Selection of antitumor displayed peptides for the specific delivery of the anticancer drug lactaptin

Nemudraya,Anna  Andreevna; Kuligina,Elena  Vladimirovna; Ilyichev,Alexandr  Alexeevich; Fomin,Alexandr  Sergeevich; Stepanov,Grigory  Alexandrovich; Savelyeva,Anna  Valentinovna; Koval,Olga  Alexandrovna; Richter,Vladimir  Alexandrovich

doi:10.3892/ol.2016.5266

Selection of antitumor displayed peptides for the specific delivery of the anticancer drug lactaptin

Authors:
- Anna Andreevna Nemudraya
- Elena Vladimirovna Kuligina
- Alexandr Alexeevich Ilyichev
- Alexandr Sergeevich Fomin
- Grigory Alexandrovich Stepanov
- Anna Valentinovna Savelyeva
- Olga Alexandrovna Koval
- Vladimir Alexandrovich Richter
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Affiliations: Laboratory of Biotechnology, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of The Russian Academy of Sciences, 630090 Novosibirsk, Russia, Department of Bioengineering, State Research Center of Virology and Biotechnology VECTOR, 630559 Koltsovo, Russia
Published online on: October 14, 2016 https://doi.org/10.3892/ol.2016.5266
Pages: 4547-4555
Copyright: © Nemudraya et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

It has been previously demonstrated that lactaptin, the proteolytic fragment of human milk protein κ‑casein, induces the death of various cultured cancer cells. The recombinant analog of lactaptin, RL2, effectively induces the apoptosis of mouse hepatocarcinoma‑1 (HA‑1) tumor cells in vitro and suppress the growth of HA‑1 tumors and metastases in vivo. The antitumor drug Lactaptin developed on the basis of RL2 has been successful in preclinical trials. Lactaptin shows its efficiency in relation to mouse and human cancer cells and tumors. However, Lactaptin, as with the majority of protein‑based therapeutic drugs, is distributed evenly throughout the organism, which reduces its antitumor efficacy. To develop the targeted delivery of lactaptin, the present study selected tumor‑specific peptides by screening a phage display peptide library in vivo on A/Sn strain mice with subcutaneously transplanted HA‑1 cells. Two genetic constructs were made for the production of recombinant fusion proteins composed of RL2 and the selected tumor‑targeting peptide. In vitro experiments involving HA‑1, MDA‑MB‑231 and MCF‑7 cells cultures demonstrated that the fusion proteins induce apoptotic death in mouse and human tumor cells, as with RL2. The in vivo experiments involving the mouse HA‑1 tumor model demonstrated that the tumor fluorescence intensity of the Cy5‑fusion protein conjugates is higher than that of RL2‑Cy5. As conjugation of the tumor‑specific peptides to RL2 provided retention of RL2 in the tumor tissues, fusion proteins composed of lactaptin and peptides specific for human tumors are deemed promising to improve the antitumor efficiency of lactaptin.

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1	Millimouno FM, Dong J, Yang L, Li J and Li X: Targeting apoptosis pathways in cancer and perspectives with natural compounds from mother nature. Cancer Prev Res (Phila). 7:1081–1107. 2014. View Article : Google Scholar : PubMed/NCBI
2	Nekipelaya VV, Semenov DV, Potapenko MO, Kuligina EV, Kit Yu, Romanova IV and Richter VA: Lactaptin is a human milk protein inducing apoptosis of MCF-7 adenocarcinoma cells. Dokl Biochem Biophys. 419:58–61. 2008. View Article : Google Scholar : PubMed/NCBI
3	Vlassov VV, Richter VA, Semenov DV, Nekipelaya VV, Kuligina EV and Potapenko MO: Peptide inducing apoptotic death of human cancer cells. Patent RF N 2317304. 2008.
4	Tikunova NV, Semenov DV, Babkina IN, Kuligina EV, Koval OA, et al: Recombinant plasmid DNA pFK2, providing synthesis of the recombinant peptide which is the analog of human kappa-casein and recombinant peptide-the analog of human kappa-casein fragment, with the apoptotic activity against human tumor cells. Patent RF N 2401307. 2010.
5	Semenov DV, Fomin AS, Kuligina EV, Koval OA, Matveeva VA, Babkina IN, Tikunova NV and Richter VA: Recombinant analogs of a novel milk pro-apoptotic peptide, lactaptin, and their effect on cultured human cells. Protein J. 29:174–180. 2010. View Article : Google Scholar : PubMed/NCBI
6	Koval OA, Fomin AS, Kaledin VI, Semenov DV, Potapenko MO, Kuligina EV, Nikolin VP, Nikitenko EV and Richter VA: A novel pro-apoptotic effector lactaptin inhibits tumor growth in mice models. Biochimie. 94:2467–2474. 2012. View Article : Google Scholar : PubMed/NCBI
7	Koval OA, Tkachenko AV, Fomin AS, Semenov DV, Nushtaeva AA, Kuligina EV, Zavjalov EL and Richter VA: Lactaptin induces p53-independent cell death associated with features of apoptosis and autophagy and delays growth of breast cancer cells in mouse xenografts. PLoS One. 9:e939212014. View Article : Google Scholar : PubMed/NCBI
8	Bondarenko DA, Richter VA, Kuligina EV, Koval OA, Fomin AS, et al: Toxicity studies and pharmacokinetics of Lactaptin. Russian Journal of Biopharmaceuticals. 7:40–47. 2015.
9	Nemudraya AA, Richter VA and Kuligina EV: Phage peptide libraries as a source of targeting ligands. Acta Naturae. 8:48–57. 2016.PubMed/NCBI
10	Tamura K, Dudley J, Nei M and Kumar S: MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol. 24:1596–1599. 2007. View Article : Google Scholar : PubMed/NCBI
11	Bakhshinejad B and Sadeghizadeh M: Bacteriophages as vehicles for gene delivery into mammalian cells: Prospects and problems. Expert Opin Drug Deliv. 11:1561–1574. 2014. View Article : Google Scholar : PubMed/NCBI
12	Galluzzi L, Vitale I, Abrams JM, Alnemri ES, Baehrecke EH, Blagosklonny MV, Dawson TM, Dawson VL, El-Deiry WS, Fulda S, et al: Molecular definitions of cell death subroutines: Recommendations of the Nomenclature Committee on Cell Death 2012. Cell Death Differ. 19:107–120. 2012. View Article : Google Scholar : PubMed/NCBI
13	Fomin AS, Koval' OA, Semenov DV, Potapenko MO, Kuligina EV, Kit IuIa and Rikhter VA: The Analysis of biochemical markers of MCF-7 cells apoptosis induced by recombinant analog of lactaptin. Bioorg Khim. 38:92–98. 2012.PubMed/NCBI
14	Svensen N, Walton JG and Bradley M: Peptides for cell-selective drug delivery. Trends Pharmacol Sci. 33:186–192. 2012. View Article : Google Scholar : PubMed/NCBI
15	Venditto VJ and Szoka FC Jr: Cancer nanomedicines: So many papers and so few drugs! Adv Drug Deliv Rev. 65:80–88. 2013. View Article : Google Scholar : PubMed/NCBI
16	Hamzeh-Mivehroud M, Alizadeh AA, Morris MB, Church WB and Dastmalchi S: Phage display as a technology delivering on the promise of peptide drug discovery. Drug Discov Today. 18:1144–1157. 2013. View Article : Google Scholar : PubMed/NCBI
17	Krumpe LR and Mori T: The use of phage-displayed peptide libraries to develop tumor-targeting drugs. Int J Pept Res Ther. 12:79–91. 2006. View Article : Google Scholar : PubMed/NCBI
18	Wicki A, Witzigmann D, Balasubramanian V and Huwyler J: Nanomedicine in cancer therapy: Challenges, opportunities, and clinical applications. J Control Release. 200:138–157. 2015. View Article : Google Scholar : PubMed/NCBI
19	Ruoslahti E: Specialization of tumour vasculature. Nat Rev Cancer. 2:83–90. 2002. View Article : Google Scholar : PubMed/NCBI
20	Li ZJ and Cho CH: Peptides as targeting probes against tumor vasculature for diagnosis and drug delivery. J Transl Med. 10:(Suppl 1). S12012. View Article : Google Scholar : PubMed/NCBI
21	Brown KC: Peptidic tumor targeting agents: The road from phage display peptide selections to clinical applications. Curr Pharm Des. 16:1040–1054. 2010. View Article : Google Scholar : PubMed/NCBI
22	Parks SK, Cormerais Y, Marchiq I and Pouyssegur J: Hypoxia optimises tumour growth by controlling nutrient import and acidic metabolite export. Mol Aspects Med 47–48. 3–14. 2016. View Article : Google Scholar
23	Wahba HA and El-Hadaad HA: Current approaches in treatment of triple-negative breast cancer. Cancer Biol Med. 12:106–116. 2015.PubMed/NCBI
24	Bose S: Triple-negative breast carcinoma: Morphologic and molecular subtypes. Adv Anat Pathol. 22:306–313. 2015. View Article : Google Scholar : PubMed/NCBI
25	Noren KA and Noren CJ: Construction of high-complexity combinatorial phage display peptide libraries. Methods. 23:169–178. 2001. View Article : Google Scholar : PubMed/NCBI
26	Ahn KY, Ko HK, Lee BR, Lee EJ, Lee JH, Byun Y, Kwon IC, Kim K and Lee J: Engineered protein nanoparticles for in vivo tumor detection. Biomaterials. 35:6422–6429. 2014. View Article : Google Scholar : PubMed/NCBI