Abstract LB-060: A very long-acting poly(ADP-ribose) polymerase inhibitor

Abstract

Abstract Purpose: The purpose of this work was to prepare and characterize the anti-tumor properties of a prodrug for a very long-acting of poly(ADP-ribose) polymerase (PARP) inhibitor. Background: Four PARP inhibitors (PARPi) have been approved for QD oral use in treatment of human cancers. It is believed that PARP requires continuous inhibition for optimal anti-tumor effects, but, as with any drug with a short t1/2, daily administered PARPi exhibit high Cmax values and peak-to-trough excursions. We speculated that the prolonged exposure and lower Cmax and Cmax/Cmin of a long-acting PARPi might provide a more effective, less toxic therapeutic. In choosing which PARPi to target, a major consideration was whether the carrier has capacity to deliver sufficient levels of the drug over a long dosing interval. Talazoparib (TLZ) was chosen for this study because it is the most potent of the PARP inhibitors, requiring only 1 mg/day in adults compared to hundreds of mg/day for other PARPi's. Experimental procedures: We prepared a long-acting prodrug of TLZ by attaching it to a PEG40kDa carrier by a β-eliminative releasable linker. The chemistry was achieved by a novel alkylation of TLZ at the poorly acidic 2-NH of the phthalazinone moiety with an O-azidoalkyl-N-alkyl-N-chloromethyl carbamate, followed by coupling to PEG-cyclooctyne. Daily PO doses of TLZ or a single IP injection of the PEG-TLZ conjugate were administered to xenografts in mice possessing defects in homologous recombination - either a PALB2 mutation in the KT-10 Wilms tumor, or a BRCA1-deficient MX-1 triple-negative breast cancer. New data: PEG~TLZ was highly effective in treating both KT-10 and MX-1 xenografts. Although the t1/2 of TLZ in the mouse is only ~3 hr, tumor growth in animals treated with PEG~TLZ was suppressed for about one month. The EFS T/C values - the ratio of the median time to event between treated and control groups - of single injections of ~5 mg TLZ/kg as PEG~TLZ in either tumor was more than 4, indicating the drug is a highly active agent at low doses. The amount of TLZ in a single efficacious dose of the PEG~TLZ conjugate was equivalent to the same amount of free TLZ administered in divided daily doses for 4 or more weeks. Although we did not investigate scheduling, dosing PEG~TLZ once every 3 to 4 weeks should be sufficient to suppress tumor growth for extended periods. Conclusion: We developed a novel method of conjugating linkers to the 2N of the phthalazinone moiety of PARPi. We prepared a cleavable PEG~TLZ that releases TLZ with a t1/2 of 160 hr at pH 7.4. In mouse xenografts of tumors with defective HR, single non-toxic doses of PEG~TLZ suppresses tumor growth for ~1 month, and are equi-effective to QD administration of TLZ over that period. We posit that the long lasting effect is due to the long t1/2 of the prodrug, increased sensitivity of the tumor upon continued exposure to TLZ, tumor accumulation of the 15 nm nanomolecule, and counteracting drug resistance by efflux pumps. Citation Format: Shaun D. Fontaine, Gary W. Ashley, Peter J. Houghton, Raushan Kurmasheva, Morgan Diolati, Alan Ashworth, Daniel V. Santi. A very long-acting poly(ADP-ribose) polymerase inhibitor [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr LB-060.