KR20050051737A - An extract of chrisanthemi sibirici herba having an anti-allergic effect - Google Patents
An extract of chrisanthemi sibirici herba having an anti-allergic effect Download PDFInfo
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- 239000000284 extract Substances 0.000 title claims abstract description 57
- 230000003266 anti-allergic effect Effects 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 208000026935 allergic disease Diseases 0.000 claims abstract description 13
- 238000009835 boiling Methods 0.000 claims abstract description 10
- 235000013376 functional food Nutrition 0.000 claims abstract description 8
- 238000011282 treatment Methods 0.000 claims abstract description 7
- 239000013566 allergen Substances 0.000 claims description 27
- 230000028327 secretion Effects 0.000 claims description 21
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 18
- 230000000903 blocking effect Effects 0.000 claims description 9
- 239000000843 powder Substances 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 239000004480 active ingredient Substances 0.000 claims description 5
- 239000012141 concentrate Substances 0.000 claims description 5
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- 239000012153 distilled water Substances 0.000 claims description 3
- 239000000203 mixture Substances 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
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- 230000001988 toxicity Effects 0.000 abstract 1
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- 108090000695 Cytokines Proteins 0.000 description 2
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 2
- 239000007990 PIPES buffer Substances 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 201000009961 allergic asthma Diseases 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
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- 102000005962 receptors Human genes 0.000 description 2
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- 230000009885 systemic effect Effects 0.000 description 2
- IHPYMWDTONKSCO-UHFFFAOYSA-N 2,2'-piperazine-1,4-diylbisethanesulfonic acid Chemical compound OS(=O)(=O)CCN1CCN(CCS(O)(=O)=O)CC1 IHPYMWDTONKSCO-UHFFFAOYSA-N 0.000 description 1
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 description 1
- 108010041986 DNA Vaccines Proteins 0.000 description 1
- 229940021995 DNA vaccine Drugs 0.000 description 1
- 206010012434 Dermatitis allergic Diseases 0.000 description 1
- 208000004262 Food Hypersensitivity Diseases 0.000 description 1
- 206010016946 Food allergy Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 102000009438 IgE Receptors Human genes 0.000 description 1
- 108010073816 IgE Receptors Proteins 0.000 description 1
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- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 208000016599 Uterine disease Diseases 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 229940125715 antihistaminic agent Drugs 0.000 description 1
- 239000000739 antihistaminic agent Substances 0.000 description 1
- 201000008937 atopic dermatitis Diseases 0.000 description 1
- 210000003651 basophil Anatomy 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000006549 dyspepsia Diseases 0.000 description 1
- 239000000469 ethanolic extract Substances 0.000 description 1
- 235000020932 food allergy Nutrition 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 229960001340 histamine Drugs 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 208000000509 infertility Diseases 0.000 description 1
- 230000036512 infertility Effects 0.000 description 1
- 231100000535 infertility Toxicity 0.000 description 1
- 150000002617 leukotrienes Chemical class 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 229940124595 oriental medicine Drugs 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009962 secretion pathway Effects 0.000 description 1
- 231100000872 sexual dysfunction Toxicity 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000003809 water extraction Methods 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
- A61K36/287—Chrysanthemum, e.g. daisy
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
- A23V2200/304—Foods, ingredients or supplements having a functional effect on health having a modulation effect on allergy and risk of allergy
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/20—Natural extracts
- A23V2250/21—Plant extracts
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/50—Methods involving additional extraction steps
- A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
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- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
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Abstract
본 발명은 항알레르기 효과를 가지는 구절초(Chrisanthemi sibirici Herba) 추출물에 관한 것으로, 보다 구체적으로는, 전통한약재인 구절초로부터 열탕 또는 유기용매를 이용하여 항알레르기 효과를 가지는 성분을 추출하는 방법 및 상기 추출된 물질을 함유하는 항알레르기 기능성 식품 또는 의약조성물에 대한 것이다.The present invention relates to gujeolcho ( Chanthanthemi sibirici Herba ) extract having an antiallergic effect, and more specifically, a method of extracting an ingredient having an antiallergic effect from boiling water or an organic solvent from gujeolcho which is a traditional Chinese medicine and the extracted Anti-allergic functional foods or pharmaceutical compositions containing the substance.
본 발명에 따르면, 기존에 알레르기 치료의 증상을 완화하는 접근법에서 그 근본 원인을 치료함으로서 우수한 항알레르기 효과를 가질뿐더러 독성과 부작용 없는 기능성 식품 또는 의약품으로 유용하게 활용될 것으로 기대된다.According to the present invention, by treating the root cause in the conventional approach to alleviate the symptoms of allergy treatment, it is expected to have a good anti-allergic effect and to be useful as a functional food or medicine without toxicity and side effects.
Description
본 발명은 항알레르기 효과를 가지는 구절초(Chrisanthemi sibirici Herba) 추출물에 관한 것으로, 보다 구체적으로는, 전통한약재인 구절초로부터 열탕 또는 유기용매를 이용하여 항알레르기 효과를 가지는 성분을 추출하는 방법 및 상기 추출된 물질을 함유하는 항알레르기 기능성 식품 또는 의약조성물에 대한 것이다.The present invention relates to gujeolcho ( Chanthanthemi sibirici Herba ) extract having an antiallergic effect, and more specifically, a method of extracting an ingredient having an antiallergic effect from boiling water or an organic solvent from gujeolcho which is a traditional Chinese medicine and the extracted Anti-allergic functional foods or pharmaceutical compositions containing the substance.
비만세포 및 혈중 호염구는 여러 가지 알레르기 질환 즉, 알레르기성 비염, 알레르기성 피부염, 천식, 음식 알레르기 및 아나필락틱 쇼크 등을 유발하는 체내 세포로 알려져 있다. 이들 세포는 세포표면에 알레르기를 유발하는 항체인 IgE에 대한 수용체(FcεRI)를 가지고 있고, 그것은 알레르기를 유발하는 물질(항원 혹은 알러젠으로 불리워짐)에 의해 자극을 받아 자신이 가지고 있는 다양한 알레르기를 유발시키는 물질을 세포 바깥으로 분비한다. Mast cells and blood basophils are known to be cells in the body causing various allergic diseases, such as allergic rhinitis, allergic dermatitis, asthma, food allergy and anaphylactic shock. These cells have receptors for IgE (FcεRI), antibodies that cause allergies to the cell surface, which are stimulated by allergens (called antigens or allergens) to induce various allergies they have Secrete the substance outside the cell.
알레르기를 치료하는 다양한 방법들이 존재하며, 대부분의 현대의 알레르기 치료는 그 원인을 없애기보다는 증상을 완화하는 방향으로 연구가 진행되어지고 있다. 대표적으로 알러젠에 의해 비만세포 등에서 분비된 히스타민이나 류코트리엔 등의 수용체에 대한 길항약들이 주를 이루고 이러한 약물들이 거대한 시장을 이루고 있다. 그러나 이러한 약물은 환자에게 투여 후 단기간 내에 내성을 보이기 때문에 일정기간 지난 후 혹은 반복 투여시 환자들의 증상을 호전시키지 못하는 경우가 많다. 따라서 본 발명에서는 비만세포 등에서 알레르기 물질분비를 원천적으로 차단함으로서 환자에게서 일어나는 항히스타민제등의 부작용이 없는 알레르기 치료제를 개발하고자 하였다. There are many ways to treat allergies, and most modern allergy treatments are being studied to alleviate the symptoms rather than eliminate the cause. Representatively, antagonists of histamine and leukotriene, which are secreted by mast cells from allergens, are dominant and these drugs form a huge market. However, since these drugs are resistant to patients within a short period of time after administration, they often do not improve symptoms after a certain period of time or repeated administration. Therefore, the present invention was intended to develop an allergy treatment without side effects such as antihistamines occurring in patients by blocking allergen secretion in mast cells and the like.
이 외에 다른 치료 방법으로 알레르기 환자가 앓고 있는 알레르기에 대한 알러젠을 규명 후 이를 소량씩 수년간 투여하여 그 알레르기를 점차 감소시키는 방법이 있다. 하지만 이 방법은 치료기간이 우선 수년이 걸리고, 아나필락틱 쇼크 등을 유발시킬 수 있다는 단점이 있다. 다음으로 DNA백신을 사용하는 방법, IgE가 비만세포의 수용체에 결합하는 것을 차단하는 치료법, 그리고 알레르기를 유발 하는 사이토카인인 IL4에 대한 항체 치료법등의 치료적 접근법이 있다. 하지만 이러한 접근법들은 비용이 많이 들거나 아직 완전히 그 치료효과가 규명되지 않았다는 점이 있다. In addition to other treatments, allergens for allergy sufferers to identify allergies and then administered in small amounts for several years there is a way to gradually reduce the allergy. This method, however, has the disadvantage that treatment takes several years and can cause anaphylactic shock. Next, there are therapeutic approaches such as the use of DNA vaccines, therapies that block IgE from binding to mast cell receptors, and antibodies to IL4, an allergic cytokine. However, these approaches are expensive or have not yet been fully elucidated.
이에 본 발명자들은 알레르기성 비염, 천식 등 인간에게 불편함을 주거나 치명적인 다양한 알레르기를 유발하는 비만세포로부터 알레르기 유발물질의 분비를 차단함으로서 그 질환을 치료할 수 있는 물질을 비교적 안전성이 보장되어 있는 전통한방약재로부터 확보하고자 예의 노력한 결과, 천연물로부터 오랫동안 사용해오던 다양한 한방약재를 선택하여 실험해 오던 중 구절초의 에탄올 추출물이 저 농도에서 시험관에서 비만세포로부터 알레르기 유발물질의 분비를 억제한다는 것을 확인하고, 알레르기 동물모델에서 그 증상을 억제한다는 것을 증명하여 본 발명을 완성하게 되었다. 본 발명을 완성하게 되었다. Accordingly, the present inventors block the secretion of allergens from mast cells that cause discomfort to humans such as allergic rhinitis and asthma or cause various fatal allergens. As a result of diligent efforts to ensure that the ethanol extract of gujeolcho inhibits the secretion of allergens from mast cells in vitro at low concentrations, while selecting various herbal medicines that have been used for a long time from natural products, allergic animal models The present invention was completed by demonstrating that the symptoms are suppressed. The present invention has been completed.
결국 본 발명의 주된 목적은 알레르기성 비염, 천식 등 인간에게 불편함을 주거나 치명적인 다양한 알레르기를 유발하는 비만세포로부터 분비된 알레르기 유발물질의 분비를 차단하는 구절초 추출물을 제공하는데 있다.After all, the main object of the present invention is to provide a Gujeolcho extract that blocks the secretion of allergens secreted from mast cells causing various allergic or fatal allergies such as allergic rhinitis and asthma.
본 발명의 다른 목적은 항알레르기 효과를 가지는 구절초 추출물의 제조방법을 제공하는데 있다.It is another object of the present invention to provide a method of preparing Gujeolcho extract having an antiallergic effect.
본 발명의 또 다른 목적은 상기 구절초 추출물을 유효성분으로 함유하는 알레르기성 질환 치료용 조성물 및 기능성 식품을 제공하는데 있다. Still another object of the present invention is to provide a composition and functional food for treating allergic diseases containing Gujeolcho extract as an active ingredient.
상기 목적을 달성하기 위하여, 본 발명은 알레르기 유발물질의 분비를 차단하여 항알레르기 효과를 나타내는 구절초(Chrisanthemi sibirici Herba) 추출물을 제공한다.In order to achieve the above object, the present invention provides an anti-allergic ( Chanthanthemi sibirici Herba ) extract exhibiting an antiallergic effect by blocking the secretion of allergens.
본 발명은 또한, (a) 구절초를 증류수로 세척하여 분쇄기로 분쇄하는 단계; (b) 상기 분쇄된 구절초를 열탕에서 가열하여 구절초 추출물을 얻는 단계; (c) 상기 구절초 추출물을 여과하여 상등액을 얻고 농축하는 단계; 및 (d) 상기 구절초 농축액을 동결건조하여 엑기스 분말을 얻는 단계를 포함하는 알레르기 유발물질의 분비를 차단하여 항알레르기 효과를 나타내는 구절초 추출물의 제조방법을 제공한다.The present invention also comprises the steps of (a) washing Gujeolcho with distilled water and pulverizing with a grinder; (b) heating the pulverized gujeolcho in boiling water to obtain gujeolcho extract; (c) filtering the gujeolcho extract to obtain a supernatant and concentrating it; And (d) freeze-drying the gujeolcho concentrate to obtain an extract powder, thereby providing a method of producing gujeolcho extract exhibiting an antiallergic effect by blocking the secretion of allergens.
본 발명에 있어서, 상기 단계 (a)에서 물은 구절초 원료 100g 당 1ℓ를 첨가하는 것을 특징으로 할 수 있고, 상기 단계 (b)에서 구절초를 100~120℃의 열탕에서 2~3시간 동안 가열하여 추출하는 것을 특징으로 할 수 있다.In the present invention, the water in the step (a) may be characterized in that 1 liter per 100 g of gujeolcho raw material is added, and in the step (b) by heating the gujeolcho in a boiling water of 100 ~ 120 ℃ for 2 to 3 hours It may be characterized in that the extraction.
본 발명은 또한, (a) 구절초를 저급 알코올에 침지시켜 초음파 추출기로 추출하여 알콜 추출물을 얻는 단계; (b) 상기 알코올 추출물의 상층액을 여과한 다음, 농축하는 단계; 및 (c) 상기 구절초 농축액을 동결건조하여 엑기스 분말을 얻는 단계를 포함하는 알레르기 유발물질의 분비를 차단하여 항알레르기 효과를 나타내는 구절초 추출물의 제조방법을 제공한다.The present invention also comprises the steps of (a) immersing Gujeolcho in lower alcohol to extract with an ultrasonic extractor to obtain an alcohol extract; (b) filtering the supernatant of the alcohol extract and then concentrating; And (c) freeze-drying the gujeolcho concentrate to obtain an extract powder, thereby providing a method of producing gujeolcho extract exhibiting an antiallergic effect by blocking the secretion of allergens.
본 발명에 있어서, 상기 구절초는 천연 또는 인공 재배한 구절초의 지상부인 것을 특징으로 할 수 있고, 상기 단계 (a)에서 저급 알코올은 95% 에탄올인 것을 특징으로 할 수 있으며, 상기 단계 (a)는 2~4주 추출하는 것을 특징으로 할 수 있다.In the present invention, the gujeolcho may be characterized in that the ground portion of natural or artificially grown Gujeolcho, the lower alcohol in the step (a) may be characterized in that 95% ethanol, the step (a) is It may be characterized by extracting 2 to 4 weeks.
본 발명은 또한, 상기 구절초 추출물을 유효성분으로 함유하는 알레르기성 질환 치료용 조성물을 제공한다.The present invention also provides a composition for treating allergic diseases containing Gujeolcho extract as an active ingredient.
본 발명은 또한, 상기 구절초 추출물을 유효성분으로 함유하는 기능성 식품을 제공한다.The present invention also provides a functional food containing the Gujeolcho extract as an active ingredient.
이하 본 발명을 자세히 설명한다. Hereinafter, the present invention will be described in detail.
본 발명은 한약재의 일종으로서 구절초의 지상부를 유기용매를 이용하여 추출한 구절초 추출물이다. 한의학에서 구절초의 귀경(歸經)은 심(心), 비(脾), 위(胃)이어서 실용할 때의 효능의 방향과 일치한다. 즉, 온중(溫中), 조경(調經), 소화(消化)의 약효가 있어 자궁냉증, 성주기불순, 불임증, 위냉, 소화불량에 사용되고 있다. 주치(主治)에 있어서 여성질환에 널리 쓰이는 것으로 되어 있으나 과학적인 추구가 뒤따라야 할 것이다. 본 발명에서는 이러한 용도이외에도 항알레르기 효과를 규명하였다. The present invention is a kind of herbal medicine Gujeolcho extract extracted from the ground portion of Gujeolcho using an organic solvent. In oriental medicine, Gujeolcho's ear (歸 經) is the heart (heart), rain (위), stomach (胃), which is consistent with the direction of efficacy when practical. In other words, it has the effect of moderate to moderate, landscaping, and digestion, and is used for cold uterine disease, sexual dysfunction, infertility, gastric cooling, and indigestion. It is widely used in women's diseases in the mainstream, but scientific pursuit should follow. In the present invention, the anti-allergic effect was identified in addition to such use.
본 발명자들은 이러한 구절초의 한의학적 약리이론이외에 구절초의 항알레르기 효과를 검색하여 그 추출물이 우수한 약리효과가 있음을 비만세포 및 실험동물 모델에서 확인하였다.The present inventors searched for anti-allergic effects of gujeolcho in addition to the herbal pharmacological theory of gujeolcho and confirmed that the extract has excellent pharmacological effects in mast cells and experimental animal models.
이하, 실시예를 통하여 본 발명을 더욱 상세히 설명하고자 한다. 이들 실시예는 오로지 본 발명을 예시하기 위한 것으로서, 본 발명의 범위가 이들 실시예에 의해 제한되는 것으로 해석되지는 않는 것은 당업계에서 통상의 지식을 가진 자에게 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, it will be apparent to those skilled in the art that the scope of the present invention is not to be construed as being limited by these examples.
실시예 1: 구절초 추출물의 제조Example 1 Preparation of Gujeolcho Extract
본 발명자들은 우수한 항알레르기 효과를 나타내는 구절초 추출물을 얻기 위하여 열탕 및 유기용매 추출방법을 이용하여 구절초 추출물을 제조하였다. The present inventors prepared Gujeolcho extract using boiling water and an organic solvent extraction method in order to obtain Gujeolcho extract showing an excellent antiallergic effect.
(1) 열탕 추출방법(1) boiling water extraction method
구절초의 열탕 추출물을 제조하기 위하여, 구절초를 증류수로 세척하여 분쇄기로 잘게 분쇄한 후 잘게 분쇄된 구절초를 열탕에서 가열하여 구절초 추출물을 얻었다. 이때, 구절초 원료 100g에 대하여 물 1.0ℓ를 첨가한 후, 100~120℃의 열탕에서 2~3시간 동안 가열하여 추출하였고, 일정한 온도를 유지하고 증기로의 손실을 막기 위하여 냉각기를 설치하였다. 이로부터 얻은 구절초 추출물을 여과하여 상등액을 얻고 이를 농축기를 이용하여 농축하였다. 상기 구절초 농축액을 냉동시켜 -80℃에서 동결건조하여 구절초 추출물의 엑기스 분말을 제조하였다. In order to prepare a boiling water extract of gujeolcho, gujeolcho was washed with distilled water, finely pulverized by a grinder, and finely pulverized gujeolcho was heated in a hot water to obtain gujeolcho extract. At this time, 1.0 liter of water was added to 100 g of gujeolcho raw material, and extracted by heating for 2 to 3 hours in a boiling water of 100 ~ 120 ℃, and installed a cooler to maintain a constant temperature and to prevent loss to steam. Gujeolcho extract obtained from this was filtered to obtain a supernatant, which was concentrated using a concentrator. The gujeolcho concentrate was frozen and lyophilized at -80 ° C to prepare extract powder of gujeolcho extract.
(2) 유기용매 추출방법(2) Organic Solvent Extraction Method
구절초의 유기용매 추출물을 제조하기 위하여, 구절초를 저급 알코올에 침지시켜 초음파 추출기로 2~3주 동안 추출하여 알코올 추출물을 얻은 후 상층액만을 분리하여 이를 거즈에 걸러 이물질을 제거하였다. 알코올로는 95% 에탄올을 사용하였다. 상기공정에 의해 얻은 알코올 추출물을 농축기를 이용하여 농축한 후 냉동시키고 -80℃에서 동결건조하여 구절초 추출물의 엑기스 분말을 제조하였다.In order to prepare the organic solvent extract of Gujeolcho, Gujeolcho was immersed in a lower alcohol and extracted for 2 to 3 weeks with an ultrasonic extractor to obtain an alcohol extract. Then, only the supernatant was separated, and the foreign material was removed by gauze. 95% ethanol was used as the alcohol. The alcohol extract obtained by the above process was concentrated using a concentrator, then frozen and lyophilized at -80 ° C to prepare extract powder of gujeolcho extract.
실시예 2: 구절초 추출물의 비만세포에 대한 알레르기 유발물질 분비 억제효과Example 2: Inhibitory Effect of Gujeolcho Extract on Allergen-induced Secretion of Mast Cells
본 발명자들은 상기 실시예 1로부터 추출한 구절초 추출물이 체내에서 알레르기를 유발하는 비만세포로부터 알레르기 유발물질의 분비 억제효과를 조사하였다. RBL 2H3 세포는 글루타민, 항생제와 10% 우혈청이 보충된 최소배지에서 배양되었다. 분비실험을 하기 위하여 세포를 트립신에 의해 수거한 후, 24-well 배양기에 well당 200,000개의 세포를 25ng/ml DNP-특이성 IgE와 같이 배양하였다. 상기 배양된 세포를 PIPES 완충액(25mM PIPES, pH 7.2, 159mM NaCl, 5mM KCl, 0.4mM MgCl2, 1mM CaCl2, 5.6mM glucose, 및 0.1% BSA)으로 세척한 다음, 항원을 첨가하기 전에 30분 동안 전배양 하였다. 전 배양 후 항원을 최종농도 25ng/ml로 첨가하여 자극을 유도하였다. 알레르기 유도물질의 분비정도는 배지중에 분비된 탈과립의 표식자인 hexosaminidase의 활성을 p-nitrophenyl-acetyl-β-D-glucosaminide로 부터 유리된 p-nitrophenol의 양으로 결정하였다.The present inventors investigated the effect of inhibiting the secretion of allergens from gujeolcho extract extracted from Example 1 from mast cells causing allergy in the body. RBL 2H3 cells were cultured in minimal medium supplemented with glutamine, antibiotics and 10% bovine serum. Cells were harvested by trypsin for secretion, and then 200,000 cells per well were incubated with 25ng / ml DNP-specific IgE in a 24-well incubator. The cultured cells were washed with PIPES buffer (25 mM PIPES, pH 7.2, 159 mM NaCl, 5 mM KCl, 0.4 mM MgCl 2 , 1 mM CaCl 2 , 5.6 mM glucose, and 0.1% BSA), and then 30 minutes before the antigen was added. Was precultured. After preculture, the antigen was added at a final concentration of 25 ng / ml to induce stimulation. The secretion of allergens was determined by the amount of p-nitrophenol liberated from p-nitrophenyl-acetyl-β-D-glucosaminide, a marker of degranulation.
(1) 항원에 의한 비만세포로부터 알레르기 유발물질의 분비에 대한 구절초의 억제효과(1) Inhibitory effect of gujeolcho on secretion of allergens from mast cells by antigen
도 1은 알레르기를 유발하는 비만세포 RBL 2H3(Funaba, M. et al., Cell Biol. Int., 27:879-85, 2003; Jeong, H.J. et al., Cytokine, 18:252-9, 2002)에 대한 본 발명에 따른 구절초 추출물의 농도 의존적 억제효과를 나타낸 것으로, 본 발명에 따른 구절초 추출물이 비만세포의 다양한 알레르기 유발물질의 분비를 농도 의존적으로 억제 하였다. 그 최고 분비의 50%를 억제하는 구절초 추출물의 농도(IC50)는 약 49㎍/ml로 낮은 농도에서 높은 억제효과를 나타냈으며, 100㎍/ml에서는 분비가 90%이상 억제되었다.1 shows allergy-inducing mast cell RBL 2H3 (Funaba, M. et al., Cell Biol. Int., 27: 879-85, 2003; Jeong, HJ et al., Cytokine, 18: 252-9, 2002 It showed a concentration-dependent inhibitory effect of the gujeolcho extract according to the present invention for), the gujeolcho extract according to the present invention concentration-dependently inhibited the secretion of various allergens of mast cells. The concentration of Gujeolcho extract (IC 50 ), which inhibits 50% of the highest secretion, was about 49 µg / ml, showing a high inhibitory effect at low concentrations.
(2) Compound 48/80에 의한 비만세포로부터 알레르기 유발물질의 분비에 대한 구절초의 억제효과(2) Inhibitory Effect of Gujeolcho on Secretion of Allergens from Mast Cells by Compound 48/80
도 2는 비만세포 RBL 2H3의 알레르기 유발물질을 다른 물질(compound 48/80)로 자극할 때 본 발명에 따른 추출물의 억제효과를 나타낸 것이다. 도 2에 나타난 바와 같이, compound 48/80에 의한 비만세포로부터 알레르기 유발물질의 분비 역시 구절초 추출물의 의해 저농도(IC50: 약 47.5 ㎍/ml)에서 유의하게 억제되었으며, 100㎍/ml에서는 분비가 90%이상 억제 되었다.Figure 2 shows the inhibitory effect of the extract according to the present invention when stimulating allergens of mast cells RBL 2H3 with other substances (compound 48/80). As shown in FIG. 2, the secretion of allergens from mast cells by compound 48/80 was also significantly inhibited at low concentrations (IC50: about 47.5 μg / ml) by gujeolcho extract. More than% was suppressed.
이로부터 본 발명의 구절초 추출물은 비만세포의 다양한 분비경로를 모두 차단함으로서 알레르기 억제효과를 나타내는 것으로 확인할 수 있었다.From this, gujeolcho extract of the present invention was confirmed to exhibit an allergic inhibitory effect by blocking all the various secretion pathways of mast cells.
실시예 3: 알레르기 동물 모델에서 항알레르기 효과Example 3: Antiallergic Effect in Allergic Animal Models
전신성 알레르기 반응성 아나필락틱 쇼크 동물모델은 compound 48/80을 마우스에 kg당 8mg을 복강내 투여함으로서 얻을 수 있었다. Compound 48/80 투여 한시간 전에 구절초 추출물을 투여하여 아나필락틱 쇼크 억제 효과를 확인 하였다. 표 2에 나타난 바와 같이, 구절초 추출물은 그 반응이 억제 되었으며 kg 당 100mg 투여군에서 35.0%가 억제되었고, kg당 300mg 투여군에서는 48.7%가 억제되었다. 이로부터 본 발명의 구절초 추출물이 시험관에서 뿐 아니라 질병동물 모델에서도 높은 알레르기 질환을 억제하는 효과를 나타내는 것을 확인할 수 있었다. Systemic allergic reactive anaphylactic shock animal models were obtained by intraperitoneally administering compound 48/80 to mice at 8 mg / kg. Gujeolcho was administered an hour before the compound 48/80 administration to confirm the effect of anaphylactic shock. As shown in Table 2, gujeolcho extract was inhibited the reaction, 35.0% was suppressed in the 100mg per kg group, 48.7% in the 300mg per kg group. From this, it was confirmed that gujeolcho extract of the present invention exhibits the effect of suppressing high allergic diseases not only in vitro but also in disease animal models.
이상으로 본 발명 내용의 특정한 부분을 상세히 기술하였는 바, 당업계의 통상의 지식을 가진 자에게 있어서, 이러한 구체적 기술은 단지 바람직한 실시양태일 뿐이며, 이에 의해 본 발명의 범위가 제한되는 것이 아닌 점은 명백할 것이다. 따라서 본 발명의 실질적인 범위는 첨부된 청구항들과 그것들의 등가물에 의하여 정의된다고 할 것이다.Having described the specific part of the present invention in detail, it is obvious to those skilled in the art that such a specific description is only a preferred embodiment, thereby not limiting the scope of the present invention. something to do. Thus, the substantial scope of the present invention will be defined by the appended claims and their equivalents.
상기에서 살펴본 바와 같이, 본 발명에 따른 구절초 추출물은 알레르기를 유발하는 비만세포의 분비작용 및 알레르기의 동물모델에서 알레르기의 유발을 저농도에서 억제하였으며, 한의약에서 사용되어져온 약재임을 고려할 때, 알레르기 발병을 억제하는 기능성 식품이나 의약품 개발에 매우 유용하다.As described above, Gujeolcho extract according to the present invention suppressed the allergen-induced mast cell secretion and allergic induction in the animal model of allergy, considering that it is a drug that has been used in Chinese medicine, allergic development It is very useful for developing functional foods or pharmaceuticals to suppress.
또한, 기존의 증상회복 위주의 치료와는 달리 비만세포에서의 알레르기 유발 물질의 분비를 원천적으로 차단함으로써 다양한 부작용을 제거할 수 있는 의약품이나 기능성 식품으로도 유용하게 사용될 수 있을 것이 기대된다In addition, unlike the existing symptomatic recovery treatment, it is expected to be useful as a medicine or functional food that can eliminate various side effects by blocking the secretion of allergens in mast cells.
도 1은 알레르기를 유발하는 비만세포(RBL 2H3)에 대한 본 발명에 따른 구절초 추출물의 농도 의존적 억제효과를 나타낸 것이다.Figure 1 shows the concentration-dependent inhibitory effect of Gujeolcho extract according to the present invention on allergen-induced mast cells (RBL 2H3).
도 2는 비만세포(RBL 2H3)의 알레르기 유발물질을 다른 물질로 자극할 경우 본 발명에 따른 구절초 추출물의 억제효과를 나타낸 것이다.Figure 2 shows the inhibitory effect of Gujeolcho extract according to the present invention when allergens of mast cells (RBL 2H3) are stimulated with other substances.
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