Cytotoxic Effects of Dimorfolido-N-Trichloroacetylphosphorylamide and Dimorfolido-N-Benzoylphosphorylamide in Combination with C60 Fullerene on Leukemic Cells and Docking Study of Their Interaction with DNA

Abstract

Dimorfolido-N-trichloroacetylphosphorylamide (HL1) and dimorfolido-N-benzoylphosphorylamide (HL2) as representatives of carbacylamidophosphates were synthesized and identified by the methods of IR, 1H, and 31P NMR spectroscopy. In vitro HL1 and HL2 at 1 mM concentration caused cell specific and time-dependent decrease of leukemic cell viability. Compounds caused the similar gradual decrease of Jurkat cells viability at 72 h (by 35%). HL1 had earlier and more profound toxic effect as compared to HL2 regardless on leukemic cell line. Viability of Molt-16 and CCRF-CEM cells under the action of HL1 was decreased at 24 h (by 32 and 45%, respectively) with no substantial further reducing up to 72 h. Toxic effect of HL2 was detected only at 72 h of incubation of Jurkat and Molt-16 cells (cell viability was decreased by 40 and 45%, respectively).It was shown that C60 fullerene enhanced the toxic effect of HL2 on leukemic cells. Viability of Jurkat and CCRF-CEM cells at combined action of C60 fullerene and HL2 was decreased at 72 h (by 20 and 24%, respectively) in comparison with the effect of HL2 taken separately.In silico study showed that HL1 and HL2 can interact with DNA and form complexes with DNA both separately and in combination with C60 fullerene. More stable complexes are formed when DNA interacts with HL1 or C60 + HL2 structure. Strong stacking interactions can be formed between HL2 and C60 fullerene. Differences in the types of identified bonds and ways of binding can determine distinction in cytotoxic effects of studied compounds.

Water-Soluble Pristine C60 Fullerene Inhibits Liver Alterations Associated with Hepatocellular Carcinoma in Rat

“…However, if applied in doses less than 10–20 µg/mL (depending on cell line) C 60 fullerene didn’t increase the number of apoptotic and pre-apoptotic cells [ 46 ], being, therefore, relatively low-toxic. Low cytotoxicity of C 60 fullerene against normal and neoplastic cells was also demonstrated in [ 13 , 47 ]. Thus, C 60 fullerene did not affect leukemic cell viability when applying at 3.6–144 µg/mL (CCRF-CEM, Jurkat, Molt-16 cells), IC 50 for HEK293 cells was equal to 383.4 µg/mL.…”

C60 Fullerene Effects on Diphenyl-N-(trichloroacetyl)-amidophosphate Interaction with DNA In Silico and Its Cytotoxic Activity Against Human Leukemic Cell Line In Vitro

“…We have used two versions of molecular modeling of HL, C 60 fullerene and DNA interaction, which were proposed in [ 16 ] and proved to be useful for interpretation of MD simulation results. We have used 1XRW PDB structure of DNA molecule in the version, when initially HL and then C 60 molecule intercalate into DNA (HL+C 60 ) and 2MIW PDB structure of DNA molecule - in the version, when initially C 60 molecule and then HL intercalate into DNA(C 60 +HL).…”

“…5 ). Recently, we have shown that another CAPh representative dimorfolido-N-trichloracetylphosphorylamid caused 50% decrease of CCRF-CЕM cells viability at 72 h in concentration 1 mM [ 16 ]. The comparative analysis of these data demonstrates that introduction of phenoxy instead of morfolido groups into the structure of CAPh derivative allowed to decrease by two orders its effective toxic concentration against leukemic cells.…”

“…Previously, we have shown a significant toxic effect of dimorfolido-N-trichloroacetylphosphoramide against human leukemic cells of different origin, but its effective concentration was high and no enhancement of toxicity after combined action with C 60 fullerene was observed [ 16 ]. We assume that introduction of flexible of –P(O)(OC 6 H 5 ) groups instead of morfolido groups into CAPh derivative contributed to lowering of its toxic concentration against human leukemic cells ensuring its effective interaction with C 60 molecule and DNA.…”

“…Recently, we have confirmed that CAPh representative dimethyl-N-(benzoyl)-amidophosphate used in millimolar concentrations range decreased the viability of leukemic L1210 cells and that its toxic effect was facilitated by C 60 fullerene [ 15 ]. We have shown also that introduction of additional aromatic substituents and electronegative CCl 3 group into CAPh structure resulted in enhancement of its toxicity [ 16 ]. Thus, significant toxic effect of dimorfolido-N-trichloroacetylphosphoramide was shown against human leukemic cells of different origin, but its effective concentration was still high and no enhancement of toxicity after combined action with C 60 fullerene was observed.…”