JinHuangJieDu (JHJD) formula attenuates SARS-CoV-2 infection by interrupting RBD-ACE2 binding and HIF-1α-dependent inflammation

Background Since the emergence and spread of coronavirus disease 2019 (COVID-19), Traditional Chinese Medicine (TCM) and its formulations have been used extensively in China for preventing and treating COVID-19 with demonstrated clinical efficacy. This study aimed to evaluate both the therapeutic potential and the mechanism of action of a newly designed TCM formula, JinHuangJieDu (JHJD), which is composed of Lonicerae Japonicae Flos , Scutellariae Radix, Pogostemonis Herba , Angelicae Dahuricae Radix , Taraxaci Herba , and Menthae Haplocalycis Herba . Methods The inhibitory effects of JHJD on SARS-CoV-2 spike RBD binding to ACE2 and pseudovirus infection were assessed in vitro . Network pharmacology analyses predicted JHJD’s active ingredients, target genes, and enriched pathways. Key targets were validated in vitro for their role in mitigating virus-induced inflammation. Results JHJD effectively blocked wildtype and Omicron RBD binding to ACE2 and inhibited pseudovirus infection. Network analyses identified 375 ingredients, 338 targets (primarily in lung, stomach and spleen), and 63 significant pathways related to viral infection/inflammation. Intersectional analysis suggested CCND1 and HIF1A as pivotal genes and in vitr o validation confirmed that JHJD targets HIF-1α to attenuate inflammation. Conclusion JHJD, a de novo designed TCM formula, effectively inhibits SARS-CoV-2 entry with superior efficacy against Omicron sub-variants. The formulation has a dual mode of action including disruption of the RBD–ACE2 interaction and alleviation of virus-induced inflammation by modulating HIF-1α. These dual mechanisms establish JHJD as a promising therapeutic candidate for COVID-19.