Design, Synthesis, Molecular Docking, and Preclinical Evaluation of a New Radiolabeled PEG3-Linked FAPI Derivative for Fibroblast Activation Protein Targeting

AuthorMahshid Kianien
AuthorMehdi Akhlaghien
AuthorSafura Jokaren
AuthorOmid Bavien
AuthorHooman Hafezien
AuthorKhosrou Abdien
AuthorOmid Sabzevarien
AuthorSaeed Balalaieen
AuthorFarhad Golmohammadien
AuthorZahra Ghiamatyen
AuthorSara Roustaeien
AuthorAlireza Foroumadien
AuthorDavood Beikien
OrcidSaeed Balalaie [0000-0002-5764-0442]en
OrcidDavood Beiki [0000-0002-2862-0581]en
Issued Date2026-12-31en
AbstractBackground: Fibroblast activation protein (FAP) is a promising molecular target for cancer theranostic applications. However, current fibroblast activation protein inhibitors (FAPIs) have limitations, including rapid clearance and limited tumor retention. Objectives: This study aimed to develop a novel PEG3-linked FAPI derivative, [68Ga]Ga-FAPI-MKG, with enhanced tumor accumulation and retention. FAPI-MKG was prepared by incorporating a PEG3 linker into the FAPI-04 structure to optimize its pharmacokinetic profile. Methods: The compound was synthesized via an 11-step route starting from quinine sulfate and radiolabeled with gallium-68. In vitro studies included determination of lipophilicity (Log P) and stability assays in saline and human serum albumin. Preclinical evaluation in BALB/c mice bearing CT-26 tumors included biodistribution, blocking studies, and PET/CT imaging. Molecular docking and molecular dynamics simulations were performed to provide mechanistic insights into binding interactions. Results: [68Ga]Ga-FAPI-MKG was successfully synthesized with high radiochemical purity (> 98%) and a molar activity of 414.79 mCi/μmol. It demonstrated moderate hydrophilicity (Log P = -3.26 ± 0.18) compared with the reference radiotracer, [68Ga]Ga-FAPI-46 (-3.58 ± 0.29), and high stability (RCP > 90% after 120 minutes). In vivo studies showed significantly higher tumor uptake (7.18 ± 0.56% ID/g at 60 minutes; 3.20 ± 0.11% ID/g at 120 minutes) and prolonged retention compared with the reference radiotracer, along with dual hepatobiliary and renal excretion pathways. Blocking studies confirmed FAP-specific uptake. Computational analyses indicated strong binding energy (-9.8 kcal/mol) and optimized electrostatic interactions with FAP. The strategic incorporation of a PEG3 linker into [68Ga]Ga-FAPI-MKG significantly improved tumor accumulation, extended tumor retention, and increased the tumor-to-background ratio. Conclusions: These findings suggest that [68Ga]Ga-FAPI-MKG may be a promising candidate for clinical translation for imaging and theranostics of FAP-expressing cancers.en
DOIhttps://doi.org/10.5812/ijpr-170734en
URIhttps://brieflands.com/journals/ijpr/articles/170734en
KeywordCancer-associated Fibroblastsen
KeywordGallium-68en
KeywordMolecular Imagingen
KeywordMolecular Dynamicsen
KeywordPET Imagingen
KeywordRadiolabelingen
PublisherBrieflandsen
TitleDesign, Synthesis, Molecular Docking, and Preclinical Evaluation of a New Radiolabeled PEG3-Linked FAPI Derivative for Fibroblast Activation Protein Targetingen
TypeResearch Articleen

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