PJ34 HCl is the hydrochloride salt of PJ34, which is a PARP inhibitor with EC50 of 20 nM and is equally potent to PARP1/2.

PARP:20 nM (EC50)

PJ34通过抑制PARP酶活性，其IC50值为110±1.9 nM。通过LDH测定法评估PJ34与其他PARP抑制剂在PC12细胞中的神经保护性能，比较结果表明，PJ34在从10-7到10-5 M范围内的浓度下，显著并且依赖浓度地减少细胞死亡[1]。

为了与其他PARP抑制剂进行效能及功效的比较，评估PJ34的剂量分别为3.2和10 mg/kg。3.2 mg/kg的剂量显著降低了皮质损伤33%，而10 mg/kg的剂量则显示出相反效果（降低了17%）[1]。25 mg/kg的PJ34能够将缺血动物的TNF-α mRNA水平降低70%，且这些数值与假手术或未处理动物无差异。PJ34对缺血小鼠的治疗降低了E-selectin mRNA的水平81%，ICAM-1 mRNA的水平降低了54%，与仅接受载体处理的缺血小鼠相比[2]。

To assess the PARP-1 or PARP-2 inhibitory activity of FR247304, 3-AB, and PJ34, PARP activity is evaluated with minor modifications. PARP enzyme assay is carried out in a final volume of 100 μL consisting of 50 mM Tris-HCl (pH 8.0), 25 mM MgCl2, 1 mM dithiothreitol, 10 μg activated salmon sperm DNA, 0.1 μCi of [adenylate-32P]NAD, 0.2 units of recombinant human PARP for PARP-1 assay or 0.1 units of recombinant mouse PARP-2 for PARP-2 assay, and various concentrations of FR261529 or 3-AB. The reaction mixture is incubated at room temperature (23°C) for 15 min, and the reaction is terminated by adding 200 μL of ice-cold 20% trichloroacetic acid (TCA) and incubated at 4°C for 10 min. The precipitate is transferred onto GF/B filter and washed three times with 10% TCA solution and 70% ethanol. After the filter is dried, the radioactivity is determined by liquid scintillation counting.

PJ34 is dissolved in 100% DMSO at 10 mM and then diluted in DMEM without serum[1]. PC12 cell cultured are grown in Dulbecco's modified Eagle's medium supplemented with 5% (v/v) fetal calf serum, 5% (v/v) horse serum, and a 1% (v/v) penicillin-streptomycin antibiotics mixture. Cells are grown in an atmosphere of 95% air and 5% CO2 at 37°C. For all experiment, cells are seeded at a density of 4×104 cells/well in 96-well culture plates and allowed to attach overnight. For assessment of cell viability, hydrogen peroxide-induced cytotoxicity is quantified by a standard measurement of LDH release with the use of the LDH assay kit. Briefly, 6 h after hydrogen peroxide exposure, 20 μL of medium of each well is collected, and the solution prepared from LDH assay kit is added. After incubation at room temperature for 30 min, the reaction is stopped by addition of 1 N HCl, and absorbance is measured at 450 nm using a microplate reader.