Targeted EpCAM-binding for the development of potent and effective anticancer proteins.

Area of Science:

• Biotechnology
• Oncology
• Protein Engineering

Background:

• Protein-based cancer therapies offer low toxicity but face absorption and stability challenges.
• Existing protein therapies require high doses and have slow-acting bioactivity.

Purpose of the Study:

• To develop a non-invasive, highly potent, and tumor-specific protein-based anticancer therapy.
• To overcome the limitations of conventional protein therapeutics for cancer treatment.

Main Methods:

• Conjugation of designed ankyrin repeat proteins (DARPin) with an anticancer protein to target epithelial cell adhesion molecule (EpCAM).
• In vitro assessment of anticancer efficacy and IC50 values of the DARPin-anticancer protein conjugate (drtHLF4).
• In vivo evaluation of oral and intratumoral administration of drtHLF4 in a HT-29 cancer murine model.

Main Results:

• DARPin-anticancer proteins demonstrated over 100-fold in vitro anticancer efficacy against EpCAM-positive cells within 24 hours.
• The drtHLF4 conjugate exhibited nanomolar IC50 values, indicating high potency.
• Orally administered drtHLF4 was absorbed systemically and cleared colorectal tumors in mice with a single dose.
• Intratumoral injection of drtHLF4 cleared subcutaneous tumors within three doses.

Conclusions:

• Developed a non-invasive, orally available protein-based cancer therapy with improved potency and tumor specificity.
• The DARPin-anticancer protein conjugate approach effectively addresses limitations of traditional protein-based therapies.
• This strategy offers a promising alternative for cancer treatment with enhanced efficacy and reduced dosage requirements.