| 摘要: | 藥物設計或進一步所謂的合理藥物設計（rational drug design）是人類對抗疾病與生存環境的基本求生技能。然而，藥物設計是一複雜與困難的步驟。其中主要是因為人類對藥物分子與目標分子（蛋白）的作用行為與機制不甚了解。所以目前製藥公司基本上是以大量篩選的方式，挑出所謂先導化合物（lead compound）作用分子，再進一步修飾、改質，以設計所謂的新藥。過程中，並無太多作用機制的探討，以致造成新藥設計過程曠日費時，且可能造成相當多生理上的副作用與毒性。 本計劃我們整合了拉托維亞，立陶宛及台灣的團隊針對histone deacetylases (HDACs) 組蛋白脫乙醯酶類, matrix metalloproteinases (MMPs) 基質金屬蛋白酶and carbonic anhydrases (CAs) 碳酸酐酶等三個于細胞行為相當重要的三個蛋白酶能深入的藉由立陶宛團隊在蛋白表達、純化，拉托維亞團隊合成、改質先導化合物及台灣團隊於先導物與蛋白間之作用熱力學量測等來建立結構、能量、活性（Structural-Activity-Relationship）之基礎分子作用機制與行為，而進一步建立合理藥物設計程序（rational drug design）。 拉托維亞團隊已經合成超過30個相關之先導化合物，立陶宛團隊已有多年于微生物或動物細胞表達欲研究蛋白及相關分子結構與活性量測研究，而台灣團隊將主要以恆溫滴定微卡計(Isothermal Titration Calorimetry, ITC)量測先導化合物與蛋白之作用熱力學與建立熱力學分析模式為主，再回饋化合物合成與結構團隊，整合結構、能量、活性關係之分析模式。進一步能合理性建立藥物設計與驗證。 整個團隊于計劃相關所需的專長皆有多年經驗與傑出研發成果。研發成果也將以技轉給製藥公司為目標。 ;Design of pharmaceuticals is an essential part of human fight against various diseases. However, drug design is a multi-step and very complex process. In the first phase, the natural or synthetic compounds are discovered or identified that bind a human target protein responsible for the progression of the illness. Unfortunately, the cost of drug design has risen tremendously but the number of drugs brought to market has not grown significantly. One of the major difficulties in drug design is poor scientific understanding of protein – ligand (drug) interactions. There is no way to predict which compound would bind to which protein and with what affinity. Instead, pharmaceutical industry performs high-throughput screening of all available compounds against a particular disease target and selects the hit compounds for development into drugs. Such compounds usually bind to numerous non-intended targets thus exhibiting numerous toxicity side effects. In this proposal we combine the efforts and experience of three teams, Latvian, Lithuanian and Taiwanese in the areas of organic synthesis, recombinant protein target production and thermodynamics of protein – target interaction, respectively. The objectives of the project are: The Latvian team will design and synthesize compounds which would bind specifically and with average or high affinity to an intended target, namely, histone deacetylases (HDACs), matrix metalloproteinases (MMPs) and carbonic anhydrases (CAs). The Lithuanian team will clone, express in bacterial or human cells and purify the target proteins. Taiwanese team will experimentally determine the energies of compound binding to the three target protein groups by isothermal titration calorimetry to measure the enthalpies, entropies and the Gibbs free energies. They will also develop novel protocols and approaches to prepare compounds with better binding properties and develop the theory of thermodynamic data application for the structure-thermodynamics correlation. In addition, Lithuanian team will determine the Gibbs free energies of compound binding by the inhibition of enzymatic activity (IC50) and the fluorescent thermal shift assay for the selected best lead compounds in order to help rank the compounds for their SAR analysis. The project is possible only by combining the efforts of all three teams and the achievement of the objectives would significantly advance both the fundamental science in the area of energetics of molecular interactions and the application of inhibitors for further development into drugs. As an outcome of the project would be several lead compounds to be outlicensed to pharmaceutical companies for the development into drugs against cancer, inflammation, glaucoma and several other illnesses. |
