- Email: [email protected]
Nanobody-functionalized polymersomes
Abstracts / Journal of Controlled Release 213 (2015) e8–e152
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References [1] C.C. Lee, E.R. Gillies, M.E. Fox, S.J. Guillaudeu, J.M.J. Frechet, E.E. Dy, F.C. Szoka, A single dose of doxorubicin-functionalized bowtie dendrimer cures mice bearing C-26 colon carcinomas, Proc. Natl. Acad. Sci. 103 (2006) 16649–16654. [2] A. Badros, O. Goloubeva, J.S. Dalal, I. Can, J. Thompson, A.P. Rapoport, M. Heyman, G. Akpek, R.G. Fenton, Neurotoxicity of bortezomib therapy in multiple myeloma: a single-center experience and review of the literature, Cancer 110 (2007) 1042–1049. [3] J. Su, F. Chen, V.L. Cryns, P.B. Messersmith, Catechol polymers for pH-responsive, targeted drug delivery to cancer cells, J. Am. Chem. Soc. 133 (2011) 11850–11853.
cancer cell lines including BCap-37, MDA-MB-231, MCF-7 and MCF-7 ADR treated by different drugs. While metformin only slightly reduced the proportion of CSCs in the cell lines, the treatment with PGMA– PMPC-BG substantially decreased the proportion of CSCs (CD44+/ CD24−) in all the cell lines. For instance, the CSC proportion of BCap37 cells decreased from 9% to less than 1% (Fig. 1). The drug-resistant strain MCF-7 ADR is abundant of CSCs up to 94%, but the population of CSCs dropped to 35% after treatment with PGMA–PMPC-BG. In summary, we successfully synthesized a macromolecular biguanidine drug showing remarkably enhanced anti-CSC activity in vitro, which showed great potential in improving anticancer therapy by overcoming the multidrug resistance and inhibiting the metastasis of tumors.
doi:10.1016/j.jconrel.2015.05.130
Keywords: biguanidine, macromolecular drug, cancer stem cell, chemotherapy
In vitro inhibition of cancer stem cells by biguanidine-based macromolecular drug
References [1] J.E. Visvader, G.J. Lindeman, Cancer stem cells in solid tumours: accumulating evidence and unresolved questions, Nat. Rev. Cancer 8 (2008) 755–768. [2] H.A. Hirsch, D. Iliopoulos, P.N. Tsichlis, K. Struhl, Metformin selectively targets cancer stem cells, and acts together with chemotherapy to block tumor growth and prolong remission, Cancer Res. 69 (2009) 7507–7511.
Mingzhou Ye, Jianbin Tang*, Meihua Sui, Xiangrui Liu, Youqing Shen Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Center for Bionanoengineering, Department of Chemical and Biological Engineering, Zhejiang University, Zhejiang 310027, China ⁎Corresponding author. E-mail address: [email protected] (J. Tang).
doi:10.1016/j.jconrel.2015.05.131 Cancer stem cell (CSC) is a rare population in tumor cells that have tumor-forming, self-renewal ability and differentiation capacity [1]. In recent years, increasing evidence has been obtained that CSCs have crucial effects on the formation, development and metastasis, as well as multi-drug resistance of tumors. Thus, a method that could selectively eliminate CSCs is significantly important in cancer treatment. Recently, biguanidine structures like metformin, which is a commonly used diabetes drug, were discovered to have specific effects on CSCs [2]. In this study, we conjugated biguanidine groups onto polymer chains to form a macromolecular biguanidine drug. The anti-CSC effect of the structure was studied in vitro in comparison with the small molecule metformin, to verify whether it can be a good candidate for CSC inhibition. For good solubility, biodegradability and prolonged circulation time in blood, poly(glycidyl methacrylate)–poly(2-methacryloyloxy)ethyl phosphorylcholine (PGMA–PMPC) copolymer, which was synthesized by ATRP of GMA and MPC in methanol, was chosen as a carrier for biguanidine. The macromolecular biguanidine drug, PGMA–PMPC-BG, was synthesized through reaction between thiol-functionalized biguanidine and the epoxy groups in PGMA–PMPC. All the products were well characterized by 1H NMR. To evaluate the specific effect of PGMA–PMPC-BG on CSC, flow cytometry and fluorescently-labeled antibodies are employed to monitor the CSC proportion in different
Fig. 1. Cancer stem cell (CSC) proportions in BCap-37 cell lines treated with different drugs measured by FACS.
Nanobody-functionalized polymersomes Tao Zoua,1, Fatimata Dembelea,1, Anne Beugnetb, Lucie Sengmanivongc, Ario de Marcob, Min-Hui Lia,* a Institut Curie-CNRS-Université Pierre & Marie Curie, Laboratoire PhysicoChimie Curie, UMR168, 26 Rue d'Ulm, 75248 Paris, France b Therapeutic Antibody Platform, Institut Curie, 3-5 Impasse Reille, 75014 Paris, France c Cell and Tissue Imaging Core Facility (PICT-IBiSA) and Nikon Imaging Centre@Institut Curie, UMR144, Institut Curie, Centre de Recherche, Paris, France ⁎Corresponding author. E-mail address: [email protected] (M.-H. Li). 1 Tao Zou and Fatimata Dembele have contributed equally to the work presented in this poster. As drug carriers, polymersomes (Ps) have many advantages over the classical liposomes, including adequate stability, long circulation in bloodstream, good mechanical properties, and great potential for advanced chemical functionalization [1]. Targeting properties can be endowed to the Ps by the grafting of biologically relevant receptorspecific ligands. This work focused on the development of targeted Ps. Antibody fragments derived from the heavy chain-only antibodies of Camelidae (called nanobodies or VHHs) are used as targeting ligands [2]. Advantages of using VHHs instead of conventional antibodies for surface functionalization of nanocarriers are numerous. Nanobodies can be produced cost-efficiently in bacterial and yeast expression systems; they are more stable under physiologic conditions; because of their efficiently refolding after denaturation, they are compatible with harsh functionalization procedures; since they have a mass of only 14 kDa but binding affinities comparable to those of conventional antibodies, they can be accommodated at high density on the surface of Ps. However, to the best of our knowledge, only one example of nanobody-functionalized Ps, based on poly(styrene)-block-poly(ethylene glycol), has been published [3]. In the present study, we prepared and characterized Ps made of biocompatible, biodegradable and FDA-approved poly (ethylene glycol)-block-poly (ε-caprolactone) (PEG-b-PCL) functionalized with
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Abstracts / Journal of Controlled Release 213 (2015) e8–e152
anti-HER2 VHHs (VHH1) using cysteine–maleimide coupling. The results of cell flow cytometry and uptake experiments indicated that these VHH1-functionalized Ps targeted specifically breast cancer cells expressing HER2 comparing with anti-IL2 VHHs functionalized Ps. This study will be extended in the future to other Ps, especially stimuli-responsive Ps, for targeted and controlled drug delivery.
temperature responsive poly(N-isopropylacrylamide) (p(NIPAM)) with a biocompatible hyperbranched polyglycidol (HBP) [1]. A series of novel HBP-b-[p(NIPAM)n]6 (n = 15, 25, 35, 45) chimeric copolymers were successfully synthesized by combining ring-opening multi branching polymerization of glycidol, followed by nitroxide-mediated polymerization by utilizing HBP-TEMPO as macroinitiator. For the fabrication of micelles, the polymers were dissolved in water and the solution temperature was increased above lower critical solution temperature, resulting in fairly uniform nano-sized micelles with low cytotoxicity to the normal cell lines. Doxorubicin (Dox) could be loaded in the micelles simply by adding Dox during the self-assembly procedure [2]. The highly uniform micelles show excellent Dox loading efficiency (34.6%–39.1%) and the Dox loaded micelles show temperature-dependent drug delivery in phosphate buffer saline (Fig. 1). Thus, the results suggest that HBP-b[p(NIPAM)n]6 micelles are a promising temperature responsive vehicles to deliver Dox for improved cancer therapy at physiological conditions.
= PCL-b-PEG = Fluorescent dye = Nanobody
(a)
(b)
Fig. 1. (a) Scheme of nanobody-functionalized Ps and (b) confocal micrograph of SKBR3 cells (blue: DAPI-stained nucleus) incubated 2 h at 4 °C with FITC- and VHH1functionalized Ps (green). Fig. 1. Conceptual diagram of Dox delivery of HBP-b-[p(NIPAM)n]6.
Keywords: polymersome, nanobody, targeting, PEG-b-PCL References [1] F. Meng, Z. Zhong, J. Feijen, Stimuli-responsive polymersomes for programmed drug delivery, Biomacromolecules 10 (2009) 197–209. [2] A.d. Marco, Perspectives offered by single-domain antibodies in clinical diagnostic of pediatric tumors, Curr. Med. Chem. (2013) 2188–2194. [3] M.F. Debets, W.P.J. Leenders, K. Verrijp, M. Zonjee, S.A. Meeuwissen, I. Otte-Höller, J.C.M. van Hest, Nanobodyfunctionalized polymersomes for tumor-vessel targeting, Macromol. Biosci. 13 (2013) 938–945. doi:10.1016/j.jconrel.2015.05.132
Synthesis of hyperbranched polyglycidol-b-poly(N-isopropylacrylamide) using nitroxide-mediated polymerization for thermo-sensitive drug delivery system Min Seon Heoa, Eun Jin Seob, Johnson V. Johna, Il Ho Jangb, Jae Ho Kimb, Il Kima,* a BK21 PLUS Center for Advanced Chemical Technology, Department of Polymer Science and Engineering, Pusan National University, Pusan 609735, Republic of Korea b Department of Physiology, Pusan National University School of Medicine, Yangsan 626-870, Republic of Korea ⁎Corresponding author. E-mail addresses: [email protected] (M.S. Heo), [email protected] (I. Kim). To date, the creative design of polymeric nanocarriers possessing thermo responsive behavior attracts substantial interest in the fabrication of smart materials for drug delivery systems. In this aspect, we have focused on a new multi-arm hyperbranched copolymer combining a
Keywords: hyperbranched polyglycidol, nitroxide-mediated polymerization, thermo sensitivity, drug delivery system Acknowledgments This work was supported by the Fusion Research Program for Green Technologies through the National Research Foundation of Korea (2012M3C1A1054502) and BK21 PLUS Program. References [1] J. Wang, M.H. Kim, D.E. Kang, H. Suh, I. Kim, Synthesis of pentablock and multibranched copolymers bearing poly(ethylene glycol), hyperbranched polyglycidol, and poly(L-lactide) with biocompatibility for controlled drug release, J. Polym. Sci. Polym. Chem. 50 (2012) 2553–2564. [2] R.P. Johnson, Y.-I. Jeong, J.V. John, C.W. Chung, D.H. Kang, M. Selvaraj, H. Suh, I. Kim, Dual stimuli-responsive poly(N-isopropylacrylamide)b-poly(L-histidine) chimeric materials for the controlled delivery of doxorubicin into liver carcinoma, Biomacromolecules 14 (2013) 1434–1443. doi:10.1016/j.jconrel.2015.05.133
Self-aggregates of hyperbranched epoxidized 2-hydroxyethyl methacrylate conjugates of methotrexate: Synthesis and in vitro drug delivery Xinhua Huanga,b, Mi Ri Kima, Hye Ri Leea, Min Seon Heoa, Il Kima,* a BK21 PLUS Center for Advanced Chemical Technology, Department of Polymer Science and Engineering, Pusan National University, Pusan 609-735, Republic of Korea b School of Materials Science and Engineering, Anhui University of Science and Technology, Huainan 232001, China
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References [1] C.C. Lee, E.R. Gillies, M.E. Fox, S.J. Guillaudeu, J.M.J. Frechet, E.E. Dy, F.C. Szoka, A single dose of doxorubicin-functionalized bowtie dendrimer cures mice bearing C-26 colon carcinomas, Proc. Natl. Acad. Sci. 103 (2006) 16649–16654. [2] A. Badros, O. Goloubeva, J.S. Dalal, I. Can, J. Thompson, A.P. Rapoport, M. Heyman, G. Akpek, R.G. Fenton, Neurotoxicity of bortezomib therapy in multiple myeloma: a single-center experience and review of the literature, Cancer 110 (2007) 1042–1049. [3] J. Su, F. Chen, V.L. Cryns, P.B. Messersmith, Catechol polymers for pH-responsive, targeted drug delivery to cancer cells, J. Am. Chem. Soc. 133 (2011) 11850–11853.
cancer cell lines including BCap-37, MDA-MB-231, MCF-7 and MCF-7 ADR treated by different drugs. While metformin only slightly reduced the proportion of CSCs in the cell lines, the treatment with PGMA– PMPC-BG substantially decreased the proportion of CSCs (CD44+/ CD24−) in all the cell lines. For instance, the CSC proportion of BCap37 cells decreased from 9% to less than 1% (Fig. 1). The drug-resistant strain MCF-7 ADR is abundant of CSCs up to 94%, but the population of CSCs dropped to 35% after treatment with PGMA–PMPC-BG. In summary, we successfully synthesized a macromolecular biguanidine drug showing remarkably enhanced anti-CSC activity in vitro, which showed great potential in improving anticancer therapy by overcoming the multidrug resistance and inhibiting the metastasis of tumors.
doi:10.1016/j.jconrel.2015.05.130
Keywords: biguanidine, macromolecular drug, cancer stem cell, chemotherapy
In vitro inhibition of cancer stem cells by biguanidine-based macromolecular drug
References [1] J.E. Visvader, G.J. Lindeman, Cancer stem cells in solid tumours: accumulating evidence and unresolved questions, Nat. Rev. Cancer 8 (2008) 755–768. [2] H.A. Hirsch, D. Iliopoulos, P.N. Tsichlis, K. Struhl, Metformin selectively targets cancer stem cells, and acts together with chemotherapy to block tumor growth and prolong remission, Cancer Res. 69 (2009) 7507–7511.
Mingzhou Ye, Jianbin Tang*, Meihua Sui, Xiangrui Liu, Youqing Shen Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Center for Bionanoengineering, Department of Chemical and Biological Engineering, Zhejiang University, Zhejiang 310027, China ⁎Corresponding author. E-mail address: [email protected] (J. Tang).
doi:10.1016/j.jconrel.2015.05.131 Cancer stem cell (CSC) is a rare population in tumor cells that have tumor-forming, self-renewal ability and differentiation capacity [1]. In recent years, increasing evidence has been obtained that CSCs have crucial effects on the formation, development and metastasis, as well as multi-drug resistance of tumors. Thus, a method that could selectively eliminate CSCs is significantly important in cancer treatment. Recently, biguanidine structures like metformin, which is a commonly used diabetes drug, were discovered to have specific effects on CSCs [2]. In this study, we conjugated biguanidine groups onto polymer chains to form a macromolecular biguanidine drug. The anti-CSC effect of the structure was studied in vitro in comparison with the small molecule metformin, to verify whether it can be a good candidate for CSC inhibition. For good solubility, biodegradability and prolonged circulation time in blood, poly(glycidyl methacrylate)–poly(2-methacryloyloxy)ethyl phosphorylcholine (PGMA–PMPC) copolymer, which was synthesized by ATRP of GMA and MPC in methanol, was chosen as a carrier for biguanidine. The macromolecular biguanidine drug, PGMA–PMPC-BG, was synthesized through reaction between thiol-functionalized biguanidine and the epoxy groups in PGMA–PMPC. All the products were well characterized by 1H NMR. To evaluate the specific effect of PGMA–PMPC-BG on CSC, flow cytometry and fluorescently-labeled antibodies are employed to monitor the CSC proportion in different
Fig. 1. Cancer stem cell (CSC) proportions in BCap-37 cell lines treated with different drugs measured by FACS.
Nanobody-functionalized polymersomes Tao Zoua,1, Fatimata Dembelea,1, Anne Beugnetb, Lucie Sengmanivongc, Ario de Marcob, Min-Hui Lia,* a Institut Curie-CNRS-Université Pierre & Marie Curie, Laboratoire PhysicoChimie Curie, UMR168, 26 Rue d'Ulm, 75248 Paris, France b Therapeutic Antibody Platform, Institut Curie, 3-5 Impasse Reille, 75014 Paris, France c Cell and Tissue Imaging Core Facility (PICT-IBiSA) and Nikon Imaging Centre@Institut Curie, UMR144, Institut Curie, Centre de Recherche, Paris, France ⁎Corresponding author. E-mail address: [email protected] (M.-H. Li). 1 Tao Zou and Fatimata Dembele have contributed equally to the work presented in this poster. As drug carriers, polymersomes (Ps) have many advantages over the classical liposomes, including adequate stability, long circulation in bloodstream, good mechanical properties, and great potential for advanced chemical functionalization [1]. Targeting properties can be endowed to the Ps by the grafting of biologically relevant receptorspecific ligands. This work focused on the development of targeted Ps. Antibody fragments derived from the heavy chain-only antibodies of Camelidae (called nanobodies or VHHs) are used as targeting ligands [2]. Advantages of using VHHs instead of conventional antibodies for surface functionalization of nanocarriers are numerous. Nanobodies can be produced cost-efficiently in bacterial and yeast expression systems; they are more stable under physiologic conditions; because of their efficiently refolding after denaturation, they are compatible with harsh functionalization procedures; since they have a mass of only 14 kDa but binding affinities comparable to those of conventional antibodies, they can be accommodated at high density on the surface of Ps. However, to the best of our knowledge, only one example of nanobody-functionalized Ps, based on poly(styrene)-block-poly(ethylene glycol), has been published [3]. In the present study, we prepared and characterized Ps made of biocompatible, biodegradable and FDA-approved poly (ethylene glycol)-block-poly (ε-caprolactone) (PEG-b-PCL) functionalized with
e80
Abstracts / Journal of Controlled Release 213 (2015) e8–e152
anti-HER2 VHHs (VHH1) using cysteine–maleimide coupling. The results of cell flow cytometry and uptake experiments indicated that these VHH1-functionalized Ps targeted specifically breast cancer cells expressing HER2 comparing with anti-IL2 VHHs functionalized Ps. This study will be extended in the future to other Ps, especially stimuli-responsive Ps, for targeted and controlled drug delivery.
temperature responsive poly(N-isopropylacrylamide) (p(NIPAM)) with a biocompatible hyperbranched polyglycidol (HBP) [1]. A series of novel HBP-b-[p(NIPAM)n]6 (n = 15, 25, 35, 45) chimeric copolymers were successfully synthesized by combining ring-opening multi branching polymerization of glycidol, followed by nitroxide-mediated polymerization by utilizing HBP-TEMPO as macroinitiator. For the fabrication of micelles, the polymers were dissolved in water and the solution temperature was increased above lower critical solution temperature, resulting in fairly uniform nano-sized micelles with low cytotoxicity to the normal cell lines. Doxorubicin (Dox) could be loaded in the micelles simply by adding Dox during the self-assembly procedure [2]. The highly uniform micelles show excellent Dox loading efficiency (34.6%–39.1%) and the Dox loaded micelles show temperature-dependent drug delivery in phosphate buffer saline (Fig. 1). Thus, the results suggest that HBP-b[p(NIPAM)n]6 micelles are a promising temperature responsive vehicles to deliver Dox for improved cancer therapy at physiological conditions.
= PCL-b-PEG = Fluorescent dye = Nanobody
(a)
(b)
Fig. 1. (a) Scheme of nanobody-functionalized Ps and (b) confocal micrograph of SKBR3 cells (blue: DAPI-stained nucleus) incubated 2 h at 4 °C with FITC- and VHH1functionalized Ps (green). Fig. 1. Conceptual diagram of Dox delivery of HBP-b-[p(NIPAM)n]6.
Keywords: polymersome, nanobody, targeting, PEG-b-PCL References [1] F. Meng, Z. Zhong, J. Feijen, Stimuli-responsive polymersomes for programmed drug delivery, Biomacromolecules 10 (2009) 197–209. [2] A.d. Marco, Perspectives offered by single-domain antibodies in clinical diagnostic of pediatric tumors, Curr. Med. Chem. (2013) 2188–2194. [3] M.F. Debets, W.P.J. Leenders, K. Verrijp, M. Zonjee, S.A. Meeuwissen, I. Otte-Höller, J.C.M. van Hest, Nanobodyfunctionalized polymersomes for tumor-vessel targeting, Macromol. Biosci. 13 (2013) 938–945. doi:10.1016/j.jconrel.2015.05.132
Synthesis of hyperbranched polyglycidol-b-poly(N-isopropylacrylamide) using nitroxide-mediated polymerization for thermo-sensitive drug delivery system Min Seon Heoa, Eun Jin Seob, Johnson V. Johna, Il Ho Jangb, Jae Ho Kimb, Il Kima,* a BK21 PLUS Center for Advanced Chemical Technology, Department of Polymer Science and Engineering, Pusan National University, Pusan 609735, Republic of Korea b Department of Physiology, Pusan National University School of Medicine, Yangsan 626-870, Republic of Korea ⁎Corresponding author. E-mail addresses: [email protected] (M.S. Heo), [email protected] (I. Kim). To date, the creative design of polymeric nanocarriers possessing thermo responsive behavior attracts substantial interest in the fabrication of smart materials for drug delivery systems. In this aspect, we have focused on a new multi-arm hyperbranched copolymer combining a
Keywords: hyperbranched polyglycidol, nitroxide-mediated polymerization, thermo sensitivity, drug delivery system Acknowledgments This work was supported by the Fusion Research Program for Green Technologies through the National Research Foundation of Korea (2012M3C1A1054502) and BK21 PLUS Program. References [1] J. Wang, M.H. Kim, D.E. Kang, H. Suh, I. Kim, Synthesis of pentablock and multibranched copolymers bearing poly(ethylene glycol), hyperbranched polyglycidol, and poly(L-lactide) with biocompatibility for controlled drug release, J. Polym. Sci. Polym. Chem. 50 (2012) 2553–2564. [2] R.P. Johnson, Y.-I. Jeong, J.V. John, C.W. Chung, D.H. Kang, M. Selvaraj, H. Suh, I. Kim, Dual stimuli-responsive poly(N-isopropylacrylamide)b-poly(L-histidine) chimeric materials for the controlled delivery of doxorubicin into liver carcinoma, Biomacromolecules 14 (2013) 1434–1443. doi:10.1016/j.jconrel.2015.05.133
Self-aggregates of hyperbranched epoxidized 2-hydroxyethyl methacrylate conjugates of methotrexate: Synthesis and in vitro drug delivery Xinhua Huanga,b, Mi Ri Kima, Hye Ri Leea, Min Seon Heoa, Il Kima,* a BK21 PLUS Center for Advanced Chemical Technology, Department of Polymer Science and Engineering, Pusan National University, Pusan 609-735, Republic of Korea b School of Materials Science and Engineering, Anhui University of Science and Technology, Huainan 232001, China