Chemical coupling of an active/passive dual-targeting fusion protein ER(Fv)-HSA to DM1 improves its tumor-specificity and therapeutic efficacy against pancreatic cancer

Eur J Pharm Sci. 2025 Sep 1:212:107188. doi: 10.1016/j.ejps.2025.107188. Epub 2025 Jul 1.

Abstract

Owing to the lack of early diagnosis and radio/drug-resistance, the conventional therapies can not achieve ideal therapeutic effects on pancreatic cancer, thus targeted therapy with efficiency and specificity attracts more attention. In this study, a dual-targeting fusion protein ER(Fv)-HSA sequentially linked by anti-EGFR single-chain variable fragment, G4S linker and human serum albumin was constructed, and then was coupled with a derivative of maytansine DM1 using succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxylate linker for fusion protein-drug conjugate ER(Fv)-HSA-DM1, in which the affinity to tumor was preserved. With the help of active targeting of scFv and passive targeting of HSA, ER(Fv)-HSA-DM1 showed excellent tumor-targeting that rapidly accumulated at the tumor site and retained for a long time, moreover, part of conjugate was transported into the cytoplasm. At the same molar concentration, ER(Fv)-HSA-DM1 displayed stronger cytotoxicity than DM1 on pancreatic cancer cells, with the half-maximal inhibitory concentration value of 10-10-10-9 M, reflecting in microtubule networks destruction, cell cycle arrest and apoptosis. In AsPC-1 xenograft athymic mice models, the inhibition rate of DM1 at the dosage used equivalent to human clinical dosage was only 37.3 %, while the inhibition rate of ER(Fv)-HSA-DM1 loaded molar equivalent DM1 was 53.6 %, without any body weight loss. The results revealed that ER(Fv)-HSA effectively endowed DM1 with tumor-targeting and improved its antitumor efficacy, indicating active/passive dual-targeting fusion protein might be a feasible strategy for development of new targeted drugs for pancreatic cancer therapy.

Keywords: DM1; Epidermal growth factor receptor; Human serum albumin; Pancreatic cancer; Single-chain variable fragment.

MeSH terms

  • Animals
  • Antineoplastic Agents* / administration & dosage
  • Antineoplastic Agents* / chemistry
  • Antineoplastic Agents* / pharmacology
  • Antineoplastic Agents* / therapeutic use
  • Apoptosis / drug effects
  • Cell Line, Tumor
  • Female
  • Humans
  • Maytansine* / administration & dosage
  • Maytansine* / analogs & derivatives
  • Maytansine* / chemistry
  • Maytansine* / pharmacology
  • Maytansine* / therapeutic use
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Pancreatic Neoplasms* / drug therapy
  • Pancreatic Neoplasms* / metabolism
  • Pancreatic Neoplasms* / pathology
  • Recombinant Fusion Proteins* / administration & dosage
  • Recombinant Fusion Proteins* / chemistry
  • Serum Albumin, Human* / chemistry
  • Single-Chain Antibodies* / administration & dosage
  • Single-Chain Antibodies* / chemistry
  • Xenograft Model Antitumor Assays

Substances

  • Maytansine
  • Antineoplastic Agents
  • Serum Albumin, Human
  • Single-Chain Antibodies
  • Recombinant Fusion Proteins