Abstract PO-018: Hypoxia activated prodrugs of DNA-dependent protein kinase inhibitors as radiosensitisers of head and neck squamous cell carcinomas

Abstract

Abstract Inhibition of the repair of radiation-induced DNA double strand breaks offers the potential to sensitize tumors to radiation therapy. DNA-dependent protein kinase (DNA-PK) is a well validated target for radiosensitisation, but inhibition also has the potential to radiosensitize normal tissues within the radiation field, and to target normal tissue functions outside of DNA-PK’s canonical role in DNA repair. Exploiting hypoxia to selectively activate hypoxia-activated prodrugs of DNA-PK inhibitors (DNA-PKi) in tumors may enhance tumor selectivity, with the advantage of sensitizing the most radioresistant (hypoxic) subpopulation, as well as sensitizing adjacent oxic cells through a local bystander effect. We recently conducted a drug discovery campaign, scaffold-hopping from the pan-PIKK inhibitor dactolisib to identify new DNA-PKi. We screened compounds against three signal targets DNA-PK, PI3Kα and mTOR, and determined in vitro sensitization of UT-SCC-54C head and neck squamous cell carcinoma (HNSCC) cells using a growth inhibition endpoint. We identified a highly selective lead DNA-PKi, SN39536, and a corresponding hypoxia-activated prodrug, SN39884. SN39536 is an effective inhibitor of DNA-PK activity, both biochemically and in cells, with substantial selectivity compared to phosphoinositide-3-kinase (PI3K) enzymes and other PI3K-related kinase enzymes. SN39536 provided radiosensitization of DNA-PK proficient HAP1 cells, but was inactive against isogenic cells with knockout of the DNA-PK catalytic subunit (PRKDC), demonstrating on-mechanism activity. SN39536 also provided robust radiosensitization of oxic UT-SCC-54C HNSCC cells using a clonogenic endpoint. Prodrug SN39884 was deactivated as a DNA-PKi and released SN39536 under reductive conditions. Incubation of the prodrug with HCT116 cells overexpressing cytochrome P450 oxidoreductase or UT-SCC-54C cells led to hypoxia-dependent metabolism of the prodrug and efficient release of SN39536. SN39536 effectively sensitized UT-SCC-54C cells to radiation under either aerobic or anoxic conditions, while prodrug SN39884 only sensitized cells under anoxia. SN39536 and its prodrug also demonstrated radiosensitization of hypoxic cells in UT-SCC-54C tumor xenografts using an ex vivo clonogenic survival endpoint. We have identified a new class of potent and selective DNA-PK inhibitors and demonstrated the ability of a hypoxia-activated prodrug to selectively release the inhibitor under hypoxia in vitro and in vivo. Citation Format: Michael P. Hay, Way W. Wong, Benjamin D. Dickson, Gary J. Cheng, Liew P. Liew, Cho R. Hong, William R. Wilson, Stephen M. Jamieson. Hypoxia activated prodrugs of DNA-dependent protein kinase inhibitors as radiosensitisers of head and neck squamous cell carcinomas [abstract]. In: Proceedings of the AACR Virtual Special Conference on Radiation Science and Medicine; 2021 Mar 2-3. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(8_Suppl):Abstract nr PO-018.