Table 8.

Pharmacological activity of isolated C. nutans compounds.

Experiment design  (Experiment model; Assay; Incubation period; Test subject; Extract dose; Positive control)	Isolated compound	Result	Reference
(a) Anti-inflammatory activity
in vitro; Prostaglandin E2 determination assay by the radioimmunoassay Immortalized COX-1 and COX-2 mouse lung fibroblast cell; Aspirin (IC50: 2.06 μg/mL (COX-1), 3.57 μg/mL (COX-2))	Cerebrosides	Ineffective	[55]
(b) Immunomodulating activity
in vitro; ConA-induced T cell, LPS-induced B cell, evaluation of Th1 cytokines (IL-2 and IFN-γ), Th2 cytokines (IL-4 and IL-10); Splenocytes; Control on Th2 cytokines IL-4 (138.3 pg/mL) and IL-10 (1234.9 pg/mL)	(1) CN1 (shaftoside)  (2) CN2 (stigmasterol)  (3) CN3 (β-sitosterol)   (4) CN4 (lupeol)	CN3 inhibit T lymphocyte proliferation the most (RP: 0.16) followed by CN2 (RP: 0.47), only CN1 inhibit B cell proliferation (RP: 0.77), all ineffective on Th1 cytokines, CN3 inhibit secretion of IL-4 (22.6 pg/mL) and IL-10 (63.9 pg/mL), CN3 significantly reduce activated helper T cells (54.3%) and activated cytotoxic T cells (62.2%)	[45]
(c) Anti-oxidant activity
in vitro; DPPH scavenging assay; Vitamin C (IC50: 22.589 μg/mL)	(1) Stigmasterol-β-D-glucoside  (2) 3-amino-4,5-dihydroxyfuran-2(3H)-one	Compound 1: Ineffective   Compound 2: IC50: 102.949 μg/mL	[49]
in vitro; DPPH scavenging assay, FRAP assay; 1000 μg/mL (stock); BHT (DPPH), Ascorbic acid (FRAP)	(1) Clinamide D (2) Clinamide E	(1) 76.05 ± 0.02%, (2) 72.84 ± 0.01% of DPPH inhibition, (1) 15.47 ± 0.03%, (2) 38.56 ± 0.02% of FRAP inhibition	[53]
(d) Anti-viral activity
Anti HSV-1 assay  in vitro; HSV-1 virus strain; Vero cell; Acyclovir (IC50: 2–5 μg/mL)	Cerebrosides	Ineffective	[55]
Anti HSV-1 assay  in vitro; HSV-1F strain; Plaque reduction assay-direct, pre, post; 72 h; Vero cell; Acyclovir, Dextran sulfate (1 mg/mL)	(1) 132-hydroxy-(132-R)-phaeophytin b  (2) 132-hydroxy-(132-S)-phaeophytin a  (3) 132-hydroxy-(132-R)-phaeophytin a	Direct: All exhibited 100% inhibition  IC50: 1.96 nM, 3.11 nM, and 3.11 nM, respectively,  Post: 30% of inhibition	[107]
Anti HSV-1 and HSV-2  in vitro; Plaque reduction assay-pre-, post-; 48 h (HSV-1), 96 h (HSV-2); Vero cells; Acyclovir (IC50: 0.64 μg/mL (HSV-1), 0.80 μg/mL (HSV-2))	(1) monogalactosyl diglyceride (MGDG)  (2) digalactosyl diglyceride (DGDG)	Pre: Exhibited < 50% protective effect  Post: Exhibited 100% protective effect  Post-HSV-1: IC50: 36.00 μg/mL (MGDG), 40.00 μg/mL (DGDG), HSV-2: IC50: 41.00 μg/mL (MGDG), 43.20 μg/mL (DGDG)	[56]
Anti-dengue virus assay  in vitro; DV2 strain 16681; Real time-PCR, immunofluorescence assay; direct, pre-, post-; 5 d; Dextran sulfate (pre-), Ribavirin (post-)	(1) 132-hydroxy-(132-S)-chlorophyll b  (2) phaeophorbide A  (3) 132-hydroxy-(132-S)-phaeophytin b  (4) purpurin 18 phytyl ester	Compound 2 inhibit dengue viral 2 replication in direct and post- stages, other compounds ineffective in all stages.	[108]
(e) Anti-bacterial activity
in vitro; S. aureus, S. typhimurium; (1) Disc diffusion assay, (2) Microdilution assay; 10 mg/mL; Erythromycin (10–50 μg/μL)	(1) Stigmasterol-β-D-glucoside  (2) 3-amino-4,5-dihydroxyfuran-2(3H)-one	Compound 1: Ineffective  Compound 2 on S. aureus: 18.33 mm inhibition value, MIC: 0.3125 mg/mL  Compound 2 on S. typhimurium: 20.33 mm inhibition value, MIC: 0.625 mg/mL	[49]
(f) Anti-cancer activity
in vitro; SGC-7901 cancer cells; MTT assay; 48 h; 50, 100 and 200 μg/mL	Polysaccharide peptide complex: CNP-1-2	92.34 ± 0.94% of inhibition on cell growth at 200 μg/mL in 48 h	[59]
in vitro; A549 cells; MTT assay; 72 h	(1) 132-hydroxy-(132-S)-chlorophyll b  (2) phaeophorbide A  (3) 132-hydroxy-(132-S)-phaeophytin b  (4) purpurin 18 phytyl ester	CC50: (1) 43 μg/mL, (2) 25 μg/mL, (3) 50 μg/mL, (4) 50 μg/mL	[108]
(g) Cytotoxicity assay
in vitro; CVS assay; 72 h; Vero cells	(1) 132-hydroxy-(132-R)-phaeophytin b  (2) 132-hydroxy-(132-S)-phaeophytin a  (3) 132-hydroxy-(132-R)-phaeophytin a	Maximum concentration that is not toxic to Vero cell is: Compound 1 (5.89 μM), 2 (6.21 μM), 3 (6.21 μM)	[107]
in vitro; MTT assay; 48 h; Vero cells; 100–15000 μg/mL	(1) monogalactosyl diglyceride (MGDG)  (2) digalactosyl diglyceride (DGDG)	MGDG: CC50: 955.00 ± 7.00 μg/mL  DGDG: CC50: 922.00 ± 4.00 μg/mL	[56]