Isolation of an oleanane-type saponin active from Bellis perennis through antitumor bioassay-guided procedures

Figures & data

Figure 1. Structure of compound 1.

Table 1. Mean number of tumors and % tumor inhibitions of fractions (Fr. A, Fr. B, Fr. C, Fr. D, Fr. E, Fr. F, Fr. G, Fr. H and Fr. I) obtained from the n-butanol fraction of B. perennis flowers at 100, 1000 or 10 000 mg/l and controls (water and camptothecin).

Treatments	Mean number of tumors (±SE)	% Tumor inhibition
Water (negative control)	92 ± 2.1^m	–
Camptothecin (positive control)	0 ± 0^a	100
Fr. A (10 000 mg/L)	56 ± 0.9^i	39.1
Fr. A (1000 mg/L)	60 ± 2.1^ij	34.8
Fr. A (100 mg/L)	62 ± 1.8^ij	32.6
Fr. B (10 000 mg/L)	42 ± 1.3^fg	54.3
Fr. B (1000 mg/L)	56 ± 0.7^i	39.1
Fr. B (100 mg/L)	62 ± 1.9^ij	32.6
Fr. C (10 000 mg/L)	34 ± 1.3^e	63
Fr. C (1000 mg/L)	49 ± 1.7^h	46.7
Fr. C (100 mg/L)	61 ± 1.3^ij	33.7
Fr. D (10 000 mg/L)	44 ± 0.6^fg	52.2
Fr. D (1000 mg/L)	56 ± 1.2^i	39.1
Fr. D (100 mg/L)	58 ± 2.3^i	37.0
Fr. E (10 000 mg/L)	35 ± 2.1^e	62.0
Fr. E (1000 mg/L)	45 ± 0.7^fg	51.0
Fr. E (100 mg/L)	90 ± 0.5^m	2.1
Fr. F (10 000 mg/L)	25 ± 1.2^d	72.8
Fr. F (1000 mg/L)	44 ± 1.6^fg	52.2
Fr. F (100 mg/L)	63 ± 1.3^ij	32.0
Fr. G (10 000 mg/L)	0 ± 0^a	100
Fr. G (1000 mg/L)	21 ± 1.2^c	77.2
Fr. G (100 mg/L)	73 ± 0.7^l	20.7
Fr. H (10 000 mg/L)	2 ± 1.4^ab	97.8
Fr. H (1000 mg/L)	42 ± 2.3^fg	54.3
Fr. H (100 mg/L)	64 ± 1.5^ijk	30.4
Fr. I (10 000 mg/L)	43 ± 1.9^fg	53.2
Fr. I (1000 mg/L)	51 ± 0.8^h	44.5
Fr. I (100 mg/L)	56 ± 0.5^f	56.7

Mean values (± standard error) with the same letter within columns are not significantly different at p > 0.05.

Table 2. Mean number of tumors and % tumor inhibitions of fractions (Fr. G₁, Fr. G₂, Fr. G₃ and Fr. G₄) obtained from Fr. G fraction of B. perennis flowers at 750, 1500 or 3000 mg/l and controls (water and camptothecin).

Treatments	Mean number of tumors (±SE)	% Tumor Inhibition
Water (negative control)	81 ± 1.6^i	–
Camptothecin (positive control)	0 ± 0^a	100
Fr. Gj (750 mg/L)	38 ± 1.4^g	53
Fr. Gj (1500 mg/L)	28 ± 2.3^ef	65
Fr. Gj (3000 mg/L)	7 ± 1.9^b	91
Fr. G2 (750 mg/L)	49 ± 0.7^h	40
Fr. G2 (1500 mg/L)	25 ± 1.9^e	69
Fr. G2 (3000 mg/L)	11 ± 1.8^cd	86
Fr. G3 (750 mg/L)	25 ± 1.3^e	69
Fr. G3 (1500 mg/L)	6 ± 1.7^b	93
Fr. G3 (3000 mg/L)	1 ± 1.5^a	99
Fr. G4 (750 mg/L)	21 ± 2.1^d	74
Fr. G4 (1500 mg/L)	10 ± 0.9^c	88
Fr. G4 (3000 mg/L)	2 ± 0.8^ab	98

Mean values (± standard error) with the same letter within columns are not significantly different at p > 0.05.

Table 3. ¹H, ¹³C-NMR of 1 (¹H: 600 MHz, ¹³C: 150 MHz, MeOH-d₄).

	δH, J (Hz)	δC		δH, J (Hz)	δC
Aglycone			Fuc
1	1.20 dd, 2.8, 13.5	43.7	1	5.43 d, 8.0	93.4
	2.03 m		2	3.81 br.t, 9.7, 8.0	73.4
2	4.21 m	70.4	3	3.89 dd, 9.7, 3.4	80.2
3	3.68 d, 3.5	81.1	4	5.34 d, 3.4	73.3
4	–	42.0	5	3.85 m	69.7
5	1.33 d, 10.6	46.6	6	1.06 d, 6.4	15.1
6	1.41 m	17.6	1-Rha
	1.47 m		1	4.86*	102.7
7	1.41 d, 11.0	32.2	2	3.91 m	70.9
	1.64 dd, 8.4, 11.0		3	3.78 dd, 9.5, 2.7	70.7
8	–	39.5	4	3.38 t, 9.5	72.5
9	1.64 dd, 6.0, 11.0	47.0	5	3.81 m	68.9
10	–	36.3	6	1.24 d, 6.1	16.5
11	1.93 m	23.2	Xyl
	2.00 m		1	4.49 d, 7.6	105.5
12	5.34 d, 2.8	122.1	2	3.36 br.t, 9.0, 7.6	74.9
13	–	143.2	3	3.47 br.t, 9.0, 9.4	82.7
14	–	41.6	4	3.53 br.dt, 9.2, 10.0, 5.3	68.4
15	1.45 dd, 15.0, 2.7	35.0	5ax	3.22 br.t, 11.6, 10.0	65.7
	1.69 dd, 15.0, 3.7		5eq	3.87 dd, 11.6, 5.3	65.7
16	4.46 m	73.2	a-Rha
17	–	48.9	1	5.14 d, 1.1	100.6
18	2.94 dd, 13.8, 3.4	40.9	2	3.90 m	70.3
19	1.06 dd, 12.0, 3.4	46.6	3	3.80 dd, 9.3, 2.8	70.9
	2.30 br.t, 13.8,12.0		4	3.53 br.t, 10.1, 9.3	82.6
20	–	29.9	5	3.77 m	67.8
21	1.17 m	35.1	6	1.32 d, 6.1	17.0
	1.92 m		Ara
22	1.77 m	30.5	1	5.05 d, 1.6	110.2
	1.91 m		2	4.00 dd, 3.8, 1.6	82.2
23	3.27 d, 11.1	64.3	3	3.88 dd, 6.2, 3.8	76.7
	3.33 d, 11.1		4	3.92 m	83.9
24	0.91 s	13.4	5a	3.59 dd, 12.2, 4.1	60.8
25	1.33 s	17.0	5b	3.71 dd, 12.2, 3.1	60.8
26	0.80 s	16.5	b-Rha
27	1.39 s	25.7	1	5.14 d, 1.0	101.1
28	–	175.8	2	3.94 dd, 3.0, 1.0	70.8
29	0.88 s	31.9	3	3.70 dd, 9.3, 3.0	70.7
30	0.95 s	23.6	4	3.39 br.t, 9.9, 9.3	72.5
acetyl			5	4.00 m	68.5
Ac-1	2.14	19.4	6	1.24 d, 6.5	15.1
Ac-2	–	171.1

*Overlapping signals.