Systematic Identification and Recognition Analysis of Beta1-Adrenergic Receptor Targeting Material Basis in Tibetan...

Bawei Chenxiang Powder (BWCP) is a Tibetan medicine formulation used to treat cardiovascular diseases, has demonstrated clinical efficacy though its material basis remains unclear. In clinical trial, β₁-adrenergic receptor (β₁-AR) has been recognized as a key therapeutic target in cardiovascular disease treatment. So, in this work, we engineered a fusion protein by linking the kinase domain of epidermal growth factor receptor (EGFR) to the C-terminus of β₁-AR, which was subsequently immobilized onto the ibrutinib-modified silica gels. Following column preparation by packing the β₁-AR immobilized silica into a stainless-steel column, the specificity and stability of β₁-AR column were evaluated through the retention behavior analysis of atenolol, esmolol, and metoprolol for characterization. Then, using the β₁-AR column combined with MS/MS, eight BWCP constituents targeting β₁-AR were screened and identified: 6-hydroxy-2-(2-phenylethyl) chromone, taxifolin, myristicin, costunolide, dehydrocostus lactone, gallic acid, 3,4-dihydroxybenzoic acid (DHBA), and catechin. The association constants of above screened compounds with immobilized β₁-AR were (6.75 ± 0.05) × 10⁴, (13.2 ± 0.60) × 10⁴, (3.82 ± 0.18) × 10⁴, (145.13 ± 1.50) × 10⁴, (199.77 ± 0.30) × 10⁴, (13.8 ± 0.04) × 10⁴, (9.30 ± 0.14) × 10⁴, (31.4 ± 0.76) × 10⁴ M^-1, respectively. Molecular docking results indicated that the compound-β₁-AR complexes stabilization primarily involves hydrogen bonding, van der Waals interactions, Pi-alkyl, and Pi-Pi stacking. A survey of DrugBank showed that catechin have well defined effects against cardiovascular diseases. DHBA and taxifolin are potential drug leads under experimental. These confirmed the reliability of the screened compounds by the immobilized β₁-AR. To our knowledge, this work establishes the first systematic identification of BWCP's bioactive components targeting β₁-AR, providing a pharmacological foundation for developing novel cardiovascular therapeutics derived from this traditional formulation.