mRNA Pharmacology Optimization

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.

Fig.1 Structural features of IVT mRNA. (Verbeke, 2019)

mRNA Pharmacology Optimization

• Optimization Service to Improve mRNA Translation and Stability
1. 5'-cap modifications:
• Uncapped, functional when combined with IRES (internal ribosome entry site)
• Cap analogues mediating binding to eIF4E (eukaryotic translation initiation factor 4E)
• Cap analogues conferring resistance to decapping
2. Coding region:
• Optimized codon usage to improve the translation
• Optimized base usage to reduce the endonucleolytic attack
3. Poly(A) tail:
• Masked/unmasked poly(A) tail affecting translation
• Length of poly(A) tail affecting stability
• Modified nucleotides inhibiting deadenylation
4. 5' UTR:
• Regulatory sequence elements binding to molecules involved in mRNA trafficking and translation
• Sequences inhibiting-exonucleolytic degradation
5. 3' UTR:
• Sequence elements mediating binding to proteins involved in mRNA trafficking and translation
• Sequences repressing deadenylation of mRNA
6. Whole IVT mRNA:
• Use of modified nucleosides for modulating innate immune activation
• 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
  Fig.2 Nanoscale platforms for mRNA delivery. (Li, 2019)
• 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.

Reference

1. Verbeke, R.; et al. Three decades of messenger RNA vaccine development. Nano Today. 2019, 28: 100766.
2. Li, B.; et al. Nanoscale platforms for messenger RNA delivery. Wiley Interdisciplinary Reviews: Nanomedicine and Nanobiotechnology. 2019, 11(2): e1530.