设计对人催产素受体具有比阿托西班显著更高亲和力和选择性的肽类催产素拮抗剂。

Design of peptide oxytocin antagonists with strikingly higher affinities and selectivities for the human oxytocin receptor than atosiban.

作者信息

Manning Maurice, Cheng Ling Ling, Stoev Stoytcho, Wo Nga Ching, Chan W Y, Szeto Hazel H, Durroux Thierry, Mouillac Bernard, Barberis Claude

机构信息

Department of Biochemistry and Cancer Biology, Medical College of Ohio, Toledo, OH 43614, USA.

出版信息

J Pept Sci. 2005 Oct;11(10):593-608. doi: 10.1002/psc.667.

DOI:10.1002/psc.667

PMID:15880385

Abstract

The peptide oxytocin (OT) antagonist atosiban, approved for tocolytic use in Europe (under the tradename Tractocile), represents an important new therapeutic advance for the treatment of premature labor. This paper presents some new peptide OT antagonists which offer promise as superior tocolytics. The solid phase synthesis is reported of four pairs of L and D-2-naphthylalanine (L/D-2Nal) position-2 modified analogs of the following four oxytocin (OT) antagonists: des-9-glycinamide [1-(beta-mercapto-beta,beta-pentamethylene propionic acid), 2-O-methyltyrosine, 4-threonine]ornithine-vasotocin (desGly-NH(2),d(CH(2))(5)[Tyr(Me)(2),Thr(4)]OVT) (A); the Tyr-NH(2) (9) analog of (A), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Tyr-NH(2) (9)]OVT (B); the Eda(9) analog of (A), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Eda(9)]OVT (C); and the retro COCH(2)Ph(4-0H)(10) modified analog of (C), d(CH(2))(5)[Tyr(Me)(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-0H)(10)]OVT (D). The eight new analogs of A-D are (1) desGly-NH(2),d(CH(2))(5)[D-2Nal(2),Thr(4)]OVT, (2) desGly-NH(2),d(CH(2))(5)[2-Nal(2),Thr(4)]OVT, (3) d(CH(2))(5)[D-2Nal(2),Thr(4),Tyr-NH(2) (9)]OVT, (4) d(CH(2))(5)[2Nal(2),Thr(4),Tyr-NH(2) (9)]OVT, (5) d(CH(2))(5)[D-2Nal(2),Thr(4),Eda(9)]OVT, (6) d(CH(2))(5)[2Nal(2),Thr(4),Eda(9)]OVT, (7) d(CH(2))(5)[D-2Nal(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-0H)(10)]OVT, (8) d(CH(2))(5)[2Nal(2),Thr(4),Eda(9)<-- COCH(2)Ph(4-OH)(10)]OVT. Peptides 1-8 were evaluated for agonistic and antagonistic activities in in vitro and in vivo rat bioassays, in rat OT receptor (rOTR) binding assays and in human OT receptor (hOTR) and human vasopressin (VP) vasopressor (V(1a)) receptor (hV(1a)R) binding assays. Also reported are the hOTR and hV(1a)R affinity data for atosiban and for B. None of the eight peptides exhibit oxytocic or vasopressor agonism. Peptides 1-8 exhibit weak antidiuretic agonism (activities in the range 0.014-0.21 U/mg). Peptides 1-6 exhibit potent in vitro (no Mg(2+)) OT antagonism (anti-OT pA(2) values range from 7.63 to 8.08). Peptides 7 and 8 are weaker OT antagonists. Peptides 1-6 are all OT antagonists in vivo (estimated in vivo anti-OT pA(2) values in the range 6.94-7.23). Peptides 1-8 exhibit vasopressor antagonism, anti-V(1a) pA(2) values in the range 5.1-7.65. Peptides 1-8 exhibit high affinities for the rOTR (K(i) values = 0.3-7.8 nM). Peptides 1-4 and B exhibit surprisingly very high affinities for the hOTR; their K(i) values are 0.17, 0.29, 0.07, 0.14 and 0.59 nM, respectively. Peptides 1-4 and B exhibit respectively 449, 263, 1091, 546 and 129 times greater affinity for the hOTR than atosiban (K(i) = 76.4 nM). Peptides 1-4 exhibit high affinities for the hV(1a)R (K(i)s = 1.1 nM, 1.3 nM, 0.19 nM and 0.54 nM, all higher than the hV1(a)R affinities exhibited by atosiban (K(i) = 5.1 nM) and by B (K(i) = 5.26 nM). Because of their strikingly higher affinities for the hOTR than atosiban, peptides 1-4 and B exhibit gains in anti hOT/anti hV(1a) receptor selectivity compared with atosiban of 93, 64, 39, 56 and 127, respectively. These OT antagonists are thus promising candidates for development as potential new tocolytic agents.

摘要

肽类催产素（OT）拮抗剂阿托西班在欧洲被批准用于安胎治疗（商品名为Tractocile），它代表了早产治疗方面一项重要的新治疗进展。本文介绍了一些有望成为更优安胎药的新型肽类OT拮抗剂。报道了以下四种催产素（OT）拮抗剂的四对L型和D型2-萘丙氨酸（L/D-2Nal）在2位修饰的类似物的固相合成：去-9-甘氨酰胺[1-(β-巯基-β,β-戊亚甲基丙酸)，2-O-甲基酪氨酸，4-苏氨酸]鸟氨酸加压素（desGly-NH(2)，d(CH(2))(5)[Tyr(Me)(2)，Thr(4)]OVT）（A）；（A）的Tyr-NH(2)（9）类似物，d(CH(2))(5)[Tyr(Me)(2)，Thr(4)，Tyr-NH(2)（9）]OVT（B）；（A）的Eda(9)类似物，d(CH(2))(5)[Tyr(Me)(2)，Thr(4)，Eda(9)]OVT（C）；以及（C）的反向COCH(2)Ph(4-OH)(10)修饰类似物，d(CH(2))(5)[Tyr(Me)(2)，Thr(4)，Eda(9)<--COCH(2)Ph(4-OH)(l0)]OVT（D）。A-D的八种新类似物分别为：（1）desGly-NH(2)，d(CH(2))(5)[D-2Nal(2)，Thr(4)]OVT，（2）desGly-NH(2)，d(CH(2))(5)[2-Nal(2)，Thr(4)]OVT，（3）d(CH(2))(5)[D-2Nal(2)，Thr(4)，Tyr-NH(2)（9）]OVT，（4）d(CH(2))(5)[2Nal(2)，Thr(4)，Tyr-NH(2)（9）]OVT，（5）d(CH(2))(5)[D-2Nal(2)，Thr(4)，Eda(9)]OVT，（6）d(CH(2))(5)[2Nal(2)，Thr(4)，Eda(9)]OVT，（7）d(CH(2))(5)[D-2Nal(2)，Thr(4)，Eda(9)<--COCH(2)Ph(4-OH)(10)]OVT，（8）d(CH(2))(5)[2Nal(2)，Thr(4)，Eda(9)<--COCH(2)Ph(4-OH)(10)]OVT。在体外和体内大鼠生物测定、大鼠OT受体（rOTR）结合测定以及人OT受体（hOTR）和人血管加压素（VP）升压（V(1a)）受体（hV(1a)R）结合测定中对肽1-8的激动和拮抗活性进行了评估。还报道了阿托西班和B的hOTR及hV(1a)R亲和力数据。八种肽均未表现出催产或升压激动作用。肽1-8表现出较弱的抗利尿激动作用（活性范围为0.014 - 0.21 U/mg）。肽1-6在体外（无Mg(2+)）表现出强效的OT拮抗作用（抗OT pA(2)值范围为7.63至8.08）。肽7和8是较弱的OT拮抗剂。肽1-6在体内均为OT拮抗剂（体内估计的抗OT pA(2)值范围为6.94 - 7.23）。肽1-8表现出升压拮抗作用，抗V(1a) pA(2)值范围为5.1 - 7.65。肽1-8对rOTR具有高亲和力（K(i)值 = 0.3 - 7.8 nM）。肽1-4和B对hOTR表现出惊人的高亲和力；它们的K(i)值分别为0.17、0.29、0.07、0.14和0.59 nM。肽1-4和B对hOTR的亲和力分别比阿托西班（K(i) = 76.4 nM）高449、263、1091、546和129倍。肽1-4对hV(1a)R具有高亲和力（K(i)分别为1.1 nM、1.3 nM、0.19 nM和0.54 nM，均高于阿托西班（K(i) = 5.1 nM）和B（K(i) = 5.26 nM）所表现出的hV1(a)R亲和力）。由于肽1-4和B对hOTR的亲和力显著高于阿托西班，它们与阿托西班相比，抗hOT/抗hV(1a)受体选择性分别提高了93、64、39、56和127倍。因此，这些OT拮抗剂有望成为潜在的新型安胎药开发候选药物。