diseases associated with PARP1 activity, comprising administering to a subject in need thereof a preventively or therapeutically effective amount of a compound according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same.

[0091] In a preferred embodiment of the present invention, the disease associated with PARP1 activity according to the present invention may be a solid tumor, such as ovarian cancer, breast cancer, prostate cancer, and the like.

[0092] The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, such as tablets, troches, lozenges, aqueous or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs. Oral compositions may be prepared according to any method known in the art for preparing pharmaceutical compositions, and such compositions may contain one or more ingredients selected from the following: sweeteners, flavoring agents, colorants and preservatives to provide pleasing and palatable pharmaceutical preparations. Tablets contain the active ingredient and non-toxic pharmaceutically acceptable excipients suitable for preparing tablets for mixing. These excipients may be inert excipients such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating agents and disintegrants such as microcrystalline cellulose, crosslinked sodium carboxymethyl cellulose, corn starch or alginic acid; binders such as starch, gelatin, polyvinyl pyrrolidone or gum arabic; and lubricants such as magnesium stearate, stearic acid or talc. These tablets may be uncoated or may be coated by known techniques which mask the taste of the drug or delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained release action over a longer period of time. For example, water soluble taste masking materials such as hydroxypropylmethylcellulose or hydroxypropylcellulose, or time extending materials such as ethylcellulose, cellulose acetate butyrate may be used.

[0093] Oral preparations may also be provided in hard gelatin capsules wherein the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or in soft gelatin capsules wherein the active ingredient is mixed with a water-soluble carrier such as polyethylene glycol or an oily vehicle such as peanut oil, liquid paraffin or olive oil.

[0094] Aqueous suspensions contain the active substance and excipients suitable for preparing aqueous suspensions for mixing. Such excipients are suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone and gum arabic; dispersants or wetting agents, which may be naturally occurring phosphatides such as lecithin, or condensation products of alkylene oxides with fatty acids, for example, polyoxyethylene stearate, or condensation products of ethylene oxide with long chain fatty alcohols, for example, heptadecaethyleneoxy cetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol, for example, polyethylene oxide sorbitan monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyethylene oxide dehydrated sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, for example ethylparaben or n-propylparaben, one or more coloring agents, one or more flavoring agents and one or more sweetening agents, for example sucrose, saccharin or aspartame.

[0095] Oil suspensions can be prepared by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. The oil suspension may contain a thickener such as beeswax, hard paraffin or cetyl alcohol. The above-mentioned sweeteners and flavoring agents may be added to provide a palatable preparation. These compositions may be preserved by adding an antioxidant such as butylated hydroxyanisole or alpha-tocopherol.

[0096] Dispersible powders and granules suitable for preparing aqueous suspensions can provide the active ingredient and a dispersant or wetting agent, a suspending agent or one or more preservatives for mixing by adding water. Suitable dispersants or wetting agents and suspending agents are as described above. Other excipients such as sweeteners, flavoring agents and coloring agents can also be added. These compositions can be preserved by adding antioxidants such as ascorbic acid.

[0097] The pharmaceutical composition of the present invention can also be in the form of an oil-in-water emulsion. The oil phase can be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin or a mixture thereof. Suitable emulsifiers can be naturally occurring phospholipids, such as soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and condensation products of the partial esters and ethylene oxide, such as polyethylene oxide sorbitol monooleate. Emulsions can also contain sweeteners, flavoring agents, preservatives, and antioxidants. Syrups and elixirs prepared with sweeteners such as glycerol, propylene glycol, sorbitol, or sucrose can be used. Such preparations can also contain a demulcent, a preservative, a coloring agent, and an antioxidant.

[0098] The pharmaceutical composition of the present invention may be in the form of a sterile injectable aqueous solution. Acceptable vehicles and solvents that may be used are water, Ringer's solution and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in an oil phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then added to a mixture of water and glycerol and processed to form a microemulsion. The injection or microemulsion may be injected into the patient's bloodstream by local mass injection. Alternatively, the solution and microemulsion may be preferably administered in a manner that maintains a constant circulating concentration of the compound of the present invention. To maintain this constant concentration, a continuous intravenous drug delivery device may be used.

[0099] The pharmaceutical composition of the present invention can be in the form of a sterile injection water or oil suspension for intramuscular and subcutaneous administration. The suspension can be prepared according to known techniques with the above-mentioned suitable dispersants or wetting agents and suspending agents. The sterile injection preparation can also be a sterile injection solution or suspension prepared in a non-toxic parenterally acceptable diluent or solvent, such as a solution prepared in 1,3-butanediol. In addition, sterile fixed oils can be conveniently used as solvents or suspension media. For this purpose, any blended fixed oil including synthetic mono- or diglycerides can be used. In addition, fatty acids such as oleic acid can also be used to prepare injections.

[0100] The compounds of the invention may be administered in the form of suppositories for rectal administration. These pharmaceutical compositions may be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid in the rectum and which will dissolve in the rectum to release the drug. Such materials include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycol.

[0101] It is well known to those skilled in the art that the dosage of a drug depends on a variety of factors, including but not limited to the following factors: the activity of the specific compound used, the patient's age, the patient's weight, the patient's health condition, the patient's behavior, the patient's diet, the administration time, the administration method, the excretion rate, the combination of drugs, etc. In addition, the best treatment method, such as the mode of treatment, the daily dosage of the general compound or the type of pharmaceutically acceptable salt can be verified according to traditional treatment plans.

[0102] The present invention may contain the compound represented by the general formula (I), and a pharmaceutically acceptable salt, hydrate or solvate thereof as an active ingredient, mixed with a pharmaceutically acceptable carrier or excipient to prepare a composition, and prepared into a clinically acceptable dosage form. The derivatives of the present invention may be used in combination with other active ingredients, as long as they do not produce other adverse effects, such as allergic reactions, etc. The compounds of the present invention may be used as the sole active ingredient, or in combination with other drugs for treating diseases associated with PARP1 activity. Combination therapy is achieved by administering the individual therapeutic components simultaneously, separately or sequentially.

[0103] Definition of terms

[0104] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0105] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, an alkyl group containing 1 to 4 carbon atoms or an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, and the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0106] The term "alkylene" refers to a divalent alkyl group, wherein alkyl is as defined above, having from 1 to 20 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20) carbon atoms (i.e., C 1-20 The alkylene group is preferably an alkylene group having 1 to 12 carbon atoms (i.e., C1-12 alkylene), more preferably an alkylene group containing 1 to 6 carbon atoms (i.e., C 1-6 Alkylene), preferably an alkylene containing 1 to 4 carbon atoms (i.e., C 1-6 Alkylene). Non-limiting examples of alkylene include, but are not limited to, methylene (-CH2-), 1,1-ethylene (-CH(CH3)-), 1,2-ethylene (-CH2CH2)-, 1,1-propylene (-CH(CH2CH3)-), 1,2-propylene (-CH2CH(CH3)-), 1,3-propylene (-CH2CH2CH2-), and 1,4-butylene (-CH2CH2CH2CH2-). Alkylene may be substituted or unsubstituted, and when substituted, it may be substituted at any available point of attachment, and the substituent may be selected from one or more of alkyl, alkenyl, alkynyl, alkoxy, haloalkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, and oxo.

[0107] The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, preferably an alkenyl group containing 2 to 4 carbon atoms, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc. The alkenyl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0108] The term "alkynyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon triple bond, preferably an alkynyl group containing 2 to 4 carbon atoms or preferably an alkynyl group containing 3 to 4 carbon atoms, such as ethynyl, propynyl, butynyl, etc. Alkynyl groups may be substituted or unsubstituted, and when substituted, the substituents may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

[0109] The term "cycloalkyl" refers to a saturated or partially unsaturated (i.e., not having a completely conjugated π electron system) monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring comprising 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include cycloalkyls of spirocyclic, condensed and bridged rings.

[0110] The term "spirocycloalkyl" refers to a polycyclic group that shares a carbon atom (called a spiral atom) between 5 to 20 monocyclic rings, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, more preferably 7 to 10 yuan. According to the number of spiral atoms shared between the rings, the spirocycloalkyl is divided into a single spiral cycloalkyl, a double spiral cycloalkyl or a multi-spirocycloalkyl, preferably a single spiral cycloalkyl and a double spiral cycloalkyl. More preferably, it is a 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral cycloalkyl. Non-limiting examples of spirocycloalkyl include:

[0111]

[0112] The term "condensed cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic condensed cycloalkyl, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic alkyl. Non-limiting examples of condensed cycloalkyls include:

[0113]

[0114] The term "bridged cycloalkyl" refers to a 5 to 20-membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 members, and more preferably 7 to 10 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably a bicyclic, tricyclic or tetracyclic, and more preferably a bicyclic or tricyclic. Non-limiting examples of bridged cycloalkyl include:

[0115]

[0116] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, tetrahydrobenzofuranyl, tetrahydrobenzoxazolyl, tetrahydrobenzisoxazolyl, cyclopentathienyl, tetrahydrobenzothiazolyl, etc. The cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0117] The term "heterocyclyl" refers to a saturated or partially unsaturated (i.e., not having a completely conjugated π-electron system) monocyclic or polycyclic cyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, but does not include -OO-, -OS- or -SS- ring parts, and the remaining ring atoms are carbon. Preferably, it contains 4 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 7 to 12 ring atoms, of which 1 to 4 are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably 1, 2, 5-oxadiazolyl, pyranyl or morpholinyl. Polycyclic heterocyclic groups include spirocyclic, condensed ring and bridged heterocyclic groups.

[0118] The term "spiro heterocyclic group" refers to a polycyclic heterocyclic group in which the monocyclic rings of 5 to 20 members share one atom (called a spiro atom), wherein one or more of the ring atoms is selected from nitrogen, oxygen or S(O) m (wherein m is an integer 0 to 2) heteroatom, and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated π electron system. Preferably, it is 6 to 14 yuan, more preferably 7 to 12 yuan. According to the number of shared spiral atoms between rings, the spiral heterocyclic group is divided into a single spiral heterocyclic group, a double spiral heterocyclic group or a multi-spiro heterocyclic group, preferably a single spiral heterocyclic group and a double spiral heterocyclic group. More preferably, it is a 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan single spiral heterocyclic group. Non-limiting examples of spiral heterocyclic groups include:

[0119]

[0120] The term "fused heterocyclyl" refers to a polycyclic heterocyclic group of 5 to 20 members, each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated π electron system, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon. Preferably 6 to 14 members, more preferably 7 to 12 members. According to the number of constituent rings, it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 5-membered / 5-membered or 5-membered / 6-membered bicyclic fused heterocyclic group. Non-limiting examples of fused heterocyclic groups include:

[0121] The term "bridged heterocyclyl" refers to a 5- to 14-membered polycyclic heterocyclic group in which any two rings share two atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a completely conjugated π-electron system, and one or more of the ring atoms is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, and the remaining ring atoms are carbon. Preferably, it is 6 to 14 members, and more preferably 7 to 12 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic. Non-limiting examples of bridged heterocyclic groups include:

[0122]

[0123] The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is the heterocyclyl.

[0124] The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0125] The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, such as phenyl and naphthyl. More preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, non-limiting examples of which include:

[0126]

[0127] The aryl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0128] The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. The heteroaryl group is preferably 5 to 10 members, containing 1 to 3 heteroatoms; more preferably 5 or 6 members, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidyl, thiadiazole, pyrazinyl, etc., preferably imidazolyl, thiazolyl, pyrazolyl or pyrimidyl, thiazolyl; more preferably pyrazolyl or thiazolyl. The heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, non-limiting examples of which include:

[0129]

[0130] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0131] The term "heteroalkyl" refers to a straight or branched chain alkyl group containing 1 to 20 carbon atoms and 1 to 3 heteroatoms selected from O, N, Si and S, wherein alkyl is as defined above, and wherein N and S may be optionally oxidized and N may be optionally quaternized.

[0132] The term "alkoxy" refers to-O-(alkyl), wherein the definition of alkyl is as described above. The non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or unsubstituted, and when substituted, substituents can be one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0133] The term "cycloalkoxy" refers to an -O-(cycloalkyl) group, wherein cycloalkyl is as defined above.

[0134] The term "heterocycloalkoxy" refers to -O-(heterocyclyl) wherein heterocyclyl is as defined above.

[0135] The term "cycloalkylthio" refers to -S-(cycloalkyl) wherein cycloalkyl is as defined above.

[0136] The term "heterocycloalkylthio" refers to -S-(heterocyclyl) wherein heterocyclyl is as defined above.

[0137] The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.

[0138] The term "haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

[0139] The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.

[0140] The term "hydroxy" refers to an -OH group.

[0141] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0142] The term "amino" refers to -NH2.

[0143] The term "cyano" refers to -CN.

[0144] The term "nitro" refers to -NO2.

[0145] The term "oxo" refers to =0.

[0146] The term "carboxy" refers to -C(O)OH.

[0147] The term "thiol" refers to -SH.

[0148] The term "ester group" refers to -C(O)O(alkyl) or -C(O)O(cycloalkyl), wherein alkyl and cycloalkyl are as defined above.

[0149] The term "acyl" refers to a compound containing a -C(O)R group, wherein R is alkyl, cycloalkyl, heterocyclyl, aryl, or heteroaryl.

[0150] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

[0151] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (e.g. olefinic) bonds.

[0152] "Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0153] "Pharmaceutically acceptable salts" or "pharmaceutically acceptable salts" refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.

[0154] "Carrier" refers to a carrier or diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound.

[0155] Synthesis method of the compound of the present invention

[0156] Another aspect of the present invention provides a method for preparing the compound represented by general formula (I) according to the present invention or its tautomer, mesomer, racemate, enantiomer, diastereomer, or mixture thereof, or pharmaceutically acceptable salt thereof, using the following technical scheme:

[0157] Solution 1

[0158]

[0159] The compound represented by the general formula III or its salt (preferably hydrochloride) undergoes an affinity substitution reaction with the compound represented by the general formula IV under alkaline conditions, optionally in the presence of a catalyst, to obtain a compound represented by the general formula I or a pharmaceutically acceptable salt thereof;

[0160] Wherein Z is a halogen, preferably a chlorine atom;

[0161] R 1 , R 2 , R 3 , R 4 , A 1 , A 2 , X 1 , X 2 , X 3, B 1 , B 2 , L1, L2, Ring B, Ring C, m and n are as defined in Formula (I).

[0162] The base includes organic bases and inorganic bases; the organic bases include but are not limited to triethylamine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, sodium acetate, potassium acetate, sodium tert-butoxide and potassium tert-butoxide, preferably N,N-diisopropylethylamine; the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, cesium carbonate, cesium fluoride, sodium hydroxide, lithium hydroxide monohydrate, lithium hydroxide and potassium hydroxide.

[0163] The catalyst for the affinity substitution reaction is sodium iodide or potassium iodide, preferably sodium iodide.

[0164] The reaction is preferably carried out in a solvent, and the solvent used includes but is not limited to: N-methylpyrrolidone, ethylene glycol dimethyl ether, acetic acid, methanol, ethanol, acetonitrile, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, water, N,N-dimethylformamide, N,N-dimethylacetamide, 1,2-dibromoethane, 1,2-dichloroethane and mixtures thereof.

[0165] Solution 2

[0166]

[0167] The compound represented by general formula V or its salt (preferably hydrochloride) undergoes an affinity substitution reaction with the compound represented by general formula IV under alkaline conditions, optionally in the presence of a catalyst, to obtain a compound represented by general formula II or a pharmaceutically acceptable salt thereof.

[0168] Wherein Z is a halogen, preferably a chlorine atom;

[0169] R 3 , R 4 , A 1 , A 2 , X 1 , X 2 , X 3 , B 1 , B 2 , L1, L2, Ring A, Ring B, Ring C, m and n are as defined in Formula (II).

[0170] The base includes organic bases and inorganic bases; the organic bases include but are not limited to triethylamine, N,N-diisopropylethylamine, n-butyllithium, lithium diisopropylamide, sodium acetate, potassium acetate, sodium tert-butoxide and potassium tert-butoxide, preferably N,N-diisopropylethylamine; the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, cesium carbonate, cesium fluoride, sodium hydroxide, lithium hydroxide monohydrate, lithium hydroxide and potassium hydroxide.

[0171] The catalyst for the affinity substitution reaction is sodium iodide or potassium iodide, preferably sodium iodide.

[0172] The reaction is preferably carried out in a solvent, and the solvent used includes but is not limited to: N-methylpyrrolidone, ethylene glycol dimethyl ether, acetic acid, methanol, ethanol, acetonitrile, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1,4-dioxane, water, N,N-dimethylformamide, N,N-dimethylacetamide, 1,2-dibromoethane, 1,2-dichloroethane and mixtures thereof. DETAILED DESCRIPTION

[0173] The present invention is further described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

[0174] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of (ppm) is given. NMR measurements were performed using a Bruker dps300 nuclear magnetic spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), deuterated methanol (CD3OD) as the measuring solvent, and tetramethylsilane (TMS) as the internal standard.

[0175] LC-MS was measured using a 1100 Series LC / MSD Trap (ESI) mass spectrometer (manufacturer: Agilent).

[0176] GC-MS measurement was performed using GCMS-QP2010 SE.

[0177] The preparative liquid chromatography method used LC3000 high performance liquid chromatograph and LC6000 high performance liquid chromatograph (manufacturer: Chuangxin Tongheng), and the chromatographic column was Daisogel C18 10 μm 60A (20 mm×250 mm).

[0178] High performance liquid chromatography (HPLC) was performed using Shimadzu LC-20AD high pressure liquid chromatograph (Agilent TC-C18 250×4.6 mm 5 μm column) and Shimadzu LC-2010AHT high pressure liquid chromatograph (Phenomenex C18 250×4.6 mm 5 μm column).

[0179] The thin layer chromatography silica gel plate used was Qingdao Ocean Chemical GF254 silica gel plate. The silica gel plate used in thin layer chromatography (TLC) had a specification of 0.15 mm to 0.2 mm, and the specification used for thin layer chromatography separation and purification products was 0.4 mm to 0.5 mm.

[0180] Column chromatography generally uses Qingdao marine silica gel 100-200 mesh and 200-300 mesh silica gel as the carrier.

[0181] The known starting materials of the present invention can be synthesized by methods known in the art, or can be purchased from online shopping malls, Beijing Coupling, Sigma, Bailingwei, Yishiming, Shanghai Shuya, Inokai, Nanjing Yaoshi, Anaiji Chemical and other companies.

[0182] Unless otherwise specified in the examples, the reactions can be carried out under an argon atmosphere or a nitrogen atmosphere.

[0183] Argon atmosphere or nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a capacity of about 1L.

[0184] A CEM Discover SP microwave reactor was used for the microwave reaction.

[0185] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0186] Unless otherwise specified in the examples, the reaction temperature is room temperature, particularly 20°C to 30°C.

[0187] The reaction progress in the embodiment is monitored by thin layer chromatography (TLC), and the developing solvent systems used in the reaction are: A: dichloromethane and methanol system, B: n-hexane and ethyl acetate system, C: petroleum ether and ethyl acetate system, D: acetone, and the volume ratio of the solvent is adjusted according to the polarity of the compound.

[0188] The eluent system of column chromatography and the developing solvent system of thin layer chromatography used for purifying compounds include: A: dichloromethane and methanol system, B: petroleum ether, ethyl acetate and dichloromethane system, C: petroleum ether and ethyl acetate system. The volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.

[0189] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the present invention.

[0190] Example 1: Preparation of 5-(4-((6-ethyl-5-oxo-4,5-dihydro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (1)

[0191]

[0192]

[0193] Step 1: Preparation of 2-methoxy-6-methyl-5-nitro-3-vinylpyridine (1a).

[0194] 3-Bromo-2-methoxy-6-methyl-5-nitropyridine (2.47 g, 10.0 mmol), potassium ethylene trifluoroborate (2.01 g, 15.0 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (363 mg, 0.50 mmol) and sodium carbonate (3.18 g, 30.0 mmol) were dissolved in dioxane (32 mL) and water (8 mL), and reacted at 90° C. for 4 hours under nitrogen protection. The reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 5) to obtain 1.52 g of the title compound as a light yellow solid, with a yield of 78.4%.

[0195] LC-MS: m / z = 195.2 [M+H] + .

[0196] Step 2: Preparation of (E)-2-(6-methoxy-3-nitro-5-vinylpyridin-2-yl)-N,N-dimethylethen-1-amine (1b).

[0197] 2-Methoxy-6-methyl-5-nitro-3-vinylpyridine (1.52 g, 7.83 mmol) and N,N-dimethylformamide dimethyl acetal (1.21 g, 10.2 mmol) were dissolved in ethanol (15 mL) and stirred at 80° C. for 4 hours. The reaction solution was concentrated and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 5) to obtain 1.26 g of the title compound as a red oil with a yield of 64.6%.

[0198] LC-MS: m / z = 250.3 [M+H] + .

[0199] Step 3: Preparation of 6-ethyl-5-methoxy-1H-pyrrolo[3,2-b]pyridine (1c).

[0200] Dissolve (E)-2-(6-methoxy-3-nitro-5-vinylpyridin-2-yl)-N,N-dimethylvinyl-1-amine (1.26 g, 5.05 mmol) in ethanol (15 mL), add 10% palladium on carbon (130 mg), and stir at room temperature under 1 atmosphere of hydrogen to react overnight. Filter, concentrate the filtrate, and separate and purify the residue by silica gel column chromatography (mobile phase EA / PE = 1 / 20-1 / 5) to obtain 740 mg of the title compound as a light yellow solid, with a yield of 83.1%.

[0201] LC-MS: m / z = 177.2 [M+H] + .

[0202] Step 4: Preparation of 6-ethyl-5-methoxy-1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine (1d).

[0203] 6-Ethyl-5-methoxy-1H-pyrrolo[3,2-b]pyridine (740 mg, 4.20 mmol) was dissolved in tetrahydrofuran (15 mL), sodium hydride (252 mg, 6.30 mmol) was slowly added at 0°C, stirred at room temperature for 30 minutes, benzenesulfonyl chloride (971 mg, 5.50 mmol) in tetrahydrofuran (5 mL) was added dropwise at 0°C, stirred at room temperature and reacted overnight. 30 mL of water was added for dilution, ethyl acetate was extracted (20 mL×3), the organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 2) to obtain 720 mg of the title compound as a light yellow solid, with a yield of 54.2%.

[0204] LC-MS: m / z=317.4[M+H] + .

[0205] Step 5: Preparation of 6-ethyl-5-methoxy-1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine-2-carbaldehyde (1e).

[0206] 6-Ethyl-5-methoxy-1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine (700 mg, 2.21 mmol) and tetramethylethylenediamine (143 mg, 6.63 mmol) were dissolved in tetrahydrofuran (15 mL), and a solution of lithium diisopropylamide in tetrahydrofuran (2.0 M, 1.70 mL, 3.40 mmol) was slowly added dropwise at -78°C, and the mixture was reacted at -78°C for 1 hour, and N,N-dimethylformamide (210 mg, 2.87 mmol) was slowly added dropwise, and the mixture was reacted at -60°C for 2 hours. Saturated aqueous ammonium chloride solution (20 mL) was added to quench the reaction, and the reaction solution was extracted with ethyl acetate (20 mL×3), the organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 2) to obtain 470 mg of the title compound as a light yellow solid, with a yield of 61.8%.

[0207] LC-MS: m / z=345.4[M+H] + .

[0208] Step 6: Preparation of 5-(4-((6-ethyl-5-methoxy-1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (1f).

[0209] 6-Ethyl-5-methoxy-1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridine-2-carboxaldehyde (470 mg, 1.36 mmol) and N-methyl-5-(piperazin-1-yl)picolinamide (329 mg, 1.50 mmol) were dissolved in dichloromethane (20 mL), 2 drops of glacial acetic acid were added, and the mixture was stirred at room temperature for 4 hours. Sodium triacetoxyborohydride (577 mg, 2.72 mmol) was added, and the mixture was stirred at room temperature overnight. The reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase MeOH / DCM=1 / 50-1 / 10) to obtain 364 mg of the title compound as a light yellow solid, with a yield of 48.8%.

[0210] LC-MS: m / z = 549.6 [M+H] + .

[0211] Step 7: Preparation of 5-(4-((6-ethyl-5-methoxy-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (1 g).

[0212] 5-(4-((6-ethyl-5-methoxy-1-(phenylsulfonyl)-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (360 mg, 0.656 mmol) was dissolved in tetrahydrofuran (10 mL), tetrabutylammonium fluoride (858 mg, 3.28 mmol) was added under ice bath, and stirred at 50°C for 2 hours. 30 mL of water was added for dilution, and ethyl acetate was extracted (15 mL×3). The organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase MeOH / DCM=1 / 50-1 / 10) to obtain 114 mg of the title compound as a light yellow solid, with a yield of 42.5%.

[0213] LC-MS: m / z = 409.5 [M+H] + .

[0214] Step 8: Preparation of 5-(4-((6-ethyl-5-oxo-4,5-dihydro-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (1).

[0215] 5-(4-((6-ethyl-5-methoxy-1H-pyrrolo[3,2-b]pyridin-2-yl)methyl)piperazine-1-yl)-N-methylpicolinamide (80.0 mg, 0.196 mmol) was dissolved in 48% hydrobromic acid (5 mL) and stirred at 110°C for 1 hour. The reaction solution was concentrated and the residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid, 30 min) to obtain 5.60 mg of the title compound as a white solid with a yield of 7.24%.

[0216] LC-MS: m / z=395.5[M+H] + .

[0217] 1 H NMR (400MHz, DMSO-d6) δ11.37(s,1H),8.44(d,J=5.1Hz,1H),8.32(d,J=2.9Hz,1H),7.87(d,J=8.7Hz,1H),7.46(d,J=9.0Hz, 2H), 6.17 (s, 1H), 4.40 (s, 2H), 4.05 (s, 4H), 3.15 (s, 4H), 2.78 (d, J = 4.8Hz, 3H), 2.44 (q, J = 7.5Hz, 2H), 1.11 (t, J = 7.4Hz, 3H).

[0218] Example 2: Preparation of 5-(4-((6-ethyl-5-oxo-4,5-dihydrofuro[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (2)

[0219]

[0220] Step 1: Preparation of tert-butyl (2-formylfuran-3-yl)carbamate (2a).

[0221] 3-Bromo-2-furaldehyde (2.00 g, 11.4 mmol), tert-butyl carbamate (1.61 g, 13.7 mmol), cesium carbonate (5.20 g, 16.0 mmol), Xantphos (1.32 g, 2.29 mmol) and Pd2(dba)3 (1.30 g, 1.14 mmol) were dissolved in toluene (40 mL) and stirred at 100°C for 16 hours under a nitrogen atmosphere. The mixture was filtered and the filtrate was concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate = 10%-35%) to obtain 1.81 g of the title compound as a yellow solid. Yield: 75.0%.

[0222] LC-MS: m / z=212[M+H] + .

[0223] Step 2: Preparation of (E)-ethyl 2-((3-((tert-butoxycarbonyl)amino)furan-2-yl)methylene)butanoate (2b).

[0224] At room temperature, ethyl 2-(triphenylphosphine)butyrate (3.53 g, 9.37 mmol) was added to a solution of tert-butyl (2-formylfuran-3-yl)carbamate (1.80 g, 8.52 mmol) in THF (18 mL), and stirred at 60°C for 2 hours. The reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate = 5%-15%) to obtain 2.32 g of the title compound as a yellow solid, yield: 88.0%.

[0225] LC-MS: m / z=310[M+H] + .

[0226] Step 3: Preparation of (E)-ethyl 2-((3-aminofuran-2-yl)methylene)butanoate (2c).

[0227] Ethyl (E)-2-((3-((tert-butoxycarbonyl)amino)furan-2-yl)methylene)butanoate (2.32 g, 7.50 mmol) was dissolved in THF (3 mL), tetrabutylammonium fluoride (15 mL, 1 M in THF) was added dropwise, and stirred at 70°C for 16 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 25%-45%) to obtain 951 mg of the title compound as a yellow solid, with a yield of 60.6%.

[0228] LC-MS: m / z = 210 [M+H] + .

[0229] Step 4: Preparation of 6-ethylfuro[3,2-b]pyridin-5(4H)-one (2d).

[0230] At room temperature, NaH (454 mg, 11.4 mmol, 60%) was added to a solution of ethyl (E)-2-((3-aminofuran-2-yl)methylene)butanoate (951 mg, 4.54 mmol) in THF (15 mL), and the mixture was stirred at 70°C for 2 hours. The reaction solution was slowly added dropwise to 250 mL of water, extracted with ethyl acetate (250 mL x 2), the organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase: DCM:MeOH=0%-7%) to obtain 350 mg of the title compound as a yellow solid, yield: 47.2%.

[0231] LC-MS: m / z=164[M+H] + .

[0232] Step 5: Preparation of 6-ethyl-5-oxo-4,5-dihydrofuro[3,2-b]pyridine-2-carbaldehyde (2e).

[0233] At -78°C, n-butyl lithium (1.72 mL, 4.29 mmol, 2.5 M in Hexane) was slowly added dropwise to a solution of 6-ethylfuro[3,2-b]pyridin-5(4H)-one (350 mg, 2.14 mmol) in tetrahydrofuran (6 mL), and stirred at -60°C for 0.5 hour. N,N-dimethylformamide (248 μL, 4.29 mmol) was slowly added dropwise at -78°C, and stirred at -40°C for 1 hour. Saturated ammonium chloride solution (0.5 mL) was slowly added dropwise to the reaction solution, and the solution was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: DCM:MeOH=0%-7%) to obtain 250 mg of the title compound as a yellow oil.

[0234] LC-MS: m / z = 192 [M+H] + .

[0235] Step 6: Preparation of 5-(4-((6-ethyl-5-oxo-4,5-dihydrofuro[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (2).

[0236] N-Methyl-5-(piperazin-1-yl)picolinamide (86.0 mg, 0.392 mmol), glacial acetic acid (112 μL, 1.96 mmol) and 6-ethyl-5-oxo-4,5-dihydrofuro[3,2-b]pyridine-2-carbaldehyde (250 mg, 1.3 mmol) were dissolved in dichloromethane (3.0 mL), stirred at room temperature for 0.5 hours, sodium triacetoxyborohydride (108 mg, 510 μmol) was added, and stirred at room temperature for 16 hours. The reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: MeOH:DCM=0%-7%), and then separated again by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 27.0 mg of the title compound as a white solid. Yield: 17.4%.

[0237] LC-MS: m / z=395.95 [M+H] + .

[0238] 1 H NMR(400MHz,Methanol-d4)δ8.30(s,1H),7.92(s,1H),7.72(s,1H),7.41(s,1H),6.63(s,1H), 3.92(s,2H),3.47(d,J=15.2Hz,4H),2.87(s,7H),2.59(q,J=7.6Hz,2H),1.22(t,J=7.2Hz,3H).

[0239] Example 3: Preparation of 5-(4-((3-ethyl 1-2-oxo-1,2-dihydrothieno[2,3-b]pyrazin-6-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (3)

[0240]

[0241] Step 1: Preparation of methyl 4-((1-ethoxy-2-methylpropyl)-5-nitrothiophene-2-carboxylate (3a).

[0242] At room temperature, potassium carbonate (2.26 g, 16.38 mmol) was added to a solution of methyl 4-chloro-5-nitrothiophene-2-carboxylate (1.20 g, 5.46 mmol) and ethyl 2-aminobutyrate hydrochloride (1.01 g, 6.06 mmol) in acetonitrile (1.0 mL), and the mixture was stirred at 80°C for 2 hours. The reaction solution was concentrated, and the residual compound was separated and purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate = 35%-55%) to obtain 1.45 g of the title compound as a yellow solid. Yield: 84.0%.

[0243] LC-MS: m / z=317[M+H] + .

[0244] Step 2: Preparation of methyl 2-ethyl-3-oxo-1,2,3,4-tetrahydrothieno[2,3-b]pyrazine-6-carboxylate (3b).

[0245] Reduced iron powder (883 mg, 15.8 mmol, 100 mesh) was added to a solution of methyl 4-((1-ethoxy-2-methylpropyl)-5-nitrothiophene-2-carboxylate (1.00 g, 3.16 mmol) in glacial acetic acid (10 mL) at room temperature, and the mixture was stirred at 80° C. for 2 hours. The reaction solution was concentrated, and the residual compound was separated and purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate=45%-65%) to obtain 640 mg of the title compound as a yellow solid, yield: 84.3%.

[0246] LC-MS: m / z=241[M+H] + .

[0247] Step 3: Preparation of methyl 2-ethyl-3-oxo-3,4-dihydrothieno[2,3-b]pyrazine-6-carboxylate (3c).

[0248] At room temperature, add manganese dioxide (1.16 g, 13.3 mmol) to a solution of methyl 2-ethyl-3-oxo-1,2,3,4-tetrahydrothieno[2,3-b]pyrazine-6-carboxylate (640 mg, 2.66 mmol) in 1,4-dioxane (18 mL), and stir at room temperature for 16 hours. Filter, concentrate the filtrate, and separate and purify the residual compound by silica gel column chromatography (mobile phase: dichloromethane:methanol=0%-10%) to obtain 181 mg of the title compound as a yellow solid, yield: 28.5%.

[0249] LC-MS: m / z=239[M+H] + .

[0250] Step 4: Preparation of 2-ethyl-6-(hydroxymethyl)thieno[2,3-b]pyrazin-3(4H)-one (3d).

[0251] To a solution of methyl 2-ethyl-3-oxo-3,4-dihydrothieno[2,3-b]pyrazine-6-carboxylate (160 mg, 672 μmol) in tetrahydrofuran (6 mL) was slowly added dropwise lithium aluminum hydride (430 μL, 1.07 mmol, 2.5 M in THF) at 0°C and stirred at 0°C for 1 hour. Sodium sulfate decahydrate (320 mg) was slowly added at 0°C to quench the reaction, stirred at room temperature for 30 minutes, filtered, and the filtrate was concentrated to obtain 206 mg of the title compound as a yellow solid.

[0252] LC-MS: m / z = 211 [M+H] + .

[0253] Step 5: Preparation of 2-ethyl-3-oxo-3,4-dihydrothieno[2,3-b]pyrazine-6-carbaldehyde (3e).

[0254] At room temperature, manganese dioxide (852 mg, 9.80 mmol) was added to a solution of 2-ethyl-6-(hydroxymethyl)thieno[2,3-b]pyrazine-3(4H)-one (206 mg, 980 μmol) in tetrahydrofuran (10 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was filtered and the filtrate was concentrated. The residual compound was separated and purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate = 50%-80%) to obtain 55.0 mg of the title compound as a yellow solid, with a yield of 27.0%.

[0255] LC-MS: m / z = 209 [M+H] + .

[0256] Step 6: Preparation of 5-(4-((3-ethyl 1-2-oxo-1,2-dihydrothieno[2,3-b]pyrazin-6-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (3).

[0257] N-methyl-5-(piperazin-1-yl)picolinamide (58.0 mg, 264 μmol), glacial acetic acid (76.0 μL, 1.32 mmol) and 2-ethyl-3-oxo-3,4-dihydrothieno[2,3-b]pyrazine-6-carboxaldehyde (55.0 mg, 264 μmol) were dissolved in dichloromethane (5 mL), stirred at room temperature for 30 minutes, sodium triacetoxyborohydride (73.0 mg, 343 μmol) was added, and stirred at room temperature for 2 hours. The reaction solution was concentrated, and the residual compound was separated and purified by silica gel column chromatography (mobile phase: MeOH:DCM=0%-10%), and then separated again by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 19 mg of the title compound as a white solid. Yield: 17.4%.

[0258] LC-MS: m / z=412.95[M+H] + .

[0259] 1 H NMR (400MHz, DMSO-d6) δ12.53(s,1H),8.40(q,J=4.8Hz,1H),8.27(d,J=2.8Hz,1H),7.82(d,J=8.8Hz,1H),7.39(dd,J=8.8,2.8Hz,1H),6. 91(s,1H),3.82(s,2H),3.35(d,J=4.8Hz,4H),2.77(d,J=4.8Hz,3H),2.73(q,J=7.6Hz,2H),2.61(t,J=5.2Hz,4H),1.17(t,J=7.6Hz,3H).

[0260] Example 4: Preparation of 5-(4-((3-ethyl-2-oxo-2,3-dihydro-1H-thieno[2,3-b][1,4]oxazin-6-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (4)

[0261]

[0262]

[0263] Step 1: Preparation of methyl 5-((1-ethoxy-1-oxobutan-2-yl)oxy)-4-nitrothiophene-2-carboxylate (4a).

[0264] Dissolve methyl 5-chloro-4-nitrothiophene-2-carboxylate (1.108 g, 5.0 mmol), ethyl 2-hydroxybutyrate (990 mg, 7.50 mmol) and cesium carbonate (3.25 mg, 10.00 mmol) in tetrahydrofuran (30 mL), stir at 50 ° C for 18 hours, filter, concentrate the filtrate, and separate the residue by silica gel column chromatography (ethyl acetate / petroleum ether: 5%-10%) to obtain 513 mg of the title compound as a light yellow oil with a yield of 32.3%.

[0265] Step 2: Preparation of methyl 3-ethyl-2-oxo-2,3-dihydro-1H-thieno[2,3-b][1,4]oxazine-6-carboxylate (4b).

[0266] Dissolve 5-((1-ethoxy-1-oxobutane-2-yl)oxy)-4-nitrothiophene-2-carboxylic acid methyl ester (513 mg, 1.62 mmol) and reduced iron powder (900 mg, 16.1 mmol) in glacial acetic acid (20 mL) and stir at 80°C for 4 hours. The reaction solution was concentrated, methanol (10 mL) and dichloromethane (10 mL) were added to the residue, stirred at room temperature for 0.5 hours, filtered, the filter cake was washed with dichloromethane (10 mL×3), the organic phase was concentrated, and the residue was separated by column chromatography (methanol / dichloromethane: 2%-3%) to obtain 270 mg of the title compound as a light brown solid with a yield of 69.2%.

[0267] LC-MS: m / z = 241.9 [M+H] + .

[0268] Step 3: Preparation of 3-ethyl-6-(hydroxymethyl)-1H-thieno[2,3-b][1,4]oxazin-2(3H)-one (4c).

[0269] 3-Ethyl-2-oxo-2,3-dihydro-1H-thieno[2,3-b][1,4]oxazine-6-carboxylic acid methyl ester (235 mg, 0.975 mmol) and tetrahydrofuran (10 mL) were added to the reaction bottle. Under nitrogen protection, a solution of diisobutylaluminum hydride in tetrahydrofuran (1.0 M, 4.88 mL, 4.88 mmol) was slowly added dropwise at -78 °C. The temperature was slowly raised to room temperature and the reaction was continued for 4 hours. Sodium sulfate decahydrate was added to quench the reaction, and the reaction was filtered and the filtrate was concentrated to obtain 247 mg of a crude product of the title compound as a light yellow solid.

[0270] LC-MS: m / z = 214.0 [M+H] + .

[0271] Step 4: Preparation of 3-ethyl-2-oxo-2,3-dihydro-1H-thieno[2,3-b][1,4]oxazine-6-carbaldehyde (4d).

[0272] 3-Ethyl-6-(hydroxymethyl)-1H-thieno[2,3-b][1,4]oxazine-2(3H)-one (247 mg, 1.16 mmol), manganese dioxide (870 mg, 10.0 mmol) and dichloromethane (30 mL) were added to a reaction flask and stirred at 30°C for 16 hours. The mixture was filtered and the filtrate was concentrated. The residue was separated by silica gel column chromatography (ethyl acetate / petroleum ether: 20%-33%-50%) to obtain 40 mg of the title compound as a light brown solid with a yield of 16.3%.

[0273] LC-MS: m / z = 211.9 [M+H] + .

[0274] Step 5: Preparation of 5-(4-((3-ethyl-2-oxo-2,3-dihydro-1H-thieno[2,3-b][1,4]oxazin-6-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (4).

[0275] N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride (50.0 mg, 0.226 mmol) and triethylamine (24 mg, 0.226 mmol) were dissolved in dichloroethane (4 mL), reacted at room temperature for 1 hour, pH was adjusted to 5-6 with glacial acetic acid, 3-ethyl-2-oxo-2,3-dihydro-1H-thieno[2,3-b][1,4]oxazine-6-carbaldehyde (40 mg, 0.189 mmol) was added, reacted at room temperature for 1 hour, sodium triacetoxyborohydride (80 mg, 0.379 mmol) was added, reacted at room temperature for 16 hours, the reaction solution was concentrated, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 17.0 mg of the title compound as a light yellow solid, with a yield of 21.6%.

[0276] LC-MS: m / z=416.0[M+H] + .

[0277] 1H-NMR (400MHz, DMSO-d6) δ10.72(s,1H),8.48(q,J=5.2Hz,1H),8.34(d,J=2.8Hz,1H),7.89(d,J=8.4Hz,1H),7.48(dd,J1=7.4Hz, J2=2.8Hz,1H),6.82(d,1H),4.66-4.69(m,1H),4.50(m,2H),3.47(m,2H),3.13(m,4H),1.81-1.88(m,2H),0.99(t,J=7.2Hz,3H).

[0278] Example 5: Preparation of 5-(4-((6-ethyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (5)

[0279]

[0280] Step 1: Preparation of 2-methoxy-5-nitro-3-vinylpyridine (5a).

[0281] 3-Bromo-2-methoxy-5-nitropyridine (4.00 g, 17.2 mmol), tributyl vinyl tin (8.18 g, 25.8 mmol) and tetrakis(triphenylphosphine)palladium (993 mg, 0.86 mmol) were dissolved in 40 mL of N,N-dimethylformamide and stirred at 90°C for 16 hours under nitrogen protection. 60 mL of saturated KF aqueous solution was added and stirred at room temperature for 2 hours. 60 mL of ethyl acetate was added for dilution, filtered, the filtrate was separated, the organic phase was concentrated, and separated and purified by silica gel column chromatography (mobile phase EA / PE = 1 / 20 -1 / 5) to obtain 2.80 g of the title compound as a light yellow solid with a yield of 90.4%.

[0282] LC-MS: m / z = 181.1 [M+H] + .

[0283] Step 2: Preparation of 5-ethyl-6-methoxypyridin-3-amine (5b).

[0284] 2-Methoxy-5-nitro-3-vinylpyridine (2.80 g, 17.6 mmol) and 300 mg of 10% palladium on carbon were added to 30 mL of ethanol and stirred overnight at room temperature under 1 atmosphere of hydrogen. The mixture was filtered and the filtrate was concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase EA / PE = 1 / 20 -1 / 5) to obtain 2.10 g of the title compound as a light yellow oil with a yield of 78.4%.

[0285] LC-MS: m / z = 153.2 [M+H]+ .

[0286] Step 3: Preparation of 6-ethyl-5-methoxythiazolo[5,4-b]pyridin-2-amine (5c).

[0287] Dissolve 5-ethyl-6-methoxypyridin-3-amine (2.10 g, 13.8 mmol) and potassium thiocyanate (10.73 g, 110.4 mmol) in 30 mL of glacial acetic acid, slowly drop bromine (1.29 g, 20.7 mmol) at 10-15°C, and then stir at room temperature for 2 hours. Add 100 mL of water and 100 mL of ethyl acetate to dilute, filter, separate the filtrate, and wash the organic phase with saturated sodium thiosulfate solution and saturated sodium bicarbonate solution, respectively.

[0288] After concentration, the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 10-1 / 2) to obtain 1.65 g of the title compound as a light yellow solid. The yield was 57.2%.

[0289] LC-MS: m / z = 210.2 [M+H] + .

[0290] Step 4: Preparation of 2-amino-6-ethylthiazolo[5,4-b]pyridin-5(4H)-one (5d).

[0291] 6-Ethyl-5-methoxythiazolo[5,4-b]pyridin-2-amine (1.65 g, 7.89 mmol) was dissolved in 48% hydrobromic acid (15 mL) and stirred at 120°C for 2 hours. The solvent was removed by rotary evaporation, and the residue was dissolved in 30 mL of water, adjusted to pH = 8 with saturated sodium carbonate solution, extracted with ethyl acetate, concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE = 1 / 10 -1 / 2) to obtain 1.28 g of the title compound as a light yellow solid, with a yield of 83.1%.

[0292] LC-MS: m / z = 196.2 [M+H] + .

[0293] Step 5: Preparation of 2-bromo-6-ethylthiazolo[5,4-b]pyridin-5(4H)-one (5e).

[0294] 2-Amino-6-ethylthiazolo[5,4-b]pyridin-5(4H)-one (1.28 g, 4.94 mmol) was dissolved in acetonitrile (20 mL), tert-butyl nitrite (866 mg, 8.40 mmol) and copper bromide (1.88 g, 8.40 mmol) were slowly added in sequence at 0°C, and the mixture was stirred at room temperature for 2 hours after the addition was completed. The reaction was detected to be complete by TLC, and 60 mL of water and 60 mL of ethyl acetate were added to dilute the mixture, filtered, the filtrate was separated, the organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 10-1 / 2) to obtain 1.10 g of the title compound as a light yellow solid, with a yield of 85.9%.

[0295] LC-MS: m / z=261.1[M+2H] + .

[0296] Step 6: Preparation of methyl 6-ethyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carboxylate (5f).

[0297] 2-Bromo-6-ethylthiazolo[5,4-b]pyridin-5(4H)-one (800 mg, 3.09 mmol), triethylamine (469 mg, 4.64 mmol) and bistriphenylphosphine palladium dichloride (109 mg, 0.155 mmol) were dissolved in 5 mL of methanol and 10 mL of acetonitrile, and reacted at 60°C for 16 hours under carbon monoxide atmosphere (0.6 MPa). The reaction was complete by TLC detection, and the reaction solution was concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 10-1 / 2) to obtain 430 mg of the title compound as a yellow solid, with a yield of 58.4%.

[0298] LC-MS: m / z = 239.2 [M+H] + .

[0299] Step 7: Preparation of 6-ethyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carbaldehyde (5 g).

[0300] Methyl 6-ethyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carboxylate (400 mg, 1.68 mmol) was dissolved in dichloromethane (15 mL). Diisobutylaluminum hydride (3.36 mL, 3.36 mmol, 1 Min Hexane) was slowly added dropwise at -78°C. The mixture was stirred for 2 hours at -78°C. The reaction was complete when detected by TLC. An appropriate amount of sodium sulfate decahydrate was slowly added at -78°C to quench the reaction (until no bubbles were emitted). The mixture was stirred for 10 minutes, filtered, and the filtrate was concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 5) to obtain 280 mg of the title compound as a yellow solid in a yield of 80.2%.

[0301] LC-MS: m / z = 209.2 [M+H] + .

[0302] Step 8: Preparation of 6-ethyl-2-(hydroxymethyl)thiazolo[5,4-b]pyridin-5(4H)-one (5h).

[0303] Dissolve 6-ethyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridine-2-carboxaldehyde (280 mg, 1.34 mmol) in anhydrous methanol (10 mL), slowly add sodium borohydride (56.7 mg, 1.5 mmol) at 0°C, and react at room temperature for 0.5 hours. Dilute with 20 mL of water, extract with ethyl acetate, and concentrate the organic phase to obtain 240 mg of the title compound as a white solid, with a yield of 85.4%.

[0304] LC-MS: m / z = 211.2 [M+H] + .

[0305] Step 9: Preparation of 2-(chloromethyl)-6-ethylthiazolo[5,4-b]pyridin-5(4H)-one (5i).

[0306] 6-Ethyl-2-(hydroxymethyl)thiazolo[5,4-b]pyridin-5(4H)-one (240 mg, 1.14 mmol) was dissolved in dichloromethane (10 mL), thionyl chloride (680 mg, 5.71 mmol) and 1 drop of N,N-dimethylformamide were added, and the mixture was reacted at room temperature for 4 hours. The reaction solution was concentrated, and the residue was slurried with n-hexane to obtain 160 mg of the title compound as a brown solid, with a yield of 61.3%.

[0307] LC-MS: m / z = 229.7 [M+H] + .

[0308] Step 10: Preparation of 5-(4-((6-ethyl-5-oxo-4,5-dihydrothiazolo[5,4-b]pyridin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (5).

[0309] At room temperature, 2-(chloromethyl)-6-ethylthiazolo[5,4-b]pyridin-5(4H)-one (160 mg, 0.700 mmol) and N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride (215 mg, 0.840 mmol) were dissolved in acetonitrile (10 mL), N,N-diisopropylethylamine (452 ​​mg, 3.50 mmol) and sodium iodide (10.5 mg, 0.070 mmol) were added, and stirred at 80°C for 4 hours. The reaction solution was concentrated, and the residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid, 30 min), to obtain 5.00 mg of the title compound as a white solid, with a yield of 1.73%.

[0310] LC-MS: m / z=413.5[M+H] + .

[0311] 1 H NMR (400MHz, DMSO-d6) δ8.41(d,J=5.7Hz,1H),8.28(d,J=2.9Hz,1H),7.90(s,1H),7.83(d,J=8.7Hz,1H),7.41(dd,J=8.8,2.9 Hz,1H),3.92(s,2H),3.32(s,4H),2.78(d,J=4.8Hz,3H),2.70(t,J=5.0Hz,4H),2.54(t,J=7.4Hz,2H),1.16(t,J=7.4Hz,3H).

[0312] Example 6: Preparation of 1'-((6-ethyl-5-oxo-4,5-dihydrothieno[3,2-b]pyridin-2-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide (6)

[0313]

[0314] Step 1: Preparation of 1'-(tert-butyl)6-methyl 3',6'-dihydro-[3,4'-bipyridyl]-1',6(2'H)-dicarboxylate (6a).

[0315] 5-Bromopyridine-2-carboxylic acid methyl ester (1.08g, 5.00mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (1.86g, 6.00mmol), [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium (181mg, 0.25mmol) and sodium carbonate (1.59g, 15.0mmol) were dissolved in 16mL of dioxane and 4mL of water, and stirred at 90°C overnight under nitrogen protection. The reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 5) to obtain 820mg of the title compound as a white solid, with a yield of 51.6%.

[0316] LC-MS: m / z=319.4[M+H] + .

[0317] Step 2: Preparation of tert-butyl 6-(methylcarbamoyl)-3',6'-dihydro-[3,4'-bipyridine]-1'(2'H)-carboxylate (6b).

[0318] 1'-(tert-butyl) 6-methyl 3',6'-dihydro-[3,4'-bipyridine]-1',6(2'H)-dicarboxylate (820 mg, 2.57 mmol) was dissolved in 10 mL of 40% aqueous methylamine solution and stirred at room temperature overnight. The reaction solution was concentrated and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE = 1 / 20 -1 / 5) to obtain 760 mg of the title compound as a white solid with a yield of 93.3%.

[0319] LC-MS: m / z=318.4[M+H] + .

[0320] Step 3: Preparation of N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide hydrochloride (6c).

[0321] Dissolve tert-butyl 6-(methylcarbamoyl)-3',6'-dihydro-[3,4'-bipyridine]-1'(2'H)-carboxylate (760 mg, 2.39 mmol) in 10 mL of hydrogen chloride-dioxane solution (4 mol / L), stir overnight at room temperature, and concentrate under reduced pressure to obtain 540 mg of the title compound as a light yellow solid, with a yield of 89.1%.

[0322] LC-MS: m / z = 218.4 [M+H] + .

[0323] Step 4: Preparation of 1'-((6-ethyl-5-oxo-4,5-dihydrothieno[3,2-b]pyridin-2-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide (6).

[0324] At room temperature, 2-(Chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (65.0 mg, 0.287 mmol) and N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide hydrochloride (80.0 mg, 0.315 mmol) were dissolved in acetonitrile (10 mL), N,N-diisopropylethylamine (186 mg, 1.44 mmol) and sodium iodide (4.50 mg, 0.03 mmol) were added, and the mixture was stirred at 80°C for 4 hours. The reaction solution was concentrated and the residue was separated by high pressure preparative liquid separation (chromatographic column model: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid, 30min) to obtain 7.90 mg of the title compound as a white solid, with a yield of 6.75%.

[0325] LC-MS: m / z = 409.5 [M+H] + .

[0326] 1 H NMR (400MHz, DMSO-d6) δ12.04(s,1H),8.78–8.66(m,2H),8.04–7.94(m,2H),7.75(s,1H),6.90(s,1H),6.43(d,J=3.5Hz,1H),3.86(s ,2H),3.21(q,J=3.0Hz,2H),2.81(d,J=4.8Hz,3H),2.73(t,J=5.6Hz,2H),2.55(s,2H),2.44(q,J=7.3Hz,2H),1.12(t,J=7.4Hz,3H).

[0327] Example 7: Preparation of 6-ethyl-2-((4-(1-methyl-1H-indazol-5-yl)piperazin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (7)

[0328]

[0329] Step 1: Preparation of tert-butyl 4-(1-methyl-1H-indazol-5-yl)piperazine-1-carboxylate (7a).

[0330] At room temperature, methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl)palladium(II) (198 mg, 237 μmol) was added to a suspension of 5-bromo-1-methyl-1H-indazole (500 mg, 2.37 mmol) and cesium carbonate (1.54 g, 4.74 mmol) in 1,4-dioxane (10.0 mL). The reaction system was replaced with nitrogen three times and stirred at 110°C for 16 hours under nitrogen atmosphere. The mixture was diluted with water (100 mL) and extracted with ethyl acetate (100 mL x 2). The organic phase was concentrated and the residual compound was separated and purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 0%-40%) to obtain 605 mg of the title compound as a white solid.

[0331] LC-MS: m / z=317[M+H] + .

[0332] Step 2: Preparation of 1-methyl-5-(piperazin-1-yl)-1H-indazole (7b).

[0333] Dissolve tert-butyl 4-(1-methyl-1H-indazol-5-yl)piperazine-1-carboxylate (605 mg, 1.91 mmol) in methanolic hydrogen chloride solution (12.0 mL, 4 M), stir at room temperature for 2 hours, and concentrate under reduced pressure to obtain 610 mg of the title compound as a white solid.

[0334] LC-MS: m / z=217[M+H] + .

[0335] Step 3: Preparation of 6-ethyl-2-((4-(1-methyl-1H-indazol-5-yl)piperazin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (7).

[0336] 1-Methyl-5-(piperazin-1-yl)-1H-indazole (157 mg, 724 μmol) and triethylamine (99 μL, 724 μmol) were dissolved in dichloromethane (5 mL), stirred at room temperature for 15 minutes, glacial acetic acid (207 μL, 3.62 mmol) and 6-ethyl-5-oxo-4,5-dihydrothieno[3,2-b]pyridine-2-carbaldehyde (150 mg, 724 μmol) were added, stirred at room temperature for 2 hours, sodium triacetoxyborohydride (199 mg, 941 μmol) was added, and stirred at room temperature for 16 hours. After concentration under reduced pressure, the residual compound was separated and purified by silica gel column chromatography (mobile phase: DCM: MeOH = 0%-15%), and then separated again by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 14.6 mg of the title compound as a white solid. Yield: 4.95%.

[0337] LC-MS: m / z=408.00[M+H] + .

[0338] 1 H NMR (400MHz, DMSO-d6) δ12.03(s,1H),7.85(s,1H),7.75(s,1H),7.49(d,J=9.2Hz,1H),7.23(dd,J=9.2,2.0Hz,1H),7.06(d,J=2.0Hz ,1H),6.88(s,1H),3.97(s,3H),3.79(s,2H),3.17–3.00(m,4H),2.62(t,J=4.8Hz,4H),2.44(q,J=7.6Hz,2H),1.12(t,J=7.6Hz,3H).

[0339] Example 8: Preparation of 6-ethyl-2-((4-(imidazo[1,2-a]pyridin-6-yl)piperazin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (8)

[0340]

[0341] Step 1: Preparation of tert-butyl 4-(imidazo[1,2-a]pyridin-6-yl)piperazine-1-carboxylate (8a).

[0342] 6-Bromoimidazo[1,2-a]pyridine (590 mg, 3.00 mmol), N-Boc-piperazine (607 mg, 3.60 mmol), Pd2(dba)3 (137 mg, 0.15 mmol), Xantphos (173 mg, 0.30 mmol) and cesium carbonate (3.17 g, 9.00 mmol) were dissolved in 16 mL of dioxane and 4 mL of water, and stirred at 90°C overnight under nitrogen protection. The reaction solution was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE = 1 / 20 -1 / 5) to obtain 720 mg of the title compound as a white solid, with a yield of 80.0%.

[0343] LC-MS: m / z=303.2[M+H] + .

[0344] Step 2: Preparation of 6-(piperazin-1-yl)imidazo[1,2-a]pyridine (8b).

[0345] Dissolve tert-butyl 4-(imidazo[1,2-a]pyridin-6-yl)piperazine-1-carboxylate (720 mg, 2.41 mmol) in 10 mL of dichloromethane, add 3 mL of trifluoroacetic acid, and stir at room temperature for 2 hours. Concentrate under reduced pressure, dissolve the residue in 20 mL of dichloromethane, wash with saturated sodium carbonate aqueous solution, and concentrate to obtain 380 mg of the title compound as a brown oil, with a yield of 79.1%.

[0346] LC-MS: m / z = 203.2 [M+H] + .

[0347] Step 3: Preparation of 6-ethyl-2-((4-(imidazo[1,2-a]pyridin-6-yl)piperazin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (8).

[0348] At room temperature, 2-(chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (114 mg, 0.502 mmol) and 6-(piperazin-1-yl)imidazo[1,2-a]pyridine (120 mg, 0.602 mmol) were dissolved in acetonitrile (10 mL), N,N-diisopropylethylamine (324 mg, 2.51 mmol) and sodium iodide (7.50 mg, 0.050 mmol) were added, and the mixture was stirred at 80°C for 4 hours. The mixture was concentrated under reduced pressure, and the residue was separated by high pressure preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid, 30 min) to obtain 11.00 mg of the title compound as a white solid, with a yield of 5.60%.

[0349] LC-MS: m / z=394.1[M+H] + .

[0350] 1 H NMR (400MHz, DMSO-d6) δ12.25(s,1H),8.95(s,1H),8.23(s,1H),8.17–7.99(m,2H),7.97–7.81(m,2H),7.21(s ,1H),6.44(s,1H),4.75–4.50(m,2H),3.82(s,2H),2.78(s,2H),2.46(d,J=7.4Hz,4H),1.14(t,J=7.4Hz,3H).

[0351] Example 9: Preparation of 5-((1-((6-ethyl-5-oxo-4,5-dihydrothieno[3,2-b]pyridin-2-yl)methyl)azetidin-3-yl)oxy)-N-methylpicolinamide (9)

[0352]

[0353] Step 1: Preparation of methyl 5-((1-benzhydrylazetidin-3-yl)oxy)picolinate (9a).

[0354] Methyl 5-fluoropicolinate (500 mg, 3.22 mmol), 1-benzhydryl-3-hydroxyazetidine (1.54 g, 6.45 mmol) and cesium carbonate (2.10 g, 6.45 mmol) were dissolved in acetonitrile (20 mL) and stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure and the residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate: petroleum ether = 40%-70%) to obtain 2.60 g of the title compound as a yellow solid.

[0355] LC-MS: m / z=375.5[M+H] + .

[0356] Step 2: Preparation of 5-((1-benzhydrylazetidin-3-yl)oxy)-N-methylpicolinamide (9b).

[0357] Dissolve methyl 5-((1-benzhydrylazetidin-3-yl)oxy)picolinate (2.50 g, 6.68 mmol) in methanol (25 mL), add 40% methylamine aqueous solution (6.0 mL, 68.8 mmol), and stir at room temperature for 2 hours. Concentrate under reduced pressure, and the residual compound is separated and purified by silica gel column chromatography (mobile phase: dichloromethane:methanol=0%-15%) to obtain 3.40 g of the title compound as a yellow solid.

[0358] LC-MS: m / z=374.5[M+H] + .

[0359] Step 3: Preparation of 5-(azetidin-3-yloxy)-N-methylpicolinamide (9c).

[0360] Dissolve 5-((1-benzhydrylazetidin-3-yl)oxy)-N-methylpicolinamide (3.30 g, 8.84 mmol) and trifluoroacetic acid (1.1 mL) in methanol (33 mL), add palladium hydroxide (3.00 g) and cesium carbonate (2.10 g, 6.45 mmol), replace the hydrogen three times, and stir at room temperature under hydrogen atmosphere (15 Psi) for 2 hours. Filter, concentrate the filtrate, and separate and purify the residue by silica gel column chromatography (mobile phase: dichloromethane: methanol = 5%-15%) to obtain 2.21 g of the title compound as a colorless oil.

[0361] LC-MS: m / z = 208.3 [M+H] + .

[0362] Step 4: Preparation of 5-((1-((6-ethyl-5-oxo-4,5-dihydrothieno[3,2-b]pyridin-2-yl)methyl)azetidin-3-yl)oxy)-N-methylpicolinamide (9).

[0363] N,N-diisopropylethylamine (77 μL, 439 μmol) and sodium iodide (65.8 mg, 439 μmol) were added to a solution of 2-(chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (50.0 mg, 220 μmol) and 5-(azetidin-3-yloxy)-N-methylpicolinamide (50.1 mg, 242 μmol) in acetonitrile (1.0 mL), and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: DCM:MeOH=5%-15%), and then separated again by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 6.7 mg of the title compound as a white solid. Yield: 7.66%.

[0364] LC-MS: m / z=399.00[M+H] + .

[0365] 1H NMR (400MHz, DMSO-d6) δ12.23(s,1H),8.58(q,J=4.8Hz,1H),8.26(d,J=2.8Hz,1H),7.99(d,J=8.8Hz,1H),7.86(s,1H),7.46(dd,J=8.8,2.8H z,1H),7.21(s,1H),5.22(s,1H),4.71(s,2H),4.58(s,2H),4.23(s,2H),2.80(d,J=4.8Hz,3H),2.45(t,J=7.2Hz,2H),1.12(t,J=7.2Hz,3H).

[0366] Example 10: Preparation of 6-ethyl-2-((3-((1-methyl-1H-indazol-5-yl)oxy)azetidin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (10)

[0367]

[0368]

[0369] Step 1: Preparation of 1-methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (10a).

[0370] 5-Bromo-1-methyl-1H-indazole (1.05 g, 5.00 mmol), biboronic acid pinacol ester (1.52 g, 6.00 mmol), [1,1'-bis(diphenylphosphino)ferrocene] palladium dichloride (181 mg, 0.25 mmol) and potassium acetate (1.47 g, 15.0 mmol) were dissolved in 20 mL of dioxane and stirred at 90°C for 4 hours under nitrogen protection. The mixture was concentrated under reduced pressure and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE = 1 / 20 -1 / 5) to obtain 1.16 g of the title compound as a white solid with a yield of 89.9%.

[0371] LC-MS: m / z = 259.1 [M+H] + .

[0372] Step 2: Preparation of 1-methyl-1H-indazol-5-ol (10b).

[0373] 1-Methyl-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (1.16 g, 4.49 mmol) was dissolved in methanol (10 mL), 30% hydrogen peroxide (2.55 g, 22.5 mmol) was slowly added dropwise at 0°C, and stirred at room temperature for 4 hours. 30 mL of water was added for dilution, and ethyl acetate was used for extraction. The organic phase was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE = 1 / 10 -1 / 2) to obtain 0.64 g of the title compound as a light yellow solid, with a yield of 96.2%.

[0374] LC-MS: m / z = 149.2 [M+H] + .

[0375] Step 3: Preparation of tert-butyl 3-((1-methyl-1H-indazol-5-yl)oxy)azetidine-1-carboxylate (10c).

[0376] 1-Methyl-1H-indazol-5-ol (300 mg, 2.02 mmol), cesium carbonate (1.65 g, 5.05 mmol) and tert-butyl 3-iodoazetidine-1-carboxylate (685 mg, 2.42 mmol) were dissolved in 20 mL of acetonitrile and stirred at 80° C. for 2 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 10-1 / 2) to obtain 360 mg of the title compound as a light yellow solid, with a yield of 58.7%.

[0377] LC-MS: m / z=304.4[M+H] + .

[0378] Step 4: Preparation of 5-(azetidin-3-yloxy)-1-methyl-1H-indazole hydrochloride (10d).

[0379] Tert-butyl 3-((1-methyl-1H-indazol-5-yl)oxy)azetidine-1-carboxylate (360 mg, 1.19 mmol) was dissolved in 10 mL of hydrogen chloride-dioxane solution (4 mol / L), stirred at room temperature overnight, and concentrated under reduced pressure to obtain 280 mg of the title compound as a light yellow solid, with a yield of 98.2%.

[0380] LC-MS: m / z = 204.4 [M+H] + .

[0381] Step 5: Preparation of 6-ethyl-2-((3-((1-methyl-1H-indazol-5-yl)oxy)azetidin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (10).

[0382] 2-(Chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (111 mg, 0.487 mmol), 5-(azetidin-3-yloxy)-1-methyl-1H-indazole hydrochloride (140 mg, 0.584 mmol), N,N-diisopropylethylamine (315 mg, 2.44 mmol) and sodium iodide (7.50 mg, 0.050 mmol) were dissolved in acetonitrile (10 mL) and stirred at 80°C for 4 hours. The mixture was concentrated under reduced pressure and the residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid, 30 min) to obtain 8.00 mg of the title compound as a white solid with a yield of 4.16%.

[0383] LC-MS: m / z=395.5[M+H] + .

[0384] 1 H NMR (400MHz, DMSO-d6) δ11.99(s,1H),7.86(s,1H),7.74(s,1H),7.55(d,J=8.8Hz,1H),7.05–6.97(m,2H),6.84(s,1H),4.84(p,J=5.6H z,1H),3.99(s,3H),3.88(s,2H),3.82(dd,J=8.2,5.9Hz,2H),3.13(dd,J=7.7,5.8Hz,2H),2.44(q,J=7.4Hz,2H),1.11(t,J=7.4Hz,3H).

[0385] Example 11: Preparation of 5-((1-((6-ethyl-5-oxo-4,5-dihydrothieno[3,2-b]pyridin-2-yl)methyl)pyrrol-3-yl)oxy)-N-methylpicolinamide (11)

[0386]

[0387] Step 1: Preparation of methyl 5-((1-(tert-butoxycarbonyl)pyrrol-3-yl)oxy)picolinate (11a).

[0388] Methyl 5-fluoropicolinate (500 mg, 3.22 mmol), N-tert-butyloxycarbonyl-3-pyrrolidinol (1.81 g, 9.678 mmol) and cesium carbonate (2.10 g, 6.45 mmol) were dissolved in acetonitrile and stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure and the residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether: ethyl acetate = 40%-70%) to obtain 1.10 g of the title compound as a yellow solid.

[0389] LC-MS: m / z=323.4[M+H] + .

[0390] Step 2: Preparation of tert-butyl 3-((6-(methylcarbamoyl)pyridin-3-yl)oxy)pyrrole-1-carboxylate (11b).

[0391] Dissolve 5-((1-(tert-butyloxycarbonyl)pyrrol-3-yl)oxy)picolinic acid methyl ester (1.00 g, 3.10 mmol) in methanol (10 mL), add 40% methylamine aqueous solution (2.79 mL, 32.0 mmol), and stir at room temperature for 2 hours. Concentrate under reduced pressure, and the residue is separated and purified by silica gel column chromatography (mobile phase: dichloromethane: methanol)

[0392] =0%-15%) to give 1.02 g of the title compound as a white solid.

[0393] LC-MS: m / z=322.3[M+H] + .

[0394] Step 3: Preparation of 5-(pyrrol-3-yloxy)-N-methylpicolinamide (11c).

[0395] Dissolve tert-butyl 3-((6-(methylcarbamoyl)pyridin-3-yl)oxy)pyrrole-1-carboxylate (1.00 g, 3.10 mmol) in a solution of hydrogen chloride in dioxane (9.0 mL, 4 M) and stir at room temperature for 2 hours. Concentrate under reduced pressure to obtain 891 mg of the title compound as a white solid. Yield: 4.95%.

[0396] LC-MS: m / z = 222.3 [M+H] + .

[0397] Step 4: Preparation of 5-((1-((6-ethyl-5-oxo-4,5-dihydrothieno[3,2-b]pyridin-2-yl)methyl)pyrrol-3-yl)oxy)-N-methylpicolinamide (11).

[0398] 2-(Chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (50.0 mg, 220 μmol), 5-(pyrrol-3-yloxy)-N-methylpicolinamide (57.0 mg, 220 μmol), N,N-diisopropylethylamine (191 μL, 1.10 mmol) and sodium iodide (66.0 mg, 439 μmol) were dissolved in acetonitrile (5.0 mL) and stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residual compound was separated and purified by silica gel column chromatography (mobile phase: DCM:MeOH=0%-15%), and then separated again by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 25.8 mg of the title compound as a white solid. Yield: 28.5%.

[0399] LC-MS: m / z 413.10 [M+H] + .

[0400] 1 H NMR (400MHz, DMSO-d6) δ12.05(s,1H),8.56(q,J=4.8Hz,1H),8.24(d,J=2.8Hz,1H),7.9 4(d,J=8.8Hz,1H),7.73(s,1H),7.47(dd,J=8.8,2.8Hz,1H),6.85(s,1H),5.06(dq,J=6 .8,2.8Hz,1H),3.87(s,2H),2.94(dd,J=10.8,6.0Hz,1H),2.86–2.71(m,5H),2.52(s,1 H),2.43(q,J=7.2Hz,2H),2.38–2.30(m,1H),1.89–1.76(m,1H),1.11(t,J=7.2Hz,3H).

[0401] Example 12: Preparation of 6-ethyl-2-((4-(1-methyl-1H-indazol-5-yl)-3,6-dihydropyridin-1(2H)-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (12)

[0402]

[0403] Step 1: Preparation of tert-butyl 4-(1-methyl-1H-indazol-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (12a).

[0404] 5-Bromo-1-methyl-1H-indazole (0.63 g, 2.98 mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (1.11 g, 3.58 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (109 mg, 0.15 mmol) and sodium carbonate (948 mg, 8.94 mmol) were dissolved in 16 mL of dioxane and 4 mL of water, and stirred at 90° C. overnight under nitrogen protection. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 5) to obtain 560 mg of the title compound as a white solid, with a yield of 60.0%.

[0405] LC-MS: m / z=314.4[M+H] + .

[0406] Step 2: Preparation of 1-methyl-5-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole hydrochloride (12b).

[0407] Dissolve tert-butyl 4-(1-methyl-1H-indazol-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (560 mg, 1.79 mmol) in 10 mL of hydrogen chloride-dioxane solution (4 mol / L), stir overnight at room temperature, and concentrate under reduced pressure to obtain 430 mg of the title compound as a light yellow solid, with a yield of 96.2%.

[0408] LC-MS: m / z = 214.3 [M+H] + .

[0409] Step 3: Preparation of 6-ethyl-2-((4-(1-methyl-1H-indazol-5-yl)-3,6-dihydropyridin-1(2H)-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (12).

[0410] 2-(Chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (66.3 mg, 0.291 mmol), 1-methyl-5-(1,2,3,6-tetrahydropyridin-4-yl)-1H-indazole hydrochloride (80.0 mg, 0.320 mmol), N,N-diisopropylethylamine (188 mg, 1.46 mmol) and sodium iodide (4.35 mg, 0.029 mmol) were dissolved in acetonitrile (10 mL) and stirred at 80°C for 4 hours. The mixture was concentrated under reduced pressure and the residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid, 30 min) to obtain 16.0 mg of the title compound as a white solid with a yield of 13.6%.

[0411] LC-MS: m / z=405.5[M+H] + .

[0412] 1 H NMR (400MHz, DMSO-d6) δ12.27(s,1H),8.06(s,1H),7.90(s,1H),7.81(s,1H),7.68–7.57(m,2H),7.27(s,1H),6.20(d,J=3.7Hz, 1H),4.74(s,2H),4.04(s,3H),3.87(s,2H),3.71(s,1H),3.30(s,1H),2.88(s,2H),2.46(d,J=7.4Hz,2H),1.14(t,J=7.4Hz,3H).

[0413] Example 13: Preparation of 6-ethyl-2-((3-((1-methyl-1H-indazol-5-yl)oxy)pyrrol-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (13)

[0414]

[0415]

[0416] Step 1: Preparation of 6-ethyl-2-((3-((1-methyl-1H-indazol-5-yl)oxy)pyrrol-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (13).

[0417] 2-(Chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (40.0 mg, 176 μmol), 1-

[0418] Methyl-5-(pyrrol-3-yloxy)-1H-indazole (70.5 mg, 211 μmol), N,N-diisopropylethylamine (61 μL, 351 μmol) and sodium iodide (5.30 mg, 35.1 μmol) were dissolved in acetonitrile (5 mL) and stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 26.5 mg of the title compound as a white solid. Yield: 46.9%.

[0419] LC-MS: m / z=409.15[M+H] + .

[0420] 1 H NMR (400MHz, DMSO-d6) δ10.75(s,1H),7.92(s,1H),7.87(s,1H),7.58(d,J=9.2Hz,1H),7.24(s,1H),7.23(d,J=2.4Hz,1H),7.06(dd,J=9.2,2.4Hz ,1H),5.16(s,1H),4.73(s,2H),4.01(s,3H),3.66–3.58(m,2H),2.53(d, J=6.8Hz,2H),2.46(t,J=7.6Hz,2H),2.20(m,2H),1.13(t,J=7.6Hz,3H).

[0421] Example 14: Preparation of 6-ethyl-2-((4-(imidazo[1,2-a]pyridin-6-yl)-3,6-dihydropyridin-1(2H)-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (14)

[0422]

[0423] Step 1: Preparation of tert-butyl 4-(imidazo[1,2-a]pyridin-6-yl)-3,6-dihydropyridine-1(2H)-carboxylate (14a).

[0424] 6-Bromoimidazo[1,2-a]pyridine (590 mg, 3.00 mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (1.11 g, 3.60 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (109 mg, 0.15 mmol) and sodium carbonate (954 mg, 9.00 mmol) were dissolved in 16 mL of dioxane and 4 mL of water, and stirred at 90° C. overnight under nitrogen protection. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase EA / PE=1 / 20-1 / 5) to obtain 720 mg of the title compound as a white solid, with a yield of 80.2%.

[0425] LC-MS: m / z=300.4[M+H] + .

[0426] Step 2: Preparation of 6-(1,2,3,6-tetrahydropyridin-4-yl)imidazo[1,2-a]pyridine (14b).

[0427] Tert-butyl 4-(1-methyl-1H-indazol-5-yl)-3,6-dihydropyridine-1(2H)-carboxylate (720 mg, 2.41 mmol) was dissolved in 10 mL of dichloromethane, 3 mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residue was dissolved in 20 mL of dichloromethane, washed with saturated aqueous sodium carbonate solution, and concentrated to obtain 380 mg of the title compound as a brown oil, with a yield of 79.2%.

[0428] LC-MS: m / z = 200.4 [M+H] + .

[0429] Step 3: Preparation of 6-ethyl-2-((4-(imidazo[1,2-a]pyridin-6-yl)-3,6-dihydropyridin-1(2H)-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (14).

[0430] 2-(Chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (114 mg, 0.502 mmol), N,N-diisopropylethylamine (324 mg, 2.51 mmol), sodium iodide (7.50 mg, 0.050 mmol) and 6-(1,2,3,6-tetrahydropyridin-4-yl)imidazo[1,2-a]pyridine (120 mg, 0.602 mmol) were dissolved in acetonitrile (10 mL) and stirred at 80°C for 4 hours. The mixture was concentrated under reduced pressure and the residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid, 30 min) to obtain 9.00 mg of the title compound as a white solid with a yield of 4.59%.

[0431] LC-MS: m / z=391.5[M+H] + .

[0432] 1 H NMR(400MHz,DMSO-d6)δ12.25(s,1H),8.95(s,1H),8.23(s,1H),8.17–

[0433] 7.99(m,2H),7.97–7.81(m,2H),7.21(s,1H),6.44(s,1H),4.75–4.50(m,2 H), 3.82 (s, 2H), 2.78 (s, 2H), 2.46 (d, J = 7.4Hz, 4H), 1.14 (t, J = 7.4Hz, 3H).

[0434] Example 15: Preparation of 6-ethyl-2-((3-(imidazo[1,2-a]pyridin-6-yloxy)azetidin-1-yl)methyl)thiophene[3,2-b]pyridin-5(4H)-one (15)

[0435]

[0436] Step 1: Preparation of tert-butyl 3-((6-nitropyridin-3-yl)oxy)azetidine-1-carboxylate (15a).

[0437] 5-Fluoro-2-nitropyridine (284 mg, 2.00 mmol), tert-butyl 3-hydroxyazetidine-1-carboxylate (415 mg, 2.40 mmol) and cesium carbonate (975 mg, 3.00 mmol) were dissolved in tetrahydrofuran (5 mL) and reacted at room temperature for 3 hours, and then at 50°C for 20 hours. The mixture was filtered, the filtrate was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 5%-10%-20%) to obtain 654 mg of the title compound as a light yellow solid with a yield of >99%.

[0438] Step 2: Preparation of tert-butyl 3-((6-aminopyridin-3-yl)oxy)azetidine-1-carboxylate (15b).

[0439] tert-Butyl 3-((6-nitropyridin-3-yl)oxy)azetidine-1-carboxylate (650 mg, 2.18 mmol), reduced iron powder (617 mg, 10.9 mmol) and glacial acetic acid (10 mL) were added to a reaction bottle, stirred at 80°C for 3 hours, concentrated under reduced pressure, methanol (10 mL) and dichloromethane (10 mL) were added to the residue, stirred at room temperature for 0.5 hour, filtered, and the filtrate was concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane: 2%-3%-5%) to obtain 360 mg of the title compound as a light brown solid with a yield of 61.6%.

[0440] LC-MS: m / z = 266.0 [M+H] + .

[0441] Step 3: Preparation of tert-butyl 3-(imidazo[1,2-a]pyridin-6-yloxy)azetidine-1-carboxylate (15c).

[0442] Dissolve tert-butyl 3-((6-aminopyridin-3-yl)oxy)azetidine-1-carboxylate (360 mg, 1.36 mmol) and 2-chloroacetaldehyde (160 mg, 7.03 mmol) in anhydrous ethanol (5 mL) and water (1 mL), stir at 80 ° C for 16 hours under nitrogen protection, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane: 2%-5%-10%) to give the title compound as a light brown solid, 160 mg, yield 40.8%.

[0443] LC-MS: m / z = 290.0 [M+H] + .

[0444] Step 4: Preparation of 6-(azetidin-3-yloxy)imidazo[1,2-a]pyridine hydrochloride (15d).

[0445] Dissolve tert-butyl 3-(imidazo[1,2-a]pyridin-6-yloxy)azetidine-1-carboxylate (160 mg, 0.554 mmol) in a solution of hydrogen chloride in dioxane (10 mL, 4 mol / L), stir at room temperature for 12 hours, and concentrate under reduced pressure. The residue is washed with methanol three times (10 mL×3) to give 160 mg of the title compound as a light yellow solid.

[0446] Step 5: Preparation of 6-ethyl-2-((3-(imidazo[1,2-a]pyridin-6-yloxy)azetidin-1-yl)methyl)thiophen[3,2-b]pyridin-5(4H)-one (15).

[0447] 2-(Chloromethyl)-6-ethylthieno[3,2-b]pyridin-5(4H)-one (50.0 mg, 0.220 mmol), 6-(azetidin-3-yloxy)imidazo[1,2-a]pyridine hydrochloride (60 mg, 0.267 mmol), sodium iodide (6.4 mg, 0.0450 mmol) and diisopropylethylamine (0.4 mL, 2.25 mmol) were dissolved in acetonitrile (5 mL), stirred at room temperature for 16 hours, concentrated under reduced pressure, and the residue was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um, 100 A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain the title compound as a brown-red solid, 44.0 mg, yield 52.6%.

[0448] LC-MS: m / z=381.0[M+H] + .

[0449] 1H-NMR (400MHz, d6-DMSO) δ8.54(d,J=2.0Hz,1H),8.15(s,2H),7.94(d,J=9.6Hz,1H),7.86(s,1H),7.70(dd,J1=10.0Hz,J2=2. 0Hz,1H),7.21(s,1H),5.11-5.13(m,1H),4.55-4.59(m,2H),4.23-4.28(m,2H),2.46(q,J=7.2Hz,2H),1.13(t,J=7.2Hz,3H).

[0450] Example 16: Preparation of 6-ethyl-2-(3-imidazo[1,2-a]pyridin-6-yloxy)pyrrolidin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (16)

[0451]

[0452] Step 1: Preparation of 6-ethyl-2-(3-imidazo[1,2-a]pyridin-6-yloxy)pyrrolidin-1-yl)methyl)thieno[3,2-b]pyridin-5(4H)-one (16).

[0453] 6-(Pyrrolidin-3-yloxy)imidazo[1,2-a]pyridine hydrochloride (63.3 mg, 0.264 mmol), 2-chloromethyl-6-ethylthieno[3,2-b]pyridin-5(4H)one (50.0 mg, 0.220 mmol), N,N-diisopropylethylamine (171 mg, 1.32 mmol) and sodium iodide (66.0 mg, 0.440 mmol) were dissolved in acetonitrile (5.0 mL) and stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure and the residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid) to obtain 57.0 mg of the title compound as a white solid with a yield of 65.6%.

[0454] LC-MS: m / z=395.1[M+H] + .

[0455] 1H NMR (400MHz, DMSO-d6) δ12.29(s,1H), δ8.68(d,J=2.3Hz,1H),8.16(d,J=8.5Hz,1H),7.92(d,J=9.8Hz,1H),7.86(s,1H),7 .66(dd,J=9.8,2.3Hz,1H),7.22(s,1H),5.17(s,1H),4.71(s,2H),3.58(s,6H),2.48–2.43(m,2H),1.13(t,J=7.4Hz,3H).

[0456] Example 17: Preparation of N-methyl-5-(4-((5-oxo-5,6,7,8-tetrahydro-4H-cyclopentyl[d]thieno[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (17).

[0457]

[0458]

[0459] Step 1: Preparation of methyl 2-((trifluoromethylsulfonyl)oxy)cyclopentyl-1-ene-1-carboxylate (17a).

[0460] Dissolve methyl 2-oxocyclopentane-1-carboxylate (1.00 g, 7.04 mmol) in tetrahydrofuran (30 mL), add sodium hydride (324 mg, 8.10 mmol), stir at 0°C for 30 minutes, add N-phenylbis(trifluoromethanesulfonyl)imide (3.02 g, 8.45 mmol), and stir at room temperature for 16 hours. Quench the reaction with saturated sodium bicarbonate solution (50 mL), extract with ethyl acetate (50 mL x 3), wash the organic phase with saturated brine (50 mL), dry over anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-11%) to obtain 1.80 g of the title compound as a colorless liquid with a yield of 93.1%.

[0461] LC-MS: m / z = 274.9 [M+H] + .

[0462] Step 2: Preparation of methyl 2-(4,4,5,5-tetramethyl-1,3,2-dioxaboronan-2-yl)cyclopentyl-1-ene-1-carboxylate (17b).

[0463] At room temperature and under nitrogen atmosphere, methyl 2-((trifluoromethylsulfonyl)oxy)cyclopentyl-1-ene-1-carboxylate (1.54 g, 5.07 mmol) was dissolved in 1,4-dioxane (12.0 mL), potassium acetate (1.15 g, 11.7 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (128 mg, 0.175 mmol) and biboronic acid pinacol ester (1.63 g, 6.43 mmol) were added, and the mixture was reacted at 80°C for 16 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-5%) to obtain 1.35 g of the title compound as a white solid with a yield of 91.6%.

[0464] LC-MS: m / z = 253.0 [M+H] + .

[0465] Step 3: Preparation of methyl 5-(2-(methoxycarbonyl)cyclopentyl-1-en-1-yl)-4-nitrothiophene-2-carboxylate (17c).

[0466] At room temperature and under nitrogen atmosphere, methyl 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclopentyl-1-ene-1-carboxylate (693 mg, 2.75 mmol) was dissolved in 1,4-dioxane (10 mL) and water (2.5 mL), and methyl 5-bromo-4-nitrothiophene-2-carboxylate (665 mg, 2.50 mmol), potassium carbonate (691 mg, 5.00 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (91.5 mg, 0.125 mmol) were added, and the mixture was reacted at 80°C for 16 hours. Water (15 mL) was added to dilute the mixture, and the mixture was extracted with ethyl acetate (20 mL x 3). The organic phase was washed with saturated brine (20 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-10%) to obtain 526 mg of the title compound as a yellow liquid. The yield was 61.5%.

[0467] LC-MS: m / z 311.9 [M+H] + .

[0468] Step 4: Preparation of methyl 5-oxo-5,6,7,8-tetrahydro-4H-cyclopentyl[d]thieno[3,2-b]pyridine-2-carboxylate (17d).

[0469] Dissolve 5-(2-(methoxycarbonyl)cyclopentyl-1-en-1-yl)-4-nitrothiophene-2-carboxylic acid methyl ester (473 mg, 1.52 mmol) in acetic acid (5 mL), add iron powder (340 mg, 6.08 mmol), and react at 80°C for 2 hours. Concentrate under reduced pressure, add methanol (20 mL), stir for 1 hour, filter, wash the filter cake with methanol (50 mL), concentrate the filtrate, and separate and purify the residue by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-6%) to obtain 214 mg of the title compound as a yellow solid, with a yield of 56.5%.

[0470] LC-MS: m / z 249.9[M+H] + .

[0471] Step 5: Preparation of 2-hydroxymethyl-4,6,7,8-tetrahydro-5H-cyclopentyl[d]thieno[3,2-b]pyridin-5-one (17e).

[0472] At 0°C, under nitrogen atmosphere, methyl 5-oxo-5,6,7,8-tetrahydro-4H-cyclopentyl[d]thieno[3,2-b]pyridine-2-carboxylate (194 mg, 0.778 mmol) was dissolved in tetrahydrofuran (4.0 mL), and a 1.0 M solution of lithium aluminum hydride in tetrahydrofuran (1.6 mL, 1.56 mmol) was added dropwise, and the mixture was reacted at room temperature for 2 hours. Sodium sulfate decahydrate was added to quench the reaction, and the mixture was filtered. The filter cake was washed with methanol (20 mL), and the filtrate was concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-10%) to obtain 148 mg of the title compound as a yellow solid, with a yield of 86.1%.

[0473] LC-MS: m / z 221.9 [M+H] + .

[0474] Step 6: Preparation of 2-chloromethyl-4,6,7,8-tetrahydro-5H-cyclopentyl[d]thieno[3,2-b]pyridin-5-one (17f).

[0475] At room temperature and nitrogen atmosphere, 2-hydroxymethyl-4,6,7,8-tetrahydro-5H-cyclopentyl[d]thieno[3,2-b]pyridin-5-one (128.3 mg, 0.580 mmol) was dissolved in dichloromethane (6 mL), thionyl chloride (690 mg, 5.80 mmol) was added, and a drop of N,N-dimethylformamide was added dropwise, and the mixture was reacted at room temperature for 3 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-5%) to obtain 98.7 mg of the title compound as a yellow solid, with a yield of 61.5%.

[0476] LC-MS: m / z 239.9[M+H] + .

[0477] Step 7: Preparation of N-methyl-5-(4-((5-oxo-5,6,7,8-tetrahydro-4H-cyclopentyl[d]thieno[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (17).

[0478] At room temperature and under nitrogen atmosphere, 2-chloromethyl-4,6,7,8-tetrahydro-5H-cyclopentyl[d]thieno[3,2-b]pyridin-5-one (98.7 mg, 0.412 mmol) and N-methyl-5-piperazin-1-ylpicolinamide hydrochloride (116 mg, 0.453 mmol) were dissolved in acetonitrile (5 mL), and N,N-diisopropylethylamine (320 mg, 2.47 mmol) and sodium iodide (124 mg, 0.824 mmol) were added, and the mixture was reacted at 50°C for 16 hours. After concentration under reduced pressure, the residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid), to obtain 19.0 mg of the title compound as a yellow solid, with a yield of 10.9%.

[0479] LC-MS: m / z = 423.9 [M+H] + .

[0480] 1 H NMR (400MHz, Methanol-d4) δ8.46 (s, 1H), 8.23 ​​(d, J = 9.0Hz, 1H), 7.98 (dd, J = 9.1, 2.5Hz, 1H), 7.59 (s, 1H), 4. 83(s,2H),4.20(s,2H),3.53(m,6H),3.12(m,2H),2.98(s,3H),2.93(t,J=7.5Hz,2H),2.27(p,J=7.6Hz,2H).

[0481] Example 18: Preparation of N-methyl-5-((5-oxo-4,5-dihydrofuro[2,3-d]thieno[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (18).

[0482]

[0483] Step 1: Preparation of methyl 4-amino-5-bromothiophene-2-carboxylate (18a).

[0484] Dissolve 5-bromo-4-nitrothiophene-2-carboxylic acid methyl ester (5.00 g, 18.8 mmol) in 95% ethanol (60 mL), add ammonium chloride (9.05 g, 169 mmol) and iron powder (5.25 g, 94.0 mmol), and stir at 80°C for 2 hours. Filter, wash the filter cake with methanol (100 mL), concentrate the filtrate, and separate and purify the residue by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-10%) to obtain 2.75 g of the title compound as a yellow solid, with a yield of 61.9%.

[0485] LC-MS: m / z=235.8,237.8[M+H] + .

[0486] Step 2: Preparation of methyl 5-bromo-4-(furan-3-carboxamido)thiophene-2-carboxylate (18b).

[0487] At room temperature and under nitrogen atmosphere, 3-furancarboxylic acid (1.69 g, 15.1 mmol) was dissolved in N,N-dimethylformamide (25 mL), and N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)urea hexafluorophosphate (7.19 g, 18.9 mmol) and N,N-diisopropylethylamine (4.07 g, 31.5 mmol) were added. After stirring at room temperature for 10 minutes, 4-amino-5-bromothiophene-2-carboxylic acid methyl ester (2.97 g, 12.6 mmol) was added, and the mixture was stirred at 80°C for 16 hours. Water (75 mL) was added to dilute, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-15%) to obtain 2.77 g of the title compound as a yellow solid. The yield was 66.6%.

[0488] LC-MS: m / z=329.7,331.7[M+H] + .

[0489] Step 3: Preparation of methyl 5-oxo-4,5-dihydrofuro[2,3-d]thieno[3,2-b]pyridine-2-carboxylate (18c).

[0490] At room temperature and under nitrogen atmosphere, methyl 5-bromo-4-(furan-3-carboxamido)thiophene-2-carboxylate (2.72 g, 8.24 mmol) was dissolved in N,N-dimethylacetamide (40.0 mL), potassium acetate (1.62 g, 16.5 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (301 mg, 0.411 mmol) were added, and the mixture was reacted at 100°C for 16 hours. Water (100 mL) was added for dilution, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated sodium chloride solution (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-5%) to obtain 331 mg of the title compound as a yellow solid with a yield of 16.1%.

[0491] LC-MS: m / z = 249.9 [M+H] + .

[0492] Step 4: Preparation of 2-hydroxymethylfuro[2,3-d]thieno[3,2-b]pyridin-5(4H)one (18d).

[0493] At 0°C, under nitrogen atmosphere, methyl 5-oxo-4,5-dihydrofurano[2,3-d]thieno[3,2-b]pyridine-2-carboxylate (100 mg, 0.401 mmol) was dissolved in tetrahydrofuran (5.0 mL), and a 1.0 M solution of lithium aluminum hydride in tetrahydrofuran (1.6 mL, 1.60 mmol) was added dropwise, and the mixture was reacted at room temperature for 2 hours. Sodium sulfate decahydrate (500 mg) was added to quench the reaction, and the mixture was filtered. The filter cake was washed with methanol (30 mL), and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-5%) to obtain 19.0 mg of the title compound as a yellow solid, with a yield of 21.4%. LC-MS: m / z = 221.9 [M+H] + .

[0494] Step 5: Preparation of 5-oxo-4,5-dihydrofuro[2,3-d]thieno[3,2-b]pyridine-2-carbaldehyde (18e).

[0495] At 0°C under nitrogen atmosphere, 2-hydroxymethylfuro[2,3-d]thieno[3,2-b]pyridin-5(4H)one was added.

[0496] (19.0 mg, 0.086 mmol) was dissolved in dichloromethane (10 mL), and Dess-Martin periodinane (73.0 mg, 0.172 mmol) was added, and the mixture was reacted at 40°C for 16 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-5%) to obtain 13.6 mg of the crude title compound as a yellow solid, with a yield of 72.1%.

[0497] LC-MS: m / z = 219.9 [M+H] + .

[0498] Step 6: Preparation of N-methyl-5-((5-oxo-4,5-dihydrofuro[2,3-d]thieno[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (18).

[0499] N-methyl-5-piperazin-1-ylpicolinamide hydrochloride (17.5 mg, 0.068 mmol) and triethylamine (6.90 mg, 0.068 mmol) were dissolved in 1,2-dichloroethane (2 mL) and methanol (2 mL), and reacted at room temperature for 30 minutes. Acetic acid (0.05 mL) and 5-oxo-4,5-dihydrofurano[2,3-d]thieno[3,2-b]pyridine-2-carbaldehyde (13.6 mg, 0.062 mmol) were added, and reacted at room temperature for 30 minutes. Sodium cyanoborohydride (7.80 mg, 0.124 mmol) was added, and reacted at room temperature for 16 hours. The residue was concentrated under reduced pressure and separated by high pressure preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid) to obtain 4.5 mg of the title compound as a white solid in a yield of 17.1%.

[0500] LC-MS: m / z = 423.9 [M+H] + .

[0501] 1 H NMR (400MHz, DMSO-d6) δ12.06(s,1H),8.41(p,J=5.3,4.9Hz,1H),8.27(d,J=2.8Hz,1H),7.93(d,J=2.1Hz,1H),7.83(d,J=8.7Hz ,1H),7.40(dd,J=8.8,2.9Hz,1H),7.06–6.98(m,2H),3.86(s,2H),3.39–3.34(m,4H),2.78(d,J=4.8Hz,3H),2.67-2.62(m,4H).

[0502] Example 19: Preparation of N-methyl-5-((5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (19).

[0503]

[0504] Step 1: Preparation of methyl 1-(5-methoxycarbonyl)-3-nitrothiophen-2-yl)-1H-pyrrole-2-carboxylate (19a).

[0505] 5-Chloro-4-nitrothiophene-2-carboxylic acid methyl ester (2.22 g, 10.0 mmol) and cesium carbonate (4.07 g, 12.5 mmol) were dissolved in N,N-dimethylformamide (25 mL), 1H-pyrrole-2-carboxylic acid methyl ester (1.50 g, 12.0 mmol) was added, and stirred at 60°C for 16 hours. Water (100 mL) was added for dilution, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-20%) to obtain 2.54 g of the title compound as a yellow solid with a yield of 82.0%.

[0506] LC-MS: m / z = 310.9 [M+H] + .

[0507] Step 2: Preparation of methyl 5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazine-2-carboxylate (19b).

[0508] At room temperature and under nitrogen atmosphere, 1-(5-methoxycarbonyl)-3-nitrothiophen-2-yl)-1H-pyrrole-2-carboxylic acid methyl ester (2.49 g, 8.03 mmol) was dissolved in acetic acid (5 mL), and ethanol (12 mL) and water were added.

[0509] (6mL), zinc powder (2.10g, 32.1mmol), react at 80°C for 16 hours. Filter, wash the filter cake with methanol (30mL), concentrate the filtrate under reduced pressure, and separate and purify the residue by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-25%) to obtain 141mg of the title compound as a yellow solid, with a yield of 7.1%.

[0510] LC-MS: m / z = 248.9 [M+H] + .

[0511] Step 3: Preparation of 2-hydroxymethylpyrrolo[1,2-a]thieno[3,2-e]pyrazin-5(4H)one (19c).

[0512] At 0°C, under nitrogen atmosphere, methyl 5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazine-2-carboxylate (121 mg, 0.485 mmol) was dissolved in tetrahydrofuran (5.0 mL), and a 1.0 M solution of lithium aluminum hydride in tetrahydrofuran (1.94 mL, 1.94 mmol) was added dropwise, and the mixture was reacted at room temperature for 2 hours. Sodium sulfate decahydrate (500 mg) was added to quench the reaction, and the mixture was filtered. The filter cake was washed with methanol (30 mL), and the filtrate was concentrated. The residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-5%) to obtain 97.2 mg of the title compound as a yellow solid, with a yield of 91.0%.

[0513] LC-MS: m / z = 220.9 [M+H] + .

[0514] Step 4: Preparation of 5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazine-2-carbaldehyde (19d).

[0515] At 0°C under nitrogen atmosphere, 2-hydroxymethylpyrrolo[1,2-a]thieno[3,2-e]pyrazin-5(4H)one was added.

[0516] (30.0 mg, 0.136 mmol) was dissolved in dichloromethane (10 mL), and Dess-Martin periodinane (115.4 mg, 0.272 mmol) was added, and the mixture was reacted at 40°C for 16 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-4%) to obtain 57.6 mg of the crude title compound as a yellow solid.

[0517] LC-MS: m / z = 218.9 [M+H] + .

[0518] Step 5: Preparation of N-methyl-5-((5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (19).

[0519] N-Methyl-5-piperazin-1-ylpicolinamide hydrochloride (51.3 mg, 0.200 mmol) was dissolved in 1,2-dichloroethane (5 mL) and methanol (5 mL), triethylamine (20.2 mg, 0.200 mmol) was added, and the mixture was stirred at room temperature for 30 minutes. Acetic acid (0.1 mL) and 5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazine-2-carbaldehyde (39.4 mg, 0.182 mmol) were added, and the mixture was stirred at room temperature for 30 minutes. Sodium cyanoborohydride (22.9 mg, 0.364 mmol) was added, and the mixture was stirred at 70°C for 16 hours. The residue was concentrated under reduced pressure and separated by high pressure preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid) to obtain 6.0 mg of the title compound as a white solid in a yield of 7.8%.

[0520] LC-MS: m / z = 422.9 [M+H] + .

[0521] 1 H NMR (400MHz, DMSO-d6) δ11.39(s,1H),8.41(q,J=4.8Hz,1H),8.27(d,J=3.0Hz,1H),7.83(d,J=8.8Hz,1H),7.66–7.60(m,1H),7.40(dd,J=8.9,2.9Hz ,1H),7.00(dd,J=4.0,1.4Hz,1H),6.83(s,1H),6.62(dd,J=4.0,2.6Hz,1H ),3.79(s,2H),3.36(s,4H),2.77(d,J=4.8Hz,3H),2.63(t,J=5.0Hz,4H).

[0522] Example 20: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (20).

[0523]

[0524] Step 1: Preparation of ethyl 1-(5-(methoxycarbonyl)-3-nitrothiophen-2-yl)-1H-imidazole-2-carboxylate (20a).

[0525] To a solution of methyl 5-chloro-4-nitrothiophene-2-carboxylate (2.00 g, 9.02 mmol) and ethyl 1H-imidazole-2-carboxylate (1.52 g, 10.8 mmol) in N,N-dimethylformamide (20 mL) was added cesium carbonate (3.68 g, 11.3 mmol) at room temperature, and the mixture was stirred at 60°C for 5 hours. The mixture was filtered, the filtrate was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 25%-40%) to obtain 773 mg of the title compound as a yellow solid, with a yield of 26.3%.

[0526] LC-MS: m / z=326.0[M+H] + .

[0527] Step 2: Preparation of ethyl 1-(3-amino-5-(methoxycarbonyl)thiophen-2-yl)-1H-imidazole-2-carboxylate (20b).

[0528] Triphenyl phosphite (74.0 mg, 239 μmol) and ammonium metavanadate (28.0 mg, 239 μmol) were added to a solution of ethyl 1-(5-(methoxycarbonyl)-3-nitrothiophene-2-yl)-1H-imidazole-2-carboxylate (778 mg, 2.39 mmol) in dichloromethane / methanol (v:v=5:1, 18 mL) at room temperature, and 5% platinum carbon (289 mg) was added, and the mixture was stirred at room temperature for 16 hours under an atmospheric pressure of hydrogen. The mixture was filtered, the filtrate was concentrated, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane=0%-10%) to obtain 1.24 g of the title compound as a yellow solid.

[0529] LC-MS: m / z = 296.1 [M+H] + .

[0530] Step 3: Preparation of methyl 5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazine-2-carboxylate (20c).

[0531] Dissolve 1-(3-amino-5-(methoxycarbonyl)thiophen-2-yl)-1H-imidazole-2-carboxylic acid ethyl ester (1.24 g, 4.20 mmol) and potassium carbonate (1.16 g, 8.40 mmol) in N,N-dimethylformamide (18 mL) and stir at 80°C for 16 hours. Add water (250 mL) to dilute, stir at room temperature for 30 minutes, filter, and separate and purify the filter cake by silica gel column chromatography (mobile phase: methanol / dichloromethane = 10%-25%) to obtain 190 mg of the title compound as a yellow solid, yield: 20.0%.

[0532] LC-MS: m / z = 250.0 [M+H] + .

[0533] Step 4: Preparation of 2-(hydroxymethyl)imidazo[1,2-a]thieno[3,2-e]pyrazin-5(4H)-one (20d).

[0534] Dissolve 5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazine-2-carboxylic acid methyl ester (190 mg, 762 μmol) in tetrahydrofuran (6 mL), slowly add 1M lithium aluminum hydride tetrahydrofuran solution (1.22 mL, 1.22 mmol) dropwise at -40°C, and stir at room temperature for 2 hours. Add sodium sulfate decahydrate (200 mg) to quench the reaction, dilute with methanol (50 mL), filter, concentrate the filtrate, and separate and purify the residue by silica gel column chromatography (methanol / dichloromethane = 10%-25%) to obtain 280 mg of the title compound as a yellow solid.

[0535] LC-MS: m / z = 222.0 [M+H] + .

[0536] Step 5: Preparation of 5-oxo-4,5-dihydroimidazo[1,2-a]thiophene[3,2-e]pyrazine-2-carbaldehyde (20e).

[0537] At room temperature, manganese dioxide (2.20 g, 25.3 mmol) was added to a solution of 2-(hydroxymethyl)imidazo[1,2-a]thieno[3,2-e]pyrazin-5(4H)-one (280 mg, 1.27 mmol) in ethyl acetate / methanol (v:v=1:2, 12 mL), and the mixture was reacted at room temperature for 16 hours. The solid was filtered and concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane=10%-25%) to obtain 62.0 mg of the title compound as a yellow oil, yield: 22.4%.

[0538] LC-MS: m / z = 220.0 [M+H] + .

[0539] Step 6: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (20).

[0540] To a solution of N-methyl-5-(piperazin-1-yl)picolinamide (62.0 mg, 283 μmol) in dichloroethane / methanol (v:v=1:1, 12.0 mL) was added triethylamine (40 μL, 283 μmol) at room temperature, and the mixture was stirred at room temperature for 15 minutes. Glacial acetic acid (80 μL, 1.41 mmol) and 5-oxo-4,5-dihydroimidazo[1,2-a]thienyl[3,2-e]pyrazine-2-carbaldehyde (250 mg, 392 μmol) were added, and the mixture was stirred at room temperature for 16 hours. Sodium cyanoborohydride (27.0 mg, 424 μmol) was added, and the mixture was stirred at 50°C for 1 hour. The residue was concentrated under reduced pressure and separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 3.0 mg of the title compound as a white solid. Yield: 2.5%.

[0541] LC-MS: m / z = 423.9 [M+H] + .

[0542] 1 H NMR (400MHz, DMSO-d6) δ12.01(s,1H),8.40(q,J=4.8Hz,1H),8.27(d,J=2.8Hz,1H),8.11(d,J=1.2Hz,1H),7.83(d,J=8.8Hz,1H),7.5 5(d,J=1.2Hz,1H),7.40(dd,J=8.8,2.8Hz,1H),6.90(s,1H),3.82(s,2H),3.31(s,4H),2.78(d,J=4.8Hz,3H),2.65(t,J=5.2Hz,4H).

[0543] Example 21: Preparation of N-methyl-1′-((5-oxo-5,6,7,8-tetrahydro-4H-cyclopentylthieno[3,2-b]pyridin-2-yl)methyl)-1,2,3,6-tetrahydro-[3,4′-bipyridine]-6-carboxamide (21).

[0544]

[0545] Step 1: Preparation of methyl 1-tert-butyl-6-methyl-3,6-dihydrobipyridine-1,6(2'H)dicarboxylate (21a).

[0546] At room temperature and under nitrogen atmosphere, methyl 5-bromopicolinate (2.16 g, 10.0 mmol) and tert-butyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3,6-dihydropyridine-1(2H)-carboxylate (3.71 g, 12.0 mmol) were dissolved in 1,4-dioxane (30 mL) and water (7.5 mL). [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (732 mg, 1.00 mmol) and potassium carbonate (2.76 g, 20.0 mmol) were added, and the mixture was stirred at 80°C for 3 hours. Water (50 mL) was added to dilute, and the mixture was extracted with ethyl acetate (50 mL x 3). The organic phase was washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-20%) to obtain 3.18 g of the title compound as a yellow solid. The yield was 99.0%.

[0547] LC-MS: m / z 319.2[M+H] + .

[0548] Step 2: Preparation of tert-butyl 6-methylcarbamoyl-3,6-dihydro-[3,4-bipyridine]-1-carboxylate (21b).

[0549] At room temperature, 1-tert-butyl-6-methyl-3,6-dihydrobipyridine-1,6(2'H)dicarboxylic acid methyl ester (1.50 g, 4.71 mmol) was dissolved in methanol (15 mL), and a 40% aqueous methylamine solution (3.66 g, 47.1 mmol) was added, and the mixture was reacted at room temperature for 16 hours. The mixture was concentrated under reduced pressure, diluted with water (20 mL), and extracted with ethyl acetate (30 mL x 3). The organic phase was washed with a saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-25%) to obtain 1.38 g of the title compound as a yellow oil, with a yield of 92.4%.

[0550] LC-MS: m / z=318.3[M+H] + .

[0551] Step 3: Preparation of N-methyl-1,2,3,6-tetrahydrobipyridine-6-carboxamide hydrochloride (21c).

[0552] At room temperature, tert-butyl 6-methylcarbamoyl-3,6-dihydro-[3,4-bipyridine]-1-carboxylate (1.38 g, 4.35 mmol) was dissolved in methanol (15 mL), 4M hydrochloric acid dioxane solution (5.9 mL, 23.5 mmol) was added, and the mixture was reacted at room temperature for 16 hours. The mixture was concentrated under reduced pressure to obtain 1.41 g of the title compound as a white solid.

[0553] LC-MS: m / z = 218.0 [M+H] + .

[0554] Step 4: Preparation of N-methyl-1'-((5-oxo-5,6,7,8-tetrahydro-4H-cyclopentylthieno[3,2-b]pyridin-2-yl)methyl)-1,2,3,6-tetrahydro-[3,4'-bipyridine]-6-carboxamide (21).

[0555] At room temperature and under nitrogen atmosphere, 2-chloromethyl-4,6,7,8-tetrahydro-5H-cyclopentyl[d]thieno[3,2-b]pyridin-5-one (400 mg, 1.67 mmol) and N-methyl-1,2,3,6-tetrahydrobipyridine-6-carboxamide hydrochloride (467 mg, 1.84 mmol) were dissolved in acetonitrile (10 mL), and N,N-diisopropylethylamine (1.30 g, 10.0 mmol) and sodium iodide (501 mg, 3.34 mmol) were added, and the mixture was reacted at 50°C for 16 hours. The residue was concentrated under reduced pressure and separated by high pressure preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid) to obtain 40.0 mg of the title compound as a yellow solid in a yield of 5.7%.

[0556] LC-MS: m / z = 420.9 [M+H] + .

[0557] 1 H NMR(400MHz, Methanol-d4)δ8.86(d,J=2.1Hz,1H),8.37(dd,J=8.3,2.2Hz,1H),8.25(d,J=8.3Hz,1H),7.69(s,1H), 6.48(s,1H),4.92(s,4H),4.10(s,2H),3.21(t,J=7.6Hz,2H),3.15–3.01(m,4H),3.00(s,3H),2.33(p,J=7.7Hz,2H)

[0558] Example 22: Preparation of N-methyl-5-((6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (22).

[0559]

[0560] Step 1: Preparation of methyl 1-(5-methoxycarbonyl)-3-nitrothiophen-2-yl)-3-methylpyrrole-2-carboxylate (22a).

[0561] At room temperature and under nitrogen atmosphere, 5-chloro-4-nitrothiophene-2-carboxylic acid methyl ester (2.22 g, 10.0 mmol) was dissolved in N,N-dimethylformamide (25 mL), and 3-methylpyrrole-2-carboxylic acid methyl ester (1.84 g, 12.0 mmol) and cesium carbonate (4.07 g, 12.5 mmol) were added, and stirred at 60°C for 16 hours. Water (100 mL) was added for dilution, and the mixture was extracted with ethyl acetate (100 mL x 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate / petroleum ether = 1%-20%) to obtain 3.20 g of the title compound as a yellow solid with a yield of 94.5%.

[0562] LC-MS: m / z 338.9[M+H] + .

[0563] Step 2: Preparation of methyl 6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazine-2-carboxylate (22b).

[0564] At room temperature and under nitrogen atmosphere, 1-(5-methoxycarbonyl)-3-nitrothiophen-2-yl)-3-methylpyrrole-2-carboxylic acid methyl ester (3.10 g, 9.15 mmol) was dissolved in acetic acid (30 mL), iron powder (2.05 g, 36.6 mmol) was added, and the mixture was stirred at 80°C for 2 hours. The mixture was filtered, and the filter cake was washed with methanol (100 mL). The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-5%) to obtain 995 mg of the title compound as a yellow solid, with a yield of 41.5%.

[0565] LC-MS: m / z = 262.9 [M+H] + .

[0566] Step 3: Preparation of 2-hydroxymethyl-6-methylpyrrolo[1,2-a]thieno[3,2-e]pyrazin-5(4H)one (22c).

[0567] At 0°C under nitrogen atmosphere, 6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]

[0568] Pyrazine-2-carboxylic acid methyl ester (895 mg, 3.41 mmol) was dissolved in tetrahydrofuran (20 mL), and a 2.5 M solution of lithium aluminum hydride in tetrahydrofuran (2.73 mL, 6.82 mmol) was added dropwise, and stirred at room temperature for 2 hours. Sodium sulfate decahydrate was added to quench the reaction, and the mixture was filtered, and the filter cake was washed with methanol (50 mL). The filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-6%) to obtain 409 mg of the title compound as a yellow solid, with a yield of 51.2%.

[0569] LC-MS: m / z = 234.9 [M+H] + .

[0570] Step 4: Preparation of 6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazine-2-carbaldehyde (22d).

[0571] At 0°C, under nitrogen atmosphere, 2-hydroxymethyl-6-methylpyrrolo[1,2-a]thieno[3,2-e]pyrazin-5(4H)one (359 mg, 1.53 mmol) was dissolved in dichloromethane (20 mL), and Dess-Martin periodinane (1.30 mg, 3.06 mmol) was added, and stirred at 40°C for 16 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol / dichloromethane = 1%-5%) to obtain 561 mg of the crude title compound as a yellow solid.

[0572] LC-MS: m / z = 232.9 [M+H] + .

[0573] Step 5: Preparation of N-methyl-5-((6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (22).

[0574] At room temperature and under nitrogen atmosphere, N-methyl-5-piperazin-1-ylpicolinamide hydrochloride (305.5 mg, 1.19 mmol) was dissolved in 1,2-dichloroethane (10 mL) and methanol (10 mL). Triethylamine (120 mg, 1.19 mmol) was added and stirred at room temperature for 30 minutes. Acetic acid (0.4 mL) and 6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazine-2-carbaldehyde (250 mg, 1.08 mmol) were added and stirred at room temperature for 30 minutes. Sodium cyanoborohydride (136 mg, 2.16 mmol) was added and stirred at 70°C for 16 hours. The residue was concentrated under reduced pressure and separated by high pressure preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid) to obtain 141 mg of the title compound as a yellow solid in a yield of 29.9%.

[0575] LC-MS: m / z = 436.9 [M+H] + .

[0576] 1 H NMR (400MHz, DMSO-d6) δ11.35(s,1H),8.45(q,J=4.8Hz,1H),8.33(d,J=2.9Hz,1H),7.88(d,J=8.7Hz,1H),7.58(d,J=2.7Hz,1H),7.48(dd,J= 8.8,2.9Hz,1H),7.11(s,1H),6.49(d,J=2.6Hz,1H),4.64(s,2H),4.09(s,2H),3.49(s,2H),3.18(s,4H),2.78(d,J=4.7Hz,3H),2.49(s,3H).

[0577] Example 23: Preparation of N-methyl-5-(4-((1-methyl-5-oxyl-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinolin-2-yl)methyl)piperazin-1-yl)picolinamide (23).

[0578]

[0579] Step 1: Preparation of 1-methyl-4-nitro-1H-pyrrole-2-carbaldehyde (23a).

[0580] At 0°C, 1-methyl-1H-pyrrole-2-carboxaldehyde (1.00 g, 9.17 mmol) was dissolved in 2.50 mL H2SO4. Nitric acid (0.50 ml) was slowly added dropwise under stirring at 0°C. The reaction was terminated after 1 min. The reaction solution was carefully poured into ice. EA was extracted (50 mL x 3), washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA = 1:0-0:1) to obtain 800 mg of the title compound as a dark yellow solid, with a yield of 55.4%.

[0581] LC-MS: m / z=155[M+H] + .

[0582] Step 2: Preparation of 5-bromo-1-methyl-4-nitro-1H-pyrrole-2-carbaldehyde (23b).

[0583] 1-Methyl-4-nitro-1H-pyrrole-2-carboxaldehyde (3.80 g, 24.7 mmol) was dissolved in 38.0 mL DMF, Br2 (4.74 g, 29.6 mmol) was added, and the mixture was stirred at 100°C overnight. 200 mL of water was added to quench the mixture, and the mixture was extracted with EA (100 mL x 3), washed with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by TLC chromatography (mobile phase: PE / EA = 10:1) to obtain 2.00 g of the title compound as a light yellow solid, with a yield of 34.9%.

[0584] LC-MS: m / z=233[M+H] + .

[0585] Step 3: Preparation of methyl 2-(5-formyl-1-methyl-3-nitro-1H-pyrrol-2-yl)benzoate (23c).

[0586] 5-Bromo-1-methyl-4-nitro-1H-pyrrole-2-carboxaldehyde (200 mg, 0.86 mmol) was dissolved in 5 mL of dioxane and 0.5 mL of water, and methyl 2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (271 mg, 1.03 mmol), K2CO3 (237 mg, 1.72 mmol), Pd(dppf)Cl2 (31.4 mg, 0.043 mmol) were added, and stirred at 110°C overnight under nitrogen atmosphere. 20 mL of water was added to quench, EA was extracted (30 mL x 3), washed with saturated brine (20 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by high pressure preparative liquid phase separation (chromatographic column model: Daisogei

[0587] 30mm*250mm, C18, 10um 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%), to obtain 100 mg of the title compound as a dark grey solid, yield: 40.3%.

[0588] LC-MS: m / z=289[M+H] + .

[0589] Step 4: Preparation of 1-methyl-5-oxo-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinoline-2-carbaldehyde (23d).

[0590] Methyl 2-(5-formyl-1-methyl-3-nitro-1H-pyrrol-2-yl)benzoate (80 mg, 0.28 mmol) was dissolved in 2.5 mL of AcOH, and Fe powder (77.8 mg, 1.40 mmol) was added, and stirred at 80°C overnight. 10 mL of water was added to quench, and saturated NaHCO3 solution was used to adjust pH>7, and EA was extracted (20 mL x 3), washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 64.0 mg of the crude title compound as a gray solid, with a yield of 100%.

[0591] LC-MS: m / z 227[M+H] + .

[0592] Step 5: Preparation of N-methyl-5-(4-((1-methyl-5-oxy-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinolin-2-yl)methyl)piperazin-1-yl)picolinamide (23).

[0593] Dissolve N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride (73.4 mg, 0.29 mmol) in 5.0 mL of THF, add DIPEA (61.9 mg, 0.48 mmol), stir at room temperature for 0.5 h, add 1-methyl-5-oxo-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinoline-2-carbaldehyde (54.0 mg, 0.24 mmol), Ti(OEt)4 (137 mg, 0.60 mmol), stir at room temperature for 1 h, add NaBH3CN (30.1 mg, 0.48 mmol), MeOH (1.00 ml), stir at room temperature overnight. The mixture was quenched by adding 20 mL of saturated aqueous sodium carbonate solution, extracted with EA (10 mL x 3), washed with saturated brine (20.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by high pressure preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to give 13.0 mg of the title compound as a white solid. Yield: 12.6%.

[0594] LC-MS: m / z=431[M+H] + .

[0595] 1 H NMR (400MHz, DMSO) δ = 11.42 (s, 1H), 8.37 (dd, 1H), 8.31 (q, 1H), 8.25 (d, 1H), 8.17 (d, 1H), 7.82 (d, 1H), 7.74 -7.70 (m, 1H), 7.39 -7.35(m,2H),5.99(s,1H),4.05(s,3H),3.54(s,2H),2.78(d,3H),2.56(t,4H),2.51–2.50(m,4H).

[0596] Example 24: Preparation of 5-(4-((8-fluoro-1-methyl-5-oxy-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinolin-2-yl)methyl)piperazin-1-yl)N-methylpicolinamide (24).

[0597]

[0598] Step 1: Preparation of methyl 4-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (24a).

[0599] At 0°C, methyl 2-bromo-4-fluorobenzoate (5.00 g, 21.5 mmol) was dissolved in 70.0 mL of Dio, and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxaborolane) (6.54 g, 25.8 mmol), KOAc (4.21 g, 43.0 mmol), and Pd(dppf)Cl2 (787 mg, 1.08 mmol) were added. The mixture was stirred at 110°C overnight under N2 atmosphere. The mixture was quenched by adding 100 mL of water, extracted with EA (100 mL x 3), washed with saturated brine (20.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: PE / EA=1:0-0:1) to obtain 3.70 g of the title compound as a brown oil. Yield: 61.6%.

[0600] LC-MS: m / z=282[M+H] + .

[0601] Step 2: Preparation of methyl 4-fluoro-2-(5-formyl-1-methyl-3-nitro-1H-pyrrol-2-yl)benzoate (24b).

[0602] 5-Bromo-1-methyl-4-nitro-1H-pyrrole-2-carbaldehyde (200 mg, 0.86 mmol) was dissolved in 5.00 / 0.50 mL Dio / H2O, and methyl 4-fluoro-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzoate (288 mg, 1.03 mmol), K2CO3 (237 mg, 1.72 mmol), and Pd(dppf)Cl2 (31.4 mg, 0.043 mmol) were added, and the mixture was stirred at 110°C overnight under N2 atmosphere. The mixture was quenched by adding 20 mL of water, extracted with EA (30 mL x 3), washed with saturated brine (20.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated by high-pressure preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to give 100 mg of the title compound as a dark gray solid. Yield: 38.0%.

[0603] LC-MS: m / z=308[M+H] + .

[0604] Step 3: Preparation of 8-fluoro-1-methyl-5-oxo-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinoline-2-carbaldehyde (24c).

[0605] Methyl 4-fluoro-2-(5-formyl-1-methyl-3-nitro-1H-pyrrol-2-yl)benzoate (80 mg, 0.26 mmol) was dissolved in 2.50 mL of AcOH, and Zn powder (85.0 mg, 1.31 mmol) was added, and stirred at 80°C overnight. 10 mL of water was added to quench, saturated NaHCO3 solution was used to adjust pH>7, EA was extracted (20 mL x 3), washed with saturated brine (10.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: DCM / MeOH=1:0-0:1) to obtain 16.0 mg of the title compound as a brown solid, with a yield of 25.3%.

[0606] LC-MS: m / z=245[M+H] + .

[0607] Step 4: Preparation of 5-(4-((8-fluoro-1-methyl-5-oxy-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinolin-2-yl)methyl)piperazin-1-yl)N-methylpicolinamide (24).

[0608] Dissolve N-methyl-5-(piperazin-1-yl)picolinamide hydrochloride (76.8 mg, 0.30 mmol) in 1.00 ml THF, add 8-fluoro-1-methyl-5-oxo-4,5-dihydro-1H-pyrrolo[3,2-c]isoquinoline-2-carbaldehyde (60.0 mg, 0.25 mmol), Ti(OEt)4 (114 mg, 0.50 mmol), stir at room temperature for 1 hour, add NaBH3CN (31.4 mg, 0.50 mmol), MeOH (0.50 ml), stir at room temperature overnight. The mixture was quenched by adding 20 mL of saturated aqueous sodium carbonate solution, extracted with EA (10 mL x 3), washed with saturated brine (20.0 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by TLC chromatography (mobile phase: DCM / MeOH=10:1), and then separated by high-pressure preparative liquid separation (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water (0.05% formic acid), gradient: 30%-70%) to give 3.50 mg of the title compound as a white solid, in a yield of 3.12%.

[0609] LC-MS: m / z=449[M+H] + .

[0610] 1H NMR (400MHz, DMSO) δ = 11.47 (s, 1H), 8.38 -8.33 (m, 2H), 8.26 (d, 1H), 7.86 -7.81(m,2H),7.38(dd,1H),7.18(ddd,1H),6.00(s,1H),4.03(s,3H),3.64(s,2H),2.77(d,3H),2.56(t,4H),2.51–2.50(m,4H).

[0611] Example 25: Preparation of 5-(4-(6-fluoro-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (25).

[0612]

[0613] Referring to the preparation method of Example 22, 3-methylpyrrole-2-carboxylic acid methyl ester in step 1 was replaced by 3-fluoro-1H-pyrrole-2-carboxylic acid methyl ester.

[0614] LC-MS: m / z = 440.9 [M+H] + .

[0615] 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.45(q,J=4.8Hz,1H),8.33(d,J=2.9Hz,1H),7.89(d,J=8.7Hz,1H),7.65(t,J=3.7Hz,1H) ,7.48(dd,J=8.8,2.9Hz,1H),7.13(s,1H),6.60(d,J=3.1Hz,1H),4.64(s,2H),4.09(s,2H),3.33(m,,6H),2.79(d,J=4.7Hz,3H).

[0616] Example 26: Preparation of N-methyl-5-(4-(1-methyl-5-oxo-1,4,5,6,7,8-hexahydrocyclopentyl)pyrrolo[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (26).

[0617]

[0618] Referring to the preparation method of Example 17, 5-bromo-4-nitrothiophene-2-carboxylic acid methyl ester in step 3 was replaced by 1-methyl-4-nitro-1H-pyrrole-2-carboxylic acid methyl ester.

[0619] LC-MS: m / z 421.0[M+H]+ .

[0620] 1 H NMR (400MHz, DMSO-d6) δ11.53(s,1H),8.45(q,J=4.8Hz,1H),8.33(d,J=2.9Hz,1H),7.89(d,J=8.7Hz,1H),7.47(dd,J=8.8,2.9Hz,1H),6.26(s, 1H),4.55(s,2H),4.09(s,2H),3.84(s,3H),3.48(s,2H),3.42–3.05(m, 6H), 2.79 (d, J = 4.7Hz, 3H), 2.66 (t, J = 7.4Hz, 2H), 2.07 (p, J = 7.6Hz, 2H).

[0621] Example 27: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (27).

[0622]

[0623]

[0624] Step 1: Preparation of ethyl 1-(5-(methoxycarbonyl)-3-nitrothiophen-2-yl)-1H-imidazole-2-carboxylate (27a).

[0625] To a solution of methyl 5-chloro-4-nitrothiophene-2-carboxylate (2.00 g, 9.02 mmol) and ethyl 1H-imidazole-2-carboxylate (1.52 g, 10.8 mmol) in N,N-dimethylformamide (20.0 mL) was added cesium carbonate (3.68 g, 11.3 mmol) at room temperature, and the mixture was stirred at 60°C for 5 hours. The mixture was filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether:ethyl acetate = 25%-40%) to obtain 773 mg of the title compound as a yellow solid, with a yield of 26.3%.

[0626] LC-MS: m / z=326[M+H] + .

[0627] Step 2: Preparation of ethyl 1-(3-amino-5-(methoxycarbonyl)thiophen-2-yl)-1H-imidazole-2-carboxylate (27b).

[0628] Triphenyl phosphite (74.0 mg, 239 μmol) and ammonium metavanadate (28.0 mg, 239 μmol) were added to a solution of ethyl 1-(5-(methoxycarbonyl)-3-nitrothiophene-2-yl)-1H-imidazole-2-carboxylate (778 mg, 2.39 mmol) in dichloromethane and methanol (18 mL, dichloromethane / methanol = 5:1) at room temperature. The reaction system was replaced with nitrogen three times, 5% Pt / C (289 mg) was added and replaced with hydrogen three times, and stirred at room temperature for 16 hours in a hydrogen atmosphere (15 Psi). The mixture was filtered and concentrated under reduced pressure. The residual compound was separated and purified by silica gel column chromatography (mobile phase: dichloromethane:methanol = 0%-10%) to obtain 1.24 g of the title compound as a yellow solid.

[0629] LC-MS: m / z=296[M+H] + .

[0630] Step 3: Preparation of methyl 5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazine-2-carboxylate (27c).

[0631] Potassium carbonate (1.16 g, 8.40 mmol) was added to a solution of ethyl 1-(3-amino-5-(methoxycarbonyl)thiophen-2-yl)-1H-imidazole-2-carboxylate (1.24 g, 4.20 mmol) in N,N-dimethylformamide (18 mL) at room temperature, and stirred at 80°C for 16 hours. The mixture was diluted with water (250 mL), stirred at room temperature for 30 minutes, filtered, and the filter cake was collected and dried under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: dichloromethane: methanol = 10%-25%) to obtain 190 mg of the title compound as a yellow solid, with a yield of 20.0%. .

[0632] LC-MS: m / z = 250 [M+H] + .

[0633] Step 4: Preparation of 2-(hydroxymethyl)imidazo[1,2-a]thieno[3,2-e]pyrazin-5(4H)-one (27d).

[0634] To a solution of methyl 5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazine-2-carboxylate (190 mg, 762 μmmol) in tetrahydrofuran (6.0 mL) was added lithium aluminum hydride (1.22 mL, 1.22 mmol, 1 M in THF) at room temperature, and the mixture was stirred at room temperature for 2 hours. Sodium sulfate decahydrate (200 mg) was added to the reaction solution, and the mixture was diluted with methanol (50 mL), filtered, and the filtrate was concentrated. The residue was separated and purified by silica gel column chromatography (dichloromethane:methanol=10%-25%) to obtain 280 mg of the title compound as a yellow solid.

[0635] LC-MS: m / z=222[M+H] + .

[0636] Step 5: Preparation of 5-oxo-4,5-dihydroimidazo[1,2-a]thiophene[3,2-e]pyrazine-2-carbaldehyde (27e).

[0637] At room temperature, add manganese dioxide (2.20 g, 25.3 mmol) to a mixed solution of 2-(hydroxymethyl)imidazo[1,2-a]thieno[3,2-e]pyrazin-5(4H)-one (280 mg, 1.27 mmol) in ethyl acetate and methanol (126 mL, ethyl acetate / methanol = 1:2), and stir at room temperature for 16 hours. Filter, concentrate the filtrate, and separate and purify the residue by silica gel column chromatography (mobile phase: dichloromethane:methanol = 10%-25%) to obtain 62.0 mg of the title compound as a yellow oil, yield: 22.4%.

[0638] LC-MS: m / z = 220 [M+H] + .

[0639] Step 6: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydroimidazo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (27).

[0640] To a mixed solution of N-methyl-5-(piperazin-1-yl)picolinamide (62.0 mg, 283 μmol) in dichloroethane and methanol (12.0 mL, dichloroethane / methanol = 1:1) was added triethylamine (39.0 μL, 283 μmol) at room temperature. After stirring at room temperature for 15 minutes, glacial acetic acid (80.1 μL, 1.41 mmol) and 5-oxo-4,5-dihydroimidazo[1,2-a]thiophene[3,2-e]pyrazine-2-carbaldehyde (250 mg, 392 μmol, 30%) were added. The mixture was stirred for 16 hours. Sodium cyanoborohydride (27.0 mg, 424 μmol) was added and stirred at 50°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the residual compound was separated by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 3.0 mg of the title compound as a white solid. Yield: 2.5%.

[0641] LC-MS: m / z=423.90 [M+H] + .

[0642] 1H NMR (400MHz, DMSO-d6) δ12.01(s,1H),8.40(q,J=4.8Hz,1H),8.27(d,J=2.8Hz,1H),8.11(d,J=1.2Hz,1H),7.83(d,J=8.8Hz,1H),7.5 5(d,J=1.2Hz,1H),7.40(dd,J=8.8,2.8Hz,1H),6.90(s,1H),3.82(s,2H),3.31(s,4H),2.78(d,J=4.8Hz,3H),2.65(t,J=5.2Hz,4H).

[0643] Example 28: Preparation of 5-(4-((6-chloro-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (28).

[0644]

[0645] Referring to the preparation method of Example 22, 3-methylpyrrole-2-carboxylic acid methyl ester in step 1 was replaced by 3-chloropyrrole-2-carboxylic acid methyl ester.

[0646] LC-MS: m / z=458.1[M+H] + .

[0647] 1 H NMR (400MHz, DMSO-d6) δ11.54(s,1H),8.47(q,J=4.8Hz,1H),8.30(d,J=2.9Hz,1H),7.85(d,J=8.7Hz,1H),7.63(t,J=3.7Hz,1H) ,7.46(dd,J=8.8,2.9Hz,1H),7.11(s,1H),6.65(d,J=3.1Hz,1H),4.66(s,2H),4.09(s,2H),3.31(m,,6H),2.78(d,J=4.7Hz,3H).

[0648] Example 29: Preparation of 5-(4-((6-cyclopropyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (29).

[0649]

[0650] Referring to the preparation method of Example 22, 3-methylpyrrole-2-carboxylic acid methyl ester in step 1 was replaced by 3-cyclopropyl-1H-pyrrole-2-carboxylic acid methyl ester.

[0651] LC-MS: m / z=463.2[M+H] + .

[0652] 1 H NMR (400MHz, DMSO-d6) δ11.35(s,1H),8.45(q,J=4.8Hz,1H),8.33(d,J=2.9Hz,1H ),7.88(d,J=8.7Hz,1H),7.58(d,J=2.7Hz,1H),7.48(dd,J=8.8,2.9Hz,1H),7.11 (s,1H),6.49(d,J=2.6Hz,1H),4.64(s,2H),4.09(s,2H),3.49(s,2H),3.18(s,4H ),2.78(d,J=4.7Hz,3H),2.24-2.18(m,1H),1.32-1.24(m,1H),1.08-0.96(m,1H),

[0653] Example 30: Preparation of N-methyl-5-(4-((3-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (30).

[0654]

[0655] Referring to the preparation method of Example 19, 5-chloro-4-nitrothiophene-2-carboxylic acid methyl ester in step 1 was replaced by methyl 5-chloro-3-methyl-4-nitrothiophene-2-carboxylate.

[0656] LC-MS: m / z=437.2[M+H] + .

[0657] 1 H NMR (400MHz, DMSO-d6) δ11.36(s,1H),8.42(q,J=4.8Hz,1H),8.24(d,J=3.0Hz,1H),7.86(d,J=8.8Hz,1H),7.66–7.60(m,1H),7.41(dd,J=8.9,2.9Hz ,1H),7.05(dd,J=4.0,1.4Hz,1H),6.67(dd,J=4.0,2.6Hz,1H),3.78(s,2H ),3.34(s,4H),2.78(d,J=4.8Hz,3H),2.62(t,J=5.0Hz,4H),2.28(s,3H).

[0658] Example 31: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydropyrazolo[1,5-a]pyrido[3,2-e]pyrimidin-2-yl)methyl)piperazin-1-yl)picolinamide (31).

[0659]

[0660] Step 1: Preparation of ethyl 5-oxo-4,5-dihydropyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-2-carboxylate (31a).

[0661] At -10°C, a solution of 2-chloronicotinoyl chloride (1.13 g, 6.45 mmol) in N, N-dimethylformamide (2.0 mL) was added dropwise to a solution of ethyl 5-amino-1H-pyrazole-3-carboxylate (1.00 g, 6.45 mmol) and potassium carbonate (1.78 g, 12.9 mmol) in N, N-dimethylformamide (8.0 mL), and the mixture was stirred at -10°C for 20 min, and then heated to 140°C and stirred for 30 min. The reaction solution was stirred for 20 minutes with ethyl acetate: petroleum ether = 1:1 (40 mL), filtered, and the filter cake was washed with ethyl acetate: petroleum ether = 1:1 (40 mL x 1), the filter cake was collected, and dried under reduced pressure. The residual compound was separated and purified by silica gel column chromatography (mobile phase: methanol: dichloromethane

[0662] =0%-15%) to give 501 mg of the title compound as a yellow solid, yield: 30.1%.

[0663] LC-MS: m / z = 259.1 [M+H] + .

[0664] Step 2: Preparation of 2-(hydroxymethyl)pyrazolo[1,5-a]pyrido[3,2-e]pyrimidin-5(4H)-one (31b).

[0665] At 0°C, lithium aluminum hydride (2.5M in THF, 836μL, 2.09mmol) was added dropwise to a solution of ethyl 5-oxo-4,5-dihydropyrazolo[1,5-a]pyrido[3,2-e]pyrimidine-2-carboxylate (450mg, 1.74mmol) in tetrahydrofuran (9.0mL), and stirred at 0°C for 1 hour. Methanol (50mL) was added dropwise, filtered, and the filtrate was concentrated under reduced pressure. The residual compound was separated and purified by silica gel column chromatography (mobile phase: methanol: dichloromethane = 0%-15%) to obtain 146mg of the title compound as a yellow solid, yield: 38.8%.

[0666] LC-MS: m / z = 217.1 [M+H] + .

[0667] Step 3: Preparation of 2-(chloromethyl)pyrazolo[1,5-a]pyrido[3,2-e]pyrimidin-5(4H)-one (31c).

[0668] At room temperature, N,N-dimethylformamide (26 μL, 338 μmol) and thionyl chloride (296 μL, 405 μmol) were added to a solution of 2-(hydroxymethyl)pyrazolo[1,5-a]quinazolin-5(4H)-one (146 mg, 675 μmol) in dichloromethane (6.0 mL), and stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: methanol: dichloromethane = 5%-25%) to obtain 103 mg of the title compound as a yellow solid, yield: 78.9%.

[0669] LC-MS: m / z = 235.0 [M+H] + .

[0670] Step 4: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydropyrazolo[1,5-a]pyrido[3,2-e]pyrimidin-2-yl)methyl)piperazin-1-yl)picolinamide (31).

[0671] To a solution of 2-(chloromethyl)pyrazolo[1,5-a]pyrido[3,2-e]pyrimidin-5(4H)-one (100 mg, 426 μmol) and N-methyl-5-(piperazin-1-yl)picolinamide (100 mg, 426 μmol) in acetonitrile (4.0 mL) were added N,N-diisopropylethylamine (371 μL, 2.13 mmol) and sodium iodide (128 mg, 852 μmol) at room temperature and the mixture was stirred at room temperature for 16 hours. After concentration under reduced pressure, the residual compound was separated and purified by silica gel column chromatography (mobile phase: dichloromethane: methanol = 5%-20%) and secondary separation by preparative liquid chromatography (chromatographic column model: Daisogei 30mm*250mm, C18, 10um, 100A, mobile phase: acetonitrile / water, gradient: 30%-80%) to obtain 24.0 mg of the title compound as a white solid. Yield: 13.5%.

[0672] LC-MS: m / z=418.95[M+H] + .

[0673] 1H NMR (400MHz, DMSO-d6) δ12.54(s,1H),8.87(d,J=4.8Hz,1H),8.58(dd,J=8.0,2.0Hz,1H),8.46(t,J=4.8Hz,1H),8.35(d,J=2.8Hz,1H),7.89(d,J=8.8 Hz,1H),7.64(dd,J=7.6,4.8Hz,1H),7.49(dd,J=8.8,2.8Hz,1H),6.19(s,1 H),4.55(s,2H),4.09(s,4H),2.79(d,J=4.8Hz,3H),2.55(d,J=5.2Hz,4H).

[0674] Example 32: Preparation of N-methyl-5-(4-((5-oxo-4,5,6,8-tetrahydrofuro[3,4-d]thieno[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (32).

[0675]

[0676] Referring to the preparation method of Example 17, the 2-oxocyclopentane-1-carboxylic acid methyl ester in step 1 was replaced by 4-oxotetrahydrofuran-3-carboxylic acid methyl ester.

[0677] LC-MS: m / z=426.1[M+H] + .

[0678] 1 H-NMR (400MHz, CDCl3 / CD3OD) δ8.19(d,1H,J=2.8Hz),7.98(d,1H,J=8.8Hz),7.26(dd,1H,J=8.8Hz,J=3.2Hz),7.03(s,1 H), 5.21 (t, 2H, J = 3.6Hz), 5.13 (t, 1H, J = 3.6Hz), 3.87 (s, 2H), 3.40 (t, 4H, J = 4.8Hz), 3.00 (s, 3H), 2.74 (t, 4H, J = 4.8Hz).

[0679] Example 33: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydrodieno[3,2-b:3',2'-d]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (33).

[0680]

[0681] Step 1: Preparation of methyl 3-bromothiophene-2-carboxylate (33a).

[0682] At 0°C, concentrated sulfuric acid (16.5 mL) was added dropwise to a solution of 3-bromothiophene-2-carboxylic acid (5.00 g, 24.2 mmol) in methanol (119 mL), and stirred at 70°C for 16 hours. The mixture was concentrated under reduced pressure, and ethyl acetate (200 mL) and water (100 mL) were added for dilution. The organic phase was washed with saturated sodium bicarbonate solution (100 mL x 1) and saturated sodium chloride solution (100 mL x 1) in sequence, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residual compound was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate: petroleum ether = 15%-35%) to obtain 5.04 g of the title compound as a yellow oil, with a yield of 94.4%.

[0683] LC-MS: m / z = 221.1 [M+H] + .

[0684] Step 2: Preparation of methyl 3-(tributyltinyl)thiophene-2-carboxylate (33b).

[0685] At room temperature, hexabutyl ditin (10.3 mL, 20.4 mmol) and tetrakis(triphenylphosphine) palladium (1.88 g, 1.63 mmol) were added to a toluene (72 mL) solution of methyl 3-bromothiophene-2-carboxylate (3.6 g, 16.4 mmol). The reaction solution was replaced with nitrogen three times and stirred at 110°C for 4 hours. Ethyl acetate (300 mL) was added for dilution, and the mixture was washed with saturated potassium fluoride solution (100 mL x 1), water (100 mL x 1), and saturated sodium chloride solution (100 mL x 1) in sequence. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residual compound was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate: petroleum ether = 0%-5%) to obtain 7.20 g of the title compound as a colorless oil.

[0686] LC-MS: m / z=432.2[M+H] + .

[0687] Step 3: Preparation of dimethyl 3-nitro-[2,3'-dithiophene]-2',5-dicarboxylate (33c).

[0688] At room temperature, tetrakis(triphenylphosphine)palladium (609 mg, 527 μmol) was added to a solution of methyl 3-(tributyltinyl)thiophene-2-carboxylate (5.00 g, 5.80 mmol, 50%) and methyl 5-bromo-4-nitrothiophene-2-carboxylate (1.40 g, 5.27 mmol) in N,N-dimethylformamide (25.0 mL). The reaction solution was replaced with nitrogen three times and stirred at 80°C for 16 hours. The reaction solution was diluted with water (100 mL), extracted with ethyl acetate (100 mL x 2), the organic phases were combined, washed with saturated sodium chloride solution (100 mL x 1), and dried over anhydrous sodium sulfate.

[0689] The residue was filtered and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: ethyl acetate:petroleum ether = 15%-45%) to obtain 270 mg of the title compound as a yellow solid. Yield: 15.7%.

[0690] LC-MS: m / z = 328.3 [M+H] + .

[0691] Step 4: Preparation of methyl 5-oxo-4,5-dihydrodieno[3,2-b:3',2'-d]pyridine-2-carboxylate (33d).

[0692] At room temperature, reduced iron powder (230 mg, 4.12 mmol) was added to a solution of 3-nitro-[2,3'-dithiophene]-2',5-dicarboxylic acid dimethyl ester (270 mg, 825 μmol) in acetic acid (5.0 mL), and stirred at 80°C for 5 hours. The mixture was diluted with a mixed solvent (150 mL) of dichloromethane:methanol = 1:1, filtered, and the filtrate was concentrated under reduced pressure. The residual compound was separated and purified by silica gel column chromatography (mobile phase: methanol:dichloromethane = 0%-15%) to obtain 175 mg of the title compound as a yellow solid, with a yield of 80.0%.

[0693] LC-MS: m / z = 266.5 [M+H] + .

[0694] Step 5: Preparation of 2-(hydroxymethyl)dithiophene[3,2-b:3',2'-d]pyridin-5(4H)-one (33e).

[0695] At 0°C, lithium aluminum hydride (2.5M in THF, 840μL, 2.11mmol) was added dropwise to a solution of methyl 5-oxo-4,5-dihydrodiino[3,2-b:3',2'-d]pyridine-2-carboxylate (175mg, 660μmol) in tetrahydrofuran (8.0mL), and stirred at 0°C for 2 hours. The reaction solution was quenched by adding methanol (20mL), filtered, and the filtrate was concentrated under reduced pressure. The residual compound was separated and purified by silica gel column chromatography (mobile phase: methanol: dichloromethane = 0%-15%) to obtain 216mg of the title compound as a white solid.

[0696] LC-MS: m / z = 238.3 [M+H] + .

[0697] Step 6: Preparation of 2-(chloromethyl)dithienyl[3,2-b:3',2'-d]pyridin-5(4H)-one (33f).

[0698] At room temperature, N,N-dimethylformamide (35 μL, 449 μmol) and thionyl chloride (391 μL, 5.39 mmol) were added to a solution of 2-(hydroxymethyl)dithiophene[3,2-b:3',2'-d]pyridin-5(4H)-one (213 mg, 898 μmol) in dichloromethane (8.0 mL), and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: dichloromethane:methanol=5%-15%) to obtain 133 mg of the title compound as a white solid, with a yield of 57.9%.

[0699] LC-MS: m / z = 256.2 [M+H] + .

[0700] Step 7: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydrodieno[3,2-b:3',2'-d]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (33).

[0701] At room temperature, N,N-diisopropylethylamine (199 μL, 1.14 mmol) and sodium iodide (68.6 mg, 458 μmol) were added to a solution of 2-(chloromethyl)dithienyl[3,2-b:3',2'-d]pyridin-5(4H)-one (133 mg, 229 μmol) and N-methyl-5-(piperazin-1-yl)picolinamide (51.0 mg, 229 μmol) in acetonitrile (6.0 mL), and the mixture was stirred at room temperature for 2 hours. The mixture was concentrated under reduced pressure, and the residual compound was separated and purified by silica gel column chromatography (mobile phase: methanol: dichloromethane = 5%-15%) to obtain 42.0 mg of the title compound as a white solid. Yield: 41.8%.

[0702] LC-MS: m / z=439.85[M+H] + .

[0703] 1 H NMR (400MHz, DMSO-d6) δ12.17(s,1H),8.40(t,J=4.8Hz,1H),8.27(d,J=2.8Hz,1H),8.15(d,J=5.2Hz,1H),7.83(d,J=8.8Hz,1H),7.56( d,J=5.2Hz,1H),7.39(dd,J=8.8,2.8Hz,1H),7.01(s,1H),3.85(s,2H),3.37–3.35(m,4H),2.78(d,J=4.8Hz,3H),2.64(t,J=5.2Hz,4H).

[0704] Example 34: Preparation of N-methyl-5-(4-((7-methyl-5-oxo-4,5-dihydrodieno[3,2-b:3',2'-d]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (34).

[0705]

[0706] Referring to the preparation method of Example 33, 3-bromothiophene-2-carboxylic acid methyl ester in step 3 was replaced by 3-bromo-5-methylthiophene-2-carboxylic acid methyl ester.

[0707] LC-MS: m / z=453.85[M+H] + .

[0708] 1 H NMR (400MHz, DMSO-d6) δ12.10(s,1H),8.41(q,J=4.8Hz,1H),8.27(d,J=2.8Hz,1H),7.83(d,J=8.8Hz,1H),7.39(dd,J=8.8,2.8Hz, 1H),7.29–7.24(m,1H),6.99(s,1H),3.84(s,2H),3.36(s,4H),2.77(d,J=4.8Hz,3H),2.63(d,J=3.6Hz,4H),2.60(d,J=1.2Hz,3H).

[0709] Example 35: Preparation of N-methyl-5-(4-((7-methyl-5-oxo-5,7-dihydro-4H-pyrazolo[4,3-d]thieno[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (35).

[0710]

[0711] Referring to the preparation method of Example 33, the 3-(tributyltinyl)thiophene-2-carboxylic acid methyl ester in step 3 was replaced with 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole-3-carboxylic acid methyl ester.

[0712] LC-MS: m / z=438[M+H] + .

[0713] 1 H NMR (400MHz, DMSO-d6) δ11.82(s,1H),8.82–8.38(m,2H),8.33(d,J=2.9Hz,1H),7.88(d,J=8.7Hz,1H),7.47(dd,J= 8.8, 2.9Hz, 1H), 7.23 (s, 1H), 4.67 (d, J = 15.5Hz, 2H), 4.11 (s, 3H), 3.52 (s, 4H), 3.18 (s, 4H), 2.78 (d, J = 4.8Hz, 3H).

[0714] Example 36: Preparation of N-methyl-5-(4-(5-oxo-4,5-dihydropyrrolo[1,2-a]quinazolin-2-yl)methyl)piperazine-1-piperidinecarboxamide (36).

[0715]

[0716] Step 1: Preparation of 2-formylsuccinonitrile potassium salt (mixture of E-form / Z-form) (36a)

[0717] At 0°C, dissolve succinonitrile (5.00 g, 62.4 mmol) and ethyl formate (6.03 mL, 74.9 mmol) in toluene (30 mL) and tert-butanol (10 mL), add a solution of potassium tert-butoxide (7.15 g, 63.6 mol) in tert-butanol (50 mL), and stir at room temperature for 2 hours. Filter and dry the filter cake to obtain 6.15 g of the title compound as a light brown solid, with a yield of 67.4%.

[0718] Step 2: 2-(2,3-Dicyanopropen-1-ylamino)benzoic acid methyl ester (36b) (mixture of E-form / Z-form)

[0719] At room temperature, 2-formylsuccinonitrile potassium salt (2.00 g, 13.2 mmol) (mixture of E type / Z type) and methyl anthranilate (1.93 g, 13.2 mmol) were dissolved in acetic acid (6 mL) and water (6 mL), and reacted at 100°C for 10 minutes. The mixture was cooled to room temperature naturally, filtered, and the filter cake was dried to obtain 2.24 g of the title compound as a light brown solid, with a yield of 75.5%.

[0720] Step 3: Preparation of 5-oxo-4,5-dihydropyrrolo[1,2-a]quinazoline-2-carbonitrile (36c)

[0721] At 0°C, methyl 2-(2,3-dicyanopropen-1-ylamino)benzoate (2.00 g, 8.29 mmol) (mixture of E-type / Z-type) was dissolved in ethanol (15 mL), and a solution of sodium ethoxide (1.41 g, 20.7 mmol) in ethanol (25 mL) was added dropwise, and the mixture was reacted at room temperature for 2 hours. 1M hydrochloric acid (20 mL) was added, filtered, and the filter cake was dried to obtain 1.42 g of the title compound as a white solid, with a yield of 81.9%.

[0722] LC-MS: m / z = 210.2 [M+H] + .

[0723] Step 4: Preparation of 5-oxo-4,5-dihydropyrrolo[1,2-a]quinazoline-2-carbaldehyde (36d)

[0724] At 0°C, 5-oxo-4,5-dihydropyrrolo[1,2-a]quinazoline-2-carbonitrile (1.30 g, 6.21 mmol) was dissolved in ultra-dry dichloromethane (150 mL), and diisobutylaluminum hydride (8.28 mL, 12.4 mmol, 1.5 M in toluene) was slowly added dropwise. Stirring was maintained at 0°C for 1 hour, and stirring was continued at room temperature for 3 hours. 1 M hydrochloric acid was added, and stirring was continued at room temperature for 10 minutes. A mixed solvent of dichloromethane: methanol (4:1) was added to fully dissolve the solid, and the liquids were separated. The aqueous layer was extracted three times with a mixed solvent (100 mL x 3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 50:1-4:1) to obtain 427 mg of the title compound as a yellow solid, with a yield of 32.1%.

[0725] LC-MS: m / z = 213.2 [M+H] + .

[0726] Step 5: Preparation of N-methyl-5-(4-(5-oxo-4,5-dihydropyrrolo[1,2-a]quinazolin-2-yl)methyl)piperazine-1-piperidinecarboxamide (36)

[0727] At room temperature, N-methyl-5-piperazine-1-piperidinecarboxamide (568 mg, 2.22 mmol) and triethylamine (224 mg, 2.22 mmol) were dissolved in 1,2-dichloroethane (5 mL) and methanol (5 mL), stirred for 30 minutes, added to a solution of 5-oxo-4,5-dihydropyrrolo[1,2-a]quinazoline-2-carbaldehyde (427 mg, 2.01 mmol) in acetic acid (3 mL), stirred at room temperature for 30 minutes, added sodium cyanoborohydride (253 mg, 4.03 mol), and stirred at 70°C for 16 hours. The reaction solution was filtered through diatomaceous earth, the filtrate was collected and concentrated under reduced pressure, and the residue was separated by high-pressure preparative liquid separation (chromatographic column model: Daisogei 30mm*250mm, C18, 10um100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid) to obtain 143 mg of the title compound as a white solid, with a yield of 17.1%.

[0728] LC-MS: m / z=417.1[M+H] + .

[0729] 1 H NMR (400MHz, DMSO-d6) δ11.91(s,1H),8.45(q,J=4.8Hz,1H),8.34(d,J=2.9Hz,1H),8.14(d,J=7.9Hz,1H),7.97(d,J=8.3Hz,1H),7.90–7.82(m,2H) ,7.72(s,1H),7.49–7.43(m,2H),5.81(s,1H),4.32(d,J=3.5Hz,2H),4.1 0(s,1H),3.54(d,J=9.8Hz,2H),3.23–3.11(m,1H),2.79(d,J=4.7Hz,2H).

[0730] Example 37: Preparation of N-methyl-5-(4-(8-fluoro-5-oxo-4,5-dihydropyrrolo[1,2-a]quinazolin-2-yl)methyl)piperazine-1-piperidinecarboxamide (37).

[0731]

[0732] Referring to the preparation method of Example 36, the methyl o-aminobenzoate in step 2 was replaced by methyl 2-amino-4-fluorobenzoate.

[0733] LC-MS: m / z 435[M+H] + .

[0734] 1H NMR (400MHz, DMSO-d6) δ11.73(s,1H),8.39(q,J=4.9Hz,1H),8.26(d,J=2.9Hz,1H), 8.13(dd,J=8.8,6.3Hz,1H),7.92(dd,J=10.4,2.4Hz,1H),7.82(d,J=8.8Hz,1H),7.5 4–7.47(m,1H),7.39(dd,J=8.9,2.9Hz,1H),7.20(td,J=8.6,2.4Hz,1H),5.59(d,J= 1.7Hz,1H),3.51(d,J=9.7Hz,3H),3.35(s,26H),2.77(d,J=4.8Hz,3H),2.61(s,5H).

[0735] Example 38: Preparation of N-methyl-5-(4-((5-oxo-4,5-dihydropyrazolo[1,5-a]quinazolin-2-yl)methyl)piperazin-1-yl)picolinamide (38).

[0736]

[0737] Referring to the preparation method of Example 31, the 2-chloronicotinic acid chloride in step 1 was replaced by 2-fluorobenzoyl chloride.

[0738] LC-MS: m / z=417.95[M+H] + .

[0739] 1 H NMR (400MHz, DMSO-d6) δ12.16(s,1H),8.39(q,J=4.8Hz,1H),8.26(d,J=2.8Hz,1H ),8.13(dd,J=8.0,1.6Hz,1H),8.04(d,J=8.4Hz,1H),7.86(td,J=8.4,8.0,1.6Hz, 1H),7.82(d,J=8.8Hz,1H),7.47(t,J=7.6Hz,1H),7.38(dd,J=8.8,2.8Hz,1H),5. 87(s,1H),3.62(s,2H),3.35(s,4H),2.77(d,J=4.8Hz,3H),2.61(t,J=5.2Hz,4H).

[0740] Example 39: Preparation of 5-(4-((8-fluoro-5-oxo-4,5-dihydropyrazolo[1,5-a]quinazolin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (39).

[0741]

[0742] Referring to the preparation method of Example 31, the 2-chloronicotinic acid chloride in step 1 was replaced by 2-chloro-4-fluorobenzoyl chloride.

[0743] LC-MS: m / z=435.85[M+H] + .

[0744] 1 H NMR (400MHz, DMSO-d6) δ12.26(s,1H),8.40(q,J=4.8Hz,1H),8.27(d,J=2.8Hz,1H),8.19(dd,J=8.8,6.0Hz,1H),7.83(d,J=8.8Hz,1H),7.74(dd,J=9.6 ,2.4Hz,1H),7.40(dd,J=8.8,2.8Hz,1H),7.33(td,J=8.8,2.8Hz,1H),5.95 (s,1H),3.69(s,2H),3.40(s,4H),2.77(d,J=4.8Hz,3H),2.75–2.51(m,4H).

[0745] Example 40: Preparation of N-methyl-5-(4-((3-methyl-5-oxo-4,5-dihydrothieno[3,2-c]isoquinolin-2-yl)methyl)piperazin-1-yl)picolinamide (40).

[0746]

[0747] Step 1: Preparation of methyl 5-(2-(methoxycarbonyl)phenyl)-3-methylthiophene-2-carboxylate (40a).

[0748] At room temperature and under nitrogen atmosphere, methyl 3-methylthiophene-2-carboxylate (7.00 g, 44.9 mmol) was dissolved in 1,4-dioxane (60.0 mL) and dimethyl sulfoxide (11.0 mL), and methyl 2-iodobenzoate (11.8 g, 44.9 mmol), palladium acetate (504 mg, 2.25 mmol), tricyclohexylphosphine (1.26 g, 4.49 mmol) and silver carbonate (24.8 g, 89.8 mmol) were added and reacted at 110 °C for 16 hours. The mixture was filtered and concentrated under reduced pressure. The residue was diluted with water (60.0 mL) and extracted with ethyl acetate (60.0 mL x 3). The organic phase was washed with saturated brine (60.0 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1-9:1) to obtain 6.02 g of the title compound as a yellow solid. The yield was 46.2%.

[0749] LC-MS: m / z = 291.1 [M+H] + .

[0750] Step 2: Preparation of methyl 5-(2-(methoxycarbonyl)phenyl)-3-methyl-4-nitrothiophene-2-carboxylate (40b).

[0751] At 0°C, under nitrogen atmosphere, methyl 5-(2-(methoxycarbonyl)phenyl)-3-methylthiophene-2-carboxylate (6.02 g, 20.7 mmol) was dissolved in acetic anhydride (60.0 mL), and concentrated nitric acid (2.49 g, 24.9 mmol) was added dropwise, and the mixture was reacted at room temperature for 4 hours. Water (50.0 mL) was added for dilution, and the pH was adjusted to 7-8 with saturated sodium bicarbonate solution, and the mixture was extracted with ethyl acetate (100.0 mL x 3). The organic phase was washed with saturated brine (100.0 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1-9:1) to obtain 2.96 g of the title compound as a yellow solid with a yield of 42.6%.

[0752] LC-MS: m / z=336.1[M+H] + .

[0753] Step 3: Preparation of methyl 3-methyl-5-oxo-4,5-dihydrothieno[3,2-c]isoquinoline-2-carboxylate (40c).

[0754] At room temperature and under nitrogen atmosphere, methyl 5-(2-(methoxycarbonyl)phenyl)-3-methyl-4-nitrothiophene-2-carboxylate (2.96 g, 8.83 mmol) was dissolved in ethanol (30.0 mL), iron powder (1.97 g, 35.3 mmol) and 6M aqueous hydrochloric acid solution (3.0 mL, 18.0 mmol) were added, and the mixture was reacted at 80°C for 16 hours. The mixture was filtered, the filter cake was washed with methanol (50 mL), the filtrate was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: petroleum ether / ethyl acetate = 100:1-3:1) to obtain 97.2 mg of the title compound as a yellow solid, with a yield of 4.0%.

[0755] LC-MS: m / z 273.9[M+H] + .

[0756] Step 4: Preparation of 2-hydroxymethyl-3-methylthieno[3,2-c]isoquinolin-5(4H)-one (40d).

[0757] At room temperature and under nitrogen atmosphere, methyl 3-methyl-5-oxo-4,5-dihydrothieno[3,2-c]isoquinoline-2-carboxylate (97.2 mg, 0.356 mmol) was dissolved in tetrahydrofuran (2.0 mL), and lithium aluminum hydride (0.29 mL, 0.712 mmol, 2.5 M in THF) was added dropwise, and the mixture was reacted at room temperature for 2 hours. Sodium sulfate decahydrate was added to quench the reaction, and the mixture was filtered. The filter cake was washed with methanol (10.0 mL), and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (mobile phase: dichloromethane / methanol = 100:1-19:1) to obtain 72.1 mg of the title compound as a yellow solid, with a yield of 82.5%.

[0758] LC-MS: m / z = 245.9 [M+H] + .

[0759] Step 5: Preparation of 3-methyl-5-oxo-4,5-dihydrothieno[3,2-c]isoquinoline-2-carbaldehyde (40e).

[0760] At room temperature under nitrogen atmosphere, 2-hydroxymethyl-3-methylthieno[3,2-c]isoquinolin-5(4H)-one

[0761] (72.1 mg, 0.294 mmol) was dissolved in dichloromethane (10.0 mL), and Dess-Martin periodinane (249 mg, 0.588 mmol) was added, and the mixture was reacted at 40°C for 16 hours. The mixture was concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (mobile phase: dichloromethane / methanol = 100:1-19:1) to obtain 62.3 mg of the title compound as a yellow solid, with a yield of 87.1%.

[0762] LC-MS: m / z = 243.9 [M+H] + .

[0763] Step 6: Preparation of N-methyl-5-(4-((3-methyl-5-oxo-4,5-dihydrothieno[3,2-c]isoquinolin-2-yl)methyl)piperazin-1-yl)picolinamide (40).

[0764] At room temperature and under nitrogen atmosphere, N-methyl-5-piperazin-1-ylpicolinamide hydrochloride (72.3 mg, 0.282 mmol) was dissolved in 1,2-dichloroethane (3.0 mL) and methanol (3.0 mL), and triethylamine (28.5 mg, 0.282 mmol) was added. After reacting at room temperature for 0.5 hour, acetic acid (0.1 mL) and 3-methyl-5-oxo-4,5-dihydrothieno[3,2-c]isoquinoline-2-carbaldehyde (62.3 mg, 0.256 mmol) were added. After reacting at room temperature for 0.5 hour, sodium cyanoborohydride (32.2 mg, 0.512 mmol) was added, and the mixture was reacted at 60°C for 16 hours. The residue was concentrated under reduced pressure and separated by high pressure preparative liquid chromatography (chromatographic column model: Daisogei 30 mm*250 mm, C18, 10 um 100A, mobile phase: acetonitrile / water, gradient: 10%-50%, 0.05% formic acid) to obtain 23.0 mg of the title compound as a yellow solid in a yield of 11.5%.

[0765] LC-MS: m / z=448.0[M+H] + .

[0766] 1 H NMR (400MHz, DMSO-d6) δ11.86(s,1H),8.45(q,J=4.7Hz,1H),8.37–8.26(m,2H),7.88(d,J=8.8Hz,1H),7.84–7.76(m,2H),7.58 (t,J=7.1Hz,1H),7.47(dd,J=8.9,2.9Hz,1H),4.69(s,2H),3.54(s,0H),3.34–3.14(m,3H),2.78(d,J=4.8Hz,3H),2.39(s,3H).

[0767] Example 41: Preparation of N-methyl-5-(4-((5-oxo-4,5,6,7-tetrahydrofuro[2,3-d]thieno[3,2-b]pyridin-2-yl)methyl)piperazin-1-yl)picolinamide (41).

[0768]

[0769] Referring to the preparation method of Example 17, the 2-oxocyclopentane-1-carboxylic acid methyl ester in step 1 was replaced by 2-oxotetrahydrofuran-3-carboxylic acid methyl ester.

[0770] LC-MS: m / z = 424.9 [M+H] + .

[0771] 1H NMR (400MHz, DMSO-d6) δ12.01(s,1H),8.45(d,J=4.9Hz,1H),8.33(d,J=2.9Hz,1H),7.88(d,J=8.7Hz,1H),7.47(dd,J=8.8,2.9H z,1H),7.26(s,1H),4.79(t,J=9.1Hz,2H),4.66(s,4H),4.03(s,1H),3.25(s,6H),3.03(t,J=9.1Hz,2H),2.79(d,J=4.8Hz,3H).

[0772] Example 42: Preparation of N-methyl-5-(4-((6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[2,3-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (42).

[0773]

[0774] Referring to the preparation method of Example 22, 5-chloro-4-nitrothiophene-2-carboxylic acid methyl ester in step 1 was replaced by 4-bromo-5-nitrothiophene-2-carboxylic acid methyl ester.

[0775] LC-MS: m / z=437.0[M+H] + .

[0776] 1 H NMR (400MHz, DMSO-d6) δ11.18(s,1H),8.43(d,J=5.2Hz,1H),8.28(d,J=2.9Hz,1H) ,7.84(d,J=8.7Hz,1H),6.44(d,J=2.7Hz,1H),2.78(d,J=4.8Hz,3H),2.48(s,2H).

[0777] Example 43: Preparation of 5-(4-((6-cyano-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (43).

[0778]

[0779] Referring to the preparation method of Example 22, 3-methylpyrrole-2-carboxylic acid methyl ester in step 1 was replaced by 3-cyanopyrrole-2-carboxylic acid methyl ester.

[0780] LC-MS: m / z=448.5[M+H] + .

[0781] 1 H NMR (400MHz, DMSO-d6) δ12.22(s,1H),8.44(q,J=4.3Hz,1H),8.33(d,J=2.5Hz,1H),7.97(d,J=2.6Hz,1H),7.88(d,J=8.8Hz,1H),7 .47(dd,J=8.8,2.6Hz,1H),7.23(d,J=2.9Hz,1H),7.21–7.10(m,1H),4.57(s,2H),3.94(s,4H),3.25(s,4H),2.78(d,J=4.8Hz,3H).

[0782] Example 44: Preparation of 6-fluoro-N-methyl-5-(4-((6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (44)

[0783]

[0784] Referring to the preparation method of Example 22, the N-methyl-5-piperazin-1-ylpicolinamide hydrochloride in step 5 was replaced by 4-fluoro-N-methyl-5-piperazin-1-ylpicolinamide.

[0785] LC-MS: m / z = 458.2 [M+H] + .

[0786] 1 H NMR (400MHz, DMSO-d6) δ11.40–11.25(m,1H),8.44(q,J=4.5Hz,1H),7.88(d,J=7.9Hz,1H),7.75–7.62(m,1H),7.62– 7.51(m,1H),7.16–7.04(m,1H),6.49(d,J=2.2Hz,1H),4.63(s,2H),3.72(s,4H),3.18(s,4H),2.77(d,J=4.7Hz,3H).

[0787] Example 45: Preparation of N,6-dimethyl-5-(4-((6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (45)

[0788]

[0789] Referring to the preparation method of Example 22, the N-methyl-5-piperazin-1-ylpicolinamide hydrochloride in step 5 was replaced by 4-methyl-N-methyl-5-piperazin-1-ylpicolinamide.

[0790] LC-MS: m / z=458[M+H] + .

[0791] 1 H NMR (400MHz, DMSO-d6) δ11.11(s,1H),8.41(q,J=4.5Hz,1H),7.79(d,J=8.2Hz,1H),7.49(d,J=8.3Hz,1H),7.44(d,J=2.5Hz,1H),6.7 7(s,1H),6.42(d,J=2.5Hz,1H),3.78(s,2H),3.30(s,3H),2.96(s,4H),2.80(d,J=4.8Hz,3H),2.66(s,4H),2.49(s,3H),2.48(s,2H).

[0792] Example 46: Preparation of 5-(4-((6-chloro-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)-N,6-dimethylpicolinamide (46)

[0793]

[0794] Referring to the preparation method of Example 28, N-methyl-5-piperazin-1-ylpicolinamide hydrochloride was replaced by 4-methyl-N-methyl-5-piperazin-1-ylpicolinamide.

[0795] LC-MS: m / z=471.2[M+H] + .

[0796] 1 H NMR(400MHz,Chloroform-d)δ9.52(s,1H),7.99(d,J=8.2Hz,1H),7.95(d,J=5.0Hz,1H),7.35(d,J=8.3Hz,1H),7.10(d,J= 2.8Hz,1H),6.76(s,1H),6.59(d,J=2.8Hz,1H),3.78(s,2H),3.01(d,J=5.0Hz,4H),2.73(s,4H),2.51(s,3H),1.25(s,3H).

[0797] Example 47: Preparation of 5-(4-((6-chloro-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)-6-fluoro-N-methylpicolinamide (47)

[0798]

[0799] Referring to the preparation method of Example 28, N-methyl-5-piperazin-1-ylpicolinamide hydrochloride was replaced with 4-fluoro-N-methyl-5-piperazin-1-ylpicolinamide.

[0800] LC-MS: m / z=475[M+H] + .

[0801] 1 H NMR(400MHz,Chloroform-d)δ10.37(s,1H),8.00(dd,J=8.0,1.3Hz,1H),7.50(q,J=5.4,4.9Hz,1H),7.31(dd,J=10.2,8.0Hz,1H),7.10 (d,J=2.9Hz,1H),6.82(s,1H),6.59(d,J=2.9Hz,1H),3.79(s,2H),3.26(t,J=4.7Hz,4H),2.99(d,J=5.1Hz,3H),2.75(t,J=4.8Hz,4H).

[0802] Example 48: Preparation of 1'-((6-chloro-5-oxo-4,5-dihydropyrrolo[1,2-e]thieno[3,2-e]pyrazin-2-yl)methyl)-N-methyl-1',2',3',6'-tetrahydro-[3,4-bipyridine]-6-carboxamide (48)

[0803]

[0804] Referring to the preparation method of Example 28, N-methyl-5-piperazin-1-ylpicolinamide hydrochloride was replaced with N-methyl-1',2',3',6'-tetrahydro-[3,4'-bipyridine]-6-carboxamide.

[0805] LC-MS: m / z=454[M+H] + .

[0806] 1H NMR (400MHz, DMSO-d6) δ11.67(d,J=12.8Hz,1H),10.30(s,1H),8.85–8.61(m,1H),8.16–7.95(m,2H),7.80(s,1H), 7.19(s,1H),6.74(dd,J=9.6,3.0Hz,1H),6.43(d,J=3.9Hz,1H),3.91(s,2H),3.29(s,2H),3.03–2.74(m,4H),1.43–

[0807] 1.02(m,3H).

[0808] Example 49: Preparation of 5-(4-((6-ethyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)-N-methylpicolinamide (49)

[0809]

[0810] Referring to the preparation method of Example 22, 3-methylpyrrole-2-carboxylic acid methyl ester in step 1 was replaced by 3-ethylpyrrole-2-carboxylic acid methyl ester.

[0811] LC-MS: m / z=451[M+H] + .

[0812] 1 H NMR (400MHz, DMSO-d6) δ11.35(s,1H),8.46(t,J=4.8Hz,1H),8.34(d,J=2.9H z,1H),7.89(d,J=8.8Hz,1H),7.60(d,J=2.7Hz,1H),7.48(dd,J=8.8,2.9Hz, 1H),7.12(s,1H),6.56(d,J=2.7Hz,1H),4.09(s,2H),3.53(s,4H),3.17(s,4 H), 2.96 (q, J = 7.5Hz, 2H), 2.79 (d, J = 4.8Hz, 3H), 1.20 (q, J = 8.0, 7.6Hz, 3H).

[0813] Example 50: Preparation of N-(methyl-d3)-5-(4-((6-methyl-5-oxo-4,5-dihydropyrrolo[1,2-a]thieno[3,2-e]pyrazin-2-yl)methyl)piperazin-1-yl)picolinamide (50)

[0814]

[0815] Referring to the preparation method of Example 22, the N-methyl-5-piperazine-1-ylpicolinamide hydrochloride in step 1 was replaced by N-(methyl-d3)-5-(piperazine-1-yl)picolinamide.

[0816] LC-MS: m / z=439.90[M+H] + .

[0817] 1 H NMR (400MHz, DMSO-d6) δ11.36(s,1H),8.44(s,1H),8.34(d,J=2.8Hz,1H),7.88(d,J=8.8Hz,1H),7.58(d,J=2.8Hz,1H),7.48(dd, J=8.8,2.8Hz,1H),7.11(s,1H),6.49(d,J=2.8Hz,1H),4.64(s,2H),4.43(s,4H),4.09(s,2H),3.50(s,2H),2.49(d,J=1.2Hz,3H).

[0818] Biological evaluation

[0819] Experimental Example 1: PARP1 capture experiment

[0820] Experimental materials: test compound, PARP1 (BPS, 80501), NAD (Sigma, 10127965001), DSB DNA probe-1 (Kanglong Chemical), Mab anti GST-Tb cryptate (cisbio, 61GSTTLA)

[0821] Experimental steps:

[0822] (I) Reagent preparation

[0823] Prepare assay buffer: 10 mM potassium phosphate (pH 7.9), 50 mM NaCl, 1 mM EDTA, 0.05% Brij-35, 1 mM DTT.

[0824] (II) Experimental methods

[0825] The buffer solution was prepared by the above method, the compound was dissolved in DMSO and diluted in a gradient manner, the compound was diluted to the test concentration using the experimental buffer, and the solution was shaken on a shaker for 15 min.

[0826] PARP1 enzyme was diluted 4X using assay buffer and GST-Tb was added simultaneously.

[0827] Add the prepared enzyme solution to the assay plate, 4 μL per well.

[0828] Dilute PARP1 probe 1 to 4X using assay buffer and add 4 μL per well to the assay plate.

[0829] The compounds to be tested were added to the experimental plate, 4 μL per well, and incubated in a room temperature incubator for 1 hour.

[0830] Dilute NAD to 4X using assay buffer, add 4 μL per well to the assay plate, and incubate in a room temperature incubator for 10 minutes.

[0831] The final values ​​were read by Envision (2105) (Ex: 340 nm, Em 665 nm and 615 nm).

[0832] Data Analysis:

[0833] The % inhibition rate was calculated as follows:

[0834] Suppression%=(Signal cmpd-Signal Ave_PC) / (Signal Ave_VC-Signal Ave_PC)

[0835] ×100

[0836] Signal cmpd: Fluorescence signal values ​​of compounds at different concentrations

[0837] SignalAve_PC: The average value of the maximum fluorescence signal of the system

[0838] SignalAve_VC: Average value of negative control fluorescence signal value

[0839] Calculation IC 50 , draw the compound effect dose curve:

[0840] IC was calculated by fitting the percent inhibition values ​​and logarithms of compound concentrations to a nonlinear regression (dose response - variable slope) using Graphpad 5.0. 50 .

[0841] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0842] X: logarithmic value of inhibitor concentration; Y: percentage of inhibition rate

[0843] Table 1 provides the in vitro activities of the compounds of the present invention on PARP1 capture (IC 50 ).

[0844] In Table 1, the PARP1 capture in vitro activity values ​​of the compounds are shown as follows: A refers to IC 50 <10nM; B refers to 10nM <IC 50 <100nM; C refers to 100nM <IC 50 <1000nM; D refers to IC 50 >1000nM.

[0845] Table 1 Activity of the compounds of the present invention on PARP1 capture

[0846]

[0847]

[0848] Results: The compounds of the present invention have good capture activity on PARP1-DNA complex.

[0849] Experimental Example 2: PARP2 capture experiment

[0850] Experimental materials: test compounds, PARP2 (BPS, 80502), NAD (Sigma, 10127965001), DSB DNA probe-2 (Pharmaron Chemicals), Mab anti GST-Tbcryptate (cisbio, 61GSTTLA).

[0851] Experimental steps:

[0852] (I) Reagent preparation

[0853] Prepare assay buffer: 10 mM potassium phosphate (pH 7.9), 50 mM NaCl, 1 mM EDTA, 0.05% Brij-35, 1 mM DTT.

[0854] (II) Experimental methods:

[0855] The reagents were prepared by the above method. The compounds were dissolved in DMSO and serially diluted, and the compounds were diluted to the test concentration using the assay buffer and shaken on a shaker for 15 min.

[0856] PARP2 enzyme was diluted 4X using assay buffer and GST-Tb was added simultaneously.

[0857] Add the prepared enzyme solution to the assay plate, 4 μL per well.

[0858] Dilute PARP2 probe-2 to 4X using assay buffer and add 4 μL per well to the assay plate.

[0859] The compounds to be tested were added to the experimental plate, 4 μL per well, and incubated in a room temperature incubator for 1 hour.

[0860] Dilute NAD to 4X using assay buffer, add 4 μL per well to the assay plate, and incubate in a room temperature incubator for 10 minutes.

[0861] The final values ​​were read by Envision (2105) (Ex: 340 nm, Em 665 nm and 615 nm).

[0862] Data Analysis:

[0863] The % inhibition rate was calculated as follows:

[0864] Suppression%=(Signal cmpd-Signal Ave_PC) / (Signal Ave_VC-Signal Ave_PC)

[0865] ×100

[0866] Signal cmpd: Fluorescence signal values ​​of compounds at different concentrations

[0867] SignalAve_PC: The average value of the maximum fluorescence signal of the system

[0868] SignalAve_VC: Average value of negative control fluorescence signal value

[0869] Calculation IC 50 , draw the compound effect dose curve:

[0870] IC was calculated by fitting the percent inhibition values ​​and logarithms of compound concentrations to a nonlinear regression (dose response - variable slope) using Graphpad 5.0. 50 .

[0871] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0872] X: Logarithmic value of inhibitor concentration; Y: Percentage of inhibition rate

[0873] Table 2 provides the in vitro activities of the compounds of the invention on PARP2 capture (IC 50 ).

[0874] In Table 2, the PARP2 capture in vitro activity values ​​of the compounds are shown as follows: A refers to IC 50 <10nM; B refers to 10nM <IC 50 <100nM; C refers to 100nM <IC50 <1000nM; D refers to IC 50 >1000nM.

[0875] Table 2 Activity of the compounds of the present application on PARP2 capture

[0876] Example <![CDATA[PARP2 Capture IC 50 > Example <![CDATA[PARP2 Capture IC 50 > 3 D 25 D 6 D 28 C 7 D 34 C 9 D 36 D 11 D 44 D 17 D 45 D 18 D 46 C 21 D 47 C 22 D 48 C 23 C 49 D 24 C 50 D

[0877] Results: The compounds of the present invention have low capture activity on PARP2-DNA complex.

[0878] Experimental Example 3: Antiproliferation experiment of DLD-1 wild type (WT) and DLD-1BRCA2(- / -) cells

[0879] Experimental materials: test compounds, RPMI1640 medium (Corning, 10-040-CV), fetal bovine serum (AusGeneX, FBS500-S), penicillin / streptomycin antibiotics (Gibco, 15140-122), CelltiterGlo analysis kit (CTG) (Promega, G7573), DLD-1WT (Punosai, CL-0074) and DLD-1BRCA2(- / -) (Horizon, HD 105-007) cell lines.

[0880] The experimental method is as follows:

[0881] On the first day, Echo transferred 40 nl of the test compound (10 mM, dissolved in DMSO) into a 384-well plate, seeded DLD-1WT and DLD-1BRCA2(- / -) cells in a white 384-well plate, added 40 ul of cell suspension to each well, containing 600 DLD-1WT or DLD-1BRCA2(- / -) cells, and cultured the cell plate in a 37°C, 5% carbon dioxide incubator for 7 days.

[0882] Then, Promega CellTiter-Glo assay was performed. The cell plate was taken out and equilibrated at room temperature for 30 minutes. 20 μL CTG was added to each well, mixed by vortexing, and incubated at room temperature for 30 minutes. The luminescence was read by Envision (2105) multi-label analyzer.

[0883] Data Analysis:

[0884] The inhibition rate was calculated as follows:

[0885] Inhibition% = (Signalcmpd-SignalAve_PC) / (SignalAve_VC-SignalAve_PC)×100

[0886] Signal cmpd: Chemiluminescence values ​​of compounds at different concentrations

[0887] SignalAve_PC: The average value of the maximum chemiluminescence value of the system

[0888] SignalAve_VC: Average value of negative control chemiluminescence value

[0889] Calculation IC 50 , draw the compound effect dose curve:

[0890] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0891] X: Logarithmic value of compound concentration; Y: Percentage of inhibition rate

[0892] Table 3 provides the inhibitory activity of the compounds of the present invention on the proliferation of DLD-1WT and DLD-1BRCA2(- / -) cells.

[0893] In Table 3, the inhibitory activity values ​​of the compounds on the proliferation of DLD-1WT and DLD-1BRCA2(- / -) cells: A refers to IC 50 <100nM; B refers to 100nM <IC 50 <1000nM; C refers to 1000nM <IC 50 <10000nM; D refers to IC 50 >10000nM; “—” means not tested.

[0894] Table 3 Inhibitory activity of the compounds of the present invention on proliferation of DLD-1WT and DLD-1BRCA2(- / -) cells

[0895] Example DLD-1BRCA2(- / -)(nM) DLD-1WT(nM) 3 D — 4 A D 6 A D 17 A D 18 A D 21 A D 22 A D 23 B — 28 A D 34 A D 36 B — 44 A D 45 A D 46 A D 47 A D 49 A D 50 A D

[0896] Results: The compounds of the present invention have good antiproliferative activity against BRCA2 mutated DLD-1 cells, and almost no inhibitory activity against wild-type DLD-1 cells, indicating that the compounds of the present invention have good cell selectivity.

[0897] Test Example 4: Determination of proliferation inhibition activity of triple-negative breast cancer MDA-MB-436 cells

[0898] Experimental materials: MDA-MB-436 cells (Wuhan Prosai Life Science Technology Co., Ltd., CL-0383A), Leibovitz's L-15 cell culture medium (Wuhan Prosai Life Science Technology Co., Ltd., CM-0383A), PBS (Gibco, 8121763), CellTiter Glo assay kit (Promega, G7572), 384-well plate (Thermo, 164610), and microplate reader (Biotek, Cytation 3).

[0899] Experimental method: MDA-MB-436 cells were cultured in a special Leibovitz's L-15 medium. After thawing, the cells were passaged 1-3 times. The cells were collected and centrifuged at 1000 rpm for 5 min to prepare a cell suspension. The cells were counted and the cell density was adjusted to 1.33×10 4 / mL. Add the cell suspension to a 384-well plate, 45uL per well, i.e. 600 live cells / well. Use DMSO and culture medium to prepare compound solutions of different concentrations, and set up solvent control wells and blank control wells. Take 5μL of compound solution and add it to 45μL cells in a 384-well plate, 2 replicates, and the final DMSO concentration in the 50uL reaction system is 0.1%. The final concentrations of the compounds are: 10000nM, 2500nM, 625nM, 156.25nM, 39.06nM, 9.77nM, 2.44nM, 0.61nM, 0.15nM, 0nM. Place the cell culture plate in a 37°C, 5% CO2 incubator for 7 days, then take it out, balance at room temperature for 30min, add 20μL CTG to each well, shake and mix, incubate at room temperature for 10min, and read the luminescence value (Luminescence) with an enzyme reader.

[0900] The cell proliferation activity inhibition rate was calculated as follows:

[0901] Inhibition%=100-(Signalcmpd-SignalAve_PC) / (Signal Ave_VC-SignalAve_PC)×100

[0902] Signalcmpd: chemiluminescence value of each concentration of compound,

[0903] SignalAve_VC: average chemiluminescence value of solvent control,

[0904] SignalAve_PC: The average value of blank control chemiluminescence value.

[0905] The concentration and inhibition rate were fitted by nonlinear regression curve using Graphpad Prism software to obtain IC 50 value.

[0906] Y=Bottom+(Top-Bottom) / (1+10^((LogIC 50 -X)*HillSlope))

[0907] X: logarithmic value of compound concentration; Y: percentage of inhibition rate.

[0908] Table 4 provides the inhibitory activity of the compounds of the present invention on the proliferation of MDA-MB-436 cells. In Table 4, the inhibitory activity values ​​of the compounds on the proliferation of MDA-MB-436 cells: A refers to IC 50 <100nM; B refers to 100nM <IC 50 <1000nM; C refers to 1000nM <IC 50 <10000nM; D refers to IC 50 >10000nM.

[0909] Table 4 Inhibitory effect of the compounds of the present invention on the proliferation activity of MDA-MB-436 cells IC 50

[0910]

[0911]

[0912] Results: The compounds of the present invention have good antiproliferative activity against MDA-MB-436 cells.

Claims

1. A compound represented by the general formula (I) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, in: is a single bond or a double bond; A 1 and A 2 are each independently selected from N, C, O or CH; X 1 , X 2 and X 3 are each independently selected from C or N; B 1 and B 2 Each independently selected from O, N, S, NR 5 or CR 5 ; L1 is selected from a bond, a carbonyl group or a CR 6 R 7 ; L2 is selected from a bond, O, S, NR 6 or CR 6 R 7 ; R 1 and R 2 are each independently selected from hydrogen, deuterium, halogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 3-6 Cycloalkyl, C 3-5 Heterocyclic group, wherein the C 3-6 Cycloalkyl, C 3-5 The heterocyclic group is optionally substituted by 1, 2 or 3 R a Replace, or R 1 and R 2 Each does not exist; R a is selected from the group consisting of hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, mercapto, alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted with one or more substituents selected from the group consisting of deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; Or, R 1 With R 2 , A 1 , A 2 Together they form C 3-7 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 aromatic ring or 5-14 membered heteroaromatic ring, wherein the C 3-7 Hydrocarbon ring, 3-10 membered heterocyclic ring, C 6-10 The aromatic ring or 5-14 membered heteroaromatic ring is optionally replaced by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 Substitution of cycloalkyl groups; Ring B is selected from a 4-6 membered saturated monocyclic heterocycle containing 1-2 nitrogen atoms, a 5-6 membered partially unsaturated monocyclic heterocycle containing 1-2 nitrogen atoms, a 6-8 membered saturated heterocyclic bridge ring containing 1-4 nitrogen atoms, a 5-10 membered saturated heterocyclic cyclopentadiene containing 1-4 nitrogen atoms, or a 5-11 membered saturated heterocyclic spirocycle containing 1-4 nitrogen atoms; m is 0, 1, 2 or 3; Each R 3 Each independently selected from deuterium, halogen, amino, cyano, hydroxyl, C 1-6 Alkyl, C 1-6 Cycloalkyl, C 1-4 Alkoxy, halogenated C 1-4 Alkyl, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkyl, deuterated C 1-4 Alkoxy; or two R on the same atom 3 Together with the carbon atom to which it is attached, it forms C=O; Ring C is selected from a 5-6 membered monocyclic aromatic heterocycle containing 1-2 nitrogen, oxygen, or sulfur atoms, an 8-10 membered bicyclic aromatic heterocycle containing 1-2 nitrogen, oxygen, or sulfur atoms, and a 6-10 membered aromatic ring; n is 0, 1, 2 or 3; Each R 4 Each independently selected from deuterium, halogen, amino, amide, C(O)NR 4a R 4b , cyano, hydroxyl, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogenated C 1-4 Alkoxy, deuterated C 1-4 Alkyl, deuterated C 1-4 Alkoxy; R 4a and R 4b Each is selected from hydrogen, deuterium, C 1-6 Alkyl, C 3-6 Cycloalkyl, heterocyclyl, wherein the alkyl, cycloalkyl, heterocyclyl is optionally further substituted with one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R 5 Selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, mercapto, C 1-6 Alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are optionally further substituted with one or more substituents selected from deuterium, halogen, amino, oxo, thio, nitro, cyano, hydroxyl, mercapto, alkyl, deuterated alkyl, haloalkyl, alkoxy, deuterated alkoxy, haloalkoxy, hydroxyalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R 6 and R 7 are each independently selected from hydrogen, deuterium, halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; or R 6 and R 7 The bond to which it is attached, C, N, O or S together form a cycloalkyl group, a heterocyclic group, and the cyclic group is substituted by one or more groups selected from halogen, amino, nitro, cyano, hydroxyl, thiol, carboxyl, ester, alkyl, alkoxy, haloalkyl, hydroxyalkyl, aminoalkyl, haloalkoxy, alkenyl, alkynyl, cycloalkyl, heterocyclic group, aryl and heteroaryl.

2. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein A 1 and A 2 are C and CH, CH and C, C and N, N and C, CH and O, O and CH or C and C respectively.

3. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein X 1 C or N, X 2 and X 3 For C.

4. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein B 1 and B 2 Each is NR 5 and CR 5 , CR 5 and NR 5 , O and CR 5 , CR 5 and O, S and CR 5 , CR 5 and S, N and S, S and N, N and CR 5 , CR 5 and N or CR 5 and CR 5 .

5. The compound of general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein L1 is CH2.

6. The compound of claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein L2 is selected from a bond and O.

7. The compound of claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein R 1 and R 2 are each independently selected from hydrogen and C 1-6 Alkyl, or R 1 and R 2 Each does not exist; Or, R 1 With R 2 , A 1 , A 2 Together they form C 3-6 hydrocarbon ring, 3-6 membered heterocyclic ring containing 1-2 nitrogen, oxygen, sulfur atoms, C 6-10 aromatic ring or a 5-6 membered heteroaromatic ring containing 1-2 nitrogen, oxygen or sulfur atoms, wherein the hydrocarbon ring, heterocyclic ring, aromatic ring or heteroaromatic ring is optionally replaced by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 The cycloalkyl group is substituted with a substituent.

8. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein ring B is selected from a 4-6 membered saturated monocyclic heterocycle containing 1-2 nitrogen atoms and a 5-6 membered partially unsaturated monocyclic heterocycle containing 1-2 nitrogen atoms; m is 0, and R 3 Does not exist.

9. The compound represented by the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein ring B is selected from 10. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein ring C is selected from a 5-6 membered monocyclic aromatic heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms and an 8-10 membered bicyclic aromatic heterocycle containing 1-2 nitrogen, oxygen or sulfur atoms; n is 0, 1 or 2; each R 4 are each independently selected from halogen, C(O)NR 4a R 4b and C 1-6 Alkyl; R 4a and R 4b are each independently selected from hydrogen, C 1-6 Alkyl and deuterated C 1-6 alkyl.

11. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein Selected from 12. The compound of claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein R 5 Selected from hydrogen and C 1-6 alkyl.

13. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, which is a compound of the general formula (II) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt: in, Ring A is C 3-6 hydrocarbon ring, 3-6 membered heterocyclic ring containing 1-2 nitrogen, oxygen, sulfur atoms, C 6-10 aromatic ring or a 5-6 membered heteroaromatic ring containing 1-2 nitrogen, oxygen or sulfur atoms, wherein the hydrocarbon ring, heterocyclic ring, aromatic ring or heteroaromatic ring is optionally replaced by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 Substitution of cycloalkyl groups; A 1 , A 2 , X 1 , X 2 , X 3 , B 1 , B 2 , L1, L2, R 3 , R 4 , Ring B, Ring C, m and n are as defined in claim 1.

14. The compound represented by the general formula (I) according to claim 13 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein ring A is selected from wherein the ring A is optionally substituted by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 The cycloalkyl group is substituted with a substituent.

15. The compound represented by the general formula (I) according to claim 13 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, wherein ring A is selected from 16. The compound of the general formula (I) according to claim 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, which is a compound of the general formula (I-1) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt: in X 1 C or N; A 1 , A 2 , R 1 , R 2 , B 1 , B 2 , L2, R 4 , Ring B, Ring C and n are as defined in claim 1.

17. The compound of the general formula (I) according to claim 1, or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, which is a compound of the general formula (I-2) or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt: in Ring A is C 3-6 hydrocarbon ring, 3-6 membered heterocyclic ring containing 1-2 nitrogen, oxygen, sulfur atoms, C 6-10 aromatic ring or a 5-6 membered heteroaromatic ring containing 1-2 nitrogen, oxygen or sulfur atoms, wherein the hydrocarbon ring, heterocyclic ring, aromatic ring or heteroaromatic ring is optionally replaced by one or more selected from C 1-6 Alkyl, halogen, cyano or C 3-6 Substitution of cycloalkyl groups; X 1 C or N; A 1 , A 2 , B 1 , B 2 , L2, R 4 , Ring B, Ring C and n are as defined in claim 1.

18. The compound of the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, which is selected from the following compounds: or a tautomer, meso racemate, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt thereof.

19. A method for preparing the compound of general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, comprising the following steps: The compound represented by the general formula III or its salt undergoes an affinity substitution reaction with the compound represented by the general formula IV under alkaline conditions, optionally in the presence of a catalyst, to obtain a compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof; wherein Z is a halogen; A 1 , A 2 , X 1 , X 2 , X 3 , B 1 , B 2 , L1, L2, Ring B, Ring C, m and n are as defined in claim 1.

20. A method for preparing the compound of general formula (I) according to claim 13 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, comprising the following steps: The compound represented by the general formula V or its salt undergoes an affinity substitution reaction with the compound represented by the general formula IV under alkaline conditions, optionally in the presence of a catalyst, to obtain a compound represented by the general formula (II) or a pharmaceutically acceptable salt thereof; wherein Z is a halogen; A 1 , A 2 , X 1 , X 2 , X 3 , B 1 , B 2 , L1, L2, Ring A, Ring B, Ring C, m and n are as defined in claim 13.

21. A pharmaceutical composition comprising the compound represented by the general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier.

22. Use of the compound represented by general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 21 in the preparation of a PARP1 inhibitor.

23. Use of the compound represented by general formula (I) according to claim 1 or its tautomer, mesomer, racemate, enantiomer, diastereomer or pharmaceutically acceptable salt or the pharmaceutical composition according to claim 21 in the preparation of a medicament for preventing and / or treating diseases associated with PARP1 activity.

24. The use according to claim 23, wherein the disease associated with PARP1 activity is a solid tumor, such as ovarian cancer, breast cancer, prostate cancer, etc.

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