KR100554079B1 - Lonicera japonica thunb - Google Patents
Lonicera japonica thunb Download PDFInfo
- Publication number
- KR100554079B1 KR100554079B1 KR1020020057527A KR20020057527A KR100554079B1 KR 100554079 B1 KR100554079 B1 KR 100554079B1 KR 1020020057527 A KR1020020057527 A KR 1020020057527A KR 20020057527 A KR20020057527 A KR 20020057527A KR 100554079 B1 KR100554079 B1 KR 100554079B1
- Authority
- KR
- South Korea
- Prior art keywords
- pain
- gold
- silver
- test
- effect
- Prior art date
Links
Images
Classifications
-
- 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)
- A61K36/35—Caprifoliaceae (Honeysuckle family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P25/00—Drugs for disorders of the nervous system
Landscapes
- Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Botany (AREA)
- Neurology (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Biotechnology (AREA)
- Alternative & Traditional Medicine (AREA)
- Biomedical Technology (AREA)
- Mycology (AREA)
- Neurosurgery (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
본 발명은, 행동학적 지표인 신경병리성 통증(neuropathic pain) 모델과 포르말린 테스트(formalin test))를 이용한 동물모델을 이용한 통증에 대한 금은화의 진통효과 검증방법 및 이 진통효과 검증방법을 이용하여 진통효과가 검증된 금은화를 유효성분으로 하는 금은화 진통 제제에 대한 것으로, 신경손상으로 통증을 느끼는 동통환자뿐만 아니라, 일반 동통질환에도 상기 약재의 이용을 통하여 통증 완화효과를 기대할 수 있다.The present invention provides a method for verifying analgesic effect of gold and silver on pain using animal model using neuropathic pain model and formalin test, which are behavioral indicators, and analgesic effect by using this method. For the gold silver analgesic preparations that have been proven as an active ingredient, the pain relief effect can be expected through the use of the medicine not only for pain patients feeling pain due to nerve damage, but also general pain diseases.
Description
도1a 및 도1b는 본 발명에 따른 신경병리성 통증 모델의 통증부위 사진 및 개념도.Figures 1a and 1b is a photograph of the pain area and conceptual diagram of a neuropathological pain model according to the present invention.
도2는 본 발명에 따른 금은화의 기계적 이질통 억제에 미치는 효과 그래프.Figure 2 is a graph of the effect on the mechanical allodynia suppression of gold silver according to the present invention.
도3은 본 발명에 따른 금은화의 냉각 이질통 억제에 미치는 효과 그래프.Figure 3 is a graph of the effect on the cooling allodynia suppression of gold silver coins according to the present invention.
도4a는 본 발명에 사용되는 포르말린 테스트 장치의 사진. Figure 4a is a photograph of the formalin test apparatus used in the present invention.
도4b 및 도4c는 포르말린 테스트에 의한 동물의 행동 사진들. 4B and 4C show behavioral photographs of animals by formalin test.
도5는 본 발명에 따른 금은화의 포르말린 테스트에 따른 금은화의 통증의 억제에 미치는 효과 그래프5 is a graph showing the effect on the inhibition of pain of gold silver coins according to the formalin test of gold silver coins according to the present invention
본 발명은 행동학적 지표인 신경병리성 통증(neuropathic pain) 모델과 포르말린 테스트(formalin test))를 이용한 동물모델을 이용한 통증에 대한 금은화의 진통효과 검증방법 및 이 진통효과 검증방법을 이용하여 진통효과가 검증된 금은화를 유효성분으로 하는 금은화 진통 제제에 대한 것이다.The present invention relates to a method for verifying the analgesic effect of gold and silver on pain using an animal model using a neuropathic pain model and a formalin test, which are behavioral indicators, and an analgesic effect using the method for verifying the analgesic effect. It is about a gold coin analgesic formulation which has proven gold silver coin as an active ingredient.
통증, 특히 만성 통증(chronic pain)은 인간에게 몹시 견디기 어려운 것으로, 만성 통증을 가진 환자는 그 아픔 때문에 정상적인 생활을 영위하는데 많은 어려움을 겪게 된다. 이러한 고통으로부터 해방되기 위해 환자는 통증을 호소하며 도움을 청하게 되며, 통증을 제어하고자 하는 노력은 의학계에서 끊임없이 진행되어져 왔다. 한약은 오랜 기간동안 전통한의학에서 사용되어져 통증에 대한 우수한 치료효과를 보여 왔다. 최근 여러 가지 한약재가 통증 억제에 어떠한 영향을 미치며 어떠한 작용기전에 매개되는지를 연구할 필요성이 제기되어 왔다.Pain, especially chronic pain, is extremely difficult for humans, and patients with chronic pain have a lot of difficulty in living a normal life because of the pain. In order to be freed from these pains, the patient complains of pain and calls for help, and efforts to control pain have been ongoing in the medical community. Chinese medicine has been used in traditional Chinese medicine for a long time and has shown excellent treatment effect for pain. Recently, there has been a need to study how various herbal medicines affect pain inhibition and what mechanism of action is mediated.
기존에 한약물을 이용한 진통억제효과에 대한 보고를 살펴보면, 우슬약침액은 CFA(complete Freund's adjuvant) 관절염 유발 쥐에 작용하여 염증을 억제시키고, 염증상태를 나타내는 백혈구의 총수를 유의성있게 감소시키며, 혈청내 면역 글로불린(Globulin) 및 알부민(albumin)에 작용하여 조직학적으로 근육조직의 괴사를 억제시켜 관절염으로 인한 염증에 대하여 치유 효과를 나타내는 등의 효과가 검증된 바 있었다. 또한, 포공영은 초산법을 이용하여 Writhing syndrome의 빈도를 측정한 결과 포공영의 진통효과가 유의성 있었다. 또한, 초산법과 후지가압법에 의해 갈근을 주재로 한 갈근해기탕의 진통효과가 인정되었다. Previously, the report on the analgesic inhibitory effect using herbal medicines showed that E. coli acts on CFA (complete Freund's adjuvant) arthritis-induced rats to suppress inflammation, significantly reduce the total number of inflammatory cells, and serum It has been proved that the effects on the immunoglobulin (Globulin) and albumin (albumin) to suppress the necrosis of the muscle tissue to show a healing effect on the inflammation caused by arthritis. In addition, pogongyoung showed significant analgesic effect by measuring the frequency of Writhing syndrome using acetic acid method. In addition, the analgesic effect of Galgehaegigi-tang, which is mainly composed of acquaintance, was recognized by acetic acid method and Fuji pressure method.
한편, 이전 연구에서 금은화를 이용하여 항암효과(한두석등, 1998)와 함염효과(최유진등, 2001)가 보고되었으나 여러 통증모델에서 통증억제에 관한 보고는 거의 없었다.On the other hand, anti-cancer effects (Han Doo Seok et al., 1998) and anti-inflammatory effects (Yun Jin Choi, 2001) have been reported using gold and silver coins, but there have been few reports of pain suppression in various pain models.
본 발명은 상기와 같은 종래기술의 필요성과 문제점을 해결하기 위한 것으로, 금은화의 유효한 진통효과를 검증하기 위해 사용되어지는 행동학적 지표인 신경병리성 통증(neuropathic pain) 모델과 포르말린 테스트(formalin test)를 이용한 동물모델을 제공하는 것을 그 목적으로 한다.The present invention is to solve the necessity and problems of the prior art as described above, the neuropathic pain model and formalin test which is a behavioral indicator used to verify the effective analgesic effect of gold and silver coins The object of the present invention is to provide a used animal model.
또한, 본 발명의 목적은, 행동학적 지표인 신경병리성 통증(neuropathic pain) 모델과 포르말린 테스트(formalin test)를 이용한 동물모델을 이용하여 금은화의 유효한 진통효과를 검증하는 것이다.It is also an object of the present invention to verify the effective analgesic effect of gold and silver coins using animal models using neuropathic pain model and formalin test, which are behavioral indicators.
이러한 목적을 달성하기 위하여, 본 발명은, 실험동물의 경골신경과 비복신경의 결찰 및 절단을 통해 신경병리성 동통증후군을 유발할 수 있는 동물모델을 만든 다음, 이질통 검사를 실시하여 통증에 대한 금은화의 진통효과를 검증하는 금은화의 진통효과 검증방법을 제공한다. In order to achieve this object, the present invention, by making the animal model that can cause neuropathic pain syndrome through ligation and cutting of the tibia and non-abdominal nerve of the experimental animal, and then allodynia test for pain relief of gold and silver coins It provides a method to verify the analgesic effect of gold and silver coins.
상기 이질통 검사는 폰 프레이 헤어(von-Frey hair)에 의한 기계적 이질통 검사와 아세톤(acetone)에 의한 냉각 이질통 검사 중 어느 하나일 수 있다.The allodynia test may be any one of a mechanical allodynia test by von-Frey hair and a cold allodynia test by acetone.
상기 신경병리성 동통은 관절염, 근육통, 만성통 중 어느 하나일 수 있다.The neuropathic pain may be any one of arthritis, myalgia, and chronic pain.
또한 본 발명은, 실험동물에 포르말린을 투여하여 실험동물에 직접적 자극을 가한 다음, 금은화의 통증에 대한 진통효과를 검증하는 금은화의 진통효과 검증방 법을 제공한다.In another aspect, the present invention provides a method for verifying the analgesic effect of gold and silver coins to verify the analgesic effect on the pain of gold and silver, and then directly stimulating the experimental animal by administering formalin.
상기 통증은, 염증성 통증(inflammation pain)과 같은 지속성 통증(tonic pain)일 수 있다.The pain may be tonic pain, such as inflammation pain.
또한 본 발명은, 상기 진통효과 검증방법을 이용하여 진통효과가 검증된 금은화의 주요성분을 유효성분으로 하는 진통 제제를 제공한다.In another aspect, the present invention provides an analgesic formulation comprising the main component of the gold and silver coins whose analgesic effect is verified using the analgesic effect verification method.
상기 금은화의 주요성분은, 비닐산(Vanillic acid), 유지놀(Eugenol), 클로로겐산(Chlorogenic acid), 2-페닐에탄올(2-Phenylethanol), 벤질알코올(Benzyl alcohol), 카페인산 메틸에스테르(Caffeic acid methyl ester), 콜린(Choline), β-카로틴(β-Carotene)(all-trans), 크리프토산틴(Cryptoxanthin), 제아크산틴(Zeaxanthin)(all E-form) 중 어느 하나 또는 하나 이상일 수 있다.The main components of the gold and silver coins, vinyl acid (Vanillic acid), eugenol (Eugenol), chlorogenic acid (Chlorogenic acid), 2-phenylethanol (2-Phenylethanol), benzyl alcohol (Benzyl alcohol), caffeic acid methyl ester (Caffeic acid) Methyl ester, Choline, β-Carotene (all-trans), Cryptoxanthin (Cryptoxanthin), Zeaxanthin (all E-form) may be one or more. .
참고로, 본 발명의 한약제인 금은화에 대하여 간단히 살펴보면 다음과 같다.For reference, briefly look at the gold and silver coins of the present invention.
인동초(Lonicera japonica)는 인동과의 반상록 덩굴식물로 산과 들의 양지바른 곳에서 자란다. 한방에서는 인동초의 잎과 줄기를 인동이라 하고, 꽃봉우리를 금은화라 한다. 인동초의 각 마디는 두 송이씨의 꽃을 피우는데, 흰꽃으로 피어났던 꽃은 시간이 지나면서 점차 노란색으로 변하여 인동초의 꽃은 금은화라고 한다.Indonesia (Lonicera japonica) is an evergreen vine plant that grows in sunny areas of mountains and fields. In oriental medicine, the leaves and stems of Indongcho are called indong, and the buds are called gold and silver coins. Each node of Indongcho blooms two seeds. Flowers that bloomed as white flowers gradually turn yellow over time, and the flowers of Indongcho are called gold and silver coins.
실시예1 Example 1
1. 신경병리성 동통 모델 1. Neuropathic Pain Model
가. 개요end. summary
신경병리성 통증 증후군(neuropathic pain syndrome)은, 관절염, 근육통, 만성통과 같이 만성 통증의 하나로서 말초 신경이나 조직이 손상을 받았을 때 발생하게 된다. 신경의 손상으로 인해 유발되는 신경병리성 동통(neuropathic pain)은 매우 고통스러울 뿐만 아니라, 아편제를 비롯한 통상적인 진통제를 사용하더라도 자주 재발된다(Tanelian and Cousins, 1989; Brose and Cousins, 1991). Neuropathic pain syndrome is one of chronic pain, such as arthritis, myalgia, and chronic pain, which occurs when peripheral nerves or tissues are damaged. Neuropathic pain caused by nerve damage is very painful and frequently recurs even with conventional analgesics, including opiates (Tanelian and Cousins, 1989; Brose and Cousins, 1991).
신경병리성 동통의 증후군은 과통증(hyperalgesia), 이질통(allodynia), 자발적 동통(spontaneous pain) 등이 있다. 정상의 경우에도 통증을 유발시킬 수 있는 정도의 자극에 대해 그 이상의 과도한 통증을 유발하는 것을 "과통증"이라 하고, 정상적으로는 통증을 유발시킬 수 없을 정도의 미약한 자극에 민감하게 반응하여 통증을 유발하는 경우를 "이질통"이라 하며, 아무런 자극이 없는 경우에도 통증이 나타나는 것을 "자발적 동통"이라고 한다. Neuropathic pain syndromes include hyperalgesia, allodynia, and spontaneous pain. Even in normal cases, causing excessive pain with respect to the stimulus that can cause pain is called "pain pain", and the pain is sensitively responded to the weak stimulus that cannot normally cause pain. Induced cases are referred to as "allodynia", and pain even when there is no stimulation is called "voluntary pain".
본 발명자들중 이배환 등은 좌골신경의 말초 쪽의 분지를 손상시키는 새로운 모델(Lee et al., 2000)을 개발하였다. 좌골신경에 말초 쪽에서 접근하면 3개의 분지가 있는데, 이들 신경 분지를 여러 가지 집단으로 나누어 선택적으로 손상시켜 신경병리성 동통이 발생되는 정도를 비교한 결과, 총비골신경(common peroneal nerve)은 남겨두고, 경골신경(tibial nerve)과 비복신경(sural nerve)을 손상시킨 집단에서 가장 격심한 동통 증상이 유발되었다(Lee et al., 2000). 이러한 좌골신경 분지의 손상으로 유발된 신경병리성 동통은 손상이 일어난 후 점차 증가하다가 손상 후 2주 정도에서 최고도에 달하였다. 그 후 점차 감소하는 추세를 보였으나 수술 후 약 28주에 이르기까지 지속되었다.Among the present inventors, Lee Bae-hwan et al. Developed a new model (Lee et al., 2000) that damages the peripheral branch of the sciatic nerve. When the sciatic nerve is approached from the peripheral side, there are three branches, and these nerve branches are divided into various groups to selectively damage and compare the degree of neuropathic pain, leaving the common peroneal nerve, The most severe pain symptoms were induced in the tibial and non-sural nerves (Lee et al., 2000). The neuropathic pain caused by the injury of the sciatic nerve branch increased gradually after the injury and reached its peak at about 2 weeks after the injury. Afterwards, the trend gradually decreased, but continued until 28 weeks after surgery.
본 실시예는, 실험동물의 경골신경과 비복신경의 결찰 및 절단을 통해 신경병리성 동통증후군을 유발할 수 있는 동물모델을 만든 다음, 폰프레이 헤어(von-Frey hair)에 의한 기계적 이질통 검사와 아세톤(acetone)에 의한 냉각 이질통 검사를 실시하여 금은화의 처치가 이러한 검사에서 유효한 효과가 나타나는지를 측정하였다. In this embodiment, an animal model that can induce neuropathic pain syndrome through ligation and cleavage of the tibial and gastrocnemius nerves of an experimental animal is made, and then a mechanical allodynia test and acetone using von-Frey hair Cold allodynia testing with acetone was performed to determine whether treatment of gold and silver coins had an effective effect on these tests.
나. 실험내용I. Experiment Content
(1) 신경병리성 동통 모델 제작(1) Model of neuropathic pain
실험동물로는 체중 150-200 gm내외의 흰쥐를 사용한다. Rats weighing 150-200 gm are used as experimental animals.
먼저 할로세인(halothane)으로 쥐를 마취시킨 후 뒷다리 피부의 털을 깎고 포비돈 아이오다인 (povidone iodine) 용액과 이소프로필 알콜(isopropyl alcohol)로 소독한다. 피부를 절개한 후 실체 줌 현미경을 사용하여 좌골신경을 찾아 좌골신경에서 경골 신경(tibial nerve)와 비골신경(common peroneal nerve) 및 비복신경(sural nerve)를 확인한 후 마이크로포셉(microforcep)으로 주변 조직 및 혈관으로 부터 분리시킨 다음, 총비골신경(common peroneal nerve)은 남겨두고, 경골 신경(tibial nerve)와 비복신경(sural nerve)는 현미경 하에서 6.0 명주실(silk thread)로 결찰한 후 미세 수술 가위로 절단한다. 절개된 부위는 카나마이신(kanamycin)을 점적하고 피부를 봉합하여 회복한 후 다음 실험에 들어간다. First, anesthetize the rat with halotane, then shave the hair of the hind limb and disinfect with povidone iodine solution and isopropyl alcohol. After dissection of the skin, the tibial and common peroneal and non-sural nerves are identified from the sciatic nerve by sciatic nerve using a stereoscopic zoom microscope, and then the surrounding tissues with a microforcep. And from the blood vessels, leaving the common peroneal nerve, leaving the tibial and the nasal nerves with a 6.0 silk thread under a microscope, Cut. The incision site is recovered by dripping kanamycin and suturing the skin, and then enters the next experiment.
(2) 한약재의 주입(2) infusion of herbal medicine
금은화 300g을 정확히 평량한 후, 수욕상에서 80% 메탄올을 사용하여 3회 추출하였으며, 추출물을 감압 농축한 후 그 추출물을 동결건조(-66 , 10mmHg)시켜 분말 건조하였다. 건조시켜 얻은 금은화의 수득율은 21.98%이다. After precisely weighing 300 g of gold and silver, it was extracted three times using 80% methanol in a water bath, and the extract was concentrated under reduced pressure, and the extract was lyophilized (-66, 10 mmHg) and powder dried. Yield obtained by drying is 21.98%.
금은화는 생리식염수에 녹여 50mg/10ml/kg, 100mg/10ml/kg, 200mg/10ml/kg, 400mg/10ml/kg의 용량으로 구강내 주입하였다. 각 실험군은 한약재 투여전의 pretest를 실시한 후, 한약재를 투여하고 난 후 2시간 동안 시간경과에 따른 진통효과의 변화를 관찰하였다. 한편, 금은화를 투여하지 않은 실험군을 대조군으로 사용하였다.Gold and silver were dissolved in physiological saline and injected orally at doses of 50 mg / 10 ml / kg, 100 mg / 10 ml / kg, 200 mg / 10 ml / kg, and 400 mg / 10 ml / kg. Each experimental group was subjected to pretest before the administration of medicinal herbs and observed changes in analgesic effect over time for 2 hours after the administration of medicinal herbs. On the other hand, the experimental group was not administered with gold and silver was used as a control.
(3) 동통 반응의 관찰(3) Observation of pain reaction
신경병리성 동통에 관한 행동 검사는 수술전, 수술후 여러 차례에 걸쳐 시행하여 수술 후 2주 까지 관찰한다.Behavioral tests for neuropathic pain are performed several times before and after surgery and observed until 2 weeks after surgery.
외부 자극에 대한 철회반응(withdrawal response)을 보기 위해 실험 동물(쥐)을 망으로 된 우리(cage)에 옮겨 안정시킨 후 실험을 진행한다. In order to see the withdrawal response to external stimuli, the experimental animals (rats) are transferred to a cage of cages and stabilized.
자극은 폰 프레이 필라멘트(von Frey filament), 아세톤(acetone)을 사용하는데, 폰 프레이 필라멘트(von Frey filament)는 기계적 자극에 대한 이질통을 검사하기 위한 것으로서 낮은 강도에서 높은 강도로 자극하여 반응의 역치를 찾는다. 이는 수초 간격으로 좌우 발바닥에 각각 10회씩 자극하여, 발의 철회반응의 횟수를 얻어 백분율로 표시한다. 아세톤(acetone)은 온도자극에 대한 이질통을 검사하기 위한 것으로서, 양쪽 발에 5분 간격으로 각각 5회씩 주사기를 이용, 통각유발부위에 가하여 철회반응의 횟수를 센 다음 백분율을 자료로 삼는다.The stimulus uses von Frey filament, acetone, von Frey filament is used to test allodynia for mechanical stimulation. Find. It stimulates the left and
(4) 실험절차 (4) Experimental procedure
① 신경병리성 통증(neuropathic pain) 수술후 2주일째 되는 날 약물검사를 실시한다.① Neuropathic pain (2 weeks after surgery) The drug test is to be carried out.
② 실험동물을 여러 집단으로 나누고 약물은 구강에 주입한다. 대조군은 생리식염수 동일 용량을 주입한다. ② Divide the test animal into several groups and inject the drug into the oral cavity. The control group is injected with the same dose of physiological saline.
③ 기계적 이질통(mechanical allodynia), 냉각 이질통(cold allodynia)를 검사한다.③ Inspect mechanical allodynia and cold allodynia.
다. 실험결과All. Experiment result
(1) 기계적 이질통에 대한 한약재의 효능(1) Efficacy of Herbal Medicine on Mechanical Allodynia
도2는, 금은화의 기계적 이질통 억제에 미치는 효과 그래프이다.2 is a graph of the effect on the inhibition of mechanical allodynia of gold and silver coins.
도2를 참조하면, 폰 프레이 필라멘트(von frey filament) 자극에 대한 기계적 이질통에 대한 한약재의 진통효과 검사에서 집단간 유의미한 차이를 보였다. 즉, LSD반복측정측정 결과, 60분에서 금은화 400mg/㎏집단과 통제군 사이에 유의미한 차이를 보여(p<0.05) 대조군보다 우수한 이질통 억제효과가 나타났다.Referring to FIG. 2, the analgesic effect test of the herbal medicine for mechanical allodynia on von frey filament stimulation showed a significant difference among the groups. In other words, the LSD repeat measurement result showed a significant difference between the control group and the 400 mg / kg group of gold and silver coins at 60 minutes (p <0.05).
특히, 금은화 50mg/㎏ 집단과 금은화 100mg/㎏ 집단, 200mg/㎏ 집단은 대조 군(controll)과 큰 차이를 보이지 않았으나, 금은화 400mg/㎏집단은 다른 집단들과 60분에서 유의미한 차이를 나타내었다. 따라서, 기계적 이질통 억제를 위해서 실험동물(쥐)의 경우, 금은화를 400mg/㎏ 이상 투여하여야 효과가 있다는 것을 알 수 있었다.In particular, the 50 mg / kg group of gold and silver coins, 100 mg / kg group and 200 mg / kg group showed no significant difference from the control group, but the 400 mg / kg group of gold and silver coins showed a significant difference in 60 minutes from the other groups. Therefore, in order to suppress mechanical allodynia, experimental animals (rats) were found to be effective when 400 mg / kg or more of gold and silver coins were administered.
(2) 냉각 이질통에 대한 한약재의 효능(2) Efficacy of Herbal Medicine on Cooling Allodynia
도3은 금은화의 냉각 이질통 억제에 미치는 효과 그래프이다.3 is a graph of the effect on the inhibition of cooling allodynia of gold and silver coins.
도3을 참조하면, 아세톤(acetone) 자극에 대한 냉각 이질통에 대한 한약재의 진통효과 검사에서 집단간 유의미한 차이는 보이지 않았다(F4,21=0.7, p>0.55). Referring to Figure 3, there was no significant difference between the groups in the analgesic effect test of the herbal medicine for cooling allodynia to acetone stimulation (F4, 21 = 0.7, p> 0.55).
즉, 금은화 50mg/㎏ 집단과 금은화 100mg/㎏ 집단, 200mg/㎏ 집단, 금은화 400mg/㎏집단 모두 대조군(controll)과 큰 차이를 보여 냉각 이질통에 금은화를 미량만 투여하더라도 효과가 있음을 알 수 있었다. 또한, 금은화의 냉각 이질통 억제는 투여하자마자 효과를 나타내었으며, 10분이 경과하여 그 효과가 더욱 뚜렸해 졌다.In other words, the 50mg / kg group of gold and silver coin 100mg / kg, 200mg / kg group, and 400mg / kg group of gold and silver coin showed a big difference from the control group, and it was found that even if only a small amount of gold silver was administered to the cooling allodynia, it was effective. . In addition, the cooling allodynia suppression of gold and silver was effective immediately after administration, the effect became more pronounced after 10 minutes.
실시예2Example 2
2. 포르말린 테스트(formalin test) 동물모델2. Formalin test animal model
가.개요A. Overview
Tail-flick test와 hot-plate test와 같은 전통적인 통증 검사는 대부분 강한 강도의 자극을 일시적으로 제시한다. 이와 같은 방식으로 유발되는 통증 경험 은 지속기간이 짧고, 따라서 자극 자체에 의해 개시되는 조절 기전을 평가하는 데에는 적절하지 못하다. 이에 비해 지속성 통증(tonic pain)은 일시적인 자극에 의해 유발되는 통증과는 다르게 조절될 수 있다. 포르말린 테스트(formalin test)는 일시적으로 제시되는 기계적 또는 온도 자극을 사용한 검사보다도 임상적인 통증을 검사할 수 있는 보다 타당한 모델이라 할 수 있다(Dubuisson & Dennis, 1977; Abbott et al., 1981, 1982; Alreja et al., 1984). Traditional pain tests, such as the tail-flick test and the hot-plate test, often present strong intensity stimuli temporarily. Pain experiences triggered in this way are of short duration and therefore not appropriate for assessing the regulatory mechanism initiated by the stimulus itself. In contrast, tonic pain may be controlled differently than pain caused by transient stimuli. The formalin test may be a more valid model for examining clinical pain than testing with transiently presented mechanical or temperature stimuli (Dubuisson & Dennis, 1977; Abbott et al., 1981, 1982; Alreja et al., 1984).
포르말린(formalin)은 일찍이 통각 유발물질로 사용되어 왔으며(Lewis & Kellgren, 1939; Melzack & Melinkoff, 1974), 쥐에서 말초의 염증 반응을 연구하는 자극으로도 사용되어 왔다(Selye, 1949; Winter, 1965). 1977년 Dubuisson과 Dennis는 포르말린(formalin)으로 유발되는 행동을 자세히 기술함은 물론, 유발된 통증을 수량화하기 위한 방식을 개발하였으며, 이후 수많은 연구자들이 이 검사를 이용해 왔다.Formalin has long been used as a nociceptor (Lewis & Kellgren, 1939; Melzack & Melinkoff, 1974), and has been used as a stimulus to study peripheral inflammatory responses in mice (Selye, 1949; Winter, 1965). ). In 1977, Dubuisson and Dennis described a formalin-induced behavior, as well as a way to quantify the pain caused, and numerous researchers have since used the test.
포르말린(formalin)을 주입하면 통증 반응을 나타내는 두 가지 독특한 페이즈(phase)가 발생한다. 첫 번째 페이즈(phase)는 주입 직후 발생하는 것으로 약 5분간 지속되고, 두 번째 페이즈(phase)는 주입 후 약 20분후 시작한다(Dubbuisson & Dennis, 1977). 인간에게 포르말린(formalin)을 주입하면 국부적인 화끈거리는 통각, 박동통(poorly localized, burning, throbbing pain)을 야기하며, 그 시간경과(time course)는 동물에게서 나타나는 행동적 변화와 일치한다(Dubbuisson & Dennis, 1977). Hunskaar와 Hole의 연구(1987)에 의하면 첫 번째 페이즈(phase)는 포르말린(formalin)이 직접수용기에 작용하여 급성 통증(acute pain)을 야기하고, 두 번째 페이즈(phase)는 지속성 통증(tonic pain)으로서 염증성 통증을 일으키는 것으로 알려져 있다.Injecting formalin results in two distinct phases of pain response. The first phase occurs immediately after the injection, lasts about 5 minutes, and the second phase begins about 20 minutes after the injection (Dubbuisson & Dennis, 1977). Injecting formalin into humans causes localized localized burning, throbbing pain, and the time course coincides with behavioral changes in animals (Dubbuisson & Dennis, 1977). According to a study by Hunskaar and Hole (1987), the first phase is formalin acting directly on the receptor, causing acute pain, and the second phase is tonic pain. It is known to cause inflammatory pain.
이와 같이 포르말린 테스트(formalin test)는 급성 통증(acute pain)과 지속성 통증(tonic pain)을 동시에 반영하는 검사이기 때문에 어떤 약물이 급성 통증(acute pain)을 감소시키는지 아니면 지속성 통증(tonic pain)을 감소시키는가를 알아보는 데에는 매우 적절한 검사법이라 할 수 있다. As such, the formalin test is a test that reflects both acute pain and tonic pain at the same time, which drug reduces acute pain or tonic pain. It's a very good test to see if you can reduce it.
예를 들면, Viana 등(2000)은 레몬그라스(Cymbopogon citratus)에서 추출한 정류(essential oil)가 포르말린 테스트(formalin test)에서 두 번째 페이즈(phase)를 선택적으로 억제함을 보고하였으며, Leal 등(2000)은 쿠마린(coumarin)을 함유하고 있는 브라질산 여러 약초들이 마찬가지로 포르말린 테스트(formalin test)의 두 번째 페이즈(phase)를 억제함을 보고하였다. 또한 Abdel-Fattah 등(2000)은 니겔라 오일(Nigella sativa oil)을 쥐의 구강으로 주입한 경우 포르말린 테스트(formalin test)에서 첫 번째 페이즈(phase)를 억제하였으며, 니겔라 오일(Nigella sativa oil)의 주성분인 티모퀴논(thymoquinone)을 구강, 복강, 뇌실을 통해 주입하였을 때에는 포르말린 테스트(formalin test)에서 첫 번째 뿐만 아니라 두 번째 페이즈(phase)도 억제됨을 보고하였다.For example, Viana et al. (2000) reported that essential oils extracted from lemongrass (Cymbopogon citratus) selectively inhibit the second phase in the formalin test, and Leal et al. (2000) ) Reported that several Brazilian herbs containing coumarin likewise inhibited the second phase of the formalin test. Abdel-Fattah et al. (2000) also inhibited the first phase in the formalin test when Nigella sativa oil was injected into the oral cavity of rats, and Nigella sativa oil. When injection of thymoquinone, the main component of, through the oral cavity, abdominal cavity, and ventricles, it was reported that the formalin test inhibited not only the first phase but also the second phase.
나. 실험내용I. Experiment Content
(1) 실험동물(1) experimental animals
체중 25-35g되는 ICR 마우스를 각 집단에 8-9마리씩 사용하였다. 동물은 한 우리에 4-5마리씩 집단으로 사육하였으며 먹이와 물은 자유로이 섭취하게 하였으며, 통증 검사 당일 최소한 2시간 전에 실험실로 데려와 검사 환경에 적응할 수 있게 하였다.ICR mice weighing 25-35 g were used in each group 8-9. Animals were housed in groups of four to five animals, free to feed and water, and taken to the laboratory at least two hours before the day of pain testing to allow for adaptation to the test environment.
(2) 한약재의 주입(2) infusion of herbal medicine
금은화는 생리식염수에 녹여 50mg/10ml/kg, 100mg/10ml/kg, 200mg/10ml/kg, 400mg/10ml/kg의 용량으로 통증 검사 30분전에 복강내 주입하였다. 약물을 쥐에게 주입하고 포르말린 테스트(formalin test)를 실시하였으며, 이를 정상 쥐(controll)를 사용한 포르말린 테스트(formalin test) 결과와 비교하였다.Gold and silver were dissolved in physiological saline and injected intraperitoneally 30 minutes before the pain test at doses of 50 mg / 10 ml / kg, 100 mg / 10 ml / kg, 200 mg / 10 ml / kg and 400 mg / 10 ml / kg. Drugs were injected into the mice and a formalin test was performed, which was compared with the formalin test results using normal mice.
(3) 동통 반응의 실험장치(3) Experimental apparatus of pain reaction
포르말린(formalin)으로 유발된 통증 반응을 관찰하기 위해 도4a와 같이 투명 아크릴을 이용하여 관찰 상자를 제작하였는데, 그 제원은 12×30×13cm이었다. 관찰 상자의 아래에 도4b와 같이 평면 반사경을 45도의 각도로 설치하고, camcorder(JVC S-VHS)를 이용하여 기록하였다.In order to observe the formalin-induced pain response, an observation box was manufactured using transparent acrylic as shown in FIG. 4A. The specification was 12 × 30 × 13 cm. A flat reflector was installed at an angle of 45 degrees below the observation box as shown in FIG. 4B, and recorded using a camcorder (JVC S-VHS).
(4) 실험절차(4) Experimental procedure
26-gauge needle이 달린 미세주사기(microsyringe)를 사용하여 쥐(mouse) 뒷발등 피하에 2% 포르말린(formalin)을 20㎕ 주사하였다. 주입 직후 쥐를 관찰 상자에 한 마리씩 넣고 통증 행동을 기록하였다. 이러한 통증 행동의 off-line 분석 을 위해, 실험을 마친 뒤 기록된 통증 행동을 TV 모니터(TV monitor)를 이용하여 한 마리의 마우스 당 두 명의 평가자가 포르말린이 주입된 발이나 다리를 핥거나 깨무는 시간의 양을 5분 단위로 평가하였다. A microsyringe with a 26-gauge needle was used to inject 20 μl of 2% formalin in the subcutaneous back of the mouse. Immediately after injection, mice were placed one by one in the observation box and the pain behavior was recorded. For the off-line analysis of these pain behaviors, the time after completion of the experiment was recorded by the TV monitor, where two evaluators per mouse licked or bited the formal or injected foot or leg. The amount of was evaluated in 5 minute increments.
다. 실험결과All. Experiment result
정상동물에게 포르말린(formalin)을 주입한 결과 특징적인 페이즈1(phase 1)과 페이즈2(phase 2)의 통증 반응이 잘 관찰되었다. 이를 근간으로 한약재의 주입이 통증 반응에 미치는 영향을 비교하였는데 그 결과는 도5에 제시되어 있다. 도5는 포르말린(formalin) 주입 후 시간 경과에 따른 통증 행동의 변화 양상을 보여주고 있다. Injecting formalin into normal animals showed a characteristic pain response of phase 1 and phase 2. Based on this, the effect of the infusion of herbal medicine on the pain response was compared and the results are shown in FIG. 5. Figure 5 shows a change in pain behavior over time after formalin injection.
페이즈1(Phase 1)은 포르말린(formalin) 주입 직후부터 10분까지의 반응을 합한 평균을, 페이즈2(phase 2)는 그 이후에 나타나는 반응의 합을 평균하여 제시한 것이다. 도5에서 볼 수 있는 바와 같이, 페이즈1(phase 1)에서는 금은화가 정상(normal or controll) 집단과 아무런 차이가 없었으나(Kruskal-Wallis One Way Analysis of Variance on Ranks, p>.05), 페이즈2(Phase 2)에서는 Kruskal-Wallis One Way ANOVA on Ranks 검증을 수행한 결과 통계적으로 유의미한 차가 있었다(p<0.001). 또한, Dunnett's test를 사용한 multiple comparison 결과, 금은화 집단은 정상 집단과 통계적으로 유의미한 차이가 있었다(p<.05). Phase 1 is the average of the sum of the reactions from just after formalin injection to 10 minutes, and Phase 2 is the average of the sum of the reactions that occur after that. As can be seen in Fig. 5, in phase 1, gold and silver coins were not different from the normal or controll group (Kruskal-Wallis One Way Analysis of Variance on Ranks, p> .05). In Phase 2, the Kruskal-Wallis One Way ANOVA on Ranks test resulted in a statistically significant difference (p <0.001). In addition, as a result of multiple comparison using Dunnett's test, the group of gold and silver coins was significantly different from the normal group (p <.05).
이러한 결과는 금은화가 페이즈1(phase 1)에는 별다른 효과가 없었으므로 급성 통증(acute pain)을 감소시키는 데에는 효과적이지 않지만, 페이즈2(phase 2)에 서는 통증 억제 효과를 나타내므로 염증성 통증(inflammation pain)과 같은 지속성 통증(tonic pain)을 완화시키는데 효과적일 수 있다는 것을 시사한다.These results indicate that gold and silver coins are not effective in reducing acute pain because they have little effect on phase 1, but inflammation pain because they have a pain-inhibiting effect in phase 2. May be effective in alleviating tonic pain, such as).
3. 기타3. Other
이러한 금은화를 유효성분으로 하는 진통제는, 관절염, 근육통, 만성통과 같은 신경병리성 동통이나, 염증성 통증(inflammation pain)과 같은 지속성 통증(tonic pain)에 효과가 있을 것임은 상기 동물모델들을 통해 검증되었다.Analgesics using these gold and silver coins as an active ingredient have been verified in animal models that will be effective in neuropathic pain such as arthritis, myalgia, chronic pain, and tonic pain such as inflammatory pain.
금은화 주요성분은 바닐산(Vanillic acid), 유지놀(Eugenol), 클로로겐산(Chlorogenic acid), 2-페닐에탄올(2-Phenylethanol), 벤질알코올(Benzyl alcohol), 카페인산 메틸에스테르(Caffeic acid methyl ester), 콜린(Choline), β-카로틴(β-Carotene)(all-trans), 크리프토산틴(Cryptoxanthin), 제아크산틴(Zeaxanthin)(all E-form) 등이다. 따라서, 상기 실시예들에 따른 진통효과는 이들을 주요성분으로 함으로써 가능할 것이다. The main components of the gold and silver coins are vanillic acid, eugenol, chlorogenic acid, 2-phenylethanol, benzyl alcohol, and caffeic acid methyl ester. , Choline, β-Carotene (all-trans), Cryptoxanthin (Cryptoxanthin), Zeaxanthin (all E-form), and the like. Therefore, the analgesic effect according to the above embodiments will be possible by making them the main component.
위 실시예및 실험예들에서는 실험용 동물을 쥐로 하였으나, 다른 동물, 예를 들면 개나 원숭이 등일 수 있다. 따라서, 통증 동물모델도 쥐, 마우스 뿐만 아니라 다른 동물일 수 있다.In the above examples and experimental examples, although the experimental animal is a rat, it may be another animal, for example, a dog or a monkey. Thus, the pain animal model can be a mouse, a mouse as well as other animals.
위 실시예에서 제제화 방법으로 추출물 제조만을 언급하였으나, 통상적으로 약제학적으로 허용되는 부형제와 함께 약제학적으로 허용되는 약학적 제제, 예를 들면 주사제, 액제, 시럽제, 정제, 캡슐제, 약침액 등으로 제제화하여 통증억제 예 방 및 치료를 위한 약학적 제제로 사용될 수 있다.In the above embodiment, only the preparation of extracts was mentioned as a formulation method, but in general, pharmaceutically acceptable pharmaceutical preparations such as injections, solutions, syrups, tablets, capsules, medicinal herbs, etc. together with pharmaceutically acceptable excipients It can be formulated and used as a pharmaceutical agent for the prevention and treatment of pain.
본 발명에 따른 약제학적 조성물은 임상적으로 투여시 경구, 복강, 피하 투여 등의 방법으로 투여할 수 있다. The pharmaceutical composition according to the present invention can be administered by a method such as oral, intraperitoneal, subcutaneous administration when clinically administered.
본 발명에 따른 약제학적 조성물의 임상적 투여용량은 환자의 연령, 증상, 투여제형 또는 약물의 종류에 따라 다양하게 조절할 수 있다. 특정한 상태에서 바람직한 투여량을 결정하는 것은 공지된 기술에 해당한다.The clinical dosage of the pharmaceutical composition according to the present invention can be variously adjusted according to the age, symptoms, dosage form or type of drug of the patient. Determining the desired dosage in a particular condition corresponds to known techniques.
4. 참고문헌4. References
Abbott FV, Franklin KB, Ludwick RJ, and Melzack R. Apparent lack of tolerance in the formalin test suggests different mechanisms for morphine analgesia in different types of pain. Pharmacol Biochem Behav 15:637-640, 1981.Abbott FV, Franklin KB, Ludwick RJ, and Melzack R. Apparent lack of tolerance in the formalin test suggests different mechanisms for morphine analgesia in different types of pain. Pharmacol Biochem Behav 15: 637-640, 1981.
Abbott FV, Melzack R, and Samuel C. Morphine analgesia in tail-flick and formalin pain tests is mediated by different neural systems. Exp Neurol 75:644-651, 1982.Abbott FV, Melzack R, and Samuel C. Morphine analgesia in tail-flick and formalin pain tests is mediated by different neural systems. Exp Neurol 75: 644-651, 1982.
Abdel-Fattah AM, Matsumoto K, and Watanabe H. Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. Eur J Pharmacol 400:89-97, 2000.Abdel-Fattah AM, Matsumoto K, and Watanabe H. Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. Eur J Pharmacol 400: 89-97, 2000.
Alreja V, Mutalik P, Nayar U, and Manchanda SK. The formalin test: a tonic pain model in the primate. Pain 20:97-104, 1984.Alreja V, Mutalik P, Nayar U, and Manchanda SK. The formalin test: a tonic pain model in the primate. Pain 20: 97-104, 1984.
Brose and Cousins. Subcutaneous lidocaine for treatment of neuropathic cancer pain. Pain 45:145-8, 1991.Brose and Cousins. Subcutaneous lidocaine for treatment of neuropathic cancer pain. Pain 45: 145-8, 1991.
Dubuisson D and Dennis SG. The formalin test: a quantitative study of the analgesic effects of morphine, meperidine, and brain stem stimulation in rats and cats. Pain 4:161-174, 1977.Dubuisson D and Dennis SG. The formalin test: a quantitative study of the analgesic effects of morphine, meperidine, and brain stem stimulation in rats and cats. Pain 4: 161-174, 1977.
Hunskaar S and Hole K. The formalin test in mice: dissociation between inflammatory and non-inflammatory pain. Pain 30:103-114, 1987.Hunskaar S and Hole K. The formalin test in mice: dissociation between inflammatory and non-inflammatory pain. Pain 30: 103-114, 1987.
Leal LKAM, Ferreira AAG, Bezerra GA, Matos FJA, and Viana GSB. Antinociceptive, anti-inflammatory and bronchodilator activities of Brazilian medicinal plants containing coumarin: a comparative study. J Ethnopharmacol 70:151-159, 2000.Leal LKAM, Ferreira AAG, Bezerra GA, Matos FJA, and Viana GSB. Antinociceptive, anti-inflammatory and bronchodilator activities of Brazilian medicinal plants containing coumarin: a comparative study. J Ethnopharmacol 70: 151-159, 2000.
Lee, B.H., Baik, E.J., Kim, E.J., Lee, S.H., & Moon, C.H. Development of behavioral signs of neuropathic pain following injury to distal sciatic nerve branches. Neuroscience Abstracts, p.119, 1996a.Lee, B.H., Baik, E.J., Kim, E.J., Lee, S.H., & Moon, C.H. Development of behavioral signs of neuropathic pain following injury to distal sciatic nerve branches. Neuroscience Abstracts, p. 119, 1996a.
Lee, B.H., Baik, E.J., Lee, S.H., & Moon, C.H. Injuries of different distal sciatic nerve branches differentially produce neuropathic pain in rats. Abstracts - 8th World Congress on Pain, p.27, 1996b.Lee, B.H., Baik, E.J., Lee, S.H., & Moon, C.H. Injuries of different distal sciatic nerve branches differentially produce neuropathic pain in rats. Abstracts-8th World Congress on Pain, p. 27, 1996b.
Lee, B.H. Won, R., Baik, E.J., Lee, S.H., & Moon, C.H. An animal model of neuropathic pain employing injury to the sciatic nerve branches. NeuroReport, 11(4):657-661, 2000a.Lee, B.H. Won, R., Baik, E.J., Lee, S.H., & Moon, C.H. An animal model of neuropathic pain employing injury to the sciatic nerve branches. NeuroReports, 11 (4): 657-661, 2000a.
Lews T and Kellgren JH. Observations relating to referred pain, visceromotor reflexes and other associated phenomena. Clin Sci 4:47-71, 1939.Lews T and Kellgren JH. Observations relating to referred pain, visceromotor reflexes and other associated phenomena. Clin Sci 4: 47-71, 1939.
Melzack R and Melinkoff DF. Analgesia produced by brain stimulation: Evidence of a prolonged onset period. Exp Neurol 43:369-374, 1974.Melzack R and Melinkoff DF. Analgesia produced by brain stimulation: Evidence of a prolonged onset period. Exp Neurol 43: 369-374, 1974.
Selye H. Futher studies concerning the participation of the adrenal cortex in the pathogenesis of arthritis. Br Med J, 2:1129-1135, 1949.Selye H. Futher studies concerning the participation of the adrenal cortex in the pathogenesis of arthritis. Br Med J, 2: 1129-1135, 1949.
Tanelian and Cousins, Combined neurogenic and nociceptive pain in a patient with Pancoast tumor managed by epidural hydromorphone and oral carbamazepine. Pain. 1989 Jan;36(1):85-8.Tanelian and Cousins, Combined neurogenic and nociceptive pain in a patient with Pancoast tumor managed by epidural hydromorphone and oral carbamazepine. Pain. 1989 Jan; 36 (1): 85-8.
Viana GSB, Vale TG, Pinho RSN, and Matos FJA. Antinociceptive effect of the essential oil from Cymbopogon citratus in mice. J Ethnopharmacol 70:323-327, 2000.Viana GSB, Vale TG, Pinho RSN, and Matos FJA. Antinociceptive effect of the essential oil from Cymbopogon citratus in mice. J Ethnopharmacol 70: 323-327, 2000.
Winter CA. Anti-inflammatory testing methods: comparative evaluation of indomethacin and other agents. In: S. Gavattini and M.N.G. Dukes (Eds), Non-Steroidal Anti-Inflammatory Drugs, Vol. 82, Excerpa Medica International Congress Series, Amsterdam, 1965, pp. 190-202.Winter CA. Anti-inflammatory testing methods: comparative evaluation of indomethacin and other agents. In: S. Gavattini and M.N.G. Dukes (Eds), Non-Steroidal Anti-Inflammatory Drugs, Vol. 82, Excerpa Medica International Congress Series, Amsterdam, 1965, pp. 190-202.
최유진, 정성숙, 홍석진, 임희순, 손재범. 금은화와 포공영추출물이 첨가된 치약의 치면세균막 및 치은염에 미치는 영향대한구강보건학회지. Vol.25, No.4: 347-58, 2001. Choi Yu-jin, Jung Sung-sook, Hong Seok-jin, Lim Hee-soon, Son Jae-bum. The Effect of Toothpaste with Gold Silver Coated and Pogongyoung Extracts on the Dental Bacterial Membrane and Gingivitis Korean Journal of Oral Health. Vol. 25, No. 4: 347-58, 2001.
한두석, 배경현, 최은규,곽정숙. 한국산 생약으로부터 항암물질의 개발 (제6 보) 금은화 Ethyl Acetate 가용성 분획의 인체 구강유상피암종세포에 미치는 세포독성작용. 생약학회지, Vol.29, No.1,22-28,1998. Doo Seok Han, Background Hyun, Choi Eun Kyu, Kwak Jeong Sook. Development of Anticancer Substances from Korean Herbal Medicines (Chapter 6) Cytotoxic Activity of Ethyl Acetate Soluble Fraction on Human Oral Carcinoid Carcinoma Cells. Korean Journal of Pharmacognosy, Vol. 29, No. 1, 22-28, 1998.
이상과 같이 본 발명에 의하면, 금은화의 유효한 진통효과를 검증하기 위해 사용되어지는 행동학적 지표인 신경병리성 통증(neuropathic pain) 모델과 포르말린 테스트(formalin test))를 이용한 동물모델을 제공하는 효과가 있다. As described above, according to the present invention, there is an effect of providing an animal model using a neuropathic pain model and formalin test, which are behavioral indicators used to verify the effective analgesic effect of gold and silver coins. .
또한, 본 발명은, 행동학적 지표인 신경병리성 통증(neuropathic pain) 모델과 포르말린 테스트(formalin test))를 이용한 동물모델을 이용하여 금은화의 유효한 진통효과를 검증할 수 있는 효과가 있다.In addition, the present invention is effective to verify the effective analgesic effect of gold and silver coins using animal models using the neuropathic pain model (formalin test) and the behavioral indicators.
이와 같이, 본 발명은, 우수한 진통효과를 보일 것으로 추정되는 금은화를 정제하여 추출한 후, 신경병리성 통증(neuropathic pain) 모델에서 기계적 및 냉각 이질통(mechanical and cold allodynia)에 대한 억제효과와 포르말린 테스트(formalin test))에서 직접적 자극에 대한 통증 및 염증성 반응에 의한 통증에 대한 억제 효과를 검사하는 등의 최근 진행되는 통증에 대한 여러 가지 지표에 대한 검사를 실시하여, 금은화의 우수한 진통억제 효과를 발견하였다. As described above, the present invention, after purifying the gold and silver coins which are estimated to have excellent analgesic effect, and the inhibitory effect against mechanical and cold allodynia and formalin test in the neuropathic pain model In the test), various indicators of recent pain, such as examining the effects of pain on direct stimulation and pain by inflammatory response, were tested, and the excellent analgesic suppression effect of gold and silver coins was found.
본 발명은 신경병리성 동통 모델과 포르말린 테스트(formalin test)에서 금은화의 우수한 진통효과를 입증함으로써, 신경손상으로 통증을 느끼는 동통환자뿐만 아니라, 일반 동통질환에도 상기 약재의 이용을 통하여 통증 완화효과를 기대해볼 수 있을 것이다. The present invention demonstrates the excellent analgesic effect of gold and silver coins in the neuropathological pain model and formalin test, and expects the pain relief effect through the use of the medicine not only for pain patients feeling pain due to nerve damage, but also for general pain diseases. You can try.
Claims (8)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020020057527A KR100554079B1 (en) | 2002-09-23 | 2002-09-23 | Lonicera japonica thunb |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020020057527A KR100554079B1 (en) | 2002-09-23 | 2002-09-23 | Lonicera japonica thunb |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| KR20040026170A KR20040026170A (en) | 2004-03-30 |
| KR100554079B1 true KR100554079B1 (en) | 2006-02-22 |
Family
ID=37328680
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020020057527A KR100554079B1 (en) | 2002-09-23 | 2002-09-23 | Lonicera japonica thunb |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR100554079B1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102895147A (en) * | 2012-09-25 | 2013-01-30 | 广州中汉口腔用品有限公司 | Toothpaste with anti-inflammation and anti-allergic effects and making technology thereof |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR101072264B1 (en) | 2008-02-19 | 2011-10-11 | 환인제약 주식회사 | Composition comprising mixed herbal extract of Lonicera japonica THUNB and Anemarrhena asphodeloides BUNGE preventing and treating arthritis |
| WO2009104913A2 (en) * | 2008-02-19 | 2009-08-27 | University-Industry Cooperation Group Of Kyung Hee University | Composition comprising an extract of mixed herbs with lonicera japonica thunb and anemarrhena asphodeloides bunge for preventing and treating arthritic diseases |
| KR101016837B1 (en) * | 2008-02-19 | 2011-02-22 | 환인제약 주식회사 | Composition comprising the extract of Lonicera japonica THUNB. preventing and treating arthritis |
| KR101326241B1 (en) * | 2012-02-06 | 2013-11-11 | 순천향대학교 산학협력단 | Pharmaceutical Composition for improving anti pain Comprising Lonicerae Flos Extracts |
| CN110038023B (en) * | 2019-04-19 | 2021-05-07 | 温州医科大学 | Anti-allergic pharmaceutical composition for replacing cocklebur fruit |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR940001893A (en) * | 1992-07-02 | 1994-02-16 | 강삼식 | Anti-inflammatory, anti-allergic and antirheumatic agents of biflavonoids from Lonicera japonica |
| KR19990073884A (en) * | 1998-03-04 | 1999-10-05 | 강삼식 | Analgesic with biflavonoids and derivatives as active ingredient |
| KR20040023392A (en) * | 2002-09-11 | 2004-03-18 | 에스케이케미칼주식회사 | Extraction and purification method of constituents from stem of Lonicera japonica Thunb. and the constituents of injection drug |
-
2002
- 2002-09-23 KR KR1020020057527A patent/KR100554079B1/en not_active IP Right Cessation
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR940001893A (en) * | 1992-07-02 | 1994-02-16 | 강삼식 | Anti-inflammatory, anti-allergic and antirheumatic agents of biflavonoids from Lonicera japonica |
| KR19990073884A (en) * | 1998-03-04 | 1999-10-05 | 강삼식 | Analgesic with biflavonoids and derivatives as active ingredient |
| KR20040023392A (en) * | 2002-09-11 | 2004-03-18 | 에스케이케미칼주식회사 | Extraction and purification method of constituents from stem of Lonicera japonica Thunb. and the constituents of injection drug |
Non-Patent Citations (3)
| Title |
|---|
| J. Ethnopharmacol. 70(3), 323-327(2000. 06.) * |
| Neurosci. Lett. 291(1), 29-32(2000. 09.) * |
| 약학회지, 43(1), 117-123 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102895147A (en) * | 2012-09-25 | 2013-01-30 | 广州中汉口腔用品有限公司 | Toothpaste with anti-inflammation and anti-allergic effects and making technology thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| KR20040026170A (en) | 2004-03-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Ojewole et al. | Hypoglycaemic and hypotensive effects of Momordica charantia Linn (Cucurbitaceae) whole-plant aqueous extract in rats: Cardiovascular topics | |
| Mate et al. | Evaluation of anti-nociceptive activity of Cissus quadrangularis on albino mice | |
| KR100554079B1 (en) | Lonicera japonica thunb | |
| US10561678B2 (en) | Composition for treating neuropathy, process and method of treatment thereof | |
| KR100554082B1 (en) | Nelumbo nucifera gaertn | |
| US6166085A (en) | Method of producing analgesia | |
| KR101500485B1 (en) | Compositions for Alleviating, Preventing or Treating Pain Comprising Ilex paraguayensis Extracts as Active Ingredients | |
| KR20010103341A (en) | Extract of Uncariae Ramulus having neuroprotective effects and pharmaceutical preparations containing the same | |
| Akram et al. | Antidiabetic Activity of root of Terminalia chebula on alloxan induced diabetic rat | |
| KR101680840B1 (en) | Compositions for Alleviating, Preventing or Treating Pain Comprising Cnidium officinale Makino Extracts as Active Ingredients | |
| KR100341368B1 (en) | Treatment of cardio- vascular disease and Its process | |
| KR100715939B1 (en) | Therapeutic Agent for Narcotics Addiction | |
| Al-Khazraji | The pain decreasing effect of the alcoholic extract of Trachyspermum ammi (L.)(Ajwain) in experimental animals | |
| KR20040060226A (en) | Acanthopanacis Cortex | |
| CN113827587A (en) | Application of salvianolic acid A in preparing medicine for preventing thrombotic cerebral ischemia | |
| KR20020078850A (en) | Extract of Angelicae Gigantis Radix having neuroprotective effects and pharmaceutical composition containing the same | |
| KR20210088775A (en) | Composition for preventing or treating neuropathic pain comprising extracts of herbal mixture | |
| KR101359760B1 (en) | The pharmaceutical composition comprising the extract of Astilbe chinesis for preventing and treating the degenerative brain disease | |
| KR20020087723A (en) | Extract of Rheum palmatum L. having neuroprotective effects and pharmaceutical composition containing the same | |
| Arefin et al. | Analgesic, anti-inflammatory, anticonvulsant and CNS effect of the ethanolic extract of Butea monosperma roots | |
| CA2480223C (en) | A composition for treating gastric ulcer and a process for preparing the same | |
| EP0870499A1 (en) | Oral use of (+)-0-Demethyltramadol as analgesic | |
| Nwinyi et al. | Evaluation of aqueous-methanol stem bark extract of Stereospermum kunthianum Cham.(Family: Bignoniaceae) for Anti-inflammatory and antinociceptive effects | |
| 정기태 | Effect of Sec-O-glucosylhamaudol on Mechanical Allodynia in a Rat Model of Postoperative Pain | |
| US7687081B2 (en) | Herbal formulation useful as local anesthetic |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| A201 | Request for examination | ||
| E902 | Notification of reason for refusal | ||
| E90F | Notification of reason for final refusal | ||
| E701 | Decision to grant or registration of patent right | ||
| GRNT | Written decision to grant | ||
| FPAY | Annual fee payment |
Payment date: 20130215 Year of fee payment: 8 |
|
| FPAY | Annual fee payment |
Payment date: 20140214 Year of fee payment: 9 |
|
| FPAY | Annual fee payment |
Payment date: 20141226 Year of fee payment: 10 |
|
| FPAY | Annual fee payment |
Payment date: 20160121 Year of fee payment: 11 |
|
| FPAY | Annual fee payment |
Payment date: 20180814 Year of fee payment: 13 |
|
| LAPS | Lapse due to unpaid annual fee |