CN116655510A - 一种四氢吡咯烷类化合物及其制备方法和应用 - Google Patents
一种四氢吡咯烷类化合物及其制备方法和应用 Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D207/00—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom
- C07D207/02—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom
- C07D207/04—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members
- C07D207/10—Heterocyclic compounds containing five-membered rings not condensed with other rings, with one nitrogen atom as the only ring hetero atom with only hydrogen or carbon atoms directly attached to the ring nitrogen atom having no double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
- C07D207/16—Carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
- C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
- C07D405/04—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
- C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
- C07D409/04—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings directly linked by a ring-member-to-ring-member bond
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Abstract
发明属于肿瘤药物技术领域,具体涉及一种四氢吡咯烷类化合物及其制备方法和应用。该四氢吡咯烷类化合物如式(I)所示,其具有良好的抗肿瘤作用,对正常细胞的毒性低,是安全、有效和低毒的候选抗肿瘤药物。
Description
技术领域
本发明属于肿瘤药物技术领域,具体涉及一种四氢吡咯烷类化合物及其制备方法和应用。
背景技术
恶性肿瘤是目前全世界人类的主要死亡原因之一,已经成为严重危害人类生命健康、制约社会经济发展的一大类疾病。目前的治疗方法有化疗、放疗、手术等,但仍主要以化疗为主,治疗恶性肿瘤的化学药物有多类化合物,如氮芥、嘧啶类、铂类、卟啉类等,但大多数药物由于毒性大、生物利用度低而使其应用受到了限制。因此,寻找高效、低毒的抗肿瘤药物已成为当前药物化学领域研究热点之一。
含有手性季碳中心的四氢吡咯环是众多具有重要生物活性的药物分子和天然产物的重要结构单元,在医药领域有着广泛的应用,主要表现在抗菌、抗肿瘤、神经疾病和心血管等方面,因其具有的低毒和高效的特性而倍受重视。由于该类四氢吡咯环含有手性季碳中心,环上引入不同的取代基会使分子的构象具有很大的多变性,可能会改善原有的理化性质和药理活性,在临床应用上有极其重要的作用。
因此,挖掘一种新的含手性季碳中心的四氢吡咯烷类化合物是有必要的。
发明内容
针对以上问题,本发明目的之一在于提供一种含手性季碳中心的四氢吡咯烷类化合物,该化合物是在双功能化配体在金属的搭配下,金属催化和有机催化协同促进的α取代丙烯腈和多取代硝基乙烯作为亲偶极体与α取代或无取代亚胺酯的1,3-偶极环加成反应制备而得。该化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物具有良好的抗肿瘤作用,对正常细胞的毒性低,是安全、有效和低毒的候选抗肿瘤药物。
为了达到上述目的,本发明可以采用以下技术方案:
本发明一方面提供一种式(I)的四氢吡咯烷类化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物:
其中,R1任意选自:
R2和R3独立任意选自:R2和R3相同或不同;R4任意选自:/>
R1和R4相同或不同;
R5任意选自:
R2、R3和R5相同或不同。
本发明另一方面提供一种上述式(I)所示的四氢吡咯烷类化合物的制备方法,其包括:将配体L1、分子筛、Cu(CH3CN)4BF4和二氯甲烷混合反应后降温至0℃±5℃得反应液;式(II)所示化合物、式(III)所示化合物、三乙胺和二氯甲烷混合,然后与反应液混合进行反应;反应结束后得式(I)所示化合物;
本发明再一方面提供一种药物组合物,其包括上述的化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
本发明再一方面提供一种制剂,其包括上述化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物;或上述的药物组合物;和可药用载体。
本发明再一方面提供一种上述的化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物;或上述的药物组合物在作为或制备抗癌细胞活性的制剂中的应用。
本发明有益效果包括:本发明提供的四氢吡咯烷类化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物具有良好的抗肿瘤作用,对正常细胞的毒性低,是安全、有效和低毒的候选抗肿瘤药物。
具体实施方式
所举实施例是为了更好地对本发明进行说明,但并不是本发明的内容仅局限于所举实施例。所以熟悉本领域的技术人员根据上述发明内容对实施方案进行非本质的改进和调整,仍属于本发明的保护范围。
本文中使用的术语仅用于描述特定实施例,并且无意于限制本公开。除非在上下文中具有明显不同的含义,否则单数形式的表达包括复数形式的表达。如本文所使用的,应当理解,诸如“包括”、“具有”、“包含”之类的术语旨在指示特征、数字、操作、组件、零件、元件、材料或组合的存在。在说明书中公开了本发明的术语,并且不旨在排除可能存在或可以添加一个或多个其他特征、数字、操作、组件、部件、元件、材料或其组合的可能性。如在此使用的,根据情况,“/”可以被解释为“和”或“或”。
本发明实施例提供一种式(I)的四氢吡咯烷类化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物:
其中,R1任意选自:
R2和R3独立任意选自:R2和R3相同或不同;
R4任意选自:
R1和R4相同或不同;
R5任意选自:
R2、R3和R5相同或不同。
在一些具体实施例中,
R1任意优选自:和/或
R2任意优选自:和/或
R3任意优选自:和/或
R4任意优选自:和/或
R5任意选自:
需要说明的是,上述式(I)的四氢吡咯烷类化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物具有杀伤癌症细胞的能力,而且细胞毒性低。另外,其中的立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物为本领域所常规的技术术语,没有其他特指含义。
在一些具体实施例中,上述式(I)的四氢吡咯烷类化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物可以任意优选自:
(2S,3R,4S,5S)-2,4-二甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氟苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氯苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-溴苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-甲氧基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(苯并[d][1,3]二恶英-5-基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-甲基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-三氟甲基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-硝基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氰基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(2-氟苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(3-氟苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(3-溴苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-2,4-二甲基-5-(萘-1-基)-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-2,4-二甲基-5-(萘-2-基)-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-5-(噻吩-2-基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-5-(呋喃-2-基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-溴苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-氯苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-氟苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-硝基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-甲氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(呋喃-2-基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-4-乙基-2-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-2,4-二甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸乙酯;
(2S,3R,4S,5S)-2,4-二甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸叔丁酯;
(2S,3R,4S,5S)-2-乙基-4-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-4-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3S,4S,5S)-2-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氯苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-甲氧基苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-三氟甲基苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-(噻吩-2-基)吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-乙氧基苯基)-4-甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3S,4S,5S)-3-(4-乙氧基苯基)-2-甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
在一些具体实施例中,上述式(I)的四氢吡咯烷类化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物可以任意优选自:
(2S,3R,4S,5S)-3-(4-甲氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氯苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-(噻吩-2-基)吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-乙氧基苯基)-4-甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
需要说明的是,上述式(I)化合物药学上可接受的盐包括但不限于各种无机或有机酸盐如盐酸盐、氢溴酸盐、磷酸盐、硫酸盐、柠檬酸盐、乳酸盐、酒石酸盐、马来酸盐、延胡索酸盐、扁桃酸盐和草酸盐;各种无机或有机碱盐如氢氧化钠、三羟甲基氨基甲烷和N-甲基-葡萄糖胺。
还需要说明的是,上述优选的式(I)化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物较其他的化合物对癌症细胞有较强的杀伤能力;而且细胞毒性低。
本发明另一实施例提供一种上述式(I)所示的四氢吡咯烷类化合物的制备方法,其包括:将配体L1、分子筛、Cu(CH3CN)4BF4和二氯甲烷混合反应后降温至0℃±5℃得反应液;式(II)所示化合物、式(III)所示化合物、三乙胺和二氯甲烷混合,然后与反应液混合进行反应;反应结束后得式(I)所示化合物;
需要说明的是,上述制备方法中,式(II)所示化合物和式(III)所示化合物的R1、R2、R3、R4和R5与上述式(I)所示的四氢吡咯烷类化合物中的R1、R2、R3、R4和R5一致。
本发明再一实施例提供一种药物组合物,其包括上述的式(I)化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
需要说明的是,可以将上述的式(I)化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物与其他活性药物形成复方联合使用,增强对癌症细胞的杀伤能力或者补充其他疗效达成协同作用。
还需要说明说明的是,本发明的药物组合物有0.1%-99.9%重量百分比的作为活性成分的上述的式(I)化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
本发明再一方面提供一种制剂,其包括上述式(I)化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物;或上述的药物组合物;和可药用载体。
需要说明的是,上述制剂中,可药用载体包括但不限于:离子交换材料、氧化铝、硬脂酸铝、卵磷脂、自乳化药物传递系统(SEDDS)如d-维生素E聚乙二醇1000琥珀酸酯、吐温或其他类似聚合介质等药物制剂用的表面活性剂、血清蛋白如人血清白蛋白、缓冲物质如磷酸盐、氨基乙酸、山梨酸、山梨酸钾、饱和植物脂肪酸部分甘油酯混合、水、盐、电解质如硫酸盐精蛋白、磷酸氢二钠、磷酸氢钾、氯化钠、锌盐、硅胶、硅酸镁等。聚乙烯吡咯酮、纤维素物质、聚乙烯醇、羧甲基纤维素钠、聚丙烯酸酯、乙烯-聚氧乙烯-嵌段聚合物和羊毛脂、环糊精如α-、β-、γ-环糊精或其经化学修饰的衍生物如2-和3-羟丙基-β-环糊精等羟烷基环糊精或其他可溶性衍生物等均可用于促进上述式(I)所示化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物的药物传递。
还需说明的是,上述制剂中,可药用载体还包括可药用辅料,如填充剂(如无水乳糖、淀粉、乳糖珠粒和葡萄糖)、粘合剂(如微晶纤维素)、崩解剂(如交联羧甲基淀粉钠、交联羧甲基纤维素钠、低取代羟丙基纤维素和交联PVP)、润滑剂(如硬脂酸镁)、吸收促进剂、香味剂、甜味剂、稀释剂、赋形剂、润湿剂、溶剂、增溶剂和着色剂等也可以作为可药用载体用于将上述化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物制备成制剂。
在一些具体实施例中,上述制剂中,可药用载体适用于液体剂型、固体剂型或膏体剂型。
需要说明的是,可以根据上述式(I)所示化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物的给药方式,选择适用于不同剂型的可药用载体。还需要说明的是,液体剂型、固体剂型、膏体剂型或乳状剂型为本领域所已知,比如液体剂型包括但不限于:注射剂、喷雾剂或口服液等剂型;固体剂型包括但不限于:片剂、粉剂、颗粒剂或胶囊剂等;膏体剂型包括但不限于:霜剂或软膏剂等。在一些具体实施中,上述式(I)所示化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物可通过肠道或者非肠道途径给药;非肠道给药制剂包括注射剂、霜剂、软膏剂、贴剂、喷雾剂等;给药途径包括皮下、皮内、动脉内、静脉内、肌内、关节内、滑液内、胸骨内、鞘内、病灶内、颅内注射或输注,或者,口服、局部、直肠、经鼻、经颊、阴道、舌下、皮内、粘膜、气管、尿道给药,或者通过吸入气雾或植入蓄积或者针刺方式给药。
本发明再一方面提供一种上述的式(I)化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物;或上述的药物组合物在作为或制备抗癌细胞活性的制剂中的应用。
在一些具体实施例中,上述应用中,癌细胞为本领域所已知的癌细胞,优选黑色素瘤细胞、肺腺癌细胞、结肠癌细胞、乳腺癌细胞、肝癌细胞或宫颈癌细胞等。
需要说明的是,上述式(I)所示化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物的治疗有效量为0.001mg/kg/d-100mg/kg/d之间,可用于相关疾病的单一用药或联合用药治疗,为本领域技术人员能够理解的范围。
为了更好地理解本发明,下面结合具体示例进一步阐明本发明的内容,但本发明的内容不仅仅局限于下面的示例。
以下实施例中,含手性季碳中心的四氢吡咯烷类化合物按照以下合成路线进行合成:
以下实施例中,按照上述合成路线制备含手性季碳中心的四氢吡咯烷类化合物的方法包括:
(1)化合物3制备
称取化合物2(1.5eq)和无水硫酸镁(1.5eq)于干燥的圆底烧瓶中,加入搅拌子,氮气保护,加入干燥的CH2Cl2作溶剂,加入Et3N(1.5eq),室温搅拌1h,然后加入化合物1(1eq),室温搅拌24h,待反应完全后将反应液过滤,滤液用去离子水配制的饱和氯化钠溶液,饱和碳酸氢钠溶液依次洗2次,最后用纯水洗涤2次,合并有机相,用无水硫酸钠干燥,真空减压浓缩,得到化合物3粗产物,粗产物无需纯化,直接用于下一步反应。
(2)化合物6制备
称取化合物4(1eq),化合物5(用作溶剂),醋酸铵(1.3eq)于圆底烧瓶内,120℃回流反应2小时。待反应完全后加水淬灭反应,乙酸乙酯萃取三次,合并有机相,用食盐水洗一次,无水硫酸钠干燥,真空浓缩,硅胶柱色谱纯化得到产物化合物6。
(3)化合物I制备
称取配体L1(0.011mmol,0.055eq)与Cu(CH3CN)4BF4(0.01mmol,0.05eq)于干燥反应试管中,加入分子筛(200mg),加入搅拌子,氮气保护,加入2.5mL干燥的二氯甲烷,置于室温下搅拌30min,移至0℃,将(2)中制备的化合物3(0.22mmol,1.1eq)、(2)中制备的化合物6(0.2mmol,1eq)和三乙胺(0.1mmol,0.5eq)分别溶于0.5mL干燥的二氯甲烷加入反应液中,0℃进行反应,TLC监测反应进程;待反应完全后,过滤掉分子筛,滤液依次用饱和碳酸氢钠溶液,饱和氯化钠溶液,用无水硫酸钠干燥有机相,真空减压浓缩,粗产品经硅胶柱层析纯化得到目标产物化合物I。
一、化合物I的制备及表征
(1)化合物I的制备
实施例1至实施例36分别利用下表1所示的不同的原料按照上述制备方法制备化合物I。
表1实施例1至实施例36使用的原料
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(2)化合物I的表征
实施例1至实施例36制备的化合物I的名称、结构式、产率以及氢核磁共振数据如下表2所示。
表2实施例1至实施例36制备的化合物I表征数据
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二、化合物I体外抗肿瘤活性以及细胞毒性测定
使用MTT[3-(4,5)-双甲基-2-噻唑-(2,5)-苯基溴化四氮唑蓝]法来分别测定实施例1至实施例36制备得到的化合物I分别对人黑色素瘤细胞株(A375)、人肺腺癌细胞(A549)和人结肠癌细胞株(HCT116)的抑制率达到50%时的药物浓度(half maximal inhibitoryconcentration,IC50);以及对人肝细胞(L02)的细胞毒性,每个化合物的毒性用抑制L02细胞存活率到50%时的浓度CC50来表示。
具体步骤如下:
(1)培养液的配制:DMEM(基础培养基)89%,胎牛血清10%,青霉素链霉素溶液(10000IU/mL,10000μg/mL)1%;
(2)四种细胞的培养:利用(a)中配制的培养液(培养液体积约为培养瓶容量的1/10),在37℃,5% CO2培养箱中分别培养人黑色素瘤细胞株(A375)、人肺腺癌细胞(A549)、人结肠癌细胞株(HCT116)和人肝细胞(L02),根据细胞的生长状态判断传代时间;
(3)不同浓度药物的配制:利用DMEM(少量DMSO助溶)配制储备液,加药后的每孔细胞悬液中DMSO的终浓度一般不超过0.05%-0.1%;用DMEM把储备液稀释至六个浓度梯度(100μM、50μM、10μM、1μM、0.1μM和0.01μM),保存于-20℃冰箱中备用;
(4)细胞孵育:取对数生长期肿瘤细胞,调细胞悬液浓度为5×104/mL,混匀后加入96孔培养板中(100μL/孔),在37℃,5% CO2培养箱中培养24h;
(5)加药:将稀释好的不同浓度梯度的药物分别加入到96孔培养板中,每个浓度梯度设3个复孔,继续培养72h;实验分为实验组(培养液、细胞和化合物I)、对照组(培养液和细胞)和空白组(只有培养液);
(6)存活细胞检测:在培养了72h后的96孔板中,加MTT(5mg/mL)20μL/孔;在37℃放置4h后,移除上清液,加DMSO 200μL/孔,振荡至甲瓒结晶全部溶解;利用自动酶标仪在570nm波长处检测各孔的光密度(OD值)。
(7)抑制率的计算:利用以下公式分别计算实施例1至实施例36制备的化合物I对癌细胞(A375、A549和HCT116)杀伤能力和对正常细胞(L02)的毒性。
生长抑制率=(1-存活率)×100%=[1-(OD实验-OD空白)/(OD对照-OD空白)]×100%,其中,OD实验为实验组的平均光密度,OD对照为对照组的平均光密度,OD空白为空白组的平均光密度。
(8)根据化合物I浓度-不同癌细胞(A375、A549和HCT116)生长抑制率,求其IC50,单位μM;根据化合物I浓度-正常细胞(L02)生长抑制率,求其CC50,单位μM;结果如下表3所示。
表3实施例1至实施例36制备的化合物I的癌细胞杀伤能力和正常细胞毒性数据
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注:表3中,IC50:半数抑制浓度,化合物杀伤癌细胞能力;CC50:半数毒性浓度,化合物对正常细胞的毒性。
以上结果表明,实施例1至实施例36制备的含手性季碳中心的四氢吡咯烷类化合物(化合物I)均具有抗肿瘤作用,对正常细胞的毒性低。
最后说明的是,以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的宗旨和范围,其均应涵盖在本发明的权利要求范围。
Claims (10)
1.式(I)所示的四氢吡咯烷类化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物:
其中,R1任意选自:
R2和R3独立任意选自:R2和R3相同或不同;
R4任意选自:
R1和R4相同或不同;
R5任意选自:
R2、R3和R5相同或不同。
2.根据权利要求1所述的化合物,其特征在于,
R1任意优选自:和/或
R2任意优选自:和/或
R3任意优选自:和/或
R4任意优选自:和/或
R5任意选自:
3.根据权利要求1所述的化合物,其特征在于,任意优选自:
(2S,3R,4S,5S)-2,4-二甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氟苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氯苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-溴苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-甲氧基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;(2S,3R,4S,5S)-5-(苯并[d][1,3]二恶英-5-基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-甲基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-三氟甲基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;(2S,3R,4S,5S)-5-(4-硝基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-氰基苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(2-氟苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(3-氟苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(3-溴苯基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-2,4-二甲基-5-(萘-1-基)-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-2,4-二甲基-5-(萘-2-基)-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-5-(噻吩-2-基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-5-(呋喃-2-基)-2,4-二甲基-4-硝基-3-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-溴苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-氯苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-氟苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-硝基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-甲氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;(2S,3R,4S,5S)-3-(呋喃-2-基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-4-乙基-2-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-2,4-二甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸乙酯;
(2S,3R,4S,5S)-2,4-二甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸叔丁酯;
(2S,3R,4S,5S)-2-乙基-4-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-4-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3S,4S,5S)-2-甲基-4-硝基-3,5-二苯基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;(2S,3R,4S,5S)-5-(4-氯苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-甲氧基苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-5-(4-三氟甲基苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-(噻吩-2-基)吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-乙氧基苯基)-4-甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
(2S,3S,4S,5S)-3-(4-乙氧基苯基)-2-甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
4.根据权利要求1所述的化合物,其特征在于,任意优选自:
(2S,3R,4S,5S)-3-(4-甲氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;(2S,3R,4S,5S)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;(2S,3R,4S,5S)-5-(4-氯苯基)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5R)-3-(4-乙氧基苯基)-2,4-二甲基-4-硝基-5-(噻吩-2-基)吡咯烷-2-羧酸甲酯;
(2S,3R,4S,5S)-3-(4-乙氧基苯基)-4-甲基-4-硝基-5-苯基吡咯烷-2-羧酸甲酯;
及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
5.权利要求1至4中任一权利要求所述的式(I)所示化合物的制备方法,其特征在于,包括:将配体L1、分子筛、Cu(CH3CN)4BF4和二氯甲烷混合反应后降温至0℃±5℃得反应液;式(II)所示化合物、式(III)所示化合物、三乙胺和二氯甲烷混合,然后与反应液混合进行反应;反应结束后得式(I)所示化合物;
6.药物组合物,其特征在于,包括权利要求1至4中任一权利要求所述的化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物。
7.制剂,其特征在于,包括要求1至4中任一权利要求所述的化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物;或权利要求6所述的药物组合物;和可药用载体。
8.根据权利要求7所述的制剂,其特征在于,可药用载体适用于液体剂型、固体剂型或膏体剂型。
9.权利要求1至4中任一权利要求所述的化合物及其药学上可接受的盐、立体异构体、互变异构体、同系物、溶剂化物、前药或多晶型物;或权利要求6所述的药物组合物在作为或制备抗癌细胞活性的制剂中的应用。
10.根据权利要求9所述的应用,其特征在于,癌细胞包括黑色素瘤细胞、人肺腺癌细胞或人结肠癌细胞。
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