BIBR 1532

331, 36

0 to 80 uM

BIBR 1532 exhibits an non-competitive inhibitory effect on telomerase activity. [1] In JVM13 leukemia cell line, BIBR 1532 shows an antiproliferative effect in a dose-dependent range with IC50 of 52 uM, and similar results are also observed in other leukemia cell lines including Nalm-1, HL-60, and Jurkat. In addition, BIBR 1532 results in a direct, antiproliferative effect on acute myeloid leukemia (AML) with IC50 of 56 uM without affecting the proliferative capacity of normal hematopoietic progenitor cells. [2], BIBR 1532 (2.5 uM) reduces colony-forming ability, and induces telomere length shortening as well as chemotherapeutic sensitization by inhibiting telomerase activity in MCF-7/WT and melphalan-resistant MCF-7/MlnR cell lines. [3] In T-cell prolymphocytic leukemia (T-PLL), BIBR 1532 shows selective cytotoxic effects in a dose-dependent manner and BIBR 1532-treated cells also demonstrates nuclear condensation and formation of apoptotic bodies morphologically compatible with apoptosis. [4] A recent study shows that combination treatment of BIBR 1532 and chemotherapeutic agents carboplatin results in a potential synergy for eliminateing ovarian cancer spheroid-forming cells in ES2, SKOV3, and TOV112D cell lines. [5]

Conventional Telomerase Assay, For the direct telomerase assay with the endogenous telomerase, 10 uL of telomerase-enriched extract is mixed with different concentrations of BIBR1532 in a final volume of 20 uL. After 15-minute preincubation on ice, 20 uL of the reaction mixture is added, and the reaction is initiated by transferring the tubes to 37 C. The final concentrations in the reaction mixture are 25 mM Tris-Cl (pH 8.3), 1 mM MgCl2, 1 mM EGTA, 1 mM dATP, 1 mM dTTP, 6.3 uM cold dGTP, 15 uCi [alpha-32P]dGTP (3000 Ci/mmol, NEN), 1.25 mM spermidine, 10 units of RNasin, 5 mM 2-mercaptoethanol, and 2.5 uM TS-primer (5'-AATCCGTCGAGCAGAGTT). For the recombinant enzyme, 1–7 uL of affinity-purified telomerase (containing less than 0.025 um hTERT) are assayed in a final volume of 40 uL containing 50 mM Tris acetate (pH 8.5), 50 mM KCl, 1 mM MgCl2, 1 mM spermidine, 5 mM 2-mercaptoethanol, 1 mM dATP, 1 mM dTTP, 2.5 uM dGTP, 15 uCi of [alpha-32P]dGTP (3000 Ci/mmol) and 2.5 um (TTAGGG)3. The reaction is initiated incubation at 37 C for 2 hours and stopped by addition of 50 uL of RNase mix (0.1 mg/mL RNaseA-100 u/mL RNaseT1 in 10 mM Tris-Cl (pH 8.3) and 20 mm EDTA) and incubation for 20 min at 37 C. Samples are deproteinated by adding 50 uL of 0.3 mg/m proteinase K in 10 mM Tris-Cl (pH 8.3) and 0.5% w/v SDS, for a 30-minute incubation at 37 C. DNA is recovered by phenol extraction and ethanol precipitation, and the extension products are analyzed on an 8% (endogenous telomerase) or 12% (recombinant telomerase) polyacrylamide-urea gel. Dried gels are exposed to a Kodak phosphorimager screen, and the results are analyzed.

DMSO 66 mg/mL, Water <1 mg/mL, Ethanol 3 mg/mL

(E)-2-(3-(naphthalen-2-yl)but-2-enamido)benzoic acid