CN1355713A - 可释放的键和含有该键的组合物 - Google Patents
可释放的键和含有该键的组合物 Download PDFInfo
- Publication number
- CN1355713A CN1355713A CN00807707A CN00807707A CN1355713A CN 1355713 A CN1355713 A CN 1355713A CN 00807707 A CN00807707 A CN 00807707A CN 00807707 A CN00807707 A CN 00807707A CN 1355713 A CN1355713 A CN 1355713A
- Authority
- CN
- China
- Prior art keywords
- poly
- liposome
- chemical compound
- compositions
- mpeg
- Prior art date
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- Granted
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- 239000000203 mixture Substances 0.000 title claims description 62
- 150000001875 compounds Chemical class 0.000 claims abstract description 82
- 150000001412 amines Chemical class 0.000 claims abstract description 50
- 239000003446 ligand Substances 0.000 claims abstract description 38
- 229920001477 hydrophilic polymer Polymers 0.000 claims abstract description 29
- 125000004396 dithiobenzyl group Chemical group 0.000 claims abstract description 22
- 239000002502 liposome Substances 0.000 claims description 122
- -1 amine lipid Chemical class 0.000 claims description 65
- 229920001223 polyethylene glycol Polymers 0.000 claims description 61
- 238000006243 chemical reaction Methods 0.000 claims description 57
- 239000002202 Polyethylene glycol Substances 0.000 claims description 56
- 229920001184 polypeptide Polymers 0.000 claims description 53
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 53
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 52
- 239000003814 drug Substances 0.000 claims description 35
- 125000000217 alkyl group Chemical group 0.000 claims description 32
- 125000003118 aryl group Chemical group 0.000 claims description 26
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- 238000000034 method Methods 0.000 claims description 13
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- NYEZZYQZRQDLEH-UHFFFAOYSA-N 2-ethyl-4,5-dihydro-1,3-oxazole Chemical compound CCC1=NCCO1 NYEZZYQZRQDLEH-UHFFFAOYSA-N 0.000 claims description 6
- GNSFRPWPOGYVLO-UHFFFAOYSA-N 3-hydroxypropyl 2-methylprop-2-enoate Chemical compound CC(=C)C(=O)OCCCO GNSFRPWPOGYVLO-UHFFFAOYSA-N 0.000 claims description 6
- YSFGBPCBPNVLOK-UHFFFAOYSA-N 6-hydroxy-2-methylhex-2-enamide Chemical compound NC(=O)C(C)=CCCCO YSFGBPCBPNVLOK-UHFFFAOYSA-N 0.000 claims description 6
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- DGIBHCWBCOAPDN-UHFFFAOYSA-N 1-hydroxy-6-(trifluoromethyl)benzotriazole Chemical compound C1=C(C(F)(F)F)C=C2N(O)N=NC2=C1 DGIBHCWBCOAPDN-UHFFFAOYSA-N 0.000 claims description 3
- GTFDJMHTJNPQFS-UHFFFAOYSA-N 1-hydroxypiperidine-2,6-dione Chemical compound ON1C(=O)CCCC1=O GTFDJMHTJNPQFS-UHFFFAOYSA-N 0.000 claims description 3
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- FZZPTFZNSPIAGO-UHFFFAOYSA-N 2-hydroxy-3a,4,5,7a-tetrahydroisoindole-1,3-dione Chemical compound C1CC=CC2C(=O)N(O)C(=O)C21 FZZPTFZNSPIAGO-UHFFFAOYSA-N 0.000 claims description 3
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- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 claims description 3
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- VTGOHKSTWXHQJK-UHFFFAOYSA-N pyrimidin-2-ol Chemical compound OC1=NC=CC=N1 VTGOHKSTWXHQJK-UHFFFAOYSA-N 0.000 claims description 3
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- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 50
- 235000018417 cysteine Nutrition 0.000 description 50
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- 239000002904 solvent Substances 0.000 description 24
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- 230000009182 swimming Effects 0.000 description 23
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Abstract
描述了一种含有通过二硫代苄基键共价但可逆地连接到含胺配体上的亲水性聚合物的化合物。
Description
发明领域
本发明涉及一种化合物,该化合物含有可断裂地连接到含胺配体上的亲水性聚合物如聚乙二醇,在优选实施方案中该含胺配体可以是含胺脂质、药物或蛋白质。在温和的硫解条件下该化合物能够离解,再生成其初始形式的含胺配体。
发明背景
亲水性聚合物,如聚乙二醇(PEG),已用于修饰各种底物如多肽、药物和脂质体,以降低底物的免疫原性和/或改善其血液循环寿命。
例如,蛋白质肠胃外给药可以是致免疫的,并可能具有短药理学半衰期。而且蛋白质是较不溶于水的。所以,要使蛋白质在病人体内达到治疗有效量的血液水平是困难的。将PEG缀合到蛋白质上被认为是克服这些困难的一种途径。Davis等在美国专利4,179,337中公开了将PEG缀合到蛋白质如酶和胰岛素以形成具有较低免疫原性但仍保持相当程度的生理活性的PEG-蛋白质缀合物。Veronese等(Applied Biochem.和Biotech,11:141-152(1985))公开了用氯甲酸苯酯活化聚乙二醇以修饰核糖核酸酶和超氧化物歧化酶。Katre等人在美国专利号4,766,106和4,917,888中公开了通过聚合物缀合而增溶蛋白质。PEG和其它聚合物缀合到重组蛋白质上以降低免疫原性和增大半衰期(Nitecki等人,美国专利号4,902,502;Enzon,Inc.,PCT/US90/02133)。Garman(美国专利号4,935,465)描述了用水溶性聚合物修饰的蛋白质,该聚合物通过一种可逆键合基团结合到该蛋白质上。
但是,至今所述的PEG-蛋白质缀合物受损于许多缺陷。例如,用PEG修饰蛋白质经常钝化蛋白质从而使所得缀合物具有不良的生物活性。在现有技术中一般期望具有稳定结合到该蛋白质上的PEG以保持PEG提供的有益性能。某些蛋白质-PEG缀合物的另一问题在于分解该缀合物时可能会产生不良的产物。
PEG还被描述用于改善脂质体的血液循环寿命(美国专利号5,103,556)。这里的PEG共价结合到该脂质的极性头基团上以掩蔽或屏蔽脂质体免受识别和通过网状内皮系统被除去。还描述了具有可释放的PEG链的脂质体,其中PEG链在受到适宜的刺激如pH的变化时从该脂质体中释放出来(PCT/US97/18813)。但是,从该脂质体中释放出PEG链受损于该缺陷,即分解产物是化学修饰的并可能具有不可预知的潜在的体内负作用。
发明简述
因此,本发明的一个目的在于提供一种化合物,其中一配体共价而可逆地连接到亲水性聚合物上。在断裂该键时再生其天然形式的配体。
其中R1为含有用于连接到二硫代苄基部分的键的亲水性聚合物;R2选自H、烷基和芳基;R3选自O(C=O)R4、S(C=O)R4和O(C=S)R4;R4包括含胺配体;而R5选自H、烷基和芳基;其中CH2-R3的取向选自邻位和对位。
在一个实施方案中,R5为H而R2选自CH3、C2H5和C3H8。在另一实施方案中,R2和R5为烷基。
在另一实施方案中,含胺配体R4选自多肽、含胺药物和含胺脂质。在一个实施方案中,含胺配体R4为含胺脂质,该脂质包括单烃尾或双烃尾。在一个优选的实施方案中,该脂质为具有双烃尾的磷脂。
在另一实施方案中,该亲水性聚合物R1可选自聚乙烯吡咯烷酮、聚乙烯基甲基醚、聚甲基噁唑啉、聚乙基噁唑啉、聚羟基丙基噁唑啉、聚羟基丙基-甲基丙烯酰胺、聚甲基丙烯酰胺、聚二甲基-丙烯酰胺、聚羟基丙基甲基丙烯酸酯、聚羟基乙基丙烯酸酯、羟甲基纤维素、羟乙基纤维素、聚乙二醇、聚天冬酰胺、它们的共聚物,以及聚环氧乙烷-聚环氧丙烷。
在一个优选的实施方案中,该亲水性聚合物R1为聚乙二醇。
在另一个实施方案中,当R1为聚乙二醇时,R5为H而R2为CH3或C2H5。
在另一实施方案中,该含胺配体R4为多肽。在另一实施方案中,该多肽可以是重组多肽。示范性的和优选的多肽包括细胞因子如干扰素、白介素、生长因子和酶。
其中R1为含有用于连接到二硫代苄基部分的键的亲水性聚合物;R2选自H、烷基和芳基;R3选自O(C=O)R4、S(C=O)R4和O(C=S)R4;R4包括离去基团;而R5选自H、烷基和芳基;其中CH2-R3的取向选自邻位和对位。该组合物还包括药学上可接受的载体如盐水、缓冲剂等等。
在这方面的另一实施方案中,R2选自CH3,C2H5和C3H8。
在另一实施方案中,R3为O(C=O)R4而R4为含羟基或含氧的离去基团。在另一实施方案中,该离去基团源于选自氯化物、对硝基苯酚、邻硝基苯酚、N-羟基-四氢邻苯二甲酰亚胺、N-羟基琥珀酰亚胺、N-羟基戊二酰亚胺、N-羟基降冰片烯-2,3-二羧基亚胺、1-羟基苯并三唑、3-羟基吡啶、4-羟基吡啶、2-羟基吡啶、1-羟基-6-三氟甲基苯并三唑、咪唑、三唑、N-甲基-咪唑、五氟苯酚、三氟苯酚和三氯苯酚的化合物。
在另一实施方案中,所要求的化合物与代替R4的含胺配体反应形成包括该含胺配体的缀合物。例如该含胺配体可以是磷脂。
在一个优选的实施方案中,该亲水性聚合物R1为聚乙二醇,R5为H而R2为CH3或C2H5。
在该实施方案的另一方面,含有该缀合物的组合物包括脂质体。该脂质体可以进一步包括截留治疗剂。
在另一实施方案中,该含胺配体包括多肽。
其中R1为含有用于连接到二硫代苄基部分的键的亲水性聚合物;R2选自H、烷基和芳基;R3选自O(C=O)R4、S(C=O)R4和O(C=S)R4;R4包括含胺配体;而R5选自H、烷基和芳基;其中CH2-R3的取向选自邻位和对位。该脂质体与具有通过脂族二硫键连接到脂质体上的亲水性聚合物链的脂质体相比,具有更长的血液循环寿命。
在另一实施方案中,该脂质体进一步包括截留治疗剂。
在另一方面,本发明包括一种改善具有可释放亲水性聚合物链的表面包被脂质体的血液循环寿命的方法。该方法包括制备具有约1%-约20%的具有以下通式结构的化合物的脂质体:
其中R1、R2、R3和R5如以上所述,R4包括含胺脂质。
在这方面的一个优选的实施方案中,R5为H而R2选自CH3、C2H5和C3H8。
在另一实施方案中,该含胺脂质包括磷脂而R1为聚乙二醇。
在这方面,该脂质体可以进一步包括截留治疗剂。
结合附图阅读以下本发明的详细描述将更为全面地理解本发明的这些和其它目的及特征。
附图的简要说明
图1A显示了本发明的一个实施方案,其中二硫代苄基(DTB)连接甲氧基-聚乙二醇(mPEG)部分和含胺配体;
图1B显示了图1A的化合物硫解断裂后的产物;
图2说明了合成mPEG-DTB-胺-脂质的合成反应方案,其中该胺-配体为脂质二硬脂酰磷脂酰乙醇胺(DSPE);
图3说明了对二硫代苄基尿烷(DTB)-连接的mPEG-DSPE缀合物的硫解断裂机理;
图4A-4B显示了根据本发明制备mPEG-DTB-DSPE化合物的合成反应方案,其中DTB键被烷基位阻;
图5显示了根据本发明制备mPEG-DTB-配体化合物的另一合成反应方案;
图6A为合成mPEG-DTB-脂质的合成反应方案,它通过硫解断裂产生阳离子脂质;
图6B显示了图6A的化合物硫解断裂后的产物;
图7A显示了邻mPEG-DTB-DSPE和对mPEG-DTB-DSPE缀合物在溶液中断裂以单独在缓冲剂中(邻缀合物(*)、对缀合物(+))和在150μM的半胱氨酸的存在下(邻缀合物(空心圆)、对缀合物(空心正方形))产生胶束的速率;
图7B显示了图7A所述的胶束mPEG-DTB-DSPE缀合物和在脂质体中形成并在150μM的半胱氨酸的存在下温育的邻mPEG-DTB-DSPE(实心圆)和对mPEG-DTB-DSPE(实心正方形)缀合物的断裂速率;
图8A-8B显示了在指示浓度的半胱氨酸的存在下温育的含有DOPE:邻mPEG-DTB-DSPE(图8A)或DOPE:对mPEG-DTB-DSPE(图8B)的脂质体上释放的截留荧光团的含量百分率;
图9A显示了含有DOPE和对mPEG-DTB-DSPE的脂质体的归一化的截留荧光团释放百分率对时间的函数。截留荧光团的释放百分率是相对于不存在半胱氨酸时温育的脂质体上荧光团的释放百分率而归一化的。显示了在存在15μM(实心正方形)、75μM(空心三角形)、150μM(X符号)、300μM(空心圆),1500μM(实心圆),3000μM(+符号)和15000μM(空心菱形)浓度的半胱氨酸的情况下温育的脂质体的释放率;
图9B显示了含有DOPE和对mPEG-MeDTB-DSPE的脂质体的归一化的截留荧光团释放百分率对时间的函数。截留荧光团的释放百分率是相对于不存在半胱氨酸时温育的脂质体上荧光团的释放百分率而归一化的。显示了在存在15μM(实心正方形)、75μM(空心三角形)、150μM(X符号)、300μM(空心圆),1500μM(实心圆),3000μM(+符号)和15000μM(空心菱形)浓度的半胱氨酸的情况下温育的脂质体的释放率;
图9C显示了含有DOPE和图6A的mPEG-meDTB-二硬脂酰-甘油化合物的脂质体的归一化的截留荧光团释放百分率对时间的函数。截留荧光团的释放百分率是相对于在不存在半胱氨酸时温育的脂质体上荧光团的释放百分率而归一化的。显示了在存在15μM(实心正方形)、75μM(空心三角形)、150μM(X符号)、300μM(空心圆),1500μM(实心圆),3000μM(+符号)和15000μM(空心菱形)浓度的半胱氨酸的情况下温育的脂质体上断裂出化合物时染料的释放率;
图10显示了在注射含有PHPC∶胆固醇∶mPEG-DTB-DSPE(55∶40∶5摩尔比率)的脂质体的不同时间后的小鼠身上所取血样中脂质体的数量,以每分钟计数/ml含有截留In111的脂质体表示。一组动物在时间零(实心正方形)接受200μL注射的200mM半胱氨酸。对照组在时间零(空心圆)时注射盐水;
图11A显示了根据本发明的另一实施方案合成mPEG-DTB-蛋白质化合物的合成反应方案;
图11B显示了图11A的化合物硫解断裂后的分解产物;
图12为与mPEG-MeDTB-氯甲酸硝基苯基酯反应15分钟(泳道1)或1小时(泳道2)形成mPEG-MeDTB-溶菌酶缀合物的溶菌酶、天然溶菌酶(泳道3)、与mPEG-氯甲酸硝基苯基酯反应1小时的溶菌酶(泳道4)、分子量标志(泳道5)和用2%β-巯基乙醇在70℃下处理10分钟的泳道1-4的试样(泳道6-9)的十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)曲线的示意图;
图13显示了mPEG-DTB-p-硝基酰苯胺缀合物硫解断裂后的分解产物;
图14A显示了mPEG-MeDTB-对硝基酰苯胺(封闭的菱形)和在体外用5mM半胱氨酸温育2分钟(封闭的正方形)、5分钟(x符号)、10分钟(空心正方形)、20分钟(三角形)、40分钟(空心菱形)和80分钟(封闭的圆)后的吸光度对波长的函数,单位nm;
图14B显示了在体外在5mM半胱氨酸(封闭的圆)、1mM半胱氨酸(封闭的正方形)和0.15mM半胱氨酸(封闭的菱形)的存在下温育的mPEG-MeDTB-对硝基酰苯胺缀合物上释放的对硝基酰苯胺数量(单位:mole/L)对时间(单位:分钟)的函数。
发明详述
I.定义
这里所用的“多肽”指氨基酸聚合物,而不指氨基酸聚合物的具体长度。因此,如术语肽、寡肽、蛋白质和酶包括在多肽的定义范围内。该术语还包括多肽的表达后修饰如糖基化、乙酰化、磷酸化等等。
“含胺”意指任何这样的化合物,它们具有通过由烷基或芳基取代一或两个氢原子得到通式结构RNH2(伯胺)和R2NH(仲胺)而得自氨的部分,其中R为任何烃基。
这里的“亲水性聚合物”指具有水溶性部分的聚合物,该水溶性部分赋予该聚合物在室温下一定程度的水溶性。亲水性聚合物的例子包括聚乙烯吡咯烷酮,聚乙烯基甲基醚,聚甲基噁唑啉,聚乙基噁唑啉,聚羟基丙基噁唑啉,聚羟基丙基-甲基丙烯酰胺,聚甲基丙烯酰胺,聚二甲基-丙烯酰胺,聚羟基丙基甲基丙烯酸酯,聚羟基乙基丙烯酸酯,羟甲基纤维素,羟乙基纤维素、聚乙二醇、聚天冬酰胺、上述聚合物的共聚物以及聚环氧乙烷-聚环氧丙烷共聚物。在美国专利号5,395,619和5,631,018中描述了这些聚合物中的许多的性质和反应。
“包括反应官能团的聚合物”或“包括用于连接的键的聚合物”指已在用于与另一种化合物反应形成共价键的末端部分被修饰(通常但不是必需的)的聚合物。官能化聚合物以具有这样的一个反应官能团部分的反应方案可由本领域技术人员容易地确定和/或已在例如美国专利号5,613,018或由Zalipsky等人在例如Eur.Polymer.J.,19(12):1177-1183(1983);Bioconj.Chem.,4(4):296-299(1993)所描述。
“重组多肽”中的“重组”意指通过实验操作将氨基酸结合到期望的序列上。
这里所用的“烷基”意指来源于烷烃,通过从任何碳原子除去氢原子得到的基团:“CnH2n+1”。由不分枝的烷烃的末端碳原子上除去氢原子而得到的基团形成了正常烷基(正烷基)的亚组H[CH2]n。基团RCH2-、R2CH-(R不为H)和R3C-(R不为H)分别为伯、仲和叔烷基。
“芳基”指具有单环(如苯)或两个稠合环(如萘基)的取代或未取代的一价芳族基。该术语包括杂芳基,它们是在环上具有一或多个氮、氧或硫原子的芳环基,如呋喃基、吡咯、吡啶基和吲哚。“取代”指该芳基上的一或多个环氢被卤化物如氟、氯或溴取代;被含一或两个碳原子的低级烷基取代;被硝基、氨基、甲氨基、二甲氨基、甲氧基、卤甲氧基、卤甲基或卤乙基取代。
“脂族二硫化物”键指R′-S-S-R″形式的键,其中R′和R″为可被进一步取代的直链或支链烷基链。
这里采用以下的缩写:PEG,聚(乙二醇);mPEG,甲氧基-PEG;DTB,二硫代苄基;MeDTB,甲基-二硫代苄基;EtDTB,乙基-二硫代苄基;DSPE,二硬脂酰磷脂酰乙醇胺;DOPE,二油酰磷脂酰乙醇胺;PHPC,部分氢化的磷脂酰胆碱;MALDI-TOFMS,基体辅助激光解吸/电离时间飞行质谱。
II.本发明化合物
其中R1包括一种亲水性聚合物,该亲水性聚合物含有适宜将该聚合物共价结合到二硫代苄基部分的官能团。R2和R5独立地选自H、烷基或芳基,并可见它可变化以配合二硫化物的断裂速率。例如,为了获得更快的断裂速率,R2和R5为氢。通过为R2和R5之一或两者选择烷基或芳基对二硫化物进行位阻而获得较慢的断裂速率。R3包括与R4连接的连接部分,它包括含胺配体。在优选的实施方案中,该连接部分为O(C=O)、S(C=O)或O(C=O)。该含胺配体R4可以是伯或仲胺并可以选自任何数量的底物,包括但不限于脂质、药物、多肽、病毒、生物材料的表面和氨基糖苷。在优选的实施方案中,R4为含伯或仲胺的脂质、药物或多肽。在本发明的化合物中,基团CH2-R3的取向可以是邻或对位。
图1A显示了本发明的示范化合物的结构,其中R1为亲水性聚合物甲氧基-聚乙二醇、mPEG=CH3O(CH2CH2O)n,其中n为约10-约2300,其对应的分子量为约440道尔顿-约100,000道尔顿。该聚合物的分子量在一定程度上取决于R3的选择。在R3为用于脂质体的含胺脂质的实施方案中,优选的PEG分子量范围为约750-约10,000道尔顿,更优选为约2,000-约5,000道尔顿。该实施方案中的mPEG包括尿烷连接部分。在R3为含胺多肽的实施方案中,优选的PEG分子量范围为约2,000-约40,000道尔顿,更优选为约2,000-约20,000道尔顿。可以理解,R1可选自各种亲水性聚合物,而示范性的聚合物如上述。还可以理解,对某些配体如多肽,聚合物的分子量可取决于结合到该配体上的聚合物链的数量,其中在结合的聚合物链数量较小时通常选择更大分子量的聚合物。
继续参照图1a,在该示范性化合物中的R2和R5为H,但是R2和R5之一或二者还可以是直链或支链烷基或芳基。在一个优选的实施方案中,R5为H而R2为烷基,以下给出一些实施例。在图1A所示的化合物中,R3取通式O(C=O)-(NH2-配体),其中该NH2-配体可以是任何含胺多肽、药物或脂质,以下给出各实施方案的特定实施例。R3还可以是式O(C=S)-(NH2-配体)或S(C=O)-(NH2-配体)。
图1B显示了图1A的mPEG-DTB-(NH2-配体)化合物的硫解断裂的机理。氨基甲酸邻或对二硫代苄基酯部分在温和的硫解条件下如在半胱氨酸或其它天然还原剂的存在下可断裂。在断裂时,含胺配体再生其天然的未修饰的形式。以下所述的支持本发明的研究表明天然的体内生理学条件足以引发和实现DTB键的断裂。考虑该还原剂还可给药至足以断裂和分解该化合物的人工诱发硫解条件。
如上述,R3的一般形式为连接部分,如结合到含胺配体上的O(C=O)、S(C=O)或O(C=S)。在优选的实施方案中,该含胺配体包括含胺多肽、药物或脂质。现在将描述这些实施方案的实施例
A.含胺脂质
在一个实施方案中,该含胺配体为含胺脂质。这里所指的脂质意指具有至少一个含有至少约8个碳原子的酰基链,更优选含约8-24个碳原子的酰基链的不溶于水的分子。优选的脂质为具有含胺极性头基团和酰基链的脂质。示范性的脂质为具有单酰基链(如硬脂酰胺)或两个酰基链的磷脂。优选的具有含胺头基团的磷脂包括磷脂酰乙醇胺和磷脂酰丝氨酸。该脂质尾部可以具有约12-约24个碳原子,并可以是完全饱和或不饱和的。一种优选的脂质为二硬脂酰磷脂酰乙醇胺(DSPE),但是本领域技术人员将考虑落在该说明书范围内的大量脂质。还考虑该脂质可天然地包含胺基或可以衍生为包括胺基。其它不具酰基尾部的脂质部分如胆固醇胺也是合适的。
聚合物-DTB-脂质化合物的合成如图2的图示。通过2-(甲氧基羰基二硫代)乙烷胺与mPEG-氯甲酸酯的反应而制得具有甲氧基羰基二硫代烷基末端基团的mPEG衍生物(MW 2000和5000道尔顿),该mPEG-氯甲酸酯已通过光气化干燥的mPEG-OH溶液(Zalipsky,S.,等人,Biotechnol.Appl.Biochem.15:100-114(1992).)制得。根据公开的方法(Brois,S.J.,等人,J.Amer.Chem.Soc.92:7629-7631(1970);Koneko,T.,等人,Bioconjugate Chem.2:133-141(1991)),通过2-氨基乙硫醇盐酸盐与等量的甲氧基羰基硫化氯的反应得到前一化合物。巯基苄基醇的对和邻位异构体(Grice,R.,等人,J.Chem.Soc.1947-1954(1963))都干净地与所得的PEG-连接的酰基二硫化物偶合,得到含有二硫代苄基醇末端基团的mPEG。活性碳酸盐的引入与采用未衍生的mPEG-OH一样进行,得到对硝基苯基碳酸酯。在乙醇胺中加入DSPE形成期望的mPEG-DTB-DSPE产物。通过硅胶色谱制备和纯化邻和对DTB-脂质化合物,用NMR和MALDI-FOFMS进行表征,其具体情况由实施例1给出。
图3显示了mPEG-DTB-DSPE缀合物的硫解断裂机理。在断裂时,磷脂酰乙醇胺脂质再生其天然、未修饰的形式。
图4A-4B显示了用于合成具有与二硫键(例如更受阻的二硫键)相邻的烷基的mPEG-DTB-DSPE缀合物的反应方案。如在实施例2A中更为全面的描述,二氯甲烷中的mPEG-OH在三乙胺(TEA)的存在下与氯甲酸对硝基苯基酯反应形成mPEG-碳酸硝基苯基酯。氨基醇如在二甲基甲酰胺(DMF)中的1-氨基-2-丙醇或1-氨基-2-丁醇在TEA的存在下与mPEG-碳酸硝基苯基酯反应形成连接到PEG上的仲醇。然后将该仲醇转化为期望的如图4A所示的以在实施例2A中所述的mPEG-DTB-DSPE化合物。
在该反应方案中,mPEG-甲基-二硫代苯基-氯甲酸硝基苯基酯与DSPE反应形成期望的化合物。在该mPEG-甲基-二硫代苯基-氯甲酸硝基苯基酯化合物中的氯甲酸硝基苯基酯部分作为离去基团在与选择的脂质反应时产生期望的产物。本发明在另一方面构想了一种组合物,它包括通过与化合物如mPEG-甲基-二硫苄基-R3的反应而产生的化合物其中R3代表通过连接部分连接到苯环上的离去基团。该离去基团在与含胺配体如DSPE、多肽或含胺药物反应时被取代。该离去基团的选择根据配体中胺的反应性,并优选来自各种具有含羟基或含氧的离去基团的酸性醇。它们包括氯化物、对硝基苯酚、邻硝基苯酚、N-羟基-四氢邻苯二甲酰亚胺、N-羟基琥珀酰亚胺、N-羟基-戊二酰亚胺、N-羟基降冰片烯-2,3-二羧基亚胺、1-羟基苯并三唑、3-羟基吡啶、4-羟基吡啶、2-羟基吡啶、1-羟基-6-三氟甲基苯并三唑、咪唑、三唑、N-甲基-咪唑、五氟苯酚、三氟苯酚和三氯苯酚。
实施例2B描述了mPEG-EtDTB-脂质缀合物的制备,其中的二硫键被乙基部分位阻。
图5显示了根据本发明制备mPEG-DTB-配体化合物的另一合成反应方案。该反应过程的细节在实施例3A-3B中给出。简言之,冷1-氨基-2-丙醇与硫酸反应形成硫酸2-氨基-1-甲基乙基氢酯。该产物在含水乙醇中与二硫化碳和氢氧化钠反应形成5-甲基噻唑烷-2-硫酮。往该5-甲基噻唑烷-2-硫酮中加入盐酸水溶液并加热。在回流一周后,结晶并回收产物1-巯基(甲基)乙基氯化铵。产物与甲氧基羰基硫化氯反应得到2-(甲氧基羰基二硫代)乙烷胺。采用上述图2的方法进行的2-(甲氧基羰基二硫代)乙烷胺与mPEG-氯甲酸酯的反应获得期望mPEG-DTB-硝基苯基化合物,该化合物适于与选择的含胺配体反应形成本发明的化合物。
实施例3B描述了合成mPEG-(乙基)DTB硝基苯基的反应。
图6A显示了根据本发明的制备另一种mPEG-DTB-脂质化合物的反应方案。该反应的细节由实施例4提供。将脂质1,2-二硬脂酰-sn-甘油活化以与按图4A或图5所述制备的mPEG-DTB-硝基苯基反应。所得的mPEG-DTB-脂质与上述化合物不同之处在于缺乏磷酸酯头基团。图6A的mPEG-DTB-脂质在断裂前为中性。如在图6B中所示,在硫解还原该二硫键时,该化合物分解形成阳离子脂质。带正电的脂质在体内提供静电相互作用并同样有助于在体内的靶向。
在上述的反应方案中,所要求的化合物的R5为H。但是,在另一实施方案中,R5为烷基或芳基部分。例如,在该方法中,R2和R5都为CH3部分,α,β-未饱和的酰基氯(R’R”C=CHCOCl,其中R’为例如CH3而R”为CH3,但可考虑任何烷基或芳基)与胺结尾的PEG反应得到对应的N-PEG-取代的α,β-未饱和的酰胺。该化合物与硫羟乙酸反应,通过将缀合物加到C=C键上而得到对应的N-PEG-取代的β-(乙酰硫基)酰胺。将该乙酰硫基(-SCOCH3)水解成硫羟基(-SH),然后与甲基(氯亚磺酰)甲酸酯(CISCOOCH3)反应,产生甲氧基羰基二硫基(-SSCOOCH3);然后将该中间产物与对巯基苄基醇反应得到N-PEG-取代的β-(二硫代苄基醇)酰胺(具有结构PEG-NH-CO-CH2CR’R”-SS-p-苯基-CH2OH)。然后将该苄基醇部分与氯甲酸硝基苯基酯反应得到上述的碳酸硝基苯基酯离去基团。
1.mPEG-DTB-DSPE化合物的体外断裂
通过在缓冲的水溶液(pH7.2)中制备化合物的胶束溶液来研究邻mPEG-DTB-DSPE和对mPEG-DTB-DSPE(按实施例1所述制备)的体外断裂。如实施例5所述,在存在和不存在150μM半胱氨酸下通过HPLC分析该化合物的消失来监测该化合物的硫解断裂。结果如在图7A中所示,其中邻和对位化合物在不存在半胱氨酸(分别为*符号和+符号)下没有显示出断裂并在这些条件下在不存在半胱氨酸时表现稳定。图7A中显示了邻和对位化合物在存在150μM半胱氨酸下的断裂,分别由空心圆和空心正方形表示。该邻位化合物表现出比对位对应物稍快的分解速率(T1/2≈12分钟对≈18分钟)。
2.含有mPEG-DTB-脂质化合物的脂质体组合物
a.体外表征
在一个实施方案中,将mPEG-DTB-脂质化合物制成脂质体。脂质体为封闭的脂质囊,用于各种治疗目的,特别是通过脂质体的系统给药而携带治疗剂至靶区域或细胞。具体而言。具有亲水性聚合物链如聚乙二醇(PEG)的表面包被的脂质体为期望的药物载体,因为这些脂质体与缺少该聚合物包被的脂质体相比延长了血液循环寿命。该聚合物包被中的聚合物链屏蔽了脂质体并形成该脂质体的水溶剂化聚合物链周围的“刚性刷”。因此,该聚合物用作血液蛋白质的屏障,防止蛋白质的结合和脂质体的识别而被巨噬细胞和其它网状内皮系统细胞摄取和除去。
具有聚合物链的表面包被的脂质体一般通过在脂质混合物中包含约1-约20摩尔百分率的由该聚合物衍生的脂质而制得。聚合物衍生的脂质的实际数量可以更高或更低,取决于该聚合物的分子量。在本发明中,通过往其它脂质体脂质双层组分中加入约1-约20摩尔百分率的聚合物-DTB-脂质缀合物而制备脂质体。如在以下的研究中所示,本发明的含有聚合物-DTB-脂质缀合物的脂质体比含有聚合物-脂质缀合物,且其中的聚合物和脂质通过可断裂的脂肪族二硫键结合的脂质体具有更长的血液循环寿命。
在支持本发明的研究中,如在实施例6中所述制备包括成囊脂质部分氢化的磷脂酰胆碱和胆固醇以及邻mPEG-DTB-DSPE或对mPEG-DTB-DSPE化合物的脂质体。在存在和不存在处于含水缓冲剂的150μM半胱氨酸下监测半胱氨酸介导的mPEG-DTB-DSPE化合物的断裂。结果如图7B所示,它包括图7A的数据以进行比较。在图7B中,不存在半胱氨酸(分别为*符号和+符号)下,胶束形式邻和对位化合物没有显示断裂,它表明该缀合物在不存在硫醇时的稳定性。空心圆和空心正方形分别对应于上面图7A所讨论的在存在半胱氨酸下胶束形式的邻和对位化合物。实心圆和实心正方形分别对应于在半胱氨酸的存在下的脂质体形式的邻和对位化合物。
图7B中的数据表明邻和对位化合物在被掺到脂质体中时稍微更耐受硫解断裂。硫解反应产物的TLC(硅胶G,氯仿/甲醇/水90∶18∶2)(Dittmer,J.C.,等人,J.lipid Res.5:126-127(1964))检查表明DSPE是唯一的脂质组分而另一点对应于含硫醇、不含脂质的mPEG衍生物。
在支持本发明的另一研究中,由脂质二油酰磷脂酰乙醇胺(DOPE)制得脂质体并制得邻mPEG-DTB-DSPE或对mPEG-DTB-DSPE化合物。DOPE为六角形相脂质,它单独不形成脂质囊。但是,当DOPE与少量摩尔百分率的mPEG-DTB-DSPE化合物结合时将形成脂质体。mPEG-DTB-DSPE化合物的断裂引发脂质体的分解和脂质体-截留含量的释放。因此,这种脂质体的含量释放特征为可断裂的含PEG的脂质体提供了一种方便的定量评价。
如实施例7A所述采用截留荧光团、对二甲苯-双-溴化吡啶鎓和8-羟基芘三磺酸三钠制得含有DOPE和邻或对mPEG-DTB-DSPE化合物的脂质体。如实施例7B所述监测在各种浓度的半胱氨酸的存在下温育的脂质体上的荧光团的释放。
含有邻位化合物的脂质体的结果如图8A所示,其中显示了在15μM(实心菱形)、150μM(实心反转的三角形)、300μM(实心三角形)和1.5mM(实心圆)浓度的半胱氨酸的存在下温育的脂质体上的截留荧光团的含量释放百分率。图8B为含有对位化合物的脂质体的类似的图形,其中在15μM(实心菱形)、300μM(实心三角形)、1μM(实心正方形)和1.5mM(实心圆)浓度的半胱氨酸中温育该脂质体。
图8A-8B显示了邻和对位化合物在被掺到脂质体内时都断裂,它可由速度取决于半胱氨酸浓度的截留染料的释放来证明。用含有不可断裂的mPEG-DSPE的脂质体的对照研究没有产生含量释放(这里没有显示结果)。这些结果还表明邻位缀合物某种程度上更易于硫解断裂,例如,300μM的半胱氨酸在20分钟内释放出绝大部分含量的DOPE脂质体。在同一条件下,只有部分具有对mPEG-DTB-DSPE的脂质体分解。类似地,在150μM的半胱氨酸下温育20分钟后,含有邻位化合物的脂质体释放出半数的截留含量,而含有对位化合物的脂质体只有约10%的含量释放。邻位和对位化合物的半衰期在半胱氨酸水平为150μM时都小于20分钟(见图7B中的数据)。这表明半数以上的最初三摩尔百分率的mPEG-DTB-脂质必须断裂以从脂质体上观测含量释放。
mPEG-DTB-DSPE/DOPE脂质体在15μM的半胱氨酸中的分解、人和啮齿动物身上的平均血浆浓度(Lash,L.H.,等人,Arch.Biochem.Biophys.240:583-592(1985))在这些实验的时间范围内(60分钟)最小。这表明mPEG-DTB-脂质化合物在血浆中应具有足够长的寿命以使PEG移植囊在体内的系统分布或在特定的部位被动地或通过配体介导的靶向蓄积。半胱氨酸浓度的局部或短期增大可由其静脉内的或动脉内给药有效地实现。图8A-8B所示的结果还表明延长与天然血浆半胱氨酸浓度(≈15μM)的接触将足以分解大部分的这些化合物。在体内实验中研究这些建议,并如下所示。
在另一支持本发明的研究中,制备含有DOPE和三种不同mPEG-DTB-脂质化合物的脂质体。该脂质体如实施例7所述制备并包括和截留荧光团。三种mPEG-DTB-脂质化合物为图1A中所示的mPEG-DTB-DSPE、图4B中所示的mPEG-MeDTB-DSPE,其中R为CH3,和图6A中所示的mPEG-MeDTB-二硬脂酰甘油。该脂质体含有97摩尔百分率的DOPE和3摩尔百分率的一种mPEG-DTB-脂质化合物。在各种半胱氨酸浓度下通过监测截留荧光团的释放作为时间的函数而测定由半胱氨酸介导的该化合物的断裂速率。结果如在图9A-9C中所示,其中截留荧光团的释放百分率相对于在单独在缓冲剂中温育的脂质体的释放速率进行归一化。
图9A显示了含有DOPE和对mPEG-DTB-DSPE(图1A的化合物)的脂质体的截留荧光团的释放百分率作为时间的函数。显示了含有该缀合物并在15μM(实心正方形)、75μM(空心三角形)、150μM(X符号)、300μM(空心圆)、1500μM(实心圆)、3000μM(+符号)和15000μM(空心菱形)浓度的半胱氨酸的存在下温育的脂质体上的释放率。
图9B显示了含有DOPE和对mPEG-MeDTB-DSPE(图4B的化合物)的脂质体的截留荧光团的释放百分率作为时间的函数。显示了在15μM(实心正方形)、75μM(空心三角形)、150μM(X符号)、300μM(空心圆)、1500μM(实心圆)、3000μM(+符号)和15000μM(空心菱形)浓度的半胱氨酸的存在下温育的脂质体上荧光团的释放率。
图9C为采用DOPE和mPEG-MeDTB-二硬脂酰甘油(图6A的化合物)制成的脂质体的类似的图形。显示了在15μM(实心正方形)、75μM(空心三角形)、150μM(X符号)、300μM(空心圆)、1500μM(实心圆)、3000μM(+符号)和15000μM(空心菱形)浓度的半胱氨酸的存在下温育的脂质体上染料的释放率。
图9A-9C表明mPEG-MeDTB-脂质断裂的速率取决于半胱氨酸浓度,由截留荧光的释放表明在15-75μM的半胱氨酸浓度下的断裂速度缓慢。对比图9A和图9B中的数据,表明mPEG-MeDTB-DSPE化合物(图9B)的断裂比mPEG-DTB-DSPE化合物(图9A)约慢10倍。因此,可以根据DTB键中的R部分(见图2)调节断裂的速率。
b.体内表征
在小鼠身上测定如在实施例8所述制备并包括本发明的聚合物-DTB-脂质缀合物的脂质体的血液循环寿命。将In111截留在脂质体内并通过静脉内注射给予该脂质体。一组受试动物额外接受注射半胱氨酸,对照组动物额外接受注射盐水。在各时间处取血样并分析脂质体的存在,这种存在由In111的存在来证明。
图10显示了结果,其中显示了每分钟In111的计数(CPM)作为作为注射脂质体和盐水(空心圆)或200mM半胱氨酸(实心正方形)后的时间的函数。可见,mPEG-DTB-DSPE在暴露于天然生理条件下发生断裂,这由在给予脂质体后用盐水处理的小鼠的组中的断裂来证明。对小鼠给予外源性还原剂、半胱氨酸有效地增大在约2-约8小时的时间范围内mPEG-DTB-脂质化合物的断裂速率。
重要的是,本发明聚合物-DTB-脂质化合物的断裂导致再生初始的未修饰形式的脂质。这是期望的,因为非天然的、修饰的脂质可能具有不期望的体内作用。同时该化合物在不存在还原剂的情况下储存时稳定。
在这里没有显示的其它研究中,对比了含有mPEG-DTB-脂质的脂质体和含有聚合物-脂质缀合物的脂质体的血液循环寿命,在含有聚合物-脂质缀合物的脂质体中,聚合物和脂质通过可断裂的脂肪族二硫键结合。脂肪族二硫键容易在体内断裂,而具有通过脂肪族二硫化物移植到其表面的聚合物链的脂质体的血液循环寿命一般不具有在含稳定连接的聚合物链的脂质体上观察到的延长的血液循环寿命。本发明的二硫醇苄基键,尤其是更受阻的DTB键在体内更稳定且与具有通过脂肪族二硫键结合的聚合物链的脂质体相比获得了更长的血液循环寿命。
B.含胺多肽
在另一实施方案中,本发明包括图1A所述的化合物,其中该含胺配体为多肽。图11A显示了表明制备聚合物-DTB-多肽的合成反应方案,其中mPEG作为示范性的聚合物。一般而言,根据上述图2、4A和5中所述的合成途径之一制备mPEG-DTB-离去基团化合物。该离去基团可以是碳酸硝基苯基酯或上述其它之任一种。该mPEG-DTB-碳酸硝基苯基酯化合物通过尿烷键偶联到多肽中的胺部分。与化合物中的二硫化物邻近的R基团可以是H、CH3、C2H5等等,并可根据所期望的二硫化物断裂速率来选择。
图11B显示了在化合物的半胱氨酸介导的断裂时的分解产物。可见天然蛋白质(没有修饰其蛋白质胺基)在断裂时发生再生。
聚合物链如PEG与多肽的连接经常减小多肽的酶或其它生物活性,例如受体结合。但是,多肽的聚合物修饰增加了多肽的血液循环寿命。在本发明中,给予受试者聚合物修饰的多肽。由于聚合物修饰的多肽循环,接触生理还原条件如血液半胱氨酸和其它体内硫醇引发了多肽上聚合物链的断裂。在聚合物链从多肽上释放时,逐渐地恢复多肽的生物学活性。通过这种方式,多肽最初具有足够的用于生物分布的血液循环寿命,并在聚合物链断裂时在一定期间内恢复其全部生物活性。
在一个支持本发明的研究中,溶菌酶用作模型多肽,并通过与上述类似的合成途径制备了mPEG-MeDTB-溶菌酶缀合物。在0.1M硼酸盐中,在pH9和2∶1的碳酸硝基苯基酯与溶菌酶的氨基的比率下温育该溶菌酶。在反应15分钟和3小时后,由SDS-PAGE表征试样。通过在相同的条件下溶菌酶与mPEG-碳酸硝基苯基酯缀合物反应60分钟制备对比化合物,它将形成稳定的mPEG-溶菌酶缀合物。
图12显示了SDS-PAGE凝胶的示意图。泳道1对应于在溶菌酶与mPEG-MeDTB-碳酸硝基苯基酯反应15分钟后形成的化合物,而泳道2对应于在同样的化合物反应1小时后形成的化合物。泳道3代表天然溶菌酶而泳道4对应于与mPEG-碳酸硝基苯基酯反应1小时的溶菌酶。从上向下泳道5中分子量标记如下:
分子量(k道尔顿) | 标记 |
1163 | β-半乳糖苷酶 |
97.4 | 磷酸化酶b |
66.3 | 牛血清白蛋白 |
55.4 | 谷氨酸脱氢酶 |
36.5 | 乳酸脱氢酶 |
31 | 碳酸酐酶 |
21.5 | 胰蛋白酶抑制剂 |
14.4 | 溶菌酶 |
比较泳道1和泳道2表明,更长的反应时间导致化合物分子量的增大,并与在更长的温育时间内缀合到多肽上的其它mPEG链一致。
SDS-PAGE曲线的泳道6-9对应于在70℃下用2%β-巯基乙醇处理10分钟后的泳道1-4中的试样。mPEG-MeDTB-溶菌酶缀合物在与还原剂接触后分解再生天然溶菌酶,这可由在14.4kDa处的泳道6和7中的带证明。与此相对照,在用还原剂温育时并不影响稳定的mPEG-溶菌酯化合物,这可由泳道9和泳道4曲线上的一致性证明。
从SDS-PAGE曲线还是清楚可见的是,mPEG-MeDTB与蛋白质的共价结合形成含有各种mPEG-蛋白质比率的缀合物的混合物。该比率取决于反应时间和条件。它可由观察泳道1和2上的带而清楚可见,其中泳道1表示由约1-6个PEG链衍生的溶菌酶。在泳道2中,更长的反应时间获得具有更高mPEG-蛋白质比率的mPEG-MeDTB-溶菌酶缀合物。所有可断裂的缀合物易于断裂再生天然蛋白质,这可由泳道6和7上的带表明。
考虑使用上述的任何亲水性聚合物。该聚合物分子量的选择取决于多肽、多肽上反应性胺的数量和期望的该聚合物修饰的化合物的大小。
考虑使用的多肽是非限制性的并可以是天然或重组产生的多肽。优选小的人重组多肽,并优选范围为10-30KDa的多肽。示范性的多肽包括细胞因子,如肿瘤坏死因子(TNF)、白介素和干扰素、红细胞生成素(EPO)、粒细胞集落刺激因子(GCSF)、酶等等。还考虑病毒多肽,其中将该病毒的表面修饰为包括一或多种通过DTB可逆键连接的聚合物链。修饰含有用于细胞转染的基因的病毒将延长病毒的循环时间并减小它的免疫原性,借此改善外源性基因的递送。
C.含胺药物
在本发明的另一实施方案中,考虑聚合物-DTB-含胺药物形式的化合物。该化合物的结构如上所述,尤其是根据图1A所述,其中该图中的含胺配体为含胺药物。用PEG修饰治疗药物有效地改善药物的血液循环寿命并减少任何免疫原性。
根据上述任一反应方案制备聚合物-DTB-含胺药物,其中为特定的药物提供必要的修饰。大量的治疗药物具有反应性胺部分,如丝裂霉素C、博莱霉素、阿霉素和环丙氟哌酸,且本发明无限制性地考虑这些药物中任一种。还可考虑本发明还用于含有醇或羧基部分的药物。在该药物含有适于反应的羟基或羧基部分的情况下,该聚合物-DTB部分可以通过尿烷、酯、醚、硫醚或硫酯键连接到药物上。在所有这些实施方案中,该聚合物-DTB-药物化合物在体内给药后硫解分解而再生天然的活性形式的含胺药物,在修饰后和给药前的化合物的治疗活性不是必需的。因此,在给药和从药物上断裂DTB-聚合物之后用DTB-聚合物修饰该药物导致治疗活性的减小或丧失的情况下,再次得到该药物的活性。
在支持本发明的研究中,如图13所示,药物硝基酰苯胺与mPEG-MeDTB-碳酸硝基苯基酯反应形成mPEG-MeDTB-对硝基酰苯胺化合物。该化合物在与还原剂接触时分解产生图中所示的产物,其中该药物对硝基酰苯胺再生未修饰的状态。
在体外在含有5mM半胱氨酸的缓冲剂中温育该mPEG-MeDTB-对硝基酰苯胺化合物,并在图14A中显示了在各时间处提取的试样的吸光度。在图中可见在以下时间点处测得的试样:时间零(封闭的菱形)、2分钟(封闭的正方形)、5分钟(x符号)、10分钟(空心正方形)、20分钟(三角形)、40分钟(空心菱形)和80分钟(封闭的圆)。UV光谱的变化作为在半胱氨酸中温育时间的函数是明显的,它显示了半胱氨酸介导的mPEG-MeDTB-对硝基酰苯胺化合物上对硝基酰苯胺的释放。
图14B显示了在5mM半胱氨酸(封闭的圆)、1mM半胱氨酸(封闭的正方形)和0.15mM半胱氨酸(封闭的菱形)的存在下温育的mPEG-MeDTB-对硝基酰苯胺缀合物上释放的对硝基酰苯胺的数量,单位mole/L。缀合物上的药物释放速率取决于存在的还原剂的浓度。
根据前述可以看出如何满足本发明的各种目的和特征。本发明的化合物包括通过苄基尿烷键的邻或对位二硫化物而可逆地结合到亲水性聚合物上的含胺配体。该键在接受温和硫解条件时断裂再生初始的未修饰形式的含胺配体。可以通过该键中二硫化物的立体位阻和/或通过体内的硫解条件的控制来控制断裂的速率。在二硫代苄基键断裂前提供具有增大的血液循环寿命、改善的稳定性和降低的免疫原性的化合物。
III.实施例
以下的实施例进一步例举了这里所述发明,并不意图以任何方式限定本发明的范围。
材料
所有的材料商购自合适的销售商,如Aldrich公司。
实施例1
合成mPEG-DTB-DSPE
将mPEG-MeDTB-碳酸硝基苯基酯(300mg,0.12mmol,1.29eq)溶于CHCl3(3ml)。往PEG-溶液中加入DSPE(70mg,0.093mol)和TEA(58.5μl,0.42mmol,4.5eq),并在50℃(油浴温度)下搅拌。15分钟后,TLC表明反应没有完全。然后每10分钟后加入两部分的TEA(10μl,和20μl)和若干部分mPEG-MeDTB-碳酸硝基苯基酯(50mg,30mg,10mg),直至反应完全。蒸发溶剂。将反应混合物溶于MeOH,并加入1g C8二氧化硅。再次蒸发溶剂。将含有C8二氧化硅的产物加到柱顶部,并用以下溶剂洗脱:MeOH∶H2O梯度(压力)、MeOH∶H2O=30∶70,60ml;MeOH∶H2O=50∶50,60ml;MeOH∶H2O=70∶30,140ml(起始物质洗脱);MeOH∶H2O=75∶25=40ml;MeOH∶H2O=80∶20,80ml(洗脱产物);MeOH∶H2O=85∶15,40ml;MeOH∶H2O=90∶10,40ml;MeOH=40ml;CHCl3∶MeOH∶H2O=90∶18∶10,40ml。合并含纯产物的部分并蒸发得到无色浓液体产物。向其加入叔丁醇(5ml),冻干并在P2O5上真空干燥得到产物,为白色蓬松固体(252mg,89%产率)。
邻和对DTB-DSPE化合物通过硅胶色谱法(甲醇梯度0-10%在氯仿中,≈70%分离产率)纯化,并由NMR和MALDI-TOFMS确定其结构。(对位缀合物的1H NMR:(d6-DMSO,360MHz)δ0.86(t,CH3,6H),1.22(s,脂质的CH2,56H),1.57(m,CH2CH2CO2,4H),2.50(2xt,CH2CO2,4H),2.82(t,CH2S,2H),3.32(s,OCH3,3H),3.51(m,PEG,≈180H),4.07(t,PEG-CH2OCONH,2H),4.11 & 4.28(2 x dd甘油的CH2CH,2H),4.98(s,苄基-CH2,2H),5.09(m,脂质的CHCH2),7.35 & 7.53(2 x d,芳基,4H)ppm。该邻位缀合物的不同仅在于苄基和芳基信号在5.11(s,CH2,2H),和7.31(d,1H),7.39(m,2H)7.75(d,1H)ppm处。
MALDI-TOFMS产生以相同的44Da间隔分隔的离子的分布,对应于环氧乙烷重复单元。化合物的平均分子量对于对位和邻位异构体而言分别为3127和3139Da(理论分子量≈3100Da)。
反应方案如图2中所示。
实施例2
合成mPEG-DTB-DSPE
A.mPEG-MeDTB-DSPE
该反应方案如图4A-4B所示。用甲苯(总体积为270ml,250ml通过Dean-Stark法蒸馏出)共沸干燥mPEG(5K)-OH(40g,8mmol)。将二氯甲烷(100ml)加到mPEG-OH中。在4℃下(冰水)将氯甲酸对硝基苯基酯(2.42g,12mmol,1.5eq)和TEA(3.3ml,24mmol,3eq)加到PEG溶液,同时小心防止湿气。形成浅黄色TEA盐酸盐。15分钟后移去冷却浴,在室温下将反应混合物搅拌过夜。TLC表明(CHCl3∶MeOH∶H2O=90∶18∶2)反应完全。蒸发溶剂。将残留物溶于乙酸乙酯(50℃)。滤去TEA盐酸盐并用温热乙酸乙酯洗涤。蒸发溶剂并用异丙醇重结晶产物(三次)。产量:38.2g(92%)。1H NMR(DMSO-d6,360MHz)δ3.55(s,PEG,450H);4.37(t,PEG-CH2,2H);7.55(d,C6H5,2H);8.31(d,C6H5,2H)。
将1-氨基-2-丙醇(1.1ml,14.52mmol,3eq)和TEA(2.02ml,14.52mmol,3eq)加到在DMF(60ml)和CH2Cl2(40ml)中的mPEG(5K)-碳酸硝基苯基酯(25g,4.84mmol)。它是一种黄色清澈溶液。室温下搅拌反应混合物30分钟。TLC(CHCl3∶MeOH=90∶10)表明反应完全。蒸发溶剂(二氯甲烷)。往DMF(60ml)中的产物混合物中加入异丙醇(250ml)。产物立即沉淀,然后用iPrOH重结晶(三次)。产量:22.12g(90%)。1H NMR(DMSO-d6,360MHz)δ.98(d,CH3CH(OH)CH2,3H);3.50(s,PEG,180H);4.03(t,PEG-CH2,2H);4.50(d,CH3CHOH,1H);7.0(t,mPEG-OCONH)。
用甲苯(45ml)共沸干燥mPEG(5K)-尿烷-2-甲基丙醇(22.12g,4.34mmol)。往其加入二氯甲烷(60ml)。0℃下将甲烷磺酰氯(604.6μl,7.81mmol,1.8eq)和TEA(3.93ml,28.21mmol,6.5eq)加到mPEG-溶液,同时保持搅拌并小心防湿气。30分钟后移去冷却浴并在室温下搅拌反应混合物16小时。蒸发溶剂。加入乙酸乙酯除去TEA盐。产物用异丙醇重结晶(三次)。产量:20.27g(90%)。1HNMR(DMSO-d6,360MHz)δ1.27(d,CH3CHOSO2CH3,3H);3.162(s,CH3O2SOCH,3H);3.50(s,PEG,180H);4.07(t,PEG-CH2,2H);4.64(q,CH3CHOH,1H);7.43(t,mPEG-OCONH)。
将mPEG(5K)-尿烷-2甲基-甲烷砜(10.27g,1.98mmol)与甲苯(每次20ml)共沸干燥。0℃下将氢化钠(377mg,9.4mmol,4.75eq)加到无水甲苯(60ml)中(在冰水中)。5分钟后,三苯基甲烷硫醇(3.92g,14.6mmol,7.15eq)加到该溶液。10分钟后,将mPEG-尿烷-2-甲基-甲烷砜(10.27gm,1.98mmol)加到反应混合物。它成为黄色溶液。45分钟后,TLC(CHCl3∶MeOH∶H2O=90∶18∶2)表明反应完全。往该反应混合物中加入乙酸(445.57μl,7.42mmol,3.75eq)以中和过量的氢化钠。该溶液变稠和发白。蒸发溶剂并用乙酸乙酯(30ml)和异丙醇(70ml)重结晶固体。反应混合物没有完全溶解而被滤出。然后用异丙醇/叔丁醇(100ml/20ml)重结晶产物混合物。产量:8.87g(84%)。1H NMR(DMSO-d6,360MHz)δ.74(d,CH3CHSC(C6H5)3,3H),3.50(s,PEG,180H),4.0(t,PEG-CH2,2H),4.64(q,CH3CHOH,1H);7.49(t,mPEG-OCONH);7.20-7.41(m,SC(C6H5)3,15H)。
0℃下将mPEG(5K)-尿烷-2甲基-三苯基甲烷硫醇(8.87g,1.65mmol)溶于TFA/CH2Cl2(10ml/10ml)。剧烈搅拌下将甲氧基羰基硫化氯(185.5μl,1.99mmol,1.2eq)加到该溶液。室温下搅拌反应混合物15分钟。TLC(CHCl3∶MeOH=90∶10)表明反应完全。蒸发溶剂。用异丙醇∶叔丁醇(80ml∶20ml)重结晶产物混合物两次。往该产物中加入叔丁醇(5ml),然后冻干并在P2O5上真空干燥得到产物,为白色蓬松固体(8.32g,97%产率)。1H NMR(DMSO-d6,360MHz)δ1.17(d,CH3CHSSCOOCH3,3H);3.42(s,PEG,180H);3.84(s,CH3OCOSSCH,3H);4.05(t,mPEG-CH2,2H);7.38(t,mPEG-OCONH,1H)。
将mPEG(5K)-尿烷乙基(甲基)二硫代羰基甲醇盐(8.32g,1.6mmol)溶于干甲醇(20ml)和氯仿(2.5ml)。往该PEG-溶液中加入在干甲醇(2ml)中的巯基苄基醇溶液(592mg,4mmol,2.5eq)。室温下搅拌反应混合物18小时。蒸发溶剂,用乙酸乙酯/异丙醇,30ml/100ml重结晶产物混合物(3次)。NMR表明形成16%的产物。因此在0℃(冰水)下将另一部分在MeOH(2ml)中的巯基苄基醇(322mg,2.18mmol,1.8eq)滴加到在MeOH/CHCl3(24ml/1ml)中的产物混合物。完成加入(10分钟)后,除去冰浴,并在室温下搅拌反应混合物24小时。TLC(CHCl3∶MeOH∶H2O=90∶18∶2)表明反应完全。蒸发溶剂,然后用乙酸乙酯/异丙醇,30ml/100ml重结晶产物混合物。产量:7.25g,(94%)。1H NMR(DMSO-d6,360MHz)δ1.56(d,CH3CHSSC6H5CH2OH,3H);3.29(CH3O-PEG,3H);3.50(s,PEG,450H);4.03(t,mPEG-CH2,2H);4.46(d,HOCH2C6H5,2H);5.16(t,HOCH2C6H5,1H);7.30(d,C6H5,2H);7.40(br t,mPEG-OCONH,1H);7.50(d,C6H5,2H)。
将mPEG(5K)-尿烷-乙基(甲基)-二硫代苄基醇(6.75g,1.27mmol)溶于CHCl3(30ml)。0℃(冰水)往其加入氯甲酸对硝基苯基酯(513mg,2.54mmol,2eq)。5分钟后加入三乙胺(531μl,3.81mmol,3eq)。30分钟后除去冰浴,室温下搅拌反应混合物过夜。蒸发溶剂。将产物混合物溶于乙酸乙酯。滤出TEA盐,然后蒸发溶剂。随着用乙酸乙酯/异丙醇,30ml/100ml重结晶该产物混合物(三次)。产量:6.55g(94%)。1H NMR(DMSO-d6,360MHz)δ1.17(d,CH3CHSSC6H5,3H);3.24(CH3O-PEG,3H);3.40(s,PEG,180H);4.03(br t,mPEG-CH2,2H);5.28(S,C6H5CH2OCO,2H);7.45-8.35(m,C6H5)2,8H)。
将mPEG-MeDTB-碳酸硝基苯基酯(766mg,0.14mmol,1.29eq)溶于CHCl3(5ml)。将DSPE(70mg,0.093mol)和TEA(58.5μl,0.42mmol,4.5eq)加到PEG-溶液,并在50℃(油浴温度)下搅拌。20分钟后,TLC表明反应未完全。加入更多的mPEG-MeDTB-碳酸硝基苯基酯(共计1239mg,0.23mmol,2.47eq)和1-羟基苯并三唑(HOBt)(25mg,0.19mmol,2eq)。20分钟后,TLC(CHCl3∶MeOH∶H2O=90∶18∶2,有钼和茚三酮)表明反应完全。蒸发溶剂。将产物混合物溶于温(42℃)乙酸乙酯。它是混浊的溶液(TEA盐沉淀)。过滤该溶液并蒸发溶剂。往产物混合物中加入MeOH和2g的C8二氧化硅。再次蒸发溶剂。将含有C8二氧化硅的产物加到柱顶,并用以下溶剂洗脱:MeOH∶H2O梯度(压力),MeOH∶H2O 30∶70,100ml;MeOH H2O 50∶50,100ml;MeOH H2O 70∶30,250ml(起始物质洗脱);MeOH H2O 75∶25=40ml;MeOH H2O 80∶20,200ml(洗脱产物);MeOH=100ml;CHCl3∶MeOH∶H2O=90∶18∶2,100ml;CHCl3∶MeOH H2O=75∶36∶6,100ml。合并含有纯净产物的部分并蒸发得到无色浓稠液体产物。向其加入叔丁醇(5ml)、冻干然后在P2O5上真空干燥得到白色蓬松固体产物(467mg,83%产率)。1H NMR(DMSO-d6,360MHz)δ0.83(d,2(CH3),3H);1.16(d,CH3CHSSC6H5,3H);1.21(s,28(CH2,56H);1.47(brm,CH2CH2CO,4H);2.23(2 x t,CH2CH2CO,4H);3.50(s,PEG,180H);4.04(br t,mPEG-CH2,2H);4.05(反式d,PO4CH2CHCH2,1H);4.24(顺式d,PO4CH2CHCH2,1H);4.97(s,C6H5CH2OCO-DSPE,2H);5.03(brs,(PO4CH2CH,1H);7.32(d,C6H5,2H);7.53(d,C6H5,2H);7.52(br s,mPEG-OCONH,1H)。MALDI-TOFMS产生以相同44Da间隔分隔的钟状离子的分布,对应于环氧乙烷重复单元。该缀合物和mPEG-硫醇(大部分断裂的二硫化物)的平均分子量为6376和5368Da(理论分子量6053和5305道尔顿)。
B.mPEG-乙基DTB-DSPE
mPEG-尿烷乙基(乙基)二硫代羰基甲醇盐(2g,0.90mmol)溶于干甲醇(8ml)。溶液开始时混浊,但5分钟后变为澄清溶液。将巯基苄基醇(265.2mg,1.79mmol,2eq)加到该PEG-溶液。室温下搅拌该反应混合物30小时。将乙醚(70ml)加到该反应溶液以沉淀出产物,在4℃下保持过夜。过滤该白色固体并用乙酸乙酯/乙醚,30ml/70ml重结晶。产量:1.96g,(94%)。1H NMR(DMSO-d6,360MHz)δ0.86(d,CH3CH2CHSSC6H5CH2OH,3H);1.42(p,CH3CH2CHSSC6H5CH2OH,1H);1.64(p,CH3CH2CHSSC6H5CH2OH,1H);3.51(s,PEG,180H);4.03(t,mPEG-CH2,2H);4.47(d,HOCH2C6H5,2H);5.20(t,HOCH2C6H5,1H);7.31(d,C6H5,2H);7.42(br t,mPEG-OCONH,1H);7.49(d,C6H5,2H)。
在50℃(油浴温度)下将N-羟基-s-降冰片烯-2,3-二羧酸亚胺(HONB)(48mg,0.269mmol)加到CHCl3(3ml)中的DSPE(55mg,0.073mmol)。3-4分钟后,它变为澄清溶液。然后加入mPEG-EtDTB-氯甲酸硝基苯基酯(334mg,0.134mmol),随后加入三乙胺(TEA,45μl,0.329mmol)。20分钟后,TLC(CHCl3∶MeOH∶H2O=90∶18∶2)表明反应完全(钼和茚三酮喷雾)。蒸发溶剂。将产物混合物溶于甲醇,与C8二氧化硅(1g)混合,并通过旋转蒸发除去溶剂。将固体残留物加到C8-柱顶,然后用以下溶剂洗脱:MeOH∶H2O梯度(压力),MeOH∶H2O=30∶70,60ml;MeOH∶H2O=50∶50,60ml;MeOH∶H2O=70∶30,140ml;MeOH∶H2O=75∶25=140ml(起始物质洗脱);MeOH-H2O=80∶20,80ml;MeOH∶H2O=90∶10,140ml(洗脱产物);MeOH=40ml;CHCl3∶MeOH∶H2O=90∶18∶10,40ml。合并含有纯产物的部分并蒸发得到无色浓稠液体产物。加入叔丁醇(5ml)、冻干、然后在P2O5上真空干燥得到白色蓬松固体产物(175mg,78%产率)。1HNMR(DMSO-d6,360MHz)δ0.85(d,2(CH3),6H;d,CH3CHSSC6H5,3H);1.22(s,28(CH2),56H);1.49(br m,CH2CH2CO,4H);2.24(2 x t,CH2CH2CO,4H);3.50(s,PEG,180H);4.04(br t,mPEG-CH2,2H);4.08(反式d,PO4CH2CHCH2,1H);4.27(顺式d,PO4CH2CHCH2,1H);4.98(s,C6H5CH2OCO-DSPE,2H);5.06(br s,(PO4CH2CH,1H);7.34(d,C6H5,2H);7.53(d,C6H5,2H);7.55(br s,mPEG-OCONH,1H)。
实施例3
合成mPEG-DTB-氯甲酸硝基苯基酯
A.合成1-(巯基甲基)乙基氯化铵的方法
1. 2-氨基-1-甲基乙基硫酸氢酯。在冰浴中剧烈搅拌1-氨基-2-丙醇(22.53g,0.3mol)。在1小时内非常缓慢地加入硫酸(16.10ml,0.3mol)。在烧瓶中形成浓蒸汽和非常稠的溶液。在完成加入后,在170℃~180℃下与室内真空连接进行减压加热。在加热时,反应物变浅褐色。在除去所有的水后(大约1小时),冷却至室温。在冷却时形成褐色、玻璃状固体,它在用甲醇研制时可结晶。在60℃的水(50ml)中溶解它。加入足量的温甲醇而制得80%甲醇溶液。在冷却时,形成晶体,然后过滤晶体并在P2O5上干燥。产量:17.17g(37%)。1HNMR(D6-DMSO):δ1.16(d,CH3,3H);δ2.78(dd,NH3-CH2,1H);δ2.97(dd,NH3-CH2,1H);δ4.41(m,CH-OSO3,1H);δ7.69(s,H3N,3H)。熔点:248°-250℃(lit:250℃)
2. 5-甲基噻唑烷-2-硫酮。在250ml圆底烧瓶内在50%含水乙醇(40ml)中搅拌2-氨基-1-甲基乙基硫酸氢酯(23.03g,148mmol)和二硫化碳(10.71ml,178mmol,1.2eq.)。向其非常缓慢地滴加入在50%含水乙醇(50ml)中的氢氧化钠(13.06g,327mmol,2.2eq.)。在加入氢氧化钠时,将所有的起始物质溶解,溶液变橙色。回流反应物(85℃)40分钟,此后溶液变为亮黄色并形成浓稠沉淀。蒸发乙醇,然后加热该含水溶液,随后通过布氏漏斗过滤除去所有水溶性杂质。将残留的结晶溶于温乙醇,然后加入温水直至溶液含80%的水。将混合物冷却,然后离心,得到长针状晶体。产量:14.64g(75%)。1H NMR(D6-DMSO):δ1.33(d,CH3,3H);δ3.50(m,R3CH,1H);δ3.95(dd,N-CH2,1H);δ4.05(m,N-CH2,1H);δ10.05(s,NH,1H)。熔点:92.5-93.5(lit:94-95)。
3. 1-(巯基甲基)乙基氯化铵。将5-甲基噻唑烷-2-硫酮(6.5g,49mmol)置于250ml圆底烧瓶。加入含水盐酸溶液(40ml,18%在H2O中)并在油浴中加热该烧瓶。将反应物回流(120℃)一周。在整周内三次加入1ml浓盐酸。采用以乙酸乙酯作为洗脱剂的TLC监测反应。采用UV、茚三酮和碘蒸汽可视检验。在几乎整周的时间反应物为不均匀的混合物,其中油状起始物质比水浓。一周后油状起始物质消失,虽然在TLC仍可见。不再加热反应物并将其冷却至室温,然后冷冻至结晶起始物质。过滤结晶的起始物质。蒸发滤液并在P2O5和NaOH上干燥除去所有的水和HCl。用两部分的二乙醚(每次50ml)洗涤粗产物以除去所有的起始物质。再次在P2O5上干燥。产量:2.83g(45%)。1H NMR(D6 DMSO):δ1.33(d,CH3,3H);δ2.92(m,N-CH2,2H);δ3.12(m,SH,1H);δ3.18(m,R3-CH,1H);δ8.23(bs,NH3,3H)。熔点:80-82℃(lit:92-94)。
反应方案如图5所示。
B.合成mPEG-乙基-DTB-氯甲酸硝基苯基酯
1. 2-氨基-1-乙基乙基硫酸氢酯。在100ml的圆底烧瓶中在冰浴内剧烈搅拌1-氨基-2-丁醇(15ml,158mmol)。在1小时内非常缓慢地加入硫酸(8.43ml,158mmol)。在该烧瓶中形成浓蒸汽和非常稠的溶液。在添加完成后,在170和180℃之间在连接室内真空的减压下加热反应物。在加热时反应物变浅褐色。除去所有的水后(大约1小时),将其冷却至室温。在冷却时形成褐色、玻璃状固体。将其溶于热水(50ml),然后将其冷冻放置过夜。在冷却时,形成结晶然后过滤并在P2O5上干燥。产量:9.98g(37%)。1H NMR(D6-DMSO):δ0.87(t,CH3,3H);δ1.51(q,CH3-CH2,2H);δ2.82(dd,NH3-CH2,1H);δ3.00(dd,NH3-CH2,1H);δ4.21(m,CH-OSO3,1H);δ7.70(s,H3N,3H)。
2. 5-乙基噻唑烷-2-硫酮。在100ml圆底烧瓶内在50%含水乙醇(15ml)中搅拌2-氨基-1-乙基-乙基硫酸氢酯(9.98g,59mmol)和二硫化碳(4.26ml,71mmol,1.2eq.)。向其非常缓慢地滴加在50%含水乙醇(20ml)中的氢氧化钠(5.20g,130mmol,2.2eq.)。在加入氢氧化钠时,将所有的起始物质溶解,溶液变橙色。将反应物回流(85℃)40分钟,随后溶液变为亮黄并形成浓稠沉淀。蒸发乙醇,然后加热含水溶液,随后通过布氏漏斗过滤除去所有水溶性杂质。将残留晶体溶于温乙醇,然后加入温水直至溶液含80%水。将混合物冷却然后离心,得到针状晶体。产量:7.28g(86%)。1H NMR(D6-DMSO):δ0.88(t,CH3,3H);δ1.66(in,CH3-CH2,2H);δ3.58(m,R3CH,1H);δ3.93(m,N-CH2,2H);δ10.06(s,NH,1H)。熔点:76-78°(lit:76.6-76.9)。
3. 1-(巯基乙基)乙基氯化铵。将5-乙基噻唑烷-2-硫酮(7.24g,50mmol)置于250ml圆底烧瓶。加入含水盐酸溶液(45ml,18%在H2O中)并在油浴中加热烧瓶。在加热时,起始物质熔化,形成不均匀的混合物。将反应物回流(120℃)一周。在整周内四次加入1ml浓盐酸。采用以乙酸乙酯作为洗脱剂的TLC监测反应。采用UV、茚三酮和碘蒸汽可视检验。在整周的时间反应物为不均匀的混合物,其中油状起始物质比水浓。不再加热反应物并将其冷却至室温,然后冷冻至结晶起始物质。过滤结晶的起始物质。蒸发滤液并在P2O5和NaOH上干燥除去所有的水和HCl。用两部分的二乙醚(每次50ml)洗涤粗产物以除去所有的起始物质。再次在P2O5上干燥。产量:3.66g(52%).1H NMR(D6-DMSO):
该反应方案如图5所示。
实施例4
合成mPEG-DTB-脂质
将1,2-二硬脂酰-sn-甘油(500mg,0.8mmol)与苯共沸干燥(3次)。将氯甲酸对硝基苯基酯(242mg,1.2mmol,1.5eq)、二甲基氨基吡啶(DMAP)(10mg,0.08mmol,0.1eq)和TEA(334.5μl,2.4mmol,3eq)加到在CHCl3(5ml)中的1,2-二硬脂酰甘油。室温下将反应混合物搅拌2h。TLC(甲苯∶乙酸乙酯=7∶3)表明反应完全。然后用10%柠檬酸提取产物混合物以除去二甲基氨基吡啶(DMAP),用乙腈(3ml,4次)洗涤以除去过量的氯甲酸对硝基苯基酯。在P2O5上真空干燥纯产物。产量:557mg(88%).%)。1H NMR(CHCl3,360MHz)δ0.88(t,end CH3,6H);1.25(s,28xCH2,56H);1.58(m,CH2CH2CO,4H);2.34(2xt,CH2CO,4H);4.22(反式d,CH2OCOC17H35,1H);4.35(m,OCOOCH2CH,2H);4.51(顺式d,CH2OCOC17H35,1H);5.37(m,OCOOCH2CH,1H);7.39(d,C6H5,2H);8.28(d,C6H5,2H)。
将乙二胺(42μl,0.63mmol,5倍过量)和吡啶(200μl)加到CHCl3(1ml)。将2-二硬脂酰-sn-碳酸对硝基苯基酯(100mg,0.13mmol)溶于CHCl3(1ml)并在0℃(冰水)下用巴斯德移液管滴加到乙二胺溶液并持续过夜(16小时)。TLC(CHCl3∶MeOH∶H2O 90∶18∶2,和CHCl3∶MeOH=90∶10)表明反应完全。蒸发溶剂除去吡啶。然后将产物混合物溶于CHCl3,装载到柱上(Aldrich,硅胶,60°A,200-400目),并用以下溶剂洗脱:CHCl3∶CH3COCH3,和CHCl3∶MeOH梯度,CHCl3∶CH3COCH3=90∶10,60ml(upper spot eluted);CHCl3∶NeOH=90∶10,60ml(洗脱产物)。合并含有纯产物的部分并蒸发。加入叔丁醇并在P2O5上真空干燥。产量:64mg(75%)。1H NMR(DMSO-d6,360MHz)δ.83(t,终点CH3,6H);1.22(s,28xCH2,56H);1.51(m,CH2CH2CO,4H);2.25(2xt,CH2CO,4H);2.83(m,H2NCH2CH2NH,2H);3.21(m,H2NCH2CH2NH,2H);4.10-4.14(m & 顺式d,COOCH2CHCH2,4H);5.17(m,OCOOCH2CH,1H);7.78(m,H2NCH2CH2NH,2H)。
将mPEG-MeDTB-氯甲酸硝基苯基酯(400mg,0.162mmol,2.2eq)溶于CHCl3in(2ml)。将1,2-硬脂酰基-sn-亚乙胺(51mg,0.075mmol)和TEA(37μl,0.264mmol,3.52eq)加到溶液。然后在45℃下将反应混合物搅拌20分钟。TLC(CHCl3∶MeOH∶H2O=90∶18∶2,和CHCl3∶MeOH=90∶10)表明反应完全。蒸发溶剂。将产物混合物溶于甲醇。加入2g的C8二氧化硅然后蒸发溶剂。将含有产物混合物的C8二氧化硅加到C8柱顶((Supelco,Supel clean.Lot no.SP0824),用以下溶剂洗脱:MeOH∶H2O梯度(压力)、MeOH∶H2O=60∶40,40ml、MeOH∶H2O=70∶30,80ml(起始物质洗脱)、MeOH∶H2O=80∶20,40ml、MeOH∶H2O=90∶10=20ml、CHCl3∶MeOH∶H2O=5∶80∶15,20ml、CHCl3∶MeOH∶H2O=90∶18∶10,40ml(产物洗脱)。合并含有纯产物的部分并蒸发得到无色浓稠液体产物。加入叔丁醇(5ml)并冻干溶液,然后在P2O5上真空干燥得到白色固体产物(200mg,89%产率)。1H NMR(DMSO-d6,360MHz)δδ.83(t,endCH3,6H);1.22(s,28xCH2,56H);1.48(m,CH2CH2CO,4H);2.25(2 x t,CH2CO,4H);3.10(m,HNCH2CH2NH,4H);3.50(s,PEG,180H);4.04(t,mPEG-CH2,2H);4.09(反式d,COOCH2CHCH2,1H);4.25(顺式d,COOCH2CHCH2,1H);4.98(s,C6H5CH2OCO,2H);5.23(m,COOCH2CHCH2,1H);7.18(m,NHCH2CH2NH,2H);7.33(d,C6H5,2H);7.38(m,mPEG-OCONH,1H);7.52(d,C6H5,2H)。
该反应方案如图6A所示。
实施例5
体外断裂mPEG-DTB-DSPE化合物
将邻mPEG-DTB-DSPE和对mPEG-DTB-DSPE(如实施例1所述制备)在存在和不存在150μM半胱氨酸下加到缓冲的含水溶液(pH7.2)。HPLC(Phenomenex C8 Prodigy,4.6×50mm柱,在277nm处检测,流动相甲醇/水95∶5和0.1%三氟乙酸,流速1mL/min)监测到缀合物消失。结果如图7A所示,其中邻缀合物由空心圆表示而对缀合物由空心正方形表示。
实施例6
体外断裂脂质体中的邻和对mPEG-DTB-DSPE化合物
A.脂质体制备
将部分氢化的脂质磷脂酰胆碱(PHPC)、胆固醇和邻或对mPEG-DTB-DSPE(如实施例1所述制备,mPEG MW=1980道尔顿)以95∶5∶3的摩尔比率分别溶于适宜的有机溶剂,一般为1∶1或1∶3比率的氯仿/甲醇。通过旋转蒸发除去溶剂形成干燥的脂质薄膜。用含水缓冲剂水合该薄膜,通过挤压到120nm平均直径的大小形成脂质体。
B.体外表征
在37℃、pH7.2、含有5mM EDTA的磷酸盐缓冲盐水中,在150μM半胱氨酸的存在下温育脂质体。HPLC(Phenomenex C8 Prodigy,4.6×50mm柱,在277nm处检测,流动相甲醇/水95∶5和0.1%三氟乙酸,流速1mL/min)监测到缀合物消失。结果如图7B所示,其中含有邻缀合物的脂质体由实心圆表示而含对缀合物的脂质体由实心正方形表示。空心圆和空心正方形对应于胶束形式的邻mPEG-DTB-DSPE和对mPEG-DTB-DSPE(如以上在实施例5所讨论,图7A)。
实施例7
体外断裂脂质体中的邻和对mPEG-DTB-DSPE化合物
A.脂质体制备
将脂质二油酰磷脂酰乙醇胺(DOPE)和邻或对mPEG-DTB-DSPE(如实施例1所述制备,mPEG Mw=1980道尔顿)以97∶3的摩尔比率溶于氯仿/甲醇1∶1。通过旋转蒸发除去溶剂形成干燥的脂质薄膜。用含有各30mM的每种荧光团对二甲苯-双-溴化吡啶鎓和8-羟基芘三磺酸三钠的含水溶液水合该脂质薄膜。用含水缓冲剂水合该薄膜,通过挤压到100nm平均直径的大小形成脂质体。
B.体外表征
在37℃、pH7.2的HEPES缓冲剂中,在15μM、150μM、300μM和1.5mM浓度的半胱氨酸的存在下温育脂质体。测定染料的释放百分率作为高于预温育的试样(零释放)的试样荧光(λem=512nm,λem=413nm-pH-独立性等消光点)的增长,将这些试样归一化为在用0.2%的Triton X-100溶解预温育的试样后得到的荧光增长(100%释放)(Kirpotin,D.等人,FEBS Letters,388:115-118(1996))。各种半胱氨酸浓度下的含邻位化合物脂质体的结果如图8A所示,含对位化合物的脂质体的结果如图8B所示。
实施例8
体内表征含有mPEG-DTB-DSPE化合物的脂质体
A.脂质体制备
将部分氢化的脂质磷脂酰胆碱(PHPC)、胆固醇和对mPEG-DTB-DSPE(如实施例1所述制备,mPEG MW=1980道尔顿)以55∶40∶5的摩尔百分比率分别溶于有机溶剂。通过旋转蒸发除去溶剂形成干燥的脂质薄膜。用含有二亚乙基三胺五乙酸(EDTA)的含水缓冲剂水合该薄膜形成脂质体。将脂质体小型化至平均直径为120nm,然后除去未截留的EDTA并往外部介质中加入In111。将脂质体温育充足的时间以使In111穿过脂质双层和与EDTA螯合。
B.体内给药
将小鼠分成两个研究小组。将上述的脂质体组合物注射进所有的受试动物。一组受试动物还在脂质体注射后的1、3和5小时接受200μL注射的200mM半胱氨酸。另一受试组在相同的时间点接受注射盐水。通过监测血样中的In111来测定血液中脂质体的含量。结果如图10所示。
虽然已描述了本发明的特定实施方案,但对本领域技术人员来说显然可以在不背离本发明的情况下进行各种改变和修正。
Claims (41)
1.具有以下一般结构的化合物:其中R1为含有用于连接到二硫代苄基部分的键的亲水性聚合物;R2选自H、烷基和芳基;R3选自O(C=O)R4、S(C=O)R4和O(C=S)R4;R4包括含胺配体;而R5选自H、烷基和芳基;其中CH2-R3的取向选自邻位和对位。
2.权利要求1的化合物,其中R5为H而R2选自CH3、C2H5和C3H8。
3.根据权利要求1或2的化合物,其中该含胺配体R4选自多肽、含胺药物和含胺脂质。
4.根据权利要求1或2的化合物,其中该含胺配体R4为包括单烃尾或双烃尾的含胺脂质。
5.权利要求4的化合物,其中该含胺脂质为具有双烃尾的磷脂。
6.根据权利要求1、3、4或5任一项的化合物,其中R2和R5为烷基。
7.根据前述权利要求任一项的化合物,其中R1选自聚乙烯吡咯烷酮、聚乙烯基甲基醚、聚甲基噁唑啉、聚乙基噁唑啉、聚羟基丙基噁唑啉、聚羟基丙基-甲基丙烯酰胺、聚甲基丙烯酰胺、聚二甲基-丙烯酰胺、聚羟基丙基甲基丙烯酸酯、聚羟基乙基丙烯酸酯、羟甲基纤维素、羟乙基纤维素、聚乙二醇、聚天冬酰胺、它们的共聚物,以及聚环氧乙烷-聚环氧丙烷。
8.根据权利要求1-6任一项的化合物,其中R1为聚乙二醇。
9.权利要求8的化合物,其中R5为H而R2为CH3或C2H5。
10.含有前述权利要求任一项的化合物的脂质体。
11.根据权利要求1、2、7、8或9任一项的化合物,其中该含胺配体R4为多肽。
12.权利要求11的化合物,其中该多肽为重组多肽。
13.权利要求11的化合物,其中该多肽为细胞因子。
14.权利要求11的化合物,其中该多肽选自干扰素、白介素、生长因子和酶。
16.权利要求15的组合物,其中R2选自CH3、C2H5和C3H8。
17.权利要求15的组合物,其中R3为O(C=O)R4而R4为含羟基或含氧的离去基团。
18.权利要求15的组合物,其中该离去基团来源于选自以下的化合物:氯化物、对硝基苯酚、邻硝基苯酚、N-羟基-四氢邻苯二甲酰亚胺、N-羟基琥珀酰亚胺、N-羟基戊二酰亚胺、N-羟基降冰片烯-2,3-二羧基亚胺、1-羟基苯并三唑、3-羟基吡啶、4-羟基吡啶、2-羟基吡啶、1-羟基-6-三氟甲基苯并三唑、咪唑、三唑、N-甲基-咪唑、五氟苯酚、三氟苯酚和三氯苯酚。
19.权利要求15的组合物,其中该化合物与代替R4的含胺配体反应形成包括该含胺配体的缀合物。
20.权利要求19的组合物,其中该含胺配体包括磷脂。
21.权利要求19的组合物,其中该含胺配体包括多肽。
22.根据权利要求20或21的组合物,其中R1选自聚乙烯吡咯烷酮、聚乙烯基甲基醚、聚甲基噁唑啉、聚乙基噁唑啉、聚羟基丙基噁唑啉、聚羟基丙基-甲基丙烯酰胺、聚甲基丙烯酰胺、聚二甲基丙烯酰胺、聚羟基丙基甲基丙烯酸酯、聚羟基乙基丙烯酸酯、羟甲基纤维素、羟乙基纤维素、聚乙二醇、聚天冬酰胺、它们的共聚物,以及聚环氧乙烷-聚环氧丙烷。
23.根据权利要求20或21的组合物,其中R1包括聚乙二醇。
24.根据权利要求23的组合物,其中R2为CH3或C2H5。
25.权利要求20的组合物,其中含有该缀合物的组合物包括脂质体。
26.权利要求25的组合物,其中该脂质体进一步包括截留治疗剂。
27.权利要求21的组合物,其中该多肽包括重组多肽。
28.权利要求21的组合物,其中该多肽包括细胞因子。
29.权利要求21的组合物,其中该多肽选自干扰素、白介素、生长因子和酶。
31.权利要求30的组合物,其中R5为H而R2选自CH3,C2H5和C3H8。
32.根据权利要求30或31的组合物,其中该含胺脂质包括磷脂。
33.根据权利要求30-32任一项的组合物,其中其中R1选自聚乙烯吡咯烷酮、聚乙烯基甲基醚、聚甲基噁唑啉、聚乙基噁唑啉、聚羟基丙基噁唑啉、聚羟基丙基-甲基丙烯酰胺、聚甲基丙烯酰胺、聚二甲基-丙烯酰胺、聚羟基丙基甲基丙烯酸酯、聚羟基乙基丙烯酸酯、羟甲基纤维素、羟乙基纤维素、聚乙二醇、聚天冬酰胺、它们的共聚物,以及聚环氧乙烷-聚环氧丙烷。
34.根据权利要求30-32任一项的组合物,其中R1包括聚乙二醇。
35.根据权利要求30-34任一项的组合物,其中该脂质体进一步包括截留治疗剂。
36.一种改善具有可释放亲水性聚合物链的表面包被的脂质体的血液循环寿命的方法,包括制备含约1%-约20%的具有以下一般结构的化合物的脂质体:其中R1为含有用于连接到二硫代苄基部分的键的亲水性聚合物;R2选自H、烷基和芳基;R3选自O(C=O)R4、S(C=O)R4和O(C=S)R4;R4包括含胺脂质;而R5选自H、烷基和芳基;其中CH2-R3的取向选自邻位和对位。
37.权利要求36的方法,其中R5为H而R2选自CH3、C2H5和C3H8。
38.根据权利要求36或37的方法,其中该含胺脂质包括磷脂。
39.根据权利要求36-38任一项的方法,其中R1选自聚乙烯吡咯烷酮、聚乙烯基甲基醚、聚甲基噁唑啉、聚乙基噁唑啉、聚羟基丙基噁唑啉、聚羟基丙基-甲基丙烯酰胺、聚甲基丙烯酰胺、聚二甲基-丙烯酰胺、聚羟基丙基甲基丙烯酸酯、聚羟基乙基丙烯酸酯、羟甲基纤维素、羟乙基纤维素、聚乙二醇、聚天冬酰胺、它们的共聚物,以及聚环氧乙烷-聚环氧丙烷。
40.根据权利要求36-38任一项的方法,其中R1包括聚乙二醇。
41.根据权利要求36-40任一项的方法,其中该脂质体进一步包括截留治疗剂。
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN106519221A (zh) * | 2015-09-10 | 2017-03-22 | 中国科学院高能物理研究所 | 一种聚乙二醇/聚酰胺胺共聚物、其制备方法及包含该共聚物的两亲性siRNA载体 |
CN106519211A (zh) * | 2015-09-10 | 2017-03-22 | 中国科学院高能物理研究所 | 一种两亲性聚合物以及由其形成的磁性胶束纳米载体和其用途 |
Families Citing this family (92)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5981501A (en) * | 1995-06-07 | 1999-11-09 | Inex Pharmaceuticals Corp. | Methods for encapsulating plasmids in lipid bilayers |
US7422902B1 (en) * | 1995-06-07 | 2008-09-09 | The University Of British Columbia | Lipid-nucleic acid particles prepared via a hydrophobic lipid-nucleic acid complex intermediate and use for gene transfer |
WO1998051278A2 (en) | 1997-05-14 | 1998-11-19 | Inex Pharmaceuticals Corporation | High efficiency encapsulation of charged therapeutic agents in lipid vesicles |
US7238368B2 (en) * | 1999-04-23 | 2007-07-03 | Alza Corporation | Releasable linkage and compositions containing same |
US7303760B2 (en) * | 1999-04-23 | 2007-12-04 | Alza Corporation | Method for treating multi-drug resistant tumors |
US7112337B2 (en) * | 1999-04-23 | 2006-09-26 | Alza Corporation | Liposome composition for delivery of nucleic acid |
EP1880736A1 (en) * | 1999-04-23 | 2008-01-23 | Alza Corporation | Releasable linkage and composition containing same |
GB9915074D0 (en) * | 1999-06-28 | 1999-08-25 | Cortecs Plc | Ligand-binding composition |
US7094423B1 (en) * | 1999-07-15 | 2006-08-22 | Inex Pharmaceuticals Corp. | Methods for preparation of lipid-encapsulated therapeutic agents |
WO2002076428A1 (en) * | 2001-03-26 | 2002-10-03 | Alza Corporation | Liposome composition for improved intracellular delivery of a therapeutic agent |
DE60231868D1 (de) | 2001-04-24 | 2009-05-20 | Purdue Research Foundation | Folat-mimetika und deren folatrezeptorbindende konjugate |
EP1441763A2 (en) * | 2001-11-07 | 2004-08-04 | Inex Pharmaceuticals Corp. | Mucosal adjuvants comprising an oligonucleotide and a cationic lipid |
US7842498B2 (en) * | 2001-11-08 | 2010-11-30 | Bio-Rad Laboratories, Inc. | Hydrophobic surface chip |
US7211440B2 (en) | 2002-03-08 | 2007-05-01 | Wallac Oy | Dissociative fluorescence enhancement assay |
US6737524B2 (en) * | 2002-03-25 | 2004-05-18 | Paul K. Smith | Activated polyethylene glycol compounds |
US20040009944A1 (en) * | 2002-05-10 | 2004-01-15 | Inex Pharmaceuticals Corporation | Methylated immunostimulatory oligonucleotides and methods of using the same |
CA2752143C (en) | 2002-05-15 | 2014-09-23 | Sutter West Bay Hospitals | Delivery of nucleic acid-like compounds |
WO2003097647A1 (en) * | 2002-05-15 | 2003-11-27 | Endocyte, Inc. | Vitamin-mitomycin conjugates |
EP1581186A2 (en) * | 2002-12-03 | 2005-10-05 | Blanchette Rockefeller Neurosciences Institute | Artificial low-density lipoprotein carriers for transport of substances across the blood-brain barrier |
DK1592457T3 (da) | 2003-01-27 | 2012-10-22 | Endocyte Inc | Folat-vinblastin-konjugat som lægemiddel |
WO2005034979A2 (en) * | 2003-10-11 | 2005-04-21 | Inex Pharmaceuticals Corporation | Methods and compositions for enhancing innate immunity and antibody dependent cellular cytotoxicity |
US20050191344A1 (en) * | 2004-01-15 | 2005-09-01 | Samuel Zalipsky | Liposome composition for delivery of therapeutic agents |
US7282590B2 (en) * | 2004-02-12 | 2007-10-16 | The Research Foundation Of State University Of New York | Drug conjugates |
US7931693B2 (en) * | 2004-02-26 | 2011-04-26 | Endosphere, Inc. | Method and apparatus for reducing obesity |
US9592277B2 (en) * | 2004-04-14 | 2017-03-14 | Avirid, Inc. | Compositions with modified nucleases targeted to viral nucleic acids and methods of use for prevention and treatment of viral diseases |
AU2005238015A1 (en) * | 2004-04-21 | 2005-11-10 | Alza Corporation | Polymer conjugate releasable under mild thiolytic conditions |
JP2007533750A (ja) * | 2004-04-21 | 2007-11-22 | アルザ コーポレイション | 緩和なチオール分解条件下で遊離可能なポリマー結合体 |
JP5149620B2 (ja) | 2004-07-23 | 2013-02-20 | エンドサイト,インコーポレイテッド | 2価リンカーおよびその結合体 |
CA2577786A1 (en) * | 2004-09-03 | 2006-03-16 | Alza Corporation | Endogenously-formed conjugate of albumin |
TW200612993A (en) * | 2004-10-08 | 2006-05-01 | Alza Corp | Lipopolymer conjugates |
AU2005302255A1 (en) * | 2004-10-28 | 2006-05-11 | Alza Corporation | Lyophilized liposome formulations and method |
US8044200B2 (en) * | 2005-03-16 | 2011-10-25 | Endocyte, Inc. | Synthesis and purification of pteroic acid and conjugates thereof |
AU2006279304A1 (en) * | 2005-08-19 | 2007-02-22 | Endocyte, Inc. | Multi-drug ligand conjugates |
EP2382995A3 (en) * | 2005-08-19 | 2013-09-25 | Endocyte, Inc. | Ligand conjugates of Vinca alkaloids, analogs and derivatives |
KR20090034385A (ko) * | 2006-07-19 | 2009-04-07 | 더 보드 오브 리전츠 오브 더 유니버시티 오브 텍사스 시스템 | 염증성 장질환을 치료하기 위한 5-아미노살리실산 함유 약제 및 인지질의 제제 |
US20100329664A1 (en) * | 2007-01-23 | 2010-12-30 | Lim Dae-Soon | Shutter device for camera |
WO2008101231A2 (en) * | 2007-02-16 | 2008-08-21 | Endocyte, Inc. | Methods and compositions for treating and diagnosing kidney disease |
WO2008112873A2 (en) * | 2007-03-14 | 2008-09-18 | Endocyte, Inc. | Binding ligand linked drug delivery conjugates of tubulysins |
ITRM20070327A1 (it) * | 2007-06-11 | 2008-12-12 | Univ Palermo | Vettori colloidali a struttura poliamminoacidica per il rilascio orale di peptidi e proteine e relativo metodo di produzione. |
CN104383553A (zh) * | 2007-06-25 | 2015-03-04 | 恩多塞特公司 | 含有亲水性间隔区接头的共轭物 |
US9877965B2 (en) | 2007-06-25 | 2018-01-30 | Endocyte, Inc. | Vitamin receptor drug delivery conjugates for treating inflammation |
US20100210575A1 (en) * | 2007-06-29 | 2010-08-19 | Wisconsin Alumni Research Foundation | Structuring effect of cholesterol in peg-phospholipid micelles, drug delivery of amphotericin b, and combination antifungals |
CN101854917A (zh) * | 2007-07-20 | 2010-10-06 | 巴斯夫欧洲公司 | 包含跨膜转运触发体系的小泡 |
US8440787B2 (en) | 2007-10-23 | 2013-05-14 | Nektar Therapeutics | Hydroxyapatite-targeting multiarm polymers and conjugates made therefrom |
EP2209374B1 (en) | 2007-10-25 | 2014-12-03 | Endocyte, Inc. | Tubulysins and processes for preparing |
US9259398B1 (en) * | 2007-11-26 | 2016-02-16 | Abbott Cardiovascular Systems Inc. | Bioactive agent-loaded targeting micelles |
CA2725535C (en) * | 2008-05-23 | 2016-01-05 | The University Of British Columbia | Modified drugs for use in liposomal nanoparticles |
CN109293927A (zh) | 2009-02-04 | 2019-02-01 | 布里格姆及妇女医院股份有限公司 | 纳米级铂化合物及其使用方法 |
MX2011012597A (es) | 2009-05-27 | 2012-04-19 | Selecta Biosciences Inc | Nanoportadores que poseen componentes con diferentes tasas de liberacion. |
KR101873179B1 (ko) * | 2009-08-26 | 2018-06-29 | 셀렉타 바이오사이언시즈, 인크. | T-세포 도움을 유도하는 조성물 |
WO2011051916A2 (en) * | 2009-11-02 | 2011-05-05 | Universite De Geneve | Stabilized protein formulations and use thereof |
NZ716192A (en) | 2009-12-01 | 2017-07-28 | Shire Human Genetic Therapies | Delivery of mrna for the augmentation of proteins and enzymes in human genetic diseases |
WO2011119995A2 (en) | 2010-03-26 | 2011-09-29 | Cerulean Pharma Inc. | Formulations and methods of use |
DE102010042338A1 (de) * | 2010-10-12 | 2012-04-12 | Bayer Technology Services Gmbh | Zusammensetzung zur Behandlung von Krebs mit kontrollierter Freisetzung des Wirkstoffes |
EP2640190A4 (en) | 2010-11-05 | 2016-05-11 | Selecta Biosciences Inc | MODIFIED NICOTINIC COMPOUNDS AND ASSOCIATED METHODS |
CA2834619A1 (en) | 2011-04-29 | 2012-11-01 | Selecta Biosciences, Inc. | Controlled release of immunosuppressants from synthetic nanocarriers |
DK2717893T3 (da) | 2011-06-08 | 2019-07-22 | Translate Bio Inc | Lipid nanopartikelsammensætninger og fremgangsmåder til mrna-levering |
EP2718269B1 (en) * | 2011-06-08 | 2018-01-31 | Translate Bio, Inc. | Cleavable lipids |
EP2606884A1 (en) | 2011-12-21 | 2013-06-26 | Ecole Polytechnique Fédérale de Lausanne (EPFL) | Inhibitors of notch signaling pathway and use thereof in treatment of cancers |
US10080805B2 (en) | 2012-02-24 | 2018-09-25 | Purdue Research Foundation | Cholecystokinin B receptor targeting for imaging and therapy |
EP3865123A1 (en) | 2012-03-29 | 2021-08-18 | Translate Bio, Inc. | Lipid-derived neutral nanoparticles |
US20140080175A1 (en) | 2012-03-29 | 2014-03-20 | Endocyte, Inc. | Processes for preparing tubulysin derivatives and conjugates thereof |
WO2013185067A1 (en) | 2012-06-08 | 2013-12-12 | Shire Human Genetic Therapies, Inc. | Nuclease resistant polynucleotides and uses thereof |
BR112015008365A2 (pt) | 2012-10-16 | 2017-07-04 | Endocyte Inc | composto da fórmula b-l(d)x, ou um sal farmaceuticamente aceitável do mesmo, composição farmacêutica, uso de um composto, composição de forma de dosagem unitária ou dose unitária, composição para tratar um câncer em um paciente, e método para tratar um câncer em um paciente |
DK3467108T3 (da) | 2013-03-14 | 2024-06-10 | Translate Bio Inc | Fremgangsmåder til oprensning af messenger-RNA |
UA117008C2 (uk) | 2013-03-14 | 2018-06-11 | Шир Хьюман Дженетік Терапіс, Інк. | IN VITRO ТРАНСКРИБОВАНА мРНК ТА КОМПОЗИЦІЯ, ЩО ЇЇ МІСТИТЬ, ДЛЯ ЗАСТОСУВАННЯ В ЛІКУВАННІ МУКОВІСЦИДОЗУ В ССАВЦЯ |
US9827552B2 (en) | 2013-07-17 | 2017-11-28 | Clemson University | Functionalized lipid modification of solid phase surfaces for use in chromatography |
EP3060303B1 (en) | 2013-10-22 | 2018-11-14 | Translate Bio, Inc. | Mrna therapy for argininosuccinate synthetase deficiency |
EP3060257B1 (en) | 2013-10-22 | 2021-02-24 | Translate Bio, Inc. | Lipid formulations for delivery of messenger rna |
EA201992208A1 (ru) | 2013-10-22 | 2020-07-31 | Транслейт Био, Инк. | ЛЕЧЕНИЕ ФЕНИЛКЕТОНУРИИ С ПРИМЕНЕНИЕМ мРНК |
JP6571679B2 (ja) | 2014-04-25 | 2019-09-04 | トランスレイト バイオ, インコーポレイテッド | メッセンジャーrnaの精製方法 |
JP6557722B2 (ja) | 2014-05-30 | 2019-08-07 | シャイアー ヒューマン ジェネティック セラピーズ インコーポレイテッド | 核酸の送達のための生分解性脂質 |
WO2015191576A1 (en) | 2014-06-09 | 2015-12-17 | Lipomedix Pharmaceuticals Ltd. | Combination therapy comprising a liposomal prodrug of mitomycin c and radiotherapy |
ES2964588T3 (es) | 2014-06-24 | 2024-04-08 | Translate Bio Inc | Composiciones enriquecidas estereoquímicamente para la administración de ácidos nucleicos |
EP3270898B1 (en) | 2015-03-17 | 2021-05-05 | Lipomedix Pharmaceuticals Ltd. | Methods for the treatment of bladder cancer |
CN105866311B (zh) * | 2016-05-25 | 2017-05-31 | 福建出入境检验检疫局检验检疫技术中心 | 测定鸡肉中抗病毒药物残留的uplc‑ms/ms方法 |
GB2552301A (en) * | 2016-07-11 | 2018-01-24 | Evox Therapeutics Ltd | Metabolic drug loading of EVs |
US20200079785A1 (en) | 2016-11-08 | 2020-03-12 | Mallinckrodt Llc | Mitomycin c prodrug liposome formulations and uses thereof |
CA3054062A1 (en) | 2017-02-27 | 2018-08-30 | Translate Bio, Inc. | Novel codon-optimized cftr mrna |
EP3624824B1 (en) | 2017-05-16 | 2024-07-10 | Translate Bio, Inc. | Codon-optimized mrna encoding cftr for use in treating cystic fibrosis |
EP3655039A1 (en) * | 2017-07-17 | 2020-05-27 | Technische Universiteit Eindhoven | Applicable chemical composition comprising an agent conjugated to a hydrophobic moiety and a carrier |
US10618896B2 (en) | 2017-08-22 | 2020-04-14 | Dynavax Technologies Corporation | Alkyl chain modified imidazoquinoline TLR7/8 agonist compounds and uses thereof |
JP2021503005A (ja) | 2017-11-14 | 2021-02-04 | ダイナバックス テクノロジーズ コーポレイション | Tlr7/8アゴニスト化合物の切断可能なコンジュゲート、その調製方法および使用 |
EA202190024A1 (ru) | 2018-06-21 | 2021-05-19 | Селлестия Биотек Аг | Способ получения амино-диариловых эфиров и гидрохлоридных солей амино-диариловых эфиров |
CN118421617A (zh) | 2018-08-24 | 2024-08-02 | 川斯勒佰尔公司 | 用于纯化信使rna的方法 |
WO2020144657A1 (en) | 2019-01-11 | 2020-07-16 | Lipomedix Pharmaceuticals Ltd. | Liposome composition comprising liposomal prodrug of mitomycin c and method of manufacture |
US20220169642A1 (en) | 2019-04-10 | 2022-06-02 | Cellestia Biotech Ag | Compounds for the treatment of oncovirus induced cancer and methods of use thereof |
EP4008324A1 (en) | 2020-12-07 | 2022-06-08 | Cellestia Biotech AG | Combinations comprising an inhibitor of an anti-apoptotic protein, such as bcl-2, bcl-xl, bclw or mcl-1, and a notch signaling pathway inhibitor for treating cancer |
US20240041884A1 (en) | 2020-12-07 | 2024-02-08 | Cellestia Biotech Ag | Pharmaceutical Combinations for Treating Cancer |
WO2022253794A2 (en) | 2021-06-02 | 2022-12-08 | Cellestia Biotech Ag | Method for treating an autoimmune and inflammatory disease |
EP4429661A1 (en) | 2021-11-08 | 2024-09-18 | Cellestia Biotech AG | Pharmaceutical combinations for treating cancer |
EP4223292A1 (en) | 2022-02-07 | 2023-08-09 | Cellestia Biotech AG | Pharmaceutical combinations for treating cancer |
Family Cites Families (38)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4179337A (en) | 1973-07-20 | 1979-12-18 | Davis Frank F | Non-immunogenic polypeptides |
JPS5240B2 (zh) | 1973-12-17 | 1977-01-05 | ||
GB8430252D0 (en) | 1984-11-30 | 1985-01-09 | Beecham Group Plc | Compounds |
US4917888A (en) | 1985-06-26 | 1990-04-17 | Cetus Corporation | Solubilization of immunotoxins for pharmaceutical compositions using polymer conjugation |
US5059421A (en) * | 1985-07-26 | 1991-10-22 | The Liposome Company, Inc. | Preparation of targeted liposome systems of a defined size distribution |
US4766106A (en) | 1985-06-26 | 1988-08-23 | Cetus Corporation | Solubilization of proteins for pharmaceutical compositions using polymer conjugation |
JPH0615532B2 (ja) * | 1986-02-03 | 1994-03-02 | テルモ株式会社 | 5−フルオロウラシル誘導体およびこれを含有する医薬製剤 |
US4766105A (en) * | 1986-10-31 | 1988-08-23 | Shell Oil Company | Ethylene oxide catalyst and process for preparing the catalyst |
JPH01113391A (ja) * | 1987-10-24 | 1989-05-02 | Kyowa Hakko Kogyo Co Ltd | マイトマイシン誘導体 |
US4952394A (en) * | 1987-11-23 | 1990-08-28 | Bristol-Myers Company | Drug-monoclonal antibody conjugates |
ZA886812B (en) * | 1987-11-23 | 1989-07-26 | Bristol Myers Co | Anti-tumor prodrugs |
US5103556A (en) | 1988-05-05 | 1992-04-14 | Circon Corporation | Method of manufacturing an electrohydraulic probe |
US4902502A (en) | 1989-01-23 | 1990-02-20 | Cetus Corporation | Preparation of a polymer/interleukin-2 conjugate |
US5585112A (en) * | 1989-12-22 | 1996-12-17 | Imarx Pharmaceutical Corp. | Method of preparing gas and gaseous precursor-filled microspheres |
TW211015B (zh) * | 1990-01-11 | 1993-08-11 | Nippon Shinyaku Co Ltd | |
US5169934A (en) * | 1990-05-14 | 1992-12-08 | Anergen, Inc. | Intracellularly cleavable compounds |
EP0706373B1 (en) | 1992-03-23 | 2000-07-19 | Georgetown University | Liposome encapsulated taxol and a method of using the same |
JP2813511B2 (ja) * | 1992-06-10 | 1998-10-22 | 大日本スクリーン製造株式会社 | ロールコータによる基板表面への塗液塗布方法 |
AU5092893A (en) | 1992-09-02 | 1994-03-29 | Georgetown University | Method of encapsulating anthracycline glycosides in liposomes |
US5395619A (en) | 1993-03-03 | 1995-03-07 | Liposome Technology, Inc. | Lipid-polymer conjugates and liposomes |
US5820873A (en) * | 1994-09-30 | 1998-10-13 | The University Of British Columbia | Polyethylene glycol modified ceramide lipids and liposome uses thereof |
EP0804459A4 (en) * | 1995-01-16 | 1999-05-26 | Commw Scient Ind Res Org | CONJUGATES THERAPEUTIC COMPOUND - FATTY ACID |
WO1996022302A1 (en) | 1995-01-20 | 1996-07-25 | Florida State University | Sex-specific dna probe for parrots, methods and kits |
JPH09268190A (ja) * | 1996-04-02 | 1997-10-14 | Sagami Chem Res Center | マイトマイシンc誘導体及び非受容体型チロシンキナーゼ阻害剤 |
JP2001503396A (ja) * | 1996-10-11 | 2001-03-13 | アルザ コーポレイション | 治療用リポソーム組成物および方法 |
TW520297B (en) * | 1996-10-11 | 2003-02-11 | Sequus Pharm Inc | Fusogenic liposome composition and method |
JPH1160499A (ja) | 1997-08-22 | 1999-03-02 | Hiroshi Maeda | 抗腫瘍剤 |
SE513149C2 (sv) * | 1997-12-05 | 2000-07-17 | Katarina Edwards | Läkemedelsdistributionssystem med tvåstegsmålsökning, till specifika celler eller vävnad och till dess cellkärna |
US6180095B1 (en) | 1997-12-17 | 2001-01-30 | Enzon, Inc. | Polymeric prodrugs of amino- and hydroxyl-containing bioactive agents |
ES2224649T3 (es) | 1998-04-28 | 2005-03-01 | Applied Research Systems Ars Holding N.V. | Conjugados de poliol-ifn-beta. |
US7303760B2 (en) * | 1999-04-23 | 2007-12-04 | Alza Corporation | Method for treating multi-drug resistant tumors |
EP1880736A1 (en) | 1999-04-23 | 2008-01-23 | Alza Corporation | Releasable linkage and composition containing same |
EP1173221A2 (en) | 1999-04-23 | 2002-01-23 | Alza Corporation | Releasable linkage and compositions containing same |
US7238368B2 (en) | 1999-04-23 | 2007-07-03 | Alza Corporation | Releasable linkage and compositions containing same |
US7112337B2 (en) | 1999-04-23 | 2006-09-26 | Alza Corporation | Liposome composition for delivery of nucleic acid |
AU7868400A (en) | 1999-10-08 | 2001-04-23 | Alza Corporation | Neutral-cationic lipid for nucleic acid and drug delivery |
US7052686B2 (en) | 2000-09-29 | 2006-05-30 | Schering Corporation | Pegylated interleukin-10 |
US7260330B2 (en) | 2002-11-04 | 2007-08-21 | The Boeing Company | Optical communication system using correlation receiver |
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2000
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
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CN106519221A (zh) * | 2015-09-10 | 2017-03-22 | 中国科学院高能物理研究所 | 一种聚乙二醇/聚酰胺胺共聚物、其制备方法及包含该共聚物的两亲性siRNA载体 |
CN106519211A (zh) * | 2015-09-10 | 2017-03-22 | 中国科学院高能物理研究所 | 一种两亲性聚合物以及由其形成的磁性胶束纳米载体和其用途 |
CN106519221B (zh) * | 2015-09-10 | 2019-04-26 | 中国科学院高能物理研究所 | 一种聚乙二醇/聚酰胺胺共聚物、其制备方法及包含该共聚物的两亲性siRNA载体 |
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