NATURAL APPROACH

Our own laboratory allows the development
of nuclear microRNA drugs for RNA activation of therapeutic genes from discovery to
pre-clinical proof-of-concept.

RNatives developed a novel discovery pipeline for the identification of the drug candidates. Our approach is science-driven and based on years of research on RNA activation. During multiple steps of different types of analysis in silico, we narrow down the best targets and their prime regulators. These steps include:

• Identification of the key target genes for each indication

• In silico analysis of the feasibility of the selected target genes by analysis of target genes regulatory region

• Rational discovery/design of the target gene modulators by an in-house developed proprietary software

• Selection of best RNA drug leads for testing in the wet lab

The best RNA drug candidates are synthesized and taken into in vitro testing in our own BSL2-level cell culture and molecular biology laboratories. At this point, we test the functionality of the lead microRNAs in relevant cell culture models, followed by modern molecular biology methods, including:

• MicroRNA drug leads are tested in relevant cell culture models by knock-in/out analyses

• Target protein expression is verified

• Next-generation sequencing analysis is done to verify the specificity of RNA activation

• Confirmed leads are taken forward to formulation selection

RNatives has long experience in working with extracellular vesicles (EVs), such as exosomes. These vesicles are natural carriers of molecular messages between the tissues in the body, and as such are a natural choice for introducing candidate drugs microRNAs into cells. RNatives developed a process for R&D level production and downstream purification of EVs. The key steps are:

• The stable cell line is produced to overexpress microRNA for loading to EVs

• Upstream production of EVs, small or large scale

• Downstream purification and concentration of EVs

• Analysis and quality control of EV/miRNA product

• Functional testing of EV/miRNA drug in target cells (release and potency)

Combining in silico and in vitro work results in a confined number of leads, that are scientifically relevant to the desired indication and have shown functional effects in relevant cell lines. RNatives work together with trusted, validated, and high-quality contract research organizations (CROs) to shift the initial molecular findings into physiological, meaningful results in a disease model. These proofs-of-concept (POC) experiments drive the selection of RNatives drugs towards IND-enabling studies.

After POC studies in animals such as mouse or rabbit, RNatives is fully prepared to take the next steps from the lab to the clinic. We work with experts and consultants from regulatory and clinical fields to make the transformation of the lead miRNA molecule into actual therapy. These processes include:

• Safety, toxicology, and dose escalation studies are done in relevant species

• IND-application is submitted to a regulatory authorities