比马前列素、曲伏前列素、乌诺前列酮及其他FP前列腺素受体激动剂类似物诱导的人小梁网细胞反应。

Human trabecular meshwork cell responses induced by bimatoprost, travoprost, unoprostone, and other FP prostaglandin receptor agonist analogues.

作者信息

Sharif Najam A, Kelly Curtis R, Crider Julie Y

机构信息

Molecular Pharmacology Unit, Glaucoma Research, Alcon Research, Ltd., Fort Worth, Texas, USA.

出版信息

Invest Ophthalmol Vis Sci. 2003 Feb;44(2):715-21. doi: 10.1167/iovs.02-0323.

DOI:10.1167/iovs.02-0323

PMID:12556403

Abstract

PURPOSE

To determine the functional agonist potencies of the intraocular pressure (IOP)-lowering prostaglandin F (FP)-class prostaglandin (PG) analogues (e.g., travoprost, latanoprost, bimatoprost, and unoprostone isopropyl ester) in human trabecular meshwork (h-TM) cells, by using phosphoinositide (PI) turnover and intracellular Ca(2+) (Ca(2+)) mobilization, and to confirm the FP nature of these receptors by using an FP receptor antagonist, 11beta-fluoro-15-epi-15-indanyl-PGF(2alpha) (AL-8810).

METHODS

FP-receptor-mediated PI turnover and Ca(2+) mobilization were measured in h-TM cells by determining the accumulation of [(3)H]-inositol phosphates ([(3)H]-IPs) by anion-exchange chromatography and real-time fluorescence imaging, respectively.

RESULTS

Various PG analogues concentration-dependently stimulated production of [(3)H]-IPs in h-TM cells with the following agonist potencies (median effective concentration; EC(50)): travoprost acid (EC(50) = 2.4 nM) > cloprostenol (EC(50) = 4.5 nM) > (+/-)-fluprostenol (EC(50) = 10.8 nM) > latanoprost acid (EC(50) = 34.7 nM) > bimatoprost acid (EC(50) = 112 nM) > PGF(2alpha) (EC(50) = 120 nM) >> unoprostone (UF-021; EC(50) = 3280 nM) > S-1033 (EC(50) = 4570 nM; all n = 3-9). Prodrug derivatives of these compounds exhibited the following potencies: travoprost (isopropyl ester; EC(50) = 89.1 nM) > latanoprost (isopropyl ester; EC(50) = 778 nM) > bimatoprost (amide; EC(50) = 1410-6940 nM). Travoprost acid, PGF(2alpha,) unoprostone, and S-1033 were tested in addition for Ca(2+) mobilization and found to have rapid and dose-dependent effects. The FP receptor-selective antagonist AL-8810 antagonized the (+/-)-fluprostenol-induced PI turnover in these cells (K(i) = 2.56 +/- 0.62 micro M) as well as that induced by bimatoprost and acids of latanoprost and travoprost. The agonist and antagonist potencies of the PG analogues from the PI turnover assays in h-TM cells correlated well with PI turnover data obtained from the cloned human ciliary body FP receptor (r = 0.92; P < 0.0001).

CONCLUSIONS

The pharmacology of the h-TM cell FP-receptor-mediated PI turnover and Ca(2+) mobilization was defined using numerous synthetic (FP-selective) PG agonist analogues and an FP receptor antagonist, AL-8810. Bimatoprost, travoprost, latanoprost, unoprostone isopropyl ester, and their respective free acids were shown to be FP agonists in the h-TM cells.

摘要

目的

通过磷酸肌醇（PI）周转和细胞内钙离子（[Ca²⁺]i）动员，测定降低眼压的前列腺素F（FP）类前列腺素（PG）类似物（如曲伏前列素、拉坦前列素、比马前列素和异丙酸乌诺前列酮）在人小梁网（h-TM）细胞中的功能激动剂效力，并使用FP受体拮抗剂11β-氟-15-表-15-茚满基-PGF₂α（AL-8810）确认这些受体的FP性质。

方法

分别通过阴离子交换色谱法测定[³H]-肌醇磷酸（[³H]-IPs）的积累和实时荧光成像，在h-TM细胞中测量FP受体介导的PI周转和[Ca²⁺]i动员。

结果

各种PG类似物在h-TM细胞中浓度依赖性地刺激[³H]-IPs的产生，其激动剂效力如下（半数有效浓度；EC₅₀）：曲伏前列酸（EC₅₀ = 2.4 nM）>氯前列醇（EC₅₀ = 4.5 nM）>（±）-氟前列醇（EC₅₀ = 10.8 nM）>拉坦前列酸（EC₅₀ = 34.7 nM）>比马前列酸（EC₅₀ = 112 nM）>PGF₂α（EC₅₀ = 120 nM）>>异丙酸乌诺前列酮（UF-021；EC₅₀ = 3280 nM）>S-1033（EC₅₀ = 4570 nM；所有n = 3 - 9）。这些化合物的前药衍生物表现出以下效力：曲伏前列素（异丙酯；EC₅₀ = 89.1 nM）>拉坦前列素（异丙酯；EC₅₀ = 778 nM）>比马前列素（酰胺；EC₅₀ = 1410 - 6940 nM）。此外，还测试了曲伏前列酸、PGF₂α、异丙酸乌诺前列酮和S-1033的[Ca²⁺]i动员情况，发现它们具有快速且剂量依赖性的作用。FP受体选择性拮抗剂AL-8810拮抗了这些细胞中（±）-氟前列醇诱导的PI周转（K i = 2.56 ± 0.62 μM）以及比马前列素和拉坦前列素及曲伏前列素酸诱导的PI周转。h-TM细胞中PI周转试验得到的PG类似物的激动剂和拮抗剂效力与从克隆的人睫状体FP受体获得的PI周转数据相关性良好（r = 0.92；P < 0.0001）。