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Background: Adenosine is a metabolite that suppresses antitumor immune response of T and NK cells via extracellular binding to the two subtypes of adenosine-2 receptors, A 2 ARs. While blockade of the A 2A ARs subtype effectively rescues lymphocyte activity, with four A 2A AR antagonists currently in anticancer clinical trials, less is known for the therapeutic potential of the other A 2B AR blockade within cancer immunotherapy. Recent studies suggest the formation of A 2A AR/A 2B AR dimers in tissues that coexpress the two receptor subtypes, where the A 2B AR plays a dominant role, suggesting it as a promising target for cancer immunotherapy., Methods: We report the synthesis and functional evaluation of five potent A 2B AR antagonists and a dual A 2A AR/A 2B AR antagonist. The compounds were designed using previous pharmacological data assisted by modeling studies. Synthesis was developed using multicomponent approaches. Flow cytometry was used to evaluate the phenotype of T and NK cells on A 2B AR antagonist treatment. Functional activity of T and NK cells was tested in patient-derived tumor spheroid models., Results: We provide data for six novel small molecules: five A 2B AR selective antagonists and a dual A 2A AR/A 2B AR antagonist. The growth of patient-derived breast cancer spheroids is prevented when treated with A 2B AR antagonists. To elucidate if this depends on increased lymphocyte activity, immune cells proliferation, and cytokine production, lymphocyte infiltration was evaluated and compared with the potent A 2A AR antagonist AZD-4635. We find that A 2B AR antagonists rescue T and NK cell proliferation, IFNγ and perforin production, and increase tumor infiltrating lymphocytes infiltration into tumor spheroids without altering the expression of adhesion molecules., Conclusions: Our results demonstrate that A 2B AR is a promising target in immunotherapy, identifying ISAM-R56A as the most potent candidate for A 2B AR blockade. Inhibition of A 2B AR signaling restores T cell function and proliferation. Furthermore, A 2B AR and dual A 2A AR/A 2B AR antagonists showed similar or better results than A 2A AR antagonist AZD-4635 reinforcing the idea of dominant role of the A 2B AR in the regulation of the immune system., Competing Interests: Competing interests: None declared, (© Author(s) (or their employer(s)) 2022. Re-use permitted under CC BY. Published by BMJ.)

We here report on our continued studies of ligands binding to the promising drug target angiotensin II type 2 receptor (AT 2 R). Two series of compounds were synthesized and investigated. The first series explored the effects of adding small substituents to the phenyl ring of the known selective nonpeptide AT 2 R antagonist C38 , generating small but significant shifts in AT 2 R affinity. One compound in the first series was equipotent to C38 and showed similar kinetic solubility, and stability in both human and mouse liver microsomes. The second series was comprised of new bicyclic derivatives, amongst which one ligand exhibited a five-fold improved affinity to AT 2 R as compared to C38 . The majority of the compounds in the second series, including the most potent ligand, were inferior to C38 with regard to stability in both human and mouse microsomes. In contrast to our previously reported findings, ligands with shorter carbamate alkyl chains only demonstrated slightly improved stability in microsomes. Based on data presented herein, a more adequate, tentative model of the binding modes of ligand analogues to the prototype AT 2 R antagonist C38 is proposed, as deduced from docking redefined by molecular dynamic simulations.

The present study introduces a new strategy of selection of a maximum diversity sample of n compounds from N available in a molecular database. This strategy can be useful in pharmacological screening, combinatorial chemistry or parallel synthesis planning. It consists of first describing the compounds by means of parameters derived from quantum mechanical computations (water solvation deltaG, benzene solvation deltaG, octanol solvation deltaG, dipolar moment), as well as standard molecular parameters such as solvent-accessible surface area and molecular weight. Solvation parameters are used because of the importance of this phenomenon in the pharmacological behaviour. Redundant information in the description of the compounds is eliminated by using principal components (PC) instead of the original descriptors. Based on the similarity between the N compounds in the PC space, they are classified into n groups by k-means cluster analysis. The compounds that are nearest to the centroid of each cluster constituted the maximum diversity sample. When practical difficulties exist for the use of one of the proposed compounds, another also close to the cluster centroid can substitute for it. This strategy has been tested in the selection of a sample of 50 amines from the 923 available in the Aldrich catalogue. The results have been contrasted with those obtained from an optimal, distance-based experimental design, resulting in an 86% of agreement between both approaches. An R(2)-like diversity coefficient has been used to assess the quality of the proposed solutions.