Table Info

Table 6.

Results for the Irwin test. Mice administered with peptide 3 at a dose of 70 mg/kg (group 1; n = 3) or 140 mg/kg (group 2; n = 3)

Item	Group 1			Group 2
Item	< 5 min	15 min	> 30 min	< 5 min	15 min	> 30 min
Autonomic
Salivation
Lachrymation
Piloerection					↑
Excessive urine
Diarrhea
Abnormal breathing					↓↓	↓↓↓
Neuromuscular
Abnormal posture	↓↓	↓↓		↓↓	↓↓	↓↓↓
Muscular tone	↓↓	↓↓		↓↓	↓↓	↓↓↓
Tremors
Seizures
Catalepsy
Straub’s tail
Exophthalmos
Motor-sensory
Touch response	↓	↓		↓↓	↓↓↓	↓↓↓
Pinna reflex
Tensile response	↓	↓		↓	↓↓	↓↓↓
Behavioral
Sedation	↑	↑↑		↑↑	↑↑	↑↑↑
Reactivity to handling	↓	↓↓		↓↓	↓↓	↓↓↓
Spontaneous activity	↓↓	↓↓		↓↓	↓↓	↓↓↓
Scratching	↑↑↑			↑↑↑
Dead

In the Irwing test, differences were observed in the evaluated items compared to the control group (injection with saline solution) (n = 2). The arrow ↑ indicated an increase in the effect and ↓ a decrease. The intensity of the effect was evaluated by the number of arrows present (for example: ↓: mild intensity; ↓↓: moderate intensity; ↓↓↓: high intensity). The colors represent the number of individuals in each group that exhibited the behavior; pink (none), deep pink (one), red (two), and deep red (three)

Supplementary materials

The supplementary figures and table for this article are available at: https://www.explorationpub.com/uploads/Article/file/100891_sup_1.pdf.

Declarations

Author contributions

KJCM: Conceptualization, Methodology, Investigation, Formal analysis, Writing—original draft, Visualization, Writing—review & editing. JERC, YVC, and LFOG: Formal analysis, Resources, Writing—original draft. CMPG: Formal analysis, Resources, Writing—original draft. ACBC: Formal analysis, Resources. RFM: Conceptualization, Supervision, Funding acquisition. JELM: Conceptualization, Methodology, Writing—review & editing. ZJRM: Conceptualization, Methodology, Resources, Writing—review & editing, Funding acquisition. JEGC: Conceptualization, Methodology, Formal analysis, Resources, Writing—original draft, Visualization, Funding acquisition. All authors read and approved the submitted version.

Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

For hemolytic activity (code 05-2019) and Irwin test (code 03-2018) were approved by the institutional ethics committee of the National University of Colombia. Zebrafish experiments were approved by the Institutional Committee for Animal Care and Use of the University of the Andes, code 24-01-2020.

Consent to participate

Informed consent to participate in the study was obtained from all participants.

Consent to publication

Not applicable.

Availability of data and materials

The raw data supporting the conclusions of this manuscript will be made available by the authors, without undue reservation, to any qualified researcher.

Funding

This research was conducted with the financial support of MinCiencias with the projects: “Obtención de un prototipo peptídico promisorio para el desarrollo de un medicamento de amplio espectro para el tratamiento del cáncer de colon, cuello uterino y próstata”. Contract RC No. [845-2019]. “Desarrollo de un medicamento contra el cáncer de mama basado en un péptido polivalente derivado de la LfcinB: Estudio de la fase preclínica (fase cero), caracterización fisicoquímica de un lote del fármaco para estudios preclínicos”. RC No. [706-2018]. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Copyright

Publisher’s note

Open Exploration maintains a neutral stance on jurisdictional claims in published institutional affiliations and maps. All opinions expressed in this article are the personal views of the author(s) and do not represent the stance of the editorial team or the publisher.

References

Vasan N, Baselga J, Hyman DM. A view on drug resistance in cancer. Nature. 2019;575:299–309. [DOI] [PubMed] [PMC]

Bray F, Laversanne M, Weiderpass E, Soerjomataram I. The ever-increasing importance of cancer as a leading cause of premature death worldwide. Cancer. 2021;127:3029–30. [DOI] [PubMed]

Siegel RL, Miller KD, Fuchs HE, Jemal A. Cancer statistics, 2022. CA Cancer J Clin. 2022;72:7–33. [DOI] [PubMed]

Kaltenmeier C, Shen C, Medich DS, Geller DA, Bartlett DL, Tsung A, et al. Time to Surgery and Colon Cancer Survival in the United States. Ann Surg. 2021;274:1025–31. [DOI] [PubMed]

Benson AB, Venook AP, Al-Hawary MM, Arain MA, Chen Y, Ciombor KK, et al. Colon Cancer, Version 2.2021, NCCN Clinical Practice Guidelines in Oncology. J Natl Compr Canc Netw. 2021;19:329–59. [DOI] [PubMed]

Das S, Ciombor KK, Haraldsdottir S, Goldberg RM. Promising New Agents for Colorectal Cancer. Curr Treat Options Oncol. 2018;19:29. [DOI] [PubMed] [PMC]

Felício MR, Silva ON, Gonçalves S, Santos NC, Franco OL. Peptides with Dual Antimicrobial and Anticancer Activities. Front Chem. 2017;5:5. [DOI] [PubMed] [PMC]

Xie M, Liu D, Yang Y. Anti-cancer peptides: classification, mechanism of action, reconstruction and modification. Open Biol. 2020;10:200004. [DOI] [PubMed] [PMC]

Chauhan S, Dhawan DK, Saini A, Preet S. Antimicrobial peptides against colorectal cancer-a focused review. Pharmacol Res. 2021;167:105529. [DOI] [PubMed]

10.

Haug BE, Camilio KA, Eliassen LT, Stensen W, Svendsen JS, Berg K, et al. Discovery of a 9-mer Cationic Peptide (LTX-315) as a Potential First in Class Oncolytic Peptide. J Med Chem. 2016;59:2918–27. [DOI] [PubMed]

11.

Chiangjong W, Chutipongtanate S, Hongeng S. Anticancer peptide: Physicochemical property, functional aspect and trend in clinical application (Review). Int J Oncol. 2020;57:678–96. [DOI] [PubMed] [PMC]

12.

Camilio KA, Rekdal O, Sveinbjörnsson B. LTX-315 (Oncopore™): A short synthetic anticancer peptide and novel immunotherapeutic agent. Oncoimmunology. 2014;3:e29181. [DOI] [PubMed] [PMC]

13.

Spicer J, Marabelle A, Baurain J, Jebsen NL, Jøssang DE, Awada A, et al. Safety, Antitumor Activity, and T-cell Responses in a Dose-Ranging Phase I Trial of the Oncolytic Peptide LTX-315 in Patients with Solid Tumors. Clin Cancer Res. 2021;27:2755–63. [DOI] [PubMed]

14.

Jiang R, Lönnerdal B. Bovine lactoferrin and lactoferricin exert antitumor activities on human colorectal cancer cells (HT-29) by activating various signaling pathways. Biochem Cell Biol. 2017;95:99–109. [DOI] [PubMed]

15.

Barragán-Cárdenas AC, Insuasty-Cepeda DS, Cárdenas-Martínez KJ, López-Meza J, Ochoa-Zarzosa A, Umaña-Pérez A, et al. LfcinB-Derived Peptides: Specific and punctual change of an amino acid in monomeric and dimeric sequences increase selective cytotoxicity in colon cancer cell lines. Arabian J Chem. 2022;15:103998. [DOI]

16.

Cárdenas-Martínez KJ, Barragán-Cárdenas AC, Rosa-Arbeláez Mdl, Parra-Giraldo CM, Ochoa-Zarzosa A, Lopez-Meza JE, et al. Evaluating the In Vitro Activity and Safety of Modified LfcinB Peptides as Potential Colon Anticancer Agents: Cell Line Studies and Insect-Based Toxicity Assessments. ACS Omega. 2023;8:37948–57. [DOI] [PubMed] [PMC]

17.

Insuasty-Cepeda DS, Barragán-Cárdenas AC, Ochoa-Zarzosa A, López-Meza JE, Fierro-Medina R, García-Castañeda JE, et al. Peptides Derived from (RRWQWRMKKLG)_2-K-Ahx Induce Selective Cellular Death in Breast Cancer Cell Lines through Apoptotic Pathway. Int J Mol Sci. 2020;21:4550. [DOI] [PubMed] [PMC]

18.

Hadianamrei R, Tomeh MA, Brown S, Wang J, Zhao X. Rationally designed short cationic α-helical peptides with selective anticancer activity. J Colloid Interface Sci. 2022;607:488–501. [DOI] [PubMed]

19.

Hristova K, Wimley WC. A look at arginine in membranes. J Membr Biol. 2011;239:49–56. [DOI] [PubMed] [PMC]

20.

Li L, Vorobyov I, Allen TW. The different interactions of lysine and arginine side chains with lipid membranes. J Phys Chem B. 2013;117:11906–20. [DOI] [PubMed] [PMC]

21.

Vazdar M, Heyda J, Mason PE, Tesei G, Allolio C, Lund M, et al. Arginine “Magic”: Guanidinium Like-Charge Ion Pairing from Aqueous Salts to Cell Penetrating Peptides. Acc Chem Res. 2018;51:1455–64. [DOI] [PubMed]

22.

Arias M, Haney EF, Hilchie AL, Corcoran JA, Hyndman ME, Hancock REW, et al. Selective anticancer activity of synthetic peptides derived from the host defence peptide tritrpticin. Biochim Biophys Acta Biomembr. 2020;1862:183228. [DOI] [PubMed]

23.

Schmidt N, Mishra A, Lai GH, Wong GCL. Arginine-rich cell-penetrating peptides. FEBS Lett. 2010;584:1806–13. [DOI] [PubMed]

24.

Svenson J, Karstad R, Flaten GE, Brandsdal B, Brandl M, Svendsen JS. Altered activity and physicochemical properties of short cationic antimicrobial peptides by incorporation of arginine analogues. Mol Pharm. 2009;6:996–1005. [DOI] [PubMed]

25.

Lu J, Xu H, Xia J, Ma J, Xu J, Li Y, et al. D- and Unnatural Amino Acid Substituted Antimicrobial Peptides With Improved Proteolytic Resistance and Their Proteolytic Degradation Characteristics. Front Microbiol. 2020;11:563030. [DOI] [PubMed] [PMC]

26.

Liu X, Cao R, Wang S, Jia J, Fei H. Amphipathicity Determines Different Cytotoxic Mechanisms of Lysine- or Arginine-Rich Cationic Hydrophobic Peptides in Cancer Cells. J Med Chem. 2016;59:5238–47. [DOI] [PubMed]

27.

Yang Q, Wang C, Lang L, Zhou Y, Wang H, Shang D. Design of potent, non-toxic anticancer peptides based on the structure of the antimicrobial peptide, temporin-1CEa. Arch Pharm Res. 2013;36:1302–10. [DOI] [PubMed]

28.

Luan X, Wu Y, Shen Y, Zhang H, Zhou Y, Chen H, et al. Cytotoxic and antitumor peptides as novel chemotherapeutics. Nat Prod Rep. 2021;38:7–17. [DOI] [PubMed] [PMC]

29.

Yang S, Shin SY, Lee CW, Kim Y, Hahm K, Kim JI. Selective cytotoxicity following Arg-to-Lys substitution in tritrpticin adopting a unique amphipathic turn structure. FEBS Lett. 2003;540:229–33. [DOI] [PubMed]

30.

Kaiser E, Colescott RL, Bossinger CD, Cook PI. Color test for detection of free terminal amino groups in the solid-phase synthesis of peptides. Anal Biochem. 1970;34:595–8. [DOI] [PubMed]

31.

Casanova YV, Guerra JAR, Pérez YAU, Castro ALL, Reina GA, Castañeda JEG, et al. Antibacterial Synthetic Peptides Derived from Bovine Lactoferricin Exhibit Cytotoxic Effect against MDA-MB-468 and MDA-MB-231 Breast Cancer Cell Lines. Molecules. 2017;22:1641. [DOI] [PubMed] [PMC]

32.

Cárdenas-Martínez KJ, Grueso-Mariaca D, Vargas-Casanova Y, Bonilla-Velásquez L, Estupiñán SM, Parra-Giraldo CM, et al. Effects of Substituting Arginine by Lysine in Bovine Lactoferricin Derived Peptides: Pursuing Production Lower Costs, Lower Hemolysis, and Sustained Antimicrobial Activity. Int J Pept Res Ther. 2021;27:1751–62. [DOI]

33.

Moya-Andérico L, Vukomanovic M, Cendra MDM, Segura-Feliu M, Gil V, Río JAD, et al. Utility of Galleria mellonella larvae for evaluating nanoparticle toxicology. Chemosphere. 2021;266:129235. [DOI] [PubMed]

34.

Bravo-Chaucanés CP, Vargas-Casanova Y, Chitiva-Chitiva LC, Ceballos-Garzon A, Modesti-Costa G, Parra-Giraldo CM. Evaluation of Anti-Candida Potential of Piper nigrum Extract in Inhibiting Growth, Yeast-Hyphal Transition, Virulent Enzymes, and Biofilm Formation. J Fungi (Basel). 2022;8:784. [DOI] [PubMed] [PMC]

35.

Insuasty-Cepeda DS, Barragán-Cárdenas AC, Ardila-Chantre N, Cárdenas-Martínez KJ, Rincón-Quiñones I, Vargas-Casanova Y, et al. Non-natural amino acids into LfcinB-derived peptides: effect in their (i) proteolytic degradation and (ii) cytotoxic activity against cancer cells. R Soc Open Sci. 2023;10:221493. [DOI] [PubMed] [PMC]

36.

Cepeda DSI, Castañeda HMP, Mayor AVR, Castañeda JEG, Villamil MM, Medina RF, et al. Synthetic Peptide Purification via Solid-Phase Extraction with Gradient Elution: A Simple, Economical, Fast, and Efficient Methodology. Molecules. 2019;24:1215. [DOI] [PubMed] [PMC]

37.

Solarte VA, Conget P, Vernot J, Rosas JE, Rivera ZJ, García JE, et al. A tetrameric peptide derived from bovine lactoferricin as a potential therapeutic tool for oral squamous cell carcinoma: A preclinical model. PLoS One. 2017;12:e0174707. [DOI] [PubMed] [PMC]

38.

Flecknell P. Replacement, reduction and refinement. ALTEX. 2002;19:73–8. [PubMed]

39.

Erhirhie EO, Ihekwereme CP, Ilodigwe EE. Advances in acute toxicity testing: strengths, weaknesses and regulatory acceptance. Interdiscip Toxicol. 2018;11:5–12. [DOI] [PubMed] [PMC]

40.

Brotzmann K, Wolterbeek A, Kroese D, Braunbeck T. Neurotoxic effects in zebrafish embryos by valproic acid and nine of its analogues: the fish-mouse connection? Arch Toxicol. 2021;95:641–57. [DOI] [PubMed] [PMC]

41.

Peptide Synthesis: What Level of Purity Do You Need [Internet]. San Francisco: Biocompare; [cited 2024 Sep 10]. Available from: https://www.biocompare.com/Editorial-Articles/117894-Peptide-Synthesis/

42.

Vaezi Z, Bortolotti A, Luca V, Perilli G, Mangoni ML, Khosravi-Far R, et al. Aggregation determines the selectivity of membrane-active anticancer and antimicrobial peptides: The case of killerFLIP. Biochim Biophys Acta Biomembr. 2020;1862:183107. [DOI] [PubMed]

43.

Lorenzon EN, Piccoli JP, Santos-Filho NA, Cilli EM. Dimerization of Antimicrobial Peptides: A Promising Strategy to Enhance Antimicrobial Peptide Activity. Protein Pept Lett. 2019;26:98–107. [DOI] [PubMed] [PMC]

44.

Rahman R, Fonseka AD, Sua S, Ahmad M, Rajendran R, Ambu S, et al. Inhibition of breast cancer xenografts in a mouse model and the induction of apoptosis in multiple breast cancer cell lines by lactoferricin B peptide. J Cell Mol Med. 2021;25:7181–9. [DOI] [PubMed] [PMC]

45.

Li H, Yang H, Li P, Wang Y, Huang G, Xing L, et al. Effect of Heat Treatment on the Antitumor Activity of Lactoferrin in Human Colon Tumor (HT29) Model. J Agric Food Chem. 2019;67:140–7. [DOI] [PubMed]

46.

Allegra E, Titball RW, Carter J, Champion OL. Galleria mellonella larvae allow the discrimination of toxic and non-toxic chemicals. Chemosphere. 2018;198:469–72. [DOI] [PubMed]

47.

Chahardehi AM, Arsad H, Lim V. Zebrafish as a Successful Animal Model for Screening Toxicity of Medicinal Plants. Plants (Basel). 2020;9:1345. [DOI] [PubMed] [PMC]

48.

Redfern WS, Dymond A, Strang I, Storey S, Grant C, Marks L, et al. The functional observational battery and modified Irwin test as global neurobehavioral assessments in the rat: Pharmacological validation data and a comparison of methods. J Pharmacol Toxicol Methods. 2019;98:106591. [DOI] [PubMed]

49.

Zhao X, Kruzel M, Ting S, Sun G, Savitz SI, Aronowski J. Optimized lactoferrin as a highly promising treatment for intracerebral hemorrhage: Pre-clinical experience. J Cereb Blood Flow Metab. 2021;41:53–66. [DOI] [PubMed] [PMC]

50.

Kumari S, Ahsan SM, Kumar JM, Kondapi AK, Rao NM. Overcoming blood brain barrier with a dual purpose Temozolomide loaded Lactoferrin nanoparticles for combating glioma (SERP-17-12433). Sci Rep. 2017;7:6602. [DOI] [PubMed] [PMC]

51.

Arcella A, Oliva MA, Staffieri S, Aalberti S, Grillea G, Madonna M, et al. In vitro and in vivo effect of human lactoferrin on glioblastoma growth. J Neurosurg. 2015;123:1026–35. [DOI] [PubMed]