초록 ▼

○ 연구결과
분자 설계 기법을 통하여 제시된 Tyrosine kinase 저해제를 제조하기 위하여， 용액상 및 고체상 모두에서 분자조합 기술을 개발하여， 하이드라진， qunazoline, aromatic amino acid, histamine, diagonal piperidine 라이브러리 및 spiro-compòund등의 다양한 pharmacophore를 포함하는 각각의 라이브러리를 각각 48 ~ 650종씩 제조하였다.
제조된 라이브러리에 대한 약효검색으로써 tyrosine kinase assay와 MTT assay를

○ 연구결과
분자 설계 기법을 통하여 제시된 Tyrosine kinase 저해제를 제조하기 위하여， 용액상 및 고체상 모두에서 분자조합 기술을 개발하여， 하이드라진， qunazoline, aromatic amino acid, histamine, diagonal piperidine 라이브러리 및 spiro-compòund등의 다양한 pharmacophore를 포함하는 각각의 라이브러리를 각각 48 ~ 650종씩 제조하였다.
제조된 라이브러리에 대한 약효검색으로써 tyrosine kinase assay와 MTT assay를 확립 및 실시하여 선도물질을 도출하였고， 이를 근거로한 3차원적 분자구조활성 상관관계 연구(3D-QSAR)를 통하여 새로 제시되는 분자구조를 대상으로 2차 라이브러리를 제조하는 선도물질 최적화 연구를 수행하였다.
선도물질 최적화 연구로써， 제조된 2차 라이브러리 중에서 in vitro 약효검색을 통하여 선정된 우수물질 (YH3411， YH3475, YH3476)들에 대한， i) nude mice assay를 실시하여 in vivo에서 항암효과를 평가하였고， ii) 체내동태 평가를 실시하여 시험물질의 체내동태의 특성과 경구투여시의 생체이용율을 구하였고， iii) 독성연구를 수행하여 시험물질의 안정성 검토를 통하여 신약후보물질로서의 개발가능성을 모색하였다.

Abstract ▼

III. Result
A general and efficient method for the solid phase synthesis of aza-peptide mimetic library incorporating hydrazine residues as a modified amino acid backbone was investigated and developed starting from hydroxymethyl polystyrene using a Fmoc-hydrazine scaffold. A 650 compound library

III. Result
A general and efficient method for the solid phase synthesis of aza-peptide mimetic library incorporating hydrazine residues as a modified amino acid backbone was investigated and developed starting from hydroxymethyl polystyrene using a Fmoc-hydrazine scaffold. A 650 compound library has been constructed over 7 steps in good yields with high purity using a diverse set of commercially available amino acid, primary and secondary amine, acyl chloride, and iso(thio)cyanate building blocks.
Also, a general and efficient method for the solution phase synthesis of qunizoline library was investigated and developed using several halogenated quinazolines scaffolds. A 250 compound library has been constructed over 5 steps in good yields with high purity using a diverse set of commercially available acid, primary and secondary amine, acyl chloride building blocks.
Design and synthesis of aromatic ammo acid, diamine, and histamine libraries were performed. Diphenylsilydiethylene-(DPSide-) group has been developed as a new primary amine protecting group. The new protecting group exhibits novel orthogonality against other commonly used amine-protecting groups and excellent chemoselectivity for unbranched a -amino acid esters. Removal of this protecting group was effective under mild fluoride-based cleavage conditions.
We have developed diagonal combinatorial library technique based on the indexed library system. The new system was compared with the known indexed system using statistical analysis and the result indicated that the new system would have more chance to show hit The real test using 10x10 matirx compounds showed that the diagonal combinatorial library was better or complimentary to the indexed library at least. Using reductive amination reaction and salt forming strategy, a successful methodology to generate solution phase combinatorial libraries has been developed. We were able to produce 10x10 size library and the methodology could easily generate 20x20 size library in practice without applying special tools or technologies. To support generating diverse combinatorial libraries, we also developed a synthetic method to construct spirocyclic amine compounds with structural diversity.
For the discovery of the potent EGFr tyrosine kinase inhibitor, their inhibitory activity was evaluated through ELISA tyrosine kinase assay using partially purified EGFr tyrosine kinase purified from membrane fractions of human epidermoid carcinoma A431 cells harboring overexpressed EGFr. The inhibitory activity of compounds on growth of human A431 cell was examined using MTT assay. Many compounds including YH3441, YH3475, and YH3476 were identified as the potent inhibitors on the phosphorylation by human EGFr tyrosine kinase with IC₅₀s of nanomolar range. Through the intensive optimization, it indicates that YH3475 is the most potent tyrosine kinase inhibitor among those compounds leading to the loss of EGFr-driven malignancy. After intravenous administration (10 mg/kg) of three lead compounds (YH3441, YH3475, and YH3476) to mouse, the plasma concentrations showed monoexponential decay, and t_l/2 values were below 10 min, respectively. After oral administration of these compounds (25 mg/kg) , the AUC values were observed. The oral bioavailability were relatively low range (10-20%) at the studied dose.
Acute toxicity of three lead compounds administered orally with single dose was conducted in mice. The LD₅₀ values of YH3441 and YH3475 were above 5,000 mg/kg in male mice, respectively.

목차 Contents

• 표지 ... 1
• 제출문 ... 3
• 요약문 ... 4
• 영문요약문(SUMMARY) ... 11
• 목차 ... 13
• 제1장 서론 ... 15
• 제1절 연구개발의 목적 ... 16
• 제2절 연구개발의 필요성 ... 16
• 제3절 연구개발의 범위 ... 20
• 제2장 국내외 기술개발 현황 ... 22
• 제1절 국외현황 ... 23
• 제2절 국내현황 ... 30
• 제3절 연구결과의 국내외 현황에서의 위치 ... 32
• 제3장 연구개발 수행 내용 및 결과(제1세부과제) ... 33
• 제1 세부과제: 약효검색을 통한 신규 항암제의 도출 (유한양행) ... 34
• 제2 세부과제: Small Organic Molecule Library의 개발 (서울대) ... 99
• 제3 세부과제: Diagonal Combinatorial Library의 개발 (KAIST) ... 121
• 제4장 연구개발 달성도 및 대외기여도 ... 144
• 제1절 연구개발 목표 달성도 ... 145
• 제2절 관련분야의 기술발전에의 기여도 ... 149
• 제5장 연구개발결과의 활용성과 및 계획 ... 150
• 제1절 연구개발결과의 활용성과 ... 151
• 제2절 연구개발결과의 활용계획 ... 152
• 제6장 기타 주요변경사항 ... 152
• 제7장 참고 문헌 ... 153
• 최종보고서 요약(초록) ... 159
• 끝페이지 ... 159