Enzyme Inhibitors as Potential Anticancer and Antiviral

A.蛋白激酶C项目合成DAG-内酯作为有效的PKC配体设计的宇宙继续扩大，以实现完全同工酶特异性的目标。今年的主要发现是：1）在固相支持物上继续合成额外的96个成员的文库，目的是最大限度地探索蛋白质-膜界面结合位点周围的化学空间。通过质谱法对文库成员的合成和表征进行了新的化学改进。2）发现了第一个靶向含有C1结构域的非激酶蛋白的特异性DAG-内酯。该化合物以低纳摩尔结合亲和力与RasGRP结合，同时显示出对PK-C同工酶的弱结合活性。这种选择性结合的生物学后果正在探索中。3)完成了具有侧链的DAG-内酯的合成，该侧链以由交替的乙炔单元和苯环构成的“刚性棒”的形式存在。初步生物学研究表明，“刚性棒”的长度与用不同脂肪酸构建的人工膜的深度之间存在相关性。如果成功，该方法将用于将DAG-内酯专门转运到质膜。4）合成的靶DAG-内酯之一在刺激α-分泌酶活性方面显示出非常有效的活性，这是治疗阿尔茨海默病的重要治疗方法。B。黄酮乙酸类似物。合成了一种具有叠氮基功能的黄酮乙酸（FAA）类似物，并显示出与母体FAA一样的活性。叠氮化物部分将用于寻找负责FAA活性的分子靶标。C. DNA甲基转移酶项目（Zebularine）。证明了zebularine 2（1H）-嘧啶酮核苷在T24膀胱癌和其他肿瘤细胞系中重新激活沉默的p16基因并使启动子区去甲基化的能力。合成了几种2 '-脱氧泽布拉林的单磷酸前药，以克服掺入DNA的低水平。前两种前药测试失败，更多的类似物正在探索中。Zebularine计划在年底前进入临床试验。

A. Protein Kinase C ProjectThe universe of synthetic DAG-lactones designed as potent PKC ligands continues to expand with the objective of achieving full isozyme specificity. The major findings this year are: 1) Syntheses of additional 96-member libraries on a solid-phase support continue with the intent to maximally explore the chemical space surrounding the binding site at the protein-membrane interface. New chemical improvements in the synthesis and characterization of the library members by mass spectrometry continue to be made.2) The first specific DAG-lactone targeting a non-kinase protein containing a C1 domain was discovered. The compound binds with low nanomolar binding affinity to RasGRP while showing weak binding activity for the PK-C isozymes. The biological consequences of this selective binding are being explored. 3) The synthesis of DAG-lactones with a side chain in the form of a "rigid rod" constructed with alternating acetylene units and benzene rings was completed Preliminary biological studies indicate a correlation between length of the "rigid rod" and depth of artificial membranes built with different fatty acids. If successful, this approach will be used to translocate DAG-lactones exclusively to the plasma membrane.4) One of the target DAG-lactones synthesized showed very potent activity in stimulating alpha-secretase activity, which is an important therapeutic approach in the treatment of Alzheimer's disease. B. Flavone acetic acid analogues. A synthetic analogue of flavone acetic acid (FAA) with an azido group designed to function as a biological reported was synthesized and shown to be as active as parent FAA. The azide moiety will be used to find the molecular target responsible for the activity of FAA. C. DNA Methyl Transferase Project (Zebularine). The ability of zebularine 2(1H)-pyrimidinone riboside to reactivate a silenced p16 gene and demethylate the promoter region in the T24 bladder carcinoma and other tumor cell lines was demonstrated. Several monophosphate pro-drug of 2'-deoxyzebularine were synthesized in order to overcome the low level of incorporation into DNA. The first two pro-drugs tested failed, and more analogues are being explored. Zebularine is scheduled to enter clinical trials before the end of the year.