Barret J M, Ernould A P, Ferry G, Genton A, Boutin J A
Département de Cancérologie Expérimentale, Institut de Recherches Servier, Suresnes, France.
Biochem Pharmacol. 1993 Aug 3;46(3):439-48. doi: 10.1016/0006-2952(93)90520-7.
Tyrosine protein kinases (TPKs) play a major role in the transformation of cells. They are currently used as molecular targets for new generations of anticancer compounds. Numerous TPKs have been described from various tissues using either classical molecular biochemical techniques or cloning strategies. As a natural extension of these discoveries, a large number of "specific" inhibitors have been described in the literature. The major problem with these inhibitors is that there is no simple way to compare their specificity and/or selectivity from one report to another. We have set up a simple, straightforward technique to compare the inhibitory potency of 14 classical inhibitors towards six well-described and at least partially purified protein kinases. This technique is based on a new assay, easy to carry out and non-restrictive in terms of the type of protein substrate used. It permits direct comparisons between the results obtained from various sources. Data obtained showed that, when assessed in this integrated system, specificity and selectivity of many "classical" inhibitors are often weak, thus demonstrating that a universal technique such as ours is essential for the molecular screening of new protein kinase inhibitors. Compounds showing specificity for this panel of protein kinases will be more easily targeted to some defined types of oncogene and of transformed cells.
酪氨酸蛋白激酶（TPKs）在细胞转化过程中发挥着主要作用。它们目前被用作新一代抗癌化合物的分子靶点。使用经典分子生化技术或克隆策略，已经从各种组织中描述了众多的TPKs。作为这些发现的自然延伸，文献中描述了大量的“特异性”抑制剂。这些抑制剂的主要问题在于，没有简单的方法可以在不同报告之间比较它们的特异性和/或选择性。我们建立了一种简单、直接的技术，用于比较14种经典抑制剂对6种已充分描述且至少部分纯化的蛋白激酶的抑制效力。该技术基于一种新的检测方法，易于实施，并且在所用蛋白底物类型方面没有限制。它允许对从不同来源获得的结果进行直接比较。获得的数据表明，在这个综合系统中进行评估时，许多“经典”抑制剂的特异性和选择性往往较弱，从而表明像我们这样的通用技术对于新蛋白激酶抑制剂的分子筛选至关重要。对这组蛋白激酶具有特异性的化合物将更容易靶向某些特定类型的癌基因和转化细胞。
