Application of genetic constructions as novel generation of medicines in nearest future will enable the effective treatment of different genetic disorders as well as curing of serious social diseases, such as cancer and diabetes. The prospective therapeutic medicines can be developed as based on small interfering RNAs (siRNAs), plasmid DNAs (pDNAs) and matrix RNAs (mRNAs). First ones could be used to effectively “switch of” the expression of undesired protein, while later ones are useful for induction (“switch on”) of protein expression.
The instability of nucleic acids (NAs) in the physiological environment as well as their toxic effects on healthy cells hinder the application of genetic drugs. Thus, the development of the delivery systems, which protect NAs and enhance the efficiency of their intracellular internalization into the target cells, is obviously necessary. It should be also noted, that the milestone of the gene delivery systems is the possibility of their systemic in vivo administration, which should make them commercially applicable.
In this work, we develop nanogels for NAs delivery based on interpolyelectrolyte complexes (IPECS), which are cross-linked with a pH-responsive ketal linker.
This study continues our previous G-RISC research project of Marina Khazanova, which was titled «pH-sensitive interpolyelectrolyte poly-L-lysine/heparin complexes for intracellular delivery of nucleic acids».
The following results have been achieved during our previous work:
1.The pH-sensitive ketal linker N,N'-(propane-2,2-diyl bis (oxy))-bis-(-ethane-2,1-diyl))bis(2-methacrylamide) was obtained. The degradation of the linker at acidic рН was confirmed (LabBiomedChem IC SPbU).
2.The procedure of obtaining the cross-linked pLys/Hep nanogels through photoinduced thiol-ene reaction was developed (LabBiomedChem IC SPbU).
3.The size and morphology of the nanogel particles were determined by TEM and DLS methods (LabBiomedChem IC SPbU).
4.The kinetic of pH-dependent release was evaluated. The increased release of the model oligonucleotide in acidic media was demonstrated (LabBiomedChem IC SPbU).
5.The lack of cytotoxicity of cross-linked nanogel particles was demonstrated on BEAS 2B, A549, HEK 293 и NIH 3T3 cell lines (ITC UH).
6.The possibility of using the cross-linked nanogel particles for transfection of small interfering RNA, which knocks down expression of green fluorescent protein, was demonstrated (ITC UH).