{'fraction_non_alphanumeric': 0.04320169626292075, 'fraction_numerical': 0.02941956003180493, 'mean_word_length': 4.970727848101266, 'pattern_counts': {'":': 0, '<': 0, '<?xml version=': 0, '>': 0, 'https://': 0, 'lorem ipsum': 0, 'www.': 0, 'xml': 0}, 'pii_count': 0, 'substrings_counts': 0, 'word_list_counts': {'cursed_substrings.json': 0, 'profanity_word_list.json': 0, 'sexual_word_list.json': 0, 'zh_pornsignals.json': 0}}
Artemisiaside A: A new Lignan Glycoside and its Analogues From the Aerial Parts of Artemisia argyi

Chemical analysis of the aerial parts of Artemisia argyi H. Lév. & Vaniot led to the isolation of 6 lignans, including a new lignan glycoside, artemisiaside A, using various chromatographic techniques. Detailed spectroscopic (including 1D, 2D- nuclear magnetic resonance) and high resolution mass spectroscopy procedures, and electronic circular dichroism were used to ascertain the structural orientations of these compounds. The anti-inflammatory activities of compounds 1 to 6 were evaluated by measuring their inhibitory effects on lipopolysaccharide (LPS) -induced nitric oxide (NO) production in RAW264.7 LPS-activated macrophages. At 50 μM, compound 1 showed moderate anti-inflammatory activity with an inhibition rate of 61.2%.


Introduction
Compositae, the largest family of flowering plants, encompasses more than 1600 genera and 22,000 species, with a worldwide distribution in all continents except Antarctica. 1 This family has enormous economic benefits by providing food, cosmetics, and pharmaceuticals. Artemisia is a large genus in the Compositae family, with more than 350 species distributed across parts of the northern hemisphere temperate regions. 2 Artemisia species have been used to treat a variety of diseases, from simple fevers to malaria. 3,4 Secondary metabolites of Artemisia species reported in the literature include phenolic compounds, alkaloids, terpenoids, sterols, and coumarins. 5 Especially, the discovery of artemisinin from Artemisia annua has brought more attention to this genus. Artemisia argyi, a perennial plant, known as wormwood in traditional Chinese medicines, is mainly found across parts of eastern Asia and Europe. 6 The stems and leaves of A argyi have a particularly strong fragrance. The whole plant was generally used as a Chinese traditional herb to treat warming meridians to dissipate cold, to stop bleeding, as an anti-inflammatory, to relieve coughmelt sputum, and so on. In addition, it can also be used as a dietary source for treating gastric mucosal injury or unhealthy eating habits. 7 Previous study has shown that the leaves and stems of A argyi mainly contain volatile oil, sesquiterpenoids, flavonoids, phenols, alkaloids, and sugars. 8 In our phytochemical investigation of the methanol extract of the aerial parts of A argyi, we report the isolation and structure determination of 3 lignans and 3 lignan glycosides, including 1 previously undescribed compound (1), and 5 known compounds (2)(3)(4)(5)(6). Their anti-inflammatory activities were assessed by measuring the inhibitory effects on lipopolysaccharide (LPS)-induced NO production in macrophages activated by RAW264.7 LPS.

Result and Discussion
Compound 1 was obtained as a colorless amorphous powder. Based on the high resolution electrospray ionization mass spectroscopy (HRESIMS) ion peak at m/z 623.2341 [M-H] − (calcd 623.2340), its molecular formula was determined as C 30 H 40 O 14 , suggesting eleven degrees of unsaturation. Analysis of the 13 C nuclear magnetic resonance (NMR) data (Table 1) with the aid of distortionless enhancement by polarization transfer experiment uncovered the presence of 1 methyl, 4 methylenes (including 3 oxygenated), 11 methines (including 3 aromatics, 5 oxygenated), 10 quaternary carbons (including 9 aromatics, 1 acetyl), and 4 methoxyl groups. The 1 H and 13 C NMR 1 Wuhu Institute of Technology, Wuhu, PR China data ( Table 1) were reminiscent of 2-acetoxymethyl-1,2,3,4-tetrahydro-7-hydroxy-1-(4-hydroxy-3,5 -dimethoxyphenyl)-3-hydroxymethyl-6,8-dimethoxynaphthalene (compound 1b). 9 However, it is noteworthy that there is a slight difference in the carbon chemical shift at C-7 and C-7 ′ , which probably resulted from the introduction of a sugar moiety at C-9. The key 1H detected heteronuclear multiple bond correlation (HMBC) correlations ( Figure 2) of H-9 with C-1 ′′ confirmed that the glycosyl moiety was attached to C-9. The planar structure of 1 was further determined by a comprehensive interpretation of the 1 H-1 H COSY, HSQC, and HMBC spectra. To determine the absolute configuration of 1, it was acid hydrolyzed to obtain an aglycon and sugar. For the aglycone (1c), due to the low amount, the 1 H NMR and 13 C NMR data could not be completed, but CD ( Figure 3) and specific rotation data (Supplementary Figure S22) were provided; its electronic circular dichroism (ECD) spectrum was almost identical to that of urinatetralin, 10 phyltetralin, 10 and strychnovanosides B-C, 11 and the absolute configuration of the sugar was confirmed as D-glucose by comparison with standard glucose using a chiral HPLC column analysis (Supplementary Figure S20). Thus, the absolute configuration of 1 was established as 7 ′ R, 8S, 8 ′ S. Accordingly, compound 1 was established as shown in Figure 1 and was named artemisiaside A.
As a traditional (folk) medicine, A argyi exhibits multifarious bioactivities and is used to control dysmenorrhea, abdominal pain, and inflammation. 15 For instance, the essential oil from A argyi (AAEO) dose-dependently suppressed the release of pro-inflammatory mediators (nitric oxide [NO], PGE2, and ROS) and cytokines (TNF-α, IL-6, IFN-β, and MCP-1) in LPS-induced RAW264.7 macrophages. 16 As well as the crude extract, dehydromatricarin A was also identified as an active component of this plant against inflammation of the lung. 17 Moreover, several sesquiterpenoids obtained from A argyi have shown an enormous potential for inhibitory activity of NO production in vitro. [18][19][20] Therefore, in order to explore and enrich the variety of compounds with anti-inflammatory activity from A argyi, the anti-inflammatory activities of compounds 1 to 6 were evaluated by measuring their inhibitory effects on LPS-induced NO production in RAW264.7 LPS-activated macrophages. At 50 µM, compound 1 displayed moderate activity with an inhibition rate of 61.2% ( Table 2). At the same concentration, the inhibition rate of the positive control (L-NMMA) was 55.5%.

Conclusion
Six lignan analogues, including 1 new lignan glycoside, were obtained from the aerial parts of A argyi. Their structures were determined from extensive spectroscopic and mass

Plant Material
The

Determination of Sugar Components
Compound 1 (2 mg) was refluxed in 10% HCl/dioxane 1:1 (2 mL) for 2 h. The reaction mixture was extracted with ethyl acetate (5 mL × 3) and further purified by silica gel CC using light petroleum-acetone (5:1) to give compound 1c (0.4 mg). The sugar component in the aqueous layer was evaporated under reduced pressure, dissolved in water (1 mL) and subjected to HPLC analysis (Column: ChromCore Suger-10Ca, 6 μm; Mobile Phase: 0.5 mL/min). Standard D-glucose (Sigma) was subjected to the same reaction and HPLC analysis under the same conditions.

Measurement of Nitric Oxide Production
NO production was assayed in supernatants of cultured RAW264.7 cells using an NO assay kit (Beyotime Institute of Biotechnology). Cells were seeded in 96 well culture plates, and pretreated with the tested compounds (50 µM) and stimulated with LPS (1 μg/mL) for 24 h. The supernatant was mixed with an equal volume of Griess reagent (1% sulfanilamide, 0.1% naphthylethylenediamine dihydrochloride, Figure 3. Experimental ECD spectra of compound 1. and 2.5% phosphoric acid) and incubated at room temperature for 24 h. The concentration of nitrite was measured by reading the absorbance at 570 nm. 21,22 L-NMMA was used as a positive control.