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Synthesis of novel cyclodextrin derivatives and its application to search for the inhibitor of H.pyroli growth

新型环糊精衍生物的合成及其在寻找H.pyroli生长抑制剂中的应用

基本信息

批准号：
14560085
负责人：
HOJO Hironobu
金额：
$ 2.11万
依托单位：
TOKAI UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14560085/
关键词：
cyclodextrin Helicobacter Pyroli Inhibitor of growth 増殖阻害

项目摘要

Helicobacter pyroli is a gram-negative microaerobic bacterium, which has been proposed to be a cause of peptic ulcers. Laboratory growth of H.pyroli is carried out by using complex media containing serum, and blood derivatives. In stead, H.pyroli can be grown in media containing β-cyclodextrin. The reason of this fact is unknown yet, but it is suggested that CD traps the substances that is harmful for H.pyroli growth. If the trapped material can be identified, it will be a good lead compound for the development of the inhibitor of H.pyroli growth. From this point of view, we tried to isolate the CD-guest complex from the supernatant of the media used for the culture of H-pyroli. However, CD-guest complex had no UV absorption, causing its purification extremely difficult.In this research, we synthesized three β-CD derivatives, which will facilitate the purification of CD-guest complex. First compound is biotinylated β-CD. This CD is synthesized to purify the complex by avidin column. Ho … More wever, this compound was highly insoluble in water and thus, could not be used for the culture of H.pyroli. Next, we prepared β-CD carrying 4-bromo-2, 3, 5, 6-tetrafluorobenzoyl group. Bromine is composed of two isotopes, ^<79>Br and ^<81>Br with almost the same amount. Thus, elution fraction of this CD derivative will be easily identified by the characteristic mass spectrum derived from bromine. Although the culture of the H.pyroli using this CD was successful, the elution position of this CD derivative could not be identified by the mass spectrum. The last derivative, which carries fluorescein on β-CD, was also successfully used for the culture of H.pyroli. The post-cultural supernatant was separated by gel-filtration chromatography. The elution position of the CD derivative was clearly identified by its strong fluorescence and UV absorbance. The analysis of fraction containing the CD derivative by MALDI mass showed that the signal, which is about 100 atomic mass unit higher than the CD derivative was observed. This result might show that the CD derivative traps a substance, which has a molecular weight of about 100. We are now trying to identify its structure by ^1H-NMR.In conclusion, we succeeded in establishing a method to facilitate the purification of CD-guest compound from the cultural supernatant of H.pyroli, which will be useful to identify the growth inhibitor of H.pyroli. This method will be also applicable to find the inhibitor of other bacteria, such as B.pertussis, which can also be cultured in β-CD-containing media. Less

幽门螺杆菌是一种革兰氏阴性微需氧菌，已被认为是消化性溃疡的原因。通过使用含有血清和血液衍生物的复合培养基进行H.pyroli的实验室生长。相反，火烧菌可以在含有β-环糊精的培养基中生长。这一事实的原因尚不清楚，但它表明，CD陷阱的物质是有害的火烧菌生长。如果能对捕获的物质进行鉴定，它将是一个很好的先导化合物，用于开发抑制剂。从这个角度来看，我们试图从用于H-鹿蹄草培养的培养基的上清液中分离CD-客体复合物。本研究合成了三种β-CD衍生物，为CD-客体复合物的纯化提供了便利。第一种化合物是生物素化的β-CD。该CD被合成以通过亲和素柱纯化复合物。何 ...更多信息然而，该化合物高度不溶于水，因此，不能用于热解火杆菌的培养。接着，我们合成了带有4-溴-2，3，5，6-四氟苯甲酰基的β-环糊精。溴由两种同位素组成，^<79>Br和^<81>Br的含量几乎相同。因此，该CD衍生物的洗脱级分将容易通过源自溴的特征质谱鉴别。尽管使用该CD成功培养了火炎杆菌，但该CD衍生物的洗脱位置无法通过质谱识别。最后一种衍生物，其在β-CD上携带荧光素，也被成功地用于培养H. pyroli。通过凝胶过滤层析分离培养后上清液。CD衍生物的洗脱位置通过其强荧光和UV吸光度清楚地确定。通过MALDI质量对含有CD衍生物的级分的分析表明，观察到比CD衍生物高约100原子质量单位的信号。该结果可能表明CD衍生物捕获了分子量约为100的物质。本研究建立了一种从火炎菌培养上清中分离纯化CD-客体化合物的方法，为进一步鉴定火炎菌生长抑制剂奠定了基础。该方法也适用于寻找其他细菌的抑制剂，如B.pertussis，其也可以在含β-CD的培养基中培养。少