- Email: [email protected]
Efficient antisense oligonucleotide delivery via non-covalent complexes of folic acid and modified polyethylenimine
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Abstracts / Journal of Controlled Release 213 (2015) e8–e152
Synergetic effect observed during development of drug-eluting biodegradable microspheres for transarterial chemoembolization (TACE) of solid tumors Yujing Wanga, Daniel G. Molinb, Nienke van den Akkerb, Leo H. Koolea,c a Department of Biomedical Engineering, Faculty of Health, Medicine & Life Sciences, Maastricht University, PO Box 616, NL 6200 MD Maastricht, The Netherlands b Department of Physiology, Faculty of Health, Medicine & Life Sciences, Maastricht University, The Netherlands c Department of Biomedical Engineering, Faculty of Engineering, Malaya University, Kuala Lumpur, Malaysia E-mail addresses: [email protected] (Y. Wang), [email protected] (D.G. Molin), [email protected] (N. van den Akker), [email protected], [email protected] (L.H. Koole). Minimally invasive therapeutic strategies provide interesting options for the treatment of solid tumors, such as hepatocellular carcinomas (HCCs). It has become obvious, however, that straightforward embolization is generally not sufficient in oncology. Embolic occlusions can trigger local neovascularization, and the ultimate result can be that the tumor grows faster and further than would have been the case without any intervention. This insight has triggered the development of drug-eluting embolic beads. These perform two functions: embolization, and controlled slow release of a drug (e.g., doxorubicin) in the vicinity of each embolic particle inside the tumor. We present here the first results of a study that on one hand follows the same principle, but on the other hand introduces two new aspects: (i) biodegradability, and (ii) preloading with two different drugs: a cytostatic and an anti-angiogenic. We set out to answer questions like: is preloading advantageous (i.e., can more drug be loaded as compared to commercial products)? Does the combined release of the drugs induce any synergetic effects? Fig. 1 shows a SEM of embolic microspheres that contain 10% cisplatin and 10% sorafenib (an anti-angiogenic anti-cancer drug) by mass. We incubated these particles in cell growth medium to test the supernatants in a cell growth assay. It was observed that the combination was much more cytotoxic than the 20% cisplatin or 20% sorafenib controls. These data point at a synergetic effect, and this will be explored further using a rabbit hepatic tumor model. Biodegradable drug-loaded embolic microspheres can be manufactured. The potential utility must be explored further. Evidently, this approach provides a plethora of possibilities in terms of drugchoice and dose optimization, as would be required for personalized therapies in oncology.
Fig. 1. SEM of embolic microspheres containing cisplatin and sorafenib (left, see text). Data on cell death, caused by the supernatant.
Keywords: tumor embolization, TACE, microspheres, drug release, biodegradable References [1] C.D. Gadaleta, G. Ranieri, Trans-arterial chemoembolization as a therapy for liver tumours, Crit. Rev. Oncol. Hematol. 80 (2011) 40–53. [2] A.J. Richardson, J.M. Laurence, V.W. Lam, Transarterial chemoembolization with irinotecan in the treatment of colorectal liver metastases: systematic review, J. Vasc. Interv. Radiol. 24 (2013) 1209–1217.
doi:10.1016/j.jconrel.2015.05.112
Efficient antisense oligonucleotide delivery via non-covalent complexes of folic acid and modified polyethylenimine Shuang Yanga, Xuewei Yanga, Jing Xiea, Robert J. Leea,b, Lesheng Tenga,* a Institute of Life Sciences, Jilin University, Changchun 130012, China b Division of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus, OH, USA ⁎Corresponding author. E-mail address: [email protected] (L. Teng). In the field of gene delivery, the development of vectors with high transfection efficiency and low toxicity is of critical importance [1]. Polyethylenimine (PEI) is a cationic polymer, which has been investigated for delivering antisense oligonucleotide into tumor cells. Low molecular weight PEI (800 Da) shows much lower cytotoxicity than PEI25 k, but much less activity in transfection. Previous studies have shown that hydrophobic modification of PEI to improve cell membrane interactions greatly improves its transfection activity [2]. Folic acid (FA) has been identified as a useful component for nanoparticles, which can be incorporated into the complexes via non-covalent complexation. Non-covalent adsorption method has been used effectively because of its convenience and efficiency. Recent studies showed synthesis of electrostatically coated (PEI)/pDNA complexes with folic acid without covalent binding. Therefore, we believe that the complexes containing hydrophobically modified PEI and FA coating are promising carriers for antisense oligonucleotides. We have synthesized oleic acid conjugated PEI (800 Da) (PEI– OA), which was coated with folic acid (FA–PEI–OA). FA–PEI–OA was then evaluated for delivery of LOR-2501, which was a 20-mer phosphorothioate antisense oligonucleotide to ribonucleotide reductase R1. Low cytotoxicity was observed with the addition of FA at the tested dosage levels than PEI complexes. Folic acid can significantly
Fig. 1. Intracellular delivery of FA/PEI–OA/Cy3–LOR-2501 complexes.
Abstracts / Journal of Controlled Release 213 (2015) e8–e152
increase PEI–OA transfection activity in different cell lines (Fig. 1). The levels of cellular uptake of FA–PEI–OA/LOR-2501 were more than three times that of PEI/LOR-2501 complexes and were not affected by the numbers of folate receptors on the cell surface. In conclusion, FA–PEI–OA is a promising agent for the delivery of antisense oligonucleotide and warrants further investigation. Keywords: gene delivery, folic Acid, polyethylenimine (PEI), oleic acid, LOR-2501 Acknowledgments This work was supported by the Jilin Province Science and Technology Development Program (20130522053JH). References [1] S.Y. Lee, M.S. Huh, S. Lee, S.J. Lee, H. Chung, J.H. Park, Y.K. Oh, K. Choi, K. Kim, I.C. Kwon, Stability and cellular uptake of polymerized siRNA (poly-siRNA)/polyethylenimine (PEI) complexes for efficient gene silencing, J. Control. Release 141 (2010) 339–346. [2] W. Guo, R.J. Lee, Efficient gene delivery via non-covalent complexes of folic acid and polyethylenimine, J. Control. Release 77 (2001) 131–138. doi:10.1016/j.jconrel.2015.05.113
Phenylboronic acid-functionalized polypeptide nanogel for glucose-responsive insulin release under physiological pH Li Zhaoa,*, Jianxun Dingb, Chunsheng Xiaob, Xiuli Zhuangb, Xuesi Chenb,* a Laboratory of Building Energy-Saving Technology Engineering, Jilin Jianzhu University, Changchun 130118, China b Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China ⁎Corresponding authors. E-mail addresses: [email protected] (L. Zhao), [email protected] (X. Chen). Biocompatible and biodegradable synthetic polypeptides have been widely studied for various biomedical applications, such as tissue engineering [1], and drug and gene delivery [2]. However, polypeptides have rarely been used as phenylboronic acid (PBA)-functionalized materials for the glucose-responsive insulin delivery system [3]. Herein, novel nanogel was prepared by the ring-opening polymerization of γ-benzyl-l-glutamate N-carboxyanhydride and γ-propargyl-lglutamate N-carboxyanhydride (PLG NCA) with amino-terminated
Scheme 1. Glucose-responsive insulin release from PBA-functionalized polypeptide nanogel.
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methoxy poly(ethylene glycol) as a macroinitiator, followed by clicking of azido-modified sugar to the PLG unit and subsequent cross-linking by adipoylamidophenylboronic acid (AAPBA). Insulin, a model drug, was loaded into the glucose-sensitive polypeptide nanogel, and the release could be triggered by the presence of glucose (Scheme 1). Furthermore, in vitro tetrazolium assay and hemolysis test suggested that these nanogels are biocompatible. These intelligent nanogels are promising for self-regulated drug delivery. Keywords: drug delivery, diabetic therapy, glucose-reponsive, nanogel, polypeptide Acknowledgments This work was financially supported by the National Natural Science Foundation of China (51303174, 51321062, 51233004, 51390484, 51273196 and 51203153) and the Scientific Development Program of Jilin Province (20140520050JH). References [1] D.L. Nettles, A. Chilkoti, L.A. Setton, Applications of elastin-like polypeptides in tissue engineering, Adv. Drug Deliv. Rev. 62 (2010) 1479–1485. [2] J. Ding, F. Shi, C. Xiao, L. Lin, L. Chen, C. He, X. Zhuang, X. Chen, One-step preparation of reduction–responsive poly(ethylene glycol)–poly (amino acid)s nanogels as efficient intracellular drug delivery platforms, Polym. Chem. 2 (2011) 2857–2864. [3] L. Zhao, J. Ding, C. Xiao, P. He, Z. Tang, X. Pang, X. Zhuang, X. Chen, Glucose-sensitive polypeptide micelles for self-regulated insulin release at physiological pH, J. Mater. Chem. 22 (2012) 12319–12328. doi:10.1016/j.jconrel.2015.05.114
Thermo-and pH dual-responsive mesoporous silica nanoparticles for controlled drug release Liang Chen, Wei Feng, Xiaojun Zhou, Zhiqi Yin, Chuanglong He* College of Chemistry, Chemical engineering and Biotechnology, Donghua University, Shanghai 201620, China ⁎Corresponding author. E-mail address: [email protected] (C. He). Mesoporous silica nanoparticles (MSNs) have recently received considerable attention as carriers for the controlled drug release. Poly(N-vinyl caprolactam) (PVCL) is a representative thermosensitive polymer with excellent biocompatibility, which does not produce poisonous small amide compounds during its hydrolysis compared with the most extensively studied poly(N-isopropyl acrylamide) [1]. Herein, we fabricated a thermo- and pH-responsive drug delivery system based on VCL and acrylic acid (AA) copolymer coated MSN via precipitation polymerization, and doxorubicin hydrochloride (DOX) was loaded for cancer therapy. The ordered mesoporous structure of MSN can be obviously observed in Fig. 1A. After polymerization, these hybrid nanoparticles become larger in size and the polymer shells are about 30 nm (Fig. 1B). The DOX release study was conducted at two different temperatures in PBS of pH 4.0 and pH 7.4. As shown in Fig. 1C, the amount of released DOX reached 70% for pH 4.0 and 28% for pH 7.4 at 37 °C, while only 40% and 20% of DOX can be released below the phase transition temperature of PVCL (25 °C), respectively. The results may be attributed to the protonation of PAA at low pH and shrink of PVCL at high temperature, which make DOX diffuse more quickly from the carrier. In summary, we have developed a dual-responsive drug delivery system based on P(VCL-co-AA) coated MSN, which could
Abstracts / Journal of Controlled Release 213 (2015) e8–e152
Synergetic effect observed during development of drug-eluting biodegradable microspheres for transarterial chemoembolization (TACE) of solid tumors Yujing Wanga, Daniel G. Molinb, Nienke van den Akkerb, Leo H. Koolea,c a Department of Biomedical Engineering, Faculty of Health, Medicine & Life Sciences, Maastricht University, PO Box 616, NL 6200 MD Maastricht, The Netherlands b Department of Physiology, Faculty of Health, Medicine & Life Sciences, Maastricht University, The Netherlands c Department of Biomedical Engineering, Faculty of Engineering, Malaya University, Kuala Lumpur, Malaysia E-mail addresses: [email protected] (Y. Wang), [email protected] (D.G. Molin), [email protected] (N. van den Akker), [email protected], [email protected] (L.H. Koole). Minimally invasive therapeutic strategies provide interesting options for the treatment of solid tumors, such as hepatocellular carcinomas (HCCs). It has become obvious, however, that straightforward embolization is generally not sufficient in oncology. Embolic occlusions can trigger local neovascularization, and the ultimate result can be that the tumor grows faster and further than would have been the case without any intervention. This insight has triggered the development of drug-eluting embolic beads. These perform two functions: embolization, and controlled slow release of a drug (e.g., doxorubicin) in the vicinity of each embolic particle inside the tumor. We present here the first results of a study that on one hand follows the same principle, but on the other hand introduces two new aspects: (i) biodegradability, and (ii) preloading with two different drugs: a cytostatic and an anti-angiogenic. We set out to answer questions like: is preloading advantageous (i.e., can more drug be loaded as compared to commercial products)? Does the combined release of the drugs induce any synergetic effects? Fig. 1 shows a SEM of embolic microspheres that contain 10% cisplatin and 10% sorafenib (an anti-angiogenic anti-cancer drug) by mass. We incubated these particles in cell growth medium to test the supernatants in a cell growth assay. It was observed that the combination was much more cytotoxic than the 20% cisplatin or 20% sorafenib controls. These data point at a synergetic effect, and this will be explored further using a rabbit hepatic tumor model. Biodegradable drug-loaded embolic microspheres can be manufactured. The potential utility must be explored further. Evidently, this approach provides a plethora of possibilities in terms of drugchoice and dose optimization, as would be required for personalized therapies in oncology.
Fig. 1. SEM of embolic microspheres containing cisplatin and sorafenib (left, see text). Data on cell death, caused by the supernatant.
Keywords: tumor embolization, TACE, microspheres, drug release, biodegradable References [1] C.D. Gadaleta, G. Ranieri, Trans-arterial chemoembolization as a therapy for liver tumours, Crit. Rev. Oncol. Hematol. 80 (2011) 40–53. [2] A.J. Richardson, J.M. Laurence, V.W. Lam, Transarterial chemoembolization with irinotecan in the treatment of colorectal liver metastases: systematic review, J. Vasc. Interv. Radiol. 24 (2013) 1209–1217.
doi:10.1016/j.jconrel.2015.05.112
Efficient antisense oligonucleotide delivery via non-covalent complexes of folic acid and modified polyethylenimine Shuang Yanga, Xuewei Yanga, Jing Xiea, Robert J. Leea,b, Lesheng Tenga,* a Institute of Life Sciences, Jilin University, Changchun 130012, China b Division of Pharmaceutics, College of Pharmacy, The Ohio State University, Columbus, OH, USA ⁎Corresponding author. E-mail address: [email protected] (L. Teng). In the field of gene delivery, the development of vectors with high transfection efficiency and low toxicity is of critical importance [1]. Polyethylenimine (PEI) is a cationic polymer, which has been investigated for delivering antisense oligonucleotide into tumor cells. Low molecular weight PEI (800 Da) shows much lower cytotoxicity than PEI25 k, but much less activity in transfection. Previous studies have shown that hydrophobic modification of PEI to improve cell membrane interactions greatly improves its transfection activity [2]. Folic acid (FA) has been identified as a useful component for nanoparticles, which can be incorporated into the complexes via non-covalent complexation. Non-covalent adsorption method has been used effectively because of its convenience and efficiency. Recent studies showed synthesis of electrostatically coated (PEI)/pDNA complexes with folic acid without covalent binding. Therefore, we believe that the complexes containing hydrophobically modified PEI and FA coating are promising carriers for antisense oligonucleotides. We have synthesized oleic acid conjugated PEI (800 Da) (PEI– OA), which was coated with folic acid (FA–PEI–OA). FA–PEI–OA was then evaluated for delivery of LOR-2501, which was a 20-mer phosphorothioate antisense oligonucleotide to ribonucleotide reductase R1. Low cytotoxicity was observed with the addition of FA at the tested dosage levels than PEI complexes. Folic acid can significantly
Fig. 1. Intracellular delivery of FA/PEI–OA/Cy3–LOR-2501 complexes.
Abstracts / Journal of Controlled Release 213 (2015) e8–e152
increase PEI–OA transfection activity in different cell lines (Fig. 1). The levels of cellular uptake of FA–PEI–OA/LOR-2501 were more than three times that of PEI/LOR-2501 complexes and were not affected by the numbers of folate receptors on the cell surface. In conclusion, FA–PEI–OA is a promising agent for the delivery of antisense oligonucleotide and warrants further investigation. Keywords: gene delivery, folic Acid, polyethylenimine (PEI), oleic acid, LOR-2501 Acknowledgments This work was supported by the Jilin Province Science and Technology Development Program (20130522053JH). References [1] S.Y. Lee, M.S. Huh, S. Lee, S.J. Lee, H. Chung, J.H. Park, Y.K. Oh, K. Choi, K. Kim, I.C. Kwon, Stability and cellular uptake of polymerized siRNA (poly-siRNA)/polyethylenimine (PEI) complexes for efficient gene silencing, J. Control. Release 141 (2010) 339–346. [2] W. Guo, R.J. Lee, Efficient gene delivery via non-covalent complexes of folic acid and polyethylenimine, J. Control. Release 77 (2001) 131–138. doi:10.1016/j.jconrel.2015.05.113
Phenylboronic acid-functionalized polypeptide nanogel for glucose-responsive insulin release under physiological pH Li Zhaoa,*, Jianxun Dingb, Chunsheng Xiaob, Xiuli Zhuangb, Xuesi Chenb,* a Laboratory of Building Energy-Saving Technology Engineering, Jilin Jianzhu University, Changchun 130118, China b Key Laboratory of Polymer Ecomaterials, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, China ⁎Corresponding authors. E-mail addresses: [email protected] (L. Zhao), [email protected] (X. Chen). Biocompatible and biodegradable synthetic polypeptides have been widely studied for various biomedical applications, such as tissue engineering [1], and drug and gene delivery [2]. However, polypeptides have rarely been used as phenylboronic acid (PBA)-functionalized materials for the glucose-responsive insulin delivery system [3]. Herein, novel nanogel was prepared by the ring-opening polymerization of γ-benzyl-l-glutamate N-carboxyanhydride and γ-propargyl-lglutamate N-carboxyanhydride (PLG NCA) with amino-terminated
Scheme 1. Glucose-responsive insulin release from PBA-functionalized polypeptide nanogel.
e69
methoxy poly(ethylene glycol) as a macroinitiator, followed by clicking of azido-modified sugar to the PLG unit and subsequent cross-linking by adipoylamidophenylboronic acid (AAPBA). Insulin, a model drug, was loaded into the glucose-sensitive polypeptide nanogel, and the release could be triggered by the presence of glucose (Scheme 1). Furthermore, in vitro tetrazolium assay and hemolysis test suggested that these nanogels are biocompatible. These intelligent nanogels are promising for self-regulated drug delivery. Keywords: drug delivery, diabetic therapy, glucose-reponsive, nanogel, polypeptide Acknowledgments This work was financially supported by the National Natural Science Foundation of China (51303174, 51321062, 51233004, 51390484, 51273196 and 51203153) and the Scientific Development Program of Jilin Province (20140520050JH). References [1] D.L. Nettles, A. Chilkoti, L.A. Setton, Applications of elastin-like polypeptides in tissue engineering, Adv. Drug Deliv. Rev. 62 (2010) 1479–1485. [2] J. Ding, F. Shi, C. Xiao, L. Lin, L. Chen, C. He, X. Zhuang, X. Chen, One-step preparation of reduction–responsive poly(ethylene glycol)–poly (amino acid)s nanogels as efficient intracellular drug delivery platforms, Polym. Chem. 2 (2011) 2857–2864. [3] L. Zhao, J. Ding, C. Xiao, P. He, Z. Tang, X. Pang, X. Zhuang, X. Chen, Glucose-sensitive polypeptide micelles for self-regulated insulin release at physiological pH, J. Mater. Chem. 22 (2012) 12319–12328. doi:10.1016/j.jconrel.2015.05.114
Thermo-and pH dual-responsive mesoporous silica nanoparticles for controlled drug release Liang Chen, Wei Feng, Xiaojun Zhou, Zhiqi Yin, Chuanglong He* College of Chemistry, Chemical engineering and Biotechnology, Donghua University, Shanghai 201620, China ⁎Corresponding author. E-mail address: [email protected] (C. He). Mesoporous silica nanoparticles (MSNs) have recently received considerable attention as carriers for the controlled drug release. Poly(N-vinyl caprolactam) (PVCL) is a representative thermosensitive polymer with excellent biocompatibility, which does not produce poisonous small amide compounds during its hydrolysis compared with the most extensively studied poly(N-isopropyl acrylamide) [1]. Herein, we fabricated a thermo- and pH-responsive drug delivery system based on VCL and acrylic acid (AA) copolymer coated MSN via precipitation polymerization, and doxorubicin hydrochloride (DOX) was loaded for cancer therapy. The ordered mesoporous structure of MSN can be obviously observed in Fig. 1A. After polymerization, these hybrid nanoparticles become larger in size and the polymer shells are about 30 nm (Fig. 1B). The DOX release study was conducted at two different temperatures in PBS of pH 4.0 and pH 7.4. As shown in Fig. 1C, the amount of released DOX reached 70% for pH 4.0 and 28% for pH 7.4 at 37 °C, while only 40% and 20% of DOX can be released below the phase transition temperature of PVCL (25 °C), respectively. The results may be attributed to the protonation of PAA at low pH and shrink of PVCL at high temperature, which make DOX diffuse more quickly from the carrier. In summary, we have developed a dual-responsive drug delivery system based on P(VCL-co-AA) coated MSN, which could