Nucleic Acid Drug

Popular types

ASO
siRNA
mRNA
Aptamer
Advantages
Medicilon uses simple materials, convenient preparation processes and affordable production costs, toshorten the drug development cycle, making it possible to customize individual treatment plans. Hence, Medicilon offers aa excellent service solution for rare diseases.
Nucleic Acid Synthesis and Chemical Modifications
Synthesis
◆ Monomer synthesis
◆ Oligonucleotide synthesis
◆ Delivery system synthesis
◆ Oligo conjugate synthesis
Modification
◆ Sugar modification
◆ Nucleobase modification
◆ Backbone modification
◆ Delivery system modification
The Medicilon nucleotide drug R&D platform can provide a one-stop option for monomer synthesis and modification; oligonucleotide synthesis; delivery system synthesis and oligonucleotide conjugate synthesis. The siRNA library that has been built has a rich monomer inventory anda large monomer synthesis building block library, to support the synthesis of various modified monomers. Medicilon has a professional small nucleic acid drug R&D team that can provide efficient and fast R&D services; several siRNA and other oligonucleotide drug FTE projects have been completed and are in progress.
Bioactivity Screening
◆ Evaluation of binding between siRNA-GALNAc and targeted liver cells (ELISA, SPR, FP, FACS, MSD, Confocal microscope)
◆ Evaluation of decrease in target mRNA/protein level (RT-PCR, WB)
◆ Evaluation of cell phenotypes and functional regulation (Cell proliferation, Migration, Proteomics, and Transcriptome analysis).
◆ Evaluation of off-target activity
◆ Searching for potential off-target mRNA/protein in databases, such as NCBI, nucleotide BLAST.
◆ Unbiased analysis applying approaches including RNAseq, RNA Microarray, or targeted panel analysis using Nanostring.
Nucleic Acid Process R&D
Select starting materials: We aim to choose starting materials that are easy to purchase, possess mild toxicity and show good stability;Process R&D of nucleic acid:We aim to develop stable and green synthesis routes demonstrating low cost and high security.Quality Control:Up-to-date quality control system with complete technical standard.Scale up:End to end service ensuring smooth transfer.

Nucleic acid drugs preparation

Due to their low immunogenicity, biocompatibility and high encapsulation efficiency for oligonucleotide molecules, lipids and their derivatives have become a go-to delivery systems for nucleic acid drugs in recent years. The system is positively charged in the physiological environment. Negatively charged nucleic acid molecules are encapsulated by electrostatic action, and the positively charged surface can also help the entire carrier system to combine with the cell membrane of the target cell, thereby playing a delivery role.
Picture Source：Science China Life Sciences volume 62,pages333-348(2019)
Common delivery systems
Medicilon’s preparation methods of nanoparticles
Traits of successful delivery systems
Easily modified, easily synthesized, easily produced.The on-target and off-target ratio of delivery should be within an acceptable range.The effective dose must be significantly lower than the toxic dose.The bioactivity of the nucleic acid should be consistent from batch to batch.In most clinical cases, repeated administration does not result in loss of efficacy or safety.
Medicilon’s nanoparticle R&D lab
Nucleic acid-lipid system R&D
• Formulation: drug to lipid ratio, solvent screening, aqueous to organic solvent ratio
• Process: Preparation methods
• Stability
• Dosage form screening

Nucleic acid drugs bioanalysis

PK/TK analysis

• H-ELISA LC-MS/MS ；

• H-ECL；

• RT-qPCR;

• qPCR；

• ddPCR

ADA analysis

• Total ADA；

• MSD ；

• Nab analysis: CLBA or cell-based assay

PD or TOX related cytokines & biomarkers

Cytokine&Biomarker

• Singleplex；

• Multiplex (Luminex, MSD, FACS CBA)

• FACS

Medicilon case:Compound A –siRNA plasma quantification (20 µL plasma)

Solutions for Nucleic acid bioanalysis

LC-MS/MS/HRMS Platform
High specificityHigh sensitivity: ng levelAdvantages: end product detectable
Hybridization-EIA/ECL Platform
High specificitySensitivity: Detectable within 1 log copy Advantage：More Sensitive
qPCR/ddPCR Platform
Sensitivity: pM levelAdvantages: variable marking strategy; personalized reaction strategy.

Pharmacology evaluation

◆ Comparing different drug delivery methods

◆ Correlative analysis among nucleic acid drug pharmacology, target mRNA/protein degradation and drug PK

• Animals: Female BALB/c Nude mice；

• Cells: MDA-MB-231, 5x10⁶/mouse；

• Model Establishment: Right flank SC injection；

• Treatment: IV injection;

TIW (three times a week);

Group3, 4: mRNA (LNP)group.

• Animals: Female BALB/c Nude mice；

• Cells: MDA-MB-231, 5x10⁶/mouse；

• Model Establishment: Right flank SC injection；

• Treatment: Intratumor in-jection;

TIW (three times aweek);

Group 7, 8: mRNA(LNP) group.

◆ Dendrimer LNP

◆ Luc mRNA;Ⅳ and intra-tumoral

◆ CNP-generated exosome

◆ Delivery of PTEN mRNA by Glioma-directed Exosome EXO-T,IV

Syngeneic mouse models

Cancer Type	Cell Lines
Bladder Cancer	MB49
Brain Cancer	G261
Breast Cancer	4T1, EMT6, JC,EO771
Colon Cancer	CT26.WT, MC-38, Colon26
Leukemia	C1498, L1210, WEHI-3
Liver cancer	H22, Hepa 1-6
Lung Cancer	LLC1, KLN205
Lymphoma	A20, EL4, L5178-R, E.G7-OVA
Mastocytoma	P815
Melanoma	B16-F10, Clone-M3
Myeloma	J558
Pancreas Cancer	Panc 02
Renal Cancer	RENCA

Luciferase Cell Line

G261-luc, 4T1-luc, Mc38-luc, H22-luc, B16-F10-luc, LLC1-luc

Humanized mouse models

Cancer Type	Cell Lines in PBMC or HSC CD34+ Humanized Mice
Brain Cancer	U-87 MG
Breast Cancer	HCC1954, MDA-MB-231, JIMT-1
Colon Cancer	HT29, LoVo, Ls174T, HT-15
Gastric Cancer	NCI-N87, NUGC-4
Leukemia	THP-1
Lung Cancer	HCC827, NCI-H1975, NCI-H292, A549
Lymphoma	Raji, TMD8, MOLM-13
Melanoma	A375
Myeloma	RPMI-8226, NCI-H929, MM.1S
Ovarian Cancer	OVCAR-3
Pancreatic Cancer	Capan-2
Renal Cancer	786-O
Skin Cancer	A431

The Pharmacokinetics Research of Nucleic Acid Drugs

Medicilon Liver Biopsy Guided By B-ultrasound In Cynomolgus Monkeys Platform
The development of gene therapy and nucleic acid drugs has made the establishment of monkey models and related research a hot topic. Due to the similarity of genetic, morphological, physiological and biochemical characteristics with humans, non-human primates, especially cynomolgus monkeys, are closest to humans in terms of evolution, and have outstanding advantages for model construction, disease mechanism research, and drug development. Many disease models have been established so far.
The Medicilon Liver Biopsy Guided By B-ultrasound In Cynomolgus Monkeys Platform can avoid the large blood vessels and gallbladder and has the advantages of less trauma, safe and simple puncture operation, accurate positioning, and better postoperative recovery. Medicilon Liver Biopsy Guided By B-ultrasound In Cynomolgus Monkeys Platform can dynamically display the whole process of biopsy needle insertion and material collection in real time, which greatly improves the success rate of puncture and the accuracy of experimental results. Also, it can be used for the preclinical PK evaluation of gene therapy drugs, which promotes the improvement of experimental animal welfare, and provides accurate pathological basis for the dynamic monitoring and modeling progress of various liver disease models.

Learn more about "Nucleic Acid Drug R&D "

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FAQs

What are the Advantages and Disadvantages of Nucleic Acid Drugs?

Advantages:
Short development cycle and rapid target identification;
A wide range of treatment areas;
Resistant to drug resistance;
Long-lasting therapeutic effect;
High success rate in R&D
Disadvantages:
Lack of targeting by the drug itself;
Instability within the body, leading to easy clearance.
Preclinical Safety Characteristics of siRNA Drugs

• The biological effects of silencing of target mRNA may be present, but other effects are not easily observable.
• Due to the specific nature of siRNA sequence design, binding effects on non-target mRNAs are rare.
• Due to RISC’s high efficiency, siRNA rarely competes with other miRNA resources. Many toxic reactions associated with siRNA drug occur at the administration site, tissue, in tissue or cell residue or through accumulation in animals, including inflammatory reactions, hepatocyte vacuolation, renal tubular vacuolation, and deposition of basophilic granules etc.
• There are also some toxic reactions associated with the excretion process of siRNA drugs
• siRNA has the potential to induce innate and acquired immune responses, but this effect is often not evident in routine non-clinical safety studies.
• Immunogenicity may occur, but the antigen is not necessarily the siRNA itself; it can also be the excipients or structural components of the formulation. For example, if PEG is present, it can easily lead to the production of anti-PEG antibodies.
What are the Advantages and Disdvantages of RNA?

RNA: by degrading target mRNA
Advantage:
    Can be delivered to cross the cell membrane
    Can achieve high potencies for their targets
    siRNA molecules tend to exhibit longer persistence of target mRNA knockdown.
Disadvantage:
    High metabolic instability
    Off-target effects
    Poor cell permeability
    Low bioavailability, Hepatotoxicity, poor PK and limited tissue distribution, poor solubility and stability, primarily targets liver diseases

Nucleic Acid Drug

Nucleic Acid Drug

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