A5044739463- Cancer Cells and Metastasis
- Cancer Genomics and Diagnostics
- Colorectal Cancer Treatments and Studies
- Pancreatic and Hepatic Oncology Research
- Cancer Treatment and Pharmacology
- Cancer, Hypoxia, and Metabolism
- Computational Drug Discovery Methods
- Biochemical and Molecular Research
- Cancer-related Molecular Pathways
- Protein Kinase Regulation and GTPase Signaling
- Ubiquitin and proteasome pathways
- Retinoids in leukemia and cellular processes
- Cancer therapeutics and mechanisms
- Cell Image Analysis Techniques
- Cancer Research and Treatments
- Molecular Biology Techniques and Applications
- Histone Deacetylase Inhibitors Research
- Medical Imaging Techniques and Applications
- 3D Printing in Biomedical Research
- HER2/EGFR in Cancer Research
- PI3K/AKT/mTOR signaling in cancer
- Steroid Chemistry and Biochemistry
- bioluminescence and chemiluminescence research
- Pharmacogenetics and Drug Metabolism
- Chemical Reactions and Isotopes
Moffitt Cancer Center
2003-2023
Virginia Commonwealth University
2023
University of Tampa
2013
University of South Florida
2003-2013
Abstract Currently, there are no clinically approved drugs that directly thwart mutant KRAS G12D, a major driver of human cancer. Here, we report on the discovery small molecule, KRB-456, binds G12D and inhibits growth pancreatic cancer patient-derived tumors. Protein nuclear magnetic resonance studies revealed KRB-456 GDP-bound GCP-bound conformation by forming interactions with dynamic allosteric binding pocket within switch-I/II region. Isothermal titration calorimetry demonstrated...
Both Akt 2 and acid ceramidase (ASAH1) are found aberrantly overexpressed in cancer cells, but whether these two enzymes cooperate to induce malignant transformation is not known. We that immortalized, non-transformed ectopic co-expression of Akt2 ASAH1 significantly more effective than expression each gene alone at inducing cell invasion conferring resistance apoptosis. Consistent with observations, siRNA-mediated depletion both much potent depleting inhibiting viability/proliferation...
Learning ObjectivesAfter completing this course, the reader will be able to:Describe how mutations in ras oncogenes affect cell signal transduction pathways and disrupt growth, differentiation, proliferation, survival.Identify mechanisms of action two classes prenylation inhibitors, farnesyl transferase inhibitors (FTIs) geranylgeranyl transferase-I (GGTIs).Explain vitro vivo antitumor activity FTIs GGTIs multiple myeloma myelodysplastic syndrome.Access take CME test online receive one hour...
<p>SAR studies of the inhibition KRAS G12D/GST-RBD binding. The top four confirmed hits from <a href="#fig1" target="_blank">Fig. 1</a> were all derivatives gibberellic acid and evaluated for their potency to inhibit GTP-KRAS binding by GST pulldown assays. Concentration–response experiments identified IB-21 L (KRB-456; IC<sub>50</sub> = 0.26 µmol/L) as most potent compound followed IB-21G (IC<sub>50</sub> 0.36 µmol/L), IB-21J 2.02 IIA-15D 10.82...
<p>Supplemental Figure Legends</p>
<p>KRB-456 inhibits the growth in vivo of orthotopic mt KRAS tumors derived from pancreatic cancer patients.</p>
<p>SAR studies of the inhibition KRAS G12D/GST-RBD binding. The top four confirmed hits from <a href="#fig1" target="_blank">Fig. 1</a> were all derivatives gibberellic acid and evaluated for their potency to inhibit GTP-KRAS binding by GST pulldown assays. Concentration–response experiments identified IB-21 L (KRB-456; IC<sub>50</sub> = 0.26 µmol/L) as most potent compound followed IB-21G (IC<sub>50</sub> 0.36 µmol/L), IB-21J 2.02 IIA-15D 10.82...
<p>Determination of binding affinities KRB-456 for KRAS G12D-NE, G12D NH-GTP, NH-GDP, WT-NE, G12V-NE, and G12C-NE using ITC. ITC assays were performed 2 mmol/L in the injector 100 µmol/L protein sample cell as described Materials Methods.</p>
<div>Abstract<p>Currently, there are no clinically approved drugs that directly thwart mutant KRAS G12D, a major driver of human cancer. Here, we report on the discovery small molecule, KRB-456, binds G12D and inhibits growth pancreatic cancer patient-derived tumors. Protein nuclear magnetic resonance studies revealed KRB-456 GDP-bound GCP-bound conformation by forming interactions with dynamic allosteric binding pocket within switch-I/II region. Isothermal titration calorimetry...
<p>Treatment of mice with KRB-456 inhibits P-MEK, P-AKT and P-S6 levels induces apoptosis in KRAS G12D PDXs from pancreatic cancer patients.</p>
<p>Supplemental Figure Legends</p>
<p>KRB-456 inhibits the growth of mt KRAS xenografts from patients with refractory pancreatic cancer. Fresh tumor biopsies 4 cancer were prepared as described in Materials and Methods implanted subcutaneously into NSG mice, when average volumes about 200 mm<sup>3</sup>, mice randomized treated daily vehicle (V) or KRB-456 5 mpk for 24 days, except days 12, 13 no treatment (<b>A</b>); 8, 10 19, 21 (<b>B</b>); 17 11, 12 14 to (<b>C</b>),...
<p>BI-2852 competes with KRB-456 for binding to KRAS G12D.</p>
<div>Abstract<p>Currently, there are no clinically approved drugs that directly thwart mutant KRAS G12D, a major driver of human cancer. Here, we report on the discovery small molecule, KRB-456, binds G12D and inhibits growth pancreatic cancer patient-derived tumors. Protein nuclear magnetic resonance studies revealed KRB-456 GDP-bound GCP-bound conformation by forming interactions with dynamic allosteric binding pocket within switch-I/II region. Isothermal titration calorimetry...
<p>IIA-15D shows reduced chemical shift perturbations to KRAS G12DGDP possibly due steric clash with the allosteric pocket.</p>
<p>Effects of KRB-456 on P-MEK and P-ERK levels in human pancreatic cancer Panc0203 Panc1 cells.</p>
<p>BI-2852 competes with KRB-456 for binding to KRAS G12D.</p>
<p>KRAS switch-I/II allosteric pocket is dynamic.</p>
<p>Determination of binding affinities KRB-456 for KRAS G12D-NE, G12D NH-GTP, NH-GDP, WT-NE, G12V-NE, and G12C-NE using ITC. ITC assays were performed 2 mmol/L in the injector 100 µmol/L protein sample cell as described Materials Methods.</p>
<p>Effects of KRB-456 on cell viability in human pancreatic cancer cells that harbor KRAS G12D (Panc0403 cells), G12V (Capan1 G12C (MiaPaCa2 and WT (BxPc3 cells).</p>
<p>KRB-456 inhibits the growth in vivo of orthotopic mt KRAS tumors derived from pancreatic cancer patients.</p>
<p>KRB-456 binds to KRAS G12D<sup>GDP</sup> at a switch-I/II allosteric site. <b>A,</b> Measured CSPs of <sup>15</sup>N-labeled upon KRB-456 titration up ratio 1:4 G12D<sup>GDP</sup>: are plotted as function residue number. Residues with chemical shift ranges over the significance threshold or 1.5 times colored pink and magenta, respectively. associated P-loop, switch-I switch-II structural motifs indicated yellow, blue cyan shading, Data...
<p>Supplemental Methods</p>