Preparation and characterization of liposomal-lipophilic tumor necrosis factor.
Preparation and characterization of liposomal-lipophilic tumor necrosis factor.
复制标题
脂质体亲脂性肿瘤坏死因子的制备和表征。
作者：
T. Utsumi;M. Hung;J. Klostergaard
重组人肿瘤坏死因子 (rHuTNF) 的脂质体递送已被证明可有效降低其毒性作用并靶向富含网状内皮系统细胞的器官。然而，天然重组人 TNF 对脂质体仅表现出较差的亲和力，这可能是由于其疏水性较低。为了提高与脂质体的结合效率，我们对 TNF 进行了修饰，通过用脂肪酸侧链选择性取代其氨基来增加其疏水性。 C8 至 C18 饱和脂肪酸的 N-羟基琥珀酰亚胺酯与重组人 TNF 反应。通过天然聚丙烯酰胺凝胶电泳监测的带正电氨基的消耗来确定发生了 C8 至 C14 酸的酯的修饰；然而，较长链长的酯（C16、C18）通过酰胺键引入这些链的能力要差得多，因此没有进一步表征这些加合物。生物学测定表明，活性的保留与引入的链的数量及其长度成反比。在仅用约 1-2.5 个辛酸 (C8) 残基/三聚体修饰的 TNF 制剂中，活性最为保守（大于 50%）。发现该制剂能够高效（约 50%）与预先形成的二棕榈酰磷脂酰胆碱-小单层囊泡结合。结合的程度与引入的链的数量及其长度密切相关；对于用大约 3.5 个辛酸残基/三聚体或用大约 1.5 个肉豆蔻酸残基 (C14) 修饰的 TNF，结合效率更高 (80-90%)。然而，这些酰化 TNF 的生物活性因这种更广泛的化学修饰而进一步降低（C8 的活性低于 50%，C14 的活性低于 10%）。发现二棕榈酰磷脂酰胆碱-小单层囊泡-C8-TNF 的生物活性与非脂质体C8-TNF 相当。因此，可以高效地制备脂质体亲脂性TNF的生物活性制剂。
The liposomal delivery of recombinant human tumor necrosis factor (rHuTNF) has been shown to be effective in reducing its toxic effects and in its targeting of organs rich in cells of the reticuloendothelial system. However, native recombinant human TNF shows only poor affinity for liposomes, presumably due to its low hydrophobicity. In an attempt to increase the efficiency of association with liposomes, we have modified TNF to increase its hydrophobicity by selective substitution of its amino groups with fatty acid side chains. N-hydroxysuccinimide esters of saturated fatty acids ranging from C8 to C18 were reacted with recombinant human TNF. Modification with esters of C8 to C14 acids occurred as determined by consumption of positively charged amino groups monitored by native polyacrylamide gel electrophoresis; however, esters of longer chain lengths (C16, C18) were much less capable of introducing these chains via amide linkages, and thus these adducts were not further characterized. Biological assays revealed that retention of activity was inversely dependent both on the number of chains introduced and on their length; activity was most conserved (greater than 50%) in a TNF preparation modified with only approximately 1-2.5 caprylic acid (C8) residues/trimer. This preparation was found to bind with high efficiency (approximately 50%) to preformed dipalmitoylphosphatidylcholine-small unilamellar vesicles. The extent of binding closely paralleled both the number of chains introduced and their length; binding was even more efficient (80-90%) for TNF modified either with approximately 3.5 caprylic acid residues/trimer or with approximately 1.5 residues of myristic acid (C14). However, the biological activity of these acylated TNFs was further reduced by this more extensive chemical modification (less than 50% activity for C8 and less than 10% for C14). The biological activity of dipalmitoylphosphatidylcholine-small unilamellar vesicle-C8-TNF was found to be comparable to that of the nonliposomal C8-TNF. Thus, biologically active preparations of liposomal-lipophilic TNF can be prepared with high efficiency.
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DOI：
10.2210/pdb1tnf/pdb
发表时间：
1990-01
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
M. Eck;S. Sprang
通讯作者：
M. Eck;S. Sprang
在中和抗肿瘤坏死因子抗体存在的情况下，脂质体相关肿瘤坏死因子保留生物活性。
DOI：
--
发表时间：
1989
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Debs,RJ;Duzgunes,N;Brunette,EN;Fendly,B;Patton,J;Philip,R
通讯作者：
Philip,R
DOI：
10.1073/pnas.84.24.9265
发表时间：
1987-12-01
影响因子：
11.1
作者：
IGNARRO, LJ;BUGA, GM;CHAUDHURI, G
通讯作者：
CHAUDHURI, G
缓激肽可增加培养的豚鼠气管平滑肌细胞中的 Na( )-K( ) 泵活性。
影响因子：
7.3
作者：
Dodson,AM;Rhoden,KJ
通讯作者：
Rhoden,KJ
通过添加肉豆蔻酰化信号激活细胞原癌基因产物 p21Ras。
DOI：
10.1126/science.2648572
发表时间：
1989
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Buss,JE;Solski,PA;Schaeffer,JP;MacDonald,MJ;Der,CJ
通讯作者：
Der,CJ