Thermosensitive transient receptor potential vanilloid 2 (thermoTRPV2) is a nonselective Ca2+‐permeable cation channel broadly expressed, and is implicated in the pathology of diseases such as diabetes and pancreatitis. However, the physiological and pharmacological functions of TRPV2 channels have not been extensively investigated because of the absence of specific modulators. In this study, we report a pair of natural coumarin derivative enantiomers (−)‐murraxocin (B304‐1) and (+)‐murraxocin (B304‐2) from Murraya exotica for their selective inhibition of TRPV2 channels expressed in HEK293 cells and native TRPV2 currents in differentiated brown adipocytes. Whole‐cell patch clamp recordings confirmed the enantiomers B304‐1 and B304‐2 could selectively inhibit the agonist mediated activation of TRPV2 current with IC50 values of 22.2 ± 7.8 μM and 3.7 ± 0.7 μM, respectively. Molecular docking and site‐directed mutagenesis revealed a key residue I600 of TRPV2 critical for the binding of the enantiomers. Furthermore, B304‐1 and B304‐2 significantly reversed TRPV2 agonist‐induced inhibition of mouse brown adipocyte differentiation. Taken together, our identification of two natural coumarin enantiomers provides valuable tools and chemical leads for further elucidation of TRPV2 channel function, and pharmacological modulation of thermoTRPV2 in brown adipocytes may represent a new therapeutic strategy for treatment of energy imbalance or metabolic disorders.

中文翻译：

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鉴定用于选择性抑制 TRPV2 通道的两种天然香豆素对映体

热敏瞬时受体电位香草素 2 (thermoTRPV2) 是一种广泛表达的非选择性 Ca2+ 可渗透阳离子通道，与糖尿病和胰腺炎等疾病的病理学有关。然而，由于缺乏特异性调节剂，TRPV2 通道的生理和药理功能尚未得到广泛研究。在这项研究中，我们报告了一对来自 Murraya 奇异果的天然香豆素衍生物对映体 (-)-murraxocin (B304-1) 和 (+)-murraxocin (B304-2)，用于选择性抑制 HEK293 细胞和天然表达的 TRPV2 通道。分化的棕色脂肪细胞中的 TRPV2 电流。全细胞膜片钳记录证实对映异构体 B304-1 和 B304-2 可以选择性抑制激动剂介导的 TRPV2 电流激活，IC50 值为 22.2 ± 7.8 μM 和 3。分别为 7 ± 0.7 μM。分子对接和定点诱变揭示了 TRPV2 的一个关键残基 I600，对于对映异构体的结合至关重要。此外，B304-1 和 B304-2 显着逆转了 TRPV2 激动剂诱导的小鼠棕色脂肪细胞分化抑制。总之，我们对两种天然香豆素对映体的鉴定为进一步阐明 TRPV2 通道功能提供了有价值的工具和化学线索，棕色脂肪细胞中热 TRPV2 的药理学调节可能代表一种治疗能量失衡或代谢紊乱的新治疗策略。B304-1 和 B304-2 显着逆转了 TRPV2 激动剂对小鼠棕色脂肪细胞分化的抑制作用。总之，我们对两种天然香豆素对映体的鉴定为进一步阐明 TRPV2 通道功能提供了有价值的工具和化学线索，棕色脂肪细胞中热 TRPV2 的药理学调节可能代表一种治疗能量失衡或代谢紊乱的新治疗策略。B304-1 和 B304-2 显着逆转了 TRPV2 激动剂对小鼠棕色脂肪细胞分化的抑制作用。总之，我们对两种天然香豆素对映体的鉴定为进一步阐明 TRPV2 通道功能提供了有价值的工具和化学线索，棕色脂肪细胞中热 TRPV2 的药理学调节可能代表一种治疗能量失衡或代谢紊乱的新治疗策略。