title: Novel polymeric nanoparticles for intracellular delivery of peptide Cargos: antitumor efficacy of the BCL-2 conversion peptide NuBCP-9.
abstract: The preclinical development of peptidyl drugs for cancer treatment is hampered by their poor pharmacologic properties and cell penetrative capabilities in vivo. In this study, we report a nanoparticle-based formulation that overcomes these limitations, illustrating their utility in studies of the anticancer peptide NuBCP-9, which converts BCL-2 from a cell protector to a cell killer. NuBCP-9 was encapsulated in polymeric nanoparticles composed of a polyethylene glycol (PEG)-modified polylactic acid (PLA) diblock copolymer (NuBCP-9/PLA-PEG) or PEG-polypropylene glycol-PEG-modified PLA-tetrablock copolymer (NuBCP-9/PLA-PEG-PPG-PEG). We found that peptide encapsulation was enhanced by increasing the PEG chain length in the block copolymers. NuBCP-9 release from the nanoparticles was controlled by both PEG chain length and the PLA molecular weight, permitting time-release over sustained periods. Treatment of human cancer cells with these nanoparticles in vitro triggered apoptosis by NuBCP-9-mediated mechanism, with a potency similar to NuBCP-9 linked to a cell-penetrating poly-Arg peptide. Strikingly, in vivo administration of NuBCP-9/nanoparticles triggered complete regressions in the Ehrlich syngeneic mouse model of solid tumor. Our results illustrate an effective method for sustained delivery of anticancer peptides, highlighting the superior qualities of the novel PLA-PEG-PPG-PEG tetrablock copolymer formulation as a tool to target intracellular proteins.
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: Manoj
authors.initials: M
authors.lastname: Kumar
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: Dikshi
authors.initials: D
authors.lastname: Gupta
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: Gurpal
authors.initials: G
authors.lastname: Singh
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: Sapna
authors.initials: S
authors.lastname: Sharma
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: Madhusudan
authors.initials: M
authors.lastname: Bhat
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: C K
authors.initials: CK
authors.lastname: Prashant
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: A K
authors.initials: AK
authors.lastname: Dinda
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts.
authors.forename: Surender
authors.initials: S
authors.lastname: Kharbanda
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts donald_kufe@dfci.harvard.edu harpal2000@yahoo.com.
authors.forename: Donald
authors.initials: D
authors.lastname: Kufe
authors.affiliation: Authors' Affiliations: Center for Biomedical Engineering, Indian Institute of Technology, Hauz Khas; Department of Pathology, All India Institute of Medical Sciences, Ansari Nagar, New Delhi; and Department of Medical Oncology, Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts donald_kufe@dfci.harvard.edu harpal2000@yahoo.com.
authors.forename: Harpal
authors.initials: H
authors.lastname: Singh
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D017209:Apoptosis; D002286:Carcinoma, Ehrlich Tumor / Q000188:drug therapy* / Q000473:pathology; D049109:Cell Proliferation / Q000187:drug effects; D003692:Delayed-Action Preparations / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D004354:Drug Screening Assays, Antitumor; D056945:Hep G2 Cells; D006801:Humans; D020128:Inhibitory Concentration 50; D007773:Lactates / Q000737:chemistry; D061986:MCF-7 Cells; D051379:Mice; D008807:Mice, Inbred BALB C; D053769:Nanocapsules / Q000737:chemistry*; D009368:Neoplasm Transplantation; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D010316:Particle Size; D011092:Polyethylene Glycols / Q000737:chemistry; D011409:Propylene Glycols / Q000737:chemistry; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D047368:Tumor Burden / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D003692:Delayed-Action Preparations; D007773:Lactates; D053769:Nanocapsules; D009842:Oligopeptides; D011409:Propylene Glycols; D019253:Proto-Oncogene Proteins c-bcl-2; C000590397:phenylalanyl-seryl-arginyl-seryl-leucyl-histidyl-seryl-leucyl-leucine; C088513:poly(lactic acid-ethylene glycol); D011092:Polyethylene Glycols
references.citation: Nature. 2005 Jun 2;435(7042):677-81
references.pmid: 15902208
references.citation: Cancer Res. 2009 Aug 15;69(16):6506-14
references.pmid: 19638590
references.citation: Int J Nanomedicine. 2011;6:1443-51
references.pmid: 21796246
references.citation: Adv Drug Deliv Rev. 1998 Sep 7;33(3):241-252
references.pmid: 10837664
references.citation: FASEB J. 2002 Aug;16(10):1217-26
references.pmid: 12153989
references.citation: Eur J Pharm Biopharm. 2010 Jun;75(2):96-106
references.pmid: 20211727
references.citation: Colloids Surf B Biointerfaces. 2013 Nov 1;111:282-8
references.pmid: 23838194
references.citation: Science. 2004 Jul 30;305(5684):626-9
references.pmid: 15286356
references.citation: Ann Oncol. 1994 May;5(5):409-14
references.pmid: 7915536
references.citation: Int J Nanomedicine. 2012;7:4089-98
references.pmid: 22888245
references.citation: Clin Cancer Res. 2004 Jun 1;10(11):3708-16
references.pmid: 15173077
references.citation: Clin Cancer Res. 2007 Dec 15;13(24):7264-70
references.pmid: 18094406
references.citation: Carcinogenesis. 2003 Jan;24(1):75-80
references.pmid: 12538351
references.citation: Cancer Res. 2012 Nov 1;72(21):5566-75
references.pmid: 22952218
references.citation: Chem Soc Rev. 2011 Jul;40(7):3638-55
references.pmid: 21566806
references.citation: Biomacromolecules. 2012 Apr 9;13(4):1212-22
references.pmid: 22394150
references.citation: Oncogene. 2003 Nov 24;22(53):8590-607
references.pmid: 14634621
references.citation: Cancer Cell. 2008 Oct 7;14(4):285-98
references.pmid: 18835031
references.citation: Proc Natl Acad Sci U S A. 2006 Apr 18;103(16):6315-20
references.pmid: 16606824
references.citation: J Clin Invest. 2009 Sep;119(9):2830-42
references.pmid: 19726870
references.citation: Nature. 2009 Nov 12;462(7270):182-8
references.pmid: 19907488
references.citation: J Am Assoc Lab Anim Sci. 2011 Sep;50(5):600-13
references.pmid: 22330705
references.citation: Expert Opin Drug Deliv. 2009 Oct;6(10):1033-47
references.pmid: 19637980
references.citation: Proc Natl Acad Sci U S A. 2012 Oct 30;109(44):17942-7
references.pmid: 23071338
references.citation: Mol Ther. 2011 Dec;19(12):2124-32
references.pmid: 21934653
references.citation: Curr Med Chem. 2011;18(9):1373-9
references.pmid: 21366527
references.citation: Expert Opin Drug Deliv. 2009 Oct;6(10):1049-64
references.pmid: 19743895
references.citation: Cancer Res. 2008 May 1;68(9):3421-8
references.pmid: 18451170
references.citation: Mol Pharm. 2009 May-Jun;6(3):971-7
references.pmid: 19317442
references.citation: J Control Release. 2012 Jan 30;157(2):279-86
references.pmid: 21871507
references.citation: Int J Nanomedicine. 2013;8:505-20
references.pmid: 23429428
references.citation: Curr Pharm Des. 2011;17(3):293-319
references.pmid: 21348831
references.citation: Methods Enzymol. 2008;446:387-408
references.pmid: 18603135
references.citation: Cell. 2004 Feb 20;116(4):527-40
references.pmid: 14980220
references.citation: Int J Nanomedicine. 2010 Nov 26;5:1057-65
references.pmid: 21170353
references.citation: Med Res Rev. 2007 Nov;27(6):755-95
references.pmid: 17019680
references.citation: Mol Cancer Ther. 2011 May;10(5):806-16
references.pmid: 21421804
references.citation: Nat Chem Biol. 2010 Aug;6(8):595-601
references.pmid: 20562877
references.citation: Clin Cancer Res. 2009 Jan 1;15(1):181-9
references.pmid: 19118045
references.citation: Sci Transl Med. 2012 Apr 4;4(128):128ra39
references.pmid: 22491949
references.citation: ACS Appl Mater Interfaces. 2012 May;4(5):2788-93
references.pmid: 22563747
references.citation: Cancer Cell. 2005 May;7(5):457-68
references.pmid: 15894266
references.citation: Chem Soc Rev. 2012 Apr 7;41(7):2971-3010
references.pmid: 22388185
references.citation: J Control Release. 2012 Dec 10;164(2):221-35
references.pmid: 22705248
references.citation: Annu Rev Biomed Eng. 2012;14:1-16
references.pmid: 22524388
references.citation: Proc Natl Acad Sci U S A. 2008 Nov 11;105(45):17232-7
references.pmid: 18796605
references.citation: Chem Biodivers. 2009 Jul;6(7):1101-10
references.pmid: 19623560
references.citation: Proc Natl Acad Sci U S A. 2010 Feb 2;107(5):1918-23
references.pmid: 20080680