KR20070105679A - A composition for improving memory - Google Patents
A composition for improving memory Download PDFInfo
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- KR20070105679A KR20070105679A KR1020060038179A KR20060038179A KR20070105679A KR 20070105679 A KR20070105679 A KR 20070105679A KR 1020060038179 A KR1020060038179 A KR 1020060038179A KR 20060038179 A KR20060038179 A KR 20060038179A KR 20070105679 A KR20070105679 A KR 20070105679A
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Abstract
Description
도 1은 실시예 2-1의 Y-maze 실험 결과를 나타내는 그래프이다.1 is a graph showing the results of the Y-maze experiment of Example 2-1.
도 2는 실시예 2-2의 새로운 물체 인식 기억 동물 실험의 결과를 나타내는 그래프이다.Fig. 2 is a graph showing the results of the new object recognition memory animal experiment of Example 2-2.
도 3은 실시예 2-3의 수동 회피 동물 실험의 결과를 나타내는 그래프이다.3 is a graph showing the results of a passive avoidance animal experiment of Example 2-3.
도 4는 실시예 2-4의 수중 미로 동물 실험의 결과를 나타내는 그래프이다.4 is a graph showing the results of the underwater maze animal experiment of Example 2-4.
도 5는 실시예 2-5의 신경 세포 손상 방어 효과 동물 실험의 결과 사진이다. Figure 5 is a photograph of the results of the neuronal cell damage defense effect animal experiment of Example 2-5.
도 6은 실시예 3의 뇌파 측정을 통한 임상 실험에서 성인을 대상으로 한 실험 결과를 나타내는 그래프이다. Figure 6 is a graph showing the results of the experiment in adults in the clinical experiment by measuring the EEG of Example 3.
도 7은 실시예 3의 뇌파 측정을 통한 임상 실험에서 학생을 대상으로 한 실험 결과를 나타내는 그래프이다. FIG. 7 is a graph showing experimental results of students in a clinical trial using the EEG measurement of Example 3. FIG.
사람의 세포는 분화, 성장 및 파괴를 되풀이하고, 자연환경이나 음식, 운동 및 훈련과정을 통하여 세포성장은 촉진될 수 있으며, 세포 파괴를 지연시키는 물질도 우리의 생활 주변에 무수히 존재한다. 이들을 선별하여 섭취하고 활용할 때에는 장수와 함께 건강한 뇌기능을 유지할 수 있으나, 그렇지 못할 때에는 질병을 일으키거나 건망증과 치매까지 유발시키게 된다. 사람에게서 가장 중심이 되는 것은 뇌기능이라 해도 과언이 아니다. 뇌세포의 활성도가 왕성할 때 정신력이 높아지며 의지 및 결단력이 강해지고 얼굴과 전신에 기력이 충만하게 된다. 어린이에게서는 뇌세포의 활성도가 왕성할 때 뇌세포의 분화는 물론이고 성장과 판단력이 증강되며 학업성적이 향상된다. 성인은 기억력 감퇴로 인하여 매사에 잘 잊어버리며 때로 머리가 힁하고 어지러우며 곧 넘어지지 않으면 정신을 잃을 것 같은 증상을 나타내기도 한다. 이것은 뇌세포의 파괴나 신경전달물질의 분비의 부족에서도 나타나는 현상들이다. Human cells are differentiated, grown and destroyed, and cell growth can be promoted through the natural environment, food, exercise and training processes, and there are many substances that delay cell destruction around our lives. When they are screened and consumed, they can maintain healthy brain function with longevity, but otherwise they can cause disease or forgetfulness and dementia. It is no exaggeration to say that brain function is the center of man. When the activity of the brain cells is vigorous, the mental strength is increased, the will and determination will be strong, and the face and the whole body will be full of energy. In children, when the activity of brain cells is strong, not only the differentiation of brain cells, but also growth and judgment are enhanced, and academic performance is improved. Adults often forget about everything due to memory loss, and sometimes have a headache, dizziness, and loss of mind if they don't fall down. This is also seen in the destruction of brain cells and the lack of secretion of neurotransmitters.
뇌세포와 관련된 질환 중 가장 심각한 것은 치매(dementia)이다. 치매는 뇌신경의 일시적 혹은 지속적인 손상에 의한 정신기능의 전반적인 장애를 특징으로 하는 퇴행성 질환으로서 국내 사망원인 중 4위를 차지한다. 치매는 여러 가지 질환들에 의해 나타나는 병적 증상이다. 치매의 주요 원인 질환으로는 알쯔하이머병 (alzheimer disease, 이하, AD), 혈관성 치매 (Vascular dementia), 파킨슨병 (Parkinson's disease), 루이 소체 치매 (Diffuse Lewy body dementia), 헌팅톤병 (Huntington's disease), 크루츠펠트-제이야콥병 (Creutzfeldt-Jakob disease) 및 픽병 (Pick's disease) 등이 있다. 그 중, 뇌 속에 베타아밀로이드라는 독성 단백질이 축적되어 발생하는 알쯔하이머형(노인성) 치매와 뇌중풍 등으로 발생하는 뇌혈관성 치매가 전체 치매 환자의 80-90%를 차지한다. AD, 혈관성 치매, 미만성 루이소체 치매들은 치매의 증상으로만 나타날 수 있다. 치매의 증상들은 원인 질환의 종류와 정도에 따라 매우 다양하게 나타나는데, 아주 가벼운 기억장애부터 매우 심한 행동장애까지 나타나게 된다. 그러나 모든 치매 환자들은 기억장애 외에도, 사고력, 추리력 및 언어능력 등의 영역에서 어느 정도의 장애를 함께 보인다. The most serious disease associated with brain cells is dementia. Dementia is a degenerative disease characterized by overall impairment of mental function due to temporary or persistent damage to the cranial nerve, which is the fourth leading cause of death in Korea. Dementia is a pathological condition caused by various diseases. The main causes of dementia include Alzheimer's disease (AD), Vascular dementia, Parkinson's disease, Diffuse Lewy body dementia, Huntington's disease, and Crewe Creutzfeldt-Jakob disease and Pick's disease. Among them, 80-90% of all dementia patients are Alzheimer's (senile) dementia caused by accumulation of toxic protein called beta amyloid in the brain and cerebral vascular dementia caused by stroke. AD, vascular dementia and diffuse Lewy body dementia can only appear as symptoms of dementia. Symptoms of dementia vary widely depending on the type and extent of the disease, ranging from very mild memory disorders to very severe behavioral disorders. However, in addition to memory impairment, all dementia patients show some degree of impairment in areas of thinking, reasoning, and speech.
치매의 주요 발병 원인 중 하나인 AD는 주로 노인을 대상으로 심하게 나타나고 대략 50-60%가 치매로 발전한다. AD은 인지 능력이 지속적으로 감소하는 질병으로 분류되며 그 병리생리학은 복잡하나, 몇 가지의 상이한 생화학적 경로로 발병된다. 이중에 베타-아밀로이드 단백질대사, 글루타메이트성(glutamatergic), 아드레날린성(adrenergic), 세로토닌성(serotonergic), 도파민성(dopaminergic) 뉴런의 신경 전달의 이상이 그 원인으로 거론되고 있다. 그 외에도 염증, 산화, 호르몬 경로도 그 원인으로 밝혀졌다. AD 치료의 궁극적인 목표는 병 자체를 되돌려서 완치시키고 치매에 의하여 나타나는 인지장애, 정신장애, 이상행동증 등을 줄이고 없애는 것이다. 현재 많은 약들이 AD의 치료에 사용되는 것처럼 알려져 있지만 대부분의 약제들은 아직 약효에 대한 심사과정에 있다. 더욱이 현재까지 개발되고 있는 약제들을 포함해서 지금까지의 모든 약제들은 AD의 진행을 약간 늦출 수 있거나 AD에 의해 나타나는 증상에 대한 치료를 위하여 만들어진 것이지, 그 어느 것도 AD의 근본적인 병 자체를 치료할 수 있도록 고안되고 만들어진 약은 없다. 즉 AD의 발병 위험률이 높은 사람이 어떤 약물을 복용하여 병의 발병이 예방되거나 늦출 수 있다고 확실하게 인정받은 경우는 없다. 구체적으로, 상기 살펴본 바와 같이 AD의 원인이 되는 모든 경로가 치료 약물 개발의 타겟이 될 수 있지만, 최근에는 주로 아세틸콜린에스테라제(AChE) 저해제 연구가 주류를 이루고 있다. 즉, 콜린 성(cholinergic) 시냅스에서 아세틸콜린의 이용도를 증가시키는 것이다. 현재까지 4종의 AD 치료제가 FDA 승인으로 판매되고 있다: 타크린(tacrine, Cognex®), 도네페질(donepezil, Aricept®), 리바스티그민(rivastigmine, Exelon®), 갈란타민(galantamine, Reminyl®). 상기 열거한 콜린성 뉴런에 작용하는 약물들은 그 치료 효과가 너무 약하므로 좀더 강한 치료효과를 내는 비콜린신경계 약제의 개발이 시급한 실정이다. 이에 부응하여 최근에 유럽과 미국 FDA에서 중증 내지 심한 AD환자용 치료제 메만틴(memantine, Ebixa®)이 비경쟁성 NMDA 수용체 길항제로서 승인되었다.AD, one of the leading causes of dementia, is severely prevalent in older people, with approximately 50-60% of people developing dementia. AD is classified as a disease in which cognitive ability continues to decrease and its pathophysiology is complex, but develops in several different biochemical pathways. Among them, beta-amyloid protein metabolism, glutamatergic, adrenergic, serotonergic, and dopaminergic neuron abnormalities in neurotransmission have been mentioned as the cause. In addition, inflammation, oxidation and hormonal pathways have been found to be the cause. The ultimate goal of AD treatment is to reverse the disease itself, to cure it, and to reduce and eliminate the cognitive, mental, and aberrant behaviors caused by dementia. Although many drugs are now known to be used to treat AD, most drugs are still being tested for drug efficacy. Moreover, all drugs to date, including those currently being developed, are designed to slow the progression of AD slightly or to treat the symptoms caused by AD, and none of them can be used to treat the underlying disease itself. No medicine is made. In other words, a person who is at high risk of developing AD has not been clearly admitted to take any medication to prevent or slow down the onset of the disease. Specifically, as described above, all pathways that cause AD may be targets for the development of therapeutic drugs, but in recent years, research mainly on acetylcholinesterase (AChE) inhibitors has been mainstream. In other words, it increases the availability of acetylcholine at cholinergic synapses. To date, four AD therapies have been sold with FDA approval: tacrine (Cognex ® ), donepezil (Aricept ® ), rivastigmine (Exelon ® ), galantamine (Reminyl ®) ). Drugs acting on the cholinergic neurons listed above are so weak that the therapeutic effect of the non-cholinergic system that has a stronger therapeutic effect is urgently needed. In response, recently, the European and US FDA approved memantine (Ebixa ® ) for severe to severe AD patients as an uncompetitive NMDA receptor antagonist.
현재까지 국내에는 많은 기억력을 향상시키는 효능을 갖는 조성물 관련 특허가 공개, 등록되어 있으며, 대표적인 최근 공개 특허는 아래와 같다. 공개 번호 제2004-0083048호는 옻나무를 원료로 하는 기억력 증진과 치매치료 조성물에 관한 것이다. 공개 번호 제2004-0007174호는 옥타코사놀과 초유, 누에, 동충하초 등을 함유하는 기억력 증진 효과가 있는 특수영양식품에 관한 것이며, 공개번호 제2006-0005402호는 멜라토닌 작용제 및 니코틴 작용제를 포함하는 기억력 증진제에 관한 것이다. To date, a composition-related patent having an effect of improving a lot of memory is disclosed and registered in Korea, and a representative recently disclosed patent is as follows. Publication No. 2004-0083048 relates to memory enhancement and dementia treatment compositions based on lacquer trees. Publication No. 2004-0007174 relates to a special nutritional product having a memory-improving effect containing octacosanol and colostrum, silkworms, cordyceps and the like, and Publication No. 2006-0005402 to memory enhancing agents including melatonin agonists and nicotine agonists. It is about.
하지만, 이러한 화학 약제 및 천연 조성물이 시중에 나와 있어도, 아직은 효능이 확실한 우수한 제제가 개발되지 못하고 있는 실정이다. 또한, 시판중인 화학 약제들은 간독성, 불면증, 고혈압, 오심 구토 등의 많은 부작용을 유발하고 있다. 따라서, 부작용이 적으며, 인지 능력 저하를 예방, 치료할 수 있는 뛰어난 효과를 갖는 제제의 개발이 절실하다.However, even when such chemical agents and natural compositions are available on the market, there is still no development of excellent formulations having certain efficacy. In addition, commercially available chemicals cause many side effects such as hepatotoxicity, insomnia, hypertension, nausea and vomiting. Therefore, there is little need for the development of an agent having fewer side effects and having an excellent effect of preventing and treating cognitive decline.
본 발명은 천연물로부터 얻어지는 것으로서 인체에 부작용이 거의 없고, 콜린성 신경계 뿐 아니라, 비콜린성 신경계에도 작용하는 중증 또는 심한 인지 능력 저하로 나타나는 여러 가지 질환을 예방, 치료할 수 있는 조성물을 제공한다. The present invention provides a composition which is obtained from natural products, which has little side effects on the human body, and which can prevent and treat various diseases indicated by severe or severe cognitive decline in the cholinergic nervous system as well as in the noncholinergic nervous system.
하나의 양태로서, 본 발명은 새로운 기억력 증진 조성물에 관한 것이다. 구체적으로 본 발명은 원지, 석창포, 인삼, 당귀, 은행잎, 오미자 및 천궁으로 이루어진 혼합 추출물을 함유하는 기억력 증진 조성물에 관한 것이다. In one aspect, the present invention relates to a novel memory enhancing composition. Specifically, the present invention relates to a memory enhancing composition containing a mixed extract consisting of Wonji, Seokchangpo, Ginseng, Angelica, Ginkgo biloba, Schisandra chinensis and Cheongung.
상기 혼합 추출물의 구성 약재 중 하나인 원지(Polygala tenufolia), 석창포(Acurus gramineus), 인삼(Panax ginseng)은 뇌세포의 대사를 활성화시키면서 혈액순환을 촉진하므로 기억력 증대에 효력이 있으며, 당귀(Angelica gigas)는 데쿠신(Decursin) 혹은 데쿠시놀(Decursinol) 성분을 함유하고 있어 뇌혈류를 증가시키면서 혈관 내의 지질을 용해시키고 혈관확장으로 피의 흐름을 원활히 하는 활성이 있다. 당귀는 참당귀(Angelica gigas Nakai), 왜당귀(Angelica acutiloba Kitagawa) 또는 중국당귀(Angelica sinensis Diels)일 수 있으나, 참당귀인 것이 가장 바람직하다. 은행나무(Ginkgo biloba) 잎은 징코산(ginkoic acid) 및 징게롤(gingelol) 성분을 함유하고 있어 중추신경의 흥분과 조절작용으로 뇌신경세포의 파괴를 차단한다. 오미자(Schiandra chinensis)는 해이해지고 이완된 정신상태의 개선과 더불어 산만함 나태함을 없애주고 황홀감, 자기도취, 의지력을 강화시키는 작용이 있다. 천궁(Cnidium officinale)은 특히 여러 원인의 두통에 좋은 효과를 보이며, 통증을 제거하는 효과도 크다. 이러한 본 발명 조성물에 포함되는 각 성분들은 상기 열거한 활성 반응을 나타냄과 동시에, 서로의 효능들을 증폭시키면서 뇌혈류 촉진작용, 신생혈관 촉진효과 및 혈관 내 콜레스테롤 하강 효과 등으로 뇌기능을 향상시키는 결과를 가져온다. 이러한 본 발명의 조성물은 단 1회의 복용만으로 효력을 얻는 것이 아니라, 장시간 복용할 경우 복용하지 않은 사람들과 현저한 차이를 얻게 될 것이다. One of the constituent medicines of the mixed extract raw paper ( Polygala tenufolia ), Seokchangpo ( Acurus gramineus ) and ginseng ( Panax ginseng ) stimulate the blood circulation by activating the metabolism of brain cells, which is effective in increasing memory, Angelica gigas is decursin or decusinol ( It contains decursinol, which increases the blood flow to the blood vessels while soluble in lipids and dilates blood vessels. Angelica gigas Nakai ), Angelica acutiloba Kitagawa ) or Angelica sinensis Diels , but the most true is Angelica sinensis . Ginkgo biloba leaves contain ginkoic acid and gingelol to block brain nerve cell destruction through excitability and control of the central nervous system. Schiandra chinensis has the effect of improving the relaxed and relaxed mental state, eliminating distraction laziness and strengthening ecstasy, narcissism, and willpower. Cnidium officinale ) is particularly effective for headaches of various causes, and it is also effective in removing pain. Each of the components included in the composition of the present invention exhibits the above-described active response, and at the same time, amplifies the effects of each other, thereby improving brain function by promoting cerebrovascular flow, angiogenesis, and lowering blood vessel cholesterol. Bring. Such a composition of the present invention will not be effective in only one dose, but will be remarkably different from those who have not taken it for a long time.
더욱 상세하게, 본 발명 조성물 각각의 원료 성분 효능은 다음 아래의 문헌에 자세히 기록되어 있다. 원지는 Egashira. N. et al., Phytomedicine, 10, 467-473 (2003), Hsieh. M. et al, Phytother. Res., 14(5), 375-377 (2000), Yoshifumi Irie et al., Brain Research, 963, 282-289. Park. C. H. et al., J.Neurosci. Res., 70(3), 484-492(2002), 석창포는 Oh, M. H. et al., Phytomedicine, 11(6), 544-548 (2004), Cho, J. et al., Life Sciences, 71, 591-599 (2002), Irie et al., Brain Research, 963(1-2), 282-289(2003), 김영옥 외, 대한본초학회지, 제15권2호, 79-85(2000), 인삼은 진승하 외 Korean J. Ginseng Sci., 20(1)7-14 (1996), Hsieh. M. et al , Phytother. Res., 14(5), 375-377 (2000), Bao h. Y. et al, Arch. Pharm. Res., 28(3), 335-342 (2005), Hartley D.E., Nutr. Neurosci., 7(5-6), 325-335 (2004), 참당귀는 Kang. S. Y. et al, Neurobiol. Learning. Memory, 79(1), 11-18 (2003), Yan, J-J. et al, Prog. Neuro-Psychopharm. Biol. Psychiatry, 28(1), 52-30 (2004), Kang. S. Y. et al., J. Nat. Prod., 64(5) 683-685 (2001), 은행잎은 Gong Q-H et al., Life Sciences, 77(2), 140-148 (2005), van Dongen. M., et al., J. Clin. Epidemiol., 56(4), 367-376 (2003), Nathan. P. J. et al ., JAMA., 289(5), 546, (2003), 오미자는 Hsieh. M. et al, Phytother. Res., 13, 256-257 (1999), Nishiyama. N., et al, Biol. Pharm. Bull. 18(11). 1498-1503(1995), Kang. S. Y. et al, Life Sciences, 76(15), 1691-1705 (2005), 천궁은 Ohta. H., et al., Pharmcol. Biochem. Behavior, 45(3), 719-723 (1993), Hatip-Al-Khatib I. et al., J Pharmacol Sci. 96(1):33-41 (2004), Shen. L. X. et.al., Yao Xue Xue Bao. 37(3): 178-80 (2002). In more detail, the raw material efficacy of each of the compositions of the present invention is described in detail in the following documents. The base is Egashira. N. et al., Phytomedicine, 10, 467-473 (2003), Hsieh. M. et al, Phytother. Res., 14 (5), 375-377 (2000), Yoshifumi Irie et al., Brain Research, 963, 282-289. Park. C. H. et al., J. Neurosci. Res., 70 (3), 484-492 (2002), and Seokchang-Pho, Oh, MH et al., Phytomedicine, 11 (6), 544-548 (2004), Cho, J. et al., Life Sciences, 71 , 591-599 (2002), Irie et al., Brain Research, 963 (1-2), 282-289 (2003), Young Ok Kim et al., Korean Journal of Herbology, Vol. 15, No. 2, 79-85 (2000), Ginseng is synthesized by Jin Seung-ha et al. Korean J. Ginseng Sci., 20 (1) 7-14 (1996), Hsieh. M. et al, Phytother. Res., 14 (5), 375-377 (2000), Bao h. Y. et al, Arch. Pharm. Res., 28 (3), 335-342 (2005), Hartley D. E., Nutr. Neurosci., 7 (5-6), 325-335 (2004), Angelica is Kang. S. Y. et al, Neurobiol. Learning. Memory, 79 (1), 11-18 (2003), Yan, J-J. et al, Prog. Neuro-Psychopharm. Biol. Psychiatry, 28 (1), 52-30 (2004), Kang. S. Y. et al., J. Nat. Prod., 64 (5) 683-685 (2001), ginkgo biloba, Gong Q-H et al., Life Sciences, 77 (2), 140-148 (2005), van Dongen. M., et al., J. Clin. Epidemiol., 56 (4), 367-376 (2003), Nathan. P. J. et al., JAMA., 289 (5), 546, (2003), Schisandra chinensis. M. et al, Phytother. Res., 13, 256-257 (1999), Nishiyama. N., et al, Biol. Pharm. Bull. 18 (11). 1498-1503 (1995), Kang. S. Y. et al, Life Sciences, 76 (15), 1691-1705 (2005), Ohm. H., et al., Pharmcol. Biochem. Behavior, 45 (3), 719-723 (1993), Hatip-Al-Khatib I. et al., J Pharmacol Sci. 96 (1): 33-41 (2004), Shen. L. X. et. Al., Yao Xue Xue Bao. 37 (3): 178-80 (2002).
보다 바람직한 양태로서, 본 발명은 원지 10~30 중량%, 석창포 10~30 중량%, 인삼 5~10 중량%, 당귀 5~20 중량%, 은행잎 10~30 중량%, 오미자 10~30 중량% 및 천궁 10~30 중량%으로 이루어진 혼합 추출물을 함유하는 기억력 증진 조성물에 관한 것이다. 상기 본 발명의 혼합 추출물은 원지, 석창포 등 혼합 약재의 알코올 추출물이거나 열수 추출물이다. As a more preferred embodiment, the present invention is 10-30% by weight of raw paper, 10-30% by weight of Seokchangpo, 5-10% by weight of ginseng, 5-20% by weight of Angelica, 10-30% by weight of Ginkgo biloba, and 10-30% by weight of Schisandra chinensis. It relates to a memory enhancing composition containing a mixed extract consisting of 10-30% by weight of uterus. The mixed extract of the present invention is alcohol extract or hot water extract of mixed herbs such as raw paper, Seokchangpo.
다른 실시 형태로서, 본 발명은 원지, 석창포 등 혼합 약재의 추출물 외에, 보조 성분을 추가 함유하는 기억력 증진 조성물에 관한 것이다. 보조 성분으로는 비타민, 미네랄, 혈행 개선제가 해당된다. As another embodiment, the present invention relates to a memory enhancing composition further comprising an auxiliary component in addition to extracts of mixed medicinal herbs, such as raw paper and stone spear. Supplements include vitamins, minerals and blood circulation enhancers.
먼저, 비타민 B1, B2, B6, B12, C, 판토텐산 칼슘, 니코틴산 아미드, 엽산, 비 오틴 등의 비타민과 아연, 철, 칼슘, 마그네슘 등의 미네랄을 추가할 수 있다. 이러한 비타민 및 미네랄을 혼합 사용하면, 생체 활성 효과를 보강할 수 있으므로 바람직하다. 이때 비타민과 미네랄은 최종 조성물의 전체 중량에 대하여 0.1 내지 10 중량%로 포함되는 것이 바람직하며, 특히 비타민 B1, B2 및 C가 최종 조성물 전체 중량에 대하여 각각 0.01 내지 3 중량%, 0.01 내지 3 중량% 및 0.01 내지 4 중량%로 포함되는 것이 더욱 바람직하다. First, vitamins such as vitamins B 1 , B 2 , B 6 , B 12 , C, calcium pantothenate, nicotinic amide, folic acid, and biotin and minerals such as zinc, iron, calcium, and magnesium may be added. The use of such vitamins and minerals is preferred because it can enhance the bioactive effect. At this time, the vitamin and minerals are preferably contained in 0.1 to 10% by weight relative to the total weight of the final composition, in particular vitamins B1, B2 and C 0.01 to 3% by weight, 0.01 to 3% by weight relative to the total weight of the final composition And 0.01 to 4% by weight.
혈행 개선제는 혈행을 원활하게 하는 목적과 생체 활성 효과를 보강하기 위해서 첨가하며, 감마 리놀레익산, EPA, 토코페롤 등을 사용할 수 있다. 이때, 혈행 개선제는 토코페롤로서 최종 조성물의 전체 중량에 대하여 5 내지 30 중량%로 포함되는 것이 바람직하다. Blood circulation improving agent is added to smooth the blood circulation and to enhance the biological activity effect, gamma linoleic acid, EPA, tocopherol and the like can be used. At this time, the blood circulation improving agent is preferably included in 5 to 30% by weight based on the total weight of the final composition as tocopherol.
추가로, 본 발명의 기억력 증진 조성물은 원지, 석창포 등의 혼합 추출물, 상기 각종 비타민, 미네랄 및 혈행개선제 전체 성분을 용해하는 용매를 포함하며, 이러한 용매로는 물 또는 에틸알코올이 사용될 수 있다.In addition, the memory enhancing composition of the present invention includes a solvent for dissolving the mixed extract of raw paper, Seokchangpo, etc., the various components of the various vitamins, minerals and blood circulation improving agents, and water or ethyl alcohol may be used as such a solvent.
한편, 본 발명의 기억력 증진 조성물은 추출물을 직접 사용할 수도 있으나, 당업계에 공지된 의약품 제조방법에 따라 약학적으로 허용 가능한 담체와 함께 가공 및 제조하는 것이 바람직하다. 상기에서 담체의 예로는, 락토오즈, 덱스트로즈, 수크로즈, 솔비톨, 만니톨, 자일리톨, 에리스리톨, 말티톨, 전분, 아카시아 고무, 알지네이트, 젤라틴, 칼슘 포스페이트, 칼슘 실리케이트, 셀룰로즈, 메틸 셀룰로즈, 미정질 셀룰로즈, 폴리비닐피롤리돈, 물, 메틸 하이드록시벤조에이트, 프로필 하이드록시벤조에이트, 탈크, 마그네슘 스테아레이트 및 광물유를 들 수 있다. 또 한, 상기 기억력 증진제에는 충진제, 항응집제, 윤활제, 습윤제, 향료, 유화제 또는 방부제 등이 추가로 포함될 수 있다.On the other hand, the memory enhancing composition of the present invention may be used directly extracts, it is preferable to be processed and prepared with a pharmaceutically acceptable carrier in accordance with pharmaceutical preparation methods known in the art. Examples of the carrier in the above, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose And polyvinylpyrrolidone, water, methyl hydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate and mineral oil. In addition, the memory enhancer may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers or preservatives.
본 발명에 따른 기억력 증진 조성물은 투여 후에, 활성 성분의 신속, 지속 또는 지연된 방출을 제공할 수 있도록 당업계에 잘 알려진 방법을 사용하여 제형화할 수 있다. 제형은 정제, 알약, 과립, 분말, 새세이(sachet), 엘릭서(elixir), 현탁액, 에멀젼, 용액, 시럽, 에어로졸, 연질 또는 경질 젤라틴 캅셀, 멸균 주사용액, 멸균 분말 등의 형태일 수 있다. 과립제, 액제 등과 같은 제형으로 제조할 경우 미각을 돋우기 위하여 매실향, 레몬향, 파인애플향 또는 허브향과 같은 천연향료나 천연과즙, 클로르필린(chlorphyllin) 등의 천연색소 및 감미성분인 과당, 벌꿀, 당알콜, 설탕 등과 구연산, 구연산 나트륨과 같은 산미제를 혼합하여 사용할 수 있다.Memory enhancing compositions according to the invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration. The formulations may be in the form of tablets, pills, granules, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders and the like. When prepared in the form of granules, liquids, etc. to enhance the taste, natural flavors such as plum, lemon, pineapple or herb, or natural pigments such as fruit juice, chlorphyllin and sweeteners such as fructose, honey, Sugar alcohols, sugars and the like can be mixed with an acidulant such as citric acid and sodium citrate.
또한, 본 발명에 따른 기억력 증진 조성물은 경구, 주사 등을 포함한 여러 경로를 통하여 투여될 수 있으나, 경구 투여 방법을 이용하는 것이 가장 바람직하다.In addition, the memory enhancing composition according to the present invention can be administered through various routes including oral, injection, etc., it is most preferable to use an oral administration method.
본 발명에 따른 기억력 증진제로 사용되는 최종 조성물의 1일 복용량은 5 내지 20g의 범위이다. 그러나, 실제 복용량은 복용 방법, 환자의 연령, 성별, 체중 및 환자의 중증도 등과 같은 여러 관련 인자를 고려하여 결정되어야 하며, 상기 복용량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.The daily dose of the final composition used as memory enhancer according to the invention ranges from 5 to 20 g. However, the actual dosage should be determined in consideration of several relevant factors such as the method of administration, the age, sex, weight of the patient and the severity of the patient and the like does not limit the scope of the invention in any aspect.
본 발명의 조성물은 아래의 실시예에서 확인한 바와 같이, 뛰어난 인지 능력 개선 효과를 나타내었으며, 또한, 독성을 평가하기 위해 실시예 1-1에서 제조한 조 성물을 사용하여 ICR 마우스에서 50% 치사량 (LD50)을 측정한 결과 체중 kg당 5g이상으로 나타났으므로 본 발명의 조성물은 매우 안전함을 확인할 수 있었다. The composition of the present invention showed an excellent cognitive ability improving effect, as confirmed in the following examples, and also 50% lethal dose in ICR mice using the composition prepared in Example 1-1 to evaluate toxicity LD 50 ) was found to be more than 5g per kg body weight, it was confirmed that the composition of the present invention is very safe.
또 다른 실시 형태로서, 본 발명은 본 발명의 기억력 증진 조성물을 제조하는 방법에 관한 것이다. As another embodiment, the present invention relates to a method of preparing the memory enhancing composition of the present invention.
본 발명은 1) 원지, 석창포, 인삼, 당귀, 은행잎, 오미자 및 천궁으로 이루어진 약재를 메탄올 또는 에탄올로 40℃ 내지 90℃에서 1 내지 24시간 가열 환류 추출한 후 여과하는 단계, 2) 단계 1)에서 얻어진 여과액을 감압 건조시켜 농축 추출물을 제조하는 단계, 및 3) 단계 2)에서 얻어진 농축 추출물을 용매에 용해하는 단계를 포함하는, 본 발명의 기억력 증진 조성물을 제조하는 방법에 관한 것이다. The present invention is a step of 1) filtering the medicinal herb consisting of Wonji, Seokchangpo, Ginseng, Angelica, Ginkgo biloba, Schisandra chinensis and Cheongung after heating under reflux for 1 to 24 hours at 40 ° C to 90 ° C with methanol or ethanol, and 2) in step 1). Drying the obtained filtrate under reduced pressure to prepare a concentrated extract, and 3) dissolving the concentrated extract obtained in step 2) in a solvent.
보다 구체적으로는, 상기 각각의 천연 약재를 깨끗이 세척하고 잘게 절단한 후, 메탄올, 에탄올과 같은 저급 알코올, 바람직하게는 70-95% 에탄올을 중량 비율로 3 배 내지 8배 가하여 40℃ 내지 90℃, 바람직하게는 60℃ 내지 85℃에서, 1시간 내지 24시간, 바람직하게는 2시간 내지 6시간 동안 가열 환류 추출하고, 냉각시키지 않고 뜨거운 상태에서 여과지 또는 통상의 여과 장치를 사용하여 고형분을 제거한다. 걸러진 여과액을 회전 증발기(rotary evaoprator)나 통상의 농축기를 사용하여 감압 건조시키면 최종 알코올 추출물을 제조할 수 있다. 통상 2.0Kg의 천연물을 추출하여 감압 과정을 마치면 750g의 농축 추출물이 얻어진다. 이렇게 얻은 최종 알코올 추출물을 물 또는 에틸알코올과 같은 용매에 용해시켜 본 발명의 조성물을 제조한다. More specifically, after washing each of the natural medicines and cut finely, lower alcohols such as methanol, ethanol, preferably 70-95% ethanol by 3 to 8 times by
다르게는, 본 발명은 원지, 석창포, 인삼, 당귀, 은행잎, 오미자 및 천궁으로 이루어진 약재를 약재의 3 내지 8 배의 물로 85℃ 내지 110℃에서 1 내지 24시간 가열 환류 추출하여 여과하는 단계; 2) 단계 1)에서 여과하고 남은 고형분에 단계 1)에서 사용한 물 절반가량을 첨가하여 85℃ 내지 110℃에서 1 내지 24시간 가열 환류 추출하여 여과하는 단계; 3) 단계 1) 및 단계 2)에서 얻어진 여과액을 가열 농축하여 농축 추출물을 제조하는 단계, 및 4) 단계 3)에서 얻어진 농축 추출물을 용매에 용해하는 단계를 포함하는, 본 발명의 기억력 증진 조성물을 제조하는 방법에 관한 것이다. Alternatively, the present invention is filtered by heating reflux for 1 to 24 hours at 85 ° C to 110 ° C with medicinal herbs consisting of raw paper, Seokchangpo, Ginseng, Angelica, Ginkgo biloba, Schisandra chinensis and Cheonung; 2) adding about half of the water used in step 1) to the remaining solid after filtration in step 1) and filtering by heating under reflux for 1 to 24 hours at 85 ° C to 110 ° C; 3) preparing a concentrated extract by heating and concentrating the filtrate obtained in steps 1) and 2), and 4) dissolving the concentrated extract obtained in step 3) in a solvent. It relates to a method of manufacturing.
보다 구체적으로는, 상기 각각의 천연 약재를 깨끗이 세척한 후, 3배 내지 8배의 물을 사용하여 상기 알코올 추출방법과 동일하게 하되 85℃ 내지 110℃에서, 1시간 내지 24시간 동안, 바람직하게는 2 시간 내지 6시간 동안 1차 추출한 후, 여과하고 남은 고형분에, 다시 물을 절반량 정도를 추가하여 상기와 동일한 조건에 따라 2차 추출한다. 이 두 가지 추출액을 여과하여 고형분를 제거 한 후, 여과액을 가열 농축하여 최종 열수 추출물을 제조할 수 있다. 통상 2.0Kg의 천연물을 추출하여 감압 과정을 마치면 700g 의 농축 추출물이 얻어진다. 이렇게 얻은 최종 열수 추출물을 물 또는 에틸알코올과 같은 용매에 용해시켜 본 발명의 조성물을 제조한다. More specifically, after washing each of the natural medicinal herbs, using the water of 3 to 8 times the same as the alcohol extraction method, but at 85 ℃ to 110 ℃, preferably for 1 to 24 hours, preferably After the first extraction for 2 to 6 hours, the filtered and the remaining solid content, by adding about half the amount of water again, the second extraction under the same conditions as above. After filtering the two extracts to remove solids, the filtrate can be concentrated by heating to prepare a final hydrothermal extract. Normally, 2.0Kg of natural products are extracted and the decompression process is completed to obtain 700g of concentrated extract. The final hydrothermal extract thus obtained is dissolved in a solvent such as water or ethyl alcohol to prepare a composition of the present invention.
이하, 실시예에 의해 본 발명을 보다 구체적으로 기술한다. 그러나, 본 발명의 범위가 이들 실시예들로 제한되는 것은 아니다.Hereinafter, the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these embodiments.
실시예Example 1: 기억력 증진 조성물의 제조 1: Preparation of Memory Enhancing Composition
하기 표 1의 조성 비율에 따라 약재 원료를 구입 (경동 시장 약제상회)하여 준비하였다. According to the composition ratio of Table 1 was prepared by purchasing the pharmaceutical raw material (Kyungdong Market Pharmaceutical Co., Ltd.).
실시예Example 1-1: 알코올 추출물 제조 1-1: Alcohol Extract Manufacturer
천연 약재를 식물 전체, 잎 또는 열매를 깨끗이 세척하고 잘게 절단한 후 80% 에탄올을 중량 비율로 5배 가하여 둥근 바닥 플라스크에서 70℃에서 4시간 동안 환류 콘덴서를 부착하고 가열 환류 추출한 후, 냉각시키지 않고 뜨거운 상태에서 여과지(Whatman사 제조 No. 1001100)를 이용하여 고형분을 제거하였다. 걸러진 여과액을 통상의 농축기를 사용하여 감압 건조시켜 최종 추출물을 제조하였다. 2.0kg의 천연물을 추출하여 감압한 결과 750g의 농축 추출물을 얻을 수 있었다.After washing the whole plant, leaves or fruits with natural herbs and cutting them finely, 80% ethanol was added 5 times by weight to attach a reflux condenser for 4 hours at 70 ° C. in a round bottom flask, and refluxed under heating, without cooling. Solid content was removed using the filter paper (No. 1001100 by Whatman company) in hot state. The filtered filtrate was dried under reduced pressure using a conventional concentrator to prepare a final extract. As a result of extracting 2.0 kg of natural product and depressurizing it, 750 g of concentrated extract was obtained.
실시예Example 1-2: 1-2: 열수Hydrothermal 추출물의 제조 Preparation of Extract
4배의 물을 사용한 것을 제외하고는 상기 알코올 추출방법과 동일하게 하되 95℃에서 4시간 동안 1차 추출한 후, 여과하고 남은 고형분에 다시 물을 절반량 추가하여 상기와 동일한 조건에 따라 2차 추출하였다. 이 두 가지 추출액을 여과하여 고형분를 제거 한 후, 여과액을 가열 농축하였다. 2.0kg의 천연물을 추출하여 가열 농축한 결과 700g의 농축 추출물을 얻을 수 있었다.Except for using 4 times the water, the same as the alcohol extraction method, but the first extraction for 4 hours at 95 ℃, filtered and added to the remaining solids half the amount of the second extraction according to the same conditions as described above It was. The two extracts were filtered to remove solids, and then the filtrate was concentrated by heating. 2.0 kg of natural product was extracted and heated and concentrated to obtain 700 g of concentrated extract.
실시예Example 1-3: 보조 성분이 첨가된 조성물의 제조 1-3: Preparation of Composition Added Supplementary Components
실시예 1-1에서 제조한 알코올 추출물과 보조 성분으로 표 2에 기재한 성분을 기재된 조성비로 첨가하여 보조 성분이 첨가된 기억력 증진 조성물의 제조하였다. As the alcohol extract prepared in Example 1-1 and the auxiliary components, the components shown in Table 2 were added at the composition ratios described to prepare a memory enhancing composition to which the auxiliary components were added.
실시예Example 2: 기억력 증진 조성물의 동물 효능 시험 2: Animal efficacy test of memory enhancing composition
실시예 2-1: 인지 능력 개선 효능 평가를 위한 Y-maze 동물 실험 (Yan, J-J. et al, Prog. Neuro-Psychopharm. Biol. Psychiatry, 28(1), 52-30 (2004) 및 Rubaj, A. et al., Behavioural Brain Research, 141(1), 11-17 (2003) 참조). Example 2-1: Y-maze animal experiments for evaluating cognitive capacity improving efficacy (Yan, JJ. Et al, Prog. Neuro- Psychopharm. Biol. Psychiatry, 28 (1), 52-30 (2004) and Rubaj, A. et al., Behavioural Brain Research, 141 (1), 11-17 (2003).
본 실험은 기억 손상을 일으키는 스코폴라민(scopolamine)을 투여하여 인지능력을 저하시킨 동물과 스코폴라민 투여 후에 실시예 1-1에서 제조한 본 발명의 조성물을 투여한 동물의 운동 활성(locomotive activity)을 측정하여 비교함으로써 본 발명 조성물의 효능을 평가하는 것이다. This experiment was performed in the animals whose cognitive ability was impaired by the administration of scopolamine causing memory damage and the locomotive activity of the animals to which the composition of the present invention prepared in Example 1-1 was administered after administration of scopolamine. ) Is to evaluate the efficacy of the composition of the present invention.
2-1-1: 실험 재료 및 방법2-1-1: Experimental Materials and Methods
실험동물로는 40-43g 체중의 ICR 웅성 마우스((주)샘타코)를 구입하였다. 구입 후 일주일 동안 온도 조절 방에서 12시간/12시간, 밤/낮 주기로 안정화시켰다. 사료는 CJ 마우스 사료를 사용하였고 물은 수돗물은 자유 급식 급수하였다. 이후 마우스를 각 20마리씩 무작위로 다음의 실험군으로 나누었다. 투약을 하지 않은 대조군 그룹 20마리, 스코폴라민 (Sigma-Aldrich, 0.9% 생리 식염수 용해) 1.5mg/kg을 피하주사한 스코폴라민 실험군 그룹 20마리, 스코폴라민 투여 30분 후 실시예 1-1의 조성물 600mg/kg을 투여한 그룹 20마리로 나누었다. 경구 투여 후 30분이 지난시점에서 아래의 Y -maze 실험을 실시하였다.As an experimental animal, ICR male mouse (Samtaco) of 40-43g body weight was purchased. After purchase, it was stabilized at 12/12 hours and night / day cycles in a temperature controlled room for a week. CJ mouse feed was used as feed, and tap water was fed freely. Thereafter, 20 mice were randomly divided into the following experimental groups. 20 untreated groups, 20 scopolamine experimental groups subcutaneously injected with 1.5 mg / kg of scopolamine (Sigma-Aldrich, 0.9% physiological saline solution), Example 1- 30 minutes after scopolamine administration The animals were divided into 20 groups administered 600 mg / kg of
2-1-2: Y-maze 실험2-1-2: Y-maze experiment
순간 공간 운동 기억 행동 (immediate spatial working memory performance)을 Y-maze에서 자발적인 교체(alternation) 행동 측정 실험으로 평가하였다. Y-maze는 검은 아트지로 도배된 목재로 제작하였고, 각각의 암(arm)은 37cm이고 넓이는 5cm, 높이는 17cm이었다. 세 개의 암이 120도로 삼각을 이루고 있고, 마우스를 하나의 암에 넣어주고 8분에 걸쳐 자유롭게 각각의 암을 드나들게 하였다. 암으로의 진입횟수와 연속적으로 각각의 세 개의 암을 드나드는 교체 횟수를 수동적으로 기록하였다. 암으로의 진입은 마우스의 뒷다리가 암의 안쪽으로 완전히 진입하는 것으로 간주하였다. 교체 비율(alternation percentage)은 교체 횟수를 전체 진입 횟수 마이너스 2 값으로 나눈 숫자에 100을 곱하여 얻어진다.Instant spatial working memory performance was evaluated by spontaneous alternating behavior measurement experiments in Y-maze. The Y-maze was made of wood that was painted with black art paper, each arm was 37cm wide, 5cm wide and 17cm high. Three arms were triangulated at 120 degrees, and the mouse was placed in one arm, allowing each arm to freely move in and out over eight minutes. The number of entry into the arm and the number of replacements to and from each of the three arms in succession were recorded manually. Entry into the cancer was considered as the hind limbs of the mouse completely entering the interior of the cancer. The replacement percentage is obtained by multiplying the number by which the number of replacements is divided by the total number of entries minus two.
2-1-3: 실험 결과2-1-3: Experimental Results
Y-maze는 공간 기억을 측정하는 것으로서, 스코폴라민과 같은 콜린성 뉴런을 손상시키는 약물에 의해 인지능력이 저하될 경우, 운동 활성이 감소되어 교체 비율이 줄어드는 것으로 보고되어 있다. 즉, 총 암 진입횟수는 큰 변화가 없는 것에 반해, 연속적으로 다른 3개의 암을 들락거리는 비율이 낮아진다는 의미이다.Y-maze is a measure of spatial memory, and when cognitive decline is caused by drugs that damage cholinergic neurons such as scopolamine, it has been reported that motor activity is reduced and replacement rates are reduced. This means that the total number of cancer entry times is not significantly changed, while the rate at which three other arms enter and fall in succession is lowered.
본 실험의 결과로서 총 진입 횟수에 대해 살펴보면, 스코폴라민 처리 마우스의 암의 총 진입 횟수는 대조군 마우스와 비슷하게 나타났으며, 실시예 1-1 조성물 처리군은 증가하는 경향이 나타났다 (표 3 참조). 교체 비율에 대해 살펴보면, 스코폴라민 처리군은 교체 비율이 크게 줄어드는 결과가 나타났으나, 실시예 1-1 조성물의 처리군은 교체 비율이 유의적으로 상승하는 결과를 보였다 (표 4 및 도 1 참조). 이는 실시예 1-1 조성물이 인지 능력을 향상시키는 효과가 있음을 보여준다. As a result of the experiment, the total number of entry of the scopolamine-treated mice was similar to that of the control mice, and the Example 1-1 composition-treated group showed an increasing tendency (see Table 3). ). Looking at the replacement rate, the scopolamine treatment group was found to significantly reduce the replacement rate, but the treatment group of the composition of Example 1-1 showed a significant increase in the replacement rate (Table 4 and Figure 1 Reference). This shows that the composition of Example 1-1 has an effect of improving cognitive ability.
실시예 2-2 인지능력 개선 효능 평가를 위한 새로운 물체 인식 (Novel Object Recognition) 동물 실험 (Kang. S. Y. et al, Neurobiol. Learning. Memory, 79(1), 11-18 (2003), Vaucher,E. et al., Neurobiol. of Aging, 23(1), 87-95(2002) 참조) Example 2-2 Novel Object Recognition Animal Evaluation for Evaluation of Cognitive Improving Efficacy (Kang. SY et al, Neurobiol. Learning.Memory, 79 (1), 11-18 (2003), Vaucher, E et al., Neurobiol. of Aging, 23 (1), 87-95 (2002)).
본 발명 조성물의 기억력 증진 효과를 실험하기 위해 나무 재질의 안쪽면은 검은색으로 코팅된 박스(45cm × 45cm × 45cm)에 실험 동물을 배치하여 새로운 물체에 대한 인식 기억을 실험하였다.In order to test the memory enhancing effect of the composition of the present invention, the inner surface of the wood material was placed in a black coated box (45 cm × 45 cm × 45 cm) to test the recognition memory for a new object.
2-2-1: 실험 재료 및 방법2-2-1: Experimental Materials and Methods
실험 동물로는 40~43g 체중의 ICR 웅성 마우스((주)샘타코)를 구입 한 후 일주일 동안 온도 조절 방에서 12시간/12시간 밤/낮 주기로 안정화시켰다. 사료는 CJ 마우스 사료를 사용하였고 물은 수돗물은 자유 급식 급수하였다. 이후 마우스를 각 20마리씩 무작위로 다음의 실험군으로 나누었다. 투약을 하지 않은 대조군 그룹 20마리, 스코폴라민(Sigma-Aldrich, 0.9% 생리 식염수 용해) 1.5mg/kg을 피하주사한 스코폴라민 실험군 그룹 20마리, 스코폴라민 투여 30분 후 실시예 1-1의 조성물 600mg/kg을 투여한 그룹 20마리로 나누었다. As an experimental animal, ICR male mouse (Samtako Co., Ltd.) of 40-43 g body weight was purchased and stabilized at 12 hours / 12 hours night / day cycle in a temperature control room for one week. CJ mouse feed was used as feed, and tap water was fed freely. Thereafter, 20 mice were randomly divided into the following experimental groups. 20 untreated groups, 20 scopolamine experimental groups subcutaneously injected with 1.5 mg / kg of scopolamine (Sigma-Aldrich, 0.9% physiological saline solution), Example 1- 30 minutes after scopolamine administration The animals were divided into 20 groups administered 600 mg / kg of
2-2-2: 새로운 물체 인식 실험2-2-2: New Object Recognition Experiment
실험은 45cm × 45cm × 45cm의 정방형 박스의 정 가운데 높은 곳에 30w 백열등을 밝히고 물체를 차례로 넣어 주면서 실시하였다. 물체로는 무선 컴퓨터 마우스, 약병, 고무튜브 접은 것, 사기컵 등을 사용하였고, 대체로 크기가 비슷하며 실험용 마우스가 움직이지 못할 만큼 무거운 것을 사용하였다. 물체에서 냄새가 나는 등의 특별히 선호도가 없는 것을 선택하여 실험의 객관성을 높였다. The experiment was carried out by illuminating a 30-watt incandescent lamp in the center of a square box of 45 cm × 45 cm × 45 cm and placing the objects in sequence. The objects used were wireless computer mice, vials, rubber tube folds, fraud cups, etc., which were generally the same size and heavy enough that the experimental mouse could not move. The objectivity of the experiment was increased by selecting a thing with no particular preference such as smelling from an object.
실험 첫날에는 실험동물들을 빈 박스에 차례로 30분씩 넣어 두어 공간을 익히게 하고, 실험 둘째 날, 한 쌍의 동일한 물체를 코너에 넣어주고 다시 10분씩 적응하게 하였다. 동일한 물체에 대한 선호도를 측정하였다. 선호도는 마우스의 코나, 앞발, 얼굴 등이 물체에 접촉하는 것으로 측정하였다. 1.5cm의 근접거리에서 계속 냄새를 맡는 것도 선호도에 포함시켰다. 3시간 후 물체 한 쌍 중의 하나인 물체 A를 다른 물체 B로 바꾸어 놓고 각 물체 A, B에 접근하는(탐색하는) 시간을 기록하였다. 인식지수(Reconition Index, RI)는 (tB/(tA+tB))×100로 계산한다 (tA는 A에 접근하는 시간, tB는 B에 접근하는 시간). On the first day of the experiment, the animals were placed in an empty box for 30 minutes in order to learn the space. On the second day of the experiment, a pair of identical objects were put in the corners and adjusted for 10 minutes. The preference for the same object was measured. Preference was measured by the nose, forefoot, face, etc. of the mouse touching the object. Continued smelling at close range of 1.5cm was also included. After 3 hours, one of the pair of objects was replaced with another object B and the time to approach (search) each object A and B was recorded. Reconition Index (RI) is calculated as (tB / (tA + tB)) × 100 (tA is the time to access A, tB is the time to access B).
2-2-3: 실험 결과 2-2-3: Experimental Results
도 2에서 알 수 있듯이 대조군 그룹에 비해 스코폴라민 투여군은 현저하게 감소된 RI 값을 나타내었고, 실시예 1-1의 조성물을 투여한 실험군은 유의성 있게 RI 값이 회복하였다. As can be seen in Figure 2 compared to the control group scopolamine administration group showed a significantly reduced RI value, the experimental group administered the composition of Example 1-1 significantly recovered the RI value.
실시예 2-3: 인지능력 개선 효능 평가를 위한 수동 회피 (Passive Avoidance) 동물 실험 (Kang. S. Y. et al, Neurobiol. Learning. Memory, 79(1), 11-18 (2003) 참조) Example 2-3 Passive Avoidance Animal Experiments for Evaluating Cognitive Improving Efficacy (see Kang. SY et al, Neurobiol. Learning.Memory, 79 (1), 11-18 (2003))
수동 회피 반응이란 동물의 반응 (예를 들어 조명이 밝은 상자에서 인접한 어두운 상자로 들어서는 행동 또는 단상에서 전극이 깔린 바닥으로 내려서는 행동)이 고통스런 결과 (발바닥 전기충격과 같은)로 이어지는 경우에 일어나는 것이다. 이러한 반응-자극제시를 통해 동물은 고통스러운 결과를 초래한 반응을 회피하게 된다. Passive evasion reactions occur when an animal's reaction (for example, entering a dark box from a light box into a contiguous dark box, or descending from a single phase to the floor with electrodes) leads to a painful result (such as a foot shock). . This response-stimulation suggests that the animal avoids the reaction with painful consequences.
2-3-1: 실험 방법 및 약물 투여2-3-1: Experimental Methods and Drug Administration
실험 동물로는 5 주령의 ICR 웅성 마우스((주)샘타코)를 구입 한 후 일주일 동안 적응시켰다. 실험은 24시간 간격으로 1일 1회 실시하며 직사각형의 긴 상자를 반으로 가른 두 개의 방 (방1, 방2)이 있고, 상자 가운데에는 칸막이 문 (guillotine door)이 있으며, 전기 쇼크를 주기 위해 상자 바닥에는 그리드 (grid)를 깔았다. 휜쥐를 조명이 밝은 쪽 (방1)에 놓고 중간의 칸막이 문을 열어 쥐가 어두운 구획으로 들어갈 수 있게 하였다. 180초간 탐색 후 (방1에서) 소리와 빛과 같은 외부적 자극을 주었다. 자극을 준 후에 이를 피하려고 어두운 곳을 선호하는 흰쥐가 어두운 쪽 (방2)으로 이동하면 중간의 칸막이(길로틴)는 자동 열리게 되어 흰쥐가 어두운 방 (방2)으로 옮겨갈 수 있게 하고, 어두운 방 (방2)에서 1mA의 전기적 자극을 1초간 그리드에 흐르게 하였다. 이 학습을 4일간 시키고, 5일째에는 방1에서 방2로 옮겨가는 시간을 측정한다. 이것을 자동으로 측정하고, 학습 시행 시간 (learning trial time)이라 부른다. 이 실험의 의미는 24시간 전에 받은 쇼크를 기억하여 어두운 구획으로 들어갈 때까지의 시간으로 자동 기록하여 이 시간이 길수록 수동회피 학습과 기억이 우수한 것으로 판명하는 것이다. 즉, 방2의 전기 쇼크를 기억하기 때문에 방2로 가지 않는 지능을 시간으로 측정하는 것을 수동회피반응이라 한다. As experimental animals, five-week-old ICR male mice (Samtako Co., Ltd.) were purchased and adapted for one week. The experiment is conducted once a day at 24 hour intervals, with two rooms (
2-3-2: 실험 결과 2-3-2: Experiment Result
도 3에서 보는 바와 같이, 정상군이 9.82±0.51 초인데 비해 스코폴라민을 주사한 대조군은 6.26±0.47초로 저하되었고, 실시예 1-1의 조성물 투여군은 8.08±0.55초를 나타내어 기억력을 향상 효능이 있음을 확인할 수 있었다.As shown in FIG. 3, the control group injected with scopolamine decreased to 6.26 ± 0.47 seconds compared to the normal group of 9.82 ± 0.51 seconds, and the composition-administered group of Example 1-1 showed 8.08 ± 0.55 seconds to improve memory. This could be confirmed.
실시예 2-4: 인지능력 개선 효능 평가를 위한 수중 미로(Water Maze) 동물 실험 (Kang. S. Y. et al, Neurobiol. Learning. Memory, 79(1), 11-18 (2003) 참조) Examples 2-4: Water Maze Animal Experiments for Evaluating Cognitive Improving Efficacy (See Kang. SY et al, Neurobiol. Learning.Memory, 79 (1), 11-18 (2003))
2-4-1: 실험 재료 및 방법2-4-1: Experimental Materials and Methods
실험동물은 6주령의 웅성 위스터 래트 (Wister rat, 중앙실험동물)를 사용하였다. 실험용 쥐를 하루에 90초간 4번씩 5일간 일정한 시간에 직경 186cm의 원형수조에 넣는다. 이때, 물에 빠진 쥐는 살기 위해 3사분면에 설치된 도피대(직경 10cm)를 찾는 것을 확인할 수 있었다. 시행 각각마다 도피대를 찾는 시간을 측정하였다. 이러한 훈련을 받고 (acquisition test), 6일째 마지막 시행이 끝나면 자유수영 검사를 시행하는데 (retention test), 이때 동물들은 도피대가 제거된 채로 90초간 수영을 하게 하였다. 즉, 5일간은 도피대가 있는 채로, 6일째는 도피대가 없는 채로 실험을 실시하면 쥐는 5일간 학습한 도피대의 위치를 기억하여 도피대가 있던 3사분면으로 가려고 하고, 이렇게 가고자 하는 시간을 측정하였다. 여기서 주의할 점은 첫날부터 5일째까지의 훈련 시 쥐를 수조에 넣을 때는 매번 다른 곳에서 출발시키는 것이다. 즉, 수조의 사분면 중 매일 다른 곳에서 흰 쥐를 넣어 여러 각도에서 도피대의 위치를 찾아가는 것을 5일간 학습하는 것이다. 모든 실험 동물들의 행동을 비디오 카메라로 녹화하고, 훈련 시행에서는 출발에서부터 도피대로 올라가는데 걸린 시간을 측정하고, 90초간의 검사 시행에서는 훈련시에 도피대가 있었던 사분면에 머문 시간을 에토비젼 프로그램(Ethovision program)을 이용하여 측정하였다. The experimental animals were 6-week old male Wister rats (central test animals). The rats are placed in a round tank of diameter 186 cm at regular intervals for five days, four times for 90 seconds per day. At this time, the mice drowned in the water was found to find a shelter (diameter 10cm) installed in three quadrants to live. The time to find the escape was measured for each run. After this training (acquisition test) and at the end of the sixth day of the last run, a free swim test (retention test) was used, where the animals were allowed to swim for 90 seconds with the hides removed. That is, when the experiment was conducted with the evacuation zone for 5 days and without the evacuation zone on the 6th day, the rat remembered the position of the evacuation zone learned for 5 days and tried to go to the quadrant where the evacuation zone was located. One thing to note here is that when training rats from the first day to the 5th day, they start from different places each time. In other words, every day in the quadrant of the tank to put a white mouse in a different place to learn the location of the escape from various angles for five days. Record the behavior of all experimental animals with a video camera, measure the time taken from the start to escape to the training session, and spend the 90-second test in the quadrant where the escape was during training. ) Was measured.
2-4-2: 실험 결과 2-4-2: Experimental Results
스코폴라민(1mg/kg)을 주사하고 실시예 1-1의 조성물을(500mg/kg) 투여한 쥐가 스코폴라민(1mg/kg)만을 주사한 쥐보다 빨리 도피대에 도달하였다. 실시예 1-1의 조성물을 투여한 쥐의 경우, 마지막 6일째 18시간 후에는 정상 쥐가 도피대에 도달하는 시간과 거의 동일함을 알 수 있었다. 구체적으로 아래의 표에서 알 수 있는 바와 같이, 6일째 획득시간의 평균값은 정상쥐의 경우는 9.33±0.76, 스코폴라민 단독 투여군의 경우는 15±1.32, 스코폴라민과 본 발명의 조성물을 투여한 쥐의 경우는 11±1.32로서, 본 발명 조성물을 투여한 쥐는 정상쥐와 비슷한 결과를 보였다. 따라서 본 발명 조성물의 공간 인지기억 학습수행 능력에 대한 증진 효과를 확인할 수 있었다.Mice injected with scopolamine (1 mg / kg) and administered with the composition of Example 1-1 (500 mg / kg) reached the escape zone faster than mice injected with scopolamine (1 mg / kg) only. In the rats to which the composition of Example 1-1 was administered, it was found that after 18 hours on the last 6 days, the rats were almost the same as the time to reach the escape zone. Specifically, as can be seen in the table below, the average value of the acquisition time on
실시예Example 2-5: 인지능력 개선 효능 평가를 위한 신경세포 손상 방어효과 동물 실험 2-5: Animal Experiments on Neuroprotective Effects of Neuronal Cell Damage Assessment
2-5-1: 실험 재료 및 방법2-5-1: Experimental Materials and Methods
실험동물로는 6주령의 웅성 위스터 래트 (Wister rat, 중앙실험동물)를 이용하였다. 상기 래트의 뇌로 올라가는 4개의 동맥혈을 폐색시켜서 혈액이 흐르지 못하게 허혈을 유발하였다. 즉, 2개의 척추동맥을 결찰하고 다른 2개의 총경동맥을 결찰하였다. 허혈을 유발한 후, 생리식염수(500mg/kg)를 투여한 그룹과, 실시예 1-1의 조성물(500mg/kg)을 투여한 군으로 나누었다. 1주일 후에 각 쥐의 뇌를 획출하여 해마를 포함한 부위를 조직 절편하여 크레실 바이올렛(cresyl violet)으로 염색하여 단위면적당 생존한 신경세포수를 계수하였다. 동물 당 3개의 조직에서 양측 뇌 부위를 관찰하였으며 3명의 관찰자가 실험군에 대한 정보를 모르게 하고 세포수를 계수하였다. 이들을 평균한 수치를 실험값으로 하였다. As the test animals, 6-week-old male Wister rats (central test animals) were used. 4 arterial blood clots going up to the rat brain were blocked, causing ischemia to prevent blood flow. That is, two vertebral arteries were ligated and the other two common carotid arteries. After inducing ischemia, the group was divided into a group administered with saline (500 mg / kg) and a group administered with the composition (500 mg / kg) of Example 1-1. One week later, the brains of each rat were extracted, tissue sections containing the hippocampus were stained with cresyl violet, and the number of surviving neurons per unit area was counted. Two brain regions were observed in three tissues per animal, and three observers were informed of the experimental group and counted. The numerical value which averaged these was made into the experiment value.
2-5-2: 실험 결과 2-5-2: Experimental Results
정상군의 뇌의 세포수는 단위면적당 340±8.6 세포/mm2이고, 4개의 동맥혈을 폐색시켜 뇌허혈을 유발한 후, 생리적 식염수를 주사한 쥐의 단위 면적당 살아있는 뇌의 세포수는 101±10 세포/mm2이며, 4개의 동맥혈을 폐색시켜 뇌허혈을 유발한 후, 실시예 1-1의 조성물을 주사한 쥐의 단위 면적당 살아있는 뇌의 세포수는 245.8±9.8 세포/mm2였다. 따라서, 본 발명 조성물의 백분율 효능은 [(245.8-101) / (340-101)] X 100으로서 60.6%의 유의한 방어 효과를 나타내었다.The number of brain cells in the normal group is 340 ± 8.6 cells / mm 2 per unit area, and the number of living brain cells per unit area of rats injected with physiological saline after blockage of four arterial blood induces cerebral ischemia. / mm 2 , and after occluding 4 arterial blood to induce cerebral ischemia, the number of living brain cells per unit area of the rat injected with the composition of Example 1-1 was 245.8 ± 9.8 cells / mm 2 . Thus, the percentage efficacy of the composition of the present invention showed a significant protective effect of 60.6% as [(245.8-101) / (340-101)]
또한, 도 5는 상기 실험에서 크레실 바이올렛으로 염색한 조직 절편인 CA1 추체세포 및 신경의 사진을 나타낸다. 도 5의 A는 정상군의 추체세포 사진이고, 대부분의 신경들은 정상적인 형태를 유지함을 확인할 수 있었다 (도 5의 B). 허혈유발 후 생리식염수 처리군에서의 추체세포는 약하게 보이고, 해마의 CA1 부분의 신경세포가 손상을 많이 받았으며 (도 5의 C), 신경세포는 고사가 고사되어 글라이오시스를 동반한 세포상해가 있음을 확인할 수 있었다 (도 5의 D). 반면, 허혈유발 후 실시예 1-1의 조성물 처리군에서는 해마 CA1 추체세포의 손상이 상당히 줄어들었음을 확인하였다 (도 5의 F와 G). 체온의 변화는 허혈유발 생리식염수 투여 쥐와 본 발명 조성물을 투여한 쥐 어디에도 없었으므로 본원 발명 조성물의 신경 보호 효과는 체온 저하에 따른 것이 아님을 알 수 있었다. In addition, Figure 5 shows a picture of the CA1 cone cells and nerve tissue sections stained with cresyl violet in the experiment. 5A is a picture of a vertebral cell of a normal group, and it was confirmed that most nerves maintained a normal shape (B of FIG. 5). After the ischemic induction, the vertebral cells in the saline treatment group appeared weak, and the nerve cells in the CA1 part of the hippocampus were damaged a lot (FIG. 5C), and the neurons were killed by apoptosis. It was confirmed that the (Fig. 5D). On the other hand, in the composition treatment group of Example 1-1 after ischemia induction, it was confirmed that the damage of hippocampal CA1 vertebral cells was significantly reduced (FIG. 5F and G). Since there was no change in body temperature in the ischemic physiological saline-administered rats and the rats to which the composition of the present invention was administered, it was found that the neuroprotective effect of the composition of the present invention was not due to the decrease in body temperature.
실시예Example 3. 인지능력 개선 효능 평가를 위한 뇌파 측정을 통한 임상실험 3. Clinical trial through EEG measurement for evaluation of cognitive improvement effect
3-1: 뇌파 측정 3-1: EEG measurement
본 임상 실험은 피검자 성인 20인 학생 20인을 대상으로 (주)Laxtha의 QEEG-2 뇌파 측정기를 이용한 학습능력 측정 소프트웨어를 사용하여 실시하였다. 본 실험은 본 발명 조성물의 집중력, 인지능력, 인지 강도, 종합 학습능력 향상 효과를 측정하는 실험이다. 각 피검자에게 실시예 1-1의 조성물을 하루 10g씩 2개월간 투여한 후, 본 실험에서 사용한 뇌파 측정기의 매뉴얼을 이용하여 뇌파측정을 하였다. This clinical trial was conducted on 20 adult students of 20 subjects using learning ability measurement software using Laxtha's QEEG-2 EEG test system. This experiment is to measure the concentration, cognitive ability, cognitive strength, comprehensive learning ability improvement effect of the present invention composition. Each subject was administered 10 g of the composition of Example 1-1 for 2 months per day, and then EEG was measured using the manual of the EEG test instrument used in this experiment.
3-2: 뇌파를 이용한 학습 능력 측정3-2: Learning ability measurement using brain waves
신경생리학적 학습능력 검사 및 뇌파-바이오피드백 훈련용 학습능력 소프트웨어(LXSMD 3-1)를 이용하여 잠재적 학습 능력을 진단하였다. 사용한 소프트웨어는 상기의 (주)Laxtha의 뇌파 측정기와 연동되어 작동하고, 인지강도, 인지속도, 집중력, 작업부하도, 좌/우뇌 활성에 해당하는 신경생리학적 지표 추출하여 뇌파에 의한 잠재적 학습능력을 진단하게 된다. 소프트웨어의 매뉴얼에 따라 잠재적 학습 능력을 진단하였다.Potential learning ability was diagnosed using the Neurophysiological Learning Test and EEG-Biofeedback Training Learning Software (LXSMD 3-1). The software used works in conjunction with Laxtha's EEG test and extracts neurophysiological indicators that correspond to cognitive intensity, cognitive speed, concentration, workload, and left / right brain activity to detect potential learning ability by EEG. Diagnosis is made. Potential learning skills were diagnosed according to the software manual.
3-3: 실험 결과 3-3: Experimental Results
피검자 성인 20인 학생 20인을 각각 대상으로 한 인지강도, 집중력, 학습능력수준을 아래의 표 6 내지 9에 나타내었다. 상기 실험 결과로서 알 수 있듯이, 본 발명의 조성물은 사람에 있어서도 유의적으로 인지강도, 집중력 및 학습능력 수준을 향상시켰고, 학생 및 성인 모두에게 효과를 나타냄을 확인할 수 있었다. Cognitive intensity, concentration, and learning ability levels for 20
상기 살펴본 바와 같이, 원지, 석창포, 인삼, 당귀, 은행잎, 오미자 및 천궁으로 이루어진 혼합 추출물을 함유하는 본 발명의 조성물은 인체에 부작용이 없으며, 뛰어난 기억력 향상 효과를 나타낸다. 따라서, 본 발명의 조성물은 치매를 비롯한 인지 능력 저하로 나타내는 여러 가지 질환을 예방하거나 치료할 수 있다. As described above, the composition of the present invention containing a mixed extract consisting of Wonji, Seokchangpo, Ginseng, Angelica, Ginkgo biloba, Schisandra chinensis and Cheongung has no side effects on the human body and shows an excellent memory-improving effect. Therefore, the composition of the present invention can prevent or treat various diseases indicated by cognitive decline including dementia.
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2012047063A2 (en) * | 2010-10-08 | 2012-04-12 | 제일약품주식회사 | Pharmaceutical composition for aiding dementia treatment or treating dementia, containing angelica gigas nakai extract only, ginkgo leaf extract only, or mixture of angelica gigas nakai extract and ginkgo leaf extract |
| CN103055014A (en) * | 2012-12-31 | 2013-04-24 | 青岛华仁信息技术开发有限公司 | Folium ginkgo health product for preventing senile dementia and preparation method thereof |
| CN103235029A (en) * | 2013-03-25 | 2013-08-07 | 江苏省质量安全工程研究院 | Chinese herbal processing process identification method based on sparse recognition algorithm and time-of-flight mass spectrometry |
| KR101505870B1 (en) * | 2014-07-29 | 2015-03-25 | 대한약품공업 주식회사 | Composition for improving or treating memory loss due to degenerative dementia and preparation method thereof |
| CN105166896A (en) * | 2015-09-06 | 2015-12-23 | 吉林大学 | Health-care food capable of improving memory in auxiliary manner |
| KR20190111225A (en) * | 2018-03-22 | 2019-10-02 | 주식회사 메타포뮬러 | A composition for preventing or treating cognitive impairment comprising an omega3 fatty acid |
| KR102089490B1 (en) * | 2018-11-15 | 2020-03-16 | 대한민국(농촌진흥청장) | Composition for improving memory or composition for preventing and treating Alzheimer's dementia containing boiled silkworm products having silk protein and angelica extracts |
| KR20200087606A (en) * | 2019-01-11 | 2020-07-21 | 장영용 | Compositions for Improving Memory Power and Learning Ability, and Relaxing stress |
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| KR101486041B1 (en) * | 2014-03-31 | 2015-01-23 | 주식회사 자연인 | Korean herbal aromatic composition for improving memory and method for manufacturing the same |
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| KR20000006625A (en) * | 1998-07-20 | 2000-02-07 | 묵인희 | Treatment For Dementia Comprising Ginseng Extract |
| KR100623164B1 (en) * | 2004-07-22 | 2006-09-19 | 재단법인서울대학교산학협력재단 | A method for preparing purified extract for the prevention and treatment of stroke and ischemia from ginkgo biloba leaves extract and the composition containing the same |
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| WO2012047063A2 (en) * | 2010-10-08 | 2012-04-12 | 제일약품주식회사 | Pharmaceutical composition for aiding dementia treatment or treating dementia, containing angelica gigas nakai extract only, ginkgo leaf extract only, or mixture of angelica gigas nakai extract and ginkgo leaf extract |
| WO2012047063A3 (en) * | 2010-10-08 | 2012-07-19 | 제일약품주식회사 | Pharmaceutical composition for aiding dementia treatment or treating dementia, containing angelica gigas nakai extract only, ginkgo leaf extract only, or mixture of angelica gigas nakai extract and ginkgo leaf extract |
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| KR101505870B1 (en) * | 2014-07-29 | 2015-03-25 | 대한약품공업 주식회사 | Composition for improving or treating memory loss due to degenerative dementia and preparation method thereof |
| CN105166896A (en) * | 2015-09-06 | 2015-12-23 | 吉林大学 | Health-care food capable of improving memory in auxiliary manner |
| KR20190111225A (en) * | 2018-03-22 | 2019-10-02 | 주식회사 메타포뮬러 | A composition for preventing or treating cognitive impairment comprising an omega3 fatty acid |
| KR102089490B1 (en) * | 2018-11-15 | 2020-03-16 | 대한민국(농촌진흥청장) | Composition for improving memory or composition for preventing and treating Alzheimer's dementia containing boiled silkworm products having silk protein and angelica extracts |
| KR20200087606A (en) * | 2019-01-11 | 2020-07-21 | 장영용 | Compositions for Improving Memory Power and Learning Ability, and Relaxing stress |
| KR20220159536A (en) * | 2021-05-25 | 2022-12-05 | 제천한약영농조합법인 | Manufacturing method of Medical Herb Tea for Preventing and Alleviating Dementia |
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