초록 ▼

Ⅳ. 연구개발결과
□ 용량별 혈당강하 효능시험
당뇨유발모델인 db/db 마우스(수컷, 7 주령)에 시험물질 누에추출물분말을 사료에 섞어 4 주 반복급이 하였을 때 혈당강하 효력을 알아보고자 수행하였다.
○ 혈당 측정결과, 시험물질 누에추출물분말 22.5 mg/kg/day군에서 1-4 주에 부형제 대조군에 비해 통계학적으로 유의한 혈당 강하가 관찰되었다.
○ 사료섭취량 측정결과, 시험물질투여군에서 부형제대조군에 비해 통계학적으로 유의한 변화는 없었다.
○ 물섭취량 측정결과, 시험물질투여군에서 부형제대조군에

Ⅳ. 연구개발결과
□ 용량별 혈당강하 효능시험
당뇨유발모델인 db/db 마우스(수컷, 7 주령)에 시험물질 누에추출물분말을 사료에 섞어 4 주 반복급이 하였을 때 혈당강하 효력을 알아보고자 수행하였다.
○ 혈당 측정결과, 시험물질 누에추출물분말 22.5 mg/kg/day군에서 1-4 주에 부형제 대조군에 비해 통계학적으로 유의한 혈당 강하가 관찰되었다.
○ 사료섭취량 측정결과, 시험물질투여군에서 부형제대조군에 비해 통계학적으로 유의한 변화는 없었다.
○ 물섭취량 측정결과, 시험물질투여군에서 부형제대조군에 비해 통계학적으로 유의한 변화는 없었다.
○ 혈액생화학적 검사결과, AST, ALT, TCHO, TG, LDL 및 HDL에서 시험물질의 영향으로 판단되는 혈액생화학적인 변화는 관찰되지 않았다. GLU에서는 시험물질 22.5 mg/kg/day군에서 부형제대조군에 비해 통계학적으로 유의한 감소가 관찰되었다.
□ 누에추출물 분말의 열 안정성 시험
○ 1-DNJ함량에서는 무처리, 60℃, 100℃까지는 큰 변화가 없었으나 150℃처리구에서 급격하게 감소하였다.
○ 당뇨모델 마우스에서의 혈당강하 효능시험에서도 150℃처리구에서만 거의 효과가 없었다.
○ 누에추출물분말의 멸균처리가 필요한 경우 100℃/30분을 초과해서는 안될 것으로 판단된다.
□ 누에추출물분말의 안전성 독성연구
○ 설치류 단회 경구투여 독성시험 : 개략의 치사량은 암수 5000 mg/kg 상회
○ 설치류 4 주간 반복 경구투여 DRF 독성시험 : 최고농도 2000 mg/kg 추천
○ 비설치류(Beagle dog)에 DE (dose escalation) 법을 이용한 단회 경구투여 독성시험 : 최대내성용량은 암수 2000 mg/kg
○ 복귀돌연변이시험 : 음성
○ 염색체 이상시험 : 음성
○ 소핵시험 : 음성
○ 조제물 시험: 멸균주사용수 현탁액 중 누에추출물분말의 HPLC를 이용한 분석법 검증 : 균질 및 냉장 보관 조건에서 7 일간 안정
○ 설치류 13 주간 반복 경구투여 독성시험 및 4 주 회복시험 : 무독성량: 암수 2000 mg/kg, 표적장기: 없음
○ 안전성 약리- 비글을 이용한 Telemetry 및 CHO cell을 이용한 허그 assay : 이상 없음
○ 안전성약리 - 랫드를 이용한 단회 경구 투여후 호흡수 및 호흡량 측정시험 : 이상 없음
○ 안전성 약리- 마우스를 이용한 체온 및 전신행동 시험 : 이상 없음

Abstract ▼

We investigated the 1-DNJ contents of silkworm larvae extracts according to the developmental stages and three silkworm varieties. And we compared with polyhydroxylated alkaloid contents such as 1-deoxynojirimycin(DNJ), Fagomine, and 1,4-dideoxy-1,4-imino-Darabinitol (DAB) of silkworm extracts. . In

We investigated the 1-DNJ contents of silkworm larvae extracts according to the developmental stages and three silkworm varieties. And we compared with polyhydroxylated alkaloid contents such as 1-deoxynojirimycin(DNJ), Fagomine, and 1,4-dideoxy-1,4-imino-Darabinitol (DAB) of silkworm extracts. . In addition, we evaluated the glucose lowering effectof silkworm extracts powder in db/db mice.
The contents of 1-DNJ was the highest with 18.4mg/dried 100g and 18.3mg/dried 100g in Yeonnokjam 5^thinstar3^rddays and Hansaengjam 5^thinstar3^rddays. The larvaeof 5^thinstar3^rddays showed higher contents of 1-DNJ than the larvae 5thinstar5^thdays among all varieties.The glucose lowering effects of the silkworm extracts and powder of Yeonnokjam are tested on the db/db mice. The blood glucose level decreased significantly in YR70 group.
The silkworm extracts and acarbose were administered orally with 180mg/kg, 90mg/kg, 45mg/kg and 22.5mg/kg of silkworm extracts and 50mg/kg of acarbose for 4 weeks. Water intake were not significant changes statistically when compared between control group and silkworm extracts treated group. Silkworm extract powder 22.5 mg/kg/day group compared with the control group after administered 4 weeks, a statistically significant decreased the blood glucose level. Blood biochemical changes of the AST, ALT, TCHO, TG, LDL and HDL did not observed. But blood biochemical changes of the GLU decreased statistically significant in 22.5 mg / kg / day group compared to the control group. The epididymal fat weight of silkworm extract powder treated were decreased significantly observed in 22.5mg/kg and 180mg / kg / day group compared with the control group. Perirenal fat weight, however, there were no significant changes in the statistical. Based on these results, it is considered that the silkworm extracts has favorable effect to inhibit the change on the blood glucose levels in diabetes model mice.
Silkworm powder containing 1-deoxynojirimycin (DNJ) is promising as a complementary and alternative medicine (CAM) agent in Korea. Silkworm powder produced Yeonnokjam 5thinstar3rddays in National Academy Agricultural Science. The powder was derivatized with 9-fluorenylmethyl chloroformate (FMOC-Cl), and DNJ-FMOC content was measured by HPLC. We investigated the 1-DNJ contents of silkworm powder according to the change temperature. The glucose lowering effects of the silkworm powder treated changing temperature. The contents of 1-DNJ was the lowest with T150(150℃/30min). Another treatments were very stable
The silkworm powder were administered orally with 20mg/kg/day for 4 weeks.
Water intake were not significant changes statistically when compared between control group and treated group. T0 (Control)and T60(60℃/30min) group compared with the control group after administered 4 weeks, a statistically significant decreased the blood glucose level but T100(100℃/30min) and T150(150℃/30min ) was not decreased.Blood biochemical changes of the TG were significant decreased statistically compared with the control group.
Based on these results, it is considered that the silkworm powder was stable upon heating to 100℃ for 30min but heated silkworm powder at 150 ℃ for 30min was unstable.
The objective of this toxicity study is to evaluate the toxicity of the Silkworm powder for its utilization in the field of medicines. The approximate lethal dose (ALD) was considered to be 5,000 mg/kg in males and females in a single oral dose test and the no observed adverse effect level (NOAEL) was 2,000 mg/kg/day and the target organ was not identified in a 13-week repeated oral dose toxicity study with a 4-week recovery period in Sprague-Dawley rats.
The maximum tolerance dose (MTD) was considered to be 2,000 mg/kg/day in male and female in a single oral dose study by using the dose escalating method in Beagle dogs.
The test article was considered to be negative in the bacterial reverse mutation assay, micronuclei assay in the bone marrow cells of ICR mice and in the chromosomal aberration assay using CHL cells. It was considered to be suitable that the quantitative method using HPLC was applied for the analysis of 1-deoxynojirimycin in the Silkworm powder in sterile water for injection.
The dosing formulation was homogeneous at doses of 5~200 mg/mL of the test article and stable for 7 days under the refrigerated storage conditions(3.46 ± 0.22 ℃).
Further, It was concluded that this Silkworm powder did not effect in a safety pharmacology study on body temperature and general behavior in mice, on the respiration rate and volume of Sprague-Dawley rats, on the telemetry in Beagle dog and in the herg assay using CHO cells.

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 요약문 ... 3
• SUMMARY ... 7
• 목차 ... 9
• 제 1 장 서 론 ... 10
• 제 2 장 국내외 기술개발 현황 ... 11
• (1) 국내 연구 현황 ... 11
• (2) 국외 연구 현황 ... 11
• 제 3 장 연구개발수행 내용 및 결과 ... 11
• Ⅰ절 재료 및 방법 ... 11
• Ⅱ절 결과 ... 24
• Ⅲ절 적요 ... 98
• 제 4 장 연구개발목표 달성도 및 대외기여도 ... 100
• 1절 연구목표대비 달성도 ... 100
• 2절 정량적 성과 ... 100
• 제 5 장 연구개발결과의 활용계획 ... 103
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보 ... 103
• 제 7 장 기타 중요 변동사항 ... 103
• 제 8 장 국가과학기술종합정보시스템에 등록한 연구장비 현황 ... 103
• 제 9 장 참고문헌 ... 104
• 끝페이지 ... 108