초록 ▼

당해 연도 본 연구진은 1차연도 연구를 바탕으로 oxythiamine과 다른 화합물들과의 병용 처리시 상승되는 항녹농균 효과를 체계적으로 검증함과 동시에, oxythiamine의 활성기작에 있어 thiamine monophosphate kinase의 역할을 규명하여, 새로운 표적으로써 thiamine 대사기작의 활용성을 제시함.
또한 대표적인 그람양성 슈퍼박테리아인 폐렴구균과 황색포도상구균에 대한 연구를 통하여 새로운 항폐렴구균 약재검색 시스템을 개발, 검증하였고, 항생제에 의해 유도되는 황색포도상구균 특이적 생리반응을 분자

당해 연도 본 연구진은 1차연도 연구를 바탕으로 oxythiamine과 다른 화합물들과의 병용 처리시 상승되는 항녹농균 효과를 체계적으로 검증함과 동시에, oxythiamine의 활성기작에 있어 thiamine monophosphate kinase의 역할을 규명하여, 새로운 표적으로써 thiamine 대사기작의 활용성을 제시함.
또한 대표적인 그람양성 슈퍼박테리아인 폐렴구균과 황색포도상구균에 대한 연구를 통하여 새로운 항폐렴구균 약재검색 시스템을 개발, 검증하였고, 항생제에 의해 유도되는 황색포도상구균 특이적 생리반응을 분자생물학적 측면에서 확인하여 새로운 항균물질 발굴 및 항생제내성 기작연구의 근간을 마련함.

( 출처 : 보고서 요약서 3p )

Abstract ▼

제1절 슈퍼박테리아
Antibiotic resistance is rapidly spreading among bacteria, such as Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pneumoniae. Infections caused by these superbugs are often life-threatening and extremely difficult to treat due to resistance to antibiotics.
Althoug

제1절 슈퍼박테리아
Antibiotic resistance is rapidly spreading among bacteria, such as Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus pneumoniae. Infections caused by these superbugs are often life-threatening and extremely difficult to treat due to resistance to antibiotics.
Although new antibiotics are urgently needed, very few new drugs are currently in development. In order to discover novel antibacterial drugs, we previously developed a new culture broth medium based on artificial lung-mucus medium to replicate a condition relevant for pseudomonas infection in vivo. A few hits compounds including oxythiamine were identified from a screen of small molecules against Pseudomonas aeruginosa. This year we continue to investigate oxythiamine in order to confirm its mechanism and characteristics. Mutant and biochemical studies suggested that the phosphorylation occurs through bacterial thiamine biosynthetic pathway especially through thiamine monophosphate kinase (ThiL). In addition, combination assays of oxythiamine with structurally distinct 8 different compounds demonstrated that oxythiamine can create synergistic antipseudomonal effects with many of them. A new assay for S. pneumoniae was developed to discover antipneumococcal molecules. The developed assay was applicable for different S. pneumoniae strains including a reference strain and multidrug resistant strains. Using the assay, screenings with about 7800 compounds in FDA & bioactive libraries are performed validating the assay. In order to decipher the mechanism(s) leading to antibiotic resistance, we investigated transcriptomic changes in S. aureus exposed to different antibiotics. From results of these studies, we identified a hypothetical protein (SA-13280) that is induced at early time point (60 minutes) in a concentration-dependent manner. Based on this finding, we developed a new assay for S. aureus using GFP reporter system controlaed by SA_13280 promoter.

( 출처 : SUMMARY 6p )

제2절 리슈만질환
Leishmaniasis is a vector borne parasitic disease responsible for serious socioeconomics and public health problems in developing and undeveloped countries. Particularly the disease is spread out through Latin America, Africa Asia as well as southern Europe with 16 million cases and 30,000 deaths annually. Currently, there is no available vaccines and the treatments such as antimonials, Amphotericin B, Miltefosine and Paramomycin are ineffective with high adverse effects limiting the useage of these drug. Therefore, there is an urgent need to develop safe and effective drugs against Leishmaniasis. Through this project, we aim to discover antileishmanial lead compound utilizing cell based drug discovery approach.
The primary aim of this project is to develop intracellular Leishmania assay and adapt it to high throughput format to screen for hit compounds for further development. Upon identification of hits, the compounds will be subjected to secondary assay to monitor the effects on mitochondria of the parasite. The mitochondria specific compounds will then be optimized for lead discovery as well as mode of action studies on the mitochondria of the parasite. In the later phase of the project, we will perform in vivo Leishmania efficacy test as a prove of concept study.
During the second year of the project, we have successfully screened two sets of libraries and developed secondary assays related to kinetoplast replication for identifying inhibitors against the organelle. For the screening, we have first screened 1,742 bioactive compounds in duplicate resulting in R2 of 0.92 and identified 23 hit compounds with >50% inhibitions with <50% toxicity. Based on EC50 values, the most potent class of compounds were found to be mTOR Kinase inhibitors which should the potential for drug repositioning. For 50,000 compounds screening, the overall Z’was 0.8 and 154 hits were identified. Based on compound clustering, 17 clusters and 46 singtons were identified. Representative hits form each clusters were repurchased and EC50 values were determined. Based on the activity, we will further optimize selected cluster. For the last part of the project, we have successfully developed click-chemistry based replication assay which can distinguish the phonotypes with complete replication of kinetoplast and nucleus or not. Also we expressed , purified and developed assay for hetero-dimeric Leishmania Topoisomerase I assay. Using the assay, we have exploit the mechanism of action of current treatment pentamidine and identified new diamidine type of antileishmanial compounds which selectively inhibits the replication process of kinetoplast.

( 출처 : SUMMARY 7p )

목차 Contents

• 표지 ... 1
• 제 출 문 ... 2
• 보고서 요약서 ... 3
• 요 약 문 ... 4
• SUMMARY ... 6
• CONTENTS ... 8
• 목차 ... 9
• 제1장 연구개발 과제의 개요 ... 10
• 제1절 슈퍼박테리아 ... 10
• 제2절 리슈만 질환 ... 10
• 제2장 국내외 기술개발 현황 ... 11
• 제1절 슈퍼박테리아 ... 11
• 제2절 리슈만 질환 ... 12
• 제3장 연구개발 수행 내용 및 결과 ... 14
• 제1절 슈퍼박테리아 ... 14
• 제2절 리슈만 질환 ... 23
• 제4장 목표달성도 및 관련분야의 기여도 ... 31
• 제5장 연구개발결과의 활용계획 ... 39
• 제1절 슈퍼박테리아 ... 39
• 제2절 리슈만 질환 ... 39
• 제6장 연구개발 과정에서 수집한 해외과학기술정보 ... 40
• 제7장 연구시설·장비 현황 ... 41
• 제8장 참고문헌 ... 48
• 끝페이지 ... 49