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Tolvanen, T.

Publications and source records attributed to Tolvanen, T..

2 recordsLinked to original sources

Development of pyrazolopyrimidine based macrocyclic kinase inhibitors targeting AAK1

Since the outbreak of SARS-CoV-2 in recent years, our society has become more aware that zoonotic diseases pose a real threat. Therefore, the demand for small molecules that target host proteins, essential for viral entry and replication, has increased as an interesting strategy for the development of antiviral agents, as these agents may be effective against several different pathogens. NAK kinases is one such potential target family because they are involved in a variety of cellular functions, hijacked by viruses to invade host cells, such as clathrin-mediated endocytosis. A large number of different inhibitors have already been reported targeting NAK kinases, but there are still no compounds that selectively target AAK1 over other NAK family members, in particular the closely related family member BIKE. Here, we developed a series of pyrazolo[1,5-a]pyrimidine-based macrocyclic NAK inhibitors, starting from the acyclic AAK1 inhibitor LP-935509. Through a structure-guided activity relationship study within the NAK family, we identified potent AAK1 inhibitors 16, 18 and 27, which show promising selectivity within the NAK family. The inhibitors showed a potent inhibition of the phosphorylation of the AP-2 complex and the antiviral activity of the compounds was evaluated against various RNA viruses.

biochemistry↗

Radiosynthesis and Preclinical Evaluation of Ga-NOTA-Folate for PET Imaging of Folate Receptor β Positive Macrophages

Folate receptor {beta} (FR-{beta}) is one of the markers expressed on macrophages and a promising target for imaging of inflammation. Here, we report the radiosynthesis and preclinical evaluation of [68Ga]Ga-NOTA-folate (68Ga-FOL). First, we determined the affinity of 68Ga-FOL using human FR-{beta} expressing cells. Then, we studied atherosclerotic mice with 68Ga-FOL and 18F-FDG PET/CT. After sacrifice, the tissues excised were measured with a {gamma}-counter for ex vivo biodistribution. Further, the tracer distribution and co-localization with macrophages in aorta cryosections were studied using autoradiography, hematoxylin-eosin staining and immunostaining with anti-Mac-3 antibody. Specificity of 68Ga-FOL was assessed in a blocking study with excess of folate glucosamine. As a last step, human radiation doses were extrapolated from rat PET data. We were able to produce 68Ga-FOL at high radioactivity concentration, with high molar activity and radiochemical purity. The cell binding studies showed high (5.1 {+/-} 1.1 nM) affinity of 68Ga-FOL to FR-{beta}. The myocardial uptake of 68Ga-FOL (SUV 0.43 {+/-} 0.06) was 20-folds lower compared to 18F-FDG (SUV 10.6 {+/-} 1.8, P = 0.001). The autoradiography and immunohistochemistry of aorta revealed that 68Ga-FOL radioactivity co-localized with Mac-3-positive macrophage-rich atherosclerotic plaques. The plaque-to-healthy vessel wall ratio of 68Ga-FOL (2.44 {+/-} 0.15) was significantly higher than that of 18F-FDG (1.93 {+/-} 0.22, P = 0.005). Blocking studies verified 68Ga-FOL specificity to FR. As estimated from rat data the human effective dose was 0.0105 mSv/MBq. The organ with highest absorbed dose was kidney (0.1420 mSv/MBq). In conclusion, 68Ga-FOL is a promising new FR-{beta}-targeted tracer for imaging macrophage-associated inflammation. TABLE OF CONTENT/ABSTRACT GRAPHIC O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY

biochemistry↗