In vivo anti-inflammatory activity of caffeoylquinic acid derivatives from Solidago virgaurea in rats

Figures & data

Figure 1. Scheme showing the isolation of compounds 1–4 from the ethyl acetate/n-butanol fraction (EtFr).

Figure 2. HPLC chromatographic profile of the bioactive ethyl acetate/n-butanol fraction (EtFr), showing major compounds (1–4) at Rt 10.2, 10.6. 16.2 and 17.0 min, respectively.

Figure 3. The four compounds isolated from S. virgaurea; 3,5-di-O-caffeoylquinic acid (1), 3,4-di-O-caffeoylquinic acid (2), 3,4,5-tri-O-caffeoylquinic acid (3) and 4,5-di-O-caffeoylquinic acid (4).

Table 1. ¹H NMR chemical shifts (δ in ppm) for compounds (1–4) (DMSO, 300 MHZ, J in Hz).

	Compound
Position	1	2	3	4
2	2.22 (2H, m)	2.11 (2H, m)	2.20 (2H, m)	2.22 (2H, m)
3	5.47 (1H, m)	5.35 (1H, m)	5.42 (1H, m)	4.13 (1H, m)
4	4.01 (1H, dd, 8.1, 3.1)	4.94 (1H, dd, 8.1, 3.1)	4.97 (1H, dd, 8.1, 3.1)	4.95 (1H, dd, 8.1, 3.1)
5	5.44 (1H, m)	4.10 (1H, m)	5.20 (1H, m)	5.27 (1H, m)
6	2.12 (2H, m)	1.99 (m, 6 ax) and 2.03 (m, 6 eq)	2.02 (2H, m)	1.87 (m, 6 ax) and 1.91 (m, 6 eq)
2′	7.07 (1H, d, 2.0)	7.73 (1H, d, 2.0)	7.04 (1H, d, 2.0)	7.73 (1H, d, 2.0)
5′	6.79 (1H, d, 8.1)	6.73 (1H, d, 8.1)	6.78 (1H, d, 8.1)	6.74 (1H, d, 8.1)
6′	6.96 (dd, 8.1, 2.0)	7.02 (dd, 8.1, 2.0)	6.94 (dd, 8.1, 2.0)	7.69 (dd, 8.1, 2.0)
7′	7.62 (d, 15.9)	7.70 (d, 15.9)	7.46 (d, 15.9)	7.49 (d, 15.9)
8′	6.34 (d, 15.9)	6.22 (d, 15.9)	6.24 (d, 15.9)	6.24 (d, 15.9)
2″	7.07 (1H, d, 2.0)	7.73 (1H, d, 2.0)	7.04 (1H, d, 2.0)	7.73 (1H, d, 2.0)
5″	6.79 (1H, d, 8.1)	6.73 (1H, d, 8.1)	6.78 (1H, d, 8.1)	6.74 (1H, d, 8.1)
6″	6.96 (dd, 8.1, 2.0)	7.02 (dd, 8.1, 2.0)	6.94 (dd, 8.1, 2.0)	6.96 (dd, 8.1, 2.0)
7″	7.60 (d, 15.9)	7.66 (d, 15.9)	7.45 (d, 15.9)	7.40 (d, 15.9)
8″	6.30 (d, 15.9)	6.13 (d, 15.9)	6.16 (d, 15.9)	6.10 (d, 15.9)
2‴			6.97 (1H, d, 2.0)
5‴			6.73 (1H, d, 8.1)
6‴			6.97 (dd, 8.1, 2.0)
7‴			7.41 (d, 15.9)
8‴			6.11 (d, 15.9)

Table 2. ¹³C NMR chemical shifts (δ in ppm) for compounds (1–4) (DMSO, 75 MHz).

	Compound
Position	1	2	3	4
1	74.7	76.5	76.3	76.0
2	35.9	36.9	36.7	38.4
3	72.5	72.1	72.1	69.3
4	70.8	75.1	75.7	75.7
5	71.9	65.7	71.9	71.9
6	37.7	41.9	41.7	39.5
COOH	177.4	177.9	177.6	176.8
1′	127.8	127.5	127.8	127.6
2′	115.2	115.2	115.1	115.1
3′	146.5	146.7	146.5	146.6
4′	149.3	149.5	149.7	149.6
5′	116.4	116.5	116.5	116.4
6′	122.9	122.0	122.1	123.1
7′	147.0	147.3	147.2	147.5
8′	115.3	114.9	115.1	114.7
9′	168.8	168.5	168.3	168.2
1″	127.7	127.6	127.7	127.7
2″	115.1	115.1	115.2	115.2
3″	146.5	146.7	146.6	146.7
4″	149.4	149.5	149.4	149.6
5″	116.4	116.4	116.5	116.4
6″	123.0	123.2	123.3	123.2
7″	147.1	147.3	147.1	147.7
8″	115.0	114.9	115.0	114.7
9″	168.4	168.6	168.6	168.5
1‴			127.4
2‴			115.3
3‴			146.6
4‴			149.4
5‴			116.3
6‴			123.6
7‴			147.4
8‴			114.8
9‴			168.5

Table 3. The effect of the bioactive fraction and isolated compounds (1–4) on rat paw volume in carrageenan-induced rat paw oedema model.

	1 h		2 h		3 h
Group	Paw Vol. (mL)	% Oedema Inhibition	Paw vol. (mL)	% Oedema inhibition	Paw vol. (mL)	% Oedema Inhibition
Control	0.66 ± 0.07	–	0.48 ± 0.05	–	0.45 ± 0.04	–
Carrageenan	0.72 ± 0.14	–	0.70^a±0.10	–	0.81^a±0.10	–
Indomethacin (10 mg/kg)	0.69 ± 0.04	40.0	0.59 ± 0.08	50.7	0.56^b±0.04	68.5
4 (25 mg/kg)	0.72 ± 0.04	2.8	0.68^a±0.05	8.5	0.718^a,^b±0.05	27.6
4 (50 mg/kg)	0.71 ± 0.04	5.7	0.67^a±0.03	13.8	0.67^a,^b±0.02	38.3
3 (25 mg/kg)	0.71 ± 0.08	8.5	0.65^a±0.04	21.5	0.64^a,^b±0.03	46.6
3 (50 mg/kg)	0.71 ± 0.06	14.3	0.62 ± 0.07	36.1	0.59^a,^b±0.03	60.7
2 (25 mg/kg)	0.72 ± 0.03	2.8	0.68^a±0.03	7.7	0.67^a,^b±0.02	30.1
2 (50 mg/kg)	0.71 ± 0.03	8.5	0.66^a±0.04	19.2	0.66^a,^b±0.02	42.9
1 (25 mg/kg)	0.72 ± 0.04	2.8	0.68^a±0.05	10.7	0.69^a,^b±0.05	34.7
1 (50 mg/kg)	0.72 ± 0.04	8.6	0.65^a±0.04	19.2	0.67^a,^b±0.03	43.9
EtFr (25 mg/kg)	0.71 ± 0.04	4.3	0.67^a±0.03	15.7	0.65^a,^b±0.05	32.7
EtFr (50 mg/kg)	0.71 ± 0.03	9.5	0.66^a±0.05	18.20	0.67^a,^b±0.03	37.8

Notes: Data are presented as mean ± SEM, n = 6. EtFr: ethyl acetate/n-butanol fraction.

^aSignificantly different from the corresponding control group at p < 0.05.

^bSignificantly different from the corresponding carrageenan-treated group at p < 0.05.

Table 4. Effect of oral administration of the bioactive fraction and isolated compounds (1–4) on TNF-α level in carrageenan-induced rat paw oedema model.

Group	TNF-α conc. (pg/mL) (Mean ± SEM)
Control	4.69 ± 0.63
Carrageenan	84.90^a±3.77
Indomethacin (10 mg/kg)	12.60^b±1.30
4 (25 mg/kg)	48.65^a,^b±4.43
4 (50 mg/kg)	31.56^a,^b±4.51
3 (25 mg/kg)	12.81^b±1.25
3 (50 mg/kg)	8.44^b±0.62
2 (25 mg/kg)	66.35^a,^b±1.30
2 (50 mg/kg)	44.69^a,^b±6.76
1 (25 mg/kg)	45.94^a,^b±5.12
1 (50 mg/kg)	30.76^a,^b±3.51
EtFr (25 mg/kg)	43.43^a,^b±2.31
EtFr (50 mg/kg)	28.86^a,^b±1.21

Notes: Data are presented as mean ± SEM, n = 6. EtFr: ethyl acetate/n-butanol fraction.

^a Significantly different from the corresponding control group at p < 0.05.

^b Significantly different from the corresponding carrageenan-treated group at p < 0.05.

Table 5. Effect of oral administration of the bioactive fraction and isolated compounds (1–4) on IL-1β concentration in carrageenan-induced rat paw oedema model.

Treatments	IL-1β conc. (pg/mL) (mean ± SEM)
Control	29.24 ± 2.91
Carrageenan	118.14***±11.12
Indomethacin (10 mg/kg)	52.91###±5.20
4 (25 mg/kg)	67.98###±6.78
4 (50 mg/kg)	56.72###±5.71
3 (25 mg/kg)	20.36###±1.69
3 (50 mg/kg)	5.83###±0.57
2 (25 mg/kg)	97.30 ± 9.54
2 (50 mg/kg)	60.06###±5.98
1 (25 mg/kg)	68.06###±6.66
1 (50 mg/kg)	59.74###±4.76
EtFr (25 mg/kg)	63.42###±3.29
EtFr (50 mg/kg)	45.58###±5.45

Notes: Data are presented as mean ± SEM, n = 6. EtFr: ethyl acetate/n-butanol fraction.

***p < 0.001 compared to control group.

###p < 0.001 compared to carrageenan-treated group.