Anti-inflammatory and antiviral activities of compounds from the fruit of Pouteria caimito

Figures & data

Table 1. ¹³C^a NMR Spectroscopic Data of Compounds 1 − 13 (δ in ppm).

No	1	2	3	4	5	6	7	8	9	10	11	12	13
1	–	38.2	38.6	39.6	47.1	47.1	38.0	37.3	38.2	134.4	132.3	125.2	38.9	Glc 1	105.8
2	75.6	27.0	27.0	28.3	67.1	67.2	26.6	23.6	23.4	138.7	138.0	114.5	26.0	Glc 2	75.9
3	31.6	76.9	76.9	78.4	82.3	82.3	70.2	79.9	79.9	114.9	116.1	145.2	82.5	Glc 3	78.7
4	22.5	38.4	28.4	39.6	38.9	39.1	41.4	37.3	37.5	122.5	130.3	145.8	43.5	Glc 4	71.6
5	112.3	54.9	54.8	56.1	54.8	54.8	47.2	54.5	55.2	121.2	121.4	115.9	47.8	Glc 5	78.3
6	145.9	18.2	17.1	19.1	18.0	18.1	17.5	17.8	17.7	129.9	122.5	121.3	18.4	Glc 6	62.8
7	121.8	32.7	32.8	33.5	32.6	32.9	32.3	32.0	34.8	113.2	121.0	148.8	32.5	Glc 1’	94.8
8	125.9	39.7	41.7	40.0	40.2	39.1	38.8	36.5	40.2	157.2	129.3	113.7	40.1	Glc 2’	75.7
9	146.4	46.8	47.1	48.4	46.8	46.9	47.1	47.0	49.9	111.0	113.3	165.4	48.3	Glc 3’	79.5
10	118.5	36.6	36.4	37.6	37.6	37.7	36.3	36.4	36.6	110.9	141.9	73.1	37.1	Glc 4’	71.1
11	40.2	23.2	22.9	24.0	22.9	23.0	22.1	23.3	21.2	154.4	132.8	37.2	24.0	Glc 5’	78.0
12	21.2	126.7	124.4	122.8	124.5	121.4	121.3	121.4	27.2	57.6	167.2	66.9	122.8	Glc 6’	69.1
13	37.6	138.8	138.4	145.1	138.3	144.1	144.2	144.4	41.5	132.1	151.7	69.4	144.3	Rha 1	101.5
14	33.0	41.1	41.7	42.4	41.7	40.8	41.9	41.3	49.9	122.1	111.5	71.0	42.4	Rha 2	72.2
15	37.6	28.3	27.6	28.6	27.5	27.2	27.3	25.1	30.9	143.0	109.6	35.1	28.7	Rha 3	72.6
16	24.6	25.3	23.9	24.1	23.8	22.7	23.0	23.0	24.3	132.6	157.7	173.1	23.5	Rha 4	73.9
17	37.6	46.9	46.9	46.9	47.0	45.8	45.5	37.7	48.9		56.4	64.0	47.3	Rha 5	69.9
18	32.8	53.3	52.5	42.3	52.4	41.4	40.1	41.9	15.3			30.0	42.1	Rha 6	18.9
19	37.4	71.7	38.3	46.8	38.5	45.5	46.4	46.4	16.4			18.5	46.5	Xyl 1	105.5
20	25.0	41.5	38.4	31.2	38.5	30.4	30.5	30.9	73.0			13.5	30.8	Xyl 2	74.8
21	39.5	26.5	30.3	34.5	30.2	33.4	33.0	33.8	25.2				34.1	Xyl 3	77.6
22	28.1	37.3	36.6	33.5	36.3	32.3	32.1	30.7	41.1				33.1	Xyl 4	71.1
23	22.8	28.2	28.3	29.0	28.8	28.8	64.4	27.8	22.2				64.9	Xyl 5	67.1
24	24.2	16.4	16.1	16.8	17.0	16.9	12.7	17.4	125.2				13.8
25	12.1	15.1	15.3	15.8	17.0	16.3	15.5	21.0	130.0				16.4
26	12.0	16.7	17.0	17.7	17.2	17.1	17.1	16.7	25.5				17.7
27	19.9	23.9	23.3	26.4	23.3	25.7	25.7	25.7	17.5				26.1
28	19.8	178.0	178.6	180.6	178.4	179.5	181.5	67.5	27.7				176.7
29	22.9	26.0	18.0	33.5	16.4	32.2	32.4	33.1	16.0				33.3
30		16.1	21.1	24.0	21.1	23.4	23.5	23.1	16.4				23.5
31								16.5	170.8
32									21.0

^a Measured at 100 MHz.

Figure 1. Chemical structures of compounds 1–13. γ-tocopherol (1), 3β,19α-dihydroxy-urs-12-en-28-oic acid (2), ursolic acid (3), oleanic acid (4), 2α-hydroxy ursolic acid (5), maslinic acid (6), hederagenin (7), erythrodiol-3-acetate (8), 3-O-acetyldammarenediol-II (9), perlolyrine (10), chlorogenic acid butyl ester (11), flazine (12), loniceroside C (13).

Table 3. Inhibitory effects of compounds against LPS-induced NO production in RAW264.7 cells.

Compounds	^aIC50 (μM)	^bCC50 (μM)
Indometacin^c	14.5 ± 2.04	>200
2	34.1 ± 2.58	82.90 ± 3.52
3	34.5 ± 3.74	90.3 ± 4.33
4	>100	>100
5	13.1 ± 0.45	60.4 ± 2.92
6	^dN.A	14.7 ± 0.35
8	>100	>100
9	>100	>100
13	>100	>100

^a Half maximal inhibitory concentration and data were expressed as mean ± SD (n = 3).

^b Concentration cytotoxicity 50% and data were expressed as mean ± SD (n = 3).

^c Positive control.

^d Concentrations below CC₅₀ did not show activity.

Table 4. In vitro anti-RSV and anti-HSV-1 activities of samples.

	RSV		HSV-1
Compounds	^aIC50 (μM)	^bCC50 (μM)	^aIC50 (μM)	^bCC50 (μM)
2	25.0 ± 1.25	>100	12.5 ± 0.63	63.1 ± 0.19
3	25.0 ± 1.56	58.0 ± 0.37	25.0 ± 3.13	47.9 ± 0.51
4	>100	>100	>100	>100
5	12.5 ± 0.23	41.2 ± 0.75	12.5 ± 1.25	26.1 ± 0.16
6	^c N.A	45.8 ± 0.56	^c N.A	24.0 ± 0.79
8	>100	>100	>100	>100
9	25.0 ± 3.20	>100	50.0 ± 2.50	>100
13	50.0 ± 6.25	>100	>100	>100
Ribavirin^d	10.0 ± 2.50	>100	–	–
Acyclovir^d	–	–	1.3 ± 0.31	>100

^a Half maximal inhibitory concentration and data were expressed as mean ± SD (n = 3).

^b Concentration cytotoxicity 50% and data were expressed as mean ± SD (n = 3).

^c Concentrations below CC₅₀ did not show activity.

^d Positive control.

Figure 2. Compounds 2, 3, and 5 significantly inhibited LPS-induced NO production in RAW264.7 cells. (A) RAW264.7 cells were treated with vector (DMSO) or compounds 2, 3, and 5 (3.1, 6.3, 12.5, 25 μM) for 48 h and then cell viability was determined using the Cell Counting Kit-8 (n = 4 replicates per dose). (B-D) RAW264.7 cells were incubated with LPS (2 μg/mLmL) in the presence of compounds 2, 3, and 5. After 48 h, the content of NO in the supernatant was determined by using Griess reagent. (n = 3 per group). Data are mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, and ****p < 0.0001 Two-way (A) or one-way (B to D) analysis of variance (ANOVA) followed by Tukey′s multiple comparison test. ns, not significant.

Table 2. Inhibitory effects of fractions against LPS-induced NO production in RAW264.7 cells.

Extracts	^aIC50 (μg/mL)	^bCC50 (μg/mL)
Petroleum ether fraction	26.0 ± 1.84	>100
Ethyl acetate fraction	>100	>100
n-Butanol fraction	62.9 ± 4.42	>100

^a Half maximal inhibitory concentration and data were expressed as mean ± SD (n = 3).

^b Concentration cytotoxicity 50% and data were expressed as mean ± SD (n = 3).

Table 5. Radical scavenge potential for samples.

Compounds	^aIC50 (μM)
Vitamin E^b	23.9 ± 0.09
2	>400
3	>400
4	>400
5	>400
6	>400
8	>400
9	>400
13	>400

^a Half maximal inhibitory concentration and data were expressed as mean ± SD (n = 3).

^bPositive control.

Table 6. Contents of 2α-hydroxy ursolic acid in P. caimito fruit.

Compound	Equation	Linearity	Limit of quantification (LOQ) (μg/ml)	Limit of detection (LOD) (μg/ml)	Content(mg/kg)
2α-hydroxy ursolic acid	y = 585348x + 366374	0.9998	9.240	0.6160	13.3250 ± 0.1528^a

^a Data was expressed as mean ± SD (n = 6).