Effect of LHS-1269 on morphology and angiogenesis of Lewis lung carcinoma

Background. The new anticancer chemical compound LHS-1269 from the class of indolocarbazoles is undergoing preclinical studies at the N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia. LHS-1269 has a high antitumor activity on transplanted tumors of mice, showed high cytotoxic activity on many tumor cells lines in vitro, and has shown an inhibitory effect on vasculogenic mimicry in tumors in vitro. LHS-1269 does not affect the catalytic activity of human topoisomerases I and IIα. An antiangiogenic mechanism of the drug’s antitumor action is suggested.

Aim. To evaluate the effect of LHS-1269 on the morphological features and angiogenesis of transplanted Lewis lung carcinoma (LLC) in BDF₁ mice.

Materials and methods. In BDF₁ mice (n = 20) with transplanted LLC tumor on days 1 and 3 after 5-fold intraperitoneal administration of LHS-1269 in a single dose of 60 mg (total dose – 300 mg/kg), mice (n = 20) on days 5 and 8 after a single intravenous injection of LHS-1269 at a dose of 100 mg/kg, mice (n = 20) with transplanted LLC tumor without administration of LHS-1269 (control). The assessment of the antitumor effect was carried out according to the criterion of inhibition of tumor growth (%) and the study of the morphological features of the tumors. To assess the effect of LHS-1269 on tumor angiogenesis in LLC tumor sections, a visual calculation of the average number (density) of blood vessels and immunohistochemical detection of expression of the CD31+ endothelial marker were performed.

Results. The LHS-1269 compound in animal groups with 5-fold and 1-fold use caused tumor growth inhibition – 59–70 and 67–79 %, with pronounced morphological changes and tumor cell death. There is an uneven distribution of blood vessels in tumors and surrounding tissues in all groups. LHS-1269 caused a statistically significant decrease in the average number of blood vessels in the tumor both after 5-fold and after 1-fold administration. The statistically significant decrease in the average number of blood vessels in the surrounding connective tissue tumor was observed only after 5-fold administration of the drug. The decrease in the average number of CD31+ endothelial cells after five times intraperitoneal administration was statistically insignificant compared to control (83.1 ± 8.7 and 59.6 ± 18.9, respectively). The increase in this indicator after a single intravenous injection of LHS-1269 was statistically significant.

Conclusion. LHS-1269, when administered five times and once in mice, caused pronounced morphological changes in the form of damage and death of LLC tumor cells. The results of a study of angiogenesis in a tumor do not allow an unambiguous conclusion about the inhibitory effect of LHS-1269 on angiogenesis in LLC tumors in mice.

1. Golubeva I.S., Yavorskaya N.P., Eremina V.I. et al. Antitumor activity of indolocarbazole glycosides. Rossiyskiy bioterapevticheskiy zhurnal = Russian Journal of Biotherapy 2016;15(1):23–4 (In Russ.).

2. Yavorskaya N.P., Golubeva I.S., Ektova L.V. et al. Antitumor activity of indolocarbazole LHS-1269. Rossiyskiy bioterapevticheskiy zhurnal = Russian Journal of Biotherapy 2016;15(1):125–6. (In Russ.).

3. Golubeva I.S., Eremina V.I., Tikhonova N.I. et al. Study of the antitumor activity of indolocarbazole LHS-1269 on lung cancer LLC, modes and methods of administration. Rossiyskiy bioterapevticheskiy zhurnal = Russian Journal of Biotherapy 2018;17(S):21. (In Russ.).

4. Vartanian А., Golubewa I., Shprakh Z. Is vasculogenic mimicry a hallmark of an aggressive tumor? In: Horizons in cancer research. V. 65. Ed. by Y.S. Watanabe. New York: Nova Science Publishers, Inc., 2017. P. 13–33.

5. Zenkov R.G., Fetisov T.I., Ektova L.V. et al. Molecular mechanisms of antitumor activity of derivatives of N-glycosides of indocarbazoles LHS-1208 and LHS-1269. Uspekhi molekularnoy onkologii = Advances in molecular oncology 2018;5(4S):87.

6. Acero N., Braña M.F., Añorbe L. et al. Synthesis and biological evaluation of novel indolocarbazoles with anti-angiogenic activity. Eur J Med Chem 2012;48(2):108–13. DOI: 10.1016/j.ejmech. 2011.11.040

7. Kerbel R.S. Tumor angiogenesis. N Engl J Med 2008;358(19):2039–49. DOI: 10.1056/NEJMra0706596

8. Senchukova M.A., Makarova E.V., Kalinin E.A., Tkachev V.V. Modern ideas about the origin, features of morphology prognostic and predictive significance of tumor vessels. Rossiyskiy bioterapevticheskiy zhurnal = Russian Journal of Biotherapy 2019;18(1):6–15. (In Russ.). DOI: 10.17650/1726-9784-2019-18-1-6-15

9. Ivanov A.I., Puchin’yan D.M., Norkin I.A. The role of endo-thelial cells in angiogenesis. Uspekhi sovremennoy biologii = Advances in Modern Biology 2015;136(5):491–505. (In Russ.).

10. Grigorieva I.N., Kharatishvili T.K., Baryshnikov A.Yu. Vasculogenic mimicry – an alternative mechanism of tumor blood supply? Rossiyskiy bioterapevticheskiy zhurnal = Russian Journal of Biotherapy 2011;10(3):25–9.

11. Guide to chemotherapy of tumor diseases. Ed. by N.I. Perevodchikova. 4rd edn., updated and revised. Moscow: Prakticheskaya Meditsina, 2015. P. 41–48. (In Russ.).

12. Blokhin D.Y., Chmutin E.F., Ivanov P.K. Molecular targets for anticancer therapy: growth factors, angiogenesis and apoptosis. Rossiyskiy bioterapevticheskiy zhurnal = Russian Journal of Biotherapy 2011;10(3):17–24. (In Russ.).

13. Livanova A.A., Deev R.V., Rizvanov A.A. Modern methods for the study of angiogenesis in the experiment. Geni i kletki = Genes and Cells 2015;10(1):115–27. (In Russ.).

14. Gorban N.E., Koryakin O.B., Popov A.M. et al. The role of morphological parameters of intratumoral microvessels and proliferative activity of tumor cells in renal cell carcinoma. Onkourologiya = Oncourology 2012;8(4):10–4. (In Russ.). DOI: 10.17650/1726-9776-2012-8-4-10-14

15. Sprindzukn M.V. Angiogenesis and digital pathology. Available at: https://oncology.ru/specialist/journal_oncology/archive/0211/001/?ysclid=lasqywgmsy404775687. (In Russ.).

16. Nikitenko N.V., Senchukova M.A., Tomchuk O.N. et al. Morphological characteristic and clinical value of different types of tumor vessels in breast cancer. Rossiyskiy bioterapevticheskiy Zhurnal = Russian Journal of Biotherapy 2016;15(3):15–22. (In Russ.). DOI: 10.17650/1726-9784-2016-15-3-22

17. Khochenkov D.A., Saprykina N.S., Baryshnikova M.A. et al. A new mechanism of action of aranose – blockade of neoangiogenesis. Vestnik RONTS im. N.N. Blokhina = Journal of N.N. Blokhin Russian Oncology Center 2015;3:71–5. (In Russ.).

18. Fedoseeva V.V., Postovalova E.A., Khalansky V.A. et al. Druginduced pathomorphosis of glioblastoma 101.8 in Wistar rats treated with doxorubicin bound to poly(lactide-co-glycolide) nanoparticles. Sovremenniye tehnologii v meditsine = Modern Technologies in Medicine 2018;10(4):105–12. (In Russ.). DOI: 10.17691/stm2018.10.4.12

19. Plotnikova N.A., Pyataev N.A., Kanaev P.M. et al. Features of the morphology of Lewis lung carcinoma against the background of correction of tumor growth with melatonin and 3-hydroxypyridine. Fundamentalnie issledovaniya = Basic Research 2014;(10-3):549–52.

20. Dzhadranov E.S., Ergazina M.Zh., Dzhangeldina Z.N., Krasnoshtanov A.V. Influence of cyclophosphane on the development of an experimental transplantable tumor of sarcoma rats 45. Vestnik KazNMU = Bulletin of Kazakh National Medical University 2015;(1):351–3. (In Russ.).

21. Graboviy O.M., Tarasova T.O., Kushubarova M.V. Histological assessment of tumor response to chemotherapy/radiation therapy. Klinichna onkologiya = Clinical Oncology 2012;6(2):138–43. (In Russ.).

22. Galakhin K.A., Yugrinov O.G., Kurik E.G. et al. Chemotherapeutic pathomorphosis of malignant tumors. Ukrainskiy khimioterapevticheskiy zhurnal = Ukrainian Journal of Chemotherapy 2000;4:8–12. (In Russ.).

23. Treshcalina Е.M., Zhukova O.S., Gerasimova G.K. et al. Guidelines for preclinical study of the antitumor activity of drugs. In: Guidelines for conducting preclinical studies of drugs. Part 1. Ed. by A.N. Mironov, N.D. Bunyatyan, A.N. Vasil‘yev et al. Мoscow: Grief i K, 2012. P. 642–57. (In Russ.).

24. Mayo J.G. Biological characterizations of the subcutaneously implanted Lewis lung tumor. Cancer Chemother Rep (2) 1972;3(1):325–30.