Synthesis and evaluation of antisense oligonucleotides prodrug with G-quadruplex assembly and lysosome escape...

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Delivery

Background:

• Antisense oligonucleotides (ASOs) are potent therapeutic agents for various diseases.
• Challenges in ASO therapy include poor stability and bioavailability, limiting clinical efficacy.
• Existing delivery methods often struggle with cellular uptake and endosomal entrapment.

Purpose of the Study:

• To develop a novel antisense oligonucleotide (ASO) prodrug with enhanced stability and cellular delivery.
• To investigate the potential of G-quadruplex assembly and lysosome escape mechanisms for improved oncotherapy.
• To evaluate the therapeutic efficacy of the optimized G-quadruplex-ASO conjugate in preclinical cancer models.

Main Methods:

• Design and synthesis of a G-quadruplex-forming ASO prodrug conjugated with maleimide.
• Assessment of enzymatic degradation resistance and cellular uptake efficiency in vitro.
• Evaluation of therapeutic effects in vitro and in vivo using cancer models.

Main Results:

• The end-assembled G-quadruplex structure significantly protected the ASO from enzymatic degradation.
• Maleimide conjugation facilitated enhanced cellular uptake, potentially bypassing lysosomal pathways.
• The optimized Mal²-G4-ASO molecule demonstrated significant therapeutic efficacy in both in vitro and in vivo cancer models.

Conclusions:

• This G-quadruplex-ASO prodrug strategy offers a promising approach to overcome ASO stability and delivery challenges.
• The developed prodrug exhibits enhanced anticancer activity through improved cellular internalization and resistance to degradation.
• This work paves the way for advancing nucleic acid-based therapies in oncology and other disease areas.