Abstract
The design of beta2 adrenoceptor (β2AR) agonists is attractive because of their wide-ranging applications in medicine, and the details of agonist interactions with β2AR are interesting because it is considered a prototype for G-protein coupled receptors. Preclinical studies for agonist development have involved biological assays with guinea pigs due to a similar physiology to humans. Boron-containing Albuterol derivatives (BCADs) designed as bronchodilators have improved potency and efficacy compared with their boron-free precursor on guinea pig β2ARs (gpβ2ARs), and two of the BCADs (BR-AEA and boronterol) conserve these features on cells expressing human β2ARs (hβ2ARs). The aim of this study was to test the BCAD Politerol on gpβ2ARs and hβ2ARs in vitro and in silico. Politerol displayed higher potency and efficacy on gpβ2AR than on hβ2AR in experimental assays, possible explanations are provided based on molecular modeling, and molecular dynamics simulations of about 0.25 µs were performed for the free and bound states adding up to 2 µs in total. There were slight differences, particularly in the role of the boron atom, in the interactions of Politerol with gpβ2ARs and hβ2ARs, affecting movements of transmembrane domains 5–7, known to be pivotal in receptor activation. These findings could be instrumental in the design of compounds selective for hβ2ARs.
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Acknowledgements
We thank Bruce Allan Larsen for reviewing the use of English in the manuscript. The authors are grateful for financial support and scholarships from Comisión de Operación y Fomento de Actividades Académicas, Secretaría de Investigación y Posgrado of the IPN (SIP-M1930), and Consejo Nacional de Ciencia y Tecnología (CONACyT CB235785). JCB thanks to CONACYT CB-254600, APN-782 and Proyecto INSIGNIA-IPN-2015. Also, we thank to Escuela Nacional de Ciencias Biológicas for sharing facilities for our projects with boron-containing compounds.
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MASU, MB and JGTF conceived of the presented idea. MASU, MB, CFHM, JCB developed the theory and performed the computations. MASU, MB, JGTF and JAAM verified the analytical methods. JAAM encouraged IST and RGR to investigate Cell-based assays and supervised the findings of this work. All authors discussed the results and contributed to the final manuscript.
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This information includes the structure of boron-containing Albuterol adducts tested in silico on guinea pig and human β2AR models (Suppl. Fig. 1), docking studies to validate the procedure (In Suppl. Fig. 2, results from radioligand binding assays on membranes from transfected CHO-K1 cells (Suppl. Fig. 3 and Suppl. Table 1); the estimated affinity values for tested ligands (Suppl. Fig. 4), the MD simulation box of the extracellular and transmembrane regions of hβ2AR and gpβ2AR systems (Suppl. Fig. 5) and detailed data from MD simulations (Suppl. Fig. 6, Suppl. Fig. 7, Suppl. Table 2-5). 1 (PDF 1008 kb)
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Soriano-Ursúa, M.A., Bello, M., Hernández-Martínez, C.F. et al. Cell-based assays and molecular dynamics analysis of a boron-containing agonist with different profiles of binding to human and guinea pig beta2 adrenoceptors. Eur Biophys J 48, 83–97 (2019). https://doi.org/10.1007/s00249-018-1336-9
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DOI: https://doi.org/10.1007/s00249-018-1336-9