â… .Synthesis And Antifungal Activity Of Novel Triazole Compounds;â…¡.Synthesis And Immunological Studies Of N-Modified GM3 Antigens As Therapeutic Cancer Vaccines

During the past several decades, Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus and other fungal infection incidence had been rising sharply, with the anti-cancer drugs in clinical, immune inhibitors, widely used broad-spectrum antibiotics, cancer chemotherapy, radiation therapy, peritoneal dialysis, organ transplants generally carried out, and immune deficiency diseases, especially AIDS, rapid and widespread. Deep fungal infection had become a clinically serious diseases such as AIDS and cancer important cause of death, which had risen to the third largest infectious disease, threating to human life and health seriously. Antifungal drugs'researchs had been the focus of the global drug experts one of the areas. At present, the drugs of deep fungal infection were relatively small in clinic, which had two major problems: a narrow spectrum antimicrobial and a serious drug resistance. Therefore, an urgent clinical need for new safe, efficient, broad spectrum antibacterial, well tolerated antifungal drugs.Azoles are an important class of antifungal drugs that interfere with ergosterol biosynthesis by inhibiting a cytochrome P450 enzyme lanosterol 14αdemethylase (CYP51), an essential enzyme in this pathway. This leads to the depletion of ergosterol and accumulation of C-14 methylated sterols and hence disruption of membrane functions. Triazole antifungal drugs such as FCZ and ICZ now are the most widely used for the treatment of systemic fungal infections in clinic.Our subject mainly based on the role of azole antifungal drugs in lanosterol 14αdemethylase (CYP 51) three-dimensional structure and mode of research progress, azole antifungals QSAR study concluded, according to the computational docking experiments results, the new antifungal compounds retaining the triazole basic pharmacophore: 1,2,4-triazole ring, 2,4-difluorophenyl, and tertiary alcohol structures.With m-difluorobenzene as starting material, treated with chloroacetyl chloride to obtain theω-chloro-2, 4-difluorophenyl ethyl ketone through Friedel-Crafts reaction; then reacted with triazole in toluene to get 2,4-difluoro-2-(1H-1,2,4-triazole-1-yl) acetophenone, and then treat with sulfur Iraq Leader (ylid) iodide reagent trimethyl oxygen sulfur nucleophilic addition reaction to get a ring oxide, and then with methane sulfonic acid salt to get the key intermediate. After opening the ring with various amines to prepare 1-(1H-1,2,4-triazole-1-yl)-2-(2,4-difluorophenyl)-3-(N-substituted-amino)-2-propanols. In acetonitrile with potassium carbonate as acid binding agent and reacted with propargyl bromide to prepare 1-(1H-1,2,4-triazole-1-yl)-2-(2,4-difluorophenyl)-3-(N-substituted amino-N-propargyl amino)-2-propanols, and finally in the solvent DMSO, sodium azide and various substituted benzyl bromide reacted to get azides, 1-(1H-1,2,4-triazole-1-yl)-2-(2,4-difluorophenyl)-3-(N-substituted amino-N-propargyl amino)-2-propanols was added into the reaction directly without treatment.use copper sulfate and sodium ascorbic acid as catalyst, through the "Click Reaction" to prepare the various target compounds, this is the first attempt to re-introduction of a 1,2,3 - triazole ring in the other side of the nucleus through the "Click Reaction" in antifungal designs. Total 157 new triazole antifungal compounds were designed and synthesized in the side chain N atom was substituted with methyl, ethyl, propyl, butyl, cyclopropyl, isopropyl, allyl and so on. All of them were reported firstly. All the compounds were confirmed by 1H NMR, some of the compounds were confirmed by 13C NMR and MS.MICs of all title compounds were determined by the method recommended by the National Committee for Clinical Laboratory Standards(NCCLS) using RPMI1640 test medium. Eight fungi were used: Candida albicans SC5314,Cryptococcus neoformans ATCC32609,Candida parapsilosis,Candida tropicalis,Trichophyton rubrum,Candida kefyr,Candida albicans Y0109 and Aspergillus fumigatus. The MIC₈₀ values showed all compounds exhibited higher activity against nearly all fungi tested except Aspergillus fumigatus than FCZ; While compounds B10, C2, F19, F21 and G7 showed 128 times higher activity (with the MIC₈₀ value of 0.0039μg/mL) than that of FCZ against Candida albicans SC5314 in vitro; And compounds A12, A16, C16, C25, F4 and F13 showed 128 times higher activity (with the MIC₈₀ value of 0.0039μg/mL) than that of FCZ against Candida albicans Y0109 in vitro; compounds G7 showed 512 times higher activity (with the MIC₈₀ value of 0.00097μg/mL) than that of FCZ against Candida albicans Y0109 in vitro; compounds D5, E15 and F8 showed 512 times higher activity (with the MIC₈₀ value of 0.0039μg/mL) than that of FCZ against Candida kefyr in vitro; which is worth to further reacher. GM3, a sialylated trisaccharide antigen expressed by a number of tumors, is an attractive target in the design of therapeutic cancer vaccines. However, a serious problem associated with GM3 is that it is poorly immunogenic. To overcome this problem for the development of GM3-based cancer vaccines, four GM3 derivatives, including 5'-N-p-methylphenylacetyl, 5'-N-p-methoxyphenylacetyl, 5'-N-p-acetophenylacetyl and 5'-N-p-chlorophenylacetyl GM3, were synthesized and then coupled to a carrier protein, keyhole limpet haemocyanin (KLH). The resultant glycoconjugates were evaluated as vaccines in mouse and compared to the KLH conjugate of 5'-N-phenylacetyl GM3 (GM3NPhAc), a highly immunogenic GM3 derivative that was previously investigated as a vaccine candidate. All of the four new GM3 derivatives were proved to be more immunogenic than GM3NPhAc and elicit very strong T cell-dependent immune responses desirable for cancer immunotherapy. It was concluded that the new GM3 derivatives can form promising vaccine candidates that may be used to combine with cell glycoengineering for cancer immunotherapy.