Mehdi Soleymani-Goloujeh1,2, Ali Nokhodchi3, Mehri Niazi4, Saeedeh Najafi-Hajivar4, Javid Shahbazi-Mojarrad5, Nosratollah Zarghami6, Parvin Zakeri-Milani7, Ali Mohammadi8, Mohammad Karimi8, Hadi Valizadeh9. 1. a Department of Medical Nanotechnology, Faculty of Advanced Medical Sciences , Tabriz University of Medical Sciences , Tabriz , Iran. 2. b Department of Stem Cells and Developmental Biology at Cell Science Research Center , Royan Institute for Stem Cell Biology and Technology, ACECR , Tehran , Iran. 3. c Pharmaceutics Research Laboratory, School of Life Sciences , University of Sussex , Falmer , Brighton , UK. 4. d Student Research Committee, Faculty of Advanced Medical Sciences , Tabriz University of Medical Sciences , Tabriz , Iran. 5. e Biotechnology Research Center and Faculty of Pharmacy , Tabriz University of Medical Sciences , Tabriz , Iran. 6. f Department of Medical Biotechnology, Faculty of Advanced Medical Sciences , Tabriz University of Medical Sciences , Tabriz , Iran. 7. g Liver and Gastrointestinal Diseases Research Center and Faculty of Pharmacy , Tabriz University of Medical Sciences , Tabriz , Iran. 8. h Immunology Research Center , Tabriz University of Medical Sciences , Tabriz , Iran. 9. i Drug Applied Research Center and Faculty of Pharmacy , Tabriz University of Medical Sciences , Tabriz , Iran.
Abstract
PURPOSE: To assess the effect of "N-Acetylation and C-Amidation" on the cellular uptake, cytotoxicity and performance of amphiphilic cell penetrating peptides (CPP) loaded with methotrexate (MTX). METHODS: Several CPPs were synthesized by solid phase peptide synthesis method. Some of these sequences were modified with pyroglutamic acid at N-terminus and benzylamine or memantine at C-terminus. The resultant nanomaterials were prepared due to the physical linkage between CPPs and MTX. The internalization and cytotoxicity of both CPP-MTX bioconjugates and unmodified CPPs against MCF-7 human breast adenocarcinoma cells was evaluated. RESULTS: N-l and C-terminal modification did not alter the toxicity of CPPs. Physical linkage of CPPs with MTX resulted in a lower drug loading efficiency in comparison with chemically conjugated CPP-MTX bio-conjugates. Both nano-complexes increase the toxic effect of MTX on MCF-7 cells. Furthermore, N- and C-terminal modification may cause a tangible reduction in cellular uptake of CPPs. CONCLUSION: In conclusion, it was shown that cytotoxicity of modified peptides which were physically linked with MTX, considerably higher than both physically loaded unmodified peptides and chemically conjugated peptides with MTX. Also, cell internalization was reduced after peptide end-protection. These findings confirmed the effectiveness of N- and C-terminal modifications on cell viability and CPPs internalization.
PURPOSE: To assess the effect of "N-Acetylation and C-Amidation" on the cellular uptake, cytotoxicity and performance of amphiphilic cell penetrating peptides (CPP) loaded with methotrexate (MTX). METHODS: Several CPPs were synthesized by solid phase peptide synthesis method. Some of these sequences were modified with pyroglutamic acid at N-terminus and benzylamine or memantine at C-terminus. The resultant nanomaterials were prepared due to the physical linkage between CPPs and MTX. The internalization and cytotoxicity of both CPP-MTX bioconjugates and unmodified CPPs against MCF-7 humanbreast adenocarcinoma cells was evaluated. RESULTS: N-l and C-terminal modification did not alter the toxicity of CPPs. Physical linkage of CPPs with MTX resulted in a lower drug loading efficiency in comparison with chemically conjugated CPP-MTX bio-conjugates. Both nano-complexes increase the toxic effect of MTX on MCF-7 cells. Furthermore, N- and C-terminal modification may cause a tangible reduction in cellular uptake of CPPs. CONCLUSION: In conclusion, it was shown that cytotoxicity of modified peptides which were physically linked with MTX, considerably higher than both physically loaded unmodified peptides and chemically conjugated peptides with MTX. Also, cell internalization was reduced after peptide end-protection. These findings confirmed the effectiveness of N- and C-terminal modifications on cell viability and CPPs internalization.