In Search of New Inhibitors of HIV-1 Replication: Synthesis and Study of 1-(2′-Deoxy-β-D-Ribofuranosyl)-1,2,4-Triazole-3-Carboxamide as a Selective Viral Mutagenic Agent

With the emergence of HIV strains resistant or cross-resistant to nearly all antiretroviral regimen, novel therapy approaches have to be considered. As a part of our current work on viral mutagenic compounds, we prepared 1-(2′ -deoxy-β-D-ribofuranosyl)-1,2,4-triazole-3-carboxamide (2′ -deoxy-ribavirin) and its 5′ -triphosphate derivative. The nucleoside mutagenic activity was evaluated on HIV-1 NL4-3 in CEMx174 cell culture. After 2.5 months, no reduction on HIV-1 viability was observed. On the other hand, in vitro experiments with purified HIV-1 RT demonstrated that the triphosphate analog can be incorporated opposite to several natural nucleosides.     

Elucidation of the Origin of NOEs And ROEs that Show the Hydration in the Minor and Major Grooves of DNA Duplex with ATTAAT Tract by a Combination of NOESY and ROESY Experiments

Abstract

A combination of NOESY and ROESY experiments (using ammonia as a catalyst across the pH range of 5 to 8.6) has given us a clear understanding regarding the origin of nOes that are attributed to the stereochemical location and the residence time of water in the major and the minor grooves of d⁵'(¹C²C³A⁴T⁵T⁶A⁷A⁸T⁹G¹⁰G)₂ ³' duplex Our conclusions are the following: (i) In the major groove, the presence of ammonia in the buffer does not influence on the process of exchange between bound and bulk water, (ii) It has been found that the observation of the bound water in the minor groove is a result of straight dipole-dipole effect at the physiological pH. (iii) The residence time of water near H2 of adenine (H2A) in the minor groove has been estimated to be in the range of 0.3–0.5ns, which is closer to the residence time of the bound water found on the surface of protein, (iv) The hydration pattern in the minor groove in the physiological pH, under our NMR measurement condition, is similar to the ones found in the X-ray structure, (v) It has been shown that at pH > 8.0 the nOe/rOe intensities of the water-H2A crosspeaks dramatically increase due to dipole-dipole and/or relayed magnetization transfer from H2A to water through ammonia catalyst.     

Investigation of the Synthesis of 2-Carbamoylmethyl-6-D-Ribofuranosylpyridine

Abstract

During the course of our study on the synthesis of new pyridine-C-nucleosides with potential anti-viral and/or anti-tumourai properties, the preparation of 2-carbamoylmethyl-6-β-D-ribofuranosylpyridine 1, by means of a lithiation reaction on 2-(2,3-0-isopropylidene-β D-nbofuranosyl)-6-methylpyridine 2, was investigated (Scheme I). The latter compound was prepared by treating 6-methyl- toluenesulphonic acid. 2-(& D-ribofuranosyl)pyridine 1 2 with 2.2-dimethoxypropane in the presence of p-     

Arenesulfonylethoxycarbonyl- a Set of Amino Protecting Groups for DNA and RNA Synthesis

Abstract

Phenyl-, 4-chlorophenyl- and 4-nitrophenylsufonylethoxycarbonyl groups have been reported for the first time as the exocyclic amino protecting groups in nucleoside chemistry. They are all stable under the standard conditions of manipulations in phosphotriester and phosphiteamidite chemistry, they are removable both under the alkaline hydrolytic conditions and also under the influence of non-nucleophilic tertiary bases. N³-Phenyl- and 4-toluenesulfonylethoxycarbonyl derivatives of uridine have been also prepared and characterized by ¹⁵N-NMR spectroscopy, their stabilities under different conditions have been tested.     

Achiral Nucleotide Analogs: 5,5-Disubstituted Pyrimidines Related to Barbituric Acid

Abstract

Achiral nucleotide analogs are of potential interest in origin of life studies. Four new 5,5-diphosphoethylpyrimidine analogs were prepared by condensation of urea or guanidine with a, a-di(2-[ethoxy-I-ethoxy]ethyl)malononitrile or a, a-di(2-[ethoxy-1-ethoxy]ethyl)ethyl cyanoacetate respectively, followed by removal of the protecting groups and bisphosphorylation. Oligomers of two potentially complementary analogs have been prepared. Interactions between these oligomers appear to be weak in aqueous solution.     

Solution NMR Conformational Analysis of the Potent Equilibrative Sensitive (ES) Nucleoside Transporter Inhibitor,S 6-(4-Nitrobenzyl)mercaptopurine Riboside (NBMPR)

Abstract

High resolution NMR analysis involving one-dimensional (1-D) ¹H and nuclear Overhauser (NOE) difference spectroscopy was applied to solutions of NBMPR in DMSO-d ₆. Coupling constants were obtained at different temperatures between 285 and 353 K, and used to analyze the rotamer preferences about the C-4′-C-5′ bond. The results revealed a rotamer distribution about the χ tortion angle that favors the high-anti range, a preponderance of the γ⁺ rotamer (at ～64 %) with respect to the γ torsion angle, and a higher population of the south (S) conformer, which was favored by as little as the 4 % to as much as 31 % over the north (N) conformer as calculated by the program PSEUROT 6.2. The high-anti glycosidic torsion orientation appears to be the major conformational difference between the solution structure of NBMPR determined in this study and the structure previously observed in the solid state.     

Multicoloured greenbeards,bacteriocin diversity and the rock‐paper‐scissors game

Greenbeard genes identify copies of themselves in other individuals and cause their bearer to behave nepotistically towards those individuals. Bacterial toxins (bacteriocins) exemplify the greenbeard effect because producer strains carry closely linked genes for immunity, such that toxicity is limited to nonproducer strains. Bacteriocin producers can be maintained in a dynamic polymorphism, known as rock‐paper‐scissors (RPS) dynamics, with immune and susceptible strains. However, it is unclear whether and how such dynamics will be maintained in the presence of multiple toxin types (multiple beard ‘colours’). Here, we analyse strain dynamics using models of recurrent patch colonization and population growth. We find that (i) polymorphism is promoted by a small number of founding lineages per patch, strong local resource competition and the occurrence of mutations; (ii) polymorphism can be static or dynamic, depending on the intensity of local interactions and the costs of toxins and immunity; (iii) the occurrence of multiple toxins can promote RPS dynamics; and (iv) strain diversity can be maintained even when toxins differ in toxicity or lineages can exhibit multitoxicity/multi‐immunity. Overall, the factors that maintain simple RPS dynamics can also promote the coexistence of multiple toxin types (multiple beard colours), thus helping to explain the remarkable levels of bacteriocin diversity in nature. More generally, we contrast these results with the maintenance of marker diversity in genetic kin recognition.     

Binding of thiazolidinediones to the endoplasmic reticulum protein nutrient-deprivation autophagy factor-1

Nutrient-deprivation autophagy factor-1 (NAF-1, miner1; gene cisd2) is part of the [2Fe-2S]-containing protein family which includes mitoNEET (gene cisd1) and MiNT (miner2; gene cisd3). These proteins are redox active and are thought to play an important role in cellular energy homeostasis with NAF-1 playing a critical role in calcium regulation and aging. To date, no studies have investigated potential ligand interaction with NAF-1. Here we show that the thiazolidinediones pioglitazone and rosiglitazone along with the mitoNEET ligand, NL-1, bind to NAF-1 with low micromolar affinities. Further, we show that overexpression of NAF-1 in hepatocellular carcinoma (HepG2) cells reduces inhibition of mitochondrial respiration by pioglitazone. Our findings support the need for further efforts of the rational design of selective NAF-1 ligands.