"Drug Discovery: Exploring the Anticancer Potential of new series of tricycle molecules, design, synthesis, and insights on their meccanism of action"

DRUG DISCOVERY: EXPLORING THE ANTICANCER POTENTIAL OF NEW SERIES OF TRICYCLES MOLECULES, DESIGN, SYNTHESIS, and INSIGHTS ON THEIR MECCANISM OF ACTION From chemical organic synthesis expertises ...to potential anticancer candidates Molecular Scaffolds under study to ISMN-Pa The pyrazolo[1,2-a]benzo[1,2,3,4]tetrazin-3-one derivatives PBTs have been previously designed as new alkylating agent because of their peculiar chemical behavior in soft acid environment towards charged or neutral nucleophiles. The tricycle is related to azolo-fused tetrazines such as Temozolomide, currently employed in cancer diseases. A preliminary NCI (USA) screening performed against more than 50 types of human tumor cell lines showed antiproliferative activity reaching in some cases sub-micromolar values [1] DESIGN: Recently, through the in silico VLAK protocol [2], a second series of new PBTS derivatives have been designed. Four derivatives have been selected, synthesized and evaluated on peculiar cellular events including proliferation, apoptosis induction and cell cycle perturbation [3]. Several of the new synthesized derivatives screened against apoptosis induction (HeLa) showed promising anticancer potential reaching in some cases activity in the micromolar range and arresting the cell cycle in the S phase. Useful candidates to develop as potential anticancer agents ! They act simultaneously on multiple biological pathways Further investigations on DNA /topoisomerase complex approached by Flow Linear Dicroism techniques Additional dichroic signals appear at higher wavelengths (300-360 nm and 400-520 nm). The occurrence of a dichroic signal at these wavelengths, where no contribution from DNA base pairs can be detected, indicates that this compound becomes oriented in the flow field, that is the formation of a molecular complex with the macromolecule Experimental validation on in vitro catalytic cycle of DNA/topoisomerase [4] Only M382 , M347 Derivatives inhibited the catalytic cycle of topoisomerase II, according to LD outcomes. These data suggest that the intercalation process could be the reason for the cell cycle phase arrest on S phase and inhibiting topoisomerase II catalytic cycle In silico investigations on DNA interaction REMARKS & PERSPECTIVES Taking into account all the chemical diversity points in the scaffold, along with the commercial availability of the reactants, the extension of the library of new compounds coupled with further in silico optimization studies could bring to improved results, so as to consider these compounds for preclinical studies [5] References [1] A.M. Almerico, F. Mingoia, P. Diana, P. Barraia, A. Lauria, A. Montalbano, G. Cirrincione, G. Dattolo, J. Med. Chem., 2005, 48, 2859-2866. [2] A. Lauria, M. Tutone, A.M. Almerico, Eur. J. Med. Chem., 2011, 46, 4274-4280. [3] F. Mingoia, C. Di Sano, F. Di Blasi, M. Fazzari, A. Martorana, A.M. Almerico, A. Lauria, Eur. J. Med Chem., 2013, 64, 345-356. [4] F. Mingoia et al 2016, manuscript in preparation. [5] This work was supported by RSTL. 654 2008 Francesco Mingoia.

From chemical organic synthesis expertises ...to potential anticancer candidates: Molecular Scaffolds currently under study to ISMN-Pa The pyrazolo[1,2-a]benzo[1,2,3,4]tetrazin-3-one derivatives PBTs have been previously  designed as new alkylating agent because of their peculiar chemical behavior in soft acid environment towards charged or neutral nucleophiles. The tricycle is related to azolo-fused tetrazines such as Temozolomide, currently employed in cancer diseases. A preliminary NCI (USA) screening performed against more than 50 types of human tumor cell lines showed antiproliferative activity reaching in some cases sub-micromolar values [1]. DESIGN: Recently, through the in silico VLAK protocol [2], a second series of new PBTS derivatives have been designed. Four derivatives have been selected, synthesized and evaluated on peculiar cellular events including proliferation, apoptosis induction and cell cycle perturbation [3]. Several of the new synthesized derivatives screened against apoptosis induction (HeLa) showed promising anticancer potential reaching in some cases activity in the micromolar range and arresting the cell cycle in the S phase. Useful candidates to develop as potential anticancer agents ! They act simultaneously on multiple biological pathways. Further investigations on DNA /topoisomerase complex approached by Flow Linear Dicroism techniques. Additional dichroic signals appear at higher wavelengths (300-360 nm and 400-520 nm). The occurrence of a dichroic signal at these wavelengths, where no contribution from DNA base pairs can be detected, indicates that this compound becomes oriented in the flow field, that is the formation of a molecular complex with the macromolecule. Experimental validation on in vitro catalytic cycle of DNA/topoisomerase [4] . Only M382 , M347 Derivatives inhibited the catalytic cycle of topoisomerase II, according to LD outcomes. These data suggest that the intercalation process could be the reason for the cell cycle phase arrest on S phase and inhibiting topoisomerase II catalytic cycle. In silico investigations on DNA interactions. REMARKS & PERSPECTIVES Taking into account all the chemical diversity points in the scaffold, along with the commercial availability of the reactants, the extension of the library of new compounds coupled with further in silico optimization studies could bring to improved results, so as to consider these compounds for preclinical studies [5]. References [1] A.M. Almerico, F. Mingoia, P. Diana, P. Barraia, A. Lauria, A. Montalbano, G. Cirrincione, G. Dattolo, J. Med. Chem., 2005, 48, 2859-2866. [2] A. Lauria, M. Tutone, A.M. Almerico, Eur. J. Med. Chem., 2011, 46, 4274-4280. [3] F. Mingoia, C. Di Sano, F. Di Blasi, M. Fazzari, A. Martorana, A.M. Almerico, A. Lauria, Eur. J. Med Chem., 2013, 64, 345-356. [4] F. Mingoia et al 2016, manuscript in preparation. [5] This work was supported by RSTL. 654 2008 Francesco Mingoia.