Xinwei Cheng1, Qibing Liu2, Hong Li3, Chen Kang4, Yang Liu3, Tianqi Guo3, Ke Shang3, Chengyun Yan5, Guang Cheng6,7, Robert J Lee8,9. 1. Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio, 43210, USA. 2. Department of Pharmacology, Hainan Medical University, Xueyuan Road, Haikou, 571199, Hainan, China. 3. Division of Pharmaceutics, College of Pharmacy, The Ohio State University, 500 West 12th Ave., Columbus, Ohio, 43210, USA. 4. Division of Pharmacology, College of Pharmacy, The Ohio State University, Columbus, Ohio, 43210, USA. 5. First Affiliated Hospital of Jiamusi University, Jiamusi University, Jiamusi, 154007, Heilongjiang, China. 6. State Key Laboratory of Long-Acting and Targeted Drug Delivery, Nanjing, 210061, Jiangsu, China. cheng@luye.cn. 7. Nanjing Hightech Industrial Development Zone, 28 Gaoxin Road, Nanjing, 210061, Jiangsu, China. cheng@luye.cn. 8. Department of Biomedical Engineering, The Ohio State University, Columbus, Ohio, 43210, USA. lee.1339@osu.edu. 9. Division of Pharmaceutics, College of Pharmacy, The Ohio State University, 500 West 12th Ave., Columbus, Ohio, 43210, USA. lee.1339@osu.edu.
Abstract
PURPOSE: Bcl-2 is an anti-apoptotic gene that is frequently overexpressed in human cancers. G3139 is an antisense oligonucleotide against bcl-2 that has shown limited efficacy in clinical trials. Here, we report the synthesis of a new antisense oligonucleotide containing additional chemical modifications and its delivery using nanoparticles. METHODS: An oligonucleotide G3139-GAP was synthesized, which has 2'-O-methyl nucleotides at the 5' and 3' ends based on a "gapmer" design. Furthermore, G3139-GAP was incorporated into lipid nanoparticles (LNPs) composed of DOTAP/egg PC/cholesterol/Tween 80. The LNP-loaded G3139-GAP was evaluated in A549 lung cancer cells both in vitro and in a murine xenograft model for biological activity and therapeutic efficacy. RESULTS: The LNPs showed excellent colloidal and serum stability, and high encapsulation efficiency for G3139-GAP. They have a mean particle diameter and zeta potential of 134 nm and 9.59 mV, respectively. G3139-GAP-LNPs efficiently downregulated bcl-2 expression in A549 cells, as shown by 40% and 83% reduction in mRNA and protein levels, respectively. Furthermore, G3139-GAP-LNPs were shown to inhibit tumor growth, prolong survival, and downregulate tumor bcl-2 expression in an A549 murine xenograft tumor model. These data indicate that G3139-GAP-LNPs have excellent anti-tumor efficacy and warrant further evaluation.
PURPOSE:Bcl-2 is an anti-apoptotic gene that is frequently overexpressed in humancancers. G3139 is an antisense oligonucleotide against bcl-2 that has shown limited efficacy in clinical trials. Here, we report the synthesis of a new antisense oligonucleotide containing additional chemical modifications and its delivery using nanoparticles. METHODS: An oligonucleotideG3139-GAP was synthesized, which has 2'-O-methyl nucleotides at the 5' and 3' ends based on a "gapmer" design. Furthermore, G3139-GAP was incorporated into lipid nanoparticles (LNPs) composed of DOTAP/egg PC/cholesterol/Tween 80. The LNP-loaded G3139-GAP was evaluated in A549 lung cancer cells both in vitro and in a murine xenograft model for biological activity and therapeutic efficacy. RESULTS: The LNPs showed excellent colloidal and serum stability, and high encapsulation efficiency for G3139-GAP. They have a mean particle diameter and zeta potential of 134 nm and 9.59 mV, respectively. G3139-GAP-LNPs efficiently downregulated bcl-2 expression in A549 cells, as shown by 40% and 83% reduction in mRNA and protein levels, respectively. Furthermore, G3139-GAP-LNPs were shown to inhibit tumor growth, prolong survival, and downregulate tumorbcl-2 expression in an A549 murine xenograft tumor model. These data indicate that G3139-GAP-LNPs have excellent anti-tumor efficacy and warrant further evaluation.
Entities:
Keywords:
antisense oligonucleotide; bcl-2; cancer; drug delivery; lipid nanoparticles
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