TY - JOUR
T1 - Cell-based and in-silico studies on the high intrinsic activity of two boron-containing salbutamol derivatives at the human β 2- adrenoceptor
AU - Soriano-Ursúa, Marvin A.
AU - McNaught-Flores, Daniel A.
AU - Nieto-Alamilla, Gustavo
AU - Segura-Cabrera, Aldo
AU - Correa-Basurto, José
AU - Arias-Montaño, José A.
AU - Trujillo-Ferrara, José G.
N1 - Funding Information:
Partially supported by Cinvestav, Conacyt (Consejo Nacional de Ciencia y Tecnología; Grants 49371M to J.A.A.M., 132353 to J.C.B. and J.T.F., and scholarships to S.U.M.A., M.F.D. and S.C.A.) and Secretaría de Investigación y Posgrado del Instituto Politécnico Nacional. Authors thank the Conacyt-INNOVAPyME Program Support (139391) provided to Instituto Politécnico Nacional and Laboratorio Médico Químico Biológico. Also, JCB received support from ICyTDF and Fundación Miguel Alemán. Finally, we thank the access to computational resources from Centro Nacional de Supercómputo for MD simulations in this study.
PY - 2012/1/15
Y1 - 2012/1/15
N2 - Salbutamol is a well-known β 2 adrenoceptor (β 2AR) partial agonist. We synthesized two boron-containing salbutamol derivatives (BCSDs) with greater potency and efficacy, compared to salbutamol, for inducing β 2AR-mediated smooth-muscle relaxation in guinea-pig tracheal rings. However, the mechanism involved in this pharmacological effect remains unclear. In order to gain insight, we carried out binding and functional assays for BCSDs in HEK-293T cells transfected with the human β 2AR (hβ 2AR). The transfected hβ 2AR showed similar affinity for BCSDs and salbutamol, but adenosine 3′,5′-cyclic phosphate (cAMP) accumulation induced by both BCSDs was similar to that elicited by isoproterenol and greater than that induced by salbutamol. The boron-containing precursors (boric and phenylboronic acids, 100 μM) had no significant effect on salbutamol binding or salbutamol-induced cAMP accumulation. These experimental results are in agreement with theoretical docking simulations on lipid bilayer membrane-embedded hβ 2AR structures. These receptors showed slightly higher affinity for BCSDs than for salbutamol. An essential change between putative active and inactive conformational states depended on the interaction of the tested ligands with the fifth, sixth and seventh transmembrane domains. Overall, these data suggest that BCSDs induce and stabilize conformational states of the hβ 2AR that are highly capable of stimulating cAMP production.
AB - Salbutamol is a well-known β 2 adrenoceptor (β 2AR) partial agonist. We synthesized two boron-containing salbutamol derivatives (BCSDs) with greater potency and efficacy, compared to salbutamol, for inducing β 2AR-mediated smooth-muscle relaxation in guinea-pig tracheal rings. However, the mechanism involved in this pharmacological effect remains unclear. In order to gain insight, we carried out binding and functional assays for BCSDs in HEK-293T cells transfected with the human β 2AR (hβ 2AR). The transfected hβ 2AR showed similar affinity for BCSDs and salbutamol, but adenosine 3′,5′-cyclic phosphate (cAMP) accumulation induced by both BCSDs was similar to that elicited by isoproterenol and greater than that induced by salbutamol. The boron-containing precursors (boric and phenylboronic acids, 100 μM) had no significant effect on salbutamol binding or salbutamol-induced cAMP accumulation. These experimental results are in agreement with theoretical docking simulations on lipid bilayer membrane-embedded hβ 2AR structures. These receptors showed slightly higher affinity for BCSDs than for salbutamol. An essential change between putative active and inactive conformational states depended on the interaction of the tested ligands with the fifth, sixth and seventh transmembrane domains. Overall, these data suggest that BCSDs induce and stabilize conformational states of the hβ 2AR that are highly capable of stimulating cAMP production.
KW - 7TM
KW - Allosteric modulation
KW - Beta adrenergic receptor agonists
KW - Boron
KW - Drug-receptor interaction
UR - http://www.scopus.com/inward/record.url?scp=84855798366&partnerID=8YFLogxK
U2 - 10.1016/j.bmc.2011.11.054
DO - 10.1016/j.bmc.2011.11.054
M3 - Artículo
C2 - 22182578
SN - 0968-0896
VL - 20
SP - 933
EP - 941
JO - Bioorganic and Medicinal Chemistry
JF - Bioorganic and Medicinal Chemistry
IS - 2
ER -