BXI-72

Cas No.:	23491-45-4
Chemical Name:	Hoechst 33258 trihydrochloride
Synonyms:	Hoechst 33258;Bisbenzimide;p-(5-(5-(4-Methyl-1-piperazinyl)-1H-2-benzimidazolyl)-1H-2-benzimidazolyl)phenol trihydrochloride;2'-(4-Hydroxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bibenzimidazole Trihydrochloride Hydrate[for Biochemical Research];BISBENZIMIDE H 33258;Bisbenzimide H33258 Trihydrochloride;p-[5-(4-methyl-1-piperazinyl)(2,5'-bi-1H-benzimidazol)-2'-yl]phenol trihydrochloride;BBIH;Bisbenzimide H 33258 Hydrate;Butyltin trichloride;Hoechst Stain;Pibenzimol HCl;BXI-72;HOE 33258;NSC334072;33258 Hoechst;Hoechst Dye 33258;Bisbenzimidazole trihydrochloride;Hoechst 33258 Trihydrochloride;MHT095273M;Hoechst 33258 (trihydrochloride);H 33258;Phenol, 4-[5-(4-methyl-1-piperazinyl)[2,5'-bi-1H-benzimidazol]-2'-yl]-, trihydrochloride;Phenol, 4-(5-(4-methyl-1-piperazinyl)(2,5'-bi-1H-benzimidazol)-2'-yl)-, trihydrochloride;Bisbenzimide H 33258 Fluorochrome trihydrochloride;Bisbenzimide H 3325;bisBenzimide H 33258 trihydrochloride;H 33258 trihydrochloride;Hoechst 33258 trihydrochloride
SMILES:	Cl[H].Cl[H].Cl[H].O([H])C1C([H])=C([H])C(=C([H])C=1[H])C1=NC2C([H])=C([H])C(=C([H])C=2N1[H])C1=NC2C([H])=C([H])C(=C([H])C=2N1[H])N1C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])C1([H])[H]
Formula:	C25H27Cl3N6O
M.Wt:	533.8805
Purity:	>98%
Sotrage:	2 years -20°C Powder, 2 weeks 4°C in DMSO, 6 months -80°C in DMSO

Description:	BXI-72(Hoechst 33258) is a fluorescent dye that emits blue fluorescence when bound to dsDNA.
Target:	IC50: 51.31±4.56 μM (HeLa cell), 32.43±3.27 μM (HL60 cell), 15.42 ± 2.16 μM (U937 cell)[1]
In Vitro:	Hoechst 33258, a fluorescent compound with a head-to-tail bis-benzimidazole structure, is initially found to be cytotoxic against L1210 murine leukemia. Hoechst 33258 is evaluated for their cytotoxicity against human tumor cell lines, which are cervix carcinoma cell line (HeLa), Human promyelocytic leukemia cell (HL60) and U937 cell Line. The IC50 determined in the case of HeLa, HL60 and U937 is 51.31±4.56, 32.43±3.27 and 15.42±2.16 μM for Hoechst 33258, respectively[1]. The cytotoxic property of Hoechst 33258 is investigated on a panel of seven tumour cell lines of different histological origin and Madine-Darby canine kidney (MDCK) normal cells. All cell lines, except MCF-7, exposed to Hoechst 33258 exhibit GI50 from 84×10-6 to 191.5×10-6 mol/dm3. Under the same experimental conditions, Hoechst 33258, used as a binder reference compound, stops the cell cycle in S phase and G0/G1[2].
Cell Assay:	Hoechst 33258 is prepared as stock solutions in highly pure water. Working solutions in a concentration range of 10-3-10-6 mol/dm3 are prepared prior to testing. Cytotoxic effects of Hoechst 33258 on tested cell lines are determined by the MTT assay. Cells are seeded in 96 micro well flat bottom plates at a concentration of 2×104 cells/mL and left overnight in the CO2 incubator allowing them to attach to the plate surface. Growing medium is replaced with compound supplemented or control medium and incubated for 72 h. Fresh medium with 5 mg/mL of MTT is added onto cells and incubated for 4 h at 37°C. Upon media removal, water insoluble MTT-formazan crystals formed inside the living cells are dissolved in DMSO and the absorbance at 570 nm proportional to the number of living cells is measured on an Elisa Microplate Reader. All experiments are performed at least three times in triplicates.The GI50 value, defined as the compound concentration (μM) leading to cellular growth inhibition by 50%, is calculated and used as a parameter to compare cytotoxicity among the compounds[2].
References:	[1]. Wang XJ, et al. Newly synthesized bis-benzimidazole derivatives exerting anti-tumor activity through inductionof apoptosis and autophagy. Bioorg Med Chem Lett. 2012 Oct 1;22(19):6297-300. [2]. Stolić I, et al. Synthesis, DNA/RNA affinity and antitumour activity of new aromatic diamidines linked by 3,4-ethylenedioxythiophene. Eur J Med Chem. 2011 Feb;46(2):743-55.