KR20190142025A - Pharmaceutical composition and food composition for improving the symptoms of menopausal symptoms - Google Patents
Pharmaceutical composition and food composition for improving the symptoms of menopausal symptoms Download PDFInfo
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- KR20190142025A KR20190142025A KR1020180069047A KR20180069047A KR20190142025A KR 20190142025 A KR20190142025 A KR 20190142025A KR 1020180069047 A KR1020180069047 A KR 1020180069047A KR 20180069047 A KR20180069047 A KR 20180069047A KR 20190142025 A KR20190142025 A KR 20190142025A
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Abstract
Description
본 발명은 갱년기 증상 개선용 약학적 조성물에 관한 것으로, 보다 상세하게는 다양한 생약을 이용하여 제조된 갱년기 증상 개선용 약학적 조성물에 관한 것이다.The present invention relates to a pharmaceutical composition for improving menopausal symptoms, and more particularly, to a pharmaceutical composition for improving menopausal symptoms manufactured using various herbal medicines.
내분비계 호르몬의 하나인 에스트로겐은 체내에서 콜레스테롤로부터 합성되며, 분비원은 주로 난소의 여포 및 황체이다. 뇌하수체전엽에서 분비되는 성선자극호르몬(gonadotropin)이 생식기관에 신호를 전달하면 월경주기를 조절하고 임신을 가능케 하는 호르몬인 여포자극호르몬(follicle stimulating hormone; FSH)과 황체형성호르몬(Lutenizing hormone; LH)이 분비되고, 이는 다시 난소에 작용하여 에스트로겐과 프로게스테론을 분비하게 한다. 상기 에스트로겐은 에스트라디올, 에스트론, 에스트리올 등을 총칭한다.Estrogen, one of endocrine hormones, is synthesized from cholesterol in the body, and the secretory sources are mainly follicles and corpus luteum of the ovaries. When the gonadotropin secreted by the anterior pituitary gland transmits a signal to the reproductive organs, follicle stimulating hormone (FSH) and luteenizing hormone (LH) are hormones that regulate the menstrual cycle and enable pregnancy Is secreted, which in turn acts on the ovary, releasing estrogen and progesterone. The estrogens collectively include estradiol, estrone, estriol and the like.
폐경 전의 여성에 있어 난소는 에스트로겐의 주요 공급원이다. 여성이 폐경기에 접어들면 난소가 노화됨에 따라 하수체성 성선자극 호르몬(여포자극 호르몬 및 황체형성 호르몬)에 대한 반응이 감소하고, 이로 인해 초기 여포기가 더욱 단축되어 월경 주기가 더욱 단축되며, 배란기가 더욱 단축되고, 프로게스테론 생성이 감소되며, 사이클이 더욱 불규칙하게 된다. 결국, 여포가 반응하지 못하게 되어 에스트로겐을 생성하지 못하게 된다. 이러한 에스트로겐의 분비 감소는 여성의 비뇨생식계통 뿐만 아니라 신체 전반에 걸쳐 큰 영향을 미치게 되며, 배란이 중단되어 월경이 사라지는 폐경 또는 그 이전에 호르몬의 감소로 인한 폐경기 증상이 나타나게 된다.In premenopausal women, the ovary is a major source of estrogen. As women enter menopause, as the ovary ages, the response to pituitary gonadotropins (follicle stimulating hormones and luteinizing hormones) decreases, which further shortens the early follicular phase, further shortening the menstrual cycle, It is shorter, the production of progesterone is reduced, and the cycle becomes more irregular. Eventually, the follicles will not react and will not produce estrogens. This decrease in estrogen secretion has a great effect not only on the genitourinary system of women but also throughout the body, and menopausal symptoms due to hormone reduction before or after menopause is stopped and menstruation disappears.
폐경기 이후 여성에서는 에스트로겐의 급속한 감소로 인하여, 심리학적 및 감성적 징후, 예컨대 피로, 흥분, 불면, 집중력 저하, 우울증, 기억 상실, 두통, 근심 및 신경과민이나 소심증 등이 발생할 수 있고, 재발성 안면 홍조에 의한 수면 방해에 따른 피로와 흥분, 간헐성 현기증, 감각이상, 심계항진 및 빈맥, 메스꺼움, 변비, 설사, 관절통, 근육통, 수족 냉증 및 체중 증가 현상의 발생우려 또한, 현저히 높아지게 된다.In postmenopausal women, rapid decreases in estrogen can lead to psychological and emotional signs such as fatigue, excitement, insomnia, poor concentration, depression, memory loss, headache, anxiety, and nervousness or timidity, and recurrent hot flashes. Fatigue and excitement due to sleep disturbance, intermittent dizziness, paresthesia, palpitations and tachycardia, nausea, constipation, diarrhea, joint pain, myalgia, cold hands and feet, and weight gain are also significantly increased.
또한, 골다공증이 쉽게 유발될 뿐 아니라, 심장병, 고혈압, 뇌졸중 등의 심혈관계 질환으로 인한 사망률도 증가한다(Kalin MF & Zumoff B. Steroids 55:330-352, 1990). In addition, osteoporosis is not only easily induced, but also mortality due to cardiovascular diseases such as heart disease, high blood pressure and stroke is increased (Kalin MF & Zumoff B. Steroids 55: 330-352, 1990).
또한, 피부 및 비뇨생식계통의 변화를 초래하며, 자가면역성 질환, 백내장 및 대장암 등의 발병 확률이 높아진다.In addition, it causes changes in the skin and urogenital system, and increases the probability of developing autoimmune diseases, cataracts and colorectal cancer.
현재, 상기 안면홍조 등 갱년기 증상을 개선하는데 있어서 가장 효과적인 공지의 방법은 에스트로겐을 투여하여 결핍된 에스트로겐을 보충해 주는 것으로 알려져 있다. 그러나, 합성 에스트로겐을 장기간에 걸쳐 투여하는 경우에는 유방암과 자궁암을 유발할 수 있다는 심각한 문제점이 있다(Tamini RT et al. Arch intern med 166:1483-1489, 2006, Gertig DM. Menopause 13:178-184, 2006, Strom BL et al. Am J Epidemiol 164:775-586, 2006, AraJr NL & Athanazio DA. Cad Saude Publica 23:2613-2622, 2007 참조). Currently, the most effective known method for improving menopausal symptoms such as hot flashes is known to supplement estrogens by administering estrogens. However, there is a serious problem that long-term administration of synthetic estrogens can lead to breast and uterine cancer (Tamini RT et al. Arch intern med 166: 1483-1489, 2006, Gertig DM. Menopause 13: 178-184, 2006, Strom BL et al. Am J Epidemiol 164: 775-586, 2006, AraJr NL & Athanazio DA.Cad Saude Publica 23: 2613-2622, 2007).
또한, 최근 들어서는 장기간에 걸친 합성 에스트로겐 대체 요법이 유방암을 유발할 가능성이 높다는 믿을만한 주장이 다수 제기되면서 학자들 간에는 물론, 사회 전반에 걸쳐 논란이 가열되고 있다. 아직 확실히 증명된 것은 아니지만, 합성 에스트로겐 대체 요법을 받은 여성에 있어서 유방암 발생 위험은 시간 경과에 따른 에스트로겐 노출 용량과 관련이 있는 것으로 믿어지고 있다.In recent years, controversy has been heating up among scholars and across society, with a number of credible claims that long-term synthetic estrogen replacement therapy is likely to cause breast cancer. Although not yet proven, it is believed that the risk of developing breast cancer in women receiving synthetic estrogen replacement therapy is associated with the dose of estrogen exposure over time.
따라서, 상기 호르몬 대체 요법에 사용되는 합성 에스트로겐보다 안전한 대체물로서 식물성 에스트로겐인 피토에스트로겐을 찾고자 하는 연구가 최근 활발히 진행되고 있다. 상기 피토에스트로겐은 동물의 체내에서 에스트로겐성 효과를 발휘하는, 식물 중에 존재하는 비스테로이드성 화합물을 지칭하며, 항암 작용과 심장병에 효과가 있는 것으로 알려져 있는 곡물, 과일 및 채소들은 대부분 피토에스트로겐을 미량 함유하고 있다.Therefore, researches to find phytoestrogens, phytoestrogens, as safer substitutes than synthetic estrogens used in the hormone replacement therapy have been actively conducted in recent years. The phytoestrogens refer to nonsteroidal compounds present in plants that exert an estrogenous effect in the body of animals, and most grains, fruits and vegetables known to be effective against anticancer activity and heart disease contain trace amounts of phytoestrogens. Doing.
등록특허 제10-1272602호에는 여성의 갱년기 증상을 완화하기 위하여 석류추출물을 이용한 방법이 기재되어 있고, 등록특허 제10-0486365호에는 대두, 홍삼, 당귀, 작약, 감초 및 비타민을 이용하여 갱년기 증상을 완화하는 조성물에 관하여 기재되어 있다.Patent No. 10-1272602 describes a method using pomegranate extract to alleviate menopausal symptoms in women, and Patent No. 10-0486365 uses soybean, red ginseng, Angelica, peony, licorice and vitamins to menopausal symptoms. Described are compositions that mitigate.
즉, 여성의 갱년기 증상을 개선하는데 효과적이면서도, 유방암, 자궁암 등을 유발하지 않는 등 안전한 약품 또는 건강 기능식품에 대한 필요성이 꾸준히 제기되어 오고 있다.In other words, there is a need for a safe drug or functional food that is effective in improving women's menopausal symptoms and does not cause breast cancer or uterine cancer.
따라서, 본 발명은 상기 문제를 해결하기 위해 안출된 것으로, 합성에스트로겐이 아닌 식물에서 추출된 추출액을 유효성분으로 포함하여 유방암, 자궁암 등을 유발하지 않는 등 갱년기 증상을 치료, 예방 및 개선하는데 효과적인 약학적 조성물을 제공하고자 한다.Therefore, the present invention has been made to solve the above problems, including the extract extracted from plants other than synthetic estrogen as an active ingredient effective in treating, preventing and improving menopausal symptoms such as not causing breast cancer, uterine cancer, etc. To provide a suitable composition.
본 발명의 다른 목적은 갱년기 증상을 치료, 예방 및 개선하는데 효과적인 식품 조성물을 제공하고자 한다.Another object of the present invention is to provide a food composition effective for treating, preventing and ameliorating menopausal symptoms.
상기 문제를 해결하기 위하여 본 발명은 길초근, 당귀, 천궁 및 작약의 혼합 추출물을 유효성분으로 포함하는 갱년기 증상 치료, 예방 및 개선용 약학적 조성물을 제공한다.In order to solve the above problems, the present invention provides a pharmaceutical composition for treating, preventing and improving menopausal symptoms, which comprises a mixed extract of Gilcho root, Angelica, Cervix and Peony as an active ingredient.
상기 약학적 조성물은 길초근 3 내지 18 중량%, 당귀 40 내지 60중량%, 천궁 10 내지 30중량%, 작약 10 내지 30중량%인 것을 특징으로 하는 약학적 조성물을 제공한다.The pharmaceutical composition provides a pharmaceutical composition, characterized in that 3 to 18% by weight of the root herb, Angelica 40 to 60% by weight, 10 to 30% by weight, 10 to 30% by weight peony.
상기 혼합 추출물은 물, 탄소수 1 내지 6인 알코올, 헥산, 에틸아세테이트 및 이들의 혼합용매로 이루어진 군에서 선택된 1종 이상의 용매로 추출한 것임을 특징으로 하는 조성물을 제공한다.The mixed extract provides a composition characterized in that extracted with one or more solvents selected from the group consisting of water, alcohol having 1 to 6 carbon atoms, hexane, ethyl acetate and a mixed solvent thereof.
상기 알코올은 에탄올인 것을 특징으로 하는 약학적 조성물을 제공한다.The alcohol provides a pharmaceutical composition, characterized in that ethanol.
한편, 길초근, 당귀, 천궁 및 작약의 혼합 추출물을 유효성분으로 포함하는 갱년기 증상 개선, 치료 및 면역효능을 갖는 식품 조성물을 제공한다.On the other hand, it provides a food composition having a menopausal symptom improvement, treatment and immune efficacy comprising a mixed extract of Gilcho root, Angelica, horoscope and Peony as an active ingredient.
본 발명에 따르면, 길초근, 당귀, 천궁 및 작약의 비율을 조절함으로써 합성에스트로겐을 대체하여 갱년기 증상을 개선, 예방 및 치료할 수 있는 약학적 조성물 및 식품 조성물을 제공할 수 있다. According to the present invention, it is possible to provide a pharmaceutical composition and a food composition that can improve, prevent and treat menopausal symptoms by replacing synthetic estrogens by controlling the ratio of gilcho root, angui, cheongung and peony.
도 1은 실시예 및 비교예에 따른 물질 투여 후 에스트로겐 활성을 측정하여 나타낸 그래프,
도 2는 실시예 및 비교예에 따른 물질 투여 후 에스트로겐 수용체 발현을 측정하여 나타낸 그래프,
도 3은 실시예 및 비교예에 따른 물질 투여 후 ALP 변화량을 측정한 그래프,
도 4는 실시예 및 비교예에 따른 물질 투여 후 혈중 에스트로겐 농도를 측정한 그래프,
도 5는 실시예 및 비교예에 따른 물질 투여 후 혈중 LH 및 FSH의 변화를 나타낸 그래프,
도 6은 실시예 및 비교예에 따른 물질 투여 후 자궁무게를 나타낸 그래프 및 자궁두께를 광학 현미경으로 관찰한 사진.1 is a graph showing the measurement of estrogen activity after administration of a substance according to Examples and Comparative Examples,
2 is a graph showing the measurement of estrogen receptor expression after administration of a substance according to Examples and Comparative Examples,
3 is a graph measuring the amount of ALP change after administration of a substance according to Examples and Comparative Examples;
4 is a graph measuring the concentration of estrogen in blood after administration of a substance according to Examples and Comparative Examples;
5 is a graph showing changes in blood LH and FSH after administration of a substance according to Examples and Comparative Examples;
Figure 6 is a graph showing the uterine weight and the thickness of the uterus after the administration of the material according to Examples and Comparative Examples photographed by optical microscope.
이하 바람직한 실시예를 통하여 본 발명을 상세히 설명하기로 한다. 이에 앞서, 본 명세서 및 청구범위에 사용된 용어나 단어는 통상적이거나 사전적인 의미로 한정해서 해석되어서는 아니 되며, 발명자는 그 자신의 발명을 가장 최선의 방법으로 설명하기 위해 용어의 개념을 적절하게 정의할 수 있다는 원칙에 입각하여, 본 발명의 기술적 사상에 부합하는 의미와 개념으로 해석되어야만 한다. 따라서, 본 명세서에 기재된 실시예의 구성은 본 발명의 가장 바람직한 일실시예에 불과할 뿐이고 본 발명의 기술적 사상을 모두 대변하는 것은 아니므로, 본 출원시점에 있어서 이들을 대체할 수 있는 다양한 균등물과 변형예들이 있을 수 있음을 이해하여야 한다. 또한, 명세서 전체에서, 어떤 부분이 어떤 구성요소를 "포함"한다고 할 때, 이는 특별히 반대되는 기재가 없는 한, 다른 구성요소를 제외하는 것이 아니라 다른 구성요소를 더 포함할 수 있음을 의미한다.Hereinafter, the present invention will be described in detail with reference to preferred embodiments. Prior to this, terms or words used in the present specification and claims should not be construed as being limited to the common or dictionary meanings, and the inventors should properly explain the concept of terms in order to best explain their own invention. Based on the principle that it can be defined, it should be interpreted as meaning and concept corresponding to the technical idea of the present invention. Therefore, the configuration of the embodiments described herein is only one of the most preferred embodiments of the present invention and does not represent all of the technical idea of the present invention, various equivalents and modifications that can replace them at the time of the present application It should be understood that there may be In addition, throughout the specification, when a part is said to "include" a certain component, it means that it may further include other components, not to exclude other components, unless otherwise stated.
본 발명자들은 갱년기 증상을 개선, 예방 및 치료할 수 있는 물질을 발굴하기 위해 연구를 거듭한 결과, 길초근, 당귀, 천궁 및 작약의 혼합추출물을 포함하는 약학 조성물이 갱년기 증상을 개선하는 호르몬인 에스트로겐과 유사한 효능이 있는 것을 발견하고, 상기 생약들의 비율 및 제조과정을 통해 현저한 효과를 낼 수 있도록 연구를 거듭한 결과 본 발명에 이르게 되었다.The present inventors have conducted research to find a substance that can improve, prevent and treat menopausal symptoms, and as a result, the pharmaceutical composition comprising a mixed extract of Gilcho root, Angelica, Cervix, and Peony, estrogen, a hormone that improves menopausal symptoms, As a result of discovering similar efficacy, the study was carried out to produce a remarkable effect through the ratio and manufacturing process of the herbal medicines, and thus led to the present invention.
따라서 본 발명은 길초근, 당귀, 천궁 및 작약의 혼합 추출물을 유효성분으로 포함하는 갱년기 증상 치료, 예방 및 개선용 약학적 조성물을 개시한다.Therefore, the present invention discloses a pharmaceutical composition for treating, preventing and improving menopausal symptoms, which comprises a mixed extract of Gilcho root, Angelica, Cervix and Peony as an active ingredient.
상기 길초근은 마타리과의 쥐오줌풀(Valeriana fauriei Briquet) 또는 동속 근연식물의 뿌리 및 뿌리줄기로 만든 약재로서, 정신불안으로 인한 가슴 뜀, 불면, 쉽게 놀라는 증상, 신경쇠약, 심근염, 산후 심장병, 심박쇠약, 고산병, 생리불순, 위경련, 관절점, 타박상 등에 사용된다고 알려져 있다.The herbaceous root is a medicinal herb made from the roots and rhizomes of Valerian fauriei Briquet of the Matariaceae family, or related plants, heart jump due to anxiety, blown away, easily surprised symptoms, nervous breakdown, myocarditis, postpartum heart disease, heart rate It is known to be used for weakness, altitude sickness, menstrual irregularities, stomach cramps, joint points, and bruises.
또한, 상기 당귀는 대표적인 약용 식물로서, 피가 부족할 때 피를 생성해 주는 보혈 작용에 효능이 있다고 알려져 있다.In addition, the Angelica is a representative medicinal plant, it is known that the effect on the blood production to produce blood when the blood is scarce.
또한, 상기 천궁은 약용식물로 쌍떡잎식물 이판화군 산형화목 미나리과의 여러해살이풀이다. 천궁은 진정, 진통, 강장 등에 효능이 있어 두통, 빈혈증, 부인병 등을 치료하는데 효능이 있다고 알려져 있다.In addition, the cheongung is a perennial herb of the dicotyledon plant cultivars apiaceae as a medicinal plant. Cheongung is known to be effective in treating headaches, anemia and gynecological diseases because it is effective in calming, pain and tonic.
또한, 상기 작약은 약용식물로 쌍떡잎식물 작약과 작약속의 여러해살이풀이다. 작약의 뿌리는 진통, 복통, 월경통, 무월경, 토혈, 빈혈, 타박상 등에 효능이 있다고 알려져 있다. In addition, the peony is a perennial herb of the dicotyledon and peony genus as a medicinal plant. Peony root is known to be effective in analgesic, abdominal pain, dysmenorrhea, amenorrhea, hemostasis, anemia, bruises.
본 발명에서 상기 약학적 조성물은 길초근이 3 내지 18중량%, 바람직하게는 5 내지 15중량%, 더욱 바람직하게는 8 내지 12중량%일 수 있고, 당귀는 40 내지 60중량%, 45 내지 55중량%, 더욱 바람직하게는 48 내지 52중량%일 수 있고, 천궁이 10 내지 30중량%, 바람직하게는 15 내지 25중량%, 더욱 바람직하게는 18 내지 22중량%일 수 있고, 작약이 10 내지 30중량%, 바람직하게는 15 내지 25중량%, 더욱 바람직하게는 18 내지 22중량%일 수 있다.In the present invention, the pharmaceutical composition may be 3 to 18% by weight, preferably 5 to 15% by weight, more preferably 8 to 12% by weight, guinea root is 40 to 60% by weight, 45 to 55 Weight percent, more preferably 48 to 52 weight percent, crossbow may be 10 to 30 weight percent, preferably 15 to 25 weight percent, more preferably 18 to 22 weight percent,
상기 길초근, 당귀, 천궁 및 작약의 용량이 상기 비율을 벗어난 경우, 갱년기 증상을 개선하는 효능이 낮아질 수 있다. When the doses of the herbaceous muscle, Angelica, horoscope and peony are out of the ratio, the efficacy of improving menopausal symptoms may be lowered.
본 발명에서의 혼합 추출물은 물, 탄소수 1 내지 6인 알코올, 헥산, 에틸아세테이트 및 이들의 혼합용매로 이루어진 군에서 선택된 1종 이상의 용매로 추출할 수 있다. 또한, 본 발명의 추출물은 바람직하게는 물, 탄소수 1 내지 6인 알코올 및 이들의 혼합용매에서 선택된 용매로 추출한 것일 수 있으며, 또는 탄소수 1 내지 6의 저급 알코올 수용액일 수 있다. 상기 알코올 수용액의 농도는 10 내지 100%(v/v)일 수 있으며, 바람직하게는 20 내지 50%(v/v)일 수 있다. 본 발명의 추출 용매는 가장 바람직하게는 물 또는 30%(v/v)의 에탄올 수용액일 수 있다. The mixed extract in the present invention can be extracted with one or more solvents selected from the group consisting of water, alcohols having 1 to 6 carbon atoms, hexane, ethyl acetate and mixed solvents thereof. In addition, the extract of the present invention may be preferably extracted with a solvent selected from water, alcohol having 1 to 6 carbon atoms and a mixed solvent thereof, or may be an aqueous solution of lower alcohol having 1 to 6 carbon atoms. The concentration of the aqueous alcohol solution may be 10 to 100% (v / v), preferably 20 to 50% (v / v). The extraction solvent of the present invention may be most preferably water or 30% (v / v) aqueous ethanol solution.
본 발명에서 상기 약학적 조성물은 하기와 같은 제조공정으로 제조될 수 있다.In the present invention, the pharmaceutical composition may be prepared by the following manufacturing process.
길초근, 당귀, 천궁 및 작약을 통상적인 추출방법에 따라 제조할 수 있고, 본 발명에서의 추출 방법은 당업계에 공지된 것이면 제한 없이 사용할 수 있으며, 그 예로 냉침, 열수 추출, 초음파 추출, 환류냉각 추출 등의 방법이 있으나 여기에 국한되는 것은 아니며, 바람직하게는 열수 추출할 수 있다. 예를 들어, 길초근, 당귀, 천궁 및 작약의 혼합물의 중량의 1 내지 20배, 바람직하게는 5 내지 15배 부피의 물, 탄소수 1 내지 6인 알코올, 헥산, 에틸아세테이트 및 이들의 혼합용매로 이루어진 군에서 선택된 1종 이상의 용매, 바람직하게는 30%(v/v)에탄올 수용액을 가하여 80 내지 95℃, 바람직하게는 85 내지 90℃에서 1 내지 5시간, 바람직하게는 2 내지 4시간 동안, 1 내지 5회, 바람직하게는 1 내지 3회 반복하여 추출할 수 있다. 상기 방법으로 제조된 추출액을 여지로 감압 여과한 다음, 여과액을 40 내지 80℃, 바람직하게는 40 내지 60℃에서 감압 농축 및 건조한 후 본 발명의 혼합 추출물을 수득할 수 있다.Gilchogeun root, Angelica, cheongunggi and peony can be prepared according to a conventional extraction method, the extraction method in the present invention can be used without limitation, if known in the art, for example, cold soak, hot water extraction, ultrasonic extraction, reflux There is a method such as cooling extraction, but is not limited thereto, and preferably hot water extraction. For example, with 1-20 times, preferably 5-15 times, volume of water, alcohol having 1-6 carbon atoms, hexane, ethyl acetate, and mixed solvents thereof, of the mixture of Geuncho root, Angelica, Cervix and Peony At least one solvent selected from the group consisting of, preferably 30% (v / v) ethanol aqueous solution, is added at 80 to 95 ℃, preferably 85 to 90 ℃ for 1 to 5 hours, preferably 2 to 4 hours, The extraction may be repeated 1 to 5 times, preferably 1 to 3 times. The extract prepared by the above method is filtered under reduced pressure, and then the filtrate is concentrated under reduced pressure and dried at 40 to 80 ° C, preferably 40 to 60 ° C, to obtain the mixed extract of the present invention.
본 발명에서 상기 혼합추출물을 포함하는 약학적 조성물은 길초근, 당귀, 천궁 및 작약의 단독 추출물을 혼합한 것일 수도 있고, 또는 길초근, 당귀, 천궁 및 작약을 혼합한 다음 추출한 복합 추출물일 수도 있다. 본 명세서에서 상기 단독 추출물의 혼합물 또는 복합 추출물을 모두 혼합 추출물로 통칭할 수 있다. 본 발명의 조성물은 유효성분으로 포함하는 길초근, 당귀, 천궁 및 작약 추출물의 혼합 방식에 제한받지 아니한다. 상기 추출물의 형태나 성상에는 제한이 없으며, 용액, 농축물일 수도 있고, 추출물 제조에 사용된 용매를 제거한 고형분 또는 분말일 수도 있다. In the present invention, the pharmaceutical composition comprising the mixed extract may be a mixture of the extracts of Gilcho root, Angelica, Cheongung and Peony, or may be a complex extract extracted after mixing Gilcho root, Angelica, Cheongung and Peony. . In the present specification, a mixture or a complex extract of the single extract may be collectively referred to as a mixed extract. The composition of the present invention is not limited to the mixing mode of the gilcho root, Angelica, Cheongung and Peony extracts included as an active ingredient. There is no restriction on the form or property of the extract, and may be a solution, a concentrate, or may be a solid or powder from which the solvent used for preparing the extract is removed.
상기 조성물이 약학적 조성물의 형태로 제공이 되는 경우에는, 본 발명의 상기 혼합물 이외에 하나 이상의 약학적으로 허용되는 담체, 부형제 또는 희석제를 추가로 함유할 수 있다. 상기에서 "약학적으로 허용되는"이란 생리학적으로 허용되고 인간에게 투여될 때, 활성성분의 작용을 저해하지 않으며 통상적으로 위장 장애, 현기증과 같은 알레르기 반응 또는 이와 유사한 반응을 일으키지 않는 비독성의 조성물을 말한다. When the composition is provided in the form of a pharmaceutical composition, it may further contain one or more pharmaceutically acceptable carriers, excipients or diluents in addition to the mixture of the present invention. As used herein, "pharmaceutically acceptable" means a non-toxic composition that, when administered to humans, does not inhibit the action of the active ingredient and usually does not cause an allergic reaction such as gastrointestinal disorders, dizziness, or the like. Say
약학적으로 허용되는 담체로는 예컨대, 경구 투여용 담체 또는 비경구 투여용 담체를 추가로 포함할 수 있다. 경구 투여용 담체는 락토스, 전분, 셀룰로스 유도체, 마그네슘 스테아레이트, 스테아르산 등을 포함할 수 있다. 아울러, 펩티드 제제에 대한 경구투여용으로 사용되는 다양한 약물전달물질을 포함할 수 있다. 또한, 비경구 투여용 담체는 물, 적합한 오일, 식염수, 수성 글루코오스 및 글리콜 등을 포함할 수 있으며, 안정화제 및 보존제를 추가로 포함할 수 있다. 적합한 안정화제로는 아황산수소나트륨, 아황산나트륨 또는 아스코르브산과 같은 항산화제가 있다. 적합한 보존제로는 벤즈알코늄 클로라이드, 메틸- 또는 프로필-파라벤 및 클로로부탄올이 있다. 본 발명의 약학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현택제 등을 추가로 포함할 수 있다. 그 밖의 약학적으로 허용되는 담체 및 제제는 다음의 문헌에 기재되어 있는 것을 참고로 할 수 있다(Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995).Pharmaceutically acceptable carriers may further include, for example, carriers for oral administration or carriers for parenteral administration. Carriers for oral administration may include lactose, starch, cellulose derivatives, magnesium stearate, stearic acid and the like. In addition, it may include a variety of drug delivery materials used for oral administration to the peptide formulation. In addition, carriers for parenteral administration may include water, suitable oils, saline, aqueous glucose and glycols, and the like, and may further include stabilizers and preservatives. Suitable stabilizers include antioxidants such as sodium hydrogen sulfite, sodium sulfite or ascorbic acid. Suitable preservatives include benzalkonium chloride, methyl- or propyl-paraben and chlorobutanol. The pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent and the like in addition to the above components. Other pharmaceutically acceptable carriers and formulations may be referred to as described in Remington's Pharmaceutical Sciences, 19th ed., Mack Publishing Company, Easton, PA, 1995.
본 발명의 조성물은 인간을 비롯한 포유동물에 어떠한 방법으로도 투여할 수 있다. 예를 들면, 경구 또는 비경구적으로 투여할 수 있다. 비경구적인 투여방법으로는 이에 한정되지는 않으나, 정맥내, 근육내, 동맥내, 골수내, 경막내, 심장내, 경피, 피하, 복강내, 비강내, 장관, 국소, 설하 또는 직장내 투여일 수 있다.The composition of the present invention may be administered to any mammal, including humans. For example, it can be administered orally or parenterally. Parenteral administration methods include, but are not limited to, intravenous, intramuscular, intraarterial, intramedullary, intradural, intracardiac, transdermal, subcutaneous, intraperitoneal, intranasal, intestinal, topical, sublingual or rectal administration. Can be.
본 발명의 약학적 조성물은 상술한 바와 같은 투여 경로에 따라 경구 투여용 또는 비경구 투여용 제제로 제형화할 수 있다.The pharmaceutical composition of the present invention may be formulated into a preparation for oral or parenteral administration according to the route of administration as described above.
경구 투여용 제제의 경우, 본 발명의 조성물은 분말, 과립, 정제, 환제, 당의정제, 캡슐제, 액제, 겔제, 시럽제, 슬러리제, 현탁액 등으로 당업계에 공지된 방법을 이용하여 제형화될 수 있다. 예를 들어, 경구용 제제는 활성성분을 고체 부형제와 배합한 다음 이를 분쇄하고 적합한 보조제를 첨가한 후 과립 혼합물로 가공함으로써 정제 또는 당의정제를 수득할 수 있다. 적합한 부형제의 예로는 락토즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨 및 말티톨 등을 포함하는 당류와 옥수수 전분, 밀 전분, 쌀 전분 및 감자 전분 등을 포함하는 전분류, 셀룰로즈, 메틸 셀룰로즈, 나트륨 카르복시메틸셀룰로오즈 및 하이드록시프로필메틸-셀룰로즈 등을 포함하는 셀룰로즈류, 젤라틴, 폴리비닐피롤리돈 등과 같은 충전제가 포함될 수 있다. 또한, 경우에 따라 가교결합 폴리비닐피롤리돈, 한천, 알긴산 또는 나트륨 알기네이트 등을 붕해제로 첨가할 수 있다. 나아가, 본 발명의 약학적 조성물은 항응집제, 윤활제, 습윤제, 향료, 유화제 및 방부제 등을 추가로 포함할 수 있다.For formulations for oral administration, the compositions of the present invention may be formulated using methods known in the art as powders, granules, tablets, pills, dragees, capsules, solutions, gels, syrups, slurries, suspensions, and the like. Can be. For example, oral formulations can be obtained by tablets or dragees by combining the active ingredients with solid excipients and then grinding them, adding suitable auxiliaries and processing them into granule mixtures. Examples of suitable excipients include sugars, including lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol and maltitol and starch, cellulose, including starch, corn starch, wheat starch, rice starch and potato starch, and the like. Fillers such as cellulose, gelatin, polyvinylpyrrolidone, and the like, including methyl cellulose, sodium carboxymethylcellulose, hydroxypropylmethyl-cellulose, and the like. In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid or sodium alginate and the like may optionally be added as a disintegrant. Furthermore, the pharmaceutical composition of the present invention may further include an anticoagulant, a lubricant, a humectant, a perfume, an emulsifier, a preservative, and the like.
비경구 투여용 제제의 경우에는 주사제, 크림제, 로션제, 외용연고제, 오일제, 보습제, 겔제, 에어로졸 및 비강 흡입제의 형태로 당업계에 공지된 방법으로 제형화할 수 있다. 이들 제형은 모든 제약 화학에 일반적으로 공지된 처방서인 문헌(Remington's Pharmaceutical Science, 19th ed., Mack Publishing Company, Easton, PA,1995)에 기재되어 있다.Formulations for parenteral administration may be formulated by methods known in the art in the form of injections, creams, lotions, external ointments, oils, humectants, gels, aerosols and nasal inhalants. These formulations are described in Remington's Pharmaceutical Science, 19th ed., Mack Publishing Company, Easton, PA, 1995, a prescription generally known in all pharmaceutical chemistries.
본 발명의 조성물의 총 유효량은 단일 투여량(single dose)으로 환자에게 투여될 수 있으며, 다중 투여량(multiple dose)으로 장기간 투여되는 분할 치료 방법(fractionated treatment protocol)에 의해 투여될 수 있다. 본 발명의 약학적 조성물은 질환의 정도에 따라 유효성분의 함량을 달리할 수 있다. 바람직하게 본 발명의 약학적 조성물의 바람직한 전체 용량은 1일당 환자 체중 1㎏당 약 0.01㎍ 내지 10,000㎎, 가장 바람직하게는 0.1㎍ 내지 1,000㎎일 수 있다. 그러나 상기 약학적 조성물의 용량은 제제화 방법, 투여 경로 및 치료 횟수뿐만 아니라 환자의 연령, 체중, 건강 상태, 성별, 질환의 중증도, 식이 및 배설율 등 다양한 요인들을 고려하여 환자에 대한 유효투여량이 결정되는 것이므로, 이러한 점을 고려할 때 당 분야의 통상적인 지식을 가진 자라면 본 발명의 조성물의 적절한 유효 투여량을 결정할 수 있을 것이다. 본 발명에 따른 약학적 조성물은 본 발명의 효과를 보이는 한 그 제형, 투여 경로 및 투여 방법에 특별히 제한되지 아니한다. The total effective amount of the composition of the present invention may be administered to a patient in a single dose and may be administered by a fractionated treatment protocol which is administered in multiple doses for a long time. The pharmaceutical composition of the present invention may vary the content of the active ingredient depending on the extent of the disease. Preferably the preferred total dose of the pharmaceutical composition of the present invention may be from about 0.01 μg to 10,000 mg, most preferably 0.1 μg to 1,000 mg per kg body weight of the patient per day. However, the dosage of the pharmaceutical composition is determined in consideration of various factors such as the formulation method, route of administration and frequency of treatment, as well as various factors such as the patient's age, weight, health status, sex, severity of the disease, diet and excretion rate. In view of this, one of ordinary skill in the art will be able to determine the appropriate effective dosage of the compositions of the present invention. The pharmaceutical composition according to the present invention is not particularly limited to its formulation, route of administration and method of administration as long as the effect of the present invention is shown.
본 발명의 다른 양태인 길초근, 당귀, 천궁 및 작약의 혼합 추출물을 유효성분으로 포함하는 갱년기 증상 개선, 치료 및 면역효능을 갖는 식품 조성물을 제공한다.It provides a food composition having menopausal symptom improvement, treatment and immune efficacy comprising a mixed extract of Gilcho root, Angelica, Cervix and Peony, which is another embodiment of the present invention, as an active ingredient.
본 발명의 상기 식품 조성물은 기능성 식품(functional food), 영양 보조제(nutritional supplement), 건강식품(health food)및 식품 첨가제(food additives)등의 모든 형태를 포함한다. 상기 유형의 식품 조성물은 당업계에 공지된 통상적인 방법에 따라 다양한 형태로 제조할 수 있다.The food composition of the present invention includes all forms such as functional food, nutritional supplement, health food and food additives. Food compositions of this type can be prepared in various forms according to conventional methods known in the art.
예를 들면, 건강식품으로는 상기 조성물 자체를 차, 주스 및 드링크의 형태로 제조하여 음용하도록 하거나, 과립화, 캡슐화 및 분말화하여 섭취할 수 있다.For example, the health food may be prepared by drinking the composition itself in the form of tea, juice and drink, or ingested by granulation, encapsulation and powdering.
또한, 기능성 식품으로는 음료(알콜성 음료 포함), 과실 및 그의 가공식품(예: 과일통조림, 병조림, 잼, 마말레이드 등), 어류, 육류, 및 그 가공식품(예: 햄, 소시지 콘비프 등), 빵류 및 면류(예: 우동, 메밀국수, 라면, 스파게티, 마카로니 등), 과즙, 각종 드링크, 쿠키, 엿, 유제품(예: 버터, 치즈 등), 식용식물유지, 마아가린, 식물성 단백질, 레토르트 식품, 냉동식품, 각종 조미료(예: 된장, 간장, 소스 등)등에 추출물을 첨가하여 제조할 수 있다. 또한, 본 발명의 조성물을 식품 첨가제의 형태로 사용하기 위해서는 분말 또는 농축액 형태로 제조하여 사용할 수 있다. Functional foods also include beverages (including alcoholic beverages), fruits and processed foods (e.g. canned fruit, canned foods, jams, marmalade, etc.), fish, meat, and processed foods (e.g. ham, sausage corned beef, etc.). , Breads and noodles (e.g. udon, soba, ramen, spaghetti, macaroni, etc.), fruit juices, various drinks, cookies, malts, dairy products (e.g. butter, cheese, etc.), edible vegetable oils, margarine, vegetable protein, retort food , Frozen foods, various seasonings (eg, miso, soy sauce, sauce, etc.) can be prepared by adding the extract. In addition, in order to use the composition of the present invention in the form of a food additive, it can be prepared in powder or concentrate form.
본 발명의 식품용 조성물 중 상기 추출물의 바람직한 함량은 식품용 조성물 총 중량에 대하여 0.001 내지 50% 일 수 있으며, 바람직하게는 0.1 내지 50% 범위로 함유될 수 있다.The preferred content of the extract in the food composition of the present invention may be 0.001 to 50% with respect to the total weight of the food composition, preferably contained in the range of 0.1 to 50%.
이하, 실시예 및 실험예를 들어 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail with reference to Examples and Experimental Examples.
실시예Example
시중에서 구입한 당귀 50g, 천궁 20g, 작약 20g 및 길초근 10g을 추출기에 넣고 30%(v/v)에탄올 10배수를 넣어 85 내지 90℃에서 3시간씩 2회 추출하고, 추출액을 1㎛ 크기의 마이크로필터를 이용하여 여과한다. 여과된 액을 60℃ 이하에서 감압농축 및 건조하여 혼합분말 형태의 약학적 조성물을 제조하였다.50g of commercial Angelica, 20g of Cheongung, 20g of Peony and 10g of Kilcho root were put into the extractor, and 10 times of 30% (v / v) ethanol was extracted twice for 3 hours at 85 to 90 ° C. It is filtered using a micro filter. The filtered solution was concentrated under reduced pressure at 60 ° C. or lower, and dried to prepare a pharmaceutical composition in the form of a mixed powder.
비교예Comparative example
시판중인 17-에스트라디올(17-estradiol, SIGMA에서 구입)을 준비하였다.Commercial 17-estradiol (17-estradiol, available from SIGMA) was prepared.
실험예Experimental Example 1 One
실시예에서 제조된 약학적 조성물(HPC 1) 및 비교예의 17-에스트라디올(E2)의 에스트로겐 활성을 측정하기 위하여 하기 방법(1 및 2 참조)으로 실험하였고 그에 대한 결과를 도 1에 나타내었다. 또한, 에스트로겐 수용체 발현을 측정하기 위해 하기 방법(1 및 3 참조)으로 실험하였고 그에 대한 결과를 도 2에 나타내었다.In order to measure the estrogen activity of the pharmaceutical composition prepared in Example (HPC 1) and the comparative example 17-estradiol (E2) was tested by the following method (see 1 and 2) and the results are shown in FIG. In addition, the following methods (see 1 and 3) were used to measure estrogen receptor expression and the results are shown in FIG. 2.
<실험방법>Experimental Method
(1) 세포배양(1) Cell culture
ER(Estrogen Receptor)을 가지고 있는 인간 유방암 세포인 MCF-7 세포를 CO2 인큐베이터(5% CO2, 37℃)에서 10% 소 태아혈청(FBS : Fetal Bovine Serum), 0.01㎎/㎖의 소 인슐린(bovine insulin) 및 항생제(100 unit/㎖ penicillin 및 100㎍/㎖ streptomycin)를 첨가한 RPMI-1640 배지로 3 내지 4일마다 배지를 교환하고, 7일마다 계대 배양을 통하여 사용하였다. MCF-7 cells, human breast cancer cells with ER (Estrogen Receptor), were cultured in a CO 2 incubator (5% CO 2 , 37 ° C) in 10% Fetal Bovine Serum (FBS) and 0.01 mg / ml bovine insulin. (bovine insulin) and antibiotics (100 unit / ml penicillin and 100 µg / ml streptomycin) were added to the medium every 3-4 days with RPMI-1640 medium and used every seven days.
ER을 가지고 있지 않은 인간 유방암 세포인 MDA-MB231 세포를 CO2 인큐베이터(5% CO2, 37℃)에서 10% 소태아혈청(FBS) 및 항생제(100 unit/㎖ penicillin 및 100㎍/㎖ streptomycin)를 첨가한 RPMI-1640 배지로 4 내지 5일마다 배지를 교환하고, 6 내지 7일마다 계대 배양을 통하여 사용하였다.MDA-MB231 cells, human breast cancer cells without ER, were treated with 10% fetal bovine serum (FBS) and antibiotics (100 unit / ml penicillin and 100 µg / ml streptomycin) in a CO 2 incubator (5% CO 2 , 37 ° C). The medium was changed every 4 to 5 days with RPMI-1640 medium added thereto, and used through subculture every 6 to 7 days.
(2) 에스트로겐 활성 측정(2) determination of estrogen activity
에스트로겐 활성에 의한 세포증식 효과의 측정은 Soto 등의 방법(Soto et al., The E-SCREEN assay as a tool to identify estrogens: anupdate on estrogenic enviromental pollutants. Envirom Health Pers, 103, 113∼22, 1995)을 일부 변형하여 적용하였다.The measurement of cell proliferation effect by estrogen activity was performed by Soto et al. (Soto et al., The E-SCREEN assay as a tool to identify estrogens: anupdate on estrogenic enviromental pollutants. Envirom Health Pers, 103, 113-22, 1995). Some variations were applied.
MCF-7 세포 및 MDA-MB 231 세포는 배지 중의 에스트로겐 활성을 나타내는 성분을 배제하기 위하여, 10% 숯으로 처리된 소 태아혈청(charcoal treated FBS)이 함유된 페놀레드(phenol red)를 포함하지 않는 RPMI-1640 배지로 96 well plate에 well당 2Х103개를 seeding하고, 24시간 후 test medium(5% charcoal dextran treated FBS를 함유한 페놀레드를 포함하지 않는 RPMI 1640 배지)으로 교환하였다. 이후, 실시예에서 제조된 조성물(HPC 1)을 증류수로 용해하여 10, 25, 50, 100 및 200㎍/㎖를 첨가하였고, 비교예의 1nM인 17-에스트라디올(E2)을 첨가하였다. 또한, 대조군으로 증류수를 첨가하였다. 첨가 후, well당 MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)염료를 첨가하여 에스트로겐 활성을 측정하였다. 그에 따른 결과를 도 1에 나타내었다.MCF-7 cells and MDA-
(3) 에스트로겐 수용체 발현 측정 (3) estrogen receptor expression measurement
에스트로겐 활성을 확인하기 위한 에스트로겐 수용체에 대한 실험(reporter assay)은 인디고 바이오 사이언스사에서 구입한 human ER reporter assay kit를 사용하여 4시간 동안 수행되었다.Reporter assays for estrogen receptors to confirm estrogen activity were performed for 4 hours using a human ER reporter assay kit purchased from Indigo Bioscience.
실시예 1에서 제조된 약학적 조성물(HPC 1) 및 비교예의 1nM인 17-에스트라디올(E2)를 증류수에 용해한 후, human ER이 코팅된 well에 첨가하여 4시간 후, 세척한 후 assay solution을 각 well에 첨가하여 결합능를 측정하였고, 대조군으로 증류수를 이용하여 동일한 방법으로 결합능을 측정하였다.The pharmaceutical composition (HPC 1) prepared in Example 1 and 17-estradiol (E2), which is 1 nM of Comparative Example, were dissolved in distilled water, and then added to human ER-coated wells. The binding capacity was measured by adding to each well, and the binding capacity was measured by the same method using distilled water as a control.
상기 에스트라디올은 에스트로겐 수용체의 리간드로 작용하는데, 특히 ER α(Estrogen Receptor α) 및 ER β(Estrogen Receptor β)에 작용한다.The estradiol acts as a ligand of the estrogen receptor, and in particular, acts on ER α (Estrogen Receptor α) and ER β (Estrogen Receptor β).
UN : 증류수를 투여한 대조군UN: control group administered distilled water
HPC 1 : 실시예에서 제조된 조성물을 투여한 처리군HPC 1: Treatment group to which the composition prepared in Example was administered
E2 : 비교예의 17-에스트라디올을 투여한 처리군E2: Treatment group in which 17-estradiol of Comparative Example was administered
도 1은 실시예 및 비교예에 따른 물질 투여 후 에스트로겐 활성을 측정하여 나타낸 그래프이고, 도 2는 실시예 및 비교예에 따른 물질 투여 후 에스트로겐 수용체 발현을 측정하여 나타낸 그래프이다. 1 is a graph showing the measurement of estrogen activity after administration of a substance according to Examples and Comparative Examples, Figure 2 is a graph showing the measurement of estrogen receptor expression after administration of a substance according to Examples and Comparative Examples.
시료의 에스트로겐 활성 확인은 아무런 처치도 하지 않은 대조군의 세포 생존율을 100%로 환산하여 판단하였다. 도 1의 (a)를 참조하면 에스트로겐 수용체(ER)가 내재적으로 발현되어 있는 MCF-7 세포에서 17-에스트라디올(비교예, E2)을 첨가한 경우, 세포 생존율이 118.3 ± 8.72%로 대조군 대비 세포 생존율이 유의적으로 증가되었고, 약학적 조성물(실시예, HPC 1)을 첨가한 경우는 25㎍/㎖에서 대조군 대비 세포 생존율이 10.0 %(110.0± 5.03) 유의적으로 증가되었다. Confirmation of the estrogen activity of the sample was determined by converting the cell viability of the control group without any treatment to 100%. Referring to (a) of FIG. 1, when 17-estradiol (Comparative Example, E2) is added to MCF-7 cells in which the estrogen receptor (ER) is intrinsically expressed, the cell survival rate is 118.3 ± 8.72% compared to the control group. Cell viability was significantly increased, and when the pharmaceutical composition (Example, HPC 1) was added, the cell viability was 10.0% (110.0 ± 5.03) compared to the control at 25 µg / ml. Significantly increased.
또한, 도 1의 (b)를 참조하면 에스트로겐 수용체가 발현되지 않은 MDA-MB 231에서 17-에스트라디올(비교예, E2)을 첨가한 경우는 세포 생존율이 109.4 ± 7.56%로 대조군 대비 세포 생존율이 유의적으로 증가되었고, 약학적 조성물(실시예, HPC 1)을 첨가한 경우는 250㎍/㎖에서 대조군 대비 세포 생존율이 6.0%(106.0± 5.21)유의적으로 증가되었다.In addition, referring to FIG. 1 (b), when 17-estradiol (Comparative Example, E2) was added in MDA-
도 2에서 (a)를 참조하면 17-에스트라디올(비교예, E2)은 루시페라아제의 활성도가 2.36 ± 0.49%로 대조군 대비 유의적으로 증가하였고, 약학적 조성물(실시예, HPC 1)은 50㎍/㎖ 처리군에서 루시페라아제의 활성도가 대조군 대비 1.8% (1.82± 0.18) 유의적으로 증가되었다. Referring to (a) of FIG. 2, 17-estradiol (Comparative Example, E2) significantly increased luciferase activity to 2.36 ± 0.49% compared to the control group, and the pharmaceutical composition (Example, HPC 1) was 50 μg. Luciferase activity was significantly increased by 1.8% (1.82 ± 0.18) compared to the control group.
또한, (b)를 참조하면, 17-에스트라디올(비교예, E2)은 루시페라아제의 활성도가 2.93± 0.12%로 대조군 대비 유의적으로 증가하였고, 약학적 조성물(실시예, HPC 1)은 100㎍/㎖ 처리군에서 루시페라아제의 활성도가 대조군 대비 2.7% (2.73± 0.02) 유의적으로 증가되어, 에스트로겐과 유사한 활성을 나타내었다.In addition, referring to (b), 17-estradiol (Comparative Example, E2) significantly increased the activity of luciferase to 2.93 ± 0.12% compared to the control, pharmaceutical composition (Example, HPC 1) was 100㎍ Luciferase activity was significantly increased by 2.7% (2.73 ± 0.02) compared to the control group in the / ml treatment group, showing an estrogen-like activity.
실험예Experimental Example 2 2
실시예에서 제조된 약학적 조성물(HPC 1), 비교예의 17-에스트라디올(E2) 투여에 따른 알칼리성 인산분해효소(ALP)의 변화, 혈중 에스트로겐의 변화, 황체형성호르몬(LH)의 변화, 여포자극호르몬(FSH)의 변화, 자궁 무게 및 자궁 두께 변화를 관찰하기 위해 하기와 같은 방법으로 실험하였고 그에 대한 결과를 각각 도 3 내지 6에 나타내었다.Pharmaceutical composition prepared in Example (HPC 1), changes in alkaline phosphatase (ALP), blood estrogen, changes in luteinizing hormone (LH), follicles following administration of 17-estradiol (E2) of Comparative Example In order to observe the change in stimulating hormone (FSH), uterine weight and uterine thickness change, the experiment was performed as follows, and the results thereof are shown in FIGS. 3 to 6, respectively.
<실험방법>Experimental Method
(1) 난소 절제 시술 및 동물 사육(1) Ovariectomy and Animal Breeding
실험동물을 케타민(Keara 100㎎/㎏) 및 2%의 자일라진(Rumpun 0.15㎖/㎏)으로 전신 마취 후, 통법에 따라 제모 및 무균 처리(10% povine-iodine scrub followed by 70% alcohol wipe)하였다. 이후, 난소 절제 그룹(OVX)은 실험동물의 등쪽 중앙을 1cm가량의 절개한 후 봉합용 실로 난소를 결찰하고, 양측의 난소를 절제하였다. 난소 절제 후 각 장기를 복강 내로 재위치 시킨 후 봉합용 실로 층별 봉합하였다. 반면, 난소 비(非)절제 그룹(SHAM)은 난소는 적출하지 않고 난소 절제와 같은 스트레스를 주기 위하여 모의수술(Sham operation)을 시행하였고, 1주일의 회복 과정을 거쳤다. Animals were subjected to general anesthesia with ketamine (
난소 절제 후 난소 절제 그룹(OVX)에 실시예에서 제조된 조성물(HPC 1)을 200㎎/㎏ 및 비교예의 1nm인 에스트라디올(E2)은 10㎍/㎏를 매일 같은 시간에 생리 식염수와 함께 3개월간 경구 투여하였고, 대조군은 동일 양의 생리 식염수를 3개월 동안 경구 투여하였다. 사육기간 중에는 미국영양학회의 실험쥐를 위한 표준사료인 AIN-76사료와 증류수를 자유롭게 섭취하도록 하였다. After ovarian resection, the ovarian resection group (OVX) used 200 mg / kg of the composition (HPC 1) prepared in Example and estradiol (E2), which is 1 nm of the comparative example, at 10 占 // kg with physiological saline at the same time every day. The group was orally administered, and the control group was orally administered the same amount of saline solution for 3 months. During the breeding period, AIN-76 feed and distilled water were ingested freely.
(2) ALP 변화량 측정 (2) ALP variation measurement
모든 시험 종료 후 수득한 혈청을 활용하여 ALP의 유도 정도를 혈액생화학장비(7170S, Hitachi, Tokyo, Japan)를 활용 하여 측정하고, 그에 따른 결과를 도 3에 나타내었다.The induction of ALP was measured using blood biochemical equipment (7170S, Hitachi, Tokyo, Japan) using the serum obtained after the completion of all the tests, and the results are shown in FIG. 3.
(3) 에스트라디올 측정(3) Estradiol measurement
4주 간격으로 안와정맥채혈을 수행하여 혈청을 수득하였다. 시험 종료 시, 실험동물을 에테르 마취 후 개복하여 복대정맥에서 혈액을 채취하였다. 채취 후 혈액을 헤파린 처리된 튜브로 옮겨 담고 2,500rpm(4℃)에서 15분간 원심분리하여 혈청을 분리하였다. 분리된 혈청을 대상으로 Testosterone, Estradiol ELISA kit(ADI-065, 174; Enzo Life Sciences, U.S.A.)를 이용하여 혈중 에스트라디올 함량을 측정하고, 그에 따른 결과를 도 4에 나타내었다.Orbital venous blood collection was performed at 4 week intervals to obtain serum. At the end of the test, the experimental animals were opened after ether anesthesia and blood was collected from the abdominal vein. After collection, the blood was transferred to a heparinized tube, and serum was separated by centrifugation at 2,500 rpm (4 ° C.) for 15 minutes. Blood estradiol content was measured using the Testosterone, Estradiol ELISA kit (ADI-065, Enzo Life Sciences, U.S.A.) in the isolated serum, and the results are shown in FIG. 4.
(4)LH/FSH 측정 (4) LH / FSH measurement
모든 시험 종료 후 분리된 혈청을 대상으로 황체형성호르몬(LH) 및 여포자극호르몬(FSH) Elisa kit(ADI-065, 174; Enzo Life Sciences, U.S.A.)를 이용하여 각 변화량을 측정하였고, 그에 따른 결과를 도 5에 나타내었다.After the completion of all the tests, the serum was isolated from the luteinizing hormone (LH) and follicle stimulating hormone (FSH) Elisa kit (ADI-065, ® Enzo Life Sciences, USA) to measure the amount of change. Is shown in FIG. 5.
(5) 조직학적 검사 (5) histological examination
동물실험 종료 후, 얻은 난소 조직을 필터페이퍼에 편평하게 부착하여 4% 포르말린 용액에 4시간 동안 고정 시킨 후, 조직 처리 과정을 거쳐 파라핀 포매하였다. 포매 후 조직 절편기를 이용하여 난소 조직을 5㎛두께로 절단하여 슬라이드에 붙이고, 조직 절편은 자일렌을 이용하여 파라핀을 제거하고 알코올과 증류수로 함수 시켰다. 함수 후 피부 조직 관찰을 위해 H&E(Hematoxylin & Eosin)염색하고 현미경으로 관찰한 결과를 도 6에 나타내었다.After the end of the animal experiment, the obtained ovarian tissue was flatly attached to the filter paper and fixed in 4% formalin solution for 4 hours, and then paraffin embedded through the tissue treatment process. After embedding, the tissue section was cut into ovarian tissue with a thickness of 5 μm and attached to the slide. The tissue section was paraffin-free with xylene and hydrated with alcohol and distilled water. 6 shows the results of staining with H & E (Hematoxylin & Eosin) for observing skin tissue after hydration and microscopic observation.
OVX : 난소 절제 그룹.OVX: Ovarian Ablation Group.
SHAM : 난소 비 절제 그룹.SHAM: Ovarian non-ablation group.
HPC 1 또는 OVX+HPC 1 : 난소 절제 그룹에 실시예에서 제조된 약학적 조성물을 투여한 처리군.
E2 또는 OVX+E2 : 난소 절제 그룹에 비교예의 17-에스트라디올을 투여한 처리군.E2 or OVX + E2: Treatment group in which 17-estradiol of Comparative Example was administered to ovarian ablation group.
도 3은 실시예 및 비교예에 따른 물질 투여한 후 ALP 변화량을 측정한 그래프이고, 도 4는 실시예 및 비교예에 따른 물질 투여 후 혈중 에스트로겐 농도를 측정한 그래프이고, 도 5는 실시예 및 비교예에 따른 물질 투여 후 혈중 LH 및 FSH의 변화를 나타낸 그래프이고, 도 6은 실시예 및 비교예에 따른 물질 투여 후 자궁무게를 나타낸 그래프 및 자궁두께를 현미경으로 관찰한 사진이다.Figure 3 is a graph measuring the amount of ALP change after administration of the substance according to the Examples and Comparative Examples, Figure 4 is a graph measuring the concentration of estrogen in the blood after administration of the substance according to the Examples and Comparative Examples, Figure 5 A graph showing changes in blood LH and FSH after administration of a substance according to a comparative example, and FIG. 6 is a graph showing a graph showing uterine weight and uterine thickness after administration of a substance according to Examples and Comparative Examples under a microscope.
도 3을 참조하면, 난소 절제 1주차부터 SHAM 그룹 대비 모든 그룹에서 혈중ALP량이 유의적으로 증가한 것을 알 수 있다. 17-에스트라디올 10㎍/㎏ 투여한 경우(비교예, E2)에는 1달 후부터 SHAM과 비슷한 수준의 혈중 ALP량을 나타내었다. 투여 3개월 후 SHAM그룹(46.5±4.0U/L) 대비 난소 절제 그룹(89.0±4.0 U/L)의 혈중 ALP량은 47.7% 증가되었다. Referring to FIG. 3, it can be seen that the blood ALP amount was significantly increased in all groups compared to the SHAM group from the first week of ovarian resection. When 10 μg / kg of 17-estradiol was administered (Comparative Example, E2), the blood ALP level was similar to that of SHAM after one month. Three months after the administration, the ALP level in the ovarian ablation group (89.0 ± 4.0 U / L) was increased by 47.7% compared to the SHAM group (46.5 ± 4.0 U / L).
반면, 17-에스트라디올 투여 그룹(비교예, E2)의 혈중 ALP량은 50.0±1.0 U/L로 난소 절제 그룹 대비 43.7% 감소되었고, 당귀, 천궁, 작약 및 길초근을 일정비율로 첨가하여 제조된 약학적 조성물(실시예, HPC 1) 투여 그룹의 혈중 ALP량은 68.03±4.03 U/L로 난소 절제 그룹 대비 23.6% 유의적 감소되었다. On the other hand, the blood ALP level of the 17-estradiol group (Comparative Example, E2) was 50.0 ± 1.0 U / L, which was reduced by 43.7% compared to the ovarian ablation group. The amount of ALP in the blood of the administered pharmaceutical composition (Example, HPC 1) group was 68.03 ± 4.03 U / L, which was 23.6% lower than that of the ovarian ablation group.
도 4를 참조하면, 난소 절제 1주차부터 난소절제 그룹 대비 SHAM 그룹의 혈중 에스트로겐의 농도는 유의적으로 증가되었다. 17-에스트라디올을 투여(비교예, E2)한 그룹은 1달 후부터 SHAM과 비슷한 수준의 혈중 에스트로겐의 농도를 나타내었다. 투여 3개월 후 SHAM그룹(62.8±4.03 pg/㎖) 대비 난소 절제 그룹(32.0±2.03 pg/㎖)의 혈중 에스트로겐량의 농도는 96.2% 감소되었다. Referring to FIG. 4, the blood estrogen concentration of the SHAM group was significantly increased from the ovarian resection group from the first week. Groups treated with 17-estradiol (Comparative Example, E2) showed levels of blood estrogen similar to SHAM from 1 month later. Three months after administration, the concentration of blood estrogen in the ovarian ablation group (32.0 ± 2.03 pg / ml) was reduced by 96.2% compared to the SHAM group (62.8 ± 4.03 pg / ml).
반면, 17-에스트라디올 투여 그룹(비교예, E2)의 혈중 에스트로겐의 농도는 70.0±4.03pg/㎖로 난소 절제 그룹 대비 118.6% 증가되었고, 당귀, 천궁, 작약 및 길초근을 일정비율로 첨가하여 제조된 약학적 조성물(실시예, HPC 1) 투여 그룹의 혈중 에스트로겐의 농도는 41.5±2.4pg/㎖로 난소 절제 그룹 대비 10.3% 증가되었다.On the other hand, the concentration of estrogen in the 17-estradiol group (Comparative Example, E2) was 70.0 ± 4.03 pg / ml, which was 118.6% higher than that in the ovarian ablation group. The concentration of blood estrogen in the prepared pharmaceutical composition (Example, HPC 1) administration group was 41.5 ± 2.4 pg / ml, which was increased by 10.3% compared to the ovarian ablation group.
도 5의 (a)를 참조하면, 난포자극 호르몬 측정 결과, SHAM그룹(210±4.6 ng/㎖) 대비 난소 절제 그룹의 혈중 난포자극 호르몬 농도는 295.4±10.4ng/㎖로 28.8% 증가되었다. Referring to (a) of FIG. 5, as a result of measuring the follicle stimulating hormone, the concentration of the follicle stimulating hormone in the ovarian ablation group compared to the SHAM group (210 ± 4.6 ng / ml) was increased by 28.8% to 295.4 ± 10.4ng / ml.
반면, 17-에스트라디올 투여 그룹(비교예, E2)의 혈중 난포자극 호르몬 농도는 245.2±6.2 ng/㎖로 난소 절제 그룹 대비 16.9% 감소되었고, 당귀, 천궁, 작약 및 길초근을 일정비율로 첨가하여 제조된 약학적 조성물(실시예, HPC 1) 투여 그룹의 혈중 난포자극 호르몬 농도는 275.4±5.0 ng/㎖로 난소 절제 그룹 대비 6.8% 유의적으로 감소하였다.On the other hand, the concentration of follicle stimulating hormone in the 17-estradiol group (Comparative Example, E2) was 245.2 ± 6.2 ng / ml, which was reduced by 16.9% compared to the ovarian ablation group. The concentration of follicle stimulating hormone in the pharmaceutical composition (Example, HPC 1) administered group was 275.4 ± 5.0 ng / ml, which was significantly decreased by 6.8% compared to the ovarian ablation group.
또한, (b)를 참조하면, 황체 자극 호르몬 측정 결과, SHAM그룹 (62.4±2.1 ng/㎖) 대비 난소 절제 그룹의 혈중 황체 자극 호르몬 농도는 114.0±2.0 ng/㎖로 45.2% 증가되었다. In addition, referring to (b), as a result of measuring the progesterone hormone, the concentration of progesterone hormone in the blood of the ovarian resection group compared to the SHAM group (62.4 ± 2.1 ng / ml) was increased by 45.2% to 114.0 ± 2.0 ng / ml.
반면, 17-에스트라디올 투여 그룹(비교예, E2)의 혈중 황체 자극 호르몬 농도는 68.3±5.2 ng/㎖로 난소 절제 그룹 대비 40.1% 감소되었고, 당귀, 천궁, 작약 및 길초근을 일정비율로 첨가하여 제조된 약학적 조성물(실시예, HPC 1) 투여 그룹의 혈중 황체 자극 호르몬 농도는 98.2±3.4 ng/㎖로 난소 절제 그룹 대비 10.1% 감소되었다.In contrast, the concentration of progesterone hormone in the 17-estradiol group (Comparative Example, E2) was 68.3 ± 5.2 ng / ml, which was 40.1% lower than that of the ovarian ablation group. The concentration of progesterone hormone in the blood of the pharmaceutical composition (Example, HPC 1) administered group was 98.2 ± 3.4 ng / ml, which was reduced by 10.1% compared to the ovarian ablation group.
도 6을 참조하면, 난소 절제 그룹의 자궁무게는 SHAM 그룹 대비 자궁무게가 7.4배 감소하였다. 반면, 17-에스트라디올 투여 그룹(비교예, E2)의 자궁무게는 난소 절제 그룹 대비 6.3배 증가되었다Referring to FIG. 6, the uterine weight of the ovarian ablation group was 7.4 times lower than that of the SHAM group. In contrast, the uterine weight of the 17-estradiol group (Comparative Example, E2) was 6.3 times higher than that of the ovarian ablation group.
반면, 당귀, 천궁, 작약 및 길초근을 일정비율로 첨가하여 제조된 약학적 조성물(실시예, HPC 1) 투여 그룹의 자궁무게는 난소 절제 그룹과 비교하여 0.67배 증가 되었으나 유의적 차이는 관찰되지 않았다. 다만, 자궁에서 호르몬에 영향을 받는 부분은 자궁속막 기능층으로 에스트로겐의 영향이 없는 난소 절제 그룹에서는 자궁의 두께가 얇아졌으나, 약학적 조성물(실시예, HPC 1) 투여 그룹의 자궁속막 기능층 두께는 두꺼워진 것을 확인할 수 있었다.On the other hand, the weight of the uterine in the pharmaceutical composition (Example, HPC 1) administered with the addition of Angelica, Cervix, Peony and Gilcho root was increased by 0.67 times compared with the Ovarian excision group, but no significant difference was observed. Did. However, the uterine thickness in the uterine ablation group without the effect of estrogen is thinner in the uterine membrane. However, the thickness of the uterine endometrium in the pharmaceutical composition (Example, HPC 1) administration group Could be thickened.
상기 실험예 1 내지 2을 참조하면 본 발명인 길초근, 당귀, 천궁 및 작약의 혼합 추출물을 유효성분으로 포함하는 약학적 조성물은 에스트로겐과 비슷한 효능이 있으므로, 갱년기 증상에 효능이 있음을 알 수 있다.Referring to Experimental Examples 1 to 2, it can be seen that the pharmaceutical composition comprising a mixed extract of the present invention Gilchogeun, Angelica, Cnidium and Peony as an active ingredient has an effect similar to estrogen, and thus is effective in menopausal symptoms.
또한, 길초근, 당귀, 천궁 및 작약의 혼합 추출물을 유효성분으로 포함하는 식품조성물 역시 갱년기 증상에 효능이 있을 것으로 기대된다.In addition, it is expected that the food composition containing a mixed extract of Gilcho root, Angelica, cheonggung and peony as an active ingredient is also effective in menopausal symptoms.
이상으로 본 발명의 바람직한 실시예를 도면을 참고하여 상세하게 설명하였다. 본 발명의 설명은 예시를 위한 것이며, 본 발명이 속하는 기술분야에서 통상의 지식을 가진 자는 본 발명의 기술적 사상이나 필수적인 특징을 변경하지 않고서 다른 구체적인 형태로 쉽게 변형이 가능하다는 것을 이해할 수 있을 것이다.The preferred embodiment of the present invention has been described in detail above with reference to the drawings. The description of the present invention is for illustrative purposes, and it will be understood by those skilled in the art that the present invention may be easily modified in other specific forms without changing the technical spirit or essential features of the present invention.
따라서, 본 발명의 범위는 상기 상세한 설명보다는 후술하는 특허청구범위에 의하여 나타내어지며, 특허청구범위의 의미, 범위 및 그 균등 개념으로부터 도출되는 모든 변경 또는 변형된 형태가 본 발명의 범위에 포함되는 것으로 해석되어야 한다.Therefore, the scope of the present invention is shown by the claims below rather than the detailed description, and all changes or modifications derived from the meaning, scope, and equivalent concepts of the claims are included in the scope of the present invention. Should be interpreted.
Claims (6)
상기 약학적 조성물은 길초근 3 내지 18 중량%, 당귀 40 내지 60중량%, 천궁 10 내지 30중량%, 작약 10 내지 30중량%인 것을 특징으로 하는 약학적 조성물.The method of claim 1,
The pharmaceutical composition is 3 to 18% by weight of the herbaceous root, Angelica 40 to 60% by weight, 10 to 30% by weight cheongung, peony 10 to 30% by weight of the pharmaceutical composition.
상기 혼합 추출물은 물, 탄소수 1 내지 6인 알코올, 헥산, 에틸아세테이트 및 이들의 혼합용매로 이루어진 군에서 선택된 1종 이상의 용매로 추출한 것임을 특징으로 하는 조성물. The method of claim 1,
The mixed extract is a composition, characterized in that extracted with one or more solvents selected from the group consisting of water, alcohol having 1 to 6 carbon atoms, hexane, ethyl acetate and a mixed solvent thereof.
상기 알코올은 에탄올인 것을 특징으로 하는 약학적 조성물.The method of claim 3,
The alcohol is a pharmaceutical composition, characterized in that ethanol.
상기 식품 조성물은 길초근 3 내지 18 중량%, 당귀 40 내지 60중량%, 천궁 10 내지 30중량%, 작약 10 내지 30중량%인 것을 특징으로 하는 식품 조성물.
The method of claim 5,
The food composition is a food composition, characterized in that 3 to 18% by weight of Geuncho, 40 to 60% by weight, 10 to 30% by weight, 10 to 30% by weight peony.
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KR20030033473A (en) * | 2001-10-23 | 2003-05-01 | 주식회사 바이오딕스 | Compositions comprising medicinal herb extract for preventing and improving women's diseases |
KR20040010176A (en) * | 2002-07-15 | 2004-01-31 | 주식회사 한국인삼공사 | A functional food Containing herbes composition for female menopausal disorder |
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KR101855821B1 (en) * | 2017-02-28 | 2018-05-10 | 주식회사 진수바이오텍 | Composition for relieving of menopausal symptoms and improving blood circulation |
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KR20030033473A (en) * | 2001-10-23 | 2003-05-01 | 주식회사 바이오딕스 | Compositions comprising medicinal herb extract for preventing and improving women's diseases |
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