{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:30:13Z","timestamp":1785407413290,"version":"3.56.0"},"reference-count":52,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T00:00:00Z","timestamp":1776124800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100012166","name":"National Key R&amp;D Program of China","doi-asserted-by":"publisher","award":["2025YFC3508601"],"award-info":[{"award-number":["2025YFC3508601"]}],"id":[{"id":"10.13039\/501100012166","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100016350","name":"Sichuan Provincial Administration of Traditional Chinese Medicine","doi-asserted-by":"publisher","award":["\/"],"award-info":[{"award-number":["\/"]}],"id":[{"id":"10.13039\/501100016350","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100019069","name":"Chinese Academy of Engineering","doi-asserted-by":"publisher","award":["2025-XZ-116"],"award-info":[{"award-number":["2025-XZ-116"]}],"id":[{"id":"10.13039\/501100019069","id-type":"DOI","asserted-by":"publisher"}]},{"award":["2025-XZ-116"],"award-info":[{"award-number":["2025-XZ-116"]}],"id":[{"id":"https:\/\/ror.org\/00z3yke57","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>Natural products represent an important reservoir for GPCR ligand discovery. In this study, we established an integrated workflow combining virtual screening, biophysical validation, functional signaling assays, and transcriptomic profiling to identify reticuline, a dopamine-derived intermediate from the genus of Stephania, as a potential agonist of dopamine D5 receptor (D5R). Molecular docking revealed that most dopamine-derived compounds along the BIA synthetic pathway exhibit predicted binding affinities for the D5R that are lower than that of dopamine. As expected, the reticuline\u2013D5R complex has a favorable predicted binding affinity of \u22127.9 kcal\/mol. As for binding validation, direct interaction between reticuline and D5R was experimentally confirmed using cell membrane chromatography (CMC) and bio-layer interferometry (BLI), yielding a dissociation constant of 1.07 \u03bcM. cAMP assay demonstrated that reticuline activates D5R-mediated Gs-cAMP increasement in a concentration-responsive manner, which exhibits agonist-like activity with an EC50 value of 0.07 \u03bcM. The transcriptomic profiling further revealed that reticuline treatment induces transcriptional reprogramming in D5R-overexpressing cells, with enrichment of pathways related to ribosome biogenesis, mitochondrial oxidative phosphorylation, and neurodegenerative diseases. In summary, this study demonstrates that reticuline acts as a potential D5R agonist and highlights a systematic natural product-GPCR discovery strategy integrating computational prediction, experimental validation, and transcriptome-level mechanistic exploration.<\/jats:p>","DOI":"10.3390\/molecules31081285","type":"journal-article","created":{"date-parts":[[2026,4,14]],"date-time":"2026-04-14T15:47:02Z","timestamp":1776181622000},"page":"1285","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Virtual and CMC-Based Screening Identified Reticuline, an Intermediate of BIA Biosynthesis, as a Potential Agonist of D5R"],"prefix":"10.3390","volume":"31","author":[{"given":"Jing","family":"Mo","sequence":"first","affiliation":[{"name":"College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-3482-5946","authenticated-orcid":false,"given":"Zhihao","family":"Sun","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guoqing","family":"Xu","sequence":"additional","affiliation":[{"name":"Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guichun","family":"Zhang","sequence":"additional","affiliation":[{"name":"Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhuangyuan","family":"Xie","sequence":"additional","affiliation":[{"name":"Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1347-3056","authenticated-orcid":false,"given":"Jinghao","family":"Zhao","sequence":"additional","affiliation":[{"name":"Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Go Pei","family":"Heng","sequence":"additional","affiliation":[{"name":"M Kandiah Faculty of Medicine and Health Sciences, Universiti Tunku Abdul Rahman, Kuala Lumpur 50744, Malaysia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhaotong","family":"Cong","sequence":"additional","affiliation":[{"name":"Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9629-6808","authenticated-orcid":false,"given":"Liang","family":"Leng","sequence":"additional","affiliation":[{"name":"Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shilin","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China"},{"name":"Innovative Institute of Chinese Medicine and Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"4011","DOI":"10.1016\/j.apsb.2022.08.022","article-title":"New Opportunities and Challenges of Natural Products Research: When Target Identification Meets Single-Cell Multiomics","volume":"12","author":"Zhu","year":"2022","journal-title":"Acta Pharm. 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