Kazokaitė Justina, Niemans Raymon, Dudutienė Virginija, Becker Holger M, Leitāns Jānis, Zubrienė Asta, Baranauskienė Lina, Gondi Gabor, Zeidler Reinhard, Matulienė Jurgita, Tārs Kaspars, Yaromina Ala, Lambin Philippe, Dubois Ludwig J, Matulis Daumantas
Department of Biothermodynamics and Drug Design, Institute of Biotechnology, Vilnius University, Vilnius, Lithuania.
Department of Radiotherapy (The M-Lab Group), GROW - School for Oncology and Developmental Biology, Maastricht Comprehensive Cancer Centre, Maastricht University Medical Centre, Maastricht, The Netherlands.
Oncotarget. 2018 Jun 1;9(42):26800-26816. doi: 10.18632/oncotarget.25508.
Human carbonic anhydrase (CA) IX has emerged as a promising anticancer target and a diagnostic biomarker for solid hypoxic tumors. Novel fluorinated CA IX inhibitors exhibited up to 50 pM affinity towards the recombinant human CA IX, selectivity over other CAs, and direct binding to Zn(II) in the active site of CA IX inducing novel conformational changes as determined by X-ray crystallography. Mass spectrometric gas-analysis confirmed the CA IX-based mechanism of the inhibitors in a CRISPR/Cas9-mediated CA IX knockout in HeLa cells. Hypoxia-induced extracellular acidification was significantly reduced in HeLa, H460, MDA-MB-231, and A549 cells exposed to the compounds, with the values up to 1.29 nM. A decreased clonogenic survival was observed when hypoxic H460 3D spheroids were incubated with our lead compound. These novel compounds are therefore promising agents for CA IX-specific therapy.
人碳酸酐酶（CA）IX已成为一种有前景的抗癌靶点和实体缺氧肿瘤的诊断生物标志物。新型氟化CA IX抑制剂对重组人CA IX表现出高达50 pM的亲和力，对其他碳酸酐酶具有选择性，并通过X射线晶体学确定其直接结合到CA IX活性位点的Zn(II)上，诱导新的构象变化。质谱气体分析证实了在HeLa细胞中通过CRISPR/Cas9介导的CA IX基因敲除实验中抑制剂基于CA IX的作用机制。在暴露于这些化合物的HeLa、H460、MDA-MB-231和A549细胞中，缺氧诱导的细胞外酸化显著降低，IC50值高达1.29 nM。当缺氧的H460 三维球体与我们的先导化合物孵育时，观察到克隆形成存活率降低。因此，这些新型化合物有望成为CA IX特异性治疗的药物。
