Lipid nanoparticles(LNPs)are critical for the delivery of drugs and nucleic acids.However,current mRNA-LNP formulations require stringent freezing for storage,which limits their global distribution.Our previous studie...Lipid nanoparticles(LNPs)are critical for the delivery of drugs and nucleic acids.However,current mRNA-LNP formulations require stringent freezing for storage,which limits their global distribution.Our previous studies demonstrated that optimizing the lipid type or molar ratio of Comirnaty-type mRNA-LNPs could enhance their lyophilization stability,thus improving their long-term storage stability under mild conditions.This study aims to enhance the storage stability of Spikevax-type mRNA-LNPs by optimizing lipid compositions and utilizing lyophilization for storage at 4�C.Fifteen mRNA-LNP formulations were evaluated for their physicochemical properties and transfection efficiency(TE)in human embryonic kidney(HEK)-293T cells using the I-optimal design of mixture experiments.Mathematical models were developed to predict the relationships among encapsulation efficiency,transfection performance and lipid ratios.The optimized mRNA-LNP formulation(N4),with a 1,2-distearoyl-sn-glycero-3-phosphocholine(DSPC)-to-cholesterol ratio of 0.36,exhibited superior stability and TE after lyophilization.N4 outperformed the original Spikevax formulation in several cell lines,including eye-derived ARPE-19 cells and lung-derived A549 cells.In vivo,N4 demonstrated high TE in the spleen of C57BL/6 mice both before and after lyophilization,with no signals observed in the kidneys,heart or eyes.These findings suggest that the optimized N4 formulation offers a robust,stable and efficient delivery system for gene therapy and vaccines,potentially overcoming the storage limitations of current Spikevax-type mRNA-LNPs and broadening their therapeutic applications.展开更多
基金supported by the Zhejiang Provincial Natural Science Foundation of China(LQ24H120006)the Wenzhou Municipal Science and Technology Bureau(Y2023799)+3 种基金the Startup Foundation for Scientific Research,Eye Hospital,Wenzhou Medical University(KYQD20211204)the National Natural Science Foundation of China(52250710155)the National Key Research and Development Program of China(2021YFA1101200,2022YFA1105501)the Project of State Key Laboratory of Ophthalmology,Optometry and Visual Science,the Wenzhou Medical University(J02-20210201).
文摘Lipid nanoparticles(LNPs)are critical for the delivery of drugs and nucleic acids.However,current mRNA-LNP formulations require stringent freezing for storage,which limits their global distribution.Our previous studies demonstrated that optimizing the lipid type or molar ratio of Comirnaty-type mRNA-LNPs could enhance their lyophilization stability,thus improving their long-term storage stability under mild conditions.This study aims to enhance the storage stability of Spikevax-type mRNA-LNPs by optimizing lipid compositions and utilizing lyophilization for storage at 4�C.Fifteen mRNA-LNP formulations were evaluated for their physicochemical properties and transfection efficiency(TE)in human embryonic kidney(HEK)-293T cells using the I-optimal design of mixture experiments.Mathematical models were developed to predict the relationships among encapsulation efficiency,transfection performance and lipid ratios.The optimized mRNA-LNP formulation(N4),with a 1,2-distearoyl-sn-glycero-3-phosphocholine(DSPC)-to-cholesterol ratio of 0.36,exhibited superior stability and TE after lyophilization.N4 outperformed the original Spikevax formulation in several cell lines,including eye-derived ARPE-19 cells and lung-derived A549 cells.In vivo,N4 demonstrated high TE in the spleen of C57BL/6 mice both before and after lyophilization,with no signals observed in the kidneys,heart or eyes.These findings suggest that the optimized N4 formulation offers a robust,stable and efficient delivery system for gene therapy and vaccines,potentially overcoming the storage limitations of current Spikevax-type mRNA-LNPs and broadening their therapeutic applications.
