Structure-activity relations of successful pharmacologic chaperones for rescue of naturally occurring and manufactured mutants of the gonadotropin-releasing hormone receptor
Structure-activity relations of successful pharmacologic chaperones for rescue of naturally occurring and manufactured mutants of the gonadotropin-releasing hormone receptor
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DOI：
10.1124/jpet.102.048454
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发表时间：
2003-05-01
影响因子：
3.5
通讯作者：
Conn, PM
中科院分区：
文献类型：
--
作者：
Janovick, JA;Goulet, M;Conn, PM
我们表达了野生型（WT）大鼠（r）和人（h）促性腺激素释放激素（GnRH）受体（GnRHRs）的一个测试系统，包括GnRHR的天然存在的（13种）和人工制造的（5种）“功能丧失”突变体。这些被用于评估不同的GnRH拟肽拯救有缺陷的GnRHR突变体的能力，并确定它们对野生型受体膜表达水平的影响。在人工制造的突变体中，有导致受体功能丧失的最短的大鼠GnRHR C末端截断突变体（des(325 - 327)-rGnRHR）、两个无功能的缺失突变体（des(237 - 241)-rGnRHR和des(260 - 265)-rGnRHR）、两个无功能的半胱氨酸突变体（C(229)A - rGnRHR和C(278)A - rGnRHR）；天然存在的突变体包括迄今为止报道的所有13种导致完全或部分人类低促性腺激素性性腺功能减退的全长GnRHR点突变。被评估为潜在拯救分子（“药物伴侣”）的10种拟肽来自三个不同的化学谱系（吲哚类、喹诺酮类和红霉素衍生的大环内酯类），并且最初是作为GnRH拟肽拮抗剂开发的。选择这些结构用于本研究是因为它们被预测具有穿透细胞膜并以确定的亲和力与GnRH受体相互作用的能力。所有研究的对人GnRHR的半数抑制浓度（IC50）值小于或等于2.3 nM的拟肽在拯救GnRHR突变体方面都有可测量的功效，并且在单一化学类别中，这种能力与这些IC50值相关。IC50值高达669.5 nM的红霉素衍生的大环内酯类作为拯救化合物显示出功效。拯救一种特定受体的能力是拯救其他受体能力的合理预测指标，即使跨越物种界限也是如此，尽管某些突变体不能被所测试的任何药物拯救。
We expressed a test system of wild-type (WT) rat (r) and human (h) gonadotropin-releasing hormone (GnRH) receptors (GnRHRs), including naturally occurring (13) and manufactured (five) "loss-of-function" mutants of the GnRHR. These were used to assess the ability of different GnRH peptidomimetics to rescue defective GnRHR mutants and determine their effect on the level of membrane expression of the WT receptors. Among the manufactured mutants were the shortest rGnRHR C-terminal truncation mutant that resulted in receptor loss-of-function (des(325-327)-rGnRHR), two nonfunctional deletion mutants (des(237-241)-rGnRHR and des(260-265)-rGnRHR), two nonfunctional Cys mutants (C(229)A-rGnRHR and C(278)A-rGnRHR); the naturally occurring mutants included all 13 full-length GnRHR point mutations reported to date that result in full or partial human hypogonadotropic hypogonadism. The 10 peptidomimetics assessed as potential rescue molecules ("pharmacoperones") are from three differing chemical pedigrees (indoles, quinolones, and erythromycin-derived macrolides) and were originally developed as GnRH peptidomimetic antagonists. These structures were selected for this study because of their predicted ability to permeate the cell membrane and interact with a defined affinity with the GnRH receptor. All peptidomimetics studied with an IC50 value (for hGnRHR) less than or equal to2.3 nM had measurable efficacy in rescuing GnRHR mutants, and within a single chemical class, this ability correlated to these IC50 values. Erythromycin-derived macrolides with IC50 values as high as 669.5 nM showed efficacy as rescue compounds. The ability to rescue a particular receptor was a reasonable predictor of the ability to rescue others, even across species lines, although particular mutants could not be rescued by any of the drugs tested.