title: Colorectal delivery and retention of PEG-Amprenavir-Bac7 nanoconjugates--proof of concept for HIV mucosal pre-exposure prophylaxis.
abstract: Local delivery of anti-HIV drugs to the colorectal mucosa, a major site of HIV replication, and their retention within mucosal tissue would allow for a reduction in dose administered, reduced dosing frequency and minimal systemic exposure. The current report describes a mucosal pre-exposure prophylaxis (mPrEP) strategy that utilizes nanocarrier conjugates (NC) consisting of poly(ethylene glycol) (PEG), amprenavir (APV), and a cell-penetrating peptide (CPP; namely Bac7, a fragment derived from bactenecin 7). APV-PEG NCs with linear PEGs (2, 5, 10, and 30 kDa) exhibited reduced (52-21%) anti-HIV-1 protease (PR) activity as compared to free APV in an enzyme-based FRET assay. In MT-2 T cells, APV-PEG3.4 kDa-FITC (APF) anti-HIV-1 activity was significantly reduced (160-fold, IC50 = 8064 nM) due to poor cell uptake, whereas it was restored (IC50 = 78.29 nM) and similar to APV (IC50 = 50.29 nM) with the addition of Bac7 to the NC (i.e., APV-PEG3.4 kDa-Bac7, APB). Flow cytometry and confocal microscopy demonstrated Bac7-PEG3.4 kDa-FITC (BPF) uptake was two- and fourfold higher than APF in MT-2 T cells and Caco-2 intestinal epithelial cells, respectively. There was no detectable punctate fluorescence in either cell line suggesting that BPF directly enters the cytosol thus avoiding endosomal entrapment. After colorectal administration in mice, BPF mucosal concentrations were 21-fold higher than APF concentrations. BPF concentrations also remained constant for the 5 days of the study suggesting that (1) the NC's structural characteristics (i.e., the size of the PEG carrier and the presence of a CPP) significantly influenced tissue persistence, and (2) the NCs were probably lodged in the lamina propria since the average rodent colon mucosal cell turnover time is 2-3 days. These encouraging results suggest that Bac7 functionalized NCs delivered locally to the colorectal mucosa may form drug delivery depots that are capable of sustaining colorectal drug concentrations. Although the exact mechanisms for tissue persistence are unclear and will require further study, these results provide proof-of-concept feasibility for mPrEP.
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Mahta
authors.initials: M
authors.lastname: Samizadeh
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Simi
authors.initials: S
authors.lastname: Gunaseelan
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Antoinette G
authors.initials: AG
authors.lastname: Nelson
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Matthew S
authors.initials: MS
authors.lastname: Palombo
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Daniel R
authors.initials: DR
authors.lastname: Myers
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Yashveer
authors.initials: Y
authors.lastname: Singh
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Usha
authors.initials: U
authors.lastname: Ganapathi
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA.
authors.forename: Zoltan
authors.initials: Z
authors.lastname: Szekely
authors.affiliation: Department of Pharmaceutics, Ernest Mario School of Pharmacy, Rutgers, The State University of New Jersey, 160 Frelinghuysen Road, Piscataway, NJ, 08854, USA. sinko@rutgers.edu.
authors.forename: Patrick J
authors.initials: PJ
authors.lastname: Sinko
mesh_terms: D000285:Administration, Rectal; D000818:Animals; D018938:Caco-2 Cells; D002219:Carbamates / Q000008:administration & dosage*; D057846:Cell-Penetrating Peptides / Q000008:administration & dosage; D005663:Furans; D015658:HIV Infections / Q000517:prevention & control*; D016333:HIV Protease / Q000378:metabolism; D017320:HIV Protease Inhibitors / Q000008:administration & dosage*; D015497:HIV-1; D006801:Humans; D007413:Intestinal Mucosa / Q000821:virology*; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D058726:Nanoconjugates / Q000008:administration & dosage*; D010456:Peptides, Cyclic / Q000008:administration & dosage; D011092:Polyethylene Glycols / Q000008:administration & dosage; D065129:Pre-Exposure Prophylaxis / Q000379:methods*; D013449:Sulfonamides / Q000008:administration & dosage*
chemical_list: D002219:Carbamates; D057846:Cell-Penetrating Peptides; D005663:Furans; D017320:HIV Protease Inhibitors; D058726:Nanoconjugates; D010456:Peptides, Cyclic; D013449:Sulfonamides; C056454:bactenecin; D011092:Polyethylene Glycols; C095108:amprenavir; D016333:HIV Protease; C514803:p16 protease, Human immunodeficiency virus 1
keywords: Amprenavir; Bactenecin 7; Cell penetrating peptide; Darunavir; HIV; Mucosal delivery; PEG conjugation; Polymeric drug delivery systems; Pre-exposure prophylaxis