KR100997215B1 - The pharmaceutical composition comprising the extract of Astragalus membrancens Bunge or ethyl acetate fraction therefrom for treat and prevantion for bone growth trouble - Google Patents
The pharmaceutical composition comprising the extract of Astragalus membrancens Bunge or ethyl acetate fraction therefrom for treat and prevantion for bone growth trouble Download PDFInfo
- Publication number
- KR100997215B1 KR100997215B1 KR1020080009540A KR20080009540A KR100997215B1 KR 100997215 B1 KR100997215 B1 KR 100997215B1 KR 1020080009540 A KR1020080009540 A KR 1020080009540A KR 20080009540 A KR20080009540 A KR 20080009540A KR 100997215 B1 KR100997215 B1 KR 100997215B1
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- KR
- South Korea
- Prior art keywords
- astragalus
- extract
- growth
- ethyl acetate
- acetate fraction
- Prior art date
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- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
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Abstract
본 발명은 황기(Astragalus membrancens Bunge) 추출물 또는 에틸아세테이트 분획물을 포함하는 골길이 성장 장애 치료 및 예방용 약학조성물에 관한 것으로, 황기 추출물 및 에틸아세테이트 분획물은 효과적으로 골길이 성장을 촉진하므로 소인증, 왜소증, 키 성장 저하 등 골길이 성장 장애의 치료 및/또는 예방을 위한 골길이 성장 장애 치료 및 예방용 약학조성물 및 건강식품으로 유용하게 이용될 수 있다.The present invention is Astragalus membrancens Bunge ) The present invention relates to a pharmaceutical composition for treating and preventing bone length growth disorders comprising an extract or an ethyl acetate fraction. It can be usefully used as a pharmaceutical composition and health food for treating and preventing bone length growth disorders for the treatment and / or prevention of disorders.
생약추출물, 성장, 황기 Herbal extracts, growth, Astragalus
Description
본 발명은 황기 추출물 또는 이로부터 분리된 에틸아세테이트 분획물을 포함하는 골길이 성장 장애 치료 및 예방용 약학조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for treating and preventing bone length growth disorders comprising an extract of Astragalus or an ethyl acetate fraction isolated therefrom.
성장이란 신장의 증가를 말하며, 영양과 성장호르몬 등에 의하여 촉진된다. 특히 성장호르몬을 분비하는 뇌를 포함한 신경계는 유년기에 현저하게 생장하고, 그 후는 성장이 완만해진다. 따라서, 사람의 성장은 대부분 사춘기까지의 시기에 대부분 일어나며, 이 시기에 적절한 영양섭취가 이루어지지 않으면 성장이 제대로 이루어지지 않게 된다. 이에 영양의 균형화를 이루기 위하여 많은 성장 촉진용 건강식품이 출시되고 있으나, 단순히 성장을 위하여 도움이 되는 여러 가지 성분들의 조합에만 그침으로써 아직도 그 효과가 만족할 만한 수준에 도달하지 못하였다.Growth refers to the increase in height, and is promoted by nutrition and growth hormone. In particular, the nervous system, including the brain that secretes growth hormone, grows remarkably in childhood, and then grows slowly. Therefore, most of human growth occurs until the age of puberty, and if proper nutrition is not achieved at this time, the growth will not be done properly. In order to achieve nutritional balance, many growth-promoting health foods have been released, but only the combination of various ingredients helpful for growth has not yet reached a satisfactory level.
장골의 길이 성장은 신체의 신장과 골격을 결정지으며 특별한 기작에 의해 조절되어진다. 특히 장골의 근위부성장판(epiphyseal growth plate)의 성장이 골의 길이 성장과정에 가장 중요한 척도가 된다. 성장판은 연골세포(chondrocyte)를 세포증식이 일어나기 전인 휴지부(resting zone)와 증식시키는 증식부(proliferation zone)와 연골세포의 성숙과 비대작용을 하는 성숙부(maturation zone)와 비대부(hypertrophic zone)로 나누어지고, 이들의 상호작용으로 장골의 길이성장이 이루어진다(Loveridge, N, J. Anim. Sci. 77, 190-196, 1999).The growth of the long bone determines the body's height and skeleton and is controlled by special mechanisms. In particular, the growth of the epiphyseal growth plate of the iliac bone is the most important measure in the bone growth process. The growth plate is a resting zone that proliferates chondrocytes before proliferation, a proliferation zone that proliferates, and a maturation zone and hypertrophic zone that matures and enlarges chondrocytes. And their interactions lead to long bone growth (Loveridge, N, J. Anim. Sci. 77, 190-196, 1999).
성장에 필요한 성장호르몬은 오래 전부터 왜소증(矮小症) 환자들의 치료목적으로 사용되어 왔고, 1985년 후반기에 이르러 유전자 재조합 방식의 성장호르몬이 상품화되기 시작하여 대량 생산 및 공급이 가능해지고 타질환의 위험성도 없어짐에 따라 최근에는 유전자 재조합 방식의 성장호르몬이 성장호르몬 결핍 질환의 치료제로 사용되고 있으며, 생체내의 엔케팔린(enkephalin)을 토대로 성장호르몬 분비활성을 갖는 6개의 아미노산으로 구성된 새로운 성장호르몬 분비 촉진제인 GHRP-6(Growth hormone releasing peptide-6, His-D-Trp-Ala-Trp-D-Phe-Lys-NH2)가 개발되어(Bowers CY. et al., Endocrinology, 114, 1537-45, 1984), 비록 적은 흡수율이지만 경구 투여시에도 흡수가 되는 것으로 보고되었다(Noriko I. et al ., Life Science, 47, 29-36, 1990). 이후에 계속적인 연구로 다른 펩티드성 성장호르몬 분비 촉진제인 GHRP-1(Ala-His-D-β-Nal-Ala-Trp-D-Phe-Lys-NH2)과 GHRP-2(D-Ala-D-β-Nal- Ala-Trp-D-Phe-Lys-NH2)라는 6개의 변화된 펩티드로 구성된 성장호르몬 분비 촉진제가 개발되어 보고되었다. 그러나 이들은 생체 내의 활성은 뛰어나지만 경구 투여시 그 흡수율이 2 내지 3 %에 머무르는 등 경구 투여에는 개선의 여지가 많은 것으로 보고되었다.Growth hormone required for growth has been used for the treatment of dwarfism patients for a long time.In late 1985, the recombinant growth hormone was commercialized, and mass production and supply became possible, and the risk of other diseases Recently, genetically modified growth hormone has been used as a treatment for growth hormone deficiency disease, and GHRP-6, a new growth hormone secretagogue consisting of 6 amino acids with growth hormone secretion activity based on enkephalin in vivo (Growth hormone releasing peptide-6, His-D-Trp-Ala-Trp-D-Phe-Lys-NH2) has been developed (Bowers CY. Et al., Endocrinology , 114, 1537-45, 1984), although Absorption rate, but has been reported to be absorbed upon oral administration (Noriko I. et. al ., Life Science , 47, 29-36, 1990). Subsequent studies continued with other peptide growth hormone secretion promoters, GHRP-1 (Ala-His-D-β-Nal-Ala-Trp-D-Phe-Lys-NH2) and GHRP-2 (D-Ala-D A growth hormone secretagogue consisting of six modified peptides, -β-Nal-Ala-Trp-D-Phe-Lys-NH2), has been developed and reported. However, they have been reported to have a lot of room for improvement in oral administration, such as excellent in vivo activity, but the absorption rate stays at 2-3% upon oral administration.
한편, 황기는 콩과(豆科, Leguminosae)에 속한 다년생 초본인 황기(astragalus membranceus Bunge)의 주피를 거의 벗긴 뿌리로, 성분으로는 자당, 글루코론산(glucoronic acid), 다종의 아미노산, 고미질, 점액질, 콜린(choline), 베타인(betaine) 및 엽산 등을 포함한다. 전통 한의학에서의 효능으로는 보기요약으로서 신체허약, 자한(自汗), 종기, 지갈(止渴), 복통, 신쇠(腎衰), 이농(耳聾), 허천(虛喘), 배농지통(排膿止痛), 음식무미(飮食無味), 식욕부진, 치질, 목적(目赤), 학질(栖疾), 탈항(脫肛), 자궁출혈, 부인대하, 월경불순, 산전산후일절병, 거담수(祛痰嗽), 두통, 석폐심화(潟肺心火), 이질 및 소아백병 등에 유효하다. 문헌으로는 조혈작용(Ma R. et al ., J Tradit Chin Med. Sep; 3(3):199-204, 1983), 항산화작용(이춘영, 강원대 농업과학연구소 논문집 15:103, 2004), 토끼의 조골세포분화에 미치는 효과(Xu CJ. et al., Zhong Nan Da Xue xue Bao Yi Xue Ban, Aug;29(4):489-91, 2004), 난소적출쥐에서의 골다공증예방효과(Kim C. et al ., Arch Pharm Res. Nov;26(11):917-924, 2003) 등이 있으나, 성장판의 길이성장에 대하여 연구된 바는 없는 실정이다.On the other hand, Astragalus is a root almost stripped of the pericarp of the perennial herbaceous Astragalus membranceus Bunge, which belongs to Leguminosae. Mucus, choline, betaine and folic acid. The benefits of traditional Korean medicine include boils, body weakness, jahan, boil, jigal, abdominal pain, kidney pain, diurnal disease, vulgaris and drainage pain.膿 止痛), food taste, loss of appetite, hemorrhoids, purpose, malaria, disinfection, uterine bleeding, gynecological disorders, menstrual irregularities, postpartum ectopic disease, expectorant water (祛痰 嗽), headache, septic deepening (潟肺 心火), dysentery and pediatric leukemia is effective. The literature describes hematopoiesis (Ma R. et. al ., J Tradit Chin Med . Sep; 3 (3): 199-204, 1983), antioxidant activity (Chun-Young Lee, Kangwon National University Research Institute of Agricultural Science 15: 103, 2004), effect on osteoblast differentiation in rabbits (Xu CJ. Et al., Zhong Nan Da Xue xue Bao Yi Xue Ban, Aug; 29 (4): 489-91, 2004), Preventing Osteoporosis in Ovarian Rats (Kim C. et. al ., Arch Pharm Res . Nov; 26 (11): 917-924, 2003), but there is no study on the growth of the length of the growth plate.
본 발명자는 성장 촉진에 이용되는 성장호르몬의 고비용, 저흡수율 및 부작용 등의 문제점을 해결하고자, 천연물을 대상으로 검색하던 중 황기 추출물 또는 에틸아세테이트 분획물이 피하 주사하는 성장호르몬보다 성장에 우수한 효과가 있음을 확인하여 본 발명을 완성하였다.In order to solve the problems such as high cost, low absorption rate and side effects of growth hormone used for growth promotion, the present inventors have an excellent effect on growth than growth hormone injected by subcutaneous extract or ethyl acetate fraction while searching for natural products. It was confirmed to complete the present invention.
상기 목적을 달성하기 위하여, 본 발명은 황기 추출물 및 이로부터 분리된 에틸아세테이트 분획물 및 약학적으로 허용되는 담체로 구성되는 골길이 성장 장애 치료 및 예방용 약학조성물을 제공한다.In order to achieve the above object, the present invention provides a pharmaceutical composition for the treatment and prevention of bone length growth disorders consisting of the extract of Astragalus and the ethyl acetate fraction separated therefrom and a pharmaceutically acceptable carrier.
또한 본 발명은 상기 약학조성물을 유효성분으로 함유하는 골길이 성장 촉진용 건강식품을 제공한다.In another aspect, the present invention provides a healthy food for promoting bone length growth containing the pharmaceutical composition as an active ingredient.
본 발명의 황기 추출물 또는 에틸아세테이트 분획물은 성장에 효과를 나타내므로, 소인증, 왜소증, 키 성장 저하 등 골길이 성장 장애의 치료 및 예방을 위한 촉진용 조성물에 이용될 수 있을 뿐 아니라 건강식품으로도 활용 가능하다. Astragalus extract or ethyl acetate fraction of the present invention exhibits an effect on growth, can be used in the composition for the promotion and treatment of bone length growth disorders, such as small authentication, dwarfism, height growth, as well as health food It can be utilized.
본 발명은 황기 추출물 및 이로부터 분리된 에틸아세테이트 분획물 및 약학적으로 허용되는 담체로 구성되는 골길이 성장 장애 치료 및 예방용 약학조성물을 제공한다.The present invention provides a pharmaceutical composition for the treatment and prevention of bone length growth disorders consisting of the Astragalus extract and ethyl acetate fractions isolated therefrom and a pharmaceutically acceptable carrier.
생약 추출물은 황기(경동시장, 서울)를 건조하여 마쇄한 후, 건조된 시료의 중량의 약 1 내지 20배, 바람직하게는 약 5 내지 15 배, 더욱 바람직하게는 10배 분량의 물, 메탄올, 에탄올 등과 같은 C1 내지 C4의 저급 알코올 또는 물과 저급 알코올의 혼합용매인 것이 바람직하며, 상기 혼합용매의 농도는 10 ~ 90% 알코올인 것이 바람직하며, 70% 알코올인 것이 더욱 바람직하다. 실온에서 약 1 내지 24시간, 바람직하게는 5 내지 15시간 동안, 가장 바람직하게는 6시간 동안 추출하는 것이다. 추출 방법으로는 열수 추출, 냉침 추출, 환류 냉각 추출 또는 초음파 추출 등이 바람직하며, 더욱 바람직하게는 환류냉각 추출한 후 감압 농축함으로써 본 발명의 가용 추출물인 조추출물을 수득할 수 있다. The herbal extracts are dried and crushed by the yellow rice (Gyeongdong Market, Seoul), and then about 1 to 20 times, preferably about 5 to 15 times, more preferably 10 times the amount of water, methanol, It is preferable that the lower alcohol of C 1 to C 4 such as ethanol or a mixed solvent of water and lower alcohol, the concentration of the mixed solvent is preferably 10 to 90% alcohol, more preferably 70% alcohol. Extraction at room temperature for about 1 to 24 hours, preferably 5 to 15 hours, most preferably 6 hours. As the extraction method, hot water extraction, cold needle extraction, reflux cooling extraction or ultrasonic extraction is preferable, and more preferably, the crude extract which is the soluble extract of the present invention can be obtained by reflux-cooling extraction and concentration under reduced pressure.
조추출물 100 g을 1 L의 70% 에탄올에 현탁시킨 후 각각 1 L의 헥산, 에틸아세테이트, 물포화부탄올로 단계적으로 용매의 극성을 높여가면서 각 용매의 가용분획별로 3회씩 분획한 다음, 에틸아세테이트 분획층을 모아 회전 감압 농축기로 완전 농축하고 동결건조하였다. 그 결과 3.1 g의 에틸아세테이트 분획 분말을 얻을 수 있었다. 100 g of crude extract was suspended in 1 L of 70% ethanol, and then fractionated three times for each soluble fraction of each solvent, increasing the polarity of the solvent stepwise with 1 L of hexane, ethyl acetate, and water saturated butanol, and then ethyl acetate. Fractions were combined, concentrated completely with a rotary vacuum concentrator, and lyophilized. As a result, 3.1 g of ethyl acetate fraction powder was obtained.
황기 추출물 또는 에틸아세테이트 분획물의 성장 측정을 위하여, 흰쥐를 음성 대조군, 황기 추출물(실험군 1 내지 6군), 황기 에틸아세테이트 분획물(실험군 7군) 및 양성 대조군으로 나누어 매일 2회씩 4일 동안 음성 대조군에게는 식염수를, 실험군에게는 황기 추출물을 각각 100 ㎎/kg 및 500 ㎎/kg 농도로 경구 투여하였고, 양성 대조군에게는 성장호르몬을 피하 주사하였다. 성장은 성장판에 테트라사이클린(tetracycline)이 침착된 후 성장한 경골부의 길이로 알 수 있으며, 이는 테트라사이클린의 침착으로 발색된 발광선과 성장판과의 간격을 측정함으로써 알 수 있다. 이를 위하여, 3일째에는 흰쥐에 테트라사이클린을 복강 주사하였고 그 후, 48 시간 경과한 5일째에는 흰쥐들을 에테르로 마취하여 희생하였다. 이후, 흰쥐에서 테트라사이클린으로 침착되어 발색된 발광선과 성장판과의 간격을 형광현미경으로 촬영한 후, 골길이 성장을 측정하였다(도 1 참조). 그 결과, 황기 추출물 또는 에틸아세테이트 분획물은 골길이 성장에 유의한 영향을 주는 것으로 나타났다. 황기 추출물 투여군(실험군 1 내지 실험군 6) 및 황기 에틸아세테이트 분획물의 골길이 성장은 음성 대조군의 골길이 성장에 비하여 통계적으로 높았으며, 양성 대조군보다도 우수한 성장 효능을 보였다. **는 P< 0.01을 나타낸다. 또한 식염수를 투여한 음성 대조군의 성장을 100%로 가정할 경우, 황기 추출물 및 황기 에틸아세테이트 분획물을 투여한 실험군의 골길이 성장률은 성장호르몬을 투여한 양성 대조군의 골길이 성장률보다 높게 나타났다(표 1 및 도 3 참조). To measure the growth of Astragalus extract or ethyl acetate fraction, rats were divided into a negative control, Astragalus extract (Experimental group 1 to 6), Astragalus ethyl acetate fraction (Experimental group 7) and a positive control twice daily for 4 days. Saline was administered orally to the experimental group at concentrations of 100 mg / kg and 500 mg / kg, respectively, and the positive control group was injected subcutaneously with growth hormone. Growth can be seen by the length of the tibial portion grown after the tetracycline (tetracycline) is deposited on the growth plate, it can be seen by measuring the distance between the growth plate and the light emitting color developed by the deposition of tetracycline. To this end, rats were intraperitoneally injected with tetracycline on day 3 and then rats were sacrificed with ether anesthesia on day 5 after 48 hours. Subsequently, after photographing the interval between the growth plate deposited with tetracycline in the rat and the growth plate by fluorescence microscope, bone length growth was measured (see FIG. 1). As a result, Astragalus extract or ethyl acetate fraction was found to have a significant effect on bone length growth. The bone length growth of the Astragalus extract administration group (Experimental Group 1 to Experimental Group 6) and Astragalus ethyl acetate fractions was statistically higher than that of the negative control group, showing better growth efficiency than the positive control group. ** indicates P <0.01. Also, assuming 100% growth of the negative control group administered with saline, the bone length growth rate of the experimental group administered with Astragalus extract and Astragalus ethyl acetate fraction was higher than that of the positive control group treated with growth hormone (Table 1). And FIG. 3).
상기의 황기 추출물 또는 에틸아세테이트 분획물을 마우스에 경구 투여하여 독성 실험을 수행한 결과, 경구 투여 독성시험에 의한 50% 치사량(LD50)은 적어도 1,000 ㎎/㎏ 이상인 안전한 물질로 판단된다.As a result of oral administration of the above extract or the ethyl acetate fraction to mice, 50% lethal dose (LD 50 ) by oral toxicity test was determined to be a safe substance of at least 1,000 mg / kg or more.
상기 결과를 통하여 황기 추출물 또는 에틸아세테이트 분획물은 안전한 물질이며, 성장호르몬보다 우수한 골길이 성장의 효과를 나타내므로 골길이 성장 장애 치료 및 예방용 약학조성물로서 제공될 수 있다. Through the above results, Astragalus extract or ethyl acetate fraction is a safe substance, and because it shows an effect of bone length growth superior to growth hormone, it can be provided as a pharmaceutical composition for treating and preventing bone length growth disorders.
본 발명의 골길이 성장 장애 치료 및 예방용 약학조성물은 상기 황기 추출물 및 황기 에틸아세테이트 분획물을 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.The pharmaceutical composition for treating and preventing bone length growth disorders of the present invention may further include appropriate carriers, excipients, and diluents which are commonly used in the preparation of the composition for extracting the Astragalus extract and Astragalus ethylacetate.
본 발명의 황기 추출물 및 황기 에틸아세테이트 분획물의 약학적 투여 형태는 이들의 약학적 허용 가능한 염의 형태로도 사용될 수 있고, 또한 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 집합으로 사용될 수 있다. Pharmaceutical dosage forms of the Astragalus extract and Astragalus ethylacetate fraction of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection.
본 발명의 조성물은 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 외용제, 좌제 및 멸균 주사용액의 형태로 제형화하여 사용될 수 있다. 목초액을 함유하는 조성물에 포함될 수 있는 담체, 부형제 및 희석제로는 락토즈, 덱스트로즈, 수크로스, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로스, 폴리비닐 피롤리돈, 물, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 제제화할 경우에는 보통 사용하는 충진제, 증량제, 결합제, 습윤제, 붕해제, 계면활성제 등의 희석제 또는 부형제를 사용하여 조제된다.The compositions of the present invention can be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols and the like, oral formulations, external preparations, suppositories, and sterile injectable solutions. Carriers, excipients, and diluents that may be included in compositions containing wood vinegar include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate , Cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.
경구투여를 위한 고형제제에는 정제, 환제, 산제, 과립제, 캡슐제 등이 포함 되며, 이러한 고형제제는 상기 생약 추출액에 적어도 하나 이상의 부형제, 예를 들면, 전분, 칼슘카보네이트(calcium carbonate), 수크로오스(sucrose) 또는 락토오스(lactose), 젤라틴 등을 섞어 조제된다. 또한 단순한 부형제 이외에 마그네슘 스테아레이트, 탈크 같은 윤활제들도 사용된다. 경구를 위한 액상 제제로는 현탁제, 내용액제, 유제, 시럽제 등이 해당되는데 흔히 사용되는 단순희석제인 물, 리퀴드 파라핀 이외에 여러 가지 부형제, 예를 들면 습윤제, 감미제, 방향제, 보존제 등이 포함될 수 있다.Solid preparations for oral administration include tablets, pills, powders, granules, capsules, and the like, and the solid preparations may include at least one excipient such as starch, calcium carbonate, sucrose ( It is prepared by mixing sucrose or lactose and gelatin. In addition to simple excipients, lubricants such as magnesium stearate and talc are also used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups, and may include various excipients, such as wetting agents, sweeteners, fragrances, and preservatives, in addition to commonly used simple diluents such as water and liquid paraffin. .
비경구 투여를 위한 제제에는 멸균된 수용액, 액제, 비수성용제, 현탁제, 유제, 동결건조 제제, 좌제, 주사제제가 포함된다. 비수성용제, 현탁제로는 프로필렌글리콜(propylene glycol), 폴리에틸렌 글리콜, 올리브 오일과 같은 식물성 기름, 에틸올레이트와 같은 주사 가능한 에스테르 등이 사용될 수 있다. 좌제의 기제로는 위텝솔(witepsol), 마크로골, 트윈(tween) 61, 카카오지, 라우린지, 글리세로제라틴 등이 사용될 수 있다.Formulations for parenteral administration include sterile aqueous solutions, solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, suppositories, injections. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate and the like can be used. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.
본 발명의 황기 추출물 및 에틸아세테이트의 바람직한 투여량은 환자의 상태 및 체중, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나 바람직한 효과를 위해서, 본 발명의 황기 추출물 또는 에틸아세테이트 분획물은 1일 0.0001 내지 500 ㎎/㎏으로, 바람직하게는 0.001 내지 500 ㎎/㎏으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The preferred dosage of the Astragalus extract and ethyl acetate of the present invention depends on the condition and weight of the patient, the extent of the disease, the form of the drug, the route of administration and the duration, and can be appropriately selected by those skilled in the art. However, for the desired effect, the Astragalus extract or ethyl acetate fraction of the present invention is preferably administered at 0.0001 to 500 mg / kg, preferably at 0.001 to 500 mg / kg. Administration may be administered once a day or may be divided several times. The dosage does not limit the scope of the invention in any aspect.
본 발명의 황기 추출물 또는 에틸아세테이트 분획물은 쥐, 생쥐, 가축, 인간 등의 포유동물에 다양한 경로로 투여될 수 있다. 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내(intracerebroventricular) 주사에 의해 투여될 수 있다. Astragalus extract or ethyl acetate fraction of the present invention can be administered to various mammals such as rats, mice, livestock, humans. All modes of administration may be expected, for example, by oral, rectal or intravenous, intramuscular, subcutaneous, intra-uterine or intracerebroventricular injections.
본 발명의 황기 추출물 또는 에틸아세테이트 분획물은 독성 및 부작용은 거의 없으므로 장기간 복용 시에도 안심하고 사용할 수 있는 조성물이다.Astragalus extract or ethyl acetate fraction of the present invention is a composition that can be used safely even when taking long-term because there is little toxicity and side effects.
또한 본 발명은 상기 조성물을 유효성분으로 함유하는 골길이 성장 촉진용 건강식품을 제공한다.In another aspect, the present invention provides a health food for promoting bone length growth containing the composition as an active ingredient.
황기 추출물 또는 에틸아세테이트 분획물을 첨가할 수 있는 식품으로는, 예를 들어, 각종 식품류, 유제품, 음료, 껌, 차, 비타민 단일제, 건강보조 식품류 등이 있고, 분말, 과립, 정제, 캡슐 또는 음료인 형태로 사용할 수 있다.Foods to which the Astragalus extract or ethyl acetate fraction can be added include, for example, various foods, dairy products, beverages, gums, teas, vitamin monosaccharides, dietary supplements, and the like, which are powders, granules, tablets, capsules or beverages. Available in form.
또한, 성장 촉진의 효과를 목적으로 식품 또는 음료에 첨가될 수 있다. 이 때, 식품 또는 음료 중의 상기 추출물의 양은 전체 식품 중량의 0.01 내지 15 중량%로 가할 수 있으며, 건강 음료 조성물은 100 ㎖를 기준으로 0.02 내지 5 g, 바람직하게는 0.3 내지 1 g의 비율로 가할 수 있다. It may also be added to foods or beverages for the purpose of promoting growth. At this time, the amount of the extract in the food or beverage can be added in 0.01 to 15% by weight of the total food weight, the health beverage composition is added in a ratio of 0.02 to 5 g, preferably 0.3 to 1 g based on 100 ml Can be.
본 발명의 건강 기능성 음료 조성물은 지시된 비율로 필수 성분으로서 상기 황기 추출물 또는 에틸아세테이트 분획물을 함유하는 외에는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물의 예는 모노사카라이드, 예 를 들어, 포도당, 과당 등; 디사카라이드, 예를 들어 말토스, 수크로오스 등; 및 폴리사카라이드, 예를 들어 덱스트린, 시클로덱스트린 등과 같은 통상적인 당, 및 자일리톨, 소르비톨, 에리트리톨 등의 당알코올이다. 상술한 것 이외의 향미제로서 천연 향미제(타우마틴, 스테비아 추출물(예를 들어 레바우디오시드 A, 글리시르히진등) 및 합성 향미제(사카린, 아스파르탐 등)를 유리하게 사용할 수 있다. 상기 천연 탄수화물의 비율은 본 발명의 조성물 100 ㎖당 일반적으로 약 1 내지 20 g, 바람직하게는 약 5 내지 12 g이다.The health functional beverage composition of the present invention is not particularly limited to other ingredients other than the above containing Astragalus extract or ethyl acetate fraction as essential ingredients in the indicated ratios, and various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks, may be used as additional ingredients. It may contain. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; Disaccharides such as maltose, sucrose and the like; And conventional sugars such as polysaccharides such as dextrin, cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those mentioned above, natural flavoring agents (tauumatin, stevia extract (for example, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of said natural carbohydrates is generally about 1-20 g, preferably about 5-12 g per 100 ml of the composition of the present invention.
상기 외에 본 발명의 황기 추출물 또는 에틸아세테이트 분획물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 중진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그밖에 본 발명의 황기 추출물 또는 에틸아세테이트 분획물은 천연 과일 주스 및 과일 주스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 추출물 또는 에틸아세테이트 분획물 100 중량부 당 0.1 내지 약 20 중량부의 범위에서 선택되는 것이 일반적이다.In addition to the above, the Astragalus extract or ethyl acetate fraction of the present invention is a flavor, such as various nutrients, vitamins, minerals (electrolytes), synthetic flavors and natural flavors, coloring and neutralizing agents (cheese, chocolate, etc.), pectic acid and its Salts, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated drinks and the like. In addition, the Astragalus extract or ethyl acetate fraction of the present invention may contain a flesh for preparing natural fruit juice and fruit juice beverage and vegetable beverage. These components can be used independently or in combination. The proportion of such additives is not so critical but is generally selected in the range of 0.1 to about 20 parts by weight per 100 parts by weight of the extract or ethyl acetate fraction of the present invention.
이하 본 발명을 실시예, 실험예 및 제제예에 의해 상세히 설명한다.Hereinafter, the present invention will be described in detail by Examples, Experimental Examples and Formulation Examples.
단, 하기 실시예, 실험예 및 제제예는 본 발명을 예시하는 것일 뿐, 본 발명의 내용이 하기 실시예, 실험예 및 제제예에 한정되는 것은 아니다.However, the following Examples, Experimental Examples, and Formulation Examples are merely illustrative of the present invention, and the content of the present invention is not limited to the following Examples, Experimental Examples, and Formulation Examples.
<< 실시예Example 1> 1> 황기Hwanggi 추출물의 제조 Preparation of Extract
<1-1> 70% 에탄올 이용한 <1-1> using 70% ethanol 황기Hwanggi 추출물 제조 Extract manufacturer
경동시장(서울)에서 구입한 황기를 건조한 후 400 g 시료의 중량의 10배에 해당하는 70% 에탄올 4 L를 첨가하여 6 시간 환류가열 추출한 후, 감압 여과하여 추출액과 잔사를 분리하였다. 이후 감압회전농축기를 사용하여 감압농축하여 각각의 농축액을 얻었으며, 이를 이용하여 -70℃에서 동결한 후 동결건조기를 이용하여 분말 98.52 g을 수득하였다. 이때 수율은 24.63%로 계산되었다. 최종 추출물을 “HP123"이라 명명하고 하기 실험의 시료로 사용하였다.After drying the sulfuric acid purchased from Gyeongdong market (Seoul), 4 L of 70% ethanol, which is 10 times the weight of 400 g of sample, was added thereto, and the mixture was heated under reflux for 6 hours, and then filtered under reduced pressure to separate the extract and the residue. Thereafter, the resultant was concentrated under reduced pressure using a vacuum rotary concentrator, to obtain respective concentrates, which were frozen at −70 ° C., and then obtained 98.52 g of a powder using a freeze dryer. Yield was calculated to be 24.63%. The final extract was named “HP123” and used as a sample for the following experiment.
<1-2> 100% 에탄올을 이용한 <1-2> using 100% ethanol 황기Hwanggi 추출물 제조 Extract manufacturer
경동시장(서울)에서 구입한 황기를 건조한 후 400 g 시료의 중량의 10배에 해당하는 100% 에탄올 4 L를 첨가하여 6 시간 환류가열 추출한 후, 감압 여과하여 추출액과 잔사를 분리하였다. 이후 감압회전농축기를 사용하여 감압농축하여 각각의 농축액을 얻었으며, 이를 이용하여 -70℃에서 동결한 후 동결건조기를 이용하여 분말 12 g을 수득하였다. After drying the sulfuric acid purchased from Gyeongdong market (Seoul), 4 L of 100% ethanol corresponding to 10 times the weight of the 400 g sample was added thereto, followed by extraction under reflux heating for 6 hours, followed by filtration under reduced pressure to separate the extract and the residue. Thereafter, the resultant was concentrated under reduced pressure using a vacuum rotary concentrator to obtain respective concentrates. After freezing at -70 ° C using this, 12 g of a powder was obtained using a lyophilizer.
<1-3> 증류수를 이용한 <1-3> using distilled water 황기Hwanggi 추출물 제조 Extract manufacturer
경동시장(서울)에서 구입한 황기를 건조한 후 400 g 시료의 중량의 10배에 해당하는 증류수 4 L를 첨가하여 6 시간 환류가열 추출한 후, 감압 여과하여 추출액과 잔사를 분리하였다. 이후 감압회전농축기를 사용하여 감압농축하여 각각의 농축액을 얻었으며, 이를 이용하여 -70℃에서 동결한 후 동결건조기를 이용하여 분말 9.4 g을 수득하였다. After drying the sulfuric acid purchased from Gyeongdong Market (Seoul), 4 L of distilled water corresponding to 10 times the weight of 400 g sample was added thereto, and the mixture was heated under reflux for 6 hours, and then filtered under reduced pressure to separate the extract and the residue. Thereafter, the resultant was concentrated under reduced pressure using a reduced pressure rotary concentrator to obtain respective concentrates. After freezing at -70 ° C using this, 9.4 g of a powder was obtained using a freeze dryer.
<1-4> 70% 메탄올을 이용한 <1-4> using 70% methanol 황기Hwanggi 추출물 제조 Extract manufacturer
경동시장(서울)에서 구입한 황기를 건조한 후 400 g 시료의 중량의 10배에 해당하는 70% 메탄올 4 L를 첨가하여 6 시간 환류가열 추출한 후, 감압 여과하여 추출액과 잔사를 분리하였다. 이후 감압회전농축기를 사용하여 감압농축하여 각각의 농축액을 얻었으며, 이를 이용하여 -70℃에서 동결한 후 동결건조기를 이용하여 분말 15 g을 수득하였다. After drying the Astragalus purchased from Gyeongdong Market (Seoul), 4 L of 70% methanol, which is 10 times the weight of 400 g of sample, was added thereto, and the mixture was heated under reflux for 6 hours, and then filtered under reduced pressure to separate the extract and the residue. Thereafter, the resultant was concentrated under reduced pressure using a vacuum rotary concentrator, to obtain respective concentrates. After freezing at -70 ° C using this, 15 g of a powder was obtained using a freeze dryer.
<1-5> 100% 메탄올을 이용한 <1-5> using 100% methanol 황기Hwanggi 추출물 제조 Extract manufacturer
경동시장(서울)에서 구입한 황기를 건조한 후 400 g 시료의 중량의 10배에 해당하는 70% 에탄올 4 L를 첨가하여 6 시간 환류가열 추출한 후, 감압 여과하여 추출액과 잔사를 분리하였다. 이후 감압회전농축기를 사용하여 감압농축하여 각각의 농축액을 얻었으며, 이를 이용하여 -70℃에서 동결한 후 동결건조기를 이용하여 분말 8.3 g을 수득하였다. After drying the sulfuric acid purchased from Gyeongdong market (Seoul), 4 L of 70% ethanol, which is 10 times the weight of 400 g of sample, was added thereto, and the mixture was heated under reflux for 6 hours, and then filtered under reduced pressure to separate the extract and the residue. Thereafter, the resultant was concentrated under reduced pressure using a reduced pressure rotary concentrator to obtain respective concentrates, which were frozen at −70 ° C., and then 8.3 g of powder was obtained using a freeze dryer.
<< 실시예Example 2> 2> 황기Hwanggi 추출물의 Of extract 분획물Fraction 제조 Produce
획득한 황기 추출분말 100 g을 1 L의 70% 에탄올에 현탁시킨 후 각각 1 L의 헥산, 에틸아세테이트, 물포화부탄올로 단계적으로 용매의 극성을 높여가면서 각 용매의 가용분획별로 3회씩 분획한 다음, 에틸아세테이트 분획층을 모아 회전 감압 농축기로 완전 농축하고 동결건조하였다. 그 결과 3.1 g의 에틸아세테이트 분획 분말을 얻을 수 있었다. 100 g of the obtained extract was extracted in 1 L of 70% ethanol, and then fractionated three times for each soluble fraction of each solvent while gradually increasing the polarity of the solvent with 1 L of hexane, ethyl acetate and water saturated butanol. The ethyl acetate fractionated layers were collected, concentrated completely with a rotary vacuum concentrator, and lyophilized. As a result, 3.1 g of ethyl acetate fraction powder was obtained.
<< 실험예Experimental Example 1> 1> 황기Hwanggi 추출물 및 이로부터 분리된 에틸아세테이트 Extract and ethyl acetate isolated therefrom 분획물에In fractions 의한 by 성장의 측정Measure of growth
황기 추출물에 의한 골길이 성장의 측정을 위하여, 실험에 이용한 동물은 스프라규-다울리(Sprague-Dawley)계 수컷 흰쥐(3주령)로 샘타코(대한민국)로부터 구입하여 사용하였고, 실험절차는 미 국립보건원의 동물관리 지침에 의해서 수행되었다. 온도는 20 ± 2℃, 조명은 07:00~19:00시 사이 조건으로 통일하였으며, 음식과 물은 리비툼(libitum)을 통해 공급되었으며, 쥐의 무게는 매일 측정되었다.For the measurement of bone length growth by Astragalus extract, the animals used in the experiment were Sprague-Dawley male rats (3 weeks old) purchased from Samtako (Korea), and the experimental procedure was used. This was done by the National Institutes of Health animal care guidelines. Temperatures were unified under conditions of 20 ± 2 ° C and illumination between 07:00 and 19:00 hours, food and water were supplied through libitum, and rats were weighed daily.
실험쥐들은 10마리씩 4개의 군으로 나누어, 음성 대조군에게는 식염수를, 실험군에게는 100 ㎎/㎏의 황기 70% 에탄올추출물(실험군 1) 및 500 ㎎/㎏의 황기 70% 에탄올 추출물(실험군 2), 100 mg/kg의 황기 100% 에탄올 추출물(실험군 3), 100 mg/kg 황기 70% 메탄올 추출물(실험군 4), 100 mg/kg 황기 100% 메탄올 추출물(실험군 5), 100 mg/kg 황기 증류수 추출물(실험군 6) 및 100 mg/kg 황기 에틸아 세테이트 분획물(실험군 7)을 매일 2회씩 4일간 경구 투여하였고, 양성 대조군에는 성장호르몬 (Eutropin, LG생명과학, Korea)을 20 ㎍/㎏ 농도로 피하 주사하였으며, 3일째에는 테트라사이클린(tetracycline; 10 ㎎/㎏)을 복강 주사하여 성장판에 침착되게 하였다. 테트라사이클린 주사로부터 48 시간이 경과한 5일째에는 흰쥐들을 에테르로 마취하여 희생하였다.The rats were divided into four groups of 10 rats, saline solution for the negative control group, 100 mg / kg Astragalus 70% ethanol extract (Experimental Group 1) and 500 mg / kg Astragalus 70% Ethanol extract (Experimental Group 2), 100 100 mg /
실험 5일째 희생한 흰쥐로부터 분리한 족경골은 4% 인산염-완충액 파라포름알데히드(phosphate-buffered paraformaldehyde)에 48 시간 고정시킨 후, 30% 수크로오스(sucrose) 용액에 침지시켜 탈수하였다. 이 후 고정된 골조직을 동결한 후 크라요컷(cryocut)을 사용하여 족경골(tibia) 근위부(proximal part)의 시상절편(sagital section)을 매 40 ㎛씩 절편한 후 수집하여 조직 표본으로 사용하였다. 그 후 절편은 길이 성장분석을 위해 사용되었다. On the fifth day of the experiment, the femur tibia isolated from the sacrificed rats was fixed in 4% phosphate-buffered paraformaldehyde for 48 hours and then immersed in 30% sucrose solution to dehydrate. Thereafter, frozen bone tissue was frozen and cryocut was used to cut the sagittal sections of the tibia proximal part every 40 μm and collected and used as tissue samples. Sections were then used for length growth analysis.
성장의 측정을 위해 성장판과 테트라사이클린(tetracycline)으로 침착되어 발색한 발광선과의 간격을 형광현미경으로 측정하였고(Hansson et al ., Calcified Tissue Research , 10(3), 238, 1972, 도 1), 각 절편마다의 평균값을 산출하였다. 모든 절차는 2명의 관찰자에 의해 맹검법으로 측정되었다. 결과는 평균 ± 표준편차로 표현되었으며, 모든 그룹간의 차이는 스튜던트-T 테스트(Student-T test) 검정법으로 하고 표준편차에서 유의한 차이가 있으나 검정이 적절치 않은 경우에는 크루스칼-왈리스 테스트(Kruskall-Wallis test) 검정법을 이용하였다. For the measurement of growth, the distance between the growth plate and the emission line developed by tetracycline was measured by fluorescence microscopy (Hansson et al. al ., Calcified Tissue Research , 10 (3), 238, 1972, Fig. 1), the average value for each section was calculated. All procedures were blinded by two observers. The results were expressed as mean ± standard deviation, and the difference between all groups was the Student-T test test and there was a significant difference in the standard deviation, but the Kruskall-Wallis test was not appropriate. Wallis test) was used.
그 결과, 황기 추출물 또는 에틸아세테이트 분획물의 투여는 음성 대조군보다 통계적으로 유의하게 성장을 가져왔으며 성장호르몬을 투여한 양성 대조군보다 도 우수한 골성장 효능을 보였다. 도 2는 표 3의 결과 중 음성 대조군, 양성 대조군, 실험군 1, 실험군 2의 성장을 나타낸 것이다. **는 P< 0.01을 나타낸다. As a result, administration of the Astragalus extract or ethyl acetate fractions showed statistically significant growth than the negative control and showed better bone growth efficacy than the positive control administered the growth hormone. Figure 2 shows the growth of the negative control group, positive control group, experimental group 1, experimental group 2 of the results in Table 3. ** indicates P <0.01.
또한 식염수를 투여한 음성 대조군의 성장을 100%로 하였을 경우, 황기 추출물(실험군 1 내지 실험군 6) 투여군, 황기 에틸아세테이트 분획물 투여군, 양성 대조군의 성장률을 계산하여 보았다. 그 결과, 역시 황기 추출물 투여군 및 에틸아세테이트의 성장률은 음성 대조군 보다 유의성 있게 높았으며, 양성 대조군보다도 높게 나타났다. In addition, when the growth rate of the negative control group administered with saline was 100%, the growth rate of the Astragalus extract (Experimental Group 1 to Experimental Group 6) administration group, Astragalus ethyl acetate fraction administration group, and positive control group was calculated. As a result, the growth rate of the Astragalus extract administration group and ethyl acetate was significantly higher than that of the negative control group and higher than that of the positive control group.
도 3은 표 1의 결과 중 음성 대조군, 양성 대조군, 실험군 1, 실험군 2의 성장률을 나타낸 것이다. **는 P< 0.01을 나타낸다. Figure 3 shows the growth rate of the negative control group, positive control group, experimental group 1, experimental group 2 of the results in Table 1. ** indicates P <0.01.
통계학적 검정 : student-T testStatistical test: student-T test
** : P < 0.01, *** : P < 0.001 ** : P <0.01, *** : P <0.001
1)계산식 = 100 + (실험군 길이성장 - 음성 대조군 길이성장)/음성 대조군 길이성장*100 1) Formula = 100 + (Experimental Length Growth-Negative Control Length Growth) / Negative Control Length Growth * 100
<실험예 2> 황기 추출물 및 에틸아세테이트 분획물의 급성 독성실험Experimental Example 2 Acute Toxicity Test of Astragalus Extract and Ethyl Acetate Fraction
황기 추출물 또는 에틸아세테이트 분획물에 대한 급성 독성을 알아보기 위하여, 하기와 같은 실험을 수행하였다.In order to determine the acute toxicity of the Astragalus extract or ethyl acetate fraction, the following experiment was performed.
4주령의 특정 병원체 부재(SPF, specific pathogens free) ICR계 마우스를 암수 각각 3 마리씩 4개군(암수 각각 3마리/실험군)으로 나누어, 온도 22 ± 3℃, 습도 55 ± 10%, 조명 12L/12D 조건의 동물실 내에서 사육하였다. 마우스는 실험에 사용되기 전 1주일 정도 순화시켰다. 실험 동물용 사료(마우스 및 랫트용, Dyets, 미국) 및 음수를 멸균한 후 공급하였으며 자유 섭취시켰다.SPF (specific pathogens free) ICR mice divided into 4 groups (3 males and 3 females / experimental group) at 4 weeks of age, temperature 22 ± 3 ° C, humidity 55 ± 10%, illumination 12L / 12D It was bred in a condition animal room. Mice were allowed to acclimate for about a week before being used in the experiment. Feed for experimental animals (for mice and rats, Dyets, USA) and negative water were supplied after sterilization and free intake.
상기 실시예에서 제조한 황기 70% 에탄올 추출물 및 에틸아세테이트 분획물을 0.5% 트윈(tween) 80에 50 ㎎/㎖ 농도로 조제한 후, 마우스 체중 20 g 당 0.04 ㎖(100 ㎎/㎏), 0.2 ㎖(500 ㎎/㎏) 및 0.4 ㎖(1,000 ㎎/㎏)씩 경구 투여하였다. 시료는 단회 경구 투여하였으며, 투여 후 7일 동안 다음과 같이 부작용 또는 치사 여부를 관찰하였다. 즉, 투여 당일은 투여 후 1 시간, 4 시간, 8 시간, 12 시간 뒤에, 그리고 투여 익일부터 7 일째까지는 매일 오전, 오후 1 회 이상씩 일반 증상의 변화 및 사망 동물의 유무를 관찰하였다. 또한, 투여 7 일째에 동물을 치사시켜 해부한 후 육안으로 내부 장기를 검사하였다. 투여 당일부터 1일 간격으로 체중의 변화를 측정하여 황기 추출물에 의한 동물의 체중 감소 현상을 관찰하였다.The 70% ethanol extract and ethyl acetate fractions prepared in Example were prepared at a concentration of 50 mg / ml in 0.5
실험 결과, 시료를 투여한 모든 마우스에서 특기할 만한 임상증상이 나타나지 않았고 폐사된 마우스도 없었으며, 또한 체중변화, 혈액검사, 혈액생화학 검사, 부검소견 등에서도 독성변화는 관찰되지 않았다.As a result, no significant clinical symptoms were observed in all the mice treated with the sample, no mice died, and no toxicity change was observed in weight change, blood test, blood biochemistry test, autopsy findings, etc.
그러므로 황기 70% 에탄올 추출물 및 에틸아세테이트 분획물은 모든 마우스에서 1,000 ㎎/㎏까지 독성변화를 나타내지 않았으며, 경구투여 최소치사량(LD50)이 1,000 ㎎/㎏ 이상인 안전한 물질로 판단되었다.Therefore, 70% ethanol extract and ethyl acetate fraction of Astragalus did not show toxicity change up to 1,000 mg / kg in all mice, and it was judged as a safe substance with oral administration minimum dose (LD 50 ) of 1,000 mg / kg or more.
하기에 본 발명의 조성물을 위한 제제예를 예시한다.Examples of formulations for the composition of the present invention are illustrated below.
<제제예 1> 약학적 제제의 제조Preparation Example 1 Preparation of Pharmaceutical Formulation
<1-1> 주사제제의 제조<1-1> Preparation of Injection
황기 70% 에탄올 추출물 10 ㎎Astragalus 70% Ethanol Extract 10 mg
소디움 메타비설파이트 3.0 ㎎Sodium Metabisulfite 3.0 mg
메틸파라벤 0.8 ㎎Methylparaben 0.8 mg
프로필파라벤 0.1 mgPropylparaben 0.1 mg
주사용 멸균증류수 적량Appropriate sterile distilled water for injection
상기의 성분을 혼합하고 통상의 방법으로 2 ㎖로 한 후, 2 ㎖ 용량의 앰플에 충전하고 멸균하여 주사제를 제조한다.The above ingredients are mixed and made into 2 ml by a conventional method, and then filled into 2 ml ampoules and sterilized to prepare an injection.
<1-2> 정제의 제조<1-2> Preparation of Tablet
황기 에틸에세테이트 분획물 200 ㎎Astragalus
유당 100 ㎎
전분 100 ㎎
스테아린산 마그네슘 적량Magnesium stearate proper amount
상기의 성분을 혼합하고 통상의 정제의 제조방법에 따라서 타정하여 정제를 제조한다.The above components are mixed and tableted according to a conventional method for producing tablets to produce tablets.
<1-3> 캡슐제의 제조<1-3> Preparation of Capsule
황기 70% 에탄올 추출물 10 ㎎Astragalus 70% Ethanol Extract 10 mg
유당 50 ㎎Lactose 50 mg
전분 50 ㎎Starch 50 mg
탈크 2 ㎎Talc 2 mg
스테아린산 마그네슘 2 ㎎2 mg magnesium stearate
상기의 성분을 혼합하고 통상의 캡슐제의 제조방법에 따라서 젤라틴 캡슐에 충전하여 캡슐제를 제조한다.Capsules are prepared by mixing the above ingredients and filling into gelatin capsules according to a conventional method for preparing capsules.
<1-4> <1-4> 액제의Liquid 제조 Produce
황기 에틀아세테이트 분획물 1000 ㎎Astragalus ethyl acetate fraction 1000 mg
설탕 20 g20 g of sugar
이성화당 20 g20 g of isomerized sugar
레몬향 적량Lemon flavor
정제수를 가하여 전체 1000 ㎖로 맞추었다. 통상의 액제의 제조방법에 따라 상기의 성분을 혼합한 다음, 갈색병에 충전하고 멸균시켜 액제를 제조한다.Purified water was added to adjust the total volume to 1000 ml. According to the conventional method for preparing a liquid, the above components are mixed, and then filled into a brown bottle and sterilized to prepare a liquid.
<< 제제예Formulation example 2> 식품의 제조 2> Manufacture of food
<2-1> 건강식품의 제조<2-1> Preparation of health food
황기 70% 에탄올 추출물 1000 ㎎Astragalus 70% Ethanol Extract 1000mg
비타민 혼합물 적량Vitamin mixture proper amount
비타민 A 아세테이트 70 ㎍70 μg of Vitamin A Acetate
비타민 E 1.0 ㎎Vitamin E 1.0 mg
비타민 B1 0.13 ㎎Vitamin B1 0.13 mg
비타민 B2 0.15 ㎎Vitamin B2 0.15 mg
비타민 B6 0.5 ㎎Vitamin B6 0.5 mg
비타민 B12 0.2 ㎍0.2 μg of vitamin B12
비타민 C 10 ㎎Vitamin C 10 mg
비오틴 10 ㎍10 μg biotin
니코틴산아미드 1.7 ㎎Nicotinic Acid 1.7 mg
엽산 50 ㎍50 μg folic acid
판토텐산 칼슘 0.5 ㎎Calcium Pantothenate 0.5mg
무기질 혼합물 적량Mineral mixture
황산제1철 1.75 ㎎Ferrous Sulfate 1.75 mg
산화아연 0.82 ㎎Zinc Oxide 0.82 mg
탄산마그네슘 25.3 ㎎Magnesium carbonate 25.3 mg
제1인산칼륨 15 ㎎Potassium monophosphate 15 mg
제2인산칼슘 55 ㎎Dibasic calcium phosphate 55 mg
구연산칼륨 90 ㎎
탄산칼슘 100 ㎎
염화마그네슘 24.8 ㎎Magnesium chloride 24.8 mg
상기의 비타민 및 미네랄 혼합물의 조성비는 비교적 건강식품에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 그 배합비를 임의로 변형 실시하여도 무방하며, 통상의 건강식품 제조방법에 따라 상기의 성분을 혼합한 다음, 통상의 방법에 따라 건강식품 조성물 제조(예, 영양캔디 등)에 사용할 수 있다.Although the composition ratio of the above-mentioned vitamin and mineral mixtures is mixed with a component suitable for a health food in a preferred embodiment, the compounding ratio may be arbitrarily modified, and the above ingredients are mixed according to a conventional health food manufacturing method. It can be used in the manufacture of health food compositions (eg nutritional candy, etc.) according to conventional methods.
<2-2> 밀가루 식품의 제조<2-2> Preparation of Flour Food
황기 에틸아세테이트 분획물 0.1 ~ 10.0 중량부를 밀가루에 첨가하고, 이 혼합물을 이용하여 통상의 방법으로 빵, 케이크, 쿠키, 크래커 및 면류를 제조하여 건강 증진용 식품을 제조하였다.0.1-10.0 parts by weight of the ethyl acetate acetate fraction was added to the flour, and bread, cake, cookies, crackers and noodles were prepared in a conventional manner using the mixture to prepare foods for health promotion.
<2-3> <2-3> 스프soup 및 육즙( And juicy ( graviesgravies )의 제조Manufacturing
황기 70% 에탄올 추출물 0.1 ~ 1.0 중량부를 스프 및 육즙에 첨가하여 통상의 방법으로 건강 증진용 육가공 제품, 면류의 수프 및 육즙을 제조하였다.Astragalus 70% ethanol extract 0.1 ~ 1.0 parts by weight was added to the soup and gravy to prepare a health-promoting meat products, soup and noodles of the noodles in a conventional manner.
<2-4> 그라운드 <2-4> ground 비프(ground beef)의Beef 제조 Produce
황기 70% 에탄올 추출물 10 중량부를 그라운드 비프에 첨가하여 통상의 방법으로 건강 증진용 그라운드 비프를 제조하였다.10 parts by weight of the 70% ethanol extract of Astragalus was added to ground beef to prepare ground beef for health promotion in a conventional manner.
<2-5> 유제품(<2-5> Dairy Products ( dairydairy productsproducts )의 제조Manufacturing
황기 70% 에탄올 추출물 0.1 ~ 1.0 중량부를 우유에 첨가하고, 상기 우유를 이용하여 통상의 방법으로 버터 및 아이스크림과 같은 다양한 유제품을 제조하였다.0.1-1.0 parts by weight of Astragalus 70% ethanol extract was added to milk, and various dairy products such as butter and ice cream were prepared in a conventional manner using the milk.
<2-6> <2-6> 선식의Solar 제조 Produce
현미, 보리, 찹쌀, 율무를 공지의 방법으로 알파화시켜 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.Brown rice, barley, glutinous rice, and yulmu were dried by a known method and dried, and the mixture was granulated to a powder having a particle size of 60 mesh.
검정콩, 검정깨, 들깨도 공지의 방법으로 쪄서 건조시킨 것을 배전한 후 분쇄기로 입도 60 메쉬의 분말로 제조하였다.Black soybeans, black sesame seeds, and perilla seeds were steamed and dried by a conventional method, and then they were prepared into powder having a particle size of 60 mesh by a pulverizer.
황기 70% 에탄올 추출물을 진공 농축기에서 감압, 농축하고, 분무, 열풍건조기로 건조하여 얻은 건조물을 분쇄기로 입도 60 메쉬로 분쇄하여 건조분말을 얻었다.Anhydrous 70% ethanol extract was concentrated under reduced pressure in a vacuum concentrator, dried by spraying and a hot air dryer, and ground to a particle size of 60 mesh using a grinder to obtain a dry powder.
상기에서 제조한 곡물류, 종실류 및 황기 추출물의 건조분말을 다음의 비율로 배합하여 통상의 방법으로 제조하였다.The dry powder of the grains, seeds and Astragalus extract prepared above was blended in the following ratio to prepare a conventional method.
곡물류(현미 30 중량부, 율무 15 중량부, 보리 20 중량부),Cereals (30 parts by weight brown rice, 15 parts by weight brittle, 20 parts by weight of barley),
종실류(들깨 7 중량부, 검정콩 8 중량부, 검정깨 7 중량부),Seeds (7 parts by weight of perilla, 8 parts by weight of black beans, 7 parts by weight of black sesame seeds)
황기 추출물의 건조분말(1 중량부),Dry powder (1 part by weight) of Astragalus extract,
영지(0.5 중량부),(0.5 part by weight),
지황(0.5 중량부)Foxglove (0.5 part by weight)
<< 제제예Formulation example 3> 음료의 제조 3> Manufacturing of beverage
<3-1> 건강 음료의 제조<3-1> Preparation of Healthy Drinks
황기 70% 에탄올 추출물 1000 ㎎Astragalus 70% Ethanol Extract 1000mg
구연산 1000 ㎎Citric acid 1000 mg
올리고당 100 g100 g oligosaccharides
매실농축액 2 gPlum concentrate 2 g
타우린 1 g1 g of taurine
정제수를 가하여 전체 900 ㎖Add 900 ml of purified water
통상의 건강음료 제조방법에 따라 상기의 성분을 혼합한 다음, 약 1시간 동안 85℃에서 교반 가열한 후, 만들어진 용액을 여과하여 멸균된 2 ℓ용기에 취득하여 밀봉 멸균한 뒤 냉장 보관한 다음 본 발명의 건강음료 조성물 제조에 사용한다. After mixing the above components according to the conventional healthy beverage production method, and stirred and heated at 85 ℃ for about 1 hour, the resulting solution is filtered and obtained in a sterilized 2 L container, sealed sterilization and then refrigerated and stored Used to prepare the healthy beverage composition of the invention.
상기 조성비는 비교적 기호음료에 적합한 성분을 바람직한 실시예로 혼합 조성하였지만, 수요계층, 수요국가, 사용 용도 등 지역적, 민족적 기호도에 따라서 그 배합비를 임의로 변형 실시하여도 무방하다.Although the composition ratio is mixed with a relatively suitable component in a preferred embodiment in a preferred embodiment, the compounding ratio may be arbitrarily modified according to regional and ethnic preferences such as demand hierarchy, demand country, use purpose.
<3-2> <3-2> 야채쥬스의Vegetable juice 제조 Produce
황기 70% 에탄올 추출물 0.5 g을 토마토 또는 당근 등의 야채의 쥬스 1,000 ㎖에 가하여 통상의 방법으로 건강 증진용 야채쥬스를 제조하였다.0.5 g of 70% ethanol extract of Astragalus was added to 1,000 ml of a vegetable juice such as tomato or carrot to prepare a vegetable juice for health promotion in a conventional manner.
<3-3> <3-3> 과일쥬스의Fruit juice 제조 Produce
황기 에틸아세테이트 분획물 0.1 g을 사과 또는 포도 등의 과일의 쥬스 1,000 ㎖에 가하여 통상의 방법으로 건강 증진용 과일쥬스를 제조하였다.0.1 g of Astragalus ethyl acetate fraction was added to 1,000 ml of fruit juice, such as apple or grape, to prepare a fruit juice for health promotion in a conventional manner.
도 1은 흰쥐에 테트라사이클린(tetracycline) 주사 후, 침착된 후지경골 골단부 성장판에 침착되어 발생하는 발광을 형광현미경으로 촬영한 사진이다: Figure 1 is a photograph taken by fluorescence microscopy of luminescence resulting from deposition on the deposited tibial epiphyseal growth plate after tetracycline injection in rats:
A: 식염수 투여군; B: 성장호르몬(20 ㎍/㎏) 투여군; 및 C: 황기 추출물(HP123, 100 ㎎/㎏) 투여군, D: 황기 추출물(HP123, 500 ㎎/㎏) 투여군A: saline administration group; B: growth hormone (20 µg / kg) administration group; And C: Astragalus extract (HP123, 100 mg / kg) administration group, D: Astragalus extract (HP123, 500 mg / kg) administration group
도 2는 흰쥐에 테트라사이클린 주사한 뒤 48시간 후, 성장판과 발광선과의 간격을 측정한 그래프이다: Figure 2 is a graph measuring the interval between the growth plate and the luminescent line 48 hours after tetracycline injection in rats:
**: P< 0.01, ***: P< 0.001. ** : P <0.01, *** : P <0.001.
도 3은 흰쥐에서의 성장률을 나타낸 그래프이다:3 is a graph showing growth rate in rats:
**: P< 0.01, ***: P< 0.001. ** : P <0.01, *** : P <0.001.
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