EV71(IC50= 31.83 μg/ml )

Norwogonin (0.4, 2, 10, and 50 μg/mL; 48 hours) inhibits EV71 replication in Vero cells and prevents the cytotoxicity induced by EV71 infection[1].
Norwogonin (50 μg/mL; 48 hours) dramatically decreases viral VP2 protein expression[1].
At 48 hours, norwogonin (50 μg/mL) significantly reduces the expression of the EV71 NCR gene[1].

Results: Haungqing (Scutellariae Radix) and baishao (Paeoniae Radix Alba) herb pair (HBHP) significantly reduced DAI score and decreased colon length shortening in DSS-induced UC mice. The administration of HBHP was able to effectively alleviated mucosal ulceration and epithelial destruction. In addition, HBHP treatment obviously - reduced the expressions of TNF-α, IL-6, and IL-1β in colon tissues (p < 0.05 or p < 0.01). 35 bioactive compounds and 290 HBHP targets related to UC were obtained. Among them 3 key active compounds (baicalein, panicolin, and norwogonin) with higher degree values in the drug-compound-target network and 21 hub genes (STAT3, JAK2, SRC, AKT1, PIK3CA, and VEGFA, etc.) were identified. KEGG enrichment analysis suggested that HBHP's mechanisms mainly involve the JAK-STAT pathway. Abnormal activation of JAK/STAT signaling is believed to be involved in the pathogeneses of UC. Notably, WB and IHC showed that HBHP significantly down-regulated the protein expression levels of p-JAK2 (p < 0.05) and p-STAT3 (p < 0.05 or p < 0.01). JAK2 and STAT3 might be core targets for the action of HBHP; this possibility was also supported by molecular docking.[2]
Conclusions: HBHP could alleviate DSS-induced UC, reduce tissue inflammation, and its mechanism might primarily be achieved by inhibiting JAK2/STAT3 signaling pathway. Meanwhile, our work revealed that network pharmacology combined with experimental verification is a cogent means of studying the mechanism of TCM.[2]

In the present study, the anti-EV71 activity of norwogonin, oroxylin A, and mosloflavone from Scutellaria baicalensis Georgi was evaluated using a cytopathic effect (CPE) reduction method, which demonstrated that all three compounds possessed strong anti-EV71 activity and decreased the formation of visible CPEs. Norwogonin, oroxylin A, and mosloflavone also inhibited virus replication during the initial stage of virus infection, and they inhibited viral VP2 protein expression, thereby inhibiting viral capsid protein synthesis. However, ribavirin has a relatively weaker efficacy compared to the other drugs. Therefore, these findings provide important information that will aid in the utilization of norwogonin, oroxylin A, and mosloflavone for EV71 treatment[1].

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Assays of antiviral activity[1]
Assays of antiviral activity were performed with the SRB method that assesses cytopathic effect (CPE) reduction, which was previously reported by Choi et al. (2009a). The effect of norwogonin, oroxylin A, and mosloflavone on EV71-induced CPE was observed. Briefly, Vero cells were seeded onto a 96-well culture plate at a concentration of 2×104 cells per well. Next day, the medium was removed and washed with PBS, and 0.09 mL of diluted virus suspension of EV71 containing TCID50 (50% tissue culture infective dose) of the virus stock to produce an appropriate cytopathic effects within 2 days after infection and 0.01 mL of norwogonin, oroxylin A, mosloflavone, and ribavirin (0.4, 2, 10, and 50 μg/mL, respectively) were added. After incubation at 37°C in 5% CO2 for 2 days, the morphology of the cells was observed under a microscope at 32×10 magnifications, and the images were recorded.

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Time-of-addition assays[1]
The time-of-addition effects of norwogonin, oroxylin A, and mosloflavone were examined in accordance with previously described procedures (Chiang et al., 2003) with minor modifications. Vero cells were seeded onto 96-well culture plates at a density of 2×104 cells per well and incubated for 1 day. After washing with phosphate-buffered saline (PBS), 50 μg/mL of norwogonin, oroxylin A, and mosloflavone were then added to the cells either before (−1 h), during (0 h), or after (1, 2, 4, 6, and 8 h) EV71 infection. After 2 days, the antiviral activity was tested using the SRB assay, and ribavirin was used as the positive control.

First, The UC model of mice induced by dextran sulfate sodium (DSS) was established. The mice were randomly divided into Control group, DSS group, SASP group (390 mg/kg), and HPHP group (1.95 g/kg), with 8 mice per group. Drugs were administrated via oral gavage for 7 days. Then, Disease activity index (DAI), length of the colon, histopathology, and changes in inflammatory cytokines in colonic tissues were analyzed to assess the effect of HBHP on UC. Besides, Network pharmacology was applied to identify the active compounds, core targets of HBHP in the treatment of UC, and the corresponding signaling pathways to explore the underlying mechanisms. Finally, Western blot (WB), immunohistochemistry (IHC) and molecular docking were performed to validate the results.[2]

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UC was induced in mice by administration of 3.5 % (w/v) DSS (MW 36–50 kDa) in drinking water for 7 days ad libitum as described previously. Mice were randomly divided into four groups (n = 8 each): Control group (normal drinking water + normal water orally once daily), DSS group (DSS drinking water + normal water orally once daily), SASP group (DSS drinking water + 390 mg/kg SASP orally once daily), and HPHP group (DSS drinking water + 1.95 g/kg HBHP orally once daily). The dosage of HBHP used here was calculated in proportion to human clinical doses (15 g/day per adult) using body surface area conversion.[2]
DSS solution was updated every day. The oral gavage administration was initiated simultaneously with DSS drinking water. Body weights, stool properties, and stool occult blood were monitored every day. At the end of the experiment, all mice were sacrificed and colonic specimens were collected. The lengths of the colons were measured and then immediately stored at −80 °C or fixed in 10 % paraformaldehyde for further procedures.[2]

[1]. Inhibitory Effects of Norwogonin, Oroxylin A, and Mosloflavone on Enterovirus 71. Biomol Ther (Seoul). 2016 Sep 1;24(5):552-8.

[2]. Heliyon. 2023 Nov 30;9(12):e23082. doi: 10.1016/j.heliyon.2023.e23082. eCollection 2023 Dec.

Norwogonin is a trihydroxyflavone with the hydroxy groups at positions C-5, -7 and -8. It has a role as an antioxidant and a metabolite.
Norwogonin is a natural product found in Scutellaria discolor, Scutellaria scandens, and other organisms with data available.

C15H10O5

270.2369

270.053

4443-09-8

Typically exists as yellow to orange solids at room temperature

1.548g/cm3

528.4ºC at 760 mmHg

258 ºC

206ºC

1.732

Scutellaria baicalensis Georgi

2.6

90.90

C1=CC=C(C=C1)C2=CC(=O)C3=C(O2)C(=C(C=C3O)O)O

ZFKKRRMUPBBYRS-UHFFFAOYSA-N

InChI=1S/C15H10O5/c16-9-6-11(18)14(19)15-13(9)10(17)7-12(20-15)8-4-2-1-3-5-8/h1-7,16,18-19H

5,7,8-trihydroxy-2-phenylchromen-4-one

5,7,8-Trihydroxyflavone;5,7,8-Trihydroxy-2-phenyl-4H-chromen-4-one;FLAVONE, 5,7,8-TRIHYDROXY-; NSC-128304;

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~250 mg/mL (~925.10 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (7.70 mM) (saturation unknown) in 10% DMSO + 40% PEG300 +5% Tween-80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 + to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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 (Please use freshly prepared in vivo formulations for optimal results.)