KR20110122961A - Composition for preventing and treating arthritis comprising hemp seed extract - Google Patents
Composition for preventing and treating arthritis comprising hemp seed extract Download PDFInfo
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- KR20110122961A KR20110122961A KR1020100042339A KR20100042339A KR20110122961A KR 20110122961 A KR20110122961 A KR 20110122961A KR 1020100042339 A KR1020100042339 A KR 1020100042339A KR 20100042339 A KR20100042339 A KR 20100042339A KR 20110122961 A KR20110122961 A KR 20110122961A
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Abstract
Description
본 발명은 헴프 종자 추출물을 포함하는 관절염 예방 및 치료용 조성물에 관한 발명이다.The present invention relates to a composition for the prevention and treatment of arthritis, including the hemp seed extract.
관절염은 크게 골관절염 (퇴행성 관절염, Osteoarthritis)과 류마티스 관절염(Rheumatoid arthritis)으로 구분된다.Arthritis is largely divided into osteoarthritis (Osteoarthritis) and Rheumatoid arthritis.
골관절염은 퇴행성 관절염이라고도 하며 가장 흔히 볼 수 있는 관절염으로 중년 또는 노년에 주로 발생하고, 무릎 관절과 엉덩이 관절과 같이 체중을 많이 받는 관절에 통증, 강직감 및 점진적인 운동장애 등을 초래하는 질환이다.Osteoarthritis, also known as degenerative arthritis, is the most commonly seen arthritis. It occurs mainly in middle age or old age, and causes pain, stiffness, and gradual movement disorder in weight-bearing joints such as knee joints and hip joints.
원인으로는 과거에는 단순히 나이가 들어서 발생하는 노화현상으로 생각하였으나 현재는 여러 가지 원인이 있는 것으로 알려져 있는데, 한가지 원인이 아니라 연령, 유전적 요소, 비만, 관절의 모양, 호르몬 등의 여러 가지 원인이 복합적으로 이루어짐으로써 증상이 다르게 나타나는 것으로 생각되고 있다. 골관절염은 주로 관절연골(cartilage)의 손상에 관여하는데, 연골의 손상에 주로 관여하는 것으로 알려진 매트릭스 메탈로프로테나제 (matrix metalloproteinases, MMPs)가 작용하여 연골조직을 파괴하게 되며, 그 주위의 뼈에 퇴행성 변화를 일으킨다. 또한, 염증성 사이토카인(cytokine)인 인터루킨-1(IL-1), 종양괴사인자-α(TNF-α) 등을 유도시키게 되는데 이로 인하여 근육, 건, 인대 등과 같은 조직에도 영향을 끼쳐 심한 통증을 일으킨다(Fernandes J. C. et al., The role of cytokines in osteoarthritis pathophysiology , 39 , pp237-46, 2002).In the past, as a cause of aging caused by aging, but now it is known that there are many causes, not one cause, but age, genetic factors, obesity, joint shape, hormones, etc. Complications are thought to be different symptoms. Osteoarthritis is primarily involved in cartilage damage. Matrix metalloproteinases (MMPs), known to be involved in cartilage damage, act to destroy cartilage tissue. Causes degenerative changes. In addition, inflammatory cytokines such as interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) are induced, which affects tissues such as muscles, tendons, and ligaments, causing severe pain. (Fernandes JC et al., The role of cytokines in osteoarthritis pathophysiology, 39, pp237-46, 2002).
반면에, 류마티스성 관절염 (Rheumatoid arthritis)은 여러 관절에 다발성으로 나타나는 염증성 자가면역질환이다.Rheumatoid arthritis, on the other hand, is an inflammatory autoimmune disease that occurs multiple times in multiple joints.
관절염에 걸린 환자의 경우, 관절의 활막 (synovial membrane) 조직이 증생(hyperplasia)되며 동시에 대식세포(macrophage), 수상세포(dendritic cell) 및 활성화된 T 림프구와 B 림프구가 활막 조직으로 이동하고 다형핵 세포 (polymorphonuclear cell)는 활액(synovial fluid) 내와 연골(cartilage) 표면에 축적되어 염증을 유발하게 된다. 이같은 활막 조직의 염증은 알려지지 않은 자가항원에 대해 반응성이 있는 T 림프구가 유발하는 것으로 생각되고 있다. 그럼에도 불구하고 대부분의 조직에 침윤된 T 림프구는 세포 표면에 활성화 표지를 나타내지 않으며 사이토카인(cytokine)을 거의 발현하지도 않는다. 이와 대조적으로 류마티스성 관절염 증상을 가진 활막조직과 활액에는 대식세 포 (macrophage)에서 유래된 사이토카인들이 매우 많이 관찰되고 있다. 이러한 사이토카인으로는 활막 섬유 세포의 성장을 촉진하는 것으로 알려진 인터루킨-1 (IL-1), 종양괴사인자-α(TNF-α) 등이 있다. 이러한 실험 결과들은 T 림프구가 활막 조직의 염증을 유발하는 데 매우 중요하며, 이후의 염증 증상은 활성화된 활막 세포들로부터 유래된 사이토카인에 의해 유지된다는 가설을 제시한다(Carson D.A. et al., J. Clin. Invest. , 87 , pp379-383, 1991 : Tighe H. et al., J. Exp. Med. , 177 , pp10-118, 1993 : Burmester G. R. et al., Arthritis Rheum, 40, pp5-18, 1997 : Panayi G. S., Curr. Opin. Rheumatol. , 9 , pp236-240, 1997).In patients with arthritis, synovial membrane tissue of the joints is hyperplasia, while macrophages, dendritic cells and activated T lymphocytes and B lymphocytes migrate to the synovial tissue and polymorphonuclear Polymorphonuclear cells accumulate in synovial fluid and on the surface of cartilage, causing inflammation. Inflammation of these synovial tissues is thought to be caused by T lymphocytes reactive to unknown autoantigens. Nevertheless, T lymphocytes infiltrated most tissues show no activation marker on the cell surface and rarely express cytokines. In contrast, synovial tissue and synovial fluid with rheumatoid arthritis symptoms are very high cytokines derived from macrophage (macrophage). Such cytokines include interleukin-1 (IL-1), tumor necrosis factor-α (TNF-α), which are known to promote the growth of synovial fiber cells. These experimental results suggest that T lymphocytes are very important for causing inflammation of synovial tissue, and that subsequent inflammatory symptoms are maintained by cytokines derived from activated synovial cells (Carson DA et al., J Clin.Invest., 87, pp 379-383, 1991: Tighe H. et al., J. Exp. Med., 177, pp 10-118, 1993: Burmester GR et al., Arthritis Rheum, 40, pp 5-18 , 1997: Panayi GS, Curr. Opin.Rheumatol., 9, pp236-240, 1997).
또한, 활막 조직의 증생은 관절염에 있어서 혈관이 없는 연골부에 발생하는 결합조직층인 판누스 (pannus)를 형성하여 질환 관절의 연골과 골조직을 비가역적으로 파괴하게 되는데, 여기에는 연골조직 파괴인자로 알려진 매트릭스 메탈로프로테나제(MMPs) 등이 작용하여 연골조직이 파괴되게 된다.In addition, synthesizing synovial tissue forms pannus, which is a layer of connective tissue that occurs in cartilage without blood vessels in arthritis, and irreversibly destroys cartilage and bone tissue of diseased joints. Known matrix metalloproteinases (MMPs) and the like act to destroy cartilage tissue.
현재 사용되고 있는 골관절염 및 류마티스 관절염의 치료방법은 다음과 같다. 초기 치료에 많이 사용되는 약물은 비스테로이드성 소염제 (NSAID: non-steroidal anti-inflammatory drug)로서, 이들 NSAID들은 증세를 약간 호전시키기는 하지만 관절부위 연골 손실이나 질병의 진행을 막을 수는 없으며, 부작용이 심하게 나타나 이 치료법을 사용하는 환자의 절반 가량이 1년 이내에 치료를 중단해야만 한다. 다음 단계로는 금을 포함하는 화합물 약제 (gold drugs; gold sodium thiomalate, gold sodium thiosulfate)나 페니실아민 (penicillamine), 항말라리아제 (anti-malarials)와 같은 항관절염 제제 (disease modifying anti-rheumatic drugs; DMARDs) 등이 사용된다. 그러나 이 제재들 역시 관절염의 진행을 다소 감소시키지만 심각한 부작용을 동반할 수 있기 때문에 DMARD를 이용한 치료법이 시작된 5년 이후에는 5-15% 환자만이 이 약물들의 사용을 고수한다. 이상의 치료제가 더 이상 효과를 보이지 않을 경우에는 류마티스가 발생한 관절부위를 외관절 시술에 의해 인공관절로 교체해야만 하는 경우도 있다(Simon L. S., Int. J. Clin. Pract. , 54 , pp243-249, 2000).Currently used treatment methods for osteoarthritis and rheumatoid arthritis are as follows. Drugs commonly used in early treatment are nonsteroidal anti-inflammatory drugs (NSAIDs), which may slightly improve symptoms but do not prevent joint cartilage loss or disease progression. This is severe and about half of patients using this therapy must stop treatment within a year. Next steps include: drugs modifying anti-rheumatic drugs such as gold drugs (gold sodium thiomalate, gold sodium thiosulfate), penicillamine, anti-malarials; DMARDs) and the like. However, these drugs also slightly reduce the progression of arthritis but can have serious side effects, so only 5-15% of patients adhere to the use of these drugs after 5 years of treatment with DMARD. If the above treatments are no longer effective, it may be necessary to replace the joints with rheumatoid joints with external joints (Simon LS, Int. J. Clin. Pract., 54, pp 243-249, 2000).
본 발명은 상기의 문제점을 해결하고 상기의 필요성에 의하여 안출된 것으로서 본 발명의 목적은 관절염 예방 및 치료효과를 가지는 조성물을 제공하는 것이다.The present invention solves the above problems and the object of the present invention as devised by the necessity of the above is to provide a composition having a preventive and therapeutic effect of arthritis.
상기의 목적을 달성하기 위하여 본 발명은 헴프 종자 추출물을 포함하는 관절염 예방 및 치료용 약학 조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for preventing and treating arthritis, including hemp seed extract.
본 발명은 헴프 종자 추출물을 포함하는 관절염 완화용 건강식품 조성물을 제공한다.The present invention provides a health food composition for arthritis relief comprising a hemp seed extract.
본 발명의 또 다른 목적은 도코사헥사엔산(이하 'DHA'라 함)을 유효성분으로 포함하는 관절염 예방 및 치료용 약학 조성물을 제공한다.Another object of the present invention to provide a pharmaceutical composition for the prevention and treatment of arthritis comprising docosahexaenoic acid (hereinafter referred to as 'DHA') as an active ingredient.
본 발명의 또 다른 목적은 도코사헥사엔산을 유효성분으로 포함하는 관절염 완화용 건강식품 조성물을 제공한다.Another object of the present invention to provide a health food composition for arthritis relief comprising a docosahexaenoic acid as an active ingredient.
헴프 종자 추출물의 용매 추출에 사용되는 용매로서는, 본 발명의 관절염의 예방 또는 치료제 작용을 나타내는 물질을 추출할 수 있는 용매라면 어떤 것도 사용할 수 있고, 예를 들면 헥산, 톨루엔, 석유 에테르, 벤젠, 아세트산에틸, 클로로포름, 디클로로메탄, 1,1,2-트리클로로에텐, 디메틸술폭시드, 아세톤 등의 유기 용매 등을 들 수 있고, As a solvent used for solvent extraction of the hemp seed extract, any solvent can be used as long as it can extract a substance exhibiting the prophylactic or therapeutic effect of arthritis of the present invention. For example, hexane, toluene, petroleum ether, benzene, acetic acid Organic solvents such as ethyl, chloroform, dichloromethane, 1,1,2-trichloroethene, dimethyl sulfoxide, acetone, and the like.
완충액으로서는, 예를 들면 인산 완충액, 시트르산 완충액 등을 들 수 있다. 무기 염 수용액의 무기 염으로서는, 예를들면 염화나트륨, 염화갈륨, 염화칼슘 등을 들 수 있다.As a buffer solution, a phosphate buffer, a citric acid buffer, etc. are mentioned, for example. As an inorganic salt of an inorganic salt aqueous solution, sodium chloride, gallium chloride, a calcium chloride, etc. are mentioned, for example.
이들 용매는 단독으로 또는 복수개 혼합하여 사용할 수 있다. These solvent can be used individually or in mixture of multiple pieces.
용매로서는 초임계 유체화한 이산화탄소를 사용할 수도 있다.As the solvent, supercritical fluidized carbon dioxide may also be used.
추출은 예를 들면 식물체 1 중량부에 대하여 용매 0.1 중량부 내지 10000 중량부, 바람직하게는 1 중량부 내지 100중량부를 사용하여 행한다. 추출 온도는 특별히 제한이 없지만, 예를 들면 0 ℃ 내지 100 ℃가 바람직하고, 20 ℃ 내지 90 ℃가 보다 바람직하다. 추출 시간은 특별히 제한이 없지만, 예를 들면 1분간 내지 1 주간이 바람직하고, 30분간 내지 1 일간이 보다 바람직하다.Extraction is performed using 0.1 weight part-10000 weight part, Preferably 1 weight part-100 weight part with respect to 1 weight part of plants, for example. Although extraction temperature does not have a restriction | limiting in particular, For example, 0 degreeC-100 degreeC is preferable, and 20 degreeC-90 degreeC is more preferable. Although extraction time does not have a restriction | limiting in particular, For example, 1 minute-1 week are preferable, and 30 minutes-1 day are more preferable.
유효 성분을 함유하는 추출액은, 유효 성분을 함유하는 식물체로부터 유효 성분이 추출되는 조건에서 각종 용매 추출,초임계 유체 추출 등으로 추출하여 얻어지지만, 상기 추출액을 유효 성분이 추출되는 조건에서 각종 용매 추출, 초임계 유체 추출 등으로 더 추출하여 얻을 수도 있다. 또한, 상기 식물체로부터 유효 성분이 추출되지 않거나, 추출되기 어려운 조건에서 각종 용매 추출, 초임계 유체 추출 등으로 추출하여 얻어지는 추출 잔사 등을, 추가로 유효 성분이 추출되는 조건에서 각종 용매 추출, 초임계 유체 추출 등으로 추출함으로써 제조할 수도 있다.The extract containing the active ingredient is obtained by extracting various solvents, supercritical fluid extraction, etc. under the condition that the active ingredient is extracted from the plant containing the active ingredient, but the extract is extracted under various conditions under which the active ingredient is extracted. It can also be obtained by further extraction, such as supercritical fluid extraction. In addition, the extraction residue obtained by extraction by various solvent extraction, supercritical fluid extraction, etc., under conditions in which the active ingredient is not extracted or difficult to extract from the plant, and various solvent extraction and supercritical conditions under the condition that the active ingredient is extracted. It can also manufacture by extracting by fluid extraction.
추출에 사용되는 기기로서는 특별히 제한이 없지만, 효율적으로 추출하기 위해서 고안된 용기, 교반기, 환류 냉각기,속슬렛 추출기, 균질기, 진탕기, 초음파 발생 장치 등을 들 수 있다.Although there is no restriction | limiting in particular as an apparatus used for extraction, The container, the stirrer, the reflux condenser, the soxhlet extractor, the homogenizer, the shaker, the ultrasonic wave generator, etc. which were devised for efficient extraction are mentioned.
본 발명을 적용할 수 있는 관절염으로서는 특별히 제한이 없고, 클라미디아 관절염, 만성 흡수성 관절염, 장질환성 관절염, 임균성 관절염, 통풍 관절염, 쟈크(Jaccoud) 관절염, 연소기 관절염, 라임 관절염, 알캅톤 뇨성 관절염, 화농성관절염, 변형성 관절증, 견관절 주위염, 과도한 운동 부하에 의한 관절염, 만성 관절 류마티스 등의 어떤 것이어도 상관없지만, 특히 만성 관절 류마티스에 유효하다.There is no restriction | limiting in particular as arthritis to which this invention is applicable, Chlamydia arthritis, chronic absorptive arthritis, enteropathic arthritis, gonococcal arthritis, gouty arthritis, Jaccoud arthritis, combustor arthritis, Lyme arthritis, alcaptonuria arthritis, purulent Arthritis, deformable arthrosis, periarthritis of the shoulder, arthritis due to excessive exercise load, chronic arthritis rheumatoid, etc. may be used, but it is especially effective for chronic arthritis rheumatoid.
본 발명의 관절염의 예방 또는 치료용 약학 조성물, 건강식품, 동물용 사료, 식품 첨가제 및 사료 첨가제에 대하여 이하에 나타낸다.The pharmaceutical composition for preventing or treating arthritis of the present invention, health food, animal feed, food additives and feed additives are shown below.
(1) 본 발명의 추출물 또는 DHA를 유효 성분으로서 함유하는 것을 특징으로 하는 관절염의 예방 또는 치료용 약학 조성물(1) A pharmaceutical composition for the prevention or treatment of arthritis, comprising the extract of the present invention or DHA as an active ingredient
본 발명의 추출물을 유효 성분으로서 함유하는 것을 특징으로 하는 관절염의 예방 또는 치료제는, 상술한 방법으로 제조한 헴프 종자 추출물을 함유하고,필요에 따라서 약리학적으로 허용되는 1종 또는 그 이상의 담체, 추가로 필요에 따라서 다른 예방 또는 치료를 위한 유효 성분을 더 함유할 수도 있다. 또한, 본 발명의 관절염의 예방 또는 치료제에는, 관절염의 예방 또는 치료에 유효한 임의의 다른 성 분이 첨가되어 있을 수도 있다.A prophylactic or therapeutic agent for arthritis, which comprises the extract of the present invention as an active ingredient, contains a hemp seed extract prepared by the above-described method, and optionally, one or more carriers which are pharmacologically acceptable, and further added. As needed, it may further contain an active ingredient for other prophylaxis or treatment. In addition, any other component effective for preventing or treating arthritis may be added to the prophylactic or therapeutic agent for arthritis of the present invention.
관절염의 예방 또는 치료에 유효한 다른 성분(이하, 간단히 다른 유효 성분이라고도 함)으로서는, 예를 들면 붕소, 칼슘, 크롬, 구리, 마그네슘, 망간, 셀렌, 실리콘, 아연, S-아데노실메티오닌, 콜라겐, 콜라겐 가수분해물, 젤라틴, 젤라틴가수분해물, 브로멜라인, 트립신, 키모트립신, 파파인, 루틴, 카로티노이드, 플라보노이드, 항산화 비타민, γ-리놀렌산, 에이코사펜타엔산, 보스웰리아(Boswellia), 캅사이신, 켓츠클로 (Cat's Claw), 악마의 발톱(Devil's Claw), 피버퓨(Feverfew), 생강, 쐐기풀, 나이아시아미드, 상어 연골, 쿠메릭 및 쿠르쿠민 등을 들 수 있고, 이들 성분은 정제물일수도 있고, 상기 성분의 함유물 또는 추출물일 수도 있다.Other ingredients effective for the prevention or treatment of arthritis (hereinafter, simply referred to as other active ingredients) include, for example, boron, calcium, chromium, copper, magnesium, manganese, selenium, silicon, zinc, S-adenosylmethionine, collagen, Collagen hydrolyzate, gelatin, gelatin hydrolysate, bromelain, trypsin, chymotrypsin, papain, rutin, carotenoids, flavonoids, antioxidant vitamins, γ-linolenic acid, eicosapentaenoic acid, boswellia, capsaicin, kets Cat's Claw, Devil's Claw, Feverfew, Ginger, Nettle, Naysiamid, Shark Cartilage, Cumeric and Curcumin, and these components may be purified. It may also be an inclusion or extract of the component.
본 발명의 관절염의 예방제 또는 치료제는 본 발명의 추출물 또는 DHA, 및 필요에 따라서 다른 유효 성분을 약리학적으로 허용되는 1종 또는 그 이상의 담체와 함께 혼합하여, 제제학의 기술 분야에서 잘알려져 있는 임의의 방법에 의해 제조된다.The prophylactic or therapeutic agent for arthritis of the present invention may be mixed with the extract or DHA of the present invention and, if necessary, other active ingredients with one or more pharmacologically acceptable carriers, so as to be known in the art of formulation. It is manufactured by the method.
본 발명의 관절염의 예방제 또는 치료제의 투여 경로는, 관절염의 예방 또는 치료에 효과적인 것을 사용하는 것이 바람직하고, 예를 들면 경구 투여 또는 예를 들면 정맥내 투여 등의 비경구 투여를 들 수 있다.The administration route of the prophylactic or therapeutic agent for arthritis of the present invention is preferably used to prevent or treat arthritis. Examples thereof include parenteral administration such as oral administration or intravenous administration.
투여 형태로서는 예를 들면 정제, 산제, 과립제, 환제, 현탁제, 유제, 침제, 캡슐제, 시럽제, 주사제, 액제, 엘릭시르제,엑기스제, 팅크제, 유동 엑기스제 등을 들 수 있다.Examples of the dosage form include tablets, powders, granules, pills, suspensions, emulsions, emulsifiers, capsules, syrups, injections, solutions, elixirs, extracts, tinctures, liquid extracts, and the like.
경구 투여에 적합한, 예를 들면 시럽제와 같은 액체 제조물은 물, 슈크로오스, 솔비트, 과당 등의 당류, 폴리에틸렌글리콜, 프로필렌글리콜 등의 글리콜류, 참기름, 올리브유, 대두유 등의 기름류 등의 희석제, p-히드록시벤조산에스테르류 등의 방부제, 파라옥시벤조산메틸 등의 파라옥시벤조산 유도체, 벤조산나트륨 등의 보존제, 딸기향, 페퍼민트 등의 향료 등을 사용하여 제조할 수 있다.Suitable liquid preparations for oral administration, for example, syrups include water, sucrose, sorbet, sugars such as fructose, glycols such as polyethylene glycol and propylene glycol, diluents such as oils such as sesame oil, olive oil and soybean oil, preservatives such as p-hydroxybenzoic acid esters, paraoxybenzoic acid derivatives such as methyl paraoxybenzoate, preservatives such as sodium benzoate, and flavors such as strawberry flavor and peppermint.
또한, 경구 투여에 적합한 정제, 산제 및 과립제 등은 유당, 백당, 포도당, 슈크로오스, 만니톨, 소르비톨 등의 당류, 감자, 밀, 옥수수 등의 전분, 탄산칼슘, 황산칼슘, 탄산수소나트륨, 염화나트륨 등의 무기물, 결정 셀룰로오스, 감초 분말,겐티아나 분말 등의 식물 분말 등의 부형제, 전분, 한천, 젤라틴 분말, 결정 셀룰로오스, 카르멜로오스나트륨, 카르멜로오스칼슘, 탄산칼슘, 탄산수소나트륨, 알긴산나트륨 등의 붕해제, 스테아르산마그네슘, 탈크, 수소 첨가 식물유, 마크로골, 실리콘유 등의 활택제, 폴리비닐알코올, 히드록시프로필셀룰로오스, 메틸셀룰로오스, 에틸셀룰로오스, 카르멜로오스, 젤라틴, 전분 풀액 등의 결합제, 지방산 에스테르 등의 계면활성제, 글리세린 등의 가소제 등을 사용하여 제조할 수 있다.In addition, tablets, powders and granules suitable for oral administration include lactose, sugar, glucose, sucrose, saccharides such as mannitol and sorbitol, starch such as potatoes, wheat and corn, calcium carbonate, calcium sulfate, sodium bicarbonate and sodium chloride. Excipients such as inorganic powders, plant powders such as crystalline cellulose, licorice powder, gentian powder, starch, agar, gelatin powder, crystalline cellulose, sodium carmellose, carmellose calcium, calcium carbonate, sodium hydrogencarbonate, sodium alginate Disintegrating agents, such as magnesium stearate, talc, hydrogenated vegetable oil, macrogol, silicone oil and other lubricants, polyvinyl alcohol, hydroxypropyl cellulose, methyl cellulose, ethyl cellulose, carmellose, gelatin, starch liquor, etc. It can manufacture using surfactant, such as a binder and fatty acid ester, and plasticizer, such as glycerin.
비경구 투여에 적합한 제제는, 바람직하게는 수용자의 혈액과 등장인 활성 화합물을 포함하는 멸균 수성제로 이루어진다. 예를 들면, 주사제의 경우에는 염 용액, 포도당 용액 또는 염수와 포도당 용액의 혼합물로 이루어지는 담체 등을 사 용하여 주사용 용액을 제조한다.Formulations suitable for parenteral administration preferably consist of sterile aqueous agents comprising the active compound that is isotonic with the blood of the recipient. For example, in the case of injectables, an injectable solution is prepared using a salt solution, a glucose solution or a carrier composed of a mixture of saline and glucose solution.
또한, 이들 비경구 제제에 있어서도, 경구 제제에서 예시한 희석제, 방부제, 보존제, 향료류, 부형제, 붕해제, 활택제,결합제, 계면활성제, 가소제 등으로부터 선택되는 1종 또는 그 이상의 보조 성분을 첨가할 수도 있다.Also in these parenteral preparations, one or more auxiliary ingredients selected from diluents, preservatives, preservatives, flavors, excipients, disintegrants, lubricants, binders, surfactants, plasticizers and the like exemplified in the oral preparations are added. You may.
본 발명의 관절염 예방제 또는 치료제의 투여량 및 투여 횟수는, 투여 형태, 환자의 연령, 체중, 치료해야 할 증상의 성질 또는 중증도에 의해 다르지만, 통상적인 경구의 경우, 성인 한 사람에 대하여 본 발명의 추출물 또는 DHA의 건조 중량으로서 바람직하게는 0.1 mg 내지 5 g, 보다 바람직하게는 0.5 mg 내지 1 g을 하루 1회 내지 수회 투여한다. 정맥내 투여 등의 비경구 투여의 경우, 성인 한 사람에 대하여 본 발명의 추출물 또는 DHA의 건조 중량으로서 바람직하게는 1mg 내지 5 g, 보다 바람직하게는 5mg 내지 1 g을 하루 1회 내지 수회 투여한다.The dosage and frequency of administration of the arthritis prophylactic or therapeutic agent of the present invention vary depending on the dosage form, the age, weight of the patient, the nature or severity of the symptom to be treated, and in the case of conventional oral treatment, As dry weight of the extract or DHA, preferably 0.1 mg to 5 g, more preferably 0.5 mg to 1 g is administered once to several times a day. In the case of parenteral administration such as intravenous administration, preferably, 1 mg to 5 g, more preferably 5 mg to 1 g is administered once or several times as dry weight of the extract or DHA of the present invention to one adult. .
본 발명의 관절염의 예방제를 일상적으로 투여함으로써 관절염을 예방할 수 있다. 이미 관절염이 발병되어 있는 경우에는, 본 발명의 관절염의 치료제를 일상적으로 투여함으로써 관절염의 증상을 완화, 치유하는 등, 관절염을 치료할 수 있다. 본 발명의 관절염의 예방제 또는 치료제의 투여 기간은 통상 1주간 내지 10년간, 바람직하게는 1개월간 내지 5년간이다.Arthritis can be prevented by routinely administering the prophylactic agent for arthritis of the present invention. If arthritis has already developed, arthritis can be treated by routinely administering the therapeutic agent for arthritis of the present invention to alleviate and cure symptoms of arthritis. The administration period of the prophylactic or therapeutic agent for arthritis of the present invention is usually 1 week to 10 years, preferably 1 month to 5 years.
또한, 본 발명의 관절염의 예방제 또는 치료제는 사람 뿐만 아니라 사람이외의 동물에 대해서도 사용할 수 있다. 동물에게 사용하는 경우의 투여량은 특별히 제한은 없지만, 본 발명의 추출물의 건조 중량으로서 바람직하게는 0.01 내지 5 g/kg이다.In addition, the prophylactic or therapeutic agent for arthritis of the present invention can be used not only for humans but also for animals other than humans. Although the dosage in the case of using for an animal is not restrict | limited, Preferably it is 0.01-5 g / kg as dry weight of the extract of this invention.
(2) 본 발명의 헴프 종자 추출물 또는 DHA를 유효 성분으로서 함유하는 것을 특징으로 하는 관절염의 예방 또는 치료용 건강식품 또는 사료(2) Health food or feed for the prevention or treatment of arthritis, comprising hemp seed extract or DHA of the present invention as an active ingredient
본 발명의 추출물을 유효 성분으로서 함유하는 것을 특징으로 하는 관절염의 예방 또는 치료용 식품 또는 사료로서는 본 발명의 추출물 또는 DHA를 그대로 식품 또는 사료로 만든 것, 또는 상기 추출물 또는 DHA를 식품 또는 사료에 첨가한 것을 들 수 있다. 상기 식품 또는 사료는 건강 식품, 기능성 식품으로서 관절염의 예방 또는 치료를 위해 사용된다.Food or feed for prevention or treatment of arthritis, comprising the extract of the present invention as an active ingredient, the extract or DHA of the present invention as a food or feed as it is, or the extract or DHA is added to food or feed One thing is possible. The food or feed is used for the prevention or treatment of arthritis as a health food, a functional food.
본 발명의 식품 또는 사료는, 본 발명의 추출물, 또는 필요에 따라서 다른 유효 성분을 첨가하는 것 이외에는 일반적인 식품 또는 사료의 제조 방법을 이용함으로써 제조할 수 있다.The food or feed of the present invention can be produced by using a general food or feed production method except adding the extract of the present invention or other active ingredients as necessary.
또한, 상기 식품 또는 사료는 일반적인 식품 또는 사료를 예를 들면 성형ㆍ조립 등의 가공 방법으로 가공할 수도 있다. 가공 방법으로서는 유동층 조립, 교반 조립, 압출 조립, 전동 조립, 기류 조립, 압축 성형 조립, 파쇄 조립, 분무조립, 분사 조립 등의 조립 방법, 팬 코팅, 유동층 코팅, 건식 코팅 등의 코팅 방법, 퍼프 드라이, 과잉 수증기법, 폼 매트 방법, 마이크로파 가열 방법 등의 팽화 방법 등을 들 수 있다.The food or feed may also be processed into a general food or feed by a processing method such as molding and assembling. Processing methods include fluidized bed granulation, stirring granulation, extrusion granulation, electric granulation, airflow granulation, compression molding granulation, crushing granulation, spray granulation, granulation methods such as spray granulation, coating methods such as fan coating, fluidized bed coating and dry coating, and puff dry. And expansion methods such as an excess water vapor method, a foam mat method, and a microwave heating method.
본 발명의 식품 또는 사료에 첨가하는 본 발명의 추출물은 본 발명의 식품 또는 사료가 관절염의 예방 또는 치료 작용을 갖는 것이라면, 첨가량에 특별히 제한은 없지만, 예를 들면 0.01 내지 50 중량%, 바람직하게는 0.1 내지 10 중량%를 함유하도록 첨가된다.The extract of the present invention added to the food or feed of the present invention is not particularly limited as long as the food or feed of the present invention has a prophylactic or therapeutic effect of arthritis, for example, 0.01 to 50% by weight, preferably It is added to contain 0.1 to 10% by weight.
식품에 본 발명의 추출물 또는 DHA를 첨가하여 얻어지는 식품으로서는 예를 들면 본 발명의 추출물을 첨가한 건강 식품, 건강 음료, 쥬스류, 청량 음료수,스프류, 차류, 유산균 음료, 발효유, 빙과, 버터, 치즈, 요구르트, 가공유, 탈지 분유 등의 유제품, 햄, 소세지, 햄버거 등의 축육 제품, 어육 혼합 제품, 계란말이, 달걀 두부 등의 알제품, 쿠키, 젤리, 스낵 과자, 츄잉껌 등의 과자류, 빵류, 국수류, 김치류, 훈제품, 건물, 해산물 조림, 조미료 등을 들 수 있다.Examples of the foods obtained by adding the extract of the present invention or DHA to foods include, for example, health foods, health drinks, juices, soft drinks, soups, teas, lactobacillus drinks, fermented milk, ice cream, butter, and cheese. Dairy products such as yogurt, processed milk, skim milk powder, meat products such as ham, sausages, hamburgers, fish meat products, egg rolls, egg tofu products, cookies, jelly, snack cookies, chewing gum and other confectionery, breads, noodles, Kimchi, smoked products, buildings, boiled seafood, seasonings.
식품의 형태로서는 본 발명의 추출물 또는 DHA를 함유하고 있다면 특별히 제한은 없고, 예를 들면 분말 식품, 시트상 식 품, 병조림 식품, 통조림 식품, 레토르트 식품, 캡슐 식품, 정제상 식품, 유동 식품, 드링크제 등을 들 수 있다.There is no restriction | limiting in particular if it contains the extract of this invention or DHA as a form of food, For example, powdered food, sheet food, canned food, canned food, retort food, capsule food, tablet food, liquid food, drink Etc. can be mentioned.
본 발명의 식품에는 필요에 따라서 일반적인 식품에 사용되는 감미료, 착색료, 보존료, 증점 안정제, 산화 방지제,발색제, 표백제, 곰팡이 방지제, 검베이스, 고미료, 효소, 광택제, 산미료, 조미료, 유화제, 강화제, 제조용 제제, 향료,향신료 추출물 등의 첨가물을 첨가할 수도 있다.In the food of the present invention, sweeteners, coloring agents, preservatives, thickeners, antioxidants, coloring agents, bleaches, fungicides, gum bases, high seasonings, enzymes, varnishes, acidulants, seasonings, emulsifiers, reinforcing agents, Additives, such as a preparation for preparation, a fragrance, and a fragrance extract, can also be added.
감미료로서는 아스파탐, 감초, 스테비아, 크실로오스 등을 들 수 있다.As sweetener, aspartame, licorice, stevia, xylose, etc. are mentioned.
착색료로서는 카로티노이드나 쿠르쿠민 색소, 후라보노이드, 카라멜 색소, 시콘 색소, 스피룰리나 색소, 엽록소, 퍼플스윗 감자(Purple sweet potato) 색소, 퍼플 스윗 산감자 색소, 들깨(Perilla) 색소, 블루베리 색소 등을 들 수 있다.Examples of the colorant include carotenoids, curcumin pigments, flavonoids, caramel pigments, sycon pigments, spirulina pigments, chlorophyll, purple sweet potato pigments, purple sweet acid potato pigments, perilla pigments, and blueberry pigments. .
보존료로서는 아황산나트륨, 벤조산류, 독활(aralia cordata) 추출물, 에고노기 추출물, 사철쑥 추출물, 소르빈산류,프로피온산류 등을 들 수 있다.Examples of the preservative include sodium sulfite, benzoic acid, aralia cordata extract, egonogi extract, cedar extract, sorbic acid and propionic acid.
증점 안정제로서는 아라비아 검이나 크산탄 검 등의 검류, 알긴산류, 키틴, 키토산, 알로에 추출물, 구아 검, 히드록시프로필셀룰로오스, 카제인나트륨, 옥수수 전분, 카르복시메틸셀룰로오스류, 젤라틴, 한천, 덱스트린, 메틸셀룰로오스,폴리비닐알코올, 미소 섬유상 셀룰로오스, 미결정 셀룰로오스, 해초 셀룰로오스, 폴리아크릴산 소다, 폴리인산나트륨,카라기난, 효모 세포벽, 곤약 감자 추출물, 나타드코코(Nata de coco), 만난 등을 들 수 있다.Examples of thickeners include gums such as gum arabic and xanthan gum, alginic acid, chitin, chitosan, aloe extract, guar gum, hydroxypropyl cellulose, sodium casein, corn starch, carboxymethylcellulose, gelatin, agar, dextrin and methylcellulose. , Polyvinyl alcohol, microfibrous cellulose, microcrystalline cellulose, seaweed cellulose, soda polyacrylate, sodium polyphosphate, carrageenan, yeast cell wall, konjac potato extract, Nata de coco, mannan and the like.
산화 방지제로서는 비타민 C, 에틸렌디아민 사아세트산나트륨, 에틸렌디아민 사아세트산칼슘, 에리소르빈산, 오리자놀, 카테친(catechin), 퀘르세틴(quercetin), 클로브 추출물, 효소 처리 루틴, 사과 추출물, 참기름 추출물, 디부틸히드록시톨루엔, 회향 추출물, 서양고추냉이 추출물, 미나리 추출물, 차 추출물, 템페 추출물, 약모밀 추출물, 토코트리에놀, 토코페롤류, 평지 기름 추출물, 원두 커피 추출물, 해바라기 종자, 페룰산, 부틸히드록시아니솔, 블루베리잎 추출물, 프로폴리스 추출물, 고사리ㆍ은행나무 추출물, 헤스페리틴, 후추 추출물, 봉선화 추출물, 갈산, 소귀나무 추출물,유칼립투스 추출물, 로즈마리 추출물 등을 들 수 있다.As antioxidants, vitamin C, ethylenediamine sodium tetraacetate, ethylenediamine calcium tetraacetate, erythorbic acid, oryzanol, catechin, quercetin, clove extract, enzyme treatment routine, apple extract, sesame oil extract, dibutylhydroxy Toluene, Fennel Extract, Horseradish Extract, Buttercup Extract, Tea Extract, Tempe Extract, Hair Buckwheat Extract, Tocotrienol, Tocopherols, Colza Oil Extract, Ground Coffee Extract, Sunflower Seed, Ferulic Acid, Butylhydroxyanisole, Blueberry Leaf Extracts, propolis extracts, fern and ginkgo extracts, hesperidin, pepper extracts, balsam extracts, gallic acid, locust extracts, eucalyptus extracts, rosemary extracts and the like.
발색제로서는 아질산나트륨 등을 들 수 있다.표백제로서는 아황산나트륨 등을 들 수 있다.곰팡이 방지제로서는 오르토페닐페놀 등을 들 수 있다.Sodium nitrite etc. are mentioned as a coloring agent. Sodium sulfite etc. are mentioned as a bleaching agent. Ortho phenyl phenol etc. are mentioned as a mold inhibitor.
껌베이스로서는 아세틸리시놀산메틸, 옻 왁스, 에스테르 검, 엘레미(elemi) 수지, 오우리큐우릴로우, 오조케라이트(Ozokerite), 오포파낙스(Opopanax) 수지, 카우리 검, 카르나우바 왁스, 구아이악(guaiac) 수지, 구타카츄, 구타한칸,구타페르카(Guttapercha), 글리세린지방산 에스테르, 고래 왁스, 코파오바 발삼, 코팔(copal) 수지, 고무, 라이스 왁스, 사탕수수 왁스, 쉘락, 젤톤, 슈크로오스지방산 에스테르, 소르바, 소르비탄산지방산 에스테르, 탈크, 탄산칼슘, 담마(Dammar) 검, 치클(chicle), 치르테, 쯔누(tunu), 저분자 고무, 파라핀 오일, 퍼 발삼, 프로필렌글리콜지방산 에스테르, 분말 펄프, 분말 벼겨, 호호바 오일, 폴리이소부틸렌, 폴리부텐, 마이크로크리스탈 왁스, 유향수(mastic), 맛사란드쵸콜 레이트, 황랍, 인산칼슘 등을 들 수 있다.Examples of the gum base are methyl acetyl ricinoleate, lacquer wax, ester gum, elemi resin, urikyuurilow, ozokerite, oppopanax resin, kauri gum, carnauba wax, and guar. Guiac resins, guttachuchu, guttahankan, guttapercha, glycerin fatty acid esters, whale wax, copaova balsam, copal resin, rubber, rice wax, sugar cane wax, shellac, geltone, Sucrose fatty acid ester, sorbate, sorbitan fatty acid ester, talc, calcium carbonate, dammar gum, chicle, thyrte, tunu, low molecular rubber, paraffin oil, per balsam, propylene glycol Fatty acid esters, powdered pulp, powdered rice bran, jojoba oil, polyisobutylene, polybutene, microcrystal wax, frankincense, masticane chocolate, yellow wax, calcium phosphate and the like.
고미료로서는 이소 알파 고미산, 카페인, 구름버섯 추출물, 키나(kina) 추출물, 황다랑어 추출물, 겐티아나 추출물, 향신료 추출물, 효소 처리 나린진, 쟈마이카 카시아 추출물, 데오브로민, 나린진, 비가끼 추출물, 약쑥 추출물, 방아풀 추출물, 아가리쿠스 버섯, 버섯 추출물, 보라페트, 메틸티오아데노신, 영지 추출물, 올리브차, 광귤 추출물, 홉 추출물,머그워트(mugwort) 추출물 등을 들 수 있다.As high-fertilizers, iso alpha gomic acid, caffeine, cloud mushroom extract, kina extract, yellowfin tuna extract, gentiana extract, spice extract, enzyme treatment naringin, jamaica cassia extract, deobromine, naringin, rain moss extract, Wormwood extract, plant extract, agaricus mushroom, mushroom extract, boraphet, methylthio adenosine, ganoderma lucidum extract, olive tea, tangerine extract, hop extract, mugwort extract and the like.
효소로서는 아밀라아제, 트립신, 린넷, 유산균 등을 들 수 있다.Examples of enzymes include amylase, trypsin, linnet, and lactic acid bacteria.
광택제로서는 옻 왁스, 목랍 등을 들 수 있다.Lacquer wax, wax, etc. are mentioned as a brightening agent.
산미료로서는 아디프산, 이타콘산, 시트르산류, 숙신산류, 아세트산나트륨, 타르타르산류, 이산화탄소, 락트산, 피틴산, 푸마르산, 말산, 인산 등을 들 수 있다.Examples of the acidulant include adipic acid, itaconic acid, citric acid, succinic acid, sodium acetate, tartaric acid, carbon dioxide, lactic acid, phytic acid, fumaric acid, malic acid and phosphoric acid.
조미료로서는 아스파라긴, 아스파라긴산류, 글루탐산, 글루타민, 알라닌, 이소류신, 글리신, 세린, 시스틴, 티로신, 류신, 프롤린 등의 아미노산, 이노신산나트륨, 우리딜산나트륨, 구아닐산나트륨, 시티딜산나트륨, 리보뉴클레오티드 칼슘, 리보뉴클레오티드 나트륨 등의 핵산, 시트르산, 숙신산 등의 유기 산, 염화칼륨, 염수 호수 저염 나트륨액, 조제(粗製) 해수 염화칼륨, 유청 미네랄, 인산삼칼륨, 인산수소이칼륨, 인산이수소칼륨, 인산수소이나트륨,, 인산이수소나트륨, 인산삼나트륨, 클로렐라 추출물 등을 들 수 있다.As seasoning, asparagine, aspartic acid, glutamic acid, glutamine, alanine, isoleucine, glycine, serine, cystine, tyrosine, leucine, amino acids such as proline, sodium inosine, sodium urinate, sodium guanylate, sodium citrate, ribonucleotide calcium, ribonucleotide Nucleic acids such as sodium, organic acids such as citric acid and succinic acid, potassium chloride, salt lake low salt sodium salt, prepared sea salt potassium chloride, whey mineral, tripotassium phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, disodium hydrogen phosphate, phosphoric acid Sodium dihydrogen, trisodium phosphate, chlorella extract and the like.
유화제로서는 지방산 모노글리세라이드, 소르비탄 지방산 에스테르 등을 들 수 있다.Examples of the emulsifier include fatty acid monoglycerides and sorbitan fatty acid esters.
강화제로서는 아연 염류, 비타민 C, 각종 아미노산, 5-아데닐산, 염화철, 헤스페리딘, 각종 소성 칼슘, 각종 미소성 칼슘, 디벤조일티아민, 수산화칼슘, 탄산칼슘, 티아민염산염, 듀나리에라 카로틴, 토코페롤, 니코틴산, 당근 카로틴, 팜유카로틴, 판토텐산칼슘, 비타민 A, 히드록시프롤린, 피롤린산이수소칼슘, 피롤린산 제1철, 피롤린산 제2철, 페리틴, 햄철, 메나퀴논, 엽산, 리보플라빈 등을 들 수 있다.Enhancers include zinc salts, vitamin C, various amino acids, 5-adenylic acid, iron chloride, hesperidin, various calcined calcium, various uncalculated calcium, dibenzoylthiamine, calcium hydroxide, calcium carbonate, thiamine hydrochloride, dunariera carotene, tocopherol, nicotinic acid, carrot Carotene, palm eucarotene, calcium pantothenate, vitamin A, hydroxyproline, calcium dihydrogen pyrroline, ferrous pyrroline, ferric pyrolate, ferritin, ham iron, menaquinone, folic acid, riboflavin and the like.
제조용 제제로서는 아세톤, 이온 교환 수지 등의 가공 조제, 무화과 나뭇잎 추출물, 볏짚 재 추출물, 카올린, 글리세린지방산 에스테르, 뽕나무 추출물, 골회, 들깨 추출물, 생강 추출물, 각종 탄닌, 파파야 추출물, 포도 종자 추출물, 에탄Examples of preparations include acetone, processing aids such as ion exchange resins, fig leaf extract, rice straw extract, kaolin, glycerin fatty acid ester, mulberry extract, bone ash, perilla extract, ginger extract, various tannins, papaya extract, grape seed extract, ethane
올 등을 들 수 있다.향료로서는 상술한 향료 등을 들 수 있다.Or the like. Examples of the fragrance include the fragrance described above.
향신료 추출물로서는 고추 추출물, 마늘 추출물 등을 들 수 있다.Pepper extract, garlic extract, etc. are mentioned as a spice extract.
상기 식품은 1 일에 1회 또는 수회로 나누어 섭취한다. 상기 음식품을 일상적으로 섭취함으로써 관절염을 예방할 수 있다. 이미 관절염이 발병되어 있는 경우에는, 상기 음식품을 일상적으로 섭취함으로써 관절염의 증상을 완화, 치유시키는 등, 관절염을 치료할 수 있다.The food is taken once or divided into several times a day. Arthritis can be prevented by eating the food and drink routinely. If arthritis has already developed, arthritis can be treated by daily ingestion of the food and drink, thereby alleviating and healing the symptoms of arthritis.
본 발명의 추출물을 첨가하여 이루어지는 사료는, 포유류, 조류,파충류, 양서류, 어류 등의 동물, 바람직하게는 포유류에 대한 사료로서 사용된다. 예를 들면, 본 발명의 사료는 개, 고양이, 쥐 등의 애완 동물용 사료, 소, 돼지 등의 가축용 사료, 닭, 칠면조 등의 가금용 사료, 도미, 방어 등의 양식어용 사료 등으로서 사용되고, 바람직하게는 애완 동물용 사료 또는 가축용 사료로서 사용된다.The feed obtained by adding the extract of the present invention is used as a feed for animals such as mammals, birds, reptiles, amphibians, fish, preferably mammals. For example, the feed of the present invention is used as feed for pets such as dogs, cats, rats, cattle feed such as cows and pigs, feed for poultry such as chickens and turkeys, feed for breeding fish such as sea breams and defenses , Preferably used as feed for pets or feed for livestock.
본 발명의 사료는 사료 원료에 본 발명의 추출물을 적절하게 배합하여 만들 수 있다. 사료 원료로서는 곡물류, 조강류, 식물성 유박류, 동물성 사료 원료, 기타 사료 원료, 정제품 등을 들 수 있다.The feed of the present invention can be made by appropriately blending the extract of the present invention with feed ingredients. Examples of feed materials include cereal grains, crude steel, vegetable oils, animal feedstuffs, other feedstuffs, and refined products.
곡물류로서는, 예를 들면 마이로, 밀, 보리, 귀리, 호밀, 현미, 메밀, 조, 수수, 피, 옥수수, 대두 등을 들 수 있다.Examples of cereals include maize, wheat, barley, oats, rye, brown rice, buckwheat, buckwheat, sorghum, blood, corn, soybeans and the like.
조강류로서는, 예를 들면 쌀겨, 탈지 쌀겨, 밀기울, 말분, 밀, 배아, 보리겨, 스크리닝, 펠렛, 옥수수겨, 옥수수 배아 등을 들 수 있다.Examples of the crude steel include rice bran, defatted rice bran, wheat bran, horse meal, wheat, embryo, barley bran, screening, pellets, corn bran, and corn embryo.
식물성 유박류로서는, 예를 들면 대두박, 콩가루, 아마박, 면실박, 낙화생 박, 홍화박, 야자박, 팜박, 호마박, 해바라기박, 유채박, 케이폭박, 겨자박 등을 들 수 있다.Examples of vegetable oils include soybean meal, soybean meal, flaxseed meal, cottonseed meal, groundnut meal meal, safflower meal, palm oil palm, palm oil, fomarak, sunflower oil, rapeseed meal, cowpea oil and mustard oil.
동물성 사료 원료로서는, 예들 들면 어분(북양 밀, 수입 밀, 홀 밀, 연안 밀), 피쉬솔불, 육분, 어골분, 혈분, 분해모, 골분, 가축용 처리 부산물, 페더 밀, 누에, 탈지 분유, 카제인, 건조 유장 등을 들 수 있다.As animal feed materials, for example, fish meal (North American wheat, imported wheat, whole wheat, coastal wheat), fish bulrush, meat meal, fish bone meal, blood meal, decomposing hair, bone meal, animal by-products, feather mill, silkworm, skim milk powder, Casein, dried whey, and the like.
그 밖의 사료 원료로서는 식물 경엽류(알팔파, 헤이큐브, 알팔파잎 분말, 유사 아카시아 분말 등), 옥수수 가공 공업부산물(콘 글루텐, 밀, 콘 글루텐 피드, 콘스테이프리카 등), 전분 가공품, 설탕, 발효 공업 산물(효모, 맥주박, 맥아근,알코올박, 장유박 등), 농산 제조 부산물(감귤 가공박, 두부박, 커피박, 코코아박 등), 그 외(카사바, 잠두콩, 구아밀, 해조, 크릴, 스피룰리나, 클로렐라, 광물 등) 등을 들 수 있다.Other feedstocks include plant foliage (alfalfa, haycube, alfalfa leaf powder, pseudo acacia powder, etc.), corn processing industrial by-products (corn gluten, wheat, corn gluten feed, corn staple, etc.), starch processed products, sugar, fermentation. Industrial products (yeast, beer foil, malt sprouts, alcohol foil, walnuts, etc.), agricultural production by-products (citrus fruits, tofu, coffee, cocoa, etc.), others (cassava, bean, guam, seaweed, Krill, spirulina, chlorella, minerals, etc.) etc. are mentioned.
정제품으로서는 단백질(카제인, 알부민 등), 아미노산, 당질(전분, 셀룰로오스, 슈크로우스, 글루코오스 등), 미네랄,비타민 등을 들 수 있다.Typical products include proteins (casein, albumin, etc.), amino acids, sugars (starch, cellulose, sucrose, glucose, etc.), minerals, vitamins, and the like.
본 발명의 동물용 사료를 동물에게 일상적으로 섭취시킴으로써 관절염을 예방할 수 있다.Arthritis can be prevented by daily ingestion of the animal feed of the present invention in animals.
이미 관절염이 발병되어 있는 경우에는, 상기 동물용 사료를 일상적으로 섭취시킴으로써 관절염의 증상을 완화, 치유시키는 등, 관절염을 치료할 수 있다.If arthritis has already developed, arthritis can be treated by daily ingestion of the animal feed to alleviate and cure symptoms of arthritis.
이하 본 발명을 설명한다.Hereinafter, the present invention will be described.
본 발명의 주요 내용은 Hemp 종자 추출물 및 DHA에 의한 류마티스 관절염 활막세포의 조절이다. 류마티스 관절염 활막세포는 비정상적인 증식을 통해 염증반응을 일으키고 결국 연골과 뼈의 파괴를 유도한다. 따라서 활막세포의 증식 억제나 세포사멸 유도를 통해 류마티스 관절염의 발병 억제 및 치료 효과를 기대할 수 있다. 본 발명에서는 Hemp 종자 추출물 및 그 주요 성분인 DHA가 류마티스 환자의 활막세포주인 MH7A 세포의 증식과 분화, 그리고 세포사멸을 조절하는 기능이 있음을 증명하였다. Hemp 종자 추출물은 MH7A 세포의 증식을 억제함과 동시에 분화를 촉진하는 작용을 하고, Hemp 종자 추출물의 유효 성분인 DHA는 MH7A 세포의 이상 증식 억제 (CCK-8 assay 결과), 세포 사멸 유도 (Western 분석 결과), 활막세포의 분화 유도 (Oil red O 염색법에 의한 분석) 작용이 있다.The main subject of the present invention is the regulation of rheumatoid arthritis synovial cells by Hemp seed extract and DHA. Rheumatoid arthritis synovial cells cause inflammatory reactions through abnormal proliferation, leading to destruction of cartilage and bone. Therefore, anti-proliferation of synovial cells or induction of apoptosis can be expected to inhibit the development and treatment of rheumatoid arthritis. In the present invention, Hemp seed extract and its main ingredient, DHA, proved to have a function of regulating proliferation, differentiation, and apoptosis of MH7A cells, which are synovial cell lines of rheumatoid patients. Hemp seed extract inhibits the proliferation of MH7A cells and promotes differentiation, while DHA, an active ingredient of Hemp seed extract, inhibits abnormal proliferation of MH7A cells (from CCK-8 assay) and induces cell death (Western analysis). As a result, it induces differentiation of synovial cells (analysis by oil red O staining).
본 발명을 통해서 알 수 있는 바와 같이 본 발명의 조성물은 관절염 환자의 발병 방지 및 증상 완화를 위한 치료에 응용될 수 있다. As can be seen from the present invention, the composition of the present invention can be applied to the treatment for preventing the onset and symptom relief of arthritis patients.
도 1은 Hemp seed oil 추출물에 의한 MH7A 세포주의 증식 억제 효과를 확인한 그림으로, (1a)는 Cell counting Kit-8 assay에 의한 세포 증식 억제 효과 확인, (1b)는 DAPI 염색법에 의한 염색체 응축여부 확인, 그리고 (1c)는 Oil red O 염색법에 의한 세포 분화정도를 확인한 그림이다.
도 2는 DHA에 의한 MH7A 세포주의 이상증식 억제 효과를 확인한 그림으로, (2a)는 Cell counting Kit-8 assay에 의한 세포증식 억제 확인, (2b)는 Western blot에 의한 PARP cleavage 확인, (2c)는 DAPI 염색법에 의한 염색체 응축여부를 확인한 그림이다.1 is a diagram confirming the inhibitory effect of the growth of MH7A cell line by Hemp seed oil extract, (1a) confirmed the cell proliferation inhibitory effect by Cell counting Kit-8 assay, (1b) confirmed the chromosome condensation by DAPI staining method , And (1c) shows the differentiation of cells by Oil red O staining.
2 is a diagram confirming the effect of inhibiting aberrant growth of MH7A cell line by DHA, (2a) confirmed the cell proliferation inhibition by Cell counting Kit-8 assay, (2b) confirmed the PARP cleavage by Western blot, (2c) Shows the chromosome condensation by DAPI staining.
이하 비한정적인 실시예를 통하여 본 발명을 더욱 상세하게 설명한다. 단 하기 실시예는 본 발명을 예시하기 위한 목적으로 기재한 것으로서 본 발명의 범위는 하기 실시예에 의하여 제한되는 것으로 해석되지 아니한다.The present invention will now be described in more detail by way of non-limiting examples. However, the following examples are described for the purpose of illustrating the present invention and the scope of the present invention is not to be construed as being limited by the following examples.
실시예Example 1: One: 헴프Hemp 종자 추출물의 관절염에 대한 효과 Effect of Seed Extract on Arthritis
Hemp 종자(충남 당진군 청삼영농조합)를 압착하여 얻은 Hemp 종자유를 Hexane을 이용해 Hemp 종자 Hexane 추출물을 분리하였다. 분리한 Hemp 종자 Hexane 추출물 1ml에 100% pure grade Ethanol 2 ml을 넣고 상온에서 Vortex를 이용해 1분간 섞는다. 섞인 Hexane+Ethanol 용액을 상온에서 12000rpm으로 2분간 원심분리한다. 원심분리 후 상층 (Ethanol층)을 분리해 실험에 사용하였다. Hemp seed oil obtained by pressing Hemp seeds (Cheongsam farming association of Chungjin Dangjin-gun, Chungnam) was separated using Hexane. 2 ml of 100% pure grade ethanol was added to 1 ml of the Hemp seed Hexane extract isolated, and mixed at room temperature for 1 minute using Vortex. The mixed Hexane + Ethanol solution is centrifuged at 12000 rpm for 2 minutes at room temperature. After centrifugation, the upper layer (Ethanol layer) was separated and used for the experiment.
MH7A, 류마티스 관절염 환자에서 유래된 활막세포 (human fibroblast-like synoviocytes) 세포주는 10 % heat-inactivated fetal bovine serum과 1 % penicillin/streptomycin mitures (Invitrogen, USA)가 포함된 RPMI 1640배지에서 5 % CO2, 37 ℃ 조건으로 배양되었다.MH7A, a human fibroblast-like synoviocytes cell line derived from patients with rheumatoid arthritis, 5% CO2 in RPMI 1640 medium containing 10% heat-inactivated fetal bovine serum and 1% penicillin / streptomycin mitures (Invitrogen, USA) The cells were incubated at 37 ° C.
본 발명의 Hemp 종자 추출물이 MH7A 세포주에 미치는 영향을 알아보기 위해 MH7A에 Hemp 종자 추출물을 농도별로 처리하였다(도 1). MH7A 세포를 48 well plate에 seeding하고 16시간 후 세포량이 60 % 정도일 때 PBS로 2번 워싱하고 Hemp 종자 추출물을 농도별로 섞은 RPMI 1640 배지 2 ml를 각 well에 처리하였다. 5 % CO2, 37 ℃ 조건에서 24시간 배양한 후 Cell counting kit-8의 방법으로 각 well에 10% cck-8 solution을 넣고, 37℃, CO2 5%의 환경의 incubator에서 1시간 동안 incubation 시켰다. 이후에 microplate reader를 이용해서 450/690 nm 파장대를 측정하고 수치를 그래프로 나타내어 각각에 대한 세포의 생존 정도를 파악하였다. 실험결과 Hemp 종자 추출물에 의해 MH7A 세포수가 감소된 것을 확인하였다 (도 1a). In order to determine the effect of the Hemp seed extract of the present invention on the MH7A cell line, Hemp seed extract was treated by MH7A by concentration (FIG. 1). MH7A cells were seeded in 48 well plates, and after 16 hours, when the cell volume was about 60%, the cells were washed twice with PBS and treated with 2 ml of RPMI 1640 medium containing Hemp seed extracts. After incubation for 24 hours at 5% CO 2 , 37 ℃, put 10% cck-8 solution in each well by the method of Cell counting kit-8, incubation for 1 hour in an incubator of 37 ℃, 5% CO 2 environment I was. Afterwards, the 450/690 nm wavelength band was measured using a microplate reader, and the values were graphed to determine the survival rate of the cells for each. Experimental results showed that the MH7A cell number was reduced by the Hemp seed extract (FIG. 1A).
Hemp 종자 추출물이 MH7A 세포에서 Apoptosis를 유도했는지 알아보기 위해 DAPI 염색법을 통해 염색체 응축여부를 확인하였다 (도 1b). MH7A 세포를 6 well plate에 seeding하고 16시간 후 세포량이 60 % 정도일 때 PBS로 2번 워싱하고 Hemp 종자 추출물을 농도별로 섞은 RPMI 1640 배지 2 ml를 각 well에 처리하였다. 5 % CO2, 37 ℃ 조건에서 24시간 배양한 후 inverted microscopy에서 400배 확대한 세포사진을 찍고 PBS 2 ml로 2번 워싱하였다. 4% paraformaldehyde 500 μl를 넣어 상온에서 20분간 고정하고 PBS 2 ml로 2번 워싱했다. DAPI 1 mg/ml stock을 PBS에 1/1000으로 희석하여 세포에 1ml씩 처리하고 5분 후 PBS 1 ml을 더해주고 형광 현미경을 이용해 400배에서 염색된 핵을 관찰하고 사진을 찍었다. 실험 결과 염색체 응축 현상이 일어나지 않아 세포수 감소가 Apoptosis 결과 나타난 현상이 아니라는 것을 알 수 있었다 (도 1b).To determine whether Hemp seed extract induced Apoptosis in MH7A cells, chromosome condensation was confirmed by DAPI staining (FIG. 1B). MH7A cells were seeded in 6 well plates, and after 16 hours, when the cell volume was about 60%, the cells were washed twice with PBS and treated with 2 ml of RPMI 1640 medium containing Hemp seed extract at different concentrations. After culturing for 24 hours at 5% CO 2 , 37 ° C, 400 times magnified cell photographs were taken by inverted microscopy and washed twice with 2 ml of PBS. 500 μl of 4% paraformaldehyde was added and fixed at room temperature for 20 minutes and washed twice with 2 ml of PBS. 1 mg / ml stock of DAPI was diluted 1/1000 in PBS, treated with 1 ml of cells, and 5 minutes later, 1 ml of PBS was added. The stained nuclei were observed at 400-fold using a fluorescence microscope and photographed. Experimental results showed that chromosome condensation did not occur, indicating that cell number reduction was not a result of Apoptosis (FIG. 1B).
다른 세포 증식 억제 원인을 알아보기 위해 세포분화 정도를 알 수 있는 Oil red O 염색법을 실시하였다. MH7A 세포를 6 well plate에 seeding하고 16시간 후 세포량이 60 % 정도일 때 PBS로 2번 워싱하고 Hemp 종자 추출물을 농도별로 섞은 RPMI 1640 배지 2 ml를 각 well에 처리하였다. 5 % CO2, 37 ℃ 조건에서 24시간 배양한 후 inverted microscopy에서 400배 확대한 세포사진을 찍고 PBS 2 ml로 2번 워싱하였다. 4 % paraformaldehyde 500 μl를 넣고 상온에서 1시간 고정하고 60 % Isoprophanol 1 ml로 2번 워싱하고 세포가 붙어있는 well을 완전히 말렸다. Oil red O 염색시약을 물과 6:4 비율로 섞어 0.2 μM filter로 filtering하여 working solution을 만들고 well에 500 μl씩 10분간 처리했다. Oil red O working solution을 제거하고 곧 바로 3차 증류수 1 ml로 5번 워싱 후 세포에 생긴 Oxdazed Lipid droplelet을 Inverted microscopy로 관찰하고 사진을 찍었다. 관찰 결과 Hemp seed oil 추출물이 MH7A 세포의 분화를 촉진한 것을 확인하였다 (도 1c). 이러한 결과는 hempseed oil 추출물이 류마티스 환자 활막 세포의 이상 증식 억제 효과를 가짐을 의미한다.Oil red O staining was performed to determine the extent of cell differentiation. MH7A cells were seeded in 6 well plates, and after 16 hours, when the cell volume was about 60%, the cells were washed twice with PBS and treated with 2 ml of RPMI 1640 medium containing Hemp seed extract at different concentrations. After culturing for 24 hours at 5% CO 2 , 37 ° C, 400 times magnified cell photographs were taken by inverted microscopy and washed twice with 2 ml of PBS. Add 500 μl of 4% paraformaldehyde, fix for 1 hour at room temperature, wash twice with 1 ml of 60% Isoprophanol, and dry wells with cells. Oil red O dyeing reagent was mixed with water at a 6: 4 ratio, filtered with a 0.2 μM filter to form a working solution, and then treated with 500 μl for 10 minutes. The oil red O working solution was removed and immediately washed five times with 1 ml of tertiary distilled water, followed by observing and photographing Oxdazed Lipid droplelet formed in the cells by Inverted microscopy. As a result, it was confirmed that Hemp seed oil extract promoted the differentiation of MH7A cells (FIG. 1C). These results indicate that hempseed oil extracts have an inhibitory effect on abnormal proliferation of synovial cells of rheumatoid patients.
실시예Example 2: 2: DHADHA 의 관절염에 대한 효과Effect on arthritis
Hemp seed oil의 지용성 유효성분인 EPA와 함께 Hemp seed oil의 유효성분으로 알려진 DHA(Sigma-Aldrich, St. Louis, MO)에 의한 활막세포 증식억제 효과를 조사하기 위해 MH7A에 DHA을 농도별로 처리하였다(도 2). DHA 처리 24시간 후 도 1a의 Cell counting Kit-8 방법을 통해 DHA의 농도가 증가할수록 MH7A 세포주의 증식이 억제됨을 확인하였다 (도 2a).To investigate the effect of DHA (Sigma-Aldrich, St. Louis, MO), known as the active ingredient of Hemp seed oil, along with EPA, the fat-soluble active ingredient of Hemp seed oil, DHA was treated by concentration in MH7A. (FIG. 2). After 24 hours of DHA treatment, it was confirmed that the growth of MH7A cell line was inhibited as the concentration of DHA increased through the Cell counting Kit-8 method of FIG.
DHA가 MH7A 세포에서 Apoptosis를 유도했는지 알아보기 위해 western blot을 통해 PARP cleavage 현상을 관찰하였다. MH7A 세포를 60 φ cell culture dish에 seeding하고 16시간 후 세포량이 60 % 정도일 때 PBS로 2번 워싱하고 DHA를 농도별로 섞은 RPMI 1640 배지 2 ml를 각 well에 처리하였다. 5 % CO2, 37 ℃ 조건에서 24시간 배양한 후 배양액과 세포를 원심분리를 통해 모은 후 Lysis solution (1% Protease inhibitor, Phosphatase inhibitor, Proteosome inhibitor가 포함된 RIPA buffer) 100 μl로 얼음에서 Lysis하였다. 4 ℃에서 15000 rpm으로 원심분리하여 soluble 단백질을 얻어 SDS-PAGE를 하였다. SDS gel에 전개된 단백질을 PVDF membrane에 transfer하고 anti-PARP와 anti-actin 항체를 이용하여 PARP cleavage 여부를 확인하였다 (도 2b). 실험 결과 DHA 100 μM에서 PARP cleaved form이 나타남을 확인하였다. 그리고 DAPI 염색법을 통해 DHA가 활막세포주의 염색체 응축을 유도하는 것을 확인하여 DHA의 활막세포주 이상증식 억제는 apoptosis에 의한 것임을 알았다 (도 2c).To determine whether DHA induced apoptosis in MH7A cells, PARP cleavage was observed through western blot. MH7A cells were seeded in a 60 φ cell culture dish, and after 16 hours, when the cell volume was about 60%, washed twice with PBS and treated with 2 ml of RPMI 1640 medium mixed with DHA in each well. After culturing for 24 hours at 5% CO 2 and 37 ℃, the culture medium and cells were collected by centrifugation and lysis on ice with 100 μl of Lysis solution (RIPA buffer containing 1% Protease inhibitor, Phosphatase inhibitor, Proteosome inhibitor). . SDS-PAGE was performed by obtaining a soluble protein by centrifugation at 15000 rpm at 4 ℃. The protein developed on the SDS gel was transferred to the PVDF membrane, and the presence of PARP cleavage was confirmed using anti-PARP and anti-actin antibodies (FIG. 2B). Experimental results showed that PARP cleaved form appeared in
이러한 결과들은 Hemp 종자의 지용성 추출물과 DHA가 류마티스 관절염 환자에서 유래된 활막세포의 이상증식을 억제하는 효과가 있으며 앞으로 류마티스 관절염 환자의 발병 방지 및 증상 완화를 위한 치료에 응용 가능한 결과이다.These results indicate that fat-soluble extracts of Hemp seeds and DHA have the effect of inhibiting the proliferation of synovial cells derived from rheumatoid arthritis patients, and are applicable to the treatment for preventing the onset and symptom relief of rheumatoid arthritis patients.
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JP2021061789A (en) * | 2019-10-15 | 2021-04-22 | サムライ金沢株式会社 | Food composition and feed composition |
KR102665874B1 (en) * | 2023-03-24 | 2024-05-10 | 가톨릭상지대학 산학협력단 | Feed composition for improving joint, skin and intestinal health of companion animals |
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EP3319446A4 (en) * | 2015-07-06 | 2019-07-03 | Cg- Bio Genomics, Inc. | Healthful supplements |
US11369655B2 (en) | 2015-07-06 | 2022-06-28 | Cg-Bio Genomics, Inc. | Healthful supplements |
JP2021061789A (en) * | 2019-10-15 | 2021-04-22 | サムライ金沢株式会社 | Food composition and feed composition |
KR102665874B1 (en) * | 2023-03-24 | 2024-05-10 | 가톨릭상지대학 산학협력단 | Feed composition for improving joint, skin and intestinal health of companion animals |
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