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壳聚糖辅助固定化木瓜蛋白酶于磁性纳米粒子、其表征及其靶向递药。

Chitosan-assisted immobilization of serratiopeptidase on magnetic nanoparticles, characterization and its target delivery.

机构信息

Department of Biotechnology, Dr. B.R. Ambedkar National Institute of Technology , Jalandhar, Punjab , India .

出版信息

J Drug Target. 2014 Feb;22(2):123-37. doi: 10.3109/1061186X.2013.844157. Epub 2013 Oct 23.

Abstract

BACKGROUND

Magnetic nanoparticles (MNPs) gained attentions as universal carrier for drug delivery and for enzyme immobilization.

PURPOSE

Target delivery of serratiopeptidase for treatment using this enzyme and applications in drug delivery.

METHOD

Serratiopeptidase was immobilized on chitosan amino-functionalized MNPs by covalent bonding through glutaraldehyde. Targeting of MNPs with immobilized enzyme (EMNPs) was carried out in vitro in modified diffusion cell and in vivo in rats.

RESULTS AND DISCUSSION

MNPs and EMNPs were 15.43 ± 5.22 and 18.43 ± 3.23 nm (transmission electron microscopy), crystallite size 16.89 and 21.05 nm (X-ray diffraction) and saturation magnetization 62 and 35.2 emug(-1), respectively. Maximum protein and enzyme loading on EMNPs were 264 mg g(-1) and 325 U g(-1), respectively. In the molecular level, maximum 52 enzyme molecules could bind to each particle showing residual activity 68%, little effect on KM and Vmax, good storage stability. Magnetic targeting of EMNPs increased the delivery (permeation) of drug through membrane in vitro and the enhanced anti-inflammatory effect on carrageenan-induced paw oedema in rats in vivo at much lower doses of enzyme than the doses required for treatment with free enzyme.

CONCLUSIONS

The enzymatic preparation of MNPs showed enhanced effects (permeation enhancement and anti-inflammatory activity) at lower concentration with magnetic targeting.

摘要

背景

磁性纳米颗粒(MNPs)作为药物输送和酶固定化的通用载体引起了人们的关注。

目的

使用这种酶进行糜蛋白酶的靶向递送治疗,并将其应用于药物输送。

方法

通过戊二醛共价键将糜蛋白酶固定在壳聚糖氨基功能化的 MNPs 上。通过体外改良扩散池和体内大鼠进行 MNPs 与固定化酶(EMNPs)的靶向。

结果与讨论

MNPs 和 EMNPs 的粒径分别为 15.43±5.22nm 和 18.43±3.23nm(透射电子显微镜),结晶度分别为 16.89nm 和 21.05nm(X 射线衍射),饱和磁化强度分别为 62 和 35.2 emu/g。EMNPs 的最大蛋白和酶载量分别为 264mg/g 和 325U/g。在分子水平上,每个颗粒最多可结合 52 个酶分子,表现出 68%的残余活性,对 KM 和 Vmax 影响较小,具有良好的储存稳定性。EMNPs 的磁靶向增加了药物在体外通过膜的传递,以及在体内角叉菜胶诱导的足肿胀中增强抗炎作用,所需酶的剂量比用游离酶治疗所需的剂量低得多。

结论

MNPs 的酶制剂在较低浓度下通过磁靶向显示出增强的效果(渗透增强和抗炎活性)。

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