*Semisynthesis and cytotoxic activities of andrographolide analogues

Figures & data

Figure 1 Andrographolide (1) Note: please refer to appendix

Scheme 1 Synthesis of andrographolide derivatives. Reagents and conditions: (a) ZnCl₂/DMSO/RT/5-6 h (b) pyridinium p-toluenesulphonate/DMSO/60 – 70°C/2-3 h.

Table I.  Dose response parameters of andrographolide and its derivatives in in vitro anticancer pre-screen cell lines¹.

Compounds	GI50		TGI		LC50
	MCF-7	HCT-116	MCF-7	HCT-116	MCF-7	HCT-116
1	6.4 ± 2.1	5.1 ± 0.1	64.7 ± 12.2	15.5 ± 9.3	81.1 ± 17.5	39.5 ± 9.1
2	5.3 ± 0.3	4.8 ± 0.2	9.7 ± 0.3	9.2 ± 0.1	32.3 ± 2.1	9.6 ± 0.1
3	4.2 ± 2.8	3.8 ± 3.5	8.8 ± 1.1	8.1 ± 1.6	9.4 ± 0.3	8.6 ± 1.2
4	5.3 ± 1.6	4.7 ± 0.3	43.2 ± 3.1	9.0 ± 0.5	56.6 ± 4.4	9.6 ± 0.1
5	5.0 ± 1.1	5.2 ± 1.2	27.5 ± 18.0	9.2 ± 0.3	43.7 ± 3.3	9.6 ± 0.1
6	4.9 ± 2.1	3.9 ± 1.2	29.5 ± 15.9	19.2 ± 2.3	42.3 ± 11.2	15.5 ± 6.6
7	6.8 ± 0.4	5.7 ± 1.6	66.6 ± 8.8	28.6 ± 3.3	82.8 ± 4.9	43.8 ± 5.5
8	3.5 ± 1.5	2.3 ± 2.2	9.1 ± 0.3	8.8 ± 2.6	9.5 ± 0.1	10.6 ± 1.8
9	5.2 ± 1.8	3.8 ± 1.7	9.1 ± 1.5	8.7 ± 0.2	9.5 ± 0.3	9.4 ± 0.1
10	5.5 ± 0.4	4.5 ± 2.2	9.4 ± 1.5	9.0 ± 0.9	9.9 ± 3.3	9.5 ± 2.1
11	6.3 ± 1.2	4.5 ± 1.3	61.0 ± 28.0	9.2 ± 0.1	80.1 ± 14.3	9.7 ± 0.1
12	4.1 ± 0.4	4.1 ± 0.4	8.7 ± 0.3	9.4 ± 0.2	9.5 ± 0.1	20.2 ± 18.4
13	7.5 ± 1.0	7.1 ± 0.7	8.8 ± 11.3	66.0 ± 5.6	89.2 ± 6.6	82.9 ± 2.9
14	7.1 ± 1.4	12.4 ± 5.4	69.3 ± 2.5	76.7 ± 6.6	86.8 ± 5.0	87.8 ± 4.1
15	3.6 ± 1.2	4.5 ± 0.9	11.6 ± 5.9	9.1 ± 0.2	25.3 ± 7.4	16.8 ± 8.6
16	18.0 ± 10.1	10.4 ± 4.1	74.9 ± 5.2	75.6 ± 7.3	90.6 ± 5.2	8.5 ± 3.5
17	3.6 ± 0.6	7.7 ± 3.1	8.7 ± 0.4	9.0 ± 0.1	9.5 ± 0.2	9.5 ± 0.1
18	0.7 ± 0.1	2.6 ± 3.3	86.1 ± 6.1	81.5 ± 9.3	96.6 ± 5.7	95.1 ± 8.3
19	4.6 ± 2.8	3.9 ± 1.2	30.5 ± 2.2	12.2 ± 5.3	32.3 ± 10.5	18.2 ± 3.1
20	3.5 ± 0.2	4.9 ± 1.1	8.8 ± 1.1	9.2 ± 0.2	11.1 ± 3.3	9.5 ± 0.1
21	2.3 ± 1.8	5.3 ± 3.8	8.0 ± 0.5	8.5 ± 2.8	10.2 ± 2.2	9.2 ± 0.2
22	4.9 ± 0.4	5.9 ± 0.6	22.4 ± 13.6	37.9 ± 6.3	51.3 ± 16.6	67.5 ± 13.4
23	3.9 ± 0.1	6.6 ± 2.4	8.8 ± 0.8	9.2 ± 0.4	14.1 ± 8.2	19.2 ± 16.8
24	5.1 ± 0.3	6.9 ± 0.7	44.3 ± 4.9	55.6 ± 15.5	72.5 ± 11.2	77.5 ± 7.4
25	4.7 ± 0.6	5.2 ± 0.3	19.9 ± 18.4	14.6 ± 4.7	40.5 ± 4.5	42.7 ± 2.5
26	4.8 ± 1.2	5.8 ± 1.2	79.3 ± 12.2	66.9 ± 6.6	96.8 ± 5.9	87.7 ± 4.9
27	7.1 ± 1.8	5.8 ± 2.6	19.7 ± 6.3	9.2 ± 10.1	12.3 ± 2.1	19.6 ± 9.1
28	7.8 ± 3.1	6.4 ± 1.5	18.8 ± 1.4	8.6 ± 5.2	12.4 ± 9.3	18.8 ± 7.3
29	10.1 ± 1.9	7.0 ± 1.8	41.2 ± 3.9	9.5 ± 10.5	56.6 ± 11.6	9.8 ± 11.1
30	4.7 ± 2.8	5.9 ± 3.5	32.5 ± 10.2	12.2 ± 3.3	37.3 ± 13.5	16.2 ± 7.1
31	2.2 ± 1.3	2.5 ± 1.0	8.0 ± 2.7	8.4 ± 0.2	8.6 ± 0.5	9.1 ± 5.1

¹ Human colon (HCT-116) and breast (MCF-7) cancer cell lines were treated for 72 h with four different concentrations of compounds ranging from 0.1 to 100 μM. MTT [14] assay was used to calculate GI₅₀, TGI and LC₅₀ values (expressed in μM). Values are mean of 3 separate experiments and errors represent the SD values.

Table II.  Inhibition GI₅₀ values of in vitro cancer cell lines by andrographolide derivatives¹.

Panel cell line	1^2	5	6	7	8	10	21	30	31
Leukaemia
CCRF-CEM	3.98	0.281	1.93	2.12	nt	0.333	2.17	0.518	0.244
HL-60(TB)	15.84	9.36	3.05	1.03	0.626	0.918	nt	>100	2.35
K-562	3.16	6.72	3.46	5.87	2.45	4.26	2.14	3.41	2.19
MOLT-4	25.11	14.00	2.95	1.21	0.648	2.17	nt	2.71	1.27
RPMI-8226	5.01	nt	nt	nt	nt	nt	nt	nt	nt
SR	1.00	nt	nt	nt	nt	nt	nt	>100	nt
Non-small cell lung cancer
A549/ATCC	25.11	3.43	2.72	3.77	4.09	3.72	1.40	1.62	2.19
EKVX	31.62	21.50	14.40	19.10	10.60	21.80	11.51	3.13	3.69
HOP-62	31.62	18.10	22.70	25.30	2.35	18.80	1.94	15.20	2.60
HOP-92	25.11	0.172	1.59	0.793	0.674	1.28	15.30	1.31	0.307
NCI-H226	39.81	1.80	1.99	2.30	2.05	2.20	1.51	1.29	1.66
NCI-H23	19.95	16.80	9.58	15.10	1.81	18.90	2.32	3.40	2.18
NCI-H322M	31.62	11.90	11.00	14.80	2.98	14.60	2.46	2.13	2.53
NCI-H460	19.95	19.40	18.60	16.50	3.08	16.80	1.69	4.35	1.99
NCI-H522	10.00	1.86	nt	0.242	2.62	1.29	3.36	nt	0.368
Colon cancer
COLO 205	15.84	1.91	1.43	1.37	1.81	1.76	18.10	0.626	1.56
HCC-2998	25.11	11.00	4.74	4.84	2.00	16.90	2.58	3.11	2.21
HCT-116	15.84	12.70	5.16	21.80	1.91	5.86	1.25	1.97	1.53
HCT-15	15.84	20.40	10.30	14.20	2.39	16.90	2.18	2.34	1.47
HT29	0.501	3.19	2.27	2.55	2.91	4.16	3.02	0.976	1.75
KM12	15.84	2.18	3.58	7.09	2.83	6.84	1.80	1.37	1.92
SW-620	10.00	4.27	3.48	4.21	3.57	4.34	3.71	0.924	2.78
CNS cancer
SF-268	25.11	20.50	20.40	17.30	3.29	16.90	1.76	10.20	1.92
SF-295	19.95	5.58	12.30	12.60	2.49	11.70	12.90	2.28	2.23
SF-539	19.95	2.47	2.19	2.37	2.79	2.92	3.32	2.36	2.24
SNB-19	50.11	4.00	2.72	11.50	2.99	5.08	18.90	2.63	1.91
SNB-75	15.84	1.85	2.11	3.26	2.29	3.45	3.27	1.08	1.15
U251	10.00	18.00	2.30	14.00	1.69	10.70	1.57	1.72	1.46
Melanoma
LOX IMVI	12.58	3.97	2.04	7.81	1.78	2.90	1.51	1.74	1.25
MALME-3M	15.84	nt	nt	nt	nt	nt	nt	nt	nt
M14	15.84	16.00	25.80	27.90	2.60	17.10	1.66	3.07	3.91
SK-MEL-2	15.84	2.31	3.86	2.03	2.77	13.9	4.93	2.23	2.82
SK-MEL-28	12.58	2.04	2.12	2.93	2.89	3.16	2.49	1.92	1.87
SK-MEL-5	15.84	14.30	3.12	15.50	10.50	17.00	7.13	1.82	1.79
UACC-257	19.95	14.20	2.91	11.60	11.70	10.60	1.84	1.93	1.17
UACC-62	19.95	2.78	1.97	5.91	2.50	3.80	nt	1.53	1.77
Ovarian cancer
IGROV1	19.95	2.55	1.31	2.93	2.10	5.33	3.97	0.318	1.85
OVCAR-3	10.00	1.39	2.22	2.46	1.50	2.77	22.40	1.65	1.23
OVCAR-4	10.00	2.94	2.14	9.56	2.50	6.31	nt	1.32	1.71
VCAR-5	15.84	19.00	13.40	16.60	2.80	21.40	2.73	13.70	2.09
OVCAR-8	10.00	3.07	3.48	3.33	2.71	4.61	0.093	0.824	3.03
SK-OV-3	25.11	14.80	15.10	15.80	10.50	13.60	17.60	4.36	3.69
Renal cancer
786-0	5.01	18.10	14.20	19.40	1.92	6.11	1.82	4.17	1.76
A498	31.62	23.90	15.80	25.50	22.00	21.00	23.50	2.97	2.45
ACHN	31.62	18.40	11.00	16.90	1.95	14.80	1.60	2.09	1.65
RXF 393	19.95	Nt	nt	nt	nt	0.013	nt	1.91	nt
CAKI-1	nt	2.43	9.46	10.00	1.62	4.45	19.00	1.25	1.63
SN12C	50.11	3.43	1.68	2.62	1.73	2.37	1.67	0.751	1.68
K-10	15.84	11.80	3.71	19.00	4.39	17.90	2.84	1.75	2.19
UO-31	10.00	1.62	1.93	2.70	2.39	14.60	3.18	1.13	2.65
Prostate cancer
PC-3	19.95	0.631	3.10	4.07	1.62	3.71	7.83	2.22	1.35
DU-145	39.81	10.60	4.14	12.00	3.77	7.52	8.44	1.68	2.05
Breast cancer
MCF-7	15.84	26.30	5.53	13.60	2.89	7.63	0.513	3.38	1.72
NCI/ADR-RES	25.11	7.85	3.14	3.44	1.81	12.9	1.84	3.14	1.92
MDA-MB-231/ATCC	25.11	2.25	2.06	3.00	2.11	2.37	1.91	1.81	1.80
HS 578T	39.81	28.00	7.33	34.60	3.92	20.20	6.85	3.11	2.75
MDA-MB-435	10.00	2.35	4.31	40.13	2.00	3.22	4.15	1.98	1.85
BT-549	7.94	10.20	3.07	9.99	2.90	7.77	3.82	2.16	2.25
T-47D	25.11	1.69	1.30	1.88	1.85	2.26	23.90	0.309	1.14
MG_MID	16.59	5.49	4.26	6.16	2.63	5.49	3.46	2.08	1.77

¹ Data obtained from the NCI's in vitro disease-oriented human tumour cells screen [15]. Values (GI₅₀) are expressed in μM. nt: not tested. Mean panel values (mean graph midpoints MG-MID) of the response parameter were obtained by averaging the individual values for each cell line.

² The values for andrographolide are based on previous screens conducted by the NCI where the highest concentration tested was also 100 μM.

Appendix. Scheme 1. Synthesis of andrographolide derivatives. Reagents and conditions: (a) ZnCl₂/DMSO/RT/5–6 h (b) pyridinium p-toluenesulphonate/DMSO/60–70°C/2–3 h.

Compound No.	Ar1	M.p. (°C)	Yield (%)	Compound No.	Ar1	M.p. (°C)	Yield (%)
2		212–213	62	13		227–228	30
3		228–229	64	14		165–166	91
4		168–169	92	15		172–173	98
5		206–207	67	16		190	77
6		192	32	17		223–224	72
7		179–180	82	18		196–197	74
8		115–116	34	19		185–186	17
9		103–104	89	20		165–166	70
10		206–208	84	21		184–185	81
11		119–120	68	22		169–170	95
12		125–126	99	23		193–194	71
24		143–144	77	28		227–228	85
25		239–240	91	29		226–227	81
26		193–194	65	30		225–228	91
27		219–220	81	31		215–216	78

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