Pinus densiflora 유래의 항트롬빈 활성을 나타내는 Neolignan Glycoside의 동정 Identification of a Neolignan Glycoside from the Pine Tree, Pinus densiflora Showed Antithrombotic Activity원문보기
소나무(Pinus densiflora)의 잎의 성분을 분리하여 항트롬빈 활성을 조사하였다. 잎은 70% ethanol로 3회 추출하였으며, 그 추출물은 순차적으로 chloroform과 n-buthanol로 분획하였다. n-buthanol 분획으로부터 얻은 수용성 분획을 MPLC와 HPLC에 의해 분리하였다. 분리한 compound를 $^1H-$과 $^{13}C-NMR$로 동정한 결과 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl-3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside(a neolignan glycoside)와 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$)의 neolignan 구조임을 확인하였다. 트롬빈 저해활성은 분리된 neolignan glycoside와 icariside $E_4$을 작용하여 혈장안에서 응고시간으로 측정하였다. 그 결과, neolignan glycoside의 응고시간이 icariside $E_4$보다 4배 이상 지연하였다. 저해활성은 neolignan glycoside의 농도가 증가함에 따라 그 시간이 지연되는 것을 확인하였다. 더 나아가 지연기전을 확인하기 위해 thrombin과 순수한 fibrinogen를 반응하였다. 그 결과, 트롬빈과 순수한 피브리노겐의 작용에 의해 응고시간은 지연되었으며, 이는 neolignan glycoside가 피브린형성에 주요한 트롬빈의 활성을 직접 저해하는 것으로 사료된다.
소나무(Pinus densiflora)의 잎의 성분을 분리하여 항트롬빈 활성을 조사하였다. 잎은 70% ethanol로 3회 추출하였으며, 그 추출물은 순차적으로 chloroform과 n-buthanol로 분획하였다. n-buthanol 분획으로부터 얻은 수용성 분획을 MPLC와 HPLC에 의해 분리하였다. 분리한 compound를 $^1H-$과 $^{13}C-NMR$로 동정한 결과 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl-3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside(a neolignan glycoside)와 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$)의 neolignan 구조임을 확인하였다. 트롬빈 저해활성은 분리된 neolignan glycoside와 icariside $E_4$을 작용하여 혈장안에서 응고시간으로 측정하였다. 그 결과, neolignan glycoside의 응고시간이 icariside $E_4$보다 4배 이상 지연하였다. 저해활성은 neolignan glycoside의 농도가 증가함에 따라 그 시간이 지연되는 것을 확인하였다. 더 나아가 지연기전을 확인하기 위해 thrombin과 순수한 fibrinogen를 반응하였다. 그 결과, 트롬빈과 순수한 피브리노겐의 작용에 의해 응고시간은 지연되었으며, 이는 neolignan glycoside가 피브린형성에 주요한 트롬빈의 활성을 직접 저해하는 것으로 사료된다.
The constituents from the needles of the pine tree, Pinus densiflora, were purified and investigated for antithrombotic activity. The needles were initially extracted three times with 70% ethanol, and the extract was sequentially fractionated with chloroform and n-butanol. The aqueous layer formed a...
The constituents from the needles of the pine tree, Pinus densiflora, were purified and investigated for antithrombotic activity. The needles were initially extracted three times with 70% ethanol, and the extract was sequentially fractionated with chloroform and n-butanol. The aqueous layer formed after n-butanol fractionation was subjected to purification by medium pressure and high pressure liquid chromatography. The two neolignans, 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl -3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside (a neolignan glycoside) and 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$) were identified by $^1H$ and $^{13}C$ NMR spectra. The effect of the purified compounds, the neolignan glycoside and icariside $E_4$ on thrombin inhibition were investigated by measuring thrombin clotting time in plasma. As a result, the clotting of the neolignan glycoside was delayed four times compared to that of icariside $E_4$. In addition, an analysis of the inhibition effect by changing the concentration showed that the clotting time was delayed in accordance with an increase in the concentration of the neolignan glycoside. Furthermore, we examined the interaction of thrombin and fibrinogen to clarify the action mechanism. As a result, the delay of clotting time in the response of thrombin and pure fibrinogen may indicate that neolignan glycosides inhibit the thrombin action in a direct manner, leading to the suppression of fibrin generation.
The constituents from the needles of the pine tree, Pinus densiflora, were purified and investigated for antithrombotic activity. The needles were initially extracted three times with 70% ethanol, and the extract was sequentially fractionated with chloroform and n-butanol. The aqueous layer formed after n-butanol fractionation was subjected to purification by medium pressure and high pressure liquid chromatography. The two neolignans, 2, 3-dihydro-7-hydroxyl-3-hydroxymethyl-2-(4'-hydroxyl -3-methoxyphenyl)-5-benzofuranpropanol-3-O-${\alpha}$-rhamnopyranoside (a neolignan glycoside) and 2, 3-dihyro-3-hydroxymethyl-7-methoxy-2-(4'-hydroxyphenyl-3'-methoxy)-5-benxofuran propanol 4'-O-${\alpha}$-rhamnopyranoside (icariside $E_4$) were identified by $^1H$ and $^{13}C$ NMR spectra. The effect of the purified compounds, the neolignan glycoside and icariside $E_4$ on thrombin inhibition were investigated by measuring thrombin clotting time in plasma. As a result, the clotting of the neolignan glycoside was delayed four times compared to that of icariside $E_4$. In addition, an analysis of the inhibition effect by changing the concentration showed that the clotting time was delayed in accordance with an increase in the concentration of the neolignan glycoside. Furthermore, we examined the interaction of thrombin and fibrinogen to clarify the action mechanism. As a result, the delay of clotting time in the response of thrombin and pure fibrinogen may indicate that neolignan glycosides inhibit the thrombin action in a direct manner, leading to the suppression of fibrin generation.
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대상 데이터
1H and 13C NMR spectra were measured by Varian Unity INOVA 500 instruments (Palo Alto, CA, USA). CD3OD was used as the solvent, and the chemical shift expressed in ppm was confirmed by the solvent peak (δH 3.
densiflora needles were collected at Youngwol, Gangwondo, Korea in April 2012 and were authenticated by the Korea Environmental and Ecological Services. Plasma (bovine) and fibrinogen (Type 1-s, from bovine plasma) were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA) and thrombin (500 NIH U/ml) was supplied by Mochida Pharmaceutical Co. (Tokyo, Japan). Heparin (Sigma) was used as a control and Dade® Owren’s veronal buffer (pH 7.
The mobile phase was 43% methanol and C18-10E Shodex packed column (250×10 mm i.d., 50, 100Å) equipped with a Shodex RI-71 detector were used.
이론/모형
APTT and PT were measured using a Dade® actin and a Dade® thromboplastin C plus, respectively. Thrombin time was measured by the slight change in turbidity using the method of Born and Cross [15] and a coagulometer (KC-A1, Amelung, Japan). In a tube, 50μl 0.
성능/효과
Its relation to the anomeric proton of rhamnose on the HMBC and NOESY spectra indicated that α-rhamnose was connected to C-3a of benzofuran. These findings suggest that compound 1 was a type of neolignan containing a 2,3-dihydro-3-hydroxylmethyl-2-5-benzofuranpropanol backbone, which was previously reported in Larix leptoleps (Pinus massoniana), and Juniperus communis[21]. Optical rotation was estimated at a [α]D value of -9.
참고문헌 (26)
Choi, J. H., Kim, H. S., Jung, M. J. and Choi, J. S. 2001. (+)-Catechin, an antioxidant principle from the needles of Pinus densiflora that acts on 1,1-diphenyl-2-picrylhydrazyl radical. Nat Prod Sci 7, 1-4.
Coughlin, S. R., Faruqi, T. R., Weiss, E. J. and Shapiro, M. J. 2000. Structure-function analysis of protease-activated receptor 4 tethered ligand peptides. J Biol Chem 275, 19728-19734.
Derian, C. K., Maryanoff, B. E., Zhang, H. C. and Andrade-Gordon, P. 2003. Therapeutic potential of protease-activated receptor-1 antagonists. Expert Opin Investig Drugs 12, 209-221.
Guglielmone, H. A., Agnese, A. M., Nunez Montoya, S. C. and Cabrera, J. L. 2002. Anticoagulant effect and action mechanism of sulphated flavonoids from Flaveria videntis. Thromb Res 105, 183-188.
Jung, M. J., Choi, J. H., Chung, H. Y., Jung, J. H. and Choi, J. S. 2001. A new C-methylated flavonoid glycoside from Pinus densiflora. Fitoterapia 72, 943-945.
Jung, M. J., Chung, H. Y., Choi, J. H. C. and Choi, J. S. 2003. Antioxidant principles from the needles of red pine, Pinus densiflora. Phytother Res 17, 1064-1068.
Kahn, M. L., Zheng, Y. W., Huang, W., Bigornia, V., Zeng, D., Moff, S., Farese, R. V., Tam, C. and Coughlin, S. R. 1998. A dual thrombin receptor system for platelet activation. Nature 394, 690-694.
Kang, Y. H., Park, Y. K., Ha, T. Y. and Moon, K. D. 1996. Effects of pine needle extracts on serum and liver lipid contents in rats fed high fat diet. J Korean Soc Food Nutr 25, 367-373.
Kang, Y. H., Park, Y. K., Ha, T. Y. and Moon, K. D. 1996. Effects of pine needle extracts on enzyme activities of serum and liver, and liver morphology in rats fed high fat diet. J Korean Soc Food Nutr 25, 374-378.
Kim, T. J. 1996. Korean Resources Plants I. p. 53, Seoul National University Press, Seoul, Korea.
Kim, Y. S. and Shin, D. H. 2005. Volatile components and antibacterial effects of pine needle (Pinus densiflora S. and Z.) extracts. Food Microbiol 22, 37-45.
Kuk, J. H., Ma, S. J. and Park, K. H. 1997. Isolation and characterization of benzoic acid with antimicrobial activity from needle of Pinus densiflora. Korean J Food Sci Technol 29, 204-210.
Kuk, J. H., Ma, S. J. and Park, K. H. 1997. Isolation and characterization of cinnamic acid with antimicrobial activity from needle of Pinus densiflora. Korean J Food Sci Technol 29, 823-826.
Lee, E. 2003. Effects of powdered pine needle (Pinus densiflora seib et Zucc.) on serum and liber lipid composition and antioxidative capacity in rats fed high oxidized fat. J Korean Soc Food Sci Nutr 32, 926-930.
Lee, M. K., Sung, S. H., Lee, H. S., Cho, J. H. and Kim, Y. C. 2001. Lignan and neolignan glycosides from Ulmus davidiana var. japonica. Arch Pharm Res 24, 198-201.
Lundgren, L. N., Shen, Z. and Theander, O. 1985. The constituents of conifer neeldles, dilignol glycosides from Pinus massoniana Lamb. Act Chem Scand B 39, 241-248.
Miyase, T., Ueno, A., Takizawa, N., Kobayashi, H. and Oguchi, H. 1989. Ionone and lignan glycosides from Epimedium diphyllum. Phytochemistry 28, 3483-3485.
Morrell, C. N., Sun, N., Ikeda, M., Beique, J. C., Swaim, A. N., Mason, E., Martin, T., Thompson, L., Gozen, O., Ampagoom-ian, D., Sprengel, R., Rothstein, J., Faraday, N., Huganir, R. and Lowenstein, C. J. 2008. Glutamate mediates platelet activation through the AMPA receptor. J Exp Med 20, 575-584.
Nakanishi, T., Iida, N., Inatomi, Y., Murata, H., Inada, A., Murata, J., Lang, F. A., Iinuma, M. and Tanaka, T. 2004. Neolignan and flavonoid glycosides in Juniperus communis var. depressa. Phytochemistry 65, 207-213.
Vu, T. K., Hung, D. T., Wheaton, V. I. and Coughlin, S. R. 1991. Molecular cloning of a functional thrombin receptor reveals a novel proteolytic mechanism of receptor activation. Cell 64, 1057-1068.
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