二氢丹参酮 I 靶向刺突蛋白 N 端结构域以抑制 SARS-CoV-2 进入及刺突蛋白诱导的炎症反应。

Dihydrotanshinone I targets the spike N-terminal domain to inhibit SARS-CoV-2 entry and spike-induced inflammation.

作者信息

Yang Chan, Lu Kunyu, Wang Jinshen, Su Hao, Cheng Luping, Nong Chunmei, Pan Xiaoyan, Li Haobin, Peng Zheng, Chen Yinghua, Wu Yan, Ma Qinhai, Hu Guodong, Liu Shuwen

机构信息

Department of Respiratory and Critical Care Medicine, Institute of Respiratory and Critical Care Medicine, Dongguan People's Hospital Biobank, the Tenth Affiliated Hospital of Southern Medical University, Southern Medical University, Guangdong 523059, PR China; NMPA Key Laboratory for Research and Evaluation of Drug Metabolism & Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, PR China.

NMPA Key Laboratory for Research and Evaluation of Drug Metabolism & Guangdong Provincial Key Laboratory of New Drug Screening, School of Pharmaceutical Sciences, Southern Medical University, Guangzhou 510515, PR China.

出版信息

Phytomedicine. 2025 Sep;145:156989. doi: 10.1016/j.phymed.2025.156989. Epub 2025 Jun 17.

DOI:10.1016/j.phymed.2025.156989

PMID:40570554

Abstract

BACKGROUND

Despite progress made in addressing the COVID-19 pandemic, numerous challenges persist, including breakthrough infections (BTI) and reinfections with SARS-CoV-2. There is an urgent need for broad-spectrum antiviral agents with multifaceted functionalities. Dihydrotanshinone I (DHT), a lipophilic monomer component of Salvia miltiorrhiza, exhibits diverse pharmacological activities, including antioxidant, anti-inflammatory, antibacterial and immunomodulatory effects. The efficacy and mechanisms underlying its "multi-target" advantages in treating COVID-19 warrant further investigation.

PURPOSE

To investigate the inhibitory activity and mechanisms of DHT against pan-SARS-CoV-2 invasion.

METHODS

We examined the activity of DHT against SARS-CoV-2 Variants of Concern (VOCs) using the viral replicon system, pseudo virus-based entry assays, and a human ACE2 transgenic mouse model. The enzymatic activity and time-of-drug-addition assays were conducted to elucidate the viral life cycle targeted by DHT. Drug resistance induction assay, next-generation sequencing (NGS) analysis and molecular docking were employed to confirm DHT's target sites. Further ELISA, RT-qPCR, and western blot assays revealed the mechanism and anti-inflammatory activity of DHT.

RESULTS

As a potent antiviral agent, DHT showed activity against SARS-CoV-2 VOCs with EC values ranging from 0.3-4.0 μM. DHT binds to two conserved sites on the spike N-terminal domain (NTD), inhibiting viral entry for both original and variant strains. Furthermore, DHT modulates the interaction between the spike and Toll-like receptor 4 (TLR4), stimulating nuclear factor-erythroid 2-related factor 2 (NRF2)-dependent immune response to attenuate cytokine production. Moreover, multiple administrations of DHT provide therapeutic benefits in mouse model of COVID-19.

CONCLUSIONS

The dual functionality of DHT is accomplished through binding to the spike NTD and inhibiting spike-activated inflammation, making it effective in preventing SARS-CoV-2 invasion. Our findings offer a conceptual framework for the rational design of prospective multitargeted inhibitors against SARS-CoV-2 spike NTD.

摘要

背景

尽管在应对新冠疫情方面取得了进展，但仍存在诸多挑战，包括突破性感染（BTI）和新冠病毒再感染。迫切需要具有多方面功能的广谱抗病毒药物。二氢丹参酮I（DHT）是丹参的一种亲脂性单体成分，具有多种药理活性，包括抗氧化、抗炎、抗菌和免疫调节作用。其在治疗新冠病毒方面“多靶点”优势的疗效和机制值得进一步研究。

目的

研究DHT对泛新冠病毒入侵的抑制活性及机制。

方法

我们使用病毒复制子系统、基于假病毒的进入试验和人血管紧张素转换酶2（ACE2）转基因小鼠模型，检测了DHT对新冠病毒变异株（VOCs）的活性。进行酶活性和药物添加时间试验以阐明DHT靶向的病毒生命周期。采用耐药诱导试验、二代测序（NGS）分析和分子对接来确认DHT的靶点。进一步的酶联免疫吸附测定（ELISA）、逆转录定量聚合酶链反应（RT-qPCR）和蛋白质印迹分析揭示了DHT的作用机制和抗炎活性。

结果

作为一种有效的抗病毒药物，DHT对新冠病毒变异株表现出活性，半数有效浓度（EC）值范围为0.3 - 4.0微摩尔。DHT与刺突蛋白N端结构域（NTD）上的两个保守位点结合，抑制原始毒株和变异毒株的病毒进入。此外，DHT调节刺突蛋白与Toll样受体4（TLR4）之间的相互作用，刺激核因子红细胞2相关因子2（NRF2）依赖性免疫反应，以减少细胞因子的产生。此外，多次给予DHT对新冠病毒小鼠模型具有治疗益处。