KR20080112722A - Paeonia japonica extracts showing a good therapeutic effect on osteoarthritis and its pharmaceutical compositions containing the same - Google Patents
Paeonia japonica extracts showing a good therapeutic effect on osteoarthritis and its pharmaceutical compositions containing the same Download PDFInfo
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Abstract
Description
도1은 연골세포에 백작약 추출물을 처리시 세포의 증식정도를 나타내는 결과이다.Figure 1 is a result showing the degree of proliferation of the cell when treated with chokbaek extract.
본 발명은, 퇴행성 관절염에 대해 치료효과가 우수한 백작약 추출물 및 그를 포함하는 약제학적 조성물에 관한 것이다. 퇴행성 관절염은 전체 근골격계 질환의 60% 정도를 차지하는 가장 흔한 운동기계의 질환이며 평균 연령이 증가하면서 유병율이 증가되고 있는 중요한 질환이다. (Meng J et al., Microarray analysis of differential gene expression in temporomandibular joint condylar cartilage after experimentally induced osteoarthritis. Osteoarthritis Cartilage. 2005, 13(12):1115-25; Zhang H et al. Microarray analysis reveals the involvement of beta-2 microglobulin (B2M) in human osteoarthritis. Osteoarthritis Cartilage. 2002, 10(12):950-60.) The present invention relates to a currant extract having excellent therapeutic effect against degenerative arthritis and a pharmaceutical composition comprising the same. Degenerative arthritis is the most common disease of the motor system, accounting for about 60% of all musculoskeletal disorders, and is an important disease with increasing prevalence as the average age increases. (Meng J et al ., Microarray analysis of differential gene expression in temporomandibular joint condylar cartilage after experimentally induced osteoarthritis. Osteoarthritis Cartilage. 2005, 13 (12): 1115-25; Zhang H et al . Microarray analysis reveals the involvement of beta- 2 microglobulin (B2M) in human osteoarthritis.Osteoarthritis Cartilage. 2002, 10 (12): 950-60.)
퇴행성 관절염이 발병되면 여러 가지 염증 세포의 침윤과 관절 내의 만성 염증 반응이 일어나고 pannus의 형성으로 인하여 연골조직의 파괴와 관절 뼈조직의 부식 및 변형을 초래하게 된다 (Cai L et al., Pathways by which interleukin 17 induces articular cartilage breakdown in vitro and in vivo. Cytokine. 2001, 16(1): 10-21). OA는 주로 체중부하 관절의 관절연골과 연부조직에서 만성적인 퇴 행성 변화를 가져오며, 퇴행성 변화에 의해 손상이 가속되는 양성질환으로 인식되어 왔다. 그러나 최근 유전적 인자, 개인적 또는 사회적 환경이 미치는 영향에 대탄 인식, 새로운 연구 기술의 발달 등으로 OA의 병인성 인자에 대한 연구가 활발하게 이루어지고 있다. (Haq Iet al., J. Osteoarthritis. Postgrad Med J. 2004,79(933) 377-383.)The development of degenerative arthritis leads to infiltration of various inflammatory cells and chronic inflammatory reactions in the joints, and the formation of pannus leads to the destruction of cartilage tissues and the corrosion and deformation of joint bone tissues (Cai L et al . , Pathways by which interleukin 17 induces articular cartilage breakdown in vitro and in vivo.Cytokine. 2001, 16 (1): 10-21). OA causes chronic degenerative changes in the articular cartilage and soft tissues of weight-bearing joints, and has been recognized as a benign disease in which damage is accelerated by degenerative changes. Recently, however, research on OA's pathogenic factors has been actively conducted due to anti-tank recognition on the effects of genetic factors, personal or social environment, and the development of new research techniques. (Haq I et al ., J. Osteoarthritis.Postgrad Med J. 2004,79 (933) 377-383.)
본 발명은 한약재의 일종인 백작약(白芍藥. Paeonia japonica)의 열탕, 또는 유기 용매 추출의 방법으로 제조되며, 퇴행성 관절염의 치료에 효과적인 백작약 추출물을 유효성분으로 포함하는 약제학적 조성물에 관한 것이다.The present invention is a kind of herbal medicine (baek 백. Paeonia) japonica ), or a method of extracting organic solvents, and relates to a pharmaceutical composition comprising as an active ingredient an extract of Paxenaceae effective for the treatment of degenerative arthritis.
본 발명은 이러한 목적을 달성하기 위하여 백작약를 정제추출하여 얻은 백작약 추출물 및 약학적으로 허용되는 백작약 추출 유효성분의 약제학적 조성물을 제공한다.The present invention provides a pharmaceutical composition of Baekjak extract and pharmaceutically acceptable Baekjak extract active ingredient obtained by purifying Baekjak to achieve this object.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명에 있어 백작약 고순도 추출물의 제조방법은 다음과 같다. 먼저 약재를 흐르는 증류수에 잘 세척한 후, 동일 부피의 증류수를 채워 충분히 불리고 반응전열기를 사용하여 열탕 추출한다. 0.45μm와 0.1μm의 filter paper를 연속적으로 사용하여 추출 액을 여과하고 약전소금을 이용하여 여과액 내의 무기염류들을 침전시켜 제거하고 citric acid와 Na3PO4를 이용하여 pH 7.3으로 조절한 후, 고압 살균하여 제조하였다.In the present invention, the manufacturing method of the earl high purity extract is as follows. First wash well with distilled water flowing through the medicine, and then filled with the same volume of distilled water is sufficiently called and extracted with hot water using a reaction heater. Filter the extract using 0.45μm and 0.1μm filter paper continuously, precipitate and remove inorganic salts in the filtrate using weak salt, adjust to pH 7.3 using citric acid and Na3PO4, and then autoclave Prepared.
또 다른 측면에서, 본 발명은, 백작약를 증류수와 혼합하여 가열 및 증류하여 여과 및 중탕하여 얻어진 백작약 추출물 유효성분의 염을 포함하는 것을 특징으로 하는 퇴행성 관절염의 치료용 약제학적 조성물을 제공한다.In another aspect, the present invention provides a pharmaceutical composition for the treatment of degenerative arthritis, characterized in that it comprises a salt of Baekjak extract active ingredient obtained by mixing Baekjak with distilled water, heating and distillation, filtered and bathed.
백작약 추출물의 유효성분을 고농도 농축하여 한의학적 치료에 사용되는 약침에 의해 투여되는 약침액으로 사용될 수 있다. 본 발명은, 상기 유효성분의 백작약 추출물을 관절 내의 염증 유발시 관절 내로의 세포 침윤을 억제하는 약제학적 조성물로 제공한다. 이러한 치료 방법은 내복약이 갖는 소화흡수 과정에서의 장시간 소요, 유효성분의 소실, 복용 후 위장장애 등의 부작용, 복용의 어려움 등을 극복하고 극소량의 고순도 약물로 경락경혈을 자극함으로서 약침의 기운이 내장의 병소에 직접 전달되어 최대의 효과를 볼 수 있으며 내성, 중독성, 습관성 등의 부작용을 줄일 수있는 장점이 있다. It can be used as a medicinal acupuncture solution administered by medicinal acupuncture used in Oriental medicine treatment by concentrating the active ingredient of Baekjak extract in high concentration. The present invention provides the bacterium extract of the active ingredient as a pharmaceutical composition that inhibits cell invasion into the joint when inflammation in the joint is induced. This treatment method overcomes the long-term use in the digestive absorption process of the oral medicine, loss of the active ingredient, side effects such as gastrointestinal disorders after taking the drug, difficulty in taking the medicine, and stimulates the acupuncture points with very high-purity drugs. It can be delivered directly to the lesions to see the maximum effect, and has the advantage of reducing side effects, such as resistance, addictive, and habitual.
본 발명에 사용된 백작약(白芍藥. Paeonia japonica)은 뿌리는 굵고 육질이며 밑부 분이 비늘 같은 잎으로 싸여 있다. 잎은 3∼4개가 어긋나고 3개씩 2번 갈라진다. 작은잎은 긴 타원형이거나 달걀을 거꾸로 세워놓은 모양이고 가장자리가 밋밋하며 털이 없다. 꽃은 6월에 흰색으로 피고 지름 4∼5cm이며 원줄기 끝에 1개씩 달린다. 꽃받침조각은 달걀 모양이며 3개, 꽃잎은 달걀을 거꾸로 세워놓은 모양이고 5∼7개이다. 수술은 여러 개이며 3∼4개의 씨방이 있다. 열매는 골돌과로서 벌어지면 안쪽이 붉고 덜 자란 붉은 종자와 성숙한 검은 종자가 나타난다. 몸의 氣(기)와 血(혈)이 함께 상한경우에 쓰는데 일반인들은 모두 감기약쯤으로 알고 있지만 그렇지 않은 처방이다. 몸의 음과 양의 상태를 잘 조화시켜 건강을 이끌어가는 명 처방이기에 제약회사들이 지금도 제품을 많이도 생산한다. 쌍화탕에서는 백작약이 바로 君藥(군약)인 주인 자리를 차지하고 나머지 약들은 작약을 보좌하는 상태로 처방이 구성되어있다. 이뿐만이 아니고 수많은 처방들에 작약이 들어가 있음을 알 수 있고 약용의 범위가 아주 넓게 적용된다고 할 수 있다. Paekonia japonica (白 芍藥. Paeonia japonica) used in the present invention, the roots are thick, fleshy, the base is wrapped with leaves like scales. Leaves are alternately 3-4 in number and split 2 times in 3 pieces. Small leaves are long oval or upside down, with flat edges and no hairs. Flower is white in June, 4 ~ 5cm in diameter, hang one by one at the end of main stem. Calyxes are egg-shaped, 3 pieces, petals are upside down, 5-7 pieces. There are several surgeries with three to four ovaries. Fruits are oligopods that, when opened, show reddish, less-grown red seeds and mature black seeds.氣 (기) and 血 (blood) of the body is used together when the upper limit, the common people all know about cold medicine, but it is not a prescription. Pharmaceutical companies are still producing a lot of products because it is a prescription that leads health by harmonizing the yin and yang of the body well. In Ssanghwatang, the Count is the owner of the 君 藥 (military medicine) and the rest of the medicine is composed of a prescription to support the peony. Not only that, but many prescriptions contain peony, and the range of medicinal application is very wide.
Ⅰ. 실 험 방 법I. Experiment method
MTT assay: MTT assay에 사용할 돼지의 연골세포를 96well plate에 1×104cell/well이 되도록 분주한다. 세포가 잘 부착 할 수 있게 24시간동안 37℃ 5% CO2 incubator배양 한다. 그 후 처리할 약물을 농도에 맞춰 각 well에 분주 한 후 적정 시간 동안 37℃ 5% CO2 incubator에서 배양 한다. media에 MTT(Sigma, USA)를 넣어 1%용액으로 만든 후 각 well당 20ul씩 처리 하고 세시간 동안 37℃ 5% CO2 incubator에서 배양한다. 세 시간 후 상층액을 모두 버린 후 acid isopropanol (HCl 660ul, isopropanol 200ml)을 well당 100ul씩 처리하였다. 37℃ shaking incubator에서 5분간 shaking 후 570nm에서 O.D값을 측정하였다. MTT assay: Dispense pig chondrocytes to be 1 × 10 4 cells / well in a 96well plate for MTT assay. Incubate at 37 ° C for 5% CO2 incubator for 24 hours to allow cells to adhere well. Subsequently, the drug to be treated is dispensed into each well according to the concentration, and then incubated in a 37% 5% CO2 incubator for a predetermined time. Add MTT (Sigma, USA) to the media, make 1% solution, process 20ul for each well, and incubate in 37 ℃ 5% CO2 incubator for 3 hours. After three hours, the supernatant was discarded and treated with 100 isul per acid isopropanol (HCl 660ul, isopropanol 200ml). After shaking for 5 minutes at 37 ℃ shaking incubator and measured the OD value at 570nm.
II. 결론II. conclusion
MTT assay의 결과를 보면(도 1) 백작약가 연골세포의 증식을 대조군에 비해 186% 의 증식효과를 보여 주었다. The results of the MTT assay (Fig. 1) showed that the ear of baekchaek showed a proliferative effect of 186% compared to the control of the chondrocytes.
이상의 결과와 같이, 백작약 추출액 처치는 퇴행성 관절염에 의한 연골세포 손상을 억제시키고, 연골세포의 성장을 촉진하는 효능이 있다. 따라서 대표적인 퇴행성 관절염의 새로운 치료 보조제임을 알 수 있다. As described above, the treatment of Baekjak extract has the effect of inhibiting chondrocyte damage caused by degenerative arthritis and promoting the growth of chondrocytes. Therefore, it can be seen that it is a new treatment aid of representative degenerative arthritis.
이상 실험 예를 들어 본 발명을 상세히 설명하였으나, 본 발명은 이에 제한되지 않는다. Although the present invention has been described in detail with reference to the above experimental examples, the present invention is not limited thereto.
위 실시 예에서 제재는 백작약 추출물의 고순도 추출액인 것으로 설명하였으나 통상적으로 약제학적으로 허용되는 부형제와 함께 약제학적으로 허용되는 약제학 제재, 예를 들어 주사제, 액제, 시럽제, 정제, 캡슐제 등으로 체계화하여 퇴행성 관 절염과 같은 만성자가면역질환의 예방 및 치료를 위한 약학적 제재로 사용될 수 있다. In the above embodiment, the preparation is described as a high purity extract of the earl extract, but is usually formulated with a pharmaceutically acceptable excipient together with a pharmaceutically acceptable pharmaceutical preparation, for example, injections, solutions, syrups, tablets, capsules, etc. It can be used as a pharmaceutical agent for the prevention and treatment of chronic autoimmune diseases such as degenerative arthritis.
본 발명에 따른 제재는 임상적 투여시 경구, 복강, 피하 투여 등의 방법으로 투여될 수도 있다.The agent according to the present invention may be administered by a method such as oral, intraperitoneal, subcutaneous administration during clinical administration.
본 발명에 따른 제재의 임상적 투여량은 환자의 연령, 증상, 투여제형, 또는 약물의 종류에 따라 다양하게 조절할 수 있다. 특정한 상태에서 바람직한 투여량을 결정하는 것은 공지된 기술에 해당한다.The clinical dosage of the preparations according to the invention can be varied according to the age, symptoms, dosage form, or type of drug of the patient. Determining the desired dosage in a particular condition corresponds to known techniques.
본 발명은, 퇴행성 관절염의 주원인 세포인 연골세포에 백작약 고순도 증류 추출액을 투여한 결과, 세포의 증식을 활성화시킴을 알 수 있다. 즉 백작약 추출액 처치가 퇴행성 관절염의 치료에 효과적임을 알 수 있다.According to the present invention, the high purity distillate extract was administered to chondrocytes, which are the main cells of degenerative arthritis, thereby activating proliferation of the cells. That is, it can be seen that Baekja extract treatment is effective in the treatment of degenerative arthritis.
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| JP2013503155A (en) * | 2009-08-31 | 2013-01-31 | 株式会社アモーレパシフィック | Method for promoting skin regeneration effect of chemicals using pottery bath |
| CN111093398A (en) * | 2017-09-14 | 2020-05-01 | 韩国韩医学研究院 | Composition for preventing, improving or treating osteoarthritis containing Alpinia oxyphylla extract as active ingredient |
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| JP2013503155A (en) * | 2009-08-31 | 2013-01-31 | 株式会社アモーレパシフィック | Method for promoting skin regeneration effect of chemicals using pottery bath |
| CN111093398A (en) * | 2017-09-14 | 2020-05-01 | 韩国韩医学研究院 | Composition for preventing, improving or treating osteoarthritis containing Alpinia oxyphylla extract as active ingredient |
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