技术领域technical field
本发明涉及化学药物领域，具体地说是以色氨酸甲酯为起始物，合成一系列的吲哚并[3,2-a]咔唑衍生物和衍生物盐及其在制备抗肿瘤药物中的应用。The invention relates to the field of chemical medicines, specifically using tryptophan methyl ester as a starting material to synthesize a series of indolo[3,2-a]carbazole derivatives and derivative salts and their use in the preparation of antitumor application in medicine.
背景技术Background technique
肿瘤一直都是人类生命的一大威胁，而人类与肿瘤的斗争一直在继续，其中最为有效的方法就是药物治疗，而在治疗肿瘤的很多药物中存在低效高毒的现象，所以开发和研究具有抗肿瘤活性且毒性较低的药物，一直是现在医药领域的一个重要方向，相关领域的研究者们一直致力于找到一类有效低毒的化合物，以之得以解决人类的这一大难题。Tumors have always been a major threat to human life, and the struggle between humans and tumors has continued. The most effective method is drug therapy, and many drugs for treating tumors have low efficiency and high toxicity, so development and research Drugs with anti-tumor activity and low toxicity have always been an important direction in the field of medicine. Researchers in related fields have been working hard to find a class of effective and low-toxic compounds to solve this big problem for humans.
吲哚并[3,2-a]咔唑类生物碱是较为新颖的天然产物，直到2002年才从海洋生物海绵中被发现，具有广泛的生物活性特别是抗肿瘤活性，一直备受较为广泛的关注，但由于合成方法较少及天然来源有限，迄今为止围绕此化合物的药化和活性研究很少。本发明人近期发展了一条对于此类咔唑生物碱的合成方法，可以较为方便的得到不同基团取代的吲哚并[3,2-a]咔唑衍生物。在之前的研究中，对吲哚并[3,2-a]咔唑类生物碱的抗生物活性研究很少，尤其是对其抗肿瘤活性的报道相对较少，对抗肿瘤的种类也相对较少，为了拓展此类化合物的化学空间和生物学空间，利用吲哚并[3,2-a]咔唑这一较为新颖的天然产物母核作为分子模板，通过引入药效团来设计和合成一系列的化合物，希望在保持活性和减少毒性的基础上，合成具有较好理化性质和代谢稳定性的药物作为先导化合物，并且研究其对多种肿瘤细胞的生物活性，为抗肿瘤新药的开发提供物质基础。Indolo[3,2-a]carbazole alkaloids are relatively new natural products, which were not discovered from marine sponges until 2002. They have a wide range of biological activities, especially anti-tumor activities, and have been widely studied. However, due to the lack of synthetic methods and limited natural sources, so far there are few studies on the pharmacology and activity of this compound. The present inventor has recently developed a synthesis method for this kind of carbazole alkaloid, which can conveniently obtain indolo[3,2-a]carbazole derivatives substituted by different groups. In previous studies, there were few studies on the antibiological activity of indolo[3,2-a]carbazole alkaloids, especially the reports on their antitumor activity were relatively few, and the antitumor types were relatively few. In order to expand the chemical space and biological space of such compounds, indolo[3,2-a]carbazole, a relatively new natural product core, was used as a molecular template to design and synthesize by introducing pharmacophore A series of compounds, hoping to synthesize drugs with better physical and chemical properties and metabolic stability as lead compounds on the basis of maintaining activity and reducing toxicity, and to study their biological activities on various tumor cells, for the development of new anti-tumor drugs Provide a material basis.
发明内容Contents of the invention
本发明的目的是利用吲哚并[3,2-a]咔唑这一较为新颖的天然产物母核作为分子模板，通过引入药效团来设计和合成一系列的化合物，制备新的、结构多样的吲哚并[3,2-a]咔唑衍生物及其药学上可接受的盐，研究具有抗各种肿瘤活性，为抗肿瘤新药的制备提供物质基础。The purpose of the present invention is to use indolo[3,2-a]carbazole, a relatively novel natural product core, as a molecular template, to design and synthesize a series of compounds by introducing pharmacophore, to prepare new, structural A variety of indolo[3,2-a]carbazole derivatives and their pharmaceutically acceptable salts have been studied to have anti-tumor activities, providing a material basis for the preparation of new anti-tumor drugs.
本发明一种吲哚并[3,2-a]咔唑衍生物，其特征在于，其结构通式为式（１）：An indolo[3,2-a]carbazole derivative of the present invention is characterized in that its general structural formula is formula (1):
式（１）中：In formula (1):
R1为氢、羟基、；R1 is hydrogen, hydroxyl, ;
R2为氢、卤原子、羟基、C1～C4烷氧基、C1～C4烷酰基；R2 ishydrogen , a halogen atom, a hydroxyl group, a C1-C4 alkoxy group, a C1-C4 alkanoyl group;
R3为由通式（2）所示的取代基团：R3 is a substituent group represented by general formula (2):
式（2）中，n=1～6，R选自烷氨基、酰胺基、杂环烷基、芳基、取代苯基、芳杂环基；所指吲哚并[3,2-a]咔唑衍生物简称衍生物还包括其生成的盐类；所指的衍生物或衍生物盐具有抑制肿瘤的活性。In formula (2), n=1~6, R is selected from alkylamino group, amido group, heterocycloalkyl group, aryl group, substituted phenyl group, aromatic heterocyclic group; refers to indolo[3,2-a] The carbazole derivatives referred to as derivatives also include their salts; the referred derivatives or derivative salts have the activity of inhibiting tumors.
4.以上所指的一种吲哚并[3,2-a]咔唑衍生物衍生物盐为药学上可接受的盐酸盐、硫酸盐、甲磺酸盐、对甲苯磺酸盐、马来酸盐、富马酸盐、酒石酸盐；具体来说上述通式（１）中R1代表R1代表氢、羟基、；4. A derivative salt of an indolo[3,2-a]carbazole derivative referred to above is pharmaceutically acceptable hydrochloride, sulfate, methanesulfonate, p-toluenesulfonate, horseradish Forate, fumarate, tartrate; specifically, in the above general formula (1), R1 represents R1 represents hydrogen, hydroxyl, ;
R2代表氢、甲氧基、氯或溴；R3代表，而衍生物盐为药学上可接受的盐酸盐，衍生物或衍生物盐具有抗肿瘤活性；合成的部分化合物或其盐如表1，其中优选已合成的化合物C1、C3、C11、C12、C13、C14、C15、C16、C27、C28或其盐酸盐具有显著的抗肿瘤活性；这一类吲哚并[3,2-a]咔唑衍生物的应用，是用于制备抗人红白血病细胞HEL，骨髓性白血病细胞K562-1，人黑素瘤细胞WM9-1，乳腺癌细胞MDA231，B淋巴母细胞TK-6，FV-P诱导红白血病(小鼠细胞)DP17-17，前列腺癌细胞PC-3这些细胞形成的肿瘤的药物或药剂。R2 represents hydrogen, methoxy, chlorine or bromine; R3 represents , and derivative salts are pharmaceutically acceptable hydrochlorides, and derivatives or derivative salts have antitumor activity; some compounds or their salts synthesized are shown in Table 1, among which the synthesized compounds C1, C3, C11, and C12 are preferred , C13, C14, C15, C16, C27, C28 or their hydrochlorides have significant antitumor activity; the application of this kind of indolo[3,2-a]carbazole derivatives is used to prepare anti-human Erythroleukemia cells HEL, myeloid leukemia cells K562-1, human melanoma cells WM9-1, breast cancer cells MDA231, B lymphoblastoid cells TK-6, FV-P induced erythroleukemia (mouse cells) DP17-17, Prostate cancer cells PC-3 Drugs or agents for tumors formed by these cells.
表1：合成的部分化合物Table 1: Some compounds synthesized
表2表1化合物的核磁数据表征Table 2 NMR data characterization of the compounds in Table 1
优选C1、C3、C11、C12、C13、C14、C15、C16、C27、C28，对这十种优选的吲哚并[3,2-a]咔唑衍生物将进行实施例说明。Preferred are C1, C3, C11, C12, C13, C14, C15, C16, C27, and C28, and these ten preferred indolo[3,2-a]carbazole derivatives will be described in examples.
对化合物进行体外和体内的抗肿瘤试验，表明其具有显著的抑制肿瘤的活性，因此本发明化合物可以用于制备抗肿瘤的药物或药剂。Anti-tumor tests in vitro and in vivo on the compound show that it has significant tumor-inhibiting activity, so the compound of the present invention can be used to prepare anti-tumor drugs or medicaments.
发明效果：在本发明已合成的化合物中，优选出C1、C3、C11、C12、C13、C14、C15、C16、C27、C28或其盐为代表，对人红白血病细胞HEL，骨髓性白血病细胞K562-1，人黑素瘤细胞WM9-1，乳腺癌细胞MDA231，B淋巴母细胞TK-6，FV-P诱导红白血病(小鼠细胞)DP17-17，前列腺癌细胞PC-3多种肿瘤细胞进行药物实验，发现均具有显著的抗肿瘤作用。Effect of the invention: Among the compounds synthesized in the present invention, C1, C3, C11, C12, C13, C14, C15, C16, C27, C28 or their salts are preferred, which are effective against human erythroleukemia cells HEL and myeloid leukemia cells K562-1, human melanoma cell WM9-1, breast cancer cell MDA231, B lymphoblastoid cell TK-6, FV-P induced erythroleukemia (mouse cell) DP17-17, prostate cancer cell PC-3 various tumors Drug experiments were carried out on the cells, and it was found that they all had significant anti-tumor effects.
说明书附图Instructions attached
图1为原料F的合成路线；Fig. 1 is the synthetic route of raw material F;
图2为原料M的合成路线；Fig. 2 is the synthetic route of raw material M;
图3为原料P的合成路线；Fig. 3 is the synthetic route of raw material P;
图4为原料U的合成路线；Fig. 4 is the synthetic route of raw material U;
图5为实施例12中C1对K562细胞生长的影响；从中可以看出化合物C1的浓度与对K562细胞生长的抑制率呈正相关，对K562细胞有较为明显的抑制作用；Figure 5 is the effect of C1 on the growth of K562 cells in Example 12; it can be seen that the concentration of compound C1 is positively correlated with the inhibition rate of K562 cell growth, and has a more obvious inhibitory effect on K562 cells;
图6为实施例13中C1在72h时对K562细胞细胞周期的影响；从中可以看出C1对K562细胞细胞周期有明显的影响，阻断在G2期；Figure 6 is the effect of C1 on the cell cycle of K562 cells at 72h in Example 13; it can be seen that C1 has a significant effect on the cell cycle of K562 cells, blocking in the G2 phase;
图7为实施例14中C1（1mg/kg)给药两周对感染F-MuLV小鼠脾脏重量的影响；从中可以看出患病的小鼠的脾脏基本恢复正常，也就说化合物C1对感染F-MuLV的小鼠有较为明显是治疗作用；Fig. 7 is the impact of two weeks of administration of C1 (1mg/kg) on the spleen weight of infected F-MuLV mice in Example 14; It can be seen from this that the spleen of the sick mice returns to normal basically, that is to say that compound C1 has the effect on the spleen weight of F-MuLV mice Mice infected with F-MuLV have a more obvious therapeutic effect;
图8为实施例14中C1（1mg/kg)给药两周对感染F-MuLV小鼠红细胞比容的影响，从中可以看出，病的小鼠的红细胞比容基本恢复正常，说明化合物C1对感染F-MuLV的小鼠有较为明显是治疗作用。Fig. 8 is the influence of two weeks of administration of C1 (1mg/kg) on the hematocrit of infected F-MuLV mice in Example 14, from which it can be seen that the hematocrit of the sick mice returns to normal basically, indicating that compound C1 It has a more obvious therapeutic effect on mice infected with F-MuLV.
具体实施方式detailed description
下面结合具体实例对本发明做详细阐述，需要说明的是，下述实施例仅是用于说明，而并非用于限制本发明。本领域技术人员根据本发明的教导所做出的各种变化均应在本申请权利要求所要求的保护范围之内。The present invention will be described in detail below in conjunction with specific examples. It should be noted that the following examples are only for illustration, not for limiting the present invention. Various changes made by those skilled in the art according to the teaching of the present invention shall be within the scope of protection required by the claims of the present application.
原料F的制备Preparation of raw material F
图1为制备化合物F的技术路线。具体方法：称取化合物A（4.03mmol）溶于10mL超干四氢呋喃中，加入三乙胺（4.8mmol，1.2eq），然后于-78℃下加入次氯酸叔丁基酯（4.83mmol,1.2eq），反应40min后，依次加入化合物B（8.06mmol,2.0eq）和三氟化硼.乙醚（16.09mmol,4.0eq），升至室温反应5h，缓慢加入饱和的碳酸氢钠水溶液调pH=7，乙酸乙酯萃取（3×10mL），合并有机层，无水硫酸镁干燥，过滤，减压浓缩得粗产品，再经柱层析分离纯化得化合物C；将化合物C（0.97mmol）溶于甲醇/二氯甲烷（v/v=1:1）的混合溶剂中，加入80％的水合肼（3.88mmol,4.0eq），室温反应过夜，过滤，取滤液减压浓缩得粗产品，再经柱层析分离纯化得到化合物D；化合物D（0.70mmol）溶于乙腈/醋酸缓冲液（4mL,v/v=1:1）的混合溶剂中，加入七水合硫酸锌（0.35mmol）和乙醛酸钠（7.0mmol），室温反应0.5h，然后加入10mL水淬灭反应，乙酸乙酯萃取（3×10mL），合并有机层，无水Na2SO4干燥、过滤、减压浓缩得粗产物，将此粗产物溶于3mL的1,4-二氧六环中，然后加入0.1mL三氟乙酸，加热回流反应5-8h，冷却后，减压浓缩，经柱层析分离纯化得到目化合物E；称取化合物E（0.38mmol）溶于MeOH/H2O/DMSO（2mL,v/v/v=2:1:1）的混合溶剂中，加入3.8mmolNaOH，室温反应26-30h（TLC监测反应完全），然后加入3mL5%的盐酸溶液，乙酸乙酯萃取，合并有机相，无水Na2SO4干燥、过滤，取滤液减压浓缩化合物F。Figure 1 is the technical route for preparing compound F. Specific method: weigh compound A (4.03mmol) and dissolve it in 10mL ultra-dry tetrahydrofuran, add triethylamine (4.8mmol, 1.2eq), then add tert-butyl hypochlorite (4.83mmol, 1.2 eq), after reacting for 40 minutes, sequentially add compound B (8.06mmol, 2.0eq) and boron trifluoride. Ether (16.09mmol, 4.0eq), rise to room temperature and react for 5h, slowly add saturated aqueous sodium bicarbonate solution to adjust the pH= 7. Extract with ethyl acetate (3×10mL), combine the organic layers, dry over anhydrous magnesium sulfate, filter, concentrate under reduced pressure to obtain a crude product, and then separate and purify by column chromatography to obtain compound C; dissolve compound C (0.97mmol) In the mixed solvent of methanol/dichloromethane (v/v=1:1), add 80% hydrazine hydrate (3.88mmol, 4.0eq), react at room temperature overnight, filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Compound D was obtained by separation and purification by column chromatography; compound D (0.70 mmol) was dissolved in a mixed solvent of acetonitrile/acetic acid buffer (4 mL, v/v=1:1), and zinc sulfate heptahydrate (0.35 mmol) and acetonitrile were added Sodium alkydrate (7.0mmol), reacted at room temperature for 0.5h, then added 10mL of water to quench the reaction, extracted with ethyl acetate (3×10mL), combined the organic layers, dried over anhydrous Na2 SO4 , filtered, and concentrated under reduced pressure to obtain crude Product, this crude product was dissolved in 3mL of 1,4-dioxane, then added 0.1mL of trifluoroacetic acid, heated to reflux for 5-8h, cooled, concentrated under reduced pressure, separated and purified by column chromatography to obtain the target Compound E; weigh compound E (0.38mmol) and dissolve it in a mixed solvent of MeOH/H2 O/DMSO (2mL, v/v/v=2:1:1), add 3.8mmol NaOH, and react at room temperature for 26-30h ( TLC monitors that the reaction is complete), then 3 mL of 5% hydrochloric acid solution was added, extracted with ethyl acetate, the organic phases were combined, dried over anhydrous Na2 SO4 , filtered, and the filtrate was taken to concentrate compound F under reduced pressure.
实施例1Example 1
C1的制备：Preparation of C1:
制备C1的技术路线为：The technical route for preparing C1 is:
具体操作方法为：将0.2mmol化合物F溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物G，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到白色固体59.3mg，产率为80%;The specific operation method is: dissolve 0.2 mmol of compound F in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound G, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 59.3 mg of white solid with a yield of 80%;
C1的NMR数据见表2。See Table 2 for the NMR data of C1.
实施例2Example 2
C3的制备：Preparation of C3:
制备C3的技术路线为：The technical route for preparing C3 is:
具体操作方法为：将0.2mmol化合物F溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物H，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到黄色固体57.8mg，产率为73%;The specific operation method is as follows: dissolve 0.2 mmol of compound F in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound H, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 57.8 mg of a yellow solid with a yield of 73%;
C3的NMR数据见表2。See Table 2 for the NMR data of C3.
原料M的制备Preparation of raw material M
图2为原料M的制备技术路线具体操作方法：称取化合物I（4.03mmol）溶于10mL超干四氢呋喃中，加入三乙胺（4.8mmol，1.2eq），然后于-78℃下加入次氯酸叔丁基酯（4.83mmol,1.2eq），反应40min后，依次加入化合物B（8.06mmol,2.0eq）和三氟化硼.乙醚（16.09mmol,4.0eq），升至室温反应5h，缓慢加入饱和的碳酸氢钠水溶液调pH=7，乙酸乙酯萃取（3×10mL），合并有机层，无水硫酸镁干燥，过滤，减压浓缩得粗产品，再经柱层析分离纯化得化合物J；将化合物J（0.97mmol）溶于甲醇/二氯甲烷（v/v=1:1）的混合溶剂中，加入80％的水合肼（3.88mmol,4.0eq），室温反应过夜，过滤，取滤液减压浓缩得粗产品，再经柱层析分离纯化得到化合物K；化合物K（0.70mmol）溶于乙腈/醋酸缓冲液（4mL,v/v=1:1）的混合溶剂中，加入七水合硫酸锌（0.35mmol）和乙醛酸钠（7.0mmol），室温反应0.5h，然后加入10mL水淬灭反应，乙酸乙酯萃取（3×10mL），合并有机层，无水Na2SO4干燥、过滤、减压浓缩得粗产物，将此粗产物溶于3mL的1,4-二氧六环中，然后加入0.1mL三氟乙酸，加热回流反应5-8h，冷却后，减压浓缩，经柱层析分离纯化得到目化合物L；称取化合物L（0.38mmol）溶于MeOH/H2O/DMSO（2mL,v/v/v=2:1:1）的混合溶剂中，加入3.8mmolNaOH，室温反应26-30h（TLC监测反应完全），然后加入3mL5%的盐酸溶液，乙酸乙酯萃取，合并有机相，无水Na2SO4干燥、过滤，取滤液减压浓缩化合物M。Figure 2 shows the technical route for the preparation of raw material M. The specific operation method: Weigh compound I (4.03mmol) and dissolve it in 10mL ultra-dry tetrahydrofuran, add triethylamine (4.8mmol, 1.2eq), and then add hypochlorite at -78°C Acetate tert-butyl ester (4.83mmol, 1.2eq), after reacting for 40min, add compound B (8.06mmol, 2.0eq) and boron trifluoride. Ether (16.09mmol, 4.0eq) successively, rise to room temperature and react for 5h, slowly Add saturated aqueous sodium bicarbonate solution to adjust the pH=7, extract with ethyl acetate (3×10mL), combine the organic layers, dry over anhydrous magnesium sulfate, filter, concentrate under reduced pressure to obtain the crude product, and then separate and purify the compound by column chromatography J; Dissolve compound J (0.97mmol) in a mixed solvent of methanol/dichloromethane (v/v=1:1), add 80% hydrazine hydrate (3.88mmol, 4.0eq), react overnight at room temperature, filter, The filtrate was concentrated under reduced pressure to obtain a crude product, which was separated and purified by column chromatography to obtain compound K; compound K (0.70 mmol) was dissolved in a mixed solvent of acetonitrile/acetic acid buffer (4 mL, v/v=1:1), and added Zinc sulfate heptahydrate (0.35mmol) and sodium glyoxylate (7.0mmol), react at room temperature for 0.5h, then add 10mL of water to quench the reaction, extract with ethyl acetate (3×10mL), combine the organic layers, anhydrousNa2SO4. Dry, filter, and concentrate under reduced pressure to obtain a crude product. Dissolve the crude product in 3 mL of 1,4-dioxane, then add 0.1 mL of trifluoroacetic acid, and heat under reflux for 5-8 hours. After cooling, depressurize Concentrate, separate and purify by column chromatography to obtain the target compound L; weigh compound L (0.38mmol) and dissolve it in a mixed solvent of MeOH/H2 O/DMSO (2mL, v/v/v=2:1:1), Add 3.8mmol NaOH, react at room temperature for 26-30h (TLC monitors the reaction is complete), then add 3mL of 5% hydrochloric acid solution, extract with ethyl acetate, combine the organic phases, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate compound M under reduced pressure .
实施例3Example 3
C11的制备Preparation of C11
制备C11的技术路线为：The technical route for preparing C11 is:
具体操作方法为：将0.2mmol化合物M溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物G，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到白色固体50.2mg，产率为65%;The specific operation method is: dissolve 0.2 mmol of compound M in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound G, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 50.2 mg of a white solid with a yield of 65%;
C11的NMR数据见表2。See Table 2 for the NMR data of C11.
实施例4Example 4
C12的制备Preparation of C12
制备C12的技术路线为：The technical route for preparing C12 is:
具体操作方法为：将0.2mmol化合物M溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物H，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到黄色固体58.6mg，产率为71%;The specific operation method is: dissolve 0.2 mmol of compound M in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound H, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 58.6 mg of a yellow solid with a yield of 71%;
C12的NMR数据见表2。The NMR data of C12 are shown in Table 2.
原料P的制备Preparation of raw material P
图3为制备化合物P的技术路线。具体操作方法：称取化合物I（4.03mmol）溶于10mL超干四氢呋喃中，加入三乙胺（4.8mmol，1.2eq），然后于-78℃下加入次氯酸叔丁基酯（4.83mmol,1.2eq），反应40min后，依次加入化合物B（8.06mmol,2.0eq）和三氟化硼.乙醚（16.09mmol,4.0eq），升至室温反应5h，缓慢加入饱和的碳酸氢钠水溶液调pH=7，乙酸乙酯萃取（3×10mL），合并有机层，无水硫酸镁干燥，过滤，减压浓缩得粗产品，再经柱层析分离纯化得化合物J；将化合物J（0.97mmol）溶于甲醇/二氯甲烷（v/v=1:1）的混合溶剂中，加入80％的水合肼（3.88mmol,4.0eq），室温反应过夜，过滤，取滤液减压浓缩得粗产品，再经柱层析分离纯化得到化合物K；化合物K（0.70mmol）溶于乙腈/醋酸缓冲液（4mL,v/v=1:1）的混合溶剂中，加入七水合硫酸锌（0.35mmol）和乙醛酸钠（7.0mmol），室温反应0.5h，然后加入10mL水淬灭反应，乙酸乙酯萃取（3×10mL），合并有机层，无水Na2SO4干燥、过滤、减压浓缩得粗产物，将此粗产物溶于3mL的1,4-二氧六环中，然后加入0.1mL三氟乙酸，加热回流反应5-8h，冷却后，减压浓缩，经柱层析分离纯化得到目化合物L；将化合物L和化合物N溶于3mL丙酮，加入碳酸钾（4.0eq）、四丁基碘化铵（1.2eq），加热回流5h，减压蒸去丙酮，加入10mL水，乙酸乙酯萃取（3×10mL），合并有机层，无水Na2SO4干燥、过滤，减压浓缩，经柱层析纯化得化合物O，称取化合物O（0.38mmol）溶于MeOH/H2O/DMSO（2mL,v/v/v=2:1:1）的混合溶剂中，加入3.8mmolNaOH，室温反应26-30h（TLC监测反应完全），然后加入3mL5%的盐酸溶液，乙酸乙酯萃取，合并有机相，无水Na2SO4干燥、过滤，取滤液减压浓缩得到化合物P。Figure 3 is the technical route for preparing compound P. Specific operation method: weigh compound I (4.03mmol) and dissolve it in 10mL ultra-dry tetrahydrofuran, add triethylamine (4.8mmol, 1.2eq), then add tert-butyl hypochlorite (4.83mmol, 1.2eq), after reacting for 40min, add compound B (8.06mmol, 2.0eq) and boron trifluoride. Ether (16.09mmol, 4.0eq) successively, rise to room temperature and react for 5h, slowly add saturated aqueous sodium bicarbonate solution to adjust pH =7, extracted with ethyl acetate (3×10mL), combined the organic layers, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was separated and purified by column chromatography to obtain compound J; compound J (0.97mmol) Dissolve in methanol/dichloromethane (v/v=1:1) mixed solvent, add 80% hydrazine hydrate (3.88mmol, 4.0eq), react at room temperature overnight, filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, Compound K was obtained by separation and purification by column chromatography; Compound K (0.70 mmol) was dissolved in a mixed solvent of acetonitrile/acetic acid buffer (4 mL, v/v=1:1), and zinc sulfate heptahydrate (0.35 mmol) and Sodium glyoxylate (7.0mmol), react at room temperature for 0.5h, then add 10mL of water to quench the reaction, extract with ethyl acetate (3×10mL), combine organic layers, dry over anhydrous Na2 SO4 , filter, and concentrate under reduced pressure to obtain The crude product is dissolved in 3 mL of 1,4-dioxane, then added with 0.1 mL of trifluoroacetic acid, heated to reflux for 5-8 hours, cooled, concentrated under reduced pressure, separated and purified by column chromatography to obtain Compound L; Dissolve compound L and compound N in 3mL acetone, add potassium carbonate (4.0eq) and tetrabutylammonium iodide (1.2eq), heat to reflux for 5h, evaporate acetone under reduced pressure, add 10mL water, ethyl acetate Ester extraction (3×10 mL), combined organic layers, dried over anhydrous Na2 SO4 , filtered, concentrated under reduced pressure, purified by column chromatography to obtain compound O, weighed compound O (0.38 mmol) and dissolved in MeOH/H2 O /DMSO (2mL, v/v/v=2:1:1) mixed solvent, add 3.8mmol NaOH, react at room temperature for 26-30h (reaction is complete as monitored by TLC), then add 3mL of 5% hydrochloric acid solution, extract with ethyl acetate , the organic phases were combined, dried over anhydrous Na2 SO4 , filtered, and the filtrate was concentrated under reduced pressure to obtain compound P.
实施例5Example 5
C13的制备：Preparation of C13:
制备C13的技术路线为：The technical route for preparing C13 is:
具体操作方法为：将0.20mmol化合物P溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物G，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到黄色固体47.5mg，产率52%。The specific operation method is: dissolve 0.20 mmol of compound P in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound G, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 47.5 mg of a yellow solid with a yield of 52%.
C13的NMR数据见表2。See Table 2 for the NMR data of C13.
实施例6Example 6
C14的制备：Preparation of C14:
制备C14的技术路线为：The technical route for preparing C14 is:
具体操作方法为：将0.20mmol化合物P溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物H，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到黄色固体39.6mg，产率41%。The specific operation method is: dissolve 0.20 mmol of compound P in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound H, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 39.6 mg of a yellow solid with a yield of 41%.
C14的NMR数据见表2。The NMR data of C14 are shown in Table 2.
原料U的制备Preparation of raw material U
图4为制备化合物U的技术路线。具体操作方法：称取化合物A（4.03mmol）溶于10mL超干四氢呋喃中，加入三乙胺（4.8mmol，1.2eq），然后于-78℃下加入次氯酸叔丁基酯（4.83mmol,1.2eq），反应40min后，依次加入化合物Q（8.06mmol,2.0eq）和三氟化硼.乙醚（16.09mmol,4.0eq），升至室温反应5h，缓慢加入饱和的碳酸氢钠水溶液调pH=7，乙酸乙酯萃取（3×10mL），合并有机层，无水硫酸镁干燥，过滤，减压浓缩得粗产品，再经柱层析分离纯化得化合物R；将化合物R（0.97mmol）溶于甲醇/二氯甲烷（v/v=1:1）的混合溶剂中，加入80％的水合肼（3.88mmol,4.0eq），室温反应过夜，过滤，取滤液减压浓缩得粗产品，再经柱层析分离纯化得到化合物S；化合物S（0.70mmol）溶于乙腈/醋酸缓冲液（4mL,v/v=1:1）的混合溶剂中，加入七水合硫酸锌（0.35mmol）和乙醛酸钠（7.0mmol），室温反应0.5h，然后加入10mL水淬灭反应，乙酸乙酯萃取（3×10mL），合并有机层，无水Na2SO4干燥、过滤、减压浓缩得粗产物，将此粗产物溶于3mL的1,4-二氧六环中，然后加入0.1mL三氟乙酸，加热回流反应5-8h，冷却后，减压浓缩，经柱层析分离纯化得到目化合物T；称取化合物T（0.38mmol）溶于MeOH/H2O/DMSO（2mL,v/v/v=2:1:1）的混合溶剂中，加入3.8mmolNaOH，室温反应26-30h（TLC监测反应完全），然后加入3mL5%的盐酸溶液，乙酸乙酯萃取，合并有机相，无水Na2SO4干燥、过滤，取滤液减压浓缩得到化合物U。Figure 4 is the technical route for preparing compound U. Specific operation method: weigh compound A (4.03mmol) and dissolve it in 10mL ultra-dry tetrahydrofuran, add triethylamine (4.8mmol, 1.2eq), then add tert-butyl hypochlorite (4.83mmol, 1.2eq), after reacting for 40min, add compound Q (8.06mmol, 2.0eq) and boron trifluoride. Ether (16.09mmol, 4.0eq) successively, rise to room temperature and react for 5h, slowly add saturated aqueous sodium bicarbonate solution to adjust pH =7, extracted with ethyl acetate (3×10mL), combined the organic layers, dried over anhydrous magnesium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product, which was separated and purified by column chromatography to obtain compound R; compound R (0.97mmol) Dissolve in methanol/dichloromethane (v/v=1:1) mixed solvent, add 80% hydrazine hydrate (3.88mmol, 4.0eq), react at room temperature overnight, filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, Compound S was obtained by separation and purification by column chromatography; Compound S (0.70 mmol) was dissolved in a mixed solvent of acetonitrile/acetic acid buffer (4 mL, v/v=1:1), and zinc sulfate heptahydrate (0.35 mmol) and Sodium glyoxylate (7.0mmol), react at room temperature for 0.5h, then add 10mL of water to quench the reaction, extract with ethyl acetate (3×10mL), combine organic layers, dry over anhydrous Na2 SO4 , filter, and concentrate under reduced pressure to obtain The crude product is dissolved in 3 mL of 1,4-dioxane, then added with 0.1 mL of trifluoroacetic acid, heated to reflux for 5-8 hours, cooled, concentrated under reduced pressure, separated and purified by column chromatography to obtain Target compound T; weigh compound T (0.38mmol) and dissolve it in a mixed solvent of MeOH/H2 O/DMSO (2mL, v/v/v=2:1:1), add 3.8mmolNaOH, and react at room temperature for 26-30h (TLC monitors that the reaction is complete), then add 3 mL of 5% hydrochloric acid solution, extract with ethyl acetate, combine the organic phases, dry over anhydrous Na2 SO4 , filter, and concentrate the filtrate under reduced pressure to obtain compound U.
实施例7Example 7
C15的制备：Preparation of C15:
制备C15的技术路线为：The technical route for preparing C15 is:
具体操作方法为：将0.2mmol化合物U溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物G，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到白色固体64.9mg，产率为81%;The specific operation method is: dissolve 0.2 mmol of compound U in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound G, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 64.9 mg of white solid with a yield of 81%;
C15的NMR数据见表2。See Table 2 for the NMR data of C15.
实施例8Example 8
C16的制备：Preparation of C16:
制备C16的技术路线为：The technical route for preparing C16 is:
具体操作方法为：将0.2mmol化合物U溶于干燥的DMF中，加入0.25mmolHOBt·H2O和0.38mmolEDCI，室温搅拌20min后再依次加入0.5mmolEt3N和0.25mmol化合物H，室温搅拌过夜。向反应体系中加入20mL饱和Na2CO3，乙酸乙酯萃取两次（2×10mL），合并有机相，水洗，无水Na2SO4干燥、过滤，取滤液减压浓缩得粗产品，再经柱层析纯化得到黄色固体72.5mg，产率为85%;The specific operation method is: dissolve 0.2 mmol of compound U in dry DMF, add 0.25 mmol of HOBt·H2 O and 0.38 mmol of EDCI, stir at room temperature for 20 min, then add 0.5 mmol of Et3 N and 0.25 mmol of compound H, and stir overnight at room temperature. Add 20 mL of saturated Na2 CO3 to the reaction system, extract twice with ethyl acetate (2×10 mL), combine the organic phases, wash with water, dry over anhydrous Na2 SO4 , filter, take the filtrate and concentrate under reduced pressure to obtain the crude product, and then Purified by column chromatography to obtain 72.5 mg of a yellow solid with a yield of 85%;
C16的NMR数据见表2.The NMR data of C16 are shown in Table 2.
实施例9Example 9
C27的制备：Preparation of C27:
具体操作方法为：称取50mgC13（0.11mmol）溶于2mLCH2Cl2，加入0.5mL二氧六环的盐酸溶液，室温搅拌30min，旋干溶剂便得到黄色固体54mg，产率100%。The specific operation method is: weigh 50mg of C13 (0.11mmol) and dissolve it in 2mL of CH2 Cl2 , add 0.5mL of dioxane hydrochloric acid solution, stir at room temperature for 30min, and spin the solvent to obtain 54mg of a yellow solid with a yield of 100%.
实施例10Example 10
C28的制备：Preparation of C28:
具体操作方法为：称取50mgC14（0.10mmol）溶于2mLCH2Cl2，加入0.5mL二氧六环的盐酸溶液，室温搅拌30min，旋干溶剂便得到黄色固体57.4mg，产率100%。The specific operation method is: weigh 50mg of C14 (0.10mmol) and dissolve it in 2mL of CH2 Cl2 , add 0.5mL of dioxane hydrochloric acid solution, stir at room temperature for 30min, and spin the solvent to obtain 57.4mg of a yellow solid with a yield of 100%.
以下通过药效学试验和对比试验来进一步阐述本发明所述药物的有益效果。The beneficial effect of the medicine of the present invention will be further elaborated below through pharmacodynamic tests and comparative tests.
本发明实验使用的肿瘤细胞株人红白血病细胞HEL，骨髓性白血病细胞K562-1，人黑素瘤细胞WM9-1，乳腺癌细胞MDA231，B淋巴母细胞TK-6，FV-P诱导红白血病(小鼠细胞)DP17-17，前列腺癌细胞PC-3为上海细胞库提供。The tumor cell lines used in the experiment of the present invention are human erythroleukemia cells HEL, myeloid leukemia cells K562-1, human melanoma cells WM9-1, breast cancer cells MDA231, B lymphoblastoid cells TK-6, and FV-P to induce erythroleukemia (Mouse cells) DP17-17 and prostate cancer cell PC-3 were provided by Shanghai Cell Bank.
实施例11Example 11
采用MTT法测定优选的C1、C3、C11、C12、C13、C14、C15、C16、C27、C28这十种化合物的抗肿瘤活性。The antitumor activity of the preferred ten compounds C1, C3, C11, C12, C13, C14, C15, C16, C27, and C28 was determined by MTT method.
具体方法：取对数生长期的人人红白血病细胞HEL，骨髓性白血病细胞K562-1，人黑素瘤细胞WM9-1，乳腺癌细胞MDA231，B淋巴母细胞TK-6，FV-P诱导红白血病(小鼠细胞)DP17-17，前列腺癌细胞PC-3，分别以2×104个/mL细胞密度接种于96孔培养板中，100μL/孔，每种细胞各种4块板。置37℃，5％CO2孵箱内培养12h。吸弃上清，然后分别加入200μL不同浓度的待测化合物，同时设阳性对照药组和空白细胞对照组，每组设4个复孔。培养72h后，再加入5mg/mL的MTT20μL/孔，继续培养4h后弃上清，加入DMSO150μL/孔，微振荡器上振荡10min，将试剂对照调零，用自动酶标仪在550nm波长处测出细胞对照组和各药物组的OD值，取各组均值，重复实验3次。以下面的公式计算各组药物对细胞的抑制率IR＝(1-药物组OD值/细胞对照组OD值)×100％，同时计算IC50值（实验结果见表3）。从表3可以看出优选出的化合物对HEL（人红白血病细胞），K562-1（骨髓性白血病细胞），WM9-1（人黑素瘤细胞），MDA231（乳腺癌细胞），TK-6（B淋巴母细胞），DP17-17（FV-P诱导红白血病(小鼠细胞)），PC-3（前列腺癌细胞）都有比较好的活性。Specific method: human erythroleukemia cells HEL in logarithmic growth phase, myeloid leukemia cells K562-1, human melanoma cells WM9-1, breast cancer cells MDA231, B lymphoblastoid cells TK-6, FV-P induction Erythroleukemia (mouse cells) DP17-17 and prostate cancer cells PC-3 were inoculated in 96-well culture plates at a cell density of 2×104/mL, 100 μL/well, 4 plates for each type of cell. Place them in a 37°C, 5% CO2 incubator for 12 hours. The supernatant was discarded, and then 200 μL of different concentrations of the compound to be tested were added respectively, and a positive control drug group and a blank cell control group were set at the same time, and 4 replicate wells were set for each group. After culturing for 72 hours, add 5 mg/mL MTT 20 μL/well, continue to cultivate for 4 hours, discard the supernatant, add DMSO 150 μL/well, vibrate on a micro-oscillator for 10 minutes, set the reagent control to zero, and use an automatic microplate reader to measure at a wavelength of 550 nm. The OD values of the cell control group and each drug group were obtained, and the mean value of each group was taken, and the experiment was repeated 3 times. Use the following formula to calculate the inhibitory rate IR of each group of drugs on the cells = (1-OD value of the drug group/OD value of the cell control group) × 100%, and calculate the IC50 value at the same time (see Table 3 for the experimental results). It can be seen from Table 3 that the preferred compounds are effective against HEL (human erythroleukemia cells), K562-1 (myeloid leukemia cells), WM9-1 (human melanoma cells), MDA231 (breast cancer cells), TK-6 (B lymphoblastoid cells), DP17-17 (FV-P induced erythroleukemia (mouse cells)), PC-3 (prostate cancer cells) have relatively good activity.
对C1进行了体外体内抗肿瘤的实验：In vitro and in vivo anti-tumor experiments were carried out on C1:
实施例12Example 12
用台盼蓝细胞计数法检测C1作用3days对K562细胞生长的影响，具体方法用酒精清洁计数板和盖玻片，然后用吸水纸轻轻擦干，用胰蛋白酶消化单层培养细胞或收集悬浮培养细胞，制成单个细胞悬液，将盖玻片盖在计数板两槽中间，用吸管轻轻吹打细胞悬液，吸取少量细胞悬液与等比例台盼蓝染色液混合，在计数板上盖玻片一侧加细胞悬液，在显微镜下，用10倍物镜观察计数板四角大方格中的细胞数，计数未被染色的细胞，细胞压中线时，只计左侧和上方者，不计右侧和下方者。Use the trypan blue cell counting method to detect the effect of C1 on the growth of K562 cells for 3 days. The specific method is to clean the counting plate and cover slip with alcohol, then dry it gently with absorbent paper, and digest the monolayer culture cells with trypsin or collect the suspension Cultivate the cells to make a single cell suspension, cover the cover glass between the two grooves of the counting plate, gently blow the cell suspension with a pipette, draw a small amount of the cell suspension and mix it with an equal proportion of trypan blue staining solution, and put it on the counting plate Add cell suspension to one side of the cover glass, observe the number of cells in the large squares at the four corners of the counting plate under a microscope with a 10X objective lens, and count the unstained cells. When the cells press the midline, only the left and upper ones are counted. Those on the right and below are not counted.
图5为C1对K562细胞生长的影响曲线图；从中可以看出化合物C1的浓度与对细胞的抑制率呈正相关，表现出对K562细胞有很好的抑制作用。Figure 5 is a graph showing the effect of C1 on the growth of K562 cells; it can be seen that the concentration of compound C1 is positively correlated with the inhibitory rate on cells, showing a good inhibitory effect on K562 cells.
实施例13Example 13
用流式细胞术检测C1作用72h对K562细胞周期的影响。The effect of C1 on the K562 cell cycle was detected by flow cytometry for 72 hours.
图6为C1作用72h时对K562细胞细胞周期的影响，从中可以看出C1对K562细胞细胞周期有明显的影响，阻断在G2期。Figure 6 shows the effect of C1 on the cell cycle of K562 cells when C1 acts for 72 hours, from which it can be seen that C1 has a significant effect on the cell cycle of K562 cells, blocking in the G2 phase.
实施例14Example 14
给刚出生一天的小鼠注射Friendvirus病毒，35天后连续用C1药（1mg/kg)两周，然后解剖，测其脾脏重量。用微量毛细管比容法测红细胞比容，具体方法是以抗凝剂湿润毛细管内壁后吹出，让壁内自然风干或于60℃～80℃干燥箱内干燥后待用；取血：常规消毒，吸取心脏血，使血液进入毛细管的2/3（约50mm）处；离心：用酒精灯溶封或橡皮泥、石蜡封堵其未吸血端，然后封端向外放入专用的水平式毛细管离心机，以12000r/min的速度离心5min，用刻度尺分别量出红细胞柱和全血柱高度（单位mm）。计算其比值，即得出红细胞比容。Inject Friendvirus virus to the mouse of just one day after birth, and use C1 medicine (1mg/kg) continuously for two weeks after 35 days, then dissect, measure its spleen weight. Measure the hematocrit by the microcapillary specific volume method. The specific method is to wet the inner wall of the capillary with anticoagulant and blow it out, let the wall dry naturally or dry it in a drying oven at 60°C to 80°C before use; blood collection: routine disinfection, Draw blood from the heart to make the blood enter the 2/3 (about 50mm) of the capillary; centrifuge: use an alcohol lamp to melt seal or plasticine, paraffin to seal the non-absorbing end, and then seal the end outward and put it into a special horizontal capillary centrifuge Centrifuge at a speed of 12000r/min for 5min, and use a scale to measure the height of the red blood cell column and the whole blood column (in mm). Calculate its ratio, that is, the hematocrit.
图7为C1（1mg/kg)给药两周对感染F-MuLV小鼠脾脏重量的影响，一般的正常脾脏重量为0.1左右，而患病的则为0.4左右，从图5可以看出C1对患病的小鼠具有很明显的治疗作用；Fig. 7 is the impact of C1 (1mg/kg) administration on the spleen weight of infected F-MuLV mice for two weeks, the general normal spleen weight is about 0.1, and the diseased one is about 0.4, as can be seen from Fig. 5 It has obvious therapeutic effect on sick mice;
图8为C1（1mg/kg)给药两周对感染F-MuLV小鼠红细胞比容的影响，一般正常的红细胞比容为50%，患病的红细胞比容为25%左右，从图中均可以看出C1对Friendvirus诱发的白血病有治疗作用。Fig. 8 is the influence of C1 (1mg/kg) administration two weeks on the hematocrit of infected F-MuLV mice, the general normal hematocrit is 50%, and the sick hematocrit is about 25%, from the figure It can be seen that C1 has a therapeutic effect on leukemia induced by Friendvirus.
上述实验结果表明：吲哚并[3,2-a]咔唑类化合物或其药学上可接受的盐对不同的肿瘤细胞株均显示显著的抑制作用，因此可以用于制备抗肿瘤药物。本发明为研制抗肿瘤药物提供了新的化学实体或先导化合物，为开发利用中国的传统药物具有重要意义。The above experimental results show that the indolo[3,2-a]carbazole compounds or their pharmaceutically acceptable salts all exhibit significant inhibitory effects on different tumor cell lines, and therefore can be used to prepare antitumor drugs. The invention provides a new chemical entity or lead compound for the development of antitumor drugs, and is of great significance for the development and utilization of traditional Chinese medicines.
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| CN201510893281.2ACN105693729B (en) | 2015-12-08 | 2015-12-08 | Indoles simultaneously [3,2 a] carbazole derivates and its application |
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| CN110041338A (en)* | 2018-10-30 | 2019-07-23 | 贵州省中国科学院天然产物化学重点实验室 | One kind double indoles maleic amide class compounds with anti-tumor activity and its application |
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| CN113004272B (en)* | 2021-03-01 | 2023-06-16 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Quinoline [4,3-b ] carbazole derivative and application thereof in preparation of antitumor drugs |
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