BPTES

BPTES在IDH1突变的D54细胞中优先减缓细胞生长。BPTES还抑制谷氨酰胺酶活性，降低谷氨酸和α-KG水平，并增加糖酵解中间体。BPTES (10 μM)可抑制LAP/MYC肿瘤来源的mHCC 3-4细胞的生长。BPTES还通过阻断DNA复制抑制myc依赖的P493细胞的生长，导致细胞死亡和分裂。BPTES (10 μM) 在 A549 和 EKVX 细胞系中与 10 μM 5-FU 显示出明显的协同抗癌作用，不仅在 EKVX 和 A549 中而且在大多数 NSCLC 细胞系中都会导致生长减少反应。

BPTES (12.5 mg/kg, i.p.) 在LAP/MYC小鼠中，延长存活期，而对MYC，GLS，或GLS2水平没有显著作用。BPTES (200 μg/mouse, i.p.) 在负荷P493肿瘤异种移植物的小鼠中，也会抑制肿瘤细胞生长。BPTES -NP (BPTES 纳米颗粒，100 μL 纳米颗粒中含有 1.2 mg BPTES，静脉注射) 显著减弱患者来源的胰腺原位肿瘤模型中的肿瘤生长。

BPTES treatment of mouse HCC model.
MYC expression was induced in LAP/MYC mice by doxycycline removal on the day of birth. LAP/MYC mice were randomly assigned to the BPTES or vehicle treatment group. Intraperitoneal injections (12.5 mg/kg body weight every 3 days) of BPTES or vehicle control (10% DMSO in 200 μl of PBS every 3 days) were initiated 3 weeks after birth.

BPTES treatment of tumor xenografts.
P493 cells (2 × 10⁷) were injected subcutaneously into the flank of athymic nude mice. When the tumor volumes reached approximately 100 mm3, intraperitoneal 0.2 ml injections of BPTES (200 μg) or vehicle control (10% DMSO in PBS) were initiated and carried out every 3 days for 10 days. The tumor volumes were measured using digital calipers every 3 days and calculated using the following formula: length (mm) × width (mm) × width (mm) × 0.52.

Hematology.
Blood samples from BALB/c mice treated for 2 weeks with BPTES (12.5 mg/kg body weight every 3 days) or vehicle (10% DMSO in 200 μl of PBS every 3 days) were analyzed using the Hemavet 950FS hematology system using the mouse key.

Toxicity study.
After a 14-day quarantine period, BALB/c mice were assigned at random to 2 groups. The study included a treatment group (12.5 mg/kg body weight every 3 days) and a control group (10% DMSO in 200 μl of PBS every 3 days). Mice were treated for 10 days (4 injections on days 1, 4, 7, and 10). Observations were made twice daily for clinical signs of pharmacologic and toxicologic effects of the BPTES. Histopathologic evaluation of brain, heart, skeletal muscle, lung, and kidney was done by a pathologist without any animal group information.

Primary human T cell proliferative analysis.
Primary human T lymphocytes were cultured in RPMI 1640 supplemented with 10% FBS, 10 mM HEPES, 2 mM l-glutamine, 10 U/ml penicillin G, and 100 μg/ml streptomycin. T cells were activated with 4.5 μm microbeads containing immobilized anti-human CD3 and anti-human CD28 at a ratio of 3 beads to 1 cell. After 72 hours of stimulation in the presence of either DMSO or BPTES (10 μM), T cells were counted by flow cytometry using CountBright Beads, Via-Probe, and mAbs to human CD4/CD8. Cell volume was assayed using a Multisizer III particle counter.

Indirect coating of tissue culture surfaces with Abs for T cell stimulation.
Twelve-well tissue culture plates were treated with 10 μg/ml goat anti-mouse IgG overnight at 4°C. The plates were then rinsed 3 times with PBS, and a blocking buffer containing 5% BSA in PBS was applied for 1 hour. After a series of washes, the plates were incubated with 5 μg/ml of Okt3 for 2 hours. Following a final series of washes, T cells were seeded at 1 × 10⁶ cells/well and supplemented with soluble 9.3. Additionally, T cells were treated with either DMSO or BPTES (10 μM). After 72 hours, T cells were counted by flow cytometry using CountBright Beads and Viaprobe. Cell volume was assayed using a Multisizer III particle counter.

https://pmc.ncbi.nlm.nih.gov/articles/PMC4497742/