Cell-based assays and molecular dynamics analysis of a boron-containing agonist with different profiles of binding to human and guinea pig beta2 adrenoceptors
- 40 Downloads
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.
KeywordsBoron GPCR Cell-based assays Adrenoceptor Comparative physiology Pharmacology
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.
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.
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interest.
- Bai Q, Shao Y, Pan D, Zhang Y, Liu H, Yao X (2014) Search for β2 adrenergic receptor ligands by virtual screening via grid computing and investigation of binding modes by docking and molecular dynamics simulations. PLoS One 9(9):e107837. https://doi.org/10.1371/journal.pone.0107837 CrossRefPubMedPubMedCentralGoogle Scholar
- DeLano WL (2002) The PyMOL molecular graphics system. DeLano Scientific, San Carlos, CA. http://www.pymol.org
- DeVree BT, Mahoney JP, Vélez-Ruiz GA, Rasmussen SG, Kuszak AJ, Edwald E, Fung JJ, Manglik A, Masureel M, Du Y, Matt RA, Pardon E, Steyaert J, Kobilka BK, Sunahara RK (2016) Allosteric coupling from G protein to the agonist-binding pocket in GPCRs. Nature 535(7610):182–186. https://doi.org/10.1038/nature18324 CrossRefPubMedPubMedCentralGoogle Scholar
- Dickson CJ, Hornak V, Velez-Vega C, McKay DJ, Reilly J, Sandham DA, Shaw D, Fairhurst RA, Charlton SJ, Sykes DA, Pearlstein RA, Duca JS (2016) Uncoupling the structure-activity relationships of β2Adrenergic receptor ligands from membrane binding. J Med Chem 59(12):5780–5789. https://doi.org/10.1021/acs.jmedchem.6b00358 CrossRefPubMedGoogle Scholar
- Frisch MJ, Trucks GW, Schlegel HB et al (2003) Gaussian03, Revision C.02. Gaussian Inc., Pittsburgh, PAGoogle Scholar
- Lamichhane R, Liu JJ, Pljevaljcic G, White KL, van der Schans E, Katritch V, Stevens RC, Wüthrich K, Millar DP (2015) Single-molecule view of basal activity and activation mechanisms of the G protein-coupled receptor β2AR. Proc Natl Acad Sci USA. 112(46):14254–14259. https://doi.org/10.1073/pnas.1519626112 CrossRefPubMedGoogle Scholar
- Nygaard R, Zou Y, Dror RO, Mildorf TJ, Arlow DH, Manglik A, Pan AC, Liu CW, Fung JJ, Bokoch MP, Thian FS, Kobilka TS, Shaw DE, Mueller L, Prosser RS, Kobilka BK (2013) The dynamic process of β(2) adrenergic receptor activation. Cell 152(3):532–542. https://doi.org/10.1016/j.cell.2013.01.008 CrossRefPubMedPubMedCentralGoogle Scholar
- Soriano-Ursúa MA, Valencia-Hernández I, Arellano-Mendoza MG, Correa-Basurto J, Trujillo-Ferrara JG (2009a) Synthesis, pharmacological and in silico evaluation of 1-(4-di-hydroxy-3,5-dioxa-4-borabicyclo[4.4.0]deca-7,9,11-trien-9-yl)-2-(tert-butylamino) ethanol, a compound designed to act as a beta2 adrenoceptor agonist. Eur J Med Chem 44:2840–2846. https://doi.org/10.1016/j.ejmech.2008.12.016 CrossRefPubMedGoogle Scholar
- Soriano-Ursúa MA, Trujillo-Ferrara J, Correa-Basurto J (2009b) Homology modeling and flex-ligand docking studies on the guinea pig β2 adrenoceptor: structural and experimental similarities/differences with the human β2. J Mol Model 15:1203–1211. https://doi.org/10.1007/s00894-009-0480-7 CrossRefPubMedGoogle Scholar
- Soriano-Ursúa MA, Correa-Basurto J, Valencia-Hernández I, Amezcua-Gutiérrez MA, Padilla-Martínez II, Trujillo-Ferrara JG (2010) Design, synthesis and in vitro evaluation of (R)-4-(2-(tert-butylamino)-1-hydroxyethyl)-2-(hydroxymethyl)phenyl hydrogen phenylboronate: a novel Albuterol derivative with high intrinsic efficacy on the β2 adrenoceptor. Bioorg Med Chem Lett 20:5623–5629. https://doi.org/10.1016/j.bmcl.2010.08.040 CrossRefPubMedGoogle Scholar
- Soriano-Ursúa MA, McNaught-Flores DA, Nieto-Alamilla G, Segura-Cabrera A, Correa-Basurto J, Arias-Montaño JA, Trujillo-Ferrara JG (2012) Cell-based and in silico studies on the high intrinsic activity of two boron-containing salbutamol derivatives at the human β2-adrenoceptor. Bioorg Med Chem 20(2):933–941. https://doi.org/10.1016/j.bmc.2011.11.054 CrossRefPubMedGoogle Scholar
- Soriano-Ursúa MA, Arias-Montaño JA, Correa-Basurto J, Hernández-Martínez CF, López-Cabrera Y, Castillo-Hernández MC, Padilla-Martínez II, Trujillo-Ferrara JG (2015b) Insights on the role of boron containing moieties in the design of new potent and efficient agonists targeting the β2 adrenoceptor. Bioorg Med Chem Lett 25(4):820–825. https://doi.org/10.1016/j.bmcl.2014.12.077 CrossRefPubMedGoogle Scholar