Abstract
Norcantharidin (NCTD) is the demethylated analog of cantharidin, with allegedly reduced toxicity. However, there is still limited information regarding its posology and potential risk in its use in cancer treatment. Healthy BDF1 mice were intraperitoneally administered with norcantharidin (0, 3, 6, 12, and 25 mg/kg) every 24 h for 6 days. Survivor mice were euthanized, and the brain, lungs, kidneys, spleen, and liver were procured for enzymatic and histopathological analysis in the liver and kidney. DL50 were 8.86 mg/kg for females and 11.77 mg/kg for males. The treatments with 3.0 mg/kg and 6.0 mg/kg significantly modified the phosphorylase, alanine transaminase, and γ-glutamyl transferase activities; however, an organ-specific response was detected. A significant dose-dependent decrease was observed in the kidney for ROS, while the liver had the opposite effect. Histopathological analysis revealed a significant elevation in hepatocytes’ nuclei average size and total area (3 mg/kg), as well as centrilobular vein and adjacent sinusoidal capillaries showed a significant difference. The portal triad presented a significant difference in veins and capillarity count in 6 mg/kg. Renal samples showed cortex convoluted tubules’ average size significantly augmented in both doses’ groups, and tubule count was found augmented in 6 mg/kg. These physiological effects of NCTD can be exploited as treatment strategies if able to operate in an established posology and proper testing.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
Raw data has been made available.
References
Ben Salem C, Fathallah N, Hmouda H, Bouraoui K (2011) Drug-induced hypoglycaemia: an update. Drug Saf 34(1):21–45. https://doi.org/10.2165/11538290-000000000-00000
Bjornsson TD (1979) Use of serum creatinine concentrations to determine renal function. Clin Pharmacokinet 4(3):200–222. https://doi.org/10.2165/00003088-197904030-00003
Dorn DC, Kou CA, Png KJ, Moore MA (2009) The effect of cantharidins on leukemic stem cells. Int J Cancer. https://doi.org/10.1002/ijc.24157
Dzul-Caamal R, Hernández-López A, Gonzalez-Jáuregui M, Padilla SE, Girón-Pérez MI, Vega-López A (2016) Usefulness of oxidative stress biomarkers evaluated in the snout scraping, serum and Peripheral Blood Cells of Crocodylus moreletii from Southeast Campeche for assessment of the toxic impact of PAHs, metals and total phenols. Comp Biochem Physiol A Mol Integr Physiol 200:35–46. https://doi.org/10.1016/j.cbpa.2016.05.006
Farhana A, Lappin SL (2022) Biochemistry, lactate dehydrogenase. In: StatPearls [Internet], StatPearls Publishing, Treasure Island. Available from: https://www.ncbi.nlm.nih.gov/books/NBK557536/. Accessed 8 May 2022
Friedman SB, Grota LJ, Glasgow LA (1972) Differential susceptibility of male and female mice encephalomyocarditis virus: effects of castration, adrenalectonomy, and the administration of sex hormones. Infection and Inmmunity 5(5):637–644
Hammer Ø, Harper DAT, Ryan PD (2001) PAST: Paleontological Statistics Software Package for Education and Data Analysis. Palaeontol Electron 4:9. http://palaeo-electronica.org/2001_1/past/issue1_01.htm
Harrisberg J, Deseta JC, Cohen L, Temlett J, Milne FJ (1984) Cantharidin poisoning with neurological complications. S Afr Med J 65(15):614–615
Hermes-Lima M (2004) Oxygen in biology and biochemistry: role of free radicals. In: Storey KB (ed) Functional metabolism: regulation and adaptation. Wiley-Liss Inc., Hoboken, pp 319–351
Honkanen RE (1993) Cantharidin, another natural toxin that inhibits the activity of serine/threonine protein phosphatases types 1 and 2A. FEBS Lett 330(3):283–286. https://doi.org/10.1016/0014-5793(93)80889-3
Legland D, Arganda-Carreras I, Andrey P (2016) MorphoLibJ: integrated library and plugins for mathematical morphology with ImageJ. Bioinformatics 32(22):3532–3534. https://doi.org/10.1093/bioinformatics/btw413
Li YM, Casida JE (1992) Cantharidin-binding protein: identification as protein phosphatase 2A. Proc Natl Acad Sci U S A 89(24):11867–11870. https://doi.org/10.1073/pnas.89.24.11867
Linck B, Boknik P, Knapp J, Müller FU, Neumann J, Schmitz W, Vahlensieck U (1996) Effects of cantharidin on force of contraction and phosphatase activity in nonfailing and failing human hearts. Br J Pharmacol 119(3):545–550. https://doi.org/10.1111/j.1476-5381.1996.tb15706.x
Lu K, Cao M, Mao W, Sun X, Tang J, Shen Y, Sui M (2012) Targeted acid-labile conjugates of norcantharidin for cancer therapy. J Mater Chem 22:15804. https://doi.org/10.1039/c2jm33069e
Madera-Sandoval RL, Tóvári J, Lövey J, Ranđelović I, Jiménez-Orozco A, Hernández-Chávez VG, Reyes-Maldonado E, Vega-López A (2019) Combination of pentoxifylline and α-galactosylceramide with radiotherapy promotes necro-apoptosis and leukocyte infiltration and reduces the mitosis rate in murine melanoma. Acta Histochem 121(6):680–689. https://doi.org/10.1016/j.acthis.2019.06.003
Mei L, Sang W, Cui K, Zhang Y, Chen F, Li X (2019) Norcantharidin inhibits proliferation and promotes apoptosis via c-Met/Akt/mTOR pathway in human osteosarcoma cells. Cancer Sci 110(2):582–595. https://doi.org/10.1111/cas.13900
Min HK, Kapoor A, Fuchs M, Mirshahi F, Zhou H, Maher J, Kellum J, Warnick R, Contos MJ, Sanyal AJ (2012) Increased hepatic synthesis and dysregulation of cholesterol metabolism is associated with the severity of nonalcoholic fatty liver disease. Cell Metab 15(5):665–674. https://doi.org/10.1016/j.cmet.2012.04.004
Misra HP, Fridovich I (1972) The role of superoxide anion in the autoxidation of epinephrine and a simple assay for superoxide dismutase. J Biol Chem 247(10):3170–3175
Moed L, Shwayder TA, Chang MW (2001) Cantharidin revisited: a blistering defense of an ancient medicine. Arch Dermatol. https://doi.org/10.1001/archderm.137.10.1357
Nájera-Martínez M, Landon-Hernández GG, Romero-López JP, Domínguez-López ML, Vega-López A (2022) (2022) Disruption of neurotransmission, membrane potential, and mitochondrial calcium in the brain and spinal cord of Nile tilapia elicited by Microcystis aeruginosa extract: an uncommon consequence of the eutrophication process. Water Air Soil Pollut. 233(1):6. https://doi.org/10.1007/s11270-021-05480-x
Neumann J, Herzig S, Boknik P, Apel M, Kaspareit G, Schmitz W, Scholz H, Tepel M, Zimmermann N (1995) On the cardiac contractile, biochemical and electrophysiological effects of cantharidin, a phosphatase inhibitor. J Pharmacol Exp Ther 274(1):530–539
Pachuta-Stec A, Szuster-Ciesielska A (2015) New Norcantharidin analogs: synthesis and anticancer activity. Arch Pharm (Weinheim) 348(12):897–907. https://doi.org/10.1002/ardp.201500255
Pan MS, Cao J, Yue-Zu F (2020) Insight into norcantharidin, a small molecule synthetic compound with potential multi-target anticancer activities. Chin Med 15:55. https://doi.org/10.1186/s13020-020-00338-6
Pizzino G, Irrera N, Cucinotta M, Pallio G, Mannino F, Arcoraci V, Squadrito F, Altavilla D, Bitto A (2017) Oxidative stress: harms and benefits for human health. Oxid Med Cell Longev 2017:8416763. https://doi.org/10.1155/2017/8416763
R Core Team (2022) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna. https://www.R-project.org/. Accessed 4 Jul 2022
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nat Methods 9(7):671–675. https://doi.org/10.1038/nmeth.2089
Tinklenberg RL, Murphy SD, Mochel JP, Seo YJ, Mahaffey AL, Yan Y, Ward JL (2020) Evaluation of dose-response effects of short-term oral prednisone administration on clinicopathologic and hemodynamic variables in healthy dogs. Am J Vet Res 81(4):317–325. https://doi.org/10.2460/ajvr.81.4.317
Vroon DH, Israili Z (1990) Aminotransferases. In: Walker HK, Hall WD, Hurst JW (eds) Clinical methods: the history, physical, and laboratory examinations, 3rd edn. Butterworths, Boston (99:492-493)
Whitfield JB (2001) Gamma glutamyl transferase. Crit Rev Clin Lab Sci 38(4):263–355. https://doi.org/10.1080/20014091084227
Xie C, Schwen LO, Wei W, Schenk A, Zafarnia S, Gremse F, Dahmen U (2016) Quantification of hepatic vascular and parenchymal regeneration in mice. PLoS ONE 11(8):e0160581. https://doi.org/10.1371/journal.pone.0160581
Xie MH, Ge M, Peng JB, Jiang XR, Wang DS, Ji LQ, Ying Y, Wang Z (2019) In-vivo anti-tumor activity of a novel poloxamer-based thermosensitive in situ gel for sustained delivery of norcantharidin. Pharm Dev Technol 24(5):623–629. https://doi.org/10.1080/10837450.2018.1550788
Zhang Y, Klaassen CD (2013) Hormonal REgulation of Cyp4a Isoforms in mouse liver and kidney. Xenobiotica 43(12):1055–1063. https://doi.org/10.3109/00498254.2013.797622
Zhou J, Ren Y, Tan L, Song X, Wang M, Li Y, Cao Z, Guo C (2020) Norcantharidin: research advances in pharmaceutical activities and derivatives in recent years. Biomed Pharmacother 131:110755. https://doi.org/10.1016/j.biopha.2020.110755
Funding
This study was supported by Instituto Politécnico Nacional SIP code 20210197, and 20220561. Author Martínez-Razo G. is a DSc. student who received scholarship from CONACyT and BEIFI-IPN. M.L. Dominguez-López, E. Reyes-Maldonado, José M. de la Rosa, Diego A. Fabila-Bustos and A. Vega-López, are fellows of Estímulos al Desempeño en Investigación and Comisión y Fomento de Actividades Académicas (Instituto Politécnico Nacional) and Sistema Nacional de Investigadores (SNI, CONACyT, México).
Author information
Authors and Affiliations
Contributions
GMR conceived, conducted experiments and wrote the manuscript. MLDM, ERM, ECV data curation, formal analysis. JMDLR, DAFB software, validation; visualization. AVL conceived, funding acquisition, project administration, writing—review and editing. All authors read and approved the manuscript and all data were generated in-house and no paper mill was used.
Corresponding author
Ethics declarations
Ethics approval
The study protocol was approved by the local Ethical Committee (license number: PE/001/2574–6/20). Healthy BDF1 mice were obtained from the National School of Biological Sciences, Mexico, and were maintained in agreement with Article 38 and Chapter V of the Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes.
Consent to participate
Not applicable.
Consent for publication
Not applicable.
Competing interests
The authors declare no competing interests.
Additional information
Publisher's note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Martínez-Razo, G., Domínguez-López, M.L., de la Rosa, J.M. et al. Norcantharidin toxicity profile: an in vivo murine study. Naunyn-Schmiedeberg's Arch Pharmacol 396, 99–108 (2023). https://doi.org/10.1007/s00210-022-02299-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00210-022-02299-z