Isolation of Flavonoids from the Fruits of Cornus kousa Burg.

Dae-Young Lee, Ha-Na Lyu, Ho-Young Kwak, Lakoon Jung, Youn-Hyung Lee,

Dae-Keun Kim, In-Sik Chung, Sung-Hoon Kim and Nam-In Baek,

Presented by Ekrima Astari

Cornus kousa
Taxonomy
Kingdom:Plantae (unranked):Angiosperms (unranked):Eudicots (unranked):Asterids Order:Cornales Family:Cornaceae Genus:Cornus Subgenus:Benthamidia Species:C. kousa

Small deciduous tree 812 m (2639 ft) tall, Native to eastern Asia. Like most dogwoods, it has opposite, simple leaves, 410 cm long. The tree is extremely showy when in bloom, but what appear to be four petaled white flowers. The blossoms appear in late spring, weeks after the tree leafs out.

Cornus kousa
Cornus kousa Burg. (Cornaceae) is a tree distributed in the mountains of Korea, China and Japan. The fruit of this plant has been used as: hemostatic agent treatment of diarrhea in Korean traditional medicine immuno-regulatory Some chemical constituents such as isoquercitrin, gallic acid, tannin, phenolics and flavonoids have been reported from the leaves of C. kousa. Also, our previous phytochemical researches on the fruits of this plant reported the presence of steroids and cytotoxic lignans

Aromadendrin 7-O--D-glucoside

Isolated Compunds
Quercetin 3-O--D-galactoside

Quercetin 3-O-a-L-rhamnoside

Quercetin 3-O-rutinoside

Kaempferol 3-O--D-galactoside Quercetin 3-O--D-glucoside Quercetin 3-O--D-glucuronide

Isolated Compunds
Triterpenes and Lignans

Lactones

Loganin

Triterpenes Glycosides

Sterols

Plant Material
The fruits of Cornus kousa Burg. (Cornaceae) Collected at the experimental farm in Kyung Hee University in August, 2006.

Experiments
Buah 10kg
Dicincang Diekstraksi 3x methanol 80% Evaporasi Etil Acetat (2L x 3) dan H2O (2L)

44g
n-hexane-EtOAc (4 : 1 2 : 1, 1.5 L) CHCl3-MeOH (15 : 113 : 110 : 1, 1.5 L) TLC

CKFE1 2.5g
CHCl3 MeOH-H2O (13 : 3 : 1, 3 L)

CKFE2 870mg
MeOH-H2O (2 : 3, 2 L)

CKFE2-1 310mg

CKFE1-2 88mg
MeOH H2O (2 : 1, 1 L) CHCl3-MeOH-H2O (12 : 3 : 1,2.5 L)

Senyawa 1 35mg

2CKFE2-1-1 (35 mg)

80% MeOH

2CKFE2-1-2 (35 mg)

Senyawa 2 35mg

Experiments
2CKFE2-1-2 (35 mg) HPLC CH3CN-H2O-TFA (50 : 50 : 0.05, vol. %) 370 nm by UV detector 2 peak: 2.97min dan 3.65min

Senyawa 3

Senyawa 4

Result and Discussion

Compound 1: Light yellow powder Compound 2: Yellow amorphous powder Compound 3: Yellow powder Compound 4: Yellow powder

Compound 1 m.p. 178- 180; UV (MeOH) max: 269, 364 nm; IR (KBr) max 3350, 1660, 1610, 1500 cm1; EI/MS m/z: 286[M+], 258, 229, 213, 184, 153,121; 1H-NMR (400 MHz, CD3OD, ) 8.06 (2H, d, J = 9.2 Hz, H-2'/6'), 6.89 (2H, d, J = 9.2 Hz, H-3/5'), 6.36 (1H, d, J = 2.0 Hz, H-8), 6.16 (1H, d, J = 2.0 Hz,H-6); H

Compound 2 m.p. 230-232oC; UV (MeOH) max: 218, 268 nm; []25 D= +16.0o (c = 1.1, MeOH); IR (KBr) max 3420, 1680 cm1; pos. FAB/MS m/z: 449, 287; 1H-NMR (400 MHz, CD3OD, ) 8.04 (2H, d, J = 8.4 Hz, H-2'/6'), 6.87 (2H, d, J = 8.4 Hz, H-3'/5'), 6.38 (1H, br s, H-8), 6.19 (1H, br s, H-6), 5.23 (1H, d, J = 7.2 Hz, H-1''), 3.18~3.70 (4H, m, H-2'', 3'', 4'', 5''), 3.71 (1H, dd, J = 12.0, 2.4 Hz, H-6a''), 3.58(1H, dd, J = 12.0, 5.2 Hz, H-6b'');

Compound 2 m.p. 230-232oC; UV (MeOH) max: 218, 268 nm; []25 D= +16.0o (c = 1.1, MeOH); IR (KBr) max 3420, 1680 cm1; pos. FAB/MS m/z: 449, 287; 1H-NMR (400 MHz, CD3OD, ) 8.04 (2H, d, J = 8.4 Hz, H-2'/6'), 6.87 (2H, d, J = 8.4 Hz, H-3'/5'), 6.38 (1H, br s, H-8), 6.19 (1H, br s, H-6), 5.23 (1H, d, J = 7.2 Hz, H-1''), 3.18~3.70 (4H, m, H-2'', 3'', 4'', 5''), 3.71 (1H, dd, J = 12.0, 2.4 Hz, H-6a''), 3.58(1H, dd, J = 12.0, 5.2 Hz, H-6b'');

Compound 3 m.p. 235-238oC; UV (MeOH) max: 207, 261 nm; []25 D= 12.5o(c = 0.9, MeOH); neg. FAB/MS m/z: 463 [M-H], 447, 431, 389, 339, 325, 301; IR (KBr) max 3400, 2919, 1656, 1606, 1508 cm1; 1H-NMR (400 MHz, CD3OD, ) 7.84 (1H, d, J = 2.4 Hz, H-2'), 7.56 (1H, dd, J = 2.4, 8.6 Hz, H-6'), 6.84 (1H, d, J = 8.6 Hz, H-5'), 6.34 (1H, d, J = 2.0 Hz, H-8), 6.16 (1H, d, J = 2.0 Hz, H-6), 5.13 (1H, d, J = 8.0 Hz, H-1''), 3.85 (dd, J = 3.2, 2.0 Hz, H-4''), 3.82 (dd, J = 8.0, 7.8 Hz, H-2''), 3.65 (dd, J = 11.0, 3.2, H-6''a), 3.56 (dd, J = 11.0, 5.5 Hz, H-6''b), 3.58 (dd, J = 7.8, 2.0 Hz, H-3'') 3.48 (m, H-5)

OH

Compound 4 m.p. 230-232 oC; UV (MeOH) max: 207, 256 nm; neg. FAB/MS m/z: 463 [M-H], 447, 423, 389, 297, 204; IR (KBr) max 3400, 2919, 1656, 1606, 1508 cm1; 1H-NMR (400 MHz, CD3OD, ) 7.70 (1H, d, J = 2.4 Hz, H-2'), 7.55 (1H, dd, J = 2.4, 8.6 Hz, H-6'), 6.85 (1H, d, J = 8.6 Hz, H-5'), 6.34 (1H, d, J = 2.0 Hz, H-8), 6.16 (1H, d, J = 2.0 Hz, H-6), 5.22 (1H, d, J = 7.2 Hz, H-1''), 3.22~3.51 (4H, m, H-2'', 3'', 4'', 5''), 3.71 (1H, dd, J = 12.0, 2.4 Hz, H-6a''), 3.58 (1H, dd, J = 12.0, 5.2 Hz, H-6b'');

OH

Result and Discussion

Compound 1: Kaempferol examined as an inhibitor of cigarette smoke-induced activation of the aryl hydrocarbon receptor and cell transformation Compound 2: Astragalin (kaempferol-3-O-Dglucopyranoside) , had cytotoxicity against four human cancer cell lines Compound 3: Hyperin (quercetin-3-O--Dgalactopyranoside) was reported to be a source of antiretroviral for AIDS therapy due to significant antiHIV-1 activity Compound 4: Isoquercitrin (quercetin-3-O--Dglucopyranoside) has also been found to be a major factor in anti-inflammatory and hepatoprotective activity All the isolated flavonoids have been reported to exhibit antioxidant activity

Shikimate Pathways

SHIKIMIC ACID CHORISMIC ACID

(+ acetogenin piece)

FLAVONOIDS

PREPHENIC ACID

PHENYL-C3 COMPOUNDS
CINNAMIC ACIDS PHENYL-C1 COMPOUNDS

ANTHRANILIC ACID

TYROSINE PHENYLALANINE

TRYPTOPHAN NEXT LECTURE

ALKALOIDS

ALKALOIDS

ALKALOIDS

Biosynthesis

Biosynthesis