SP-8356: A Novel Verbenone Derivative Exerts In Vitro Anti-Non-Small Cell Lung Cancer Effects, Promotes Apoptosis via The P53/MDM2 Axis and Inhibits Tumor Formation in Mice

Document Type : Original Article


Department of Clinical Laboratory, People’s Hospital of Chongqing Liang jiang New Area, Chongqing, China


Objective: Non-small cell lung cancer (NSCLC) stands as a prominent contributor to cancer-related fatalities on a
global scale, necessitating the search for novel therapeutic agents. SP-8356, a derivative of (1S)-(–)-verbenone, has
shown promise as an anticancer agent in preclinical studies. However, specific mechanisms underlying its effects in
NSCLC remain to be elucidated. The aim of this research was to explore the in vitro anti-NSCLC effects of SP-8356,
elucidate its mechanisms of action, and assess its efficacy in inhibiting tumor formation in a murine model.
Materials and Methods: In this experimental study, NSCLC cell lines were treated with various concentrations of SP-
8356. Cell viability and proliferation were assessed using MTT and colony formation assays, respectively. Cell cycle
distribution was analyzed by flow cytometry, and apoptosis was evaluated by determining apoptotic protein expression.
Western blot analysis was conducted to assess protein expression levels of the both p53 and MDM2. Additionally, we
evaluated efficacy of the SP-8356 in inhibiting tumor formation of the nude mouse model.
Results: SP-8356 demonstrated a concentration-dependent inhibition of cell proliferation in the NSCLC cell lines. Flow
cytometric analysis showed that SP-8356 led to cell cycle arrest at the G2/M phase, indicating its potential influence
on regulating the cell cycle. SP-8356 treatment was associated with the downregulation of CDK1 and Cyclin B1.
Additionally, SP-8356 significantly enhanced apoptosis in NSCLC cells. SP-8356 treatment was associated with the
downregulation of Bcl-2, while Bax expression was upregulated. Mechanistically, SP-8356 led to accumulation of the
p53 protein levels within the NSCLC cells. This accumulation was mediated through inhibition of its negative regulator,
MDM2. Using a nude mouse model demonstrated that SP-8356 effectively inhibited tumor formation in vivo.
Conclusion: Our findings shed light on the molecular mechanisms underlying anticancer activity of SP-8356 and
highlight its potential as a promising therapeutic candidate for NSCLC treatment.


Main Subjects

  1. Sung H, Ferlay J, Siegel RL, Javersanne M, Soerjomataram I, Jemal A, et al. global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin. 2021; 71(3): 209-249.
  2. Wirsdörfer F, de Leve S, Jendrossek V. Combining radiotherapy and immunotherapy in lung cancer: can we expect limitations due to altered normal tissue toxicity? Int J Mol Sci. 2018; 20(1): 24.
  3. Chen Z, Fillmore CM, Hammerman PS, Kim CF, Wong KK. Non-smallcell lung cancers: a heterogeneous set of diseases. Nat Rev Cancer. 2014; 14(8): 535-546.
  4. Tandon N, Goller K, Wang F, Soibam B, Gagea M, Jain AK, et al. Aberrant expression of embryonic mesendoderm factor MESP1 promotes tumorigenesis. EBioMedicine. 2019; 50: 55-66.
  5. Baslan T, Morris JP 4th, Zhao Z, Reyes J, Ho YJ, Tsanov KM, et al. Ordered and deterministic cancer genome evolution after p53 loss. Nature. 2022; 608(7924): 795-802.
  6. Saleh MM, Scheffler M, Merkelbach-Bruse S, Scheel AH, Ulmer B, Wolf J, et al. Comprehensive analysis of TP53 and KEAP1 mutations and their impact on survival in localized- and advanced-stage NSCLC. J Thorac Oncol. 2022; 17(1): 76-88.
  7. Ni L, Xu J, Zhao F, Dai X, Tao J, Pan J, et al. MiR-221-3p-mediated downregulation of MDM2 reverses the paclitaxel resistance of nonsmall cell lung cancer in vitro and in vivo. Eur J Pharmacol. 2021; 899: 174054.
  8. Atanasov AG, Zotchev SB, Dirsch VM, International Natural Product Sciences Taskforce, Supuran CT. Natural products in drug discovery: advances and opportunities. Nat Rev Drug Discov. 2021; 20(3): 200- 216.
  9. Perricone M, Arace E, Corbo MR, Sinigaglia M, Bevilacqua A. Bioactivity of essential oils: a review on their interaction with food components. Front Microbiol. 2015; 6: 76.
  10. Mander S, Kim DH, Thi Nguyen H, Yong HJ, Pahk K, Kim EY, et al. SP-8356, a (1S)-(-)-verbenone derivative, exerts in vitro and in vivo anti-breast cancer effects by inhibiting NF-κB signaling. Sci Rep. 2019; 9(1): 6595.
  11. Cui W, Yang D, Chen X, Yu H. SP-8356 (a verbenone derivative) inhibits proliferation, suppresses cell migration and invasion and decreases tumor growth of osteosarcoma: role of PGC-1α/TFAM and AMPK-activation. Cell J. 2023; 25(5): 291-299.
  12. Kim DH, Yong HJ, Mander S, Nguyen HT, Nguyen LP, Park HK, et al. SP-8356, a (1S)-(-)-verbenone derivative, inhibits the growth and motility of liver cancer cells by regulating NF-κB and ERK signaling. Biomol Ther (Seoul). 2021; 29(3): 331-341.
  13. Ju C, Song S, Hwang S, Kim C, Kim M, Gu J, et al. Discovery of novel (1S)-(-)-verbenone derivatives with anti-oxidant and antiischemic effects. Bioorg Med Chem Lett. 2013; 23(19): 5421-5425.
  14. Dela Cruz CS, Tanoue LT, Matthay RA. Lung cancer: epidemiology, etiology, and prevention. Clin Chest Med. 2011; 32(4): 605-644.
  15. Bai RL, Chen NF, Li LY, Cui JW. A brand new era of cancer immunotherapy: breakthroughs and challenges. Chin Med J (Engl). 2021; 134(11): 1267-1275.
  16. Pucci C, Martinelli C, Ciofani G. Innovative approaches for cancer treatment: current perspectives and new challenges. Ecancermedicalscience. 2019; 13: 961.
  17. Blagosklonny MV. Selective protection of normal cells from chemotherapy, while killing drug-resistant cancer cells. Oncotarget. 2023; 14: 193-206.
  18. Mansoori B, Mohammadi A, Davudian S, Shirjang S, Baradaran B. The different mechanisms of cancer drug resistance: a brief review. Adv Pharm Bull. 2017; 7(3): 339-348.
  19. Elmore S. Apoptosis: a review of programmed cell death. Toxicol Pathol. 2007; 35(4): 495-516.
  20. Toufektchan E, Toledo F. The guardian of the genome revisited: p53 downregulates genes required for telomere maintenance, DNA repair, and centromere structure. Cancers (Basel). 2018; 10(5): 135.
  21. Marvalim C, Datta A, Lee SC. Role of p53 in breast cancer progression: an insight into p53 targeted therapy. Theranostics. 2023; 13(4): 1421-1442.