香豆素類化合物與乙酰膽堿酯酶的相互作用研究
發(fā)布時間：2019-05-15 11:06
【摘要】：阿爾茨海默病是引起中老年人癡呆最常見的疾病。乙酰膽堿酯酶（AChE）抑制劑是當(dāng)前治療該疾病的主要手段。目前臨床使用的AChE抑制劑存在生物利用率低、活性不是特別高、選擇性低、具有不同程度的毒性及副作用等缺點(diǎn)，尋求新的高效低毒的AChE抑制劑是藥物化學(xué)家們研究的熱點(diǎn)之一。香豆素類化合物是具有苯并吡喃環(huán)母核的一類天然化合物，研究表明，該類化合物具有一定的AChE抑制活性，具有潛在的藥用價值。藥物發(fā)揮藥效的化學(xué)本質(zhì)是藥物分子與生物靶分子之間的相互作用，藥物藥效的高低與其和靶分子之間的相互作用密切相關(guān)，因此在分子水平上研究香豆素類化合物與AChE的相互作用，對于探討該類化合物抑制AChE活性的分子機(jī)理及新的AChE抑制活性化合物的篩選和研究都具有重要意義。香豆素類化合物的生物活性與其苯并吡喃環(huán)上取代基的種類和位置密切相關(guān)，這些取代基團(tuán)如何影響該類化合物與AChE的相互作用，以及二者之間的體外相互作用與其對酶活性的抑制之間的相關(guān)性如何，都是十分值得研究的問題，然而，目前尚未見相關(guān)報導(dǎo)。 針對上述問題，本論文選擇了結(jié)構(gòu)取代不同的13種香豆素類化合物，通過使用多種光譜方法并結(jié)合分子模擬技術(shù)研究了上述化合物與AChE之間的相互作用，探討了該類化合物與AChE相互作用的化學(xué)特征和空間結(jié)合模式；比較了不同結(jié)構(gòu)取代對該類化合物與AChE相互作用的影響；利用Ellman法對AChE的酶活性進(jìn)行研究，探討了不同結(jié)構(gòu)取代對該類化合物對AChE酶活的抑制影響。本研究所獲結(jié)果對于尋找該類化合物發(fā)揮藥效的活性基團(tuán)以及篩選新型的香豆素類AChE抑制劑具有理論的參考意義。具體內(nèi)容如下： 1香豆素類化合物與AChE相互作用的光譜法研究：利用熒光、紫外-可見吸收光譜法研究了13種香豆素類化合物與AChE的相互作用，探討了其相互作用的機(jī)理，并對相互作用中的結(jié)合常數(shù)、結(jié)合位點(diǎn)數(shù)以及二者的結(jié)合距離作了計算。由結(jié)果可知，該類化合物對AChE的內(nèi)源熒光具有較強(qiáng)的猝滅作用，猝滅的機(jī)制主要為形成復(fù)合物的靜態(tài)猝滅，且能引起AChE構(gòu)象的變化，疏水作用力和范德華力為其與AChE間相互作用的主要作用力，同時還存在氫鍵及靜電作用力；該類化合物與AChE間的結(jié)合距離均小于7nm；不同結(jié)構(gòu)取代化合物相互作用大小的比較表明：香豆素母核4位上羥基的取代以及7位上甲氧基和乙氧基的取代對增強(qiáng)該類化合物與AChE的相互作用具有重要影響。 2香豆素類化合物與AChE相互作用的分子模擬研究：以電鰩乙酰膽堿酯酶的三維空間結(jié)構(gòu)作為模板蛋白，用Autodock4.0軟件對AChE與所研究的香豆素類分子進(jìn)行分子對接，主要對二者相互作用的相互作用能，結(jié)合部位及作用力類型進(jìn)行了判斷，結(jié)果表明各香豆素化合物均能與AChE的活性位點(diǎn)發(fā)生相互作用，且與AChE作用的氨基酸主要為疏水性以及帶電荷的氨基酸，在香豆素類分子與AChE形成的復(fù)合物中，范德華力及疏水作用力占主導(dǎo)，同時存在氫鍵與靜電作用力，在分子對接研究中對相互作用中作用力類型的判斷與熒光方法基本一致；由于香豆素類化合物中香豆素環(huán)的存在，其可能與AChE殘基產(chǎn)生π-π作用，，π-π共軛也可能是導(dǎo)致AChE熒光猝滅的原因。各個香豆素類化合物由于取代基結(jié)構(gòu)、位置或者個數(shù)不同，導(dǎo)致其與AChE相互作用的氨基酸殘基種類和個數(shù)也不同，與殘基形成的氫鍵個數(shù)也有區(qū)別，實驗結(jié)果表明4位上羥基的取代形成的氫鍵較多。 3香豆素類化合物對AChE活性的改變研究：以碘化硫代乙酰膽堿為底物，利用Ellman法定量地評價了AChE的活性，通過各個化合物對AChE活性的半數(shù)抑制濃度的計算來比較各個香豆素類化合物對酶抑制能力的大小。結(jié)果表明該類化合物對AChE活性均有一定的抑制作用，香豆素母核4，7位上的取代顯示了較好的AChE抑制活性，4，7位上羥基的取代以及羥基取代個數(shù)的增加使香豆素類化合物對AChE抑制活性的增強(qiáng)尤為明顯。
[Abstract]:Alzheimer's disease is the most common disease in the middle-aged and the elderly. Ethylcholine esterase (AChiE) inhibitors are the main means for the current treatment of the disease. The AChiE inhibitor currently used in clinical use has the disadvantages of low bioavailability, high activity, low selectivity, different degree of toxicity and side effect, and the like, and the novel high-efficiency and low-toxicity AChiE inhibitor is one of the hot spots of the drug chemists. Coumarin compounds are a kind of natural compound with benzo-benzene ring mother nucleus, and the study shows that the compound has certain AChiE inhibitory activity and has potential medicinal value. The chemical nature of the drug effect is the interaction between the drug molecules and the biological target molecules, the drug effect is closely related to the interaction between the drug molecules and the target molecules, It is of great significance to study the molecular mechanism of the compound to inhibit the activity of AChE and the screening and study of the new AChE inhibitory active compound. the biological activity of the coumarin compound is closely related to the species and the position of the substituents on the benzo-homopolar ring, how these substituents affect the interaction of the compound with the achaE, and how the in vitro interaction between the two is related to its inhibition of the enzymatic activity, It is a matter of great value to study, however, no relevant reports have been reported at this time. In the light of the above-mentioned problems, the structure is selected to replace 13 kinds of coumarin compounds, and the interaction between the above-mentioned compounds and the AChE is studied by using a variety of spectral methods and the molecular simulation technology. In this paper, the chemical and spatial-binding modes of the interaction between the compounds and the AChE are discussed. The effect of the substitution of the different structures on the interaction of the compounds with the AChE is compared. The enzyme activity of the AChE is studied by Elman's method. The effect of substitution of different structures on the activity of AChiE in this kind of compound was studied. In response, the results of this study are useful for finding the active groups of these compounds and to screen new type of coumarin AChiE inhibitors. I. Specific contents such as In this paper, the interaction of 13 kinds of coumarin compounds with AChE was studied by fluorescence and ultraviolet-visible absorption spectrometry, and the interaction mechanism was discussed. the combination constant, the combination of the number of bits, and the combined distance of the two The results show that the compounds have a strong quenching effect on the endogenous fluorescence of AChE. The quenching mechanism is mainly to form the static quenching of the complex, and can cause the change of the conformation of the AChE, the hydrophobic force and the van der Waals force are the main factors to interact with the AChE. the interaction distance between the compound and the AChE is less than 7 nm; the ratio of the interaction size of the different structures to the compound It is shown that the substitution of the hydroxyl groups in the 4-position of the coumarin and the substitution of the methoxy and ethoxy groups on the 7-position can be used to enhance the interaction of the compound with the AChE. A molecular simulation study on the interaction of 2-coumarines and AChE was carried out. The three-dimensional space structure of Choline esterase was used as a template protein, and the interaction of AChE with the studied coumarin-based molecules was carried out by Audiock4.0 software. The interaction energy, the binding site and the type of force are determined. The results show that the coumarin compounds can interact with the active sites of the AChiE, and the amino acids that interact with the AChiE are mainly hydrophobic and charged amino acids, which are formed in the coumarin-like molecules and the AChiE. In the complex, the van der Waals force and the hydrophobic force are dominant, and the hydrogen bond and the static force are present at the same time, and the judgment of the force type in the interaction is basically the same as that of the fluorescence method in the molecular docking study; and the coumarins in the coumarin compound The presence of a prime ring, which may be the same as that of the AChE residue, may also be the result of the AChE fluorescence The reason of the quenching is that the number of amino acid residues which can interact with the AChE is different due to the different structure, position or number of the substituents, and the number of hydrogen bonds formed by the residues is different, and the experimental results show that the substitution of the hydroxyl in the 4-position is formed. The changes of the activity of the 3-coumarin compound to the activity of AChE were studied. The activity of AChE was evaluated by Ellman, and the activity of AChE was evaluated by Ellman. The results show that the compound has a certain inhibitory effect on the activity of AChE, the substitution of the 4 and 7 positions of the coumarin shows better AChiE inhibitory activity, the substitution of the hydroxyl groups at the 7-position and the increase of the number of the hydroxyl groups make the coumarin compounds inhibit the AChE.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R969.2
本文編號：2477455
[Abstract]:Alzheimer's disease is the most common disease in the middle-aged and the elderly. Ethylcholine esterase (AChiE) inhibitors are the main means for the current treatment of the disease. The AChiE inhibitor currently used in clinical use has the disadvantages of low bioavailability, high activity, low selectivity, different degree of toxicity and side effect, and the like, and the novel high-efficiency and low-toxicity AChiE inhibitor is one of the hot spots of the drug chemists. Coumarin compounds are a kind of natural compound with benzo-benzene ring mother nucleus, and the study shows that the compound has certain AChiE inhibitory activity and has potential medicinal value. The chemical nature of the drug effect is the interaction between the drug molecules and the biological target molecules, the drug effect is closely related to the interaction between the drug molecules and the target molecules, It is of great significance to study the molecular mechanism of the compound to inhibit the activity of AChE and the screening and study of the new AChE inhibitory active compound. the biological activity of the coumarin compound is closely related to the species and the position of the substituents on the benzo-homopolar ring, how these substituents affect the interaction of the compound with the achaE, and how the in vitro interaction between the two is related to its inhibition of the enzymatic activity, It is a matter of great value to study, however, no relevant reports have been reported at this time. In the light of the above-mentioned problems, the structure is selected to replace 13 kinds of coumarin compounds, and the interaction between the above-mentioned compounds and the AChE is studied by using a variety of spectral methods and the molecular simulation technology. In this paper, the chemical and spatial-binding modes of the interaction between the compounds and the AChE are discussed. The effect of the substitution of the different structures on the interaction of the compounds with the AChE is compared. The enzyme activity of the AChE is studied by Elman's method. The effect of substitution of different structures on the activity of AChiE in this kind of compound was studied. In response, the results of this study are useful for finding the active groups of these compounds and to screen new type of coumarin AChiE inhibitors. I. Specific contents such as In this paper, the interaction of 13 kinds of coumarin compounds with AChE was studied by fluorescence and ultraviolet-visible absorption spectrometry, and the interaction mechanism was discussed. the combination constant, the combination of the number of bits, and the combined distance of the two The results show that the compounds have a strong quenching effect on the endogenous fluorescence of AChE. The quenching mechanism is mainly to form the static quenching of the complex, and can cause the change of the conformation of the AChE, the hydrophobic force and the van der Waals force are the main factors to interact with the AChE. the interaction distance between the compound and the AChE is less than 7 nm; the ratio of the interaction size of the different structures to the compound It is shown that the substitution of the hydroxyl groups in the 4-position of the coumarin and the substitution of the methoxy and ethoxy groups on the 7-position can be used to enhance the interaction of the compound with the AChE. A molecular simulation study on the interaction of 2-coumarines and AChE was carried out. The three-dimensional space structure of Choline esterase was used as a template protein, and the interaction of AChE with the studied coumarin-based molecules was carried out by Audiock4.0 software. The interaction energy, the binding site and the type of force are determined. The results show that the coumarin compounds can interact with the active sites of the AChiE, and the amino acids that interact with the AChiE are mainly hydrophobic and charged amino acids, which are formed in the coumarin-like molecules and the AChiE. In the complex, the van der Waals force and the hydrophobic force are dominant, and the hydrogen bond and the static force are present at the same time, and the judgment of the force type in the interaction is basically the same as that of the fluorescence method in the molecular docking study; and the coumarins in the coumarin compound The presence of a prime ring, which may be the same as that of the AChE residue, may also be the result of the AChE fluorescence The reason of the quenching is that the number of amino acid residues which can interact with the AChE is different due to the different structure, position or number of the substituents, and the number of hydrogen bonds formed by the residues is different, and the experimental results show that the substitution of the hydroxyl in the 4-position is formed. The changes of the activity of the 3-coumarin compound to the activity of AChE were studied. The activity of AChE was evaluated by Ellman, and the activity of AChE was evaluated by Ellman. The results show that the compound has a certain inhibitory effect on the activity of AChE, the substitution of the 4 and 7 positions of the coumarin shows better AChiE inhibitory activity, the substitution of the hydroxyl groups at the 7-position and the increase of the number of the hydroxyl groups make the coumarin compounds inhibit the AChE.
【學(xué)位授予單位】：鄭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2014
【分類號】：R969.2
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