Periodic Reporting for period 1 - MinBioTag (A minimal Cys-cyclobutene non-canonical amino acid for bioorthogonal imaging of proteins in live cells)
Reporting period: 2016-04-01 to 2018-03-31
At this point, a change in my approach was needed, so I envised a small-molecule cyclobutene conjugate to interrogate key biological aspects of cancer. As a small molecule I selected azetolamide, a ligand of carbonic anhydrase IX. Carbonic anhydrase IX is a membrane protein overexpressed in several cancers, including lung, stomach, pancreas or brain. Besides the ubiquitous presence of this protein in cancer, there is still some controversy about whether this molecule is internalised or not upon ligand binding. In order to answer this question, an azetolamide-cyclobutene conjugate was designed and synthesized. As an IEDDA partner for this conjugate a tetrazine-bodipy was synthesized. The bodipy selected had been previously used for super-resolution microscopy and we expect that the reaction between the azelotamide-cyclobutene conjugate and the tetrazine-bodipy in cells will help us to deepen our knowledge of carbonic anhydrase IX.
We can expect two main types of results from this project. First, the development of a new dienophile for IEDDA reactions that may find wide application as a tool for both chemists and biologist to interrogate biology. Second, a deeper insight into the biological processes carbonic anhydrase IX is involved. This will, hopefully, have wide impact in the development of antibody-drug conjugates directed against this target, overexpressed in tumor hypoxia and certain types of cancer as lung, pancreas or breast.