7e661def6e86c94406c0d06a572f747a pmc_9913332.pdf 82c9ac8f63020d1095bd8d457eadde8d53e622e6 pmc_9913332.pdf ccf75e1d4b119327710fe3e09d00aba8fb049fb51d4b0700f73d1e33281ae3d0 pmc_9913332.pdf Title: Pharmacokinetic Analysis of Omomyc Shows Lasting Structural Integrity and Long Terminal Half-Life in Tumor Tissue Subject: MYC is an oncoprotein causally involved in the majority of human cancers and a most wanted target for cancer treatment. Omomyc is the best-characterized MYC dominant negative to date. In the last years, it has been developed into a therapeutic miniprotein for solid tumor treatment and recently reached clinical stage. However, since the in vivo stability of therapeutic proteins, especially within the tumor vicinity, can be affected by proteolytic degradation, the perception of Omomyc as a valid therapeutic agent has been often questioned. In this study, we used a mass spectrometry approach to evaluate the stability of Omomyc in tumor biopsies from murine xenografts following its intravenous administration. Our data strongly support that the integrity of the functional domains of Omomyc (DNA binding and dimerization region) remains preserved in the tumor tissue for at least 72 hours following administration and that the protein shows superior pharmacokinetics in the tumor compartment compared with blood serum. Keywords: protein therapeutics; MYC; Omomyc; mass spectrometry; LC-PRM Author: Marie-Eve Beaulieu, Sandra Martínez-Martín, Jastrinjan Kaur, Virginia Castillo Cano, Daniel Massó-Vallés, Laia Foradada Felip, Sergio López-Estévez, Erika Serrano del Pozo, Hugo Thabussot and Laura Soucek Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Sun Jan 29 09:07:13 2023 CET ModDate: Sun Jan 29 09:30:18 2023 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 10 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 934063 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- XWOCXD+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 CIQLLF+VnURWPalladioL Type 1 Custom yes yes no 15 0 GLRDNN+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 CBIWKN+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 LQOZWS+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 LCJEVJ+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 47 0 VYVZGZ+CMSY10 Type 1 Builtin yes yes no 72 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 76 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 91 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 95 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 99 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 103 0 BCDEEE+PalatinoLinotype-Bold TrueType WinAnsi yes yes no 120 0 BCDFEE+PalatinoLinotype-Roman TrueType WinAnsi yes yes no 124 0 BCDGEE+PalatinoLinotype-Italic TrueType WinAnsi yes yes no 128 0 BCDIEE+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 132 0 SymbolMT CID TrueType Identity-H yes no yes 139 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2023-12-23 02:29:37 CET RepresentationInformation: pmc_9913332.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2023-04-22 02:23:51 CEST Size: 934063 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 256 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Production of Serum and Tumor Samples Destination: subsection.2.1 Item: Title: Sample Preparation for Mass Spectrometry Destination: subsection.2.2 Item: Title: Shotgun LC MS/MS for Spectral Library Generation Destination: subsection.2.3 Item: Title: LC-PRM Mass Spectrometry Acquisition and Data Analysis Destination: subsection.2.4 Item: Title: Statistical Analysis Destination: subsection.2.5 Item: Title: Results Destination: section.3 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: section.5 Info: Title: Pharmacokinetic Analysis of Omomyc Shows Lasting Structural Integrity and Long Terminal Half-Life in Tumor Tissue Author: Marie-Eve Beaulieu, Sandra Martínez-Martín, Jastrinjan Kaur, Virginia Castillo Cano, Daniel Massó-Vallés, Laia Foradada Felip, Sergio López-Estévez, Erika Serrano del Pozo, Hugo Thabussot and Laura Soucek Subject: MYC is an oncoprotein causally involved in the majority of human cancers and a most wanted target for cancer treatment. Omomyc is the best-characterized MYC dominant negative to date. In the last years, it has been developed into a therapeutic miniprotein for solid tumor treatment and recently reached clinical stage. However, since the in vivo stability of therapeutic proteins, especially within the tumor vicinity, can be affected by proteolytic degradation, the perception of Omomyc as a valid therapeutic agent has been often questioned. In this study, we used a mass spectrometry approach to evaluate the stability of Omomyc in tumor biopsies from murine xenografts following its intravenous administration. Our data strongly support that the integrity of the functional domains of Omomyc (DNA binding and dimerization region) remains preserved in the tumor tissue for at least 72 hours following administration and that the protein shows superior pharmacokinetics in the tumor compartment compared with blood serum. 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