Description
The knowledge of drug-receptor interactions is an important issue in the field of new drug design and development of novel therapies. The main aim of this project is to study the drug–receptor interactions using an analytical technique (affinity chromatography). The new chromatographic columns with β2 adrenergic receptor (β2-AR) and GPR55 immobilized onto a stationary phase will be obtained. The resulting columns will be used to estimate the affinities of tested compounds toward studied receptors (β2-AR and GPR55). This strategy for selective screening of the drugs is facile, cost-effective, time-saving, ease-of-operation and it is promising for extensive usage of study of the specific target biomolecules. The studied receptors are the members of G-protein coupled receptors (GPCRs), which are very important targets for pharmacological intervention. Thus, the obtained results will be applicable in many fields (medicine, pharmacy, chemistry, biomolecular modeling).
Summary of project results
The main aim of this project was to study the ligand–receptor interactions using a series of experimental and computational techniques. In particular, we used high-tech experimental methods (affinity chromatography, in vitro studies) and newly developed computational methodologies to study the interactions between drugs and two receptors: β2 adrenergic (β2-AR) and GPR55. β2-AR is significant for the asthma treatment whereas the role of GPR55 has not yet been fully recognized. We prepared several selective cellular membrane affinity chromatography columns (CMAC columns) immobilized with β2-AR and GPR55. The columns were produced in cooperation with the National Institute of Health (NIH, USA). The resulting columns were used to estimate the affinities of tested compounds toward studied receptors (β2-AR and GPR55). This strategy for selective screening of the potential drugs is facile, cost-effective, time-saving, ease-of-operation and promising for extensive usage of study of the specific target biomolecules (e.g. receptors). The project’s output has a character of both contribution to the knowledge about GPCRs structure and function as well as development of applicable technologies. In particular, we developed protocols capable to predict the ligand-protein binding affinities dependent on the ligand stereochemistry: (i) the chromatographic separation of beta-mimetic enantiomers by using CMAC columns immobilized with β2-AR and GPR55 receptors and (ii) the computational, stereoconfiguration-independent potential (SIP) method. The other results concern: (i) in vitro estimation of the affinities of fenoterol derivatives toward GPR55, (ii) investigation of the mechanism of interactions between functionally different ligands and β2-AR and GPR55 and (iv) construction of the pharmacophor for inhibition of GPR55 and design a series of the new antagonists of GPR55. Additional result was identifying the novel and potent antagonist of GPR55 - (R,R)-metoxynaphtylfenoterol. Moreover, some currently available beta-blockers were found to inhibit GPR55-mediated activation of pro-survival ERK-signaling. The studies were extended by additional in vitro studies that revealed that (R,R)-metoxynaphtylfenoterol: (i) inhibits of breast cancer cell growth; (ii) inhibits migration of melanoma cell lines in culture and reduces melanoma cell proliferation.
Summary of bilateral results