CN111918672B - 人工合成的鞘氨醇衍生物类脂质单体及其递送核酸的用途 - Google Patents
人工合成的鞘氨醇衍生物类脂质单体及其递送核酸的用途 Download PDFInfo
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- CN111918672B CN111918672B CN201980023106.4A CN201980023106A CN111918672B CN 111918672 B CN111918672 B CN 111918672B CN 201980023106 A CN201980023106 A CN 201980023106A CN 111918672 B CN111918672 B CN 111918672B
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Abstract
本发明提供了人工合成的鞘氨醇类脂质单体及其递送核酸的用途。具体地,本发明提供了使用式I化合物、其立体异构体或其药学上可接受的盐或包含式I的化合物、其立体异构体或其药学上可接受的盐的组合对细胞或受试者递送核酸的用途或方法,(I)
Description
本申请要求2018年3月29日提交的申请号PCT/CN2018/081155的发明名称为“化合物或中药提取物在制备核酸递送试剂中的应用及其相关产品”的PCT申请的优先权,其全文通过引用并入本文。
技术领域
本发明一般涉及核酸治疗递送方式,更具体涉及小RNA更加高效的递送载体和递送方式。
背景技术
核酸作为生命体主要的遗传物质,也具有其独特的药物研发潜能,目前,FDA获批的核酸类药物共有6种,包括:福米韦生(ISIS PHARMS INC,NDA:20-961)主要用于巨细胞病毒性视网膜炎的治疗;哌加他尼(VALEANT PHARMS LLC,NDA:21-756)主要用药于新血管年龄相关性黄斑变性;米泊美生(KASTLE THERAPS LLC,NDA:203458)常用于纯合子家族性高胆固醇血症;Exondys 51(SAREPTA THERAPS INC,NDA:206488)用于杜氏肌营养不良;去纤维钠(JAZZ PHAEMS INC,NDA:208114)治疗造血后伴有肾或肺功能障碍的肝静脉闭塞性疾病;Nusinersen(BIOGEN IDEC,NDA:209531)治疗脊髓性肌肉萎缩症;Patisiran(ALNYLAMPHARMS INC,NDA:210922)用于与遗传性甲亢蛋白有关的淀粉样变性疾病。而这些药物的给药方式均为注射用药,且药物递送效率也较为低下。
在我们之前的研究中可以看出,植物汤剂体中含有数百种脂质单体。本申请在进一步实验中发现,鞘氨醇类脂质在递送过程中具有更优的效率,故人工合成了一系列鞘氨醇类脂质及相关衍生物,针对性地对鞘氨醇类脂质的递送效果进行探究,得出鞘氨醇及其衍生物类脂质可以有效递送小RNA的结论。
发明内容
本申请部分基于发明人对一系列鞘氨醇类脂质单体的发现。目的在于发现了鞘氨醇单体能够作为载体,高效地将小RNA递送进入细胞内。
本项发明的亮点是发现了鞘氨醇类脂质单体,在对机体无毒的前提下能够高效的将小RNA作为药物递送入细胞内,极大地提高了核酸类药物的递送效率和药用价值。
鞘氨醇类脂质及其衍生物单体能够有效递送sRNA进入细胞,用带FAM荧光标记的功能性小RNA(sRNA-PGY6)作为标志物检测递送至细胞后荧光偏移,与自由摄取相比,鞘氨醇类脂质及其衍生物单体递送小RNA实验组有明显的荧光偏移,表示鞘氨醇类脂质及其衍生物单体形成本草体能更有效的将小RNA递送至细胞内从而发挥相应功能。
本发明提供了以下内容:
1.使用式I化合物、其立体异构体或其药学上可接受的盐或包含式I的化合物、其立体异构体或其药学上可接受的盐的组合对细胞或受试者递送核酸的用途或方法,
其中:
R1选自C1-20烷基或C2-20烯基,其任选被一至三个羟基取代;
R2为氢,且R3为羟基;或者
R2与R3一起形成氧代(=O);
R4与R5独立选自氢、C1-20烷基、C1-30烷基酰基和C1-30烯基酰基,所述C1-30烷基酰基和C1-30烯基酰基任选被一至三个以下基团取代:生物素酰基、羟基;
或-NR4R5基团一起为季铵阳离子;
R6选自氢、羟基、磷酸酯基、-O-糖基、神经节苷酯基(Ganglioside)、氨基乙氧基膦酸酯基(NH2-CH2-CH2-O-P(O)OH-)。
2.项1的用途或方法,其中:
R1选自C14-20烷基或含有一个双键的C14-20烯基;
R4与R5独立选自氢、C1-6烷基、C6-30烷基酰基和C6-30烯基酰基,所述C6-30烷基酰基和C6-30烯基酰基任选在末端被一个以下基团取代:生物素酰基、羟基,或任选在酰基的α-碳被一个羟基取代。
3.项1或2的用途或方法,其中R4与R5各自独立选自甲基。
4.项1或2的用途或方法,其中R4与R5中至少一个为氢且另一个为直链C6-30烷基酰基或直链C6-30烯基酰基。
5.项1-4中任一项的用途或方法,其中R4与R5中至少一个为氢且另一个为直链C7-14烷基;或者R4与R5都为C1-6烷基。
6.项1-5中任一项的用途或方法,其中R1选自直链C14-20烷基或含有一个双键的直链C14-20烯基。
7.项1-6中任一项的用途或方法,其中所述糖基为葡糖基、乳糖基(Lactosyl)、半乳糖基(galactosyl)。
8.项1-7中任一项的用途或方法,其中-NR4R5基团一起为-NH3 +、N(CH3)3 +。
9.项1-8中任一项的用途或方法,其中所述糖基为1-β-D-葡糖基;
R2为氢,且R3为羟基;
R1选自含有一个双键的C14-20烯基,且所述烯基紧邻R2和R3所连接的碳原子。
10.前述项中任一项的用途或方法,其中所述化合物具有下式Ia:
11.使用选自下组的化合物、其立体异构体或其药学上可接受的盐或包含所述化合物、其立体异构体或其药学上可接受的盐的组合对细胞或受试者递送核酸的用途或方法:
12.前述项中任一项的用途或方法,其中所述组合是包含No.41、No.38、No.48中任一种或多种的组合,包含No.41、No.38、No.48中任一种或多种和选自项11中的任一种或多种化合物的组合,包含以下各项的组合:No.41+No.38+鞘氨醇衍生物So-1;No.41+No.38+鞘氨醇衍生物So-2;No.41+No.38+鞘氨醇衍生物So-3;No.41+No.38+鞘氨醇衍生物So-4;No.41+No.38+鞘氨醇衍生物So-5;No.41+No.38+鞘氨醇衍生物So-6;No.41+No.38+鞘氨醇衍生物So-7;No.41+No.38+鞘氨醇衍生物So-8;No.41+No.38+鞘氨醇衍生物So-9;No.41+No.38+鞘氨醇衍生物So-10;No.41+No.38+鞘氨醇衍生物So-11;No.41+No.38+鞘氨醇衍生物So-12;No.41+No.38+鞘氨醇衍生物So-13;No.41+No.38+鞘氨醇衍生物So-14;No.41+No.38+鞘氨醇衍生物So-15;No.41+No.38+鞘氨醇衍生物So-45;No.41+No.38+鞘氨醇衍生物So-46;No.41+No.38+鞘氨醇衍生物So-47;No.41+No.38+鞘氨醇衍生物So-48;No.41+No.38+鞘氨醇衍生物So-49;No.41+No.38+鞘氨醇衍生物So-50;No.41+No.38+鞘氨醇衍生物So-51;No.41+No.38+鞘氨醇衍生物So-52;No.41+No.38+鞘氨醇衍生物So-53;No.41+No.38+鞘氨醇衍生物So-54;No.41+No.38+鞘氨醇衍生物So-55;No.41+No.38+鞘氨醇衍生物So-56;No.41+No.38+鞘氨醇衍生物So-57;No.41+No.38+鞘氨醇衍生物So-58;No.41+No.38+鞘氨醇衍生物So-59;No.41+No.38+鞘氨醇衍生物So-60;No.41+No.38+鞘氨醇衍生物So-61;No.41+No.38+鞘氨醇衍生物So-62;No.41+No.38+鞘氨醇衍生物So-63;No.41+No.38+鞘氨醇衍生物So-64;No.41+No.38+鞘氨醇衍生物So-65;No.41+No.38+鞘氨醇衍生物So-66;No.41+No.38+鞘氨醇衍生物So-67;No.41+No.38+鞘氨醇衍生物So-68;No.41+No.38+鞘氨醇衍生物So-69;No.41+No.38+鞘氨醇衍生物So-70;No.41+No.38+鞘氨醇衍生物So-71;No.41+No.38+鞘氨醇衍生物So-72;No.41+No.38+鞘氨醇衍生物So-73;No.41+No.38+鞘氨醇衍生物So-74;No.41+No.38+鞘氨醇衍生物So-75;No.41+No.38+No.48+鞘氨醇衍生物So-42;No.41+No.38+No.48+鞘氨醇衍生物So-43;No.41+No.38+No.48+鞘氨醇衍生物So-44;No.41+No.38+No.48+鞘氨醇衍生物So-45;No.41+No.38+No.48+鞘氨醇衍生物So-46;No.41+No.38+No.48+鞘氨醇衍生物So-47;No.41+No.38+No.48+鞘氨醇衍生物So-52;No.41+No.38+No.48+鞘氨醇衍生物So-56;No.41+No.38+No.48+鞘氨醇衍生物So-57;No.41+No.38+No.48+鞘氨醇衍生物So-58;No.41+No.38+No.48+鞘氨醇衍生物So-59;No.41+No.38+No.48+鞘氨醇衍生物So-60;No.41+No.38+No.48+鞘氨醇衍生物So-61;No.41+No.38+No.48+鞘氨醇衍生物So-62;No.41+No.38+No.48+鞘氨醇衍生物So-63;No.41+No.38+No.48+鞘氨醇衍生物So-64;No.41+No.38+No.48+鞘氨醇衍生物So-67;No.41+No.38+No.48+鞘氨醇衍生物So-68;No.41+No.38+No.48+鞘氨醇衍生物So-69;No.41+No.38+No.48+鞘氨醇衍生物So-70;No.41+No.38+鞘氨醇衍生物So-23、25、26、27、29、30、31、32、33、34、35、36、37、38、40、41、42、43、44、70、71或73中任一种或多种;优选地,其中上述化合物的使用浓度如表1所述;优选地,上述各种化合物的比例为0.1-10∶0.1-10,0.2-9∶0.2-9,0.3-8∶0.3-8;0.4-7∶0.4-7;0.5-6∶0.5-6;0.6-5∶0.6-5;0.7-4∶0.7-4;0.8-3∶0.8-3;0.9-2∶0.9-2;更优选1∶1。
13.前述项中任一项的用途或方法,其中所述核酸是合成或纯化的、治疗性或非治疗性、和/或诊断性或非诊断性的,例如选自RNA和DNA,例如选自单链或双链或部分双链的RNA和DNA;
优选地,其中在核酸是治疗性或诊断性时,所述核酸用于治疗或诊断选自下组的疾病:炎性疾病、肺炎、心肌炎、急慢性胃炎、急慢性肠炎、急慢性肝炎、急慢性肾炎、皮炎、脑炎、淋巴炎、结膜炎、角膜炎、虹膜睫状体炎、中耳炎、过敏性鼻炎、哮喘、肺纤维化慢性阻塞性肺疾病、过敏性皮炎、镰状细胞病、多发性硬化、***性红斑狼疮、狼疮性肾炎、肺癌、胃癌、结直肠癌、肝癌、胰腺癌、***、乳腺癌、白血病、多发性骨髓瘤、糖尿病和痛风。
14.项13的用途或方法,其中所述RNA选自:信使RNA(mRNA)、rRNA(核糖体RNA)、tRNA(转运RNA)、不均一核RNA(hnRNA)、小核RNA(snRNA)、核仁小RNA(snoRNA)、小胞质RNA、小RNA、转移-信使RNA(tmRNA)、端粒酶RNA和反义RNA,优选小RNA。
15.项14的用途或方法,其中所述小RNA长度是14-32bp、16-28bp或18-24bp。
16.项15的用途或方法,其中所述小RNA包含GTTCAGAGTTCTACAGTCCGA的序列。
17.前述项中任一项的用途或方法,其中所述递送包括通过加热法、逆向蒸发法、直接混合、反复冻融和/或薄膜分散处理所述化合物、其立体异构体或其药学上可接受的盐或包含式I的化合物、其立体异构体或其药学上可接受的盐或包含它们的组合。
18.项17的用途或方法,其中所述加热法的温度为约0℃至约100℃,约25℃至约100℃,优选约80℃至约100℃,例如4℃,37℃,60℃,80℃或100℃;所述逆向蒸发法的制备温度为约25℃至约70℃,优选约55℃,加热时间为约0分钟-约24小时,约5分钟-约20小时,约5分钟-约16小时,约5分钟-约10小时,约5分钟-约4小时,或者约10小时-约1小时,优选15分钟。
19.项17的用途,其中所述递送包括体外细胞递送,或受试者体内递送。
20项19的用途或方法,其中所述受试者体内递送包括口服、静脉内施用,如推注或灌注,通过皮下、肌肉内、动脉内、腹膜内、肺内、脑脊髓内、关节内、滑膜内、鞘内、损伤内、和/或吸入路径如鼻内,通常通过静脉内或皮下施用。
21.式I化合物、其立体异构体或其药学上可接受的盐或包含式I的化合物、其立体异构体或其药学上可接受的盐的组合在制备核酸递送的试剂中的用途,
其中:
R1选自C1-20烷基或C2-20烯基,其任选被一至三个羟基取代;
R2为氢,且R3为羟基;或者
R2与R3一起形成氧代(=O);
R4与R5独立选自氢、C1-20烷基、C1-30烷基酰基和C1-30烯基酰基,所述C1-30烷基酰基和C1-30烯基酰基任选被一至三个以下基团取代:生物素酰基、羟基;
或-NR4R5基团一起为季铵阳离子;
R6选自氢、羟基、磷酸酯基、-O-糖基、神经节苷酯基(Ganglioside)、氨基乙氧基膦酸酯基(NH2-CH2-CH2-O-P(O)OH-)。
22.项21的用途,其中
R1选自C14-20烷基或含有一个双键的C14-20烯基;
R4与R5独立选自氢、C1-6烷基、C6-30烷基酰基和C6-30烯基酰基,所述C6-30烷基酰基和C6-30烯基酰基任选在末端被一个以下基团取代:生物素酰基、羟基,或任选在酰基的α-碳被一个羟基取代。
23.项21或22的用途,其中R4与R5各自独立选自甲基。
24.项21或22的用途,其中R4与R5中至少一个为氢且另一个为直链C6-30烷基酰基或直链C6-30烯基酰基。
25.项21-24中任一项的用途,其中R4与R5中至少一个为氢且另一个为直链C7-14烷基;或者R4与R5都为C1-6烷基。
26.项21-25中任一项的用途,其中R1选自直链C14-20烷基或含有一个双键的直链C14-20烯基。
27.项21-26中任一项的用途,其中所述糖基为葡糖基、乳糖基(Lactosyl)、半乳糖基(galactosyl)。
28.项21-27中任一项的用途,其中所述糖基为1-β-D-葡糖基;
R2为氢,且R3为羟基;
R1选自含有一个双键的C14-20烯基,且所述烯基紧邻R2和R3所连接的碳原子。
29.项21-28中任一项的用途,其中所述化合物具有下式Ia:
30.项21-29中任一项的用途,所述式I的化合物选自项11中的一种或多种化合物。
31.项21-30中任一项的用途,其中所述组合是包含No.41、No.38、No.48中任一种或多种的组合,包含No.41、No.38、No.48中任一种或多种和选自项7中的任一种或多种化合物的组合,包含以下各项的组合:No.41+No.38+鞘氨醇衍生物So-1;No.41+No.38+鞘氨醇衍生物So-2;No.41+No.38+鞘氨醇衍生物So-3;No.41+No.38+鞘氨醇衍生物So-4;No.41+No.38+鞘氨醇衍生物So-5;No.41+No.38+鞘氨醇衍生物So-6;No.41+No.38+鞘氨醇衍生物So-7;No.41+No.38+鞘氨醇衍生物So-8;No.41+No.38+鞘氨醇衍生物So-9;No.41+No.38+鞘氨醇衍生物So-10;No.41+No.38+鞘氨醇衍生物So-11;No.41+No.38+鞘氨醇衍生物So-12;No.41+No.38+鞘氨醇衍生物So-13;No.41+No.38+鞘氨醇衍生物So-14;No.41+No.38+鞘氨醇衍生物So-15;No.41+No.38+鞘氨醇衍生物So-45;No.41+No.38+鞘氨醇衍生物So-46;No.41+No.38+鞘氨醇衍生物So-47;No.41+No.38+鞘氨醇衍生物So-48;No.41+No.38+鞘氨醇衍生物So-49;No.41+No.38+鞘氨醇衍生物So-50;No.41+No.38+鞘氨醇衍生物So-51;No.41+No.38+鞘氨醇衍生物So-52;No.41+No.38+鞘氨醇衍生物So-53;No.41+No.38+鞘氨醇衍生物So-54;No.41+No.38+鞘氨醇衍生物So-55;No.41+No.38+鞘氨醇衍生物So-56;No.41+No.38+鞘氨醇衍生物So-57;No.41+No.38+鞘氨醇衍生物So-58;No.41+No.38+鞘氨醇衍生物So-59;No.41+No.38+鞘氨醇衍生物So-60;No.41+No.38+鞘氨醇衍生物So-61;No.41+No.38+鞘氨醇衍生物So-62;No.41+No.38+鞘氨醇衍生物So-63;No.41+No.38+鞘氨醇衍生物So-64;No.41+No.38+鞘氨醇衍生物So-65;No.41+No.38+鞘氨醇衍生物So-66;No.41+No.38+鞘氨醇衍生物So-67;No.41+No.38+鞘氨醇衍生物So-68;No.41+No.38+鞘氨醇衍生物So-69;No.41+No.38+鞘氨醇衍生物So-70;No.41+No.38+鞘氨醇衍生物So-71;No.41+No.38+鞘氨醇衍生物So-72;No.41+No.38+鞘氨醇衍生物So-73;No.41+No.38+鞘氨醇衍生物So-74;No.41+No.38+鞘氨醇衍生物So-75;No.41+No.38+No.48+鞘氨醇衍生物So-42;No.41+No.38+No.48+鞘氨醇衍生物So-43;No.41+No.38+No.48+鞘氨醇衍生物So-44;No.41+No.38+No.48+鞘氨醇衍生物So-45;No.41+No.38+No.48+鞘氨醇衍生物So-46;No.41+No.38+No.48+鞘氨醇衍生物So-47;No.41+No.38+No.48+鞘氨醇衍生物So-52;No.41+No.38+No.48+鞘氨醇衍生物So-56;No.41+No.38+No.48+鞘氨醇衍生物So-57;No.41+No.38+No.48+鞘氨醇衍生物So-58;No.41+No.38+No.48+鞘氨醇衍生物So-59;No.41+No.38+No.48+鞘氨醇衍生物So-60;No.41+No.38+No.48+鞘氨醇衍生物So-61;No.41+No.38+No.48+鞘氨醇衍生物So-62;No.41+No.38+No.48+鞘氨醇衍生物So-63;No.41+No.38+No.48+鞘氨醇衍生物So-64;No.41+No.38+No.48+鞘氨醇衍生物So-67;No.41+No.38+No.48+鞘氨醇衍生物So-68;No.41+No.38+No.48+鞘氨醇衍生物So-69;No.41+No.38+No.48+鞘氨醇衍生物So-70;No.41+No.38+鞘氨醇衍生物So-23、25、26、27、29、30、31、32、33、34、35、36、37、38、40、41、42、43、44、70、71或73中任一种或多种;优选地,其中上述化合物的使用浓度如表1所述;优选地,上述各种化合物的比例为0.1-10∶0.1-10,0.2-9∶0.2-9,0.3-8∶0.3-8;0.4-7∶0.4-7;0.5-6∶0.5-6;0.6-5∶0.6-5;0.7-4∶0.7-4;0.8-3∶0.8-3;0.9-2∶0.9-2;更优选1∶1。
32.项21-31中任一项的用途,其中所述核酸是合成或纯化的、治疗性或非治疗性、和/或诊断性或非诊断性的,例如选自RNA和DNA,例如选自单链或双链或部分双链的RNA和DNA;
优选地,其中在核酸是治疗性或诊断性时,所述核酸用于治疗或诊断选自下组的疾病:炎性疾病、肺炎、心肌炎、急慢性胃炎、急慢性肠炎、急慢性肝炎、急慢性肾炎、皮炎、脑炎、淋巴炎、结膜炎、角膜炎、虹膜睫状体炎、中耳炎、过敏性鼻炎、哮喘、肺纤维化慢性阻塞性肺疾病、过敏性皮炎、镰状细胞病、多发性硬化、***性红斑狼疮、狼疮性肾炎、肺癌、胃癌、结直肠癌、肝癌、胰腺癌、***、乳腺癌、白血病、多发性骨髓瘤、糖尿病和痛风。
33.项32的用途,其中所述RNA选自:信使RNA(mRNA)、rRNA(核糖体RNA)、tRNA(转运RNA)、不均一核RNA(hnRNA)、小核RNA(snRNA)、核仁小RNA(snoRNA)、小胞质RNA、小RNA、转移-信使RNA(tmRNA)、端粒酶RNA和反义RNA,优选小RNA。
34.项33的用途,其中所述小RNA长度是14-32bp、16-28bp或18-24bp。
35.项34的用途,其中所述小RNA包含选自下组的序列:GTTCAGAGTTCTACAGTCCGA。
36.项21-35中任一项的用途,其中所述递送包括通过加热法、逆向蒸发法和/或直接混合处理所述化合物、其立体异构体或其药学上可接受的盐或包含式I的化合物、其立体异构体或其药学上可接受的盐或包含它们的组合和核酸的步骤。
37.项36的用途,其中所述加热法的温度为约0℃至约100℃,约25℃至约100℃,优选约80℃至约100℃,例如4℃,37℃,60℃,80℃或100℃;所述逆向蒸发法的制备温度为约25℃至约70℃,优选约55℃,加热时间为约0分钟-约24小时,约5分钟-约20小时,约5分钟-约16小时,约5分钟-约10小时,约5分钟-约4小时,或者约10小时-约1小时,优选15分钟。
38.项36的用途,其中所述递送包括体外细胞递送,或受试者体内递送。
39.项38的用途,其中所述受试者体内递送包括口服、静脉内施用,如推注或灌注,通过皮下、肌肉内、动脉内、腹膜内、肺内、脑脊髓内、关节内、滑膜内、鞘内、损伤内、和/或吸入路径如鼻内,通常通过静脉内或皮下施用。
40.组合物或化合物组合,其包含项1-12中所述的化合物、其立体异构体或其药学上可接受的盐或包含它们的组合。
41.项40的组合物或化合物组合,其用于对细胞或受试者递送核酸。
42.项41的组合物或化合物组合,其中所述核酸是合成或纯化的、治疗性或非治疗性、和/或诊断性或非诊断性的,例如选自RNA和DNA,例如选自单链或双链或部分双链的RNA和DNA;
优选地,其中在核酸是治疗性或诊断性时,所述核酸用于治疗或诊断选自下组的疾病:炎性疾病、肺炎、心肌炎、急慢性胃炎、急慢性肠炎、急慢性肝炎、急慢性肾炎、皮炎、脑炎、淋巴炎、结膜炎、角膜炎、虹膜睫状体炎、中耳炎、过敏性鼻炎、哮喘、肺纤维化慢性阻塞性肺疾病、过敏性皮炎、镰状细胞病、多发性硬化、***性红斑狼疮、狼疮性肾炎、肺癌、胃癌、结直肠癌、肝癌、胰腺癌、***、乳腺癌、白血病、多发性骨髓瘤、糖尿病和痛风。
43.项42的组合物或化合物组合,其中所述RNA选自:信使RNA(mRNA)、rRNA(核糖体RNA)、tRNA(转运RNA)、不均一核RNA(hnRNA)、小核RNA(snRNA)、核仁小RNA(snoRNA)、小胞质RNA、小RNA、转移-信使RNA(tmRNA)、端粒酶RNA和反义RNA,优选小RNA。
44.项43的组合物或化合物组合,其中所述小RNA长度是14-32bp、16-28bp或18-24bp。
45.项44的组合物或化合物组合,其中所述小RNA包含GTTCAGAGTTCTACAGTCCGA的序列。
46.项40-45中任一项的组合物或化合物组合,其中所述递送包括通过加热法、逆向蒸发法和/或直接混合处理所述化合物、其立体异构体或其药学上可接受的盐或包含式I的化合物、其立体异构体或其药学上可接受的盐的组合和核酸的步骤。
47.项46的组合物或化合物组合,其中所述加热法的温度为约0℃至约100℃,约25℃至约100℃,优选约80℃至约100℃,例如4℃,37℃,60℃,80℃或100℃;所述逆向蒸发法的制备温度为约25℃至约70℃,优选约55℃。加热时间为约0分钟-约24小时,约5分钟-约20小时,约5分钟-约16小时,约5分钟-约10小时,约5分钟-约4小时,或者约10小时-约1小时,优选15分钟。
48.项46的组合物或化合物组合,其中所述递送包括体外细胞递送,或受试者体内递送。
49.项48的组合物或化合物组合,其中所述受试者体内递送包括口服、静脉内施用,如推注或灌注,通过皮下、肌肉内、动脉内、腹膜内、肺内、脑脊髓内、关节内、滑膜内、鞘内、损伤内、和/或吸入路径如鼻内,通常通过静脉内或皮下施用。
50.试剂盒,其包含项1-12中所述的化合物其立体异构体或其药学上可接受的盐或包含它们的组合;优选地,所述试剂盒用于上述任一项的用途。
附图说明
图1a-e:不同浓度鞘氨醇衍生物So-1递送单链PGY-sRNA-6进入293T细胞;
图2a-e:不同浓度鞘氨醇衍生物So-3递送单链PGY-sRNA-6进入293T细胞;
图3a-e:不同浓度鞘氨醇衍生物So-4递送单链PGY-sRNA-6进入293T细胞;
图4a-e:不同浓度鞘氨醇衍生物So-5递送单链PGY-sRNA-6进入293T细胞;
图5a-e:不同浓度鞘氨醇衍生物So-7递送单链PGY-sRNA-6进入293T细胞;
图6a-e:不同浓度鞘氨醇衍生物So-8递送单链PGY-sRNA-6进入293T细胞;
图7a-e:不同浓度鞘氨醇衍生物So-9递送单链PGY-sRNA-6进入293T细胞;
图8a-e:不同浓度鞘氨醇衍生物So-10递送单链PGY-sRNA-6进入293T细胞;
图9a-e:不同浓度鞘氨醇衍生物So-11递送单链PGY-sRNA-6进入293T细胞;
图10a-e:不同浓度鞘氨醇衍生物So-12递送单链PGY-sRNA-6进入293T细胞;
图11a-e:不同浓度鞘氨醇衍生物So-13递送单链PGY-sRNA-6进入293T细胞;
图12a-e:不同浓度鞘氨醇衍生物So-14递送单链PGY-sRNA-6进入293T细胞;
图13a-e:不同浓度鞘氨醇衍生物So-15递送单链PGY-sRNA-6进入293T细胞;
图14a-e:不同浓度鞘氨醇衍生物So-26递送单链PGY-sRNA-6进入293T细胞;
图15a-e:不同浓度鞘氨醇衍生物So-46递送单链PGY-sRNA-6进入293T细胞;
图16a-e:不同浓度鞘氨醇衍生物So-49递送单链PGY-sRNA-6进入293T细胞;
图17a-e:不同浓度鞘氨醇衍生物So-53递送单链PGY-sRNA-6进入293T细胞;
图18a-e:不同浓度鞘氨醇衍生物So-60递送单链PGY-sRNA-6进入293T细胞;
图19a-e:不同浓度鞘氨醇衍生物So-61递送单链PGY-sRNA-6进入293T细胞;
图20a-e:不同浓度鞘氨醇衍生物So-62递送单链PGY-sRNA-6进入293T细胞;
图21a-e:不同浓度鞘氨醇衍生物So-63递送单链PGY-sRNA-6进入293T细胞;
图22a-e:不同浓度鞘氨醇衍生物So-64递送单链PGY-sRNA-6进入293T细胞;
图23a-e:不同浓度鞘氨醇衍生物So-65递送单链PGY-sRNA-6进入293T细胞;
图24a-e:不同浓度鞘氨醇衍生物So-66递送单链PGY-sRNA-6进入293T细胞;
图25a-e:不同浓度鞘氨醇衍生物So-67递送单链PGY-sRNA-6进入293T细胞;
图26a-e:不同浓度鞘氨醇衍生物So-68递送单链PGY-sRNA-6进入293T细胞;
图27a-e:不同浓度鞘氨醇衍生物So-69递送单链PGY-sRNA-6进入293T细胞;
图28a-e:不同浓度鞘氨醇衍生物So-70递送单链PGY-sRNA-6进入293T细胞;
图29a-e:不同浓度鞘氨醇衍生物So-71递送单链PGY-sRNA-6进入293T细胞;
图30a-e:不同浓度鞘氨醇衍生物So-72递送单链PGY-sRNA-6进入293T细胞;
图31a-e:不同浓度鞘氨醇衍生物So-73递送单链PGY-sRNA-6进入293T细胞;
图32a-e:不同浓度鞘氨醇衍生物So-74递送单链PGY-sRNA-6进入293T细胞;
图33a-e:不同浓度鞘氨醇衍生物So-75递送单链PGY-sRNA-6进入293T细胞;
图34a-c:No.41+No.38+鞘氨醇衍生物So-1混合物递送单链PGY-sRNA-6进入THP-1细胞;
图35a-c:No.41+No.38+鞘氨醇衍生物So-2混合物递送单链PGY-sRNA-6进入THP-1细胞;
图36a-c:No.41+No.38+鞘氨醇衍生物So-3混合物递送单链PGY-sRNA-6进入THP-1细胞;
图37a-c:No.41+No.38+鞘氨醇衍生物So-4混合物递送单链PGY-sRNA-6进入THP-1细胞;
图38a-c:No.41+No.38+鞘氨醇衍生物So-5混合物递送单链PGY-sRNA-6进入THP-1细胞;
图39a-c:No.41+No.38+鞘氨醇衍生物So-6混合物递送单链PGY-sRNA-6进入THP-1细胞;
图40a-c:No.41+No.38+鞘氨醇衍生物So-7混合物递送单链PGY-sRNA-6进入THP-1细胞;
图41a-c:No.41+No.38+鞘氨醇衍生物So-8混合物递送单链PGY-sRNA-6进入THP-1细胞;
图42a-c:No.41+No.38+鞘氨醇衍生物So-9混合物递送单链PGY-sRNA-6进入THP-1细胞;
图43a-c:No.41+No.38+鞘氨醇衍生物So-10混合物递送单链PGY-sRNA-6进入THP-1细胞;
图44a-c:No.41+No.38+鞘氨醇衍生物So-11混合物递送单链PGY-sRNA-6进入THP-1细胞;
图45a-c:No.41+No.38+鞘氨醇衍生物So-12混合物递送单链PGY-sRNA-6进入THP-1细胞;
图46a-c:No.41+No.38+鞘氨醇衍生物So-13混合物递送单链PGY-sRNA-6进入THP-1细胞;
图47a-c:No.41+No.38+鞘氨醇衍生物So-14混合物递送单链PGY-sRNA-6进入THP-1细胞;
图48a-c:No.41+No.38+鞘氨醇衍生物So-15混合物递送单链PGY-sRNA-6进入THP-1细胞;
图49a-c:No.41+No.38+鞘氨醇衍生物So-45混合物递送单链PGY-sRNA-6进入THP-1细胞;
图50a-c:No.41+No.38+鞘氨醇衍生物So-46混合物递送单链PGY-sRNA-6进入THP-1细胞;
图51a-c:No.41+No.38+鞘氨醇衍生物So-47混合物递送单链PGY-sRNA-6进入THP-1细胞;
图52a-c:No.41+No.38+鞘氨醇衍生物So-48混合物递送单链PGY-sRNA-6进入THP-1细胞;
图53a-c:No.41+No.38+鞘氨醇衍生物So-49混合物递送单链PGY-sRNA-6进入THP-1细胞;
图54a-c:No.41+No.38+鞘氨醇衍生物So-50混合物递送单链PGY-sRNA-6进入THP-1细胞;
图55a-c:No.41+No.38+鞘氨醇衍生物So-51混合物递送单链PGY-sRNA-6进入THP-1细胞;
图56a-c:No.41+No.38+鞘氨醇衍生物So-52混合物递送单链PGY-sRNA-6进入THP-1细胞;
图57a-c:No.41+No.38+鞘氨醇衍生物So-53混合物递送单链PGY-sRNA-6进入THP-1细胞;
图58a-c:No.41+No.38+鞘氨醇衍生物So-54混合物递送单链PGY-sRNA-6进入THP-1细胞;
图59a-c:No.41+No.38+鞘氨醇衍生物So-55混合物递送单链PGY-sRNA-6进入THP-1细胞;
图60a-c:No.41+No.38+鞘氨醇衍生物So-56混合物递送单链PGY-sRNA-6进入THP-1细胞;
图61a-c:No.41+No.38+鞘氨醇衍生物So-57混合物递送单链PGY-sRNA-6进入THP-1细胞;
图62a-c:No.41+No.38+鞘氨醇衍生物So-58混合物递送单链PGY-sRNA-6进入THP-1细胞;
图63a-c:No.41+No.38+鞘氨醇衍生物So-59混合物递送单链PGY-sRNA-6进入THP-1细胞;
图64a-c:No.41+No.38+鞘氨醇衍生物So-60混合物递送单链PGY-sRNA-6进入THP-1细胞;
图65a-c:No.41+No.38+鞘氨醇衍生物So-61混合物递送单链PGY-sRNA-6进入THP-1细胞;
图66a-c:No.41+No.38+鞘氨醇衍生物So-62混合物递送单链PGY-sRNA-6进入THP-1细胞;
图67a-c:No.41+No.38+鞘氨醇衍生物So-63混合物递送单链PGY-sRNA-6进入THP-1细胞;
图68a-c:No.41+No.38+鞘氨醇衍生物So-64混合物递送单链PGY-sRNA-6进入THP-1细胞;
图69a-c:No.41+No.38+鞘氨醇衍生物So-65混合物递送单链PGY-sRNA-6进入THP-1细胞;
图70a-c:No.41+No.38+鞘氨醇衍生物So-66混合物递送单链PGY-sRNA-6进入THP-1细胞;
图71a-c:No.41+No.38+鞘氨醇衍生物So-67混合物递送单链PGY-sRNA-6进入THP-1细胞;
图72a-c:No.41+No.38+鞘氨醇衍生物So-68混合物递送单链PGY-sRNA-6进入THP-1细胞;
图73a-c:No.41+No.38+鞘氨醇衍生物So-69混合物递送单链PGY-sRNA-6进入THP-1细胞;
图74a-c:No.41+No.38+鞘氨醇衍生物So-70混合物递送单链PGY-sRNA-6进入THP-1细胞;
图75a-c:No.41+No.38+鞘氨醇衍生物So-71混合物递送单链PGY-sRNA-6进入THP-1细胞;
图76a-c:No.41+No.38+鞘氨醇衍生物So-72混合物递送单链PGY-sRNA-6进入THP-1细胞;
图77a-c:No.41+No.38+鞘氨醇衍生物So-73混合物递送单链PGY-sRNA-6进入THP-1细胞;
图78a-c:No.41+No.38+鞘氨醇衍生物So-74混合物递送单链PGY-sRNA-6进入THP-1细胞;
图79a-c:No.41+No.38+鞘氨醇衍生物So-75混合物递送单链PGY-sRNA-6进入THP-1细胞;
图80a-c:No.41+No.38+No.48+鞘氨醇衍生物So-42混合物递送单链PGY-sRNA-6进入THP-1细胞;
图81a-c:No.41+No.38+No.48+鞘氨醇衍生物So-43混合物递送单链PGY-sRNA-6进入THP-1细胞;
图82a-c:No.41+No.38+No.48+鞘氨醇衍生物So-44混合物递送单链PGY-sRNA-6进入THP-1细胞;
图83a-c:No.41+No.38+No.48+鞘氨醇衍生物So-45混合物递送单链PGY-sRNA-6进入THP-1细胞;
图84a-c:No.41+No.38+No.48+鞘氨醇衍生物So-46混合物递送单链PGY-sRNA-6进入THP-1细胞;
图85a-c:No.41+No.38+No.48+鞘氨醇衍生物So-47混合物递送单链PGY-sRNA-6进入THP-1细胞;
图86a-c:No.41+No.38+No.48+鞘氨醇衍生物So-52混合物递送单链PGY-sRNA-6进入THP-1细胞;
图87a-c:No.41+No.38+No.48+鞘氨醇衍生物So-56混合物递送单链PGY-sRNA-6进入THP-1细胞;
图88a-c:No.41+No.38+No.48+鞘氨醇衍生物So-57混合物递送单链PGY-sRNA-6进入THP-1细胞;
图89a-c:No.41+No.38+No.48+鞘氨醇衍生物So-58混合物递送单链PGY-sRNA-6进入THP-1细胞;
图90a-c:No.41+No.38+No.48+鞘氨醇衍生物So-59混合物递送单链PGY-sRNA-6进入THP-1细胞;
图91a-c:No.41+No.38+No.48+鞘氨醇衍生物So-60混合物递送单链PGY-sRNA-6进入THP-1细胞;
图92a-c:No.41+No.38+No.48+鞘氨醇衍生物So-61混合物递送单链PGY-sRNA-6进入THP-1细胞;
图93a-c:No.41+No.38+No.48+鞘氨醇衍生物So-62混合物递送单链PGY-sRNA-6进入THP-1细胞;
图94a-c:No.41+No.38+No.48+鞘氨醇衍生物So-63混合物递送单链PGY-sRNA-6进入THP-1细胞;
图95a-c:No.41+No.38+No.48+鞘氨醇衍生物So-64混合物递送单链PGY-sRNA-6进入THP-1细胞;
图96a-c:No.41+No.38+No.48+鞘氨醇衍生物So-67混合物递送单链PGY-sRNA-6进入THP-1细胞;
图97a-c:No.41+No.38+No.48+鞘氨醇衍生物So-68混合物递送单链PGY-sRNA-6进入THP-1细胞;
图98a-c:No.41+No.38+No.48+鞘氨醇衍生物So-69混合物递送单链PGY-sRNA-6进入THP-1细胞;
图99a-c:No.41+No.38+No.48+鞘氨醇衍生物So-70混合物递送单链PGY-sRNA-6进入THP-1细胞;
图100:脂质组合递送单链PGY-sRNA-6进入293T的结果;
图101:脂质组合递送单链PGY-sRNA-6进入293T的结果;
图102:脂质组合递送单链PGY-sRNA-6进入293T的结果;
图103:脂质组合递送单链PGY-sRNA-6进入293T的结果;
图104:脂质组合递送单链PGY-sRNA-6进入293T的结果。
具体实施方式
术语定义
本申请所使用的术语可在其前面和/或后面具有单个破折号“-”(或横线)或双重破折号“=”以表明在所提及的取代基和其母体部分之间的键的键级;单个破折号“-”(或横线)表明为单键,双重破折号表明为双键。在没有单个或双重破折号的情况下,理解为在取代基和其母体部分之间形成单键;另外,除非另有说明,意在“从左到右”读取取代基。
不在两个字母或符号之间的短划线(“-”)用于表示取代基的连接点。例如,-C(O)NH2通过碳原子连接。在化学基团的前面或者后面的短划线是为了方便;化学基团可以用或不用一个或多个短划线来描绘,而不会失去其通常的含义。通过结构中的线画出的波浪线指示基团的连接点。
当列举数值的范围时,意图包括在所述范围内的每个数值和子范围。例如“C1-6烷基”意图包括C1、C2、C3、C4、C5、C6、C1-6、C1-5、C1-4、C1-3、C1-2、C2-6、C2-5、C2-4、C2-3、C3-6、C3-5、C3-4、C4-6、C4-5和C5-6烷基。
“烷基”是指含有指定碳原子数的直链或支链的饱和烃链。如本文所述,烷基具有1至24个碳原子(即,C1-24烷基),1至20个碳原子(即,C1-20烷基),1至8个碳原子(即,C1-8烷基),1至6个碳原子(即,C1-6烷基),或1至4个碳原子(即,C1-4烷基)。在一个实施方案中,烷基为C1-6烷基。在一个实施方案中,烷基为C14-20烷基。在一个实施方案中,烷基为直链的C14,C15,C16,C17,C18,C19,C20烷基。
“烯基”是指含有指定碳原子数的且包含至少一个碳-碳双键的脂族烃链。如本文所述,烯基具有2至24个碳原子(即,C2-24烯基),2至20个碳原子(即,C2-20烯基),2至8个碳原子(即,C2-8烯基),2至6个碳原子(即,C2-6烯基),或2至4个碳原子(即,C2-4烯基)。在一个实施方案中,烯基为C14-20烯基。在一个实施方案中,烯基为直链的C14,C15,C16,C17,C18,C19,C20烯基。
如本文所用,术语“酰基”是指基团-CO-。
如本文所用,术语“磷酸酯基”是指基团/>
如本文所用,术语“生物素酰基”是指基团
如本文所用,术语“糖基”是指通过从单糖的环状形式除去半缩醛羟基而获得的一价取代基。例如,术语“1-β-D-葡糖基”是指基团
糖基的实例包括但不限于,葡糖基、乳糖基、半乳糖基、甘露糖基、果糖基和山梨糖基。在一个实施方案中,糖基为β-D-葡糖基。
术语“药学上可接受的盐”是指在合理医学判断的范围内,适于与人类和低等动物的组织接触使用而无不当毒性、刺激、过敏反应等并且与合理的效益/风险比相称的盐。药学上可接受的盐是本领域中熟知的。例如,Berge等人,在J.Pharmaceutical Sciences,1977,66,1-19中详细描述了药学上可接受的盐,通过引用结合在此。本发明化合物的药学上可接受的盐包括衍生自适合的无机和有机酸和碱的盐。药学上可接受的无毒酸加成盐的实例为氨基与无机酸(如盐酸、氢溴酸、磷酸、硫酸以及高氯酸)或与有机酸(如乙酸、草酸、顺丁烯二酸、酒石酸、柠檬酸、琥珀酸或丙二酸)形成的盐,或通过使用本领域中已知的其他方法(例如离子交换法)形成的盐。其他药学上可接受的盐包括己二酸盐、藻酸盐、抗坏血酸盐、天冬氨酸盐、苯磺酸盐、苯甲酸盐、硫酸氢盐、硼酸盐、丁酸盐、樟脑酸盐、樟脑磺酸盐、柠檬酸盐、环戊烷丙酸盐、二葡糖酸盐、十二烷基硫酸盐、乙烷磺酸盐、甲酸盐、富马酸盐、葡庚糖酸盐、甘油磷酸盐、葡糖酸盐、半硫酸盐、庚酸盐、己酸盐、氢碘化物、2-羟基-乙烷磺酸盐、乳糖酸盐、乳酸盐、月桂酸盐、月桂基硫酸盐、苹果酸盐、马来酸盐、丙二酸盐、甲烷磺酸盐、2-萘磺酸盐、烟酸盐、硝酸盐、油酸盐、草酸盐、棕榈酸盐、双羟萘酸盐、果胶酸盐、过硫酸盐、3-苯基丙酸盐、磷酸盐、苦味酸盐、新戊酸盐、丙酸盐、硬脂酸盐、琥珀酸盐、硫酸盐、甲基硫酸盐、酒石酸盐、硫氰酸盐、对-甲苯磺酸盐、十一烷酸盐、戊酸盐等。衍生自适当碱的盐包括碱金属盐、碱土金属盐、铵盐以及N+(C1-4烷基)4 -盐。代表性碱金属或碱土金属盐包括钠盐、锂盐、钾盐、钙盐、镁盐等。适当时,另外的药学上可接受的盐包括使用如卤离子、氢氧根、羧酸根、硫酸根、磷酸根、硝酸根、低级烷基磺酸根以及芳基磺酸根等平衡离子形成的无毒铵、季铵以及胺阳离子。
本申请中所述的逆向蒸发法,是指将核酸的水溶液加入到脂质有机溶剂溶液中,超声,蒸发挥除有机溶剂后,水化得到脂质与核酸的混合物。
本申请所述的水煮法(也称加热法),是指将脂质的有机溶剂溶液加到核酸的水溶液中,约90℃煮15min,得到脂质与核酸的混合物;该方法并不限于水煮升温,还可以通过其它现有技术中已知的升温或加热的方式实现。
逆向蒸发法和水煮法,在受控的温度和混合条件下进行。合适的处理时间、和温度可以由本领域技术人员容易地确定。例如,逆向蒸发法的温度的范围优选约25℃至约70℃,更优选约30℃至约65℃,并且更优选约40℃至约60℃,特别是约55℃。水煮法温度的范围优选约25℃至约100℃,更优选约50℃至约100℃,并且更优选约95℃至约100℃,特别优选为约80℃至100℃。
本申请所述的核酸包括DNA和RNA,优选小RNA,例如所述小RNA长度可以是14-32bp、16-28bp、18-24bp,具体地其长度可以是14、15、16、17、18、19、20、21、22、23、24、25、26、27、28、29、30、31、32bp。
在本申请中,可以通过混合化合物或包含一种或多种所述化合物的组合或组合物和核酸对细胞或受试者递送核酸。在一个实施方案中,化合物具有式I的结构,或其立体异构体或其药学上可接受的盐,
在一个实施方案中,式I的取代基如上文限定。化合物可以是表1中的化合物。包含化合物的组合或组合物可以包含表1中的任一种或多种化合物。本领域技术人员可以根据需要在所述组合或组合物中增加或删除一种或多种化合物。
在一个实施方案中,本申请中的组合或组合物可以是包含No.41、No.38、No.48中任一种或多种的组合。在一个实施方案中,本申请中的组合或组合物可以是包含No.41、No.38、No.48中任一种或多种和选自项11中的任一种或多种化合物的组合。在一个实施方案中,本申请中的组合或组合物可以是包含以下各项的组合:No.41+No.38+鞘氨醇衍生物So-1;No.41+No.38+鞘氨醇衍生物So-2;No.41+No.38+鞘氨醇衍生物So-3;No.41+No.38+鞘氨醇衍生物So-4;No.41+No.38+鞘氨醇衍生物So-5;No.41+No.38+鞘氨醇衍生物So-6;No.41+No.38+鞘氨醇衍生物So-7;No.41+No.38+鞘氨醇衍生物So-8;No.41+No.38+鞘氨醇衍生物So-9;No.41+No.38+鞘氨醇衍生物So-10;No.41+No.38+鞘氨醇衍生物So-11;No.41+No.38+鞘氨醇衍生物So-12;No.41+No.38+鞘氨醇衍生物So-13;No.41+No.38+鞘氨醇衍生物So-14;No.41+No.38+鞘氨醇衍生物So-15;No.41+No.38+鞘氨醇衍生物So-45;No.41+No.38+鞘氨醇衍生物So-46;No.41+No.38+鞘氨醇衍生物So-47;No.41+No.38+鞘氨醇衍生物So-48;No.41+No.38+鞘氨醇衍生物So-49;No.41+No.38+鞘氨醇衍生物So-50;No.41+No.38+鞘氨醇衍生物So-51;No.41+No.38+鞘氨醇衍生物So-52;No.41+No.38+鞘氨醇衍生物So-53;No.41+No.38+鞘氨醇衍生物So-54;No.41+No.38+鞘氨醇衍生物So-55;No.41+No.38+鞘氨醇衍生物So-56;No.41+No.38+鞘氨醇衍生物So-57;No.41+No.38+鞘氨醇衍生物So-58;No.41+No.38+鞘氨醇衍生物So-59;No.41+No.38+鞘氨醇衍生物So-60;No.41+No.38+鞘氨醇衍生物So-61;No.41+No.38+鞘氨醇衍生物So-62;No.41+No.38+鞘氨醇衍生物So-63;No.41+No.38+鞘氨醇衍生物So-64;No.41+No.38+鞘氨醇衍生物So-65;No.41+No.38+鞘氨醇衍生物So-66;No.41+No.38+鞘氨醇衍生物So-67;No.41+No.38+鞘氨醇衍生物So-68;No.41+No.38+鞘氨醇衍生物So-69;No.41+No.38+鞘氨醇衍生物So-70;No.41+No.38+鞘氨醇衍生物So-71;No.41+No.38+鞘氨醇衍生物So-72;No.41+No.38+鞘氨醇衍生物So-73;No.41+No.38+鞘氨醇衍生物So-74;No.41+No.38+鞘氨醇衍生物So-75;No.41+No.38+No.48+鞘氨醇衍生物So-42;No.41+No.38+No.48+鞘氨醇衍生物So-43;No.41+No.38+No.48+鞘氨醇衍生物So-44;No.41+No.38+No.48+鞘氨醇衍生物So-45;No.41+No.38+No.48+鞘氨醇衍生物So-46;No.41+No.38+No.48+鞘氨醇衍生物So-47;No.41+No.38+No.48+鞘氨醇衍生物So-52;No.41+No.38+No.48+鞘氨醇衍生物So-56;No.41+No.38+No.48+鞘氨醇衍生物So-57;No.41+No.38+No.48+鞘氨醇衍生物So-58;No.41+No.38+No.48+鞘氨醇衍生物So-59;No.41+No.38+No.48+鞘氨醇衍生物So-60;No.41+No.38+No.48+鞘氨醇衍生物So-61;No.41+No.38+No.48+鞘氨醇衍生物So-62;No.41+No.38+No.48+鞘氨醇衍生物So-63;No.41+No.38+No.48+鞘氨醇衍生物So-64;No.41+No.38+No.48+鞘氨醇衍生物So-67;No.41+No.38+No.48+鞘氨醇衍生物So-68;No.41+No.38+No.48+鞘氨醇衍生物So-69;No.41+No.38+No.48+鞘氨醇衍生物So-70;No.41+No.38+鞘氨醇衍生物So-23、25、26、27、29、30、31、32、33、34、35、36、37、38、40、41、42、43、44、70、71或73中任一种或多种。本领域技术人员可以选择适当的浓度和使用体积。优选地,化合物的使用浓度如表1所述。各种化合物的比例为0.1-10∶0.1-10,0.2-9∶0.2-9,0.3-8∶0.3-8;0.4-7∶0.4-7;0.5-6∶0.5-6;0.6-5∶0.6-5;0.7-4∶0.7-4;0.8-3∶0.8-3;0.9-2∶0.9-2;更优选1∶1,或者它们之间的任何比例。本领域技术人员可以根据化合物的母液浓度,适当调整各种化合物的比例。此外,本申请证明了特定的脂质组合可以有效促进核酸的递送,效果优于单一脂质的效果简单叠加。
在一个实施方案中,核酸是合成或纯化的、治疗性或非治疗性、和/或诊断性或非诊断性的,例如选自RNA和DNA,例如选自单链或双链或部分双链的RNA和DNA。在核酸是治疗性或诊断性时,所述核酸可以用于治疗或诊断选自下组的疾病:炎性疾病、肺炎、心肌炎、急慢性胃炎、急慢性肠炎、急慢性肝炎、急慢性肾炎、皮炎、脑炎、淋巴炎、结膜炎、角膜炎、虹膜睫状体炎、中耳炎、过敏性鼻炎、哮喘、肺纤维化慢性阻塞性肺疾病、过敏性皮炎、镰状细胞病、多发性硬化、***性红斑狼疮、狼疮性肾炎、肺癌、胃癌、结直肠癌、肝癌、胰腺癌、***、乳腺癌、白血病、多发性骨髓瘤、糖尿病和痛风。本领域技术人员可以根据具体情况选择合适的核酸。例如,RNA可以是信使RNA(mRNA)、rRNA(核糖体RNA)、tRNA(转运RNA)、不均一核RNA(hnRNA)、小核RNA(snRNA)、核仁小RNA(snoRNA)、小胞质RNA、小RNA、转移-信使RNA(tmRNA)、端粒酶RNA和反义RNA,优选小RNA。
在一个实施方案中,递送包括通过加热法、逆向蒸发法、直接混合、反复冻融和/或薄膜分散处理化合物、其立体异构体或其药学上可接受的盐或包含它们的组合。本领域技术人员可以选择合适的方式来进行递送。在一个实施方案中,加热法的温度为约0℃至约100℃,约25℃至约100℃,优选约80℃至约100℃,例如4℃,37℃,60℃,80℃或100℃;所述逆向蒸发法的制备温度为约25℃至约70℃,优选约55℃,加热时间为约0分钟-约24小时,约5分钟-约20小时,约5分钟-约16小时,约5分钟-约10小时,约5分钟-约4小时,或者约10小时-约1小时,优选15分钟。
在本申请中,通过化合物对核酸进行处理,可以通过口服直接对受试者进行施用处理后的混合物。此外,也可以通过其他方式对受试者进行施用,例如可以静脉内施用,如推注或灌注,通过皮下、肌肉内、动脉内、腹膜内、肺内、脑脊髓内、关节内、滑膜内、鞘内、损伤内、和/或吸入路径如鼻内,通常通过静脉内或皮下施用。
本发明还提供了化合物组合、组合物或试剂盒,它们包含本申请中所述的任何化合物,例如表1中的任一种或多种化合物。本领域技术人员可以根据需要在所述化合物组合、组合物或试剂盒中增加其他化合物,只要它们保留递送核酸的功能。
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表2.小RNA合成信息(购自广州锐博生物科技有限公司,Ribobio)
名称 | 单/双链 | 序列 | 浓度 |
PGY-ssRNA-6 | 单链 | GTTCAGAGTTCTACAGTCCGA | 20μM |
表3.总体系2升的PBS配方(试剂购自北京化工厂)
PBS总体系 | 2升 |
NaCl | 16g |
Na2HPO4·12H2O | 7.16g |
KH2PO4 | 0.48g |
KCl | 0.4g |
实施例
实验方法
1.细胞的培养:
实验所用人胚肾细胞系HEK293T,人单核细胞THP-1购自北京协和医学院细胞培养中心。细胞均置于37℃、5%CO2培养箱内培养。其中,HEK293T细胞培养于DMEM培养基(HyClone)培养;THP-1细胞培养于RPMI-1640培养基(HyClone)培养;各培养基均含10%胎牛血清和一定比例的抗生素(青霉素100U/ml&链霉素100mg/ml)。细胞培养至对数生长期,细胞密度为6×105/1mL培养基/孔;分至12孔板(1ml培养基/孔),37℃孵育过夜(12h)后进行后续实验。
2.鞘氨醇类脂质递送小RNA体系的制备
2.1向1.5ml EP管中加入5μl小RNA(Ribobio,20μM,如表2所示)及95μL DEPC(Sigma)处理过的水,混匀后加入一定量脂质单体或脂质组合物氯仿溶液(如表1所示),充分吹吸混匀;
2.2将体系充分混匀后于90℃水浴加热15分钟;
2.3把体系取出冷却至室温,即得。
3.实时荧光定量PCR(RT-qPCR)检测细胞内脂质递送核酸的表达量
3.1实验所用人胚肾细胞系HEK293T培养至对数生长期,然后分别铺板到12孔板,细胞密度为6×105/1mL培养基/孔;12孔板(1mL培养基/孔),37℃孵育过夜(12h)后进行后续实验。
3.2实验分组如下:
1)空白组:是指未经处理的细胞,该组作为空白对照组。
2)自由摄取组:直接加入5μL小RNA溶液(贮存浓度20μM),该组作为阴性对照组;
3)脂质核酸混合物处理组:将步骤2中制备的脂质与小RNA混合物加入细胞中,混匀,小RNA的终浓度为100nM。
3.3与细胞共同孵育特定时间后,用PBS清洗细胞3次,用TRIzol裂解液(购自Sigma-Aldrich)收取细胞,提取其中总RNA,利用RT-qPCR(SYBR Green染料法)检测进入细胞的小RNA丰度,具体步骤如下:
1)提取细胞总RNA:
12孔板培养的细胞(约1×106个细胞/孔),每孔加入1mL TRIzol裂解液后,先置于冰上,待所有的样品都加入TRIzol后,室温放置5min,使其充***解;
4℃,12,000rpm离心5min,弃沉淀,将TRIzol转移到新的离心管中;
按200μL氯仿/mL TRIzol加入氯仿,充分振荡,混匀后室温放置5min;
4℃,12,000rpm,离心15min;
吸取上层水相,至另一离心管中,按0.5mL异丙醇/mL TRIzol加入异丙醇混匀,室温放置5-10min;
4℃,12,000rpm,离心15min,弃上清,RNA沉于管底;
加入1mL 75%乙醇,温和振荡离心管,悬浮沉淀;
4℃,12,000rpm,离心10min,弃上清,加入1mL 75%乙醇,温和振荡离心管,悬浮沉淀;
4℃,12,000rpm,离心10min,弃上清,室温晾干,用50μL RNase-free的H2O溶解RNA样品,测OD值定量RNA浓度。
2)将总RNA逆转录为cDNA:通过逆转录试剂盒(High-Capacity cDNA ReverseTranscription Kits,Applied Biosystems,cat.no.4368813),用茎环法(stem-loop法)(参见例如Real-time quantification of microRNAs by stem-loop RT-PCR,NucleicAcids Res.2005 Nov 27;33(20):e179,通过引用并入本文)将sRNA逆转录为cDNA,逆转录体系如下:模板RNA(150ng/μL)10μL,10X RT缓冲液2.0μL,25X dNTP Mix(100mM)0.8μL,U6RT stem-loop引物2.0μL,PGY-sRNA-6 RT stem-loop引物2.0μL,MultiScribeTM逆转录酶1.0μL,RNA酶抑制剂1.0μL,无核酸酶H2O1.2μL,瞬时离心后,放入PCR仪反应,反应条件如下:(1)25℃,10min;(2)37℃,120min;(3)85℃,5min;(4)4℃,终止反应。反应结束后加入20μL无RNA酶ddH2O,补足终体积至40μL。该逆转录过程中使用的茎环法引物由北京擎科新业生物技术有限公司合成(U6 RT引物,因为RT-qPCR反应对小RNA的定量只能是相对定量,所以以U6作为标准的参考基因,计算其相对表达量):U6 RT stem-loop引物:GTCGTATCCAGTGCAGGGTCCGAGGTATTCGCACTGGATACGACAAAAATATG;PGY-sRNA-6 RT stem-loop引物:GTCGTATCCAGTGCACGCTCCGAGGTATTCGCACTGGATACGACTCGGAC)。
3)定量PCR扩增反应:qPCR反应体系总体积10μL,包括:5μL2×SYBR Green MasterMix,0.5μL正向引物(10μM),0.5μL反向引物(10μM),1μL逆转录得到的cDNA,3μL无RNA酶H2O。使用LightCycler 480荧光定量PCR仪,PCR反应条件是:95℃,持续5min预变性,开始进入PCR扩增循环:(1)95℃,10s;(2)55℃,10s;(3)72℃,20s;总共进行40个循环;最后40℃持续10s降温。扩增反应正向引物和反向引物均由北京擎科新业生物技术有限公司设计和合成(U6正向引物:GCGCGTCGTGAAGCGTTC,U6反向引物:GTGCAGGGTCCGAGGT,PGY-sRNA-6正向引物:TCGCGCGTTCAGAGTTC,反向引物miR-all Reverse primer:GTGCACGCTCCGAGGT)。
4)利用2-ΔCt法(基因相对表达量=2-(Ct目的基因-Ct内参基因))计算相对进入量(单链或双链RNA)。
4.流式细胞技术(CFlow)检测细胞内脂质递送核酸的细胞摄取量
4.1主要实验仪器及器材:
10cm细胞培养皿、12孔细胞培养板、移液器、移液管、光学显微镜、流式细胞仪C6仪器(购自美国BD公司);
4.2主要实验试剂:
模型建立及转染:表1中所示的人工合成脂质单体、表2中所示的人工合成5’-FAM标记的sRNA(单链,Ribobio);
4.3实验所用人胚肾细胞系HEK293T,人单核细胞THP-1细胞培养至对数生长期,然后分别铺板到12孔板,细胞密度为6×105/1mL培养基/孔;12孔板(1mL培养基/孔),37℃孵育过夜(12h)后进行后续实验。
4.4实验分组如下:
1)空白组:是指未经处理的细胞,该组作为空白对照组。
2)自由摄取组:直接加入5mL 5’-FAM标记的小RNA溶液(贮存浓度为20μM),该组作为阴性对照组;
3)脂质核酸混合物处理组:将步骤2中制备的脂质与5’-FAM标记的小RNA混合物加入细胞中,混匀,小RNA的终浓度为100nM。
4.5与细胞共同孵育6h/9h后,用PBS清洗细胞3次,用PBS(自配)重悬收取细胞,使用流式细胞仪C6仪器(购自美国BD公司)检测样品孔细胞的荧光强度。实施方法:打开流式细胞仪/>C6仪器检测用软件CFlow Plus,用清洁液清洗仪器3分钟,再用双蒸水清洗5分钟;将空白组样品开始进样并设置不限定细胞数量,当检测至细胞数量为10000events时停止,并图出荧光图谱(横坐标:FSC-A,纵坐标:SSC-A)中的活细胞类群作为门P1;限定细胞数量,检测至门P1内细胞数量为10000events时停止,在荧光图谱(横坐标:荧光通道FLA-1,纵坐标:SSC-A)中获取荧光偏移值。
实施例1:不同浓度鞘氨醇衍生物单体递送小RNA进入HEK293T细胞。
(一)实验分组:
A:空白组:指未经处理过的细胞,该组作为空白对照;
B:自由摄取组:用鞘氨醇的溶剂CHCl3作为载体递送FAM标记的小RNA,核酸终浓度100nM,该组作为阴性对照组;
C:3.75ug表4中所示的鞘氨醇衍生物单体溶液递送0.1nmol FAM标记的单链PGY-sRNA-6进入细胞,核酸终浓度100nM;
D:12.5ug鞘氨醇衍生物单体溶液递送0.1nmol FAM标记的单链PGY-sRNA-6进入细胞,核酸终浓度100nM;
E:37.5ug鞘氨醇衍生物单体溶液递送0.1nmol FAM标记的单链PGY-sRNA-6进入细胞,核酸终浓度100nM;
(二)实验过程
1)水煮法条件:100μL FAM标记的单链PGY-sRNA-6溶液中加入相应量脂质溶液,90℃,加热15min;
2)实验条件:小RNA终浓度100nM,加入细胞9h后,经流式细胞术检测荧光偏移比较细胞中单链PGY-sRNA-6进入量。具体实验方法参见上文“流式细胞技术(CFlow)检测细胞内脂质递送核酸的细胞摄取量”。所有实验一式三份进行。
结论:结果表明,与自由摄取组相比,实验组中鞘氨醇脂质递送荧光标记的核酸后荧光值有明显偏移,表明不同的鞘氨醇脂质衍生物单体均可以有效递送单链小RNA进入细胞(图1a-e至图33a-e)。鞘氨醇脂质衍生物单体对核酸的递送有效性,鞘氨醇衍生物递送sRNA的效率对脂质呈浓度依赖性,本文中高浓度鞘氨醇衍生物递送(37.5ug鞘氨醇衍生物溶液递送0.1nmol的小RNA)具有更加明显的效果(见表4)。
实施例2:不同鞘氨醇衍生物、鞘氨醇(So)和磷脂酰乙醇胺(PE)混合物递送单链PGY-sRNA-6进入THP-1细胞。
(一)实验分组:
A:空白组:指未经处理过的细胞,该组作为空白对照;
B:自由摄取组:用鞘氨醇的溶剂CHCl3作为载体递送FAM标记的小RNA,核酸终浓度100nM,该组作为阴性对照组;
C:脂质混合物递送组:12.5μg鞘氨醇衍生物的脂质混合物(No.41∶No.38∶So=1∶1∶1,其中具体的脂质混合物如图34a-c-图79a-c,表4中所示)递送0.1nmol FAM标记的单链PGY-sRNA-6进入细胞,核酸终浓度100nM;
(二)实验过程
1)水煮法条件:100μL FAM标记的小RNA溶液中加入12.5μg脂质混合物溶液,90℃,加热15min;
2)实验条件:小RNA终浓度100nM,加入细胞6h后,经流式细胞术检测荧光偏移比较细胞中小RNA的进入量。具体实验方法参见上文“流式细胞技术(CFlow)检测细胞内脂质递送核酸的细胞摄取量”。所有实验一式三份进行。
结论:结果表明,与自由摄取组相比,实验组中鞘氨醇脂质递送核酸后荧光值有明显偏移(图34a-c-图79a-c,表4),表明12.5μg脂质混合物(No.41∶No.38∶So=1∶1∶1)可以有效递送小RNA进入细胞,脂质混合物(No.41∶No.38∶So=1∶1∶1)对核酸的递送有效性(见表4),脂质混合物递送小RNA有希望提高临床上核酸药物递送的效率。
实施例3.不同鞘氨醇衍生物、鞘氨醇(So)磷脂酰乙醇胺(PE)和甘油三酯(TG)混合物递送单链PGY-sRNA-6进入THP-1细胞。
(一)实验分组:
A:空白组:指未经处理过的细胞,该组作为空白对照;
B:自由摄取组:用鞘氨醇的溶剂CHCl3作为载体递送FAM标记的小RNA,核酸终浓度100nM,该组作为阴性对照组;
C:脂质混合物递送组:12.5μg鞘氨醇衍生物的脂质混合物(No.41∶No.38∶No.48∶So=2∶2∶1∶2,其中具体的脂质混合物如图80a-c-图99a-c,表4中所示)递送0.1nmol FAM标记的单链PGY-sRNA-6进入细胞,核酸终浓度100nM;
(二)实验过程
1)水煮法条件:100μL FAM标记的小RNA溶液中加入12.5μg脂质混合物溶液,90℃,加热15min;
2)实验条件:小RNA终浓度100nM,加入细胞6h后,经流式细胞术检测荧光偏移比较细胞中小RNA的进入量。具体实验方法参见上文“流式细胞技术(CFlow)检测细胞内脂质递送核酸的细胞摄取量”。所有实验一式三份进行。
结论:结果表明,与自由摄取组相比,实验组中鞘氨醇脂质递送核酸后荧光值有明显偏移(图80a-c-图99a-c,表4),表明12.5μg脂质混合物(No.41∶No.38∶No.48∶So=2∶2∶1∶2)可以有效递送小RNA进入细胞,脂质混合物(No.41∶No.38∶No.48∶So=2∶2∶1∶2)对核酸的递送有效性(见表4),脂质混合物递送小RNA有希望提高临床上核酸药物递送的效率。
实施例4:不同鞘氨醇衍生物、鞘氨醇(So)和磷脂酰乙醇胺(PE)混合物递送单链PGY-sRNA-6进入HEK293T细胞。
(一)实验分组:
A:空白组:指未经处理过的细胞,该组作为空白对照;
B:自由摄取组:用鞘氨醇的溶剂CHCl3作为载体递送小RNA,核酸终浓度100nM,该组作为阴性对照组;
C:脂质混合物递送组:12.5μg鞘氨醇衍生物的脂质混合物(No.41∶No.38∶So=1∶1∶1,其中具体的脂质混合物如图100-104中所示)递送0.1nmol的单链PGY-sRNA-6进入细胞,核酸终浓度100nM;
(二)实验过程
1)水煮法条件:100μL小RNA溶液中加入相应量脂质溶液,90℃,加热15min;
2)实验条件:小RNA终浓度100nM,加入细胞6h后,经RT-qPCR检测比较细胞中小RNA的进入量。具体实验方法参见上文“实时荧光定量PCR(RT-qPCR)检测细胞内脂质递送核酸的表达量”。所有实验一式三份进行。
结论:结果表明,与空白组及阴性对照组相比,实验组中含鞘氨醇衍生物的脂质组合(No.41∶No.38∶So=1∶1∶1)12.5ug递送0.1nmol核酸后检测到细胞的小RNA相对摄取量有明显增加,表明多种鞘氨醇脂质单体衍生物的加入可以极大程度提高脂质组合的递送效率(见图100-104)。
表4
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说明:1.实验组与阴性对照组相比,荧光偏移(向右偏移,以红色区域标记)量增加,表示细胞摄入荧光标记的RNA量更多,即脂质递送入细胞的RNA量更多,即反映递送效率更高;
2.递送效率随脂质浓度改变说明,RNA的递送基于脂质浓度呈依赖性。
Claims (29)
1.脂质组合物在制备对细胞或受试者递送核酸的脂质-核酸混合物中的应用,所述制备包括:
将脂质组合物与所述核酸混合得到脂质-核酸混合物;
其中所述脂质组合物由一种或多种式Ia化合物、其立体异构体或其药学上可接受的盐组成:
其中:
R1为含有一个双键的C15烯基;
R2为氢,且R3为羟基;
R4为氢;
R5选自氢或C14烷基酰基;
R6为葡糖基或者半乳糖基。
2.根据权利要求1所述的应用,其中所述脂质组合物包含如下脂质中的一种或多种:
3.根据权利要求1所述的应用,其中所述核酸是合成或纯化的、治疗性或非治疗性、和/或诊断性或非诊断性的;
其中在核酸是治疗性或诊断性时,所述核酸用于治疗或诊断选自下组的疾病:炎性疾病、哮喘、肺纤维化慢性阻塞性肺疾病、镰状细胞病、多发性硬化、***性红斑狼疮、肺癌、胃癌、结直肠癌、肝癌、胰腺癌、***、乳腺癌、白血病、多发性骨髓瘤、糖尿病和痛风。
4.根据权利要求3所述的应用,其中所述核酸选自RNA和DNA。
5.根据权利要求4所述的应用,其中所述核酸选自单链或双链或部分双链的RNA和DNA。
6.根据权利要求4所述的应用,其中所述RNA选自:信使RNA(mRNA)、rRNA(核糖体RNA)、tRNA(转运RNA)、不均一核RNA(hnRNA)、小核RNA(snRNA)、核仁小RNA(snoRNA)、小胞质RNA、小RNA、转移-信使RNA(tmRNA)、端粒酶RNA和反义RNA。
7.根据权利要求6所述的应用,其中所述RNA为小RNA。
8.根据权利要求7所述的应用,其中所述小RNA长度是14-32bp。
9.根据权利要求7所述的应用,其中所述小RNA长度是16-28bp。
10.根据权利要求7所述的应用,其中所述小RNA长度是18-24bp。
11.根据权利要求8所述的应用,其中所述小RNA包含GTTCAGAGTTCTACAGTCCGA的序列。
12.根据权利要求3所述的应用,其中所述炎性疾病选自下组的疾病:肺炎、心肌炎、急慢性胃炎、急慢性肠炎、急慢性肝炎、急慢性肾炎、皮炎、脑炎、淋巴炎、结膜炎、角膜炎、虹膜睫状体炎、中耳炎、过敏性鼻炎、过敏性皮炎和狼疮性肾炎。
13.根据权利要求1所述的应用,其中通过加热法、逆向蒸发法、直接混合、反复冻融和/或薄膜分散将所述脂质组合物与所述核酸混合得到所述脂质-核酸混合物。
14.根据权利要求13所述的应用,其中
所述加热法包括将包含所述脂质组合物的有机溶剂溶液加入到包含核酸的水溶液中,随后在25℃至100℃的温度下加热,从而获得所述脂质-核酸混合物;
所述逆向蒸发法包括将核酸的水溶液加入到所述脂质组合物的有机溶剂溶液中,超声,蒸发挥除有机溶剂后进行水化,从而获得所述脂质-核酸混合物,其中所述逆向蒸发法的制备温度为25℃至70℃,加热时间为0分钟-24小时。
15.根据权利要求14所述的应用,其中所述加热法包括将包含所述脂质组合物的有机溶剂溶液加入到包含核酸的水溶液中,随后在50℃至100℃的温度下加热,从而获得所述脂质-核酸混合物。
16.根据权利要求14所述的应用,其中所述加热法包括将包含所述脂质组合物的有机溶剂溶液加入到包含核酸的水溶液中,随后在80℃至100℃的温度下加热,从而获得所述脂质-核酸混合物。
17.根据权利要求14所述的应用,其中所述加热法包括将包含所述脂质组合物的有机溶剂溶液加入到包含核酸的水溶液中,随后在95℃至100℃的温度下加热,从而获得所述脂质-核酸混合物。
18.根据权利要求14所述的应用,其中所述逆向蒸发法的制备温度为55℃。
19.根据权利要求14所述的应用,其中所述逆向蒸发法的加热时间为5分钟-20小时。
20.根据权利要求14所述的应用,其中所述逆向蒸发法的加热时间为5分钟-16小时。
21.根据权利要求14所述的应用,其中所述逆向蒸发法的加热时间为5分钟-10小时。
22.根据权利要求14所述的应用,其中所述逆向蒸发法的加热时间为5分钟-4小时。
23.根据权利要求14所述的应用,其中所述逆向蒸发法的加热时间为10分钟-1小时。
24.根据权利要求14所述的应用,其中所述逆向蒸发法的加热时间为15分钟。
25.根据权利要求1所述的应用,其中将所述核酸体外递送至细胞,或体内递送至受试者。
26.根据权利要求25所述的应用,其中体内递送至受试者包括口服、静脉内施用,通过皮下、肌肉内、动脉内、腹膜内、肺内、脑脊髓内、关节内、滑膜内、鞘内、损伤内、和/或吸入路径施用。
27.根据权利要求26所述的应用,其中体内递送至受试者包括静脉内或皮下施用。
28.根据权利要求26所述的应用,其中体内递送至受试者包括推注或灌注。
29.根据权利要求26所述的应用,其中所述吸入路径为鼻内。
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Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994023694A1 (en) * | 1993-04-20 | 1994-10-27 | Unilever Plc | Cosmetic composition containing ceramide precursors |
WO1997019184A1 (en) * | 1995-11-21 | 1997-05-29 | Paavo Kinnunen | Liposomal transfection method |
WO2005014008A2 (en) * | 2003-07-17 | 2005-02-17 | The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Treatment of disorders associated with natural killer t cells |
CN101939027A (zh) * | 2007-10-02 | 2011-01-05 | Mdrna有限公司 | 用于递送核酸的脂肽 |
CN105078889A (zh) * | 2015-07-15 | 2015-11-25 | 魏垒 | 一种用于治疗软骨疾病的脂质体递送***及其制备方法 |
WO2017053681A1 (en) * | 2015-09-24 | 2017-03-30 | Wisconsin Alumni Research Foundation | Methods of expanding hematopoietic stem cells, compositions, and methods of use thereof |
WO2018177383A1 (zh) * | 2017-03-29 | 2018-10-04 | 中国医学科学院基础医学研究所 | 化合物或中药提取物在制备核酸递送试剂中的应用及其相关产品 |
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KR101306097B1 (ko) * | 2010-10-06 | 2013-09-09 | 성균관대학교산학협력단 | 피에이치 민감성 폴리머좀의 제조방법 |
-
2019
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Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1994023694A1 (en) * | 1993-04-20 | 1994-10-27 | Unilever Plc | Cosmetic composition containing ceramide precursors |
WO1997019184A1 (en) * | 1995-11-21 | 1997-05-29 | Paavo Kinnunen | Liposomal transfection method |
WO2005014008A2 (en) * | 2003-07-17 | 2005-02-17 | The Government Of The United States Of America, As Represented By The Secretary, Department Of Health And Human Services | Treatment of disorders associated with natural killer t cells |
CN101939027A (zh) * | 2007-10-02 | 2011-01-05 | Mdrna有限公司 | 用于递送核酸的脂肽 |
CN105078889A (zh) * | 2015-07-15 | 2015-11-25 | 魏垒 | 一种用于治疗软骨疾病的脂质体递送***及其制备方法 |
WO2017053681A1 (en) * | 2015-09-24 | 2017-03-30 | Wisconsin Alumni Research Foundation | Methods of expanding hematopoietic stem cells, compositions, and methods of use thereof |
WO2018177383A1 (zh) * | 2017-03-29 | 2018-10-04 | 中国医学科学院基础医学研究所 | 化合物或中药提取物在制备核酸递送试剂中的应用及其相关产品 |
Non-Patent Citations (1)
Title |
---|
Novel cationic liposomes for DNA-transfection with high efficiency and low toxicity;Tommi Paukku,et al.;《Chemistry and Physics of Lipids》;19971231;第87卷;23-29 * |
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