Shp2 inhibitor compositions, methods for treating cancer and methods for identifying a subject with shp2 mutations
A technology of inhibitors and allosteric inhibitors, applied in the field of treating diseases or disorders of subjects, capable of solving problems such as reducing the efficacy of SHP2 mutants
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Embodiment 1
[0503]Activating mutations have differential effects on the biochemical efficacy of allosteric inhibitors
[0504]SHP2 (PTPN11) is a non-receptor protein tyrosine phosphatase and scaffold protein, which acts on the downstream of a variety of RTKs and integrates growth factor signals to promote RAS / MAPK activation. SHP2 is composed of three different domains: two SH2 domains at the N-terminus, followed by the PTP catalytic domain. In the absence of RTK signaling, SHP2 adopts a self-inhibiting conformation. Mutations that destabilize the self-inhibiting conformation are common in inherited RASopathies and certain cancers. Allosteric inhibitors that stabilize the self-suppressive conformation of wild-type SHP2 inhibit RAS / MAPK signaling and tumor growth driven by oncogenic mutations in the RAS / MAPK pathway in a xenograft model. This study explored the effect of allosteric inhibitors on activated mutant SHP2.
[0505]The tyrosine diphosphate motif incorporated into the signaling protein desta...
Embodiment 2
[0556]The biochemical sensitivity of SHP2 mutant predicts the cell sensitivity of allosteric inhibitor compound B
[0557]Generation of isogenic SHP2 expressing cell line
[0558]An experimental system is being established to test the activity of SHP2 mutants in an isogenic background (Figure 6 ). The Flp-In T-REx-293 cell line is fromObtained and put it in the 2mM L-glutamineAnd supplemented with 10% FBS1% penicillin / streptomycin100μg / mL ZeocinTMAnd 15μg / mL blasticidinHigh glucose DMEMTMIn a humid cell culture incubator, culture at 37°C and 5% CO2.
[0559]The wild-type or mutant SHP2 variants were synthesized and subcloned into the pcDNA5 / FRT / TO vector (ThermoFisher). Use X-tremegene 9 DNA Transfection ReagentAccording to the manufacturer’s instructions, combine the plasmid with pOG44 Flp recombinase expression plasmidCo-transfected into Flp-In T-REx-293 cells. When containing 2mM L-glutamine and supplemented with 10% FBS and 1% penicillin / streptomycin, 200μg / mL hygromycin BAnd 15μg / mL bla...
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