BV6 is an antagonist of c-IAP1 and XIAP, members of the inhibitors of apoptosis (IAP) family.

对HCC193细胞处理1 μM BV6 24小时，相对于DMSO处理的细胞，HCC193细胞的生存曲线显著偏移，DER=1.38 (p<0.05)。BV6 (2和5 μM) 显著抑制异位和原位（无病和肌瘤）ESC中的BrdU合成。在使用5 μM BV6处理后，两组中BrdU合成均约减少30%。向HCC193细胞施加1 μM BV6后，治疗1小时即完全耗尽cIAP1水平，而XIAP水平减少则在化合物添加24小时后才出现。同样，5 μM BV6在1小时内完全耗尽H460细胞中的c-IAP1，并在24小时后开始减少XIAP。与此平行的发现是，对于两种细胞系，0.25 μM BV6的小剂量就能减少c-IAP1水平，而在5 μM BV6时，仍可观察到少量的XIAP。从与1 μM BV6孵育12小时后开始，HCC193细胞显著出现切割型caspase-3水平，且这一水平在48小时内随时间增长而持续增加。

小鼠c-IAP-1、c-IAP-2和XIAP表达在植入体的上皮细胞和基质细胞的细胞质中清晰可见，而Survivin主要表达在核内。经过4周的BV6处理后，IAPs表达的强度减弱。免疫组化染色后，角蛋白和波形蛋白呈阳性染色，而钙结合蛋白阴性。采用BV6治疗4周后，病变的总数（4.6对2.8/只小鼠）、平均重量（78.1对32.0 mg/只小鼠）和表面积（44.5对24.6 mm2/只小鼠）显著低于对照组。在子宫内膜腺上皮或基质中，Ki67阳性细胞的百分比在BV6治疗后降低。

BV6 is prepared in DMSO and stored, and then diluted with appropriate medium before use. H460 and HCC193 cell lines are cultured in RPMI-1640 supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. Cell viability is measured using the CellTiter 96 Aqueous Non-Radioactive Cell Proliferation Assay kit. 5000 cells/well are seeded into 96-well plates in triplicate. Following adhesion of cells to the wells, increasing concentrations of BV6 are added into different wells. Control groups are exposed to the same concentration of DMSO. The final concentrations of 333 μg/mL MTS and 25 μM PMS are added to each well 24 hours later. After two hours incubation at 37°C in humidified 5% CO2, plates are read at the absorbance of 490 nm on a microplate reader. Relative cell viability of an individual sample is calculated by normalizing their absorbance to that of the corresponding control. IC50 values are calculated using Prism 5.01. For the TNFα neutralizing antibody assay, cells are exposed to 1 and 5 μM BV6 with or without 10 μg/mL Infliximab and the assay is performed 24 hours later. Plates are read at the absorbance of 490 nm on a microplate reader.

BV6 is prepared in DMSO and diluted with saline or PBS. Female mice (6 weeks of age, BALB/c) are used. All 24 mice are ovariectomized through a 1 cm longitudinal skin incision then injected s.c. with estradiol valerate (0.5 μg/mouse/week) once per week for 6 weeks until the experimental endometriosis induction. Two weeks after ovariectomy, the uteri of an additional eight donor mice (n=8) are removed en bloc after euthanasia and cleaned of excess tissue in sterile saline. Each uterus is cut to include the uterine horns in each half with a linear incision longitudinally and minced (0.5 mm in diameter) with dissecting scissors. The ovariectomized recipient mice (n=16) are anesthetized using pentobarbital sodium. A 0.5 cm subabdominal midline incision is made. Each recipient receives half of the donor uterus (1:2 donor uterus to host ratio) minced and added to 500 μl saline, and injected into the peritoneal cavity, and the peritoneum is sutured. Injected uterine tissue weighed ~50 mg per mouse. For the next 4 weeks, recipient mice are treated with a single i.p. injection of BV6 (n=8; 10 mg/kg) or vehicle (n=8; 1% DMSO) twice weekly.