Label-free cell phenotypic profiling of histamine H4R receptor and discovery of non-competitive H4R antagonist from natural products

Hongming Tang; Tao Hou; Han Zhou; Han Liao; Fangfang Xu; Xiaomin Xie; Wenjie Yuan; Zhixin Guo; Yanfang Liu; Jixia Wang; Weijia Zhou; Xinmiao Liang

doi:10.1016/j.bioorg.2024.107387

Label-free cell phenotypic profiling of histamine H4R receptor and discovery of non-competitive H4R antagonist from natural products

Bioorg Chem. 2024 Apr 18:147:107387. doi: 10.1016/j.bioorg.2024.107387. Online ahead of print.

Authors

Hongming Tang¹, Tao Hou², Han Zhou³, Han Liao⁴, Fangfang Xu⁵, Xiaomin Xie⁶, Wenjie Yuan⁷, Zhixin Guo⁸, Yanfang Liu⁹, Jixia Wang¹⁰, Weijia Zhou¹¹, Xinmiao Liang¹²

Affiliations

¹ College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China. Electronic address: 1810170010@qq.com.
² CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: houtao141x@dicp.ac.cn.
³ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: zhouhan418@dicp.ac.cn.
⁴ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China. Electronic address: liaohan@dicp.ac.cn.
⁵ Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: fangfangxu@dicp.ac.cn.
⁶ Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: xiexiaomin@icmsc.cn.
⁷ Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: yuanwenjie@jcmsc.cn.
⁸ College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China. Electronic address: guozhixin@dlou.edu.cn.
⁹ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: liuyanfang@dicp.ac.cn.
¹⁰ CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: jxwang@dicp.ac.cn.
¹¹ College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China. Electronic address: zhouweijia@dlou.edu.cn.
¹² CAS Key Laboratory of Separation Science for Analytical Chemistry, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian 116023, China; Ganjiang Chinese Medicine Innovation Center, Nanchang 330000, China. Electronic address: liangxm@dicp.ac.cn.

PMID: 38643561
DOI: 10.1016/j.bioorg.2024.107387

Abstract

Histamine 4 receptor (H4R), the most recently identified subtype of histamine receptor, primarily induces inflammatory reactions upon activation. Several H4R antagonists have been developed for the treatment of inflammatory bowel disease (IBD) and atopic dermatitis (AD), but their use has been limited by adverse side effects, such as a short half-life and toxicity. Natural products, as an important source of anti-inflammatory agents, offer minimal side effects and reduced toxicity. This work aimed to identify novel H4R antagonists from natural products. An H4R target-pathway model deconvoluted downstream G_i and MAPK signaling pathways was established utilizing cellular label-free integrative pharmacology (CLIP), on which 148 natural products were screened. Cryptotanshinone was identified as selective H4R antagonist, with an IC₅₀ value of 11.68 ± 1.30 μM, which was verified with Fluorescence Imaging Plate Reader (FLIPR) and Cellular Thermal Shift (CTS) assays. The kinetic binding profile revealed the noncompetitive antagonistic property of cryptotanshinone. Two allosteric binding sites of H4R were predicted using SiteMap, Fpocket and CavityPlus. Subsequent molecular docking and dynamics simulation indicated that cryptotanshinone interacts with H4R at a pocket formed by the outward interfaces between TM3/4/5, potentially representing a new allosteric binding site for H4R. Overall, this study introduced cryptotanshinone as a novel H4R antagonist, offering promise as a new hit for drug design of H4R antagonist. Additionally, this study provided a novel screening model for the discovery of H4R antagonists.

Keywords: Antagonist; Cellular label-free integrative pharmacology; Cryptotanshinone; Histamine 4 receptor; Natural products.