In the rapidly evolving landscape of RNA therapeutics, achieving precise organ-specific delivery remains the most significant hurdle. At CD BioGlyco, we specialize in overcoming this challenge through our advanced GalNAc-miRNA delivery service. By leveraging the high affinity between N-acetylgalactosamine (GalNAc) and the asialoglycoprotein receptor (ASGPR) expressed abundantly on hepatocytes, we provide a robust platform for the targeted delivery of microRNA (miRNA) mimics and antagomirs directly to the liver.
This receptor-mediated endocytosis pathway offers an unparalleled advantage for treating hepatic diseases. While traditional delivery methods like lipid nanoparticles (LNPs) often face challenges with systemic toxicity or off-target accumulation, GalNAc conjugation provides a "lock-and-key" precision. This service is a critical component of our broader Therapeutic Nucleic Acid Development Platform, designed to assist pharmaceutical and biotech researchers in translating miRNA-based discoveries into viable clinical candidates.
We utilize a trivalent GalNAc architecture that exponentially increases the binding affinity to the ASGPR compared to monovalent ligands. This multivalent interaction ensures rapid and efficient internalization of the miRNA cargo into the hepatocyte cytoplasm.
The choice of linker is vital for the release and activity of the miRNA. We offer a variety of biolabile and stable linkers, optimized to withstand systemic circulation while ensuring the timely release of the oligonucleotide once inside the target cell.
To prevent premature degradation by nucleases, we employ a suite of chemical modifications including 2'-O-methyl (2'-OMe), 2'-fluoro (2'-F), and phosphorothioate (PS) linkages. These modifications enhance the pharmacokinetic profile without compromising the biological activity of the miRNA.
CD BioGlyco provides a comprehensive, end-to-end solution for liver-targeted miRNA delivery. As part of our therapeutic oligonucleotide delivery service, we address the unique requirements of each project, from initial design to large-scale synthesis.
We begin with an in-depth technical discussion to understand your therapeutic goals. Our experts evaluate the miRNA sequence, target gene, and required dosage to recommend the most effective GalNAc delivery strategy.
Utilizing advanced bioinformatics, we design the miRNA duplex or antagomir sequence. We integrate specific chemical modifications (e.g., 2'-OMe or 2'-F) tailored to enhance stability and minimize immunogenicity while maintaining silencing potency.
Both the sense and antisense strands are synthesized using automated solid-phase synthesis. We maintain stringent environmental controls to prevent contamination and ensure the highest possible crude yield.
The trivalent GalNAc ligand is conjugated to the optimized oligonucleotide using specialized click chemistry or phosphoramidite methods. This step is carefully monitored to ensure quantitative conversion and structural integrity.
The crude GalNAc-miRNA conjugate is purified via preparative HPLC or ion-exchange chromatography. We then perform multi-dimensional QC, including MALDI-TOF MS and endotoxin testing, to confirm identity and safety.
Upon request, we perform in vitro validation to confirm ASGPR-mediated uptake and gene knockdown. Finally, a detailed certificate of analysis (CoA) and all experimental data are provided along with the lyophilized product.
DoI: 10.3390/pharmaceutics13060817
Journal: Pharmaceutics
IF: 5.5
Published: 2021
Results: This article explores enhancing the in vivo activity of anti-miRNA-122 tiny locked nucleic acid (LNA) antisense oligonucleotides (ASO) via GalNAc conjugation. Tiny LNAs, though active and specific, have poor pharmacokinetics. The study develops GalNAc-conjugated constructs, with tL-5G3 showing an ED50 of 12 nmol/kg, 300-500 times more potent than the unconjugated version. GalNAc conjugation improves liver targeting, reducing kidney accumulation and toxicity. Optimal results are achieved by attaching GalNAc to the 5′ terminus via a biolabile phosphodiester bond. Dose-response and biodistribution studies confirm enhanced potency and favorable tissue localization. This strategy addresses tiny LNA's limitations, paving the way for clinical application of miRNA-targeting small ASO therapy.
Fig.1 In vivo dose-responsiveness study of tL-5G3. (Yamamoto, et al., 2021)
Metabolic Disorders
By delivering miRNA mimics or antagomirs to hepatocytes, researchers can address non-alcoholic fatty liver disease (NAFLD) and hyperlipidemia with high precision.
Hepatocellular Carcinoma
This approach modulates oncogenic pathways while sparing non-hepatic tissues, minimizing systemic side effects, and improving the therapeutic index of experimental RNA-based oncology treatments.
Viral Hepatitis Research
We help clients design conjugates that inhibit viral replication or enhance the host immune response specifically within the liver, offering a potent strategy for viral hepatitis research.
Genetic Liver Diseases
CD BioGlyco supports the development of therapies for urea cycle disorders and porphyria, providing stable and efficient tools for long-term gene regulation studies.
Exceptional Targeting Specificity
Our trivalent GalNAc clusters exhibit nanomolar affinity for the ASGPR exclusively on hepatocytes. This ensures that miRNA cargo is delivered with exceptional precision to the liver.
Enhanced Metabolic Stability
We employ advanced chemical modifications, including 2'-OMe and phosphorothioate linkages, to protect GalNAc-miRNA conjugates from nuclease degradation.
Reduced Systemic Toxicity
By utilizing receptor-mediated endocytosis, our delivery system avoids the immunogenicity and cytotoxicity often associated with viral vectors or lipid nanoparticles.
Proven Glycobiology Expertise
Our multidisciplinary team provides end-to-end support, from initial design to final validation, ensuring that every GalNAc-miRNA conjugate meets the highest scientific standards.
"The GalNAc-conjugated oligos showed remarkable stability and liver specificity in our mouse models, far exceeding the performance of our previous LNP formulations. The technical support team was incredibly knowledgeable about ASGPR kinetics."
– J.T., Senior Scientist
"The purity of the GalNAc-miRNA conjugates provided by CD BioGlyco is exceptional. We received a detailed CoA with LC-MS data that gave us full confidence in the material. Their ability to handle complex chemical modifications helped us achieve a sustained gene knockdown that lasted for weeks."
– A.D., Principal Investigator
"CD BioGlyco's one-stop solution for synthesis and conjugation saved our team months of optimization. Their trivalent GalNAc cluster technology provided the high affinity we needed to reduce our effective dose significantly."
– S.D., Director of R&D
CD BioGlyco is dedicated to advancing the field of RNA therapeutics by providing precision-engineered GalNAc-miRNA delivery services. By combining our deep roots in glycobiology with state-of-the-art oligonucleotide chemistry, we empower researchers to target liver-specific pathways with unprecedented accuracy and safety. Please feel free to contact us to discuss your sequence design, scale requirements, and delivery strategies.
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