MI-2 (MALT1 inhibitor) is an irreversible MALT1 inhibitor.

MALT1:5.84 μM

每天腹腔注射25 mg/kg MI-2高效抑制TMD8 和 HBL-1 ABC-DLBCL异种移植物的生长,而对OCI-Ly1肿瘤的生长无作用.

MI-2具有良好的细胞渗透性,抑制ABC-DLBCL细胞系中MALT1功能。MI-2对MALT1依赖性细胞系产生选择性生长抑制作用,在HBL-1,TMD8,OCI-Ly3和OCI-Ly10细胞中GI50分别为0.2,0.5,0.4和0.4 μM,而ABC-DLBCL MALT1不依赖的细胞系U2932 和 HLY-1以及两种GCB-DLBCL细胞系对MI-2都具有抗性。

High-Throughput Screening for MALT1 Proteolytic Activity Inhibitors: Ac-LRSR-AMC is used as substrate and reactions are measured with excitation/emission wavelengths of 360/465 nm. Two time points are measured for each reaction to eliminate false positives due to compound autofluorescence. The final percent inhibition is calculated with the formula {[fluorescencetest compound (T2-T1)-fluorescenceneg ctrl (T2-T1)]/[fluorescencepos ctrl (T2-T1)-fluorescenceneg ctrl (T2-T1)]} × 100, where Z-VRPR-FMK (300 nM) is used as positive control and buffer only as negative control. The positive hits are validated in concentration-response experiments within a dose range of 0.122–62.5 μM to determine IC50 of the compounds. Activity is validated using recombinant full-length wild-type MALT1.

Cell proliferation is determined by ATP quantification using a luminescent method and trypan blue dye exclusion. Standard curves for each cell line are calculated by plotting the cell number (determined using trypan blue) against their luminescence values, and cell number is calculated accordingly. Cell viability in drug-treated cells is normalized to their respective controls (fractional viability), and results are given as 1-fractional viability. CompuSyn software is used to determine GI25 and GI50 values.(Only for Reference)