Author/Authors :
Zakeri-Milani, Parvin Liver and Gastrointestinal Diseases Research Center and Faculty of Pharmacy - Tabriz University of Medical Sciences - Tabriz, Iran , Najafi-Hajivar, Saeedeh Department of Medical Nanotechnology - Faculty of Advanced Medical Sciences - Tabriz University of Medical Sciences - Tabriz, Iran , Sarfraz, Muhammad College of Pharmacy - Al Ain University - Al Ain, UAE , Nokhodchi, Ali Pharmaceutics Research Laboratory - School of Life Sciences - University of Sussex - Falmer - Brighton BN1 9QJ, United Kingdom , Mohammadi, Hamed Department of Immunology - Immunology Research Center - School of Medicine - Tabriz University of Medical Sciences - Tabriz, Iran , Montazersaheb, Soheila Faculty of Pharmacy - Stem Cell Research Center - Tabriz University of Medical Sciences - Tabriz, Iran , Niazi, Mehri Department of Medical Nanotechnology - Faculty of Advanced Medical Sciences - Tabriz University of Medical Sciences - Tabriz, Iran , Hemmatzadeh, Maryam Department of Immunology - Immunology Research Center - School of Medicine - Tabriz University of Medical Sciences - Tabriz, Iran , Soleymani-Goloujeh, Mehdi Department of Medical Nanotechnology - Faculty of Advanced Medical Sciences - Tabriz University of Medical Sciences - Tabriz, Iran , Baradaran, Behzad Department of Immunology - Immunology Research Center - School of Medicine - Tabriz University of Medical Sciences - Tabriz, Iran , Shahbazi Mojarrad, Javid Biotechnology Research Center and Faculty of Pharmacy - Tabriz University of Medical Sciences - Tabriz, Iran , Farshbaf, Masoud Department of Medical Nanotechnology - Faculty of Advanced Medical Sciences - Tabriz University of Medical Sciences - Tabriz, Iran , Gholikhani, Tooba Drug Applied Research Center and Faculty of Pharmacy - Tabriz University of Medical Sciences - Tabriz, Iran , Valizadeh, Hadi Drug Applied Research Center and Faculty of Pharmacy - Tabriz University of Medical Sciences - Tabriz, Iran
Abstract :
Methotrexate (MTX) is one of the most effective therapeutics to treat different types of solid
tumors; however, it suffers low permeability limiting its bioavailability and cellular uptake. To
tackle this, we aim to design and fabricate different types of cell-penetrating peptides (CPPs)
to improve the intracellular uptake of MTX without causing any immunogenic response. CPPs
were synthesized by the solid-phase peptide synthesis method. Peptide-MTX conjugates were
prepared via covalent binding of peptide and drug molecule. CPPs and peptide-E8 nanoparticles
were characterized using zeta-sizer and scanning electron microscopy. Cytotoxicity of CPPs and
peptide-MTX conjugates was evaluated by MTT assay. An enzyme-linked immunosorbent assay
was employed to assess the IL-6 and TNF-α cytokine release profile. Amongst all sequences,
W4R4-MTX possessed the highest loading efficiency (97%) and drug to peptide percentage
(24.02%). The lowest loading efficiency (36%) and drug to peptide percentage (8.76%) were
seen for NGRWK-MTX conjugates. The NGRWR peptide and NGRWR-E8 nanoparticles had
acceptable size (~100 nm) with spherical and rod-like structures, respectively. The selected
CPPs and peptide-MTX conjugates did not show any cytotoxicity or immunogenicity. The
fabricated peptides are represented as promising carriers to improve the intracellular delivery
of MTX to cancer cells with low immunogenic and cytotoxic effects on normal cells.
Keywords :
Immune system , Cancer , Nanoparticles , Drug delivery , CPPs
