Identification and Characterization of Novel Small-Molecule Protease-Activated Receptor 2 Agonists
Identification and Characterization of Novel Small-Molecule Protease-Activated Receptor 2 Agonists
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DOI：
10.1124/jpet.108.142570
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发表时间：
2008-12-01
影响因子：
3.5
通讯作者：
Burstein, Ethan S.
中科院分区：
文献类型：
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作者：
Gardell, Luis R.;Ma, Jian-Nong;Burstein, Ethan S.
我们报告了第一个小分子蛋白酶激活受体（PAR）2激动剂，AC-55541 [N-[[1-（3-溴-苯基）-乙-（E）-亚基-肼基羰基]-（4-氧代-3，4-二氢-酞嗪-1-基）-甲基]-苯甲酰胺]和AC-264613 [2-氧代-4-苯基吡咯烷-3-羧酸[1-（3-溴-苯基）-（E/Z）亚乙基]-酰肼]，各自代表不同的化学系列。在细胞增殖试验、磷脂酰肌醇水解试验和Ca 2+动员试验中，AC-55541和AC-264613均激活PAR 2信号传导，AC-55541的效价范围为200 - 1000 nM，AC-264613的效价范围为30 - 100 nM。相比之下，PAR 2活化肽2-呋喃甲酰基-LIGRL 0-NH 2具有相似的效力，而SLIGRL-NH 2的效力低30至300倍。AC-55541和AC-264613对任何其他PAR受体亚型都没有活性，对参与伤害感受的超过30种其他分子靶点也没有任何显著的亲和力。AC-55541和AC-264613在大鼠腹腔内给药时吸收良好，均达到微摩尔血浆峰浓度。AC-55541和AC-264613均对肝微粒体代谢稳定，并在大鼠中保持持续暴露，消除半衰期分别为6.1和2.5 h。AC-55541或AC-264613爪内给药引起了强烈和持续的热痛觉过敏和水肿。同时给予速激肽1（神经激肽1）受体拮抗剂或瞬时受体电位香草酸（TRPV）1拮抗剂完全阻断了这些作用。AC-55541或AC-264613全身给药产生的痛觉过敏程度与化合物局部给药时观察到的相似。这些化合物代表了新的小分子PAR 2激动剂，其将用于探测PAR 2受体的生理功能。
We report the first small-molecule protease-activated receptor (PAR) 2 agonists, AC-55541 [N-[[1-(3-bromo-phenyl)-eth-(E)-ylidene-hydrazinocarbonyl]-(4-oxo-3,4-dihydro-phthalazin-1-yl)-methyl]-benzamide] and AC-264613 [2-oxo-4-phenylpyrrolidine-3-carboxylic acid [1-(3-bromo-phenyl)-(E/Z)ethylidene]-hydrazide], each representing a distinct chemical series. AC-55541 and AC-264613 each activated PAR2 signaling in cellular proliferation assays, phosphatidylinositol hydrolysis assays, and Ca2+ mobilization assays, with potencies ranging from 200 to 1000 nM for AC-55541 and 30 to 100 nM for AC-264613. In comparison, the PAR2-activating peptide 2-furoyl-LIGRLO-NH2 had similar potency, whereas SLIGRL-NH2 was 30 to 300 times less potent. Neither AC-55541 nor AC-264613 had activity at any of the other PAR receptor subtypes, nor did they have any significant affinity for over 30 other molecular targets involved in nociception.Visualization of EYFP-tagged PAR2 receptors showed that each compound stimulated internalization of PAR2 receptors. AC-55541 and AC-264613 were well absorbed when administered intraperitoneally to rats, each reaching micromolar peak plasma concentrations. AC-55541 and AC-264613 were each stable to metabolism by liver microsomes and maintained sustained exposure in rats, with elimination half-lives of 6.1 and 2.5 h, respectively. Intrapaw administration of AC-55541 or AC-264613 elicited robust and persistent thermal hyperalgesia and edema. Coadministration of either a tachykinin 1 (neurokinin 1) receptor antagonist or a transient receptor potential vanilloid (TRPV) 1 antagonist completely blocked these effects. Systemic administration of either AC-55541 or AC-264613 produced a similar degree of hyperalgesia as was observed when the compounds were administered locally. These compounds represent novel small-molecule PAR2 agonists that will be useful in probing the physiological functions of PAR2 receptors.