Abstract C009: Clinical pharmacokinetic and pharmacodynamic analyses support biological effectivity of the PARP inhibitor olaparib as radiosensitizer at low doses
Authors: Haan Rosemarie de , Pluim Dick , Verwijs Manon , Sonke Gabe , Heuvel Michel van den , Triest Baukelien van , Verheij Marcel , Vens Conchita 
Published: March 2020

Abstract
Introduction: PARP inhibitors are promising radiosensitizers. Currently identified recommended phase II doses of PARP inhibitors in combination with radiotherapy are 10-fold lower than the EMA and FDA approved dose as single agent. The purpose of this study is to investigate the biologically effective dose range of olaparib for its use as radiosensitizer by assessing pharmacokinetics (PK) and pharmacodynamics (PD). Both the inhibition of pre-treatment PAR levels and the inhibition of radiation induced PARylation therefore have to be determined. Methods: We included PK and PD samples of all NSCLC and breast cancer patients treated at olaparib doses up to 50mg bi-daily (BID) in two phase 1 trials combining radical radiotherapy with olaparib. Plasma samples were collected for PK analysis (ranging from steady state pre-dose olaparib until +12/24 hrs after intake) and peripheral blood mononuclear cells (PBMCs) for PD analysis (pre-treatment, +3 hrs and +12/24 hrs after olaparib intake). Breast cancer patients with the primary tumor in situ had biopsies taken pre-treatment and during olaparib treatment (+3 hrs after intake) before the first fraction of radiotherapy. Olaparib concentrations were determined by HPLC-MS/MS. PAR levels were determined by commercially available ELISA, following both the NCI protocol and REP assay (de Haan et al, 2017) that includes ex vivo irradiation of intact cells to activate PARylation by PARP. PAR levels determined by the REP assay were used to calculate PAR level inhibition during treatment. Results: PK/PD data from blood samples were available from 28 NSCLC patients and 7 breast cancer patients treated with radical radiotherapy and olaparib (17 at 25mg once daily (QD), 14 at 25mg BID and 4 at 50mg BID). Repeat biopsies were available from six out of seven breast cancer patients. The plasma olaparib concentrations increased with the dose levels and showed an expected wide intra-patient variation. In tumors the olaparib concentration varied between 178 and 1441 ng/g with a median tumor to plasma ratio of 0.47. PD analyses demonstrate a significant reduction in PAR levels in PBMCs during olaparib treatment (&gt;95% at +3 hrs after olaparib intake for all dose levels, &gt;90% at +12 hrs in the 25mg BID dose level and 66-99% at +24 hrs in the 25mg QD dose level; all p&lt;0.0001). In pre-treatment samples, ex vivo radiation induced PAR levels by 66-fold (range 23 to 174). This radiation induced PARylation was abolished during olaparib treatment. Compared to corresponding pre-treatment PBMC samples, PAR levels in pre-treatment tumor biopsy samples were higher in all but one patient (median 7-fold higher, range 0.6-60), and increased by only 1.5-fold after ex vivo radiation. Also in tumors, olaparib abolished radiation induced PARylation. PAR levels were reduced by 89% (range 83-99.7%, p&lt;0.0001). Conclusion: Olaparib doses as low as 25mg once and twice daily inhibit PARP activation by irradiation and reduce PAR levels &gt;95% in PBMCs and &gt;83% in tumors, thereby showing biological effectivity. The tumor olaparib concentrations determined in this study were all within a range that has been shown to radiosensitize in preclinical models. Together this supports further development of PARP inhibitors as radiosensitizer at low doses.
Citation Format: Rosemarie de Haan, Dick Pluim, Manon Verwijs, Gabe Sonke, Michel van den Heuvel, Baukelien van Triest, Marcel Verheij, Conchita Vens. Clinical pharmacokinetic and pharmacodynamic analyses support biological effectivity of the PARP inhibitor olaparib as radiosensitizer at low doses [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr C009. doi:10.1158/1535-7163.TARG-19-C009