CN108530337B - Indoleamide compound capable of selectively inhibiting gastric cancer cells - Google Patents

Abstract

Translated fromChinese

本发明公开了一种可选择性抑制胃癌细胞的吲哚酰胺类化合物，其能够选择性抑制胃癌细胞，尤其是选择性抑制MGC‑803(人胃癌细胞)细胞株。因而，可以用做选择性治疗胃癌的药物，具有良好的开发前景。The invention discloses an indoleamide compound that can selectively inhibit gastric cancer cells, which can selectively inhibit gastric cancer cells, especially MGC-803 (human gastric cancer cell) cell line. Therefore, it can be used as a drug for selective treatment of gastric cancer, and has a good development prospect.

Description

Translated fromChinese

一种可选择性抑制胃癌细胞的吲哚酰胺类化合物An indoleamide compound that selectively inhibits gastric cancer cells

技术领域technical field

本发明涉及药物化学领域，具体涉及一系列吲哚酰胺类化合物、其制备方法及其用途。The invention relates to the field of medicinal chemistry, in particular to a series of indole amide compounds, a preparation method thereof and uses thereof.

背景技术Background technique

胃癌是发生在胃上皮组织的恶性肿瘤，在我国其发病率居各类肿瘤的首位。在胃癌发生和演变的多步骤转化过程中细胞凋亡调节系统发生紊乱，是细胞表现出恶性表型的重要原因之一。Gastric cancer is a malignant tumor that occurs in gastric epithelial tissue, and its incidence ranks first among all types of tumors in my country. During the multi-step transformation process of gastric carcinogenesis and evolution, the disorder of the apoptosis regulatory system is one of the important reasons for the malignant phenotype of cells.

吲哚类化合物是一类重要的杂环化合物，具有广泛的生物活性。十字花科蔬菜中及大量海洋生物和放线菌中广泛存在着吲哚类次生代谢产物。近年来，其在抗癌方面的活性引起了人们的普遍关注。目前，SU11248(商品名：舒尼替尼)、长春碱(VLB，Vinblastine)、长春新碱(VCR，Vincristine)、长春地辛(VDS，Vindesine)、长春瑞滨(VBR，Vinorelbine)、靛玉红等少量含吲哚结构的品种已上市投入使用，毒副作用小和选择性强等特点已凸现出吲哚类抗癌化合物的特殊效果(刘小余，欧阳贵平.吲哚类抗癌化合物的研究进展,精细化工中间体,2010年10月第40卷第5期1-8页)。李雪琳等（CN103214472A）报道了一类丙烯酸吲哚酰胺类化合物

，该化合物对于肺癌、乳腺癌、骨肉瘤、宫颈癌等多种癌细胞具有抑制活性。宋军,等（《华中科技大学学报（医学版）》，2004年，第33卷，第6期，第720-723页）报道了吲哚美幸可诱导胃癌细胞凋亡，但是其机制不明确。Indoles are an important class of heterocyclic compounds with a wide range of biological activities. Indole secondary metabolites are widely found in cruciferous vegetables and a large number of marine organisms and actinomycetes. In recent years, its anticancer activity has attracted widespread attention. Currently, SU11248 (trade name: sunitinib), vinblastine (VLB, Vinblastine), vincristine (VCR, Vincristine), vindesine (VDS, Vindesine), vinorelbine (VBR, Vinorelbine), indigo A small amount of varieties containing indole structures such as red have been put into use, and the characteristics of small toxic side effects and strong selectivity have highlighted the special effects of indole anticancer compounds (Liu Xiaoyu, Ouyang Guiping. Research on indole anticancer compounds Progress, Fine Chemical Intermediates, October 2010, Vol. 40, No. 5, pp. 1-8). Li Xuelin et al. (CN103214472A) reported a class of acrylic acid indole amides

, the compound has inhibitory activity against lung cancer, breast cancer, osteosarcoma, cervical cancer and other cancer cells. Song Jun, et al. (Journal of Huazhong University of Science and Technology (Medical Edition), 2004, Vol. 33, No. 6, pp. 720-723) reported that indomethacin can induce apoptosis in gastric cancer cells, but the mechanism is unclear .

随着现代医学的发展，对于疾病进行选择性和/或靶向治疗是新药研发人员越来越关注的一个方向。因为，作为选择性和/或靶向药物，其能针对特定的病症、病理部位进行作用，进而减少副作用，节约用药成本。现有技术中虽然报道了一系列可用于治疗胃癌的吲哚类化合物，但是所述化合物通常同时针对多种癌细胞具有抑制作用，较少具有选择性抑制胃癌的药物。基于现有技术中的上述缺陷，开发可用于选择性抑制胃癌细胞的药物，对于选择性治疗胃癌具有重大意义。With the development of modern medicine, selective and/or targeted treatment of diseases is a direction that new drug developers pay more and more attention to. Because, as a selective and/or targeted drug, it can act on specific conditions and pathological sites, thereby reducing side effects and saving drug costs. Although a series of indole compounds that can be used for the treatment of gastric cancer have been reported in the prior art, the compounds usually have inhibitory effects on a variety of cancer cells at the same time, and there are few drugs that selectively inhibit gastric cancer. Based on the above-mentioned defects in the prior art, the development of drugs that can be used to selectively inhibit gastric cancer cells is of great significance for the selective treatment of gastric cancer.

发明内容SUMMARY OF THE INVENTION

本发明所要解决的技术问题是：提供了一类吲哚酰胺类化合物，其可选择性治疗胃癌，尤其是选择性抑制胃癌MGC-803细胞株，具有较好的开发应用前景。The technical problem to be solved by the present invention is: to provide a class of indoleamide compounds, which can selectively treat gastric cancer, especially selectively inhibit gastric cancer MGC-803 cell line, and have good development and application prospects.

本发明的第一个方面，是提供一类通式I化合物及其药学上可接受的盐，所示化合物可选择性抑制胃癌细胞的生长。The first aspect of the present invention is to provide a class of compounds of general formula I and pharmaceutically acceptable salts thereof, which can selectively inhibit the growth of gastric cancer cells.

其中：R各自独立地选自卤素、-NO2、-CN、C1-C8烷基、C1-C8烷氧基、C1-C8卤代烷基；Wherein: R is independently selected from halogen, -NO2, -CN, C1-C8 alkyl, C1-C8 alkoxy, C1-C8 haloalkyl;

n选自1、2、3或4。n is selected from 1, 2, 3 or 4.

优选地，R各自独立地选自卤素、C1-C4烷基、C1-C4烷氧基、C1-C4卤代烷基；Preferably, each R is independently selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl;

更优选地，R各自独立地选自氟、氯、溴、甲基、乙基、甲氧基、乙氧基、三氟甲基；More preferably, each R is independently selected from fluorine, chlorine, bromine, methyl, ethyl, methoxy, ethoxy, trifluoromethyl;

最优选地，R各自独立地选自甲基、氯、溴。Most preferably, each R is independently selected from methyl, chloro, bromo.

优选地，n选自1或2；Preferably, n is selected from 1 or 2;

更优选地，n为1。More preferably, n is 1.

优选地，本发明化合物选自化合物R2、R3、R4、R5或R6，其分别具有如下结构：Preferably, the compound of the present invention is selected from compounds R2, R3, R4, R5 or R6, which respectively have the following structures:

R2R2

R3 R4R3 R4

R5 R6R5 R6

本发明的另一方面提供一种制备式I化合物的方法，其合成路线如下：Another aspect of the present invention provides a kind of method for preparing the compound of formula I, and its synthetic route is as follows:

；

;

其中，R和n的选择范围如前所述。Among them, the selection range of R and n is as described above.

具体反应步骤如下：The specific reaction steps are as follows:

步骤一：在反应容器中加入苯胺和盐酸，将亚硝酸钠溶液滴加到溶液中。反应1小时抽滤，滤液滴加至亚硫酸钠溶液中，80℃反应2小时，再滴加浓盐酸，95℃反应2小时后置于常温过夜，析出苯肼盐酸盐固体。Step 1: Add aniline and hydrochloric acid to the reaction vessel, and drop the sodium nitrite solution into the solution. The reaction was carried out for 1 hour by suction filtration, the filtrate was added dropwise to sodium sulfite solution, reacted at 80°C for 2 hours, then concentrated hydrochloric acid was added dropwise, reacted at 95°C for 2 hours, and then left at room temperature overnight to precipitate solid phenylhydrazine hydrochloride.

步骤二：在反应容器中依次加入苯肼盐酸盐，4-氯-1-羟基-丁烷磺酸钠，乙醇，水，再调节溶液PH至6-7，75℃反应6小时；减压蒸馏除去乙醇及部分水，剩余溶液先用二氯甲烷洗涤，再调节PH至8-10，再用氯仿洗涤，置于4℃冰箱冷却过夜，析出色胺盐酸盐固体。Step 2: Add phenylhydrazine hydrochloride, sodium 4-chloro-1-hydroxy-butane sulfonate, ethanol, water in turn in the reaction vessel, then adjust the pH of the solution to 6-7, and react at 75°C for 6 hours; reduce the pressure Ethanol and part of water were distilled off, the remaining solution was washed with dichloromethane first, then adjusted to pH 8-10, washed with chloroform, placed in a 4°C refrigerator to cool overnight, and the solid of tryptamine hydrochloride was precipitated.

步骤三：在反应容器中依次加入水杨酸衍生物，1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐，1-羟基苯并三唑，丙酮，常温搅拌2小时；再加入步骤二所得色胺盐酸盐，无水碳酸钾，常温搅拌至反应完全；柱色谱纯化，得式I所示化合物。Step 3: Add salicylic acid derivatives, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride, 1-hydroxybenzotriazole, acetone, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, 1-hydroxybenzotriazole, acetone, and Stir for 2 hours; then add tryptamine hydrochloride obtained in step 2, anhydrous potassium carbonate, stir at room temperature until the reaction is complete; purify by column chromatography to obtain the compound shown in formula I.

其中，步骤一的苯胺、亚硝酸钠和亚硫酸钠的摩尔比为1 : 1-1.5 : 2-4.5，优选为1 : 1.05 : 3；Wherein, the mol ratio of aniline, sodium nitrite and sodium sulfite of step 1 is 1: 1-1.5: 2-4.5, preferably 1: 1.05: 3;

步骤二的苯肼盐酸盐和4-氯-1-羟基-丁烷磺酸钠的摩尔比为1 : 1-1.5 ，优选为1 : 1-1.2；The mol ratio of the phenylhydrazine hydrochloride of step 2 and sodium 4-chloro-1-hydroxy-butanesulfonate is 1: 1-1.5, preferably 1: 1-1.2;

步骤三的水杨酸衍生物、1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐、1-羟基苯并三唑和色胺盐酸盐的摩尔比为1 : 1-2 : 1-2 : 0.5-1.5；优选为1 : 1-1.5 : 1-1.5: 0.8-1.2。The mol ratio of the salicylic acid derivative of step 3, 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride, 1-hydroxybenzotriazole and tryptamine hydrochloride is: 1 : 1-2 : 1-2 : 0.5-1.5; preferably 1 : 1-1.5 : 1-1.5: 0.8-1.2.

本发明的另一方面提供一种药物组合物，其包含式I所示的化合物或其药学上可接受的盐，以及药学上可接受的载体、赋形剂。Another aspect of the present invention provides a pharmaceutical composition comprising a compound represented by formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier and excipient.

本发明另一方面涉及一种式I化合物在制备药物中的用途，其特征在于所述药物可选择性治疗胃癌；优选地，所述药物可选择性抑制胃癌MGC-803细胞株。Another aspect of the present invention relates to the use of a compound of formula I in the preparation of a medicament, characterized in that the medicament can selectively treat gastric cancer; preferably, the medicament can selectively inhibit the gastric cancer MGC-803 cell line.

定义：definition:

“烷基”是指仅仅由碳和氢原子组成，不含有不饱和度，可为C1-6烷基。在一些实施方案中，烷基具有1至6或1至4个碳原子。代表性饱和直链烷基包括但不限于-甲基、-乙基、-正丙基、-正丁基、-正戊基和-正己基；而饱和支链烷基包括但不限于-异丙基、-仲丁基、-异丁基、-叔丁基、-异戊基、2-甲基丁基、3-甲基丁基、2-甲基-戊基、3-甲基戊基、4-甲基戊基、2-甲基己基、3-甲基己基、4-甲基己基、5-甲基己基、2,3-二甲基-丁基等。烷基通过单键连接于母体分子。除非在说明书中另外陈述，否则烷基任选被一个或多个独立地包括以下的取代基取代：酰基、烷基、烯基、炔基、烷氧基、烷基芳基、环烷基。在一非限制性实施方案中，取代的烷基可选自氟甲基、二氟甲基、三氟甲基、2-氟乙基、3-氟丙基、羟基甲基、2-羟基乙基、3-羟基丙基、苯甲基和苯乙基。"Alkyl" means consisting only of carbon and hydrogen atoms, without unsaturation, and may be a C1-6 alkyl group. In some embodiments, the alkyl group has 1 to 6 or 1 to 4 carbon atoms. Representative saturated straight chain alkyl groups include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl; while saturated branched chain alkyl groups include, but are not limited to, -iso Propyl, -sec-butyl, -isobutyl, -tert-butyl, -isoamyl, 2-methylbutyl, 3-methylbutyl, 2-methyl-pentyl, 3-methylpentyl , 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethyl-butyl and the like. The alkyl group is attached to the parent molecule through a single bond. Unless otherwise stated in the specification, alkyl groups are optionally substituted with one or more substituents independently including: acyl, alkyl, alkenyl, alkynyl, alkoxy, alkylaryl, cycloalkyl. In a non-limiting embodiment, the substituted alkyl may be selected from fluoromethyl, difluoromethyl, trifluoromethyl, 2-fluoroethyl, 3-fluoropropyl, hydroxymethyl, 2-hydroxyethyl group, 3-hydroxypropyl, benzyl and phenethyl.

“烷氧基”是指“烷基”通过氧原子与母体分子相连，其中“烷基”具有如上所述的定义。"Alkoxy" means an "alkyl" group attached to the parent molecule through an oxygen atom, wherein "alkyl" has the definition above.

“卤代烷基”是指其中所有氢原子部分或全部被选自氟代基、氯代基、溴代基和碘代基的卤素置换的烷基。在一些实施方案中，所有氢原子都各自被氟代基置换。在一些实施方案中，所有氢原子都各自被氯代基置换。卤代烷基的实例包括-CF3、-CF2CF3、-CF2CF2CF3、-CFCl2、-CF2Cl等。"Haloalkyl" refers to an alkyl group in which all or all of the hydrogen atoms are replaced with halogens selected from the group consisting of fluoro, chloro, bromo, and iodo. In some embodiments, all hydrogen atoms are each replaced with a fluoro group. In some embodiments, all hydrogen atoms are each replaced with a chloro group. Examples of haloalkyl groups include -CF3, -CF2CF3, -CF2CF2CF3, -CFCl2, -CF2Cl, and the like.

在某些实施方案中，药学上可接受的形式是药学上可接受的盐,药学上可接受的盐在本领域中是熟知的。药学上可接受的盐的实例是诸如盐酸、氢溴酸、磷酸、硫酸、高氯酸、乙酸、草酸、顺丁烯二酸、酒石酸、柠檬酸、丁二酸或丙二酸、乙酸、丙酸、乙醇酸、丙酮酸、草酸、乳酸、三氟乙酸、甲烷磺酸、乙烷磺酸、对甲苯磺酸、水杨酸等。In certain embodiments, the pharmaceutically acceptable form is a pharmaceutically acceptable salt, which is well known in the art. Examples of pharmaceutically acceptable salts are such as hydrochloric, hydrobromic, phosphoric, sulfuric, perchloric, acetic, oxalic, maleic, tartaric, citric, succinic or malonic, acetic, propylene Acid, glycolic acid, pyruvic acid, oxalic acid, lactic acid, trifluoroacetic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

“药学上可接受的载体”或“药学上可接受的赋形剂”包括任何和所有溶剂、分散介质、包覆剂、抗细菌剂和抗真菌剂、等张剂和吸收延迟剂等。药学上可接受的载体或赋形剂不破坏公开的化合物的药理学活性，并且在以足以递送治疗量的化合物的剂量施用时是无毒的。药物活性物质的所述介质和试剂的使用在本领域中是熟知的。"Pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like. A pharmaceutically acceptable carrier or excipient does not destroy the pharmacological activity of the disclosed compounds and is non-toxic when administered in doses sufficient to deliver a therapeutic amount of the compound. The use of such media and agents for pharmaceutically active substances is well known in the art.

与现有技术相比，本发明的有益效果是：Compared with the prior art, the beneficial effects of the present invention are:

（1）本发明提供了一类新的具有抗胃癌活性的吲哚酰胺类化合物，拓宽了现有抗胃癌化合物的范围，可作为先导化合物继续优化；(1) The present invention provides a new class of indole amide compounds with anti-gastric cancer activity, which broadens the scope of existing anti-gastric cancer compounds and can be used as lead compounds for further optimization;

（2）本发明化合物具有选择性抑制胃癌细胞的活性，可作为选择性和/或靶向药物，具有良好的开发前景。(2) The compounds of the present invention have the activity of selectively inhibiting gastric cancer cells, can be used as selective and/or targeted drugs, and have good development prospects.

具体实施方式Detailed ways

下面通过实施例来具体说明本发明的内容。在本发明中，以下实施例是为了更好地阐述本发明，并不是用来限制本发明的范围。The content of the present invention will be specifically described by the following examples. In the present invention, the following examples are intended to better illustrate the present invention and are not intended to limit the scope of the present invention.

实施例1Example 1

N-(2-(1H-吲哚-3-基)乙基)-2-羟基-3-甲基苯甲酰胺(化合物R2)N-(2-(1H-Indol-3-yl)ethyl)-2-hydroxy-3-methylbenzamide (Compound R2)

步骤一：在200ml圆底烧瓶加入苯胺（9.3g，0.1mol）和稀盐酸，用30 ml含（7.3g，0.105 mol）亚硝酸钠溶液逐滴加入溶液中。反应1小时抽滤出澄清液体滴加至70ml含（37.8g，0.3 mol）亚硫酸钠溶液中，80℃反应2小时，再滴加约25ml （0.3mol）浓盐酸，95℃反应2小时后置于常温过夜，析出苯肼盐酸盐固体12.6 g。收率：87.1％。Step 1: Add aniline (9.3 g, 0.1 mol) and dilute hydrochloric acid to a 200 ml round-bottomed flask, and add 30 ml (7.3 g, 0.105 mol) of sodium nitrite solution dropwise to the solution. After reacting for 1 hour, the clear liquid was filtered and added dropwise to 70 ml of sodium sulfite solution (37.8 g, 0.3 mol), reacted at 80 °C for 2 hours, and then added dropwise with about 25 ml (0.3 mol) of concentrated hydrochloric acid. After overnight at room temperature, 12.6 g of solid phenylhydrazine hydrochloride was precipitated. Yield: 87.1%.

步骤二：在200ml圆底烧瓶中依次加入步骤一中所得（12.6g，约0.087mol）苯肼盐酸盐，（21.1g，0.10mol）4-氯-1-羟基-丁烷磺酸钠，40ml乙醇，40ml水，再用磷酸氢二钠溶液调节溶液PH至6-7，75℃冷凝回流6小时；用旋转蒸发仪旋去乙醇及少量的水，剩余溶液先用二氯甲烷洗涤，再用碳酸钠溶液调节PH至8-10，再用氯仿洗涤，置于4℃冰箱冷却过夜，析出色胺盐酸盐固体10.3 g。收率：60.3％。Step 2: Add (12.6g, about 0.087mol) phenylhydrazine hydrochloride and (21.1g, 0.10mol) sodium 4-chloro-1-hydroxy-butanesulfonate obtained in step 1 into a 200ml round-bottomed flask in turn, 40ml of ethanol, 40ml of water, adjust the pH of the solution to 6-7 with disodium hydrogen phosphate solution, and condense and reflux at 75°C for 6 hours; use a rotary evaporator to remove ethanol and a small amount of water, and wash the remaining solution with dichloromethane first, then The pH was adjusted to 8-10 with sodium carbonate solution, washed with chloroform, placed in a refrigerator at 4°C to cool overnight, and 10.3 g of trypamine hydrochloride was precipitated as a solid. Yield: 60.3%.

步骤三：在50ml圆底烧瓶中依次加入（0.912g，6 mmol）3-甲基水杨酸，（1.15g，9mmol）EDCI（1-乙基-(3-二甲基氨基丙基)碳酰二亚胺盐酸盐），（1.215g，9 mmol）HOBT（1-羟基苯并三唑），15ml丙酮，常温搅拌2小时；再加入步骤二所得(1.08g，5.5 mmol)色胺盐酸盐，无水碳酸钾，常温搅拌24小时；柱色谱纯化，洗脱剂为：乙酸乙酯 ：石油醚=1 ：2。得1.31g化合物R2（收率：81.2％），其为白色固体，熔点为：165-167℃。¹H NMR (400 MHz,CDCl₃) δ 12.39 (s, 1H), 8.22 (s, 1H), 7.61 (s, 1H), 7.33 (d,J = 8.5 Hz, 1H),7.29 (s, 1H), 7.20 (d,J = 7.9 Hz, 1H), 7.11 (s, 1H), 7.08 (d,J = 7.8 Hz,1H), 6.82 (s, 1H), 6.62 (d,J = 7.3 Hz, 1H), 6.37 (s, 1H), 3.77 (d,J = 5.1Hz, 2H), 3.07 (d,J = 5.9 Hz, 2H), 2.33 (s, 3H)。Step 3: Add (0.912g, 6 mmol) 3-methylsalicylic acid, (1.15g, 9 mmol) EDCI (1-ethyl-(3-dimethylaminopropyl) carbon to a 50ml round-bottomed flask in turn Diimide hydrochloride), (1.215g, 9 mmol) HOBT (1-hydroxybenzotriazole), 15ml acetone, stir at room temperature for 2 hours; then add the (1.08g, 5.5 mmol) tryptamine salt obtained in step two acid salt, anhydrous potassium carbonate, stirring at room temperature for 24 hours; column chromatography purification, eluent: ethyl acetate: petroleum ether=1:2. 1.31 g of compound R2 was obtained (yield: 81.2%) as a white solid, melting point: 165-167°C.¹ H NMR (400 MHz, CDCl₃ ) δ 12.39 (s, 1H), 8.22 (s, 1H), 7.61 (s, 1H), 7.33 (d,J = 8.5 Hz, 1H), 7.29 (s, 1H) , 7.20 (d,J = 7.9 Hz, 1H), 7.11 (s, 1H), 7.08 (d,J = 7.8 Hz, 1H), 6.82 (s, 1H), 6.62 (d,J = 7.3 Hz, 1H) , 6.37 (s, 1H), 3.77 (d,J = 5.1 Hz, 2H), 3.07 (d,J = 5.9 Hz, 2H), 2.33 (s, 3H).

实施例2-5Example 2-5

参照实施例1的方法，制备得到化合物R3-R6，相应的数据如下所示：With reference to the method of Example 1, compounds R3-R6 are prepared, and the corresponding data are as follows:

化合物compoundR（n=1）R (n=1)熔点/℃Melting point/℃¹H NMR¹H NMRR3R34-CH34-CH3165-167165-167¹H NMR (400 MHz, CDCl₃) δ 12.72 (s, 1H), 8.02(s, 1H), 7.43 (s, 1H), 7.31 (d, <i>J</i> = 8.3Hz, 1H), 7.29 (s, 1H), 7.25 (d, <i>J</i> = 7.2Hz, 1H), 7.10 (s, 1H), 7.07 (s, 1H), 6.97(d, <i>J</i> = 7.9 Hz, 1H), 6.67 (t, <i>J</i> = 7.6 Hz,1H), 6.42 (s, 1H), 3.83 – 3.73 (m, 2H), 3.10(t, <i>J</i> = 6.3 Hz, 2H), 2.46 (s, 3H)¹H NMR (400 MHz, CDCl₃) δ 12.72 (s, 1H), 8.02(s, 1H), 7.43 (s, 1H), 7.31 (d, <i>J</i> = 8.3Hz, 1H), 7.29 (s, 1H), 7.25 (d, <i>J</i> = 7.2Hz, 1H), 7.10 (s, 1H), 7.07 ( s, 1H), 6.97(d, <i>J</i> = 7.9 Hz, 1H), 6.67 (t, <i>J</i> = 7.6 Hz, 1H), 6.42 (s, 1H), 3.83 – 3.73 (m, 2H), 3.10(t, <i>J</i> = 6.3 Hz, 2H), 2.46 (s, 3H)R4R44-Cl4-Cl158-160158-160¹H NMR (400 MHz, CDCl₃) δ 12.55 (s, 1H), 8.12(s, 1H), 7.51 (s, 1H), 7.22 (d, <i>J</i> = 8.6Hz, 1H), 7.18 (s, 1H), 7.16 (d, <i>J</i> = 7.3Hz, 1H), 7.09 (d, <i>J</i> = 8.6 Hz, 1H), 7.01 (s,1H), 6.94 (d, <i>J</i> = 7.9 Hz, 1H), 6.60 (t, <i>J</i>= 7.6 Hz, 1H), 6.32 (s, 1H), 3.67 (q, <i>J</i> =6.0 Hz, 2H), 2.96 (t, <i>J</i> = 6.5 Hz, 2H)¹H NMR (400 MHz, CDCl₃) δ 12.55 (s, 1H), 8.12(s, 1H), 7.51 (s, 1H), 7.22 (d, <i>J</i> = 8.6Hz, 1H), 7.18 (s, 1H), 7.16 (d, <i>J</i> = 7.3Hz, 1H), 7.09 (d, <i>J< /i> = 8.6 Hz, 1H), 7.01 (s, 1H), 6.94 (d, <i>J</i> = 7.9 Hz, 1H), 6.60 (t, <i>J</i>= 7.6 Hz, 1H), 6.32 (s, 1H), 3.67 (q, <i>J</i> =6.0 Hz, 2H), 2.96 (t, <i>J</i> = 6.5 Hz, 2H)R5R55-Cl5-Cl170-173170-173¹H NMR (400 MHz, CDCl₃) δ 12.35 (s, 1H), 8.20(s, 1H), 7.60 (s, 1H), 7.46 (d, <i>J</i> = 8.8Hz, 1H), 7.33 (d, <i>J</i> = 8.6 Hz, 1H), 7.28(s, 2H), 7.21 (d, <i>J</i> = 8.6 Hz, 1H), 7.12 (s,1H), 6.90 (d, <i>J</i> = 8.8 Hz, 1H), 6.38 (s,1H), 3.77 (q, <i>J</i> = 6.4 Hz, 2H), 3.07 (t, <i>J</i>= 6.6 Hz, 2H)¹H NMR (400 MHz, CDCl₃) δ 12.35 (s, 1H), 8.20(s, 1H), 7.60 (s, 1H), 7.46 (d, <i>J</i> = 8.8Hz, 1H), 7.33 (d, <i>J</i> = 8.6 Hz, 1H), 7.28(s, 2H), 7.21 (d, <i>J< /i> = 8.6 Hz, 1H), 7.12 (s, 1H), 6.90 (d, <i>J</i> = 8.8 Hz, 1H), 6.38 (s, 1H), 3.77 (q, <i> J</i> = 6.4 Hz, 2H), 3.07 (t, <i>J</i>= 6.6 Hz, 2H)R6R65-Br5-Br174-177174-177¹H NMR (400 MHz, CDCl₃) δ 12.40 (s, 1H), 8.15(s, 1H), 7.67 (d, <i>J</i> = 7.8 Hz, 1H), 7.46(s, 1H), 7.44 (s, 1H), 7.27 (s, 1H), 7.23(s, 1H), 7.20 (d, <i>J</i> = 7.4 Hz, 1H), 7.12 (s,1H), 6.90 (d, <i>J</i> = 8.8 Hz, 1H), 6.36 (s,1H), 3.92 – 3.68 (m, 2H), 3.13 (t, <i>J</i> = 6.5Hz, 2H)¹H NMR (400 MHz, CDCl₃) δ 12.40 (s, 1H), 8.15(s, 1H), 7.67 (d, <i>J</i > = 7.8 Hz, 1H), 7.46(s, 1H), 7.44 (s, 1H), 7.27 (s, 1H), 7.23(s, 1H), 7.20 (d, <i>J</i> = 7.4 Hz, 1H), 7.12 (s, 1H), 6.90 (d, <i>J</i> = 8.8 Hz, 1H), 6.36 (s, 1H), 3.92 – 3.68 (m, 2H), 3.13 (t , <i>J</i> = 6.5Hz, 2H)

实施例6 体外活性测试Example 6 In vitro activity test

细胞株选用MGC-803(人胃癌细胞)、HepG2(人肝癌细胞)，MCF-7（人乳腺癌细胞），A549（人肺癌细胞）和HELA（人宫颈癌细胞）。The cell lines selected were MGC-803 (human gastric cancer cells), HepG2 (human liver cancer cells), MCF-7 (human breast cancer cells), A549 (human lung cancer cells) and HELA (human cervical cancer cells).

培养液为DMEM+15％ NBS+ 双抗The culture medium is DMEM + 15% NBS + double antibody

样品液配制：用DMSO(Merck) 溶解后，浓度为33333.33μmol/LPreparation of sample solution: after dissolving with DMSO (Merck), the concentration is 33333.33μmol/L

1.中板 将细胞悬液加入96孔板，每孔100μl（每孔的细胞含量约为8000个/孔），置于37℃、5％ CO2培养箱培育24h；1. Medium plate: Add the cell suspension to a 96-well plate, 100 μl per well (the cell content of each well is about 8000 cells/well), and incubate it in a 37°C, 5% CO2 incubator for 24 hours;

2.加药 用培养基配制样品液梯度浓度（256，128，64，32，16，8，4，2 μmol/L），再弃去96孔板上原有培养基，加入药物不同浓度的培养基，每孔100μL，每个浓度做5个副孔。其余孔用含有3‰的DMSO的培养基做对照，置37℃、5％培养箱培育48h；2. Add the medicinal medium to prepare the gradient concentration of the sample solution (256, 128, 64, 32, 16, 8, 4, 2 μmol/L), then discard the original medium on the 96-well plate, and add different concentrations of the drug to culture Base, 100 μL per well, 5 auxiliary wells for each concentration. The rest of the wells were controlled with a medium containing 3‰ of DMSO, and incubated at 37°C in a 5% incubator for 48h;

3.MTT法测试 取出96孔板，避光条件下每孔加入10μLMTT(3-(4，5- 二甲基噻唑-2- 基)-2，5- 二苯基四唑翁溴化物，5mg/mL）溶液，再放入培养箱内放置4h；4小时后弃去培养液，每孔加入150μLDMSO溶解震荡，用MK-2 全自动酶标仪测490nm OD 值，计算半数抑制浓度IC50。3. MTT method test Take out the 96-well plate, and add 10 μL MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, 5mg to each well under dark conditions) /mL) solution, and then placed in the incubator for 4 hours; after 4 hours, the culture medium was discarded, 150 μL DMSO was added to each well to dissolve and shake, and the OD value at 490 nm was measured with an MK-2 automatic microplate reader, and the median inhibitory concentration IC50 was calculated.

测试结果如下：The test results are as follows:

由上表可以看，本发明的化合物能够选择性抑制胃癌细胞，尤其是选择性抑制MGC-803(人胃癌细胞)细胞株。因而，可以用做选择性治疗胃癌的药物，具有良好的开发前景。It can be seen from the above table that the compounds of the present invention can selectively inhibit gastric cancer cells, especially the MGC-803 (human gastric cancer cell) cell line. Therefore, it can be used as a drug for selective treatment of gastric cancer, and has a good development prospect.

Claims (10)

1. A compound of formula I, or a pharmaceutically acceptable salt thereof, having the structure:

wherein: each R is independently selected from halogen, C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl;

n is selected from 1, 2,3 or 4.

2. A compound of formula I according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R is each independently selected from fluoro, chloro, bromo, methyl, ethyl, methoxy, ethoxy, trifluoromethyl; n is selected from 1 or 2.

3. A compound of formula I according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, wherein each R is independently selected from methyl, chloro, bromo; n is 1.

4. A compound of formula I according to claim 1, selected from the following compounds:

5. a process for the preparation of a compound of formula I according to claim 1, which reaction scheme is as follows:

wherein R and n are as defined in claim 1;

the specific reaction steps are as follows:

the method comprises the following steps: aniline and hydrochloric acid are added to a reaction vessel, and a sodium nitrite solution is added dropwise to the solution. Carrying out suction filtration after reacting for 1 hour, dropwise adding the filtrate into a sodium sulfite solution, reacting for 2 hours at 80 ℃, dropwise adding concentrated hydrochloric acid, reacting for 2 hours at 95 ℃, standing at normal temperature overnight, and separating out phenylhydrazine hydrochloride solid;

step two: sequentially adding phenylhydrazine hydrochloride, 4-chloro-1-hydroxy-butane sodium sulfonate, ethanol and water into a reaction vessel, adjusting the pH value of the solution to 6-7, and reacting for 6 hours at 75 ℃; distilling under reduced pressure to remove ethanol and part of water, washing the residual solution with dichloromethane, adjusting the pH value to 8-10, washing with chloroform, and cooling in a refrigerator at 4 ℃ overnight to separate out tryptamine hydrochloride solid;

step three: sequentially adding salicylic acid derivative, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, 1-hydroxybenzotriazole and acetone into a reaction vessel, and stirring at normal temperature for 2 hours; adding the tryptamine hydrochloride obtained in the step two and anhydrous potassium carbonate, and stirring at normal temperature until the reaction is complete; purifying by column chromatography to obtain the compound shown in formula I.

6. The method of claim 5, wherein:

the molar ratio of the aniline to the sodium nitrite to the sodium sulfite in the first step is 1:1-1.5: 2-4.5;

the molar ratio of the phenylhydrazine hydrochloride in the step II to the sodium 4-chloro-1-hydroxy-butane sulfonate is 1: 1-1.5;

and step three, the molar ratio of the salicylic acid derivative, the 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, the 1-hydroxybenzotriazole and the tryptamine hydrochloride is 1:1-2:1-2: 0.5-1.5.

7. The method of claim 6, wherein:

the molar ratio of the aniline, the sodium nitrite and the sodium sulfite in the first step is 1:1.05: 3;

the molar ratio of the phenylhydrazine hydrochloride in the step II to the sodium 4-chloro-1-hydroxy-butane sulfonate is 1: 1-1.2;

and step three, the molar ratio of the salicylic acid derivative, the 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride, the 1-hydroxybenzotriazole and the tryptamine hydrochloride is 1:1-1.5:1-1.5: 0.8-1.2.

8. A pharmaceutical composition comprising a compound of formula I as shown in any one of claims 1 to 4 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

9. Use of a compound according to any one of claims 1 to 4 or a pharmaceutical composition according to claim 8 for the manufacture of a medicament for the treatment of gastric cancer.

10. The use of claim 9, wherein the medicament selectively inhibits gastric cancer MGC-803 cell line.