KR20170100236A - Health functional food comprising mixture of DNA fragments for preventing or improving osteoarthritis - Google Patents
Health functional food comprising mixture of DNA fragments for preventing or improving osteoarthritis Download PDFInfo
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- KR20170100236A KR20170100236A KR1020160022533A KR20160022533A KR20170100236A KR 20170100236 A KR20170100236 A KR 20170100236A KR 1020160022533 A KR1020160022533 A KR 1020160022533A KR 20160022533 A KR20160022533 A KR 20160022533A KR 20170100236 A KR20170100236 A KR 20170100236A
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
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- A—HUMAN NECESSITIES
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- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention
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Abstract
Description
본 발명은 DNA 단편 혼합물을 포함하는 관절염의 예방 또는 개선용 건강기능식품에 관한 것이다.The present invention relates to a health functional food for preventing or ameliorating arthritis comprising a DNA fragment mixture.
관절염은 인류가 앓고 있는 질환 중 유병률 1위의 다발성 질환으로, 특히, 퇴행성 관절염의 경우에는 65세 이상 인구의 70~80%가 지니고 있으며, 75세 이상의 인구에서는 거의 100%가 지니는 대표적인 노인성 질환으로 알려져 있다. 또한, 평균수명 연장과 함께 사회활동 기간이 늘어남에 따라, 퇴행성 관절염은 노년 생활의 편안한 생활 영위를 위한 질적 측면에 있어서 시급히 극복해야 할 노인성 질환 중에서도 가장 중요한 위치를 차지하고 있다.Arthritis is a multifactorial disease with the highest prevalence among the diseases suffered by mankind. In the case of degenerative arthritis, it has 70 ~ 80% of the population aged 65 and over. It is a representative geriatric disease with almost 100% It is known. In addition, as the life expectancy and the duration of social activities increase, degenerative arthritis is the most important of the geriatric diseases that need to be overcome urgently in terms of qualitative aspect for the elderly living comfort.
관절염은 신체의 근 골격 및 결합 조직에 염증성 변화가 생겨 근 골격계에 전신 증상이 나타나는 것을 총칭하는 병명이다. 또한, 관절염은 관절, 뼈, 연골 조직 또는 척수에 영향을 주어, 종종 영구적인 조직 손상, 기형, 퇴화 및 장애를 일으키는 만성염증을 특징으로 하며(Hofbause, L. C. et al., 2001), 퇴행성 관절염(골관절염), 류마티스 관절염, 척추관절병증, 강직성 척추염, 건선관절염, 통풍, 세균성 관절염, 소아기 류마티스관절염, 루푸스, 경피증, 다발성 경화증, 섬유근통, 다발성 근염, 피부근염, 라임 관절염, 유착 관절낭염, 오십견 등이 포함된다. Arthritis is a disease that collectively refers to the appearance of systemic symptoms in the skeletal system due to inflammatory changes in the body's skeleton and connective tissue. Arthritis also affects joints, bones, cartilage tissues or spinal cord, often characterized by chronic inflammation leading to permanent tissue damage, deformity, degeneration and disability (Hofbause, LC et al., 2001), degenerative arthritis Osteoarthritis), rheumatoid arthritis, spondyloarthropathies, ankylosing spondylitis, psoriatic arthritis, gout, bacterial arthritis, childhood rheumatoid arthritis, lupus, scleroderma, multiple sclerosis, fibromyalgia, multiple myositis, dermatomyositis, Lyme arthritis, .
관절염 질환 중에서도 가장 흔히 나타나는 퇴행성 관절염은, 퇴행성 관절 질환, 골관절염이라고도 불리며, 관절을 보호하고 있는 연골의 점진적인 손상이나 퇴행성 변화로 인해 관절을 이루는 뼈와 인대 등에 손상이 일어나 관절 주변의 활액막에 염증이 생기면서 통증과 변형이 발생하는 질환으로, 노령, 비만, 관절의 외상이나, 고관절 이형성증, 관절염의 과거력, 일부 특수직업군, 유전적 요인 등에 의하여, 무릎과 발목, 손가락, 팔꿈치 고관절 등과 같은 관절에서의 연골 조직이 점진적으로 퇴행되고 파괴되는 과정으로 진행된다. Degenerative arthritis, which is the most common arthritic disease, is also called degenerative joint disease and osteoarthritis. The progressive damage or degeneracy of the cartilage that protects the joint causes damage to the bones and ligaments of the joint, resulting in inflammation of the synovium around the joint It is a disease that causes pain and deformation. It is caused by the trauma of the elderly, obesity, joint, hip dysplasia, history of arthritis, some special occupation group, genetic factors, Progresses to progressive progression of cartilage tissue destruction.
또한, 특별한 기질적 원인 없이 나이, 성별, 유전적 요소, 비만, 특정 관절 부위 등의 요인에 따라 발생하는 일차성 또는 특발성 관절염과 관절 연골에 손상을 줄 수 있는 외상, 질병 및 기형 등이 원인이 되어 발생하는 이차성 또는 속발(성) 관절염으로 분류하며, 노령 인구의 증가에 따라 그 발병확률도 증가하는 추세이고, 나이가 많아질수록 여성에게서 더 많이 나타나는 경향이 있다. In addition, it may be caused by primary or idiopathic arthritis caused by factors such as age, gender, genetic factors, obesity, specific joint area, trauma, disease and deformity that may damage articular cartilage And the probability of onset is increasing with the increase of the elderly population. The older the age, the more tendency to appear in women.
퇴행성 관절염을 예방하거나 치료할 수 있는 확실한 약물은 개발되어 있지 않아, 진통 및 소염 작용을 가진 약품들이 현재 사용되고 있으며, 비스테로이드성 소염제가 대표적인 약제로 가장 많이 사용되고 있으나 장기 투여시 소화기 계통 및 응고 기전의 부작용이 있을 수 있다. 또한, 소화기 계통의 부작용을 줄여주는 새로운 기전의 비스테로이드성 소염제가 개발되었으나 이 역시 심혈관 계통 부작용의 가능성이 있어 사용 시 주의가 필요하다. 상기와 같이 비수술적 치료 방법에도 불구하고 더 이상 증상의 호전이 없으며 관절의 변화가 계속 진행되어 일상생활에 지장이 극심한 경우에는 수술적 치료 방법을 실시하게 되는데, 수술적 치료 중, 관절경에 의한 수술 방법은 비교적 간단한 수술로 증상 완화를 기대할 수 있으나 그 효과의 지속 여부가 일정하지 않다. 또한, 퇴행성 관절염의 대표적인 수술 방법인 인공 관절 치환술의 경우 효과적인 통증의 경감을 얻을 수 있고 변형된 관절이 교정되는 효과가 있으나, 인공 관절의 수명이 제한적이기 때문에 향후 재치환술을 필요로 할 수 있고, 수술 과정에 있어 출혈이나 감염 등의 합병증이 발생할 수 있어 주의가 요구된다.Although no definitive drug has been developed to prevent or treat degenerative arthritis, analgesic and anti-inflammatory drugs are currently in use and non-steroidal anti-inflammatory drugs are the most commonly used drugs. However, side effects of the digestive system and coagulation mechanism This can be. In addition, a new nonsteroidal antiinflammatory agent has been developed to reduce the side effects of the digestive system, but this may also be a side effect of the cardiovascular system. In addition to the above mentioned non-surgical treatment methods, there is no improvement in the symptoms. If the joint changes continuously and the daily life is disturbed, surgical treatment is performed. In the surgical treatment, Surgery is a relatively simple procedure that can be expected to relieve symptoms, but the effect is not constant. In the case of artificial joint arthroplasty, which is a representative surgical procedure for degenerative arthritis, effective pain relief can be obtained and deformed joints can be corrected. However, since the life of artificial joints is limited, In the surgical procedure, complications such as hemorrhage or infection may occur, requiring caution.
한편, 퇴행성 관절염에 대해 식품 섭취를 통한 예방도 가능하다는 것이 알려지면서 건강기능식품 시장이 확대되고 있다. 특히, 글루코사민과 콘드로이틴이 관절건강에 효과가 있는 제품으로 알려지면서, 이와 관련된 건강기능식품의 대다수 시장을 점유하고 있었으나, 최근에는 상기 물질이 퇴행성 관절염의 예방 효과나 이외 질환 치료 효과에 대한 근거가 없는 것으로 밝혀져(Clegg, D. O. et al., 2006), 관절 기능 보호와 향상을 위해 글루코사민과 콘드로이틴을 복용해 온 퇴행성 관절염 환자들에게 새로운 관절염의 예방 또는 개선용 건강기능식품을 제공하기 위한 원료의 개발이 시급한 실정이다. Meanwhile, it is known that degenerative arthritis can be prevented through food intake, and the market for health functional foods is expanding. In particular, glucosamine and chondroitin are known to be effective products for joint health and have occupied the majority of the market for health functional foods. Recently, however, there has been no evidence for the effectiveness of these substances in preventing or treating degenerative arthritis (Clegg, DO et al., 2006), the development of raw materials to provide health functional foods for the prevention or amelioration of new arthritis in patients with degenerative arthritis who have taken glucosamine and chondroitin to protect and improve joint function It is urgent.
관절염과 관련된 선행문헌으로서, 한국공개특허 제10-2008-0079038호에는 뉴클레오프로테인, DNA 및 RNA로부터 선택된 적어도 1종을 유효성분으로 함유하는 관절 류머티즘 예방을 위한 영양보조식품을 개시하고 있으며, 한국공개특허 제10-2013-0044195호에는 어류 정소로부터 분리된 DNA 단편 혼합물을 포함하는 연골 재생용 조성물을 개시하고 있고, 국제공개특허 제2010-049898호에는 어류의 정액으로부터 분리된 폴리데옥시리보뉴클레오티드를 포함하는 관절염 치료용 주사제 조성물에 대해 개시되었으나, 연어과에서 추출한 DNA 단편 혼합물 중, 특정 분자량을 선택하여 혼합한 혼합물을 함유하는 건강기능식품에 대해서는 아직까지 개시된 바 없다.As a prior art related to arthritis, Korean Patent Laid-Open No. 10-2008-0079038 discloses a nutritional supplement for preventing rheumatoid arthritis containing at least one selected from nucleoprotein, DNA and RNA as an active ingredient, Patent Publication No. 10-2013-0044195 discloses a composition for regenerating cartilage comprising a DNA fragment mixture separated from a fish testimony. International Patent Publication No. 2010-049898 discloses a composition comprising a polydeoxyribonucleotide , There has not yet been disclosed a health functional food containing a mixture of a DNA fragment mixture extracted from Salmonellae and a mixture of selected molecular weights.
본 발명의 목적은 DNA 단편 혼합물을 포함하는 관절염의 예방 또는 개선용 건강기능식품을 제공하는 데 있으며, 특히, 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물로 구성된 DNA 단편 혼합물을 포함하는 관절염의 예방 또는 개선용 건강기능식품을 제공하는 데 있다. It is an object of the present invention to provide a health functional food for preventing or ameliorating arthritis comprising a DNA fragment mixture, and more particularly to a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa, Or less of a DNA fragment mixture comprising a DNA fragment mixture consisting of a DNA fragment mixture consisting of a DNA fragment mixture consisting of a DNA fragment mixture.
본 발명은 DNA 단편 혼합물을 포함하는 관절염의 예방 또는 개선용 건강기능식품에 관한 것이다. 상기 DNA 단편 혼합물은 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물이 혼합될 수 있으며, 상기 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물은 1~2:1~2:1~2의 중량비로 혼합될 수 있다. The present invention relates to a health functional food for preventing or ameliorating arthritis comprising a DNA fragment mixture. The DNA fragment mixture may be a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less, A DNA fragment mixture having a molecular weight of less than 2500 kDa and a DNA fragment having a molecular weight of 2500 kDa or more and 5000 kDa or less may be mixed at a weight ratio of 1: 2: 1 to 2: 1 to 2.
상기 DNA 단편 혼합물은 어류의 정액 또는 정소에서 분리하였고, 상기 어류는 연어과의 어류인 것을 특징으로 한다.The DNA fragment mixture is separated from the semen of a fish or a testis, and the fish is a fish of salmonid.
또한, 상기 관절염은 퇴행성 관절염(osteoarthritis)인 것을 특징으로 한다.In addition, the arthritis is characterized by degenerative arthritis (osteoarthritis).
이하 본 발명을 상세하게 설명한다.Hereinafter, the present invention will be described in detail.
본 발명은 DNA 단편 혼합물을 포함하는 관절염의 예방 또는 개선용 건강기능식품에 관한 것으로, 상기 DNA 단편 혼합물은 인산, 4종류의 염기, 데옥시리보오스로 이루어진 생체고분자에 해당하는 DNA가 어류 정소로부터 추출, 정제된 후 그 분자량이 저감된 단편 형태의 상태로 혼합되어 존재하는 것을 지칭하며, 백색 또는 미황색의 결정형 파우더로서 물과 알칼리에 조금 녹고, 알콜에 매우 녹기 어려우며, 에테르와 아세톤에 거의 녹지 않는 특성을 갖는다. The present invention relates to a health functional food for preventing or ameliorating arthritis, which comprises a DNA fragment mixture, wherein the DNA fragment mixture is obtained by extracting DNA corresponding to a biopolymer composed of phosphoric acid, four kinds of bases and deoxyribose from fish testis Refers to a white or yellowish crystalline powder which is slightly soluble in water and alkali and is hardly soluble in alcohol and hardly soluble in ether and acetone Respectively.
상기 DNA 단편 혼합물은 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물이 혼합될 수 있으며, 상기 제시된 분자량의 범위를 벗어나 5kDa미만의 분자량을 포함하는 DNA 단편 혼합물의 경우에는 관절염의 예방 또는 개선 효과가 충분히 나타나기 전에 소화가 되어 효과의 지속성이 떨어지므로 사용하는데 바람직하지 않고, 5000kDa을 초과하는 분자량을 포함하는 DNA 단편 혼합물의 경우에는 분리, 정제 또는 여과 등의 효율이 낮아 사용하는데 바람직하지 않다.The DNA fragment mixture may be a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa, or a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less, and having a molecular weight of less than 5 kDa Is not preferable for use because it is extinguished before the prevention or improvement effect of arthritis appears sufficiently, and the persistence of the effect becomes poor. In the case of a DNA fragment mixture containing a molecular weight exceeding 5000 kDa, The filtration efficiency is low and it is not preferable for use.
본 발명에 따르면, 상기 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물은 1~2:1~2:1~2의 중량비로 혼합될 수 있다. 관절염의 예방 또는 개선 효과를 위한 경구 투여시, 상기 혼합범위를 벗어나 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물이, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물과 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물에 비해 상대적으로 더 많은 중량비로 포함된 경우에는, 속효성 효과는 빠른 시간 내에 나타나지만 지속성이 유지되는 효과가 낮아 바람직하지 않다. 또한, 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물이, 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물과 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물에 비해 상대적으로 더 많은 중량비로 포함된 경우에는, 지속성 효과는 유지되나 속효성 효과가 빠른 시간 내에 나타나지 않아 바람직하지 않다. 또한, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물이, 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물과 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물에 비해 상대적으로 더 많은 중량비로 포함된 경우에는, 속효성이 비교적 빠른 시간 내에 나타나며 지속성 효과도 유지되지만, 본 발명에서 제시하는 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물을 1~2:1~2:1~2의 중량비로 혼합하여 경구 투여하는 것이, 속효성 효과 및 지속성 효과가 가장 우수하다.According to the present invention, a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less can be mixed at a weight ratio of 1: 2: 1 to 2: have. A DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa out of the above mixing range is relatively more than a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less at the time of oral administration for prevention or amelioration of arthritis When included in a weight ratio, the quick-acting effect appears within a short period of time, but is not preferable because the effect of maintaining persistence is low. Further, when the DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less is contained in a relatively larger weight ratio than a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa and a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa, the persistence effect is maintained, It is not desirable because it does not appear in a short time. Also, when a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa is contained at a relatively higher weight ratio than a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less, A DNA fragment mixture having a molecular weight of more than 50 kDa but less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less in a weight ratio of 1: 2: 1 to 2: 1 to 2 And the oral administration is most effective in the fast-acting effect and the persistence effect.
본 발명의 상기 DNA 단편 혼합물은 어류의 정액 또는 정소에서 분리하였고, 상기 어류는 연어과의 어류인 것이 바람직하며, 상기 관절염은 퇴행성 관절염인 것을 특징으로 한다.The DNA fragment mixture of the present invention is isolated from the semen of a fish or a testis, and the fish is preferably a fish of salmonidia, and the arthritis is degenerative arthritis.
본 발명에 따른 상기 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물로 구성된 DNA 단편 혼합물은 경구 투여가 가능하며, 경구 투여를 위한 사용형태는 특별히 한정되지 않고, 건강기능식품법 상으로 허용하는 부형제와 섞어 산제, 정제, 캡슐제, 환제, 과립제, 분말, 액제 등의 형태를 포함할 수 있다.The DNA fragment mixture according to the present invention comprising a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa, or a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less can be administered orally, And may be in the form of powders, tablets, capsules, pills, granules, powders, liquid preparations mixed with excipients allowed in the health functional food method.
또한, 상기 DNA 단편 혼합물은 식품학적으로 허용 가능한 식품보조 첨가제가 첨가될 수 있는데, 예를 들어, 조미료, 착향료, 감미료, 산미료, 보존제, 첨가제 및 강화제 등이 포함된다. 본 발명의 혼합물을 첨가할 수 있는 식품으로는, 예를 들어, 육류, 소세지, 빵, 쵸코렛, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 건강기능식품을 모두 포함한다. 본 발명의 건강음료 조성물은 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 포함할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드 및 덱스트린, 사이클로덱스트린과 같은 폴리사카라이드, 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 감미제로서는 타우마틴, 스테비아 추출물과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100㎖당 일반적으로 약 0.01~0.04g, 바람직하게는 약 0.02~0.03g이다. 상기 외에 본 발명의 건강기능식품은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산음료에 사용되는 탄산화제 등을 포함할 수 있다. 그밖에 본 발명의 건강기능식품은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 포함할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 크게 중요하진 않지만 본 발명의 건강기능식품 100 중량부 당 0.01~0.1 중량부의 범위에서 선택되는 것이 일반적이다.In addition, the above DNA fragment mixture may be supplemented with a food-acceptable food-aid additive, for example, a seasoning, an flavoring agent, a sweetener, an acidulant, a preservative, an additive and a reinforcing agent. Foods to which the mixture of the present invention can be added include food products such as meat, sausage, bread, chocolate, candy, snack, confectionery, pizza, ramen, other noodles, gums, ice cream, Drinks, tea, drinks, alcoholic beverages, and vitamin complexes, all of which include health functional foods in a conventional sense. The health beverage composition of the present invention may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, and polysaccharides such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol and erythritol. Examples of sweeteners include natural sweeteners such as tau martin and stevia extract, synthetic sweeteners such as saccharin and aspartame, and the like. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 ml of the composition of the present invention. In addition to the above, the health functional food 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, Alcohols, carbonating agents used in carbonated drinks, and the like. In addition, the health functional food of the present invention may include natural fruit juice, fruit juice drink, and pulp for producing vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health functional food of the present invention.
본 발명은 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물로 구성된 DNA 단편 혼합물을 포함하는 관절염의 예방 또는 개선용 건강기능식품에 관한 것으로서, 상기 DNA 단편 혼합물은 경구 투여시, 퇴행성 관절염의 예방지표로 알려진 염증성 사이토카인의 생성을 유의하게 억제하고, 속효성 및 지속성이 유지되는 효과가 우수하여 관절염의 예방 또는 개선용 건강기능식품으로 유용하게 사용될 수 있다.The present invention relates to a health functional food for preventing or ameliorating arthritis, which comprises a DNA fragment mixture of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less, The DNA fragment mixture significantly inhibits the production of inflammatory cytokine, which is known as a preventive index of degenerative arthritis, when administered orally, and is excellent in the effect of maintaining the rapidity and persistence, and thus can be useful as a health functional food for preventing or improving arthritis have.
도 1은 퇴행성 관절염이 유발된 동물모델에서의 본 발명의 실시예 4-1에 대한 IL-1β, IL-6, TNF-α, COX-2, iNOS, BCL-2 및 BAX의 단백질 발현을 측정한 그래프이다. 1 shows the expression of IL-1β, IL-6, TNF-α, COX-2, iNOS, BCL-2 and BAX in the animal model induced by degenerative arthritis according to Example 4-1 of the present invention It is a graph.
이하 본 발명의 바람직한 실시예를 상세히 설명하기로 한다. 그러나 본 발명은 여기서 설명되는 실시예에 한정되지 않고 다른 형태로 구체화될 수도 있다. 오히려, 여기서 소개되는 내용이 철저하고 완전해지고, 당업자에게 본 발명의 사상을 충분히 전달하기 위해 제공하는 것이다.Hereinafter, preferred embodiments of the present invention will be described in detail. However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Rather, the intention is to provide an exhaustive, complete, and complete disclosure of the principles of the invention to those skilled in the art.
<실시예 1. 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물의 제조>≪ Example 1: Preparation of a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa >
(1) 정소 준비 공정(1) Test preparation process
냉동된 정소 10g을 해동한 다음 탈혈 및 이물질을 제거하고, 정제수 400㎖을 넣어준 후, 분쇄기로 분쇄하여 정소 분쇄액을 제조하였다.10 g of frozen testimony was thawed, and then blood was removed and foreign matter was removed, and 400 ml of purified water was added thereto, followed by pulverization with a pulverizer to prepare a testis pulverized liquid.
(2) 단백질 및 지방 제거 공정(2) Protein and fat removal process
이 후, 정제수 500㎖에 200g의 염화나트륨을 녹여 염화나트륨 용액을 제조하였고, 상기 염화나트륨 용액 500㎖와 상기 정소 분쇄액을 혼합한 다음, 100℃에서 1시간 동안 교반하면서 반응시킨 후에 냉각하였다. 상기 냉각된 반응액에 30% 수산화나트륨(NaOH)을 첨가하여 pH 11.0이 되도록 한 후 이를 1시간 동안 교반시키고 여과하여 단백질과 지방이 제거된 여과액을 얻었다.Thereafter, sodium chloride solution was prepared by dissolving 200 g of sodium chloride in 500 ml of purified water. 500 ml of the sodium chloride solution and the testis solution were mixed and reacted with stirring at 100 ° C for 1 hour, followed by cooling. 30% sodium hydroxide (NaOH) was added to the cooled reaction solution to adjust the pH to 11.0, which was then stirred for 1 hour and filtered to obtain a filtrate from which protein and fat were removed.
(3) 분자량 저감 공정(3) Molecular weight reduction process
상기 여과액에, pH 7.0이 되도록 염산을 첨가하여 100℃에서 10분간 반응시킨 후, 이를 냉각한 뒤 다시 여과를 실시하여 DNA 단편 혼합물을 포함하는 여과액을 얻었다.Hydrochloric acid was added to the filtrate to adjust the pH to 7.0, and the mixture was reacted at 100 ° C for 10 minutes. After cooling, the filtrate was filtered to obtain a filtrate containing a DNA fragment mixture.
(4) 분자량 분획 공정(4) Molecular weight fractionation process
여과액에 30%(w/v) 수산화나트륨 수용액을 첨가하여 다시 pH 10.0이 되도록 한 후 겔여과를 통해 5kDa이상 50kDa미만의 분자량을 가지는 DNA 단편 혼합물의 분획액을 얻은 다음 35%(w/v) 염산을 사용하여 pH 7.0으로 조정한다.To the filtrate, a 30% (w / v) aqueous solution of sodium hydroxide was added to bring the pH to 10.0 again. Then, a fraction of the DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa was obtained through gel filtration. ) ≪ / RTI > hydrochloric acid.
(5) 침전 및 건조 공정(5) Precipitation and drying process
상기 분획액에 냉각된 에탄올(여과액 부피의 2배)을 첨가한 다음 10분간 교반하고 원심분리한 뒤, 침전된 침전물을 회수하여 이를 70% 에탄올 수용액으로 세척하였다. 진공 하에서 건조시켜 DNA 단편 혼합물을 수득하였으며, 분말화된 DNA 단편 혼합물의 균일성을 확보하기 위하여 1㎜ 체에 거르는 과정을 반복하여, 5kDa이상 50kDa미만의 분자량이 포함된 DNA 단편 혼합물을 얻었다.Cooled ethanol (twice the volume of the filtrate) was added to the fraction solution, stirred for 10 minutes, centrifuged, and the precipitated precipitate was recovered and washed with 70% aqueous ethanol solution. And dried under vacuum to obtain a DNA fragment mixture. In order to ensure the uniformity of the powdered DNA fragment mixture, the process of filtering through a 1 mm sieve was repeated to obtain a DNA fragment mixture containing a molecular weight of 5 kDa or more and less than 50 kDa.
<실시예 2. 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물의 제조>≪ Example 2: Preparation of a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa &
상기 실시예 1과 같이 제조하되 분자량 분획 공정에서 겔여과를 통해 50kDa이상 2500kDa미만의 분자량을 가지는 DNA 단편 혼합물의 분획액을 얻은 다음 침전 및 건조 공정을 실시하여, 50kDa이상 2500kDa미만의 분자량이 포함된 DNA 단편 혼합물을 얻었다.A fraction solution of a DNA fragment mixture having a molecular weight of at least 50 kDa and less than 2500 kDa was obtained through gel filtration in the molecular weight fractionation step and then subjected to a precipitation and drying step to obtain a fraction containing a molecular weight of less than 2500 kDa A DNA fragment mixture was obtained.
<< 실시예Example 3. 분자량 3. Molecular weight 2500kDa이상2500kDa or more 5000kDa이하인Less than 5000 kDa DNA 단편 혼합물의 제조> Preparation of DNA fragment mixtures >
상기 실시예 1과 같이 제조하되 분자량 분획 공정에서 겔여과를 통해 2500kDa이상 5000kDa이하의 분자량을 가지는 DNA 단편 혼합물의 분획액을 얻은 다음 침전 및 건조 공정을 실시하여, 2500kDa이상 5000kDa이하의 분자량이 포함된 DNA 단편 혼합물을 얻었다.A fraction solution of a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less was obtained through gel filtration in the molecular weight fractionation step and then subjected to precipitation and drying to obtain a fraction containing a molecular weight of 2500 kDa or more and 5000 kDa or less A DNA fragment mixture was obtained.
<실시예 4. 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물로 이루어진 DNA 단편 혼합물의 제조>Example 4: Preparation of a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa, and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less>
상기 실시예 1 내지 3에서 분리 획득한 각각의 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물을 하기 표 1의 조건으로 혼합하여 실시예 4-1 내지 4-7을 제조하였다.A DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less respectively obtained in Examples 1 to 3 were mixed under the conditions shown in Table 1, -1 to 4-7.
5kDa이상 50kDa미만(g)Molecular Weight
5 kDa or more and less than 50 kDa (g)
50kDa이상 2500kDa미만(g)Molecular Weight
50 kDa or more and less than 2500 kDa (g)
2500kDa이상 5000kDa이하(g)Molecular Weight
2500 kDa or more and 5000 kDa or less (g)
<< 비교예Comparative Example 1. 분자량 5 내지 1. Molecular weight of 5 - 5000kDa의5000 kDa DNA 단편 혼합물의 제조> Preparation of DNA fragment mixtures >
상기 실시예 1과 같이 제조하되 분자량 분획 공정에서 겔여과를 통해 5kDa 내지 5000kDa의 분자량을 가지는 DNA 단편 혼합물의 분획액을 얻은 다음 침전 및 건조 공정을 실시하여, 분자량 5kDa 내지 5000kDa의 분자량이 포함된 DNA 단편 혼합물을 얻었다.A fraction containing the DNA fragments having a molecular weight of 5 kDa to 5000 kDa was obtained through gel filtration in the molecular weight fractionation step and then subjected to a precipitation and drying process to obtain a DNA having a molecular weight of 5 kDa to 5000 kDa A fragment mixture was obtained.
<비교예 2. 비교대상 DNA 단편 혼합물의 제조>≪ Comparative Example 2: Preparation of a DNA fragment mixture for comparison >
상기 실시예 1 내지 3의 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물을 하기 표 2의 조건으로 혼합하여 비교예 2-1 내지 2-9를 제조하였다. The DNA fragment mixtures having a molecular weight of 5 kDa or more and less than 50 kDa, the DNA fragment mixtures having a molecular weight of 50 kDa or more and less than 2500 kDa and the DNA fragment mixtures having a molecular weight of 2500 kDa or more and 5000 kDa or less in the above Examples 1 to 3 were mixed under the conditions shown in Table 2, 9 was prepared.
5kDa이상 50kDa미만(g)Molecular Weight
5 kDa or more and less than 50 kDa (g)
50kDa이상 2500kDa미만(g)Molecular Weight
50 kDa or more and less than 2500 kDa (g)
2500kDa이상 5000kDa이하(g)Molecular Weight
2500 kDa or more and 5000 kDa or less (g)
<실험예 1. in vitro에서의 속효성 확인>Experimental Example 1. Identification of rapidity in vitro [
실험에 사용하기 위한 RAW 264.7세포는 한국세포주은행(서울, 한국)에서 구입하였다. 상기 RAW 264.7 세포를 6웰 플레이트에 3×105 cells/㎖가 되도록 분주하고 24시간 동안 배양한 후, 100㎍/㎖의 각 실험군 및, 1㎍/㎖의 LPS를 처리하였다. 24시간 동안 배양한 후 세포배양액을 수거하여 배양액에 함유된 TNF-α를 Milliplex map cytokine kit(Millipore Co., Bellerica, USA)로 측정하였고, 분석은 Milliplex analyst software(고마바이오텍, 서울, 한국)를 통해 이루어졌다(참고: 대한한의학회지 제34권 제3호(2013년 9월)).RAW 264.7 cells for use in the experiments were purchased from Korean Cell Line Bank (Seoul, Korea). The RAW 264.7 cells were seeded at 6 × 10 5 cells / ml in a 6-well plate, cultured for 24 hours, treated with 100 μg / ml of each experimental group and 1 μg / ml of LPS. After culturing for 24 hours, the cell culture medium was collected, and the TNF-α contained in the culture solution was measured with a Milliplex map cytokine kit (Millipore Co., Bellerica, USA) and analyzed using Milliplex analyst software (Goma Biotech, Seoul, Korea) (Note: The Korean Journal of Medicine 34, Issue 3 (September 2013)).
상기 표 3을 참고하면, 본 발명의 실시예 4-1을 처리한 경우는 대조군 또는 비교예 1 및 비교예 2-1 내지 2-3을 처리한 경우에 비해, 처리 1시간 후부터 TNF-α의 발현율이 우수하게 감소하는 것을 확인할 수 있어, 본 발명의 DNA 단편 혼합물은 관절염에 즉시적인 감소 효과를 나타내는 조성물임을 알 수 있다. The results are shown in Table 3. The results are shown in Table 3. The results are shown in Table 3. The results are shown in Table 3. The results are shown in Table 3. [ It was confirmed that the DNA fragment mixture of the present invention is a composition showing an immediate reduction effect on arthritis.
<< 실험예Experimental Example 2. 동물모델에서의 퇴행성 관절염 유발> 2. Degenerative arthritis induction in animal models>
무릎관절은 콜라겐(collagen), 프로테오글리칸(proteoglycan) 및 연골세포로 구성되어 있는데, 콜라겐분해효소(collagenase) 주입은 콜라겐(collagen)을 분해하여 퇴행성 관절염을 유발하는 방법이며, MIA(monosodium iodoacetate) 주입은 단백분해효소(matrix metalloproteinase)의 활성을 증가시키고 연골에서의 프로테오글리칸 합성을 억제하여 결과적으로 연골 세포의 괴사를 일으켜 비교적 단시간에 퇴행성 관절염을 유발하는 방법이다. 이에, 본 발명에서는 동물모델의 무릎 관절 내에 MIA를 주입하여 퇴행성 관절염이 유발된 동물모델을 얻었다. Knee joints consist of collagen, proteoglycan and cartilage cells. Collagenase injection is a method of decomposing collagen to cause degenerative arthritis. MIA (monosodium iodoacetate) injection It is a method to increase the activity of matrix metalloproteinase and to inhibit proteoglycan synthesis in cartilage, resulting in necrosis of cartilage cells and induce degenerative arthritis in a relatively short time. In the present invention, an animal model in which degenerative arthritis was induced by injection of MIA into the knee joint of an animal model was obtained.
이를 위해, 먼저 10주령 SD 랫트(Sprague-Dawley rat)를 에틸 에테르(ethyl ether)로 마취시킨 후, 오른쪽 무릎 주변을 제모하고 퇴행성 관절염 유발 물질인 MIA 1㎖를 인슐린 주사기로 오른쪽 무릎 관절 내에 50㎕(60㎎/㎖)씩 투여하였다. MIA의 희석은 생리식염수(0.9% saline)를 사용하였으며, 약물 주사 7일 후에 랫트의 관절 부위에 나타난 부종의 정도와 걷는 상태를 관찰하여 퇴행성 관절염이 성공적으로 유발된 것을 확인한 후 실험을 진행하였다.To this end, 10-week old SD rats (Sprague-Dawley rats) were anesthetized with ethyl ether, and the vicinity of the right knee was removed. 1 ml of MIA, a degenerative arthritis inducing substance, was injected into the right knee joint (60 mg / ml). MIA was diluted with physiological saline (0.9% saline) and after 7 days of drug injection, the degree of edema at the joint area of the rat and the condition of walking were observed to confirm the successful induction of degenerative arthritis.
<실험예 3. 퇴행성 관절염이 유발된 동물모델에서의 IL-1β, IL-6, TNF-α, COX-2, iNOS, BCL-2 및 BAX의 단백질 발현 양상 확인>EXPERIMENTAL EXAMPLE 3. Confirmation of Protein Expression Patterns of IL-1β, IL-6, TNF-α, COX-2, iNOS, BCL-2 and BAX in Animal Model Induced Degenerative Arthritis
DNA 단편 혼합물의 투여에 따른 연골세포의 염증발현을 확인하고자, 상기 실험예 2에서 퇴행성 관절염이 유발된 동물모델로부터 연골조직을 채취하여 염증반응의 마커인 IL-1β, IL-6, TNF-α, COX-2 및 iNOS, 연골세포사멸을 막는 단백질인 BCL-2, 세포사멸을 유도하는 단백질인 BAX의 단백질 발현 변화를 확인하였다. In order to confirm the expression of inflammation of chondrocytes by the administration of the DNA fragment mixture, cartilage tissue was collected from the animal model in which degenerative arthritis was induced in Experimental Example 2, and IL-1β, IL-6, TNF-α , COX-2 and iNOS, BCL-2, a protein that blocks chondrocyte apoptosis, and BAX, a protein that induces apoptosis.
먼저, 동일한 환경에서 사육한 60마리의 랫트를 각 그룹당 10마리씩 총 6그룹으로 랜덤하게 나눈 다음, 본 발명의 실시예 4-1의 DNA 단편 혼합물을 랫트의 무게(성인의 몸무게가 60kg일 때 섭취용량 1g을 기준으로 함)에 따라 1일 1회씩 4주간 경구투여하고, 같은 기간 동안 실시예 4-1 대신, 대조군(식염수 주입)을 투여하여 비교하였다. First, 60 rats raised in the same environment were randomly divided into 6 groups of 10 rats per each group. Then, the DNA fragment mixture of Example 4-1 of the present invention was weighed by the weight of the rats (when the adult weight was 60 kg, 1 g per dose) for 4 weeks once a day and compared with the control group (saline infusion) administered for the same period instead of Example 4-1.
이후, 실시예 4-1 또는 대조군이 처리된 랫트로부터 채취한 20㎍/㎖의 단백질을 SDS-PAGE(sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 8-10% Tris-glycine gel, Invitrogen co.,)에 로딩하여 1시간 동안 전기영동을 하여 단백질 분리를 진행한 후, semi trans blot을 이용하여 멤브레인에 단백질을 전이시켰다. 상기 단백질이 전이된 멤브레인을 탈지유가 함유된 TBS(tris-buffered saline)용액으로 블로킹한 뒤, IL-1β, IL-6, TNF-α, COX-2, iNOS, BCL-2 및 BAX의 각 1차 항체(1:1,000으로 TBS 용액에 희석)를 첨가하여 반응 시킨 후, TBS로 6회 세척하였다. 이후, 2차 항체(anti-rabbit antibody, 1:1,000으로 희석)를 더한 다음 상온에서 1시간 반응하고, TBS로 6회 세척 후, ECL 기질과 30~60초간 반응하여, LAS-3000(Imaging system)을 통해 밴드 및 밴드의 밀도를 측정하여 이를 도 1에 나타내었다.Then, 20 μg / ml of the protein collected from the rat in Example 4-1 or the control group was subjected to SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis, 8-10% Tris-glycine gel, Invitrogen co. After loading and electrophoresis for 1 hour, protein separation was performed, and protein was transferred to the membrane using semi trans blot. The protein-transduced membrane was blocked with a solution of TBS (tris-buffered saline) containing skimmed milk, and each 1 of IL-1β, IL-6, TNF-α, COX-2, iNOS, BCL- The secondary antibody (1: 1,000 diluted in TBS solution) was added and reacted, followed by 6 washes with TBS. After incubation for 1 hour at room temperature, the cells were washed 6 times with TBS, reacted with the ECL substrate for 30 to 60 seconds, and then lysed with LAS-3000 (Imaging system ) To measure the densities of bands and bands, and these are shown in Fig.
도 1을 참고하면, 퇴행성 관절염이 유발된 동물모델에 본 발명의 실시예 4-1을 처리하는 경우, 염증반응의 마커인 IL-1β, IL-6, TNF-α, COX-2, iNOS 및 BAX의 발현량은 감소시키며 연골세포사멸을 막는 단백질인 BCL-2의 발현량은 증가하는 결과를 나타내어, 본 발명의 DNA 단편 혼합물은 퇴행성 관절염의 예방 또는 개선용 건강기능식품으로 유용하게 사용될 수 있음을 알 수 있었다.IL-1β, IL-6, TNF-α, COX-2, iNOS and IL-1β, which are markers of the inflammatory response, are observed when treating Example 4-1 of the present invention in an animal model in which degenerative arthritis has been induced The expression level of BCL-2, which is a protein that inhibits chondrocyte apoptosis, is decreased and the DNA fragment mixture of the present invention is useful as a health functional food for preventing or improving degenerative arthritis. And it was found.
<실험예 4. 퇴행성 관절염이 유발된 동물모델에서의 지속성 확인>EXPERIMENTAL EXAMPLE 4. Confirmation of Persistence in Animal Model Induced Degenerative Arthritis [
퇴행성 관절염이 유발된 동물모델에서 본 발명의 DNA 단편 혼합물 투여에 따른 속효성 및 지속성을 확인하고자, 상기 실험예 2에서 퇴행성 관절염이 유도된 동물모델로부터 활액을 채취하여 염증반응의 마커로서 COX-2의 단백질 발현 변화를 확인하였다. In order to confirm the rapid efficacy and persistence of the DNA fragment mixture of the present invention in an animal model in which degenerative arthritis was induced, synovial fluid was collected from an animal model in which degenerative arthritis was induced in Experimental Example 2, and COX-2 Protein expression changes were confirmed.
이를 위해, 먼저, 동일한 환경에서 사육한 60마리의 랫트를 각 그룹당 10마리씩 총 6그룹으로 랜덤하게 나눈 다음, 본 발명의 실시예 4-1 내지 4-3의 DNA 단편 혼합물 각각을 랫트의 무게(성인의 몸무게가 60kg일 때 섭취용량 1g을 기준으로 함)에 따라 1일 1회씩 1주간 경구투여하고, 투여종료 후 1일, 4일, 7일 째의 COX-2 단백질 발현율을 확인하였다. 또한, 이때 같은 기간 동안 상기 실시예 대신, 비교예 1, 비교예 2-1 내지 2-3 및 대조군(식염수 주입) 각각을 투여하여 비교분석 하였다. For this purpose, first, 60 rats raised in the same environment were randomly divided into 6 groups of 10 rats per each group, and then each of the DNA fragment mixtures of Examples 4-1 to 4-3 of the present invention was weighed 1 g per day when the adult body weight was 60 kg), and COX-2 protein expression was observed on
상기 표 4를 참고하면, 본 발명의 실시예 4-1 내지 4-3의 DNA 단편 혼합물을 투여한 경우, 투여 종료 후에도 최대 7일까지 염증성 사이토카인 인자인 COX-2의 발현이 감소되어, 본 발명의 DNA 단편 혼합물의 항염증 활성이 지속적으로 유지되고 있음을 알 수 있었다. 즉, 퇴행성 관절염이 유발된 경우에 있어서, 본 발명의 DNA 단편 혼합물(분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물이 혼합)을 경구복용하면 퇴행성 관절염 개선 효과의 지속성을 확인할 수 있을 뿐만 아니라, 상기 실험예 1의 결과로부터 속효성 효과도 확인된 바, 지속성 및 속효성의 효과를 동시에 갖는 퇴행성 관절염의 예방 또는 개선용 건강기능식품으로 유용하게 사용될 수 있음을 알 수 있었다. Referring to Table 4, when the DNA fragment mixture of Examples 4-1 to 4-3 of the present invention was administered, expression of COX-2, an inflammatory cytokine factor, was decreased up to 7 days after the end of administration, It was found that the anti-inflammatory activity of the inventive DNA fragment mixture was maintained constantly. That is, when the degenerative arthritis is induced, the DNA fragment mixture of the present invention (a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less) It is possible to confirm the persistence of the degenerative arthritis improving effect as well as the quick-acting effect from the results of Experimental Example 1 as described above. As a result, it has been confirmed that the present invention is useful as a health functional food for preventing or improving degenerative arthritis .
<< 제제예Formulation example 1. One. 산제의Sanje 제조> Manufacturing>
본 발명의 실시예 4-1 2g, 유당 1g을 혼합하고 기밀포에 충진하여 산제를 제조하였다.2 g of Example 4-1 of the present invention and 1 g of lactose were mixed and filled in an airtight container to prepare a powder.
<제제예 2. 정제의 제조>≪ Preparation Example 2 > Preparation of Tablets >
본 발명의 실시예 4-1 100㎎, 옥수수전분 100㎎, 유당 100㎎ 및 스테아린산 마그네슘 2㎎을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조하였다.After mixing 100 mg of Example 4-1 of the present invention, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate, tablets were prepared by tableting according to a conventional preparation method of tablets.
<제제예 3. 캡슐제의 제조>≪ Formulation Example 3: Preparation of capsules >
본 발명의 실시예 4-1 100㎎, 옥수수전분 100㎎, 유당 100㎎ 및 스테아린산 마그네슘 2㎎을 혼합한 후 통상의 캡슐제 제조방법에 따라 상기의 성분을 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조하였다.After mixing 100 mg of Example 4-1 of the present invention, 100 mg of corn starch, 100 mg of lactose and 2 mg of magnesium stearate, the above components were mixed according to a conventional capsule preparation method and filled in a gelatin capsule to prepare a capsule .
<제제예 4. 환제의 제조>≪ Formulation Example 4: Preparation of pill &
본 발명의 실시예 4-1 90㎎, 찹쌀전분 5㎎ 및 정제수 5㎎ 및 흡습성을 저해하는 첨가제로서 덱스트린, 말토덱스트린, 옥수수전분, 미결정셀룰로오스(MCC)로 이루어진 군에서 1종 이상을 소량 혼합한 후, 통상의 방법에 따라 100㎎의 환제를 제조하였다.90 mg of Example 4-1 of the present invention, 5 mg of glutinous rice starch and 5 mg of purified water and a small amount of at least one compound selected from the group consisting of dextrin, maltodextrin, corn starch and microcrystalline cellulose (MCC) as additives for inhibiting hygroscopicity Thereafter, 100 mg of a pellet was prepared according to a conventional method.
<제제예 5. 건강기능식품의 제조>≪ Formulation example 5. Preparation of health functional foods >
본 발명의 실시예 4-1 20g, 비타민 혼합물 적량, 비타민 A 아세테이트 70㎍, 비타민 E 1.0㎎, 비타민 B1 0.13㎎, 비타민 B2 0.15㎎, 비타민 B6 0.5㎎, 비타민 B12 0.2㎍, 비타민 C 10㎎, 비오틴 10㎍, 니코틴산아미드 1.7㎎, 엽산 50㎍, 판토텐산 칼슘 0.5㎎, 무기질 혼합물 적량, 황산제1철 1.75㎎, 산화아연 0.82㎎, 탄산 마그네슘 25.3㎎, 제1인산칼륨 15㎎, 제2인산칼슘 55㎎, 구연산칼륨 90㎎, 탄산칼슘 100㎎, 염화마그네슘 24.8㎎을 섞어 과립으로 제조하였으나, 용도에 따라 다양한 제형으로 변형시켜 제조할 수 있다. 또한, 상기의 비타민 및 미네랄 혼합물의 조성비를 임의로 변형 실시하여도 무방하며, 통상의 건강기능식품 제조방법에 따라 상기의 성분을 혼합하여 제조할 수 있다.20 g of Example 4-1 of the present invention, a proper amount of vitamin A, 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 μg of biotin, 1.7 mg of nicotinic acid amide, 50 μg 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, potassium citrate 90 mg,
<< 제제예Formulation example 6. 건강기능성 음료의 제조> 6. Preparation of Health Functional Drink>
본 발명의 실시예 4-1 1g, 구연산 0.1g, 프락토올리고당 100g, 정제수 900g을 섞어 통상의 음료 제조방법에 따라 교반, 가열, 여과, 살균, 냉장하여 음료를 제조하였다.1 g of Example 4-1 of the present invention, 0.1 g of citric acid, 100 g of fructooligosaccharide, and 900 g of purified water were mixed and stirred, heated, filtered, sterilized and refrigerated according to a conventional beverage preparation method to prepare a drink.
Claims (6)
상기 DNA 단편 혼합물은 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물이 혼합된 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품.The method according to claim 1,
Wherein the DNA fragment mixture is a mixture of a DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa, and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less.
상기 분자량 5kDa이상 50kDa미만인 DNA 단편 혼합물, 분자량 50kDa이상 2500kDa미만인 DNA 단편 혼합물 및 분자량 2500kDa이상 5000kDa이하인 DNA 단편 혼합물은 1~2:1~2:1~2의 중량비로 혼합된 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품. 3. The method of claim 2,
A DNA fragment mixture having a molecular weight of 5 kDa or more and less than 50 kDa, a DNA fragment mixture having a molecular weight of 50 kDa or more and less than 2500 kDa and a DNA fragment mixture having a molecular weight of 2500 kDa or more and 5000 kDa or less are mixed at a weight ratio of 1: 2: 1 to 2: 1 to 2 Health functional foods for prevention or improvement.
상기 DNA 단편 혼합물은 어류의 정액 또는 정소에서 분리한 것임을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품.The method according to claim 1,
Wherein said DNA fragment mixture is isolated from semen or testes of fish.
상기 어류는 연어과의 어류인 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품. 5. The method of claim 4,
Wherein said fish is a fish of salmonid.
상기 관절염은 퇴행성 관절염(osteoarthritis)인 것을 특징으로 하는 관절염의 예방 또는 개선용 건강기능식품.6. The method according to any one of claims 1 to 5,
Wherein said arthritis is degenerative arthritis (osteoarthritis).
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