WO2019015635A1 - Benzimidazole derivative, preparation method therefor and medical use thereof - Google Patents

Abstract

The present invention relates to a benzimidazole derivative, a preparation method therefor and the medical use thereof. In particular, the present invention relates to a benzimidazole derivative as shown in formula (I), a preparation method therefor and a pharmaceutically acceptable salt thereof and the use of same as a therapeutic agent, especially as a bromodomain protein inhibitor, wherein the definition of each substituent in formula (I) is the same as the definition in the description.

Description

苯并咪唑类衍生物及其制备方法及其在医药上的用途Benzimidazole derivatives, preparation method thereof and use thereof in medicine

本申请要求于2017年7月21日提交中国专利局、申请号为201710599222.3、发明名称为“苯并咪唑类衍生物及其制备方法及其在医药上的用途”的中国专利申请的优先权，其全部内容通过引用结合在本申请中。This application claims the priority of the Chinese Patent Application filed on July 21, 2017, the Chinese Patent Office, Application No. 201710599222.3, entitled "Benzimidazole Derivatives, Preparation Methods and Their Uses in Medicine", The entire contents of this application are incorporated herein by reference.

技术领域Technical field

本发明属于医药领域，具体涉及一种新的苯并咪唑类衍生物、其制备方法及含有该衍生物的药物组合物，以及它们作为治疗剂、特别是作为溴结构域蛋白抑制剂的用途。The invention belongs to the field of medicine, and in particular relates to a novel benzimidazole derivative, a preparation method thereof and a pharmaceutical composition containing the same, and their use as therapeutic agents, in particular as bromodomain protein inhibitors.

背景技术Background technique

近年来，肿瘤成为全球范围内导致人类死亡的主要原因之一。肿瘤普遍具有总体治愈率低且复发率高等特点，因此预防、治疗以及抑制肿瘤复发具有重要的科研价值，实现肿瘤的预防和治愈具有相当的紧迫性和挑战性。表观遗传调控的异常是导致肿瘤发生的重要因素之一。In recent years, cancer has become one of the leading causes of human death worldwide. Tumors generally have the characteristics of low overall cure rate and high recurrence rate. Therefore, prevention, treatment and inhibition of tumor recurrence have important scientific research value. It is quite urgent and challenging to achieve tumor prevention and cure. Abnormal epigenetic regulation is one of the important factors leading to tumorigenesis.

溴结构域(bromodomain)是一种能识别N-乙酰化赖氨酸残基的蛋白结构域。含溴结构域的蛋白质的BET家族包括四个成员(BRD2、BRD3、BRD4和BRDt)。BET家族的每个成员都使用两个溴结构域来识别，主要(但并非排外地)在组蛋白蛋白质的氨基末端尾巴(amino-terminal tail)发现N-乙酰化的赖氨酸残基。通过将转录因子募集至染色质内的具体基因组位置来调节基因表达。例如，组蛋白连接的BRD4将转录因子P-TEFb募集至启动子，导致表达涉及细胞周期进程的基因子集(Yang et al.,Mol.Cell.Biol.28:967-976(2008))。BRD2和BRD3还起生长促进基因的转录调节剂的作用。近来的研究已经证实以BET溴结构域为靶点来治疗多种癌症((Zuber et al.,Nature 478:524-528(2011)；Mertz et al.,Proc.Natl.Acad.Sci.108:16669-16674(2011)；Delmore et al.,Cell 146:1-14,(2011)；Dawson et al.,Nature 478:529-533(2011))、动脉粥样硬化、炎症(Huang et al.,Mol.Cell.Biol.29:1375-1387(2009))和HIV感染。The bromodomain is a protein domain that recognizes N-acetylated lysine residues. The BET family of bromodomain-containing proteins includes four members (BRD2, BRD3, BRD4, and BRDt). Each member of the BET family uses two bromodomains to recognize, primarily (but not exclusively) the discovery of N-acetylated lysine residues at the amino-terminal tail of histone proteins. Gene expression is regulated by recruiting transcription factors to specific genomic locations within the chromatin. For example, histone-linked BRD4 recruits the transcription factor P-TEFb to a promoter, resulting in the expression of a subset of genes involved in cell cycle progression (Yang et al., Mol. Cell. Biol. 28: 967-976 (2008)). BRD2 and BRD3 also function as transcriptional regulators of growth-promoting genes. Recent studies have demonstrated the targeting of BET bromodomains to treat a variety of cancers (Zuber et al., Nature 478: 524-528 (2011); Mertz et al., Proc. Natl. Acad. Sci. 108: 16669-16674 (2011); Delmore et al., Cell 146: 1-14, (2011); Dawson et al., Nature 478: 529-533 (2011)), atherosclerosis, inflammation (Huang et al. , Mol. Cell. Biol. 29: 1375-1387 (2009)) and HIV infection.

最新研究发现，BRD4蛋白介导的表观遗传异常与癌基因的过表达密切相关，并与癌细胞的生长增殖关系密切。BRD4是含溴结构域和额外终端域家族蛋白(Bromo and extra C-terminal domain,BET)蛋白家族的一员，由于在抗肿瘤方面的潜在价值，引起了各大制药公司和科研机构的极大关注。近期还发现BRD4在病毒基因的转录调控中也扮演了重要角色，并且与病毒瘤的发病机制存在一定联系。这些研究结果说明BRD4与多种肿瘤存在密切联系，尤其在一些至今 难以治愈或者尚无有效治疗手段的肿瘤中具有重要作用，其与肿瘤关系的研究为肿瘤治疗提供了新的策略。通过作用于BRD4蛋白溴结构域的小分子化合物，干扰溴结构域与乙酰化赖氨酸的特异性结合，影响肿瘤细胞内的转录调节和其它细胞过程，可以实现对肿瘤的靶向治疗。因此，BRD4蛋白是一个非常有前景的表观遗传新靶点，而作用于BRD4蛋白溴结构域的小分子抑制剂在肿瘤研究中也有着广阔的应用前景，而且有可能从中开发出新型抗肿瘤药物。The latest study found that BRD4 protein-mediated epigenetic abnormalities are closely related to overexpression of oncogenes and are closely related to the growth and proliferation of cancer cells. BRD4 is a member of the Bromo and extra C-terminal domain (BET) family of proteins, which has caused great potential for major pharmaceutical companies and research institutions due to its potential value in anti-tumor. attention. It has also recently been found that BRD4 also plays an important role in the transcriptional regulation of viral genes and is associated with the pathogenesis of viral tumors. These findings suggest that BRD4 is closely related to a variety of tumors, especially in some tumors that have been difficult to cure or have no effective treatment, and the study of its relationship with tumors provides a new strategy for cancer treatment. By targeting small molecule compounds of the BRD4 protein bromodomain, interfering with the specific binding of the bromodomain to acetylated lysine, affecting transcriptional regulation and other cellular processes in tumor cells, targeted therapy of tumors can be achieved. Therefore, BRD4 protein is a very promising new epigenetic target, and small molecule inhibitors acting on the bromodomain of BRD4 protein have broad application prospects in tumor research, and it is possible to develop a new type of anti-tumor. drug.

目前已经公开了一系列的溴结构域蛋白抑制剂专利，其中包括WO2011054846、WO2008092231、WO2012075383和WO2016139292等，其中WO2016139292公开了实施例1化合物。目前处于临床III期的药物为Apabetalone，处于临床II期的药物包括GSK-525762A、INCB-54329和BMS-986158等，同时有多个药物处于临床I期的药物。但这些对于抗肿瘤的研究是远远不够的，现有技术中公开的化合物以及试验药物在有效性、安全性或适用性等方面仍不能令人满意，仍有必要研究和开发新的溴结构域蛋白抑制剂，以满足人们日益增长的医疗和健康需要。A series of bromodomain protein inhibitor patents have been disclosed, including WO2011054846, WO2008092231, WO2012075383, and WO2016139292, among which WO2016139292 discloses the compound of Example 1. The drug currently in clinical stage III is Apabetalone, and the drugs in clinical phase II include GSK-525762A, INCB-54329 and BMS-986158, and there are several drugs in clinical phase I. However, these studies on anti-tumor are far from enough. The compounds disclosed in the prior art and the test drugs are still unsatisfactory in terms of effectiveness, safety or applicability. It is still necessary to study and develop new bromo structures. Domain protein inhibitors to meet the growing medical and health needs of people.

发明内容Summary of the invention

本发明人通过实验研究意外地发现，下式(I)的化合物可以有效用作溴结构域蛋白抑制剂，用于治疗或预防与溴结构域蛋白相关的疾病。The present inventors have unexpectedly discovered through experimental research that the compound of the following formula (I) can be effectively used as a bromodomain protein inhibitor for the treatment or prevention of a disease associated with a bromodomain protein.

因此，在第一个方面，本发明提供了式(I)所示的苯并咪唑类衍生物：Accordingly, in a first aspect, the present invention provides a benzimidazole derivative of the formula (I):

包括其立体异构体、互变异构体或其可药用的盐，Including stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

其中：among them:

X ¹选自-CH＝或-N＝； X ^{1 is} selected from -CH= or -N=;

X ²选自-CH＝或-N＝； X ^{2 is} selected from -CH= or -N=;

且X ¹和X ²不同时为-N＝； And X ¹ and X ^{2 are} not -N=;

R ¹选自烷基；优选为甲基； R ^{1 is} selected from an alkyl group; preferably a methyl group;

R ²选自氢原子或烷基；优选为甲基； R ^{2 is} selected from a hydrogen atom or an alkyl group; preferably a methyl group;

R ³选自烷基，其中所述的烷基进一步被一个或多个烷氧基所取代； R ^{3 is} selected from alkyl, wherein said alkyl group is further substituted with one or more alkoxy groups;

R ⁴选自卤素； R ^{4 is} selected from halogen;

R ⁵选自氢原子、烷基、氰基、卤素、烷氧基、环烷基、杂环基、芳基、杂芳基、-OR ¹⁰、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹，其中所述的烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹的取代基所取代； R ^{5 is} selected from the group consisting of a hydrogen atom, an alkyl group, a cyano group, a halogen, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, -OR ¹⁰ , -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , —C(O)R ¹⁰ , —C(O)OR ¹⁰ or —NR ⁸ C(O)R ⁹ , wherein said alkyl group, cycloalkyl group, heterocyclic group, aryl group or heteroaryl group Further optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -NR ⁸ R ^{9, -C (O) NR 8} R 9, -C (O) R 10, -C (O) oR 10 , or -NR ⁸ C (O) R ⁹ is substituted with a substituent;

R ⁶选自氢原子、烷基、氰基、烷氧基、环烷基、杂环基、芳基、杂芳基、-OR ¹⁰、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹，其中所述的烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹的取代基所取代； R ^{6 is} selected from the group consisting of a hydrogen atom, an alkyl group, a cyano group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, -OR ¹⁰ , -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , —C(O)R ¹⁰ , —C(O)OR ¹⁰ or —NR ⁸ C(O)R ⁹ , wherein said alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl is Further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -NR ⁸ R ⁹ , Substituted by a substituent of -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -C(O)OR ¹⁰ or -NR ⁸ C(O)R ⁹ ;

R ⁷选自氢原子或烷基；优选为氢原子； R ^{7 is} selected from a hydrogen atom or an alkyl group; preferably a hydrogen atom;

R ⁸、R ⁹和R ¹⁰各自独立地选自氢原子、烷基、环烷基、杂环基、芳基或杂芳基，其中所述烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ¹¹R ¹²、-C(O)NR ¹¹R ¹²、-C(O)R ¹³、-C(O)OR ¹³或-NR ¹¹C(O)R ¹²的取代基所取代； R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group Or a heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, - Substituted by a substituent of NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

或者，R ⁸和R ⁹与相连接的N原子一起形成一个4～8元杂环基，其中4～8元杂环内含有一个或多个N、O、S(O) _p原子，并且4～8元杂环上任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ¹¹R ¹²、-C(O)NR ¹¹R ¹²、-C(O)R ¹³、-C(O)OR ¹³或-NR ¹¹C(O)R ¹²的取代基所取代； Alternatively, R ⁸ and R ⁹ together with the N atom to which they are attached form a 4 to 8 membered heterocyclic group, wherein the 4 to 8 membered heterocyclic ring contains one or more N, O, S(O) _p atoms, and 4 Optionally, one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0 Substituting a substituent of -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

R ¹¹、R ¹²和R ¹³各自独立地选自氢原子、烷基、环烷基、杂环基、芳基或杂芳基，其中所述烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、羧基或羧酸酯基的取代基所取代；且 R ¹¹ , R ¹² and R ¹³ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group Or a heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxy or carboxylic acid Substituted by a substituent of an ester group;

p为0、1或2。p is 0, 1, or 2.

在本发明的一些优选方案中，所述式(I)化合物具有式(II)所示的结构，In some preferred embodiments of the invention, the compound of formula (I) has the structure shown in formula (II),

其中：among them:

R ^a和R ^b各自独立地选自氢原子或烷氧基，且R ^a和R ^b不同时为氢原子；所述的烷氧基优选为甲氧基； R ^a and R ^b are each independently selected from a hydrogen atom or an alkoxy group, and R ^a and R ^{b are} not simultaneously a hydrogen atom; the alkoxy group is preferably a methoxy group;

m为1、2、3或4；优选为1或2；m is 1, 2, 3 or 4; preferably 1 or 2;

n为1、2、3或4；优选为1或2；且n is 1, 2, 3 or 4; preferably 1 or 2;

R ¹、R ²、R ⁴～R ⁷的定义如通式(I)中所述。 R ¹ , R ² , and R ⁴ to R ⁷ are as defined in the formula (I).

在本发明的一些优选方案中，式(II)所示化合物具有式(III)所示结构，In some preferred embodiments of the present invention, the compound of the formula (II) has a structure represented by the formula (III).

其中：among them:

R ^a选为氢原子； R ^{a is} selected as a hydrogen atom;

R ^b选自烷氧基，优选为甲氧基； R ^{b is} selected from alkoxy groups, preferably methoxy groups;

m为1、2、3或4；优选为1或2；m is 1, 2, 3 or 4; preferably 1 or 2;

n为1、2、3或4；优选为1或2；且n is 1, 2, 3 or 4; preferably 1 or 2;

R ¹、R ²、R ⁴～R ⁷的定义如通式(I)中所述。 R ¹ , R ² , and R ⁴ to R ⁷ are as defined in the formula (I).

在本发明的一些优选方案中，式(II)所示化合物具有式(IV)所示结构，In some preferred embodiments of the present invention, the compound of the formula (II) has a structure represented by the formula (IV).

其中：among them:

R ^a为氢原子； R ^a is a hydrogen atom;

R ^b选自烷氧基，优选为甲氧基； R ^{b is} selected from alkoxy groups, preferably methoxy groups;

m为1、2、3或4；优选为1或2；m is 1, 2, 3 or 4; preferably 1 or 2;

n为1、2、3或4；优选为1或2；且n is 1, 2, 3 or 4; preferably 1 or 2;

R ¹、R ²、R ⁴～R ⁷的定义如通式(I)中所述。 R ¹ , R ² , and R ⁴ to R ⁷ are as defined in the formula (I).

在本发明的一些优选方案中，对式(I)、(II)、(III)或(IV)所示的化合物而言，R ⁴选自氟、氯或溴；优选为氟。 In some preferred embodiments of the invention, for the compound of formula (I), (II), (III) or (IV), R ^{4 is} selected from fluoro, chloro or bromo; preferably fluoro.

在本发明的一些优选方案中，对式(I)、(II)、(III)或(IV)所示的化合物而言，R ⁶选自杂环基，优选为吗啉基。 In some preferred embodiments of the invention, for the compound of formula (I), (II), (III) or (IV), R ^{6 is} selected from heterocyclyl, preferably morpholinyl.

本发明的典型化合物包括但不限于：Typical compounds of the invention include, but are not limited to:

，包括其立体异构体、互变异构体或其可药用的盐。Including stereoisomers, tautomers or pharmaceutically acceptable salts thereof.

进一步，本发明提供了式(I)化合物的制备方法，该方法包括：Further, the present invention provides a process for the preparation of a compound of formula (I), which process comprises:

使式(IA)化合物与R ⁶H反应，得到式(I)化合物； Reaction of a compound of formula (IA) with R ⁶ H to provide a compound of formula (I);

其中：among them:

X选自卤素，优选为氯或溴；X is selected from halogen, preferably chlorine or bromine;

R ⁶选自杂环基；且 R ^{6 is} selected from heterocyclic groups;

R ¹～R ⁵、R ⁷、X ¹和X ²的定义如通式(I)中所述。 R ¹ to R ⁵ , R ⁷ , X ¹ and X ² are as defined in the formula (I).

本发明还提供了式(IA)所示的化合物，包括其立体异构体、互变异构体或其可药用的盐，The present invention also provides a compound represented by the formula (IA), including stereoisomers, tautomers thereof or pharmaceutically acceptable salts thereof,

其中：among them:

X选自卤素，优选为氯或溴；且X is selected from halogen, preferably chlorine or bromine;

R ¹～R ⁵、R ⁷、X ¹和X ²的定义如通式(I)中所述。 R ¹ to R ⁵ , R ⁷ , X ¹ and X ² are as defined in the formula (I).

式(IA)的典型化合物包括但不限于：Typical compounds of formula (IA) include, but are not limited to:

包括其立体异构体、互变异构体或其可药用的盐。Included are stereoisomers, tautomers or pharmaceutically acceptable salts thereof.

进一步，本发明提供了式(IA)化合物的制备方法，该方法包括：Further, the present invention provides a process for the preparation of a compound of formula (IA), which process comprises:

使式(Ib)化合物与式(Ic)化合物反应，得到式(IA)化合物；Reaction of a compound of formula (Ib) with a compound of formula (Ic) to provide a compound of formula (IA);

其中：among them:

X选自卤素，优选为氯或溴；且X is selected from halogen, preferably chlorine or bromine;

R ¹～R ⁵、R ⁷、X ¹和X ²的定义如通式(I)中所述。 R ¹ to R ⁵ , R ⁷ , X ¹ and X ² are as defined in the formula (I).

更近一步，本发明提供了一种药物组合物，所述的药物组合物含有有效剂量的式(I)、(II)、(III)或(IV)所述的化合物(包括立体异构体、互变异构体或其可药用的盐)，以及可药用的载体、赋形剂或它们的组合。Further, the present invention provides a pharmaceutical composition comprising an effective amount of a compound of formula (I), (II), (III) or (IV) (including stereoisomers) And tautomers or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers, excipients or combinations thereof.

本发明还提供了一种抑制溴结构域蛋白的方法，包括将所述的蛋白与式(I)、(II)、(III)或(IV)所述的化合物或其药物组合物相接触，其中所述的溴结构域蛋白优选为BRD2、BRD3和BRD4，更优选为BRD4。The present invention also provides a method of inhibiting a bromodomain protein comprising contacting the protein with a compound of formula (I), (II), (III) or (IV) or a pharmaceutical composition thereof, The bromodomain proteins described therein are preferably BRD2, BRD3 and BRD4, more preferably BRD4.

本发明进一步提供了式(I)、(II)、(III)或(IV)所述的化合物(包括其立体异构体、互变异构体或其可药用的盐)或其药物组合物在制备用作溴结构域蛋白抑制剂的药物中的用途，其中所述的溴结构域蛋白优选为BRD2、BRD3和BRD4，更优选为BRD4。The invention further provides a compound of formula (I), (II), (III) or (IV), including stereoisomers, tautomers or pharmaceutically acceptable salts thereof, or a pharmaceutical combination thereof Use of a substance for the preparation of a medicament for use as a bromodomain protein inhibitor, wherein said bromodomain protein is preferably BRD2, BRD3 and BRD4, more preferably BRD4.

本发明进一步提供了式(I)、(II)、(III)或(IV)所述的化合物(包括其立体异构体、互变异构体或其可药用的盐)或其药物组合物在制备用于治疗或预防与溴结构域蛋白相关的疾病的药物中的用途，其中所述的疾病优选为癌症或炎症，其中所述的炎症优选为类风湿性关节炎、克隆恩病、湿疹、巨细胞性动脉炎、肝炎、炎性肠病、骨关节炎、胰腺炎、肺炎、银屑病、银屑病性关节炎、***性红斑狼疮、肾小球性肾炎、狼疮性肾炎、膜性肾小球肾炎或心肌炎；所述的炎症更优选为类风湿性关节炎；其中所述的癌症优选为小细胞肺癌、非小细胞肺癌、乳腺癌、结直肠癌、***癌、黑色素瘤、胰腺癌、神经胶质瘤、脑瘤、***、卵巢癌、胰腺癌、***癌、肾细胞癌、胃癌、膀胱癌、肝癌、睾丸核蛋白中线癌、多发性骨髓瘤、急性髓性白血病、急性淋巴细胞性白血病、慢性淋巴细胞性白血病、慢性髓细胞性白血病或慢性骨髓性白血病。The invention further provides a compound of formula (I), (II), (III) or (IV), including stereoisomers, tautomers or pharmaceutically acceptable salts thereof, or a pharmaceutical combination thereof Use of a medicament for the treatment or prevention of a disease associated with a bromodomain protein, wherein the disease is preferably cancer or inflammation, wherein the inflammation is preferably rheumatoid arthritis, cloning enemies, Eczema, giant cell arteritis, hepatitis, inflammatory bowel disease, osteoarthritis, pancreatitis, pneumonia, psoriasis, psoriatic arthritis, systemic lupus erythematosus, glomerulonephritis, lupus nephritis, Membranous glomerulonephritis or myocarditis; the inflammation is more preferably rheumatoid arthritis; wherein the cancer is preferably small cell lung cancer, non-small cell lung cancer, breast cancer, colorectal cancer, prostate cancer, melanoma , pancreatic cancer, glioma, brain tumor, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, stomach cancer, bladder cancer, liver cancer, testicular nucleoprotein, cancer, multiple myeloma, acute myeloid leukemia Acute lymphocytic Leukemia, chronic lymphocytic leukemia, chronic myelogenous leukemia, or chronic myelogenous leukemia.

本发明还提供了式(I)、(II)、(III)或(IV)所述的化合物(包括其立体异构体、互变异构体或其可药用的盐)或其药物组合物在制备用于治疗或预防糖尿病性肾病、高血压性肾病、HIV-相关的肾病、多囊性肾病、肥胖、血脂异常、高胆固醇血症、阿尔茨海默病、代谢综合征、脂肪肝、II型糖尿病、胰岛素抵抗、糖尿病性视网膜病或糖尿病性神经病的药物中的用途。The present invention also provides a compound of the formula (I), (II), (III) or (IV), including stereoisomers, tautomers or pharmaceutically acceptable salts thereof, or a pharmaceutical combination thereof Preparation for the treatment or prevention of diabetic nephropathy, hypertensive nephropathy, HIV-related nephropathy, polycystic kidney disease, obesity, dyslipidemia, hypercholesterolemia, Alzheimer's disease, metabolic syndrome, fatty liver Use in drugs for type 2 diabetes, insulin resistance, diabetic retinopathy or diabetic neuropathy.

发明的详细说明Detailed description of the invention

除非有相反陈述，否则本发明在说明书和权利要求书中所使用的部分术语 定义如下：Unless otherwise stated, some of the terms used in the specification and claims of the invention are defined as follows:

“烷基”当作一基团或一基团的一部分时是指包括直链或者带有支链的C ₁-C ₂₀脂肪烃基团。优选为C ₁-C ₁₀烷基，更优选为C ₁-C ₆烷基，特别优选为C ₁-C ₄烷基。烷基基团的实施例包括但不限于甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基等。烷基可以是取代或未取代的。 When "alkyl" as a group or part of a group is meant to include a straight-chain or branched with C ₁ -C ₂₀ aliphatic hydrocarbon group. It is preferably a C ₁ -C ₁₀ alkyl group, more preferably a C ₁ -C ₆ alkyl group, and particularly preferably a C ₁ -C ₄ alkyl group. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, sec-butyl, n-pentyl, 1, 1-di Methylpropyl, 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-dimethyl Butyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl Wait. The alkyl group can be substituted or unsubstituted.

“环烷基”是指饱和或部分饱和的单环、稠环、桥环或螺环的碳环。优选为C ₃-C ₁₂环烷基，更优选为C ₃-C ₈环烷基，最优选为C ₃-C ₆环烷基。单环环烷基的实施例包括但不限于环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基、环庚三烯基、环辛基等，优选环丙基、环己烯基。 "Cycloalkyl" means a saturated or partially saturated monocyclic, fused, bridged or spiro carbon ring. It is preferably a C ₃ -C ₁₂ cycloalkyl group, more preferably a C ₃ -C ₈ cycloalkyl group, and most preferably a C ₃ -C ₆ cycloalkyl group. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene The alkenyl group, the cyclooctyl group and the like are preferably a cyclopropyl group or a cyclohexenyl group.

“螺环烷基”指5至18元的、含有两个或两个以上环状结构的且单环之间彼此共用一个碳原子(称螺原子)的多环基团，环内可含有1个或多个双键，但没有一个环具有完全共轭的π电子的芳香***。优选为6至14元，更优选为7至10元。根据环与环之间共用螺原子的数目将螺环烷基分为单螺、双螺或多螺环烷基，优选为单螺和双螺环烷基，优选为4元/5元、4元/6元、5元/5元或5元/6元。“螺环烷基”的非限制性实施例包括但不限于：螺[4.5]癸基、螺[4.4]壬基、螺[3.5]壬基、螺[2.4]庚基。"Spirocycloalkyl" means a 5- to 18-membered polycyclic group having two or more cyclic structures and sharing a carbon atom (called a spiro atom) with each other, and the ring may contain 1 One or more double bonds, but none of the rings have a fully conjugated π-electron aromatic system. It is preferably 6 to 14 members, more preferably 7 to 10 members. The spirocycloalkyl group is classified into a monospiro, a spiro- or a spirocycloalkyl group, preferably a mono- and bi-spirocycloalkyl group, preferably 4 yuan/5 yuan, 4, depending on the number of common spiro atoms between the rings. Yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6 yuan. Non-limiting examples of "spirocycloalkyl" include, but are not limited to, spiro[4.5]decyl, spiro[4.4]decyl, spiro[3.5]decyl, spiro[2.4]heptyl.

“稠环烷基”指5至18元的、含有两个或两个以上环状结构的彼此共用一对碳原子的全碳多环基团，其中一个或多个环可以含有一个或多个双键，但没有一个环具有完全共轭的π电子的芳香***，优选为6至12元，更优选为7至10元。根据组成环的数目可以分为双环、三环、吡啶酮或多环稠环烷基，优选为双环或三环，更优选为5元/5元或5元/6元双环烷基。“稠环烷基”的非限制性实施例包括但不限于：二环[3.1.0]己基、二环[3.2.0]庚-1-烯基、二环[3.2.0]庚基、十氢化萘基或十四氢菲基。"Fused cycloalkyl" refers to a 5 to 18 membered all carbon polycyclic group having two or more cyclic structures that share a pair of carbon atoms with each other, wherein one or more of the rings may contain one or more A double bond, but none of the rings have a fully conjugated π-electron aromatic system, preferably 6 to 12 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic fused ring alkyl group, preferably a bicyclic ring or a tricyclic ring, more preferably a 5-membered/5-membered or 5-membered/6-membered bicycloalkyl group. Non-limiting examples of "fused cycloalkyl" include, but are not limited to, bicyclo[3.1.0]hexyl, bicyclo[3.2.0]hept-1-enyl, bicyclo[3.2.0]heptyl, Decalinyl or tetradecafluorophenanyl.

“桥环烷基”指5至18元的、含有两个或两个以上环状结构的彼此共用两个不直接相连接碳原子的全碳多环基团，其中一个或多个环可以含有一个或多个双键，但没有一个环具有完全共轭的π电子的芳香***。优选为6至14元，更优选为7至10元。根据组成环的数目可以分为双环、三环、吡啶酮或多环桥环烷基，优选为双环、三环或吡啶酮，更有选为双环或三环。“桥环烷基”的非限制性实施例包括但不限于：(1s,4s)-二环[2.2.1]庚基、二环[3.2.1]辛基、(1s,5s)-二环 [3.3.1]壬基、二环[2.2.2]辛基、(1r,5r)-二环[3.3.2]癸基。"Bridge cycloalkyl" refers to a 5- to 18-membered, all-carbon polycyclic group containing two or more cyclic structures that share two non-directly bonded carbon atoms, wherein one or more rings may contain One or more double bonds, but none of the rings have a fully conjugated π-electron aromatic system. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic bridged cycloalkyl group, preferably a bicyclic ring, a tricyclic ring or a pyridone, and more preferably a bicyclic ring or a tricyclic ring. Non-limiting examples of "bridged cycloalkyl" include, but are not limited to: (1s, 4s)-bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl, (1s,5s)-di Ring [3.3.1] fluorenyl, bicyclo [2.2.2] octyl, (1r, 5r)-bicyclo[3.3.2] fluorenyl.

所述环烷基环可以稠合于芳基、杂芳基或杂环基环上，其中与母体结构连接在一起的环为环烷基，非限制性实施例包括茚满基、四氢萘基、苯并环庚烷基等。环烷基可以是任选取代的或未取代的。The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocyclyl ring, wherein the ring to which the parent structure is attached is a cycloalkyl group, non-limiting examples include indanyl, tetrahydronaphthalene Base, benzocycloheptyl and the like. The cycloalkyl group can be optionally substituted or unsubstituted.

“杂环基”、“杂环”或“杂环的”在本申请中可交换使用，都是指非芳香性杂环基，其中一个或多个成环的原子是杂原子，如氧、氮、硫原子等，包括单环、稠环、桥环和螺环。优选具有5至7元单环或7至10元双-或三环，其可以包含1，2或3个选自氮、氧和/或硫中的原子。“杂环基”的实例包括但不限于吗啉基，氧杂环丁烷基，硫代吗啉基，四氢吡喃基，1,1-二氧代-硫代吗啉基，哌啶基，2-氧代-哌啶基，吡咯烷基，2-氧代-吡咯烷基，哌嗪-2-酮，8-氧杂-3-氮杂-双环[3.2.1]辛基和哌嗪基。杂环基可以是取代或未取代的。"Heterocyclyl", "heterocyclic" or "heterocyclic" are used interchangeably herein to refer to a non-aromatic heterocyclic group wherein one or more of the ring-forming atoms are heteroatoms such as oxygen, Nitrogen, sulfur atoms, etc., including monocyclic, fused, bridged, and spiro rings. It preferably has a 5- to 7-membered monocyclic ring or a 7- to 10-membered double- or tricyclic ring which may contain 1, 2 or 3 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heterocyclyl" include, but are not limited to, morpholinyl, oxetane, thiomorpholinyl, tetrahydropyranyl, 1,1-dioxo-thiomorpholinyl, piperidine , 2-oxo-piperidinyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, piperazin-2-one, 8-oxa-3-aza-bicyclo[3.2.1]octyl and Piperazinyl. The heterocyclic group may be substituted or unsubstituted.

“螺杂环基”指5至18元的、含义两个或两个以上环状结构的且单环之间彼此共用一个原子的多环基团，其环内含有1个或多个双键，但没有一个环具有完全共轭的π电子的芳香***，其中一个或多个环原子选自氮、氧或S(O) _p(其中p选自0、1或2)的杂原子，其余环原子为碳。优选为6至14元，更优选为7至10元。根据环与环之间共用螺原子的数目将螺环烷基分为单螺杂环基、双螺杂环基或多螺杂环基，优选为单螺杂环基和双螺杂环基。更优选为4元/4元、4元/5元、4元/6元、5元/5元或5元/6元单螺杂环基。“螺杂环基”的非限制性实施例包括但不限于：1,7-二氧杂螺[4.5]癸基、2-氧杂-7-氮杂螺[4.4]壬基、7-氧杂螺[3.5]壬基和5-氧杂螺[2.4]庚基。 "spiroheterocyclyl" means a 5- to 18-membered polycyclic group having the meaning of two or more cyclic structures and sharing one atom with each other, having one or more double bonds in the ring. , but none of the rings have a fully conjugated π-electron aromatic system in which one or more ring atoms are selected from nitrogen, oxygen or S(O) _p (where p is selected from 0, 1 or 2) heteroatoms, the remainder The ring atom is carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. The spirocycloalkyl group is classified into a monospiroheterocyclic group, a dispiroheterocyclic group or a polyspirocyclic group according to the number of common spiro atoms between the ring and the ring, and is preferably a monospiroheterocyclic group and a dispiroheterocyclic group. More preferably, it is 4 yuan / 4 yuan, 4 yuan / 5 yuan, 4 yuan / 6 yuan, 5 yuan / 5 yuan or 5 yuan / 6-membered monospiroheterocyclic group. Non-limiting examples of "spiroheterocyclyl" include, but are not limited to, 1,7-dioxaspiro[4.5]fluorenyl, 2-oxa-7-azaspiro[4.4]decyl, 7-oxo Heterospiro[3.5]decyl and 5-oxaspiro[2.4]heptyl.

“稠杂环基”指含有两个或两个以上环状结构彼此共用一对原子的全碳多环基团，其中一个或多个环可以含有一个或多个双键，但没有一个环具有完全共轭的π电子的芳香***，其中一个或多个环原子为选自氮、氧或S(O) _p(其中p选自0、1或2)的杂原子，其余环原子为碳。优选为6至14元，更优选为7至10元。根据组成环的数目可以分为双环、三环、吡啶酮或多环稠杂环基，优选为双环或三环，更优选为5元/5元或5元/6元双环稠杂环基。“稠杂环基”的非限制性实施例包括但不限于：八氢吡咯并[3,4-c]吡咯基、八氢-1H-异吲哚基，3-氮杂二环[3.1.0]己基，八氢苯并[b][1,4]二噁英(dioxine)。 "Fused heterocyclic group" refers to an all-carbon polycyclic group containing two or more cyclic structures that share a pair of atoms with each other, wherein one or more of the rings may contain one or more double bonds, but none of the rings have A fully conjugated π-electron aromatic system in which one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O) _p (where p is selected from 0, 1 or 2) and the remaining ring atoms are carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic fused heterocyclic group, preferably a bicyclic ring or a tricyclic ring, and more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group. Non-limiting examples of "fused heterocyclic groups" include, but are not limited to, octahydropyrrolo[3,4-c]pyrrolyl, octahydro-1H-isoindenyl, 3-azabicyclo[3.1. 0] hexyl, octahydrobenzo[b][1,4]dioxine.

“桥杂环基”指5至18元、优选5至14元，含有两个或两个以上环状结构且彼此共用两个不直接相连接的原子的多环基团，其中一个或多个环可以含有一个或多个双键，但没有一个环具有完全共轭的π电子的芳香***，其中一个或多个环原子选自氮、氧或S(O) _p(其中p选自0、1或2)的杂原子，其余环原子为碳。优选为6至14元，更优选为7至10元。根据组成环的数目可以分为双环、 三环、吡啶酮或多环桥杂环基，优选为双环、三环或吡啶酮，更有选为双环或三环。“稠杂环基”的非限制性实施例包括但不限于：2-氮杂二环[2.2.1]庚基，2-氮杂二环[2.2.2]辛基和2-氮杂二环[3.3.2]癸基。所述杂环基环可以稠合于芳基、杂芳基或环烷基环上，其中与母体结构连接在一起的环为杂环基。杂环基可以是任选取代的或未取代的。 "Bridge heterocyclyl" refers to a polycyclic group of 5 to 18 members, preferably 5 to 14 members, containing two or more cyclic structures and sharing two atoms which are not directly bonded to each other, one or more of which A ring may contain one or more double bonds, but none of the rings have a fully conjugated π-electron aromatic system in which one or more ring atoms are selected from nitrogen, oxygen or S(O) _p (where p is selected from 0, 1 or 2) of a hetero atom, the remaining ring atoms being carbon. It is preferably 6 to 14 members, more preferably 7 to 10 members. Depending on the number of constituent rings, it may be classified into a bicyclic ring, a tricyclic ring, a pyridone or a polycyclic bridged heterocyclic group, preferably a bicyclic ring, a tricyclic ring or a pyridone, and more preferably a bicyclic ring or a tricyclic ring. Non-limiting examples of "fused heterocyclic groups" include, but are not limited to, 2-azabicyclo[2.2.1]heptyl, 2-azabicyclo[2.2.2]octyl and 2-aza-di Ring [3.3.2] sulfhydryl. The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring wherein the ring to which the parent structure is attached is a heterocyclic group. The heterocyclic group may be optionally substituted or unsubstituted.

“芳基”是指含有一个或者两个环的碳环芳香***，其中所述环可以以稠合的方式连接在一起。术语“芳基”包括比如苯基、萘基、四氢萘基的芳香基团。优选芳基为C ₆-C ₁₀芳基，更优选芳基为苯基和萘基，最优选为苯基。芳基可以是取代或未取代的。所述“芳基”可与杂芳基、杂环基或环烷基稠合，其中与母体结构连接在一起的为芳基环，非限制性实施例包括但不限于： "Aryl" means a carbocyclic aromatic system containing one or two rings wherein the rings may be joined together in a fused manner. The term "aryl" includes aryl groups such as phenyl, naphthyl, tetrahydronaphthyl. Preferably, the aryl group is a C ₆ -C ₁₀ aryl group, more preferably the aryl group is a phenyl group and a naphthyl group, and most preferably a phenyl group. The aryl group can be substituted or unsubstituted. The "aryl" may be fused to a heteroaryl, heterocyclyl or cycloalkyl group, wherein the parent structure is attached to an aryl ring, non-limiting examples include, but are not limited to:

“杂芳基”是指芳香族5至6元单环或9至10元双环，其可以包含1至4个选自氮、氧和/或硫中的原子。“杂芳基”的实施例包括但不限于呋喃基，吡啶基，2-氧代-1，2-二氢吡啶基，哒嗪基，嘧啶基，吡嗪基，噻吩基，异噁唑基，噁唑基，噁二唑基，咪唑基，吡咯基，吡唑基，***基，四氮唑基，噻唑基，异噻唑基，1，2，3-噻二唑基，苯并间二氧杂环戊烯基，苯并咪唑基，吲哚基，异吲哚基，1，3-二氧代-异吲哚基，喹啉基，吲唑基，苯并异噻唑基，苯并噁唑基和苯并异噁唑基。杂芳基可以是取代或未取代的。所述杂芳基环可以稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环，非限制性实施例包括但不限于："Heteroaryl" means an aromatic 5 to 6 membered monocyclic or 9 to 10 membered bicyclic ring which may contain from 1 to 4 atoms selected from nitrogen, oxygen and/or sulfur. Examples of "heteroaryl" include, but are not limited to, furyl, pyridyl, 2-oxo-1,2-dihydropyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, thienyl, isoxazolyl , oxazolyl, oxadiazolyl, imidazolyl, pyrrolyl, pyrazolyl, triazolyl, tetrazolyl, thiazolyl, isothiazolyl, 1,2,3-thiadiazolyl, benzo Dioxolyl, benzimidazolyl, fluorenyl, isodecyl, 1,3-dioxo-isoindenyl, quinolyl, oxazolyl, benzisothiazolyl, benzene And oxazolyl and benzoisoxazolyl. Heteroaryl groups can be substituted or unsubstituted. The heteroaryl ring can be fused to an aryl, heterocyclic or cycloalkyl ring wherein the ring to which the parent structure is attached is a heteroaryl ring, non-limiting examples include, but are not limited to:

“烷氧基”是指(烷基-O-)的基团。其中，烷基见本文有关定义。C ₁-C ₆的烷氧基为优先选择，尤其优选C ₁-C ₄烷氧基。其实例包括，但不限于：甲氧基、乙氧基、正丙氧基、异丙氧基、正丁氧基、异丁氧基、叔丁氧基等。 "Alkoxy" means a group of (alkyl-O-). Among them, the alkyl group is defined in the relevant definition herein. The C ₁ -C ₆ alkoxy group is preferred, and a C ₁ -C ₄ alkoxy group is particularly preferred. Examples thereof include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy and the like.

“羟基”指-OH基团。"Hydroxy" refers to an -OH group.

“卤素”是指氟、氯、溴和碘。"Halogen" means fluoro, chloro, bromo and iodo.

“氨基”指-NH ₂。 "Amino" means -NH ₂ .

“氰基”指-CN。"Cyano" means -CN.

“硝基”指-NO ₂。 "Nitro" means -NO ₂ .

“苄基”指-CH ₂-苯基。 "Benzyl" refers to -CH ₂ - phenyl.

“羧基”指-C(O)OH。"Carboxy" refers to -C(O)OH.

“羧酸酯基”指-C(O)O(烷基)或(环烷基)，其中烷基、环烷基的定义如上所述。"Carboxylic acid ester group" means -C(O)O(alkyl) or (cycloalkyl) wherein alkyl, cycloalkyl are as defined above.

“Bn”指苄基。"Bn" means benzyl.

“DMSO”指二甲基亚砜。"DMSO" refers to dimethyl sulfoxide.

“取代的”指基团中的一个或多个氢原子，优选为最多5个，更优选为1～3个氢原子彼此独立地被相应数目的取代基替换。不言而喻，取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和键(如烯键)的碳原子结合时可能是不稳定的。"Substituted" refers to one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3, hydrogen atoms are replaced by a corresponding number of substituents independently of one another. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated bond such as an ethylenic bond.

本说明书所述的“取代”或“取代的”，如无特别指出，均是指基团可被一个或多个选自以下的基团取代：烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氨基、卤代烷基、羟烷基、羧基、羧酸酯基、＝O、-OR ¹⁰、-SR ¹⁰、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹。 As used herein, "substituted" or "substituted", unless otherwise indicated, means that the group may be substituted by one or more groups selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy. , alkylthio, alkylamino, halogen, fluorenyl, hydroxy, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Base, heterocycloalkylthio, amino, haloalkyl, hydroxyalkyl, carboxyl, carboxylate, =0, -OR ¹⁰ , -SR ¹⁰ , -C(O)NR ⁸ R ⁹ , -C(O) R ¹⁰ , -C(O)OR ¹⁰ or -NR ⁸ C(O)R ⁹ .

“可药用的盐”是指上述化合物能保持原有生物活性并且适合于医药用途的某些盐类。式(I)化合物的可药用的盐可以为金属盐、与合适的酸形成的胺盐，金属盐优选碱金属、碱土金属盐，合适的酸包括无机酸和有机酸。"Pharmaceutically acceptable salt" refers to certain salts of the above compounds which retain their original biological activity and are suitable for pharmaceutical use. The pharmaceutically acceptable salt of the compound of the formula (I) may be a metal salt, an amine salt formed with a suitable acid, a metal salt preferably an alkali metal or an alkaline earth metal salt, and suitable acids include inorganic acids and organic acids.

“药物组合物”表示含有一种或多种本文所述化合物(包括其生理学上可药用的盐或立体异构体、互变异构体或前体药物等形式)与任选的其他药物活性成分的混合物，其可以包含其他任选组分例如可药用的载体和/或赋形剂。药物组合物的目的是促进对生物体的给药，利于活性成分的吸收进而发挥生物活性。"Pharmaceutical composition" means a compound comprising one or more of the compounds described herein, including physiologically pharmaceutically acceptable salts or stereoisomers, tautomers or prodrugs thereof, and optionally other drugs Mixtures of the active ingredients may contain other optional ingredients such as pharmaceutically acceptable carriers and/or excipients. The purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient and thereby exerts biological activity.

在本文中，用语“多个”包括两个或更多个，例如两个、三个、四个等。As used herein, the term "plurality" includes two or more, such as two, three, four, and the like.

本发明化合物的合成方法Method for synthesizing the compound of the present invention

为了完成本发明的目的，本发明采用如下技术方案：In order to accomplish the object of the present invention, the present invention adopts the following technical solutions:

本发明式(I)化合物的制备方法包括以下步骤：The preparation method of the compound of the formula (I) of the present invention comprises the following steps:

使式(Ia)化合物与R ³NH ₂反应，得到式(Ib)化合物；使式(Ib)化合物与式(Ic)化合物反应，得到式(IA)化合物；使式(IA)化合物与R ⁶H反应，得到式(I)化合物； Reaction of a compound of formula (Ia) with R ³ NH ₂ to give a compound of formula (Ib); reacting a compound of formula (Ib) with a compound of formula (Ic) to provide a compound of formula (IA); and a compound of formula (IA) with R ⁶ H reaction to give a compound of formula (I);

其中：among them:

X选自卤素，优选为氯或溴；X is selected from halogen, preferably chlorine or bromine;

R ⁶选自杂环基；且 R ^{6 is} selected from heterocyclic groups;

R ¹～R ⁵、R ⁷、X ¹和X ²的定义如式(I)中所述。 R ¹ to R ⁵ , R ⁷ , X ¹ and X ² are as defined in the formula (I).

附图说明DRAWINGS

图1为测试例2中WO2016139292的实施例1化合物、本发明实施例1和实施例4化合物对MV4-11荷瘤BALB/c裸鼠移植瘤平均肿瘤体积变化图。1 is a graph showing changes in mean tumor volume of MV4-11 tumor-bearing BALB/c nude mice xenografts of the compound of Example 1 of WO2016139292 in Test Example 2, and the compounds of Example 1 and Example 4 of the present invention.

图2为测试例2中WO2016139292的实施例1化合物、本发明实施例1和实施例4化合物对MV4-11荷瘤BALB/c裸鼠移植瘤平均相对肿瘤体积变化图。Figure 2 is a graph showing the mean relative tumor volume changes of the MV4-11 tumor-bearing BALB/c nude mice xenografts of the compound of Example 1 of WO2016139292 in Test Example 2, the compounds of Example 1 and Example 4 of the present invention.

具体实施方式Detailed ways

以下结合实施例用于进一步描述本发明，但这些实施例并非限制着本发明的范围。The invention is further described in the following examples, but these examples are not intended to limit the scope of the invention.

实施例Example

实施例给出了式(I)所表示的代表性化合物的制备及相关结构鉴定数据。必须说明，下述实施例是用于说明本发明而不是对本发明的限制。 ¹H NMR图谱是用Bruker仪器(400MHz)测定而得，化学位移用ppm表示。使用四甲基硅烷内标准(0.00ppm)。 ¹H NMR的表示方法：s＝单峰，d＝双重峰，t＝三重峰，m＝多重峰，br＝变宽的，dd＝双重峰的双重峰，dt＝三重峰的双重峰。若提供偶合常数时，其单位为Hz。 The examples give the preparation of representative compounds represented by formula (I) and related structural identification data. It is to be understood that the following examples are intended to illustrate the invention and not to limit the invention. ^{The 1} H NMR spectrum was measured using a Bruker instrument (400 MHz) and the chemical shift was expressed in ppm. The internal standard of tetramethylsilane (0.00 ppm) was used. ¹ H NMR representation: s = singlet, d = doublet, t = triplet, m = multiplet, br = broadened, dd = doublet of doublet, dt = doublet of triplet. If a coupling constant is provided, its unit is Hz.

质谱是用LC/MS仪测定得到，离子化方式可为ESI或APCI。Mass spectrometry was measured by LC/MS, and the ionization method was ESI or APCI.

薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板，薄层色谱法 (TLC)使用的硅胶板采用的规格是0.15mm～0.2mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。Thin layer chromatography silica gel plate uses Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate. The specification of silica gel plate used for thin layer chromatography (TLC) is 0.15mm~0.2mm. The specification for thin layer chromatography separation and purification is 0.4mm. ~0.5mm.

柱层析一般使用烟台黄海硅胶200～300目硅胶为载体。Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as a carrier.

在下列实施例中，除非另有指明，所有温度为摄氏温度，除非另有指明，各种起始原料和试剂来自市售或者是根据已知的方法合成，市售原料和试剂均不经进一步纯化直接使用，除非另有指明，市售厂家包括但不限于Aldrich Chemical Company，ABCR GmbH&Co.KG，Acros Organics，广赞化工科技有限公司和景颜化工科技有限公司等处购买。In the following examples, all temperatures are in degrees Celsius unless otherwise indicated, and unless otherwise indicated, the various starting materials and reagents are either commercially available or synthesized according to known methods, and the commercially available materials and reagents are not further processed. Purification is used directly, unless otherwise indicated, and commercially available, including but not limited to Aldrich Chemical Company, ABCR GmbH & Co. KG, Acros Organics, Guangzan Chemical Technology Co., Ltd. and Jingyan Chemical Technology Co., Ltd., etc.

CD ₃OD：氘代甲醇。 CD ₃ OD: Deuterated methanol.

CDCl ₃：氘代氯仿。 CDCl ₃ : deuterated chloroform.

DMSO-d ₆：氘代二甲基亚砜。 DMSO-d ₆ : deuterated dimethyl sulfoxide.

氩气氛是指反应瓶连接一个约1L容积的氩气气球。The argon atmosphere means that the reaction flask is connected to an argon balloon having a volume of about 1 L.

实施例中若无特殊说明，反应中的溶液是指水溶液。In the examples, the solution in the reaction means an aqueous solution unless otherwise specified.

对化合物进行纯化，采用硅胶柱层析和薄层色谱法，其中洗脱剂体系选自：A：石油醚和乙酸乙酯体系；B：二氯甲烷和甲醇体系；C：二氯甲烷：乙酸乙酯；其中溶剂的体积比根据化合物的极性不同而不同，也可以加入少量的酸性或碱性试剂进行调节，如醋酸或三乙胺等。The compound is purified by silica gel column chromatography and thin layer chromatography, wherein the eluent system is selected from the group consisting of: A: petroleum ether and ethyl acetate system; B: dichloromethane and methanol system; C: dichloromethane: acetic acid Ethyl ester; wherein the volume ratio of the solvent varies depending on the polarity of the compound, and may also be adjusted by adding a small amount of an acidic or alkaline agent such as acetic acid or triethylamine.

实施例1Example 1

5-(1-(1,3-二甲氧基丙-2-基)-7-氟-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮5-(1-(1,3-Dimethoxypropan-2-yl)-7-fluoro-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3-di Methylpyridine-2(1H)-one

第一步first step

2-(二苄基氨基)丙烷-1,3-二醇2-(dibenzylamino)propane-1,3-diol

将2-氨基丙烷-1,3-二醇1a(10.0g,109.7mmol)和碳酸钾(47.7g,345.6mmol)溶于300mL乙醇中，室温下搅拌，在反应液中滴加溴化苄(56g,329mmol)，回流反应12小时。过滤，将滤液减压浓缩，将得到的残留物溶于200mL二氯甲烷中，以水(100mL)洗涤，分去水层，有机相以无水硫酸钠干燥，减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：A体系)纯化，得到2-(二苄基氨基)丙烷-1,3-二醇1b(20g，白色固体)，产率：67％。2-Aminopropane-1,3-diol 1a (10.0 g, 109.7 mmol) and potassium carbonate (47.7 g, 345.6 mmol) were dissolved in 300 mL of ethanol, stirred at room temperature, and benzyl bromide was added dropwise to the reaction solution. 56 g, 329 mmol), refluxed for 12 hours. Filtration, the filtrate was concentrated under reduced pressure, and the residue was evaporated,jjjjjjjjjjjjjjjjjjjjj Purification by silica gel column chromatography (eluent: A) afforded 2-(dibenzylamino)propane-1,3-diol 1b (20 g, white solid).

¹H NMR(400MHz,CDCl ₃):δ7.40-7.25(m,10H),3.74(s,4H),3.69(m,2H),3.59(m,2H),2.99(m,3H). _{^{1 H NMR (400MHz, CDCl 3}} ): δ7.40-7.25 (m, 10H), 3.74 (s, 4H), 3.69 (m, 2H), 3.59 (m, 2H), 2.99 (m, 3H).

第二步Second step

N,N-二苄基-1,3-二甲氧基丙-2-胺N,N-dibenzyl-1,3-dimethoxypropan-2-amine

氩气保护下，将2-(二苄基氨基)丙烷-1,3-二醇1b(5g,18.4mmol)和氢氧化钾(8.3g,147.6mmol)溶于80mL二甲亚砜中，室温下搅拌10分钟。在反应液中滴加碘甲烷(10.4g,73.6mmol)，室温下反应1小时。在反应液中加入100mL水，淬灭反应。以乙酸乙酯(100mL×3)萃取，合并有机相，以饱和氯化钠水溶液(100mL×1)洗涤，以无水硫酸钠干燥，减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：A体系)纯化，得到N,N-二苄基-1,3-二甲氧基丙-2-胺1c(5g，白色固体)，产率：90.9％。2-(Dibenzylamino)propane-1,3-diol 1b (5 g, 18.4 mmol) and potassium hydroxide (8.3 g, 147.6 mmol) were dissolved in 80 mL of dimethyl sulfoxide under argon atmosphere at room temperature. Stir under 10 minutes. Methyl iodide (10.4 g, 73.6 mmol) was added dropwise to the reaction mixture, and the mixture was reacted at room temperature for 1 hour. 100 mL of water was added to the reaction solution to quench the reaction. The organic layer was extracted with EtOAc (EtOAc (EtOAc) (EtOAc. Purification of the eluent: System A) gave N,N-dibenzyl-1,3-dimethoxypropan-2-amine 1c (5 g, white solid), yield: 90.9%.

MS m/z(ESI)：300.0[M+1]MS m/z (ESI): 300.0 [M+1]

第三步third step

1,3-二甲氧基丙-2-胺1,3-dimethoxypropan-2-amine

氢气保护下，将N,N-二苄基-1,3-二甲氧基丙-2-胺1c(5g，16.7mmol)溶于50mL甲醇中，加入10％钯碳催化剂(500mg,10％)，置换氢气3次，室温下反 应12小时。过滤，将滤液减压浓缩，得到粗品1,3-二甲氧基丙-2-胺1d(1.99g，无色油状物)，产率：100％。N,N-dibenzyl-1,3-dimethoxypropan-2-amine 1c (5 g, 16.7 mmol) was dissolved in 50 mL of methanol under a hydrogen atmosphere, and a 10% palladium carbon catalyst (500 mg, 10%) was added. The hydrogen was replaced 3 times and reacted at room temperature for 12 hours. Filtration and concentration of the filtrate under reduced pressure afforded crude 1,3-dimethoxypropan-2-amine 1d (1.99 g, colorless oil).

MS m/z(ESI)：120.1[M+1]MS m/z (ESI): 120.1 [M+1]

¹H NMR(400MHz,CDCl ₃):δ5.72(s,2H),3.44(m,4H),3.32(m,1H)3.28(s,6H). _{^{1 H NMR (400MHz, CDCl 3}} ): δ5.72 (s, 2H), 3.44 (m, 4H), 3.32 (m, 1H) 3.28 (s, 6H).

第四步the fourth step

4-溴-N-(1,3-二甲氧基丙-2-基)-2-氟-6-硝基苯胺4-bromo-N-(1,3-dimethoxyprop-2-yl)-2-fluoro-6-nitroaniline

将5-溴-1,2-二氟-3-硝基苯1e(500mg,2.11mmol)和1,3-二甲氧基丙-2-胺1d(276mg,2.32mmol)溶于20mL乙腈中，加入碳酸钾(437mg,3.16mmol)，80℃下反应6小时。减压浓缩，除去溶剂，加入200mL水，以乙酸乙酯(300mL×3)萃取，合并有机相，以饱和氯化钠水溶液(100mL×1)洗涤，以无水硫酸钠干燥，减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：A体系)纯化，得到4-溴-N-(1,3-二甲氧基丙-2-基)-2-氟-6-硝基苯胺1f(560mg，黄色固体)，产率：78.7％。5-Bromo-1,2-difluoro-3-nitrobenzene 1e (500 mg, 2.11 mmol) and 1,3-dimethoxypropan-2-amine 1d (276 mg, 2.32 mmol) were dissolved in 20 mL acetonitrile Potassium carbonate (437 mg, 3.16 mmol) was added and the reaction was carried out at 80 ° C for 6 hours. After concentrating under reduced pressure, the solvent was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated. The residue obtained is purified by silica gel column chromatography (eluent: A system) to give 4-bromo-N-(1,3-dimethoxyprop-2-yl)-2-fluoro-6-nitrate Benzilide 1f (560 mg, yellow solid), yield: 78.7%.

MS m/z(ESI)：337.9[M+1]MS m/z (ESI): 337.9 [M+1]

第五步the fifth step

5-(5-溴-1-(1,3-二甲氧基丙-2-基)-7-氟-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮5-(5-Bromo-1-(1,3-dimethoxypropan-2-yl)-7-fluoro-1H-benzo[d]imidazol-2-yl)-1,3-dimethyl Pyridine-2(1H)-one

将4-溴-N-(1,3-二甲氧基丙-2-基)-2-氟-6-硝基苯胺1f(186mg,0.55mmol)和1,5-二甲基-6-氧代-1,6-二氢吡啶-3-甲醛1g(100mg,0.66mmol,根据专利申请WO2016146738(A1)公开的中间体2的制备方法制得)溶于5mL乙醇中，依次加入2mL水和连二亚硫酸钠(287mg,1.66mmol)，90℃下反应3小时。加入10mL水和10mL乙酸乙酯稀释反应液，以乙酸乙酯(10mL×3)萃取，合并有机相，以无水硫酸钠干燥，减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：B体系)纯化，得到5-(5-溴-1-(1,3-二甲氧基丙-2-基)-7-氟-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮1h(150mg，黄色固体)，产率：64.9％。4-Bromo-N-(1,3-dimethoxyprop-2-yl)-2-fluoro-6-nitroaniline 1f (186 mg, 0.55 mmol) and 1,5-dimethyl-6- Oxo-1,6-dihydropyridine-3-carbaldehyde 1 g (100 mg, 0.66 mmol, prepared according to the preparation method of intermediate 2 disclosed in patent application WO2016146738 (A1)) was dissolved in 5 mL of ethanol, and 2 mL of water was added in sequence. Sodium dithionite (287 mg, 1.66 mmol) was reacted at 90 ° C for 3 hours. The reaction mixture was diluted with EtOAc (EtOAc) (EtOAc. Deprotection: B system) purification, to give 5-(5-bromo-1-(1,3-dimethoxypropan-2-yl)-7-fluoro-1H-benzo[d]imidazol-2-yl - 1,3-Dimethylpyridine-2(1H)-one 1h (150 mg, yellow solid), yield: 64.9%.

MS m/z(ESI)：438.9[M+1]MS m/z (ESI): 438.9 [M+1]

第六步Step 6

5-(1-(1,3-二甲氧基丙-2-基)-7-氟-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮5-(1-(1,3-Dimethoxypropan-2-yl)-7-fluoro-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3-di Methylpyridine-2(1H)-one

氩气保护下，将5-(5-溴-1-(1,3-二甲氧基丙-2-基)-7-氟-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮1h(50mg,0.12mmol)、吗啉(21mg,0.24mmol)、三 (二亚苄基丙酮)二钯(11mg,0.012mmol)、2-二环己基磷-2,4,6-三异丙基联苯(11mg,0.024mmol)和叔丁醇钾(34mg,0.36mmol)溶于5mL1,4-二氧六环中，100℃下微波反应1小时。加入10mL水和10mL乙酸乙酯稀释反应液，以乙酸乙酯(10mL×3)萃取，合并有机相，以无水硫酸钠干燥，减压浓缩，得到的残留物用薄层色谱法(展开剂：B体系)纯化，得到5-(1-(1,3-二甲氧基丙-2-基)-7-氟-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮1(10mg，白色固体)，产率：18.5％。5-(5-Bromo-1-(1,3-dimethoxyprop-2-yl)-7-fluoro-1H-benzo[d]imidazol-2-yl)-1 under argon , 3-dimethylpyridine-2(1H)-one 1h (50mg, 0.12mmol), morpholine (21mg, 0.24mmol), tris(dibenzylideneacetone) dipalladium (11mg, 0.012mmol), 2- Dicyclohexylphosphorus-2,4,6-triisopropylbiphenyl (11 mg, 0.024 mmol) and potassium t-butoxide (34 mg, 0.36 mmol) were dissolved in 5 mL of 1,4-dioxane, microwave at 100 ° C Reaction for 1 hour. The reaction mixture was diluted with EtOAc (10 mL, EtOAc) :B system) purification to give 5-(1-(1,3-dimethoxypropan-2-yl)-7-fluoro-5-morpholinyl-1H-benzo[d]imidazol-2-yl )-1,3-Dimethylpyridine-2(1H)-one 1 (10 mg, white solid), yield: 18.5%.

MS m/z(ESI)：444.5[M+1]MS m/z (ESI): 444.5 [M+1]

¹H NMR(400MHz,CDCl ₃):δ7.98(s,1H),7.78(s,1H),7.07(s,1H),6.75-6.71(m,1H),4.88-4.83(m,1H),3.90-3.83(m,4H),3.80-3.75(m,4H),3.62(s,3H),3.26(s,6H),3.18-3.16(t,J＝4.0Hz,4H),2.22(s,3H). _{^{1 H NMR (400MHz, CDCl 3}} ): δ7.98 (s, 1H), 7.78 (s, 1H), 7.07 (s, 1H), 6.75-6.71 (m, 1H), 4.88-4.83 (m, 1H) , 3.90-3.83 (m, 4H), 3.80-3.75 (m, 4H), 3.62 (s, 3H), 3.26 (s, 6H), 3.18-3.16 (t, J = 4.0 Hz, 4H), 2.22 (s , 3H).

实施例2Example 2

5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮5-(7-fluoro-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridine -2(1H)-ketone

第一步first step

4-溴-2-氟-N-(1-甲氧基丁-2-基)-6-硝基苯胺4-bromo-2-fluoro-N-(1-methoxybutan-2-yl)-6-nitroaniline

将5-溴-1,2-二氟-3-硝基苯1e(600mg,2.52mmol)和1-甲氧基丁-2-胺2a(260mg,2.52mmol)溶于20mL乙腈中，加入碳酸钾(521mg,3.78mmol)，80℃下反应6小时。减压浓缩，除去溶剂，加入50mL水，以乙酸乙酯(30mL×3)萃取，合并有机相，以饱和氯化钠水溶液(100mL×1)洗涤，以无水硫酸钠干燥，减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：A体系)纯化，得到4-溴-2-氟-N-(1- 甲氧基丁-2-基)-6-硝基苯胺2b(600mg，黄色固体)，产率：74.2％。5-Bromo-1,2-difluoro-3-nitrobenzene 1e (600 mg, 2.52 mmol) and 1-methoxybutan-2-amine 2a (260 mg, 2.52 mmol) were dissolved in 20 mL of acetonitrile and carbonated. Potassium (521 mg, 3.78 mmol) was reacted at 80 ° C for 6 hours. After concentrating under reduced pressure, the solvent was evaporated, evaporated, evaporated, evaporated, evaporated, evaporated. The residue obtained is purified by silica gel column chromatography (eluent: A) to give 4-bromo-2-fluoro-N-(1-methoxybutan-2-yl)-6-nitroaniline 2b (600 mg, yellow solid), Yield: 74.2%.

MS m/z(ESI)：322.8[M+1]MS m/z (ESI): 322.8 [M+1]

第二步Second step

5-(5-溴-7-氟-1-(1-甲氧基丁-2-基)-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮5-(5-Bromo-7-fluoro-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridine-2 (1H)-ketone

将4-溴-2-氟-N-(1-甲氧基丁-2-基)-6-硝基苯胺2b(600mg,1.87mmol)和1,5-二甲基-6-氧代-1,6-二氢吡啶-3-甲醛1g(282mg,1.87mmol)溶于10mL乙醇中，依次加入2mL水和连二亚硫酸钠(813mg,4.67mmol)，100℃下反应3小时。减压浓缩，在残留物中加入50mL乙酸乙酯，搅拌，过滤，除去固体不溶物，将滤液减压浓缩，残留物溶解于甲苯(10mL×3)中进一步减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：A体系)纯化，得到5-(5-溴-7-氟-1-(1-甲氧基丁-2-基)-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮2c(650mg，黄色固体)，产率：82.2％。4-Bromo-2-fluoro-N-(1-methoxybutan-2-yl)-6-nitroaniline 2b (600 mg, 1.87 mmol) and 1,5-dimethyl-6-oxo- 1,6-dihydropyridine-3-carbaldehyde 1 g (282 mg, 1.87 mmol) was dissolved in 10 mL of ethanol, and 2 mL of water and sodium dithionite (813 mg, 4.67 mmol) were sequentially added, and the mixture was reacted at 100 ° C for 3 hours. The organic layer was concentrated under reduced pressure. EtOAc (EtOAc m. Purified by silica gel column chromatography (eluent: A system) to give 5-(5-bromo-7-fluoro-1-(1-methoxybut-2-yl)-1H-benzo[d]imidazole 2-yl)-1,3-dimethylpyridine-2(1H)-one 2c (650 mg, yellow solid), yield: 82.2%.

MS m/z(ESI)：421.8[M+1]MS m/z (ESI): 421.8 [M+1]

第三步third step

5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮5-(7-fluoro-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridine -2(1H)-ketone

氩气保护下，将5-(5-溴-7-氟-1-(1-甲氧基丁-2-基)-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮2c(220mg,0.52mmol)、吗啉(90.6mg,1.0mmol)、三(二亚苄基丙酮)二钯(47.6mg,0.052mmol)、2-二环己基磷-2,4,6-三异丙基联苯(47.6mg,0.1mmol)和叔丁醇钾(149.9mg,1.56mmol)溶于5mL1,4-二氧六环中，100℃下微波反应2小时。加入50mL水和30mL乙酸乙酯稀释反应液，以乙酸乙酯(30mL×2)萃取，合并有机相，以无水硫酸钠干燥，减压浓缩，得到的残留物用硅胶柱层析法(洗脱剂：B体系)纯化，得到5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮2(221mg，白色固体)，产率：99.1％。5-(5-Bromo-7-fluoro-1-(1-methoxybutan-2-yl)-1H-benzo[d]imidazol-2-yl)-1,3-protected under argon Lutidine-2(1H)-one 2c (220 mg, 0.52 mmol), morpholine (90.6 mg, 1.0 mmol), tris(dibenzylideneacetone)dipalladium (47.6 mg, 0.052 mmol), 2- Cyclohexylphosphorus-2,4,6-triisopropylbiphenyl (47.6 mg, 0.1 mmol) and potassium t-butoxide (149.9 mg, 1.56 mmol) were dissolved in 5 mL of 1,4-dioxane at 100 ° C The microwave was reacted for 2 hours. The reaction mixture was diluted with EtOAc (EtOAc) (EtOAc (EtOAc) Deprotection: B system) purification, to give 5-(7-fluoro-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl) -1,3-Dimethylpyridine-2(1H)-one 2 (221 mg, white solid), yield: 99.1%.

MS m/z(ESI)：429.0[M+1]MS m/z (ESI): 429.0 [M+1]

¹H NMR(400MHz,CDCl ₃)δ8.31(s,1H),7.80(s,1H),7.19(s,1H),6.81(d,J＝15.1Hz,1H),4.58(s,1H),4.05(t,J＝10.3Hz,1H),3.92-3.85(m,4H),3.81(d,J＝9.1Hz,1H),3.68(s,3H),3.36(s,3H),3.26-3.19(m,4H),2.23(s,3H),2.03-1.92(m,1H),1.90-1.79(m,1H),0.61(t,J＝7.3Hz,3H). _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.31 (s, 1H), 7.80 (s, 1H), 7.19 (s, 1H), 6.81 (d, J = 15.1Hz, 1H), 4.58 (s, 1H) , 4.05 (t, J = 10.3 Hz, 1H), 3.92-3.85 (m, 4H), 3.81 (d, J = 9.1 Hz, 1H), 3.68 (s, 3H), 3.36 (s, 3H), 3.26- 3.19 (m, 4H), 2.23 (s, 3H), 2.03-1.92 (m, 1H), 1.90 - 1.79 (m, 1H), 0.61 (t, J = 7.3 Hz, 3H).

实施例3和实施例4Example 3 and Example 4

(S)-5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基 吡啶-2(1H)-酮(S)-5-(7-fluoro-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3- Lutidine-2(1H)-one

(R)-5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮(R)-5-(7-fluoro-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3- Lutidine-2(1H)-one

第一步first step

(S)-5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮(S)-5-(7-fluoro-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3- Lutidine-2(1H)-one

(R)-5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮(R)-5-(7-fluoro-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3- Lutidine-2(1H)-one

将5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮2(210mg,0.49mmol)进一步通过采用超临界流体色谱(SFC)法，用高效液相制备色谱和手性柱对手性异构体进行拆分(手性柱ChiralPak AD,300×50mm I.D.,10μm；80mL/min；流动相为A(对于CO ₂)和B(对于ETOH)(0.1％NH ₃.H ₂O))，得到(S)-5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮3(57.09mg,白色固体)，产率：27.2％，100％ee，保留时间4.590min；(R)-5-(7-氟-1-(1-甲氧基丁-2-基)-5-吗啉基-1H-苯并[d]咪唑-2-基)-1,3-二甲基吡啶-2(1H)-酮4(60.38mg,白色固体)，产率：28.8％，100％ee，保留时间2.863min。 5-(7-Fluoro-1-(1-methoxybut-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethyl Pyridine-2(1H)-one 2 (210 mg, 0.49 mmol) was further resolved by supercritical fluid chromatography (SFC) using high performance liquid chromatography and chiral column chiral isomers (chiral Pak) AD, 300 × 50 mm ID, 10 μm; 80 mL / min; mobile phase A (for CO ₂ ) and B (for ETOH) (0.1% NH ₃ .H ₂ O)), (S)-5-(7- Fluor-1-(1-methoxybutan-2-yl)-5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridine-2(1H) -ketone 3 (57.09 mg, white solid), yield: 27.2%, 100% ee, retention time 4.590 min; (R)-5-(7-fluoro-1-(1-methoxybut-2-yl) -5-morpholinyl-1H-benzo[d]imidazol-2-yl)-1,3-dimethylpyridine-2(1H)-one 4 (60.38 mg, white solid), yield: 28.8 %, 100% ee, retention time 2.863min.

化合物3Compound 3

MS m/z(ESI)：429.0[M+1]MS m/z (ESI): 429.0 [M+1]

¹H NMR(400MHz,CDCl ₃)δ8.31(s,1H),7.80(s,1H),7.19(s,1H),6.81(d,J＝15.1Hz,1H),4.58(s,1H),4.05(t,J＝10.3Hz,1H),3.92-3.85(m,4H),3.81(d,J＝ 9.1Hz,1H),3.68(s,3H),3.36(s,3H),3.26-3.19(m,4H),2.23(s,3H),2.03-1.92(m,1H),1.90-1.79(m,1H),0.61(t,J＝7.3Hz,3H). _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.31 (s, 1H), 7.80 (s, 1H), 7.19 (s, 1H), 6.81 (d, J = 15.1Hz, 1H), 4.58 (s, 1H) , 4.05 (t, J = 10.3 Hz, 1H), 3.92-3.85 (m, 4H), 3.81 (d, J = 9.1 Hz, 1H), 3.68 (s, 3H), 3.36 (s, 3H), 3.26- 3.19 (m, 4H), 2.23 (s, 3H), 2.03-1.92 (m, 1H), 1.90 - 1.79 (m, 1H), 0.61 (t, J = 7.3 Hz, 3H).

化合物4Compound 4

MS m/z(ESI)：429.0[M+1]MS m/z (ESI): 429.0 [M+1]

¹H NMR(400MHz,CDCl ₃)δ8.31(s,1H),7.80(s,1H),7.19(s,1H),6.81(d,J＝15.1Hz,1H),4.58(s,1H),4.05(t,J＝10.3Hz,1H),3.92-3.85(m,4H),3.81(d,J＝9.1Hz,1H),3.68(s,3H),3.36(s,3H),3.26-3.19(m,4H),2.23(s,3H),2.03-1.92(m,1H),1.90-1.79(m,1H),0.61(t,J＝7.3Hz,3H). _{^{1 H NMR (400MHz, CDCl 3}} ) δ8.31 (s, 1H), 7.80 (s, 1H), 7.19 (s, 1H), 6.81 (d, J = 15.1Hz, 1H), 4.58 (s, 1H) , 4.05 (t, J = 10.3 Hz, 1H), 3.92-3.85 (m, 4H), 3.81 (d, J = 9.1 Hz, 1H), 3.68 (s, 3H), 3.36 (s, 3H), 3.26- 3.19 (m, 4H), 2.23 (s, 3H), 2.03-1.92 (m, 1H), 1.90 - 1.79 (m, 1H), 0.61 (t, J = 7.3 Hz, 3H).

生物学评价Biological evaluation

测试例1本发明化合物对BRD4蛋白活性测定Test Example 1 Determination of BRD4 Protein Activity by Compounds of the Invention

本发明的示例化合物通过以下方法测试其针对BRD4蛋白的生物活性。Exemplary compounds of the invention are tested for their biological activity against the BRD4 protein by the following method.

本测试方法采用Cisbio Assays公司的EPIGENEOUS ^TMBROMODOMAIN ASSAY在体外条件下，测试化合物对重组人源BRD4蛋白与乙酰化组蛋白多肽底物之间基于FRET(荧光能量共振转移)的相互作用的影响来表述化合物针对BRD4的生物活性。其中GST标记的重组人源BRD4(1/2)蛋白来源于BPS Bioscience，乙酰化组蛋白多肽底物[Lys(5,8,12,16)Ac]H4(1-21)-biotin peptide购于AnaSpec。 This test method uses Cisbio Assays EPIGENEOUS ^TM BROMODOMAIN ASSAY company under in vitro conditions, test compounds on recombinant human interaction effect on the FRET (fluorescence resonance energy transfer) to express protein and BRD4 between acetylated histone peptide substrate The biological activity of the compound against BRD4. The GST-tagged recombinant human BRD4(1/2) protein was derived from BPS Bioscience, and the acetylated histone polypeptide substrate [Lys(5,8,12,16)Ac]H4(1-21)-biotin peptide was purchased from AnaSpec.

具体实验方法和流程可参考EPIGENEOUS ^TMBROMODOMAIN ASSAY试剂盒说明书，实验流程简述如下： Specific experimental methods and procedures can refer to the EPIGENEOUS ^TM BROMODOMAIN ASSAY kit instructions, the experimental process is as follows:

本发明中的化合物先溶解于DMSO中，随后以试剂盒中提供的缓冲液将其稀释至测试所需浓度(终浓度范围10μM-0.1nM)。将2μL化合物加入到384孔微孔板中，随后分别加入4uL以缓冲液稀释的GST标记的重组人源BRD4(1/2)蛋白和4μL乙酰化组蛋白多肽底物，最后向孔中加入偶联有铕系元素化合物的抗GST抗体和偶联有链酶亲和素的FRET受体d2各5uL，振荡混匀后以封板膜密封，并在室温下振荡孵育3～5小时。化合物每个浓度设复孔对照，对照孔和空白孔中加入2μL DMSO。随后在兼容TF-FRET模式的酶标仪上测定读取各孔在对应铕系元素激发波长下，发射波长为620nM和665nM下的荧光强度。通过与对照组的荧光强度数值进行比较计算化合物在各浓度点下的抑制率，进而通过GraphPad Prism 5软件中以化合物浓度对数-抑制率进行非线性回归分析并得到化合物抑制BRD4蛋白与乙酰化组蛋白多肽底物相互作用的IC ₅₀值。 The compounds of the invention were first dissolved in DMSO and subsequently diluted to the desired concentration for testing (final concentration range 10 μM - 0.1 nM) in the buffer provided in the kit. 2 μL of the compound was added to a 384-well microtiter plate, followed by the addition of 4 uL of GST-labeled recombinant human BRD4 (1/2) protein and 4 μL of acetylated histone polypeptide substrate diluted in buffer, and finally added to the well. 5 μL of each of the anti-GST antibody linked to the lanthanide compound and the FRET receptor d2 conjugated with streptavidin, vortexed and sealed, sealed with a sealing membrane, and incubated at room temperature for 3 to 5 hours with shaking. For each concentration of the compounds, a duplicate well control was set, and 2 μL of DMSO was added to the control wells and blank wells. The fluorescence intensity at the emission wavelengths of 620 nM and 665 nM at the excitation wavelength of the corresponding lanthanide was then determined on a microplate reader compatible with the TF-FRET mode. The inhibition rate of the compound at each concentration point was calculated by comparing with the fluorescence intensity values of the control group, and then the nonlinear regression analysis of the compound concentration log-inhibition rate in GraphPad Prism 5 software was performed to obtain the compound inhibiting BRD4 protein and acetylation. IC ₅₀ values histone polypeptide substrate interaction.

表1本发明示例化合物对BRD4蛋白活性抑制的IC ₅₀数据 Table 1 IC ₅₀ data for inhibition of BRD4 protein activity by exemplary compounds of the invention

结论：本发明的示例化合物对于BRD4蛋白具有较好的抑制作用。Conclusion: The exemplified compounds of the present invention have a good inhibitory effect on the BRD4 protein.

备注：WO2016139292的实施例1化合物的结构式如下式所示：Remarks: The structural formula of the compound of Example 1 of WO2016139292 is as follows:

其制备方法参见WO2016139292的实施例1。See Method 1 of WO2016139292 for its preparation.

测试例2本发明化合物对***癌细胞(LNCaP)增殖抑制测定Test Example 2 Inhibition of proliferation of prostate cancer cells (LNCaP) by the compound of the present invention

本发明示例化合物的细胞水平活性通过CCK-8(Dojindo，东仁化学科技)以吸光度方法来检测化合物对细胞增殖的抑制作用。将处于对数生长期的***癌细胞LNCaP(购于中国科学院上海生命科学研究院细胞资源中心)以4000个每孔的密度接种至96孔培养板中，培养过夜后，加入不同浓度的测试化合物(终浓度浓度范围30μM～0.1nM)。在37℃，5％CO ₂条件下培养细胞72小时。培养结束后，向每孔加入适宜体积的CCK-8试剂(购于东仁化学科技(上海)有限公司，货号CK04)并在37℃下继续培养1～5小时，随后在酶标仪上读取各孔的在450nM下的吸光度数值。通过与对照组的吸光度数值进行比较计算化合物在各浓度点下的抑制率，进而通过GraphPad Prism 5软件中以化合物浓度对数-抑制率进行非线性回归分析并得到化合物抑制细胞增殖的IC ₅₀值。 The cell-level activity of the exemplified compounds of the present invention is measured by an absorbance method by CCK-8 (Dojindo, Toray Chemical Technology) to inhibit the proliferation of the compound. The prostate cancer cell LNCaP (purchased from the Shanghai Institute of Biological Sciences, Chinese Academy of Sciences) in the logarithmic growth phase was inoculated into a 96-well culture plate at a density of 4000 per well. After incubation overnight, different concentrations of test compounds were added. (The final concentration range is from 30 μM to 0.1 nM). The cells were cultured for 72 hours at 37 ° C, 5% CO ₂ . After the completion of the culture, a suitable volume of CCK-8 reagent (purchased from Dongren Chemical Technology (Shanghai) Co., Ltd., item number CK04) was added to each well and cultured at 37 ° C for 1 to 5 hours, followed by reading on the microplate reader. The absorbance values at 450 nM for each well were taken. The inhibition rate of the compound at each concentration point was calculated by comparison with the absorbance values of the control group, and then the nonlinear regression analysis of the compound concentration log-inhibition rate in GraphPad Prism 5 software was performed to obtain the IC ₅₀ value of the compound inhibiting cell proliferation. .

表2本发明示例化合物对LNCaP增殖抑制的IC ₅₀数据 Table Example Compound 2 of the present invention on proliferation of LNCaP IC ₅₀ data

结论：本发明的示例化合物对于***癌细胞LNCaP增殖具有较好的抑制作用。Conclusion: The exemplified compounds of the present invention have a good inhibitory effect on the proliferation of prostate cancer cells LNCaP.

测试例3本发明化合物对人类急性单核细胞白血病细胞MV4-11裸鼠移植瘤的生长抑制作用的测试Test Example 3 Test of the growth inhibitory effect of the compound of the present invention on human acute myeloid leukemia cell line MV4-11 nude mice xenografts

1.实验目的1. Experimental purpose

本测试用来评价连续22天，每天两次，口服给予实施例1、实施例4化合物以及WO2016139292的实施例1化合物，考察其对人类急性单核细胞白血病细胞MV4-11裸鼠移植瘤的生长抑制作用。This test was used to evaluate the compound of Example 1, the compound of Example 4 and the compound of Example 1 of WO2016139292 administered orally twice a day for 22 days to examine the growth of human MV4-11 xenografts in human acute monocytic leukemia cells. Inhibition.

2.受试物配制2. Preparation of test substance

2.1空白给药制剂配制2.1 blank drug preparation preparation

配制适量体积的含有90％PEG300和10％乙醇(10％TPGS，w/v)作为空白组给药试液。An appropriate volume of a test solution containing 90% PEG300 and 10% ethanol (10% TPGS, w/v) was prepared as a blank group.

2.2 WO2016139292的实施例1化合物给药制剂配制2.2 Example 1 of WO2016139292 for administration of a compound for administration

称取适量WO2016139292的实施例1化合物，加入适量体积含有90％PEG300和10％乙醇(10％TPGS，w/v)的溶媒，涡旋振荡均匀，配制成浓度为0.5mg/mL的给药制剂。An appropriate amount of the compound of Example 1 of WO2016139292 was weighed, and an appropriate amount of a solvent containing 90% PEG300 and 10% ethanol (10% TPGS, w/v) was added thereto, and the mixture was vortexed uniformly to prepare a preparation having a concentration of 0.5 mg/mL. .

2.3实施例1化合物口服给药制剂配制2.3 Example 1 compound oral administration preparation preparation

称取适量实施例1化合物，加入适量体积含有90％PEG300和10％乙醇(10％TPGS，w/v)的溶媒，涡旋振荡均匀，配制成浓度为0.5mg/mL的给药制剂。An appropriate amount of the compound of Example 1 was weighed, and an appropriate amount of a solvent containing 90% PEG300 and 10% ethanol (10% TPGS, w/v) was added thereto, and the mixture was vortexed uniformly to prepare a preparation having a concentration of 0.5 mg/mL.

2.4实施例4化合物口服给药制剂配制2.4 Example 4 compound oral administration preparation preparation

称取适量实施例4化合物，加入适量体积含有90％PEG300和10％乙醇(10％TPGS，w/v)的溶媒，涡旋振荡均匀，配制成浓度为0.5mg/mL的给药制剂。An appropriate amount of the compound of Example 4 was weighed, and an appropriate amount of a solvent containing 90% PEG300 and 10% ethanol (10% TPGS, w/v) was added thereto, and the mixture was vortexed uniformly to prepare a preparation having a concentration of 0.5 mg/mL.

3.实验动物3. Experimental animals

品种和品系：BALB/c裸鼠，SPF，雌性，6～7周龄(16～22克)，健康状况良好，32只，适应环境时间5～7天。合格证号：1140070017310，购买于北京维通利华实验动物技术有限公司。Varieties and strains: BALB/c nude mice, SPF, female, 6 to 7 weeks old (16 to 22 grams), healthy, 32, adapted to environmental time 5 to 7 days. Certificate No.: 1140070017310, purchased from Beijing Weitong Lihua Experimental Animal Technology Co., Ltd.

4. MV-4-11细胞培养4. MV-4-11 cell culture

在5％CO ₂、37℃培养条件下，MV4-11细胞(购买于美国模式培养物保藏所(ATCC))在含10％胎牛血清的IMDM培养基中进行常规细胞培养。以0.25％胰酶消化传代，根据细胞生长情况进行传代，传代比例为1:3或1:4。 MV4-11 cells (purchased from the American Type Culture Collection (ATCC)) were subjected to conventional cell culture in IMDM medium containing 10% fetal calf serum under 5% CO ₂ , 37 ° C culture conditions. Passage was digested with 0.25% trypsin and passaged according to cell growth conditions, with a passage ratio of 1:3 or 1:4.

5.动物接种及分组5. Animal vaccination and grouping

实验第0天，收取对数生长期MV4-11细胞，细胞计数后重悬于含50％的PBS(磷酸缓冲盐溶液)(pH7.4，0.01M)和50％的基质胶(Matrigel)中，调整细胞浓度至7.0×10 ⁷细胞/mL；将细胞置于冰盒中，用1-mL注射器吸取细胞悬液，注射到裸鼠前右侧腋窝皮下，每只动物接种200μL(14×106细胞/只)，建立MV4-11移植瘤模型。接种后第12天，肿瘤长至体积100～300mm ³左右时，选出肿瘤体积相近、形状较好的小鼠(形状尽量为单一圆球形，无不规则的形状或多个肿瘤聚在一起)每组8只，分为4组。 On day 0 of the experiment, MV4-11 cells in logarithmic growth phase were harvested, and the cells were counted and resuspended in 50% PBS (phosphate buffered saline) (pH 7.4, 0.01 M) and 50% matrigel (Matrigel). Adjust the cell concentration to 7.0×10 ⁷ cells/mL; place the cells in an ice box, aspirate the cell suspension with a 1-mL syringe, and inject into the right axilla of the nude mouse, inoculate 200 μL (14×106) per animal. Cells/only), established a MV4-11 xenograft model. On the 12th day after inoculation, when the tumor grows to a volume of about 100 to 300 mm ³ , mice with similar tumor volume and good shape are selected (the shape is as single a sphere as possible, no irregular shape or multiple tumors are gathered together) Group 8 was divided into 4 groups.

6.动物给药和观察6. Animal administration and observation

各组动物每天固定时间根据动物体重给予受试物1天1次(QD)，口服给药(po)，于分组当天(接种后第12天)，开始第一次给药，连续22天，并记录每天动物体重。Each group of animals was given a test substance once a day according to the body weight of the animal once a day (QD), orally (po), on the day of grouping (12 days after inoculation), the first administration was started for 22 consecutive days. And record the animal weight per day.

第1组灌胃给予空白溶剂，给药体积为10mL/kg，QD，PO；第2组灌胃给予WO2016139292的实施例1化合物，给药剂量5mg/kg，QD，PO；第3组给予实施例1化合物，给药剂量：第12～19天为5mg/kg，QD，PO；第20～33天为2mg/kg，QD，PO；第4组给予实施例4化合物，给药剂量：第12～19天为5mg/kg，QD，PO；第20～33天为2mg/kg，QD，PO。The first group was intragastrically administered with a blank solvent at a dose of 10 mL/kg, QD, PO; the second group was intragastrically administered with the compound of Example 1 of WO2016139292 at a dose of 5 mg/kg, QD, PO; Compound of Example 1, administered dose: 5 mg/kg on day 12-19, QD, PO; 2 mg/kg on day 20-33, QD, PO; Group 4 was administered compound of Example 4, dose: 5 to 19 days, 5 mg/kg, QD, PO; 20 to 33 days, 2 mg/kg, QD, PO.

观察各组动物接种部位肿瘤的形成状况，实验开始后每周测量2次瘤径，计算肿瘤体积，同时称量动物体重并记录。The formation of tumors at the inoculation site of each group of animals was observed. The tumor diameter was measured twice a week after the start of the experiment, the tumor volume was calculated, and the body weight of the animals was weighed and recorded.

肿瘤体积(TV)计算公式如下：The tumor volume (TV) is calculated as follows:

TV(mm ³)＝l×w ²/2 TV(mm ³ )=l×w ² /2

其中，l表示肿瘤长径(mm)；w表示肿瘤短径(mm)。Wherein, l represents the tumor long diameter (mm); w represents the tumor short diameter (mm).

抗肿瘤活性的评价指标：肿瘤生长抑制率TGI(％)，相对肿瘤增殖率T/C(％)。Evaluation index of antitumor activity: tumor growth inhibition rate TGI (%), relative tumor growth rate T/C (%).

相对肿瘤体积(RTV)的计算公式为：The relative tumor volume (RTV) is calculated as:

RTV＝100×TV _t/TV _initial RTV=100×TV _t /TV _initial

其中，TV _initial为分组时给药前测量到的肿瘤体积；TV _t为给药期间每一次测量时的肿瘤体积。 Wherein TV _initial is the tumor volume measured before administration at the time of grouping; TV _t is the tumor volume at each measurement during administration.

相对肿瘤增殖率T/C(％)的计算公式为：The relative tumor growth rate T/C (%) is calculated as:

T/C＝100％×(RTV _T/RTV _C) T/C=100%×(RTV _T /RTV _C )

其中，RTV _T表示治疗组RTV；RTV _C表示溶剂对照组RTV。 Among them, RTV _T represents the treatment group RTV; RTV _C represents the solvent control group RTV.

肿瘤生长抑制率TGI(％)的计算公式为：The formula for calculating the tumor growth inhibition rate TGI (%) is:

TGI＝100％×[1－(TV _t(T)－TV _initial(T))/(TV _t(C)－TV _initial(C))] TGI=100%×[1-(TV _t(T) -TV _initial(T) )/(TV _t(C) -TV _initial(C) )]

其中，TV _t(T)表示治疗组每次测量的肿瘤体积；TV _initial(T)表示分组时给药前治疗组的肿瘤体积；TV _t(C)表示溶剂对照组每次测量的肿瘤体积；TV _initial(C)表示分组时给药前溶剂对照组的肿瘤体积。 Where TV _t(T) represents the tumor volume measured each time in the treatment group; TV _{initial (T)} represents the tumor volume of the treatment group before administration in the group; TV _{t (C)} represents the tumor volume measured each time in the solvent control group; TV _{initial (C)} indicates the tumor volume of the solvent control group before administration at the time of grouping.

瘤重抑瘤率IR(％)的计算公式为：The formula for calculating the tumor weight inhibition rate IR (%) is:

IR＝100％×(W _C－W _T)/W _C IR=100%×(W _C -W _T )/W _C

其中，W _C表示对照组瘤重；W _T表示治疗组瘤重。 Wherein, W _C represents the tumor weight of the control group; W _T represents the tumor weight of the treatment group.

7.结果7. Results

WO2016139292的实施例1化合物、本发明实施例1和实施例4化合物对MV4-11荷瘤BALB/c裸鼠移植瘤平均肿瘤体积变化图见图1；The average tumor volume change of the compound of Example 1 of WO2016139292, the compound of Example 1 and the compound of Example 4 for the transplanted tumor of MV4-11 tumor-bearing BALB/c nude mice is shown in Fig. 1;

WO2016139292的实施例1化合物、本发明实施例1和实施例4化合物对MV4-11荷瘤BALB/c裸鼠移植瘤平均相对肿瘤体积变化图见图2。The average relative tumor volume change of the compound of Example 1 of WO2016139292, the compound of Example 1 and Example 4 of the present invention for MV4-11 tumor-bearing BALB/c nude mice is shown in Fig. 2.

表3本发明化合物对MV4-11荷瘤BALB/c裸鼠移植瘤的生长抑制率(TGI％)的影响Table 3 Effect of the compounds of the present invention on the growth inhibition rate (TGI%) of MV4-11 tumor-bearing BALB/c nude mice xenografts

表4本发明化合物对MV4-11荷瘤BALB/c裸鼠移植瘤的相对肿瘤增殖率T/C(％)的影响Table 4 Effect of the compound of the present invention on the relative tumor growth rate T/C(%) of MV4-11 tumor-bearing BALB/c nude mice xenografts

表5.实验结束时各组动物瘤重及瘤重抑瘤率Table 5. Tumor weight and tumor weight inhibition rate of each group at the end of the experiment

由表3～5、图1～2可知，在5mg/kg；2mg/kg(PO，QD)剂量下，本发明实施例1和4化合物在给药22天内对MV-411细胞建立小鼠体内肿瘤模型具有明显的生长抑制作用，且两者的活性均优于WO2016139292的实施例1的化合物。It can be seen from Tables 3 to 5 and Figures 1 to 2 that the compounds of Examples 1 and 4 of the present invention establish mice in MV-411 cells within 22 days of administration at a dose of 5 mg/kg; 2 mg/kg (PO, QD). The tumor model has significant growth inhibition and both have better activity than the compound of Example 1 of WO2016139292.

在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。All documents mentioned in the present application are hereby incorporated by reference in their entirety in their entireties in the the the the the the the the the In addition, it should be understood that various modifications and changes may be made by those skilled in the art in the form of the present invention.

Claims (17)

1. 式(I)所示的化合物：a compound of formula (I):

   

   

   包括其立体异构体、互变异构体或其可药用的盐，Including stereoisomers, tautomers or pharmaceutically acceptable salts thereof,

   其中：among them:

   X ¹选自-CH＝或-N＝； X ^{1 is} selected from -CH= or -N=;

   X ²选自-CH＝或-N＝； X ^{2 is} selected from -CH= or -N=;

   且X ¹和X ²不同时为-N＝； And X ¹ and X ^{2 are} not -N=;

   R ¹选自烷基；优选为甲基； R ^{1 is} selected from an alkyl group; preferably a methyl group;

   R ²选自氢原子或烷基；优选为甲基； R ^{2 is} selected from a hydrogen atom or an alkyl group; preferably a methyl group;

   R ³选自烷基，其中所述的烷基进一步被一个或多个烷氧基所取代； R ^{3 is} selected from alkyl, wherein said alkyl group is further substituted with one or more alkoxy groups;

   R ⁴选自卤素； R ^{4 is} selected from halogen;

   R ⁵选自氢原子、烷基、氰基、卤素、烷氧基、环烷基、杂环基、芳基、杂芳基、-OR ¹⁰、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹，其中所述的烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹的取代基所取代； R ^{5 is} selected from the group consisting of a hydrogen atom, an alkyl group, a cyano group, a halogen, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, -OR ¹⁰ , -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , —C(O)R ¹⁰ , —C(O)OR ¹⁰ or —NR ⁸ C(O)R ⁹ , wherein said alkyl group, cycloalkyl group, heterocyclic group, aryl group or heteroaryl group Further optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -NR ⁸ R ^{9, -C (O) NR 8} R 9, -C (O) R 10, -C (O) oR 10 , or -NR ⁸ C (O) R ⁹ is substituted with a substituent;

   R ⁶选自氢原子、烷基、氰基、烷氧基、环烷基、杂环基、芳基、杂芳基、-OR ¹⁰、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹，其中所述的烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ⁸R ⁹、-C(O)NR ⁸R ⁹、-C(O)R ¹⁰、-C(O)OR ¹⁰或-NR ⁸C(O)R ⁹的取代基所取代； R ^{6 is} selected from the group consisting of a hydrogen atom, an alkyl group, a cyano group, an alkoxy group, a cycloalkyl group, a heterocyclic group, an aryl group, a heteroaryl group, -OR ¹⁰ , -NR ⁸ R ⁹ , -C(O)NR ⁸ R ⁹ , —C(O)R ¹⁰ , —C(O)OR ¹⁰ or —NR ⁸ C(O)R ⁹ , wherein said alkyl, cycloalkyl, heterocyclic, aryl or heteroaryl is Further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, -NR ⁸ R ⁹ , Substituted by a substituent of -C(O)NR ⁸ R ⁹ , -C(O)R ¹⁰ , -C(O)OR ¹⁰ or -NR ⁸ C(O)R ⁹ ;

   R ⁷选自氢原子或烷基；优选为氢原子； R ^{7 is} selected from a hydrogen atom or an alkyl group; preferably a hydrogen atom;

   R ⁸、R ⁹和R ¹⁰各自独立地选自氢原子、烷基、环烷基、杂环基、芳基或杂芳基，其中所述烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ¹¹R ¹²、-C(O)NR ¹¹R ¹²、-C(O)R ¹³、-C(O)OR ¹³或-NR ¹¹C(O)R ¹²的取代基所取代； R ⁸ , R ⁹ and R ¹⁰ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group Or a heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0, - Substituted by a substituent of NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

   或者，R ⁸和R ⁹与相连接的N原子一起形成一个4～8元杂环基，其中4～8元杂环内含有一个或多个N、O、S(O) _p原子，并且4～8元杂环上任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、＝O、-NR ¹¹R ¹²、-C(O)NR ¹¹R ¹²、-C(O)R ¹³、-C(O)OR ¹³或-NR ¹¹C(O)R ¹²的取代基所取代； Alternatively, R ⁸ and R ⁹ together with the N atom to which they are attached form a 4 to 8 membered heterocyclic group, wherein the 4 to 8 membered heterocyclic ring contains one or more N, O, S(O) _p atoms, and 4 Optionally, one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, =0 Substituting a substituent of -NR ¹¹ R ¹² , -C(O)NR ¹¹ R ¹² , -C(O)R ¹³ , -C(O)OR ¹³ or -NR ¹¹ C(O)R ¹² ;

   R ¹¹、R ¹²和R ¹³各自独立地选自氢原子、烷基、环烷基、杂环基、芳基或杂芳基，其中所述烷基、环烷基、杂环基、芳基或杂芳基任选进一步被一个或多个选自羟基、卤素、硝基、氰基、烷基、烷氧基、环烷基、杂环基、芳基、杂芳基、羧基或羧酸酯基的取代基所取代；且 R ¹¹ , R ¹² and R ¹³ are each independently selected from a hydrogen atom, an alkyl group, a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group Or a heteroaryl group optionally further selected from one or more selected from the group consisting of hydroxyl, halogen, nitro, cyano, alkyl, alkoxy, cycloalkyl, heterocyclyl, aryl, heteroaryl, carboxy or carboxylic acid Substituted by a substituent of an ester group;

   p为0、1或2。p is 0, 1, or 2.
2. 根据权利要求1所述的化合物，其具有式(II)所示结构：The compound according to claim 1, which has the structure represented by the formula (II):

   

   

   其中：among them:

   R ^a和R ^b各自独立地选自氢原子或烷氧基，且R ^a和R ^b不同时为氢原子；所述的烷氧基优选为甲氧基； R ^a and R ^b are each independently selected from a hydrogen atom or an alkoxy group, and R ^a and R ^{b are} not simultaneously a hydrogen atom; the alkoxy group is preferably a methoxy group;

   m为1、2、3或4；优选为1或2；m is 1, 2, 3 or 4; preferably 1 or 2;

   n为1、2、3或4；优选为1或2；且n is 1, 2, 3 or 4; preferably 1 or 2;

   R ¹、R ²、R ⁴～R ⁷的定义如权利要求1中所述。 R ¹ , R ² , R ⁴ to R ⁷ are as defined in claim 1.
3. 根据权利要求2所述的化合物，其具有式(III)所示结构：The compound according to claim 2 having the structure represented by formula (III):

   

   

   其中：among them:

   R ^a为氢原子； R ^a is a hydrogen atom;

   R ^b选自烷氧基，优选为甲氧基； R ^{b is} selected from alkoxy groups, preferably methoxy groups;

   m为1、2、3或4；优选为1或2；m is 1, 2, 3 or 4; preferably 1 or 2;

   n为1、2、3或4；优选为1或2；且n is 1, 2, 3 or 4; preferably 1 or 2;

   R ¹、R ²、R ⁴～R ⁷的定义如权利要求1中所述。 R ¹ , R ² , R ⁴ to R ⁷ are as defined in claim 1.
4. 根据权利要求2所述的化合物，其具有式(IV)所示结构：The compound according to claim 2 having the structure represented by the formula (IV):

   

   

   其中：among them:

   R ^a为氢原子； R ^a is a hydrogen atom;

   R ^b选自烷氧基，优选为甲氧基； R ^{b is} selected from alkoxy groups, preferably methoxy groups;

   m为1、2、3或4；优选为1或2；m is 1, 2, 3 or 4; preferably 1 or 2;

   n为1、2、3或4；优选为1或2；且n is 1, 2, 3 or 4; preferably 1 or 2;

   R ¹、R ²、R ⁴～R ⁷的定义如权利要求1中所述。 R ¹ , R ² , R ⁴ to R ⁷ are as defined in claim 1.
5. 根据权利要求1～4中任一项所述的化合物，其中R ⁴选自氟、氯或溴；优选为氟。 The compound according to any one of claims 1 to 4, wherein R ^{4 is} selected from fluorine, chlorine or bromine; preferably fluorine.
6. 根据权利要求1～4中任一项所述的化合物，其中R ⁶选自杂环基，优选为吗啉基。 The compound according to any one of claims 1 to 4, wherein R ^{6 is} selected from a heterocyclic group, preferably a morpholinyl group.
7. 根据权利要求1所述的化合物，其中所述的化合物选自：The compound of claim 1 wherein said compound is selected from the group consisting of:

   

   
8. 根据权利要求1所述的式(I)化合物的制备方法，所述方法包括：A process for the preparation of a compound of formula (I) according to claim 1 comprising:

   

   

   使式(IA)化合物与R ⁶H反应，得到式(I)化合物； Reaction of a compound of formula (IA) with R ⁶ H to provide a compound of formula (I);

   其中：among them:

   X选自卤素，优选为氯或溴；X is selected from halogen, preferably chlorine or bromine;

   R ⁶选自杂环基；且 R ^{6 is} selected from heterocyclic groups;

   R ¹～R ⁵、R ⁷、X ¹和X ²的定义如权利要求1中所述。 R ¹ to R ⁵ , R ⁷ , X ¹ and X ² are as defined in claim 1.
9. 式(IA)所示的化合物：a compound of the formula (IA):

   

   

   其中：among them:

   X选自卤素，优选为氯或溴；且X is selected from halogen, preferably chlorine or bromine;

   R ¹～R ⁵、R ⁷、X ¹和X ²的定义如权利要求1中所述。 R ¹ to R ⁵ , R ⁷ , X ¹ and X ² are as defined in claim 1.
10. 根据权利要求9所述的化合物，其中所述的化合物选自：The compound of claim 9 wherein said compound is selected from the group consisting of:

    

    
11. 根据权利要求9所述的式(IA)化合物的制备方法，所述方法包括：A method of preparing a compound of formula (IA) according to claim 9, the method comprising:

    

    

    使式(Ib)化合物与式(Ic)化合物反应，得到式(IA)化合物；Reaction of a compound of formula (Ib) with a compound of formula (Ic) to provide a compound of formula (IA);

    其中：among them:

    X选自卤素，优选为氯或溴；且X is selected from halogen, preferably chlorine or bromine;

    R ¹～R ⁵、R ⁷、X ¹和X ²的定义如权利要求1中所述。 R ¹ to R ⁵ , R ⁷ , X ¹ and X ² are as defined in claim 1.
12. 一种药物组合物，所述的药物组合物含有有效剂量的根据权利要求1～7中任一项所述的化合物，及可药用的载体、赋形剂或它们的组合。A pharmaceutical composition comprising an effective amount of a compound according to any one of claims 1 to 7, together with a pharmaceutically acceptable carrier, excipient or a combination thereof.
13. 根据权利要求1～7中任一项所述的化合物，或根据权利要求12所述的药物组合物在制备用作溴结构域蛋白抑制剂的药物中的用途，其中所述的溴结构域蛋白优选为BRD2、BRD3和BRD4，更优选为BRD4。The use of a compound according to any one of claims 1 to 7, or a pharmaceutical composition according to claim 12, for the preparation of a medicament for use as a bromodomain protein inhibitor, wherein said bromodomain protein Preferred are BRD2, BRD3 and BRD4, more preferably BRD4.
14. 根据权利要求1～7中任一项所述的化合物或根据权利要求12所述的药物组合物在制备用于治疗或预防与溴结构域蛋白相关的疾病的药物中的用途，其中所述的疾病优选为癌症或炎症，其中所述的炎症优选为类风湿性关节炎、克隆恩病、湿疹、巨细胞性动脉炎、肝炎、炎性肠病、骨关节炎、胰腺炎、肺炎、银屑病、银屑病性关节炎、***性红斑狼疮、肾小球性肾炎、狼疮性肾炎、膜性肾小球肾炎或心肌炎；所述的炎症更优选为类风湿性关节炎。Use of a compound according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 12 for the manufacture of a medicament for the treatment or prevention of a disease associated with a bromodomain protein, wherein said The disease is preferably cancer or inflammation, wherein the inflammation is preferably rheumatoid arthritis, cloning, eczema, giant cell arteritis, hepatitis, inflammatory bowel disease, osteoarthritis, pancreatitis, pneumonia, psoriasis Disease, psoriatic arthritis, systemic lupus erythematosus, glomerulonephritis, lupus nephritis, membranous glomerulonephritis or myocarditis; the inflammation is more preferably rheumatoid arthritis.
15. 根据权利要求14所述的用途，其中所述的癌症为小细胞肺癌、非小细胞肺癌、乳腺癌、结直肠癌、***癌、黑色素瘤、胰腺癌、神经胶质瘤、脑瘤、***、卵巢癌、胰腺癌、***癌、肾细胞癌、胃癌、膀胱癌、肝癌、睾丸核蛋白中线癌、多发性骨髓瘤、急性髓性白血病、急性淋巴细胞性白血病、慢性淋巴细胞性白血病、慢性髓细胞性白血病或慢性骨髓性白血病。The use according to claim 14, wherein the cancer is small cell lung cancer, non-small cell lung cancer, breast cancer, colorectal cancer, prostate cancer, melanoma, pancreatic cancer, glioma, brain tumor, cervical cancer. , ovarian cancer, pancreatic cancer, prostate cancer, renal cell carcinoma, gastric cancer, bladder cancer, liver cancer, testicular nucleoprotein, midline cancer, multiple myeloma, acute myeloid leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, chronic Myeloid leukemia or chronic myelogenous leukemia.
16. 根据权利要求1～7中任一项所述的化合物或根据权利要求12所述的药物组合物在制备治疗或预防糖尿病性肾病、高血压性肾病、HIV-相关的肾病、多囊性肾病、肥胖、血脂异常、高胆固醇血症、阿尔茨海默病、代谢综合征、脂肪肝、II型糖尿病、胰岛素抵抗、糖尿病性视网膜病或糖尿病性神经病的药物中的用途。The compound according to any one of claims 1 to 7 or the pharmaceutical composition according to claim 12 for the preparation or treatment of diabetic nephropathy, hypertensive nephropathy, HIV-related nephropathy, polycystic kidney disease, Use in obesity, dyslipidemia, hypercholesterolemia, Alzheimer's disease, metabolic syndrome, fatty liver, type 2 diabetes, insulin resistance, diabetic retinopathy or diabetic neuropathy.
17. 治疗或预防与溴结构域蛋白相关的疾病的方法，包括向有此需要的对象施用根据权利要求1至7中任一项所述的化合物或根据权利要求12所述的药物组合物。A method of treating or preventing a disease associated with a bromodomain protein, comprising administering a compound according to any one of claims 1 to 7 or a pharmaceutical composition according to claim 12 to a subject in need thereof.

Images