CRT0066101 dihydrochloride is an effective and selective PKD inhibitor (IC50s: 1, 2.5 and 2 nM for PKD1, 2, and 3).

PKD3:2 nM, PKD2:2.5 nM, PKD1:1 nM

CRT0066101专门阻断PKD1/2活性，不抑制多种癌细胞类型（包括A549和MiaPaCa-2）中的PKCα/PKCβ/PKCε活性。CRT0066101明显抑制Panc-1细胞增殖（IC50: 1 μM），治疗导致Panc-1细胞凋亡诱导增加6-10倍。此外，CRT0066101显著降低Panc-1、Colo357、MiaPaCa-2和AsPC-1细胞的增殖，但对Capan-2细胞的影响较轻。CRT0066101（5 μM）同时阻断Panc-1和Panc-28细胞中基础和NT诱导的pS916-PKD1/2（活化的PKD1/2）。CRT0066101减少PKD依赖的NF-κB激活及在Panc-1细胞中的NF-κB依赖基因表达。

口服给药CRT0066101（每日80 mg/kg）28天可显著抑制Panc-1皮下异种移植模型中的PaCa生长。在治疗后的肿瘤组织中，CRT0066101在口服后2小时内达到峰值肿瘤浓度（12 μM），显著抑制激活的PKD1/2表达。此外，CRT0066101口服给药（每日80 mg/kg）在Panc-1原位模型中持续21天，强效阻断体内肿瘤生长。CRT0066101显著降低Ki-67+增殖指数，增加TUNEL+凋亡细胞，并抑制包括cyclin D1、survivin和cIAP-1在内的NF-κB依赖性蛋白的表达。在口服给药后6小时，可检测到CRT0066101的最佳治疗浓度（8 μM）。

Nineteen days after implantation of Panc-1 cells, tumor areas are, on average 0.3 cm2 and are randomized into the following groups (n=8 mice per group): (a) vehicle (control) 5% dextrose administered by oral gavage once daily and (b) 80 mg/kg CRT0066101 dissolved in 5% dextrose administered by oral gavage once daily. Tumors are measured in 2 dimensions every 2 to 3 days by calipers and area is calculated by multiplying the length by width. Therapy is given until tumors reached their designated size limits (1.44 cm2) or until day 24 in CRT0066101 treated group. Final tumor areas are compared among groups using a Student's t-test and Fisher's exact test.