KR102059452B1 - Composition comprising the extract of Patriniae Radix for preventing and treating Attention Deficit Hyperactivity Disorder - Google Patents
Composition comprising the extract of Patriniae Radix for preventing and treating Attention Deficit Hyperactivity Disorder Download PDFInfo
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- KR102059452B1 KR102059452B1 KR1020180052595A KR20180052595A KR102059452B1 KR 102059452 B1 KR102059452 B1 KR 102059452B1 KR 1020180052595 A KR1020180052595 A KR 1020180052595A KR 20180052595 A KR20180052595 A KR 20180052595A KR 102059452 B1 KR102059452 B1 KR 102059452B1
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Abstract
본 발명은 주의력결핍 과잉행동장애(ADHD) 개선 및 치료용 조성물에 관한 것으로, 본 발명의 패장근 추출물을 유효성분으로 함유하는 조성물은 신경전달물질 재흡수 활성을 억제하여 직관, 충동성, 대인관계, 정서변화, 집중력, 억제력과 같은 과잉행동장애 등을 효과적으로 예방, 개선 또는 치료할 수 있고, 도파민, 노르에피네프린, 세로토닌 신경계에 대하여 다양한 차별적 선택성을 지녀 다중 타겟을 탄력적으로 조절할 수 있으므로 단일 타겟으로서 의존성, 성장저해 및 수면장애와 같은 부작용을 최소화할 수 있으며, 천연물로부터 유래된 것이어서 체내에 심각한 자극을 가한다거나 유해한 작용을 유발함이 없이 안전하게 지속적으로 사용할 수 있다.The present invention relates to a composition for improving and treating attention deficit hyperactivity disorder (ADHD), the composition containing the extract of the root muscle of the present invention as an active ingredient by inhibiting the neurotransmitter reuptake activity intuition, impulse, interpersonal relationship It can effectively prevent, improve or treat hyperactivity disorders such as emotional changes, concentration, and suppression.Also, it is possible to flexibly control multiple targets with various differential selectivity for dopamine, norepinephrine, and serotonin nervous system. Side effects such as growth inhibition and sleep disorders can be minimized, and since they are derived from natural products, they can be safely and continuously used without causing serious irritation or harmful effects in the body.
Description
본 발명은 주의력결핍과잉행동장애(ADHD) 개선 및 치료용 조성물에 관한 것이다.The present invention relates to a composition for improving and treating attention deficit hyperactivity disorder (ADHD).
주의력결핍과잉행동장애(Attention Deficit Hyperactivity Disorder, ADHD)는 뇌 내 신호전달 체계 이상으로 나타나는 뇌발달 장애이다. ADHD는 5-10%의 높은 유병률을 보이고 있으나, 그 원인인자 및 병리 기전이 명확하게 규명되지 않은 뇌신경 분야의 난치성 질환 중 하나로 여겨지고 있다. 질환은 조기 아동기에 진단되는 경우가 많지만, 최근 연구에서는 뒤늦게 발병되는 성인 ADHD 발병에 대한 보고도 다수 존재한다(Adgnew-Blais et al., 2016; Caye et al., 2016; Faraone and Biederman, 2016; Moffitt et al 2015). 전세계적으로 아동 및 청소년의 경우 약 2.2-7.2 %, 성인의 경우 2.8-5.0 % 의 유병률을 나타내며 (Fayyad et al., 2007, 2016; Polanczyk et al. 2007; Thomas et al., 2015; Willcutt, 2012) 주 증상으로 과도하고 지속적인 부주의, 과잉행동 및 충동적 행동 (Inattention, hyperactivity, impulsivity)을 나타낸다(American Psychiatric Association, 2000). 이는 인지, 사회적 상호 작용, 고용, 자기 관리 및 이동성과 같은 영역에서 기능 장애 또한 초래하며 유병률이 높고, 이후 정신병 동반 질환과의 강한 상관 관계, 성인기까지의 지속성 및 평생 동안의 기능 장애로 인해 ADHD는 전 세계적으로 중요한 장애로 인식되고 있다(Fayyad et al., 2016).Attention Deficit Hyperactivity Disorder (ADHD) is a brain development disorder that manifests itself in the brain's signaling system. Although ADHD has a high prevalence of 5-10%, it is considered to be one of the refractory diseases in the field of cerebral nerve where its causative agent and pathology are not clearly identified. The disease is often diagnosed early in childhood, but recent studies also report a number of late onset ADHD cases (Adgnew-Blais et al., 2016; Caye et al., 2016; Faraone and Biederman, 2016; Moffitt et al 2015). Prevalence rates around 2.2-7.2% for children and adolescents and 2.8-5.0% for adults worldwide (Fayyad et al., 2007, 2016; Polanczyk et al. 2007; Thomas et al., 2015; Willcutt, 2012) Main symptoms include excessive and persistent inattention, hyperactivity, and impulsivity (Aatan Psychiatric Association, 2000). It also leads to dysfunction in areas such as cognition, social interaction, employment, self-management and mobility, and has a high prevalence, and because of its strong correlation with psychotic illness, persistence to adulthood, and lifetime dysfunction, It is recognized as an important disorder around the world (Fayyad et al., 2016).
현재 사용되는 약물들로는 메틸페니데이트(methylphenidate), 아토목세틴(atomoxetine) 등이 있는데 이들은 단일 타겟으로서 의존성, 성장저해 및 수면장애와 같은 심각한 3대 부작용을 초래하므로 사용에 주의를 기하고 있고 환자나 보호자들에게 사용이 기피되고 있다.Currently used drugs include methylphenidate and atomoxetine, which are single targets and cause serious side effects such as dependency, growth inhibition and sleep disorders. Its use is being avoided by guardians.
연간 7조원에 달하는 ADHD 치료제 시장에도 불구하고 현 치료제는 25%에 달하는 치료 불응 환자군의 존재와 다양한 부작용으로 인해 효능과 안전성을 확보한 획기적인 신약 개발이 절실하다.Despite the 7 trillion won annual ADHD drug market, the current drug is in need of new drug development that has secured efficacy and safety due to the presence of 25% of non-treated patients and various side effects.
ADHD의 증상이나 병인과 밀접한 연관성을 지닌 직관, 충동성, 대인관계, 정서변화, 집중력, 억제력 등은 도파민, 노르에피네프린, 세로토닌 신경계와 서로 복잡한 관련성을 보이고, ADHD 환자의 경우 개인에 따라 위의 각 증상이 발현되는 정도가 다르다.Intuition, impulsivity, interpersonal relationships, emotional changes, concentration, and suppression, which are closely related to the symptoms and etiology of ADHD, have complex relationships with the dopamine, norepinephrine, and serotonin nervous systems. The degree of symptoms is different.
본 발명에서 제시하는 패장근(敗醬根)은 녹장근으로도 불리는 식물로서 뚝갈 혹은 마타리과의 뿌리를 칭한다. 사포닌과 유사한 성분인 파트리니오시드는 용혈작용 및 국소자극작용효과를 가지고 주로 대장질환, 해독작용, 출산후유증, 전립선치료, 항염작용, 피부질환, 치질 및 신경쇠약증에 사용된다고 알려져 있다.Pajanggeun (敗 醬 根) proposed in the present invention refers to the roots of the family Dungalgal or Matariaceae as a plant also referred to as kungjanggeun. It is known that saponin-like ingredient, patlinioside, has hemolytic and local stimulating effects and is mainly used for colon disease, detoxification, postpartum sequelae, prostate treatment, anti-inflammatory, skin disease, hemorrhoids and nervous breakdown.
관련 특허로는, 폐암치료용 조성 및 건강기능식품(KR 2011-0027371), 뇌암예방 및 치료(KR 2011-0012222), 췌장암 치료조성 및 건강기능식품(KR 2011-0085596) 등이 있으나, 패장근 추출물의 과잉행동개선 용도와 관련된 연구는 전무한 상태이다.Related patents include composition and health functional food for treating lung cancer (KR 2011-0027371), prevention and treatment of brain cancer (KR 2011-0012222), pancreatic cancer treatment composition and health functional food (KR 2011-0085596), etc. There is no research related to the use of extracts in improving overaction.
본 발명자들은 도파민, 노르에피네프린, 세로토닌 신경계에 대해서 다양한 차별적 선택성을 지닌 물질 개발을 통해서 다중 타겟을 탄력적으로 조절 가능하고, 초기 개발단계부터 부작용을 최소화 하면서 ADHD를 예방, 개선 또는 치료할 수 있는 천연물을 개발하고자 노력하였다. 그 결과, 패장근 추출물의 도파민, 노르에피네프린, 세로토닌 수송체의 다중 신경전달물질 재흡수 활성억제효과, 마우스 모델에서의 과잉행동 억제효과를 확인함으로써, 본 발명을 완성하였다. The inventors have developed a natural product that can flexibly control multiple targets through the development of substances with various differential selectivities for dopamine, norepinephrine, and serotonin nervous system, and prevent, improve or treat ADHD while minimizing side effects from the initial development stage. I tried to As a result, the present invention was completed by confirming the effect of inhibiting dopamine, norepinephrine, and serotonin transporter's multiple neurotransmitter reuptake activity, and the inhibitory effect on the mouse model.
따라서, 본 발명의 목적은 패장근(Patriniae radix) 추출물을 유효성분으로 함유하는 주의력결핍과잉행동장애(Attention Deficit Hyperactivity Disorder, ADHD)의 예방, 개선 및 치료용 조성물을 제공하는데 있다.Accordingly, it is an object of the present invention to provide a composition for the prevention, improvement and treatment of Attention Deficit Hyperactivity Disorder (ADHD), which contains Patriniae radix extract as an active ingredient.
상술한 과제를 해결하기 위하여 본 발명은 패장근(Patriniae radix) 추출물을 유효성분으로 함유하는 주의력결핍과잉행동장애(Attention Deficit Hyperactivity Disorder, ADHD)의 예방 및 치료용 약학적 조성물을 제공한다.In order to solve the above problems, the present invention provides a pharmaceutical composition for the prevention and treatment of Attention Deficit Hyperactivity Disorder (ADHD), containing the extract of Patriniae radix as an active ingredient.
또한, 본 발명은 패장근(Patriniae radix) 추출물을 유효성분으로 함유하는 주의력결핍과잉행동장애(ADHD)의 예방 및 개선용 건강기능식품을 제공한다.In addition, the present invention provides a dietary supplement for the prevention and improvement of attention deficit hyperactivity disorder (ADHD) containing the extract of Patriniae radix as an active ingredient.
본 발명의 패장근 추출물을 유효성분으로 함유하는 조성물은 신경전달물질 재흡수 활성을 억제하여 직관, 충동성, 대인관계, 정서변화, 집중력, 억제력과 같은 과잉행동장애 등을 효과적으로 예방, 개선 또는 치료할 수 있는 우수한 효과가 있다.The composition containing the extract of Paengeungeun as an active ingredient effectively inhibits, improves or treats hyperactivity disorders such as intuition, impulse, interpersonal relationship, emotional change, concentration, and inhibitory ability by inhibiting neurotransmitter reuptake activity. It has an excellent effect.
또한, 본 발명의 패장근 추출물을 유효성분으로 함유하는 조성물은 도파민, 노르에피네프린, 세로토닌 신경계에 대하여 다양한 차별적 선택성을 지녀 다중 타겟을 탄력적으로 조절할 수 있으므로 단일 타겟으로서 의존성, 성장저해 및 수면장애와 같은 부작용을 최소화할 수 있다.In addition, the composition containing the extract of the root muscle of the present invention as an active ingredient has a variety of differential selectivity to the dopamine, norepinephrine, serotonin nervous system can be elastically controlled multiple targets, such as dependency, growth inhibition and sleep disorders as a single target Side effects can be minimized.
또한, 본 발명의 패장근 추출물은 동물실험 결과 독성을 나타내지 않고 천연물로부터 유래된 것이어서, 과잉행동장애의 예방, 개선, 치료 효능 이외에 체내에 심각한 자극을 가한다거나 유해한 작용을 유발함이 없이 안전하게 지속적으로 사용할 수 있다.In addition, the extract of Pajanggeun of the present invention is derived from natural products without showing toxicity as a result of animal experiments, and safely and continuously without causing serious irritation or harmful effects in the body in addition to the prevention, improvement and treatment efficacy of hyperactivity disorder. Can be used.
도 1은 HEK-293-세포주에서 패장근 추출물에 의한 각 신경전달물질 수송체의 재흡수활성 억제효과를 나타낸 결과로, 도 1a는 도파민 신경전달물질 재흡수활성 억제효과, 도 1b는 노르에피네프린 신경전달물질 재흡수활성 억제효과, 도 1c는 세로토닌 신경전달물질 재흡수활성 억제효과를 나타낸 도면이다.
도 2는 실험동물에서 패장근 추출물에 의한 과잉행동 억제효과를 확인한 결과로, A는 주의력결핍/과잉행동장애 모델(SHR)에서 과잉행동 억제효과를 확인한 결과이고, B는 대조군인 위스타-교토랫드(WKY) 과잉행동을 확인한 결과이다.
도 3은 실험동물에 케타민을 투여하여 수면을 유도하고 패장근 추출물에 의한 수면 유도 및 수면 지속시간을 확인한 도면으로, A는 입면시간, B는 수면 지속시간을 확인한 결과이다.
도 4는 패장근 추출물의 독성을 확인한 것으로, 도 4a는 일주일 몸무게 변화를 확인한 것이고, 도 4b는 장기의 독성 사진을 나타낸 것이며, 도 4c는 장기무게 변화를 확인한 것이다.1 is a result of inhibiting the reuptake activity of each neurotransmitter transporter by the extract of the root muscle in HEK-293-cell line, Figure 1a is the inhibitory effect of dopamine neurotransmitter reuptake activity, Figure 1b is norepinephrine neurons Delivery material reuptake inhibitory effect inhibitory effect, Figure 1c is a view showing the inhibitory effect of serotonin neurotransmitter reuptake activity.
Figure 2 is a result of confirming the inhibitory effect of the excess activity by the extract of the root muscle in the experimental animals, A is the result of confirming the inhibitory effect of hyperactivity in the attention deficit / hyperactivity disorder model (SHR), B is a control group Wistar-Kyoto This is the result of checking the excess behavior of rat (WKY).
3 is a diagram showing the induction of sleep by the administration of ketamine to the experimental animal and the sleep induction and sleep duration by the extract of the root muscle, A is the sleep time, B is the result of confirming the sleep duration.
Figure 4 is to confirm the toxicity of the extract of the root, Figure 4a is to confirm the weight change of the week, Figure 4b is a picture showing the toxicity of the organ, Figure 4c is to confirm the change in organ weight.
이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.
본 발명의 일 양태에 따르면, 본 발명은 패장근(Patriniae radix) 추출물을 유효성분으로 함유하는 주의력결핍과잉행동장애(Attention Deficit Hyperactivity Disorder, ADHD)의 예방, 개선 및 치료용 조성물을 제공한다.According to one aspect of the invention, the present invention provides a composition for the prevention, improvement and treatment of Attention Deficit Hyperactivity Disorder (ADHD) containing an extract of Patriniae radix as an active ingredient.
본 발명자들은 천연 추출물을 이용한 부작용 및 독성이 없는 ADHD 치료제를 개발하기 위하여 예의 연구 노력한 결과, 패장근 추출물의 도파민, 노르에피네프린, 세로토닌 수송체의 재흡수 활성 억제효과 및 ADHD 동물모델에서의 과잉행동 억제효과를 확인하였으며, 패장근 추출물을 경구 투여한 실험동물에서 수면장애 및 세포독성의 부작용이 없음을 확인하였다.The present inventors have diligently studied the development of ADHD therapeutics without side effects and toxicity using natural extracts. As a result, the effect of inhibiting the reuptake activity of dopamine, norepinephrine, and serotonin transporter of Pasengeun extract and hyperactivity suppression in ADHD animal model The effect was confirmed, and there were no side effects of sleep disorders and cytotoxicity in the experimental animals administered orally the myofascimus extract.
본 명세서에서 "패장근 추출물"이란 천연물인 패장근으로부터 유효성분을 추출하여 얻어진 것이라면 특별히 한정하지 않고 모두 포함한다. 예컨대, 패장근을 물이나 유기용매에 넣고 정치, 교반, 가압 또는 가열 등의 수단을 통해 유효성분을 용출함으로써 얻어진 결과물을 들 수 있다. 또한, 그와 같이 하여 얻어진 액상 추출물을 동결건조함으로써 얻어진 동결건조물을 포함한다. 또한, 그러한 동결건조물을 분쇄한 분말을 포함한다. 그 외에도 유효성분을 추출하기 위해 가능한 수단이 무엇이든지 가리지 않고 추출된 모든 추출물을 포함하며, 추출된 후 동결 건조 등의 가공을 거친 것까지도 모두 포함된다. 기타, 중탕이나 상온에 의한 추출법과 같이 예로부터 전해 내려오거나, 한의서 또는 교과서에 기재되어 있는 통상적인 추출법에 의한 추출물을 포함한다. 또한 특정 활성 화합물들을 분리하기 위한 특별한 추출법이나 각종 칼럼 크로마토그래피 방법 등을 통하여 얻어진 분획 추출물을 포함한다.In the present specification, "peanus root extract" includes all of them without particular limitation, as long as it is obtained by extracting an active ingredient from natural root melancholia. For example, the result obtained by putting a package root in water or an organic solvent and eluting an effective component through means, such as standing, stirring, pressurization, or heating, is mentioned. It also includes a lyophilized product obtained by lyophilizing the liquid extract thus obtained. It also includes a powder obtained by grinding such lyophilisate. In addition, any possible means for extracting the active ingredient includes all the extracted extracts, including all the processed after extraction and freeze-drying. Others include extracts obtained by conventional extraction methods such as extraction by baths or room temperature, or by conventional extraction methods described in Chinese medicine or textbooks. In addition, the fraction extract obtained through a special extraction method or various column chromatography methods for separating the specific active compounds.
본 발명의 일 구현예에 있어서, 본 발명의 패장근 추출물은 패장근에 용매를 첨가한 후 초음파를 처리하여 수득한 것일 수 있다. 상기 용매로는 약학적으로 허용되는 유기용매라면 어느 것을 사용해도 무방하며, 물 또는 유기용매를 사용할 수 있으며, 이에 제한되지는 않으나, 예를 들어, 정제수, 메탄올(methanol), 에탄올(ethanol), 프로판올(propanol), 이소프로판올(isopropanol), 부탄올(butanol) 등을 포함하는 탄소수 1 내지 4의 알코올, 아세톤(acetone), 에테르(ether), 벤젠(benzene), 클로로포름(chloroform), 에틸아세테이트(ethyl acetate), 메틸렌 클로라이드(methylene chloride), 헥산(hexane) 및 시클로헥산(cyclohexane) 등의 각종 용매를 단독으로 혹은 혼합하여 사용할 수 있다. 바람직하게는 에탄올(주정)을 사용할 수 있으며, 보다 바람직하게는 80~95 w/v% 에탄올을 사용하는 것이 좋다.In one embodiment of the present invention, the mesothelioma extract of the present invention may be obtained by treating with ultrasonic waves after adding a solvent to the melancholia. As the solvent, any pharmaceutically acceptable organic solvent may be used, and water or an organic solvent may be used, but is not limited thereto. For example, purified water, methanol, ethanol, C1-C4 alcohol, acetone, ether, benzene, chloroform, ethyl acetate, including propanol, isopropanol, butanol, etc. ), Methylene chloride, hexane and cyclohexane may be used alone or in combination. Preferably ethanol (alcohol) can be used, more preferably 80 to 95 w / v% ethanol is used.
본 발명의 일 구현예에 있어서, 본 발명의 패장근 추출물은 80~ 95 w/v% 에탄올을 이용하여 40~50 ℃에서 10 ~ 20 분 동안 초음파 처리 후 1~3 시간 정치를 하루 10회씩 3일간 반복하여 수득한 것일 수 있다.In one embodiment of the present invention, the mycorrhizal extract of the present invention using 80 ~ 95 w / v% ethanol for 10 to 20 minutes at 40 ~ 50 ℃ for 10 to 20
본 발명의 일 구현예에 있어서, 상기 초음파 처리에서 초음파의 주파수는 30~40 kHz일 수 있고, 바람직하게는 35~37 kHz 일 수 있고, 가장 바람직하게는 36.5 kHz 일 수 있다.In one embodiment of the present invention, the frequency of the ultrasonic wave in the ultrasonic treatment may be 30 ~ 40 kHz, preferably 35 ~ 37 kHz, most preferably 36.5 kHz.
본 발명의 패장근 추출물은 단순히 에탄올 용매를 이용하여 추출한 추출물과 비교하여 추출 효율이 증대되며, 더불어 과잉행동 억제 활성에서 더욱 우수한 효과를 보인다.The extract of Pajanggeun of the present invention is increased in extraction efficiency compared to the extract extracted with simply ethanol solvent, and also shows a more excellent effect on the hyperactivity inhibiting activity.
본 발명에 따른 약제학적 조성물은 약제학적 조성물의 제조에 통상적으로 사용하는 적절한 담체, 부형제 및 희석제를 더 포함할 수 있다.The pharmaceutical composition according to the present invention may further comprise suitable carriers, excipients and diluents commonly used in the manufacture of pharmaceutical compositions.
본 발명의 조성물의 약학적 투여 형태는 이들의 약학적 허용가능한 염의 형태로도 사용될 수 있고, 또한 단독으로 또는 타 약학적 활성 화합물과 결합뿐만 아니라 적당한 집합으로 사용될 수 있다. 상기 염으로는 약학적으로 허용되는 것이면 특별히 한정되지 않으며, 예를 들어 염산, 황산, 질산, 인산, 불화수소산, 브롬화수소산, 포름산 아세트산, 타르타르산, 젖산, 시트르산, 푸마르산, 말레산, 숙신산, 메탄술폰산, 벤젠술폰산, 톨루엔술폰산,나프탈렌술폰산 등을 사용할 수 있다.The pharmaceutical dosage forms of the compositions of the present invention may be used in the form of their pharmaceutically acceptable salts, and may be used alone or in combination with other pharmaceutically active compounds as well as in a suitable collection. The salt is not particularly limited as long as it is pharmaceutically acceptable. For example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, hydrofluoric acid, hydrobromic acid, formic acid acetic acid, tartaric acid, lactic acid, citric acid, fumaric acid, maleic acid, succinic acid, methanesulfonic acid , Benzene sulfonic acid, toluene sulfonic acid, naphthalene sulfonic acid and the like can be used.
본 발명에 따른 약제학적 조성물은, 각각 통상의 방법에 따라 산제, 과립제, 정제, 캡슐제, 현탁액, 에멀젼, 시럽, 에어로졸 등의 경구형 제형, 연고, 크림 등의 외용제, 좌제 및 멸균 주사용액 등을 비롯하여 약제학적 제제에 적합한 어떠한 형태로든 제형화하여 사용될 수 있다.The pharmaceutical composition according to the present invention may be used in the form of powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, oral formulations, ointments, creams, external preparations, suppositories, and sterile injectable solutions. It may be used in formulated in any form suitable for pharmaceutical formulations.
본 발명에 따른 조성물에 있어서, 본 발명의 추출물의 바람직한 투여량은 대상자의 연령, 성별, 체중, 증상, 질병의 정도, 약물형태, 투여경로 및 기간에 따라 다르지만, 당업자에 의해 적절하게 선택될 수 있다. 그러나, 바람직한 효과를 위해서, 본 발명의 조성물은 1일 0.001 내지 500 mg/kg 체중으로 투여하는 것이 좋다. 투여는 하루에 한번 투여할 수도 있고, 수회 나누어 투여할 수도 있다. 또한, 그 투여량은 연령, 성별, 체중, 질병의 정도, 투여경로 등에 따라서 증감될 수 있다. 따라서, 상기 투여량은 어떠한 면으로든 본 발명의 범위를 한정하는 것은 아니다.In the compositions according to the invention, the preferred dosage of the extract of the invention depends on the subject's age, sex, weight, symptoms, extent of disease, drug form, route of administration and duration, but may be appropriately selected by those skilled in the art. have. However, for the desired effect, the composition of the present invention is preferably administered at 0.001 to 500 mg / kg body weight per day. Administration may be administered once a day or may be divided several times. In addition, the dosage may be increased or decreased depending on age, sex, weight, degree of disease, route of administration, and the like. Therefore, the above dosage does not limit the scope of the present invention in any aspect.
본 발명에 따른 조성물은, 쥐, 생쥐, 가축, 인간 등의 포유동물에 비경구, 경구 등의 다양한 경로로 투여될 수 있으며, 투여의 모든 방식은 예상될 수 있는데, 예를 들면, 경구, 직장 또는 정맥, 근육, 피하, 자궁내 경막 또는 뇌혈관내(intracerebroventricular) 주사에 의해 투여될 수 있다.The composition according to the present invention can be administered to mammals such as rats, mice, livestock, humans by various routes, such as parenteral, oral, and all modes of administration can be expected, for example, oral, rectal Or by intravenous, intramuscular, subcutaneous, intrauterine dural or intracerebroventricular injection.
한편, 본 발명에 따른 조성물은, 심각한 독성 및 부작용은 없으므로 예방 목적으로 장기간 사용 시에도 안심하고 사용할 수 있다.On the other hand, the composition according to the present invention, there is no serious toxicity and side effects can be used with confidence even for long-term use for the purpose of prevention.
본 발명에 의한 조성물은, 유효성분으로서 추출물을 전체 조성물 중량 중 0.001 내지 99.9중량% 함유할 수 있다. 0.001중량% 이상이어야 효과를 기대할 수 있고, 불순물의 존재 등으로 인해 99.9중량%를 넘기 어렵기 때문이다.The composition according to the present invention may contain 0.001 to 99.9% by weight of the extract as an active ingredient in the total weight of the composition. This is because the effect can be expected to be 0.001% by weight or more, it is difficult to exceed 99.9% by weight due to the presence of impurities.
한편, 경구 투여의 목적으로 본 발명의 추출물을 정제, 캡슐, 츄잉정, 작은 봉지, 과립, 분말, 액체 용액, 현탁액, 분산액, 에멀젼, 시럽 등의 제제로 제형화하는 경우에 있어서는, 아라비아 고무, 옥수수 전분, 미세 결정질 셀룰로오스 또는 젤라틴과 같은 결합제, 인산 이칼슘 또는 락토오즈와 같은 부형제, 알긴산, 옥수수 전분 또는 감자 전분과 같은 붕해제, 스테아르산 마그네슘과 같은 윤활제, 슈크로오즈 또는 사카린과 같은 감미제 및 페퍼민트, 메틸 살리실산염 또는 과일향과 같은 향미제가 포함될 수 있으며, 투여 단위 형이 캡슐제인 경우에는 상기 성분 외에도 폴리에틸렌 글리콜 또는 지방유와 같은 액상 담체가 포함될 수 있다.On the other hand, in the case of formulating the extract of the present invention into tablets, capsules, chewing tablets, small bags, granules, powders, liquid solutions, suspensions, dispersions, emulsions, syrups and the like for the purpose of oral administration, gum arabic, Binders such as corn starch, microcrystalline cellulose or gelatin, excipients such as dicalcium phosphate or lactose, disintegrants such as alginic acid, corn starch or potato starch, lubricants such as magnesium stearate, sweeteners such as sucrose or saccharin and Flavoring agents such as peppermint, methyl salicylate or fruit flavors may be included, and when the dosage unit form is a capsule, liquid carriers such as polyethylene glycol or fatty oil may be included in addition to the above components.
또한, 본 발명은 추출물 및 식품학적으로 허용 가능한 식품보조 첨가제를 포함하는 건강 기능성 식품 조성물을 제공한다. 본 발명에 따른 식품 조성물은, 예를 들어, 츄잉껌, 캐러멜 제품, 캔디류, 빙과류,과자류 등의 각종 식품류, 청량 음료, 미네랄 워터, 알코올 음료 등의 음료 제품, 비타민이나 미네랄 등을 포함한 건강기능성 식품류일 수 있다.The present invention also provides a health functional food composition comprising an extract and a food acceptable additive. Food composition according to the present invention, for example, chewing gum, caramel products, candy, ice cream, various foods such as confectionery, soft drinks, mineral water, beverage products such as alcoholic beverages, health functional foods including vitamins and minerals, etc. Can be.
이 때, 상기 식품 중의 상기 추출물의 양은 전체 식품 중량의 0.001 내지 99.9 중량%로 가할 수 있으며, 음료 중에는 100 ㎖를 기준으로 0.001 내지 0.1 g, 더 바람직하게는 0.05 내지 0.1 g의 비율로 가할 수 있다.At this time, the amount of the extract in the food may be added to 0.001 to 99.9% by weight of the total food weight, in the beverage may be added in a ratio of 0.001 to 0.1 g, more preferably 0.05 to 0.1 g based on 100 ml. .
본 발명의 추출물을 함유하는 음료는 지시된 비율로 필수 성분으로서 상기 추출물 또는 화합물 유도체를 함유하는 외에는 다른 성분에는 특별한 제한이 없으며 통상의 음료와 같이 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다.The beverage containing the extract of the present invention is not particularly limited to other ingredients except for containing the extract or the compound derivative as essential ingredients in the indicated proportions, and contains various flavors or natural carbohydrates as additional ingredients, such as ordinary drinks. can do.
상기 외에 본 발명의 건강기능성 식품 조성물은 여러 가지 영양제, 비타민, 광물(전해질), 합성 풍미제 및 천연 풍미제 등의 풍미제, 착색제 및 증진제(치즈, 초콜릿 등), 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 기능성 식품 조성물들은 천연 과일 쥬스 및 과일 쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율은 그렇게 중요하진 않지만 본 발명의 조성물 100 중량부 당 0 내지 약 20 중량부의 범위에서 선택 되는 것이 일반적이다.In addition to the above, the functional food composition of the present invention includes various nutrients, vitamins, minerals (electrolytes), synthetic flavors such as synthetic and natural flavors, colorants and enhancers (such as cheese and chocolate), pectic acid and salts thereof, alginic acid And salts thereof, organic acids, protective colloid thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the functional food compositions of the present invention may contain fruit flesh for the production of natural fruit juices and fruit juice beverages and vegetable beverages. These components can be used independently or in combination. The proportion of such additives is not so critical but is usually selected in the range of 0 to about 20 parts by weight per 100 parts by weight of the composition of the present invention.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 보다 구체적으로 설명하기 위한 것으로, 본 발명의 범위가 이들 실시예에 의해 제한되지 않는다는 것은 당업계에서 통상의 지식을 가진 자에 있어서 자명할 것이다.Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention more specifically, it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.
실시예 1. 패장근 추출물의 준비Example 1.Preparation of Root Root Extract
본 발명에서 사용한 패장근 추출물은 한국생명공학연구원의 한국식물추출물은행으로부터 공급받아 실험에 사용하였으며, 본 발명의 패장근 추출물은 패장근 분말 100~110 g을 에탄올 1L를 사용하여 45 ℃에서 15분 동안 36.5 kHz 의 주파수로 초음파처리(sonication)하고(SD-ULTRASONIC CLEANER, SDN-900H) 2시간 정치하기를 하루에 10번씩 3일간 시행하여 추출한 생약시료이다. 추출한 물질은 무형광 솜을 여과지로 이용하여 여과하였고, 45 ℃에서 Rotary Evaporator, N-1000SWD (EYELA)를 이용하여 농축하였다. 농축된 시료는 -70 ℃ 에서 24시간 동안 Biotron coporation의 Modul spin 40을 이용하여 건조하였다. 건조된 추출물은 원 시료로서 -4 ℃ 저온실에 보관하였다.The extract of Paengeun root used in the present invention was supplied from Korea Plant Extract Bank of Korea Research Institute of Bioscience and Biotechnology and used for experiments. It is a herbal sample extracted by sonication (SD-ULTRASONIC CLEANER, SDN-900H) at a frequency of 36.5 kHz for 10
실험예 1. 세포주에 패장근 추출물을 처리하여 도파민, 노르에피네프린, 세로토닌 수송체의 신경전달물질 재흡수 억제효과 확인Experimental Example 1. Determination of neurotransmitter reuptake inhibitory effect of dopamine, norepinephrine, serotonin transporter by treatment with extract of Pa root muscle
HEK-293 세포주에 수송체 cDNA를 형질주입(transfection)한다. Transient transfection의 경우 pCDNA 3.1 (Zeo)에 subcloning 된 수송체 cDNA (3 μg/100mm dish, PEI: 10 μl)를 100 mm 컬쳐 디쉬에 형질주입한다. 다음날 24 웰 플레이트에 세포를 다시 시딩하였다. 이후 배지를 제거하고 버퍼에 희석한 패장근 추출물을 각 웰당 200 μl 가하고 37 ℃에서 15 분 동안 인큐베이션 하였다. 이후 20-30 nM [3H]-DA(도파민)을 포함하는 100 μl 버퍼를 넣고 37 ℃에서 5분간 인큐베이션 하였다. NE과 5-HT의 경우에는 NE 또는 5-HT 수송체 cDNA를 형질전환 시킨 세포를 사용하며, 기질로서는 [3H]-NE(노르에피네프린) 또는 [3H]-5-HT(세로토닌)를 사용하였다(도 1). 재빨리 반응액을 제거하고 1 ml 차갑게 식힌 버퍼로 3번 세척한 후, 1% SDS 0.5 ml 로 녹인 다음 활성도를 측정하여 확인하였다.The transporter cDNA is transfected into the HEK-293 cell line. For transient transfection, the transfected cDNA (3 μg / 100 mm dish, PEI: 10 μl) subcloned in pCDNA 3.1 (Zeo) is transfected into a 100 mm culture dish. The next day the cells were seeded again in 24 well plates. Thereafter, the medium was removed and 200 μl of each of the root muscle extracts diluted in the buffer was added and incubated at 37 ° C. for 15 minutes. Then 100 μl buffer containing 20-30 nM [3H] -DA (dopamine) was added and incubated at 37 ° C. for 5 minutes. In the case of NE and 5-HT, cells transformed with NE or 5-HT transporter cDNA were used, and [3H] -NE (norepinephrine) or [3H] -5-HT (serotonin) was used as a substrate. (FIG. 1). The reaction solution was quickly removed, washed three times with 1 ml cold chilled buffer, dissolved in 0.5 ml of 1% SDS, and then measured for activity.
도파민 재흡수 억제효능을 실험한 결과, vehicle 대조군에 비해 도파민 재흡수 억제제로서 알려진 GBR12909(vanoxerine) 인 양성대조군은 약 85%정도의 억제효능을 보였고 패장근은 이와 비슷한 효능을 나타내었다 (도 2a). 노르에피네프린 재흡수 억제효능에 대해 실험 결과, 양성대조군과 유사하게 약 80% 정도 노르에피네프린 재흡수 억제효능을 나타내었다 (도 2b). 세로토닌 재흡수 억제효능에 대한 실험결과, 양성대조군인 벤라팍신(venlafaxine)은 약 60%의 세로토닌 재흡수 억제효능을 보였고 패장근은 대조군에 비해 약 90% 억제 효능을 보인 바 양성 대조군보다 약 30% 정도 더 좋은 억제 효능을 나타내었다 (도 2c).As a result of experiments on the inhibitory effect of dopamine reuptake, GBR12909 (vanoxerine) positive control group, which is known as dopamine reuptake inhibitor, showed about 85% inhibitory effect compared to vehicle control group. . Experimental results on the inhibitory effect of norepinephrine resorption showed that norepinephrine reuptake inhibitory effect similar to the positive control group (Fig. 2b). As a result of the serotonin reuptake inhibitory effect, venlafaxine, a positive control, showed about 60% of serotonin reuptake inhibitory effect, and the myofascissus showed about 90% inhibition effect compared to the control group. Better inhibitory efficacy was shown (FIG. 2C).
실험예 2. 실험동물에서 패장근 추출물에 의한 과잉행동 억제효과 확인(도 2의 A, B)Experimental Example 2. Confirmation of the inhibitory effect of the excessive action by the extract of the root muscle in the experimental animal (A, B of Figure 2)
3주령 주의력결핍 과잉행동장애(ADHD)모델인 본태성 고혈압쥐(SHR)와 대조군인 위스타-교토랫드(WKY)에게 패장근 추출물 50 mg/kg 또는 Vehicle 10% tween80을 일주일 동안 경구투여 하고, 넓은 박스안에 10분 동안 움직인 거리 및 시간을 측정하는 개방장소 탐색활동도 측정실험(Open Field Test, OFT)을 통해 과잉행동 억제효과를 확인하였다.50 mg / kg or 10
ADHD 동물모델인 SHR에서의 실험결과, 투여 첫째 날에 vehicle보다 패장근 추출물 투여군에서 350 cm, 40초간 덜 움직이는 것으로 나타났고, 투여 7일차는 vehicle보다 패장근 추출물 투여군에서 400 cm, 50초간 덜 움직이는 것으로 나타났다(도 2의 A).Experimental results in SHR, the animal model of ADHD, showed 350 cm and 40 seconds less movement in the PA group than the vehicle on the first day of administration. On the 7th day, 400 cm and 50 seconds less in the PA group than the vehicle. (A in FIG. 2).
대조군인 위스타-교토랫드(WKY) 실험결과, 투여 첫째날에는 두 그룹간 유의적 차이를 보이지 않았고 투여 7일차는 패장근 추출물 투여군에서 100 cm 덜 움직였지만 총 움직인 시간변화는 없는 것으로 나타났다(도 2의 B).The control group, Wistar-Kyoto rat (WKY), showed no significant difference between the two groups on the first day of administration. 2, B).
실험예 3. 실험동물에서 패장근 추출물에 의한 수면장애 부작용 확인(도 3)Experimental Example 3. Confirmation of the sleep disorder side effects by the extract of Pa root muscle in the experimental animal (Fig. 3)
4주령 생쥐에 패장근 추출물 50 mg/kg 또는 길초근 추출물(수면의 양성대조군, 10 mg/kg)을 일주일 동안 경구투여 방법으로 섭취시킨 후 5주령이 된 생쥐에 케타민 100 mg/kg을 복강주사하여 수면을 유도하였다. 케타민을 처리한 생쥐가 배를 보이고 뒤집혀 있으면 잠이 든 것이고, 배를 보이고 뒤집혀 있던 생쥐가 다시 등을 보이는 자세를 바꾸면 수면에서 완전히 깨는 것으로 간주하였다.Four-week-old mice were ingested 50 mg / kg of Pa root muscle extract or Geuncho root extract (positive control of sleep, 10 mg / kg) by oral administration for one week, followed by intraperitoneal injection of
패장근 추출물에 의한 수면 유도 및 수면 지속시간 효과를 확인한 결과, 수면유도시간 및 지속시간에 대해 음성대조군과 차이가 없었으므로 수면유도효과, 수면시간 증진효과 혹은 수면장애에 대한 영향은 없는 것으로 확인되었다(도 3).As a result of confirming the effect of sleep induction and sleep duration by the extract of Paengeun root, it was confirmed that there was no difference in sleep induction time and duration as compared with the negative control group, so there was no effect on sleep induction effect, sleep time enhancement effect or sleep disorder. (FIG. 3).
실험예 4. 패장근 추출물의 독성 확인(도 4)Experimental Example 4. Confirmation of the toxicity of the extract of Paengeuneun (Fig. 4)
4주령 생쥐에 패장근 추출물 50 mg/kg을 7일 동안 주입하는 기간 동안의 무게변화를 측정한 결과, 그룹간의 체중변화는 없는 것으로 확인되었다(도 4의 A). 7일 후 5주령 생쥐에서 뇌, 폐, 심장, 가슴샘, 간, 신장, 비장, 정소를 적출하여 장기무게 변화를 측정한 결과, 통계적으로 유의한 변화는 나타나지 않았다(도 4의 B, C).As a result of measuring the weight change during the period of injecting 50 mg / kg of Pa root muscle extract in 4 weeks old mice, it was confirmed that there was no weight change between the groups (FIG. 4A). After 7 days, the brain, lung, heart, thymus, liver, kidney, spleen, and testis were extracted from the 5 week-old mice, and the change in organ weight was not found. No statistically significant changes were observed (Fig. 4B and C).
약물 투여 일주일 동안 대조군, 패장근 추출물 군, 양성대조군에 대한 몸무게 변화, 장기무게 변화, 장기의 변화의 차이가 없었으므로 투여 농도에 대한 독성이 없음을 확인할 수 있었다.There was no difference in body weight, organ weight, and organ changes for the control group, the extract of Pa root muscle, and the positive control group for one week.
Claims (6)
상기 패장근 추출물은 패장근 초음파 추출물인 것을 특징으로 하는, 조성물.The method of claim 1,
The patch root extract is characterized in that the ultrasonic wave extract, composition.
상기 패장근 초음파 추출물은 80~ 95 w/v% 에탄올을 이용하여 40~50 ℃에서 10 ~ 20 분 동안 초음파 처리 후 1~3 시간 정치를 하루 10회씩 3일간 반복하여 수득한 것을 특징으로 하는, 조성물.The method of claim 2,
The ultrasonic wave extract of Pajanggeun is obtained by repeating 1 to 3 hours of standing 10 times a day for 3 days after sonication for 10 to 20 minutes at 40 to 50 ℃ using 80 ~ 95 w / v% ethanol, Composition.
상기 패장근 추출물의 유효량은 0.001~500 mg/kg 인 것을 특징으로 하는, 조성물.The method of claim 1,
The effective amount of the mycorrhizal extract is characterized in that 0.001 ~ 500 mg / kg, the composition.
상기 건강기능식품은 음료류, 육류, 초코렛, 식품류, 과자류, 피자, 라면, 기타 면류, 껌류, 사탕류, 아이스크림류, 알코올 음료류, 비타민 복합제 및 건강보조식품류로 이루어진 군으로부터 선택되는 것을 특징으로 하는, 건강기능식품.The method of claim 5,
The health functional food is characterized in that selected from the group consisting of beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements, Nutraceutical.
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