KR20150092044A - A composition for treating or preventing tuberculosis comprising Conjugated(10E,12Z)-Linoleic acid - Google Patents
A composition for treating or preventing tuberculosis comprising Conjugated(10E,12Z)-Linoleic acid Download PDFInfo
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- KR20150092044A KR20150092044A KR1020150096987A KR20150096987A KR20150092044A KR 20150092044 A KR20150092044 A KR 20150092044A KR 1020150096987 A KR1020150096987 A KR 1020150096987A KR 20150096987 A KR20150096987 A KR 20150096987A KR 20150092044 A KR20150092044 A KR 20150092044A
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- tuberculosis
- composition
- linoleic acid
- present
- preventing
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Abstract
Description
본 발명은 10E,12Z 공액리놀레산(Conjugated(10E,12Z)-Linoleic acid)을 유효성분으로 포함하는 결핵의 치료, 개선 또는 예방용 약학 조성물, 예방 또는 개선용 건강식품 조성물, 예방 또는 개선용 사료 조성물, 치료, 개선 또는 예방용 동물의약품 조성물, 예방 또는 개선용 의약외품 조성물에 관한 것이다. 보다 구체적으로, 10E,12Z 공액리놀레산(Conjugated(10E,12Z)-Linoleic acid)은 다양한 종류의 동식물에 함유되어 있는 천연물 유래 활성 지방산 및 생리활성을 나타내는 물질로서 단일물질인 10E,12Z 공액리놀레산(Conjugated(10E,12Z)-Linoleic acid)은 결핵균(mycobacterium tuberculosis)의 분열증식 및 성장을 억제하는 효과가 있으므로 결핵의 치료, 개선 또는 예방용 약학 조성물, 예방 또는 개선용 건강식품 조성물, 예방 또는 개선용 사료 조성물, 치료, 개선 또는 예방용 동물의약품 조성물 및 예방 또는 개선용 의약외품 조성물로 유용하다.
The present invention relates to a pharmaceutical composition for the treatment, improvement or prevention of tuberculosis containing 10E, 12Z conjugated linoleic acid (Conjugated (10E, 12Z) -Linoleic acid) as an active ingredient, a health food composition for prevention or improvement, , An animal pharmaceutical composition for the treatment, improvement or prevention, and a quasi-product composition for prevention or improvement. More specifically, 10E, 12Z conjugated linoleic acid (Conjugated (10E, 12Z) -Linoleic acid) is a natural substance-derived active fatty acid and a substance exhibiting physiological activity contained in various kinds of animals and plants, and is a
수백 년 전만 해도 천연두, 홍역, 콜레라, 독감바이러스, 폐결핵, 흑사병과 같은 전염성 질병은 어린이와 성인들을 포함하는 전 인류에 급격한 사망을 초래하는 고위험 전염성 질병이었다. 이러한 전염성 질병은 수세기에 걸쳐 수천만 명의 사람을 사망시켰으며 현재까지도 일부 전염성 질병의 억제는 어려움에 처해있는 실정이다. 이러한 전염성 질병의 억제를 위한 연구 중 페니실린의 발견은 항생물질에 의한 병원성 미생물의 억제 또는 치료, 질병억제 및 건강보건향상과 수명연장에 큰 변화를 유발시키는 전환점이 되었다. 그렇지만, 아직도 병원성 미생물에 의한 직접 혹은 간접적인 피해는 경제, 환경, 의학 및 보건학적으로 다양한 문제와 질병을 유발시키고 있다. Hundreds of years ago, infectious diseases such as smallpox, measles, cholera, flu virus, pulmonary tuberculosis and plague were high-risk infectious diseases that caused rapid deaths in all human beings, including children and adults. These infectious diseases have killed tens of millions of people over the centuries, and to date there are still some difficulties in controlling infectious diseases. Among the researches for the inhibition of infectious diseases, the discovery of penicillin has become a turning point in inhibiting or treating pathogenic microorganisms caused by antibiotics, inhibiting disease and improving health health and prolonging life span. However, direct or indirect damages caused by pathogenic microorganisms still cause various problems and diseases in economy, environment, medicine and health.
특히, 결핵은 기원전 7천년 경 석기 시대의 화석에서 그 흔적이 발견된 이래 인류의 생명을 가장 많이 앗아간 전염성 질병이다. 결핵은 1882년 독일의 의사이자 세균학자 하인리히 헤르만 로베르트 코흐(Heinrich Hermann Robert Koch, 1843~1910)가 결핵(Tuberculosis)의 병원체인 결핵균(mycobacterium tuberculosis; MTB)을 발견함으로써 세상에 알려지게 되었으며 현재까지도 아시아, 아프리카, 남미의 저개발 국가를 비롯한 전 세계에 걸쳐 분포하고 있으며 감염의 위험성을 유발하고 있다. 결핵은 결핵균의 감염에 의해 유발되는 전염성 질환으로 사람과 사람간의 접촉을 통해서 전파되며 결핵환자의 기침과 비말에 의해 감염된다. 세계보건기구(WHO)의 보고에 따르면 현재 전 세계인구의 1/3 정도가 결핵균에 감염되어 있으며 매년 150만 명의 환자가 결핵으로 사망하고 있다고 보고하였다. 또한 2013년 네이처 저널에 따르면 1990년을 기준으로 AIDS 환자의 결핵 감염률(Co-infection)이 점차적으로 증가하고 있다고 보고되었다. In particular, tuberculosis is an infectious disease that has taken the greatest number of lives since the discovery of traces in the fossil of the Stone Age around 7000 BCE. Tuberculosis became known to the world in 1882 when the German physician and bacteriologist Heinrich Hermann Robert Koch (1843-1910) discovered mycobacterium tuberculosis (MTB), a pathogen of tuberculosis, It is distributed throughout the world, including underdeveloped countries in Africa and South America, and is causing the risk of infection. Tuberculosis is a communicable disease caused by infection of Mycobacterium tuberculosis. It spreads through contact between humans and humans and is infected by cough and droplets of tuberculosis patients. According to a report by the World Health Organization (WHO), about one-third of the world's population is now infected with tuberculosis, and 1.5 million people die annually from tuberculosis. According to the Nature Journal in 2013, the rate of tuberculosis infection (Co-infection) in AIDS patients is gradually increasing in 1990.
현재 임상에서 결핵 치료제로 사용되고 있는 주요 치료약물로 Rifampicin, Isoniazid, Ethambutol, Pyrazinamide와 같은 항결핵 약물이 사용되고 있으나 대부분 1970년 이전에 개발된 것이며 장기간 복용할 경우 심각한 약물부작용(Rifampicin; 정신착란, 경련, 위장장애, 복통, 백혈구감소, 고혈당, 간장애, 신부전, 혈뇨, 단백뇨, 호흡곤란, 천식성 발작, Isoniazid; 간염, 혈관염, 경련, 말초신경병증, 지각이상, 고혈당증, 신부전, 배뇨곤란, Ethambutol; 시신경염, 시력장애, 환각, 간장애, 소화기장애, Pyrazinamide; 간염, 황달, 간장애, 고요산혈증, 발열)이 유발되고 있으며 이와 동시에 돌연변이를 통한 약제내성이 야기되고 있다. 특히, Rifampicin과 Isoniazid에 내성을 나타내는 다제내성 결핵균(multidrug-resistant tuberculosis, MDR-TB)의 경우 기존 1차 항결핵제 투여로는 치료가 될 수 없으며, MDR-TB의 치료를 위한 2차 항결핵제 및 복합 병용투여는 심각한 부작용을 동반하여 환자의 건강회복에 다양한 문제를 일으키고 있으며 이러한 다제내성 결핵균은 국가적으로 보건향상과 국민건강에 많은 문제를 초래하고 있다. 최근 의료 진단기술과 치료기술의 발전에도 불구하고 다이어트로 면역이 저하된 20대~30대의 젊은 세대와 60대 이상의 노년층에서 잠복감염 및 만성감염을 통해 새로운 결핵 환자가 증가되고 있는 실정이다. Currently, anti-tuberculosis drugs such as Rifampicin, Isoniazid, Ethambutol, and Pyrazinamide have been used in clinical trials, but most of them have been developed before 1970. Rifampicin (long-term use) Nephropathy, nephropathy, peripheral neuropathy, nephropathy, hyperglycemia, renal failure, dysuria, Ethambutol, optic neuritis, neurodegenerative diseases, neurodegenerative disorders, , Vision impairment, hallucinations, liver disorders, gastrointestinal disorders, pyrazinamide, hepatitis, jaundice, liver disorder, hyperuricemia, fever) and at the same time mutagenic drug resistance. In particular, multidrug-resistant tuberculosis (MDR-TB), which is resistant to rifampicin and isoniazid, can not be treated with the first-line antituberculous drug. Second-line anti-tuberculosis drugs Administration has been associated with serious side effects and has caused various problems in the recovery of the patients' health. These multidrug-resistant Mycobacterium tuberculosis are causing national health improvement and public health problems. Despite the recent advances in medical diagnostic technology and therapeutic technology, new tuberculosis patients are increasing in the 20s and 30s, who have been immunized by diet, through latent infections and chronic infections.
이러한 이유로, 보다 효과적이고 부작용이 적은 새로운 항결핵제의 개발과 보급이 절실히 요구되고 있다. 현재 전 세계적으로 다국적 제약기업을 중심으로 다양한 결핵 치료제 개발이 활발히 진행되고 있으나 아직 기존 치료약물을 대체할 수 있는 효과적인 항결핵제가 개발되지 않고 있다. For this reason, the development and dissemination of new anti-tuberculosis drugs which are more effective and have less side effects are urgently required. Currently, various TB drugs are actively developed around multinational pharmaceutical companies around the world, but effective anti-tuberculosis drugs that can replace existing therapeutic drugs have not been developed yet.
또한, 결핵은 사람뿐만 아니라 소, 염소, 돼지, 조류를 포함하는 다양한 동물에도 심각한 전염성 질병으로 축산업에 있어서도 매우 심각한 가축전염병성 질병이다. 특히, 소결핵은 결핵균이 소의 호흡기나 소화기로 들어와 발병하며 결핵에 감염된 소의 침이나 배설물을 통해 사람에게 감염될 수 있다. 현재 소결핵에 감염된 가축의 치료제는 사람에 사용하는 Rifampicin, Isoniazid 등의 항결핵 약물이 사용되고 있지만 감염된 가축은 사람으로의 전염을 차단하기 위해 전부 도살처리 되고 있는 실정이다. 이러한 이유로 항결핵제는 인간뿐만 아니라 축산업에 있어서도 매우 중요한 역할을 하고 있는 약물이다. Tuberculosis is also a serious livestock epidemic disease in the livestock industry due to serious infectious diseases not only in humans but also in various animals including cattle, goats, pigs and birds. In particular, bovine tuberculosis can develop into a respiratory or digestive tract of bovine tuberculosis, and infect people through tuberculosis infected cattle saliva or excreta. Currently, anti-tuberculosis medicines such as Rifampicin and Isoniazid, which are used in humans, are used for treatment of livestock infected with bovine tuberculosis. However, infected livestock is totally slaughtered to prevent transmission to human. For this reason, anti-tuberculosis drug plays a very important role not only in human beings but also in animal husbandry.
현재 사용되고 있는 대부분의 항결핵제는 화학적인 합성을 통해 제조된 것으로서 상기와 같은 내성 및 부작용을 유발하는 등의 많은 한계를 나타내고 있다. 따라서 최근에는 동물, 식물, 미생물 및 다양한 한약재와 약용식물을 포함하는 천연물로부터 새로운 항결핵 활성물질을 분리, 개발하고자 하는 연구가 활발하게 진행되고 있으며 자연친화적 물질의 개발이 중요한 과제로 부각되고 있다. 하지만, 이러한 새로운 항균물질의 연구와 개발이 쉽지만은 않은 실정이다. 새로운 항균물질을 개발하기 위해서는 항균효과가 광범위하고 기존 내성 균주에도 효과를 나타내며 잠복감염 및 만성감염에도 효과를 나타내고 장기간 투여하여도 부작용 없이 안전하여야 한다는 점 등이 고려되어야 한다. Most of the currently used anti-tuberculosis agents are produced through chemical synthesis, and thus exhibit many limitations such as causing the above-mentioned resistance and side effects. Therefore, in recent years, researches for separating and developing new anti-tuberculosis active materials from natural products including animals, plants, microorganisms and various herbal medicines and medicinal plants have been actively carried out, and the development of environmentally friendly substances has become an important issue. However, research and development of these new antimicrobial substances is not easy. In order to develop a new antimicrobial substance, it should be considered that the antimicrobial effect is wide, it has an effect on the existing resistant strains, has an effect on latent infection and chronic infection, and should be safe even without long term administration.
이러한 관점에서, 10E,12Z 공액리놀레산은 자연에 존재하는 다양한 종류의 동식물에 함유되어 있는 천연물-유래 활성 지방산 또는 생리활성물질로서 항산화, 항알러지, 항당뇨 효과, 콜레스테롤 감소, 항염증, 항암 효과 및 심근경색, 동맥경화 억제와 같은 다양한 약리작용과 효과가 보고되었다. 또한 체지방 감소, 성장촉진 등의 활성을 갖고 있는 것으로 보고되고 있으며, 식물과 해양생물에 다량 함유되어 있으며 10E,12Z 공액리놀레산은 급성독성과 만성독성이 없는 안전한 화합물이다. In view of this, 10E, 12Z conjugated linoleic acid is a natural product-derived active fatty acid or a physiologically active substance contained in various kinds of animals and plants existing in nature and has antioxidant, antiallergic, antidiabetic effect, cholesterol reduction, anti- Myocardial infarction, and inhibition of arteriosclerosis have been reported. It is reported to have activity such as decrease of body fat and promotion, and it is contained in large amount in plants and marine life. 10E, 12Z conjugated linoleic acid is a safe compound without acute toxicity and chronic toxicity.
세계보건기구(WHO)가 지정한 세계 감염성 질병중의 하나인 말라리아의 치료 약물인 Artemisinin과 Artesunate는 생약인 개똥쑥에서 추출된 생리활성물질이다. 또한, Artemisinin-내성 말라리아를 치료하기 위한 Artemisinin based Combination Therapy(ACT)에도 Artemisinin과 Artesunate가 기본약물로 사용되고 있다. 최근, 국내 제약사에서 말라리아 신약을 연구, 개발하여 WHO에 공급하고 있으며 아프리카와 국내에도 판매하고 있다. Artemisinin and Artesunate, the therapeutic drugs of malaria, one of the World Infectious Diseases designated by the World Health Organization (WHO), are physiologically active substances extracted from the herbal medicine dog mugwort. Artemisinin-based Combination Therapy (ACT) for the treatment of Artemisinin-resistant malaria also uses Artemisinin and Artesunate as basic drugs. Recently, Korean pharmaceutical companies research and develop new malaria drugs, supply them to WHO, and sell them to Africa and Korea.
이러한 배경 하에서, 본 발명자는 천연물-유래 항균활성물질을 연구하여 개발하기위해 노력하던 중 지방산으로 알려진 Conjugated(10E,12Z)-Linoleic acid가 결핵균의 분열증식과 생존을 억제하고 결핵균의 성장을 저해하는 높은 항결핵균 활성과 결핵균에 대한 항균효과를 나타낸다는 것을 확인함으로써 본 발명을 완성하게 되었다.
Under these circumstances, the present inventors have made efforts to study and develop natural products-derived antimicrobial active substances, and conjugated (10E, 12Z) -Linoleic acid, known as fatty acid, has been used to inhibit the growth and proliferation of Mycobacterium tuberculosis, The present invention has been accomplished by confirming that it exhibits anti-Mycobacterium tuberculosis activity and antibacterial effect against Mycobacterium tuberculosis.
본 발명의 목적은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 치료, 개선 또는 예방용 약학 조성물을 제공하는 것이다. It is an object of the present invention to provide a pharmaceutical composition for the treatment, improvement or prevention of tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient.
본 발명의 다른 하나의 목적은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 예방 또는 개선용 건강식품 조성물을 제공하는 것이다. Another object of the present invention is to provide a health food composition for preventing or ameliorating tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient.
본 발명의 또 다른 하나의 목적은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 예방 또는 개선용 사료 조성물을 제공하는 것이다. It is still another object of the present invention to provide a feed composition for preventing or improving tuberculosis comprising 10E, 12Z conjugated linoleic acid as an active ingredient.
또한, 본 발명의 다른 목적은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 치료, 개선 또는 예방용 동물의약품 조성물을 제공하는 것이다. Another object of the present invention is to provide an animal pharmaceutical composition for the treatment, improvement or prevention of tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient.
또한, 본 발명의 또 다른 목적은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 예방 또는 개선용 의약외품 조성물을 제공하는 것이다.
Still another object of the present invention is to provide a quasi-drug composition for preventing or ameliorating tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient.
상기 목적을 달성하기 위한 하나의 양태로서, 본 발명은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 치료, 개선 또는 예방용 약학적 조성물을 제공한다. In one aspect of the present invention, the present invention provides a pharmaceutical composition for treating, improving or preventing tuberculosis comprising 10E, 12Z conjugated linoleic acid as an active ingredient.
본 발명에서 사용되는 용어, "Conjugated(10E,12Z)-Linoleic acid"는 "(10E,12Z)-10,12-Octadecadienoic acid" 또는 "Linoleic acid(10-trans,12-cis)" 또는 "10E,Z12-CLA" 라고도 불리며 실온에서 무색의 용액으로 화학구조식은 C18H32O2 이고 몰 질량(Molar mass)은 280.45 g/mole 이다. The term " Conjugated (10E, 12Z) -Linoleic acid "as used in the present invention refers to" 10E, 12Z) -10,12-Octadecadienoic acid "or" Linoleic acid (10- , Z12-CLA ". It is a colorless solution at room temperature. It has a chemical formula of C 18 H 32 O 2 and a molar mass of 280.45 g / mole.
본 발명에서는 10E,12Z 공액리놀레산을 에탄올로 녹인 후 배지를 사용하여 농도별로 용액을 제조하고 액체배지 발색법(Resazurin microtiter assay)을 사용하여 결핵균에 대한 시험물질의 항균활성을 측정하였다. 본 발명의 일실시 예에서, 10E,12Z 공액리놀레산은 결핵균에 대한 항균활성을 나타내고 있음이 확인되었다 (도 1). In the present invention, 10E, 12Z conjugated linoleic acid was dissolved in ethanol, and a solution was prepared for each concentration by using a medium. The antimicrobial activity of a test substance against Mycobacterium tuberculosis was measured using a Resazurin microtiter assay. In one embodiment of the present invention, 10E, 12Z conjugated linoleic acid showed antibacterial activity against Mycobacterium tuberculosis (Fig. 1).
본 발명에서는 10E,12Z 공액리놀레산을 에탄올로 녹인 후 배지를 사용하여 농도별로 용액을 제조하고 MGIT SYSTEM 측정법을 사용하여 결핵균에 대한 시험물질의 항균활성 또는 항균효과를 측정하였다. 본 발명의 일실시 예에서, 10E,12Z 공액리놀레산은 결핵균에 대한 항균활성 또는 항균효과를 나타내고 있음이 확인되었다 (도 2). In the present invention, 10E, 12Z conjugated linoleic acid was dissolved in ethanol, the solution was prepared by using the culture medium, and the antimicrobial activity or the antibacterial effect of the test substance against Mycobacterium tuberculosis was measured using the MGIT SYSTEM measurement method. In one embodiment of the present invention, it was confirmed that 10E, 12Z conjugated linoleic acid exhibited antibacterial activity or antibacterial effect against Mycobacterium tuberculosis (Fig. 2).
본 발명에서 사용되는 용어 "결핵균(mycobacterium tuberculosis)"는 인형(人型)결핵균, 우형(牛型)결핵균, 서형(鼠型)결핵균, 조형(鳥型菌)결핵균으로 분류되며 보통, 길이 1.2~4.0m, 폭 0.3~0.5m 정도의 아주 미세한 막대 모양으로 된 간균 형태의 호기성 그램 양성균 이다. 또한, 인형결핵균은 A형, I형, B형, C형의 4가지로 분류되며 pH 6.8~7.0, 온도는 37가 최적의 생장조건이다. The term " mycobacterium tuberculosis "as used in the present invention is classified as a human tubercle bacillus, a bovine type tubercle bacillus, a western type tubercle bacillus, and a bird type bacillus. It is an aerobic gram-positive bacterium in the form of a very fine rod-shaped bacterium with a width of 4.0 m and a width of 0.3 to 0.5 m. Mycobacterium tuberculosis is classified into four types A, I, B and C, pH 6.8 ~ 7.0 and temperature 37 is the optimal growth condition.
본 발명에서 사용되는 용어 "결핵(tuberculosis)"은 결핵균 감염 환자로부터 호흡기를 통해 나온 미세한 침방울 혹은 비말에 의해 직접 감염되며 그 정의상 결핵균에 의한 감염 때문에 발생하는 질환이다. 현재, 결핵균은 공기를 통해서 감염되기 때문에 폐 조직에서 결핵이 잘 생긴다. 따라서 보통 결핵이라고 하면 폐결핵을 의미하지만 결핵균은 폐 이외에도 흉막, 림프선, 척추, 뇌, 신경, 신장, 위장관, 생식기, 뼈, 근육 등 대부분의 조직이나 장기에도 침입해 증상을 일으킬 수 있다. As used herein, the term "tuberculosis" is a disease caused directly by a fine needle or droplet through a respiratory tract from a patient with M. tuberculosis infection, and is by definition an infection caused by M. tuberculosis. Currently, tubercle bacillus is well-developed in lung tissue because it is transmitted through air. Thus, tuberculosis usually refers to pulmonary tuberculosis. However, tuberculosis can invade most tissues and organs such as pleura, lymph node, spine, brain, nerve, kidney, gastrointestinal tract, genitalia, bone and muscle as well as lung.
결핵은 결핵균에 감염된 장기에 따라서 폐결핵, 고관절결핵, 골결핵, 골반결핵, 고환결핵, 방광결핵, 신장결핵, 장결핵, 피부결핵, 안결핵, 후두결핵, 림프절결핵, 척추결핵, 결핵성 뇌막염 같은 중추신경계 결핵, 결핵성 복막염 또는 결핵성 난관염 등으로 다양하게 분류되며 만성화에 따라 만성결핵 또는 잠복결핵으로 나뉜다. 결핵균은 숙주의 몸속 영양분을 이용해 천천히 증식하며 결핵에 걸리면 기운이 없고 쉽게 피로를 느끼며 체중이 감소하는 등의 증상이 주로 나타난다. 또한 결핵균에 감염된 장기에 따라서 증상이 다르게 나타나는데 폐결핵의 경우에는 기침과 가래, 흉통 등의 증상이 생기며 비뇨 생식계 결핵에서는 빈뇨, 배뇨곤란, 혈뇨 등이 나타나고 골관절 결핵에서는 해당 부위 뼈에 통증이 유발되며 림프선 결핵이면 전신, 특히 목 위나 겨드랑이의 림프절이 커지면서 통증과 압박감이 유발된다. 또한 척추 결핵의 경우 허리통증을, 중추신경계 결핵에서는 두통과 구토 등의 증상이 나타난다. 사람의 몸이 결핵균에 감염되면 면역세포와의 염증반응에 의해 아주 느린 속도로 몸의 정상조직을 파괴시키며 치즈 같은 형태의 고름이 형성되고 그 주위에 육아종(Granuloma, 덩어리로 된 혹)이 발생하게 된다. Tuberculosis may be caused by tuberculosis or tuberculosis according to organs such as tuberculosis, hip tuberculosis, bone tuberculosis, pelvic tuberculosis, testicular tuberculosis, bladder tuberculosis, renal tuberculosis, tuberculosis, skin tuberculosis, tuberculosis, laryngeal tuberculosis, tuberculosis, Tuberculosis, tuberculous peritonitis, tuberculous ovarian tuberculosis, and chronic tuberculosis. Mycobacterium tuberculosis grows slowly using the nutrients in the body of the host, and when it is caught in tuberculosis, symptoms such as lack of energy, easy fatigue, and decrease in body weight appear. In addition, symptoms vary according to the organ that is infected with Mycobacterium tuberculosis. In pulmonary tuberculosis, symptoms such as cough, sputum and chest pain occur, and urinary frequency, urination difficulty and hematuria occur in tuberculosis of urography. In osteoarthritic tuberculosis, If tuberculosis, the lymph nodes of the whole body, especially the neck or armpit, grow, causing pain and pressure. In addition, back pain in patients with spinal tuberculosis, central nervous system tuberculosis symptoms such as headache and vomiting appear. When a person's body is infected with Mycobacterium tuberculosis, the normal tissue of the body is destroyed at a very slow rate by the inflammatory reaction with immune cells, and a cheese-like pus is formed and granuloma (lumps) do.
본 발명에서 사용되는 "항균"이란 세균에 저항하는 효능, 보다 상세하게는 약제나 생리활성물질 또는 알려진 화학물질에 의한 세균 및 진균의 생육저해작용을 의미하는 것으로서 본 발명의 목적상 상기 결핵균의 분열증식 억제 및 성장저해작용을 의미한다. The term " antibacterial "as used in the present invention means an effect against bacteria, more specifically, an effect of inhibiting the growth of bacteria and fungi by a drug, a physiologically active substance or a known chemical substance. For the purpose of the present invention, Inhibiting growth and inhibiting growth.
본 발명의 약학적 조성물로 치료, 개선 또는 예방이 가능한 결핵은 이에 제한되지는 않으나, 바람직하게는 폐결핵일 수 있으며, 보다 바람직하게는 결핵균(Mycobacterium tuberculosis) 감염에 의하여 발생하는 감염성 결핵일 수 있다. The tuberculosis that can be treated, improved or prevented by the pharmaceutical composition of the present invention is not limited thereto, but it may be preferably pulmonary tuberculosis, more preferably infectious tuberculosis caused by infection with Mycobacterium tuberculosis.
본 발명에서 상기 약학적 조성물의 결핵의 치료, 개선 또는 예방 효과는 결핵균의 분열증식 및 성장을 저해하는 항균활성 또는 항균효과를 가짐으로써 달성되는 것일 수 있다. 본 발명의 일실시 예에서는 액체배지 발색법(Resazurin microtiter assay)과 MGIT SYSTEM 측정법을 통하여 10E,12Z 공액리놀레산이 결핵균의 성장 및 분열증식을 저해하는 항균활성 또는 항균효과를 가지고 있음을 확인함으로써 결핵의 치료, 개선 또는 예방효과를 달성할 수 있음을 확인하였다(도 1 내지 도 2). In the present invention, the therapeutic, ameliorative or preventive effect of the pharmaceutical composition on tuberculosis may be achieved by having antimicrobial activity or antibacterial effect which inhibits proliferation and growth of Mycobacterium tuberculosis. In one embodiment of the present invention, 10E, 12Z conjugated linoleic acid was found to have antimicrobial activity or antimicrobial effect inhibiting the growth and proliferation of Mycobacterium tuberculosis through the Resazurin microtiter assay and the MGIT SYSTEM assay, Treatment, improvement or preventive effect can be achieved (Figs. 1 to 2).
본 발명의 10E,12Z 공액리놀레산은 이에 제한되지는 않으나, 바람직하게는 화학적 합성에 의해 얻어지는 합성물질 또는 동물, 식물 또는 자연물에서 유래되는 물질일 수 있다. 10E,12Z 공액리놀레산은 업계에 공지된 일반적인 합성방법, 동물, 식물 또는 자연물에서 유래되는 추출방법이나 분리방법 또는 정제방법을 이용하여 얻어질 수 있다. 또한, 상기 추출방법은 이에 제한되지는 않으나, 바람직하게는 유기용매 추출, 열탕 추출, 열수 추출, 냉침 추출, 증기 환류 추출, 환류 냉각 추출, 초음파 추출, 진공농축 또는 동결건조 등의 방법을 사용할 수 있다.The 10E, 12Z conjugated linoleic acid of the present invention is not limited thereto, but may preferably be a synthetic material obtained by chemical synthesis or a material derived from an animal, a plant or a natural product. 10E, 12Z conjugated linoleic acid can be obtained by an ordinary extraction method, a separation method, or a purification method derived from synthetic methods, animals, plants or natural products known in the art. The extraction method may be, but not limited to, organic solvent extraction, hot water extraction, hot water extraction, cold extraction, steam reflux extraction, reflux cooling extraction, ultrasonic extraction, vacuum concentration or freeze- have.
본 발명에서 사용되는 용어, "예방"은 상기 조성물의 섭취 또는 투여로 상기 질환의 발병을 억제 또는 지연시키는 모든 행위를 의미하며 "치료"는 상기 조성물의 섭취 또는 투여로 상기 질환의 증세가 호전되거나 이롭게 되는 모든 행위를 의미한다. As used herein, the term "prevention" means any action that inhibits or delays the onset of the disease by ingestion or administration of the composition, and "treatment" means that the ingestion or administration of the composition It means all acts that are benefited.
본 발명에서 사용되는 용어, "개선"이란 10E,12Z 공액리놀레산을 유효성분으로 포함하는 조성물을 섭취 또는 투여함으로서 상기 질환의 발병 또는 증상을 완화시키거나 결핵균의 성장 및 분열증식을 억제 또는 감소시킴으로써 결핵의 증상을 완화시키는 효과를 의미한다. As used herein, the term "improvement" refers to the administration of a composition comprising 10E, 12Z conjugated linoleic acid as an active ingredient to alleviate the onset or symptom of the disease or to inhibit or reduce the growth and proliferation of Mycobacterium tuberculosis, Of the symptoms.
본 발명의 약학적 조성물은 약제학적으로 허용 가능한 담체를 포함할 수 있으며 이에 제한되지는 않으나, 바람직하게는 다음과 같다. The pharmaceutical composition of the present invention may include, but is not limited to, a pharmaceutically acceptable carrier.
본 발명의 약학 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 덱스트로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트 및 광물유를 들 수 있다. 약학적으로 허용 가능한 담체를 포함하는 조성물은 경구 또는 비경구의 여러 가지 제형일 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제될 수 있다. 경구투여를 위한 고형제제에는 정제환제, 산제, 과립제, 캡슐제 등이 포함될 수 있으며 이러한 고형제제는 하나 이상의 화합물에 적어도 하나 이상의 부형제, 예를 들면, 전분, 탄산칼슘, 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제될 수 있다. 또한, 단순한 부형제 이외에 스테아린산 마그네슘, 탈크 등과 같은 윤활제들도 사용될 수 있다. 경구투여를 위한 액상제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 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, 아미노산제, 유제, 동결건조제제, 좌제가 포함될 수 있다. 비수성용제, 현탁용제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테로 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로젤라틴 등이 사용될 수 있다. Examples of carriers, excipients and diluents that can be included in the pharmaceutical composition of the present invention include dextrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, calcium silicate, cellulose, methylcellulose, microcrystalline cellulose, poly Vinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, and mineral oil. Compositions comprising a pharmaceutically acceptable carrier can be of various oral or parenteral formulations. In the case of formulation, it can be prepared using diluents or excipients such as fillers, extenders, binders, humectants, disintegrants, surfactants and the like which are usually used. Solid formulations for oral administration may include tablet pills, powders, granules, capsules, and the like, which may contain one or more excipients, such as starch, calcium carbonate, sucrose, or lactose lactose, gelatin, and the like. In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions, syrups and the like. Various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included in addition to water and liquid paraffin, which are simple diluents commonly used. have. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, amino acids, emulsions, lyophilized formulations, and suppositories. Examples of the non-aqueous solvent and the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, and injectable ester such as ethyl oleate. Examples of the suppository base include witepsol, macrogol, tween 61, cacao paper, laurin, glycerogelatin and the like.
또한, 본 발명의 약학적 조성물은 정제, 환제, 산제, 과립제, 캡슐제, 서방형제제, 설하정, 현탁제, 내용액제, 유제, 주사제, 주정제, 경피흡수제, 점막흡수제, 구강붕해제, 패치제, 필름제, 도포제, 첩부제, 흡입제, 시럽제, 에멀젼, 겔제, 연고제, 점안제, 멸균된 수용액, 비수성용제, 동결건조제제, 좌제 또는 에어로졸로 이루어진 군으로부터 선택되는 어느 하나의 제형을 가질 수 있으며, 상기 예에 의해 제조 가능한 제형의 형태가 제한되는 것은 아니다. In addition, the pharmaceutical composition of the present invention may be in the form of tablets, pills, powders, granules, capsules, sustained release formulations, suspensions, suspensions, solutions, emulsions, injections, It may have any one form selected from the group consisting of a patch, a film, a coating agent, a patch, an inhalant, a syrup, an emulsion, a gel, an ointment, eye drops, a sterilized aqueous solution, a nonaqueous agent, a lyophilized agent, a suppository or an aerosol And the form of the formulations which can be produced by the above examples is not limited.
본 발명의 약학적 조성물은 그 투여용량에 특별한 제약은 없고, 체내 흡수도, 체중, 환자의 연령, 성별, 건강상태, 식이, 투여시간, 투여방법, 배설율 및 질환의 중증도 등에 따라 변화될 수 있다. 본 발명의 약학적 조성물은 유효량 범위를 고려하여 제조하도록 하며, 이렇게 제형화된 단위 투여형 제제는 필요에 따라 전문가의 판단과 개인의 요구에 따라 전문화된 투약법을 사용하거나 일정 시간의 간격으로 수회 투여할 수 있다. The dosage form of the pharmaceutical composition of the present invention is not particularly limited and may be varied depending on the degree of absorption, body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, have. The pharmaceutical composition of the present invention is prepared in consideration of an effective dose range, and the formulated unit dosage form can be prepared by using a specialized dosage regimen according to the judgment of the expert and the needs of the individual, Lt; / RTI >
다른 하나의 양태로서, 본 발명은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 예방 또는 개선용 건강식품 조성물을 제공한다. 상기 건강식품 조성물은 바람직하게는 결핵균에 대한 항균활성 또는 항균효과를 가짐으로써 달성되는 것일 수 있으며, 이에 대해서는 상기에서 설명한 바와 같다. In another aspect, the present invention provides a health food composition for preventing or ameliorating tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient. The health food composition may preferably be obtained by having an antibacterial activity or antibacterial effect against Mycobacterium tuberculosis, as described above.
본 발명의 조성물을 식품의 첨가물로 사용할 경우, 상기 조성물을 그대로 첨가하거나 다른 건강식품 또는 그 성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용할 수 있다. 유효성분의 혼합량은 사용목적에 따라 적합하게 결정될 수 있으며, 본 발명의 식품 조성물은 혼합량에 큰 제한은 없다. When the composition of the present invention is used as an additive for food, the composition may be added as it is or may be used together with other health food or ingredients thereof, and may be suitably used according to a conventional method. The mixing amount of the active ingredient can be suitably determined according to the purpose of use, and the amount of the food composition of the present invention is not particularly limited.
본 발명의 식품의 종류에는 특별한 제한은 없으며 상기 조성물을 첨가할 수 있는 건강식품의 예로는 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 쥬스, 드링크제, 알코올성 음료 및 비타민 복합제 등이 있고 통상적인 의미에서의 건강식품을 모두 포함할 수 있으며, 동물을 위한 사료로 이용되는 식품을 포함할 수 있다. The food of the present invention is not particularly limited and examples of the health food to which the composition can be added include meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gum, Dairy products, various soups, beverages, tea, juice, drinks, alcoholic beverages and vitamin complexes, and may include foods used as food for animals, which may include all health foods in a conventional sense.
또한, 본 발명의 건강식품 조성물이 음료의 형태로 사용될 경우에는 통상의 음료와 같이 여러 가지 감미제, 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상기 외에 본 발명의 건강식품 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 함유할 수 있다. 그밖에 천연 과일음료, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 본 발명의 조성물을 차, 쥬스 및 드링크의 형태로 제조하여 음용하도록 하거나 과립화, 정제화, 캡슐화 및 분말화하여 섭취할 수 있으나, 상기 예에 의해 제조 가능한 형태가 제한되는 것은 아니다. In addition, when the health food composition of the present invention is used in the form of a drink, it may contain various sweeteners, flavors, or natural carbohydrates as an additional ingredient such as ordinary beverages. In addition to the above, the health 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 thickening agents, pH adjusting agents, stabilizers, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, it may contain flesh for the production of natural fruit drinks, fruit juice drinks and vegetable drinks. The composition of the present invention can be prepared in the form of tea, juice, and drink, and then consumed for drinking, granulated, tableted, encapsulated, and powdered, but the form that can be manufactured by the above example is not limited.
또 다른 하나의 양태로서, 본 발명은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 예방 또는 개선용 사료 조성물을 제공한다. 상기 사료 조성물은 바람직하게는 결핵균에 대한 항균활성 또는 항균효과를 가짐으로써 달성되는 것일 수 있으며, 이에 대해서는 상기에서 설명한 바와 같다. In another aspect, the present invention provides a feed composition for preventing or improving tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient. The feed composition may preferably be obtained by having an antibacterial activity or an antibacterial effect against Mycobacterium tuberculosis, as described above.
본 발명의 조성물을 사료 첨가물로 사용할 경우, 상기 조성물을 그대로 첨가하거나 다른 사료 성분과 함께 혼합하여 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용할 수 있다. 유효성분의 혼합량은 사용목적에 따라 적합하게 결정될 수 있으며 본 발명의 사료 조성물은 혼합량에 큰 제한은 없다. When the composition of the present invention is used as a feed additive, the composition may be added as it is or may be mixed with other feed ingredients and used appropriately according to a conventional method. The amount of the active ingredient to be mixed may be appropriately determined according to the purpose of use, and the amount of the feed composition of the present invention is not particularly limited.
본 발명의 사료의 종류에는 특별한 제한은 없으며 상기 조성물은 보조 성분으로 아미노산, 무기염류, 비타민, 항산화, 미생물 제제 등과 같은 각종 보조제 및 분쇄 또는 파쇄된 밀, 보리, 옥수수 등의 식물성 단백질사료, 혈분, 육분, 생선분 등의 동물성 단백질 사료, 동물성 지방 및 식물성 지방과 같은 주성분 이외에도 영양 보충제, 성장 촉진제, 소화 흡수 촉진제, 질병 예방제와 함께 사용될 수 있다. 상기 사료의 투여 형태, 방법 및 제형은 다양하게 할 수 있으며, 상기 예에 의해 제조 가능한 형태가 제한되는 것은 아니다. There is no particular limitation on the kind of the feed of the present invention, and the composition may be supplemented with various auxiliaries such as amino acids, inorganic salts, vitamins, antioxidants, microbial preparations and the like and pulverized or crushed wheat, vegetable protein feeds such as barley, corn, Can be used in combination with nutritional supplements, growth promoters, digestion-absorption promoters, and disease prevention agents in addition to the main components such as animal protein feeds, animal fat and vegetable fat. The dosage form, method and formulation of the feed can be varied, and the form that can be produced by the example is not limited.
본 발명의 사료 조성물은 포유류, 가금류, 어류 및 갑각류를 포함하는 다수의 동물식이 즉, 사료에 적용할 수 있다. 상업용 가축으로 중요한 소, 돼지, 말, 사슴, 염소 등의 가축을 포함하는 포유류, 코끼리, 낙타, 기린, 원숭이 등의 동물원 동물, 개, 고양이 등의 애완동물에게 사용할 수 있으며, 닭, 오리, 거위, 비둘기 등의 가금류 및 붕어, 잉어, 송어, 숭어, 새우와 같은 상업적으로 사육되는 어류 및 갑각류에 사용할 수 있으나, 이에 제한되지는 않는다. The feed composition of the present invention can be applied to a large number of animal diets, i.e. feeds, including mammals, poultry, fish and crustaceans. Commercial livestock can be used for zoo animals such as mammals, elephants, camels, giraffes, monkeys, pets such as cows, pigs, horses, deer and goats; pets such as dogs and cats; , Pigeons, and commercially available fish and crustaceans such as carp, carp, trout, mullet, and shrimp.
또한, 다른 하나의 양태로서, 본 발명은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 치료, 개선 또는 예방용 동물의약품 조성물에 관한 것이다. 즉, 본 발명의 조성물은 결핵의 치료, 개선 또는 예방을 목적으로 동물의약품 조성물로 사용될 수 있다. In another aspect, the present invention relates to an animal pharmaceutical composition for the treatment, improvement or prevention of tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient. That is, the composition of the present invention can be used as an animal pharmaceutical composition for the purpose of treatment, improvement or prevention of tuberculosis.
상기 조성물은 바람직하게는 결핵균에 대한 항균활성 또는 항균효과를 가짐으로써 달성되는 것일 수 있으며, 이에 대해서는 상기에서 설명한 바와 같다. The composition may preferably be achieved by having an antibacterial activity or an antibacterial effect against Mycobacterium tuberculosis, as described above.
본 발명의 조성물을 동물의약품 조성물로 사용할 경우, 상기 조성물을 그대로 사용하거나 다른 의약품 또는 의약외품 성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있으며 이에 제한되는 것은 아니다. 유효성분의 혼합양은 사용 목적(예방, 건강, 개선 또는 치료적 처치)에 따라 적합하게 결정될 수 있다. When the composition of the present invention is used as an animal pharmaceutical composition, the composition may be used as it is or may be used together with other pharmaceutical or quasi-drug components, and may be suitably used according to a conventional method, but is not limited thereto. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health, improvement or therapeutic treatment).
또한, 다른 하나의 양태로서, 본 발명은 10E,12Z 공액리놀레산을 유효성분으로 포함하는 결핵의 예방 또는 개선용 의약외품 조성물에 관한 것이다. 즉, 본 발명의 조성물은 결핵의 예방 또는 개선을 위한 의약외품 조성물에 첨가될 수 있다. In another aspect, the present invention relates to a quasi-drug composition for preventing or improving tuberculosis containing 10E, 12Z conjugated linoleic acid as an active ingredient. That is, the composition of the present invention can be added to a quasi-drug composition for preventing or improving tuberculosis.
상기 조성물은 바람직하게는 결핵균에 대한 항균활성 또는 항균효과를 가짐으로써 달성되는 것일 수 있으며, 이에 대해서는 상기에서 설명한 바와 같다. The composition may preferably be achieved by having an antibacterial activity or an antibacterial effect against Mycobacterium tuberculosis, as described above.
본 발명의 조성물을 의약외품 첨가물로 사용할 경우, 상기 조성물을 그대로 첨가하거나 다른 의약외품 또는 의약외품 성분과 함께 사용할 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있으며 이에 제한되는 것은 아니다. 유효성분의 혼합양은 사용 목적(예방, 건강, 또는 개선)에 따라 적합하게 결정될 수 있다. 본 발명의 의약외품 조성물의 종류에는 제한은 없으나 바람직하게는 소독 청결제, 샤워폼, 가그린, 물티슈, 세제비누, 스프레이, 핸드워시, 연고제, 도포제, 크림제, 카타플라스마제, 가습기 충진제, 화장품, 샴푸, 코팅제, 세정제, 여성 청결제, 위생패드, 생리대, 콘돔, 밴드, 필터 충진제, 위생장갑, 위생용기, 위생봉투 또는 마스크의 제조에 사용될 수 있으며, 상기 예에 의해 제조 가능한 형태가 제한되는 것은 아니다.
When the composition of the present invention is used as a quasi-drug additive, the composition may be added as it is or may be used together with other quasi-drugs or quasi-drugs, and may be suitably used according to a conventional method, but is not limited thereto. The amount of the active ingredient to be mixed can be suitably determined according to the intended use (prevention, health, or improvement). The composition of the quasi-drug composition of the present invention is not particularly limited, but it is preferable to use a disinfectant, a shower foam, a goggle, a wet tissue, a detergent soap, a spray, a hand wash, an ointment, a coating agent, a cream agent, a cataplasma agent, A sanitary pad, a sanitary napkin, a condom, a band, a filter filler, a sanitary glove, a sanitary container, a sanitary bag or a mask, and the form that can be manufactured by the above example is not limited.
본 발명에 따른 약학 조성물, 건강식품 조성물, 사료 조성물, 동물의약품 조성물 또는 의약외품 조성물은 결핵균에 대한 항균활성 또는 항균효과를 가짐으로써 결핵의 치료, 개선 또는 예방용으로 유용하게 사용할 수 있으며 다양한 항균제의 용도로 이용될 수 있다.
The pharmaceutical composition, health food composition, feed composition, animal medicine composition or quasi-drug composition according to the present invention have antimicrobial activity or antibacterial effect against Mycobacterium tuberculosis, so that they can be effectively used for treatment, improvement or prevention of tuberculosis. . ≪ / RTI >
도 1은 10E,12Z 공액리놀레산을 액체배지 발색법(Resazurin microtiter assay)에 적용하여 결핵균에 대한 상기 시험물질의 각 농도별 항균활성을 측정한 결과를 나타낸다. 또한 "+" 는 결핵균에 대한 시험물질의 항균활성 또는 항균효과를 나타낸다.
도 2는 10E,12Z 공액리놀레산을 MGIT SYSTEM 측정법에 적용하여 결핵균에 대한 상기 시험물질의 각 농도별 항균효과를 측정한 결과를 나타낸다. Fig. 1 shows the result of measuring the antibacterial activity of each test substance against the Mycobacterium tuberculosis by applying 10E, 12Z conjugated linoleic acid to a Resazurin microtiter assay. "+" Indicates the antibacterial activity or antibacterial effect of the test substance against Mycobacterium tuberculosis.
Fig. 2 shows the result of measuring the antibacterial effect of each test compound on the Mycobacterium tuberculosis by applying 10E, 12Z conjugated linoleic acid to MGIT SYSTEM measurement method.
이하, 본 발명을 실시 예를 통하여 보다 상세하게 설명한다. 그러나 이들 실시 예는 본 발명을 예시적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시 예에 한정되는 것은 아니다.
Hereinafter, the present invention will be described in more detail with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited to these examples.
: 재료: material
Middlebrook 7H9 Broth(Difco), Middlebrook ADC Enrichment supplement 용액, MGIT 7 Tube 및 MGIT Supplement(OADC)는 벡톤 디킨슨(Becton Dickinson, 미국)에서 구입하였다. Rifampicin, Isoniazid, Tween 80, Glycerol, DMSO(Dimethyl Sulfoxide), 에탄올 및 Resazurin sodium salt powder는 시그마 알드리치 주식회사(Sigma-Aldrich Co. LLC, St. Louis, MO, 미국)에서 구입하였다.
Middlebrook 7H9 Broth (Difco), Middlebrook ADC Enrichment supplement solution, MGIT 7 Tube and MGIT Supplement (OADC) were purchased from Becton Dickinson (USA). Rifampicin, Isoniazid, Tween 80, Glycerol, DMSO (Dimethyl Sulfoxide), ethanol and Resazurin sodium salt powder were purchased from Sigma-Aldrich Co. LLC, St. Louis, Mo., USA.
: 결핵균의 배양 : Culture of Mycobacterium tuberculosis
결핵균(H37Rv, ATCC 27294)을 배양하기 위한 액체배지는 0.05% Tween 80 또는 0.2% glycerol, 10% OADC Enrichment를 포함하는 Middlebrook 7H9 배지를 사용하였으며 배지에 결핵균 접종 후 shaking하면서 37 에서 배양하였다. 결핵균의 배양은 600 nm에서 흡광도가 0.09 (McFarland turbidity standard 0.5)에 도달할 때까지 배양하였으며 배양 후 소량씩 분주하여 -80에 냉동보관하고 실험 시 녹여서 사용하였다.
Middlebrook 7H9 medium containing 0.05% Tween 80 or 0.2% glycerol and 10% OADC Enrichment was used as a liquid culture medium for culturing Mycobacterium tuberculosis (H37Rv, ATCC 27294). The cultures were incubated at 600 nm until the absorbance reached 0.09 (McFarland turbidity standard 0.5). After incubation, the cells were subcultured and stored at -80 ° C for freezing.
: 액체배지 발색법(Resazurin microtiter assay;REMA)에 의한 항균활성 측정 : Measurement of antimicrobial activity by Resazurin microtiter assay (REMA)
결핵균(Mycobacterium tuberculosis)에 대한 10E,12Z 공액리놀레산의 항균활성을 확인하기 위하여 단일물질을 에탄올로 용해하고 배지를 사용하여 농도별로 제조하여 사용하였다. 약물대조군 약물로 Rifampicin과 Isoniazid를 사용하였으며 DMSO(Dimethyl Sulfoxide)로 용해한 후 사용하였다. To confirm the antimicrobial activity of 10E, 12Z conjugated linoleic acid against Mycobacterium tuberculosis, a single substance was dissolved in ethanol and prepared by concentration using the medium. Rifampicin and Isoniazid were used as drug control drugs and dissolved in DMSO (Dimethyl Sulfoxide).
결핵균은 배양 후 2×106 CFU/㎖이 되도록 Middlebrook 7H9 배지로 희석하여 준비하였고 결핵균 배양 배지에 상기 제조한 시험물질을 25 ㎍/㎖, 50 ㎍/㎖, 100 ㎍/㎖, 200 ㎍/㎖ 농도가 되도록 투여하고 96well plate에 200㎕씩 첨가하였다. 음성대조군, 양성대조군과 함께 약물대조군인 Rifampicin과 Isoniazid는 결핵균 배양배지에 각각 10 ㎍/㎖농도가 되도록 투여하고 96well plate에 200㎕씩 첨가한 후 96well plate를 비닐로 보양하여 37℃에서 5일간 배양하였다. 배양 5일 후 발색용액(0.02% resazurin solution) 30㎕를 96well plate의 각 well에 첨가하고 호일로 보양한 후 37℃에서 36시간 동안 재배양 하였다. 36시간 후 각 시험약물의 항균활성을 음성대조군, 양성대조군 및 약물대조군과 비교 측정하였다. 즉, 음성대조군으로 결핵균과 약물이 접종되지 않은 배지만 배양된 그룹(Blank Control Group), 양성대조군으로 결핵균만 배양된 그룹(Untreated Positive Growth Control Group), 약물대조군으로 결핵균에 Rifampicin과 Isoniazid가 투여된 그룹(Drug-Treated Group)을 함께 배양하여 결핵균에 대한 시험물질의 항균활성을 비교 측정하였다. After culturing, the Mycobacterium tuberculosis was diluted with Middlebrook 7H9 medium to a concentration of 2 × 10 6 CFU / ml. The test substances were added to the Mycobacterium tuberculosis culture medium at a concentration of 25 μg / ml, 50 μg / ml, 100 μg / And added to each well of a 96-well plate. Rifampicin and Isoniazid were added to the culture medium of Mycobacterium tuberculosis at a concentration of 10 μg / ml, and then added to a 96-well plate. The 96-well plate was incubated at 37 ° C. for 5 days Respectively. Five days after the incubation, 30 μl of a coloring solution (0.02% resazurin solution) was added to each well of a 96-well plate, and the plate was incubated at 37 ° C. for 36 hours. After 36 hours, the antimicrobial activity of each test drug was compared with the negative control, the positive control and the drug control. (Blank Control Group), a positive control group (Untreated Positive Growth Control Group), a drug control group, which received Rifampicin and Isoniazid as a drug control group (Drug-Treated Group) were cultured together to measure the antibacterial activity of test substances against Mycobacterium tuberculosis.
그 결과, 액체배지 발색법에 의한 항균활성 실험을 통하여 시험물질인 10E,12Z 공액리놀레산은 결핵균에 매우 높은 항균활성을 나타내고 있음이 확인되었다(도 1). 이러한 결과는 10E,12Z 공액리놀레산이 결핵균에 대한 항균용 조성물로서 이용 가능함을 의미한다.
As a result, it was confirmed that the
: MGIT SYSTEM 측정법에 의한 항균효과 측정 : Measurement of antimicrobial effect by MGIT SYSTEM measurement method
결핵균에 대한 10E,12Z 공액리놀레산의 항균효과를 확인하기 위하여 상기 시험물질을 에탄올에 녹인 후 배지를 사용하여 농도별로 제조한 후 MGIT SYSTEM 측정법에 적용하여 결핵균에 대한 시험물질의 항균효과를 측정하였다. 즉, 양성대조군으로 결핵균만 배양된 그룹(Untreated Positive Growth Control)과 함께 약물대조군으로 Rifampicin과 Isoniazid가 각각 10 ㎍/㎖ 농도로 투여된 그룹(Drug-treated Control)을 함께 배양하여 결핵균에 대한 시험물질의 항결핵균 효과를 비교 측정하였으며 4주동안 배양, 측정되었다. In order to confirm the antimicrobial effect of 10E, 12Z conjugated linoleic acid on the Mycobacterium tuberculosis, the above test substance was dissolved in ethanol, prepared by concentration using the medium, and then applied to the MGIT SYSTEM measurement method to measure the antibacterial effect of the test substance on the Mycobacterium tuberculosis. In other words, the group (Untreated Positive Growth Control), which was only a positive control group, was cultured with Drug-treated Control (Rifampicin and Isoniazid) at 10 ㎍ / ㎖ The effect of anti - tubercle bacilli was measured and cultured for 4 weeks.
MGIT SYSTEM 측정법의 과정은 다음과 같다. 먼저 MGIT 7㎖ Tube(Becton Dickinson, USA)에 MGIT Supplement 용액(OADC, Becton Dickinson, USA) 0.8㎖, 결핵균 용액 0.1㎖와 시험물질 용액을 농도별로 첨가한 후 Middlebrook 7H9 배지를 사용하여 최종부피를 8㎖로 조정하고 혼합 후 Tube를 BACTEC MGIT 960 SYSTEM(Becton Dickinson, USA)에 장착하여 배양 측정한다. 결핵균은 Middlebrook 7H9 배지로 배양하여 총 부피 8㎖를 기준으로 1,200,000 CFU/㎖에 맞게 조정하여 Tube에 첨가되었다. MGIT SYSTEM은 배양기기내에 결핵균의 성장과 분열증식을 자동으로 측정하는 형광측정 센서가 장착되어 있으며 결핵균의 분열증식 및 성장 유무에 따라 결핵균 또는 결핵균의 감수성이 자동으로 감지되어 측정된다. The procedure of the MGIT SYSTEM measurement method is as follows. First, 0.8 ml of MGIT supplement solution (OADC, Becton Dickinson, USA), 0.1 ml of Mycobacterium tuberculosis solution and test substance solution were added to each MGIT 7 ml tube (Becton Dickinson, USA) Ml. After mixing, the tube is mounted on a BACTEC MGIT 960 system (Becton Dickinson, USA) and cultured. Mycobacterium tuberculosis was cultured in Middlebrook 7H9 medium and added to the tube adjusted to 1,200,000 CFU / ml based on a total volume of 8 ml. The MGIT system is equipped with a fluorescence measurement sensor that automatically measures the growth and proliferation of Mycobacterium tuberculosis in the incubator and the susceptibility of Mycobacterium tuberculosis or Mycobacterium tuberculosis is automatically detected and measured according to the proliferation and growth of Mycobacterium tuberculosis.
그 결과, MGIT SYSTEM 측정법을 통하여 시험물질인 10E,12Z 공액리놀레산은 결핵균에 농도-의존적으로 매우 높은 항균활성 또는 항균효과를 나타내고 있음이 확인되었다(도 2). 이러한 결과들은 10E,12Z 공액리놀레산이 결핵균에 대한 항균용 조성물로서 이용 가능함을 의미한다. As a result, it was confirmed through the MGIT SYSTEM measurement method that 10E, 12Z conjugated linoleic acid, which is a test substance, showed a very high antimicrobial activity or antimicrobial effect in a dose-dependent manner to the Mycobacterium tuberculosis (Fig. 2). These results indicate that 10E, 12Z conjugated linoleic acid can be used as an antimicrobial composition against Mycobacterium tuberculosis.
상술한 바와 같이 본 발명에 대한 상기 시험물질의 측정결과는 도 1 내지 도 2에 나타내었다.
As described above, the measurement results of the test materials of the present invention are shown in FIGS. 1 and 2. FIG.
본 발명에 따른 약학 조성물, 건강식품 조성물, 사료 조성물, 동물의약품 조성물 및 의약외품 조성물은 결핵균에 대한 항균활성 또는 항균효과를 가짐으로써 결핵의 치료, 개선 또는 예방용으로 사용할 수 있으며 다양한 항균제의 용도로 이용될 수 있다. 본 발명에 따른 조성물은 산업적으로 제약업, 식품업, 농수산업 및 제조업의 산업분야에 유용하게 사용될 수 있다.
The pharmaceutical composition, health food composition, feed composition, animal medicine composition and quasi-drug composition according to the present invention have antimicrobial activity or antibacterial effect against Mycobacterium tuberculosis, so that they can be used for treatment, improvement or prevention of tuberculosis and used for various antibacterial agents . The composition according to the present invention can be industrially usefully used in industrial fields such as pharmaceutical industry, food industry, agriculture industry and manufacturing industry.
도 1에서 "+"는 결핵균에 대한 시험물질의 항균활성 또는 항균효과를 나타낸다. In Fig. 1, "+" indicates the antibacterial activity or antibacterial effect of the test substance against Mycobacterium tuberculosis.
Claims (6)
10E, 12Z conjugated linoleic acid as an active ingredient.
The composition of claim 1, wherein the composition has an antibacterial activity against mycobacterium tuberculosis.
10E, 12Z A health food composition for preventing or ameliorating tuberculosis containing conjugated linoleic acid as an active ingredient.
10E, 12Z A feed composition for preventing or improving tuberculosis containing conjugated linoleic acid as an active ingredient.
10E, 12Z Animal pharmaceutical composition for the treatment, improvement or prevention of tuberculosis containing conjugated linoleic acid as an active ingredient.
10E, 12Z A quasi-drug composition for preventing or improving tuberculosis containing conjugated linoleic acid as an active ingredient.
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