Tomatidine Extraction from Various Solvents and Cancer Inhibition Mechanism Through In Silico Analysis
DOI:
https://doi.org/10.48048/tis.2023.5738Keywords:
Alkaloid-steroid, Non-polar solvent, Phytosterol, SHH cascade, TherapeuticAbstract
Tomatidine is an alkaloid-steroid contained in tomato plant organs that potentially developed as an anti-cancer molecule. However, inappropriate use of solvents is frequently becoming an obstacle in the extractions process. In addition, the mechanism of action of tomatidine in the inhibition of cancer cell signaling pathways needs to be investigated. Therefore, this research aims to discover an appropriate solvent to extract tomatidine effectively, and to analyze the inhibition mechanism of tomatidine against the cancer growth pathway. A total of 80 kg of fresh tomatoes were cut into pieces and steamed at 120 °C for 30 min, then mashed and soaked for 3 days in 70 % ethanol, aquadest, petroleum ether, and chloroform separately, continued with GC/MS analysis. Molecular docking was conducted using targeted proteins including B-Raf proto-oncogen serine/ threonine protein kinase (BRAF), Vascular endothelial growth factor A (VEGFA), Smoothened protein (SMO), Cyclin-dependent kinase 2 (CDK2), and Tyrosine-protein kinase/ Janus kinase-1 (JAK1). The ligand-receptor was performed using AutoDock Vina v1.2.0. The highest extracted tomatidine was found in chloroform at 2.44 % of total alkaloid-steroid, and cannot be found in water. The highest interaction is observed in the SMO-tomatidine interaction complex, followed by JAK-1. Furthermore, physicochemical properties of tomatidine make it invaginated into the hydrophobic side of the target proteins that are mainly triggered by van der Waals and alkyl interaction. Based on the in-silico study, tomatidine significantly interacted in the oncogenic signaling pathways that therapeutically potential inhibit cancer cell proliferation. Further study should be focused on the effectiveness analysis of the SHH signaling pathway in the cancer cell.
HIGHLIGHTS
Important findings in this study include:
- Tomatidine, an alkaloid-steroid, is properly extracted in chloroform, petroleum ether and 70 % ethanol, however cannot extracted in aquadest. Furthermore, the aquadest content in 70 % ethanol is likely to reduce the solubility of the tomatidine
- The tomatidine physicochemical property provides a closed covalent bond that makes it more insoluble in the polar solvent. The only polar side is supported by the hydroxyl group in C-4, side chain C-17, and amide side in N-24 that allowed hydrogen bond formation which is relevant to the docking result
- In molecular docking, tomatidine strongly triggers Van der Waal forces, alkyl, and pi interaction than hydrogen bond or ionic force
- Interestingly, ligand-receptor interaction always makes tomatidine invaginated or folded into the protein hydrophobic side and strongly attract non-polar and aromatic amino acid, mostly Val, Ala, Leu, and Phe
- The significant finding is that tomatidine creates valid interactions with CDK2, BRAF, VEGFA, JAK1, and SMO that are involved in cancer cell proliferation, evading apoptosis, and sustained angiogenesis. The highest interaction is observed in the SMO-tomatidine interaction complex, which may affect the Sonic Hedgehog (SHH) signaling pathway (one of the most significant oncogenic pathways in various types of cancer)
- Tomatidine is therapeutically potential to inhibit cancer cell proliferation through protein inhibition activities against the SHH signaling pathway in the cancer cell
GRAPHICAL ABSTRACT
Downloads
References
MDB Elias, FL Oliveira, FCR Guma, RB Martucci, R Borojevic and AJ Teodoro. Lycopene inhibits hepatic stellate cell activation and modulates cellular lipid storage and signaling. Food Funct. 2019; 10, 1974-84.
M Friedman. Anticarcinogenic, cardioprotective, and other health benefits of tomato compounds lycopene, α-tomatine, and tomatidine in pure form and in fresh and processed tomatoes. J. Agr. Food Chem. 2013; 61, 9534-50.
OL Awotedu, UE Okeke, PO Ogunbamowo, OS Ariwoola and TO Omolola. Extraction of phytochemical compounds of Leea guineensis (G. Don) leaves using non-polar and polar solvents, Eur. J. Med. Plants 2020; 31, 24-31.
KM Roopashree and D Naik. Advanced method of secondary metabolite extraction and quality analysis. J. Phcog. Phytochemistry 2019; 8, 1829-42.
C Sukthanarak and C Peerapittayamongkol. The effects of tomatidine in stimulating mitophagy in human fibroblast. In: Proceedings of the 7th International Conference on Biochemistry and Molecular Biology, Bangkok, Thailand. 2021.
P Dey, A Kundu, HJ Chakraborty, B Kar, WS Choi, BM Lee, T Bhakta, AG Atanasov and HS Kim. Therapeutic value of steroidal alkaloids in cancer: Current trends and future perspectives. Int. J. Canc. 2019; 145, 1731-44.
J Fujimaki, N Sayama, S Shiotani, T Suzuki, M Nonaka, Y Uezono, M Oyabu, Y Kamei, H Nukaya, Haruo, K Wakabayashi, A Morita, T Sato and S Miura. The steroidal alkaloid tomatidine and tomatidine-rich tomato leaf extract suppress the human gastric cancer-derived 85As2 cells in vitro and in vivo via modulation of interferon-stimulated genes. Nutrients 2022; 14, 1-14.
S Jeon and MM Kim. Tomatidine inhibits cell invasion through the negative modulation of gelatinase and inactivation of p38 and ERK. Chem. Biol. Interact. 2019; 313, 108826.
MH Hsieh, JS Yang, RC Lin, YH Hsieh, SF Yang, HR Chang and KH Lu. Tomatidine represses invasion and migration of human osteosarcoma U2OS and HOS cells by suppression of presenilin 1 and c-Raf-MEK-ERK pathway. Molecules 2020; 25, 1-13.
NM O’Boyle, M Banck, CA James, C Morley, T Vandermeersch and GR Hutchison. Open babel: An open chemical toolbox. J. Cheminformatics 2011; 3, 1-14.
A Pedretti, A Mazzolari, S Gervasoni, L Fumagalli and G Vistoli. The VEGA suite of programs: An versatile platform for cheminformatics and drug design projects. Bioinformatics 2021; 37, 1174-5.
O Trott and AJ Olson. AutoDock vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 2010; 31, 455-61.
J Eberhardt, D Santos-Martins, AF Tillack and S Forli. AutoDock Vina 1.2.0: New docking methods, expanded force field, and python bindings. J. Chem. Inf. Model 2021; 61, 3891-98.
GM Al-Mazaideh, MH Shalayel, FK Al-Swailmi and SH Aladaileh. Vitamin D is a new promising inhibitor to the main protease (Mpro) of COVID-19 by molecular docking. J. Pharmaceut. Res. Int. 2021; 33, 186-91.
P Kangueane and C Nilofer. Protein-protein and domain-domain interactions. Springer Nature, Singapore, 2018, p. 93-113.
KE Hevener, W Zhao, DM Ball, K Babaoglu, J Qi, SW White and RE Lee. Validation of molecular docking programs for virtual screening against dihydropteroate synthase. Bone 2008; 23, 1-7.
H Nawaz, MA Shad, N Rehman, H Andaleeb and N Ullah. Effect of solvent polarity on extraction yield and antioxidant properties of phytochemicals from bean (Phaseolus vulgaris) seeds, Brazilian J. Pharmaceut. Sci. 2020; 56, e17129.
CF Poole. Influence of solvent effects on retention of small molecules in reversed-phase liquid chromatography. Chromatographia 2019; 82, 49-64.
CM Trivedi and VA Rana. Dielectric properties of amino substituted pyridines in dilute solutions of some non-polar solvents at different temperatures. Indian J. Pure Appl. Phys. 2017; 55, 655-63.
K Bux and ST Moin. Solvation of cholesterol in different solvents: A molecular dynamics simulation study. Phys. Chem. Chem. Phys. 2020; 22, 1154-67.
W Tang and KH Row. Design and evaluation of polarity controlled and recyclable deep eutectic solvent based biphasic system for the polarity driven extraction and separation of compounds. J. Cleaner Prod. 2020; 268, 122306.
RA Moreau, L Nyström, BD Whitaker, JK Winkler-Moser, DJ Baer, SK Gebauer and KB Hicks. Phytosterols and their derivatives: Structural diversity, distribution, metabolism, analysis, and health-promoting uses. Progr. Lipid Res. 2018; 70, 35-61.
B Miras-Moreno, AB Sabater-Jara, MA Pedreno and L Almagro. Bioactivity of phytosterols and their production in plant in vitro cultures. J. Agr. Food Chem. 2016; 64, 7049-58.
OA Rozentsvet, ER Kotlova, ES Bogdanova, VN Nesterov, SV Senik and AL Shavarda. Balance of Δ5-and Δ7-sterols and stanols in halophytes in connection with salinity tolerance. Phytochemistry 2022; 198, 113156.
S Feng, L Wang, P Shao, P Sun and CS Yang. A review on chemical and physical modifications of phytosterols and their influence on bioavailability and safety. Crit. Rev. Food Sci. Nutr. 2021; 62, 5638-57.
MS Uddin, S Ferdosh, MJH Akanda, K Ghafoor, AH Rukshana, ME Ali, BY Kamaruzzaman, MB Fauzi, S Hadijah, S Shaarani and MZI Sarker. Techniques for the extraction of phytosterols and their benefits in human health: A review. Separ. Sci. Tech. 2018; 53, 2206-23.
A Dey and A Mukherjee. Plant-derived alkaloids: A promising window for neuroprotective drug discovery. Elsevier Inc., Netherlands, 2018, p. 237-320.
T Pantsar and A Poso. Binding affinity via docking: Fact and fiction. Molecule 2018; 23, 8.
M Giese and M Albrecht. Alkyl-alkyl interactions in the periphery of supramolecular entities: From the evaluation of weak forces to applications. Chempluschem 2020; 85, 715-24.
CW Fong. Predicting hydrophobic alkyl-alkyl and perfluoroalkyl-perfluoroalkyl van der Waals dispersion interactions and their potential involvement in protein folding and the binding of drugs in hydrophobic pockets. HAL science ouverte, Adelaide, Australia, 2019.
EL Ghisalberti. Steroidal glycoalkaloids: Isolation, structure, analysis, and biosynthesis. Nat. Prod. Comm. 2006; 1, 859-84.
M Arba, Y Yamin, S Ihsan and DH Tjahjono. Computational approach toward targeting the interaction of porphyrin derivatives with Bcl-2. J. Appl. Pharmaceut. Sci. 2018; 8, 60-6.
S Sharma, I Ali and B Bezbaruah. Quantum mechanical study on the effect of pi-pi stacking interactions on phenolic systems and their compatibility with some polar solvents. J. Chem. Chem. Sci. 2017; 7, 1311-30.
N Sawyer, AM Watkins and PS Arora. Protein domain mimics as modulators of protein-protein interactions. Physiol. Behav. 2017; 50, 1313-22.
SN Dishman, CJ Laconsay, JC Fettinger, DJ Tantillo and JT Shaw. Divergent stereochemical outcomes in the insertion of donor/donor carbenes into the C-H bonds of stereogenic centers. ChemRxiv. 2021; 13, 1030-6.
BW Ninham and VA Parsegian. Van der Waals forces: Special characteristics in lipid-water systems and a general method of calculation based on the lifshitz theory. Biophys. J. 1970; 10, 646-63.
S Kawai, AS Foster, T Björkman, S Nowakowska, J Björk, FF Canova, LH Gade, TA Jung and E Meyer. Van der Waals interactions and the limits of isolated atom models at interfaces. Nat. Comm. 2016; 7, 1-8.
RA DiStasio, OAV Lilienfeld and A Tkatchenko. Collective many-body van der Waals interactions in molecular systems. Proc. Natl. Acad. Sci. USA 2012; 109, 14791-5.
SE Song, SK Shin, HW Cho, SS Woo, TK Kwon and DK Song. Tomatidine inhibits tumor necrosis factor-α-induced apoptosis in C2C12 myoblasts via ameliorating endoplasmic reticulum stress. Mol. Cell. Biochem. 2018; 444, 17-25.
SH Choi, JB Ahn, N Kozukue, HJ Kim, Y Nishitani, L Zhang, M Mizuno, CE Levin and M Friedman. Mendel structure-activity relationships of α-, β 1-, γ-, and δ-tomatine and tomatidine against human breast (MDA-MB-231), gastric (KATO-III), and prostate (PC3) cancer cells. J. Agr. Food Chem. 2012; 60, 3891-99.
M Friedman, CE Levin, SU Lee, HJ Kim, IS Lee, JO Byun and N Kozukue. Tomatine-containing green tomato extracts inhibit growth of human breast, colon, liver, and stomach cancer cells. J. Agr. Food Chem. 2009; 57, 5727-33.
SM Ebert, A Al-Zougbi, SC Bodine and CM Adams. Skeletal muscle atrophy: Discovery of mechanisms and potential therapies. Physiology 2019; 34, 232-9.
SP Singh and KV Sashidhara. Lipid lowering agents of natural origin: An account of some promising chemotypes. Eur. J. Med. Chem. 2017; 140, 331-48.
M Diosa-Toro, B Troost, DVD Pol, AM Heberle, S Urcuqui-Inchima, K Thedieck and JM Smit. Tomatidine, a novel antiviral compound towards dengue virus. Antivir. Res. 2019. 161, 90-9.
GB Carballo, JR Honorato, GPFD Lopes and TCLDSE Spohr. A highlight on sonic hedgehog pathway. Cell Comm. Signaling 2018; 16, 1-15.
IF Amalia, A Sayyidah, KA Larasati and SF Budiarti. Molecular docking analysis of α-tomatine and tomatidine to inhibit epidermal growth factor receptor (EGFR) activation in non-small-cell lung cancer (NSCLC). JSMARTech J. Smart Bioprospecting Tech. 2020; 2, 1-6.
T Yu, Q Wu, X You, H Zhou, S Xu, W He, Z Li, B Li, J Xia, H Zhu, Y Zhao, Y Yang and K Chen. Tomatidine alleviates osteoporosis by downregulation of p53. Med. Sci. Mon. Int. Med. J. Exp. Clin. Res. 2020; 26, e9239961 - 9.
X Lin and AA Farooqi. Cucurbitacin mediated regulation of deregulated oncogenic signaling cascades and non-coding RNAs in different cancers: Spotlight on JAK/STAT, Wnt/β-catenin, mTOR, TRAIL-mediated pathways. Semin. Canc. Biol. 2021; 73, 302-9.
KH Lu, Q Wu, X You, H Zhou, S Xu, W He, Z Li, B Li, J Xia, H Zhu, Y Zhao, Y Yang and K Chen. Curcumin analogue l48h37 suppresses human osteosarcoma U2OS and MG-63 cells’ migration and invasion in culture by inhibition of uPA via the JAK/STAT signaling pathway. Molecules 2020; 26, 30.
TK Rimkus, RL Carpenter, S Qasem, M Chan and HW Lo. Targeting the sonic hedgehog signaling pathway: Review of smoothened and GLI inhibitors. Cancers 2016; 8, 22.
KS Jeng, CJ Jeng, WJ Jeng, IS Sheen, SY Li, CM Leu, YG Tsay and CF Chang. Sonic hedgehog signaling pathway as a potential target to inhibit the progression of hepatocellular carcinoma (review). Oncol. Lett. 2019; 18, 4377-84.
A Szczepny, S Rogers, WSN Jayasekara, K Park, RA McCloy, CR Cochrane, V Ganju, WA Cooper, J Sage, CD Peacock, JE Cain, A Burgess and DN Watkins. The role of canonical and non-canonical hedgehog signaling in tumor progression in a mouse model of small cell lung cancer. Oncogene 2017; 36, 5544-50.
AVDS Faria, AI Akyala, K Parikh, LW Brüggemann, CA Spek, W Cao, MJ Bruno, MF Bijlsma, GM Fuhler and MP Peppelenbosch. Smoothened-dependent and -independent pathways in mammalian noncanonical Hedgehog signaling. J. Biol. Chem. 2019; 294, 9787-98.
S Pietrobono, S Gagliardi and B Stecca. Non-canonical hedgehog signaling pathway in cancer: Activation of GLI transcription factors beyond smoothened. Front. Genet. 2019; 10, 556.
M Ruat, L Hoch, H Faure and D Rognan. Targeting of smoothened for therapeutic gain. Trends Pharmacol. Sci. 2014; 35, 237-46.
E Peer, S Tesanovic and F Aberger. Next-generation hedgehog/GLI pathway inhibitors for cancer therapy. Cancers 2019; 11, 538.
Downloads
Published
Issue
Section
License
Copyright (c) 2022 Walailak University
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
