KR20050030652A - PROSTAGLANDIN E.sub.2 INHIBITOR COMPRISING EXTRACT FROM ANGELICA DAHURICA OR FURANOCOUMARIN DERIVATIVE DERIVED FROM THE SAME - Google Patents

PROSTAGLANDIN E.sub.2 INHIBITOR COMPRISING EXTRACT FROM ANGELICA DAHURICA OR FURANOCOUMARIN DERIVATIVE DERIVED FROM THE SAME Download PDF

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KR20050030652A
KR20050030652A KR1020030066584A KR20030066584A KR20050030652A KR 20050030652 A KR20050030652 A KR 20050030652A KR 1020030066584 A KR1020030066584 A KR 1020030066584A KR 20030066584 A KR20030066584 A KR 20030066584A KR 20050030652 A KR20050030652 A KR 20050030652A
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formula
prostaglandin
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임순성
정상훈
신국현
신현경
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학교법인 한림대학교
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/366—Lactones having six-membered rings, e.g. delta-lactones
    • A61K31/37—Coumarins, e.g. psoralen
    • 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)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A61K36/232—Angelica

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Abstract

A prostaglandin E2 inhibitor comprising an extract of Angelica dahurica or furanocoumarin derivatives derived from the same extract is provided, which prostaglandin E2 inhibitor inhibits inflammation response, pain and cancer, so that it can be useful as anti-inflammatory drug, anodynia and anticancer drug. The prostaglandin E2 inhibitor comprises a compound represented by formula (1), wherein R1 is H, OCH3, OCH2-CH=C(CH3)2 or -O-OH-OCH3; R2 is H, OCH2-CH=C(CH3)2 or -O-OH-OH; R3-R4 and R5-R6 are independently HC=CH or H2C-CH2; and the compound of formula (1) is selected from compounds of formula (2) to formula (11). The prostaglandin E2 inhibitor comprises the extract of Angelica dahurica, wherein the Angelica dahurica extract is produced by extracting Angelica dahurica with methanol, hexane, methylene chloride and ethyl acetate. A pharmaceutical composition for prevention and treatment of diseases associated with prostaglandin E2 comprises the prostaglandin E2 inhibitor.

Description

구릿대 추출물 또는 이로부터 유래한 화합물을 포함하는 프로스타글란딘 Ｅ₂ 합성 억제제{PROSTAGLANDIN Ｅ.sub.2 INHIBITOR COMPRISING EXTRACT FROM ANGELICA DAHURICA OR FURANOCOUMARIN DERIVATIVE DERIVED FROM THE SAME}PROSTAGLANDIN E.sub.2 INHIBITOR COMPRISING EXTRACT FROM ANGELICA DAHURICA OR FURANOCOUMARIN DERIVATIVE DERIVED FROM THE SAME}

[발명이 속하는 기술분야][TECHNICAL FIELD OF THE INVENTION]

본 발명은 구릿대 추출물 또는 이로부터 유래한 화합물을 포함하는 프로스타글란딘 E2 합성 억제제에 관한 것으로, 보다 상세하게는 COX-2 및 mPGES 발현을 저해하여 프로스타글란딘 E2 합성을 억제하므로써 소염제 및 진통제로 사용가능한 구릿대 추출물 또는 이로부터 유래한 퓨라노쿠마린계 화합물에 관한 것이다.The present invention relates to a prostaglandin E 2 synthesis inhibitor comprising a copper extract or a compound derived therefrom, and more particularly, a copper bar that can be used as an anti-inflammatory agent and an analgesic agent by inhibiting prostaglandin E 2 synthesis by inhibiting COX-2 and mPGES expression. It relates to an extract or a furanocoumarin compound derived therefrom.

[종래기술][Private Technology]

구릿대(Angelica dahurica)는 약학적으로 매우 중요한 식물중 하나로, 이의 뿌리는 백지라 불리우며 한의약에서 감기, 치통, 신경통 및 출혈로 인한 두통을 완화시키는 약재로 이용되고 있다(Encyclopedia of traditional Chinese materia medica, Shanghai, China: The people's press, 1977; Vol. IV: 4534).Angelica dahurica is one of the most important pharmacological plants. Its root is called white paper and is used as a medicine to relieve headaches caused by colds, toothaches, neuralgia and bleeding in Chinese medicine (Encyclopedia of traditional Chinese materia medica, Shanghai). , China: The people's press, 1977; Vol. IV: 4534).

한편, 활성화된 대식세포는 프로스타글란딘(prostaglandin: PG) E2를 생산하여 염증성 반응동안 고통과 열을 야기한다. 프로스타글란딘 E2는 세포막 인지질로부터 방출된 아라키도닌산(arachidonic acid)으로부터 합성되며, 이때 2가지 효소가 작용한다. 이중 하나인 사이클로옥시게나제(cyclooxygenase: 이하 "COX"라 한다.)는 아라키도닌산을 플로스타글란딘 H2로 전환시키고, 다른 효소인 PGE 합성효소(PGES)는 프로스타글란딘 H2를 프로스타글란딘 E2로 전환시킨다. 상기 두 효소 COX 및 PGES는 두 가지 형태가 있는 것으로 보고되고 있다(Smith WL, Marnett LJ. (1991) Biochimica Biophysica Acta, 1083: 1-17 ; Hsi LC, Hoganson CW, Babcock GT, Smith WL. (1994) Biochemical and Biophysical Research Communications, 202: 1592-8). COX-1와 세포내 PGES(이하 "cPGES"라 한다.)는 일반적으로 발현되는 형태이고(O’Banion MK, Winn VD, Young DA. (1992) Proceedings of the National Academy of Sciences of the United States of America, 89: 4888-92), 반면에 COX-2와 마이크로솜의 PGES(이하 "mPGES"라 한다.)는 여러가지의 전염증 촉진자(proinflammatory stimulators)들에 의하여 발현된다(O’Sullivan MG, Chilton FH, Huggins EM Jr, McCall CE (1992) Journal of Biolological Chemistry, 267: 14547-50; Nishizuka Y. (1992) Science, 258: 607-14 ; Appleby SB, Ristimaki A, Neilson K, Narko K, Hla T. (1994) Biochemical Journal 1994; 302: 723-7). 특히 COX-1은 위장관, 신장 및 혈소판에서 정상적으로 분포하여 점액과 바이카보네이트의 분비 및 점막 혈류를 증가시키며 점막 상피세포이 증식을 도와주며, COX-2는 염증 부위에서 유도된다.Activated macrophages, on the other hand, produce prostaglandin (PG) E 2 , causing pain and fever during the inflammatory response. Prostaglandin E 2 is synthesized from arachidonic acid released from cell membrane phospholipids, with two enzymes acting. One of them, cyclooxygenase (hereinafter referred to as "COX"), converts arachidonic acid to flostaglandin H 2 , and another enzyme, PGE synthase (PGES), converts prostaglandin H 2 to prostaglandin E 2. Switch to The two enzymes COX and PGES are reported to be in two forms (Smith WL, Marnett LJ. (1991) Biochimica Biophysica Acta, 1083: 1-17; Hsi LC, Hoganson CW, Babcock GT, Smith WL. (1994) ) Biochemical and Biophysical Research Communications, 202: 1592-8). COX-1 and intracellular PGES (hereinafter referred to as "cPGES") are commonly expressed forms (O'Banion MK, Winn VD, Young DA. (1992) Proceedings of the National Academy of Sciences of the United States of America, 89: 4888-92), while COX-2 and microsomal PGES (hereinafter referred to as "mPGES") are expressed by a variety of proinflammatory stimulators (O'Sullivan MG, Chilton). FH, Huggins EM Jr, McCall CE (1992) Journal of Biolological Chemistry, 267: 14547-50; Nishizuka Y. (1992) Science, 258: 607-14; Appleby SB, Ristimaki A, Neilson K, Narko K, Hla T (1994) Biochemical Journal 1994; 302: 723-7. In particular, COX-1 is normally distributed in the gastrointestinal tract, kidney and platelets to increase the secretion of mucus and bicarbonate and increase the mucosal blood flow, and help the mucosal epithelial cell proliferation, COX-2 is induced at the site of inflammation.

흔히 사용되는 비스테로이드성 소염제(NSAIDs)는 아라키도닌산으로부터 프로스타글란딘의 생성을 억제하는 작용을 하는 것으로, 대표적인 것으로 셀레코십(celecoxib) 로펙토십(rofecoxib), 멜로시캄(meloxicam), 디클로페낙(diclofenac), 이부프로펜(ibuprofen), 피록시캄(piroxicam), 아스피린(aspirin) 및 인도메타신(indomethacin)이 있다. 그러나 비스테로이성 소염제의 일부 약물은 COX-1과 COX-2를 비선택적으로 억제하거나 또는 COX-1에 약간의 선택성을 갖고 있어, 염증반응 이외의 정상적인 프로스타글란딘의 합성을 저해하여 위장궤양, 위장의 불내성, 혈소판 응집 차단 및 자궁 운동성 억제와 같은 부작용을 발생시킨다. 따라서 근래에는 COX-1에는 거의 영향을 미치지 않으면서 COX-2를 선택적으로 저해할 수 있는 소염제 개발에 관심이 집중되고 있다.Commonly used nonsteroidal anti-inflammatory drugs (NSAIDs) act to inhibit the production of prostaglandins from arachidonic acid. , Ibuprofen, pyroxicam, aspirin and indomethacin. However, some drugs of non-steroidal anti-inflammatory drugs have non-selective inhibition of COX-1 and COX-2 or some selectivity to COX-1, which inhibits the synthesis of normal prostaglandins other than inflammatory reactions. Side effects such as intolerance, blocking platelet aggregation and suppressing uterine motility. Therefore, attention has recently been focused on the development of anti-inflammatory agents that can selectively inhibit COX-2 with little effect on COX-1.

따라서, 본 발명은 식물로부터 프로스타글란딘 E2 합성 억제효과를 갖는 식물 추출물 또는 식물 유래 화합물을 제공하는 것을 목적으로 한다.Therefore, an object of the present invention is to provide a plant extract or a plant-derived compound having a prostaglandin E 2 synthesis inhibitory effect from a plant.

또한 본 발명은 COX-2에 선택적 저해성을 가지는 약학적 조성물을 제공하는 것을 목적으로 한다. It is another object of the present invention to provide a pharmaceutical composition having selective inhibitory effect on COX-2.

상기 목적을 달성하기 위하여 본 발명은 하기 화학식 1로 표시되는 화합물을 유효성분으로 포함하는 프로스타글란딘 E2 합성 억제제를 제공한다:In order to achieve the above object, the present invention provides a prostaglandin E 2 synthesis inhibitor comprising a compound represented by the following formula (1) as an active ingredient:

(화학식 1)(Formula 1)

상기 화학식 1에서,In Chemical Formula 1,

R1은 H, OCH3, OCH2-CH=C(CH3)2 또는 이고;R 1 is H, OCH 3 , OCH 2 —CH═C (CH 3 ) 2 or ego;

R2는 H, OCH2-CH=C(CH3)2 또는 이고; 및R 2 is H, OCH 2 —CH═C (CH 3 ) 2 or ego; And

R3-R4 및 R5-R6은 각각 독립적으로 HC=CH 또는 H2 C-CH2 이다.R 3 -R 4 and R 5 -R 6 are each independently HC═CH or H 2 C—CH 2 .

또한 본 발명은 구릿대 추출물을 유효성분으로 포함하는 프로스타글란딘 E2 합성 억제제를 제공한다.In another aspect, the present invention provides a prostaglandin E 2 synthesis inhibitor comprising a copper leaf extract as an active ingredient.

또한 본 발명은 프로스타글란딘 E2 합성 억제제를 포함하는 프로스타글란딘 E2 관련 질환 치료 및 예방용 약학적 조성물을 제공한다.The present invention also provides a pharmaceutical composition for the treatment and prevention of prostaglandin E 2 related diseases comprising a prostaglandin E 2 synthesis inhibitor.

이하 본 발명을 상세히 설명한다.Hereinafter, the present invention will be described in detail.

본 발명은 프로스타글란딘 E2 합성 억제활성을 갖는 구릿대 추출물 및 상기 추출물에서 유래한 상기 화학식 1로 표시되는 화합물에 관한 것이다.The present invention relates to a copper leaf extract having a prostaglandin E 2 synthesis inhibitory activity and a compound represented by the formula (1) derived from the extract.

본 발명의 구릿대 추출물은 구릿대를 물 또는 유기용매로 추출하여 제공된다. 바람직하기로는 탄소수 1 내지 5의 알콜, 헥산, 메틸렌 클로라이드 및 에틸 아세테이트로 이루어진 군으로부터 선택된 1종이상의 용매에 구릿대 뿌리를 침지하여 상청액을 수득하거나 분획하는 것으로, 이후 증발 또는 동결건조를 통하여 분말화한 것이다. 상기 알콜로는 에탄올, 메탄올, 부탄올 또는 프로판올 등을 사용할 수 있으며, 바람직하기로는 메탄올이다. 그러나 상기 유기용매들은 상기 예시된 바에 한정되지 않으며, 공지의 모든 유기용매를 사용할 수 있다. Copper strip extract of the present invention is provided by extracting the copper bill with water or an organic solvent. Preferably, the supernatant is obtained by dipping copper roots in at least one solvent selected from the group consisting of alcohols having 1 to 5 carbon atoms, hexane, methylene chloride and ethyl acetate, and then powdered by evaporation or lyophilization. will be. Ethanol, methanol, butanol or propanol may be used as the alcohol, and preferably methanol. However, the organic solvents are not limited to the above examples, and all known organic solvents may be used.

가장 바람직한 구릿대 추출물 제조방법은 구릿대 뿌리 분쇄물을 메탄올에 1: 1 내지 30 중량비로 가하여 침지한 다음 이를 여과한 다음 감압, 농축 및 동결건조하여 구릿대 메탄올 추출물을 제조하고, 이후 헥산, 메틸렌 클로라이드 및 에틸 아세테이트를 순차적으로 가하여 각 용매별로 분획하여 헥산 분획, 메틸렌 클로라이드 분획 및 에틸 아세테이트 분획을 수득하는 것이다. 상기의 방법으로 수득한 구릿대 메탄올 추출물, 헥산 분획, 메틸렌 클로라이드 분획 및 에틸 아세테이트 분획은 모두 통칭하여 구릿대 추출물이라 이하 기재한다.The most preferred method for preparing copper leaf extract is immersed by adding copper leaf root crushed powder to methanol in a ratio of 1 to 1 to 30, and then filtering it, and then depressurizing, concentrating and lyophilizing to prepare a copper leaf methanol extract, and then hexane, methylene chloride and ethyl. Acetate is added sequentially to fractionate each solvent to obtain a hexane fraction, a methylene chloride fraction and an ethyl acetate fraction. The methanol extract, hexane fraction, methylene chloride fraction and ethyl acetate fraction obtained by the above method are all collectively referred to as copper extract.

본 발명의 구릿대 추출물은 프로스타글란딘 E2 합성을 억제하며, 특히 구릿대 메틸렌 클로라이드 분획이 가장 우수한 활성을 가진다.Copper leaf extract of the present invention inhibits prostaglandin E 2 synthesis, in particular copper leaf methylene chloride fraction has the best activity.

이후 본 발명에서는 구릿대 추출물로부터 프로스타글란딘 E2 합성 억제활성을 갖는 화합물을 분리하였다.Then, in the present invention, a compound having a prostaglandin E 2 synthesis inhibitory activity was isolated from the copper leaf extract.

본 발명의 프로스타글란딘 E2 합성 억제활성을 갖는 화합물은 화학식 1로 표시되는 화합물로, 그 예로는 하기 화학식 2의 펠로프테린(phellopterin), 화학식 3의 임페라토린(imperatorin), 화학식 4의 이소임페라토린(isoimperatorin), 화학식 5의 백안겔리신(byakangelicin), 화학식 6의 옥시퓨세다닌 메타놀레이트(oxypeucedanin methanolate), 화학식 7의 2',3'-디하이드로-8-하이드록시베르갑텐(2',3'-dihydro-8-hydroxybergapten), 화학식 8의 2',3'-디하이드로-8-하이드록시소랄렌(2',3'-dihydro-8-hydroxypsoralen), 화학식 9의 2',3'-디하이드로백안겔리신((2',3'-dihydrobyakangelicin), 화학식 10의 2',3'-디하이드옥시퓨세다닌 메타놀레이트((2',3'-dihydrooxypeucedanin methanolate) 및 화학식 11의 2',3',3,4-테트라하이드로백안겔리신이 있다.Compounds having a prostaglandin E 2 synthesis inhibitory activity of the present invention is a compound represented by the formula (1), for example, the felopterin (phellopterin) of the formula (2), imperatorin of the formula (3), isoimferatorin of the formula (4) (isoimperatorin), byakangelicin of formula 5, oxypeucedanin methanolate of formula 6, 2 ', 3'-dihydro-8-hydroxybergaptene of formula 7 (2', 3 '-dihydro-8-hydroxybergapten), 2', 3'-dihydro-8-hydroxysoralen of formula 8 (2 ', 3'-dihydro-8-hydroxypsoralen), 2', 3'- of formula 9 Dihydrobagangelicin ((2 ', 3'-dihydrobyakangelicin), 2', 3'-dihydroxyfucedanin methanolate of formula 10 ((2 ', 3'-dihydrooxypeucedanin methanolate) and 2' of formula 11, 3 ', 3,4-tetrahydrobackangelicin.

(화학식 2)(Formula 2)

(화학식 3)(Formula 3)

(화학식 4)(Formula 4)

(화학식 5)(Formula 5)

(화학식 6)(Formula 6)

(화학식 7)(Formula 7)

(화학식 8)(Formula 8)

(화학식 9)(Formula 9)

(화학식 10)Formula 10

(화학식 11)Formula 11

본 발명의 화학식 1로 표시되는 화합물은 프로스타글란딘 E2 합성 억제할 뿐만 아니라 COX-2 선택적 저해활성을 갖는 것으로 확인되어, 항염증제이나, 그 외 프로스타글란딘 E2 관련질환 또는 COX-2 관련질환에 대한 예방 및 치료제로 유용하게 사용할 수 있다.The compound represented by the formula (1) of the present invention was confirmed that not only inhibits prostaglandin E 2 synthesis but also has COX-2 selective inhibitory activity, thereby preventing and preventing proinflammatory agents, other prostaglandin E 2 related diseases, or COX-2 related diseases. It can be usefully used as a therapeutic agent.

이에, 본 발명은 구릿대 추출물 또는 화학식 1로 표시되는 화합물을 유효성분으로 포함하는 프로스타글란딘 E2 합성 억제제를 제공한다. 상기 프로스타글란딘 E2 합성 억제제는 프로스타글란딘 E2 관련 질환을 치료하기 위한 용도로 사용할 수 있으며, 가장 대표적인 용도는 소염제 또는 진통제이다. 또한, 상기 프로스타글란딘 E2 억제제는 혈관신생을 억제함으로 인한 암세포의 전이를 억제하는 작용과 더불어, 직장암, 대장암, 위암, 폐암 등 다양한 암세포의 세포사멸(아폽토시스)을 유도함으로서 항암제로 사용할 수 있다(Taketo MM (1998) J Natl Cancer Inst 90, 1529-1536, Eberhart CE et al (1994) Gastroenterrology, 107, 1183-1188, Sano H et al (1995) Cancer Res, 55, 3785-3789, Hwang D et al (1998) J Natl Cancer Inst. 90, 455-460). 따라서 본 발명의 프로스타글란딘 E2 합성 억제제는 진통제, 소염제 또는 항암제로의 가능성을 포함한다.Accordingly, the present invention provides a prostaglandin E 2 synthesis inhibitor comprising a copper extract or a compound represented by the formula (1) as an active ingredient. The prostaglandin E 2 synthesis inhibitor can be used for the treatment of prostaglandin E 2 related diseases, the most representative use is an anti-inflammatory or analgesic. In addition, the prostaglandin E 2 inhibitor may be used as an anticancer agent by inducing cell death (apoptosis) of various cancer cells such as rectal cancer, colorectal cancer, gastric cancer, lung cancer, in addition to inhibiting metastasis of cancer cells by inhibiting angiogenesis ( Taketo MM (1998) J Natl Cancer Inst 90, 1529-1536, Eberhart CE et al (1994) Gastroenterrology, 107, 1183-1188, Sano H et al (1995) Cancer Res, 55, 3785-3789, Hwang D et al (1998) J Natl Cancer Inst. 90, 455-460. Thus, the prostaglandin E 2 synthesis inhibitors of the present invention include the potential for analgesics, anti-inflammatory or anticancer agents.

본 발명의 프로스타글란딘 E2 합성 억제제는 경구 또는 비경구로 사용가능하며, 통상적인 의약품 제제인 경고제(PLASTERS), 과립제(GRANULES), 로션제(LOTIONS), 산제(POWDERS), 시럽제(SYRUPS), 액제(LIQUIDS AND SOLUTIONS), 에어로솔제(AEROSOLS), 연고제(OINTMENTS), 유동엑스제(FLUIDEXTRACTS), 유제(EMULSIONS), 현탁제(SUSPESIONS), 침제(INFUSIONS), 정제(TABLETS), 주사제(INJECTIONS), 캅셀제(CAPSULES) 또는 환제(PILLS) 형태를 제조할 수 있다.The prostaglandin E 2 synthesis inhibitors of the present invention can be used orally or parenterally, and are conventional pharmaceutical preparations such as PLASTERS, GRANULES, LOTIONS, POWDERS, Syrups, and SYRUPS. LIQUIDS AND SOLUTIONS, AEROSOLS, OINTMENTS, FLUIDEXTRACTS, Emulsions, SUPESIONS, INFUSIONS, TABLETS, INJECTIONS, CAPSULES or PILLS forms can be prepared.

본 발명의 프로스타글란딘 E2 합성 억제제는 약리학적으로 허용가능한 담체를 더욱 포함할 수 있다. 담체는 부형제, 결합제, 붕해제, 활택제, 향미제, 방부제, 안정제, 현탁제, 분산제, 희석제 또는 왁스로 사용되는 물질로, 일예로 셀룰로스, 인산칼슘, 탄산칼슘, 녹말, 카복시메틸 셀룰로스, 중탄산나트륨, 마그네슘 스테아레이트, 황산라우릴나트륨, 벤조에이트나트륨, 메틸파라벤, 시트르산, 암모늄 스테라에이트. 물, 덱스트린, 칼슘카보네이드, 락토스, 프로필렌글리톨, 리퀴드 파라틴, 젤라틴, 폴리에틸렌 글리콜, 올리브 오일, 에틸올레이트 및 생리식염수가 있다.Prostaglandin E 2 synthesis inhibitors of the invention may further comprise a pharmacologically acceptable carrier. Carriers are materials used as excipients, binders, disintegrants, lubricants, flavors, preservatives, stabilizers, suspending agents, dispersants, diluents or waxes, for example cellulose, calcium phosphate, calcium carbonate, starch, carboxymethyl cellulose, bicarbonate Sodium, magnesium stearate, sodium lauryl sulfate, sodium benzoate, methylparaben, citric acid, ammonium stearate. Water, dextrin, calcium carbonate, lactose, propylene glycol, liquid paratin, gelatin, polyethylene glycol, olive oil, ethyl oleate and physiological saline.

프로스타글란딘 E2 합성 억제제의 투여량은 환자의 연령, 성별, 상태, 체내에서 활성 성분의 흡수도, 불활성율 및 병용되는 약물을 고려하여 적절히 조절할 수 있으며, 일예로 유효성분 0.01 내지 500 ㎎/㎏의 단일 투여량으로 하루 1 내지 3회 투여될 수 있다.The dosage of the prostaglandin E 2 synthesis inhibitor may be appropriately adjusted in consideration of the age, sex, condition, absorption of the active ingredient, inactivation rate and the drug used in the body of the patient, for example, an active ingredient of 0.01 to 500 mg / kg. A single dose may be administered 1-3 times a day.

또한 상기 기재한 프로스타글란딘 E2 합성 억제제는 프로스타글란딘 E2 관련질환 치료 및 예방용 약학적 조성물로 제공가능하다. 상기 관련질환은 염증관련 질환 및 암이 있다.In addition, the prostaglandin E 2 synthesis inhibitors described above can be provided as a pharmaceutical composition for treating and preventing prostaglandin E 2 related diseases. The related diseases include inflammatory diseases and cancer.

이하 본 발명의 실시예를 기재한다. 하기 실시예는 본 발명을 예시하기 위한 것일 뿐 본 발명이 하기 실시예에 한정되는 것은 아니다. Hereinafter, examples of the present invention will be described. The following examples are only for illustrating the present invention and the present invention is not limited to the following examples.

실시예 1: 구릿대 추출물 제조 및 PGEExample 1 preparation of copper leaf extract and PGE 22 생성 저해효과 검정 Production inhibitory effect test

1-1. 구릿대 추출물 제조1-1. Copper leaf extract manufacturer

구릿대 뿌리인 백지를 경도시장에서 200년 9월에 구입한 후 서울대학교 천연물 연구소로부터 검증 받은 다음 실험에 사용하였다.White paper, a copper root, was purchased at the hardness market in September, 200, and was used in the following experiment after it was verified by the Natural Products Research Institute of Seoul National University.

백지 분말 3 ㎏을 메탄올 8 ℓ에 침지한 후 총 4시간 환류시켜 3회추출하였다. 추출액은 모두 수집하고 감압농축하여 520 g의 메탄올 추출물을 수득하였다.3 kg of white paper powder was immersed in 8 L of methanol and refluxed three times for 4 hours. The extracts were collected and concentrated under reduced pressure to give 520 g of methanol extract.

메탄올 추출물에 n-헥산:메탄올:물을 10:1:9로 가한 후 분획하여 헥산 분획을 수득하였다.N-hexane: methanol: water was added to the methanol extract at 10: 1: 9 and fractionated to obtain a hexane fraction.

이후 잔류물에 메틸렌 클로라이드 및 에틸 아세테이트를 순차적으로 가하여 메틸렌 클로라이드 분획, 에틸 아세테이트 및 물 분획을 각각 수득하였다.Methylene chloride and ethyl acetate were then sequentially added to the residue to obtain methylene chloride fraction, ethyl acetate and water fraction, respectively.

1-2. 구릿대 추출물의 PGE2 생성 저해능 검증1-2. Validation of PGE 2 Production Inhibition of Copper Leaf Extract

공지된 방법(Ohuchi K, Watanabe M, Yoshizawa K, Tsurufuji S, Fujiki H, Suganuma M, Sugimura T, Levine L. (1985) Biochimica et Biophysica Acta, 834: 42-7)으로 복막세포를 스프레그-다울리 쥐(Sprague-Dawley rat)로부터 수득하였다.Peritoneal cells are spread by known methods (Ohuchi K, Watanabe M, Yoshizawa K, Tsurufuji S, Fujiki H, Suganuma M, Sugimura T, Levine L. (1985) Biochimica et Biophysica Acta, 834: 42-7). Obtained from Sprague-Dawley rats.

복막세포는 10 % 소 혈청(Flow Laboratories, North Rydge, Australia), 페니실린 G 포타슘 18 ㎍/㎖ 및 스트렙토마이신 설페이트 50 ㎍/㎖(Meiji Seika, Tokyo, Japan)을 포함하는 EMEM(Eagle's minimal essential medium, Nissui, Tokyo, Japan)에 현탁한 후 24-웰 플라스틱 세포 배양판(Corning Glass Works, Corning, NY, USA)에 7.5 x 105 cells/0.5 ㎖/웰로 넣고 37 ℃에서 2시간 배양하였다. 이후 배지로 3회 세척하여 부착되지 않은 세포들은 제거한 후 37 ℃에서 20시간 더욱 배양하였다. 3회 세척한 후 부착된 세포들은 다음 실험에 이용하였다.Peritoneal cells contained 10% bovine serum (Flow Laboratories, North Rydge, Australia), 18 μg / ml penicillin G potassium and 50 μg / ml streptomycin sulfate (Meiji Seika, Tokyo, Japan). After suspension in Nissui, Tokyo, Japan), the cells were placed in a 24-well plastic cell culture plate (Corning Glass Works, Corning, NY, USA) at 7.5 × 10 5 cells / 0.5 mL / well and incubated at 37 ° C. for 2 hours. After washing three times with medium to remove the non-attached cells were further incubated for 20 hours at 37 ℃. After washing three times, the attached cells were used in the next experiment.

부착된 세포들은 10 % 소 혈청을 포함하는 배지 0.5 ㎖상에서 37 ℃로 8시간 배양하였다. 또한 각 세포에 LPS 0.1 ㎍/㎖과 구릿대 추출물(메탄올 추출물, 헥산 분획, 메틸렌 클로라이드 분획, 에틸 아세테이트 분획 및 물 분획) 30 ㎍/㎖을 각각 또는 함께 처리하여 배양하였고, 양성대조군으로 인도메타신(Sigma Chemical Co., St. Louis, MO, USA)을 에탄올에 용해시켜 0.1 μM로 사용하였다. Attached cells were incubated at 37 ° C. for 8 hours on 0.5 ml of medium containing 10% bovine serum. In addition, each cell was incubated with 0.1 μg / ml of LPS and 30 μg / ml of copper leaf extract (methanol extract, hexane fraction, methylene chloride fraction, ethyl acetate fraction and water fraction), respectively or together, and treated with indomethacin as a positive control group. Sigma Chemical Co., St. Louis, MO, USA) was dissolved in ethanol and used at 0.1 μM.

배양후, 배지만을 수거한 후 5000 xg로 4 ℃에서 5분간 원심분리하였고, 상등액내 PGE2의 농도를 공지의 방사면역분석법(Ohuchi K, Watanabe M, Yoshizawa K, Tsurufuji S, Fujiki H, Suganuma M, Sugimura T, Levine L. (1985) Biochimica et Biophysica Acta, 834: 42-7)으로 확인하였다. 이때 사용한 PGE2 항-혈청은 Assay Designs(Ann Arbor, MI, USA)사로부터 구입하였다.After incubation, only medium was collected and centrifuged at 5000 xg for 5 minutes at 4 ° C. The concentration of PGE 2 in the supernatant was determined by known radioimmunoassays (Ohuchi K, Watanabe M, Yoshizawa K, Tsurufuji S, Fujiki H, Suganuma M). , Sugimura T, Levine L. (1985) Biochimica et Biophysica Acta, 834: 42-7). The PGE 2 antiserum used at this time was purchased from Assay Designs (Ann Arbor, MI, USA).

통계처리는 다중비교는 두넷 테스트(Dunnett's test)를 사용하였고, 개별 관찰시에서 Student's t-test로 하였다.For statistical analysis, Dunnett's test was used for multiple comparison, and Student's t- test was used for each observation.

실험군Experimental group PGE2(ng/㎖)PGE 2 (ng / ml) 무처리군No treatment group 2.07 ± 0.15*** 2.07 ± 0.15 *** LPS 0.1 ㎍/㎖0.1 μg / ml LPS 8.40 ± 0.158.40 ± 0.15 LPS + 메탄올 추출물(30 ㎍/㎖)LPS + Methanol Extract (30 μg / mL) 5.61 ± 0.21*** 5.61 ± 0.21 *** LPS + 헥산 분획(30 ㎍/㎖)LPS + Hexane Fraction (30 μg / mL) 2.53 ± 0.11*** 2.53 ± 0.11 *** LPS + 메틸렌 클로라이드 분획(30 ㎍/㎖)LPS + methylene chloride fraction (30 μg / ml) 1.96 ± 0.13*** 1.96 ± 0.13 *** LPS + 에틸 아세테이트 분획(30 ㎍/㎖)LPS + ethyl acetate fraction (30 μg / ml) 6.54 ± 0.28** 6.54 ± 0.28 ** LPS + 물 분획(30 ㎍/㎖)LPS + water fraction (30 μg / ml) 8.40 ± 0.068.40 ± 0.06 LPS + 인도메타신(0.1 μM)LPS + Indomethacin (0.1 μM) 1.89 ± 0.17*** 1.89 ± 0.17 *** **:P<0.01,***: P<0.001 vs. LPS 처리군 ** : P <0.01, *** : P <0.001 vs. LPS treatment group

각 구릿대 추출물이 대식세포에서의 PGE2 생산에 미치는 영향을 확인한 결과는 상기 표 1과 같다. 즉, 복막세포에 LPS를 처리하여 유도된 PGE2는 물 분획을 제외한 각 구릿대 추출물에 처리에 의하여 생산이 현저히 저해되었다. 특히 구릿대 메틸렌 클로라이드 분획의 저해효과가 가장 우수하였으며, 거의 양성대조군인 인도메탄신을 처리한 경우와 유사한 결과를 나타내었다.The results of confirming the effect of each copper extract on PGE 2 production in macrophages are shown in Table 1 above. That is, PGE 2 induced by LPS treatment of peritoneal cells was significantly inhibited by the treatment of extracts of copper strips except water fraction. In particular, the inhibitory effect of the copper-methylene chloride fraction was the best and showed similar results to the treatment of indomethane, an almost positive control group.

실시예 2: 화합물 분리/제조 및 PGEExample 2: Compound Separation / Preparation and PGE 22 생성 저해효과 검정 Production inhibitory effect test

2-1. 구릿대 추출물로부터 활성 화합물 분리2-1. Isolation of Active Compounds from Copper Extract

실시예 1와 동일한 방법으로 헥산 분획을 제조하였고, 헥산 분획 수득후 잔류물에 에틸아세테이트를 가하여 에틸아세테이트 분획을 수득하였으며, 상기 두 분획으로부터 각각 활성 화합물을 분리 및 동정하였다.The hexane fraction was prepared in the same manner as in Example 1, and ethyl acetate was added to the residue to obtain the hexane fraction, thereby obtaining an ethyl acetate fraction. The active compounds were separated and identified from the two fractions, respectively.

헥산 분획 30.4 g을 실리카겔이 충진된 컬럼(70-230 mesh, Art, 7734 Merck)에 흡착시킨 후 헥산 및 디에틸에테르 0 내지 100 % 농도구배를 갖는 용매를 컬럼에 흘려주어 용출된 화합물 각각을 분리하였고, 분광 데이터(NMR, IR, UV)를 이용하여 동정하여 펠로프테린(phellopterin, 화학식 2) 0.98 g, 임페라토린(imperatorin, 화학식 3) 2.88 g 및 이소임페라토린(isoimperatorin, 화학식 4) 1.68 g을 각각 수득하였다.30.4 g of the hexane fraction was adsorbed onto a silica gel-filled column (70-230 mesh, Art, 7734 Merck), and then a solvent having a concentration gradient of hexane and diethyl ether from 0 to 100% was poured through the column to separate the eluted compounds. It was identified using spectral data (NMR, IR, UV), 0.98 g of pepopterin (Formula 2), 2.88 g of Imperatorin (Formula 3) and 1.68 g of Isoimperatorin (Formula 4). Were obtained respectively.

또한 에틸아세테이트 분획 75.3 g을 실리카겔이 충진된 컬럼(70-230 mesh, Art, 7734 Merck)에 흡착시킨 후 메틸렌 클로라이드 및 메탄올 0 내지 20 % 농도구배를 갖는 용매를 컬럼에 흘려주어 용출된 화합물 각각을 분리하였고, 분광 데이터(NMR, IR, UV)를 이용하여 동정하여 백안겔리신(byakangelicin, 「α」D 20: + 17.5°c 0.10, C5H5N), 화학식 5) 3.69 g과 옥시퓨세다닌 메타놀레이트(oxypeucedanin methanolate, 「α」D 20: + 18.0°c 0.20, CHCl 3), 화학식 6) 0.11 g을 수득하였다.In addition, 75.3 g of ethyl acetate fraction was adsorbed onto a silica gel-filled column (70-230 mesh, Art, 7734 Merck), and methylene chloride and a solvent having a concentration gradient of 0 to 20% methanol were poured through the column, thereby eluting each of the eluted compounds. Was isolated and identified using spectral data (NMR, IR, UV) by white angelicin (byakangelicin, "α" D 20 : + 17.5 ° c 0.10, C 5 H 5 N), 3.69 g of Formula 5) and oxyfuse 0.11 g of danin methanol (oxypeucedanin methanolate, "α" D 20 : + 18.0 ° C 0.20, CHCl 3 ), formula 6) was obtained.

2-2. 분리한 활성 화합물의 반합성물 합성2-2. Semisynthetic Synthesis of Isolated Active Compounds

상기에서 분리한 5종의 화합물들로부터 반합성 화합물 5종을 각각 합성하였다.Five semi-synthetic compounds were synthesized from the five compounds separated above.

상기에서 분리한 5종의 화합물들은 퓨라노쿠마린계 화합물로, 퓨라노쿠마린계 화합물은 퓨란 고리와 피라노-2-온 고리에 각각 하나의 이중결합을 가지고 있다. 상기 이중결합은 10 % 팔라듐/차콜(Pd/C)의 몰비율을 조절하여 선택적으로 수소화시킬 수 있다. The five compounds separated above are furanocoumarin compounds, and the furanocoumarin compounds each have one double bond in the furan ring and the pyrano-2-one ring. The double bond may be selectively hydrogenated by controlling the molar ratio of 10% palladium / charcoal (Pd / C).

퓨란 고리내 이중결합을 선택적으로 수소화시키기 위하여, Pd/C 및 퓨라노쿠마린 화합물(펠로프테린, 임페라토린, 이소임페라토린, 백안겔리신 또는 옥시퓨세다닌 메타놀레이트)을 1:1 몰 비율로 에탄올(1 g/50 ㎖)에 용해시키고, 상온에서 수소가스하에서 30분간 충분히 혼합하였다. 혼합물에서 반합성된 화합물 화학식 7의 2',3'-디하이드로-8-하이드록시베르갑텐(2',3'-dihydro-8-hydroxybergapten), 화학식 8의 2',3'-디하이드로-8-하이드록시소랄렌(2',3'-dihydro-8-hydroxypsoralen), 화학식 9의 2',3'-디하이드로백안겔리신((2',3'-dihydrobyakangelicin, 「α」D 20: + 18.5°c 0.30, C5H5 N)) 및 화학식 10의 2',3'-디하이드옥시퓨세다닌 메타놀레이트((2',3'-dihydrooxypeucedanin methanolate, 「α」D 20: + 19.0°c 0.25, CHCl3))를 각각 분리하였다.In order to selectively hydrogenate the double bond in the furan ring, ethanol of Pd / C and furanocoumarin compound (felloperin, imperatorin, isoimperatorine, white angelicin or oxyfusedin methanolate) in a 1: 1 molar ratio It was dissolved in (1 g / 50 mL) and mixed well for 30 minutes under hydrogen gas at room temperature. Compounds Semi-Synthesized in Mixture 2 ', 3'-dihydro-8-hydroxybergapten of Formula 7, 2', 3'-dihydro-8 of Formula 8 -hydroxy small ralren (2 ', 3'-dihydro- 8-hydroxypsoralen), 2 of formula IX', 3'-dihydro the bag gelri new ((2 ', 3'-dihydrobyakangelicin , "α" D 20: + 18.5 ° c 0.30, C 5 H 5 N)) and 2 ', 3'-dihydroxyfucedanin methanolate of formula 10 ((2', 3'-dihydrooxypeucedanin methanolate, "α" D 20 : + 19.0 ° c 0.25, CHCl 3 )), respectively.

또한, 2개의 이중결합을 모두 수소화하기 위하여 Pd/C 및 백안겔리신을 2:1 몰비율로 에탄올에 용해시킨 후 상기와 동일한 방법으로 실시하여 화학식 11의 2',3',3,4-테트라하이드로백안겔리신(「α」D 20: + 17.5°c 0.30, C5 H5N))을 수득하였다.In addition, in order to hydrogenate both double bonds, Pd / C and white angelicin were dissolved in ethanol at a 2: 1 molar ratio, and the same procedure as in the above was carried out to give 2 ′, 3 ′, 3,4-tetra of Formula 11 dihydro the bag gelri Shin: a ( "α" D 20 + 17.5 ° c 0.30, C 5 H 5 N)) was obtained.

2-3. PGE2 생산 저해율 검정2-3. PGE 2 Production Inhibition Rate Assay

상기에서 분리 및 반합성한 총 10종의 화합물들의 PGE2 생산 저해율을 측정하였다. 실험방법은 실시예 1-2와 동일하게 실시하였고, 다만 10종의 화합물들 각각은 DMSO(dimethyl sulfoxide)에 용해시켜 30 μM로 배지에 첨가하였다. 양성대조군으로 인도메타신(Sigma Chemical Co., St. Louis, MO, USA)을 에탄올에 용해시켜 사용하였고, 배지내 DMSO 및 에탄올의 총 농도는 0.1 (v/v)%이다.The inhibition of PGE 2 production of a total of 10 compounds isolated and semisynthesized was measured. Experimental method was carried out in the same manner as in Example 1-2, except that each of the 10 compounds were dissolved in DMSO (dimethyl sulfoxide) and added to the medium at 30 μM. As a positive control, indomethacin (Sigma Chemical Co., St. Louis, MO, USA) was used in ethanol, and the total concentration of DMSO and ethanol in the medium was 0.1 (v / v)%.

그 결과는 도 1에 도시하였다. 도 1은 쥐 복막의 대식세포에서 LPS로 유도된 PGE2 생산에 있어 10 종의 퓨라노쿠마린계 화합물이 미치는 영향을 나타낸 것으로(**: P<0.01, ***: P<0.001 vs. LPS 처리군), 1 내지 10은 각각 백안겔리신, 2',3'-디하이드로백안겔리신, 2',3',3,4-테트라하이드로백안겔리신, 펠로프테린, 2',3'-디하이드로-8-하이드록시베르갑텐, 임페라토린, 2',3'-디하이드로-8-하이드록시소랄렌, 이소임페라토린, 옥시퓨세다닌 메타놀레이트 및 2',3'-디하이드옥시퓨세다닌 메타놀레이트를 처리한 것이다.The result is shown in FIG. FIG. 1 shows the effect of 10 furanocoumarin compounds on LPS-induced PGE 2 production in macrophages of rat peritoneum (**: P <0.01, ***: P <0.001 vs. LPS) Treatment group), 1 to 10 are white angelicin, 2 ', 3'-dihydroback angelicin, 2', 3 ', 3,4-tetrahydrobackangelicin, pelopterin, 2', 3 ', respectively. -Dihydro-8-hydroxybergaptene, imperatorin, 2 ', 3'-dihydro-8-hydroxysoralen, isoimperatorine, oxyfusedin methanolate and 2', 3'-dihydroxyfuse Treated with methanol metholate.

도 1에서, 10종의 퓨라노쿠마린계 화합물은 PGE2 생성을 억제하였으며, 특히 백안겔리신, 2',3'-디하이드로백안겔리신, 펠로프테린, 임페라토린, 2',3'-디하이드로-8-하이드록시소랄렌, 이소임페라토린 및 옥시퓨세다닌 메타놀레이트의 저해율이 매우 우수하였고, 펠로프테린 및 임페라토린이 가장 높은 저해율을 나타내어 양성대조군인 인도메타신과 거의 유사하거나 좀더 우수하였다.In Fig. 1, 10 kinds of furanocoumarin compounds inhibited PGE 2 production, in particular white angelicin, 2 ', 3'-dihydrobackangelicin, pelophterin, imperatorin, 2', 3'- The inhibition rate of dihydro-8-hydroxysorylene, isoimferatorin and oxyfusedin methanolate was very good, and the pelopterin and imperatorin showed the highest inhibition rate, which was almost similar to or better than the positive control indomethacin. .

또한 구릿대 뿌리로부터 분리한 5종의 퓨라노쿠마린계 화합물이 반합성한 5종의 화합물에 비하여 PGE2 저해활성이 우수한 것으로 관찰되어, 이는 퓨란 고리 및 피라노-2-온 고리상에 존재하는 이중결합이 PGE2 저해에 중요한 역할을 하는 것으로 생각된다.In addition, it was observed that PUR 2 inhibitory activity was superior to the five compounds synthesized by the five kinds of furanocoumarin-based compounds isolated from the roots of the copper leaf, which are double bonds on the furan ring and the pyrano-2-one ring. It is thought to play an important role in this inhibition of PGE 2 .

실시예 3: 활성 화합물의 PGEExample 3: PGE of Active Compound 22 생성 저해기작 검정 Production Inhibitory Mechanism Assay

활성 화합물에 의한 PGE2 생성 저해기작을 확인하기 위하여, 임페라토린이 COX-2 및 mPGES의 발현율에 미치는 영향을 조사하였다.In order to confirm the mechanism of inhibition of PGE2 production by the active compounds, the effects of imperatorin on the expression rate of COX-2 and mPGES were investigated.

먼저, [3H]아라키도닌 산-표지된 대식세포(Ohuchi K, Sugawara T, Watanabe M, Hirasawa N, Tsurufuji S, Fujiki H, Christensen SB, Sugimura T. (1998) British Journal of Pharmacology, 94: 917-23)에 LPS 및 임페라토린을 처리한 후 배지를 수거하여 방사능을 측정하였다. 그 결과 LPS 단독 처리시 측정된 방사능 값이 매우 증가되어, 인지질막으로부터 아라키도닌산이 방출됨을 확인할 수 있었다. 그러나, LPS와 함께 임페라토린을 처리한 경우에도 측정된 방사능 값이 LPS 단독 처리시와 유사한 것으로 측정되어, 임페라토린이 아라키도닌산의 인지질막으로부터의 방출을 저해하는 것은 아님을 알 수 있었다.First, [ 3 H] arachidonin-labeled macrophages (Ohuchi K, Sugawara T, Watanabe M, Hirasawa N, Tsurufuji S, Fujiki H, Christensen SB, Sugimura T. (1998) British Journal of Pharmacology, 94: 917-23) was treated with LPS and imperatorin, and then the medium was collected for radioactivity. As a result, it was confirmed that the radioactivity value measured when the LPS alone treatment was increased so that arachidonic acid was released from the phospholipid membrane. However, even when treated with impertorin with LPS, the measured radioactivity value was similar to that of LPS alone treatment, indicating that imperatorin did not inhibit the release of arachidonic acid from the phospholipid membrane.

따라서, 방출된 아라키도닌산이 PGE2로 전환되는 단계에서 임페라토린이 작용하는지 여부를 하기 실험으로 확인하였다.Therefore, it was confirmed by the following experiment whether or not the imperatorin action in the step of converting the released arachidonic acid to PGE 2 .

복막 세포 3 x 106를 실시예 1과 동일한 방법으로 준비한 후 각 시료가 처리된 배지 2 ㎖을 첨가한 후 37 ℃에서 8시간 배양하였다. 사용한 시료는 LPS 0.1 ㎍/㎖, 덱사메타손(dexamethasone: DEX) 0. μM, 임페라토린 (3, 10 또는 30 μM)이며, 덱사메타손과 임페라토린은 LPS와 함께 사용하였다. 배양한 세포로부터 세포질 분획을 수득한 후 이를 SDS-폴리아크릴아마이드 젤에 전기영동하였고, COX-1 항체(Santa Cruz Biotechnology, CA, USA), COX-2 항체(Santa Cruz Biotechnology), cPGES 항체(Cayman Chemical Co., Ann Arbor, MI, USA) 및 mPGES 항체(Cayman Chemical Co.) 각각을 프로브로 사용하여 웨스턴 블롯을 실시하였다(Kim YP, Yamada M, Lim SS, Lee SH, Ryu N, Shin KH, Ohuchi K. (1999) Biochimica et Biophysica Acta, 1438:399-407).Peritoneal cells 3 × 10 6 were prepared in the same manner as in Example 1, followed by addition of 2 ml of medium in which each sample was treated, followed by incubation at 37 ° C. for 8 hours. Samples used were 0.1 μg / ml LPS, dexamethasone (DEX) 0.1 μM, imperatorin (3, 10 or 30 μM), and dexamethasone and imperatorin were used with LPS. Cytoplasmic fractions were obtained from the cultured cells and electrophoresed on SDS-polyacrylamide gels, and COX-1 antibodies (Santa Cruz Biotechnology, CA, USA), COX-2 antibodies (Santa Cruz Biotechnology), and cPGES antibodies (Cayman). Western Co., Ltd. (Kim YP, Yamada M, Lim SS, Lee SH, Ryu N, Shin KH, using Chemical Co., Ann Arbor, MI, USA) and mPGES antibody (Cayman Chemical Co.), respectively, as probes. Ohuchi K. (1999) Biochimica et Biophysica Acta, 1438: 399-407.

도 2는 LPS 처리된 복막 대식세포에서의 COX-1, COX-2, cPGES 및 mPGES의 발현에 있어 임페라토린의 영향을 확인한 것으로, A는 웨스턴 블롯 사진이고, B는 임페라토린의 PGE2 생성 저해율을 나타낸 것이다(*** P <0.001 vs. LPS 처리군).Figure 2 confirms the effect of imperatorin on the expression of COX-1, COX-2, cPGES and mPGES in LPS-treated peritoneal macrophages, A is a Western blot picture, B is the inhibition rate of impertorin PGE 2 production (*** P <0.001 vs. LPS treated group).

도 2A를 살펴보면, 복막 대식세포는 COX-1, COX-2, cPGES 및 mPGES를 일정수준으로 발현하고 있으나 LPS 처리시 COX-2 및 mPGES을 보다 현저히 발현시킨다. 특히 COX-2의 발현량이 급격히 증가한다. 여기에 덱사메타손을 처리하면 COX-2와 mPGES의 발현정도가 현저히 억제되며, 또한 임페라토린 역시 농도에 의존적인 양상으로 COX-2 및 mPGES 발현을 억제시킨다. 임페라토린은 도 2B에 나타낸 바와 같이 농도에 비례하여 PGE2 발현을 저해한다. 상기한 결과를 토대로, 임페라토린의 PGE2 생성 저해는 COX-2 및 mPGES의 발현을 억제하므로써 이루어짐을 알 수 있다. 상기 COX-2 및 mPGES는 초기 염증 반응 촉진자들에 의하여 발현되는 것으로, 염증반응과 관련있는 효소들이다. 따라서, 본 발명의 퓨라노쿠마린계 화합물은 항염증제로 사용가능하다.Referring to Figure 2A, peritoneal macrophages express COX-1, COX-2, cPGES and mPGES at a certain level, but more significantly express COX-2 and mPGES during LPS treatment. In particular, the expression level of COX-2 increases rapidly. Dexamethasone treatment significantly inhibits the expression level of COX-2 and mPGES, and imperatorin also inhibits COX-2 and mPGES expression in a concentration-dependent manner. Imperatorin inhibits PGE 2 expression in proportion to concentration as shown in FIG. 2B. Based on the above results, it can be seen that inhibition of PGE 2 production by imperatorin is achieved by inhibiting the expression of COX-2 and mPGES. The COX-2 and mPGES are expressed by early inflammatory response promoters and are enzymes involved in the inflammatory response. Therefore, the furanocoumarin compound of the present invention can be used as an anti-inflammatory agent.

이상 살펴본 바와 같이, 본 발명의 구릿대 추출물 또는 이로부터 유래한 퓨라노쿠마린계 화합물들은 프로스타글란딘 E2 합성을 억제하므로써 소염제, 진통제 또한 항암제로 사용가능하며, 특히 임페라토린은 COX-2에 선택적 저해성을 가져 기존의 비스테로이드성계 소염제에 비하여 안전하게 사용할 수 있다.As described above, the copper leaf extract of the present invention or furanocoumarin-based compounds derived therefrom can be used as an anti-inflammatory agent, an analgesic agent as an anticancer agent by inhibiting prostaglandin E 2 synthesis, and especially imperatorin has a selective inhibitory effect on COX-2. It can be used safely compared to existing nonsteroidal anti-inflammatory agents.

도 1은 쥐 복막의 대식세포에서 LPS로 유도된 PGE2 생산에 있어 10 종의 화합물이 미치는 영향을 나타낸 것이다.Figure 1 shows the effect of 10 compounds on LPS-induced PGE 2 production in rat peritoneal macrophages.

도 2는 LPS 처리된 복막 대식세포에서의 COX-1, COX-2, cPGES 및 mPGES의 발현에 있어 임페라토린의 영향을 확인한 것이다.Figure 2 confirms the effect of imperatorin on the expression of COX-1, COX-2, cPGES and mPGES in LPS treated peritoneal macrophages.

Claims (6)

하기 화학식 1로 표시되는 화합물을 유효성분으로 포함하는 프로스타글란딘 E2 합성 억제제:Prostaglandin E 2 synthesis inhibitor comprising a compound represented by the formula (1) as an active ingredient: (화학식 1)(Formula 1) 상기 화학식 1에서,In Chemical Formula 1, R1은 H, OCH3, OCH2-CH=C(CH3)2 또는 이고;R 1 is H, OCH 3 , OCH 2 —CH═C (CH 3 ) 2 or ego; R2는 H, OCH2-CH=C(CH3)2 또는 이고; 및R 2 is H, OCH 2 —CH═C (CH 3 ) 2 or ego; And R3-R4 및 R5-R6은 각각 독립적으로 HC=CH 또는 H2 C-CH2 이다.R 3 -R 4 and R 5 -R 6 are each independently HC═CH or H 2 C—CH 2 . 제 1항에 있어서, 상기 화합물은 하기 화학식 2 내지 11로 표시되는 화합물로 이루어진 군으로부터 선택되는 것인 억제제:The inhibitor according to claim 1, wherein the compound is selected from the group consisting of compounds represented by the following Chemical Formulas 2 to 11. (화학식 2)(Formula 2) (화학식 3)(Formula 3) (화학식 4)(Formula 4) (화학식 5)(Formula 5) (화학식 6)(Formula 6) (화학식 7)(Formula 7) (화학식 8)(Formula 8) (화학식 9)(Formula 9) (화학식 10)Formula 10 (화학식 11)Formula 11 구릿대 추출물을 유효성분으로 포함하는 프로스타글란딘 E2 합성 억제제.Prostaglandin E 2 synthesis inhibitor comprising a copper leaf extract as an active ingredient. 제 4항에 있어서, 상기 구릿대 추출물은 구릿대 뿌리를 메탄올, 헥산, 메틸렌 클로라이드 및 에틸 아세테이트로 이루어진 군으로부터 1종이상 선택되는 용매로 추출하여 제조한 것인 억제제.The inhibitor according to claim 4, wherein the copper leaf extract is prepared by extracting copper leaf roots with a solvent selected from at least one solvent selected from the group consisting of methanol, hexane, methylene chloride and ethyl acetate. 제 1항 또는 제 3항의 프로스타글란딘 E2 합성 억제제를 포함하는 프로스타글란딘 E2 관련 질환 예방 및 치료용 약학적 조성물.Claim 1 or claim 3 prostaglandin E 2, prostaglandin E 2 synthesis inhibitors, including the prevention and treatment of diseases related to the pharmaceutical composition. 제 5항에 있어서, 상기 약학적 조성물은 소염제, 진통제 또는 항암제인 약학적 조성물.6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition is an anti-inflammatory, analgesic or anticancer agent.
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