Abstract 7166: Pharmacokinetic analysis of omomyc therapeutic protein in preclinical and human tumor tissue using targeted mass spectrometry

Abstract

Abstract MYC is a pleiotropic transcription factor consisting in three paralogs which coordinate gene expression and regulates cell proliferation, cell differentiation, cell cycle, metabolism and apoptosis. The 3 paralogs (c-, N-, and L-MYC) have similar biological functions but show distinct tissular and timing expression patterns. To function, MYC must heterodimerize with its partner MAX and bind DNA via its basic, helix-loop-helix, leucine zipper domain (B-HLH-LZ). It has been estimated that MYC is aberrantly expressed in up to 70% of human cancers and hence MYC has been termed a “most wanted target for cancer treatment”. However, due to its nuclear localization, its intrinsic disorder and lack of an enzymatically active site, development of clinical grade potent & selective small molecules targeting MYC remains an obstacle. Omomyc is a 91-amino-acids-long polypeptide that derives from the B-HLH-LZ domain of human MYC, containing four amino acid substitutions. Omomyc is capable of forming homodimers and heterodimers with MYC and with MAX and such dimers blunt MYC activity by preventing its access to target DNA sequences (E-boxes) and replacing it by inactive complexes. Previous studies showed the cell-penetrating properties of recombinant Omomyc and its subsequent nuclear localization, where it displaces MYC from its genomic locations, causing a specific shutdown of MYC transcriptional signature and effectively stopping cell cycle progression. Further in vivo preclinical experiments showed that Omomyc blocks MYC-driven transcription and proliferation, induces apoptosis and recruitment of intratumoral immune infiltrates, overall significantly abrogating tumor progression and reducing tumor grading. A targeted LC-MS/MS assay for the absolute quantification of 4 Omomyc proteotypic peptides was developed to assess the structural integrity of Omomyc and quantify functional Omomyc in tumor tissue. LLOQ and linear range of the panel was determined by spiking in dilution series of recombinant Omomyc protein into preclinical tumor tissue lysates. Finally, in the context of Omomyc clinical trial (NCT04808362), we used the panel to for quantification of functional Omomyc in FFPE tumor biopsies from 18 patients who received i.v. injection of Omomyc therapeutic. Data show that 2 h after i.v. administration, higher concentrations of the drug are reached in the tumors compared with serum and persist there with at least one order of magnitude higher concentration than in serum 72 h after administration in preclinical models. Functional Omomyc protein could be quantified in all of the 18 clinical tumor biopsies from treated patients showing that characterized LC-MS/MS proteomic assay can be used in the normal routine of a clinical trial for accurate quantification of therapeutic miniproteins. Citation Format: Marie-Eve Beaulieu, Sandra Martinez-Martin, Maik Mueller, Yuehan Feng, Jakob Vowinckel, Antoine Amaury Lachaud, Laura Soucek. Pharmacokinetic analysis of omomyc therapeutic protein in preclinical and human tumor tissue using targeted mass spectrometry [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7166.