title: Design and In Vitro Evaluation of Bispecific Complexes and Drug Conjugates of Anticancer Peptide, LyP-1 in Human Breast Cancer.
abstract: LyP-1, a nine-amino-acid tumor homing peptide, selectively binds to its cognate receptor, p32. Overexpression of p32 in certain tumors should allow use of LyP-1 as a targeting agent for the delivery of therapeutic or diagnostic agents. Peptide conjugates are developed for enhanced pre-targeting of MDA-MB-231 breast cancer cells with peptide-antibody bispecific complexes and targeting with multiple-drug/-fluorophore-conjugated nano-polymers.
  
  
  
  LyP-1-anti-DTPA bispecific antibody complexes (LyP-1-bsAbCx) were generated by conjugation of anti-DTPA antibody and LyP-1. LyP-1-doxorubicin (Dox), Dox-DTPA-succinyl-polylysine (Dox-DSPL), Dox-DSPL-LyP-1, DTPA-Dox-poly glutamic acid (D-Dox-PGA) or DTPA-rhodamine conjugated polylysine (DSPL-RITC) were prepared. In vitro therapeutic efficacy and targeting by immunofluorescence in MDA-MB-231 breast cancer cells were assessed with Dox-LyP-1. Immunofluorescence visualization of cancer cells was evaluated after pretargeting with LyP-1-bsAbCx and targeting with DSPL-RITC.
  
  
  
  Cytotoxicity of Dox-LyP-1 conjugates was significantly greater than free doxorubicin (p < 0.0001). For fluorescent-labeled LyP-1, internalization occurred in 30 min in tumor cells. Fluorescence intensity of two-step targeted cells showed that pretargeting with LyP-1-bsAbC, followed by targeting with DSPL-RITC was greater than non-pretargeted DSPL-RITC (p < 0.05).
  
  
  
  Peptide-conjugates are effective targeting agents for MDA-MB-231 breast cancer cells in culture. LyP-1-bsAbCx and Dox-LyP-1 conjugates may allow development of novel targeted cancer therapy and diagnosis.
authors.affiliation: Pharmaceutical Technology Department, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey.
authors.forename: Selin Seda
authors.initials: SS
authors.lastname: Timur
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, Bouve College of Health Sciences, Northeastern University, 360 Huntington Ave, Boston, Massachusetts, USA.
authors.forename: Prashant
authors.initials: P
authors.lastname: Bhattarai
authors.affiliation: Pharmaceutical Technology Department, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey.
authors.forename: Reyhan Neslihan
authors.initials: RN
authors.lastname: Gürsoy
authors.affiliation: Pharmaceutical Technology Department, Faculty of Pharmacy, Hacettepe University, Ankara, Turkey.
authors.forename: İmran
authors.initials: İ
authors.lastname: Vural
authors.affiliation: Department of Pharmaceutical Sciences, School of Pharmacy, Bouve College of Health Sciences, Northeastern University, 360 Huntington Ave, Boston, Massachusetts, USA. b.khaw@neu.edu.
authors.forename: Ban-An
authors.initials: BA
authors.lastname: Khaw
mesh_terms: D018033:Antibodies, Bispecific / Q000008:administration & dosage* / Q000737:chemistry*; D000970:Antineoplastic Agents / Q000008:administration & dosage* / Q000737:chemistry*; D001943:Breast Neoplasms / Q000188:drug therapy*; D045744:Cell Line, Tumor; D004317:Doxorubicin / Q000008:administration & dosage / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry; D016503:Drug Delivery Systems / Q000379:methods; D005260:Female; D006801:Humans; D053758:Nanoparticles / Q000008:administration & dosage / Q000737:chemistry; D004369:Pentetic Acid / Q000008:administration & dosage / Q000737:chemistry; D010456:Peptides, Cyclic / Q000008:administration & dosage* / Q000737:chemistry*; D011107:Polylysine / Q000008:administration & dosage / Q000031:analogs & derivatives / Q000737:chemistry; D011108:Polymers / Q000737:chemistry
chemical_list: D018033:Antibodies, Bispecific; D000970:Antineoplastic Agents; D004337:Drug Carriers; C533969:LyP-1 peptide; D010456:Peptides, Cyclic; D011108:Polymers; C060309:succinyl-polylysine; D011107:Polylysine; D004369:Pentetic Acid; D004317:Doxorubicin
keywords: peptide-antibody bispecific complexes; polymer pro-drug conjugates; pre-targeted drug targeting