BAX AGONIST, COMPOSITIONS, AND METHODS RELATED THERETO

摘要

To compare sensitivities of the compounds, A549 human lung cancer cells were treated with increasing concentrations (0, 1, 5, 10, 25 μM) of 2-(2-Nitro-fluoren-9-ylidenemethyl)-phenol (CYD-1-87) and derivatives for 48 h. The surviving cell fraction was determined using the sulforhodamine B (SRB) assay as described in (Vichai & Kirtikara, Nat Protoc 1, 1112-1116, 2006 hereby incorporated by reference). The sulforhodamine B (SRB) assay is used for cell density determination, based on the measurement of cellular protein content. The method described here has been optimized for the toxicity screening of compounds to adherent cells in a 96-well format. After an incubation period, cell monolayers are fixed with 10% (wt/vol) trichloroacetic acid and stained for 30 min, after which the excess dye is removed by washing repeatedly with 1% (vol/vol) acetic acid. The protein-bound dye is dissolved in 10 mM Tris base solution for OD determination at 510 nm using a microplate reader. The results are typically linear over a 20-fold range of cell numbers. CYD-2-11 has an IC50 of 1.93 μM, Derivative CYD-2-17 has an IC50 of 5.08 μM and CYD-2-13 has an IC50 of 5.91 μM. Data additional obtained from this assay is provided in FIG. 5. SMBA1 has an IC50 of 7.35 and CYD-4-61, 2-((3-((2-nitro-fluoren-9-ylidene)methyl)pyridin-2-yl)oxy)ethanamine has an IC50 of 0.026.

主权项

1. A method of treating lung cancer comprising administering an effective amount of a compound of Formula IB, to a subject in need thereof, or salt thereof wherein, Z is O, S, CH2, or NH; W is hydroxy, amino, alkylamino, dialkylamino, aryl, or heterocyclyl wherein W is optionally substituted with one or more R11; R1, R3, R4, R5, R6, R7, and R8 are each individually and independently hydrogen, alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R1, R3, R4, R5, R6, R7, and R8 are optionally substituted with one or more, the same or different R10; R2 is nitro or amino wherein R2 is optionally substituted with one or more, the same or different R10; R10 is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R10 is optionally substituted with one or more, the same or different R11; R11 is alkyl, halogen, nitro, cyano, hydroxy, amino, mercapto, formyl, carboxy, carbamoyl, alkoxy, alkylthio, alkylamino, (alkyl)2amino, alkylsulfinyl, alkylsulfonyl, arylsulfonyl, carbocyclyl, aryl, or heterocyclyl, wherein R11 is optionally substituted with one or more, the same or different R12; and R12 is halogen, nitro, cyano, hydroxy, trifluoromethoxy, trifluoromethyl, amino, formyl, carboxy, carbamoyl, mercapto, sulfamoyl, methyl, ethyl, methoxy, ethoxy, acetyl, acetoxy, methylamino, ethylamino, dimethylamino, diethylamino, N-methyl-N-ethylamino, acetylamino, N-methylcarbamoyl, N-ethylcarbamoyl, N,N-dimethylcarbamoyl, N,N-diethylcarbamoyl, N-methyl-N-ethylcarbamoyl, methylthio, ethylthio, methylsulfinyl, ethylsulfinyl, mesyl, ethylsulfonyl, methoxycarbonyl, ethoxycarbonyl, N-methylsulfamoyl, N-ethylsulfamoyl, N,N-dimethylsulfamoyl, N,N-diethylsulfamoyl, N-methyl-N-ethylsulfamoyl, carbocyclyl, aryl, or heterocyclyl.