title: Characterization and evaluation of amine-modified graphene amphotericin B for the treatment of visceral leishmaniasis: in vivo and in vitro studies.
abstract: Amphotericin B (AmB) has been the first-line treatment for visceral leishmaniasis (VL), a neglected protozoan disease, especially in regions like Bihar, India, where resistance to antimonials is widespread. However, adverse drug reactions are a major limiting factor. We evaluated a novel formulation of AmB conjugated to amine-modified graphene (f-Gr) for safety and efficacy over conventional AmB. The f-Gr was prepared in a gentle one-step process of noncovalent (amine) functionalization with the help of amino acid L-cysteine. This f-Gr was further conjugated to AmB by peptide bond. The conjugate (f-Gr-AmB) was characterized by Raman spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, and transmission electron microscopy. f-Gr-AmB was found to exhibit lesser cytotoxicity toward J774A.1 cells than AmB, and did not induce any hepatic or renal toxicity in Swiss albino mice. In vitro antileishmanial assay in J774A.1 cells showed significantly enhanced efficacy of f-Gr-AmB over AmB. Furthermore, percentage inhibition of amastigote replication in a hamster model of VL was significantly higher in the f-Gr-AmB treated group (87.8%) compared to the AmB group (70.4%). These results suggest that f-Gr-AmB could be a safe and effective alternative to conventional AmB in the treatment of VL.
authors.affiliation: Infectious Disease Research Laboratory, Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.
authors.forename: Shyam Lal
authors.initials: SL
authors.lastname: Mudavath
authors.affiliation: Nanoscience Unit, Department of Physics, Banaras Hindu University, Varanasi, India.
authors.forename: Mahe
authors.initials: M
authors.lastname: Talat
authors.affiliation: Infectious Disease Research Laboratory, Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.
authors.forename: Madhukar
authors.initials: M
authors.lastname: Rai
authors.affiliation: Nanoscience Unit, Department of Physics, Banaras Hindu University, Varanasi, India.
authors.forename: Onkar Nath
authors.initials: ON
authors.lastname: Srivastava
authors.affiliation: Infectious Disease Research Laboratory, Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India.
authors.forename: Shyam
authors.initials: S
authors.lastname: Sundar
mesh_terms: D000588:Amines / Q000008:administration & dosage / Q000737:chemistry*; D000666:Amphotericin B / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D000981:Antiprotozoal Agents / Q000008:administration & dosage / Q000737:chemistry* / Q000494:pharmacology / Q000627:therapeutic use*; D002460:Cell Line; D006108:Graphite / Q000008:administration & dosage / Q000737:chemistry*; D007893:Leishmania donovani / Q000187:drug effects*; D007898:Leishmaniasis, Visceral / Q000188:drug therapy* / Q000469:parasitology; D008297:Male; D008647:Mesocricetus; D051379:Mice; D008968:Molecular Conformation; D021261:Parasitic Sensitivity Tests; D017550:Spectroscopy, Fourier Transform Infrared
chemical_list: D000588:Amines; D000981:Antiprotozoal Agents; D006108:Graphite; D000666:Amphotericin B
keywords: Raman spectroscopy; amastigote; antileishmanial; cytotoxicity; efficacy; macrophage