CN101495485B - 具有抗肿瘤活性的喜树碱衍生物 - Google Patents
具有抗肿瘤活性的喜树碱衍生物 Download PDFInfo
- Publication number
- CN101495485B CN101495485B CN2007800281978A CN200780028197A CN101495485B CN 101495485 B CN101495485 B CN 101495485B CN 2007800281978 A CN2007800281978 A CN 2007800281978A CN 200780028197 A CN200780028197 A CN 200780028197A CN 101495485 B CN101495485 B CN 101495485B
- Authority
- CN
- China
- Prior art keywords
- compound
- group
- nmr
- cdcl
- nsc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
- C07D491/22—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains four or more hetero rings
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
- A61K31/47—Quinolines; Isoquinolines
- A61K31/4738—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems
- A61K31/4745—Quinolines; Isoquinolines ortho- or peri-condensed with heterocyclic ring systems condensed with ring systems having nitrogen as a ring hetero atom, e.g. phenantrolines
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Oncology (AREA)
- Epidemiology (AREA)
- Hematology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Nitrogen Condensed Heterocyclic Rings (AREA)
- Saccharide Compounds (AREA)
Abstract
具有抗肿瘤活性的5-取代的喜树碱衍生物,其制备方法,其作为抗肿瘤药物的用途和含有它们的药物组合物。
Description
本发明涉及具有抗肿瘤活性的新的喜树碱衍生物、它们的制备方法、其作为抗肿瘤药物的用途以及包含它们的药物组合物。
发明背景
喜树碱是从喜树(Camptotheca acuminata)(Nyssaceae)提取的生物碱,首先由Wall和Wani在1966年描述(J.Am.Chem.Soc.1966,88,3888-3890)。喜树碱尽管具有广谱抗肿瘤活性,特别是抗结肠肿瘤及其他实体肿瘤和白血病的活性,但由于其特别地表现为出血性膀胱炎、胃肠道毒性和骨髓抑制的高毒性而未用于治疗中。
已合成了许多喜树碱类似物以获得具有低毒性和高溶解度的化合物。目前,有两种药物用于临床实践,即CPT-11和托泊替康。其他衍生物,诸如贝洛替康、卢比替康、依沙替康、吉马替康、培加替康、勒托替康、karenitecin、阿非替康、高喜树碱、二氟替康及许多其他的衍生物,正在进行临床试验。化合物CPT-11是10-羟基-7-乙基喜树碱(常称作SN-38)的高度易溶的前药,被批准用于多种实体肿瘤和腹水(结直肠、皮肤、胃、肺、宫颈、卵巢、非霍奇金淋巴瘤)的治疗。
托泊替康是可溶于生理溶液的化合物,对肺、胃、肝、卵巢、乳腺、前列腺、食管、直肠、软组织肉瘤、头和颈、胶质母细胞瘤、慢性和急性髓性白血病这些肿瘤是有活性的。但是托泊替康显示重要的副作用诸如嗜中性白血球减少症和血小板减少症。
勒托替康是更易溶的衍生物,对颈、卵巢、乳腺、结直肠和肺部微细胞瘤这些肿瘤具有活性。但是,勒托替康亦具有血液毒性。
卢比替康是口服使用的有效对抗胰、卵巢和乳腺肿瘤的前药。
喜树碱及其类似物,作为完全拓扑异构酶I抑制剂,对常规药物、包括拓扑异构酶II抑制剂有耐药性的肿瘤有效;在整个细胞周期期间保持高的拓扑异构酶水平;未诱导多药耐药性(Pgo或MRP)或由酶介导的脱毒代谢。
目前,研究集中于具有比目前使用的药物更低毒性的拓扑异构酶I的新抑制剂。
开环喜树碱衍生物显示出高的蛋白结合(特别是与白蛋白)和低的肿瘤组织中的分布。结果,产物在身体中蓄积,对肿瘤的作用不佳。
相反地,内酯形式的高亲脂性促进喜树碱衍生物与细胞膜的吸附,特别是红细胞,影响了组织/血浆分布比。因此,目前研究集中于两种可选择的方法:a)设计仍具有好的溶解度的低蛋白结合产品;b)设计在极低剂量下具有治疗作用的高效产品。
在7、9、10和11位的修饰通常证实可被良好耐受,同时不影响DNA-拓扑异构酶I-喜树碱三元复合物的稳定性,其形成产生所述化合物的抗肿瘤活性。
具有20R构象的产物据证实与具有20S构象-与天然构象一致的产物相比无活性或活性很低。
通常,认为5位的修饰不利于三元复合物的生成,但是,已经报道吡啶酮环D和E的修饰对产物的活性不利。
发明内容
在第一方面,本发明涉及通式I的喜树碱衍生物、其可药用盐、异构体、对映体、非对映异构体和相应的混合物:
其中:
R为F、Cl、Br、I、-N3、NH2、-NR′R″、-COOR′、-CONR′R″、-NHR″′-NR′R″,其中R′、R″和R″′可为H、烷基、芳基、芳基烷基、酰基、烷氧基羰基、芳氧基羰基;
R1为烷基、氨基烷基、羟基烷基、腈、烷氧基亚氨基(alkoxymino)、芳氧基亚氨基(aryloxymino)、甲硅烷基烷基;
R2为氢、羟基、烷氧基、氨基烷基;
R3为氢,任选保护的羟基、烷氧基、氨基烷基;
其中直链或支链的烷基、酰基、烷氧基、氨基烷基或烷氧基亚氨基基团可包含1至8个,优选1至4个碳原子,且芳基和芳氧基基团可包含5至10个碳原子。
本发明的化合物显示低蛋白结合并具有良好的溶解度和在非常低的剂量下的高的药效。
用于制备本发明的化合物的优选的合成途径示于方案中并包含以下步骤:
a)前体羟基基团的保护;
b)通过碳负离子生成并与亲电试剂反应在5位衍生化;
c)羟基基团的脱保护;
方案
在方案中,R、R1、R2和R3具有上文所述的含义,且PG为羟基保护基团。
前体可以是可商购的或如文献所述地得到。
5位的碳负离子可通过用强有机碱、优选LiHMDS处理前体获得。
碳负离子原位与亲电试剂反应,亲电试剂诸如来源于卤素或氮杂二甲酸酯、异氰酸酯、氯羰基衍生物、甲苯磺酰基叠氮化物。
甲硅烷基和氨基甲酸酯或其组合优选为羟基保护性基团。
对本发明的化合物进行广谱肿瘤细胞的细胞毒性测定。以实施例的方式,报道了涉及两个式(I)化合物对NCI-H460细胞系(NSCL癌)的细胞毒性数据,使用喜树碱及药物托泊替康和SN-38作为参比标准:
(继续)
在DNA裂解测定中测定活性浓度和损伤持续性评价最有活性的化合物(见“实施例”节)。式(I)的衍生物在阻断DNA复制中与参比标准(特别地为托泊替康和喜树碱)相比令人惊异地显示较高的持续性,同时保持有效的细胞毒活性。
另一方面,本发明涉及包含式(I)化合物连同药物可接受的载体与赋形剂的药物组合物。适合口服或肠胃外施用的式(I)化合物的药物形式可以是固体,优选为胶囊、片剂和颗粒,或液体,优选为可注射的或输液溶液。
适合配成制剂的本发明的化合物可用于实体瘤和白血病的治疗,特别是肺、卵巢、乳腺、胃、肝、前列腺、软组织肉瘤、头和颈、食管、胰、结肠、直肠、胶质母细胞瘤、慢性和急性髓细胞白血病的肿瘤。
实施例
实施例I-20-OTES-喜树碱
将喜树碱(0.100g,0.287mmol)在惰性气氛下混悬于无水二甲基甲酰胺(3mL)中,得到的混悬液加以咪唑(0.980g,1.44mmol)。将混合物搅拌10分钟,接着向其中滴入氯化三乙基甲硅烷(TES-Cl)(0.193mL,1.15mmol),接着加入4-二甲基氨基吡啶(DMAP)(0.040g,0.287mmol)。46h后,真空蒸发反应混合物,(TLC控制试剂的完全消失,展开剂为CH2Cl2/MeOH=30/1)。随后将固体再溶于CH2Cl2中并用H2O和饱和NH4Cl洗涤。水相用CH2Cl2(2×10mL)萃取。合并有机相并用Na2SO4干燥,过滤并真空浓缩,由此得到淡黄色固体状的所需产物(0.133g,0.287mmol)。
1H NMR(CDCl3,400MHz)δ8.37(s,1H,Ar,H-7),8.25(d,1H,J=8.4Hz,Ar),7.92(d,1H,J=8.0Hz,Ar),7.82(t,1H,J=8.0Hz,Ar),7.65(t,1H,J=8.4Hz,Ar),7.57(s,1H,H-14),5.67(d,1H,J=16.4Hz,H-17),5.29(s,2H,H-5),5.25(d,1H,J=16.4Hz,H-17),2.00-1.84(m,2H,H-19),1.03-0.93(m,12H),0.80-0.71(m,6H)。13C NMR(CDCl3,100MHz)δ171.7,157.6,152.5,151.5,149.0,145.9,130.9,130.4,130.0,128.4,128.1,128.0,127.9,118.9,94.4,75.3,66.0,50.0,33.2,7.9,7.2,6.4。
实施例II-20-OTES SN-38
将SN-38(0.100g,0.255mmol)在惰性气氛下混悬于无水二甲基甲酰胺(5mL)中,得到的混悬液加以咪唑(0.087g,1.28mmol)。将混合物搅拌10分钟,接着向其中滴入氯化三乙基甲硅烷(TES-Cl)(0.171mL,1.02mmol),接着加入4-二甲基氨基吡啶(DMAP)(0.031g,0.255mmol)。52h后,真空蒸发反应混合物,用TLC监控(CH2Cl2/MeOH=10/1)试剂的完全消失。随后将固体再溶于CH2Cl2中并用H2O和饱和NH4Cl洗涤。水相用CH2Cl2(2×10mL)萃取。合并有机相并用Na2SO4干燥,过滤并真空浓缩,由此得到淡黄色固体状的所需产物(0.121g,0.240mmol,94%)。
1H NMR(CDCl3,400MHz)δ9.26(br s,1H,OH),8.14(d,1H,J=9.2Hz,Ar,H-12),7.58(s,1H,H-14),7.49(dd,1H,J1=9.2Hz J2=2.2Hz,H-11),7.46(d,1H,J=2.2Hz,H-9),5.70(d,1H,J=16.5Hz,H-17),5.28(d,1H,J=16.5Hz,H-17),5.23(s,2H,H-5),3.05(q,2H,J=7.5Hz),1.97-1.81(m,2H,H-19),1.32(t,3H,J=7.5Hz,Me),0.98-0.88(m,12H),0.77-0.68(m,6H)。13C NMR(CDCl3,100MHz)δ172.1,157.9,156.6,152.1,149.0,146.7,144.6,143.6,131.9,128.7,126.9,122.8,117.9,105.5,98.5,75.4,65.9,49.5,32.9,23.2,13.5,7.8,7.2,6.4。
实施例III-10-OTBDMS-20-OTES SN-38
将20-OTES SN-38(0.121g,0.240mmol)在惰性气氛下溶解于CH2Cl2/THF=1∶1(8mL)的无水混合物中。接着向其中加入咪唑(0.081g,1.20mmol),10分钟后加入叔丁基二甲基氯化甲硅烷(TBDMS-Cl)(0.144mg,0.957mmol),然后加入4-二甲基氨基吡啶(DMAP)(0.029g,0.240mmol)。18h后,真空蒸发反应混合物,TLC监控(己烷/AcOEt=1/1)试剂的消失。然后将固体再溶于CH2Cl2中并用H2O和饱和NH4Cl洗涤。水相用CH2Cl2(2×10mL)萃取,合并有机相,用Na2SO4干燥,过滤并真空浓缩。残留物用闪式色谱(SiO2,己烷/AcOEt=1/1)纯化,由此得到淡黄色固体状的所需产物(0.127g,0.205mmol,85%)。
1H NMR(CDCl3,400MHz)δ8.14(d,1H,J=8.8Hz,Ar,H-12),7.49(s,1H,H-14),7.40(d,1H,J=2.2Hz,H-9),7.38(dd,1H,J1=8.8Hz J2=2.5Hz,H-11),5.67(d,1H,J=16.5Hz,H-17),5.25(d,1H,J=16.5Hz,H-17),5.23(s,2H,H-5),3.11(q,2H,J=7.6Hz),1.99-1.82(m,2H,H-19),1.38(t,3H,J=7.6Hz,Me),1.04(s,9H),1.00-0.92(m,12H),0.78-0.69(m,6H),0.30(s,6H)。13C NMR(CDCl3,100MHz)δ171.9,157.7,155.1,151.5,150.1,146.8,145.6,143.5,132.2,128.2,126.9,125.9,118.0,110.5,97.7,75.4,66.0,49.3,33.2,25.6,23.1,18.3,13.7,7.9,7.2,6.4,4.3。
实施例IV-20-OTES托泊替康
将托泊替康(0.100g,0.238mmol)在惰性气氛下混悬于无水二甲基甲酰胺(5mL)中,得到的混悬液加以咪唑(0.081g,1.19mmol)。将混合物搅拌10分钟,接着向其中滴入氯化三乙基甲硅烷(TES-Cl)(0.160mL,0.952mmol),接着加入4-二甲基氨基吡啶(DMAP)(0.029g,0.238mmol)。52h后,真空蒸发反应混合物,用TLC监控(CH2Cl2/MeOH=10/1)试剂的完全消失。随后将固体再溶于CHCl3、H2O和饱和NH4Cl中,水相用CHCl3(2×15mL)萃取。合并有机相并用Na2SO4干燥,过滤并真空浓缩,由此得到淡黄色固体状的所需产物(0.120g,0.224mmol,94%)。
1H NMR(CDCl3,400MHz)δ9.65(br s,1H),8.26(s,1H,Ar,H-7),8.14(d,1H,J=8.8Hz,Ar,H-12),7.80(d,1H,J=8.8Hz,Ar,H-11),7.58(s,1H,H-14),5.67(d,1H,J=16.5Hz,H-17),5.25(d,1H,J=16.5Hz,H-17),5.20(s,2H,H-5),4.71(s,2H),2.81(s,6H,2Me),1.97-1.81(m,2H,H-19),0.98-0.88(m,12H),0.77-0.68(m,6H)。13C NMR(CDCl3,100MHz)δ172.1,157.9,156.6,152.1,150.8,146.8,144.3,134.3,131.2,129.9,127.9,123.0.118.9,110.1,98.5,75.4,65.9,51.1,50.0.43.1,32.9,7.8,7.2,6.4。
实施例V-10-OTBDMS 20-OTES托泊替康
将20-OTES托泊替康(0.120g,0.224mmol)在惰性气氛下溶解于CH2Cl2/THF=1∶1无水混合物(8mL)中。接着加入咪唑(0.076g,1,12mmol),10分钟后,加入叔丁基二甲基氯化甲硅烷(TBDMS-Cl)(0.135mg,0.896mmol),接着加入4-二甲基氨基吡啶(DMAP)(0.027g,0.224mmol)。21h后,真空蒸发反应混合物,用TLC(己烷/AcOEt=1/1)监控试剂的消失。然后将固体再溶于CHCl3、H2O和饱和NH4Cl中,水相用CHCl3(2×15mL)萃取。合并有机相并用Na2SO4干燥,过滤并真空浓缩。残留物用闪式色谱(SiO2,己烷/AcOEt=1/1)纯化,由此得到淡黄色固体状的所需产物(0.116g,0.179mmol,80%)。
1H NMR(CDCl3,400MHz)δ8.26(s,1H,Ar,H-7),8.14(d,1H,J=8.8Hz,Ar,H-12),7.81(d,1H,J=8.8Hz,Ar,H-11),7.59(s,1H,H-14),5.64(d,1H,J=16.5Hz,H-17),5.22(d,1H,J=16.5Hz,H-17),5.19(s,2H,H-5),4.71(s,2H),2.81(s,6H,2Me),1.97-1.81(m,2H,H-19),1.04(s,9H),0.98-0.88(m,12H),0.77-0.68(m,6H),0.30(s,6H)。13C NMR(CDCl3,100MHz)δ171.7,157.7,155.1,151.5,150.0.146.8,144.3,134.3,131.2 129.9,127.9,123.0,118.9,110.1,98.5,75.4,65.9,51.1,50.0.43.9,32.9,25.6,18.3,7.8,7.2,6.4,-4.3。
实施例VI-20-OTES 10-羟基喜树碱的制备
将10-羟基喜树碱(0.100g,0.275mmol)在惰性气氛下混悬于无水二甲基甲酰胺(5mL)中,得到的混悬液中加入咪唑(0.225g,3.31mmol)。将混合物搅拌10分钟,接着向其中滴入三乙基氯化甲硅烷(TES-Cl)(0.460mL,2.75mmol),接着加入4-二甲基氨基吡啶(DMAP)(0.068g,0.550mmol)。24h后,真空蒸发反应混合物,用TLC(CH2Cl2/MeOH=20/1)监控试剂的完全消失。随后将固体再溶于CH2Cl2中,并用H2O和饱和NH4Cl洗涤。水相用CH2Cl2(2×10mL)萃取。合并有机相并用Na2SO4干燥,过滤并真空浓缩,由此得到淡黄色固体状的所需产物(0.124g,0.259mmol,94%)。
1H NMR(CDCl3+5%CD3OD,400MHz)δ8.10(s,1H,Ar,H-7),8.05(d,1H,J=9.2Hz,Ar),7.50(s,1H,H-14),7.39(dd,1H,J1=9.2HzJ2=2.4Hz,H-11),7.11(d,1H,J=2.2Hz,H-9),5.60(d,1H,J=16.4Hz,H-17),5.21(d,1H,J=16.4Hz,H-17),5.15(s,2H,H-5),1.97-1.81(m,2H,H-19),0.98-0.88(m,12H),0.76-0.68(m,6H)。13C NMR(CDCl3+5%CD3OD,100MHz)δ172.2,157.8,156.7,151.8,149.2,146.1,144.1,130.9,129.8,129.0,128.6,123.2,117.8,108.8,98.1,75.4,65.8,50.0,32.9,7.7,7.1,6.3。
实施例VII-10-OTBDMS-20-OTES喜树碱
将10-羟基-20-OTES-喜树碱(0.105g,0.219mmol)在惰性气氛下溶解于CH2Cl2/THF=1∶1无水混合物(4mL)中。接着加入咪唑(0.097g,1.42mmol),10分钟后,加入叔丁基二甲基氯化甲硅烷(TBDMS-Cl)(0.164mg,1.10mmol),接着加入4-二甲基氨基吡啶(DMAP)(0.040g,0.329mmol)。18h后,真空蒸发反应混合物,用TLC监控(环己烷/AcOEt=1/3)试剂的完全消失。随后将固体再溶于CH2Cl2中,并用H2O和饱和NH4Cl洗涤。水相用CH2Cl2(2×10mL)萃取。合并有机相并用Na2SO4干燥,过滤并真空浓缩。残留物用闪式色谱(SiO2,环己烷/AcOEt=1/3)纯化,由此得到淡黄色固体状的所需产物(0.117g,0.197mmol,90%)。
1H NMR(CDCl3,400MHz)δ8.22(s,1H,Ar,H-7),8.13(d,1H,J=9.2Hz,Ar,H-12),7.51(s,1H,H-14),7.39(dd,1H,J1=9.2Hz J2=2.8Hz,H-11),7.22(d,1H,J=2.8Hz,H-9),5.66(d,1H,J=16.5Hz,H-17),5.25(s,2H,H-5),5.24(d,1H,J=16.5Hz,H-17),1.99-1.82(m,2H,H-19),1.03(s,9H),1.00-0.92(m,12H),0.78-0.69(m,6H),0.29(s,6H)。13C NMR(CDCl3,100MHz)δ172.0,157.7,155.1,151.5,150.6,146.1,145.1,131.4,129.4,129.3,128.7,126.7,118.3,114.5,97.7,75.3,66.0,49.9,33.1,25.6,18.3,7.9,7.2,6.4,-4.3。
实施例VIII-5-F-20-OTES-喜树碱
将喜树碱20-OTES(0.100g,0.216mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,然后冷却至-78℃,并向其中滴入1.0M LiHMDS在THF(0.260mL,0.260mmol)中的溶液。20分钟后,加入无水THF(2mL)中的NFSI(0.089g,0.281mmol)。-78℃2h后,使温度升高至25℃,并通过TLC(己烷/AcOEt=1/2)监控试剂的消失。观察到两种非对映异构体的生成。室温下3h后,加入饱和NH4Cl终止反应。水相用CH2Cl2(3×15mL)萃取,并合并有机相,用Na2SO4干燥,过滤并真空浓缩。残留物用闪式色谱纯化(SiO2,环己烷/AcOEt=3/1,然后2/1,最终1/1),由此得到淡黄色固体状的两种异构体的混合物(0.101g,0.210mmol,97%)(异构体比例为1∶1)。通过进一步的色谱处理分离两种异构体。洗脱顺序如下:
第一种非对映异构体:1H NMR(CDCl3,400MHz)δ8.52(s,1H,Ar,H-7),8.25(d,1H,J=8.4Hz,Ar),7.96(d,1H,J=8.4Hz,Ar),7.87(t,1H,J=8.4Hz,Ar),7.69(t,1H,J=8.4Hz,Ar),7.47(d,1H,1JHF=61.2Hz,H-5),7.45(s,1H,H-14),5.62(d,1H,J=16.8Hz,H-17),5.22(d,1H,J=16.8Hz,H-17),2.02-1.84(m,2H,H-19),1.03-0.93(m,12H),0.80-0.71(m,6H)。13C NMR(CDCl3,100MHz)δ171.4,157.5,152.3,151.1,150.2(d,J=1.5Hz),150.3(d,J=1.5Hz),143.6(d,J=5.3Hz),133.7,131.7,130.2,128.9,128.4,127.9(d,J=15.0Hz),126.3(d,J=15.0Hz),121.8,98.9,93.8(d,1JCF=213.2Hz,C-5),75.1,65.7,33.1,7.8,7.2,6.4。
第二种非对映异构体:1H NMR(CDCl3,400MHz)δ8.51(s,1H,Ar,H-7),8.25(d,1H,J=8.4Hz,Ar),7.96(d,1H,J=8.4Hz,Ar),7.87(t,1H,J=8.4Hz,Ar),7.68(t,1H,J=8.4Hz,Ar),7.51(d,1H,1JHF=60.8Hz,H-5),7.42(s,1H,H-14),5.62(d,1H,J=17.2Hz,H-17),5.20(d,1H,J=17.2Hz,H-17),2.02-1.82(m,2H,H-19),1.04-0.93(m,12H),0.80-0.71(m,6H)。13C NMR(CDCl3,100MHz)δ171.2,157.8,152.5,151.2,150.3,143.7,133.7(d,J=2.4Hz),131.7,130.2,128.9,128.3,127.9(d,J=2.3Hz),126.3(d,J=16.7Hz),121.8(d,J=1.5Hz),99.0,93.8(d,1JCF=214.8Hz,C-5),75.0,65.8,33.3,7.9,7.1,6.4。
实施例IX-5-F-喜树碱第一种非对映异构体的制备
将5-F-20-OTES-喜树碱的第一种非对映异构体(0.025g,0.052mmol)在惰性气氛下搅拌溶解于无水THF(5mL)中。接着向其中滴入Et3N·3HF(0.060mL,0.368mmol)。将反应混合物室温下反应28h,用TLC监控试剂的消失(己烷/AcOEt=1/2)。真空下挥干溶剂,残留物进行色谱处理(SiO2,己烷/AcOEt=1/1),由此得到淡黄色固体状的所需产物(0.019g,0.051mmol,98%)。
1H NMR(CDCl3,400MHz)δ8.52(s,1H,Ar,H-7),8.25(d,1H,J=8.4Hz,Ar),7.96(d,1H,J=8.4Hz,Ar),7.87(t,1H,J=8.4Hz,Ar),7.69(t,1H,J=8.4Hz,Ar),7.59(s,1H,H-14),7.46(d,1H,1JHF=61.2Hz,H-5),5.69(d,1H,J=16.8Hz,H-17),5.26(d,1H,J=16.8Hz,H-17),3.87(br s,1H,OH),2.01-1.81(m,2H,H-19),1.05(t,3H,J=7.6Hz,Me)。13C NMR(CDCl3,100MHz)δ173.5,157.6,151.1,151.0.150.2,144.1,133.9,131.9,130.0,129.0,128.5,127.8,126.4,121.7,98.8,93.8(d,1JCF=214.0Hz,C-5),72.5,66.0,31.5,7.8。
实施例X-5-F-喜树碱第二种非对映异构体的制备
将5-F-20-OTES-喜树碱的第二种非对映异构体(0.025g,0.052mmol)在惰性气氛下搅拌溶解于无水THF(5mL)中,接着向其中滴入Et3N·3HF(0.060mL,0.368mmol)。将反应混合物室温下反应28h,用TLC监控试剂的消失(己烷/AcOEt=1/2)。真空下挥干溶剂,残留物进行色谱处理(SiO2,己烷/AcOEt=1/1),由此得到淡黄色固体状的所需产物(0.018g,0.050mmol,97%)。
1H NMR(CDCl3,400MHz)δ8.52(s,1H,Ar,H-7),8.24(d,1H,J=8.4Hz,Ar),7.96(d,1H,J=8.4Hz,Ar),7.88(t,1H,J=8.4Hz,Ar),7.69(t,1H,J=8.4Hz,Ar),7.56(s,1H,H-14),7.51(d,1H,1JHF=60.4Hz,H-5),5.69(d,1H,J=16.4Hz,H-17),5.25(d,1H,J=16.4Hz,H-17),3.87(br s,1H,OH),1.98-1.78(m,2H,H-19),1.04(t,3H,J=7.6Hz,Me)。13C NMR(CDCl3,100MHz)δ173.3,157.7,151.2,151.2,150.2,144.2,133.8,131.9,130.0,129.0,128.5,127.8,126.4,121.6,98.9,93.7(d,1JCF=214.0Hz,C-5),72.5,66.1,31.6,7.8。
实施例XI-5-N3-20-OTES-喜树碱的制备
将喜树碱20-OTES(0.100g,0.216mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,然后冷却至-78℃,并向其中滴入1.0M LiHMDS在THF(0.260mL,0.260mmol)中的溶液。20分钟后,加入无水THF(2mL)中的甲苯磺酰基叠氮化物(TsN3)(0.055g,0.281mmol)。在-78℃下2h后,使温度升高至25℃,并通过TLC监控试剂的消失(己烷/AcOEt=2/1)。观察到两种非对映异构体的生成。室温下2h 30min后,加入饱和NH4Cl终止反应。水相用CH2Cl2(3×15mL)萃取,并合并有机相,用Na2SO4干燥,过滤并真空浓缩。由两种非对映异构体组成的残留物用闪式色谱(SiO2,环己烷/AcOEt=3/1,然后2/1,最终1/1)纯化,由此得到淡黄色固体状的两种异构体的混合物(0.106g,0.210mmol,97%)(异构体比例为1∶1)。通过进一步的色谱处理分离两种异构体。洗脱顺序如下:
第一种非对映异构体:1H NMR(CDCl3,400MHz)δ8.45(s,1H,Ar,H-7),8.25(d,1H,J=8.4Hz,Ar),7.95(d,1H,J=8.4Hz,Ar),7.86(t,1H,J=8.4Hz,Ar),7.68(t,1H,J=8.4Hz,Ar),7.49(s,1H,H-14),6.97(s,1H,H-5),5.65(d,1H,J=16.8Hz,H-17),5.26(d,1H,J=16.8Hz,H-17),2.01-1.84(m,2H,H-19),1.03-0.94(m,12H),0.80-0.71(m,6H)。13C NMR(CDCl3,100MHz)δ171.6,158.3,152.2,150.8,150.0,144.0,132.9,131.4,130.1,128.6,128.3,128.2,128.1,120.8,98.7,75.4,75.2,65.7,33.1,7.9,7.2,6.4。
第二种非对映异构体:1H NMR(CDCl3,400MHz)δ8.45(s,1H,Ar,H-7),8.24(d,1H,J=8.4Hz,Ar),7.95(d,1H,J=8.4Hz,Ar),7.86(t,1H,J=8.4Hz,Ar),7.68(t,1H,J=8.4Hz,Ar),7.46(s,1H,H-14),6.99(s,1H,H-5),5.66(d,1H,J=16.8Hz,H-17),5.22(d,1H,J=16.8Hz,H-17),2.02-1.84(m,2H,H-19),1.03-0.94(m,12H),0.80-0.71(m,6H)。13C NMR(CDCl3,100MHz)δ171.4,158.4,152.3,150.9,150.0,144.0,132.9,131.4,130.1,128.6,128.3,128.2,128.1,120.8,98.7,75.3,75.1,65.8,33.3,7.9,7.2,6.4。
实施例XII-5-N3-喜树碱第一种非对映异构体的制备
将5-N3-20-OTES-喜树碱的非对映异构体1(0.070g,0.139mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,接着向其中滴入Et3N·3HF(0.170mL,1.016mmol)。将反应混合物室温下反应26h,用TLC(己烷/AcOEt=1/1)监控试剂的消失。真空下挥干溶剂,残留物用闪式色谱(SiO2,己烷/AcOEt=1/1)纯化,由此得到淡黄色固体状的所需产物(0.053g,0.136mmol,98%)。
1H NMR(CDCl3,400MHz)δ8.44(s,1H,Ar,H-7),8.24(d,1H,J=8.4Hz,Ar),7.93(d,1H,J=8.4Hz,Ar),7.85(t,1H,J=8.4Hz,Ar),7.67(t,1H,J=8.4Hz,Ar),7.63(s,1H,H-14),6.97(s,1H,H-5),5.70(d,1H,J=16.8Hz,H-17),5.29(d,1H,J=16.8Hz,H-17),3.99(br s,1H,OH),2.00-1.84(m,2H,H-19),1.04(t,3H,J=7.6Hz,Me)。13C NMR(CDCl3,100MHz)δ173.6,158.3,150.8,150.7,149.8,144.4,133.1,131.5,129.9,128.6,128.3,128.3,128.1,120.6,98.6,75.4,72.7,66.0,31.5,7.8。
实施例XIII-5-N3-喜树碱第二种非对映异构体的制备
5-N3-20-OTES-喜树碱的非对映异构体2(0.055g,0.109mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,接着向其中滴入Et3N·3HF(0.135mL,0.820mmol)。反应混合物室温下反应26h,用TLC监控起始试剂的消失(己烷/AcOEt=1/1)。真空下挥干溶剂,残留物通过闪式色谱(SiO2,己烷/AcOEt=1/1)纯化,由此得到淡黄色固体状的所需产物(0.042g,0.107mmol,98%)。
1H NMR(CDCl3,400MHz)δ8.45(s,1H,Ar,H-7),8.23(d,1H,J=8.4Hz,Ar),7.95(d,1H,J=8.4Hz,Ar),7.85(t,1H,J=8.4Hz,Ar),7.68(t,1H,J=8.4Hz,Ar),7.60(s,1H,H-14),7.00(s,1H,H-5),5.74(d,1H,J=16.8Hz,H-17),5.28(d,1H,J=16.8Hz,H-17),3.86(br s,1H,OH),1.98-1.82(m,2H,H-19),1.04(t,3H,J=7.6Hz,Me)。13C NMR(CDCl3,100MHz)δ173.4,158.4,150.9,150.7,149.8,144.5,133.0,131.5,129.9,128.6,128.4,128.3,128.1,120.6,98.6,75.3,72.6,66.1,31.6,7.8。
实施例XIV-5-NH2-喜树碱的制备
将5-N3-20-OH-喜树碱的非对映异构体2(0.050g,0.129mmol)在惰性气氛下搅拌溶解于无水THF(1.5mL)和无水MeOH(6mL)的混合物中,接着加入Pd/C(14mg~10%),进行两个周期的真空/H2(H2气球压力)。将反应混合物室温下反应3h,用TLC(己烷/AcOEt=1/3)监控试剂的消失,然后通过硅藻土过滤并用CH2Cl2(2×15mL)洗涤。真空下挥干溶剂。反应粗品的1H NMR谱显示所需产物以C5位上两种差向异构体的1∶1混合物存在。闪式色谱(SiO2,CH2Cl2/MeOH=35/1然后25/1)可回收两种非对映异构体的混合物(0.046g,0.126mmol,98%)。
1H NMR(CDCl3,400MHz)δ8.48(s,1H,Ar,H-7),8.22-8.17(m,1H,Ar),7.95-7.90(m,1H,Ar),7.85-7.78(m,1H,Ar),7.68-7.60(m,1H,Ar),7.58(s,0.5H,H-14),7.54(s,0.5H,H-14)6.50(s,0.5H,H-5),6.47(s,0.5H,H-5),5.74-5.64(m,1H,H-17),5.28-5.22(m,1H,H-17),4.00-2.40(br s,3H,OH+NH2),1.98-1.82(m,2H,H-19),1.07-1.01(m,3H,Me)。13C NMR(CDCl3,100MHz)δ173.8(2C),158.5(2C),151.2(2C),150.4(2C),149.7(2C),144.5(2C),132.7(2C),131.0(2C),129.8(2C),128.5(2C),128.3(2C),128.0(2C),127.8(2C),120.2(2C),113.8(2C),97.7(2C),72.7(2C),66.3,66.0,31.5(2C),7.8,7.8。
实施例XV-5-二-叔-丁氧基羰基肼基-20-OTES-喜树碱
将喜树碱20-OTES(0.100g,0.216mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,然后冷却至-78℃,并向其中滴入1.0M LiHMDS在THF(0.281mL,0.281mmol)中的溶液。20分钟后,加入在无水THF(2mL)中的偶氮二甲酸二叔丁酯(DTBAC)(0.075g,0.324mmol)。-78℃下4h后,用TLC(己烷/AcOEt=3/1)监控试剂的消失。观察到生成了两种非对映异构体。加入饱和NH4Cl终止反应。水相用CH2Cl2(3×15mL)萃取,并合并有机相,用Na2SO4干燥,过滤并真空浓缩。残留物用闪式色谱纯化(SiO2,环己烷/AcOEt=3/1),由此得到两种异构体的混合物(0.145g,0.210mmol,97%)。通过进一步的色谱处理分离两种异构体。洗脱顺序如下:
第一种非对映异构体:1H NMR(CDCl3,400MHz)δ8.80(br s,1H,Ar),8.23(d,1H,J=8.4Hz,Ar),8.01(br d,1H,Ar),7.90-7.71(m,2H,Ar),7.70-7.45(m,2H,Ar+H-14),6.52(br s,1H,H-5),5.61(d,1H,J=16.8Hz,H-17),5.23(d,1H,J=16.8Hz,H-17),2.03-1.81(m,2H,H-19),1.79-1.08(br s,18H),1.06-0.92(m,12H),0.80-0.70(m,6H)。13C NMR(CDCl3,100MHz)δ171.7,157.8,155.5,155.5,152.0,152.0,151.2,149.4,145.0,132.1,130.6,130.0,128.7,128.4,127.9,119.9,98.2,82.7,81.5,79.7,75.2,65.7,33.2,28.3,27.6,7.7,7.2,6.4。
第二种非对映异构体:1H NMR(CDCl3,400MHz)δ8.79(br s,1H,Ar),8.23(d,1H,J=8.4Hz,Ar),8.01(br d,1H,Ar),7.85-7.76(m,2H,Ar),7.65(br t,1H,J=8.4Hz,Ar),7.52(s,1H,H-14),6.54(br s,1H,H-5),5.61(d,1H,J=16.8Hz,H-17),5.22(d,1H,J=16.8Hz,H-17),2.03-1.82(m,2H,H-19),1.76-1.08(br s,18H),1.04-0.92(m,12H),0.80-0.70(m,6H)。13C NMR(CDCl3,100MHz)δ171.5,157.9,155.5,155.5,152.3,152.0.151.2,149.4,145.1,132.1,130.6,130.0,128.7,128.4,127.9,119.9,98.2,82.9,81.5,79.6,75.2,65.8,33.3,28.3,27.4,7.8,7.2,6.4。
实施例XVI-5-二-叔-丁氧基羰基肼基-喜树碱第一种非对映异构体的制备
将5-二-叔-丁氧基羰基肼基-20-OTES-喜树碱(0.050g,0.072mmol)第一种非对映异构体在惰性气氛下搅拌溶解于无水THF(4mL)中,接着向其中滴入Et3N·3HF(0.088mL,0.542mmol)。反应混合物室温下反应35h,用TLC监控试剂的消失(己烷/AcOEt=3/2)。真空下挥干溶剂,残留物用闪式色谱(SiO2,己烷/AcOEt=3/2)纯化,由此得到淡黄色固体状的所需化合物(0.041g,0.071mmol,98%)。
产物通过CH2Cl2/戊烷=1/50结晶进一步纯化。
1H NMR(CDCl3,400MHz)δ8.77(br s,1H,Ar),8.16(br d,1H,J=8.0Hz,Ar),7.97(br s,1H,Ar),7.86-7.50(m,4H,Ar),6.51(br s,1H,H-5),5.66(d,1H,J=16.4Hz,H-17),5.24(d,1H,J=16.4Hz,H-17),3.86(br s,1H,OH),2.00-1.80(m,2H,H-19),1.79-1.13(br s,18H),1.03(t,3H,J=7.6Hz,Me)。13C NMR(CDCl3,100MHz)δ173.7,157.9,155.5,155.5,152.1,151.3,150.7,149.6,145.7,132.3,130.7,129.9,128.7,127.9,127.6,120.0,97.9,82.8,81.6,79.7,72.7,66.1,31.8,28.3,27.7,7.7。
实施例XVII-5-二-叔-丁氧基羰基肼基-喜树碱第二种非对映异构体的制备
5-二-叔-丁氧基羰基肼基-20-OTES-喜树碱(0.050g,0.072mmol)第二种非对映异构体在惰性气氛下搅拌溶解于无水THF(4.5mL)中,接着向其中滴入Et3N·3HF(0.088mL,0.542mmol)。反应混合物室温下反应35h,用TLC监控试剂的消失(己烷/AcOEt=3/2)。真空下挥干溶剂,残留物用闪式色谱(SiO2,己烷/AcOEt=3/2)纯化,由此得到淡黄色固体状的所需化合物(0.040g,0.069mmol,96%)。产物通过CH2Cl2/戊烷=1/50结晶进一步纯化。
1H NMR(CDCl3,400MHz)δ8.79(br s,1H,Ar),8.22(br d,1H,J=8.4Hz,Ar),7.99(br s,1H,Ar),7.88-7.50(m,4H,Ar),6.53(br s,1H,H-5),5.65(d,1H,J=16.4Hz,H-17),5.26(d,1H,J=16.4Hz,H-17),3.80(br s,1H,OH),2.00-1.80(m,2H,H-19),1.79-1.13(br s,18H),1.03(t,3H,J=7.2Hz,Me)。13C NMR(CDCl3,100MHz)δ173.6,157.9,155.4,155.4,152.1,151.3,150.8,149.5,145.6,132.3,130.8,129.8,128.7,127.9,127.8,119.8,98.0,83.0,81.5,79.7,72.7,66.3,31.8,28.3,27.7,7.8。
实施例XVIII-5-二苄基氧基羰基肼基-20-OTES-喜树碱的制备
将喜树碱20-OTES(0.100g,0.216mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,然后冷却至-78℃,并向其中滴入1.0M LiHMDS在THF(0.281mL,0.281mmol)中的溶液。20分钟后,向其中加入在无水THF(2mL)中的偶氮二甲酸二苄酯(0.097g,0.324mmol)。-78℃下3h后,使温度升高至25℃并用TLC(己烷/AcOEt=3/1)监控试剂的消失。观察到生成了两种非对映异构体。室温90分钟后,加入饱和NH4Cl终止反应。水相用CH2Cl2(3×15mL)萃取,并合并有机相,用Na2SO4干燥,过滤并真空浓缩。残留物用闪式色谱(SiO2,环己烷/AcOEt=4/1然后7/2)纯化,由此得到淡黄色固体(0.161g,0.212mmol,98%)。通过进一步的色谱处理分离两种异构体。洗脱顺序如下:
第一种非对映异构体:1H NMR(CDCl3,400MHz)δ8.70(br s,1H,Ar),8.39(br s 1H,Ar),8.22(br d,1H,J=7.6Hz,Ar),7.95(br d,1H,J=7.6Hz,Ar),7.83(br t,1H,J=7.6Hz,Ar),7.65(br t,1H,J=7.6Hz,Ar),7.64-7.00(m,11H,Ar+H-14),6.49(br s,1H,H-5),5.57(d,1H,J=16.4Hz,H-17),5.47-4.44(m,5H),1.98-1.82(m,2H,H-19),1.02-0.89(m,12H),0.80-0.70(m,6H)。13C NMR(CDCl3,100MHz)δ171.6,158.0.156.3,156.3,153.0,152.2,151.0,149.6,144.8,135.3,132.1,130.6,130.0,128.6-127.8(11C),119.9,98.4,79.5,75.2,68.4,67.9,65.6,33.0,7.9,7.2,6.4。
第二种非对映异构体:1H NMR(CDCl3,400MHz)δ8.85(br s,1H,Ar),8.58(br s 1H,Ar),8.20(br s,1H,Ar),7.93(br s,Ar),7.81(br t,1H,J=7.6Hz,Ar),7.63(br t,1H,J=7.6Hz,Ar),7.56-6.90(m,11H,Ar+H-14),6.52(br s,1H,H-5),5.55(d,1H,J=16.8Hz,H-17),5.44-4.71(m,5H),1.98-1.80(m,2H,H-19),1.05-0.90(m,12H),0.81-0.70(m,6H)。13C NMR(CDCl3,100MHz)δ171.5,157.9,156.4,156.4,152.9,152.4,150.9,149.4,144.8,135.3,132.1,130.6,129.9,128.6-127.8(11C),119.9,98.5,79.3,75.2,68.4,67.8,65.6,32.9,7.8,7.2,6.4。
实施例XIX-5-二苄基氧基羰基肼基-喜树碱第一种非对映异构体的制备
将5-二苄基氧基羰基肼基-20-OTES-喜树碱第一种非对映异构体(0.140g,0.184mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,接着向其中滴入Et3N·3HF(0.225mL,1.380mmol)。反应混合物室温下反应52h,用TLC监控试剂的消失(己烷/AcOEt=1/3)。真空下挥干溶剂,残留物用闪式色谱(SiO2,己烷/AcOEt=1/1,然后2/3)纯化,由此得到淡黄色固体(0.113g,0.175mmol,95%)。产物通过CH2Cl2/戊烷=1/50结晶进一步纯化。
1H NMR(CDCl3,400MHz)δ8.67(br s,1H,Ar),8.39(br s 1H,Ar),8.12(br d,1H,J=7.6Hz,Ar),7.95(br s,1H,Ar),7.74(br t,1H,J=7.6Hz,Ar),7.65-6.66(m,12H,Ar+H-14),6.48(br s,1H,H-5),5.55(d,1H,J=16.0Hz,H-17),5.42-4.44(m,5H),3.86(br s,1H,OH),1.92-1.72(m,2H,H-19),0.95(t,3H,J=7.6Hz,Me)。13C NMR(CDCl3,100MHz)δ173.5,158.0,156.2,156.0,153.0,150.9,150.9,149.5,145.3,135.4,132.2,130.7,129.8,128.7-127.8(11C),119.9,98.2,79.6,72.7,68.5,68.0,65.9,31.6,7.8。
实施例XX-5-二苄基氧基羰基肼基-喜树碱第二种非对映异构体的制备
将5-二苄基氧基羰基肼基-20-OTES-喜树碱的第二种非对映异构体(0.140g,0.184mmol)在惰性气氛下搅拌溶解于无水THF(6mL)中,接着向其中滴入Et3N·3HF(0.150mL,0.921mmol)。反应混合物室温下反应55h,用TLC(己烷/AcOEt=3/2)监控试剂的消失。真空下挥干溶剂,残留物用闪式色谱(SiO2,己烷/AcOEt=1/1)纯化,由此得到淡黄色固体状的所需化合物(0.113g,0.175mmol,95%)。产物通过CH2Cl2/戊烷=1/50结晶进一步纯化。
1H NMR(CDCl3,400MHz)δ8.71(br s,1H,Ar),8.34(br s 1H,Ar),8.18(br s,1H,Ar),7.94(br s,1H,Ar),7.79(br t,1H,J=7.6Hz,Ar),7.70-6.70(m,12H,Ar+H-14),6.52(br s,1H,H-5),5.53(d,1H,J=16.4Hz,H-17),5.44-4.48(m,5H),3.87(br s,1H,OH),1.90-1.70(m,2H,H-19),0.99(t,3H,J=7.6Hz,Me)。13C NMR(CDCl3,100MHz)δ173.4,158.0,156.3,156.1,153.0,151.0,150.9,149.6,145.3,135.5,132.3,130.8,129.8,128.7-127.8(11C),119.8,98.4,79.5,72.7,68.5,67.8,66.0.31.6,7.7。
实施例XXI-细胞生长抑制作用的测定
源于人体大细胞肺肿瘤的H460细胞在含10%胎牛血清的RPMI-1640培养基中培养。细胞敏感性通过在1或72hr药物暴露后的细胞生长抑制测定进行确定。收集处于对数生长的细胞并植入6-孔板中,试验双份。植板二十四小时后,将细胞暴露于药物,并在暴露于药物72小时后用库尔特计数器进行计数,用于确定IC50。IC50定义为与未处理的对照生长比较,细胞生长被抑制50%的浓度。
实施例XXII-拓扑异构酶-I-依赖的DNA破裂测定
DNA破裂使用751-bp BamHI-EcoRI DNA SV40纯化凝胶进行确定(Beretta GL,Binaschi M,Zagni AND,Capuani L,Capranico G.Tethering a type IB topoisomerase to a DNA site by enzyme fusion to aheterologous site-selective DNA-binding protein domain.Cancer Res1999;59:3689-97)。DNA碎片仅在3’标记。DNA破裂反应(20,000cpm/样本)在20ml的10mM Tris-HCL(pH 7.6)、150mM KCl、5mM MgCl2、15μg/mL BSA、0.1mM硫代苏糖醇和人重组酶(全长top1)中37℃下进行30min。反应使用0.5%SDS和0.3mg/mL蛋白酶K在42℃阻断45min。DNA损伤持续性在不同的时间加入0.6M NaCl与10μM的所述药物孵育30分钟后检验。沉淀后,DNA重混悬于变性缓冲液(80%甲酰胺,10mM NaOH,0.01M EDTA和1mg/mL染料)中,之后植入于变性凝胶(7%聚丙烯酰胺在TBE缓冲液中)。全部DNA的破碎水平用PhosphoImager 425型(分子动力学)进行测定(Dallavalle S,Ferrari A,Biasotti B,等人.Novel 7-oxyiminomethyl comptothecin derivatives withpotent in vitro and in vivo antitumor activity.J Med Chem 2001;44:3264-74)。
DNA损伤的持续(%)
Claims (10)
1.通式(I)的化合物、其可药用盐、非对映异构体和相应的混合物:
其中:
R为F;
R1为烷基、氨基烷基、羟基烷基、腈、甲硅烷基烷基;
R2为氢、羟基、烷氧基、氨基烷基;
R3为氢、羟基、烷氧基、氨基烷基;
其中直链或支链的烷基、烷氧基或氨基烷基基团包含1至8个碳原子。
2.化合物,其选自
a)5-F-喜树碱。
3.根据权利要求1的化合物,其中直链或支链的烷基、烷氧基或氨基烷基包含1至4个碳原子。
4.制备根据权利要求1的式(I)化合物或权利要求2的化合物的方法,包含下列方案中所示的步骤,其中:
a)前体羟基基团的保护;
b)通过碳负离子生成并与亲电试剂反应在5位衍生化;
c)羟基基团的脱保护;
方案
其中R、R1、R2和R3具有权利要求1或2所述的含义,且PG为羟基保护基团。
5.药物组合物,其含有根据权利要求1的式(I)的化合物或权利要求2的化合物和可药用的载体和赋形剂。
6.如权利要求5所述的药物组合物,其为适于口服和胃肠外施用的形式。
7.如权利要求1-3所述的化合物或如权利要求5-6所述的组合物用于制备治疗肿瘤的药物的用途。
8.权利要求7所述的用途,其中所述的药物用于治疗实体瘤和白血病。
9.权利要求8所述的用途,其中实体瘤为肺、卵巢、乳腺、胃、肝、前列腺、软组织肉瘤、食管、胰、头和颈、胶质母细胞瘤的肿瘤。
10.权利要求8所述的用途,其中白血病为慢性和急性髓细胞白血病。
Applications Claiming Priority (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
ITMI2006A001474 | 2006-07-26 | ||
IT001474A ITMI20061474A1 (it) | 2006-07-26 | 2006-07-26 | Derivati della camptotecina ad attivita antitumorale |
PCT/EP2007/006294 WO2008012003A1 (en) | 2006-07-26 | 2007-07-16 | Camptothecin derivatives with antitumor activity |
Publications (2)
Publication Number | Publication Date |
---|---|
CN101495485A CN101495485A (zh) | 2009-07-29 |
CN101495485B true CN101495485B (zh) | 2012-04-11 |
Family
ID=38668855
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN2007800281978A Active CN101495485B (zh) | 2006-07-26 | 2007-07-16 | 具有抗肿瘤活性的喜树碱衍生物 |
Country Status (20)
Country | Link |
---|---|
US (1) | US8193210B2 (zh) |
EP (1) | EP2044080B1 (zh) |
JP (1) | JP5313895B2 (zh) |
KR (1) | KR101496374B1 (zh) |
CN (1) | CN101495485B (zh) |
AU (1) | AU2007278509B2 (zh) |
BR (1) | BRPI0714672B8 (zh) |
CA (1) | CA2658902C (zh) |
DK (1) | DK2044080T3 (zh) |
ES (1) | ES2628022T3 (zh) |
HK (1) | HK1135692A1 (zh) |
IL (1) | IL196652A0 (zh) |
IT (1) | ITMI20061474A1 (zh) |
MX (1) | MX2009000828A (zh) |
NO (1) | NO342506B1 (zh) |
PL (1) | PL2044080T3 (zh) |
PT (1) | PT2044080T (zh) |
RU (1) | RU2450007C2 (zh) |
SI (1) | SI2044080T1 (zh) |
WO (1) | WO2008012003A1 (zh) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9379559B2 (en) | 2012-02-03 | 2016-06-28 | International Business Machines Corporation | System and method of charging a vehicle using a dynamic power grid, and system and method of managing power consumption in the vehicle |
PL235836B1 (pl) | 2012-10-25 | 2020-11-02 | Inst Chemii Organicznej Polskiej Akademii Nauk | Pochodne kamptotecyny, sposób ich otrzymywania i zastosowanie |
CN108690036B (zh) * | 2018-05-22 | 2021-05-25 | 北京海步医药科技股份有限公司 | 一种10-二氟甲基喜树碱类化合物及其制备方法和应用 |
LU102067B1 (en) | 2020-09-17 | 2022-03-18 | Narodowy Inst Lekow | 7-Ethyl-10-hydroxycamptothecin derivatives for use in the treatment of cancer |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5972955A (en) * | 1995-06-06 | 1999-10-26 | Dr. Reddy's Research Foundation | Water soluble C-ring analogues of 20(S)-camptothecin |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
IT1282673B1 (it) * | 1996-02-23 | 1998-03-31 | Ist Naz Stud Cura Dei Tumori | Derivati della camptotecina e loro uso come agenti antitumorali |
-
2006
- 2006-07-26 IT IT001474A patent/ITMI20061474A1/it unknown
-
2007
- 2007-07-16 US US12/373,834 patent/US8193210B2/en active Active
- 2007-07-16 AU AU2007278509A patent/AU2007278509B2/en not_active Ceased
- 2007-07-16 KR KR1020097001596A patent/KR101496374B1/ko active IP Right Grant
- 2007-07-16 PT PT77860963T patent/PT2044080T/pt unknown
- 2007-07-16 DK DK07786096.3T patent/DK2044080T3/en active
- 2007-07-16 ES ES07786096.3T patent/ES2628022T3/es active Active
- 2007-07-16 EP EP07786096.3A patent/EP2044080B1/en active Active
- 2007-07-16 WO PCT/EP2007/006294 patent/WO2008012003A1/en active Application Filing
- 2007-07-16 PL PL07786096T patent/PL2044080T3/pl unknown
- 2007-07-16 MX MX2009000828A patent/MX2009000828A/es active IP Right Grant
- 2007-07-16 CA CA2658902A patent/CA2658902C/en not_active Expired - Fee Related
- 2007-07-16 CN CN2007800281978A patent/CN101495485B/zh active Active
- 2007-07-16 JP JP2009521140A patent/JP5313895B2/ja not_active Expired - Fee Related
- 2007-07-16 RU RU2009102241/04A patent/RU2450007C2/ru active
- 2007-07-16 SI SI200731933A patent/SI2044080T1/sl unknown
- 2007-07-16 BR BRPI0714672A patent/BRPI0714672B8/pt not_active IP Right Cessation
-
2009
- 2009-01-16 NO NO20090252A patent/NO342506B1/no not_active IP Right Cessation
- 2009-01-22 IL IL196652A patent/IL196652A0/en unknown
-
2010
- 2010-01-15 HK HK10100447.7A patent/HK1135692A1/xx not_active IP Right Cessation
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5972955A (en) * | 1995-06-06 | 1999-10-26 | Dr. Reddy's Research Foundation | Water soluble C-ring analogues of 20(S)-camptothecin |
Non-Patent Citations (4)
Title |
---|
Duvvuri Subrahmanyam, et al..Novel C-Ring Analogues of 20(S)-camptothecin. Part 3: Synthesis and Their In Vitro Cytotoxicity of A-, B- and C-ring Analogues.《Bioorganic & Medicinal Chemistry Letters》.Elsevier Science Ltd.,2000,第10卷(第4期),369-371. |
Duvvuri Subrahmanyam, et al..Novel C-Ring Analogues of 20(S)-camptothecin. Part 3: Synthesis and Their In Vitro Cytotoxicity of A-, B- and C-ring Analogues.《Bioorganic & * |
Medicinal Chemistry Letters》.Elsevier Science Ltd.,2000,第10卷(第4期),369-371. * |
Thierry Brunin, et al..Towards new camptothecins. Part 3: Synthesis of 5-methoxycarbonyl camptothecin.《Tetrahedron》.Elsevier Ltd.,2006,第62卷(第17期),3959–3968. * |
Also Published As
Publication number | Publication date |
---|---|
CN101495485A (zh) | 2009-07-29 |
NO342506B1 (no) | 2018-06-04 |
CA2658902A1 (en) | 2008-01-31 |
KR20090032100A (ko) | 2009-03-31 |
NO20090252L (no) | 2009-01-16 |
RU2450007C2 (ru) | 2012-05-10 |
CA2658902C (en) | 2014-09-23 |
PL2044080T3 (pl) | 2017-10-31 |
PT2044080T (pt) | 2017-06-15 |
JP5313895B2 (ja) | 2013-10-09 |
KR101496374B1 (ko) | 2015-03-04 |
EP2044080B1 (en) | 2017-05-10 |
US8193210B2 (en) | 2012-06-05 |
US20100010032A1 (en) | 2010-01-14 |
ES2628022T3 (es) | 2017-08-01 |
BRPI0714672B8 (pt) | 2021-05-25 |
WO2008012003A1 (en) | 2008-01-31 |
RU2009102241A (ru) | 2010-07-27 |
HK1135692A1 (en) | 2010-06-11 |
AU2007278509B2 (en) | 2012-10-04 |
DK2044080T3 (en) | 2017-07-17 |
EP2044080A1 (en) | 2009-04-08 |
BRPI0714672A2 (pt) | 2013-03-26 |
BRPI0714672B1 (pt) | 2020-10-20 |
IL196652A0 (en) | 2009-11-18 |
SI2044080T1 (sl) | 2017-07-31 |
JP2010500971A (ja) | 2010-01-14 |
MX2009000828A (es) | 2009-02-03 |
AU2007278509A1 (en) | 2008-01-31 |
ITMI20061474A1 (it) | 2008-01-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN101495486B (zh) | 具有抗肿瘤活性的喜树碱衍生物 | |
CN101495485B (zh) | 具有抗肿瘤活性的喜树碱衍生物 | |
JP5198447B2 (ja) | 抗腫瘍活性を有するカンプトテシン誘導体 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: DE Ref document number: 1135692 Country of ref document: HK |
|
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
REG | Reference to a national code |
Ref country code: HK Ref legal event code: GR Ref document number: 1135692 Country of ref document: HK |