Rucaparib Phosphate (PF-01367338 phosphate) is an inhibitor of PARP (Ki: 1.4 nM for PARP1) that is used in clinical therapy to sensitize cancer cells to chemotherapy.

PARP:1.4 nM (Ki, cell free)

AG14447（Rucaparib的磷酸盐）在酶测定中是高效的PARP抑制剂（Ki：1.4 nmol/L）[1]。在渗透的D283Med细胞中，Rucaparib (AG-014699) 以0.1, 0.4 和 1μ的浓度分别抑制了81.1%，96.8% 和97.1% 的PARP-1活性[2]。AG014699（≤10 μM）对带有突变BRCA1/2或XRCC3的细胞以及表观遗传学沉默BRCA1的UACC3199细胞具有细胞毒性，但对没有BRCA1/2或XRCC3突变或BRCA2突变为杂合子的细胞则无影响[3]。

当以10mg/kg Rucaparib与temozolomide联合给药时，观察到治疗后第4至13天体重减轻，体重最低点范围从起始体重的83%至96%。以1mg/kg剂量，Rucaparib也显著增加了由temozolomide引起的体重减轻[1]。以1mg/kg、每日五次给药，AG-014699单独使用未引起任何明显毒性或对肿瘤生长产生影响，与仅用载体对照组比较。AG-014699与temozolomide的联合给药也导致所有小鼠的肿瘤完全消退，其中五分之三的小鼠实验期间肿瘤持续消退。MMR缺陷的D283Med异种移植物生长非常迅速，对单独使用temozolomide的响应极低（TGD仅为2天），在任何小鼠中未观察到肿瘤退缩[2]。

We measured inhibition of human full-length recombinant PARP-1 by [32P]NAD+ incorporation as described previously. The [32P]ADP-ribose incorporated into acid insoluble material was quantified using a PhosphorImager. Kis were calculated by nonlinear regression analysis [1].

Inhibition of PARP activity in 5000 exponentially growing D283Med cells was measured following treatment with a range of AG-014699 concentrations (0–1?μ), in comparison with DMSO-only controls. Maximally stimulated PARP activity was measured in replicate samples (n?3) of permeabilised cells by immunological detection of the amount of poly(ADP-ribose) (PAR) formed, using 10H anti-PAR antibody, during a 6-min incubation with NAD+ and oligonucleotide (substrate and activator) by reference to a PAR standard curve using a GCLP-validated assay described previously [2].

One or four daily doses of PARP inhibitor AG-014699 (1?mg/kg intraperitoneally (i.p.)) were given to CD-1 nude mice bearing established D283Med xenografts. At 0.5, 2, 6 and 24?h after the initial or fourth daily dose of AG-014699, three animals per time point were bled by cardiac puncture under general anaesthesia, and then killed. Plasma was separated from the blood samples using standard methods and stored at ?80°C. The brains and tumours were removed, snap frozen in liquid nitrogen and stored at ?80°C before analysis. Blood, tumour and brain tissue were removed from three untreated control animals and processed in the same way [2].