Degrasyn (WP1130) (WP1130), a specific deubiquitinase (DUB: USP5, UCH-L1, USP9x, USP14, and UCH37) inhibitor, also inhibits Bcr/Abl, which is a JAK2 transducer (without affecting 20S proteasome) and activator of transcription (STAT).

BCR-ABL:1.8 μM

除了快速下调Bcr/Abl而不影响Bcr或c-Abl之外，WP1130还调节Jak2和c-Myc的稳定性，但不影响其他激酶（HER1, HER2, c-Kit, FAK, ERK1, ERK2, Akt, Btk, Src及Src相关激酶）或转录因子（野生型p53, STAT1, STAT3, STAT5, c-Jun, NF-κB及Max）。与adaphostin和Trisenox不同，WP1130在60分钟内就能引起Bcr/Abl的下调。WP1130诱导髓系和淋巴系肿瘤细胞凋亡的效率更高，IC50约为0.5-2.5 μM。与之相比，对正常CD34+造血前体细胞、皮肤成纤维细胞或内皮细胞的IC50约为5-10 μM。WP1130（5 μM）特异性和快速地下调野生型及T315I突变型Bcr/Abl蛋白，而不影响bcr/abl基因表达或介入蛋白酶体降解途径，在慢性髓性白血病（CML）细胞中伴随着凋亡的诱导。WP1130在减少白血病细胞集落形成方面比对正常祖细胞更为有效，并且能有效针对携带T315I突变的原发性白血病细胞。WP1130诱导MM-1多发性骨髓瘤和其他肿瘤细胞系中c-Myc蛋白的快速蛋白酶体依赖性降解，与肿瘤生长抑制相关。与AG490不同，WP1130作为部分选择性脱泛素化酶（DUB）抑制剂，能引起快速而显著的polyubiquitinated (K48/K63-linked)蛋白质积累于近核聚集体，而不影响蛋白酶体活性。WP1130（5 μM）直接抑制USP9x, USP5, USP14, UCH-L1及UCH37的DUB活性，但不影响UCH-L3，导致抗凋亡蛋白下调及促凋亡蛋白上调，如MCL-1和p53。

WP1130的施用抑制了裸鼠体内移植的K562肿瘤以及野生型Bcr/Abl和T315I突变型Bcr/Abl表达的BaF/3细胞的生长。[1] 与c-Myc的下调一致，WP1130在裸鼠体内建立的A375黑色素瘤肿瘤中展示了强大的抑制活性。[2]

To determine binding kinetic constants, liver or kidney plasma membranes are incubated with increasing concentrations of [3H]-AVP with or without excess (1 μM) unlabelled AVP to obtain a saturation curve. To investigate whether mozavaptan interacts competitively or noncompetitively, the saturation binding of [3H]-AVP is examined in the absence and presence of mozavaptan at concentrations of 0.3 μM and 1 μM in liver membranes and 3 nM, and 10 nM in kidney membranes. Data on the saturation curve are plotted according to the method of Scatchard and fitted by a regression analysis[1].

Cells are treated with increasing concentrations of WP1130 (0.08-10 μM) in 96-well plates. Plates are incubated at 37 °C for 72 hours, after which 20 μL of MTT reagent is added, and the plates are incubated at 37 °C for another 2 hours. Cells are lysed with 100 μL lysis buffer (20% sodium dodecyl sulfate [SDS] in 50% N, N-dimethylformamide adjusted to pH 4.7 with 80% acetic acid and 1 M hydrochloric acid; final concentration of acetic acid is 2.5% and hydrochloric acid is 2.5%) and incubated for 6 hours. The optical density of each sample at 570 nm is determined with a SPECTRA MAX M2 plate reader.(Only for Reference)