title: Discovery of cytotoxic and pro-apoptotic compounds against leukemia cells: Tert-butyl-4-[(3-nitrophenoxy) methyl]-2,2-dimethyloxazolidine-3-carboxylate.
abstract: OBJECTIVE
  We evaluated biological activity in leukemia cells lines of R and S enantiomers of tert-butyl 4-[(3-nitrophenoxy)-methyl]-2,2-dimethyloxazolidine-3-carboxylate (BNDC).
  
  
  METHODS
  Cytotoxic activity was assessed by MTT assay. Flow cytometry assays were used to determined DNA fragmentation (Propidium Iodide-PI staining) and phosphatidylserine exposure (Annexin-V and PI staining). DNA condensation was evaluated by fluorescence microscopy using double-staining in leukemia cells (Hoechst and PI). Caspase activities were measured using Z-VAD-FMK, a non-selective caspase inhibitor, by flow cytometry and Z-DEVD-AMC, a selective caspase-3 substrate, by fluorescence spectrometry.
  
  
  RESULTS
  Both enantiomers displayed cytotoxic activity against leukemia cell lines (HL60, HL60.Bcl-2, HL60.Bcl-XL and Jurkat) with low toxicity against human peripheral blood mononuclear cell--PBMC based on IC50 values. In HL60 cell lines, compounds induce exposure of phosphatidylserine and DNA fragmentation, which could be blocked by pretreatment of cells with Z-VAD-FMK. Confirming this observation, both enantiomers induced caspase-3 activation. Additional analysis revealed an increased percentage of apoptotic cells (defined as those with fragmented nuclei and condensed chromatin) after treatment with compounds.
  
  
  CONCLUSIONS
  Taken together, the results indicate that BNDC compounds exhibited cytotoxic and pro-apoptotic activities and have a potential for developing a new class of anticancer drugs.
authors: Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, 31270-901, Belo Horizonte-MG, Brasil.
authors: Mauro C X
authors: MC
authors: Pinto
authors: Danielle F
authors: DF
authors: Dias
authors: Helen L
authors: HL
authors: Del Puerto
authors: Almir S
authors: AS
authors: Martins
authors: Andréa
authors: A
authors: Teixeira-Carvalho
authors: Olindo A
authors: OA
authors: Martins-Filho
authors: Bernard
authors: B
authors: Badet
authors: Philippe
authors: P
authors: Durand
authors: Ricardo J
authors: RJ
authors: Alves
authors: Elaine M
authors: EM
authors: Souza-Fagundes
mesh_terms: D000970:Antineoplastic Agents / Q000138:chemical synthesis* / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001562:Benzimidazoles; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation; D055808:Drug Discovery; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D018922:HL-60 Cells; D006801:Humans; D007202:Indicators and Reagents; D019169:Jurkat Cells; D007938:Leukemia / Q000188:drug therapy* / Q000473:pathology; D008856:Microscopy, Fluorescence; D010080:Oxazoles / Q000138:chemical synthesis* / Q000494:pharmacology*; D010718:Phosphatidylserines / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D013237:Stereoisomerism
chemical_list: D000970:Antineoplastic Agents; D001562:Benzimidazoles; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D010080:Oxazoles; D010718:Phosphatidylserines; D012333:RNA, Messenger; C568558:tert-butyl-4-((3-nitrophenoxy)methyl)-2,2-dimethyloxazolidine-3-carboxylate; D053148:Caspase 3; C017807:bisbenzimide ethoxide trihydrochloride