KR100238110B1 - Preparation method of fraction from trichosanthes radix showing bloodmuscle relaxation - Google Patents
Preparation method of fraction from trichosanthes radix showing bloodmuscle relaxation Download PDFInfo
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
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- A61K36/185—Magnoliopsida (dicotyledons)
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Abstract
본발명은 괄루근으로부터 혈관이완능이 있는 분획을 제조하는 방법에 관한 것으로서, 더욱 상세하게는 한약재로 사용되는 괄루근(천화분)으로부터 유화성분을 알콜수용액 및 비극성용매로 추출하여 혈관이완능이 있는 괄루근 분획을 간단한 방법으로 제조하는 방법에 관한 것이다.The present invention relates to a method for producing a fraction with vasodilatation ability from the sphincter muscle, more specifically, the sphincter muscle with vasodilatability by extracting the emulsion component from the sphincter muscle (hypothelium) used as a herbal medicine with an alcoholic solution and a nonpolar solvent A method for preparing fractions in a simple manner.
Description
제1도는 본발명의 실시예 1에서 얻어진 혈관이완화합물 분획의 박층크로마토그래피(TLC)의 데이타이다.1 is data of thin layer chromatography (TLC) of the vasorelaxant fraction obtained in Example 1 of the present invention.
본발명은 괄루근으로부터 혈관이완능이 있는 분획을 제조하는 방법에 관한 것으로서, 더욱 상세하게는 한약재로 사용되는 괄루근(천화분)으로부터 혈관이완능이 있는 관루근 분획을 간단한 방법으로 제조하는 방법에 관한 것이다.The present invention relates to a method for producing a vasodilatable fraction from the sphincter muscle, and more particularly, to a method for producing a vasorelaxant capillary muscle fraction from a sphincter muscle (hypothelium) used as a medicinal herb by a simple method. .
괄루근은 동속 근연식물(박과)의 다년생 초본식물의 뿌리로서 우리나라에서는 제주도와 중북부산지 또는 수림아래에 야생으로 존재하고 있으며, 그 주요 성분으로는 스티그마스테롤(Stigmasterol), 스피나스테롤(Spinasterol), 여러지방산, 사포닌(Saponin), 폴리사카라이드(Polvsaccharide), 갈락토오즈결합렉틴(Galactose binding lectin), 트리코산틴(Trichosanthin), 트립신저해제(Trypsin inhibitor)등이 알려져 있다.Root-root roots are perennial herbaceous plants of the same genus, and they exist as wild under Jeju Island, Mid-Northern Mountain, or Forest. The main components are Steigmasterol and Spinasterol. Various fatty acids, saponins, saponins, polysaccharides, galactose binding lectins, trichosanthins, trypsin inhibitors, and the like are known.
이러한 괄루근을 탕제로 사용할 경우 동의보감 등에 나타난 한의학적 약리 작용으로는 청량, 지갈, 해열, 이노, 진혈, 구담, 당뇨병치료 등에 효과가 있는 것으로 알려져 있다.It is known that the oriental medicine pharmacological action shown in synonym and the like may be effective in cooling, jigal, antipyretic, ino, blood, gudam, and diabetes treatment.
또한 현대의학에서는 괄루근 중에서 트리코산틴을 분리, 정제하여 낙태제등에 유효생체 활성물질로 이용하고 있을 뿐만아니라 괄루근의 엑기스를 맥문동등과 복합처방에 의하여 당뇨병치료제로서 이용되기도 하고 있으나, 특이적으로 혈관이완능이 있다는 보고는 보고는 있지 않다.In modern medicine, tricoxanthin is isolated and purified from the sphincter muscle and used as an active biologically active substance for abortions. There are no reports of vascular dysfunction.
따라서 본발명의 발명자들은 괄루근으로부터 간단한 제조방법에 의하여 괄루분획을 분리 제조하고 이 분획이 혈관이완능을 가지고 있음을 알게되어 본발명을 완성하였다.Therefore, the inventors of the present invention separated the preparation of the ancestor fraction from the sphincter muscle by a simple manufacturing method, and realized that the fraction had vasodilatability, thus completing the present invention.
본발명은 괄루근으로부터 혈관이완능일 있는 괄루근분획을 간단하게 제조할 수 있는 방법을 제공하는데 그 목적이 있다.It is an object of the present invention to provide a method for the simple production of sphincter fragments with vasodilatability from the sphincter muscle.
이하, 본발명을 상세히 설명하면 다음과 같다.Hereinafter, the present invention will be described in detail.
본발명은 잘게 마쇄된 괄루근을 비극성용매로 추출, 여과하여 지질 등의 비극성물질을 제거한후 메탄올의 수용액으로 추출한후 세포잔사를 제거하고, 그 상등액을 농축하여 수용액상으로 만든후 비극성용매로 스티그마스테롤 등의 비극성 물질들을 제거한다음, 여액을 수포화부탄올로 층분리하고 증류수로 세척하여 혈관이완능이 있는 분획을 제조하는 것을 그 특징으로 한다.According to the present invention, finely crushed sphincter muscles are extracted and filtered with a nonpolar solvent to remove nonpolar substances such as lipids, and then extracted with an aqueous solution of methanol, cell residues are removed, the supernatant is concentrated to an aqueous solution, and stigma is used as a nonpolar solvent. After removing non-polar substances such as sterol, the filtrate is separated by saturated butanol and washed with distilled water to prepare a fraction having vascular relaxation ability.
또한, 본발명은 상기 방법으로 얻어진 괄루근분획을 유효성분으로 함유하는 혈관이완제를 포함한다.The present invention also includes a vasodilator containing the sphincter muscle fraction obtained by the above method as an active ingredient.
이와같은 본발명을 더욱 상세히 설명하면 다음과 같다.The present invention will be described in more detail as follows.
본 발명은 괄루근으로부터 혈관이완능이 있는 분획을 제조하는 방법에 관한 것으로서, 먼저 건조된 괄루근을 잘게 마쇄하여 비극성용매로 추출, 여과하여 지질 등의 비극성물질을 제거한후, 저급알콜의 수용액으로 추출하여 세포잔사 등을 제거하고, 그 상등액을 농축하여 수용액상으로 만든후 수용액층을 비극성용매로 층분리하여 불순물 및 잔존하는 비극성물질을 제거시킨다.The present invention relates to a method for producing a fraction with vasodilatation ability from the sphincter muscle, first grinding the dried sphincter muscle finely, extracted with a nonpolar solvent, filtered to remove nonpolar substances such as lipids, and then extracted with an aqueous solution of lower alcohol Cell residues are removed, the supernatant is concentrated to form an aqueous solution, and the aqueous layer is separated with a nonpolar solvent to remove impurities and remaining nonpolar materials.
여기서 상기 괄루근 추출시의 비극성용매로는 헥산, 클로로포름 또는 디클로로메탄 등이 사용 가능하고 이중 헥산을 사용하는 것이 환경, 취급상의 용이성 및 효율면에서 가장 바람직하며 또한, 상기 층분리시의 비극성용매로는 클로로포름, 디클로로메탄, 사염화탄소 또는 벤젠 등을 사용할 수 있는데 이중 클로로포름 또는 디클로로메탄을 사용하는 것이 극성(polarity)이 적당하여 수용액으로부터 비극성물질을 효율적으로 제거시키는 면에서 바람직하다.Herein, hexane, chloroform or dichloromethane may be used as the nonpolar solvent when extracting the sphincter, and it is most preferable to use double hexane in terms of environment, ease of handling and efficiency, and as the nonpolar solvent at the time of separating the layers. Chloroform, dichloromethane, carbon tetrachloride or benzene may be used. Of these, chloroform or dichloromethane is preferably used in view of efficiently removing the nonpolar material from the aqueous solution due to its proper polarity.
또한, 상기의 저급알콜로는 메탄올, 에탄올 또는 부탄올 등이 사용될 수 있으나, 메탄올을 사용하는 것이 경제적측면 및 수율 등의 측면에서 특히 좋으며, 이러한 저급알콜수용액의 추출에 의해 세포잔사와 가역적으로 변성된 단백질 등이 제거된다.In addition, as the lower alcohol, methanol, ethanol or butanol may be used, but it is particularly preferable to use methanol in terms of economics and yield, and the reversible denaturation with the cell residue by extraction of the lower alcohol aqueous solution. Protein and the like are removed.
다음으로 이용되는 비극성용매의 층분리에서는 스티나스테롤 등의 스테로이드와 같은 불순물 및 잔존 비극성물질이 제거되는데, 그후 얻어진 여액을 수포화된 저급알콜용매로 분리하여 저급알콜층을 모은 후 증류수로 세척하여 당류 등의 불필요한 극성물질을 다시 제거, 농축시켜 본발명의 혈관이완능이 있는 분획을 제조할 수 있게 된다.Next, in the layer separation of the non-polar solvent used, impurities and residual non-polar substances such as steroids such as stinasterol are removed. The filtrate is then separated with a saturated lower alcohol solvent to collect a lower alcohol layer and washed with distilled water. Unnecessary polar substances such as sugars are removed again and concentrated to prepare a fraction having vascular relaxation ability of the present invention.
여기서 마지막단계로 사용된 저급알콜로는 부탄올, 이소프로필알콜 또는 펜탄올 등이 사용될 수 있으나 부탄올을 사용하는 것이 수층과의 층분리 및 원하는 분획의 수율 등의 측면에서 가장 좋다.Butanol, isopropyl alcohol or pentanol may be used as the lower alcohol used as the last step, but the use of butanol is best in terms of layer separation with the aqueous layer and yield of the desired fraction.
이렇게 제조된 괄루근 분획에 대해 적출된 쥐의 대동맥을 원형으로 만들어 등력전달기를 이용하여 장력변화를 측정함으로써 혈관이완능이 확인되었다.Vascular relaxation was confirmed by making the aorta of the rat extracted from the sphincter muscle fraction thus prepared into a circular shape and measuring the tension change using an isometric force transmitter.
또 이와같이 제조된 분획은 사포닌계통의 화합물로 구성되어 있음이 확인되었다.In addition, the fraction thus prepared was confirmed to be composed of a compound of the saponin system.
따라서, 본발명은 상기와 같이 괄루근으로부터 혈관이완능이 있는 분획을 간단하게 제조할 수 있는 방법으로서, 본발명에 의해 제조된 괄루근 분획은 혈관 이완제 및 고혈압치료제의 유효성분으로 함유되어 널리 유용하게 사용될 수 있다.Therefore, the present invention is a method that can easily prepare a fraction with vascular relaxation ability from the sphincter as described above, the sphincter muscle fraction prepared by the present invention is widely used as an active ingredient of vascular relaxant and hypertension therapeutic agent Can be used.
이하, 본발명을 실시예에 의거 상세히 설명하면 다음과 같은바 본발명이 실시예에 의해 한정되는 것은 아니다.Hereinafter, the present invention will be described in detail with reference to Examples, but the present invention is not limited by the Examples.
[실시예 1]Example 1
건조된 괄루근 1kg을 잘게 마쇄하여 5l의 헥산으로 2시간씩 2회 추출한후 여과하여 헥산층을 제거시킨다음 70% 메탄올수용액 10l를 넣고 환류시키면서 4시간씩 2회 추출후 세포잔사 등을 제거한다.Finely crush 1kg of dried glutinous root, extract twice with 5l of hexane for 2 hours, remove the hexane layer by filtration, remove 10% of aqueous solution of 70% methanol solution, extract it twice under reflux for 4 hours, and remove cell residues. .
상기 여액을 감압농축하여 메탄올을 제거시킨후 클로로포름 3l씩 3회 층분리하여 클로로포름 층을 제거시킨후 수포화부탄올 3l씩 3회 반복 추출한후 부탄올 층을 모아 3l의 증류수로 세척한후 부탄올 층을 감압농축시켜 37.5g의 엑기스를 얻었다. 그 결과 얻어진 엑기스에 대해 TLC 측정결과는 제1도에 도시하였다.The filtrate was concentrated under reduced pressure to remove methanol, and the layers were separated three times by 3 l of chloroform to remove the chloroform layer. After repeated extraction of 3 l of saturated butanol three times, the butanol layers were collected and washed with 3 l of distilled water. Concentration gave 37.5 g of extract. The TLC measurement results for the resulting extract are shown in FIG.
[실시예 2]Example 2
건조된 괄루근 1kg을 잘게 마쇄하여 5l의 헥산으로 2시간씩 2회 추출한후 여과하여 헥산층을 제거시킨후 50% 메탄올수용액 10l를 넣고 환류시키면서 4시간씩 2회 추출후 세포잔사 등을 제거한다.Finely crush 1kg of dried glutinous root and extract twice with 5l of hexane for 2 hours, remove the hexane layer by filtration, add 10l of 50% methanol aqueous solution and extract twice for 4 hours while refluxing to remove cell residues. .
상기 여액을 감압농축하여 메탄올을 제거시킨후 디클로로메탄 4l씩 3회 층분리하여 디클로로메탄층을 제거시킨후 수포화부탄올 4l씩 3회 반복 추출한후 부탄올 층을 모아 4l의 증류수로 세척한후 부탄올 층을 감압 농축시켜 40.2g의 엑기스를 얻었다.The filtrate was concentrated under reduced pressure to remove methanol, and the layers were separated three times by 4 l of dichloromethane to remove the dichloromethane layer. After repeated extraction of four times of saturated butanol 4 times, the butanol layers were collected and washed with 4 l of distilled water, followed by butanol layer. The mixture was concentrated under reduced pressure to obtain 40.2 g of extract.
[실험예]Experimental Example
펜토바비탈 소듐(pentobarbital sodium)으로 마취시킨 24g, 5주령인 ICR계 쥐로부터 적출한 대동맥을 상기 실시예 1에서 얻은 괄루근엑기스를 함유한 트리스-티로드(Tris-Tyrode)용액에 담근후 5×10-6M 노르에피네프린(norepinephrine)으로 야기시킨 혈관수축을 이완시키는 정도를 등력(等力)전달기(isometric force transducer)로 측정하여 그 결과를 다음 표 1에 나타내었다.The aorta extracted from 24 g, 5 week old ICR rats anesthetized with pentobarbital sodium was immersed in Tris-Tyrode solution containing the sphincter extract obtained in Example 1 The degree of relaxation of vasoconstriction caused by × 10-6 M norepinephrine was measured with an isometric force transducer, and the results are shown in Table 1 below.
[표 1]TABLE 1
상기 표 1에서 알수 있듯이 본발명에 의하여 얻어진 괄루근분획은 적은 농도(0.01%)에서 5×10-6M의 노르에피네프린에 의해 야기된 혈관수축을 원상으로 회복시켰음을 알 수 있다.As can be seen in Table 1, the sphincter fraction obtained by the present invention can be seen that the vasoconstriction caused by norepinephrine of 5 × 10 −6 M at a low concentration (0.01%) has been restored.
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| Application Number | Priority Date | Filing Date | Title |
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| KR1019920025288A KR100238110B1 (en) | 1992-12-23 | 1992-12-23 | Preparation method of fraction from trichosanthes radix showing bloodmuscle relaxation |
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| KR1019920025288A KR100238110B1 (en) | 1992-12-23 | 1992-12-23 | Preparation method of fraction from trichosanthes radix showing bloodmuscle relaxation |
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| KR940013526A KR940013526A (en) | 1994-07-15 |
| KR100238110B1 true KR100238110B1 (en) | 2000-02-01 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100416400B1 (en) * | 2001-02-14 | 2004-01-31 | 우원홍 | Composition containing extracts from dried roots of Trichosantes kirilowii maxim for external application and skin-whitening and process for preparation thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100416400B1 (en) * | 2001-02-14 | 2004-01-31 | 우원홍 | Composition containing extracts from dried roots of Trichosantes kirilowii maxim for external application and skin-whitening and process for preparation thereof |
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