Project's information

Project's title Synthesis and evaluation of anti-cancer activities of 4-aza-podophyllotoxin derivatives
Project’s code CT0000.03/22-23
Research hosting institution Institute of Chemistry
Project leader’s name Dr. Nguyen Ha Thanh
Project duration 01/01/2022 - 31/12/2023
Project’s budget 1,000 million VND
Classify Grade A
Goal and objectives of the project
- Study on the synthesis of 4-aza-podophyllotoxin derivatives by using multicomponent reactions
- Evaluation of anti-cancer activities of synthesized derivatives
Main results

Synthesized successfully 6 4-aza-podophyllotoxin compounds containing heterocycles in the ring E 5a-f via four-component domino reactions (32-50% yields) and three-component domino reactions (64-83% yields).
- Synthesized successfully 17 4-aza-podophyllotoxin compounds 11a-q in 69-89% yields.
- Synthesized successfully 13 N-arylated-dihydrobenzo[g]quinoline-5,10-diones 16a–m in 69-86% yields. 
- Synthesized successfully 8 phenazine--butyrolactone compounds 8’a-h in 62-80% yields. 
- Synthesized successfully 5 compounds 13’a-e in 62 – 75% yields, starting from phenylglyoxal hydrate derivatives. 
- The cytotoxic activity of synthesized compounds was evaluated against cancer cell lines including epidermoid carcinoma (KB), hepatoma carcinoma (HepG2), non-small lung (LU-1 or A549) and breast (MCF7) cancer cell lines. The results showed that 27/44 compounds exhibited cytotoxicity against tested cancer cell lines, of which 16 compounds showed higher cytotoxic activity than those of the reference ellipticine. Besides that, the toxicicty of compounds on non-cancerous human embryonic kidney (Hek-293) cell line was also evaluated. In general, 6 potent compounds 5c, 11h, 11j, 11k, 16i, and 16m exhibited 10 to 200-fold lower cytotoxicity against Hek-293 cell lines than to some tested cancer cell lines. Although compounds 8’a-h didn’t show good cytotoxicity, they exhibited potential anti-inflammatory activity by inhibiting NO production.
- The initial findings have indicated the anticancer mechanism of 4-aza-podophyllotoxin compounds 11j and 11k by arresting the G2/M phase of the cell cycle, activating caspase-3/7 enzymes, and inducing apoptosis. Additionally, the tubulin inhibition mechanism of these two compounds has also been proposed through molecular docking simulations.

Novelty and actuality and scientific meaningfulness of the results

The project focused on the synthesis of 4-aza-podophyllotoxin derivatives by using multicomponent reactions. The synthesized compounds were evaluated anti-cancer activities by using MTT method (3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazoliumbromid) to assess their cytotoxicity against KB, HepG2, A549 (or LU-1), MCF7 cancer cell lines. The results indicated that several compounds exhibited potential cytotoxicity against tested cancer cell lines. Notably, some compounds displayed higher cytotoxicity than ellipticine  as a reference. Two compounds 11j, and 11k exhibited the most potential activity against LU-1 cells (IC50 < 50 nM) were further studied for their anti-cancer activity using flow cytometry and  molecular docking. The most potent cytotoxic compounds 11j, and 11k were found to possess anti-proliferative activity through concentration-dependently inducing cell cycle arrest at G2/M phase, caspase-3/7 activation, and apoptosis as well. Moreover, molecular docking studies exhibited importance interaction of two compounds against residues in the colchicine-binding-site of tubulin. These preliminary results revealed that podophyllotoxin-naphthoquinone compounds are worthy of further study aiming to develop new potential anticancer agents.
Besides that, the projetct also studied on the synthesis and biological evaluation of phenazine--butyrolactone compounds 8’a-h. Those compounds, which were synthesized by using “one-pot” two-step four-component reactions, exhibited potential anti-inflammatory activity by inhibiting NO production.  Their inhibitory activity was higher than that of L-NG-monomethyl arginine citrate (L-NMMA) as a reference.  Therefore, compounds 8’a-h have potential of further development of anti-inflammatory agents due to their inhibitory effects on NO production. Moreover, several epidemiological studies have demonstrated that inflammation can predispose to tumors, therefore, due to  anti-inflammatory effects of compounds 8’a-h, these compounds may have great potential in cancer prevention and therapy.

Products of the project - New podophyllotoxin derivatives
- The results of the projects was published in 5 scientific papers: 
- Spetral data of 49 synthesized compounds
- Contributed to trainning: 1 doctoral student, 1 master, and 2 bachelors in Chemistry 
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