Interação de drogas de recreação com ctDNA simulando condições fisiológicas

Abstract

There are recreational drugs that do not have published preclinical safety data regarding their toxicity. This may be related to both the difficulty in obtaining sufficient quantities of drug samples and the difficulty in obtaining a judicial release to carry out these studies. One example is the derivatives of NBOMe, this name comes from the substituent "N-benzylmethoxy" (the methoxy being written "OMe"), which have a variety of structures, are synthetic hallucinogens, phenethylamine derivatives and widely used as LSD (Lysergsäurediethylamid - lysergic acid diethylamide) which is one of the most potent hallucinogenic substances known. 25H-NBOMe and 25H-NBOH are examples of these drugs, but their physiological and toxicological properties are not known. In the studies employing molecular fluorescence, it was observed that the analyzed compounds showed emission at 327 nm when excited. at 260 nm. Thus, due to the variation in the intrinsic fluorescence of each compound, It is possible to calculate the Stern-Volmer constant (KSV) and the binding constant (Kb). The constants were calculated in the range of 23 to 37ºC, presenting KSV in the order of 103 M-1 and Kb in the order of 104 M-1, which decreased as a function of the temperature increase. The preferred mechanism of quenching the fluorescence of the compounds by ctDNA was classified as static quenching, since the Kq> 2.0 × 1010 M-1 s-1. By means of competition studies with seven commercial probes (ethidium bromide, acridine orange, DAPI, orange thiazole, berberine, hoechst and methyl green) with established dates, it was possible to infer which these compounds interact with ctDNA via the major groove. The thermodynamic studies indicated that the interaction with ctDNA is spontaneous and that the main types of interactions that occur are hydrogen bonds and van der Waals force (H <0 and S <0). Finally, using 1H NMR studies the epitope of the molecules was established in the presence of ctDNA. In the case of 25H-NBOH, the region containing the phenol was the one that most suffered displacement in the presence of the macromolecule, which is an indication that this is the portion of the molecule that most interacts with the ctDNA. In addition, molecular docking indicated the regions of interaction and what are the main interactions that stabilize the complex and are consistent with the experimental studies.Thus, this study presents evidence of the interaction between recreational drugs such as 25H-NBOMe and 25H-NBOH with DNA, which may lead to possible genotoxic effects.