ABSTRACT
AIM: No antiviral medications are currently approved to treat enterovirus (EV)-associated disease or prevent EV infection. METHODS: In this study, a series of probenecid derivatives were designed via a rational strategy and synthesized to obtain more potent anti-EV agents. RESULTS: Compounds 8 and 24 exhibited the most potent activity against EV D68 and A71, with half maximal effective concentration (EC50) values of 2.49/2.09 and 2.59/2.41 µM, respectively, and revealed a broad inhibition spectrum toward other EV strains, with high selectivity indices. Additionally, compounds 8 and 24 showed good stability in rat serum, with half-lives of 48.39 and 60.26 min, respectively. CONCLUSION: Compounds 8 and 24 are the promising candidates for the development of new agents against EV D68 and A71 viruses.
Subject(s)
Antiviral Agents/chemical synthesis , Drug Design , Enterovirus/drug effects , Probenecid/chemical synthesis , Animals , Antiviral Agents/pharmacokinetics , Cell Line , Cell Survival/drug effects , Drug Evaluation, Preclinical , Drug Stability , Enterovirus Infections , Humans , Molecular Docking Simulation , Molecular Structure , Probenecid/analogs & derivatives , Probenecid/pharmacokinetics , Rats , Structure-Activity RelationshipABSTRACT
Novel Probenecid-based amide derivatives, incorporating different natural amino acids, were synthesized and assayed to test their effect on the human carbonic anhydrase (hCA, EC 4.2.1.1) transmembrane isoforms hCA IX and XII over the ubiquitous isoforms hCA I and II. Most of them presented a complete loss of hCA II inhibition (K(i)s > 10,000 nM) and strong inhibitory activity against hCA IX and XII in the nanomolar range with respect to the parent compound. A residual activity against hCA I was observed for some of them. These biological results have been explained by docking studies within the active sites of the four studied human carbonic anhydrases (with or without the zinc-bound water) and helped us to better comprehend the rationale behind the design of tertiary sulfonamide compounds as potent but atypical inhibitors of specific isoforms of human carbonic anhydrase.
Subject(s)
Carbonic Anhydrase Inhibitors/chemistry , Carbonic Anhydrase Inhibitors/pharmacology , Carbonic Anhydrases/metabolism , Probenecid/analogs & derivatives , Probenecid/pharmacology , Antigens, Neoplasm/chemistry , Antigens, Neoplasm/metabolism , Carbonic Anhydrase I/antagonists & inhibitors , Carbonic Anhydrase I/chemistry , Carbonic Anhydrase II/antagonists & inhibitors , Carbonic Anhydrase II/chemistry , Carbonic Anhydrase II/metabolism , Carbonic Anhydrase IX , Carbonic Anhydrase Inhibitors/chemical synthesis , Carbonic Anhydrases/chemistry , Catalytic Domain , Humans , Molecular Docking Simulation , Probenecid/chemical synthesis , Structure-Activity RelationshipABSTRACT
Novel amide derivatives of Probenecid were synthesized and discovered to act as potent and selective inhibitors of the human carbonic anhydrase (hCA, EC 4.2.1.1) transmembrane isoforms hCA IX and XII. The proposed chemical transformation of the carboxylic acid into an amide group led to a complete loss of hCA I and II inhibition (Kis >10,000nM) and enhanced the inhibitory activity against hCA IX and XII, with respect to the parent compound (incorporating a COOH function). These promising biological results have been corroborated by molecular modelling studies within the active sites of the four studied human carbonic anhydrases, which enabled us to rationalize both the isoform selectivity and high activity against the tumor-associated isoforms hCA IX/XII.
Subject(s)
Antigens, Neoplasm/chemistry , Carbonic Anhydrase II/chemistry , Carbonic Anhydrase I/chemistry , Carbonic Anhydrase Inhibitors/chemical synthesis , Carbonic Anhydrases/chemistry , Probenecid/chemical synthesis , Carbonic Anhydrase IX , Carbonic Anhydrase Inhibitors/chemistry , Catalytic Domain , Crystallography, X-Ray , Enzyme Assays , Humans , Molecular Docking Simulation , Probenecid/analogs & derivatives , Protein Binding , Sensitivity and Specificity , Structure-Activity RelationshipSubject(s)
Kidney Tubules/metabolism , Probenecid/analogs & derivatives , Probenecid/metabolism , Animals , Binding, Competitive , Female , In Vitro Techniques , Liposomes/metabolism , Male , Phenolsulfonphthalein/metabolism , Probenecid/chemical synthesis , Protein Binding , Rabbits , Serum Albumin/metabolism , p-Aminohippuric Acid/metabolismABSTRACT
The uptake of cyclic analogues of probenecid by kidney cortical slices has been studied in detail, in order to obtain more information on the secretory system for these compounds. Both p-piperidyl sulfamyl benzoic acid and p-benzyl sulfamyl benzoic acid were accumulated against concentration gradient, by renal tissue under aerobic as well as anerobic conditions. PAH, phenol red and probenecid competitively inhibited the active accumulation of these compounds by kidney tissue. Aerobic uptake of probenecid analogues was stimulated by succinate and octanoate at low medium concentrations while inhibition of renal accumulation of these compounds occurred at higher concentrations. Both p-piperidyl and p-benzyl sulfamyl benzoic acids like probenecid strongly interact with kidney cortex homogenates. Binding of these cyclic analogues to various cellular constituents of homogenate was efficiently inhibited by probenecid. The binding affinity of probenecid and analogues for kidney tissue, phospholipid vesicles (liposomes) and human serum albumin increased in the order : p-piperidyl sulfamyl benzoic acid less than p-benzyl sulfamyl benzoic acid less than di-n-propyl sulfamyl benzoic acid (probenecid). By contrast to the view put forward by Beyer (1950 & 1954), the results presented in this paper established that probenecid analogues are the true substrates of renal organic anion transport system.
