Kartogenin_LCMS_13149_MedChemExpress

Inhibitors, Agonists, Screening Data SheetBIOLOGICAL ACTIVITY: EPZ015666 is an orally available inhibitor of PRMT5 enzymatic activity in biochemical assays with IC50 of 22 nM and broad selectivityagainst a panel of other histone methyltransferases.IC50 & Target: IC50: 22 nM (PRMT5)[1]In Vitro: Treatment of MCL cell lines with EPZ015666 leads to inhibition of SmD3 methylation and cell death, with IC50 values in thenanomolar range[1]. EPZ015666, a potent peptide–competitive and SAM–cooperative inhibitor with >10,000–fold specificity againstPRMT5 relative to other methyltransferases[2].In Vivo: EPZ015666 is orally bioavailable and amenable to in vivo studies. We performed 21–d efficacy studies in severe combinedimmunodeficiency (SCID) mice bearing subcutaneous Z–138 and Maver–1 xenografts, with twice–daily (BID) oral dosing on fourdose groups: 25, 50, 100 and 200 mg per kilogram of body weight (mg/kg). After 21 d of continuous dosing, animals areeuthanized, and blood and tissues are analyzed to determine the relationship between methyl–mark pharmacodynamics andtumor–growth inhibition (TGI). EPZ015666 showed dose–dependent exposure and TGI after 21 d in both MCL models. Tumors in allEPZ015666 dose groups measured on day 21 showed statistically significant differences in weight, volume and tumor growthcompared to vehicle–treated tumors. Dosing at 200 mg/kg BID induced tumor stasis in Z–138 cells, with >93% TGI after 21 d,whereas Maver–1 cells showed >70% TGI. Additionally, a third MCL xenograft is tested using the Granta–519 cell line, afast–growing model that reached endpoint on day 18 and showed dose–dependent efficacy with 45% TGI in the 200 mg/kg group.EPZ015666 is well tolerated in all three models, with minimal bodyweight loss in the 200 mg/kg dose group and no other clinicalobservations[1].PROTOCOL (Extracted from published papers and Only for reference) Kinase Assay:[1]EPZ015666 is serially diluted threefold from 1,000 to 0.051 nM and spotted into a 384–well polypropyleneV–bottom microplate. 3H–SAM is serially diluted twofold in assay buffer for a seven–point dilution series with a top concentrationof 700 nM (final assay concentration). Reactions are initiated by the addition of 4 nM enzyme and 40 nM peptide (final assayconcentrations for both). Reactions are incubated for 60 min and quenched by the addition of 10 μL per well of 600 μM unlabeledSAM in assay buffer (final assay concentration). For the peptide competition, EPZ015666 is serially diluted threefold from 1,000 to0.051 nM and spotted into a 384–well polypropylene V–bottom microplate. Peptide is serially diluted twofold in assay buffer for aseven–point dilution series with a top concentration of 480 nM (final assay concentration). Reactions are initiated by the addition of4 nM enzyme and 75 nM 3H–SAM (final assay concentrations for both). Reactions are incubated for 60 min, and the reactions arequenched by the addition of 10 μL per well of 600 μM unlabeled SAM in assay buffer (final assay concentration)[1].Cell Assay: EPZ015666 is dissolved in DMSO and stored, and then diluted with appropriate medium (final DMSO 0.2%)before use[1].[1]Cultured cells in linear/log–phase growth are split to a seeding density of 2×105 cells/mL in 2–20 mL of media,depending on the yield required at the end of the growth period. Compound is diluted in DMSO and added to each culture vesselProduct Name:EPZ015666Cat. No.:HY-12727CAS No.:1616391-65-1Molecular Formula:C20H25N5O3Molecular Weight:383.44Target:Histone MethyltransferasePathway:EpigeneticsSolubility:DMSO

Product Name:InolitazoneCAS No.: 223132-37-4CatNo:HY-14792Product Data Sheet

Cat. No.:HY14792MWt:502.58Formula:C27H26N4O4SPurity :>98%

Solubility:

Mechanisms:

Biological Activity: Pathways:Cell Cycle/DNA Damage; Target:PPAR Pathways:NF-KB; Target:PPAR

DMSO

Description: IC50 Value: 0.8 nM [1]Inolitazone(RS5444) is a novel high-affinity PPARgamma agonist, which activates PPARgammawith an EC50 about 1/50th that of rosiglitazone and has no effect on RIE cells that do not express PPARgamma.in vitro: In vitro, the IC50 for growth inhibition is approximately 0.8 nM while anaplastic thyroidcarcinoma (ATC) tumor growth was inhibited three- to fourfold in nude mice. siRNA againstPPARgamma and a pharmacological antagonist demonstrated that functional PPARgamma wasrequiredforgrowthinhibitoryactivityofRS5444RS5444upregulatedthecellcyclekinaseinhibitor

Inhibitors, Agonists, Screening Certificate of Analysis

PHYSICAL AND CHEMICAL PROPERTIES Molecular Formula:C26H29NOMolecular Weight:371.51Storage:Powder-20°C3 years4°C2 yearsIn solvent-80°C6 months-20°C1 monthChemical Structure:

ANALYTICAL DATA Appearance:White to off-white (Solid)1 H NMR Spectrum:Consistent with structureLCMS:Consistent with structurePurity (LCMS):99.23%Conclusion:The product has been tested and complies with the given specifications.Product Name:TamoxifenCat. No.:HY-13757ACAS No.:10540-29-1Batch No.:23623Chemical Name:Ethanamine, 2-[4-[(1Z)-1,2-diphenyl-1-buten-1-yl]phenoxy]-N,N-dimethyl-

Caution: Product has not been fully validated for medical applications. For research use only.Tel: 609-228-6898 Fax: 609-228-5909 E-mail: tech@Address: 1 Deer Park Dr, Suite Q, Monmouth Junction, NJ 08852,

Inhibitors, Agonists, Screening Data Sheet

BIOLOGICAL ACTIVITY: Sotrastaurin is a potent pan–PKC inhibitor, with Kis of 0.22 nM, 0.64nM, 0.95 nM, 1.8 nM, 2.1 nM and 3.2 nM for PKCθ, PKCβ, PKCα,PKCη, PKCδ and PKCε, respectively.IC50 & Target: IC50: 0.22 nM (PKCθ), 0.64 nM (PKCβ), 0.95 nM (PKCα), 1.8 nM (PKCη), 2.1 nM (PKCδ), 3.2 nM (PKCε)[1]In Vitro: In cell–free kinase assays Sotrastaurin (AEB071) inhibits PKC, with Ki values in the subnanomolar to low nanomolarrange. When Sotrastaurin is tested on a selected panel of kinases, the only enzyme on which Sotrastaurin displays an IC50valuebelow 1 μM is glycogen synthase kinase 3β[1]. Sotrastaurin (AEB071) inhibits p–MARCKS, a PKC substrate, and pS6 in all the celllines, independently of the mutational status. There is a slight inhibition of pERK at lower doses also in the GNA11 mutant cells, butnot in the WT cells at any concentrations. This is consistent with previous reports indicating that Sotrastaurin inhibits ERK1/2phosphorylation in GNAQ mutant cell lines[2].In Vivo: The combination therapy results in a significantly enhanced reduction in tumor volume when compared to eitherSotrastaurin (AEB071) or BYL719 alone (p=0.049 vs. BYL719 and p=0.022 vs. Sotrastaurin at day 26). There is even a greater effectwhen compared to vehicle control (p=0.016)[2]. Sotrastaurin (STN) treatment of liver donors and orthotopic liver transplantation(OLT) recipients (Gr.I) or of OLT recipients alone (Gr.II) prolongs animal survival, as 9 out of 10 rats in Gr. I, and 6 out of 6 rats in Gr.IIsurvive >14 days. In contrast, only 4 out of 10 control OLT recipients remain alive at day 14 (p<0.01)[3]. PROTOCOL (Extracted from published papers and Only for reference) Kinase Assay:[1]Classical and novel PKC isotypes are assayed by scintillation proximity assay technology. In brief, the assay isperformed in 20 mM Tris–HCl buffer, pH 7.4, and 0.1% bovine serum albumin by incubating 1.5 μM of the peptide substratewith 10 μM [33P]ATP, 10 mM Mg (NO3)2, 0.2 mM CaCl2, and PKC at a protein concentration varying from 25 to 400 ng/mL, and lipidvesicles containing 30 mol% phosphatidylserine, 5 mol% diacylglycerol (DAG), and 65 mol% phosphatidylcholine at a final lipidconcentration of 0.5 μM. Incubation is performed for 60 min at room temperature. The reaction is stopped by adding 50 μL of amixture containing 100 mM EDTA, 200 μM ATP, 0.1% Triton X–100, and 0.375 μg/well streptavidin–coated scintillation proximityassay beads in PBS without Ca2+ and Mg2+. Incorporated radioactivity is measured in a MicroBetaTrilux counter for 1 min. PKCζ isassayed. In situ Thr–219 autophosphorylation status analysis of PKCθ is done by a phospho–site–specific antibody[1].Cell Assay: Sotrastaurin (AEB071) is dissolved in DMSO (10 mM) and stored (-20°C), and then diluted with appropriate mediabefore use[2].[2]Cells are plated in a 96–well plate and treated with Sotrastaurin, BYL719 or DMSO at indicated concentrations for aperiod of 5 days. Viability is assessed using Cell Counting Kit. The Combination Index values are calculated using the CompuSynsoftware. Briefly explained, the plots generated by the CompuSyn software demonstrate the Y–axis combination index values,where CI<1, =1, and >1 indicate synergism, additive effect, and antagonism, respectively. The X–Axis represents the fractionalactivity, which reflects the fraction of cells inhibited by the treatments relative to vehicle control. For combination index studies, theconcentrations tested included Sotrastaurin (0, 125, 250, 500, 1000 nM) and BYL719 (0, 250, 500, 1000, 2000 nM)[2].Product Name:SotrastaurinCat. No.:HY-10343CAS No.:425637-18-9Molecular Formula:C25H22N6O2Molecular Weight:438.48Target:PKC; PKCPathway:TGF–beta/Smad; EpigeneticsSolubility:10 mM in DMSO