Bioactive glycosides from the roots of Ilex asprella

Figures & data

Table 1. ¹H NMR (pyridine-d₅, 400 MHz) and ¹³C NMR (pyridine-d₅, 100 MHz) data for Compound 1 (δ values in ppm, J values in Hz are in parentheses).

Position	δC	δH	HMBC
1	38.4	α:0.79; β:1.43
2	26.4	α:1.92; β:1.76
3	88.7	3.16, dd(11.2,4.2)
4	39.4
5	55.8	0.75(m)
6	18.6	α:1.43; β:1.29
7	33.1	α:1.94–2.05(m) β:1.43
8	40.1
9	48.2	1.76
10	37.0
11	24.1	1.95
12	123.6	5.50, br s	C-9,14
13	144.2
14	42.0
15	29.0	α:2.36; β:1.27(m)
16	27.9	α:2.84; β:2.11(m)
17	46.4
18	44.5	3.52, br s	C-17
19	80.9	3.58, br s	C-17
20	35.5
21	28.9	2.05
22	33.0	α:2.05; β:1.43
23	28.0	1.21(s)	C-3,4,5,24
24	16.7	0.96(s)	C-3,4,5,23
25	15.4	0.85(s)	C-1, 5, 9,10
26	17.5	1.13(s)	C-7,8,9,11
27	24.8	1.62(s)	C-8,13,14,15
28	177.2
29	28.7	1.15 (s)	C-19,20,21,30
30	24.6	0.98(s)	C-19,20,21,29
3-O-ara	106.4	4.68, d (7.2)	C-3
2′	73.9	4.02
3′	82.4	5.28, t (8.8)	C-4′
4′	74.7	5.18, (td)	C-3′
5′	63.9	α:4.80(dd); β:3.76,(dd)
28-O-glc	95.8	6.37, d (7.9)	C-28
2″	74.1	4.21, t (8.7)
3″	78.8	4.30, t (8.7)
4″	71.0	4.36 (m)
5″	79.2	4.02
6″	62.1	α:4.45(dd); β:4.39,(dd)

Figure 1. Chemical structure of compound 1 isolated from I. asprella.

Figure 2. Chemical structure of compound 2 isolated from I. asprella.

Figure 3. The key HMBC correlation (H–C) of compound 1.

Figure 4. Inhibitory effects of Oseltamivir on the neuraminidase and IP-10 activity. The bar indicates the standard deviation of three different experiments.

Figure 5. Inhibitory effects of compound 1 on the neuraminidase and IP-10 activity. The bar indicates the standard deviation of three different experiments.

Table 2. Cytotoxicity and inhibitory effects of compounds 1 and 2 on neuraminidase activity.

Compound	EC50 (μM)^a	CC50 (μM)^b	SI^c
1	4.1 ± 0.2	>100	>24.3
2	1.7 ± 0.14	>100	>58.8
Oseltamivir^d	0.9 ± 0.07	>100	>111.1

^aThe EC₅₀s were determined using the neuraminidase activity and are presented as the mean ± standard deviation (n = 3).

^bThe CC₅₀s were determined using a MTT assay and are presented as the mean ± standard deviation (n = 3).

^cThe SI represents the ratio of CC₅₀ to EC₅₀ for the A/PR/8/34 strain.

^dThe compound was used as the positive control.

Table 3. Cytotoxicity and inhibitory effects of compounds 1 and 2 on IP-10 activity.

Compound	EC50 (μM)^a	CC50 (μM)^b	SI^c
1	6.6 ± 0.18	>100	>15.5
2	2.5 ± 0.33	>100	>40
Oseltamivir^d	1.1 ± 0.05	>100	>90.9

^aThe EC₅₀s were determined using the IP-10 activity and are presented as the mean ± standard deviation (n = 3).

^bThe CC₅₀s were determined using a MTT assay and are presented as the mean ± standard deviation (n = 3).

^cThe SI represents the ratio of CC₅₀ to EC₅₀ for the A/PR/8/34 strain.

^dThe compound was used as the positive control.

Figure 6. Inhibitory effects of compound 2 on the neuraminidase and IP-10 activity. The bar indicates the standard deviation of three different experiments.

Table 4. Inhibitory effects of compounds 1 and 2 on platelet aggregation.

Compound	Inhibition rate (%)^a
control	0
Ozagrel (50 μM)^b	51± 6
1 (80 μM)	47 ± 8
1 (60 μM)	38 ± 3
2 (80 μM)	0
2 (60 μM)	0

^aThe rates of inhibition were determined using platelet aggregation activity and are presented as the mean ± standard deviation (n = 3).

^bThe compound was used as the positive control.