발간등록번호 11-1543000-000646-01 오가피및문주란추출물복합제를이용한발모제개발
농림축산식품부장관귀하 이보고서를 오가피및문주란추출물복합제를이용한발모제개발 과제의보고서로 제출합니다. 2014 년 9 월 5 일 주관연구기관명 : 삼천당제약주관연구책임자 : 임광진협동연구기관명 : 제주대협동연구책임자 : 강희경
Ⅰ. 제목 오가피및문주란추출물복합제를이용한발모제개발 Ⅱ. 연구개발의목적및필요성현재사용되고있는미녹시딜과피나스테라이드등의발모제는충분한효과를보여주지못하고있을뿐만아니라, 발기부전, 저혈압환자의혈압저하, 임부복용시남성태아외부생식기의비정상초래등의심각한부작용을나타내장기간사용하기에는부적합하다고보고되고있음. 따라서발모효과가확인된오가피와문주란추출물의복합제를제조하여화학성분의기존발모제보다효과는우수하고부작용은개선된천연물발모제를개발하고자함 Ⅲ. 연구개발내용및범위 - 오가피와문주란에서발모에효과적인활성분획추출 - 활성분획추출체계확립 - 발모제제조공정확립 - 발모제효능실험 - 제제연구 - 기전연구 - 안전성시험 - 탈모치료에효과적인발모제개발 Ⅳ. 연구개발결과 - 오가피와문주란추출물복합제의표준제조공정확립및지표성분도출 - 기존제품인 5% minoxidil과동등한효능을발휘하는천연물발모제개발기술확보 - 기전연구결과오가피와문주란추출물복합제가 dermal papilla cells에서모발의성장에관여하는 wnt/β-catenin pathway 활성화및세포주기진행에관여하는단백질의발현을조절하여세포성장유도하는것을확인 - 단회투여독성시험, 4주 DRF 시험, 항원성시험, 피부자극시험, 안점막자극시험완료 Ⅴ. 연구성과및성과활용계획 - 오가피와문주란추출물복합제의장기독성시험을진행하여확보된안전성시험자료를토대로 IND 허가신청예정 - 본연구성과를바탕으로발모제조성물특허출원을완료하였으며, PCT 출원을진행중에있어획득한기술및정보를활용하여향후천연물신약개발에응용계획 - 발모촉진에효능을나타내는국내자생식물의활용으로재배농가의소득증대기여 - 삼천당제약의병원및약국유통망을이용하여제품유통및판매
SUMMARY Ⅰ. Title Development of hair growth solutions from Acanthopanax koreanum and Critinum asiaticum complex extracts Ⅱ. Purpose and necessity of research and development The hair growth drugs on the market, minoxidil and finasteride, have no satisfactory potency, but serious side effects such as erectile dysfunction, blood pressure drops in hypotensive patients, and abnormal external genitalia of male fetus in pregnant women. Therefore, long-term use of these drugs is not recommended. We started to develop a more effective and safer than the current hair growth drugs using extracts from Acanthopanax koreanum and Critinum asiaticum which had hair growth effects. Ⅲ. Content and scope of research and development - Establishment of the extraction process of hair growth fraction - Establishment of the manufacturing process of hair growth fraction - Efficacy study - Formulation study - Drug mechanism study - Safety study - Development of hair growth solutions Ⅳ. Result of research and development - Establishment of the manufacturing process and the marker compounds for Acanthopanax koreanum and Critinum asiaticum complex extracts - Development of Acanthopanax koreanum and Critinum asiaticum complex extracts having the hair regrowth promoting effects similar to 5% minoxidil in animal studies - Finding that Acanthopanax koreanum and Critinum asiaticum complex extracts induce hair cell proliferation through the regulation of protein expression involved in the cell cycle and activation of wnt/β-catenin pathway involved in hair growth in dermal papilla cells - Completion of single dose toxicity study, repeated dose 4 week dose-range finding toxicity study, antigenicity study, skin irritation test, and eye irritation test Ⅴ. Future plan for the utillization of research and development - IND submission after long-term repeated dose toxicity studies of Acanthopanax koreanum and Critinum asiaticum complex extracts - Patent Application in Korea and PCT application - Contribution to Korean farmers' income by using Korean native plants - Marketing the natural hair growth drug through the hospitals and pharmacies related with SCD Pharm.
CONTENTS Chapter 1. Outlines of Research Project Chapter 2. Domestic and International Research Status Chapter 3. Contents of the Research Project and Results Chapter 4. Level of Achievement and Contribution to Related Areas Chapter 5. Application of Research Results Chapter 6. Research Facility and Machinery Status
제 1 장연구개발과제의개요 제 2 장국내외기술개발현황 제 3 장연구개발수행내용및결과 제 4 장목표달성도및관련분야에의기여도 제 5 장연구개발성과및성과활용계획 제 6 장연구시설장비 현황
제 1 장 연구개발과제의개요 - 1 -
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제 2 장국내외기술개발현황 표 1. 2011 년국내복합제를이용한천연물신약개발현황 제품명주성분 ( 생약 ) 회사명개발형태적응증허가일자 신바로캡슐 시네츄라시럽 자오가, 우슬, 방풍, 두충, 구척, 흑두황련, 아이비엽 녹십자복합제골관절염 2011.1.25 안국약품복합제기관지염 2011.3.11 모티리톤정 현호색, 견우자 동아제약 복합제 기능성 소화불량증 2011.5.16-6 -
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Drug Name Company Mechanism Product Stage Administration prostaglandin F2 receptor agonist. Bimatoprost Allergan, Inc. stimulate the resting follicles to Phase II Topical growing follicle CB-03-01 Medicis Pharmaceutical Corporation steroid ester and a androgen antagonist Phase II Topical Refagro Kasiak Research Pvt. Ltd. autologous Human Platelet Lysate, stimulates new blood vessels to grow for the hair follicles Phase II Injection RK-023 R-Tech Ueno, Ltd. - Phase II Topical - 11 -
제 3 장연구개발수행내용및결과 - 12 -
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μ α α α α α α μ β β β β β β β - 15 -
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오가피 문주란 - 17 -
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2.55mg/g 이상 2.88mg/g 1.42mg/g 이상 1.62mg/g 1.31mg/g 이상 1.52mg/g - 22 -
가. 장기보존시험 120.0% 100.0% 80.0% 60.0% 40.0% Kaurenoic acid Lycorine Crinamine 20.0% 0.0% 초기 3 개월 6 개월 9 개월 12 개월 < 장기보존시험의지표성분함량변화 > 나. 가속시험 - 23 -
120.0% 100.0% 80.0% 60.0% 40.0% Kaurenoic acid Lycorine Crinamine 20.0% 0.0% 초기 1 개월 2 개월 3 개월 6 개월 12 개월 < 가속시험의지표성분함량변화 > - 24 -
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μ μ μ μ μ μ μ μ - 30 -
μ - 31 -
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μ μ μ α α α α - 33 -
α μ μ β α α α α μ α α - 34 -
α α α μ μ α α α α - 35 -
μ μ μ μ μ μ - 36 -
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μ - 38 -
β β β μ μ β β β β β β - 39 -
β β β β β β β β - 40 -
β β - 41 -
β β β β μ μ β β β β β β - 42 -
β β β β - 43 -
β β β - 44 -
β μ μ μ μ μ - 45 -
μ μ μ β β α β - 46 -
δ δ δ - 47 -
δ crinamine lycorine - 48 -
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HPLC 시스템 Shimadzu 20AD model 검출파장 290 nm (UV detector) 컬럼 Hector C18 (250 x 4.6 mm, 5 μm ) 컬럼온도 40 이동상 A : Water 100 % + 20mM ammonium acetate B : ACN 100 % 유속 1 ml/min 주입량 10 μl 시간 30 min Time B% Gradient 0.01 10 30 50 Lycorine standard Crinamine standard Sample ( 섬오가피 + 문주 란 ) - 51 -
Lycorine 농도 (mg/ml) R.T AREA 함량 (mg/g) Standard 0.24 10.4 4137000 Sample 10 10.2 272000 1.58 Crinamine 농도 (mg/ml) R.T AREA 함량 (mg/g) Standard 0.15 18.3 1368000 Sample 10 18.4 133000 1.46-52 -
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제 4 장목표달성도및관련분야에의기여도 a β - 64 -
전체항목연구개발세계최고결평가항목에서전단위수준 ( 주요성능 spec) 차지하는국내수준목표치과치비중 (%) 성능수준성능수준총 BHC ppm - - 0.2 불검출 1. 잔류농약총 DDT ppm 10 - - 0.1 불검출분석시험 Aldrin 외 ppm - - 0.01 불검출 달성도 (%) 100 평가방법 식약청고시외부의뢰 2. 중금속 함량분석 시험 납 ppm 5 5 5 비소 ppm 3 3 3 10 수은 ppm 0.2 0.2 0.2 카드뮴 ppm 0.3 0.3 0.3 100 식약청고시외부의뢰 3. 잔류이산화황 분석시험 ppm 10 30 30 30 0.7 100 식약청고시외부의뢰 4. 지표성분설정건 10 - - 2 2 100 KP 5. 대조약물 ( 미녹시딜 ) 대비효력 % 10 - - 100 94 100 양모제 효력평가시험 가이드라인 6. 제제의안정성시험 ( 장기보존, 가속시험 ) 건 10 적합여부적합여부 1 1 100 식약청고시 7. 단회투여독성시험 ( 설치류 ) 건 10 ICH 가이드라인에의거 GLP 규정에의거 1 1 100 의약품등의독성시험기준 (2009-116 호 ) 8. 4 주 DRF 시험건 10 ICH 가이드라인에의거 GLP 규정에의거 1 1 100 의약품등의독성시험기준 (2009-116 호 ) 9. 항원성시험건 10 ICH 가이드라인에의거 GLP 규정에의거 1 1 100 의약품등의독성시험기준 (2009-116 호 ) 10. 국소독성시험건 10 ICH 가이드라인에의거 GLP 규정에의거 1 1 100 의약품등의독성시험기준 (2009-116 호 ) - 65 -
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제 5 장연구개발성과및성과활용계획 - 67 -
Effects of dihydrotestoster one on rat dermal papilla cells in vitro Sang-Cheol Kim, Min-Kyoung Kim, Hye-JinBoo, Jung-Il Hee-Kyo Kang ung Kang Eun-Jl Kim, Jin-Won Hyun, Ji-Hoon Kang, Young-Sang Koh, Deok-Bae Park, Eun-Sook Yoo Pharmcol Res. 국외 SCI - 68 -
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2. 경제적성과 - 71 -
제 6 장연구시설 장비구매현황 - 72 -