Oral delivery of deoxycholic acid -coated lipid nanoparticles with anti-inflammatory drugs for the treatment of acute ulcerative colitis = 궤양성 대장염 치료를 위한 항염증제 함유 담즙산 코팅 지질나노입자의 경구 약물 전달|RISS 상세보기

다국어 초록 (Multilingual Abstract)

Ulcerative colitis (UC) is an unidentified colorectal inflammatory response syndrome. Colitis treatment involves administration combined in a patient-specific manner to determine the appropriate type of reduction in the inflammatory response. In this regard, it is necessary to quickly lower the level of pro-inflammatory cytokines that can cause inflammation in colon tissue. In the inflammatory mechanism, tumor necrosis factor-alpha (TNF-α), produced by macrophages, plays an important role in infiltrating the infected site and triggers the secretion of inflammatory cytokines. However, commercially available TNF-α antibody drugs have limitations such as fibrosis, cytokine storms, and high production costs. The need for UC treatments without side effects related to systemic control is increasing. For this reason, DOCA-coated lipid nanoparticles can be an innovative strategy for transferring drugs to colon tissues. It is a good platform for transferring desired drugs by controlling the composition of lipid nanoparticles. It was intended to deliver siRNA by oral administration by consisting of stable lipid nanoparticles at several pHs. In particular, drugs, which are TNF-α conversion enzymes (TACE), can be an innovative target for breaking the positive feedback loop of inflammatory mediators (TNF-α) because they convert inactive TNF-α membrane binding forms into active soluble forms in macrophages. In this study, a non-viral gene delivery system was developed to deliver siRNA to inflammatory-mediated macrophages without toxicity. Peptide-based gene carriers are produced by nine positively charged arginine (PAs-s). Our results showed that the siTACE/PAs-s complex was internalized in intestinal epithelial cells and filtered macrophages and successfully downregulated TACE mRNA levels. Finally, RNA interference in cell-targeted peptide carriers represents a fundamental treatment for acute inflammatory colitis with no off-target effect.

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목차 (Table of Contents)

1. Introduction 1
2. Material and Methods 5
2.1 Materials 5
2.2 Preparation of siRNA/Glycyrrhizin loaded deoxycholic acid-coated lipid nanoparticles 6
2.3 Zeta-average and zeta-potential measurement 6

1. Introduction 1
2. Material and Methods 5
2.1 Materials 5
2.2 Preparation of siRNA/Glycyrrhizin loaded deoxycholic acid-coated lipid nanoparticles 6
2.3 Zeta-average and zeta-potential measurement 6
2.4 pH stability Test 7
2.5 Cryo-TEM 7
2.6 In vitro co-culture model 8
2.7 Intracellular uptake mechanism test using flow cytometry 8
2.8 In vitro mRNA and protein expression level measurement 9
2.9 DSS-induced colitis model and oral administration of drugs 9
2.10 Biodistribution 10
2.11 Ex vivo analysis of mRNA expression 10
2.12 Ex vivo analysis of cytokines 11
2.13 Immunofluorescence analysis of colon 11
2.14 NADPH oxidase activity 12
2.15 MPO assay 12
2.16 Statistical analysis 13
3. Results 14
3.1 Synthesis and Characterization of particles 14
3.2 In vitro cellular uptake of GDNP(siTACE/PAs-s) in caco-2/RAW 264.7 indirect co-culture model 18
3.3 Anti-inflammatory effects of GDNP(siTACE/PAs-s) in caco-2 and raw264.7 cells 23
3.4 In vivo biodistribution of GDNP(siTACE/PAs-s) in the mouse GI tract 26
3.5 In vivo therapeutic efficacy of GDNP(siTACE/PAs-s) in mice with DSS-induced colitis 29
3.6 In vivo Immuno-regulatory effects of GDNP (siTACE/PAs-s) in mice with DSS-induced colitis 32
4. Conclusion 35
5. References 36
국문 요지 41