明胶-甲氨蝶呤共轭物的分子量、药物负载量及电荷对HL-60白血病细胞生长抑制的影响。

The effect of molecular weight, drug load, and charge of gelatin-MTX conjugates on growth inhibition of HL-60 leukemia cells.

作者信息

Chen Chao-Sheng, Ofner Clyde M

机构信息

Department of Pharmaceutical Sciences, Philadelphia College of Pharmacy, University of the Sciences in Philadelphia, 600 South 43rd Street, Philadelphia, Pennsylvania 19104, USA.

出版信息

Pharm Res. 2009 Feb;26(2):338-45. doi: 10.1007/s11095-008-9746-5. Epub 2008 Oct 31.

DOI:10.1007/s11095-008-9746-5

PMID:18975058

Abstract

PURPOSE

Gelatin-methotrexate conjugates (G-MTX) with known molecular weight (MW), drug load, and charge were prepared and evaluated for growth inhibition on leukemia cells.

METHODS

Gelatin (34 to 171 kDa) was reacted with a carbodiimide to prepare G-MTX with high (G-MTX-H) and low (G-MTX-L) drug loads. Cationic conjugates were prepared by ethylenediamine modification. MTX:gelatin molar ratios were determined spectrophotometrically. Isoelectric focusing electrophoresis (IEF) and turbidity were used to measure isoelectric points (IEP). Growth inhibition profiles and IC50 values were determined on HL-60 cells using a modified MTT assay.

RESULTS

IC50 values of anionic G-MTX-L (drug loads 0.5:1 to 2.2:1) increased linearly from 46 to 180 nM with MW. But, IC50 values for anionic G-MTX-H (drug loads 7.4:1 to 25:1) showed little, if any, MW dependence and were about two times higher. IC50 values for cationic G-MTX-L ranged from 770 to 2,900 nM and the relationship with MW was non-linear.

CONCLUSIONS

The growth inhibition ranking was MTX>anionic G-MTX-L>anionic G-MTX-H>cationic G-MTX-L. High drug load may hinder lysosomal enzyme degradation and drug release and contribute to suppression of the MW effect observed with G-MTX-L. A mechanism change is suggested as the cationic conjugates increase to the highest MW.

摘要

目的

制备具有已知分子量（MW）、载药量和电荷的明胶 - 甲氨蝶呤缀合物（G - MTX），并评估其对白血病细胞的生长抑制作用。

方法

将明胶（34至171 kDa）与碳二亚胺反应，制备高载药量（G - MTX - H）和低载药量（G - MTX - L）的G - MTX。通过乙二胺修饰制备阳离子缀合物。用分光光度法测定MTX与明胶的摩尔比。采用等电聚焦电泳（IEF）和浊度法测量等电点（IEP）。使用改良的MTT法测定HL - 60细胞的生长抑制曲线和IC50值。

结果

阴离子型G - MTX - L（载药量0.5:1至2.2:1）的IC50值随分子量从46 nM线性增加至180 nM。但是，阴离子型G - MTX - H（载药量7.4:1至25:1）的IC50值几乎与分子量无关，即使有关系也很小，且约高两倍。阳离子型G - MTX - L的IC50值范围为770至2900 nM，与分子量的关系是非线性的。