Development of Non-viral delivery system for gene therapy

Abstract

표적세포에 유전물질을 전달하기 위한 효율적이고 안전한 비-바이러스성 전달체의 개발은 임상 적용에서 매우 중요한 과제이다. 비-바이러스 전달체로써 2개의 유전자 전달체, CBA-106 polymer 와 102-C10 lipidoid가 개발되었다. 플라즈미드 DNA를 효율적으로 전달하기 위해 이황화 구조를 갖는 CBA와 34개의 아민을 사용해서 생분해성 poly(amido amine)s을 만들었다. 스크리닝 한 후에, 선도물질을 선택하고 특성을 분석하였다. CBA-106 polymer는 생리적 조건 하에서 높은 혈청 안정성 및 GSH에 의한 생체 내 분해 능을 가지며 높은 세포 생존력을 갖는다. 체내로 mRNA를 전달하기 위하여 8개의 amine과 4개의 epoxide를 사용해서 lipid 와 유사한 물질을 만들었고, 이 것을 lipidoid라고 부른다. 스크리닝 후에, 선도물질로 102-C10을 선택했고 다른 리포좀 제형성분인 DSPC, PEG 및 cholesterol과 혼합되어 mRNA가 표적 조직으로 잘 전달 될 수 있고 생체 내에서 단백질이 발현될 수 있음을 보였다. 이러한 결과로부터, CBA-106 과 102-C10은 유전자 치료를 위한 새로운 전달체로서의 잠재력이 있음을 결론 지을 수 있다.;To deliver genetic materials into target cells, the development of efficient and safe non-viral carrier is highly important for clinical application. As novel non-viral delivery systems, we report two different gene delivery carriers : CBA-106 bioreducible polymer and 102-C10 lipidoid. To efficiently deliver plasmid DNA, we prepared bioreducible poly(amido amine)s by reacting cystamine bisacrylamide (CBA) with 34 different small amines. After screening, we selected lead substances and characterized. CBA-106 polymer had high serum stability and bioreducible ability under physiological conditions and showed relatively low cytotoxicity. To deliver mRNA for in vivo applications, we synthesized lipid-like materials, called the lipidoids using 8 of small amines and 4 of epoxide lipids. After screening, we selected lead lipidoids molecules, 102-C10. Our lead compound, 102-C10, was mixed with other liposome formulating components to efficiently deliver mRNA in vivo. Formulation having DSPC, PEG-ceramide, and lipidoids showed that mRNA can be well delivered to the target tissue and protein expression can be achieved in vivo. From our results, we can conclude that the lead substances such as CBA-106 and 102-C10 have great potential as a novel delivery carrier for gene therapy applicationsTo deliver genetic materials into target cells, the development of efficient and safe non-viral carrier is highly important for clinical application. As novel non-viral delivery systems, we report two different gene delivery carriers : CBA-106 bioreducible polymer and 102-C10 lipidoid. To efficiently deliver plasmid DNA, we prepared bioreducible poly(amido amine)s by reacting cystamine bisacrylamide (CBA) with 34 different small amines. After screening, we selected lead substances and characterized. CBA-106 polymer had high serum stability and bioreducible ability under physiological conditions and showed relatively low cytotoxicity. To deliver mRNA for in vivo applications, we synthesized lipid-like materials, called the lipidoids using 8 of small amines and 4 of epoxide lipids. After screening, we selected lead lipidoids molecules, 102-C10. Our lead compound, 102-C10, was mixed with other liposome formulating components to efficiently deliver mRNA in vivo. Formulation having DSPC, PEG-ceramide, and lipidoids showed that mRNA can be well delivered to the target tissue and protein expression can be achieved in vivo. From our results, we can conclude that the lead substances such as CBA-106 and 102-C10 have great potential as a novel delivery carrier for gene therapy applications.