KR102293400B1 - Composition for eradicating Korean Helicobacter pylori and its use - Google Patents
Composition for eradicating Korean Helicobacter pylori and its use Download PDFInfo
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- KR102293400B1 KR102293400B1 KR1020200029343A KR20200029343A KR102293400B1 KR 102293400 B1 KR102293400 B1 KR 102293400B1 KR 1020200029343 A KR1020200029343 A KR 1020200029343A KR 20200029343 A KR20200029343 A KR 20200029343A KR 102293400 B1 KR102293400 B1 KR 102293400B1
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Abstract
Description
본 발명은 한국형 헬리코박터 파일로리 제균용 조성물에 관한 것으로, 더 상세하게는 꾸지뽕나무 열매 분획물을 유효성분으로 포함하는 한국형 헬리코박터 파일로리 제균용 조성물 및 이의 용도에 관한 것이다.The present invention relates to a Korean-type Helicobacter pylori sterilizing composition, and more particularly, to a Korean-type Helicobacter pylori sterilizing composition and use thereof, comprising a Cuji mulberry fruit fraction as an active ingredient.
헬리코박터 파일로리는 위에 기생하는 나선형 모양의 세균으로 인체에 만성, 급성위염, 위궤양 및 위암 등을 유발하는 것으로 알려진 병원체이다. 한국인의 60% 이상이 헬리코박터 파일로리에 감염된 것으로 알려져 있다. 염증이 위장내 점막하층까지 퍼지게 되면 위궤양으로 발전하게 되고, 위암으로 바로 진행되는 것은 아니지만, 헬리코박터 파일로리에 감염되면 위암으로 진행될 가능성이 3~5배 정도 높아지는 것으로 알려져 있다. Helicobacter pylori is a spiral-shaped bacterium that parasitizes in the stomach and is a pathogen known to cause chronic, acute gastritis, gastric ulcer and gastric cancer in the human body. It is known that more than 60% of Koreans are infected with Helicobacter pylori. When inflammation spreads to the submucosal layer of the stomach, it develops into a gastric ulcer, and does not progress directly to gastric cancer.
헬리코박터 파일로리의 제균을 위해서 테트라사이클린(tetracycline)이나 아목시실린(amoxicillin)과 같은 항생제가 병용 사용되었고, 최근에는 3중 치료법으로 2~3주 동안 치료를 요구하고 있다. 미국 국립보건원에 따르면, 위궤양 및 소화성 질환 치료를 목적으로 헬리코박터 파일로리를 타겟으로 하는 항생제의 사용을 허가하였지만, 헬리코박터를 비롯한 그램 음성균 전체를 타겟으로 하는 항생제의 장기적 복용은 유해균뿐 아니라 유익균의 공동 사멸, 항생제 내성균의 출현, 장내미생물균총 교란 등 다양한 부작용을 유도하여 궁극적인 치료법이 되지 못하는 것이 실정이다. For the eradication of Helicobacter pylori, antibiotics such as tetracycline or amoxicillin were used in combination, and recently, a triple therapy is required for 2-3 weeks. According to the US National Institutes of Health, the use of antibiotics targeting Helicobacter pylori has been approved for the purpose of treating gastric ulcer and peptic diseases. The reality is that it is not the ultimate treatment because it induces various side effects such as the emergence of antibiotic-resistant bacteria and disturbance of the intestinal microflora.
한편 한국인의 위에서 헬리코박터 파일로리 51(Helicobacter pylori 51)가 분리되었다. 이 한국형 헬리코박터 파일로리 균주는 기존에 밝혀진 서양인의 위에서 분리된 균주 헬리코박터 파일로리 26695와 헬리코박터 파일로리 J99에서 나타나지 않는 특이적으로 유전자들을 포함하고 있다. Meanwhile, Helicobacter pylori 51 was isolated from a Korean stomach. This Korean-type Helicobacter pylori strain contains specific genes that do not appear in Helicobacter pylori 26695 and Helicobacter pylori J99 isolated from the stomach of Westerners previously identified.
따라서 항생제의 사용에 따른 내성균 출현 등의 부작용은 적으면서도 한국형 헬리코박터 파일로리를 효과적으로 제균할 수 있는 천연물 유래의 대체 약제의 개발이 요구되고 있다.Therefore, there is a need to develop an alternative drug derived from a natural product that can effectively sterilize Korean Helicobacter pylori while having few side effects such as emergence of resistant bacteria due to the use of antibiotics.
이에 본 발명자들은 종래 기술에서의 요구에 부응하기 위해 지속적으로 연구한 결과, 꾸지뽕나무 열매 분획물이 놀랍게도 힌국형 헬리코박터 파일로리 제균 효과가 우수한 것을 확인하여 본 발명을 완성하게 되었다. Accordingly, as a result of continuous research to meet the needs of the prior art, the present inventors have completed the present invention by confirming that the Cuji mulberry fruit fraction has surprisingly excellent antibacterial effect on H. pylori Helicobacter pylori.
따라서 본 발명의 목적은 꾸지뽕나무 열매 분획물을 유효성분으로 포함하는 한국형 헬리코박터 파일로리 제균용 조성물을 제공하는 것이다.Accordingly, it is an object of the present invention to provide a composition for sterilization of Helicobacter pylori in Korea, which contains a fraction from the fruit of the mulberry tree as an active ingredient.
본 발명의 또 다른 목적은 꾸지뽕나무 열매 분획물을 포함하는 한국형 헬리코박터 파일로리 치료용 약제학적 조성물을 제공하는 것이다. Another object of the present invention is to provide a pharmaceutical composition for the treatment of Korean Helicobacter pylori containing a fraction from the fruit of the mulberry tree.
본 발명의 또 다른 목적은 꾸지뽕나무 열매 분획물을 포함하는 한국형 헬리코박터 파일로리 개선용 건강기능성 식품을 제공하는 것이다. Another object of the present invention is to provide a health functional food for improving Korean-type Helicobacter pylori containing a Curcuma mulberry fruit fraction.
상기 본 발명의 목적을 달성하기 위하여, 본 발명은 꾸지뽕나무 열매 분획물을 유효성분으로 포함하는 한국형 헬리코박터 파일로리 제균용 조성물을 제공한다. In order to achieve the above object of the present invention, the present invention provides a composition for sterilization of Helicobacter pylori in Korea comprising a fraction from the fruit of the mulberry tree as an active ingredient.
꾸지뽕나무 (Cudrania tricuspidata B.)는 뽕나무과에 속하는 낙엽성 소교목 또는 관목으로서 우리나라, 중국, 일본 등지에 자생하는 식물이다. 꾸지뽕나무 잎은 한방에서 습진, 유행성 이하선염, 당뇨, 만성요퇴통, 타박상 등의 치료에 사용되며, 줄기는 주로 부인들의 붕중과 혈결의 치료에, 열매는 청열과 양혈을 다스리는 데 이용되고 있는 것으로 알려져 있다. 또한 민간에서 열매는 강장, 중풍, 이뇨 및 진해 등의 치료약으로 이용되어 왔다. 그러나, 꾸지뽕나무 열매의 한국형 헬리코박터 파일로리 제균 효과는 알려진 바 없다. Cudrania tricuspidata B. ) is a deciduous small tree or shrub belonging to the Morus family, and is a plant native to Korea, China, and Japan. It is known that mulberry leaves are used in oriental medicine to treat eczema, mumps, diabetes, chronic low back pain, bruises, etc. have. In addition, in folklore, the fruit has been used as a remedy for tonicity, paralysis, diuresis, and cough. However, the bactericidal effect of Helicobacter pylori in Korea of Cuji mulberry fruit is not known.
본 발명에서, 꾸지뽕나무 열매 분획물은 꾸지뽕나무 열매의 용매 추출물을 추가적으로 분획(fractionation)한 분획 추출물을 의미한다. In the present invention, the Cuji mulberry fruit fraction refers to a fractional extract obtained by additionally fractionating the solvent extract of Cuji mulberry fruit.
본 발명에서, 꾸지뽕나무 열매 분획물은 꾸지뽕나무 열매를 용매를 사용하여 추출한 추출물을 추가로 비극성 용매로 분획한 분획물을 사용할 수 있다. 바람직하게는 꾸지뽕나무 열매 분획물은 꾸지뽕나무 열매를 메탄올로 추출한 추출물의 클로로포름 또는 헥산 분획물이다. In the present invention, the Cuji mulberry fruit fraction can be used as a fraction obtained by further fractionating the extract obtained by extracting the Cuji mulberry fruit using a solvent with a non-polar solvent. Preferably, the Cudrania fruit fraction is a chloroform or hexane fraction of an extract obtained by extracting the Cucurbita fruit with methanol.
본 발명에서 꾸지뽕나무 열매 분획물은 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수도 있다.In the present invention, the mulberry fruit fraction may be prepared in a powder state by an additional process such as freeze-drying or spray-drying.
본 발명에 따른 조성물의 유효성분인 꾸지뽕나무 열매의 헥산 분획물 또는 클로로포름 분획물은 한국형 헬리코박터 파일로리 제균(생육저해)에 탁월한 효과를 나타낸다 (도 2a, 도 2 b). 구체적으로는 꾸지뽕나무 열매의 헥산 분획물 또는 클로로포름 분획물은 메트로니다졸 (알려진 헬리코박터 파일로리 제균제) 보다 더 우수한 한국형 헬리코박터 파일로리 생육저해 효능을 나타냈으며, 특히 꾸지뽕나무 열매의 클로로포름 분획물은 메트로니다졸에 비하여 1.5배 증진된 한국형 헬리코박터 파일로리 제균 효능을 나타낸다 (도 2b).The active ingredient of the composition according to the present invention, the hexane fraction or the chloroform fraction of the mulberry tree fruit, exhibits an excellent effect on sterilization (inhibition of growth) of Korean Helicobacter pylori ( FIGS. 2a and 2b ). Specifically, the hexane fraction or chloroform fraction of Cucurbita fruit showed a better Korean Helicobacter pylori growth inhibitory effect than metronidazole (a known Helicobacter pylori disinfectant). It shows the efficacy of Helicobacter pylori eradication (Fig. 2b).
본 발명에 따른 꾸지뽕나무 열매 분획물은 한국형 헬리코박터 파일로리의 제균 효과를 나타내는 농도 범위에서는 세포 독성이 없다 (도 3). The Cuji mulberry fruit fraction according to the present invention has no cytotoxicity in the concentration range showing the bactericidal effect of Korean Helicobacter pylori (FIG. 3).
본 발명의 꾸지뽕나무 열매 분획물은 한국형 헬리코박터 파일로리 치료용 약제학적 조성물로 제형될 수 있다. The fruit fraction of Cudrania biloba of the present invention may be formulated as a pharmaceutical composition for the treatment of Korean Helicobacter pylori.
본 발명에 따른 약제학적 조성물은 꾸지뽕나무 열매의 헥산 분획물 또는 클로로포름 분획물의 약제학적 유효량 및 약제학적으로 허용되는 담체를 포함할 수 있다. The pharmaceutical composition according to the present invention may contain a pharmaceutically effective amount of a hexane fraction or a chloroform fraction of Cucurbita fruit and a pharmaceutically acceptable carrier.
본 발명에서 ‘약제학적 유효량’은 꾸지뽕나무 열매의 헥산 분획물 또는 클로로포름 분획물의 효능 또는 활성을 달성하는 데 충분한 양을 의미한다. In the present invention, the 'pharmaceutically effective amount' means an amount sufficient to achieve the efficacy or activity of the hexane fraction or chloroform fraction of Cudrania fruit.
본 발명에서 ‘약제학적으로 허용되는 담체’는 제형시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산 칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및/또는 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약제학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다. In the present invention, the 'pharmaceutically acceptable carrier' is commonly used in formulation, and includes lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, fine crystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and/or mineral oil, and the like. no. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like, in addition to the above components.
본 발명에 따른 약제학적 조성물은 경구 또는 비경구 투여할 수 있으며, 바람직하게는 경구 투여 방식으로 적용된다. The pharmaceutical composition according to the present invention may be administered orally or parenterally, and is preferably applied by oral administration.
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하게 처방될 수 있다. 본 발명의 약제학적 조성물의 일반적인 투여량은 성인 기준으로 0.001 ~ 100 ㎎/kg 범위 내이다.A suitable dosage of the pharmaceutical composition of the present invention is variously prescribed depending on factors such as formulation method, administration mode, age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and reaction sensitivity of the patient. can be A typical dosage of the pharmaceutical composition of the present invention is in the range of 0.001 to 100 mg/kg based on an adult.
본 발명에 따른 꾸지뽕나무 열매의 분획물은 한국형 헬리코박터 파일로리 개선용 건강기능성 식품으로 제공될 수 있다. The fraction of Cuji mulberry fruit according to the present invention may be provided as a health functional food for improving Korean Helicobacter pylori.
본 발명의 건강기능성 식품은 꾸지뽕나무 열매의 헥산 분획물 또는 클로로포름 분획물을 유효성분으로 포함한다.The health functional food of the present invention comprises a hexane fraction or a chloroform fraction of Cuji mulberry fruit as an active ingredient.
본 발명의 건강기능성 식품은 유효성분으로서 꾸지뽕나무 열매 분획물에 추가로 식품 제조시에 통상적으로 첨가가되는 첨가제를 포함할 수 있다. 첨가제로는 예를 들어, 단백질, 당류, 지방, 영양소, 조미제 및 향미제가 있다. 당류의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이 사용될 수 있다. 향미제로서 천연 향미제 [타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다.The health functional food of the present invention may include additives that are normally added during food production in addition to the Cuji mulberry fruit fraction as an active ingredient. Additives include, for example, proteins, sugars, fats, nutrients, seasonings and flavoring agents. Examples of saccharides include monosaccharides such as glucose, fructose, and the like; disaccharides such as maltose, sucrose, oligosaccharides and the like; and polysaccharides, for example, conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents, natural flavoring agents [taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.)) and synthetic flavoring agents (saccharin, aspartame, etc.) can be used.
본 발명에서 건강기능성 식품은 정제, 환제, 분말, 액상차, 음료, 과자, 발효식품, 통조림, 우유가공식품, 육류가공식품, 식품첨가제 등의 형태일 수 있으나, 이에 한정되지는 않는다.In the present invention, the health functional food may be in the form of tablets, pills, powder, liquid tea, beverages, confectionery, fermented food, canned food, processed milk food, processed meat food, food additives, and the like, but is not limited thereto.
본 발명에 따른 꾸지뽕나무 열매 분획물을 유효성분으로 포함하는 조성물 및 약제학적 조성물은, 종래 알려진 헬리코박터 파일로리 제균제에 필적하거나 훨씬 증진된 한국형 헬리코박터 파일로리 생육저해 효능을 가지면서도 독성도 없어서, 한국형 헬리코박터 파일로리 제균 및 한국형 헬리코박터 파일로리로 인해 유발된 위장 질환의 치료에 매우 유용하다.The composition and pharmaceutical composition comprising the Cuji mulberry fruit fraction according to the present invention as an active ingredient have a Korean-type Helicobacter pylori growth inhibitory effect that is comparable to or significantly improved to a conventional Helicobacter pylori disinfectant, but there is no toxicity, so Korean Helicobacter pylori eradication and gastrointestinal diseases caused by Korean Helicobacter pylori.
도 1은 본 발명에 따른 꾸지뽕나무 열매 분획물의 제조 공정도의 일례이다.
도 2a 및 도 2b는 본 발명에 따른 꾸지뽕나무 열매 분획물의 헬리코박터 파일로리 제균 효능을 분석한 결과 그래프이다.
도 3은 본 발명에 따른 꾸지뽕나무 열매 분획물의 세포독성을 분석한 결과 그래프이다. 1 is an example of a manufacturing process diagram of a mulberry fruit fraction according to the present invention.
2a and 2b are graphs of the results of analyzing the Helicobacter pylori eradication efficacy of the Cuji mulberry fruit fraction according to the present invention.
3 is a graph showing the result of analyzing the cytotoxicity of the Cuji mulberry fruit fraction according to the present invention.
이하, 본 발명의 이해를 돕기 위하여 구체적인 실시예를 통하여 본 발명의 구성 및 효과를 보다 상세히 설명하기로 한다. 그러나 하기 실시예는 본 발명을 보다 명확하게 이해시키기 위하여 예시한 것일 뿐이며, 본 발명의 권리범위가 하기 실시예에 의해 한정되는 것은 아니다. Hereinafter, the configuration and effects of the present invention will be described in more detail through specific examples to help the understanding of the present invention. However, the following examples are merely illustrative in order to understand the present invention more clearly, and the scope of the present invention is not limited by the following examples.
제조예 1: 꾸지뽕나무 열매 분획물 제조Preparation Example 1: Preparation of Cuji mulberry fruit fraction
산청 및 진주에서 재배되고 있는 꾸지뽕나무 열매를 수확하고 건조하여 7 kg의 건조중량을 얻었고 이 꾸지뽕나무 열매를 90% 메탄올 20 L를 이용하여 침지 추출을 3반복 실시한 후, 추출된 용액(추출액)을 No.2 종이필터를 이용하여 여과하였으며 대형회전감압농축를 이용하여 농축하여 약 1.09 kg의 조추출물을 얻었다. After harvesting and drying Cuji mulberry fruits cultivated in Sancheong and Jinju, a dry weight of 7 kg was obtained. It was filtered using a No.2 paper filter and concentrated using a large rotary vacuum concentration to obtain about 1.09 kg of a crude extract.
상기 조추출물 1.09kg을 5% 메탄올 1 L에 현탁시킨 후 헥산, 크로로포름, 에틸아세테이트, 물(H2O) 층으로 용매 분획을 반복 시행하여 분획물들을 모은 후 회전감압농축기를 이용하였다 (도 1). After suspending 1.09 kg of the crude extract in 1 L of 5% methanol , solvent fractionation was repeatedly performed with hexane, chloroform, ethyl acetate, and water (H 2 O) layers to collect fractions, and then a rotary vacuum concentrator was used (Fig. One).
헥산 분획물은 4.0%, 클로로포름 분획물은 3.5% 및 에틸아세테이트 분획물은 2.5%, 물 분획물은 90.0%의 수율을 나타내었다. The hexane fraction was 4.0%, the chloroform fraction was 3.5%, the ethyl acetate fraction was 2.5%, and the water fraction showed a yield of 90.0%.
시험예 1: 한국형 헬리코박터 파일로리 제균 효능 분석Test Example 1: Analysis of Efficacy of Sterilization of Korean Helicobacter pylori
제조예 1에서 제조된 각 꾸지뽕나무 열매 분획물의 한국형 헬리코박터 파일로리 제균 효능을 분석하였다. The bactericidal efficacy of Korean Helicobacter pylori was analyzed for each fraction of Cuji mulberry fruit prepared in Preparation Example 1.
한국형 헬리코박터 파일로리 균주로는 Helicobacter pylori strain 51 (헬리코박터 파일로리 분리균주은행(HKTCC), 경상대학교 소재)을 이용하여 측정하였다. As a Korean Helicobacter pylori strain, Helicobacter pylori strain 51 (Helicobacter pylori isolate bank (HKTCC), located in Gyeongsang National University) was used.
상기 균주를 말 혈청(Gibco, New York, USA)이 10% 첨가된 브루셀라 한천배지(BD company, Sparks, MD, USA) 위에 떨어뜨린 후 일회용 플라스틱 접종자를 이용하여 표면에 고르게 도말한 후 37℃, 습도 100%, 10% CO2 배양기에서 2 ~ 3일간 배양하였으며, 24 시간마다 계대배양하였다. 브루셀라 한천배지에 배양한 균을 접종자로 긁은 다음 브루셀라 액체배지에 현탁하였다. 6-웰 플레이트에서 현탁한 균액의 광학밀도(OD 600)를 측정한 다음 그 값이 0.2가 되도록 희석하였다. 희석한 균액을 2% 함유한 브루셀라 한천배지를 준비하였다. After dropping the strain on Brucella agar medium (BD company, Sparks, MD, USA) to which 10% of horse serum (Gibco, New York, USA) was added, spread evenly on the surface using a disposable plastic inoculator, and then at 37°C,
제조예 1에서 제조된 각 꾸지뽕나무 열매 분획물을 최종농도 10 mg/ml 샘플을 500 ㎍/ml, 250 ㎍/ml, 125 ㎍/ml, 62 ㎍/ml, 및 25 ㎍/ml 농도로 처리하여 0.45 ㎛ 시린지 필터(Whatman, USA)로 여과하여 8 mm 종이디스크(Advantec, Japan)에 30 ㎕ 씩 떨어뜨려 흡수시켰다. 각각의 종이디스크를 상기에 준비된 한천배지에 올려놓고 1시간 정도 실온에서 확산시킨 후, CO2 배양기에서 2일간 배양하면서 종이디스크 주위에 생성된 클리어 존(clear zone)의 지름을 측정하였다. Each fraction prepared in Preparation Example 1 was treated with a final concentration of 10 mg/ml sample at a concentration of 500 μg/ml, 250 μg/ml, 125 μg/ml, 62 μg/ml, and 25 μg/ml to 0.45 It was filtered with a ㎛ syringe filter (Whatman, USA) and was absorbed by dropping 30 μl each on an 8 mm paper disk (Advantec, Japan). Each of the paper disks was placed on the agar medium prepared above and spread at room temperature for about 1 hour, and then incubated in a CO 2 incubator for 2 days while measuring the diameter of a clear zone created around the paper disk.
양성대조군으로써 기존에 판매되고 있는 제균제인 메트로니다졸 (Metronidazole)를 사용하여 동일한 방식으로 측정하였다.As a positive control, metronidazole, an existing disinfectant, was used as a positive control and measured in the same manner.
MIC의 경우 최소억제농도로서 10% 정도의 억제효과가 보이는 농도를 기재하였고 MIC50의 경우 50% 이상의 억제효과가 나타나는 농도를 환산하였다. In the case of MIC, the concentration showing the inhibitory effect of about 10% was described as the minimum inhibitory concentration, and in the case of MIC 50, the concentration showing the inhibitory effect of 50% or more was converted.
그 결과를 표 1 및 도 2a, 도 2b에 나타내었다. The results are shown in Table 1 and FIGS. 2A and 2B.
추출물methanol
extract
분획물 chloroform
fraction
표 1 및 도 2a, 도 2b에 도시된 바와 같이, 꾸지뽕나무 열매의 헥산 분획물 및 클로로포름 분획물은 양성대조군인 메트로니다졸 보다 우수한 한국형 헬리코박터 파일로리 제균 효과를 나타내었고, 특히 꾸지뽕나무 열매의 클로로포름 분획물에서는 양성대조군 보다 약 1.5배 증진된 한국형 헬리코박터 파일로리의 제균 효과를 나타냈다. As shown in Table 1 and FIGS. 2a and 2b, the hexane fraction and chloroform fraction of Cudrania fruit showed a superior Korean Helicobacter pylori eradication effect than metronidazole, which is a positive control group. It showed the antibacterial effect of Korean Helicobacter pylori, which was improved by about 1.5 times.
꾸지뽕나무 열매의 메탄올 추출물과 꾸지뽕나무 열매의 에틸아세테이트 분획물 및 물 분획물은 한국형 헬리코박터 파일로리의 제균 효과를 보이지 않았다. Methanol extract of Cucurbita fruit, ethyl acetate fraction and water fraction of Cudrania fruit did not show bactericidal effect on Korean Helicobacter pylori.
시험예 2: 세포독성 분석Test Example 2: Cytotoxicity analysis
제조예 1에서 제조된 각 꾸지뽕나무 열매 분획물과 꾸지뽕 메탄올(70%) 추출물의 세포독성을 분석하였다. The cytotoxicity of each Cuji mulberry fruit fraction and methanol (70%) Cuji mulberry extract prepared in Preparation Example 1 was analyzed.
구체적으로 마우스대식세포(Raw 264.7 세포, KCLB 40071)를 페니실린과 스트렙토마이신이 각각 100 unit/ml 첨가되고 10% FBS가 함유된 DMEM 배지에서 5%의 CO2하에 37℃로 하였으며 2일 간격으로 계대배양하였다. Raw 264.7 세포의 농도를 2x105 cells/ml으로 조정하여 96 웰 플레이트에 분주하고 각각의 분획물 및 메탄올 추출물을 250 ㎍/ml으로 처리한 다음 24시간을 배양시킨 후에 MTT 분석시약 (CellTiter 96® aqueousnon-radioactive cell proliferation assay reagent; Promega,WI, USA)을 첨가하여 배양기(37℃, 5% CO2)에서 4시간 반응시킨 후 멀티마이크로플레이트 리더기(Multimicroplate reader-SpectraMax M5; Molecular Devices, USA)로 450 nm에서 흡광도를 측정하여 대조군의 흡광도와 비교하여 세포생존율을 백분율로(%)로 환산하여, 세포 독성 여부를 확인하였고, 그 결과를 도 3에 나타냈다. Specifically, mouse macrophages (Raw 264.7 cells, KCLB 40071) were each added to 100 unit/ml penicillin and streptomycin and 5% CO 2 in DMEM medium containing 10% FBS at 37° C. and passaged every 2 days. cultured. The concentration of Raw 264.7 cells was adjusted to 2x10 5 cells/ml and dispensed in a 96-well plate, each fraction and methanol extract were treated with 250 μg/ml, and then incubated for 24 hours with MTT assay reagent (CellTiter 96® aqueous non- Radioactive cell proliferation assay reagent; Promega, WI, USA) was added and reacted for 4 hours in an incubator (37° C., 5% CO 2 ), followed by a multimicroplate reader (Multimicroplate reader-SpectraMax M5; Molecular Devices, USA) at 450 nm. By measuring the absorbance and converting the cell viability into a percentage (%) compared with the absorbance of the control, cytotoxicity was confirmed, and the result is shown in FIG. 3 .
도 3에 도시된 바와 같이, 본 발명에 따른 꾸지뽕나무 열매 분획물은 모두 독성이 없는 것으로 확인되었다. As shown in FIG. 3 , it was confirmed that all of the Cuji mulberry fruit fractions according to the present invention were non-toxic.
Claims (6)
상기 분획물은 꾸지뽕나무 열매의 클로로포름 분획물 또는 헥산 분획물이고,
상기 한국형 헬리코박터 파일로리는 헬리코박터 파일로리 51인 것인 조성물.
A composition for sterilization of Helicobacter pylori in Korea, comprising a fraction from the mulberry tree fruit as an active ingredient,
The fraction is a chloroform fraction or a hexane fraction of Cuji mulberry fruit,
The Korean-type Helicobacter pylori composition is Helicobacter pylori 51.
상기 분획물은 꾸지뽕나무 열매의 클로로포름 분획물 또는 헥산 분획물이고,
상기 한국형 헬리코박터 파일로리는 헬리코박터 파일로리 51인 것인 조성물.
A pharmaceutical composition for the treatment of Helicobacter pylori infection in Korea, comprising a pharmaceutically effective amount of the Cuji mulberry fruit fraction and a pharmaceutically acceptable carrier,
The fraction is a chloroform fraction or a hexane fraction of Cuji mulberry fruit,
The Korean-type Helicobacter pylori composition is Helicobacter pylori 51.
상기 분획물은 꾸지뽕나무 열매의 클로로포름 분획물 또는 헥산 분획물이고,
상기 한국형 헬리코박터 파일로리는 헬리코박터 파일로리 51인 것인 건강기능성 식품.As a health functional food for improving Korean Helicobacter pylori infection,
The fraction is a chloroform fraction or a hexane fraction of Cuji mulberry fruit,
The Korean-type Helicobacter pylori is a health functional food that is Helicobacter pylori 51.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20040072913A (en) | 2003-02-11 | 2004-08-19 | 한국생명공학연구원 | Novel genes of Helicobacter pylori 51 strain isolated from a Korean patient |
| KR20190132891A (en) | 2018-05-21 | 2019-11-29 | (주) 피러스 | composition for removing helicobacter pyloricomprising extract from pine-tree by-product |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20040072913A (en) | 2003-02-11 | 2004-08-19 | 한국생명공학연구원 | Novel genes of Helicobacter pylori 51 strain isolated from a Korean patient |
| KR20190132891A (en) | 2018-05-21 | 2019-11-29 | (주) 피러스 | composition for removing helicobacter pyloricomprising extract from pine-tree by-product |
Non-Patent Citations (3)
| Title |
|---|
| Prev. Nutr. Food Sci. 2019* * |
| 여희동, 경상대학교 대학원 석사학위 논문 (2009) 1부.* * |
| 제31회 한국생명과학회 학술발표회 논문초록, P41 (2001) 1부.* * |
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