Synthesis and in vitro characterization of novel dextran-methylprednisolone conjugates with peptide linkers: effects of linker length on hydrolytic and enzymatic release of methylprednisolone and its peptidyl intermediates.
Synthesis and in vitro characterization of novel dextran-methylprednisolone conjugates with peptide linkers: effects of linker length on hydrolytic and enzymatic release of methylprednisolone and its peptidyl intermediates.
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DOI：
10.1002/jps.21161
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发表时间：
2008-07
影响因子：
3.8
通讯作者：
Mehvar, Reza
中科院分区：
文献类型：
--
作者：
Penugonda, Suman;Kumar, Anil;Agarwal, Hitesh K.;Parang, Keykavous;Mehvar, Reza
关键词：
为了控制溶酶体中甲基强的松龙 (MP) 的释放速率，合成并表征了带有肽接头的新型葡聚糖-MP 缀合物。使用具有 1-5 个甘氨酸残基的接头将甲基强的松龙琥珀酸酯 (MPS) 连接至 25 kDa 的葡聚糖。使用尺寸排阻和/或新开发的反相 HPLC 方法测定缀合物在 pH 4.0 和 7.4 缓冲液、血液、肝溶酶体和各种溶酶体蛋白酶中的释放特性，该方法能够同时定量 MP、MPS 和所有五种可能的 MPS-肽基中间体。我们合成的缀合物纯度≥ 90%，MP 取代度为 6.9–9.5% (w/w)。该缀合物在 pH 4.0 下稳定，但在 pH 7.4 缓冲液和大鼠血液中释放 MP 和完整的 MPS-肽基中间体，较长连接体的降解速率更快。大鼠溶酶体部分将缀合物降解为 MP 和所有可能的中间体，降解速度也与肽的长度成正比。虽然半胱氨酸肽酶（木瓜蛋白酶或组织蛋白酶 B）对缀合物的降解相对较大，但在天冬氨酸（组织蛋白酶 D）或丝氨酸（胰蛋白酶）蛋白酶存在的情况下没有观察到降解，这些蛋白酶不会用 Gly 裂解肽键。这些新开发的 MP 葡聚糖缀合物有望在溶酶体中控制 MP 递送。
To control the rate of release of methylprednisolone (MP) in lysosomes, new dextran-MP conjugates with peptide linkers were synthesized and characterized. Methylprednisolone succinate (MPS) was attached to dextran 25 kDa using linkers with 1–5 Gly residues. The release characteristics of the conjugates in pH 4.0 and 7.4 buffers, blood, liver lysosomes, and various lysosomal proteinases were determined using a size-exclusion and/or a newly-developed reversed-phase HPLC method capable of simultaneous quantitation of MP, MPS, and all five possible MPS-peptidyl intermediates. We synthesized conjugates with ≥ 90% purity and 6.9–9.5% (w/w) degree of MP substitution. The conjugates were stable at pH 4.0, but released MP and intact MPS-peptidyl intermediates in the pH 7.4 buffer and rat blood, with faster degradation rates for longer linkers. Rat lysosomal fractions degraded the conjugates to MP and all the possible intermediates also at a rate directly proportional to the length of the peptide. Whereas the degradation of the conjugates by cysteine peptidases (papain or cathepsin B) was relatively substantial, no degradation was observed in the presence of aspartic (cathepsin D) or serine (trypsin) proteinases, which do not cleave peptide bonds with Gly. These newly-developed dextran conjugates of MP show promise for controlled delivery of MP in lysosomes.
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影响因子：
16.1
作者：
LARSEN C
通讯作者：
LARSEN C
影响因子：
3.7
作者：
Chimalakonda, AP;Mehvar, R
通讯作者：
Mehvar, R
影响因子：
6.2
作者：
Chimalakonda, AP;Montgomery, DL;Mehvar, R
通讯作者：
Mehvar, R
影响因子：
2.8
作者：
Mehvar, Reza
通讯作者：
Mehvar, Reza
影响因子：
5.8
作者：
Rensberger, KL;Hoganson, DA;Mehvar, R
通讯作者：
Mehvar, R