mRNA Pharmacology Optimization

Background

In vitro transcribed (IVT) messenger RNA (mRNA) has recently come into focus as a potential new drug class to deliver genetic information. Such synthetic mRNA can be engineered to transiently express proteins by structurally resembling natural mRNA. Armed with advanced mRNA technology, Creative Biolabs is devoted to offering mRNA pharmacology optimization services as either individual service or part of a complete project for global customers.

• Introduction of mRNA Therapy

The concept behind using IVT mRNA as a drug is the transfer of a defined genetic message into the cells for the ultimate purpose of preventing or altering a particular disease state. One method is to transfer mRNA into the patient's cells ex vivo. These transfected cells are then adoptively administered back to the patient. The other is the direct delivery of IVT mRNA using various routes. Fig.1 shows the structural elements of IVT mRNA. Each one can be optimized and modified to modulate the stability, translation capacity, and immune-stimulatory profile of mRNA.

mRNA Pharmacology Optimization

• Optimization Service to Improve mRNA Translation and Stability
  • 5'-cap modifications:
    1. Uncapped, functional when combined with IRES (internal ribosome entry site)
    2. Cap analogues mediating binding to eIF4E (eukaryotic translation initiation factor 4E)
    3. Cap analogues conferring resistance to decapping
  • Coding region:
    1. Optimized codon usage to improve the translation
    2. Optimized base usage to reduce the endonucleolytic attack
  • Poly(A) tail:
    1. Masked/unmasked poly(A) tail affecting translation
    2. Length of poly(A) tail affecting stability
    3. Modified nucleotides inhibiting deadenylation
  • 5' UTR:
    1. Regulatory sequence elements binding to molecules involved in mRNA trafficking and translation
    2. Sequences inhibiting-exonucleolytic degradation
  • 3' UTR:
    1. Sequence elements mediating binding to proteins involved in mRNA trafficking and translation
    2. Sequences repressing deadenylation of mRNA
  • Whole IVT mRNA:
    1. Use of modified nucleosides for modulating innate immune activation
    2. Engineering favorable secondary structures by sequence optimization
• Optimization Service for Immune-stimulatory Activity
  IVT mRNA can be created by incorporating naturally occurring modified nucleosides such as 2-thiouridine, pseudouridine, 5-methylcytidine, 5-methyluridine, or N6-methyladenosine into the IVT mRNA. This has been shown to suppress both the intrinsic adjuvant activity of IVT mRNA as well as its inhibitory effects on translation.
• Optimization Service for Immunogenicity Modulation
  mRNA is potentially beneficial for vaccination because it can provide adjuvant activity to drive dendritic cell (DC) maturation and thus elicits a strong sputum and B cell immune response. Our optimization services include the following aspects:
  • Nucleosides-modified mRNA
  • Sequence-optimized mRNA
  • Self-amplifying mRNA
  • HPLC (high-performance liquid chromatography)-purified
  • FPLC (fast protein liquid chromatography)-purified
  • Addition of adjuvant
• Optimization Service for Improved mRNA Delivery Efficiency
  There are two challenges associated with the delivery of IVT mRNA: one is to achieve a sufficiently high net level of the encoded protein and the other is to reach a high number of cells. In vivo delivery of mRNA into a high fraction of defined target cell populations is challenging and depends on the accessibility of target cells. Creative Biolabs found that multiple structural motifs of the amino lipid are important for efficient in vivo performance of mRNA-containing lipid-based nanoparticle (LNP), including surface charge, structure, and position of the ester in the lipid tails and structure of the head group. We have identified lipids that balance chemical stability, improved efficiency of delivery due to improved endosomal escape, rapid in vivo metabolism, and a clean toxicity profile.
  • Loading mRNA into DCs ex vivo followed by re-infusion of the transfected cells.
  • Direct parenteral injection of mRNA with or without a vector.
  • Various delivery systems especially in LNP formats.

For more detailed information, please feel free to contact us or directly send us an online inquiry.

Highlights

• Comprehensive mRNA Pharmacology Optimization: Creative Biolabs offers specialized mRNA pharmacology optimization services, focusing on enhancing mRNA translation, stability, and immune-stimulatory activity, ensuring high efficacy and safety in therapeutic applications.
• Advanced IVT mRNA Technology: Our cutting-edge IVT mRNA technology enables the engineering of synthetic mRNA to transiently express proteins, providing a novel approach to disease prevention and treatment by delivering genetic information directly into cells.
• Optimized mRNA Delivery Systems: Our expertise in lipid nanoparticle (LNP) formulations ensures efficient in vivo delivery of mRNA to target cells, overcoming challenges related to protein expression levels and cell targeting.
• High-Efficiency mRNA Delivery: Creative Biolabs has developed lipid-based nanoparticles with optimized surface charge and chemical stability, ensuring high delivery efficiency, improved endosomal escape, and a clean toxicity profile for mRNA therapies.
• Customizable mRNA Optimization Services: We provide tailored optimization services, including 5'-cap modifications, codon usage optimization, and poly(A) tail modifications, to improve the translation and stability of mRNA, ensuring maximum therapeutic effectiveness.

FAQs

Published Data

The study focused on optimizing mRNA pharmacology, particularly the non-coding regions of mRNA, to improve the efficacy of a non-modified mRNA COVID-19 vaccine. The research compared two versions of mRNA vaccines: CVnCoV and CV2CoV. The CV2CoV vaccine, which included optimized non-coding regions, demonstrated significantly better outcomes compared to CVnCoV. Specifically, CV2CoV induced higher levels of binding and neutralizing antibodies, stronger memory B cell and T cell responses, and provided more robust protection against SARS-CoV-2, including the Delta variant, in non-human primates. These results underscore the importance of optimizing mRNA's non-coding regions to enhance vaccine immunogenicity and protective efficacy, suggesting that such optimization can lead to more effective mRNA vaccines.

Fig.1 CV2CoV induces strong binding and neutralizing antibodies in macaques.¹

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Reference

1. Gebre, Makda S., et al. "Optimization of non-coding regions for a non-modified mRNA COVID-19 vaccine." Nature 601.7893 (2022): 410-414. Distributed under Open Access license CC BY 4.0, without modification.