Abstract
Targeted therapies are rapidly evolving modalities of cancer treatment. The largest group of currently developed biopharmaceuticals is antibody-drug conjugates (ADCs). Here, we developed a new modular strategy for the generation of cytotoxic bioconjugates, containing a homodimer of targeting protein and two highly potent anticancer drugs with distinct mechanisms of action. Instead of antibody, we applied human fibroblast growth factor 2 (FGF2) as a targeting protein. We produced a conjugate of FGF2 with either monomethyl auristatin E (MMAE) or α-amanitin (αAMTN) as a cytotoxic agent and subsequently applied a sortase A-mediated ligation to obtain a dimeric conjugate containing both MMAE and αAMTN. The developed method ensures site-specific conjugation and a controlled drug-to-protein ratio. We validated our approach by demonstrating that dimeric dual warhead conjugate exhibits higher cytotoxic potency against fibroblast growth factor receptor-positive cell lines than single-warhead conjugates. Our modular technology can be applied to other targeting proteins or drugs and thus can be used for preparation of different bioconjugates.
Keywords:
FGF2; MMAE; cancer; conjugates; sortase A; α-amanitin.
Publication types
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Research Support, Non-U.S. Gov't
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Validation Study
MeSH terms
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Alpha-Amanitin / chemistry*
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Aminoacyltransferases / chemistry
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Antineoplastic Agents / chemistry*
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Antineoplastic Agents / pharmacology
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Antineoplastic Agents / therapeutic use
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Bacterial Proteins / chemistry
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Cell Line, Tumor
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Cell Survival / drug effects
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Chemistry, Pharmaceutical / methods*
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Cysteine Endopeptidases / chemistry
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Dimerization
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Drug Screening Assays, Antitumor
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Fibroblast Growth Factor 2 / chemistry*
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Fibroblast Growth Factor 2 / isolation & purification
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Fibroblast Growth Factor 2 / pharmacology
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Fibroblast Growth Factor 2 / therapeutic use
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Humans
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Neoplasms / drug therapy*
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Neoplasms / pathology
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Oligopeptides / chemistry*
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Oligopeptides / pharmacology
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Oligopeptides / therapeutic use
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Receptor, Fibroblast Growth Factor, Type 1 / antagonists & inhibitors
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Receptor, Fibroblast Growth Factor, Type 1 / metabolism
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / pharmacology
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Recombinant Proteins / therapeutic use
Substances
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Alpha-Amanitin
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Antineoplastic Agents
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Bacterial Proteins
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Oligopeptides
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Recombinant Proteins
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Fibroblast Growth Factor 2
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Aminoacyltransferases
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sortase A
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FGFR1 protein, human
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Receptor, Fibroblast Growth Factor, Type 1
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Cysteine Endopeptidases
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monomethyl auristatin E