Brevinin-2R and Derivatives as Potential Anticancer Peptides: Synthesis, Purification, Characterization and Biological Activities
- 155 Downloads
Brevinin-2R (BR-2R), a novel antimicrobial and anti-cancer peptide and two derivatives, BR-C and BR-D, were well designed, synthesized and purified by RP-HPLC (reverse phase high pressure chromatography). Accuracy was confirmed through mass spectrometry. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide) assay was carried out to determine cytotoxicity of various concentrations of peptides and their anti-proliferative potential. Results indicate that these compounds have significant anticancer activities in vitro on both MCF-7 (human breast adenocarcinoma) and A549 (human lung carcinoma) cancer cells in a concentration-dependent manner, although showed less cytotoxicity on human red blood cells. Further investigation verified that apoptosis induction, confirmed by Annexin V/PI staining and strong sub G1 cell cycle arrest, was observed in treated MCF-7 and A549 cells. Induction of apoptosis and anti-proliferating activity were closely associated with augmentation of caspase-3/7 activity, surging the apoptotic cells with no change in caspase-8 activity. The growth inhibition of BR-2R was exceedingly stronger than two derivatives; on the other hand, MCF-7 was more susceptible to treatment than A549. Findings prominently suggested development of Brevinin-2R and derivatives as novel promising compounds for cancer therapy, which may conquer drug resistance.
KeywordsAnticancer activity Apoptosis Brevinin-2R Cell cycle Chemical synthesis
This study was financially supported by Islamic Azad University of Ardabil. Authors are grateful to Dr. Hossein Akbari from department of physics at Islamic Azad University, Ardabil Branch and Mehdi Hassanvand Jamadi from Shahid Beheshti University for critically reviewing the manuscript and their invaluable contributions to data analysis.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
Informed consent was obtained from all individual participants included in the study.
- Amaral AC, Silva ON, Mundim NC, de Carvalho MJ, Migliolo L, Leite JR, Prates MV, Bocca AL, Franco OL, Felipe MS (2012) Predicting antimicrobial peptides from eukaryotic genomes: in silico strategies to develop antibiotics. Peptides 37:301–308. https://doi.org/10.1016/j.peptides.2012.07.021 CrossRefGoogle Scholar
- Brinckerhoff LH, Kalashnikov VV, Thompson LW, Yamshchikov GV, Pierce RA, Galavotti HS, Engelhard VH, Slingluff CL Jr (1999) Terminal modifications inhibit proteolytic degradation of an immunogenic MART-1(27–35) peptide: implications for peptide vaccines. Int J Cancer 83:326–334CrossRefGoogle Scholar
- Coffelt SB, Waterman RS, Florez L, Höner zu Bentrup K, Zwezdaryk KJ, Tomchuck SL, LaMarca HL, Danka ES, Morris CA, Scandurro AB (2008) Ovarian cancers overexpress the antimicrobial protein hCAP-18 and its derivative LL-37 increases ovarian cancer cell proliferation and invasion. Int J Cancer 122:1030–1039. https://doi.org/10.1002/ijc.23186 CrossRefGoogle Scholar
- Eliassen LT, Berge G, Leknessund A, Wikman M, Lindin I, Løkke C, Ponthan F, Johnsen JI, Sveinbjørnsson B, Kogner P, Flaegstad T, Rekdal Ø (2006) The antimicrobial peptide, Lactoferricin B, is cytotoxic to neuroblastoma cells in vitro and inhibits xenograft growth in vivo. Int J Cancer 119:493–500. https://doi.org/10.1002/ijc.21886 CrossRefGoogle Scholar
- Gupta SD, Debnath A, Saha A, Giri B, Tripathi G, Vedasiromoni JR, Gomes A, Gomes A (2007) Indian black scorpion (Heterometrus bengalensis Koch) venom induced antiproliferative and apoptogenic activity against human leukemic cell lines U937 and K562. Leuk Res 31:817–825. https://doi.org/10.1016/j.leukres.2006.06.004 CrossRefGoogle Scholar
- Kakde D, Jain D, Shrivastava V, Kakde R, Patil AT (2011) Cancer therapeutics-opportunities, challenges and advances in drug delivery. J Appl Pharm Sci 1:1–10Google Scholar
- Lee VH (1988) Enzymatic barriers to peptide and protein absorption. Crit Rev Ther Drug Carrier Syst 5:69–97Google Scholar
- Reddy TS, Reddy VG, Kulhari H, Shukla R, Kamal A, Bansal V (2016) Synthesis of (Z)-1-(1,3-diphenyl-1H-pyrazol-4-yl)-3-(phenylamino) prop-2-en-1-one derivatives as potential anticancer and apoptosis inducing agents. Eur J Med Chem 117:157–166. https://doi.org/10.1016/j.ejmech.2016.03.051 CrossRefGoogle Scholar
- Satitmanwiwat S, Changsangfa C, Khanuengthong A, Promthep K, Roytrakul S, Arpornsuwan T, Saikhun K, Sritanaudomchai H (2016) The scorpion venom peptide BmKn2 induces apoptosis in cancerous but not in normal human oral cells. Biomed Pharmacother 84:1042–1050. https://doi.org/10.1016/j.biopha.2016.10.041 CrossRefGoogle Scholar
- Sotomayor S, Muñoz-Moreno L, Carmena MJ, Schally AV, Sánchez-Chapado M, Prieto JC, Bajo AM (2010) Regulation of HER expression and transactivation in human prostate cancer cells by atargeted cytotoxic bombesin analog (AN-215) and a bombesin antagonist (RC-3095). Int J Cancer 127:1813–1822. https://doi.org/10.1002/ijc.25192 CrossRefGoogle Scholar
- Thakur R, Kini S, Kurkalang S, Banerjee A, Chatterjee P, Chanda A, Chatterjee A, Panda D, Mukherjee AK (2016) Mechanism of apoptosis induction in human breast cancer MCF-7 cell by Ruviprase, a small peptide from Daboia russelii russelii venom. Chem Biol Interact 258:297–304. https://doi.org/10.1016/j.cbi.2016.09.004 CrossRefGoogle Scholar
- Van Brussel JP, Mickisch GH (2003) Multidrug resistance in prostate cancer. Onkologie 26:175–181Google Scholar