新型抗HIV药物——科萨烷及其同系物在人肠上皮（Caco-2）细胞单层中的转运增强。

Enhanced transport of a novel anti-HIV agent--cosalane and its congeners across human intestinal epithelial (Caco-2) cell monolayers.

作者信息

Udata Chandrasekhar, Patel Jignesh, Pal Dhananjay, Hejchman Elzbieta, Cushman Mark, Mitra Ashim K

机构信息

Drug Metabolism, Wyeth Research, Pearl River, NY, USA.

出版信息

Int J Pharm. 2003 Jan 2;250(1):157-68. doi: 10.1016/s0378-5173(02)00523-9.

DOI:10.1016/s0378-5173(02)00523-9

PMID:12480282

Abstract

PURPOSE

Cosalane is a potent inhibitor of HIV replication with activity against a broad range of viral targets. However, oral bioavailability of this highly lipophilic compound is extremely poor (<1%). The purpose of this study is to screen a variety of permeation enhancers (cyclodextrin derivatives, cremophor EL, bile salts and mixed micelles) for their ability to enhance the transport of cosalane and its analogs/prodrugs across Caco-2 cell monolayers.

METHODS

Cosalane and its different analogs/prodrugs were synthesized and their physicochemical properties were determined. Caco-2 cells were cultured at a density of 66,000 cells/cm(2) either on collagen coated clear polyester membranes or Transwell inserts. Side-bi-side diffusion cells and Transwell inserts were employed to study for the transport of cosalane and its analogs/prodrugs with various permeation enhancers across Caco-2 cell monolayers.

RESULTS

Permeabilities of EH-3-39, EH-3-55 and EH-3-57 significantly improved compared to that of cosalane in the presence of bile salt, sodium desoxycholate. Among the various cyclodextrins studied, hydroxypropyl beta cyclodextrin (HP-beta-CD) and dimethyl beta cyclodextrin (DM-beta-CD) exhibited 22.3-fold and 19-fold permeability enhancement of cosalane respectively across Caco-2 cell monolayers. Sodium desoxycholate (10 mM) also showed a remarkable (105-fold) enhancement on the permeability of cosalane (P(app) 11.72+/-3.31 x 10(-6) cm/s) without causing any measurable cellular damage. Cremophor EL resulted in higher transport of 14C mannitol. The mechanism of enhancement effect can be mainly attributed to the alteration of membrane fluidity by cyclodextrin and opening of tight junctions by cremophor EL.

CONCLUSIONS

Among the enhancers tested, 10 mM sodium desoxycholate and HP-beta-CD appear to be viable candidates for further development of an oral formulation of cosalane and its congeners.

摘要

目的

科萨烷是一种强效的HIV复制抑制剂，对多种病毒靶点具有活性。然而，这种高度亲脂性化合物的口服生物利用度极低（<1%）。本研究的目的是筛选多种渗透促进剂（环糊精衍生物、聚氧乙烯蓖麻油EL、胆盐和混合胶束），以评估它们增强科萨烷及其类似物/前体药物跨Caco-2细胞单层转运的能力。

方法

合成了科萨烷及其不同的类似物/前体药物，并测定了它们的理化性质。将Caco-2细胞以66,000个细胞/cm²的密度接种在胶原包被的透明聚酯膜或Transwell小室上。采用并排扩散池和Transwell小室研究科萨烷及其类似物/前体药物与各种渗透促进剂跨Caco-2细胞单层的转运。

结果

在胆盐脱氧胆酸钠存在的情况下，EH-3-39、EH-3-55和EH-3-57的渗透率与科萨烷相比显著提高。在所研究的各种环糊精中，羟丙基-β-环糊精（HP-β-CD）和二甲基-β-环糊精（DM-β-CD）使科萨烷跨Caco-2细胞单层的渗透率分别提高了22.3倍和19倍。脱氧胆酸钠（10 mM）也使科萨烷的渗透率显著提高（105倍）（P(app) 11.72±3.31×10⁻⁶ cm/s），且未造成任何可测量的细胞损伤。聚氧乙烯蓖麻油EL导致14C甘露醇的转运增加。增强作用的机制主要可归因于环糊精改变膜流动性以及聚氧乙烯蓖麻油EL打开紧密连接。