5-fluoropyrimidine heterocyclic compound with Wnt signal path inhibitory activity and application of 5-fluoropyrimidine heterocyclic compound
A technology of heterocyclic compounds and signaling pathways, applied in the field of medicine
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Embodiment 1
[0052] This example provides 5-fluoropyrimidine heterocyclic compounds A1-A96 with Wnt signaling pathway inhibitory activity and their synthesis methods.
[0053] (1) Heterocyclic compound A1, which is synthesized by the following method:
[0054]
[0055] 1) Synthesis of Intermediate A1-2:
[0056]Add compound A1-1 (6.9g, 40mmol), bis-pinacol borate (11.2g, 44mmol), KOAc (7.8g, 80mmol), Pd(dppf) in a 250mL round bottom flask 2 Cl 2 (1.63g, 2.0mmol) and tetrahydrofuran (100mL), nitrogen replacement, reflux stirring overnight, cooling to normal temperature, suction filtration with diatomaceous earth, the filtrate was spin-dried to obtain a black oily crude product (15.6g, content: 27%), directly use in the next step.
[0057] 2) Synthesis of intermediate A1-3:
[0058] The crude intermediate A1-2 (8g, 27%, 9.86mmol) was dissolved in dioxane / water (100mL / 20mL), p-bromobenzylamine (1.65g, 8.88mmol), potassium carbonate (2.72g, 19.73 mmol), Pd(dba) 2 (567mg, 0.99mmol) and...
Embodiment 2
[0408] In this example, the inhibitory ability of the 5-fluoropyrimidine heterocyclic compounds A1-A96 with Wnt signaling pathway inhibitory activity prepared in Example 1 on the Wnt signaling pathway was determined.
[0409] LWnt3A cells (CRL-2647, ATCC) were cultured in DMEM medium (Gibico) containing 10% fetal bovine serum (Hyclone). HEK293STF stable clone cells (HEK293 cells transfected with "Super-TopFlash" TCF fluorescent reporter plasmid) were cultured in complete medium (containing 4mML-glutamine, 1.5g / L sodium bicarbonate, 4.5g / L glucose, 6μg / L mL blasticidin and 10% fetal bovine serum in DMEM medium). When LWnt3A cells and HEK293STF stable clone cells were cultivated to 90% confluency, they were harvested separately and mixed at a ratio of 1:1. 100 μL / well of the mixed cell culture solution was added to a 96-well plate to make the final cell concentration 12,000 cells / well, and then cultured for another 24 hours. The compounds to be tested were serially diluted wit...
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