Synthesis And Activity Of New Statine & Carbopy-ronine Cells Of Ca <sup> 2 + </ Sup> Fluorescent Probe Design And Synthesis Of Research

The dissertation is divided into two parts.The first is synthesis and biological activity of novel statine derivatives.The second is design and study on the synthesis of carbopyronine-based cytoplasmic calcium fluorescent probes.Many aspartic proteases have important relationships with human deseases,such as AIDS,AD,ect.They play a key role in the pathogeny of the deseases.Pepstatin,which contains the novel amino acid statine,is a natural pentapeptide isolated from natrue product.It is a good inhibitor of most aspartyl proteases with a K_i in the nanomolar range.One of the major problems,however,remains the poor solubility and membrane permeation of pepstatin, as well as its lack of selectivity among aspartyl proteases.The amino acid statine can mimic the tetrahedral transition-state of amide bond hydrolysis but cannot itself be hydrolyzed by the enzyme.It binds the enzyme through noncovalent interaction(i.e.hydrogen bonds).Thus, introduction of statine has become an efficient strategy for the the design of aspartic protease inhibitors.The unique properties of the ferrocenyl group has made ferrocene and its derivatives very popular molecules for biological applications.Among them there are compounds with antiproliferative effect,antibiotics,pesticidal activity,ect.Ferrocene is ideal for use in drug design because of the low toxicity of the molecule containing a ferrocenyl moiety and ferrocene-containing compounds often possess unexpected biological activity.In view of these facts,conjugates of ferrocene with statine were designed and sythesized in solution using the standard DCC/HOBt protocol and characterized by IR,¹H NMR spectroscopy, MS and elemental analysises.Some IR,¹H NMR and MALDI-TOF MS spectroscopies have been analyzed.Several reaction conditions have been optimized and some reaction mechanisms,such as the coupling reaction using DCC,have been proposed.All the target compounds were screened against Aspergillus oryzae,ect.The results of bioassay showed that some target compounds have good activity against Aspergillus oryzae,and very low activity against Aspergillus niger,no activity againt Bacillus licheniformis and Bacillus mycoides at a concentration of 5×10^-4 M.The results laid solid theoretical foundation for further research work. The second is design and study on the synthesis of carbopyronine-based cytoplasmic calcium fluorescent probe.Calcium is one of the most abundant elements in the human body. It plays a key role in regulating numerous physiological phenomena and serves as an intracellular second messenger and mediator.Also,it has important relationships with human deseases.The measurement of cytosolic Ca²⁺ has become an important area of investigation in chemical,biological and medical research.Nowadays,the method of the measurement of cytosolic Ca²⁺ is according to Tsien et al.There are several cytosolic Ca²⁺ indicators,such as Quin-2,Indo-1,Fura-2 and Fluo-3.They have made contributions.However,they have some drawbacks.Noteably,the optical properties of carbopyronine are very similar to those of the rhodamines.However,its absorption and fluorescence are shifted remarkably towards longer wavelengths.In the thesis a novel cytosolic Ca²⁺ fluorescent probe with low affinity to calcium,long and excitation wavelength is designed.During the research,ten novel carbopyronine fluorescent dye intermediates have been synthesized and are characterized by IR,¹H NMR spectroscopy,ect.Two crystal structures are obtained.A new method of sythesis of 2,7-Bis(dimethylamino)-9,10-dihydro-9,9-dimehtylanthracene has been presented.It is an efficient and practical method for the synthesis of the compound in the presence of non-protic Lewis acid.Mild conditions in a satisfied yield were achieved.The first method for the construction of anthracene cyclic was failed and the second method is currently underway. Although the target compound is not obtained,the results of the experiments lay solid theoretical foundation for further research work.