Trilliumosides K and L, two novel steroidal saponins from rhizomes of Trillium govanianum, as potent anti-cancer agents targeting apoptosis in the A-549 cancer cell line

Bashir Ahmad Lone; Misbah Tabassum; Anil Bhushan; Dixhya Rani; Urvashi Dhiman; Ajaz Ahmad; Hilal Ahmad Mir; Prem N Gupta; D M Mondhe; Sumeet Gairola; Prasoon Gupta

doi:10.3389/fchem.2023.1306271

Trilliumosides K and L, two novel steroidal saponins from rhizomes of Trillium govanianum, as potent anti-cancer agents targeting apoptosis in the A-549 cancer cell line

Front Chem. 2023 Dec 22:11:1306271. doi: 10.3389/fchem.2023.1306271. eCollection 2023.

Authors

Bashir Ahmad Lone^#^{1

2}, Misbah Tabassum^#^{1

3}, Anil Bhushan^{1

2}, Dixhya Rani^{1

2}, Urvashi Dhiman^{1

2}, Ajaz Ahmad⁴, Hilal Ahmad Mir⁵, Prem N Gupta^{1

3}, D M Mondhe^{1

3}, Sumeet Gairola^{1

6}, Prasoon Gupta^{1

2}

Affiliations

¹ CSIR-Human Resource Development Centre, Academy of Scientific and Innovative Research, Ghaziabad, India.
² Natural Products and Medicinal Chemistry Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
³ Pharmacology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.
⁴ Department of Clinical Pharmacy, College of Pharmacy, King Saud University, Riyadh, Saudi Arabia.
⁵ Department of Ophthalmology, Pathology, and Cell Biology, Columbia University, New York, NY, United States.
⁶ Plant Science and Agrotechnology Division, CSIR-Indian Institute of Integrative Medicine, Jammu, India.

^# Contributed equally.

Abstract

Two novel steroidal saponins, trilliumosides K (1) and L (2), were isolated from the rhizomes of Trillium govanianum led by bioactivity-guided phytochemical investigation along with seven known compounds: govanoside D (3), protodioscin (4), borassoside E (5), 20-hydroxyecdysone (6), 5,20-hydroxyecdysone (7), govanic acid (8), and diosgenin (9). The structure of novel compounds 1-2 was established using analysis of spectroscopic data including 1D and 2D nuclear magnetic resonance (NMR) and high resolution mass spectrometry (HR-ESI-MS) data. All isolated compounds were evaluated for in vitro cytotoxic activity against a panel of human cancer cell lines. Compound 1 showed significant cytotoxic activity against the A-549 (Lung) and SW-620 (Colon) cancer cell lines with IC50 values of 1.83 and 1.85 µM, respectively whereas the IC50 value of Compound 2 against the A-549 cell line was found to be 1.79 µM. Among the previously known compounds 3, 5, and 9, the cytotoxic IC50 values were found to be in the range of 5-10 µM. Comprehensive anti-cancer investigation revealed that Compound 2 inhibited in vitro migration and colony-forming capability in the A-549 cell line. Additionally, the mechanistic analysis of Compound 2 on the A-549 cell line indicated distinctive alterations in nuclear morphology, increased reactive oxygen species (ROS) production, and decreased levels of mitochondrial membrane potential (MMP). By upregulating the pro-apoptotic protein BAX and downregulating the anti-apoptotic protein BCL-2, the aforementioned actions eventually cause apoptosis, a crucial hallmark in cancer research, which activates Caspase-3. To the best of our knowledge, this study reports the first mechanistic anti-cancer evaluation of the compounds isolated from the rhizomes of T. govanianum with remarkable cytotoxic activity in the desired micromolar range.

Keywords: A-549; BAX; BCL-2; Trillium govanianum; cytotoxicity; saponins; steroidal glycosides.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the Indian Council of Medical Research (ICMR Grant No. 45/23/2022/TRM/BMS), New Delhi, and partial support from SERB, New Delhi (Grant No. EMR/2016/002584) and Researchers supporting project (RSP2023R350), King Saud University, Riyadh, Saudi Arabia.