KR20180086935A - A composition for improving, preventing and treating arthritis comprising Stewartia koreana extract and Rhus verniciflua stokes extract - Google Patents
A composition for improving, preventing and treating arthritis comprising Stewartia koreana extract and Rhus verniciflua stokes extract Download PDFInfo
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- KR20180086935A KR20180086935A KR1020170011015A KR20170011015A KR20180086935A KR 20180086935 A KR20180086935 A KR 20180086935A KR 1020170011015 A KR1020170011015 A KR 1020170011015A KR 20170011015 A KR20170011015 A KR 20170011015A KR 20180086935 A KR20180086935 A KR 20180086935A
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- rhus verniciflua
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Abstract
Description
본 발명은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유함으로써, 관절염에 의해 발생할 수 있는 질환을 개선, 예방 또는 치료할 수 있는 조성물에 관한 것이다.The present invention relates to a composition capable of improving, preventing or treating a disease that may be caused by arthritis by containing an olfactory extract and an extract of Rhus verniciflua as an active ingredient.
관절염은 대표적인 퇴행성, 난치성 질환으로 전세계 인구의 12%, 60대 이상의 70%가 증상을 가지고 있으며, 우리나라 국민의 4~5%는 퇴행성관절염, 약 1%는 류마티스성 관절염 환자로 추정되며, 특히 55세 이상 노인의 80%가 퇴행성관절염을 가지고 있는 것으로 추정된다.Arthritis is a typical degenerative and intractable disease that affects 12% of the world's population and 70% of those in their 60s. Symptoms include arthritis, 4-5% of Koreans, and 1% of rheumatoid arthritis. It is estimated that 80% of the elderly aged people have degenerative arthritis.
상기 퇴행성관절염(degenerative arthritis)은 관절 연골이 닳아 없어지면서 국소적인 퇴행성 변화가 나타나는 질환으로서, 골관절염(osteoarthritis) 또는 골관절증이라고도 한다. 골관절염은 노령화와 밀접한 연관을 갖는 대표적인 퇴행성 질환으로, 전 인구의 10~15% 정도가 앓고 있으며, 특히 65세 이상의 고령인구 중 60~80% 정도가 골관절염을 앓고 있다.Degenerative arthritis is a disease in which degenerative arthritis of the joint cartilage is worn out, and is referred to as osteoarthritis or osteoarthritis. Osteoarthritis is a typical degenerative disease closely related to aging. It affects about 10 to 15% of the total population, and in particular, 60 to 80% of the elderly people aged 65 or older have osteoarthritis.
퇴행성관절염의 원인은 노화 현상이나 과다한 체중과 관계가 깊으며, 나이가 많아질수록 여성에게서 더 많이 그리고 더 심하게 나타난다. 초기 증상은 한개 또는 두개의 관절이 강직과 함께 쑤시는 듯한 통증이 나타나며, 장기화 되면 연골 하골의 경화, 관절 주변에 골의 과잉형성, 관절의 변형 등을 초래하게 된다.The cause of degenerative arthritis is related to aging phenomena and excessive weight, and the older the more and more the women appear. Early symptoms are painful with one or two joints tingling with stiffness, and when prolonged, hardening of cartilage bone, excessive bone formation around joints, and deformation of joints are caused.
퇴행성관절염은 일차성(원발성) 또는 이차성(속발성)으로 분류할 수 있다. 첫째, 일차성(원발성) 퇴행성관절염은 확실한 원인은 아직 밝혀져 있지 않지만, 나이, 성별, 유전적 요소, 비만증 등이 원인이라고 생각되고 있다. 일차성(원발성) 퇴행성관절염은 중년 이후 나이가 많을수록 발병 빈도가 높아지고, 여성에게서 더 많이 나타난다. 또한, 가족력과 관계가 있다. 비만증이 있는 경우는 정상인보다 약 2배 정도로 발생률이 높고, 이때는 주로 체중 부하 관절에 나타난다. 흔히 침범되는 관절은 체중 부하와 압박력을 받는 요추, 고관절, 슬관절, 그리고 발의 모지의 중족지 관절 등이다. 특히, 관절 내에서도 일정 부위, 즉 예를 들면 슬관절의 내측 관절이 잘 침범된다. Degenerative arthritis can be classified as primary (primary) or secondary (secondary). First, the definitive cause of primary (degenerative) arthritis is not yet known, but it is thought to be caused by age, sex, genetic factors, and obesity. Primary (primary) degenerative arthritis is more frequent in middle age and older, more frequent in women and more frequent in women. It is also related to family history. In the case of obesity, the incidence is about twice as high as that of the normal person, and this is mainly in the weight-bearing joint. Commonly involved joints are the lumbar, hip, knee, and hip joints of the moles of the foot that are under weight load and compression. Particularly, even within the joints, certain parts, for example, the inner joints of the knee joint, are well invaded.
여성의 경우는 수지의 원위지 관절(몸통에서 먼 손가락관절)과 제 1 수근중수 관절(손바닥의 관절)에 잘 침범되고, 남성의 경우는 고관절의 침범이 흔하다.In women, the lesion is well invaded in the distal joint of the resin (distal fingers in the torso) and in the first carpus metacarpophalangeal joint (in the palms of the palms). In males, the involvement of the hip joint is common.
둘째, 이차성(속발성) 퇴행성관절염은 관절 연골에 손상을 줄 수 있는 외상, 질병 및 기형이 모두 원인이 될 수 있다. 선천성 기형으로는 고관절 발육 부전이나 내반슬같이 관절의 정렬이 틀어진 경우이고, 화농성/결핵성 관절염후 관절 연골이 파괴된 경우는 무혈성 괴사, 심한 충격 또는 반복적인 가벼운 외상 등에서 퇴행성관절염이 속발되는 경우가 흔하다. 또한 말단 거대증이나 당뇨병 등 내분비이상이나, 통풍 등의 대사성 질환에서도 퇴행성관절염이 많이 나타난다.Second, secondary (secondary) degenerative arthritis can be caused by trauma, disease, and deformities that can damage the articular cartilage. Congenital anomalies are those in which joints are misaligned, such as hip dysplasia or varus, and arthritic arthritis is common in avascular necrosis, severe shock, or repeated minor trauma when arthritic cartilage is destroyed after pyogenic / tuberculous arthritis. Degenerative arthritis also occurs in metabolic diseases such as endocrine disorders such as terminal hypertension and diabetes, and gout.
관절연골의 퇴행성 변화의 원인은 아직 규명되어 있지는 않으며, 연골세포 수의 절대적 감소와 연골세포(chondrocytes)에서 일어나는 연골기질 합성과 분해의 불균형이 하나의 원인으로 알려져 있을 뿐이다. 따라서 현재까지 관절염은 근원 치료가 불가능하며, 지금까지 개발된 치료제들은 통증을 완화하고, 염증을 억제하며, 가능한 기능을 보전하는 방향으로 치료가 이루어지고 있다.The causes of degenerative changes in articular cartilage have not yet been elucidated, and there is only one known cause of the imbalance of cartilage matrix synthesis and degradation occurring in chondrocytes and the absolute decrease in cartilage cell number. So far, arthritis is not possible to cure the source, so far developed therapies to relieve pain, suppress inflammation, and is being treated in the direction of preserving the possible functions.
현재 임상적으로 사용되고 있는 퇴행성관절염의 치료로는, 약물치료제(진통제, 스테로이드제, 비스테로이드계 항염제 등)나 연골보호제(히알루론산, 글루코사민, 콘드로이틴 등)를 이용하거나 수술적 처치(관절경 수술, 경골 근위부 절골술, 관절 부분 치환술, 슬관절 전치환술 등)에 의한다. 그러나 약물치료제의 경우는 통증이나 염증반응 자체를 비특이적으로 완화시키는 효과만을 가지며, 연골보호제는 단지 연골세포에 영양을 공급해 주거나 충격을 완화시킴으로써 관절을 보호해 주는Treatment of degenerative arthritis, which is currently being used clinically, can be performed by using drugs (analgesics, steroids, nonsteroidal anti-inflammatory drugs) or cartilage protectants (hyaluronic acid, glucosamine, chondroitin, etc.) Tibial proximal osteotomy, joint replacement, and total knee arthroplasty). However, in the case of medicines, it only has the effect of nonspecifically relieving the pain or the inflammatory reaction itself, and the cartilage protecting agent only protects the joints by nourishing the cartilage cells,
역할을 할 뿐이다. 또한 스테로이드성 제제를 장기간 복용할 시 칼슘의 손실로 골다공증, 고혈압, 당뇨병 등을 초래할 수 있는 부작용이 있다. It only plays a role. In addition, there is a side effect that can cause osteoporosis, hypertension, diabetes and the like due to the loss of calcium when a steroid preparation is taken for a long time.
따라서 약물치료는 대부분의 경우 통증을 감소시키는 목적으로만 사용되고 있고, 영구적인 인공관절 치환술이 주를 이루고 있지만 근본적인 치료효과를 주는 약물이나 수술법은 현재까지 없는 실정이다.Therefore, drug therapy is mostly used only for the purpose of reducing pain, and although permanent artificial joint replacement is the mainstay, there is no drug or surgical method that has a fundamental therapeutic effect to date.
본 발명의 목적은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유함으로써, 관절염에 의해 발생할 수 있는 질환을 개선 또는 예방할 수 있는 식품 조성물을 제공하는데 있다.It is an object of the present invention to provide a food composition capable of improving or preventing a disease caused by arthritis by containing an olive tree extract and a Rhus verniciflua extract as an effective ingredient.
또한, 본 발명의 다른 목적은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유함으로써, 관절염에 의해 발생할 수 있는 질환을 예방 또는 치료할 수 있는 약학 조성물을 제공하는데 있다.Another object of the present invention is to provide a pharmaceutical composition which can prevent or treat a disease that may be caused by arthritis by containing an olive tree extract and Rhus verniciflua extract as an effective ingredient.
상기한 목적을 달성하기 위한 본 발명의 관절염을 개선 또는 예방할 수 있는 식품 조성물은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유할 수 있다.In order to achieve the above-mentioned object, the food composition of the present invention for improving or preventing arthritis may contain an extract of Lachander and Rhus verniciflua as an active ingredient.
상기 노각나무 추출물 및 옻나무 추출물은 1 : 2-10의 중량비로 혼합될 수 있다.The beech tree extract and Rhus verniciflua extract may be mixed at a weight ratio of 1: 2-10.
상기 노각나무 추출물은 노각나무 잎과 용매를 혼합하여 80 내지 120 ℃의 온도에서 1 내지 5시간 동안 추출한 것일 수 있다.The beech tree extract may be obtained by mixing the leaves of the olive leaves with a solvent and extracting the mixture at a temperature of 80 to 120 ° C for 1 to 5 hours.
상기 옻나무 추출물은 옻나무 목질부와 용매를 혼합하여 80 내지 120 ℃의 온도에서 1 내지 5시간 동안 추출한 것일 수 있다.The Rhus verniciflua extract may be obtained by mixing lacquer wood and a solvent and extracting the mixture at 80 to 120 ° C for 1 to 5 hours.
상기 용매는 물, 탄소수 1 내지 4의 저급알코올, 에틸렌글리콜, 에틸에테르 또는 이들의 혼합용매일 수 있다.The solvent may be water, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether or a mixture thereof.
상기 관절염은 퇴행성관절염(osteoarthritis), 류마티스관절염(Rhematoid arthritis), 강직성척추염(ankylosing spondylitis), 반응성관절염(reactive arthritis), 건선관절염, 전신성홍반성루프스(Systemic lupus erythematosus), 다발성근육염(polymyositis) 또는 류마티스다발근육통(polymyalgia rheumatica)일 수 있다.The arthritis may be selected from the group consisting of osteoarthritis, rheumatoid arthritis, ankylosing spondylitis, reactive arthritis, psoriatic arthritis, systemic lupus erythematosus, polymyositis or rheumatoid arthritis. It may be polymyalgia rheumatica.
또한, 상기한 다른 목적을 달성하기 위한 본 발명의 관절염을 예방 또는 치료할 수 있는 약학 조성물은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유할 수 있다.In addition, the pharmaceutical composition for preventing or treating arthritis of the present invention for achieving the above-mentioned other object may contain an extract of Lachung tree and Rhus verniciflua extract as an active ingredient.
상기 노각나무 추출물 및 옻나무 추출물은 1 : 2-10의 중량비로 혼합될 수 있다.The beech tree extract and Rhus verniciflua extract may be mixed at a weight ratio of 1: 2-10.
본 발명의 관절염의 개선, 예방 또는 치료용 조성물은 연골세포에 손상을 미치는 효소인 MMP-1, 3, 13의 유전자 발현을 효과적으로 억제하며, 손상된 연골을 재생시킬 수 있으므로 관절염 질환의 예방 및 치료에 효과적으로 사용될 수 있을 뿐만 아니라, 독성이 없으므로 식품의 형태로 섭취할 수 있다.The composition for improving, preventing or treating arthritis of the present invention effectively inhibits gene expression of MMP-1, 3, 13, which is an enzyme that damages cartilage cells, and regenerates damaged cartilage. Not only can it be used effectively, but it is toxic and can be ingested in the form of food.
도 1은 실시예 1, 비교예 1 및 2에 따라 제조된 추출물로 처리 시 연골세포의 회복에 미치는 영향을 나타낸 PCR이다.
도 2는 실시예 1, 비교예 1 및 2에 따라 제조된 추출물을 투여 시 손상된 연골의 재생에 미치는 영향을 조직 염색법으로 분석한 결과이다.FIG. 1 is a PCR showing the effect of the extract prepared according to Example 1 and Comparative Examples 1 and 2 on the recovery of chondrocytes when treated. FIG.
FIG. 2 shows the results of analysis of the effect of the extract prepared according to Example 1 and Comparative Examples 1 and 2 on the regeneration of injured cartilage by tissue staining.
본 발명은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유함으로써, 관절염에 의해 발생할 수 있는 질환을 개선, 예방 또는 치료할 수 있는 조성물에 관한 것이다.
The present invention relates to a composition capable of improving, preventing or treating a disease that may be caused by arthritis by containing an olfactory extract and an extract of Rhus verniciflua as an active ingredient.
이하, 본 발명을 상세하게 설명한다. Hereinafter, the present invention will be described in detail.
본 발명의 관절염의 개선, 예방 또는 치료용 조성물은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유한다. The composition for improving, preventing or treating arthritis according to the present invention contains an extract of Lachandrum tree and Rhus verniciflua extract as an active ingredient.
상기 노각나무 추출물 및 옻나무 추출물은 1 : 2-10의 중량비, 바람직하게는 1 : 4-7의 중량비, 더욱 바람직하게는 1 : 5의 중량비로 혼합된다. 노각나무 추출물을 기준으로 옻나무 추출물의 함량이 상기 하한치 미만인 경우에는 관절염의 개선, 예방 또는 치료 효능이 낮을 수 있으며, 상기 상한치 초과인 경우에는 관능성이 저하되고 관절염의 개선, 예방 또는 치료 효능이 낮을 수 있다. The beech tree extract and Rhus verniciflua extract are mixed at a weight ratio of 1: 2-10, preferably 1: 4-7, more preferably 1: 5. If the content of Rhus verniciflua extract is lower than the lower limit based on the extract of Lentinus edulis, the effect of improving, preventing or treating arthritis may be low. If the content is above the upper limit, the sensory properties are lowered and the efficacy, .
상기 노각나무 추출물 및 옻나무 추출물을 함께 사용하지 않고 단독으로 사용하는 경우에는 상기 두 추출물을 함께 사용하는 경우에 비하여 10 내지 70% 낮은 효능을 보인다. In case of using alone or in combination with the extracts of Raspberry and Rhus verniciflua, the extracts exhibit 10 to 70% lower efficacy than the case of using the extracts together.
상기 노각나무는 쌍떡잎식물 물레나물목 차나무과의 낙엽활엽 교목으로 한국, 일본 등지에 분포하며, 산 중턱 이상에서 자란다. 노각나무의 줄기껍질이나 잔가지를 말려서 다려 먹으면 간 질환 등에 효과적이라는 민간요법이 내려져 오고 있으나, 상기 노각나무와 잎 추출물에 대한 과학적인 연구는 아직까지 미흡한 실정이다. It is a deciduous broad-leaved arboreous tree, which is distributed in Korea and Japan. It grows in the middle of the mountainside. Although folk remedies have been suggested to be effective for the liver disease by drying the stem skin or twigs of the lobster tree, the scientific research on the lobster tree and leaf extracts is still insufficient.
상기 노각나무 추출물은 노각나무 잎을 건조시켜 분쇄한 후, 건조 중량의 1 내지 30배, 바람직하게는 5 내지 15배 부피의 용매에 투입하고, 80 내지 120 ℃의 온도에서 1 내지 5시간 동안 추출하는 것이 바람직하다.The beech tree extract is dried and pulverized, and then added to a solvent having a dry weight of 1 to 30 times, preferably 5 to 15 times the volume, and extracted at a temperature of 80 to 120 ° C for 1 to 5 hours .
또한, 상기 옻나무 추출물은 옻나무의 목질부인 칠목, 옻나무의 껍질인 칠피, 옻나무 수액의 건조물인 건칠 또는 이들 중 어느 하나 이상의 혼합물을 의미한다. 바람직하게는 우루시올과 같은 알레르기를 유발하는 독성물질 함량이 상대적으로 낮고, 유효성분 함량은 상대적으로 높은 옻나무 목재부인 칠목을 사용한다.In addition, the Rhus verniciflua extract refers to Rhus javanicum, Rhus javanica, Rhus javanicus, or Rhus javanicus; Preferably, the toxic substance causing allergy such as urushiol is relatively low and the active ingredient content is relatively high.
상기 옻나무 추출물은 옻나무를 분쇄한 후, 건조 중량의 1 내지 30배, 바람직하게는 5 내지 15배 부피의 용매에 투입하고, 80 내지 120 ℃의 온도에서 1 내지 5시간 동안 추출하는 것이 바람직하다.The Rhus verniciflua extract is preferably pulverized and then added to a solvent having a volume of 1 to 30 times, preferably 5 to 15 times its dry weight, and extracted at a temperature of 80 to 120 ° C for 1 to 5 hours.
상기 노각나무 및 옻나무를 추출하는데 사용되는 용매는 물, 탄소수 1 내지 4의 저급알코올, 에틸렌글리콜, 에틸에테르 또는 이들의 혼합용매인 것이 바람직하다.The solvent used for extracting the olfactory tree and Rhus verniciflua is preferably water, a lower alcohol having 1 to 4 carbon atoms, ethylene glycol, ethyl ether or a mixed solvent thereof.
추출 공정을 수행한 후에는 여과지로 여과하고 여액을 감압 진공농축기를 이용하여 20 내지 65 ℃의 온도에서 1 내지 10 시간 동안 농축시키는 것이 바람직하다. 더욱 바람직하게는 감압 농축한 다음에 농축물을 진공동결 건조 또는 열풍건조 또는 분사 방식에 의한 건조 등을 통하여 노각나무 추출물을 분말화하는 것이나, 이에 제한되는 것은 아니다. After the extraction process, it is filtered through a filter paper, and the filtrate is concentrated using a vacuum concentrator at a temperature of 20 to 65 ° C for 1 to 10 hours. More preferably, the concentrate is pulverized by vacuum lyophilization, hot-air drying or drying by spraying method after concentration under reduced pressure, but the present invention is not limited thereto.
상기 옻나무 추출물은 바람직하게는 옻나무에 존재하는 우루시올과 같은 알레르기를 유발하는 독성물질을 제거한 추출물이다. 옻나무의 독성물질을 제거하는 방법은 전통적인 옻나무의 독성물질 제거방법인 자연건조 또는 초법(抄法; 볶음)을 이용할 수 있고, 상기 전통적인 옻나무 독성물질 제거방법을 촉진하기 위해 산화제, 키토산 등의 첨가제를 넣거나 분쇄한 후 건조 또는 초법을 수행하거나, 옻나무를 과초산과 혼합하거나, 옻나무를 전자이온화에너지로 처리하거나, 옻나무를 마이크로웨이브로 처리하거나, 옻나무를 버섯 균주, 효모 등의 미생물로 발효시키는 방법 어느 것이나 제한 없이 이용할 수 있다.The Rhus verniciflua extract is preferably an extract obtained by removing toxic substances such as urushiol present in Rhus verniciflua causing allergy. In order to remove the toxic substance of the lacquer tree, it is possible to use a natural drying method or a so-called roast method, which is a conventional method for removing toxic substances from the lacquer tree, and an additive such as an oxidizing agent or chitosan A method in which the lacquer is mixed with peracetic acid, the lacquer is treated with electron ionization energy, the lacquer is treated with microwave, or the lacquer is fermented with microorganisms such as mushroom strain or yeast It can be used without limitation.
예를 들어 옻나무 목재부에서 우루시올과 같은 알레르기를 유발하는 독성물질을 제거하기 위하여, 옻나무의 목재부를 절단하여 그늘에서 1 주일 이상, 바람직하게는 1 개월 이상, 더욱 바람직하게는 3 개월 이상 건조한 후, 파쇄시키거나 그대로 사용할 수 있다. 상기 건조 기간의 상한치는 별도로 설정할 필요없고, 1년 또는 수년 이상 건조된 것을 사용할 수도 있다. 또한 옻나무의 목재부를 건조시키지 않고 목재칩이나 톱밥으로 제조하여 사용할 수도 있다.For example, in order to remove toxic substances that cause allergies such as urushiol in the lacquer wood part, the wood part of the lacquer tree is cut and dried in the shade for 1 week or more, preferably 1 month or more, more preferably 3 months or more, It can be crushed or used as is. The upper limit of the drying period is not required to be set separately, and it may be dried for one year or several years or more. Also, the wood part of the lacquer tree can be used as a wood chip or sawdust without being dried.
본 명세서에서 노각나무 및 옻나무를 언급하면서 사용되는 용어 '추출물'은 추출용매를 처리하여 얻은 조추출물뿐만 아니라 노각나무 추출물 및 옻나무 추출물의 가공물도 포함한다. 예를 들어, 노각나무 추출물 및 옻나무 추출물은 감압 증류 및 동결 건조 또는 분무 건조 등과 같은 추가적인 과정에 의해 분말 상태로 제조될 수 있다.The term 'extract' used in the present specification to refer to the olive tree and Rhus verniciflua includes not only crude extract obtained by treating the extracting solvent but also the products of olive tree extract and Rhus verniciflua extract. For example, olive tree extract and Rhus verniciflua extract can be prepared in powder form by further processes such as vacuum distillation and lyophilization or spray drying.
또한, 본 발명의 노각나무 추출물 및 옻나무 추출물은 광의로는 노각나무 및 옻나무를 동물에게 투여할 수 있도록 제형화된 노각나무 및 옻나무 가공물, 예컨대, 노각나무 및 옻나무 분말도 포함하는 의미를 갖는다. 비록 본 발명에서 노각나무 추출물 및 옻나무 추출물로 실험을 진행하긴 하였으나, 노각나무 및 옻나무 가공물과 같은 형태로도 목적하는 효과를 달성할 수 있음은 당업자라면 예상가능할 것이다.In addition, the extracts of the olive tree and Rhus verniciflua according to the present invention have broad meaning as including lenticel and lacquer products formulated to be administered to animals in the form of lobsters and lacquer trees, such as lobsters and Rhus verniciflua. Although the present invention has been carried out with the extracts of Lachlan and Rhus verniciflua, it is expected that those skilled in the art will be able to achieve the desired effects in the form of lobsters and lacquer products.
한편, 본 명세서에서 용어 '유효성분으로 함유하는'이란 노각나무 추출물 및 옻나무 추출물의 효능 또는 활성을 달성하는 데 충분한 양을 포함하는 것을 의미한다. 일예로, 상기 노각나무 추출물 및 옻나무 추출물은 10 내지 1500 ㎍/㎖, 바람직하게는 100 내지 1000 ㎍/㎖의 농도로 사용된다. 노각나무 추출물 및 옻나무 추출물은 천연물로서 과량 투여하여도 인체에 부작용이 없으므로 본 발명의 조성물 내에 포함되는 노각나무 추출물 및 옻나무 추출물의 양적 상한은 당업자가 적절한 범위 내에서 선택하여 실시할 수 있다.
In the present specification, the term " comprising as an active ingredient " is meant to include an amount sufficient to achieve the potency or activity of the olive tree extract and the Rhus verniciflua extract. For example, the olive tree extract and Rhus verniciflua extract are used at a concentration of 10 to 1500 μg / ml, preferably 100 to 1000 μg / ml. Since the lumber extract and Rhus verniciflua extract have no adverse effects on the human body even when they are administered in an excess amount as a natural product, the quantitative upper limit of the extract of Rhus verniciflua and Rhus verniciflua extract contained in the composition of the present invention can be selected by a person skilled in the art within a suitable range.
본 발명의 약제학적 조성물은 상기 유효 성분 이외에 약제학적으로 적합하고 생리학적으로 허용되는 보조제를 사용하여 제조될 수 있으며, 상기 보조제로는 부형제, 붕해제, 감미제, 결합제, 피복제, 팽창제, 윤활제, 활택제 또는 향미제 등을 사용할 수 있다.The pharmaceutical composition of the present invention can be prepared by using pharmaceutically acceptable and physiologically acceptable adjuvants in addition to the above-mentioned active ingredients. Examples of the adjuvants include excipients, disintegrants, sweeteners, binders, coating agents, swelling agents, lubricants, A lubricant or a flavoring agent can be used.
상기 약제학적 조성물은 투여를 위해서 상기 기재한 유효 성분 이외에 추가로 약제학적으로 허용 가능한 담체를 1종 이상 포함하여 약제학적 조성물로 바람직하게 제제화할 수 있다.The pharmaceutical composition may be formulated into a pharmaceutical composition containing at least one pharmaceutically acceptable carrier in addition to the above-described active ingredients for administration.
상기 약제학적 조성물의 제제 형태는 과립제, 산제, 정제, 피복정, 캡슐제, 좌제, 액제, 시럽, 즙, 현탁제, 유제, 점적제 또는 주사 가능한 액제 등이 될 수 있다. 예를 들어, 정제 또는 캡슐제의 형태로의 제제화를 위해, 유효 성분은 에탄올, 글리세롤, 물 등과 같은 경구, 무독성의 약제학적으로 허용 가능한 불활성 담체와 결합될 수 있다. 또한, 원하거나 필요한 경우, 적합한 결합제, 윤활제, 붕해제 및 발색제 또한 혼합물로 포함될 수 있다. 적합한 결합제는 이에 제한되는 것은 아니나, 녹말, 젤라틴, 글루코스 또는 베타-락토오스와 같은 천연 당, 옥수수 감미제, 아카시아, 트래커캔스 또는 소듐올레이트와 같은 천연 및 합성 검, 소듐 스테아레이트, 마그네슘 스테아레이트, 소듐 벤조에이트, 소듐 아세테이트, 소듐 클로라이드 등을 포함한다. 붕해제는 이에 제한되는 것은 아니나, 녹말, 메틸 셀룰로스, 아가, 벤토니트, 잔탄 검 등을 포함한다.The pharmaceutical composition may be in the form of granules, powders, tablets, coated tablets, capsules, suppositories, liquids, syrups, juices, suspensions, emulsions, drops or injectable solutions. For example, for formulation into tablets or capsules, the active ingredient may be combined with an oral, non-toxic pharmaceutically acceptable inert carrier such as ethanol, glycerol, water, and the like. Also, if desired or necessary, suitable binders, lubricants, disintegrants and coloring agents may also be included as a mixture. Suitable binders include, but are not limited to, natural sugars such as starch, gelatin, glucose or beta-lactose, natural and synthetic gums such as corn sweeteners, acacia, tracker candles or sodium oleate, sodium stearate, magnesium stearate, sodium Benzoate, sodium acetate, sodium chloride, and the like. Disintegrants include, but are not limited to, starch, methyl cellulose, agar, bentonite, xanthan gum and the like.
액상 용액으로 제제화되는 조성물에 있어서 허용 가능한 약제학적 담체로는, 멸균 및 생체에 적합한 것으로서, 식염수, 멸균수, 링거액, 완충 식염수, 알부민 주사용액, 덱스트로즈 용액, 말토 덱스트린 용액, 글리세롤, 에탄올 및 이들 성분 중 1 성분 이상을 혼합하여 사용할 수 있으며, 필요에 따라 항산화제, 완충액, 정균제 등 다른 통상의 첨가제를 첨가할 수 있다. 또한 희석제, 분산제, 계면활성제, 결합제 및 윤활제를 부가적으로 첨가하여 수용액, 현탁액, 유탁액 등과 같은 주사용 제형, 환약, 캡슐, 과립 또는 정제로 제제화할 수 있다.Acceptable pharmaceutical carriers for compositions that are formulated into a liquid solution include sterile solutions suitable for the living body such as saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, One or more of these components may be mixed and used. If necessary, other conventional additives such as an antioxidant, a buffer, and a bacteriostatic agent may be added. In addition, diluents, dispersants, surfactants, binders, and lubricants may be additionally added to formulate into injectable solutions, pills, capsules, granules or tablets such as aqueous solutions, suspensions, emulsions and the like.
더 나아가 해당분야의 적절한 방법으로 Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA에 개시되어 있는 방법을 이용하여 각 질환에 따라 또는 성분에 따라 바람직하게 제제화할 수 있다.Further, it can be suitably formulated according to each disease or ingredient, using the method disclosed in Remington's Pharmaceutical Science, Mack Publishing Company, Easton PA as an appropriate method in the field.
본 발명의 약제학적 조성물은 경구 또는 비경구로 투여할 수 있고, 비경구 투여인 경우에는 정맥내 주입, 피하 주입, 근육 주입, 복강 주입, 경피 투여 등으로 투여할 수 있으며, 바람직하게는 경구 투여이다.The pharmaceutical composition of the present invention can be administered orally or parenterally. In the case of parenteral administration, the composition can be administered by intravenous injection, subcutaneous injection, muscle injection, intraperitoneal injection, transdermal administration, etc., .
본 발명의 약제학적 조성물의 적합한 투여량은 제제화 방법, 투여 방식, 환자의 연령, 체중, 성, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사는 소망하는 치료 또는 예방에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다. 본 발명의 바람직한 구현예에 따르면, 본 발명의 약제학적 조성물의 1일 투여량은 0.001-10 g/㎏이다.The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, age, body weight, sex, pathological condition, food, administration time, administration route, excretion rate and responsiveness of the patient, Usually, a skilled physician can readily determine and prescribe dosages effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 0.001-10 g / kg.
본 발명의 약제학적 조성물은 약제학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기 내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액 형태이거나 엑스제, 분말제, 과립제, 정제 또는 캅셀제 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention can be prepared in unit dose form by formulating it with a pharmaceutically acceptable carrier and / or excipient or can be manufactured by inserting it into a multi-dose container. The formulations may be in the form of solutions, suspensions or emulsions in oils or aqueous media, or in the form of excipients, powders, granules, tablets or capsules, and may additionally contain dispersing or stabilizing agents.
또한, 본 발명은 노각나무 추출물 및 옻나무 추출물을 유효성분으로 함유하는 관절염의 개선, 예방 또는 치료용 식품 조성물을 제공한다.In addition, the present invention provides a food composition for improving, preventing or treating arthritis containing an olfactory extract and Rhus verniciflua extract as an active ingredient.
본 발명에 따른 식품 조성물은 상기 약제학적 조성물과 동일한 방식으로 제제화되어 기능성 식품으로 이용하거나, 각종 식품에 첨가할 수 있다. 본 발명의 조성물을 첨가할 수 있는 식품으로는 예를 들어, 음료류, 알코올 음료류, 과자류, 다이어트바, 유제품, 육류, 초코렛, 피자, 라면, 기타 면류, 껌류, 아이스크림류, 비타민 복합제, 건강보조식품류 등이 있다.The food composition according to the present invention can be formulated in the same manner as the above pharmaceutical composition and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, alcoholic beverages, confectioneries, diet bars, dairy products, meat, chocolates, pizza, ram noodles, other noodles, gums, ice cream, .
본 발명의 식품 조성물은 유효성분으로서 노각나무 추출물 및 옻나무 추출물뿐만 아니라, 식품 제조 시에 통상적으로 첨가되는 성분을 포함할 수 있으며, 예를 들어, 단백질, 탄수화물, 지방, 영양소, 조미제 및 향미제를 포함한다. 상술한 탄수화물의 예는 모노사카라이드, 예를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 슈크로스, 올리고당 등; 및 폴리사카라이드, 예를 들어 덱스트린, 사이클로덱스트린 등과 같은 통상적인 당 및 자일리톨, 소르비톨, 에리트리톨 등의 당알콜이다. 향미제로서 천연 향미제 [타우마틴, 스테비아 추출물 (예를 들어 레바우디오시드 A, 글리시르히진 등]) 및 합성 향미제(사카린, 아스파르탐 등)를 사용할 수 있다. 예컨대, 본 발명의 식품 조성물이 드링크제와 음료류로 제조되는 경우에는 본 발명의 노각나무 추출물 및 옻나무 추출물 이외에 구연산, 액상과당, 설탕, 포도당, 초산, 사과산, 과즙, 및 각종 식물 추출액 등을 추가로 포함시킬 수 있다.The food composition of the present invention may contain, as an active ingredient, a component such as a Lentinus edulis extract and a Rhus verniciflua extract as well as a component that is ordinarily added at the time of food production. Examples thereof include protein, carbohydrate, fat, nutrients, . Examples of the above-mentioned carbohydrates are monosaccharides such as glucose, fructose, and the like; Disaccharides such as maltose, sucrose, oligosaccharides and the like; And polysaccharides such as dextrin, cyclodextrin and the like, and sugar alcohols such as xylitol, sorbitol and erythritol. Natural flavorings such as tau martin and stevia extract (e.g., rebaudioside A and glycyrrhizin) and synthetic flavorings (saccharine, aspartame, etc.) can be used as flavorings. For example, when the food composition of the present invention is prepared from a drink and a beverage, it may further contain citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, juice, various plant extracts and the like in addition to the olive tree extract and Rhus verniciflua extract of the present invention .
본 발명은 상기 노각나무 추출물 및 옻나무 추출물을 유효성분으로 포함하는 관절염의 개선, 예방 또는 치료용 식품 조성물을 포함하는 건강기능식품을 제공한다. 건강기능식품이란, 노각나무 추출물 및 옻나무 추출물을 음료, 차류, 향신료, 껌, 과자류 등의 식품소재에 첨가하거나, 캡슐화, 분말화, 현탁액 등으로 제조한 식품으로, 이를 섭취할 경우 건강상 특정한 효과를 가져오는 것을 의미하나, 일반 약품과는 달리 식품을 원료로 하여 약품의 장기 복용시 발생할 수 있는 부작용 등이 없는 장점이 있다. 이와 같이 하여 얻어지는 본 발명의 건강기능식품은, 일상적으로 섭취하는 것이 가능하기 때문에 매우 유용하다. 이와 같은 건강기능식품에 있어서의 노각나무 추출물 및 옻나무 추출물의 첨가량은, 대상인 건강기능식품의 종류에 따라 달라 일률적으로 규정할 수 없지만, 식품 본래의 맛을 손상시키지 않는 범위에서 첨가하면 되며, 대상 식품에 대하여 통상 0.01 내지 50 중량%, 바람직하기로는 0.1 내지 20 중량%의 범위이다. 또한, 환제, 과립제, 정제 또는 캡슐제 형태의 건강기능식품의 경우에는 통상 0.1 내지 100 중량% 바람직하기로는 0.5 내지 80 중량%의 범위에서 첨가하면 된다. 한 구체예에서, 본 발명의 건강기능식품은 환제, 정제, 캡슐제 또는 음료의 형태일 수 있다.The present invention provides a health functional food comprising a food composition for improving, preventing or treating arthritis, which comprises the extract of Raspberry and Rhus verniciflua as an active ingredient. The health functional food is a food prepared by adding a lobster extract and Rhus verniciflua extract to a food material such as a beverage, a tea, a spice, a gum or a confectionery, or encapsulating, powdering or suspending, However, unlike general medicine, there is an advantage that there is no side effect that may occur when a food is used as a raw material for a long period of taking the medicine. The health functional food of the present invention thus obtained is very useful because it can be ingested routinely. The amount of the extract of Lentinus edulis extract and Rhus verniciflua extract in such a health functional food can not be uniformly determined depending on the kind of the health functional food to which it is added but may be added within a range that does not deteriorate the original taste of the food, Is usually in the range of 0.01 to 50% by weight, preferably 0.1 to 20% by weight. In the case of health functional foods in the form of pills, granules, tablets or capsules, they may be added usually in the range of 0.1 to 100% by weight, preferably 0.5 to 80% by weight. In one embodiment, the health functional food of the present invention may be in the form of a pill, tablet, capsule or beverage.
또한, 본 발명은 관절염의 개선, 예방 또는 치료용 의약 또는 식품의 제조를 위한 노각나무 추출물 및 옻나무 추출물의 용도를 제공한다. 상기한 바와 같이 노각나무 추출물 및 옻나무 추출물은 관절염의 개선, 예방 또는 치료를 위한 용도로 이용될 수 있다.In addition, the present invention provides the use of lachryme extract and Rhus verniciflua extract for the manufacture of a medicament or food for improving, preventing or treating arthritis. As described above, olive tree extract and Rhus verniciflua extract can be used for improving, preventing or treating arthritis.
또한, 본 발명은 포유동물에게 유효량의 노각나무 추출물 및 옻나무 추출물을 투여하는 것을 포함하는 관절염의 개선, 예방 또는 치료 방법을 제공한다.The present invention also provides a method of improving, preventing, or treating arthritis, comprising administering to a mammal an effective amount of Lentinus edulis extract and Rhus verniciflua extract.
여기에서 사용된 용어 "포유동물"은 치료, 관찰 또는 실험의 대상인 포유동물을 말하며, 바람직하게는 인간을 말한다.The term "mammal " as used herein refers to a mammal that is the subject of treatment, observation or experimentation, preferably a human.
여기에서 사용된 용어 "유효량"은 연구자, 수의사, 의사 또는 기타 임상의에 의해 생각되는 조직계, 동물 또는 인간에서 생물학적 또는 의학적 반응을 유도하는 유효 성분 또는 약학적 조성물의 양을 의미하는 것으로, 이는 해당 질환 또는 장애의 증상의 완화를 유도하는 양을 포함한다. 본 발명의 유효 성분에 대한 유효량 및 투여횟수는 원하는 효과에 따라 변화될 수 있다. 그러므로, 투여될 최적의 투여량은 당업자에 의해 쉽게 결정될 수 있으며, 질환의 종류, 질환의 중증도, 조성물에 함유된 유효성분 및 다른 성분의 함량, 제형의 종류, 및 환자의 연령, 체중, 일반 건강 상태, 성별 및 식이, 투여 시간, 투여 경로 및 조성물의 분비율, 치료기간, 동시 사용되는 약물을 비롯한 다양한 인자에 따라 조절될 수 있다. 본 발명의 예방, 치료 또는 개선 방법에 있어서, 성인의 경우, 노각나무 추출물 및 옻나무 추출물을 1일 1회 내지 수회 투여시, 0.001 g/kg 내지 10 g/kg의 용량으로 투여하는 것이 바람직하다.As used herein, the term "effective amount" refers to the amount of active ingredient or pharmaceutical composition that elicits a biological or medical response in a tissue system, animal, or human, as contemplated by a researcher, veterinarian, physician or other clinician, ≪ / RTI > inducing a reduction of the symptoms of the disease or disorder. The effective amount and the administration frequency for the active ingredient of the present invention can be changed according to the desired effect. Thus, the optimal dosage to be administered can be readily determined by those skilled in the art and will vary with the nature of the disease, the severity of the disease, the amount of active and other ingredients contained in the composition, the type of formulation, and the age, The age, body weight, sex, diet, time of administration, route of administration and fraction of the composition, duration of treatment, concurrent medication, and the like. In the prevention, treatment, or improvement of the present invention, it is preferable to administer the extracts of Lachmini and Rhus verniciflua in an amount of 0.001 g / kg to 10 g / kg once or several times a day.
본 발명의 치료방법에서 노각나무 추출물 및 옻나무 추출물을 유효 성분으로 포함하는 조성물은 경구, 직장, 정맥내, 동맥내, 복강내, 근육내, 흉골내, 경피, 국소, 안구내 또는 피내 경로를 통해 통상적인 방식으로 투여할 수 있다.
In the treatment method of the present invention, the composition comprising an extract of Lachand L. and extract of Rhus verniciflua as an active ingredient can be administered orally, rectally, intravenously, intraarterially, intraperitoneally, intramuscularly, intrasternally, transdermally, topically, And can be administered in a conventional manner.
이하, 본 발명의 이해를 돕기 위하여 바람직한 실시예를 제시하나, 하기 실시예는 본 발명을 예시하는 것일 뿐 본 발명의 범주 및 기술사상 범위 내에서 다양한 변경 및 수정이 가능함은 당업자에게 있어서 명백한 것이며, 이러한 변형 및 수정이 첨부된 특허청구범위에 속하는 것도 당연한 것이다.It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the present invention. Such variations and modifications are intended to be within the scope of the appended claims.
실시예 1. 노각나무 잎 추출물 : 옻나무 추출물 = 1 : 5의 중량비Example 1: Weight ratio of extract of Leaf Tree Leaf: Rhus verniciflua = 1: 5
노각나무 잎 추출물Leaf Leaf Extract
노각나무(Stewartia pseudocamellia Maxim.) 잎은 2016년 7월 전라남도 광양에서 채집하였으며 정확히 감정한 후 음건하여 사용하였다. 상기 노각나무의 건조된 잎에 10배 중량의 주정을 첨가하여 100 ℃의 온도에서 3시간 동안 추출하고 여과 후 감압 농축하여 노각나무 잎 추출물을 수득하였다.Leaves were collected in Gwangyang, Jeollanam - do in July 2016, and they were used for shade evaluation. The dried leaf of the beech tree was added with 10 times the weight of the alcohol, and the mixture was extracted at a temperature of 100 ° C for 3 hours, filtered and concentrated under reduced pressure to obtain a Leaf Leaf Extract.
옻나무 추출물Rhus verniciflua extract
강원도 원주시 일대에서 채집한 옻나무의 목질부를 충분히 자연건조시킨 후The woody part of the lacquer tree collected in Wonju city, Gangwon province was sufficiently dried naturally
목재칩 제조기에 약 11X1X0.2 cm 크기로 파쇄하였다. 상기 옻나무 목재칩에 10배 중량의 물을 첨가하여 100 ℃ 이상에서 4시간 동안 열수 추출하고 여과 후 감압 농축하여 15 브릭스의 옻나무 열수추출물을 수득하였다.The wood chips were crushed to about 11X1X0.2 cm in size. 10 times by weight water was added to the lacquer wood chips, and the mixture was subjected to hot water extraction at 100 DEG C or higher for 4 hours, filtered, and then concentrated under reduced pressure to obtain a 15-brix Rhus verniciflua extract.
혼합 추출물Mixed extract
상기 제조된 노각나무 잎 추출물과 상기 제조된 옻나무 추출물을 1 : 5의 중량비로 혼합하였다.
The prepared olive leaf extract and Rhus verniciflua extract were mixed at a weight ratio of 1: 5.
비교예 1. 노각나무 잎 추출물Comparative Example 1. Leaf leaf extract
노각나무(Stewartia pseudocamellia Maxim.) 잎은 2016년 7월 전라남도 광양에서 채집하였으며 정확히 감정한 후 음건하여 사용하였다. 상기 노각나무의 건조된 잎 14 kg을 주정 140 L로 100 ℃의 온도에서 3시간 동안 추출하고 여과 후 감압 농축하여 노각나무 잎 추출물을 수득하였다.
Leaves were collected in Gwangyang, Jeollanam - do in July 2016, and they were used for shade evaluation. 14 kg of dried leaves of the beech tree were extracted with 140 liters of water at a temperature of 100 ° C for 3 hours, filtered and concentrated under reduced pressure to obtain a Leaf Leaf Extract.
비교예 2. 옻나무 추출물Comparative Example 2. Rhus verniciflua extract
강원도 원주시 일대에서 채집한 옻나무의 목질부를 충분히 자연건조시킨 후 목재칩 제조기에 약 11X1X0.2 cm 크기로 파쇄하였다. 상기 옻나무 목재칩에 10배 중량의 물을 첨가하여 100 ℃ 이상에서 4시간 동안 열수 추출하고 여과 후 감압 농축하여 15 브릭스의 옻나무 열수추출물을 수득하였다.
The woody part of the lacquer tree collected from Wonju city, Gangwon province was thoroughly naturally dried and then crushed to a size of about 11 × 1 × 0.2 cm on a wood chip maker. 10 times by weight water was added to the lacquer wood chips, and the mixture was subjected to hot water extraction at 100 DEG C or higher for 4 hours, filtered, and then concentrated under reduced pressure to obtain a 15-brix Rhus verniciflua extract.
<시험예><Test Example>
시험예 1.Test Example 1 세포 생존능 측정Cell viability measurement
10 ul의 HUVECs(혈관내피세포; human vascular endothelial cell, 아메리칸 타입 컬쳐 컬렉션(Manassas, VA, USA))을 이용하여 신생혈관 형성을 유도하지 않고, 실시예 및 비교예의 추출물을 농도 의존적으로 처리한 후 세포사멸 경향을 WST-1 분석법으로 측정하였다.The extracts of Examples and Comparative Examples were treated in a concentration-dependent manner without inducing angiogenesis using 10 ul of HUVECs (human vascular endothelial cell, American Type Culture Collection (Manassas, Va., USA) The cell death tendency was measured by WST-1 assay.
위 표 1에 나타낸 바와 같이, 본 발명의 실시예 1 내지 3에 따라 제조된 추출물 및 비교예 1 내지 2에 따라 제조된 추출물 모두 세포 생존능이 우수하므로 세포 독성이 낮은 것을 확인하였다.
As shown in Table 1, the extracts prepared according to Examples 1 to 3 of the present invention and the extracts prepared according to Comparative Examples 1 and 2 were found to have low cytotoxicity due to their excellent cell viability.
시험예Test Example 2. 염증 자극에 의한 사람 연골세포의 손상에 관여하는 인자들의 유전자 발현에 혼합 추출물이 미치는 영향 분석 2. Effects of mixed extracts on gene expression of factors involved in damage to human chondrocytes by inflammatory stimulation
Human chondrocyte를 10% FBS가 포함된 CGM으로 suspend하여 6 well cell culture plate에 1X106 cells/well로 분주한 뒤 80% 이상 자랄 때까지 5% CO2가 공급되는 37 ℃ 배양기에서 배양하였다. 대표적인 염증 유도 사이토카인인 TNF-α(tumor necrosis factor-alpha)가 10 ng/mL로 포함된 0% CBM을 사용하여 실시예 1, 비교예 1 및 2의 추출물을 각각 0, 20, 100 μg/mL의 농도로 희석하여 준비하고, 6 well 플레이트를 1 mL PBS로 2번 세척한 뒤, 준비된 각 추출물 시료를 2 mL씩 처리하고 22시간 동안 배양하였다.The human chondrocyte was suspended in CGM containing 10% FBS, and the cells were plated on a 6-well cell culture plate at 1 × 10 6 cells / well. Then, the cells were cultured in a 37 ° C. incubator in which 5% The extracts of Example 1 and Comparative Examples 1 and 2 were supplemented with 0, 20 and 100 μg / ml of 0% CBM containing 10 ng / ml of tumor necrosis factor-alpha (TNF-α) mL, and the 6-well plate was washed twice with 1 mL of PBS. Each of the prepared extracts was treated with 2 mL each and cultured for 22 hours.
22시간 후 플레이트를 석션한 뒤, RNA-Bee를 각 well에 1 mL 씩 첨가하여 RNA를 분리하고, 분리된 RNA 5 μg을 사용하여 cDNA를 합성한 후, 합성된 cDNA를 템플레이트로 사용하고 β-actin 유전자를 internal control로 사용하여, 연골세포에 손상을 미치는 효소인 MMPs(matrix metalloproteinases)중 주요 인자로 보고된 MMP-1, 3, 13의 유전자 발현을 PCR로 확인하였다.After 22 hours, the plate was suctioned, RNA was separated by adding 1 mL of RNA-Bee to each well, and 5 μg of the separated RNA was used to synthesize cDNA. The synthesized cDNA was used as a template, Using the actin gene as an internal control, the expression of MMP-1, 3, and 13 genes, which are reported as key factors in MMPs (matrix metalloproteinases), an enzyme that damages cartilage cells, was confirmed by PCR.
단, PCR 실험에 사용된 각 유전자의 primer sequence와 PCR 조건은 하기 표 2에서 보는 바와 같은 조건으로 실시하였다.However, the primer sequences and PCR conditions of each gene used in the PCR experiment were performed as shown in Table 2 below.
R: ttg gac tca cac cat gtg ttF: ctg ttc agg gac aga atg tg
R: ttg gac tca cac cat gtg tt
R: gaa tgt gag tgg agt cac ctF: tgc gtg gca gtt tgc tca gc
R: gaa tgt gag tgg agt cac ct
R: atc aaa tgg gtagaa gtc gcF: ggc tcc gag aaa tgc agt ct
R: atc aaa tgg gtagaa gtc gc
R: cac tgt gtt ggc gta cag gtF: tga gac ctt caa cac ccc ag
R: cac tgt gtt ggc gta cag gt
도 1은 실시예 1, 비교예 1 및 2에 따라 제조된 추출물로 처리 시 연골세포의 회복에 미치는 영향을 나타낸 PCR이다.FIG. 1 is a PCR showing the effect of the extract prepared according to Example 1 and Comparative Examples 1 and 2 on the recovery of chondrocytes when treated. FIG.
연골세포에 손상을 미치는 효소인 MMP-1, 3, 13의 유전자 발현을 억제하는 것을 관찰하였으며, 상기 MMP-1, MMP-3 및 MMP-13은 MMP(matrix metalloproteinase)계열로, 인체의 연골세포에 손상을 미치는 효소로서 상기 효소가 발현됨에 따라 연골세포에 손상을 유도하는 역할을 한다.MMP-1, MMP-3, and MMP-13 were identified as MMP (matrix metalloproteinase) -based cells, and the human cartilage cells Which acts to induce damage to chondrocytes as the enzyme is expressed.
즉, 상기 MMP의 활성을 저해하여 연골세포의 손상을 억제함으로써 관절 내 염증을 억제하여 관절염을 예방 또는 개선시키는 것이 중요하다.That is, it is important to prevent or ameliorate arthritis by inhibiting inflammation in the joint by inhibiting the activity of the MMP to inhibit damage of cartilage cells.
도 1에 도시된 바와 같이, 실시예 1의 혼합 추출물은 염증 유도 물질인 TNF-α에 의해 유도되는 MMP-1, 3, 13의 발현을 우수하게 저해시키는 것을 확인하였다. 반면, 비교예 1 및 비교예 2에서 제조된 노각 추출물 단독 및 옻 추출물 단독은 염증 유도 물질인 TNF-α에 의해 유도되는 MMP-1, 3, 13의 발현을 저해시키지 못하는 것을 확인하였다. As shown in FIG. 1, it was confirmed that the mixed extract of Example 1 excellently inhibited the expression of MMP-1, 3, 13 induced by TNF-α, an inflammation inducer. On the other hand, it was confirmed that the extracts of Lambskin extract alone and raspberry extract alone prepared in Comparative Example 1 and Comparative Example 2 did not inhibit the expression of MMP-1, 3 and 13 induced by TNF-α, an inflammation inducer.
이러한 결과는 실시예 1의 혼합 추출물이 관절에서 염증발생 후 연골의 손상 방지와 손상된 연골의 재생에 효과적인 물질로 사용될 수 있음을 의미한다.
These results indicate that the mixed extract of Example 1 can be used as an effective material for prevention of cartilage damage and regeneration of injured cartilage after inflammation in joints.
시험예 3. 자극에 의해 연골이 손상된 동물모델(OD model)을 활용한 혼합 추출물의 연골 재생 활성 분석Test Example 3. Analysis of cartilage regeneration activity of mixed extracts using an animal model (OD model) in which cartilage was damaged by stimulation
Human chondrocyte를 이용한 실시예 1, 비교예 1 및 2 추출물의 세포독성 분석 결과 최고 처리 농도인 1000 μg/mL에서 유의성 있는 세포 수 감소가 확인되었으므로 in vitro 상황에서 상기 추출물들의 최고 처리 농도를 1000 μg/mL로 결정하고, in vivo에서 사용할 농도는 동물의 체중(kg) 당 in vitro 최고 농도의 50배인 50 mg/kg으로 결정하였으며 5% DMSO가 포함된 PBS에 녹여 경구투여에 사용할 시료를 준비하였다.Cytotoxicity analysis of the extracts of Example 1 and Comparative Examples 1 and 2 using human chondrocyte showed that the highest cell concentration was reduced at the highest treatment concentration of 1000 μg / mL, and the concentration to be used in vivo was determined to be 50 mg / kg, which is 50 times the highest in vitro concentration per kg body weight of the animal. The sample to be used for oral administration was prepared by dissolving it in PBS containing 5% DMSO.
12마리의 SD rat을 3마리씩 5 group(normal, vehicle, 혼합 추출물 50 mg/kg, 옻나무 추출물 50 mg/kg, 노각나무 추출물 50 mg/kg)으로 나누어 1~2주 동안 adaptation 시킨다. Adaptation 후 normal group 3마리를 제외한 12마리의 rat을 대상으로 한쪽 무릎 관절 부위에 지름 2 mm의 드릴로 구멍을 뚫어 관절 연골이 손상된 OD(osteochondral defect) 모델을 구축하고, 다음 날 부터 체중 측정 후 vehicle 또는 50 mg/kg의 상기 추출물들을 각각 2일 간격으로 5주 동안 경구 투여하였다. 5주 후 동물모델을 희생하여 골관절 손상 부위를 포함한 뼈 조직을 분리하고, 분리된 뼈 조직을 포르말린을 사용하여 2주 동안 고정하여, 고정된 뼈 조직의 칼슘을 제거하여 부드럽게 만들어 주기 위해서 ~14% EDTA(pH 7.5)를 사용하여 de-calcification을 진행하였다. 칼슘 제거는 뼈 조직이 완전히 부드러워질 때까지 3일 간격으로 용액을 갈아주며 4~8주 정도 진행하였으며, 칼슘이 제거된 뼈 조직은 조직 처리과정을 거친 후 paraffin embedding하여 블록으로 만들고, 준비된 paraffin 블록은 골관절 손상 부위를 중심으로 섹션하여 염색을 위한 슬라이드를 준비하였다.Twelve SD rats were divided into 5 groups (normal, vehicle, mixed extract 50 mg / kg, Rhus verniciflua extract 50 mg / kg, beech tree extract 50 mg / kg) After adaptation, 12 rats except 3 normal group were drilled with a drill with a diameter of 2 mm on one knee joint to construct an osteochondral defect model with damaged articular cartilage, Or 50 mg / kg of the above extracts were orally administered at intervals of 2 days for 5 weeks. After 5 weeks, the animal model was sacrificed to separate the bone tissue including the injured area of the osteoarthritis. The bone tissue was fixed with formalin for 2 weeks to remove the calcium of the fixed bone tissue to make it smooth. De-calcification was carried out using EDTA (pH 7.5). The calcium removal was carried out for 4 to 8 weeks at 3-day intervals until the bone tissue was completely softened. The calcium-depleted bone tissue was subjected to tissue treatment and paraffin embedding to make a block. The prepared paraffin block Was prepared by slicing a section around the injured area of the osteotomy.
상기 준비된 조직 슬라이드를 safranin/fast green staining 방법으로 염색하여 연골의 손상, 회복 정도를 비교하였다(orang to red: cartilage, mucin, mast cell granules / bluish green-background: cytoplasm / black: nuclei).The prepared tissue slices were stained with safranin / fast green staining method to compare cartilage damage and recovery (orang to red: cartilage, mucin, mast cell granules / bluish green-background: cytoplasm / black: nuclei).
도 2는 실시예 1, 비교예 1 및 2에 따라 제조된 추출물을 투여 시 손상된 연골의 재생에 미치는 영향을 조직 염색법으로 분석한 결과이다.FIG. 2 shows the results of analysis of the effect of the extract prepared according to Example 1 and Comparative Examples 1 and 2 on the regeneration of injured cartilage by tissue staining.
도 2에 도시된 바와 같이, normal 그룹은 연골의 손상을 가하지 않은 그룹으로 매끈한 연골의 상태를 확인할 수 있었으며, 드릴로 연골에 손상을 가한 뒤 추출물 대신에 vehicle을 투여한 그룹은 연골의 손상이 심각하게 남아있음을 확인하였다.As shown in FIG. 2, the normal group was able to confirm the smooth cartilage state as a group that did not cause damage to the cartilage, and the group in which the vehicle was administered in place of the extract after damaging the cartilage with the drill, .
또한, 연골에 손상을 가한 뒤 비교예 1의 노각 추출물 및 비교예 2의 옻 추출물을 투여한 그룹에서도 연골의 손상이 남아있음을 확인하였다.In addition, after damage to the cartilage, it was confirmed that cartilage damage remained even in the group treated with the crude extract of Comparative Example 1 and the raspberry extract of Comparative Example 2.
반면, 연골에 손상을 가한 뒤 실시예 1의 혼합 추출물을 투여한 그룹에서는 드릴로 구멍을 뚫어 손상된 연골 부위가 매끈하게 차올라 있음을 확인할 수 있어, 실시예 1의 혼합 추출물가 뛰어한 연골 재생 능력을 나타냄을 보여준다.
On the other hand, after damaging the cartilage, it was confirmed that the cartilage area damaged by drilling was drilled smoothly in the group to which the mixed extract of Example 1 was administered, so that the mixed extract of Example 1 exhibited excellent cartilage regenerating ability Lt; / RTI >
따라서, 본 발명에 따라 제조된 혼합 추출물은 연골세포에 손상을 미치는 효소인 MMP-1, 3, 13의 유전자 발현을 효과적으로 억제하며, 손상된 연골을 재생시킬 수 있으므로 관절염을 예방하고 개선시키는데 현저한 효과를 나타낸다.
Therefore, the mixed extract prepared according to the present invention effectively inhibits the gene expression of MMP-1, 3, 13, which is an enzyme that damages cartilage cells, and regenerates damaged cartilage, thereby remarkably preventing and improving arthritis .
하기에 본 발명의 분말을 함유하는 조성물의 제제예를 설명하나, 본 발명은 이를 한정하고자 함이 아닌 단지 구체적으로 설명하고자 함이다.Hereinafter, formulation examples of the composition containing the powder of the present invention will be described, but the present invention is not intended to be limited thereto but is specifically described.
제제예 1. 산제의 제조Preparation Example 1. Preparation of powder
실시예 1에서 얻은 혼합 추출물 분말 500 mg500 mg of the mixed extract powder obtained in Example 1
유당 100 mg
탈크 10 mgTalc 10 mg
상기의 성분들을 혼합하고 기밀포에 충진하여 산제를 제조한다.
The above components are mixed and filled in airtight bags to prepare powders.
제제예 2. 정제의 제조Formulation Example 2. Preparation of tablets
실시예 1에서 얻은 혼합 추출물 분말 300 mg300 mg of the mixed extract powder obtained in Example 1
옥수수전분 100 mg
유당 100 mg
스테아린산 마그네슘 2 mgMagnesium stearate 2 mg
상기의 성분들을 혼합한 후 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조한다.
After mixing the above components, tablets are prepared by tableting according to the usual preparation method of tablets.
제제예 3. 캅셀제의 제조Formulation Example 3. Preparation of capsules
실시예 1에서 얻은 혼합 추출물 분말 200 mg200 mg of the mixed extract powder obtained in Example 1
결정성 셀룰로오스 3 mg
락토오스 14.8 mgLactose 14.8 mg
마그네슘 스테아레이트 0.2 mgMagnesium stearate 0.2 mg
통상의 캡슐제 제조방법에 따라 상기의 성분을 혼합하고 젤라틴 캡슐에 충전하여 캡슐제를 제조한다.
The above components are mixed according to a conventional capsule preparation method and filled in gelatin capsules to prepare capsules.
제제예 4. 주사제의 제조Formulation Example 4. Preparation of injection
실시예 1에서 얻은 혼합 추출물 분말 600 mg600 mg of the mixed extract powder obtained in Example 1
만니톨 180 mg180 mg mannitol
주사용 멸균 증류수 2974 mgSterile sterilized water for injection 2974 mg
Na2HPO4,12H2O 26 mgNa 2 HPO 4, 12H 2 O 26 mg
통상의 주사제의 제조방법에 따라 1 앰플 당 상기의 성분 함량으로 제조한다.
It is prepared by the above-mentioned component content per ampoule according to the usual injection preparation method.
제제예 5. 액제의 제조Formulation Example 5. Preparation of a liquid preparation
실시예 1에서 얻은 혼합 추출물 분말 4 g4 g of the mixed extract powder obtained in Example 1
이성화당 10 g10 g per isomer
만니톨 5 g5 g mannitol
정제수 적량Purified water quantity
통상의 액제의 제조방법에 따라 정제수에 각각의 성분을 가하여 용해시키고 레몬향을 적량 가한 다음 상기의 성분을 혼합한 다음 정제수를 가하여 전체를 정제수를 가하여 전체 100g으로 조절한 후 갈색병에 충진하여 멸균시켜 액제를 제조한다.
Each component was added and dissolved in purified water according to the usual liquid preparation method, and the lemon flavor was added in an appropriate amount. Then, the above components were mixed, and purified water was added thereto. The whole was adjusted to 100 g with purified water, To prepare a liquid agent.
제제예 6. 과립제의 제조Preparation Example 6 Preparation of Granules
실시예 1에서 얻은 혼합 추출물 분말 1,000 mg1,000 mg of the mixed extract powder obtained in Example 1
비타민 혼합물 적량Vitamin mixture quantity
비타민 A 아세테이트 70 ㎍70 [mu] g of vitamin A acetate
비타민 E 1.0 mgVitamin E 1.0 mg
비타민 B1 0.13 mgVitamin B1 0.13 mg
비타민 B2 0.15 mg0.15 mg of vitamin B2
비타민 B6 0.5 mgVitamin B6 0.5 mg
비타민 B12 0.2 ㎍0.2 [mu] g vitamin B12
비타민 C 10 mgVitamin C 10 mg
비오틴 10 ㎍Biotin 10 μg
니코틴산아미드 1.7 mgNicotinic acid amide 1.7 mg
엽산 50 ㎍50 ㎍ of folic acid
판토텐산 칼슘 0.5 mgCalcium pantothenate 0.5 mg
무기질 혼합물 적량Mineral mixture quantity
황산제1철 1.75 mg1.75 mg of ferrous sulfate
산화아연 0.82 mg0.82 mg of zinc oxide
탄산마그네슘 25.3 mgMagnesium carbonate 25.3 mg
제1인산칼륨 15 mgPotassium monophosphate 15 mg
제2인산칼슘 55 mgSecondary calcium phosphate 55 mg
구연산칼륨 90 mgPotassium citrate 90 mg
탄산칼슘 100 mg
염화마그네슘 24.8 mgMagnesium chloride 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 과립제에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 과립제 제조방법에 따라 상기의 성분을 혼합한 다음, 과립을 제조하고, 통상의 방법에 따라 건강기능식품 조성물 제조에 사용할 수 있다.
The composition ratio of the above-mentioned vitamins and minerals is comparatively comparatively mixed with the granules according to the preferred embodiment. However, the blending ratio may be arbitrarily changed, and the above components are mixed according to the ordinary granule preparation method, Can be prepared and used in the manufacture of a health functional food composition according to a conventional method.
제제예 7. 기능성 음료의 제조Preparation Example 7. Preparation of functional beverage
실시예 1에서 얻은 혼합 추출물 분말 1,000 mg 1,000 mg of the mixed extract powder obtained in Example 1
구연산 1,000 mgCitric acid 1,000 mg
올리고당 100 g100 g of oligosaccharide
매실농축액 2 gPlum concentrate 2 g
타우린 1 gTaurine 1 g
정제수를 가하여 전체 900 mLPurified water was added to the flask to obtain a total of 900 mL
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1 시간 동안 85 ℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 L 용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 기능성 음료 조성물 제조에 사용한다. The above components were mixed according to a conventional health drink manufacturing method, and the mixture was stirred and heated at 85 DEG C for about 1 hour. The solution thus prepared was filtered and sterilized in a sterilized 2 L container, It is used in the production of the functional beverage composition of the invention.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the composition ratio is a mixture of the components suitable for the preferred beverage as a preferred embodiment, the blending ratio may be arbitrarily varied according to the regional and national preferences such as the demand level, the demanding country, and the intended use.
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| KR20200124154A (en) * | 2019-04-23 | 2020-11-02 | 나천수 | A composition for improving, preventing and treating arthritis comprising extract of Stewartia pseudocamellia Maxim. leave and Cudrania tricuspidata leaves |
| WO2020218733A3 (en) * | 2019-04-23 | 2020-12-17 | 나천수 | Composition for alleviating, preventing or treating arthritis, containing stewartia pseudocamellia leaf extract and maclura tricuspidata leaf extract as active ingredients |
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| KR100945960B1 (en) | 2008-01-11 | 2010-03-05 | 주식회사 인섹트 바이오텍 | A composition containing Arazyme for the prevention and treatment of arthritis |
| JP2015027998A (en) | 2013-06-24 | 2015-02-12 | 株式会社東洋新薬 | Moisturizer, skin barrier function activator, tight junction formation accelerator, trpv4 expression enhancer, intracellular calcium concentration increaser, intracellular calcium concentration increaser, lipid synthesis accelerator, blood flow improver and periocular darkness ameliorator |
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| KR100807169B1 (en) * | 2006-09-20 | 2008-02-27 | 경희대학교 산학협력단 | A composition comprising an extract of rhus verniciflua stokes for the prevention and treatment of arthritis |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR100945960B1 (en) | 2008-01-11 | 2010-03-05 | 주식회사 인섹트 바이오텍 | A composition containing Arazyme for the prevention and treatment of arthritis |
| JP2015027998A (en) | 2013-06-24 | 2015-02-12 | 株式会社東洋新薬 | Moisturizer, skin barrier function activator, tight junction formation accelerator, trpv4 expression enhancer, intracellular calcium concentration increaser, intracellular calcium concentration increaser, lipid synthesis accelerator, blood flow improver and periocular darkness ameliorator |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20200124154A (en) * | 2019-04-23 | 2020-11-02 | 나천수 | A composition for improving, preventing and treating arthritis comprising extract of Stewartia pseudocamellia Maxim. leave and Cudrania tricuspidata leaves |
| WO2020218733A3 (en) * | 2019-04-23 | 2020-12-17 | 나천수 | Composition for alleviating, preventing or treating arthritis, containing stewartia pseudocamellia leaf extract and maclura tricuspidata leaf extract as active ingredients |
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