- Theoretical results:

Eight extracts from eight selected medicinal plants were identified to exhibit anti-inflammatory activity through inhibition of nitric oxide (NO) production and suppression of the pro-inflammatory cytokine TNF-α expression. The evaluation results indicated that the leaf extract of Quýt hoi (MC04), the root extract of Nhó đông (MC03), the root extract of Xáo tam phân (MC06), and the root extract of Chanh rừng (MC25) are promising candidates for further in-depth studies on chemical composition and anti-inflammatory activity. For the first time, the leaf extract of Citrus × aurantium was found to strongly inhibit TNF-α and to suppress IL-6 production. In addition, the botanical identity of Quýt hoi was established for the first time using both morphological characteristics and DNA sequencing, confirming its scientific name as Citrus × aurantium L.

For the first time, we demonstrated that the 95% ethanol extract of the roots of Morinda longissima is non-toxic and exhibits strong anti-inflammatory activity in mice.

A total of 13 compounds were isolated, including three compounds—morindone, ostruthin and murrayafoline A—and ten purified compounds from the leaves of Citrus × aurantium identified as tangeretin (1), 5-demethylnobiletin (2), nobiletin (3), hesperetin (4), hesperidin (5), 5-hydroxynoracronycine (6), citracridone I (7), sinensetin (8), naringenin (9), and quercetin (10). Notably, tangeretin (1) exhibited strong inhibitory effects on TNF-α production at 100 µM and 20 µM, while nobiletin (3) inhibited TNF-α at 100 µM.

For the first time, the anti-inflammatory activities of morindone, ostruthin and murrayafoline A were demonstrated in a co-stimulation macrophage model activated by both bacterial and viral agents. These compounds significantly inhibited the expression of pro-inflammatory cytokines (TNF-α, IL-6, IL-10) and downregulated several key inflammatory-related genes, including TLR3, MyD88, NF-κB, AHR, β-actin, Cyp1b1, ARID5A, Jnk, and Stat1.

A scalable extraction process for producing 95% ethanol extract from Citrus × aurantium leaves at a batch size of 15 kg was successfully developed. A draft internal standard (TCCS) for the leaf extract was established and validated. The extract was confirmed to be non-toxic and to exhibit significant anti-inflammatory activity.

The HL capsule formulation was successfully developed from Citrus × aurantium leaf extract, Morinda longissima root extract, curcumin, menthol, and suitable excipients. The HL capsules met all quality parameters according to the Vietnamese Pharmacopoeia V. An internal standard for the HL capsule was also established and validated.

Acute toxicity studies of the HL capsules showed that the median lethal dose (LD₅₀) could not be determined, indicating very low toxicity. The HL capsules caused no sub-chronic toxicity in white rats at doses of 0.388 g/kg/day and 0.766 g/kg/day. At a dose of 400 mg/kg/day (approximately 4 capsules/day in humans), the HL capsules exhibited significant anti-inflammatory effects in an LPS-induced pneumonia model in rats. The capsules reduced pulmonary edema based on the wet/dry lung weight ratio; alleviated lung inflammation and tissue injury as shown by histopathology; and decreased total protein, LDH, and NOx levels in bronchoalveolar lavage fluid.

- Applied results:

A pilot-scale production of the extract-based formulation from Citrus × aurantium leaves (15 kg per batch) was successfully carried out, and the obtained product met all quality requirements specified in the Vietnamese Pharmacopoeia V.

The HL capsule was successfully produced at pilot scale (1,000 capsules per batch), and the finished product complied with all quality standards of the Vietnamese Pharmacopoeia V.

- For the first time, the medicinal plant “Quýt hoi” was taxonomically identified using morphological analysis combined with gene sequencing, confirming that it is a newly derived hybrid belonging to Citrus × aurantium L.

- The extract of Citrus × aurantium L., along with its major constituents tangeretin (QH1) and nobiletin (QH3), was demonstrated to possess strong anti-inflammatory activity.

- For the first time, we demonstrated that the 95% ethanol extract from the roots of Morinda longissima is non-toxic and exhibits potent anti-inflammatory effects in mice.

- For the first time, the anti-inflammatory activities of morindone, ostruthin, and murrayafoline A were confirmed in a macrophage co-stimulation model activated by both bacterial and viral agents.

- For the first time, we successfully developed the HL capsule formulation, in which the principal ingredient—ethanolic leaf extract of Citrus × aurantium—was shown to be non-toxic and effective in reducing LPS-induced pneumonia in mice.

- Published research articles (list):

1. Hafidha Mehallah, Noureddine Djebli; Pham Ngoc Khanh, Nguyen Xuan Ha, Vu Thi Ha, Tran Thu Huong, Do Dinh Tung, Nguyen Manh Cuong; In silico and in vivo study of anti-inflammatory activity of Morinda longissima (Rubiaceae) extract and phytochemicals for treatment of inflammation-mediated diseases, Journal of Ethnopharmacology, 2024, 328, 118051. https://doi.org/10.1016/j.jep.2024.118051. SCI, Q1, IF: 5.4.

2.Trang Thu Tran, Nguyen Manh Cuong, Nguyen Xuan Ha, Taisuke Nakahama, and Nam Trung Nguyen, Murrayafoline A inhibits inflammatory cytokines in RAW264.7 cells stimulated with LPS and poly (I:C) by targeting the aryl hydrocarbon receptor, Bangladesh Journal of Pharmacology, 2025, 20(1), 35-39. SCIE, Q3, IF: 0.9. 

3. Tran Thu Trang, Ta Minh Huyen, Vu Thi Tien, To Minh Nhat, Vu Thanh Loc, Tran Thu Huong, Nguyen Xuan Ha, Hai Pham-The, Nguyen Manh Cuong, Nguyen Trung Nam, Ostruthin targets IL-6 and downregulates Arid5a expression to inhibit cytokine production under TLR3-TLR4 cotriggering: in vitro and in silico studies, Natural Product Communications, 2025, 20(10), 1-10. DOI: 10.1177/1934578X251385033. Q3, SCIE, IF 1.4.

6. Tran Thu Huong, Vu Thanh Loc, Nguyen Xuan Ha, Vu Thi Ha, Pham Ngoc Khanh, Dinh Thi Thu Thuy, Tran Duc Binh, Tran The Bach, Nguyen Thi Phuong Trang, Tran Quoc Toan, Nguyen Manh Cuong, Anti-inflammatory components of the essential oil from the leaves of Citrus × aurantium L. based on network pharmacology study, Vietnam Journal of Chemistry, 2025, 63(992), https://doi.org/10.1002/vjch.70089. Q3.

7. Ta Thi Minh Huyen, Vũ Thị Tiến, Nguyen Trung Nam, Nguyen Manh Cuong, Tran Thu Trang,  Morindone from Morinda longissima inhibits cytokine production in TLR-costimulated macrophage model potentially through JNK/NF-κB/STAT1, TNU Journal of Science and Technology, 2025, 230(13): 144 – 150. https://doi.org/10.34238/tnu-jst.12969. Scopus.

 - Patents (list):

Patent: “Process for isolating the anti-inflammatory compound tangeretin from the stems and leaves of Quýt hoi (Citrus × aurantium L.)”. Application number: 2-2023-00370. Filing date: October 13, 2024. Decision on acceptance of valid application No. 67243/QĐ/SHTT, dated December 15, 2024.

- Technological products (describe in details: technical characteristics, place):

5000 capsults.

- Other products (if applicable): Provided supervision and training for one undergraduate student from Hanoi University of Science and Technology (Nguyen Ngoc Anh Thu, international program)