Immunomodulatory activities of isolated compounds from the root-bark of Cussonia arborea

Figures & data

Figure 1. Chemical structures of isolated compounds from C. arborea.

Table 1. ¹H and ¹³C NMR spectra data of Compound 1 (400 and 100 MHz, CD₃OD) compared with ¹³C NMR data of literature.

Position	^1H δ (multiplicity)	^13C	^13C*	DEPT
1.	1.61 (m)	39.47	38.1	CH2
2.		25.31	25.3	CH2
3.	3.57 (m)	73.78	73.9	CH
4.	–	42.95	43.3	C
5.		48.2	overlapped^a	CH
6.	1.52 (m)	19.11	19.11	CH2
7.		33.46	33.8	CH2
8.	–	37.90	37.9	C
9.		overlapped^a	overlapped^a	CH
10.	–	37.84	37.8	C
11.		24.36	24.4	CH2
12.	5.21 (m)	123.60	123.6	CH
13.	–	143.25	143.3	C
14.	–	43.29	43.0	C
15.		28.82	27.5	CH2
16.		24.51	23.9	CH2
17.	–	47.62	47.2	C
18.	2.83 (dd)	42.72	42.7	CH
19.		47.22	47.6	CH2
20.	–	34.87	33.5	C
21.		33.82	34.9	CH2
22.		32.00	33.9	CH2
23.	3.50 (d)	67.18	67.3	CH2
24.	0.69 (s)	12.74	12.8	CH3
25.	0.97 (s)	16.25	16.4	CH3
26.	0.81 (s)	17.74	17.8	CH3
27.	1.12 (s)	24.04	24.1	CH3
28.	–	181.87	181.7	C
29.	0.90 (s)	33.56	33.6	CH3
30.	0.93 (s)	21.58	21.6	CH3

* He et al. (2003).

^a overlapped with signals of MeOD.

Table 2. ¹H and ¹³C NMR spectra data of Compound 2 (400 and 100 MHz, ppm in CDCl₃) compared with ¹³C NMR data of literature.

Position	^1H δ (multiplicity)	^13C	^13C*	DEPT
1.	1.57 (m)	38.36	38.5	CH2
2.	1.57 (m)	27.14	27.4	CH2
3.	3.18 (m)	78.99	78.7	CH
4.	–	38.73	38.7	C
5.	0.7 (m)	55.17	55.2	CH
6.	1.52 (m)	18.26	18.3	CH2
7.		32.58	32.6	CH2
8.	–	39.20	39.3	C
9.	1.50 (m)	47.59	47.6	CH
10.	–	37.04	37.0	C
11.	1.57 (m)	22.90	23.1	CH2
12.	5.26 (m)	122.61	122.1	CH
13.	–	143.55	143.4	C
14.	–	41.55	41.6	C
15.		27.64	27.7	CH2
16.	1.85 (m)	23.36	23.4	CH2
17.	–	46.48	46.6	C
18.	2.78 (m)	40.96	41.3	CH
19.	1.57 (m)	45.82	45.8	CH2
20.	–	30.66	30.6	C
21.		33.75	33.8	CH2
22.		32.39	32.3	CH2
23.	0.96 (s)	28.07	28.1	CH3
24.	0.75 (s)	15.54	15.6	CH3
25.	0.88 (s)	15.30	15.3	CH3
26.	0.73 (s)	17.09	16.8	CH3
27.	1.11 (s)	25.92	26.0	CH3
28.	–	183.13	181.0	C
29.	0.88 (s)	33.06	33.1	CH3
30.	0.89 (s)	23.55	23.6	CH3

* Mahato and Kundu (1994).

Table 3. ¹H and ¹³C NMR spectra data of Compound 3 (400 and 100 MHz, CDCl₃) compared with ¹³C NMR data of literature.

Position	^1H δ (multiplicity, J)	^13C	^13C*	DEPT
1.	1.84, 1.05	37.26	37.6	CH2
2.	1.47	31.68	31.9	CH2
3.	3.46 (m)	71.81	72.0	CH
4.	2.21 (m)	42.32	42.5	CH2
5.		140.76	140.8	C
6.	5.32 (t)	121.70	121.8	CH
7.		31.68	32.1	CH2
8.	1.80	31.91	32.2	CH
9.	0.9	50.17	50.5	CH
10.		36.52	36.5	C
11.		21.08	21.2	CH2
12.	1.92, 1.13	39.69	40.0	CH2
13.		42.22	42.2	C
14.	0.99	56.88	57.1	CH
15.		24.36	24.5	CH2
16.	1.67	28.91	28.9	CH2
17.	1.33	55.97	56.3	CH
18.		12.05	12.2	CH3
19.	0.99	19.40	19.5	CH3
20.	1.98	40.48	40.4	CH
21.		21.09	21.4	CH3
22.	5.10 (dd, 15.2, 8.4 Hz)	138.31	138.3	CH
23.	4.96 (dd, 15.2, 8.8 Hz)	129.29	129.7	CH
24.	1.51	51.24	51.5	CH
25.		31.91	32.2	CH
26.	0.82	21.21	21.2	CH3
27.	0.76	18.98	19.2	CH3
28.	1.41, 1.13	25.40	25.4	CH2
29.	0.78	12.24	12.2	CH3

* Forgo and Kövér (2004).

Table 4. ¹H and ¹³C NMR spectra data of Compound 4 spectra (400 and 100 MHz, C₅D₅N).

Position	^1H δ (multiplicity, J)	^13C	DEPT
1.	1.72	37.50	CH2
2.	1.47	32.12	CH2
3.	4.28 (m)	72.97	CH
4.	2.4 (m)	39.36	CH2
5.		140.94	C
6.	5.34 (t)	121.93	CH
7.		32.10	CH2
8.	1.3	32.07	CH
9.	0.88	50.37	CH
10.		36.96	C
11.		22.94	CH2
12.	1.92, 1.13	39.96	CH2
13.		42.37	C
14.	0.99	56.94	CH
15.		24.56	CH2
16.	1.67	29.34	CH2
17.	1.33	56.09	CH
18.	0.64	12.16	CH3
19.	0.93	19.44	CH3
20.	2.00	40.80	CH
21.	1.06	21.49	CH3
22.	5.18 (dd, 15.2, 8.4 Hz)	138.85	CH
23.	5.02 (dd, 15.2, 8.8 Hz)	129.49	CH
24.	1.50	51.44	CH
25.		32.91	CH
26.	0.86	19.20	CH3
27.	0.76	14.26	CH3
28.	1.25	25.72	CH2
29.	0.88	12.54	CH3
1′	5.05 (d, 7.8 Hz)	102.59	CH
2′	3.92	78.10	CH
3′	4.25	78.64	CH
4′	4.04	75.37	CH
5′	4.25	78.52	CH
6′	4.28	71.72	CH2

Table 5. ¹H NMR data of Compound 5 (500 MHz, CDCl₃) compared with ¹H NMR data of literature.

Position	^1H δ (multiplicity, J)	^1H* δ (multiplicity, J)
1.	1.27 (m)
2.	2.00 (m)
3.	–	–
4.	–	–
5.
6.
7.
8.	–	–
9.
10.	–	–
11.	1.90 (m)
12.	5.23 (s)	5.23 (s)
13.	–	–
14.	–	–
15.
16.
17.	–	–
18.	2.15 (d, 11.5 Hz)
19.
20.
21.
22.
23.	3.40 (d, 10.5 Hz) 3.70 (d, 10.5 Hz)	3.40 (d, 11.3 Hz) 3.62 (d, 11.3 Hz)
24.	0.77 (s)	0.82
25.	0.87 (s)	0.98
26.	1.06 (s)	1.10
27.	1.23 (s)	1.12
28.	–	–
29.	0.83 (d, 6.5 Hz)	0.83 (d, 6.4 Hz)
30.	0.92 (d, 6.5 Hz)	0.93 (d, 6.2 Hz)

* Fourie et al. (1989).

Figure 2. The graph represents the effect of compounds 1–4 on oxidative burst. Compounds were tested on three different concentrations (1, 10 and 100 µg/mL). Results are presented in relative light units (RLU) and oxidative burst activity of whole blood using luminol as a probe. Each vertical bar represents a mean of triplicate. Error bars represent standard deviations of the means. Significance difference was calculated using one-way ANOVA and * represent p < 0.05 significance difference was compared to the + ve control. Where + ve = cells + zymosan and − ve = cells alone.

Figure 3. Effect of compounds on T-cell proliferation. Compounds were tested on three different concentrations (1, 10 and 100 µg/mL). Results are presented in counts per minutes (CPM). Each vertical bar represents a mean of triplicate. Error bars represent standard deviations of the means. Significance difference was calculated using one-way ANOVA. Significance difference was compared to the control having cells in the presence of PHA. Where * represent p < 0.05.

Table 6. Effect of pure compounds from C. arborea on phagocytes oxidative burst, T-cell proliferation and cytotoxicity on NIH-3T3 cells. The IC₅₀ (µg / mL) was calculated using three doses (1, 10 and 100 µg/mL) of each compound. Values are expressed as mean ± SD of three determinations.

Compounds	ROS inhibition (IC50± SD µg/mL)	T-cell proliferation inhibition(IC50± SD µg/mL)	Cytotoxicity on NIH 3T3-cells(IC50± SD µg/mL)
1	24.4 ± 4.3	>100	>100
2	37.5 ± 0.1	12.6 ± 0.4	>100
3	>100	86.8 ± 0.1	>100
4	>100	>100	>100
5	NT	NT	NT
Ibuprofen	11.2 ± 1.9	–	–
Prednisolone	–	<0.62	–
Cyclohexamide	–	–	0.13 ± 0.02

NT means not tested.

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