초록 ▼

화학적인 백혈병치료제는 심각한부작용 때문에 부작용이 적으면서도 내성이 없는 새로운 치료제를 천연물에서 개발하려는 시도가 진행되고 있는 것이 세계적인 추세이다. 이에 본 연구에서는 한의학적 약물배합 방법인 군신좌사법을 이용하여 한약재를 사용한 항백탕을 조제하여 백혈병 치료제 및 보조치료제 개발에 대한 연구를 시행하였다. 최신의 세포생물학적 분사생물학적 연구방법을 이용하여 연구를 수행한 결과 한약제에서 유래되는 선도물질들이 혈관신생 및 성장 억제나 bcr-abl protein kinase의 억제 및 항백혈병효능이 있었으며 이들을 군신좌

화학적인 백혈병치료제는 심각한부작용 때문에 부작용이 적으면서도 내성이 없는 새로운 치료제를 천연물에서 개발하려는 시도가 진행되고 있는 것이 세계적인 추세이다. 이에 본 연구에서는 한의학적 약물배합 방법인 군신좌사법을 이용하여 한약재를 사용한 항백탕을 조제하여 백혈병 치료제 및 보조치료제 개발에 대한 연구를 시행하였다. 최신의 세포생물학적 분사생물학적 연구방법을 이용하여 연구를 수행한 결과 한약제에서 유래되는 선도물질들이 혈관신생 및 성장 억제나 bcr-abl protein kinase의 억제 및 항백혈병효능이 있었으며 이들을 군신좌사법으로 배합한 항백탕 (I,II)의 인간 백혈병세포주인 HL-60, U-937, EL-4, K-562 등에 대한 세포고사, 세포분화에 대한 약리 효능 및 작용 기전을 규명하였고, 또한 EL-4를 이용한 in vivo에서도 그 효능을 입증하였다. 한편, balcalin imperatorin, 저근백피와 글리벡을 처리한 유전자 발현 변화와 항백탕(II) 복용 후 인간 혈중 백혈구 세포에서 유전자 발현 변화 등에 일정한 효과가 있음을 확인하였음.

Abstract ▼

I. Objective
Leukemia is an cancer of the white blood cells. In leukemia, a large number of white blood cells are produced in the stem cells of the bone marrow. These immature cells are incapable of normal differentiation and abnormally proliferated. Treatment for leukemia is complex. The primary

I. Objective
Leukemia is an cancer of the white blood cells. In leukemia, a large number of white blood cells are produced in the stem cells of the bone marrow. These immature cells are incapable of normal differentiation and abnormally proliferated. Treatment for leukemia is complex. The primary treatment for leukemla is chemotherapy by cytotoxic anticancer drugs. Differentiation therapy that may improve differentiation potential is also used in certain cases. However, leukemia and its treatment can cause a number of complications and side effects.
The objective of this project is to develop a therapeutic antileukemic composition of Oriental medicines which is based on the Oriental medicinal theory that can explain a possible interaction among major drugs in terms of Gun-Shin-Jawa-Sha, and its antileukemic mechanism.
II. Contents and Methods
1. Study of the effects of antileukemic agents and the apoptotic mechanism of herbal medicine
2. Development of differentiation inducing agents using herbal medicine and prescription which could compensate the side effect of all-trans retinoic acid- the rapid expression of tolerance and recurrence
3. Mechanism study of anticancer effects of Hangback-tang and development of Oriental medical prescriptions on the base of the mechanism of bcr-abl protein expression inhibition.
4. Elucidation of the genomic mechanism of Hangback-tang effect by the analysis of genome expression pattern using DNA-CHIP
III. Results of research and development
The herabl medicines to have the antileukemic effect were surveyed bibliographically and so we recognized that some herbal medicines and its chemical componets might have antileukemic effects, it was revealed that some of them, some agents derived from some herbal medicine had antileukemic effects. It was proved that the water extract of Ailanthus altissima had the inhibition effects of angiogenesis and the inhibition of bcr-abl tyrosine kinase activation. Also it was elucidated that Hangback-tang had the effect of apoptosis and differentiation on the cell line of HL-60 and U-937 and its mechanism and pharmacological reaction was researched. AS an in vivo assay, the antitumor effect on EL-4 was assayed. Derivatives of tetrahydorisoquinoline was assayed about the effects of apoptosis, differentiation induction, proliferation inhibition.
It was examined the induction of HL-60 cell differentiation by treatment of the water extract of Rhizoma Zingiberis Siccatum. The level of inhibition was identical to those of ATRA-treated cells. Rhizoma Zingiberis Siccatum extract treatment caused a significant change in NBT reduction (47%) and enhanced ATRA-induced NBT reduction. Treatment of Rhizoma Zingiberis Siccatum to HL-60 cells increased only CD11b expression in the cells, and also increased markedly G0/G1 stage arrest. This can suggest that Rhizoma Zingiberis Siccatum induced the differentiation of HL-60 cell and enhanced ATRA-induced differentiation predominantly along the granulocytic lineage. The results present here show that Rhizoma Zingiberis Siccatum extract contains the potential effect of differentiation induction or acts as a agent enhancing the induction of differentiation.tiation of HL-60 cells in Hwoangbaectang composition dependent manner.
Baicalin, a biologically active flavonoid form the roots of Scutallaria baicalensis (Skullcap), have been reported to not only function as anti-oxidants but also cause anticancer effect. We investigated the mechanism of baicalin-induced cytotoxicity and the macro scale gene expression analysis in leukemia cell line, HL-60 cells. Baicalin (10 ${\mu}M$) were used to treat the cells for 6h, 12h, 24h, 48h and 72h. In a human cDNAchip study of 65,000 genes evaluated 6, 12, 24, 48, 72 hours after treated with Baicalin in HL-60 cells. Hierarchical cluster against the genes which showed expression changes by more than two fold. One hundred one genes were grouped into 6 clusters according to their profile of expression by a hierarchical clustering algorithm. For genes differentially expressed in response to baicalin treatment, we tested functional classes based on Gene Ontology (GO) terms. This study provides the most comprehensive available survey of gene expression changes in response to baicalin treatment in HL-60 cell line.
Imperatorin, a biologically active furanocoumarin from the roots of Angelica dahurica (Umbelliferae), was mutagenic and induced transformation of mouse fibroblast cell lines, whereas it provided inhibiting effects on mutagenesis and carcinogenesis induced by various carcinogens. We investigated the macro scale gene expression analysis on the HL-60 cells treated with imperatorin. Imperatorin (10 ${\mu}M$) were used to treat the cells for 6h, 12h, 24h, 48h, and 72h. In a human cDNAchip study of 10,000 genes evaluated 6, 12, 24, 48, 72 hours after treated with imperatorin in HL-60 cells. Hierarchical cluster against the genes which showed expression changes by more than 2 fold. Three hundred eighty six genes were grouped into 6 clusters by a hierarchical clustering algorithm. Pathway analysis using gene microarray pathway profiler that is a computer application designed to visualize gene expression data on screen representing biological pathways and groupings of genes.

목차 Contents

• 제 1 장 연구개발과제의 개요...73
• 제 2 장 국내외 기술개발 현황...78
• 제 3 장 연구개발수행 내용 및 결과...80
• 제 4 장 목표달성도 및 관련분야에의 기여도...95
• 제 5 장 연구개발결과의 활용계획...97
• 제 6 장 연구개발과정에서 수집한 해외과학기술정보...97
• 제 7 장 참고문헌...97
• 제 8 장 첨부서류...97
• 제 1 세부과제명 : 항백탕을 이용한 백혈병의 세포고사 유도 치료법 개발...309
• 제 2 세부과제명 : 급성 백혈병치료를 위한 세포분화 유도법 개발...331
• 제 3 세부과제명 : 항백탕가미방의 protein kinase 기능조절에 의한 항백혈병 효능 연구...348
• 제 4 세부과제명 : DNA chip을 이용한 항백탕의 항백혈병 특정 연구...371