Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic Acids
Nucleic acids are promising for a variety of therapies, such as cancer therapy and the gene therapy of genetic disorders. The therapeutic efficacy of nucleic acids is reliant on the ability of their efficient delivery to the cytosol of the target cells. Amino lipids have been developed to aid in the...
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MDPI AG
2021-10-01
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| Series: | Pharmaceuticals |
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| Online Access: | https://www.mdpi.com/1424-8247/14/10/1016 |
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| author | Andrew L. Schilb Josef H. Scheidt Amita M. Vaidya Zhanhu Sun Da Sun Sangjoon Lee Zheng-Rong Lu |
| author_facet | Andrew L. Schilb Josef H. Scheidt Amita M. Vaidya Zhanhu Sun Da Sun Sangjoon Lee Zheng-Rong Lu |
| author_sort | Andrew L. Schilb |
| collection | DOAJ |
| description | Nucleic acids are promising for a variety of therapies, such as cancer therapy and the gene therapy of genetic disorders. The therapeutic efficacy of nucleic acids is reliant on the ability of their efficient delivery to the cytosol of the target cells. Amino lipids have been developed to aid in the cytosolic delivery of nucleic acids. This work reports a new and efficient synthetic pathway for the lipid carrier, (1-aminoethyl) iminobis [<i>N</i>-(oleicylcysteinyl-1-amino-ethyl)propionamide] (ECO). The previous synthesis of the ECO was inefficient and presented poor product quality control. A solution-phase synthesis of the ECO was explored, and each intermediate product was characterized with better quality control. The ECO was synthesized with a relatively high yield and high purity. The formulations of the ECO nanoparticles were made with siRNA, miRNA, or plasmid DNA, and characterized. The transfection efficiency of the nanoparticles was evaluated in vitro over a range of N/P ratios. The nanoparticles were consistent in size with previous formulations and had primarily a positive zeta potential. The ECO/siLuc nanoparticles resulted in potent luciferase silencing with minimal cytotoxicity. The ECO/miR-200c nanoparticles mediated the efficient delivery of miR-200c into the target cells. The ECO/pCMV-GFP nanoparticles resulted in substantial GFP expression upon transfection. These results demonstrate that the solution-phase synthetic pathway produced pure ECO for the efficient intracellular delivery of nucleic acids without size limitation. |
| first_indexed | 2024-03-10T06:16:51Z |
| format | Article |
| id | doaj.art-05d85eeca6e14b68ab0a1dd323b7a976 |
| institution | Directory Open Access Journal |
| issn | 1424-8247 |
| language | English |
| last_indexed | 2024-03-10T06:16:51Z |
| publishDate | 2021-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pharmaceuticals |
| spelling | doaj.art-05d85eeca6e14b68ab0a1dd323b7a9762023-11-22T19:36:07ZengMDPI AGPharmaceuticals1424-82472021-10-011410101610.3390/ph14101016Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic AcidsAndrew L. Schilb0Josef H. Scheidt1Amita M. Vaidya2Zhanhu Sun3Da Sun4Sangjoon Lee5Zheng-Rong Lu6Department of Biomedical Engineering, School of Engineering, Case Western Reserve University, Cleveland, OH 44106, USADepartment of Biomedical Engineering, School of Engineering, Case Western Reserve University, Cleveland, OH 44106, USADepartment of Biomedical Engineering, School of Engineering, Case Western Reserve University, Cleveland, OH 44106, USADepartment of Biomedical Engineering, School of Engineering, Case Western Reserve University, Cleveland, OH 44106, USADepartment of Biomedical Engineering, School of Engineering, Case Western Reserve University, Cleveland, OH 44106, USADepartment of Biomedical Engineering, School of Engineering, Case Western Reserve University, Cleveland, OH 44106, USADepartment of Biomedical Engineering, School of Engineering, Case Western Reserve University, Cleveland, OH 44106, USANucleic acids are promising for a variety of therapies, such as cancer therapy and the gene therapy of genetic disorders. The therapeutic efficacy of nucleic acids is reliant on the ability of their efficient delivery to the cytosol of the target cells. Amino lipids have been developed to aid in the cytosolic delivery of nucleic acids. This work reports a new and efficient synthetic pathway for the lipid carrier, (1-aminoethyl) iminobis [<i>N</i>-(oleicylcysteinyl-1-amino-ethyl)propionamide] (ECO). The previous synthesis of the ECO was inefficient and presented poor product quality control. A solution-phase synthesis of the ECO was explored, and each intermediate product was characterized with better quality control. The ECO was synthesized with a relatively high yield and high purity. The formulations of the ECO nanoparticles were made with siRNA, miRNA, or plasmid DNA, and characterized. The transfection efficiency of the nanoparticles was evaluated in vitro over a range of N/P ratios. The nanoparticles were consistent in size with previous formulations and had primarily a positive zeta potential. The ECO/siLuc nanoparticles resulted in potent luciferase silencing with minimal cytotoxicity. The ECO/miR-200c nanoparticles mediated the efficient delivery of miR-200c into the target cells. The ECO/pCMV-GFP nanoparticles resulted in substantial GFP expression upon transfection. These results demonstrate that the solution-phase synthetic pathway produced pure ECO for the efficient intracellular delivery of nucleic acids without size limitation.https://www.mdpi.com/1424-8247/14/10/1016gene therapyECOmiRNAsiRNAplasmidRNA interference |
| spellingShingle | Andrew L. Schilb Josef H. Scheidt Amita M. Vaidya Zhanhu Sun Da Sun Sangjoon Lee Zheng-Rong Lu Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic Acids Pharmaceuticals gene therapy ECO miRNA siRNA plasmid RNA interference |
| title | Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic Acids |
| title_full | Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic Acids |
| title_fullStr | Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic Acids |
| title_full_unstemmed | Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic Acids |
| title_short | Optimization of Synthesis of the Amino Lipid ECO for Effective Delivery of Nucleic Acids |
| title_sort | optimization of synthesis of the amino lipid eco for effective delivery of nucleic acids |
| topic | gene therapy ECO miRNA siRNA plasmid RNA interference |
| url | https://www.mdpi.com/1424-8247/14/10/1016 |
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