生物活性

PX-478 2HCl is an inhibitor of HIF-1a with an IC50 of 20-30μM enhances radiosensitivity of prostate carcinoma cells. PX-478 2HCl enhances the radiosensitivity of prostate carcinoma cells irradiated under normoxia. In addition to the inhibition of HIF-1a, treatment with PX-478 2HCl resulted in cell type-dependent alterations in cell cycle distribution and prolonged gH2AX expression in the irradiated cells. In PC3 cells, PX-478 2HCl significantly reduced the percentage of cells in G1 with an accumulation of cells in S/G2M. PX-478 is an orally active selectiveinhibitor of HIF-1α translation that suppresses constitutive andhypoxia-induced HIF-1 in cancer cell lines and shows antitumor activity againstseveral aggressive human tumor xenografts.

PX-478 inhibits HIF-1α with an IC50 of 5 and 7μM in MCF-7 human breast and HT-29 humancolon cancer cells, respectively. PX-478inhibits HIF transactivating activity in MCF-7 and HT-29 cells with an IC50 of 7-9μM. PX-478inhibits hypoxia-induced VEGFR formation by MCF-7 and HT-29 colon cancer cells (IC507-9μM). [1]

PX-478inhibits the activity of a HIF-1–dependent GFP reporter construct transfectedinto C6 cells with an IC50 value of 49.2 μM.[2]

PX-478inhibits the growth of Panc-1 and BxPC-3 cells with IC50 around 40μM.[3]

Cellabilities of PX-478[4]

体外研究

体内研究

激酶实验

细胞实验

Cells[6]

BxPC-3, Panc-1, SU.86.86, CF-PAC-1, andMiaPaCa-2 pancreatic cancer cells were obtained. MiaPaCa-2 cells were stably transfectedwith short hairpin RNA (shRNA) specific to HIF-1α. The cells were grown inhumidified 95% air, 5% CO₂ at 37°C in DMEM supplemented with 10%fetal bovine serum. All cell lines were tested to be mycoplasma free using a PCRenzyme-linked immunosorbent assay kit.

Clonogenicsurvival assays

Panc-1 and BxPC-3 cells were grown to 70%confluence and incubated at 21% O₂ (normoxia) or 1% O₂ (hypoxia,via chamber) for 24 hours. Cells were treated with 25μmol/L PX-478 for 24 hoursand irradiated with a ¹³⁷Cs source (5.8 Gy/min), with atmosphericconditions maintained. Cells were assayed for colony formation by replating atspecified numbers into 6-well plates in drugfree medium. The cells wereimmediately plated after irradiation, maintained for 12 days in normoxia, and stainedwith 0.5% crystal violet in absolute ethanol. Colonies with >50 cells werecounted. Clonogenic survival curves were constructed from at least threeindependent experiments, and the radiation sensitizer enhancement ratio at 0.2surviving fraction was calculated.

动物实验

Animal experiments[3]

Panc-1 cells were re-suspended in PBS at 10⁷/mL.Each of the 40 nude mice was injected with 100μL of the suspensionsubcutaneously. Arsenic trioxide (ATO) was injected at 6 mg/kgintraperitoneally 3 times a week. PX-478 was delivered via p.o. gavage at 30mg/kg every other day. Tumors were harvested at week 5. Tumor volume wascalculated as the product of all three dimensions. Immunohistochemistry for HIF-1a,FOXO1, SESN3, Cleaved Caspasse-3, and Ki67 of mouse tumor tissue was performed withcorresponding antibodies and DAB substrate kit according to instructions. Theexpression levels of each protein were mounted as percentage of positive cells.The mouse frozen sections were stained with 10μM of Dihydroethidium for 30 minat room temperature in the dark. Cell number was calculated by DAPI nuclearstaining. All pictures were taken at 200-fold magnification. DHE intensity was analyzedusing Image J software. All the animal experiments were performed in accordancewith the ARRIVE guidelines for the care and usage.

不同实验动物依据体表面积的等效剂量转换表（数据来源于FDA指南）

例如，已知某工具药用于小鼠的剂量为88 mg/kg , 则用于大鼠的剂量换算方法：将88 mg/kg 乘以小鼠的Km系数（3），再除以大鼠的Km系数（6），得到该药物用于大鼠的等效剂量44 mg/kg。

参考文献

[1] Welsh SJ WR, Kirpatrick DL, Powis G. PX-478, a potent inhibitor of hypoxia-inducible factor-1 (HIF-1) and antitumor agent. Eur J Cancer. 2002;38(Suppl 7):S90.
[2] Schwartz DL, Powis G, Thitai-Kumar A, et al. The selective hypoxia inducible factor-1 inhibitor PX-478 provides in vivo radiosensitization through tumor stromal effects. Mol Cancer Ther. 2009;8(4):947-958.
[more]

体内溶解度

Clinical Trial Information ( data from http://clinicaltrials.gov )

注：以上所有数据均来自公开文献，并不保证对所有实验均有效，数据仅供参考。

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