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CN106916101A - Double target spot inhibitor of NAMPT/HDAC and preparation method thereof - Google Patents

Double target spot inhibitor of NAMPT/HDAC and preparation method thereof Download PDF

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CN106916101A
CN106916101A CN201710082432.5A CN201710082432A CN106916101A CN 106916101 A CN106916101 A CN 106916101A CN 201710082432 A CN201710082432 A CN 201710082432A CN 106916101 A CN106916101 A CN 106916101A
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aminophenyls
methyl
benzamide
triazol
compound
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CN106916101B (en
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盛春泉
李宝力
董国强
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Second Military Medical University SMMU
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Poly (shanghai) Biological Technology Co Ltd
Second Military Medical University SMMU
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Abstract

本发明公开了一种NAMPT/HDAC双靶点抑制剂及其制备方法；该双靶点抑制剂为酰胺类衍生物及其药学上可接受的盐，其结构通式如式(I)所示：(I)。药理实验表明，本发明所述的衍生物或盐，对烟酰胺磷酸核糖转移酶和组蛋白去乙酰化酶均具有很强的抑制活性，而且具有较强的体外抗肿瘤活性和优秀的体内抑瘤效果。本发明还提供了上述衍生物及其药学上可接受的盐的制备方法，以及在制备烟酰胺磷酸核糖转移酶抑制剂、组蛋白去乙酰化酶抑制剂和抗肿瘤药物中的应用。

The invention discloses a NAMPT/HDAC dual-target inhibitor and a preparation method thereof; the dual-target inhibitor is an amide derivative and a pharmaceutically acceptable salt thereof, and its general structural formula is shown in formula (I) : (I). Pharmacological experiments show that the derivatives or salts of the present invention have strong inhibitory activity on nicotinamide phosphoribosyltransferase and histone deacetylase, and have strong antitumor activity in vitro and excellent inhibitory activity in vivo. tumor effect. The present invention also provides a preparation method of the above-mentioned derivatives and pharmaceutically acceptable salts thereof, as well as applications in the preparation of nicotinamide phosphoribosyl transferase inhibitors, histone deacetylase inhibitors and antitumor drugs.

Description

NAMPT/HDAC双靶点抑制剂及其制备方法NAMPT/HDAC dual-target inhibitor and preparation method thereof

技术领域technical field

本发明属于医药技术领域；具体涉及一类具有抗肿瘤活性的NAMPT/HDAC双靶点抑制剂及其药用制剂在治疗恶性肿瘤及与分化增殖相关疾病药物方面的应用。The invention belongs to the technical field of medicine; in particular, it relates to the application of a class of NAMPT/HDAC dual-target inhibitors with anti-tumor activity and their pharmaceutical preparations in the treatment of malignant tumors and drugs related to differentiation and proliferation.

背景技术Background technique

多靶点药物可以同时调节肿瘤疾病网络系统中的多个环节，不易产生耐药性，对各靶点作用的总效应大于单个效应之和，达到最佳的治疗效果。此外，单个分子拥有相对简单的吸收、分布、代谢和排泄过程，大大减少药物-药物相互作用(Peters，J.，etal.J.Med.Chem.2013，56，8955-8971)。FDA先后批准了多个多靶点酪氨酸激酶抑制剂上市，包括索拉非尼(sorafenib)、达沙替尼(dasatinib)和拉帕替尼(lapatinib)等，标志着多靶点药物已经成为肿瘤治疗和药物研发的新方向。Multi-target drugs can simultaneously regulate multiple links in the tumor disease network system, and are not prone to drug resistance. The total effect on each target is greater than the sum of individual effects, achieving the best therapeutic effect. In addition, single molecules possess relatively simple absorption, distribution, metabolism and excretion processes, greatly reducing drug-drug interactions (Peters, J., et al. J. Med. Chem. 2013, 56, 8955-8971). The FDA has successively approved the marketing of multiple multi-target tyrosine kinase inhibitors, including sorafenib, dasatinib and lapatinib, etc., marking that multi-target drugs have It has become a new direction for tumor treatment and drug development.

组蛋白的乙酰化/去乙酰化是染色体结构改变和基因表达的重要调节方式，在细胞凋亡、能量代谢、转录和翻译等生命过程中发挥重要作用。组蛋白的乙酰化和去乙酰化修饰过程主要由组蛋白乙酰化酶(histone acetylases，HATs)和组蛋白去乙酰化酶(histonedeacetylases，HDACs)共同催化完成。在前列腺癌、胃癌、乳腺癌、结肠癌和白血病等肿瘤细胞中，HDACs表达异常升高，针对HDAC开展小分子药物研发，已经成为目前抗肿瘤靶向治疗的热点。按结构类型HDAC抑制剂主要分为两类：异羟肟酸类和酰胺类。代表化合物分别为vorinostat(SAHA)和CI-994。SAHA经美国FDA批准进入临床用于治疗血液癌症；CI-994目前正在进行临床II期实验研究。此外，尚有多个HDAC抑制剂正在开展临床实验。然而，单一靶向HDAC的药物对实体瘤缺乏有效的治疗。HDAC与多个肿瘤靶点具有协同抗肿瘤的功效，例如微观蛋白(Tubulin)和热休克蛋白90(Hsp90)等，发展基于HDAC的多靶点药物成为提高抗肿瘤疗效、降低肿瘤耐药的有效手段。Acetylation/deacetylation of histone is an important regulation method of chromosome structure change and gene expression, and plays an important role in life processes such as apoptosis, energy metabolism, transcription and translation. Histone acetylation and deacetylation are mainly catalyzed by histone acetylases (HATs) and histone deacetylases (HDACs). In tumor cells such as prostate cancer, gastric cancer, breast cancer, colon cancer, and leukemia, the expression of HDACs is abnormally elevated. The development of small molecule drugs targeting HDACs has become a hot spot for anti-tumor targeted therapy. According to the structure type, HDAC inhibitors are mainly divided into two categories: hydroxamic acids and amides. The representative compounds are vorinostat (SAHA) and CI-994 respectively. SAHA has been approved by the US FDA for clinical use in the treatment of blood cancer; CI-994 is currently undergoing clinical phase II experimental research. In addition, there are several HDAC inhibitors in clinical trials. However, drugs targeting HDACs alone lack effective treatment for solid tumors. HDAC has synergistic anti-tumor effects with multiple tumor targets, such as Tubulin and heat shock protein 90 (Hsp90), etc. The development of HDAC-based multi-target drugs has become an effective strategy for improving anti-tumor efficacy and reducing tumor drug resistance. means.

近年来，针对代谢靶点开展药物研发已经成为抗肿瘤药物发现的重要手段和行之有效的策略。烟酰胺磷酸核糖转移酶(nicotinamidephosphoribosyltransferase，NAMPT)则是其中极具代表性的代谢类靶点。NAMPT催化烟酰胺(nicotinamide，NAM)生成烟酰胺单核苷酸(nicotinaminde mononucleotide，NMN)，调控哺乳动物细胞必需能量物质NAD的水平，是NAD生成途径的限速酶，在细胞生理活动中起着至关重要的作用。研究表明，NAMPT与肿瘤的发生和发展密切相关，已经成为抗肿瘤药物研究中一个非常重要的新靶点：1)肿瘤细胞具有很高的NAD消耗和代谢速率，肿瘤细胞比正常细胞对NAD的依赖性更强，更容易受到NAMPT抑制剂的影响；2)在肿瘤细胞中NAD作为必需的辅酶参与多种肿瘤必需物质的合成，而且，NAD能够显著降低环境中活性氧自由基(reactive oxygen species，ROS)水平，保护肿瘤细胞；3)NAMPT在血管新生、诱导血管内皮生长因子生成中起到重要的作用。目前有研究报道的NAMPT抑制剂有FK866和CHS-828，两者均已进入临床研究。临床数据显示，两者存在着剂量依赖的血小板减少症和胃肠道副作用，尚需要进一步改善。In recent years, drug development targeting metabolic targets has become an important means and effective strategy for anticancer drug discovery. Nicotinamide phosphoribosyltransferase (nicotinamidephosphoribosyltransferase, NAMPT) is one of the most representative metabolic targets. NAMPT catalyzes nicotinamide (nicotinamide, NAM) to generate nicotinamide mononucleotide (nicotinaminde mononucleotide, NMN), regulates the level of NAD, an essential energy substance in mammalian cells, is the rate-limiting enzyme of the NAD production pathway, and plays a role in cell physiological activities Crucial role. Studies have shown that NAMPT is closely related to the occurrence and development of tumors, and has become a very important new target in the research of anticancer drugs: 1) Tumor cells have a high NAD consumption and metabolic rate, and tumor cells have a higher NAD consumption than normal cells. 2) In tumor cells, NAD, as an essential coenzyme, participates in the synthesis of various tumor essential substances, and NAD can significantly reduce the reactive oxygen species in the environment (reactive oxygen species , ROS) levels, protecting tumor cells; 3) NAMPT plays an important role in angiogenesis and induction of vascular endothelial growth factor production. Currently, the NAMPT inhibitors reported in research include FK866 and CHS-828, both of which have entered clinical research. Clinical data show that both have dose-dependent thrombocytopenia and gastrointestinal side effects, which need further improvement.

发明内容Contents of the invention

本发明的目的在于提供一类酰胺类高效低毒的HDAC/NAMPT双靶点抑制剂及其药学上可接受的盐；本发明的另一目的是提供该类HDAC/NAMPT双靶点抑制剂及其药学上可接受的盐的制备方法；本发明的第三目的是提供该类HDAC/NAMPT双靶点抑制剂及其药学上可接受的盐在制备NAMPT抑制剂、HDAC抑制剂和抗肿瘤药物中的应用。The object of the present invention is to provide a class of amide-based high-efficiency and low-toxicity HDAC/NAMPT dual-target inhibitors and pharmaceutically acceptable salts thereof; another object of the present invention is to provide such HDAC/NAMPT dual-target inhibitors and The preparation method of its pharmaceutically acceptable salt; The third object of the present invention is to provide this kind of HDAC/NAMPT dual-target inhibitor and pharmaceutically acceptable salt thereof in the preparation of NAMPT inhibitor, HDAC inhibitor and antitumor drug in the application.

本发明的目的是通过以下技术方案来实现的：The purpose of the present invention is achieved through the following technical solutions:

本发明的第一方面，是提供一种酰胺类衍生物及其药学上可接受的盐，该化合物的结构通式如式(I)所示：The first aspect of the present invention is to provide an amide derivative and a pharmaceutically acceptable salt thereof, the general structural formula of which is shown in formula (I):

其中，A基团选自(1)～(4)中任意一种：Wherein, A group is selected from any one of (1)～(4):

(1)苯基或取代苯基(1) Phenyl or substituted phenyl

取代苯基苯环上的取代基可位于苯环的邻位、间位和对位，可以是单取代，也可以是多取代，取代基选自以下任意一种或几种：The substituents on the substituted phenyl benzene ring can be located in the ortho, meta and para positions of the benzene ring, and can be mono-substituted or multi-substituted, and the substituents can be selected from any one or more of the following:

a、卤素原子(包括F、Cl、Br、I)，a. Halogen atoms (including F, Cl, Br, I),

b、1-6个碳原子的直链烷基或3-6个碳原子的支链烷基，(优选的，所述1-6个碳原子的直链烷基包括甲基、乙基、丙基、正丁基、正戊基、正己基；所述3-6个碳原子的支链烷基包括异丙基、异丁基、叔丁基、异戊基、叔戊基)，b. Straight-chain alkyl groups of 1-6 carbon atoms or branched-chain alkyl groups of 3-6 carbon atoms, (preferably, the straight-chain alkyl groups of 1-6 carbon atoms include methyl, ethyl, Propyl, n-butyl, n-pentyl, n-hexyl; the branched chain alkyl group with 3-6 carbon atoms includes isopropyl, isobutyl, tert-butyl, isopentyl, tert-amyl),

c、羟基、氰基、氰基甲基、氨基、硝基、三氟甲基、三氟甲氧基、甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、乙酰基；c, hydroxyl, cyano, cyanomethyl, amino, nitro, trifluoromethyl, trifluoromethoxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, acetyl base;

(2)吡啶基或取代吡啶基(2) pyridyl or substituted pyridyl

取代吡啶上取代基可位于吡啶环任何可取代位置，可以是单取代，也可以是多取代，取代基选自以下任意一种或几种：The substituent on the substituted pyridine can be located at any substitutable position of the pyridine ring, and can be single-substituted or multi-substituted, and the substituent can be selected from any one or more of the following:

a、卤素原子(包括F、Cl、Br、I)，a. Halogen atoms (including F, Cl, Br, I),

b、1-6个碳原子的直链烷基或3-6个碳原子的支链烷基，(优选的，所述1-6个碳原子的直链烷基包括甲基、乙基、丙基、正丁基、正戊基、正己基；所述3-6个碳原子的支链烷基包括异丙基、异丁基、叔丁基、异戊基、叔戊基)，b. Straight-chain alkyl groups of 1-6 carbon atoms or branched-chain alkyl groups of 3-6 carbon atoms, (preferably, the straight-chain alkyl groups of 1-6 carbon atoms include methyl, ethyl, Propyl, n-butyl, n-pentyl, n-hexyl; the branched chain alkyl group with 3-6 carbon atoms includes isopropyl, isobutyl, tert-butyl, isopentyl, tert-amyl),

c、羟基、氰基、氰基甲基、氨基、硝基、三氟甲基、三氟甲氧基、甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、乙酰基；c, hydroxyl, cyano, cyanomethyl, amino, nitro, trifluoromethyl, trifluoromethoxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, acetyl base;

(3)五元杂环或取代五元杂环(五元杂环指呋喃、噻吩、吡咯基因)(3) Five-membered heterocycle or substituted five-membered heterocycle (five-membered heterocycle refers to furan, thiophene, and pyrrole genes)

五元杂环上的取代基可位于任何可取代位置，可以是单取代，也可以是多取代，取代基选自以下任意一种或几种：The substituents on the five-membered heterocyclic ring can be located in any substitutable position, and can be mono-substituted or multi-substituted, and the substituents can be selected from any one or more of the following:

a、卤素原子(包括F、Cl、Br、I)，a. Halogen atoms (including F, Cl, Br, I),

b、1-6个碳原子的直链烷基或3-6个碳原子的支链烷基，(优选的，所述1-6个碳原子的直链烷基包括甲基、乙基、丙基、正丁基、正戊基、正己基；所述3-6个碳原子的支链烷基包括异丙基、异丁基、叔丁基、异戊基、叔戊基)，b. Straight-chain alkyl groups of 1-6 carbon atoms or branched-chain alkyl groups of 3-6 carbon atoms, (preferably, the straight-chain alkyl groups of 1-6 carbon atoms include methyl, ethyl, Propyl, n-butyl, n-pentyl, n-hexyl; the branched chain alkyl group with 3-6 carbon atoms includes isopropyl, isobutyl, tert-butyl, isopentyl, tert-amyl),

c、羟基、氰基、氰基甲基、氨基、硝基、三氟甲基、三氟甲氧基、甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、乙酰基；c, hydroxyl, cyano, cyanomethyl, amino, nitro, trifluoromethyl, trifluoromethoxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, acetyl base;

(4)五元或六元环组成的骈基和二环杂环基；(4) Heterocyclyl and bicyclic heterocyclyl composed of five-membered or six-membered rings;

E基团选自以下基团中的一种：The E group is selected from one of the following groups:

X基团选自以下基团中的一种：The X group is selected from one of the following groups:

其中，R1指的是单取代或多取代，选自以下任意一种或几种：Among them, R1 refers to single substitution or multiple substitution, selected from any one or more of the following:

a.H，a.H,

b.卤素原子(F、Cl、Br、I)，b. Halogen atoms (F, Cl, Br, I),

g.1-6个碳原子的直链烷基或3-6个碳原子的支链烷基，(优选的，所述1-6个碳原子的直链烷基包括甲基、乙基、丙基、正丁基、正戊基、正己基；所述3-6个碳原子的支链烷基包括异丙基、异丁基、叔丁基、异戊基、叔戊基)，g. 1-6 straight-chain alkyl groups of carbon atoms or branched-chain alkyl groups of 3-6 carbon atoms, (preferably, said 1-6 carbon-atom straight-chain alkyl groups include methyl, ethyl, Propyl, n-butyl, n-pentyl, n-hexyl; the branched chain alkyl group with 3-6 carbon atoms includes isopropyl, isobutyl, tert-butyl, isopentyl, tert-amyl),

h.羟基、氰基、氰基甲基、氨基、硝基、三氟甲基、三氟甲氧基、甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、乙酰基；h. Hydroxy, cyano, cyanomethyl, amino, nitro, trifluoromethyl, trifluoromethoxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, acetyl base;

Y为碳原子或者氮原子；Y is a carbon atom or a nitrogen atom;

Z基团选自a～d中任意一种：The Z group is selected from any of a to d:

a.Ha.H

b.卤素原子(F、Cl、Br、I)b. Halogen atoms (F, Cl, Br, I)

c.苯环或六元杂环c. Benzene ring or six-membered heterocycle

d.五元杂环(包括吡咯、呋喃、噻吩基团)d. Five-membered heterocycles (including pyrrole, furan, thiophene groups)

优选的，所述五元或六元环组成的骈基和二环杂环基，包含：Preferably, the parallel group and bicyclic heterocyclic group composed of five-membered or six-membered rings include:

优选的，所述的药学上可接受的盐，较优的是：盐酸盐、氢溴酸盐、硫酸盐、乙酸盐、乳酸盐、酒石酸盐、鞣酸盐、枸橼酸盐、三氟醋酸盐、苹果酸盐、马来酸盐、琥珀酸盐、对甲苯磺酸盐或甲磺酸盐；Preferably, the pharmaceutically acceptable salts are preferably: hydrochloride, hydrobromide, sulfate, acetate, lactate, tartrate, tannate, citrate, Trifluoroacetate, malate, maleate, succinate, p-toluenesulfonate, or mesylate;

优选的，所述的药学上可接受的盐，不含结晶水，或含一个或一个以上分子的结晶水。Preferably, the pharmaceutically acceptable salt does not contain water of crystallization, or contains water of crystallization of one or more molecules.

更优选的，所述的药学上可接受的盐含0.5-3.0分子的结晶水。More preferably, said pharmaceutically acceptable salt contains 0.5-3.0 molecules of crystal water.

具有分化和抗增殖活性的HDAC抑制剂与HDAC/NAMPT双靶点抑制剂及其药用制剂，本发明部分优选化合物包括，但不限于以下：HDAC inhibitors with differentiation and anti-proliferation activities, HDAC/NAMPT dual-target inhibitors and pharmaceutical preparations thereof, some preferred compounds of the present invention include, but are not limited to the following:

N-(2-氨基苯基)-4-(3-(6-氟吡啶-3-基)甲基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(6-fluoropyridin-3-yl)methyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(吡啶-3-基甲基)脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(pyridin-3-ylmethyl)ureido)benzamide

N-(2-氨基苯基)-4-(3-(吡啶-3-基)丙烯酰胺基)苯甲酰胺N-(2-aminophenyl)-4-(3-(pyridin-3-yl)acrylamido)benzamide

N-(2-氨基苯基)-4-(3-(6-氯吡啶-3-基)甲基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(6-chloropyridin-3-yl)methyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(6-氯吡啶-3-基)甲基)脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(6-chloropyridin-3-yl)methyl)ureido)benzamide

N-(2-氨基苯基)-4-(3-(6-氟吡啶-3-基)甲基)脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(6-fluoropyridin-3-yl)methyl)ureido)benzamide

N-(2-氨基苯基)-4-(3-(6-氨基吡啶-3-基)甲基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(6-aminopyridin-3-yl)methyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(6-氨基吡啶-3-基)甲基)脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(6-aminopyridin-3-yl)methyl)ureido)benzamide

叔丁基(5-((3-(4-((2-氨基苯基)氨基甲酰)苯基)脲基)甲基)吡啶-2-基)氨基甲酸酯tert-Butyl(5-((3-(4-((2-aminophenyl)carbamoyl)phenyl)ureido)methyl)pyridin-2-yl)carbamate

N-(2-氨基苯基)-4-(3-(5-氟吡啶-3-基)甲基)脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(5-fluoropyridin-3-yl)methyl)ureido)benzamide

N-(2-氨基苯基)-4-(3-(6-甲基吡啶-3-基)甲基)脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(6-methylpyridin-3-yl)methyl)ureido)benzamide

N-(2-氨基苯基)-4-(3-(4-氟苄基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(4-fluorobenzyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(3-氟苄基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(3-fluorobenzyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(2-氟苄基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(2-fluorobenzyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(3-氯苄基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(3-chlorobenzyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(3-溴苄基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(3-bromobenzyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(苯并[d][1，3]二氧杂环戊烯-5-基甲基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(benzo[d][1,3]dioxol-5-ylmethyl)thioureido)benzamide

N-(2-氨基苯基)-4-(3-(3-氨基苄基)硫脲基)苯甲酰胺N-(2-aminophenyl)-4-(3-(3-aminobenzyl)thioureido)benzamide

4-(3-((1H-苯并[d]咪唑-2-基)甲基)硫脲基)-N-(2-氨基苯基)苯甲酰胺4-(3-((1H-Benzo[d]imidazol-2-yl)methyl)thioureido)-N-(2-aminophenyl)benzamide

N-(2-氨基苯基)-4-(2，3-二氯-3-(吡啶-3-基)丙烯酰胺基)苯甲酰胺N-(2-aminophenyl)-4-(2,3-dichloro-3-(pyridin-3-yl)acrylamido)benzamide

N-(4-((2-氨基苯基)氨基甲酰基)苄基)噻吩并[3，2-c]吡啶-2-甲酰胺N-(4-((2-aminophenyl)carbamoyl)benzyl)thieno[3,2-c]pyridine-2-carboxamide

N-(4-((2-氨基苯基)氨基甲酰)苄基)-1H-吡咯[2，3-c]吡啶-2-甲酰胺N-(4-((2-aminophenyl)carbamoyl)benzyl)-1H-pyrrole[2,3-c]pyridine-2-carboxamide

N-(4-((2-氨基苯基)氨基甲酰)苄基)-1H-吡咯并[3，2-c]吡啶-2-甲酰胺N-(4-((2-aminophenyl)carbamoyl)benzyl)-1H-pyrrolo[3,2-c]pyridine-2-carboxamide

N-(4-((2-氨基苯基)氨基甲酰)苄基)-1H-吡咯并[2，3-b]吡啶-2-甲酰胺N-(4-((2-aminophenyl)carbamoyl)benzyl)-1H-pyrrolo[2,3-b]pyridine-2-carboxamide

N-(4-((2-氨基苯基)氨基甲酰基)苄基)咪唑并[1，5-a]吡啶-6-甲酰胺N-(4-((2-aminophenyl)carbamoyl)benzyl)imidazo[1,5-a]pyridine-6-carboxamide

N-(4-((2-氨基苯基)氨基甲酰基)苄基)-1H-吡唑并[4，3-b]吡啶-6-甲酰胺N-(4-((2-aminophenyl)carbamoyl)benzyl)-1H-pyrazolo[4,3-b]pyridine-6-carboxamide

N-(4-((2-氨基苯基)氨基甲酰基)苄基)咪唑[1，2-a]吡啶-7-甲酰胺N-(4-((2-aminophenyl)carbamoyl)benzyl)imidazol[1,2-a]pyridine-7-carboxamide

N-(2-氨基苯基)-4-((4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)methyl)benzamide

N-(2-氨基苯基)-4-(2-(4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-(2-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)methyl)benzamide

N-(2-氨基苯基)-4-((4-(吡啶-4-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(pyridin-4-yl)-1H-1,2,3-triazol-1-yl)methyl)benzamide

N-(2-氨基苯基)-4-((4-(6-氯吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(6-chloropyridin-3-yl)-1H-1,2,3-triazol-1-yl)methyl)benzamide

N-(2-氨基苯基)-4-((4-(3-氨基苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)methyl)benzamide

N-(2-氨基苯基)-4-((4-(间甲苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(m-tolyl)-1H-1,2,3-triazol-1-yl)methyl)benzamide

4-((4-(4-乙酰基苯基)-1H-1，2，3-三唑-1-基)甲基)-N-(2-氨基苯基)苯甲酰胺4-((4-(4-acetylphenyl)-1H-1,2,3-triazol-1-yl)methyl)-N-(2-aminophenyl)benzamide

N-(2-氨基苯基)-4-((4-(噻吩-2-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(thiophen-2-yl)-1H-1,2,3-triazol-1-yl)methyl)benzamide

N-(4-氨-[1，1′-联苯]-3-基)-4-((5-(吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(4-amino-[1,1'-biphenyl]-3-yl)-4-((5-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl ) methyl) benzamide

N-(4-氨基-[1，1′-联苯]-3-基)-4-((5-(3-氨基苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(4-amino-[1,1'-biphenyl]-3-yl)-4-((5-(3-aminophenyl)-1H-1,2,3-triazol-1-yl ) methyl) benzamide

N-(4-氨基-[1，1′-联苯]-3-基)-4-((5-(3-羟基苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(4-amino-[1,1'-biphenyl]-3-yl)-4-((5-(3-hydroxyphenyl)-1H-1,2,3-triazol-1-yl ) methyl) benzamide

4-((5-(4-乙酰基苯基)-1H-1，2，3-三唑-1-基)甲基)-N-(4-氨基-[1，1′-联苯]-3-基)苯甲酰胺4-((5-(4-acetylphenyl)-1H-1,2,3-triazol-1-yl)methyl)-N-(4-amino-[1,1′-biphenyl] -3-yl)benzamide

N-(2-氨基苯基)-4-((4-(4-(3-(吡啶-3-基甲基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(4-(3-(pyridin-3-ylmethyl)-1H-1,2,3-triazol-1-yl)methyl) benzamide

N-(2-氨基苯基)-4-((4-(4-((吡啶-3-基甲基)氨甲酰基)苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺N-(2-aminophenyl)-4-((4-(4-((pyridin-3-ylmethyl)carbamoyl)phenyl)-1H-1,2,3-triazole-1- base) methyl) benzamide

N-(2-氨基苯基)-8-(4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)辛酰胺N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)octylamide

N-(2-氨基苯基)-8-(4-(3-氨基苯基)-1H-1，2，3-三唑-1-基)辛酰胺N-(2-aminophenyl)-8-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl)octylamide

N-(4-氨基-[1，1′-联苯]-3-基)-8-(4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)辛酰胺N-(4-amino-[1,1′-biphenyl]-3-yl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl) Caprylamide

N-(4-氨基-[1，1′-联苯]-3-基)-8-(4-(3-氨基苯基)-1H-1，2，3-三唑-1-基)辛酰胺N-(4-amino-[1,1′-biphenyl]-3-yl)-8-(4-(3-aminophenyl)-1H-1,2,3-triazol-1-yl) Caprylamide

N-(4-氨基-[1，1′-联苯]-3-基)-8-(4-(3-羟基苯基)-1H-1，2，3-三唑-1-基)辛酰胺N-(4-amino-[1,1′-biphenyl]-3-yl)-8-(4-(3-hydroxyphenyl)-1H-1,2,3-triazol-1-yl) Caprylamide

上述本发明优选化合物的结构式和核磁质谱数据如表1所示：The structural formula and nuclear magnetic mass spectrum data of above-mentioned preferred compound of the present invention are as shown in table 1:

表1、本发明优选化合物的结构式和核磁质谱数据Table 1. Structural formula and NMR mass spectrometry data of preferred compounds of the present invention

本发明的第二方面，是提供上述酰胺类衍生物及其药学上可接受的盐的制备方法；The second aspect of the present invention is to provide the preparation method of the above-mentioned amide derivatives and pharmaceutically acceptable salts thereof;

反应流程通法一(包括化合物A1-20的合成：)Reaction scheme general method one (including the synthesis of compound A1-20:)

当结构通式为且X2为O或S，R为-NH-CH2-A、-CH＝CH-A或A为苯基、氨基取代苯基、卤素取代苯基、吡啶基、1-6个碳原子的直链烷基取代吡啶基、3-6个碳原子的支链烷基取代吡啶基、卤素取代吡啶基、氨基取代吡啶基、时，所述方法如下：When the general structure is And X 2 is O or S, R is -NH-CH 2 -A, -CH=CH-A or A is phenyl, amino-substituted phenyl, halogen-substituted phenyl, pyridyl, linear alkyl-substituted pyridyl with 1-6 carbon atoms, branched-chain alkyl-substituted pyridyl with 3-6 carbon atoms, halogen-substituted Pyridyl, amino-substituted pyridyl, , the method is as follows:

(其中，R2为氢、卤素、氨基或叔丁氧羰基氨基)(wherein, R2 is hydrogen, halogen, amino or tert-butoxycarbonylamino)

试剂和条件：(a)兰尼镍，氢气，氨水甲醇溶液，室温，12小时，产率21-89％；(b)CDIor TCDI，二氯甲烷，室温，2小时，产率85-95％；(c)四氢呋喃，室温，12小时，产率62-81％；(d)二氯甲烷，室温或70度，2小时，产率60％；(e)四氢呋喃，室温，12小时，产率70％；(f)邻苯二胺，HBTU，三乙胺，DMF，室温，3小时，收率43-75％.Reagents and conditions: (a) Raney nickel, hydrogen, ammonia methanol solution, room temperature, 12 hours, yield 21-89%; (b) CDIor TCDI, dichloromethane, room temperature, 2 hours, yield 85-95% (c) tetrahydrofuran, room temperature, 12 hours, yield 62-81%; (d) dichloromethane, room temperature or 70 degrees, 2 hours, yield 60%; (e) tetrahydrofuran, room temperature, 12 hours, yield 70%; (f) o-phenylenediamine, HBTU, triethylamine, DMF, room temperature, 3 hours, yield 43-75%.

氰基化合物1经兰尼镍催化，在氨水甲醇溶液中氢气还原得到氨基化合物2a。对氨基苯甲酸3与CDI或TCDI室温下反应得到中间体4。化合物2a或2c和4在四氢呋喃中室温反应12小时得到中间体5。化合物2b与3在二氯甲烷中缩合得到中间体5。中间体5在DMF中与邻苯二胺室温反应4小时，经柱层析得到目标化合物A1-20。The cyano compound 1 was catalyzed by Raney nickel, and the amino compound 2a was obtained by hydrogen reduction in ammonia-water-methanol solution. Reaction of p-aminobenzoic acid 3 with CDI or TCDI at room temperature gives intermediate 4. Compound 2a or 2c and 4 were reacted in tetrahydrofuran at room temperature for 12 hours to obtain intermediate 5. Compounds 2b and 3 were condensed in dichloromethane to give intermediate 5. Intermediate 5 was reacted with o-phenylenediamine in DMF for 4 hours at room temperature, and the target compound A1-20 was obtained by column chromatography.

反应流程通法二(含化合物B1-7的合成)：Reaction scheme general method 2 (comprising the synthesis of compound B1-7):

当结构通式为且A为 时，所述方法如下：When the general structure is and A is , the method is as follows:

试剂和条件：(a)R-COOH，EDC，DMAP，DMF，室温，3小时，产率31-69％；(b)氢氧化锂，四氢呋喃/甲醇/水，室温，24小时，产率82-97％；(c)邻苯二胺，HBTU，三乙胺，DMF，室温，4小时，收率37-75％.Reagents and conditions: (a) R-COOH, EDC, DMAP, DMF, room temperature, 3 hours, yield 31-69%; (b) lithium hydroxide, tetrahydrofuran/methanol/water, room temperature, 24 hours, yield 82% -97%; (c) o-phenylenediamine, HBTU, triethylamine, DMF, room temperature, 4 hours, yield 37-75%.

4-胺甲基苯甲酸甲酯7与各种羧酸经EDC/DMAP催化缩合，得到中间体8。在四氢呋喃/甲醇/水混合溶剂体系中，中间体8经氢氧化锂水解得到相应的羧酸化合物9。化合物9在DMF中与邻苯二胺在室温条件下反应3小时，柱层析得到目标化合物B1-7。EDC/DMAP catalyzed condensation of methyl 4-aminomethylbenzoate 7 with various carboxylic acids gave intermediate 8. In the tetrahydrofuran/methanol/water mixed solvent system, the intermediate 8 was hydrolyzed with lithium hydroxide to obtain the corresponding carboxylic acid compound 9. Compound 9 was reacted with o-phenylenediamine in DMF for 3 hours at room temperature, and the target compound B1-7 was obtained by column chromatography.

反应流程通法三(含化合物C1-14的合成)：Reaction scheme general method three (synthesis containing compound C1-14):

当结构通式为且n＝1或2，A为吡啶基、卤素取代吡啶基、氨基取代吡啶基、1-6个碳原子直链烷基取代苯基、乙酰基取代苯基、噻吩基、Z为氢原子或苯基时，所述方法如下：When the general structure is And n=1 or 2, A is pyridyl, halogen substituted pyridyl, amino substituted pyridyl, 1-6 carbon atoms linear alkyl substituted phenyl, acetyl substituted phenyl, thienyl, When Z is a hydrogen atom or a phenyl group, the method is as follows:

试剂和条件：(a)三甲基硅基乙炔，三苯基磷钯，碘化铜，三乙胺，室温，1小时；(b)碳酸钾，甲醇，室温，0.5小时，两步产率32-81％；(c)叠氮化钠，室温，8小时，产率64-85％；(d)中间体13或RCN，硫酸铜，抗坏血酸钠，氮气保护，室温，过夜，产率64-87％；(e)氢氧化锂，四氢呋喃/甲醇/水，室温，24小时，产率71-92％；(f)邻苯二胺或[1，1′-联苯]-3，4-二胺，HBTU，三乙胺，室温，3小时，产率27-72％.Reagents and conditions: (a) Trimethylsilylacetylene, triphenylphosphopalladium, copper iodide, triethylamine, room temperature, 1 hour; (b) potassium carbonate, methanol, room temperature, 0.5 hour, two-step yield 32-81%; (c) sodium azide, room temperature, 8 hours, yield 64-85%; (d) intermediate 13 or RCN, copper sulfate, sodium ascorbate, nitrogen protection, room temperature, overnight, yield 64% -87%; (e) lithium hydroxide, tetrahydrofuran/methanol/water, room temperature, 24 hours, yield 71-92%; (f) o-phenylenediamine or [1,1'-biphenyl]-3,4 -Diamine, HBTU, triethylamine, room temperature, 3 hours, yield 27-72%.

化合物11与三甲基硅基乙炔反应得化合物12，在碱性条件下室温脱三甲基硅基制得化合物13。溴化物14经叠氮化钠取代得到中间体15。中间体15与13或其他炔基化合物发生click反应得到产物16。中间体16在碱性条件下水解，与邻苯二胺或4-苯基邻苯二胺缩合最终获得目标化和物C1-14。Compound 11 was reacted with trimethylsilylacetylene to obtain compound 12, and compound 13 was obtained by removing trimethylsilyl at room temperature under basic conditions. Substitution of bromide 14 with sodium azide affords intermediate 15. The click reaction of intermediate 15 with 13 or other alkynyl compounds affords product 16. The intermediate 16 is hydrolyzed under alkaline conditions, and condensed with o-phenylenediamine or 4-phenyl-o-phenylenediamine to finally obtain the target compound C1-14.

反应流程通法四(含化合物D1-5的合成)：Reaction scheme general method four (comprising the synthesis of compound D1-5):

当通式结构为且R’为氨基、羟基或氢原子，X3为N或CH，Z为氢原子或苯基时，所述方法如下：When the general structure is And R' is an amino group, a hydroxyl group or a hydrogen atom, X3 is N or CH, and when Z is a hydrogen atom or a phenyl group, the method is as follows:

试剂和条件：(a)叠氮化钠，DMF，85度，6小时；(b)邻苯二胺或[1，1′-联苯]-3，4-二胺，HBTU，三乙胺，室温，3小时；(c)硫酸铜，抗坏血酸钠，叔丁醇∶水＝5∶1，室温，氮气，24小时。Reagents and conditions: (a) sodium azide, DMF, 85 degrees, 6 hours; (b) o-phenylenediamine or [1,1'-biphenyl]-3,4-diamine, HBTU, triethylamine , room temperature, 3 hours; (c) copper sulfate, sodium ascorbate, tert-butanol: water = 5:1, room temperature, nitrogen, 24 hours.

化合物18与叠氮化钠反应得到中间体19，19与邻苯二胺或4-苯基邻苯二胺在HBTU和三乙胺催化下发生缩合反应得到中间体20。中间体20用含有不同取代基的芳基乙炔发生Click反应得到化合物D1-5。Compound 18 was reacted with sodium azide to obtain intermediate 19, and 19 was condensed with o-phenylenediamine or 4-phenyl-o-phenylenediamine under the catalysis of HBTU and triethylamine to obtain intermediate 20. Intermediate 20 is subjected to Click reaction with aryl acetylenes containing different substituents to obtain compound D1-5.

本发明的第三方面，是提供上述的酰胺类衍生物及其药学上可接受的盐在制备烟酰胺磷酸核糖转移酶抑制剂、组蛋白去乙酰化酶抑制剂或HDAC/NAMPT双靶点抑制剂中的用途。The third aspect of the present invention is to provide the above-mentioned amide derivatives and pharmaceutically acceptable salts thereof in the preparation of nicotinamide phosphoribosyltransferase inhibitors, histone deacetylase inhibitors or HDAC/NAMPT dual-target inhibitors use in pharmaceuticals.

本发明的化合物经NAMPT和HDAC1活性抑制实验，证明大部分化合物具有较好的NAMPT和HDAC1抑制活性，A2、A3、C1、C2、C9、C13、C14、D1、D2、D3对NAMPT和HDAC1具有平衡抑制活性。The compounds of the present invention are tested for NAMPT and HDAC1 activity inhibition, and it is proved that most of the compounds have better NAMPT and HDAC1 inhibitory activity, and A2, A3, C1, C2, C9, C13, C14, D1, D2, D3 have NAMPT and HDAC1 inhibitory activity. Balance inhibitory activity.

本发明还提供了上述的酰胺类衍生物及其药学上可接受的盐在制备抗肿瘤药物中的用途。The present invention also provides the use of the above-mentioned amide derivatives and pharmaceutically acceptable salts thereof in the preparation of antitumor drugs.

优选的，所述的肿瘤为肝癌、肺癌、肠癌、卵巢癌、前列腺癌、胃癌、淋巴癌等。Preferably, the tumor is liver cancer, lung cancer, colon cancer, ovarian cancer, prostate cancer, gastric cancer, lymphatic cancer and the like.

本发明的化合物经抗肿瘤活性实验，证明大部分化合物具有较好体外抗肿瘤活性，优选化合物具有优秀的体内肿瘤生长抑制活性。The anti-tumor activity test of the compounds of the present invention proves that most of the compounds have good anti-tumor activity in vitro, and the preferred compounds have excellent tumor growth inhibitory activity in vivo.

与现有技术相比，本发明具有如下有益效果：Compared with the prior art, the present invention has the following beneficial effects:

本发明通过药效团融合策略合理构建酰胺类HDAC/NAMPT双靶点抑制剂，获得对NAMPT和HDAC1具有平衡抑制活性的高效小分子，实现体内药效的显著提高；为深入研究和开发新结构类型抗肿瘤药物开辟了新的途径，提供了新的策略。The present invention rationally constructs amide-type HDAC/NAMPT dual-target inhibitors through a pharmacophore fusion strategy, obtains a high-efficiency small molecule with balanced inhibitory activity on NAMPT and HDAC1, and realizes a significant improvement in drug efficacy in vivo; for in-depth research and development of new structures Types of antineoplastic drugs open up new avenues and provide new strategies.

附图说明Description of drawings

通过阅读参照以下附图对非限制性实施例所作的详细描述，本发明的其它特征、目的和优点将会变得更明显：Other characteristics, objects and advantages of the present invention will become more apparent by reading the detailed description of non-limiting embodiments made with reference to the following drawings:

图1为人体肠癌HCT116裸鼠瘤体积变化示意图；其中，(A)肿瘤体积折线图；(B)给药结束后肿瘤解剖图。Fig. 1 is a schematic diagram of the tumor volume change of human intestinal cancer HCT116 in nude mice; wherein, (A) a line graph of tumor volume; (B) an anatomical diagram of the tumor after administration.

具体实施方式detailed description

下面结合实施例对本发明进行详细说明。以下实施例将有助于本领域的技术人员进一步理解本发明，但不以任何形式限制本发明。应当指出的是，对本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干调整和改进。这些都属于本发明的保护范围。本发明所述的百分比出特别说明外，均为重量百分比。The present invention will be described in detail below in conjunction with examples. The following examples will help those skilled in the art to further understand the present invention, but do not limit the present invention in any form. It should be noted that those skilled in the art can make some adjustments and improvements without departing from the concept of the present invention. These all belong to the protection scope of the present invention. Unless otherwise specified, the percentages described in the present invention are all percentages by weight.

实施例1、N-(2-氨基苯基)-4-(3-(6-氟吡啶-3-基)甲基)硫脲基)苯甲酰胺制备Embodiment 1, N-(2-aminophenyl)-4-(3-(6-fluoropyridin-3-yl)methyl)thioureido)benzamide preparation (A1)(A1)

1.1制备中间体2：6-氟-3-氨甲基吡啶1.1 Preparation of intermediate 2: 6-fluoro-3-aminomethylpyridine

将6-氟-3-氰基吡啶(300mg，2.46mmol)溶于的2mol/L氨/甲醇溶液中(30mL)，加入兰尼镍(0.5g)，H2氛围下室温反应12小时。反应完后反应液硅藻土过滤，甲醇冲洗(5mL×2)，滤液蒸干，残留物经硅胶柱层析(二氯甲烷∶甲醇＝100∶4)分离纯化，得到黄色油状固体262mg，收率为85％。1H-NMR(DMSO-d6，300MHz)δ：8.11-6.84(m，3H)，3.86(s，2H)，1.47(s，2H).6-Fluoro-3-cyanopyridine (300mg, 2.46mmol) was dissolved in 2mol/L ammonia/methanol solution (30mL), added Raney nickel (0.5g), and reacted at room temperature under H 2 atmosphere for 12 hours. After the reaction, the reaction solution was filtered with celite, rinsed with methanol (5mL×2), the filtrate was evaporated to dryness, and the residue was separated and purified by silica gel column chromatography (dichloromethane:methanol=100:4) to obtain 262 mg of a yellow oily solid, which was collected as The rate is 85%. 1 H-NMR (DMSO-d 6 , 300MHz) δ: 8.11-6.84 (m, 3H), 3.86 (s, 2H), 1.47 (s, 2H).

1.2制备中间体4：4-异硫氰基苯甲酸1.2 Preparation of intermediate 4: 4-isothiocyanatobenzoic acid

三乙胺(1.4mL，194mmol)溶于二氯甲烷(12mL)，冰浴下加入硫代羰基二咪唑(2.11g，11.85mmol)和4-氨基苯甲酸(1.25g，11.57mmol)，冰浴下搅拌1小时。浓盐酸(3mL，33.60mmol)滴加至正己烷(12mL)中制得混合溶液，将该混合溶液缓慢滴加至反应液中，室温搅拌3小时。反应完后将反应液置于冰浴中，过滤，滤饼水洗(5mL×2)，得到灰白色固体1.93g，收率为93％。1H-NMR(DMSO-d6，300MHz)δ：13.20(br s，1H)，7.52(d，J＝8.7Hz，2H)，7.97(d，J＝8.7Hz，2H).Triethylamine (1.4mL, 194mmol) was dissolved in dichloromethane (12mL), and thiocarbonyldiimidazole (2.11g, 11.85mmol) and 4-aminobenzoic acid (1.25g, 11.57mmol) were added under ice-cooling. Stir for 1 hour. Concentrated hydrochloric acid (3 mL, 33.60 mmol) was added dropwise to n-hexane (12 mL) to prepare a mixed solution, which was slowly added dropwise to the reaction solution, and stirred at room temperature for 3 hours. After the reaction, the reaction solution was placed in an ice bath, filtered, and the filter cake was washed with water (5 mL×2) to obtain 1.93 g of an off-white solid with a yield of 93%. 1 H-NMR (DMSO-d 6 , 300MHz) δ: 13.20 (br s, 1H), 7.52 (d, J=8.7Hz, 2H), 7.97 (d, J=8.7Hz, 2H).

1.3制备中间体5：4-(3-((6-氟吡啶-3-基)甲基)硫脲基)苯甲酸1.3 Preparation of intermediate 5: 4-(3-((6-fluoropyridin-3-yl)methyl)thioureido)benzoic acid

6-氟-3-氨甲基吡啶(133mg，1.06mmol)和4-异硫氰基苯甲酸(189mg，1.06mmol)溶于干燥THF(10mL)，室温条件下反应12小时，反应完后蒸干溶剂得到黄色固体0.26g，收率81％。1H-NMR(DMSO-d6，300MHz)δ：10.24(s，1H)，8.75(s，1H)，8.36-8.34(m，2H)，7.87(d，J＝8.5Hz，2H)，7.63-7.58(m，3H)，4.80(d，J＝5.4Hz，2H).ESI-MS(m/s)：287.12[M+1].6-fluoro-3-aminomethylpyridine (133mg, 1.06mmol) and 4-isothiocyanatobenzoic acid (189mg, 1.06mmol) were dissolved in dry THF (10mL), and reacted at room temperature for 12 hours. After drying the solvent, 0.26 g of a yellow solid was obtained, with a yield of 81%. 1 H-NMR (DMSO-d 6 , 300MHz) δ: 10.24 (s, 1H), 8.75 (s, 1H), 8.36-8.34 (m, 2H), 7.87 (d, J=8.5Hz, 2H), 7.63 -7.58(m, 3H), 4.80(d, J=5.4Hz, 2H).ESI-MS(m/s): 287.12[M+1].

1.4制备目标产物A11.4 Preparation of target product A1

4-(3-((6-氟吡啶-3-基)甲基)硫脲基)苯甲酸(90mg，0.30mmol)溶于干燥DMF(5mL)，加入邻苯二胺(32mg，0.30mmol)、HBTU(114mg，0.30mmol)，三乙胺(41μL，0.30mmol)，室温反应4小时。反应后将反应液倒入水中(40mL)，过滤得到粗品。粗品经柱层析(二氯甲烷∶甲醇＝100∶3)得到白色粉末59mg，收率51％。1H-NMR(DMSO-d6，300MHz)δ：9.95(s，1H)，9.56(s，1H)，8.46(s，1H)，8.22(s，1H)，8.04-7.88(m，3H)，7.58(d，J＝8.3Hz，2H)，7.24-7.10(m，2H)，6.96(t，J＝7.1Hz，1H)，6.85-6.72(m，1H)，6.58(t，J＝7.3Hz，1H)，4.99-4.72(m，4H).ESI-MS(m/s)：396.38[M+1]，394.17[M-1].4-(3-((6-fluoropyridin-3-yl)methyl)thioureido)benzoic acid (90mg, 0.30mmol) was dissolved in dry DMF (5mL) and o-phenylenediamine (32mg, 0.30mmol) was added , HBTU (114 mg, 0.30 mmol), triethylamine (41 μL, 0.30 mmol), react at room temperature for 4 hours. After the reaction, the reaction solution was poured into water (40 mL), and the crude product was obtained by filtration. The crude product was subjected to column chromatography (dichloromethane:methanol=100:3) to obtain 59 mg of a white powder with a yield of 51%. 1 H-NMR (DMSO-d 6 , 300MHz) δ: 9.95(s, 1H), 9.56(s, 1H), 8.46(s, 1H), 8.22(s, 1H), 8.04-7.88(m, 3H) , 7.58(d, J=8.3Hz, 2H), 7.24-7.10(m, 2H), 6.96(t, J=7.1Hz, 1H), 6.85-6.72(m, 1H), 6.58(t, J=7.3 Hz, 1H), 4.99-4.72 (m, 4H). ESI-MS (m/s): 396.38[M+1], 394.17[M-1].

其他A系列化合物A2-20制备方法参照实施例1。Refer to Example 1 for the preparation method of other series A compounds A2-20.

实施例2、N-(4-((2-氨基苯基)氨基甲酰基)苯基)味唑[1，2-a]吡啶-7-甲酰胺制Embodiment 2, N-(4-((2-aminophenyl) carbamoyl) phenyl) imidazo [1,2-a] pyridine-7-carboxamide 备(B7)Preparation (B7)

2.1制备中间体8：4-((咪唑并[1，2-a]吡啶-7-甲酰胺)甲基)苯甲酸甲酯2.1 Preparation of Intermediate 8: Methyl 4-((imidazo[1,2-a]pyridine-7-carboxamide)methyl)benzoate

将咪唑并[1，2-a]吡啶-6-甲酸(200mg，1.00mmol)，EDC(230mg，1.20mmol)和DMAP(146mg，1.20mmol)溶于干燥的DMF(5mL)，室温条件下搅拌0.5小时。再加入4-氨甲基苯甲酸甲酯(165mg，1.00mmol)，继续搅拌2小时。反应完后将反应液倒入十倍量的水中，有固体析出。将固体过滤，滤饼烘干后柱层析获得中间体120mg，收率是37％。1H-NMR(DMSO-d6，300MHz)δ：9.21(t，J＝5.7Hz，1H)，9.16(s，1H)，8.08(s，1H)，7.90(d，J＝8.3Hz，2H)，7.73-7.60(m，3H)，7.44(d，J＝8.4Hz，2H)，4.57(d，J＝5.8Hz，2H)，3.84(s，3H).ESI-MS(m/s)：310.51[M+1]，300.23[M-1].Dissolve imidazo[1,2-a]pyridine-6-carboxylic acid (200mg, 1.00mmol), EDC (230mg, 1.20mmol) and DMAP (146mg, 1.20mmol) in dry DMF (5mL), and stir at room temperature 0.5 hours. Additional methyl 4-aminomethylbenzoate (165 mg, 1.00 mmol) was added and stirring was continued for 2 hours. After the reaction, the reaction solution was poured into ten times the amount of water, and solids were precipitated. The solid was filtered, and the filter cake was dried and then column chromatographed to obtain 120 mg of the intermediate, with a yield of 37%. 1 H-NMR (DMSO-d 6 , 300MHz) δ: 9.21(t, J=5.7Hz, 1H), 9.16(s, 1H), 8.08(s, 1H), 7.90(d, J=8.3Hz, 2H ), 7.73-7.60(m, 3H), 7.44(d, J=8.4Hz, 2H), 4.57(d, J=5.8Hz, 2H), 3.84(s, 3H).ESI-MS(m/s) : 310.51[M+1], 300.23[M-1].

2.2制备中间体9：4-((咪唑并[1，2-a]吡啶-7-甲酰胺)甲基)苯甲酸2.2 Preparation of Intermediate 9: 4-((imidazo[1,2-a]pyridine-7-carboxamide)methyl)benzoic acid

4-((咪唑并[1，2-a]吡啶-7-甲酰胺)甲基)苯甲酸甲酯(120mg.0.387mmol)溶于THF∶MeOH∶H2O为3∶2∶1(6mL)的混合溶剂中。加入LiOH(28mg，1.16mmol)，室温条件下反应24小时。反应完后蒸干溶剂，残余物加水(10mL)，用稀盐酸调PH值到3-4，有白色固体析出，过滤后滤饼烘干，得到白色固体产物111mg，收率为97％。1H-NMR(DMSO-d6，300Hz)δ：12.92(sbr，1H)，9.22(t，J＝5.7Hz，1H)，9.17(s，1H)，8.08(s，1H)，7.91(d，J＝8.3Hz，2H)，7.72-7.60(m，3H)，7.45(d，J＝8.3Hz，2H)，4.57(d，J＝5.7Hz，2H).ESI-MS(m/s)：296.42[M+1]，294.19[M-1].Methyl 4-((imidazo[1,2-a]pyridine-7-carboxamide)methyl)benzoate (120mg.0.387mmol) was dissolved in THF:MeOH:H 2 O at a ratio of 3:2:1 (6mL ) in a mixed solvent. LiOH (28 mg, 1.16 mmol) was added and reacted at room temperature for 24 hours. After the reaction, the solvent was evaporated to dryness, water (10 mL) was added to the residue, and the pH value was adjusted to 3-4 with dilute hydrochloric acid. A white solid was precipitated. After filtration, the filter cake was dried to obtain 111 mg of a white solid product with a yield of 97%. 1 H-NMR (DMSO-d 6 , 300Hz) δ: 12.92(sbr, 1H), 9.22(t, J=5.7Hz, 1H), 9.17(s, 1H), 8.08(s, 1H), 7.91(d , J=8.3Hz, 2H), 7.72-7.60(m, 3H), 7.45(d, J=8.3Hz, 2H), 4.57(d, J=5.7Hz, 2H).ESI-MS(m/s) : 296.42[M+1], 294.19[M-1].

2.3制备目标产物B72.3 Preparation of target product B7

4-((咪唑并[1，2-a]吡啶-7-甲酰胺)甲基)苯甲酸(111mg，0.38mmol)、邻苯二胺(45mg，0.41mmol)、HBTU(157mg，0.41mmol)溶于干燥的DMF(3mL)中，滴加三乙胺(60μL，0.41mmol)，然后室温搅拌反应4小时。待反应完全后，将反应液倒入的水中(30mL)，乙酸乙酯萃取(30mL×3)，无水硫酸钠干燥。萃取液蒸干，残余物柱层析(二氯甲烷∶甲醇＝100∶3)得到白色粉末状固体56mg，收率39％。1H-NMR(DMSO-d6，300MHz)δ：9.62(s，1H)，9.22(t，J＝6.1Hz，1H)，9.17(s，1H)，8.08(s，1H)，7.94(d，J＝8.4Hz，2H)，7.77-7.57(m，3H)，7.45(d，J＝8.2Hz，2H)，7.15(d，J＝7.6Hz，1H)，6.95(t，J＝6.9Hz，1H)，6.76(d，J＝7.1Hz，1H)，6.58(t，J＝7.4Hz，1H)，4.57(d，J＝4.6Hz，2H).13C-NMR(DMSO-d6，150MHz)δ：165.57，164.76，144.84，143.55，143.38，143.26，134.37，133.68，129.15，128.27，127.80，127.63，127.15，126.94，124.97，123.79，123.66，120.24，119.59，116.74，116.60，116.40，114.92，110.07，49.06，42.96.ESI-MS(m/s)：386.38[M+1]，384.30[M-1].4-((imidazo[1,2-a]pyridine-7-carboxamide)methyl)benzoic acid (111mg, 0.38mmol), o-phenylenediamine (45mg, 0.41mmol), HBTU (157mg, 0.41mmol) Dissolve in dry DMF (3 mL), add triethylamine (60 μL, 0.41 mmol) dropwise, and then stir the reaction at room temperature for 4 hours. After the reaction was complete, the reaction solution was poured into water (30 mL), extracted with ethyl acetate (30 mL×3), and dried over anhydrous sodium sulfate. The extract was evaporated to dryness, and the residue was subjected to column chromatography (dichloromethane:methanol=100:3) to obtain 56 mg of a white powdery solid with a yield of 39%. 1 H-NMR (DMSO-d 6 , 300MHz) δ: 9.62(s, 1H), 9.22(t, J=6.1Hz, 1H), 9.17(s, 1H), 8.08(s, 1H), 7.94(d , J=8.4Hz, 2H), 7.77-7.57(m, 3H), 7.45(d, J=8.2Hz, 2H), 7.15(d, J=7.6Hz, 1H), 6.95(t, J=6.9Hz , 1H), 6.76(d, J=7.1Hz, 1H), 6.58(t, J=7.4Hz, 1H), 4.57(d, J=4.6Hz, 2H). 13 C-NMR (DMSO-d 6 , 150MHz)δ：165.57，164.76，144.84，143.55，143.38，143.26，134.37，133.68，129.15，128.27，127.80，127.63，127.15，126.94，124.97，123.79，123.66，120.24，119.59，116.74，116.60，116.40，114.92， 110.07, 49.06, 42.96. ESI-MS (m/s): 386.38[M+1], 384.30[M-1].

其他B系列化合物B1-6制备方法参照实施例2。Refer to Example 2 for the preparation methods of other B series compounds B1-6.

实施例3、N-(2-氨基苯基)-4-((4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲Example 3, N-(2-aminophenyl)-4-((4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)methyl)benzyl 酰胺制备(C1)Amide Preparation (C1)

3.1制备中间体15：4-叠氮甲基苯甲酸甲酯3.1 Preparation of intermediate 15: methyl 4-azidomethylbenzoate

4-溴甲基苯甲酸甲酯(342mg，1.5mmol)溶于干燥DMF(5mL)，室温搅拌下缓慢加入NaN3(195mg，3mmol)。然后，将反应液置于80℃的油浴中反应8小时。反应结束后将反应液倒入水中(50mL)，乙酸乙酯萃取(50mL×3)，无水硫酸镁干燥。溶剂蒸干即获得中间体4-叠氮甲基苯甲酸甲酯310mg，收率85％。1H-NMR(DMSO-d6，300MHz)δ：7.90(d，J＝8.3Hz，2H)，7.43(d，J＝8.3Hz，2H)，3.83(s，3H)，3.77(t，J＝7.0Hz，2H)，3.20(t，J＝7.0Hz，2H).ESI-MS(m/s)：206.35[M+1].Methyl 4-bromomethylbenzoate (342 mg, 1.5 mmol) was dissolved in dry DMF (5 mL), and NaN3 (195 mg, 3 mmol) was added slowly with stirring at room temperature. Then, the reaction solution was placed in an oil bath at 80° C. for 8 hours. After the reaction, the reaction solution was poured into water (50 mL), extracted with ethyl acetate (50 mL×3), and dried over anhydrous magnesium sulfate. The solvent was evaporated to dryness to obtain 310 mg of the intermediate methyl 4-azidomethylbenzoate with a yield of 85%. 1 H-NMR (DMSO-d 6 , 300MHz) δ: 7.90(d, J=8.3Hz, 2H), 7.43(d, J=8.3Hz, 2H), 3.83(s, 3H), 3.77(t, J =7.0Hz, 2H), 3.20(t, J=7.0Hz, 2H).ESI-MS (m/s): 206.35[M+1].

3.2制备中间体16：4-((4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酸甲酯3.2 Preparation of Intermediate 16: Methyl 4-((4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)methyl)benzoate

3-乙炔基吡啶(154mg，1.5mmol)、4-叠氮甲基苯甲酸甲(287mg，1.5mmol)、Vc Na(30mg，0.15mmol)和CuSO4.5H2O(3.8mg，0.015mmol)溶于叔丁醇/水(1∶1，6mL)混合溶剂中，N2保护条件下室温反应12小时。待反应结束后，将反应液倒入水中(30mL)，有白色固体析出。将混合液过滤，得到滤饼。滤饼经柱层析(石油醚∶乙酸乙酯＝1∶2)得到白色粉末状固体360mg，收率为82％。1H-NMR(DMSO-d6，300Hz)δ：9.04(d，J＝1.7Hz，1H)，8.79(s，1H)，8.53(dd，J＝1.5，4.7Hz，1H)，8.21(tt，J＝1.8，7.9Hz，1H)，7.97(d，J＝8.2Hz，2H)，7.49-7.43(m，3H)，5.78(s，2H)，3.83(s，3H).ESI-MS(m/s)：295.35[M+1]，589.39[2M+1].3-ethynylpyridine (154mg, 1.5mmol), methyl 4-azidomethylbenzoate (287mg, 1.5mmol), Vc Na (30mg, 0.15mmol) and CuSO 4 .5H 2 O (3.8mg, 0.015mmol) Dissolve in tert-butanol/water (1:1, 6mL) mixed solvent, and react at room temperature for 12 hours under N2 protection. After the reaction was completed, the reaction solution was poured into water (30 mL), and a white solid was precipitated. The mixture was filtered to obtain a filter cake. The filter cake was subjected to column chromatography (petroleum ether: ethyl acetate = 1:2) to obtain 360 mg of a white powdery solid with a yield of 82%. 1 H-NMR (DMSO-d 6 , 300Hz) δ: 9.04(d, J=1.7Hz, 1H), 8.79(s, 1H), 8.53(dd, J=1.5, 4.7Hz, 1H), 8.21(tt , J=1.8, 7.9Hz, 1H), 7.97(d, J=8.2Hz, 2H), 7.49-7.43(m, 3H), 5.78(s, 2H), 3.83(s, 3H).ESI-MS( m/s): 295.35[M+1], 589.39[2M+1].

3.3制备中间体17：4-((4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酸3.3 Preparation of intermediate 17: 4-((4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)methyl)benzoic acid

将中间体16(303mg，1.03mmol)溶于THF∶MeOH∶H2O为3∶2∶1的混合溶剂中(12mL)。加入LiOH(74mg，3.09mmol)，室温条件下反应24小时。反应完后蒸干溶剂，加入15mL的水溶液，用稀盐酸调PH值至3-4，有白色固体析出，过滤后滤饼烘干，即为产物260mg，收率为90％。1H-NMR(DMSO-d6，300MHz)δ：13.05(s，1H)，9.06(s，1H)，8.80(s，1H)，8.55-8.53(m，1H)，8.23(d，J＝8.0Hz，1H)，7.96(d，J＝8.4Hz，2H)，7.50-7.43(m，3H)，5.79(s，2H).ESI-MS(m/s)：295.35[M+1].Intermediate 16 (303 mg, 1.03 mmol) was dissolved in a THF:MeOH:H 2 O 3:2:1 mixed solvent (12 mL). LiOH (74mg, 3.09mmol) was added and reacted at room temperature for 24 hours. After the reaction, the solvent was evaporated to dryness, 15 mL of aqueous solution was added, and the pH value was adjusted to 3-4 with dilute hydrochloric acid. A white solid was precipitated. After filtering, the filter cake was dried to obtain 260 mg of the product with a yield of 90%. 1 H-NMR (DMSO-d6, 300MHz) δ: 13.05(s, 1H), 9.06(s, 1H), 8.80(s, 1H), 8.55-8.53(m, 1H), 8.23(d, J=8.0 Hz, 1H), 7.96(d, J=8.4Hz, 2H), 7.50-7.43(m, 3H), 5.79(s, 2H). ESI-MS(m/s): 295.35[M+1].

3.4制备目标产物C13.4 Preparation of target product C1

4-((4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)甲基)苯甲酸(250mg，0.89mmol)、邻苯二胺(106mg，0.98mmol)、HBTU(372mg，0.98mmol)溶于DMF(5mL)中，滴加三乙胺(136μL，0.98mmol)，室温搅拌反应4小时。待反应完全后，将反应液倒入水中(40mL)，乙酸乙酯萃取(40mL×3)，合并有机层，无水硫酸钠干燥，减压蒸干溶液，残余物经柱层析(二氯甲烷∶甲醇＝100∶2)得到淡黄色粉末固体88mg，收率27％。1H-NMR(DMSO-d6，300Hz)δ：9.64(s，1H)，9.06-9.05(d，J＝1.9Hz，1H)，8.79(s，1H)，8.53(dd，J＝1.4，4.7Hz，1H)，8.21(dt，J＝1.7，7.9Hz，1H)，7.98(d，J＝8.1Hz，2H)，7.49-7.45(m，3H)，7.14(d，J＝7.8Hz，1H)，6.98-6.92(m，1H)，6.77-6.74(m，1H)，6.58(t，J＝7.6Hz，1H)，5.77(s，2H)，4.88(s，2H).ESI-MS(m/s)：371.29[M+1].4-((4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)methyl)benzoic acid (250mg, 0.89mmol), o-phenylenediamine (106mg, 0.98mmol ), HBTU (372 mg, 0.98 mmol) were dissolved in DMF (5 mL), triethylamine (136 μL, 0.98 mmol) was added dropwise, and the reaction was stirred at room temperature for 4 hours. After the reaction is complete, the reaction solution is poured into water (40mL), extracted with ethyl acetate (40mL×3), the organic layers are combined, dried over anhydrous sodium sulfate, the solution is evaporated to dryness under reduced pressure, and the residue is subjected to column chromatography (dichloro Methane:methanol=100:2) to obtain 88 mg of light yellow powder solid, yield 27%. 1 H-NMR (DMSO-d 6 , 300Hz) δ: 9.64 (s, 1H), 9.06-9.05 (d, J=1.9Hz, 1H), 8.79 (s, 1H), 8.53 (dd, J=1.4, 4.7Hz, 1H), 8.21(dt, J=1.7, 7.9Hz, 1H), 7.98(d, J=8.1Hz, 2H), 7.49-7.45(m, 3H), 7.14(d, J=7.8Hz, 1H), 6.98-6.92(m, 1H), 6.77-6.74(m, 1H), 6.58(t, J=7.6Hz, 1H), 5.77(s, 2H), 4.88(s, 2H).ESI-MS (m/s): 371.29[M+1].

其他C系列化合物C2-14制备方法参照实施例3。Refer to Example 3 for the preparation method of other C series compounds C2-14.

实施例4、N-(2-氨基苯基)-8-(4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)辛酰胺(D1)Example 4, N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)octylamide (D1)

4.1制备中间体20：N-(2-氨基苯基)-8-叠氮基辛酰胺4.1 Preparation of intermediate 20: N-(2-aminophenyl)-8-azidooctylamide

8-叠氮基辛酸(0.43g，2.32mmol)与邻苯二胺(0.28g，2.55mmol)溶于5mL DMF中，依次加入HBTU(1.06g，2.79mmol)与三乙胺1.30mL，室温下搅拌30分钟，待反应完全后，将反应液倾入水中(50mL)，用乙酸乙酯(50mL×3)萃取，合并有机层，干燥，减压旋干，残余物经柱层析(二氯甲烷∶甲醇＝100∶1)得白色固体0.07g，收率92.1％。1H-NMR(DMSO-d6，600MHz)δ：1.29-1.33(m，6H)，1.50-1.55(m，2H)，1.55-1.59(m，2H)，2.29(t，J＝7.78Hz，2H)，3.31(t，J＝6.91Hz，2H)，4.80(s，2H)，6.50-6.55(m，1H)，6.70(dd，J＝7.96Hz，1.38Hz，1H)，6.85-6.89(m，1H)，7.13(dd，J＝7.83Hz，1.51Hz，1H)，9.08(s，1H).MS(ESI，positive)m/z：276.17[M+H].8-Azidooctanoic acid (0.43g, 2.32mmol) and o-phenylenediamine (0.28g, 2.55mmol) were dissolved in 5mL of DMF, HBTU (1.06g, 2.79mmol) and 1.30mL of triethylamine were added successively, and the Stir for 30 minutes. After the reaction is complete, pour the reaction solution into water (50mL), extract with ethyl acetate (50mL×3), combine the organic layers, dry, and spin dry under reduced pressure. The residue is subjected to column chromatography (dichloro Methane:methanol=100:1) to obtain 0.07 g of white solid, yield 92.1%. 1 H-NMR (DMSO-d 6 , 600MHz) δ: 1.29-1.33(m, 6H), 1.50-1.55(m, 2H), 1.55-1.59(m, 2H), 2.29(t, J=7.78Hz, 2H), 3.31(t, J=6.91Hz, 2H), 4.80(s, 2H), 6.50-6.55(m, 1H), 6.70(dd, J=7.96Hz, 1.38Hz, 1H), 6.85-6.89( m, 1H), 7.13(dd, J=7.83Hz, 1.51Hz, 1H), 9.08(s, 1H). MS(ESI, positive) m/z: 276.17[M+H].

4.2制备目标产物D14.2 Preparation of target product D1

化合物20(0.2g，0.726mmol)溶于叔丁醇和水(5∶1)的混合溶剂中(10mL)，N2保护，依次注入CuSO4水溶液(1.45mL)和VcNa水溶液(7.26mL)，室温下搅拌过夜。待反应完全后，将反应液减压旋干，剩余水层用乙酸乙酯萃取(50mL×3)，合并有机层，溶液蒸干，残余物经柱层析(二氯甲烷∶甲醇＝100∶2)得到白色固体0.16g，收率62％。1H-NMR(DMSO-d6，600MHz)δ：1.28-1.35(m，6H)，1.55-1.59(m，2H)，1.85-1.88(m，2H)，2.29(t，J＝7.78Hz，2H)，4.41(t，J＝7.08Hz，2H)，4.78(s，2H)，6.49-6.53(m，1H)，6.69(dd，J＝7.91Hz，1.23Hz，1H)，6.85-6.88(m，1H)，7.12(d，J＝6.74Hz，1H)，7.46(dd，J＝7.97Hz，4.81Hz，1H)，8.18-8.20(m，1H)，8.52(dd，J＝4.91Hz，1.35Hz，1H)，8.70(s，1H)，9.03(d，J＝1.44Hz，1H)，9.06(s，1H).MS(ESI，positive)m/z：379.22[M+H].Compound 20 (0.2g, 0.726mmol) was dissolved in a mixed solvent of tert-butanol and water (5:1) (10mL), protected by N 2 , injected into CuSO 4 aqueous solution (1.45mL) and VcNa aqueous solution (7.26mL) successively, at room temperature Stir overnight. After the reaction was complete, the reaction solution was spin-dried under reduced pressure, the remaining aqueous layer was extracted with ethyl acetate (50mL×3), the organic layers were combined, the solution was evaporated to dryness, and the residue was subjected to column chromatography (dichloromethane:methanol=100: 2) Obtain 0.16 g of white solid, yield 62%. 1 H-NMR (DMSO-d 6 , 600MHz) δ: 1.28-1.35(m, 6H), 1.55-1.59(m, 2H), 1.85-1.88(m, 2H), 2.29(t, J=7.78Hz, 2H), 4.41(t, J=7.08Hz, 2H), 4.78(s, 2H), 6.49-6.53(m, 1H), 6.69(dd, J=7.91Hz, 1.23Hz, 1H), 6.85-6.88( m, 1H), 7.12(d, J=6.74Hz, 1H), 7.46(dd, J=7.97Hz, 4.81Hz, 1H), 8.18-8.20(m, 1H), 8.52(dd, J=4.91Hz, 1.35Hz, 1H), 8.70(s, 1H), 9.03(d, J=1.44Hz, 1H), 9.06(s, 1H). MS(ESI, positive) m/z: 379.22[M+H].

其他D系列化合物D2-5制备方法参照实施例4。Refer to Example 4 for the preparation method of other D series compounds D2-5.

实施例5、N-(2-氨基苯基)-(4-(4-(4-((吡啶-3-撑甲基)氨甲酰基)苯基)-1H-1，Example 5, N-(2-aminophenyl)-(4-(4-(4-((pyridine-3-enylmethyl)carbamoyl)phenyl)-1H-1, 2，3-三唑-1-基)甲基)苯甲酰胺盐酸盐制备(化合物C14盐酸盐)Preparation of 2,3-triazol-1-yl)methyl)benzamide hydrochloride (compound C14 hydrochloride)

将0.1g的N-(2-氨基苯基)-(4-(4-(4-((吡啶-3-撑甲基)氨甲酰基)苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺溶于5mL的热乙醇中，持续通入干燥氯化氢气体。反应搅拌2小时，有白色固体析出，冷却后抽滤、洗涤、干燥得到N-(2-氨基苯基)-(4-(4-(4-((吡啶-3-撑甲基)氨甲酰基)苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺盐酸盐0.05g。0.1 g of N-(2-aminophenyl)-(4-(4-(4-((pyridine-3-enyl)carbamoyl)phenyl)-1H-1,2,3-tri Azol-1-yl)methyl)benzamide was dissolved in 5mL of hot ethanol, and dry hydrogen chloride gas was continuously passed through. The reaction was stirred for 2 hours, and a white solid was precipitated. After cooling, it was filtered with suction, washed, and dried to obtain N-(2-aminophenyl)-(4-(4-(4-((pyridine-3-ylidenemethyl)aminomethyl) Acyl)phenyl)-1H-1,2,3-triazol-1-yl)methyl)benzamide hydrochloride 0.05g.

实施例6、N-(2-氨基苯基)-4-(3-(吡啶-3-甲撑基)脲基)苯甲酰胺琥珀酸盐制备Example 6, Preparation of N-(2-aminophenyl)-4-(3-(pyridine-3-methylene)ureido)benzamide succinate (化合物A2琥珀酸盐)(Compound A2 Succinate)

将0.5g的N-(2-氨基苯基)-4-(3-(吡啶-3-甲撑基)脲基)苯甲酰胺琥珀酸盐溶于10mL的热乙醇中，加入热的琥珀酸乙醇溶液，室温反应3小时，反应完后抽滤、洗涤、干燥得到N-(2-氨基苯基)-4-(3-(吡啶-3-甲撑基)脲基)苯甲酰胺琥珀酸盐0.1g。Dissolve 0.5 g of N-(2-aminophenyl)-4-(3-(pyridin-3-methylene)ureido)benzamide succinate in 10 mL of hot ethanol, add hot succinic acid Ethanol solution, reacted at room temperature for 3 hours, after the reaction was completed, suction filtered, washed, and dried to obtain N-(2-aminophenyl)-4-(3-(pyridine-3-methylene)ureido)benzamide succinic acid Salt 0.1g.

实施例7、N-(2-氨基苯基)-(4-(4-(4-((吡啶-3-撑甲基)氨甲酰基)苯基)-1H-1，Example 7, N-(2-aminophenyl)-(4-(4-(4-((pyridine-3-enylmethyl)carbamoyl)phenyl)-1H-1, 2，3-三唑-1-基)甲基)苯甲酰胺马来酸盐制备(化合物C14马来酸盐)Preparation of 2,3-triazol-1-yl)methyl)benzamide maleate (compound C14 maleate)

在50mL圆底烧瓶中，加入0.3gN-(2-氨基苯基)-(4-(4-(4-((吡啶-3-撑甲基)氨甲酰基)苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺，乙醇溶解，加入热的马来酸乙醇溶液，室温反应3小时，反应完后抽滤、洗涤、干燥得到N-(2-氨基苯基)-(4-(4-(4-((吡啶-3-撑甲基)氨甲酰基)苯基)-1H-1，2，3-三唑-1-基)甲基)苯甲酰胺马来酸盐0.15g。In a 50 mL round bottom flask, add 0.3 g N-(2-aminophenyl)-(4-(4-(4-((pyridin-3-ylidenemethyl)carbamoyl)phenyl)-1H-1, 2,3-triazol-1-yl)methyl)benzamide, dissolved in ethanol, added hot maleic acid ethanol solution, reacted at room temperature for 3 hours, after the reaction was completed, suction filtered, washed, and dried to obtain N-(2- Aminophenyl)-(4-(4-(4-((pyridine-3-enylmethyl)carbamoyl)phenyl)-1H-1,2,3-triazol-1-yl)methyl) Benzamide maleate 0.15g.

实施例8、N-(2-氨基苯基)-4-(2-(4-(吡啶-3-基))-1H-1，2，3-三唑-1-基)苯甲酰Example 8, N-(2-aminophenyl)-4-(2-(4-(pyridin-3-yl))-1H-1,2,3-triazol-1-yl)benzoyl 胺苹果酸盐制备(化合物C1苹果酸盐)Amine malate preparation (compound C1 malate)

在50mL圆底烧瓶中，加入0.5gN-(2-氨基苯基)-4-(2-(4-(吡啶-3-基))-1H-1，2，3-三唑-1-基)苯甲酰胺，乙醇溶解，加入热的苹果酸乙醇溶液，室温反应3小时，反应完后抽滤、洗涤、干燥得到N-(2-氨基苯基)-4-(2-(4-(吡啶-3-基))-1H-1，2，3-三唑-1-基)苯甲酰胺苹果酸盐0.09g。In a 50 mL round bottom flask, add 0.5 g of N-(2-aminophenyl)-4-(2-(4-(pyridin-3-yl))-1H-1,2,3-triazol-1-yl ) benzamide, dissolved in ethanol, added hot malic acid ethanol solution, reacted at room temperature for 3 hours, suction filtered, washed, and dried to obtain N-(2-aminophenyl)-4-(2-(4-( Pyridin-3-yl))-1H-1,2,3-triazol-1-yl)benzamide malate 0.09 g.

实施例9、N-(2-氨基苯基)-8-(4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)辛酰胺酒石Example 9, N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)octylamide tartar 酸盐制备(化合物D1酒石酸盐)Salt preparation (compound D1 tartrate)

在50mL圆底烧瓶中，加入0.5gN-(2-氨基苯基)-8-(4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)辛酰胺，乙醇溶解，加入热的酒石酸乙醇溶液，室温反应3小时，反应完后抽滤、洗涤、干燥得到N-(2-氨基苯基)-8-(4-(吡啶-3-基)-1H-1，2，3-三唑-1-基)辛酰胺酒石酸盐0.23g。In a 50mL round bottom flask, add 0.5g N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H-1,2,3-triazol-1-yl)octylamide, Dissolved in ethanol, added hot ethanol tartaric acid solution, reacted at room temperature for 3 hours, after the reaction was completed, suction filtered, washed, and dried to obtain N-(2-aminophenyl)-8-(4-(pyridin-3-yl)-1H- 0.23 g of 1,2,3-triazol-1-yl) octanamide tartrate.

实施例10、目标化合物酶抑制活性和体外抗肿瘤活性Example 10, target compound enzyme inhibitory activity and in vitro antitumor activity

(1)目标化合物NAMPT酶抑制测试(1) Target compound NAMPT enzyme inhibition test

以下的酶是指烟酰胺磷酸核糖转移酶，可购自康肽生物科技有限公司，也可自行制备。The following enzyme refers to nicotinamide phosphoribosyltransferase, which can be purchased from Kangtai Biotechnology Co., Ltd. or prepared by itself.

1)酶的制备：1) Preparation of enzyme:

将转化有重组质粒NAMPT-pET28a+的BL21(DE3)plysS细胞接种于2×YT培养基(37μg/mL氯霉素和100μg/mL卡那霉素)中，37℃振摇过夜，收集菌体后用20倍于原体积的新鲜培养基重悬，37℃培养至OD600约0.6-0.8，在0.5mM IPTG、28℃条件下诱导8小时。离心收集菌体，并重悬于裂解缓冲液(20mM Tris-HCl pH 8.0，300mM NaCl)中，加入0.1％Triton X-100，1％PMSF，200W超声裂解细胞，超声1s间隙9s，共进行30min。将裂解液于12000rpm、4℃离心50min吸取上清液。该上清液与Ni-NTA柱(购自QIAGEN公司)在冰上振摇孵育2小时，再依次用结合缓冲液(5mM咪唑，0.5M NaCl，20mM Tris-Hcl，pH＝7.5)、缓冲液(10mM/20mM/40mM/60mM咪唑，0.5M NaCl，20mM Tris-HCl，pH＝7.5)依次洗去杂蛋白，最后用洗脱缓冲液(200mM咪唑，0.5M NaCl，20mM Tris-HCl，PH＝7.5)洗脱目的蛋白，并进行SDS-PAGE检测，用BCA方法测定蛋白浓度。Inoculate BL21(DE3)plysS cells transformed with the recombinant plasmid NAMPT-pET28a + in 2×YT medium (37 μg/mL chloramphenicol and 100 μg/mL kanamycin), shake overnight at 37°C, and collect the cells Then resuspend with 20 times the original volume of fresh medium, culture at 37°C to an OD600 of about 0.6-0.8, and induce for 8 hours at 0.5mM IPTG at 28°C. The cells were collected by centrifugation, resuspended in lysis buffer (20mM Tris-HCl pH 8.0, 300mM NaCl), added 0.1% Triton X-100, 1% PMSF, 200W ultrasonic lysed cells, ultrasonic 1s interval 9s, a total of 30min. The lysate was centrifuged at 12000 rpm and 4°C for 50 min to absorb the supernatant. The supernatant was incubated with a Ni-NTA column (purchased from QIAGEN) for 2 hours with shaking on ice, and then sequentially washed with binding buffer (5mM imidazole, 0.5M NaCl, 20mM Tris-Hcl, pH=7.5), buffer (10mM/20mM/40mM/60mM imidazole, 0.5M NaCl, 20mM Tris-HCl, pH=7.5) to wash off the foreign protein in sequence, and finally use the elution buffer (200mM imidazole, 0.5M NaCl, 20mM Tris-HCl, pH= 7.5) The target protein is eluted and detected by SDS-PAGE, and the protein concentration is determined by the BCA method.

2)酶反应体系为25μL，其中各种组分的终浓度为：50mM Tris-HCl(pH 7.5)、0.02％BSA，12mM MgCl2、2mM ATP、0.4mM PRPP、2mM DTT、2μg/mL NAMPT、0.2μM NAM、DMSO和倍比稀释的本发明的化合物。先将0.5μL本发明的化合物的不同浓度溶液加于96孔板，再加入20μL酶反应混合溶液(除底物之外的酶反应组分)，于室温孵育5分钟后，加入4.5μL底物NAM溶液以启动反应，37℃反应15分钟后，于95℃加热1分钟终止酶反应。2) The enzyme reaction system is 25 μL, and the final concentrations of various components are: 50mM Tris-HCl (pH 7.5), 0.02% BSA, 12mM MgCl 2 , 2mM ATP, 0.4mM PRPP, 2mM DTT, 2μg/mL NAMPT, 0.2 μM NAM, DMSO and compound of the invention in serial dilutions. First add 0.5 μL of different concentration solutions of the compound of the present invention to a 96-well plate, then add 20 μL of enzyme reaction mixture solution (enzyme reaction components other than the substrate), incubate at room temperature for 5 minutes, then add 4.5 μL of substrate NAM solution was used to initiate the reaction, and after 15 minutes of reaction at 37°C, the enzyme reaction was terminated by heating at 95°C for 1 minute.

3)将反应液在冰上冷却后，依次加入10μL 20％苯乙酮和2M KOH，涡旋混合仪上混匀后于0℃作用2分钟，加入45μL 88％甲酸，37℃孵育10分钟。3) After the reaction solution was cooled on ice, 10 μL of 20% acetophenone and 2M KOH were added sequentially, mixed on a vortex mixer and then reacted at 0° C. for 2 minutes, then added 45 μL of 88% formic acid, and incubated at 37° C. for 10 minutes.

4)使用酶标仪测定激发波长382nm、发射波长445nm处的荧光值。4) Use a microplate reader to measure the fluorescence value at the excitation wavelength of 382nm and the emission wavelength of 445nm.

5)根据公式：E＝R/(1+(C/IC50)s)+B(其中E为酶活性，C为化合物浓度，R、IC50、S、B为待拟合的参数)，在origin软件中将酶活性对化合物浓度的曲线进行拟合，求出化合物的IC50。5) According to the formula: E=R/(1+(C/IC 50 ) s )+B (where E is the enzyme activity, C is the compound concentration, R, IC 50 , S, and B are the parameters to be fitted), The curve of enzyme activity versus compound concentration was fitted in the origin software, and the IC 50 of the compound was calculated.

(2)目标化合物HDAC1酶抑制测试(2) Target compound HDAC1 enzyme inhibition test

1)实验材料：1) Experimental materials:

HDAC1酶，缓冲液(137mM氯化钠，2.7mM氯化钾，1mM氯化镁，0.1mg/mL BSA，PH＝8的Tris-HCl 25mM)，HDAC底物3，胰蛋白酶，96孔黑色板。HDAC1 enzyme, buffer (137 mM sodium chloride, 2.7 mM potassium chloride, 1 mM magnesium chloride, 0.1 mg/mL BSA, Tris-HCl 25 mM pH=8), HDAC substrate 3, trypsin, 96-well black plate.

2)实验方法：2) Experimental method:

(a)96孔黑色板平衡至室温。(a) The 96-well black plate is equilibrated to room temperature.

(b)用含有10％的DMSO的缓冲液稀释待测化合物，化合物的浓度依次为100μM，30μM，10μM，3μM，1μM，0.3μM，0.1μM，0.03μM，0.01μM，0.003μM。(b) Dilute the test compound with a buffer solution containing 10% DMSO, the concentration of the compound is 100 μM, 30 μM, 10 μM, 3 μM, 1 μM, 0.3 μM, 0.1 μM, 0.03 μM, 0.01 μM, 0.003 μM.

(c)将11μL的HDAC1加入到400μL的缓冲液中，摇匀；(c) Add 11 μL of HDAC1 to 400 μL of buffer and shake well;

(d)向96孔板上第2-11孔加入35μL刚刚配好的含有HDAC1酶的缓冲液，并依次加入5μL稀释好的不同浓度的化合物到对应的反应孔中，对于阴性对照(第一个孔)和空白对照孔(第十一个孔)，分别加入40μL和5μL缓冲液。(d) Add 35 μL of the newly prepared buffer solution containing HDAC1 enzyme to the 2-11 wells of the 96-well plate, and sequentially add 5 μL of diluted compounds of different concentrations to the corresponding reaction wells, for the negative control (first wells) and blank control wells (the eleventh well), add 40 μL and 5 μL buffer, respectively.

(e)向所有反应孔中加入100μM的HDAC底物35μL和0.5mg/mL的胰蛋白酶5μL，37度孵化30min后读数。(e) Add 35 μL of 100 μM HDAC substrate and 5 μL of 0.5 mg/mL trypsin to all reaction wells, incubate at 37 degrees for 30 min and read.

(f)依据公式计算抑制率：抑制率＝(100％活性孔-样品孔)/100％活性孔*100，在orgin软件中将酶活性对化合物浓度的曲线进行拟合，求出化合物的IC50值。(f) Calculate the inhibition rate according to the formula: inhibition rate=(100% active well-sample well)/100% active well*100, in the orgin software, the curve of the enzyme activity to the compound concentration is fitted to obtain the IC of the compound 50 value.

(3)目标化合物体外抗肿瘤活性测试(3) In vitro anti-tumor activity test of the target compound

1)样品配制：用DMSO(Merck)溶解成后，加入PBS(-)配成1000μM的溶液或均匀的混悬液，然后用含DMSO的PBS(-)稀释。样品终浓度100、10、1、0.1、0.01、0.001μM。1) Sample preparation: After dissolving in DMSO (Merck), add PBS (-) to make a 1000 μM solution or a homogeneous suspension, and then dilute with DMSO-containing PBS (-). The final concentrations of samples were 100, 10, 1, 0.1, 0.01, 0.001 μM.

2)细胞株2) cell line

HCT116(人肠癌细胞)、MDA-MB-231(人乳腺癌细胞)、HepG2(人肝癌细胞)，均由本实验室冻存和传代。HCT116 (human intestinal cancer cells), MDA-MB-231 (human breast cancer cells), HepG2 (human liver cancer cells), were all cryopreserved and passaged by our laboratory.

3)培养液3) Culture medium

DMEM+10％FBS+双抗DMEM+10%FBS+double antibody

4)试验方法4) Test method

MTT法。96孔板每孔加入浓度为4-5×104个/mL的细胞悬液100μl，置37℃，5％CO2培养箱内。24小时后，加入样品液，10μL/孔，设三复孔，37℃，5％CO2作用72小时。每孔加入5mg/mL的MTT溶液20μL，作用4小时后加入溶解液，100μL/孔，置培养箱内，溶解后用全波长多功能酶标仪测570nm OD值MTT method. Add 100 μl of cell suspension with a concentration of 4-5×10 4 cells/mL to each well of a 96-well plate, and place in a 37° C., 5% CO 2 incubator. After 24 hours, add sample solution, 10 μL/well, set up triplicate wells, 37 ° C, 5% CO 2 for 72 hours. Add 20 μL of 5 mg/mL MTT solution to each well, add the solution after 4 hours of action, 100 μL/well, put it in the incubator, measure the 570nm OD value with a full-wavelength multi-functional microplate reader after dissolution

(4)目标化合物酶抑制活性和体外抗肿瘤活性测试结果(表2)(4) Test results of target compound enzyme inhibitory activity and in vitro antitumor activity (Table 2)

表2.目标化合物酶抑制活性和体外活性结果(μM)Table 2. Enzyme inhibitory activity and in vitro activity results of target compounds (μM)

药理实验表明本发明所诉的化合物或其盐，对与肿瘤密切的相关组蛋白去乙酰化酶1亚型(HDAC1)的具有强抑制活性。Pharmacological experiments show that the compound or salt thereof of the present invention has a strong inhibitory activity on histone deacetylase 1 subtype (HDAC1) closely related to tumors.

药理实验表明本发明所诉的化合物或其盐，对和肿瘤相关的烟酰胺磷酸核糖转移酶(NAMPT)具有强抑制活性。Pharmacological experiments show that the compound or salt thereof of the present invention has strong inhibitory activity on tumor-associated nicotinamide phosphoribosyltransferase (NAMPT).

药理实验表明部分本发明所诉的化合物或其盐，对HDAC1和NAMPT均有较好的抑制活性。尤其是化合物C14对HDAC1的抑制活性为55nM，对NAMPT的抑制活性为31nM，对双靶点的抑制活性达到较好的平衡；化合物D1对HDAC1的抑制活性是15nM，对NAMPT的抑制活性为18nM，表现出较好的体外活性。化合物D2体外对HDAC1和NAMPT的抑制活性分别31nM和36nM，体外活性较好。化合物D3体外对HDACl和NAMPT的抑制活性分别40nM和41nM，体外活性较好。Pharmacological experiments show that some of the compounds or their salts described in the present invention have good inhibitory activity on HDAC1 and NAMPT. In particular, the inhibitory activity of compound C14 on HDAC1 was 55nM, and the inhibitory activity on NAMPT was 31nM, and the inhibitory activity on dual targets reached a better balance; the inhibitory activity of compound D1 on HDAC1 was 15nM, and the inhibitory activity on NAMPT was 18nM , showing good in vitro activity. The inhibitory activity of compound D2 on HDAC1 and NAMPT in vitro was 31 nM and 36 nM, respectively, and the activity in vitro was relatively good. The inhibitory activity of compound D3 on HDACl and NAMPT in vitro was 40nM and 41nM respectively, and the activity in vitro was better.

体外抗肿瘤活性测试采用MTT法，大部分化合物对肠癌(HCT116)、乳腺癌(MDA-MB-231)和肝癌(HepG2)肿瘤株均表现出较强的生长抑制活性。A2、A4、A5、A8、A13-18、A20、B1-3、B5、B7、C1-5、C7-10、C14、D3对肠癌活性优于阳性药物CI-994。A1-3、A5、A8、A12-18、A20、B1-5、B7、C1-10、C12-14、D1-3、D5对肝癌活性优于阳性药CI-994。In vitro anti-tumor activity was tested by MTT method, and most of the compounds showed strong growth inhibitory activity against colon cancer (HCT116), breast cancer (MDA-MB-231) and liver cancer (HepG2) tumor lines. A2, A4, A5, A8, A13-18, A20, B1-3, B5, B7, C1-5, C7-10, C14, D3 are more active than the positive drug CI-994 in colon cancer. A1-3, A5, A8, A12-18, A20, B1-5, B7, C1-10, C12-14, D1-3, D5 were more active against liver cancer than positive drug CI-994.

实施例11、目标化合物HDAC选择性测试Embodiment 11, target compound HDAC selectivity test

药理实验结果(表3)表明化合物C14对HDAC1和HDAC2具有选择性抑制作用。The results of pharmacological experiments (Table 3) show that compound C14 has a selective inhibitory effect on HDAC1 and HDAC2.

表3.化合物C14对HDAC各亚型抑制结果Table 3. Inhibition results of compound C14 on HDAC subtypes

实施例12、目标化合物体内抗肿瘤效果Embodiment 12, antitumor effect of target compound in vivo

根据体外抗肿瘤实验结果及化合物的结构特点，我们选择人结肠癌HCT116作为裸鼠移植瘤模型，以化合物C14作为研究对象，SAHA和FK866作为阳性对照药。According to the results of in vitro anti-tumor experiments and the structural characteristics of the compound, we selected human colon cancer HCT116 as the xenograft tumor model in nude mice, compound C14 as the research object, and SAHA and FK866 as positive control drugs.

设计动物实验的给药剂量为25mg/kg，一天两次，连续腹腔注射21天。结果表明裸鼠生存状态良好，未观察到明显的小鼠体重明显变化。化合物对肿瘤生长抑制率为68.88％(图1，表4)，显著高于阳性药SAHA(33.05％)和FK866(39.35％)。由此可见，HDAC/NAMPT双靶点抑制剂较单靶点抑制剂具有明显的优势，表现出高效低毒的优点，值得进一步研究。The dosage of the designed animal experiment is 25mg/kg, twice a day, for 21 consecutive days by intraperitoneal injection. The results showed that the nude mice lived in good condition, and no significant changes in the body weight of the mice were observed. The inhibitory rate of the compound on tumor growth was 68.88% (Fig. 1, Table 4), significantly higher than the positive drugs SAHA (33.05%) and FK866 (39.35%). It can be seen that HDAC/NAMPT dual-target inhibitors have obvious advantages over single-target inhibitors, showing the advantages of high efficiency and low toxicity, and are worthy of further research.

表4 目标化合物对人体肠癌HCT116裸鼠移植瘤的疗效Table 4 Efficacy of target compounds on human intestinal cancer HCT116 transplanted in nude mice

与Control比较：*P＜0.05，**P＜0.01。Compared with Control: *P<0.05, **P<0.01.

以上对本发明的具体实施例进行了描述。需要理解的是，本发明并不局限于上述特定实施方式，本领域技术人员可以在权利要求的范围内做出各种变形或修改，这并不影响本发明的实质内容。Specific embodiments of the present invention have been described above. It should be understood that the present invention is not limited to the specific embodiments described above, and those skilled in the art may make various changes or modifications within the scope of the claims, which do not affect the essence of the present invention.

Claims (10)

1. a kind of amide derivatives and its pharmaceutically acceptable salt, the amide derivatives general structure such as formula (I) institute Show：
Wherein, A groups are selected from any one in (1)~(4)：
(1) phenyl, substituted-phenyl,
(2) pyridine radicals, substituted pyridinyl,
(3) quinary heterocyclic radical, substituted five-membered heterocyclic radical,
(4) five yuan or the parallel base and bicyclic heterocycles base of hexatomic ring composition,
The substitution base on substitution base, substituted pyridinyl on the substituted-phenyl phenyl ring, the substitution base on substituted five-membered heterocycle are equal For monosubstituted or polysubstituted, and be respectively selected from it is following any one or a few：Halogen atom, the straight chained alkyl of 1-6 carbon atom, 3-6 the branched alkyl of carbon atom, hydroxyl, cyano group, cyano methyl, amino, nitro, trifluoromethyl, trifluoromethoxy, methoxy Base, ethyoxyl, propoxyl group, isopropoxy, butoxy, acetyl group；
E groups are selected from the one kind in following group：
X group is selected from the one kind in following group：
Wherein, R1 is selected from following any One or more：Hydrogen atom, halogen atom, the straight chained alkyl of 1-6 carbon atom, the branched alkyl of 3-6 carbon atom, hydroxyl, Cyano group, cyano methyl, amino, nitro, trifluoromethyl, trifluoromethoxy, methoxyl group, ethyoxyl, propoxyl group, isopropoxy, fourth Epoxide, acetyl group；
Y group is carbon atom or nitrogen-atoms；
Z group is selected from hydrogen atom, halogen atom, phenyl, hexa-member heterocycle base or quinary heterocyclic radical.
2. amide derivatives according to claim 1 and its pharmaceutically acceptable salt, it is characterised in that described five yuan Or the parallel base and bicyclic heterocycles base of hexatomic ring composition, comprising：
3. amide derivatives according to claim 1 and its pharmaceutically acceptable salt, it is characterised in that described medicine Acceptable salt is hydrochloride, hydrobromate, sulfate, acetate, lactate, tartrate, tannate, citric acid on Salt, trifluoroacetate, malate, maleate, succinate, tosilate or mesylate.
4. amide derivatives according to claim 1 and its pharmaceutically acceptable salt, it is characterised in that described medicine Acceptable salt on, without the crystallization water, or the crystallization water containing one or more molecules.
5. amide derivatives according to claim 1 and its pharmaceutically acceptable salt, it is characterised in that described medicine The crystallization water of the acceptable salt containing 0.5-3.0 molecules on.
6. amide derivatives according to claim 1 and its pharmaceutically acceptable salt, it is characterised in that the acid amides Analog derivative is：
N- (2- aminophenyls) -4- (3- (6- fluorine pyridin-3-yl) methyl) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (pyridin-3-yl methyl) urea groups) benzamide,
N- (2- aminophenyls) -4- (3- (pyridin-3-yl) acrylamido) benzamide,
N- (2- aminophenyls) -4- (3- (6- chloropyridine -3- bases) methyl) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (6- chloropyridine -3- bases) methyl) urea groups) benzamide,
N- (2- aminophenyls) -4- (3- (6- fluorine pyridin-3-yl) methyl) urea groups) benzamide,
N- (2- aminophenyls) -4- (3- (6- aminopyridine -3- bases) methyl) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (6- aminopyridine -3- bases) methyl) urea groups) benzamide,
The tert-butyl group (5- ((3- (4- ((2- aminophenyls) carbamyl) phenyl) urea groups) methyl) pyridine -2- bases) carbamic acid Ester,
N- (2- aminophenyls) -4- (3- (5- fluorine pyridin-3-yl) methyl) urea groups) benzamide,
N- (2- aminophenyls) -4- (3- (6- picoline -3- bases) methyl) urea groups) benzamide,
N- (2- aminophenyls) -4- (3- (4- luorobenzyls) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (3- luorobenzyls) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (2- luorobenzyls) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (3- chlorobenzyls) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (3- bromobenzyls) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (benzo [d] [1,3] dioxole -5- ylmethyls) ghiourea group) benzamide,
N- (2- aminophenyls) -4- (3- (3- aminobenzyls) ghiourea group) benzamide,
4- (3- ((1H- benzos [d] imidazoles -2- bases) methyl) ghiourea group)-N- (2- aminophenyls) benzamide,
N- (2- aminophenyls) -4- (2,3- bis- chloro- 3- (pyridin-3-yl) acrylamido) benzamide,
N- (4- ((2- aminophenyls) carbamoyl) benzyl) thieno [3,2-c] pyridine-2-carboxamide,
N- (4- ((2- aminophenyls) carbamyl) benzyl) -1H- pyrroles [2,3-c] pyridine-2-carboxamide,
N- (4- ((2- aminophenyls) carbamyl) benzyl) -1H- pyrrolo-es [3,2-c] pyridine-2-carboxamide,
N- (4- ((2- aminophenyls) carbamyl) benzyl) -1H- pyrrolo-es [2,3-b] pyridine-2-carboxamide,
N- (4- ((2- aminophenyls) carbamoyl) benzyl) imidazo [1,5-a] pyridine -6- formamides,
N- (4- ((2- aminophenyls) carbamoyl) benzyl) -1H- pyrazolos [4,3-b] pyridine -6- formamides,
N- (4- ((2- aminophenyls) carbamoyl) benzyl) imidazoles [1,2-a] pyridine -7- formamides,
N- (2- aminophenyls) -4- ((4- (pyridin-3-yl) -1H-1,2,3- triazol-1-yls) methyl) benzamide,
N- (2- aminophenyls) -4- (2- (4- (pyridin-3-yl) -1H-1,2,3- triazol-1-yls) methyl) benzamide,
N- (2- aminophenyls) -4- ((4- (pyridin-4-yl) -1H-1,2,3- triazol-1-yls) methyl) benzamide,
N- (2- aminophenyls) -4- ((4- (6- chloropyridine -3- bases) -1H-1,2,3- triazol-1-yls) methyl) benzamide,
N- (2- aminophenyls) -4- ((4- (3- aminophenyls) -1H-1,2,3- triazol-1-yls) methyl) benzamide,
N- (2- aminophenyls) -4- ((4- (tolyl) -1H-1,2,3- triazol-1-yls) methyl) benzamide,
4- ((4- (4- acetylphenyls) -1H-1,2,3- triazol-1-yls) methyl)-N- (2- aminophenyls) benzamide,
N- (2- aminophenyls) -4- ((4- (thiophene -2- bases) -1H-1,2,3- triazol-1-yls) methyl) benzamide,
N- (4- ammonia-[1,1 '-biphenyl] -3- bases) -4- ((5- (pyridin-3-yl) -1H-1,2,3- triazol-1-yls) methyl) benzene first Acid amides,
N- (4- amino-[1,1 '-biphenyl] -3- bases) -4- ((5- (3- aminophenyls) -1H-1,2,3- triazol-1-yls) methyl) benzene Formamide,
N- (4- amino-[1,1 '-biphenyl] -3- bases) -4- ((5- (3- hydroxy phenyls) -1H-1,2,3- triazol-1-yls) methyl) benzene Formamide,
4- ((5- (4- acetylphenyls) -1H-1,2,3- triazol-1-yls) methyl)-N- (4- amino-[1,1 '-biphenyl] -3- bases) Benzamide,
N- (2- aminophenyls) -4- ((4- (4- (3- (pyridin-3-yl methyl) -1H-1,2,3- triazol-1-yls) methyl) benzoyls Amine,
N- (2- aminophenyls) -4- ((4- (4- ((pyridin-3-yl methyl) carbamyl) phenyl) -1H-1,2,3- triazole -1- Base) methyl) benzamide,
N- (2- aminophenyls) -8- (4- (pyridin-3-yl) -1H-1,2,3- triazol-1-yls) caprylamide,
N- (2- aminophenyls) -8- (4- (3- aminophenyls) -1H-1,2,3- triazol-1-yls) caprylamide,
N- (4- amino-[1,1 '-biphenyl] -3- bases) -8- (4- (pyridin-3-yl) -1H-1,2,3- triazol-1-yls) caprylamide,
N- (4- amino-[1,1 '-biphenyl] -3- bases) -8- (4- (3- aminophenyls) -1H-1,2,3- triazol-1-yls) caprylamide,
N- (4- amino-[1,1 '-biphenyl] -3- bases) -8- (4- (3- hydroxy phenyls) -1H-1,2,3- triazol-1-yls) caprylamide.
7. the preparation method of a kind of amide derivatives according to claim 1 and its pharmaceutically acceptable salt, it is special Levy and be：
When general structure isAnd X2For O or S, R are-NH-CH2- A ,-CH=CH-A orA For phenyl, amino substituted-phenyl, halogen substituted phenyl, pyridine radicals, the straight chained alkyl substituted pyridinyl of 1-6 carbon atom, 3-6 The branched alkyl substituted pyridinyl of carbon atom, halogen substituted pyridinyl, amino substituted pyridinyl, When, methods described comprises the following steps：
A1, cyano compound 1It is catalyzed through Raney Ni, hydrogen reducing obtains amino chemical combination in ammoniacal liquor methanol solution Thing 2aX1For N or CH, R2 are hydrogen, halogen, amino or t-butoxycarbonyl amino；
A2, p-aminobenzoic acid and CDI or TCDI react obtain intermediate 4 at room temperature
A3, compound 2a or compound 2cRoom temperature reaction obtains intermediate 5 in tetrahydrofuran with intermediate 4
Or, compound 2bIntermediate 5 is condensed to yield in dichloromethane with p-aminobenzoic acid；R3For H or C1；
A4, intermediate 5 in DMF with o-phenylenediamine room temperature reaction, obtain target compound through column chromatography
When general structure isAnd A is When, methods described includes following step Suddenly：
B1,4- amine methyl toluate and carboxylic acid obtain midbody compound 8 through EDC/DMAP catalyzing and condensings
B2, in tetrahydrofuran/methanol/water mixed solvent system, the midbody compound 8 through LiOH hydrolysis obtain Carboxylation Compound 9
B3, the carboxylic acid compound 9 react at ambient temperature in DMF with o-phenylenediamine, obtain final product target compound
When general structure isAnd n=1 or 2, A are pyridine radicals, halogen substituted pyridines Base, amino substituted pyridinyl, 1-6 carbon atom straight chain alkyl-substituted phenyl, acetyl group substituted-phenyl, thienyl,When Z is hydrogen atom or phenyl, methods described comprises the following steps：
C1, compound 11Compound 12 is reacted to obtain with trimethyl silicane ethyl-acetyleneIn the basic conditions The de- trimethyl silicon substrate of room temperature is obtained compound 13The R4 is chlorine or hydrogen；
C2, bromide 14Intermediate 15 is obtained through sodium azide substitution
Intermediate product 16 obtained and click reactions with compound 13 or other alkynyl compounds in C3, intermediate 15 there is
C4, the intermediate product 16 are hydrolyzed in the basic conditions, with o-phenylenediamine or the final acquisition of 4- phenyl o-phenylenediamine condensation Targeted and thing
When formula isAnd R ' is amino, hydroxyl or hydrogen atom, X3It is N Or CH, Z be hydrogen atom or phenyl when, methods described comprises the following steps：
D1, compound 19It is catalyzed in HBTU and triethylamine with o-phenylenediamine or 4- phenyl o-phenylenediamine Lower generation condensation reaction obtains intermediate 20
Target compound obtained and Click reactions in the aryl ethane of D2, intermediate 20 containing different substituents there is
8. a kind of amide derivatives according to claim 1 and its pharmaceutically acceptable salt are preparing niacinamide phosphoric acid Purposes in the double target spot inhibitor of phosphoribosynltransferase inhibitor, histon deacetylase (HDAC) inhibitor or HDAC/NAMPT.
9. a kind of amide derivatives according to claim 1 and its pharmaceutically acceptable salt are preparing antineoplastic In purposes.
10. purposes according to claim 9, it is characterised in that the tumour is liver cancer, lung cancer, intestinal cancer, oophoroma, preceding Row gland cancer, stomach cancer or lymph cancer.
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Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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WO2023185752A1 (en) * 2022-03-28 2023-10-05 上海科技大学 Triazolamide compound, preparation method therefor, and use thereof

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000256194A (en) * 1999-01-06 2000-09-19 Mitsui Chemicals Inc Intranuclear receptor agonist and agent for enhancing its effect
EP1437346A1 (en) * 1996-09-30 2004-07-14 Schering AG Benzamide derivatives useful as cell differentiation inducers
CN101633638A (en) * 2008-06-20 2010-01-27 江苏国华投资有限公司 Class I histone deacetylase inhibitor and application thereof
CN1997649B (en) * 2004-03-26 2010-08-11 梅特希尔基因公司 Inhibitors of histone deacetylase
CN102869261A (en) * 2010-03-01 2013-01-09 瑞科西有限公司 Compounds and therapeutic uses thereof
CN103420917A (en) * 2012-05-18 2013-12-04 国药一心制药有限公司 Benzamide compounds containing fused ring structure and applications thereof as antitumor medicaments

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1437346A1 (en) * 1996-09-30 2004-07-14 Schering AG Benzamide derivatives useful as cell differentiation inducers
JP2000256194A (en) * 1999-01-06 2000-09-19 Mitsui Chemicals Inc Intranuclear receptor agonist and agent for enhancing its effect
CN1997649B (en) * 2004-03-26 2010-08-11 梅特希尔基因公司 Inhibitors of histone deacetylase
CN101633638A (en) * 2008-06-20 2010-01-27 江苏国华投资有限公司 Class I histone deacetylase inhibitor and application thereof
CN102869261A (en) * 2010-03-01 2013-01-09 瑞科西有限公司 Compounds and therapeutic uses thereof
CN103420917A (en) * 2012-05-18 2013-12-04 国药一心制药有限公司 Benzamide compounds containing fused ring structure and applications thereof as antitumor medicaments

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
ACS: "RN：1347720-27-7", 《STN-REGISTRY数据库》 *
ACS: "RN：1829502-72-8", 《STN-REGISTRY数据库》 *
ACS: "RN：1829502-73-9", 《STN-REGISTRY数据库》 *
何世鹏: "基于HDAC多靶点抑制剂的设计、合成与抗肿瘤活性研究", 《福建中医药大学硕士学位论文》 *
尹子卉，等: "新型抗肿瘤组蛋白去乙酰化酶抑制剂西达本胺的合成", 《中国新药杂志》 *

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