- Study on manufacture of nano complex contains malloapelta B-curcumin
- Evaluation of acute and semichronic toxicity of nano complex contain malloapelta B-curcumin.
- Manufacture of product based nano complex contains malloapelta B-curcumin to support the treatment of anticancer.

1. Established a procedure for isolating malloapelta B at a scale of 20 kg of dried leaves, 1.1 kg of malloapelta B has been isolated from 300 kg of dried leaves (content > 95% determined by HPLC).

2. Two pairs of enantiomers (4 compounds) and 16 other compounds have been isolated. 12 new compounds have been identified as malloapeltas C-F (MA1-MA6), malloapeltas I and II (MA7 and MA8), malloapeltas J-L (MA9-MA11), apigenin-6-C-β-D-deoxygalactopyranosyl-8-β-D-xylopyranoside (MA14). The known compounds have been identified as schaftoside (MA15), apigenin-7-O-β-D-glucoside (MA16), apigenin 7-O-β-D-apiofuranosyl (1→2)-β-D-glucopyranoside (MA17), blumenol C glucoside (MA18), and acantrifoside E (MA19).

3. Manufactured of nano complexes contain malloapelta B-curcumin with molar ratios 1:10, 1:50, 1:100, 1:500, and 1:1000. This nano micro-emulsion complex has hydrophilic–lipophilic balance index (HLB) ranging between 13 and 18 is hydrophilic micro-emulsion, non-adhesive. Malloapelta B-curcumin particles is stable in the range from 30 to 35 nm, so it can easily penetrate through the cell membrane to promote efficiency and increase the dispersion in water, thereby increasing the bioavailability.

4. The nano micro-emulsion complex were evaluated cytotoxic activity on for human cancer cell lines, HT29, SW480, AGS, and MKN7. As the results, this nano micro-emulsion complex exhibited cytotoxic activity with IC50 ranging 31.3~76.0 μg/ml. The results indicated MC1C complex (MCP) were chosen for next study.

5. The isolated compounds, MA1-MA8 and MA12 were found to exhibit cytotoxic activity. Electrophysiological study revealed that MA8 is a potent selective ANO1 inhibitor, with an IC50 value of 2.64 μM. Compounds MA2, MA3, and MA12 significantly inhibited ovarian cancer cell line (TOV-21G) via apoptosis induced and NF-κB signaling pathway.

6. The preliminary mechanism study on the inhibiting the growth of cancer cells for the MCP showed that: MCP induced condensation, DNA fragmentation and apoptosis with 45.30% at a concentration of 100 µg/ml and 25.64% at a concentration of 20 µg/mL. MCP at the concentration of 100 µg/ml induced apoptosis with 15.71%, primarily go to late apoptosis, and induced necrosis with 60.10% of cells; At the concentration of 20 µg/ml and 4 µg/ml, MCP induced approximately 15.44% and 10.43% of cells go to apoptosis. MCP at the concentration of 100 µg/ml induced with increase of caspase 3 3.04 times, compared to the control. MCP acted and demonstrated an enhancement of cell stopping capacity in the G2/M phase.

7. The acute toxicity of MCP was studied in rat. No deaths were observed with oral administration of 2000 ~ 20000 mg/kgP. Therefore, the LD50 value was higher than 20,0 g/kgP. The safety dose was found as 4000 mg/kgP for oral administration of rat. The pharmacological dose was 400 mg/kgP.

8. The semichronic toxicity studies at the dose of 100 and 200 mg/kgP/day for 28 days, MCP induced slightly changing of weight (decrease), hematological parameters (decrease), liver enzyme (increase), kidney (decrease) compared to control. However, this change dose not have statistical significance (P>0,05).

9. Studies on anti-ulcer effects of MCP on BALB/c rat showed that MCP at the dose of 200 mg/kg/day inhibited 20.50 % ulcer volume at the day of 28th (P>0,05); decreased leukemia and liver AST indexes as positive direction (P<0.05); did not affect hematological parameters, liver, kidney, and other basic parameters (P>0.05). MCP at the dose of 400 mg/kg/day significantly inhibited 31.21 % ulcer volume at the day of 28th (P<0,05); decreased leukemia and liver AST indexes as positive direction (P<0.05); did not affect hematological parameters, liver, kidney, and other basic parameters (P>0.05).

10. Manufactured the functional food based on nano complex contains malloapelta B-curcumin

Training: 01 PhD student, currently working at Institute of Marine Biochemistry

1. 12 new compounds have been identified as malloapeltas C-F (MA1-MA6), malloapeltas I and II (MA7 and MA8), malloapeltas J-L (MA9-MA11), apigenin-6-C-β-D-deoxygalactopyranosyl-8-β-D-xylopyranoside (MA14).

2. . Manufactured of nano complexes contain malloapelta B-curcumin with molar ratios 1:10, 1:50, 1:100, 1:500, and 1:1000. This nano micro-emulsion complex has hydrophilic–lipophilic balance index (HLB) ranging between 13 and 18 is hydrophilic micro-emulsion, non-adhesive. Malloapelta B-curcumin particles is stable in the range from 30 to 35 nm, so it can easily penetrate through the cell membrane to promote efficiency and increase the dispersion in water, thereby increasing the bioavailability.

3. The nano micro-emulsion complex were evaluated cytotoxic activity on for human cancer cell lines, HT29, SW480, AGS , and MKN7. As the results, this nano micro-emulsion complex exhibited cytotoxic activity with IC50 ranging 31.3~76.0 μg/ml. The results indicated MC1C complex (MCP) were chosen for next study.

4. The isolated compounds, MA1-MA8 and MA12 were found to exhibit cytotoxic activity. Electrophysiological study revealed that MA8 is a potent selective ANO1 inhibitor, with an IC50 value of 2.64 μM. Compounds MA2, MA3, and MA12 significantly inhibited ovarian cancer cell line (TOV-21G) via apoptosis induced and NF-κB signaling pathway.

5. The acute toxicity of MCP was studied in rat. No deaths were observed with oral administration of 2000 ~ 20000 mg/kgP.
6. The semichronic toxicity studies at the dose of 100 and 200 mg/kgP/day for 28 days, MCP induced slightly changing of weight (decrease), hematological parameters (decrease), liver enzyme (increase), kidney (decrease) compared to control. However, this change dose not have statistical significance (P>0,05).

7. Studies on anti-ulcer effects of MCP on BALB/c rat showed that:
MCP at the dose of 200 mg/kg/day inhibited 20.50 % ulcer volume at the day of 28th (P>0,05); decreased leukemia and liver AST indexes as positive direction (P<0.05); did not affect hematological parameters, liver, kidney, and other basic parameters (P>0.05).
MCP at the dose of 400 mg/kg/day significantly inhibited 31.21 % ulcer volume at the day of 28th (P<0,05); decreased leukemia and liver AST indexes as positive direction (P<0.05); did not affect hematological parameters, liver, kidney, and other basic parameters (P>0.05).

8. Manufactured the functional food based on nano complex contains malloapelta B-curcumin

SCIE papers
1) P. V. Kiem, N. X. Nhiem, N. H. Anh, D. T. H. Yen, N. T. Cuong, B. H. Tai, P. H. Yen, N. H. Nam, C. V. Minh, P. T. Chinh, Y. H. Jeon, S. J. Park, S. H. Kim, S. H. Kwon. Enantiomeric chromene derivatives with anticancer effects from Mallotus apelta. Bioorganic Chemistry, 104, 104268 (2020). Q1.
2) Y. Seo, N. H. Anh, Y. Heo, S.-H. Park, P. V. Kiem, Y. Lee, D. T. Yen, S. Jo, D. Jeon, B. H. Tai, N. H. Nam, C. V. Minh, S. H. Kim, N. X. Nhiem, W. Namkung. Novel ANO1 inhibitor from Mallotus apelta extract exerts anticancer activity through downregulation of ANO1. International Journal of Molecular Sciences, 21, 6470 (2020). Q1.

Patent
1) N. X. Nhiem. P. V. Kiem, P. V. Cuong. C. V. Minh, N. H. Nam, T. D. Lam, B. H. Tai, P. H. Yen, D. T. Thao, N. T. Cuong. Manufacture process of nano micro-emulsion complex contains malloapelta B-curcumin. Application number: 1-2020-05880. Date of Application: October 14, 2020. Date of acceptance (November 13, 2020). Decision number 17867w/QĐ-SHTT of Intellectual Property office of Vietnam.
2) N. X. Nhiem. P. V. Kiem, C. V. Minh, P. V. Cuong. N. H. Nam, B. H. Tai, D. T. H. Yen, P. H. Yen, N. H. Anh, P. T. Chinh. Malloapelta and preparative method this compounds from Mallotus apelta. Application number: 1-2020-05879. Date of Application: October 14, 2020. Date of acceptance (December 12, 2020). Decision number 19261w/QĐ-SHTT of Intellectual Property office of Vietnam.
Products: 20,000 capsules contain Malloapelta B-Curcumin nano-emulsion complex.