KR20160089565A - Method for preparing jicama extract inhibitig carbohydrate digestive enzyme - Google Patents
Method for preparing jicama extract inhibitig carbohydrate digestive enzyme Download PDFInfo
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- KR20160089565A KR20160089565A KR1020150008579A KR20150008579A KR20160089565A KR 20160089565 A KR20160089565 A KR 20160089565A KR 1020150008579 A KR1020150008579 A KR 1020150008579A KR 20150008579 A KR20150008579 A KR 20150008579A KR 20160089565 A KR20160089565 A KR 20160089565A
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- Prior art keywords
- extract
- extraction
- water
- present
- hicoma
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Abstract
Description
본 발명은 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법에 관한 것으로, 보다 상세하게는 세척, 건조 후 분말화한 히카마 분말에 추출용매를 가하여 초음파 분쇄하면서 50~80℃에서 4~6시간 동안 추출하는 단계를 포함하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법에 관한 것이다. More particularly, the present invention relates to a method for preparing a hycama extract having a carbohydrate digestive enzyme inhibiting effect. More particularly, the present invention relates to a method for producing Hycima extract having an effect of inhibiting carbohydrate digestion, And extracting the extracts for a period of time. The present invention also relates to a method for producing Hycima extract having a carbohydrase inhibiting effect.
당뇨병은 유전적, 환경적 요인에 의해 인슐린 분비 감소 및 저항성 등과 같이 인슐린 분비에 문제가 있거나 인슐린의 기능에 이상이 생겨 혈액 속의 포도당이 세포로 전달, 저장되지 못하고 혈액 중에 지나치게 많아져 혈당의 수치가 정상인보다 훨씬 높아지는 고혈당증(hyperglycemia) 증상을 보이는 심각한 대사성 질환이다. 당뇨병은 합병증을 포함하여 현재 우리나라 사망원인 중 5번째로 높은 것으로 알려져 있을 만큼 증가하는 추세이다. 당뇨병에 의한 다양한 합병증으로 인한 환자의 수명은 5~10년 정도 단축되는 것으로 보고되고 있다.Diabetes is caused by insulin secretion problems such as decreased insulin secretion and resistance due to genetic and environmental factors, or abnormal function of insulin, so that glucose in the blood can not be transferred to cells and stored in the blood, It is a serious metabolic disease with hyperglycemia which is much higher than normal people. Diabetes mellitus is the fifth most common cause of death in Korea, including complications. It has been reported that the life span of patients due to various complications due to diabetes is shortened by 5 to 10 years.
당뇨병 치료의 목표는 당뇨병 환자들의 혈당을 가능한 한 정상혈당으로 유지하는 것이다. 그러나 혈당의 정상 유지는 어려운 과제이며, 현재 공복 혈당뿐만 아니라 식후고혈당의 조절이 매우 강조되고 있다. 우리나라 식단의 탄수화물 중 대부분은 전분 형태로 섭취하게 된다. 전분은 α-아밀라아제 및 α-글루코시다제에 의해 소화된 후 흡수된다. 따라서 α-아밀라아제 또는 α-글루코시다제 저해활성을 가진 물질은 식후 혈당 증가를 억제하여 식후 혈당조절에 도움을 줄 수 있다. The goal of diabetes treatment is to keep blood sugar in people with diabetes as normal as possible. However, the maintenance of normal blood glucose is a difficult task. Currently, the regulation of postprandial hyperglycemia is emphasized as well as fasting glucose. Most of the carbohydrates in our diet are taken in starch form. Starch is digested by? -Amylase and? -Glucosidase and then absorbed. Therefore, a substance having an inhibitory activity of? -Amylase or? -Glucosidase may help postprandial glucose control by suppressing postprandial increase of blood glucose.
당뇨병은 진행성 질환으로 초기에는 약으로 조절가능하나 점차적으로 증상이 악화되어 결국에는 인슐린 주사를 투여받아야 한다. 그러나 시간이 경과함에 따라 점점 많은 양의 약물이 요구되고, 부작용이 많기 때문에 이를 예방하기 위해 독성은 없으면서 장기간 당뇨 환자들이 섭취하여도 될 천연물 유래 혈당 조절 기능성 식품에 대한 연구 및 상품화가 시급한 상황이다. Diabetes mellitus is a progressive disease that can be initially controlled medically, but gradually worsens and ultimately requires insulin injections. However, as time passes, increasing amounts of drugs are required, and in order to prevent this, it is urgently necessary to research and commercialize functional foods having glucose control properties derived from natural substances, which may be consumed by long-term diabetic patients without toxicity.
한편, 히카마(pachyrhizus erosus)는 멕시코 감자라고도 불리는 콩과작물로, 주로 멕시코 전역, 중미 5개국 과테말라, 온두라스, 니카라과에서 많이 생산된다. 이외에도 필리핀, 인도네시아, 중국 남부 지역 등 아시아에서도 재배된다. 최근에 우리나라에서 히카마 재배에 성공함에 따라 국내에 보급되어지는 초기단계라고 할 수 있다. On the other hand, Hachama ( pachyrhizus erosus ), also known as Mexican potatoes, is produced mainly in Mexico, Guatemala, Honduras and Nicaragua in five Central American countries. It is cultivated in Asia such as the Philippines, Indonesia and southern China. Recently, it is an early stage to be distributed in Korea due to successful cultivation of Hikoma in Korea.
또한 최근 히카마에 천연 인슐린이라고 불리는 이눌린을 많이 함유하고 있는 사실이 알려지면서 더욱 주목받고 있으며, 이외에도 식이섬유소로서 변비개선, 장질환 예방, 콜레스테롤 감소 등의 효과가 있다.In addition, recently, Hikoma has been known to contain a large amount of inulin called natural insulin, and it has become more and more remarkable. In addition, dietary fiber has effects such as improving constipation, preventing bowel disease, and decreasing cholesterol.
상기와 같은 종래기술의 문제점을 해결하고자, 본 발명은 α-글루코시다제와 같은 탄수화물 소화효소 저해 효과가 우수하며, 혈당강하 효과가 탁월한 히카마 추출물의 제조방법을 제공하는 것을 목적으로 한다.It is another object of the present invention to provide a method for producing Hycima extract which is excellent in the inhibitory effect of carbohydrate digestive enzymes such as? -Glucosidase and is excellent in blood glucose lowering effect.
또한 본 발명은 독성이 없고 장기간 복용 시에도 부작용이 없어 당뇨병 관련 질환의 예방, 개선, 치료를 위한 치료제, 식품 등에 적용하기 적합한 히카마 추출물의 제조방법을 제공하는 것을 목적으로 한다.It is another object of the present invention to provide a method for producing Hikoma extract which is suitable for application to therapeutic agents and foods for prevention, improvement and treatment of diabetes-related diseases without toxicity and no side effects even when taken for a long time.
또한 본 발명은 탄수화물 소화효소 저해 효과 및 혈당강하 효과가 우수한 당뇨병 관련 질환의 예방, 개선 또는 치료용 조성물을 제공하는 것을 목적으로 한다.Another object of the present invention is to provide a composition for preventing, ameliorating or treating diabetes-related diseases, which is excellent in the effect of inhibiting carbohydrate digestive enzymes and lowering blood glucose levels.
상기 목적을 달성하기 위하여, 본 발명은 히카마를 세척한 후 건조하는 단계; 상기 건조한 히카마를 분쇄하여 분말화하는 단계; 상기 히카마 분말에 추출용매를 가하여 초음파 분쇄하면서 50~80℃에서 4~6시간 동안 추출하는 단계; 및 상기 히카마 추출물을 감압농축하는 단계;를 포함하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법을 제공한다.In order to accomplish the above object, the present invention provides a method of manufacturing a honeycomb structure, Pulverizing and drying the dried hicamas; Adding an extraction solvent to the Hikama powder and extracting it at 50 to 80 ° C for 4 to 6 hours while ultrasonically pulverizing it; And concentrating the Hikoma extract at a reduced pressure. The present invention also provides a method for producing Hikoma extract having a carbohydrase inhibiting effect.
상기 추출용매는 물, 50% 에탄올 및 50% 메탄올 중 선택된 어느 하나 이상인 것이 바람직하다. It is preferable that the extraction solvent is at least one selected from water, 50% ethanol and 50% methanol.
특히, 상기 추출용매는 물인 것이 더욱 바람직하다.Particularly, the extraction solvent is more preferably water.
상기 초음파 분쇄는 초음파 분쇄기를 이용하여 10~60초 동안 작동시킨 후 10~20초 동안 중지시키는 동작을 반복하여 수행될 수 있다.The ultrasonic pulverization may be performed by repeating the operation for 10 to 60 seconds using the ultrasonic pulverizer and then stopping for 10 to 20 seconds.
상기 추출은 55~65℃에서 5시간 동안 추출하는 과정을 3회 반복하여 수행되는 것이 바람직하다. It is preferable that the extraction is performed by repeating the process of extracting at 55 to 65 ° C for 5 hours three times.
또한 본 발명은 전술한 방법으로 제조된 히카마 추출물을 유효성분으로 포함하는 당뇨병 관련 질환의 예방 또는 치료용 약학 조성물을 제공한다.The present invention also provides a pharmaceutical composition for the prevention or treatment of diabetes-related diseases comprising the hycima extract prepared by the above-mentioned method as an active ingredient.
상기 히카마 추출물은 조성물 총 중량에 대하여 0.01~95중량%로 포함되는 것이 바람직하다.The hicame extract is preferably contained in an amount of 0.01 to 95% by weight based on the total weight of the composition.
또한 본 발명은 전술한 방법으로 제조된 히카마 추출물을 유효성분으로 포함하는 당뇨병 관련 질환의 예방 또는 개선용 식품 조성물을 제공한다. The present invention also provides a food composition for preventing or ameliorating a diabetes-related disease comprising the hycima extract prepared as described above as an active ingredient.
본 발명에 따르면 α-글루코시다제와 같은 탄수화물 소화효소 저해 효과가 우수하며, 혈당강하 효과가 탁월한 히카마 추출물을 효율적으로 제조할 수 있으며, 이렇게 제조된 히카마 추출물은 전술한 효과와 더불어 독성이 없고 장기간 복용 시에도 부작용이 없어 당뇨병 관련 질환의 예방, 개선, 치료를 위한 치료제, 식품 등에 사용하기 적합하다. According to the present invention, it is possible to efficiently produce Hycima extract having an excellent inhibitory effect on carbohydrate digestive enzymes such as? -Glucosidase and excellent blood glucose lowering effect, and the Hycima extract thus prepared has toxicity There is no side effect even when taken for a long time, and it is suitable for use in a therapeutic agent and food for prevention, improvement and treatment of diabetes related diseases.
도 1은 본 발명의 일실시예에 따라 추출용매를 달리하여 추출한 히카마 추출물들의 α-글루코시다아제 저해활성을 나타낸 도이다.
도 2는 본 발명의 일실시예에 따라 추출온도를 20, 40, 60, 80, 100℃로 달리하여 추출한 히카마 추출물들의 α-글루코시다아제 저해활성을 나타낸 도이다.
도 3은 본 발명의 일실시예에 따라 추출온도를 55, 60, 65℃로 달리하여 추출한 히카마 추출물들의 α-글루코시다아제 저해활성을 나타낸 도이다.
도 4는 본 발명의 일실시예에 따라 추출용매로 물을 사용하고, 60℃의 추출온도에서 열수추출하여 제조한 히카마 물 추출물의 당뇨유발 생쥐에서의 식후 혈당 상승 억제 효과를 나타낸 도이다.
도 5는 본 발명의 일실시예에 따라 추출용매로 물을 사용하고, 60℃의 추출온도에서 열수추출하여 제조한 히카마 물 추출물의 정상 생쥐에서의 식후 혈당 상승 억제 효과를 나타낸 도이다.FIG. 1 is a graph showing the activity of inhibiting? -Glucosidase in hycima extracts extracted with different extraction solvents according to an embodiment of the present invention.
FIG. 2 is a graph showing the activity of inhibiting the α-glucosidase activity of Hycima extracts extracted at different extraction temperatures of 20, 40, 60, 80 and 100 ° C. according to an embodiment of the present invention.
FIG. 3 is a graph showing the activity of inhibiting the α-glucosidase activity of Hycima extracts extracted at different extraction temperatures of 55, 60, and 65 ° C. according to an embodiment of the present invention.
FIG. 4 is a graph showing an effect of suppressing postprandial blood glucose increase in diabetic mice induced by extracting Hikama water by using water as an extraction solvent and hot water extraction at an extraction temperature of 60 ° C according to an embodiment of the present invention.
FIG. 5 is a graph showing an effect of suppressing postprandial blood glucose increase in normal mice of Hikama water extract prepared by using water as an extraction solvent and hot water extraction at an extraction temperature of 60 ° C according to an embodiment of the present invention.
이하 본 발명을 상세히 설명한다. Hereinafter, the present invention will be described in detail.
본 발명자는 당뇨병 환자의 혈당을 개선할 수 있는 천연물 소재 건강기능식품을 연구하던 중, 추출용매와 추출조건에 따라 탄수화물 소화효소 저해 효과가 달라짐을 확인하고, 최대의 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 추출방법을 검증하고, 이를 토대로 본 발명을 완성하였다. The inventors of the present invention found that the inhibitory effect of carbohydrate digestion enzymes was changed according to the extraction solvent and extraction conditions while studying the natural food and health functional food which can improve the blood sugar of the diabetic patients and confirmed that the carbohydrate digestive enzyme- The present invention has been completed on the basis of this verification.
또한 본 발명은 탄수화물 소화효소 저해 효과가 탁월한 히카마 추출물을 제조함으로써 당뇨병 및 당뇨 합병증을 예방, 개선 또는 치료하는데 유용하게 이용할 수 있을 것으로 예측하였다. Further, the present invention has anticipated that the present invention can be usefully used for preventing, ameliorating or treating diabetes and diabetic complications by producing hycima extract having excellent inhibitory effect on carbohydrate digestive enzymes.
본 발명의 히카마 추출물은 특정 추출용매를 사용하여 일정한 추출조건에서 추출하여 제조할 수 있다.The Hicoma extract of the present invention can be prepared by extracting at a certain extraction condition using a specific extraction solvent.
이하 본 발명에서 설명하는 히카마 추출물은 추출, 정제의 단계에서 얻어지는 모든 추출물, 정제물, 그들의 희석액, 농축액 또는 건조물을 모두 포함하는 개념이다.The Hikoma extract described in the present invention is a concept including all the extracts obtained in the extraction and purification steps, the purified product thereof, the diluted solution thereof, the concentrate or the dried product.
먼저, 히카마를 세척한 후 건조한다. First, wash the hicamas and dry them.
상기 세척은 정제수를 사용하여 수행될 수 있으며, 건조에 앞서 히카마를 절단하는 단계를 추가로 더 수행할 수 있다.The washing may be carried out using purified water, and a further step of cutting the hicamas may be further performed before drying.
상기 히카마는 통상의 절단기를 이용하여 0.1~1㎝의 두께로 절단할 수 있으며, 상기 절단단계를 통해 건조시간을 단축할 수 있는 효과가 있다.The hicahas can be cut to a thickness of 0.1 to 1 cm using a conventional cutter, and the drying time can be shortened through the cutting step.
이후 절단된 히카마는 50~70℃, 바람직하게는 60℃에서 약 1시간 동안 건조한다. The cut hicamas are then dried at 50-70 째 C, preferably at 60 째 C for about 1 hour.
상기 건조된 히카마는 통상의 분쇄기를 이용하여 히카마의 입자가 100~150메쉬(mesh)의 크기가 되도록 분쇄한다. 이때, 히카마 입자의 크기가 상기와 같은 경우 최종 얻어지는 히카마 추출물의 탄수화물 소화효소 저해 효과를 더욱 향상시킬 수 있어 바람직하다.The dried hicama is pulverized to a size of 100 to 150 mesh using a conventional pulverizer. At this time, when the size of the hicamas is in the above range, the effect of inhibiting the carbohydrate enzyme of the finally obtained Hicoma extract can be further improved, which is preferable.
상기와 같이 분말화된 히카마 분말에는 추출용매를 가하여 초음파 분쇄하면서 추출하는 단계를 수행한다.The powdered Hicoma powder is subjected to extraction with an extraction solvent while ultrasonically pulverizing.
통상 히카마 추출물을 제조하기 위하여는 다양한 추출용매와 추출방법이 적용될 수 있으나, 본 발명에서는 탄수화물 소화효소 저해 효과가 탁월한 히카마 추출물을 제조하기 위하여 특정 추출용매를 사용하는 것이 바람직하다. 상기 추출용매로는 물, 50% 에탄올 및 50% 메탄올 중 선택된 어느 하나 이상의 용매를 사용하는 것이 바람직하며, 특히 물을 사용하는 것이 최종 얻어진 히카마 추출물의 탄수화물 소화효소 저해 효과 향상에 있어 더욱 바람직하다. 또한 상기 추출용매는 히카마 분말 중량의 10배 정도의 양으로 첨가되는 것이 바람직하다. In order to produce Hikoma extract, various extraction solvents and extraction methods can be applied. However, in the present invention, it is preferable to use a specific extraction solvent in order to produce Hikoma extract having an excellent inhibitory effect on carbohydrate digestion. As the extraction solvent, it is preferable to use at least one solvent selected from among water, 50% ethanol and 50% methanol, and it is more preferable to use water, in particular, to improve the carbohydrate digestion inhibiting effect of the finally obtained Hicoma extract . It is also preferable that the extraction solvent is added in an amount of about 10 times the weight of the hycake powder.
상기 히카마 추출물의 추출 시에는 초음파 분쇄하면서 추출하는 것이 바람직하다.When extracting the Hicoma extract, it is preferable to carry out extraction while ultrasonically pulverizing.
상기 초음파 분쇄는 통상의 초음파 분쇄기(sonicator)를 이용하여 수행할 수 있으며, 바람직하게는 10~300초 동안 작동시킨 후 10~120초 동안 중지시키는 동작, 더욱 바람직하게는 10~60초 동안 작동시킨 후 10~20초 동안 중지시키는 동작을 반복하여 짧게 끊어 여러번에 나누어 분쇄시키는 것이 추출 중 열발생에 의한 히카마 추출물에 함유되는 성분의 변성을 방지할 수 있어 더욱 좋다.The ultrasonic pulverization can be performed using a conventional ultrasonic sonicator, preferably 10 to 300 seconds and then 10 to 120 seconds, more preferably 10 to 60 seconds, It is more preferable that the operation of stopping for 10 to 20 seconds is repeated to break up the pulverized product several times and pulverize the product several times to prevent degeneration of the component contained in the extract of Hikoma by heat generation during extraction.
상기와 같이 히카마 분말은 초음파 분쇄하면서 추출하는데, 이때 추출온도는 50~80℃인 것이 바람직하며, 더욱 바람직하게는 55~65℃인 것이 바람직하다. 상기 추출 온도가 50℃ 미만일 경우에는 inulase 효소가 존재하여 inulin이 분해될 수 있으며, 80℃를 초과할 경우에는 히카마의 유효성분이 변성되어 히카마 추출물의 효용성이 떨어질 수 있다. 특히, 상기 추출온도 범위가 55~65℃인 경우에는 inulase 효소가 불활성화되어 inulin이 많이 남게 되어 히카마 추출물의 효용성이 증가되어 더욱 바람직하다.As described above, the Hicaman powder is extracted while pulverizing by ultrasonic waves, and the extraction temperature is preferably 50 to 80 ° C, more preferably 55 to 65 ° C. When the extraction temperature is lower than 50 ° C, the inulinase may be present to decompose the inulin. If the extraction temperature is higher than 80 ° C, the efficacy of the hycima may be denatured and the efficacy of the hycima extract may be deteriorated. Particularly, when the extraction temperature is in the range of 55 to 65 ° C, the inulase enzyme is inactivated to leave a large amount of inulin, thus increasing the utility of the hycima extract.
또한 상기 추출시간은 4~6시간 동안, 바람직하게는 5시간 동안 수행되는 것이 바람직하며, 상기 추출시간이 4시간 미만일 경우에는 히카마의 수용성 식이섬유 및 유효성분들이 충분히 용출되지 않을 수 있으며, 6시간을 초과할 경우에는 유효성분의 변성으로 인해 색상이 좋지 못하고 쓴맛이 날 수 있다.The extraction time is preferably 4 to 6 hours, preferably 5 hours. If the extraction time is less than 4 hours, the water soluble dietary fibers and active ingredients of Hikoma may not be sufficiently eluted, and 6 If the time is exceeded, the color of the active ingredient may be poor due to denaturation and bitter taste may result.
특히, 상기 히카마 추출물의 추출은 초음파 분쇄하면서 55~65℃에서 5시간 동안 추출하는 과정을 3회 반복하여 추출하는 것이 수용성 식이 섬유소, 이눌린 등이 히카마로부터 충분히 용출되어 보다 탄수화물 소화효소 저해효과가 높은 추출물을 얻을 수 있어 더욱 바람직하다.In particular, the extraction of Hikoma extract is carried out by repeating the extraction process for 5 hours at 55 to 65 ° C while ultrasonically pulverizing it three times repeatedly, so that water-soluble dietary fiber, inulin and the like are sufficiently eluted from the hycamma to inhibit the carbohydrate digestive enzyme Can be obtained.
상기 히카마 추출물은 열수추출법, 냉침추출법, 환류냉각추출법, 용매추출법, 수증기증류법, 초음파추출법, 용출법, 압착법 등의 방법이 사용될 수 있으며, 특히 열수추출법을 사용하는 것이 더욱 바람직하다. As the Hicoma extract, methods such as a hot water extraction method, a cold extraction extraction method, a reflux cooling extraction method, a solvent extraction method, a steam distillation method, an ultrasonic extraction method, a leaching method and a pressing method may be used. More preferably, a hot water extraction method is used.
상기 추출완료 후에는 추출액을 여과하여 원심분리한 다음, 상기 여과액을 감압농축하여 히카마 추출물을 얻을 수 있으며, 또한 추가로 용매를 증발, 분무건조 또는 동결건조할 수도 있다.After completion of the extraction, the extract is filtered and centrifuged, and the filtrate is concentrated under reduced pressure to obtain a Hikoma extract. Further, the solvent may be further evaporated, spray dried or lyophilized.
상기와 같은 방법으로 제조된 본 발명의 히카마 추출물은 전술한 바와 같은 추출조건 하에서 초음파 분쇄하면서 추출하는 방법 이외의 통상 알려진 다른 추출방법에 의해 실시하더라도 전술한 본 발명의 추출조건을 충족시키지 않을 경우에는 본 발명과 같은 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물을 제조할 수 없다. The Hicoma extract of the present invention produced by the above-described method may be carried out by other known extraction methods other than the method of extracting under ultrasonic pulverization under the extraction conditions described above, but if the extraction conditions of the present invention are not satisfied , It is not possible to produce Hikoma extract having the carbohydratease inhibiting effect as in the present invention.
상기와 같이 탄수화물 소화효소 저해 효과가 우수한 본 발명의 히카마 추출물은 추출단계에서 얻어지는 모든 추출물, 정제물, 그들의 희석액, 농축액 또는 건조물의 형태로 이용될 수도 있다.The Hicoma extract of the present invention, which is excellent in the effect of inhibiting carbohydrate digestive enzymes as described above, may be used in the form of all extracts, purified products, diluted solutions, concentrates or dried products obtained in the extraction step.
일반적으로 동일한 성분을 추출한다 하더라도 어떠한 물질로부터 추출해내는지와, 사용되는 추출용매나, 추출온도 및 추출시간, 추출방법에 따라 추출효율이 각각 상이하게 나타나며, 본 발명에서 히카마 추출물의 제조 시 기존 알려진 추출용매, 추출온도, 추출시간과 추출방법을 적용하였다 하더라도 본 발명과 같이 탄수화물 소화효소 저해 효과가 탁월한 히카마 추출물을 얻기 위한 최적의 추출조건에 대한 연구는 이전까지는 전무하였으며, 어디에도 찾아볼 수 없는 기술이므로, 본 발명은 전혀 예기치 못한 기술임은 자명한 것이다.Generally, even if the same components are extracted, extraction efficiency varies according to the extraction solvent used, extraction temperature, extraction time, and extraction method. In the present invention, Even if the extraction solvent, the extraction temperature, the extraction time and the extraction method are applied, the optimum extraction conditions for obtaining the Hycima extract having the excellent inhibitory effect on the carbohydrate digestive enzymes as in the present invention have not been studied until now Technology, it is obvious that the present invention is a completely unexpected technique.
따라서, 상기와 같은 본 발명의 추출방법에 따르면 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물을 보다 간단하고 경제적인 방법으로 단시간에 효율적으로 추출할 수 있다. Therefore, according to the extraction method of the present invention, it is possible to efficiently extract Hicoma extract having the inhibitory effect of carbohydrate digestion in a short time by a simple and economical method.
뿐만 아니라, 본 발명의 제조방법에 따라 추출된 히카마 추출물은 α-글루코시다제와 같은 탄수화물 소화효소 저해 효과가 우수하며, 혈당강하 효과가 탁월한 히카마 추출물을 효율적으로 제조할 수 있으며, 이렇게 제조된 히카마 추출물은 전술한 효과와 더불어 독성이 없고 장기간 복용 시에도 부작용이 없어 당뇨병 관련 질환의 예방, 개선, 치료를 위한 약학, 식품 등의 분야에 유용히 적용할 수 있다. In addition, the Hycima extract extracted according to the production method of the present invention is excellent in the inhibitory effect of carbohydrate digestive enzymes such as? -Glucosidase, efficiently produces Hycima extract having excellent blood glucose lowering effect, The Hikoma extract has no toxicity and has no side effects even when it is taken for a long period of time. Therefore, it can be effectively applied to the fields of pharmacy and food for the prevention, improvement and treatment of diabetes related diseases.
상기 당뇨병 관련 질환은 당뇨병 및 당뇨 합병증으로, 예를 들어 당뇨병, 당뇨병성 백내장, 신장병, 말초신경장애, 고지혈증 등일 수 있으며, 이에 제한되는 것은 아니다.The diabetes related diseases include diabetes and diabetic complications, for example, diabetes, diabetic cataract, nephropathy, peripheral neuropathy, hyperlipemia, and the like, but are not limited thereto.
이에 본 발명은 히카마 추출물을 유효성분으로 포함하는 당뇨병 관련 질환 예방 또는 치료용 약학 조성물을 제공한다.Accordingly, the present invention provides a pharmaceutical composition for preventing or treating diabetes-related diseases comprising hiccama extract as an active ingredient.
상기 히카마 추출물은 약학 조성물 총 중량에 대하여 0.01~95중량%로 포함되는 것이 바람직하며, 더욱 바람직하게는 1~80중량%로 포함되는 것이다. 그 함량이 0.01중량% 미만일 경우에는 복용의 효율성이 떨어질 수 있으며, 95중량%를 초과할 경우에는 제형화에 어려움이 있을 수 있다.The hicame extract is preferably contained in an amount of 0.01 to 95% by weight, more preferably 1 to 80% by weight, based on the total weight of the pharmaceutical composition. If the content is less than 0.01% by weight, the efficiency of taking may be poor. If the content is more than 95% by weight, formulation may be difficult.
본 발명의 조성물은 약학 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다. 또한 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 당해 기술 분야에 알려진 적합한 제제는 문헌(Remington's Pharmaceutical Science, 최근, Mack Publishing Company, Easton PA)에 개시되어 있는 것을 사용하는 것이 바람직하다. 포함될 수 있는 담체, 부형제 및 희석제로는 락토오스, 덱스트로오스, 수크로오스, 소르비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로오스, 메틸 셀룰로오스, 미정질 셀룰로오스, 폴리비닐 피롤리돈, 물, 메틸히드록시 벤조에이트, 프로필히드록시 벤조에이트, 탈크, 마그네슘 스테아레이트, 광물유 등이 있다. 상기 조성물을 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다. 경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함되며, 이러한 고형제제는 상기 조성물에 적어도 하나 이상의 부형제, 예를 들면 전분, 칼슘 카보네이트(calcium carbonate), 수크로오스, 락토오스, 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순 희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다. 비경구 투여를 위한 제제에는 멸균된 수용액, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제 등이 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다. The compositions of the present invention may further comprise suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In addition, it can be formulated in the form of powders, granules, tablets, capsules, suspensions, emulsions, oral preparations such as syrups and aerosols, external preparations, suppositories and sterilized injection solutions according to a conventional method. Suitable formulations known in the art are preferably those as disclosed in Remington ' s Pharmaceutical Science, recently, Mack Publishing Company, Easton PA. Examples of carriers, excipients and diluents which may be included include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methylcellulose, Cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil and the like. When the composition is formulated, it is prepared using a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient usually used. Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient such as starch, calcium carbonate, sucrose, lactose, Gelatin and the like. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, and syrups. In addition to water and liquid paraffin, simple diluents commonly used, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included . Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of the suppository base include witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin and the like.
본 발명에서 사용되는 용어 "투여"는 임의의 적절한 방법으로 개체에게 소정의 본 발명의 조성물을 제공하는 것을 의미한다.The term "administering" as used herein is meant to provide any desired composition of the invention to a subject in any suitable manner.
본 발명은 약학 조성물은 연구자, 수의사, 의사 또는 기타 임상에 의해 생각되는 조직계, 동물 또는 인간에서 생물학적 또는 의학적 반응을 유도하는 유효 성분 또는 약학적 조성물의 양, 즉 치료되는 질환 또는 장애의 증상의 완화를 유도하는 양인 치료상 유효량으로 투여할 수 있다. 본 발명의 약학 조성물에 대한 치료상 유효 투여량 및 투여횟수는 원하는 효과에 따라 변화될 것임은 당업자에게 자명하다. 그러므로, 투여될 최적의 투여량은 당업자에 의해 쉽게 결정될 수 있으며, 질환의 종류, 질환의 중증도, 조성물에 함유된 유효성분 및 다른 성분의 함량, 제형의 종류, 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여시간, 투여 경로 및 조성물의 분비율, 치료기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있다. 바람직한 효과를 위해서, 본 발명의 약학 조성물은 1~10,000㎎/㎏/day, 바람직하게는 1~200㎎/㎏/day 내지 의 양으로 투여할 수 있으며, 하루에 한번 투여할 수도 있고, 수 회에 나누어 투여할 수도 있다. The present invention relates to pharmaceutical compositions comprising an amount of an active ingredient or pharmaceutical composition that induces a biological or medical response in a tissue system, animal or human, as contemplated by a researcher, veterinarian, physician or other clinician, RTI ID = 0.0 > effective < / RTI > amount. It will be apparent to those skilled in the art that the therapeutically effective dose and frequency of administration for the pharmaceutical compositions of the present invention will vary with the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. For a desired effect, the pharmaceutical composition of the present invention may be administered in an amount of 1 to 10,000 mg / kg / day, preferably 1 to 200 mg / kg / day, or may be administered once a day, . ≪ / RTI >
본 발명의 약학 조성물은 개체에게 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁 내 경막 또는 뇌혈관 내 주사에 의해 투여될 수 있다. The pharmaceutical compositions of the present invention can be administered to a subject in a variety of routes. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine dural or intracerebral injection.
또한 본 발명의 조성물은 당뇨병 관련 질환의 예방 또는 치료를 위하여 단독으로, 또는 수술, 방사선 치료, 호르몬 치료, 화학 치료 또는 생물학적 반응 조절제를 사용하는 방법들과 병용하여 사용할 수 있다.In addition, the composition of the present invention can be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy or biological response modifiers for the prevention or treatment of diabetes related diseases.
또한 본 발명은 히카마 추출물을 유효성분으로 포함하는 당뇨병 관련 질환 예방 또는 개선용 식품 조성물을 제공한다. 본 발명의 히카마 추출물이 식품 첨가물로 사용할 경우, 상기 히카마 추출물을 그대로 첨가하거나, 다른 식품 또는 식품 성분과 함께 혼합하여 사용되는 등 통상적인 방법에 따라 적절하게 사용될 수 있다. The present invention also provides a food composition for preventing or ameliorating diabetes-related diseases comprising hiccama extract as an active ingredient. When the Hicoma extract of the present invention is used as a food additive, it may be suitably used according to a conventional method such as adding the Hicoma extract directly or mixing with another food or food ingredient.
또한 상기 유효성분인 히카마 추출물의 혼합양은 사용 목적(예방, 건강 또는 치료적 처치)에 따라 적합하게 변경될 수 있음은 물론이며, 상기 상기 히카마 추출물은 식품 조성물 총 중량에 대하여 0.01~95중량%로 포함되는 것이 바람직하며, 더욱 바람직하게는 1~80중량%로 포함되는 것이다. 그 함량이 0.01중량% 미만일 경우에는 복용의 효율성이 떨어질 수 있으며, 95중량%를 초과할 경우에는 제형화에 어려움이 있을 수 있다.In addition, it is a matter of course that the mixing amount of the Hicoma extract as the active ingredient may be suitably changed according to the purpose of use (prevention, health or therapeutic treatment), and the Hicoma extract is preferably 0.01 to 95 wt% %, And more preferably from 1 to 80 wt%. If the content is less than 0.01% by weight, the efficiency of taking may be poor. If the content is more than 95% by weight, formulation may be difficult.
구체적인 예로, 식품 또는 음료의 제조 시에는 본 발명의 히카마 추출물은 원료에 대하여 15중량% 이하, 바람직하게는 10중량% 이하의 양으로 첨가되는 것이다. 그러나 건강 및 위생을 목적으로 하거나 또는 건강 조절을 목적으로 하여 장기간 섭취할 경우에는 상기 범위 이하의 양으로 첨가될 수 있으며, 안전성 면에서 아무런 문제가 없기 때문에 유효성분은 상기 범위 이상의 양으로도 사용될 수 있다. As a specific example, the Hikoma extract of the present invention is added in an amount of not more than 15% by weight, preferably not more than 10% by weight based on the raw material. However, when it is intended for health and hygiene purposes or for the purpose of controlling health, it can be added in an amount below the above range, and there is no problem in terms of safety. Therefore, the active ingredient can be used in an amount exceeding the above range have.
상기 식품의 종류에는 특별한 제한은 없다. 본 발명의 히카마 추출물을 첨가할 수 있는 식품의 예로는 육류, 소시지, 빵, 초콜릿, 캔디류, 스낵류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 수프, 음료수, 차, 드링크제, 알코올 음료, 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강식품을 모두 포함한다.There is no particular limitation on the kind of the food. Examples of foods to which the hiccama extract of the present invention can be added include meat products, sausages, breads, chocolate, candies, snacks, confectionery, pizza, ramen noodles, other noodles, dairy products including ice cream, various soups, Tea, a drink, an alcoholic beverage, a vitamin complex, and the like.
본 발명의 식품 조성물이 음료로 제조될 경우 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등의 추가 성분을 포함할 수 있다. 상기 천연 탄수화물로는 포도당, 과당 등의 모노사카라이드; 말토오스, 수크로오스 등의 디사카라이드; 덱스트린, 사이클로덱스트린 등의 천연 감미제나 사카린, 아스파르탐 등의 합성 감미제 등이 사용될 수 있다. 상기 천연 탄수화물은 본 발명의 식품 조성물 총 중량에 대하여 0.01~10중량%, 바람직하게는 0.01~0.1중량%로 포함되는 것이다.When the food composition of the present invention is prepared as a beverage, it may contain additional ingredients such as various flavors or natural carbohydrates such as ordinary beverages. Examples of the natural carbohydrate include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame, and the like. The natural carbohydrate is contained in an amount of 0.01 to 10% by weight, preferably 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.
상기 외에 본 발명의 식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알코올, 탄산음료에 사용되는 탄산화제 등을 포함할 수 있다. 뿐만 아니라, 본 발명의 조성물은 천연 과일주스, 과일주스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 상기의 첨가제 비율은 크게 제한되지는 않으나, 본 발명의 식품 조성물 총 중량에 대하여 0.01~0.1중량% 범위내로 포함되는 것이 좋다. In addition to the above, the food composition of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid and salts thereof, alginic acid and its salts, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, , Carbonating agents used in carbonated beverages, and the like. In addition, the compositions of the present invention may include flesh for the production of natural fruit juices, fruit juice drinks and vegetable drinks. These components may be used independently or in combination. The proportion of the above additives is not limited to a great extent, but may be in the range of 0.01 to 0.1% by weight based on the total weight of the food composition of the present invention.
이하에서는 실시예를 들어 본 발명에 관하여 더욱 상세하게 설명할 것이나. 이들 실시예는 단지 설명의 목적을 위한 것으로 본 발명의 보호 범위를 제한하고자 하는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to examples. These embodiments are for purposes of illustration only and are not intended to limit the scope of protection of the present invention.
실시예 1. 히카마 물 추출물 제조Example 1. Preparation of Hikama water extract
히카마를 정제수로 세척한 후, 0.1~1㎝ 두께로 절단하여 60℃에서 약 1시간 동안 건조하였다. 상기 건조된 히카마를 분쇄기를 이용하여 100-150 메쉬(mesh)의 크기로 분쇄하여 분말화하였다. 상기 히카마 분말 20g에 물 200mL를 넣고 하루 동안 침지시킨 후 초음파분쇄기(sonicator)를 이용하여 60℃에서 5시간씩 3번 반복하여 열수추출하였다. 상기 추출물은 여과지(Whatman No.1)를 이용하여 여과하고, 450ㅧg, 30분간 원심분리(MICRO 17R, Hanil)하였다. 이렇게 분리된 상등액을 35℃에서 감압농축하여 히카마 추출물을 제조하였다. Hikama was washed with purified water, cut to a thickness of 0.1 to 1 cm, and dried at 60 DEG C for about 1 hour. The dried hicahas were pulverized to a size of 100-150 mesh using a pulverizer and pulverized. 20 g of the Hikama powder was immersed in 200 mL of water for one day and then subjected to hot water extraction using an ultrasonic sonicator at 60 ° C for 5 hours three times. The extract was filtered using a filter paper (Whatman No. 1) and centrifuged at 450 ㅧ g for 30 minutes (MICRO 17R, Hanil). The supernatant thus separated was concentrated under reduced pressure at 35 DEG C to prepare Hikoma extract.
실시예 2. 히카마 50% 에탄올 추출물 제조Example 2. Preparation of 50% Ethanol Extract of Hikama
상기 실시예 1에서 추출용매로 물을 대신하여 50% 에탄올을 사용하여 추출한 것을 제외하고는 상기 실시예 1과 동일하게 실시하였다.The procedure of Example 1 was repeated, except that 50% ethanol was used instead of water as the extraction solvent in Example 1 above.
실시예 3. 히카마 50% 메탄올 추출물 제조Example 3. Preparation of
상기 실시예 1에서 추출용매로 물을 대신하여 50% 메탄올을 사용하여 추출한 것을 제외하고는 상기 실시예 1과 동일하게 실시하였다.The procedure of Example 1 was repeated, except that 50% methanol was used instead of water as the extraction solvent in Example 1.
실험예 1. 추출용매에 따른 히카마 추출물의 α-글루코시다아제 저해활성Experimental Example 1. [alpha] -G glucosidase inhibitory activity of Hicoma extract according to extraction solvent
본 실험에서는 상기 실시예 1 내지 3에서 제조한 히카마 추출물의 다당류가 단당류로 분해되는 경로의 억제능, 즉 탄수화물 소화효소 저해효과를 알아보기 위하여, 다음과 같이 실험하였다. In this experiment, the inhibition of the pathway of the degradation of polysaccharide into monosaccharides of the hycima extract prepared in Examples 1 to 3, that is, the carbohydrate digestion inhibiting effect, was tested as follows.
2g/L BSA(bovine serum albumin), 0.2g/L NaN3와 함께 효소(0.7U)를 용해시켜 효소 용액을 준비하였다. p-니트로페닐-α-D-글루코피라노사이드(p-nitrophenyl-α-D-glucopyranoside)를 PBS에 5mM 농도로 용해시켜 기질용액을 만든 후, 효소용액 0.5mL에 상기 실시예 1 내지 3에서 제조한 히카마 추출물을 각각 0.1mL씩 넣은 후, 405㎚에서 흡광도를 측정하였다. 반응 전 흡광도를 측정한 후 5분간 실온에 방치하고, 기질용액 0.5mL를 넣고 5분간 반응시킨 후 다시 흡광도를 측정하였다. 이때 양성 대조구로는 당뇨병 치료제인 아카보즈(acabose)를 사용하였으며, 그 결과는 도 1에 나타내었다.The enzyme solution was prepared by dissolving the enzyme (0.7 U) with 2 g / L bovine serum albumin (BSA) and 0.2 g / L NaN 3 . A substrate solution was prepared by dissolving p-nitrophenyl-α-D-glucopyranoside in PBS at a concentration of 5 mM, and then 0.5 ml of the enzyme solution was added to each of the above- 0.1 ml of each of the Hikoma extracts prepared was added thereto, and the absorbance was measured at 405 nm. The absorbance before the reaction was measured, and the solution was allowed to stand at room temperature for 5 minutes. 0.5 mL of the substrate solution was added, and the reaction was allowed to proceed for 5 minutes. At this time, acabose, a treatment for diabetes, was used as a positive control, and the results are shown in Fig.
도 1에 나타낸 바와 같이, 본 발명에 따라 제조한 상기 실시예 1 내지 3의 히카마 추출물을 0.1㎎/mL의 농도로 사용한 경우, 히카마 물 추출물이 57.14%로 탄수화물 소화효소 저해효과가 가장 높게 나타났으며, 히카마 50% 에탄올 추출물은 26.09%, 히카마 50% 메탄올 추출물은 13.36%의 저해효과를 나타냄을 확인할 수 있었다(p<0.05). 또한 양성 대조구로 사용한 아카보즈의 경우 64.84%의 저해 효과를 나타내었으며, 본 발명에 따라 제조한 히카마 물 추출물은 아카보즈와 유사한 정도의 탄수화물 소화효소 저해효과를 나타냄을 알 수 있었으며, 이같은 결과로부터 본 발명에 따라 제조한 히카마 물 추출물의 경우 탄수화물 소화효소 저해효과를 통한 혈당강하 효과가 우수하여 당뇨병 관련 질환의 예방, 개선, 치료에 사용하기 적합할 것임을 예측할 수 있었다. As shown in Fig. 1, when the Hikoma extract of Examples 1 to 3 prepared according to the present invention was used at a concentration of 0.1 mg / mL, the Hikama water extract had 57.14% of the highest inhibitory effect on carbohydrase (50% ethanol extract, 26.3%) and 50% methanol extract of Hikoma showed 13.36% inhibition (p <0.05), respectively. In addition, the inhibition effect of the acarbose used as the positive control was 64.84%, and the Hikama water extract prepared according to the present invention showed a similar effect to the carbohydrate digestive enzyme as the acarbose. The Hikama water extract prepared according to the present invention is excellent in the blood glucose lowering effect through the inhibition of carbohydrate digestive enzymes, and thus it can be predicted that it is suitable for use in the prevention, improvement and treatment of diabetes related diseases.
실시예 4. 추출온도에 따른 히카마 물 추출물 제조Example 4. Preparation of Hikama water extract according to extraction temperature
히카마를 정제수로 세척한 후, 0.1~1㎝ 두께로 절단하여 60℃에서 약 1시간 동안 건조하였다. 상기 건조된 히카마를 분쇄기를 이용하여 100-150 메쉬의 크기로 분쇄하여 분말화하였다. 상기 히카마 분말 20g에 물 200mL를 넣고 하루 동안 침지시킨 후 초음파분쇄기(sonicator)를 이용하여 20, 40, 60, 80, 100℃에서 각각 5시간씩 3번 반복하여 열수추출하였다. 상기 추출물은 여과지(Whatman No.1)를 이용하여 여과하고, 450×g, 30분간 원심분리(MICRO 17R, Hanil)하였다. 이렇게 분리된 상등액을 35℃에서 감압농축하여 히카마 물 추출물을 제조하였다. Hikama was washed with purified water, cut to a thickness of 0.1 to 1 cm, and dried at 60 DEG C for about 1 hour. The dried hicahar was pulverized to a size of 100-150 mesh using a pulverizer and pulverized. 20 g of Hycamase powder was immersed in 200 mL of water for one day and then subjected to hot water extraction using an ultrasonic sonicator three times at 20, 40, 60, 80, and 100 캜 for 5 hours each. The extract was filtered using filter paper (Whatman No. 1) and centrifuged at 450 xg for 30 minutes (MICRO 17R, Hanil). The supernatant thus separated was concentrated under reduced pressure at 35 DEG C to prepare Hikama water extract.
실험예 2. 추출온도에 따른 히카마 물 추출물의 α-글루코시다아제 저해활성Experimental Example 2:? -Glucosidase inhibitory activity of Hikama water extract according to extraction temperature
본 실험에서는 상기 실시예 4에서 추출온도를 달리하여 제조한 히카마 물 추출물들의 탄수화물 소화효소 저해효과를 알아보기 위하여, 다음과 같이 실험하였다. In this experiment, the carbohydrate digestion inhibitory effect of Hikama water extracts prepared at different extraction temperatures in Example 4 was tested as follows.
2g/L BSA(bovine serum albumin), 0.2g/L NaN3와 함께 효소(0.7U)를 용해시켜 효소 용액을 준비하였다. p-니트로페닐-α-D-글루코피라노사이드(p-nitrophenyl-α-D-glucopyranoside)를 PBS에 5mM 농도로 용해시켜 기질용액을 만든 후, 효소용액 0.5mL에 상기 실시예 4에서 제조한 히카마 물 추출물을 각각 0.1mL씩 넣은 후, 405㎚에서 흡광도를 측정하였다. 반응 전 흡광도를 측정한 후 5분간 실온에 방치하고, 기질용액 0.5mL를 넣고 5분간 반응시킨 후 다시 흡광도를 측정하였다. 그 결과는 도 2에 나타내었다.The enzyme solution was prepared by dissolving the enzyme (0.7 U) with 2 g / L bovine serum albumin (BSA) and 0.2 g / L NaN 3 . A substrate solution was prepared by dissolving p-nitrophenyl-α-D-glucopyranoside in PBS at a concentration of 5 mM. To the enzyme solution, 0.5 mL of the solution prepared in Example 4 0.1 ml each of Hikama water extract was added thereto, and the absorbance was measured at 405 nm. The absorbance before the reaction was measured, and the solution was allowed to stand at room temperature for 5 minutes. 0.5 mL of the substrate solution was added, and the reaction was allowed to proceed for 5 minutes. The results are shown in Fig.
도 2에 나타낸 바와 같이, 추출온도 조건을 달리하여 제조한 상기 실시예 4의 히카마 물 추출물을 0.1㎎/mL의 농도로 사용한 경우, 60℃의 추출온도에서 추출한 히카마 물 추출물이 57.14%로 탄수화물 소화효소 저해효과가 가장 높게 나타났으며, 20℃에서 추출한 추출물은 23.42%, 40℃에서 추출한 추출물은 27.34%, 80℃에서 추출한 추출물은 41.75%, 100℃에서 추출한 추출물은 34.32%로 각각 나타남을 확인할 수 있었다(p<0.05). As shown in FIG. 2, when the Hikama water extract of Example 4 prepared at different extraction temperature conditions was used at a concentration of 0.1 mg / mL, the Hikama water extract extracted at an extraction temperature of 60 ° C was 57.14% The carbohydrate digestion inhibition effect was the highest. The extracts extracted at 20 ℃ showed 23.42%, 27.34% at 40 ℃, 41.75% at 80 ℃ and 34.32% at 100 ℃, respectively. (P <0.05), respectively.
실험예 3. 추출온도에 따른 히카마 물 추출물의 α-글루코시다아제 저해활성Experimental Example 3:? -Glucosidase inhibitory activity of Hikama water extract according to extraction temperature
추출온도에 따른 히카마 물 추출물의 α-글루코시다아제 저해활성을 보다 구체적으로 평가하기 위하여, 상기 실험예 2에서 추출온도를 55, 60, 65℃로 하여 추출한 것을 제외하고는 상기 실험예 2와 동일하게 실시하여 α-글루코시다아제 저해활성을 확인하였다. In order to more specifically evaluate the? -Glucosidase inhibitory activity of the Hikama water extract according to the extraction temperature, in Experimental Example 2, except that the extraction temperature was changed to 55, 60 and 65 ° C in Experimental Example 2 The same procedure was carried out to confirm the inhibitory activity of? -Glucosidase.
실험결과 도 3에 나타낸 바와 같이, 히카마 물 추출물의 제조 시 추출온도가 55℃인 경우 50.39%, 60℃인 경우 57.14%, 65℃인 겨우 52.34%로 우수한 탄수화물 분해효소 저해 활성을 나타났으며, 이같은 결과로부터 본 발명의 추출방법에 따라 추출용매로 물을 사용하고, 55~65℃의 추출온도에서 추출 시 탄수화물 분해효소 저해효과가 탁월한 히카마 추출물을 제조할 수 있음을 알 수 있었다.As shown in FIG. 3, the extract of Hikama water showed excellent carbohydrase inhibitory activity at 50.39% at 55 ° C, 57.14% at 60 ° C and 52.34% at 65 ° C . From these results, it was found that water extract was used as an extraction solvent according to the extraction method of the present invention, and Hycima extract having excellent carbohydrase inhibitory effect upon extraction at an extraction temperature of 55 to 65 ° C could be produced.
실험예 4. 히카마 추출물의 혈당강하효과Experimental Example 4 Effect of Hicoma Extract on Blood Glucose
상기 실시예 및 실험예들을 통해 탄수화물 소화효소 저해효과가 가장 높게 나타난 추출법인 추출용매로 물을 사용하고, 60℃의 추출온도에서 열수추출하여 제조한 히카마 추출물의 식후 혈당 강하 효과를 관찰하기 위하여 하기와 같은 실험하였다. To observe the hypoglycemic effect of Hikoma extract prepared by using water as the extraction solvent and extracting the hot water at the extraction temperature of 60 ° C, the carbohydrate digesting enzyme inhibition effect was highest through the above Examples and Experimental Examples The following experiment was conducted.
실험동물 및 당뇨유발Experimental animals and diabetes induction
동물실험을 위하여 4주령의 ICR mouse를 1주일간 일반식이로 적응시킨 후 실험에 사용하였다. ICR mouse에게 고형사료(Constant Nutrition, 미국)를 공급하여 평균체중이 35∼40g이 되게 사육한 다음, 12시간 공복상태의 ICR mouse에게 0.1M citrate buffer(pH 4.5)에 용해시킨 스트렙토조토신(STZ)을 복강에 주사하여 당뇨를 유발하였다. STZ 투여 1주일 후 공복상태의 ICR mouse 꼬리정맥으로부터 채혈하여 간이혈당계로 측정한 혈당이 250㎎/㎗ 이상일 때 당뇨병이 유발된 것으로 간주하여 실험에 사용하였다.Four weeks old ICR mice were adapted to the general diet for 1 week for the experiment. The ICR mice were fed with a solid diet (Constant Nutrition, USA), and the animals were weighed to an average weight of 35-40 g. ICR mice were fasted for 12 hours. Streptozotocin dissolved in 0.1 M citrate buffer (pH 4.5) ) Was injected into the abdominal cavity to induce diabetes. After 1 week of STZ administration, ICR mouse was used to test for diabetes mellitus when blood glucose level was 250mg / ㎗ or more.
경구 당부하 검사Oral glucose tolerance test
상기 준비된 실험동물을 스트렙토조토신으로 당뇨 유도한 당뇨생쥐와 정상생쥐를 대조군과 히카마 추출물 투여군, 아카보즈 투여군으로 나누었다. 대조군은 ICR mouse에 가용성 전분(2g/㎏, Sigma Co., USA)을, 아카보즈 투여군은 가용성 전분(2g/㎏) 및 아카보즈(100㎎/㎏)를, 히카마 추출물 투여군은 가용성 전분(2g/㎏) 및 히카마 추출물(200㎎/㎏)를 증류수에 용해하여 12시간 절식시킨 ICR mouse에 경구투여하였다. 투여 후 0, 30, 60, 120분 후에 꼬리정맥에서 채혈하여 혈당을 측정하였다. 각 시점의 혈당 증가치를 계산하여 혈당증가곡선을 구하고, 혈당증가곡선의 면적(area under the curve, AUC)을 계산하여 하기 도 4 및 5와 하기 표 1에 나타내었다. Diabetic mice and normal mice were divided into diabetic rats, streptozotocin-induced diabetic rats, control group, Hikama extract group, and acavos group. In the control group, soluble starch (2 g / kg, Sigma Co., USA) was administered to the ICR mouse, soluble starch (2 g / kg) and acarbose (100 mg / kg) 2 g / kg) and Hikoma extract (200 mg / kg) were dissolved in distilled water and orally administered to ICR mice fasted for 12 hours. Blood was collected from the tail vein at 0, 30, 60, and 120 minutes after the administration. The area under the curve (AUC) of the blood glucose increase curve was calculated by calculating the blood glucose increase value at each time point, and the area under the curve (AUC) was calculated.
도 4에 나타낸 바와 같이, 당뇨 생쥐의 경우 히카마 추출물 투여군이 투여 후 30, 60, 90, 120분에 혈당 증가가 각각 296.60, 342.00, 367.00, 329.67㎎/dl로 대조군(각각 296.00, 383.75, 402.50, 349.67㎎/dl)에 비해 낮게 나타남을 확인할 수 있었다. As shown in FIG. 4, in the case of diabetic mice, the increase in blood glucose was 296.60, 342.00, 367.00 and 329.67 mg / dl at 30, 60, 90 and 120 minutes after the administration of Hikoma extract, respectively (296.00, 383.75 and 402.50 , And 349.67 mg / dl), respectively.
또한 도 5에 나타낸 바와 같이, 정상 생쥐의 경우 히카마 추출물 투여군이 투여 후 30, 60, 90, 120분에 혈당 증가가 각각 88.16, 147.00, 139.83, 117.83㎎/dl, 대조군(각각 88.80, 175.33, 178.50, 151.68㎎/dl)에 비해 낮게 나타남을 확인할 수 있었다 .As shown in FIG. 5, in the case of the normal mice, the blood glucose levels were increased to 88.16, 147.00, 139.83 and 117.83 mg / dl at 30, 60, 90 and 120 minutes after administration of Hikoma extract, 178.50 and 151.68 mg / dl), respectively.
또한 히카마 추출물 투여군의 식후혈당증가 곡선의 면적(Area under the curve, AUC)은 상기 표 1에 나타낸 바와 같이, 히카마 추출물 투여군의 당뇨생쥐는 3537.9㎎·min/dl로 대조군(5814.9㎎·min/dl)에 비해 유의적으로 작게 나타났다. 따라서, 본 발명에 따른 히카마 추출물은 식후 혈당증가를 저해하는 효과가 있음을 확인할 수 있었다. As shown in Table 1, the area under the curve (AUC) of the postprandial blood glucose increase curve of the Hikoma extract administration group was 3537.9 mg · min / dl in the diabetic mice of the Hikama extract administration group and the control group (5814.9 mg · min / dl), respectively. Therefore, it was confirmed that the hycima extract according to the present invention has an effect of inhibiting an increase in postprandial blood glucose.
이상과 같은 결과로부터, 본 발명에 따라 제조한 히카마 추출물은 탄수화물 소화효소 저해효과가 탁월하며, 혈당강하에 현저한 탁월한 효과를 나타내어 당뇨병 치료제로 개발될 수 있을 뿐만 아니라, 당뇨병 관련 질환의 건강기능식품 조성물로 사용될 수 있을 것임을 예측할 수 있었다. From the above results, it can be concluded that the Hycima extract prepared according to the present invention is excellent in the inhibitory effect on carbohydrate digestion enzyme and exhibits a remarkable effect on blood glucose lowering, so that it can be developed as a therapeutic agent for diabetes, It could be predicted that it could be used as a composition.
제제예 1. 약학 제제의 제조Formulation Example 1. Preparation of pharmaceutical preparations
산제 제조 Acid production
상기 실시예 1에서 제조한 히카마 물 추출물 20㎎, 유당 100㎎ 및 탈트 10㎎을 혼합하고 기밀포에 충진하여 산제를 제조하였다.20 mg of Hikama water extract prepared in Example 1, 100 mg of lactose and 10 mg of talt were mixed and filled in airtight bags to prepare powders.
정제 제조Tablet manufacture
상기 실시예 1에서 제조한 히카마 물 추출물 10㎎, 옥수수전분 100㎎, 유당 100㎎ 및 스테아린산 마그네슘 2㎎을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.10 mg of Hikama water extract prepared in Example 1, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate were mixed and tableted according to a conventional preparation method.
캡슐제 제조Capsule preparation
통상의 캡슐제 제조방법에 따라 히카마 추출물 또는 분획물 10㎎, 결정성 셀룰로오스 3㎎, 락토오스 14.8㎎ 및 마그네슘 스테아레이트 0.2㎎을 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.10 mg of Hikoma extract or fraction, 3 mg of crystalline cellulose, 14.8 mg of lactose and 0.2 mg of magnesium stearate were mixed and filled in gelatin capsules according to a conventional capsule preparation method to prepare capsules.
주사제 제조Injection manufacturing
통상의 주사제의 제조방법에 따라 1앰플당(2mL) 상기 실시예 1에서 제조한 히카마 물 추출물 10㎎, 만니톨 180㎎, 주사용 멸균 증류수 2,974㎎ 및 Na2HPO4· 2H2O 26㎎으로 제조하였다.In accordance with the usual injection preparation method, 10 mg of Hikama water extract prepared in Example 1, 180 mg of mannitol, 2,974 mg of sterilized distilled water for injection, and 26 mg of Na 2 HPO 4 .2H 2 O were added per ampoule (2 mL) .
액제 제조Liquid preparation
통상의 액제의 제조방법에 따라 정제수에 상기 실시예 1에서 제조한 히카마 물 추출물 20㎎, 이성화당 10g 및 만니톨 5g을 가하여 용해시키고 레몬향을 적량 가한 다음 상기의 성분을 혼합하였다. 그 다음 정제수를 더 가하여 전체 100mL로 조절한 후 갈색병에 충진하고 멸균시켜 액제를 제조하였다.20 mg of Hikama water extract prepared in Example 1, 10 g of isomerized sugar, and 5 g of mannitol were dissolved in purified water, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed. Then, purified water was further added thereto, adjusted to a total volume of 100 mL, filled in a brown bottle, and sterilized to prepare a liquid preparation.
제제예 2. 식품 제제의 제조Formulation Example 2. Preparation of food preparation
건강식품 제조Health food manufacturing
상기 실시예 1에서 제조한 히카마 물 추출물 100㎎, 비타민 혼합물 적량, 비타민 A 아세테이트 70g, 비타민 E 1.0㎎, 비타민 B1 0.13㎎, 비타민 B2 0.15㎎, 비타민 B6 0.5㎎, 비타민 B12 0.2g, 비타민 C 10㎎, 비오틴 10g, 니코틴산아미드 1.7㎎, 엽산 50g, 판토텐산 칼슘 0.5㎎, 무기질 혼합물 적량, 황산제1철 1.75㎎, 산화아연 0.82㎎, 탄산마그네슘 25.3㎎, 제1인산칼륨 15㎎, 제2인산칼슘 55㎎, 구연산칼륨 90㎎, 탄산칼슘 100㎎ 및 염화마그네슘 24.8㎎을 혼합한 다음, 과립을 제조하고 통상의 방법에 따라 건강식품을 제조하였다. 이때, 상기 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하다.100 mg of Hikama water extract prepared in Example 1, a proper amount of vitamin A acetate, 70 g of vitamin A acetate, 1.0 mg of vitamin E, 0.13 mg of vitamin B1, 0.15 mg of vitamin B2, 0.5 mg of vitamin B6, 0.2 g of vitamin B12, 10 mg of biotin, 1.7 g of nicotinic acid amide, 50 g of folic acid, 0.5 mg of calcium pantothenate, 1.75 mg of ferrous sulfate, 0.82 mg of zinc oxide, 25.3 mg of magnesium carbonate, 15 mg of potassium phosphate, 55 mg of calcium, 90 mg of potassium citrate, 100 mg of calcium carbonate and 24.8 mg of magnesium chloride were mixed and granules were prepared and a health food was prepared according to a conventional method. At this time, although the composition ratio of the vitamin and mineral mixture is relatively mixed with the ingredient suitable for health food, it may be arbitrarily modified.
건강음료 제조Health drink manufacturing
통상의 건강음료 제조방법에 따라 상기 실시예 1에서 제조한 히카마 물 추출물 100㎎, 비타민 C 15g, 비타민 E(분말) 100g, 젖산철 19.75g, 산화아연 3.5g, 니코틴산아미드 3.5g, 비타민 A 0.2g, 비타민 B1 0.25g, 비타민 B2 0.3g 및 정량의 물을 혼합한 다음, 약 1시간 동안 85℃에서 교반 가열한 후 만들어진 용액을 여과하여 멸균된 2L 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관하여 건강음료를 제조하였다. 이때, 상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만 수요계층이나, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.100 mg of Hikama water extract, 15 g of vitamin C, 100 g of vitamin E (powder), 19.75 g of iron lactate, 3.5 g of zinc oxide, 3.5 g of nicotinic acid amide, 3.5 g of vitamin A 0.25 g of vitamin B1, 0.3 g of vitamin B2 and a predetermined amount of water were mixed and heated at 85 DEG C for about 1 hour with stirring. The resulting solution was filtered and sterilized in a sterilized 2 L vessel, To produce a health drink. At this time, although the composition ratio of the ingredients suitable for the beverage is comparatively mixed, the mixture ratio may be arbitrarily varied according to the demand, the demanded country, the intended use, and the regional or national preference.
비록 본 발명이 상기에 언급된 바람직한 실시예로서 설명되었으나, 발명의 요지와 범위로부터 벗어남이 없이 다양한 수정이나 변형을 하는 것이 가능하다. 또한 첨부된 청구 범위는 본 발명의 요지에 속하는 이러한 수정이나 변형을 포함한다.Although the present invention has been described in terms of the preferred embodiments mentioned above, it is possible to make various modifications and variations without departing from the spirit and scope of the invention. It is also to be understood that the appended claims are intended to cover such modifications and changes as fall within the scope of the invention.
Claims (10)
상기 건조한 히카마를 분쇄하여 분말화하는 단계;
상기 히카마 분말에 추출용매를 가하여 초음파 분쇄하면서 50~80℃에서 4~6시간 동안 추출하는 단계; 및
상기 히카마 추출물을 감압농축하는 단계;
를 포함하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법.Washing and drying the hicamas;
Pulverizing and drying the dried hicamas;
Adding an extraction solvent to the Hikama powder and extracting it at 50 to 80 ° C for 4 to 6 hours while ultrasonically pulverizing it; And
Concentrating the Hicoma extract at reduced pressure;
Wherein the extract has a carbohydrate digestive enzyme inhibitory effect.
상기 추출용매는 물, 50% 에탄올 및 50% 메탄올 중 선택된 어느 하나 이상인 것을 특징으로 하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법.The method according to claim 1,
Wherein the extraction solvent is at least one selected from water, 50% ethanol and 50% methanol.
상기 추출용매는 물인 것을 특징으로 하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법.The method according to claim 1,
Wherein the extraction solvent is water. ≪ RTI ID = 0.0 > 11. < / RTI >
상기 초음파 분쇄는 초음파 분쇄기를 이용하여 10~60초 동안 작동시킨 후 10~20초 동안 중지시키는 동작을 반복하여 수행되는 것을 특징으로 하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법.The method according to claim 1,
Wherein the ultrasonic pulverization is performed by using an ultrasonic grinder for 10 to 60 seconds and then for 10 to 20 seconds to be repeatedly performed.
상기 추출은 초음파 분쇄기를 이용하여 55~65℃에서 5시간 동안 추출하는 과정을 3회 반복하여 수행되는 것을 특징으로 하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법.The method according to claim 1,
Wherein the extraction is carried out by repeating the process of extracting at 55 to 65 ° C for 5 hours using an ultrasonic grinder three times.
상기 히카마 추출물은 α-글루코시다아제 저해활성을 갖는 것을 특징으로 하는 탄수화물 소화효소 저해 효과를 가지는 히카마 추출물의 제조방법.The method according to claim 1,
Wherein the Hicoma extract has an inhibitory activity against alpha-glucosidase.
상기 히카마 추출물은 추출용매로 물을 사용하여 55~65℃의 추출온도에서 열수추출한 히카마 물 추출물인 것을 특징으로 하는 당뇨병 관련 질환의 예방 또는 치료용 약학 조성물.8. The method of claim 7,
The pharmaceutical composition for prevention or treatment of diabetes related diseases, wherein the Hicoma extract is Hikoma water extract obtained by hot water extraction at an extraction temperature of 55 to 65 占 폚 using water as an extraction solvent.
상기 히카마 추출물은 조성물 총 중량에 대하여 0.01~95중량%로 포함되는 것을 특징으로 하는 당뇨병 관련 질환의 예방 또는 치료용 약학 조성물.8. The method of claim 7,
The pharmaceutical composition for preventing or treating a diabetes related disease according to claim 1, wherein the hiccama extract is contained in an amount of 0.01 to 95% by weight based on the total weight of the composition.
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