| Size | Price | Stock | Qty |
|---|---|---|---|
| 2mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg |
|
||
| Other Sizes |
|
Purity: ≥98%
SCH772984 (SCH-772984) is a novel, potent and ATP-competitive inhibitor of ERK1/2 with potential antitumor activity. In a cell-free assay, it inhibits ERK1/2 with IC50 values of 4 nM and 1 nM, respectively. An affinity-based mass spectrometry high-throughput platform with potential anticancer activity found SCH772984. Out of the 300 kinases tested, SCH772984 is highly selective against them, causing more than 50% inhibition at a concentration of 1 μmol/L for kinases like CLK2, FLT4, GSG2, MAP4K4, MAPK1, MINK1, PRKD1, and TTK.
| Targets |
ERK2 (IC50 = 1 nM); ERK1 (IC50 = 4 nM)
|
|---|---|
| ln Vitro |
SCH772984 is a novel, selective and ATP competitive inhibitor of ERK1/2. The phosphorylation of p90 ribosomal S6 kinase, an ERK substrate, is inhibited by SCH772984 in a dose-dependent manner (T359/S363 phospho-RSK). Additionally, phosphorylation of ERK's own activation loop residues is inhibited by SCH772984. SCH772984 exhibits EC50 values <500 nM in approximately 88% and 49% of BRAF-mutant or RAS-mutant tumor lines, respectively. Importantly, SCH772984 successfully inhibited MAPK signaling and cell proliferation in tumor cells that were resistant to concurrent treatment with BRAF and MEK inhibitors. [1]
|
| ln Vivo |
SCH772984 induces tumor regressions in xenograft models at tolerated doses. In models with resistance to BRAF or MEK inhibitors, SCH772984 successfully inhibits MAPK signaling and cell proliferation. [1]
SCH772984 elevated blood glucose, increased the G6Pase and PEPCK expression, and inhibited pERK1/2 and pFoxo1 expression in LPS-induced mice. [2] In vivo antitumor efficacy of SCH772984 was evaluated in BRAF- or KRAS-mutant xenograft models established from human melanoma or pancreatic carcinoma cell lines. Treatment of BRAF-mutant LOX melanoma xenografts with SCH772984 (50 mg/kg twice daily) led to 98% tumor regression (Fig. 2B). Dose-dependent antitumor activity was also observed in the KRAS-mutant pancreatic MiaPaCa model, with 36% regression at 50 mg/kg twice daily (Fig. 2C). Importantly, tumor regression was accompanied by robust inhibition of ERK phosphorylation in tumor tissue (Fig. 2D). SCH772984 was well tolerated on this schedule as measured by morbidity, lethality, or body weight loss (Fig. 2E).[1] |
| Enzyme Assay |
ERK2 IMAP Enzymatic Assay[1]
SCH772984 was tested in 8-point dilution curves in duplicate against purified ERK1 or ERK2. The enzyme was added to the reaction plate and incubated with the compound before adding a solution of substrate peptide and ATP. Fourteen microliters of diluted enzyme (0.3 ng active ERK2 per reaction) was added to each well of a 384-well plate. The plates were gently shaken to mix the reagents and incubated for 45 minutes at room temperature. The reaction was stopped with 60 μL of IMAP Binding Solution (1:2,200 dilutions of IMAP beads in 1× binding buffer). The plates were incubated at room temperature for an additional 0.5 hours to allow complete binding of phosphopeptides to the IMAP beads. On the LJL Analyst, plates are read. Kinase Panel Screening[1] The kinase inhibitory profile of SCH772984 was evaluated over a 310-kinase panel at a CRO company. TdF Assay[1] The unfolding of rat ERK2, rat phosphorylated ERK2, and human MEK1 proteins was conducted at a final concentration of 0.5 μmol/L each in the TdF assay buffer (10 mmol/L HEPES pH 7.4, 150 mmol/L NaCl, 5 mmol/L MgCl2, 1 mmol/L DTT) containing a final concentration of 5× Sypro Orange reporter dye. The samples contained 10, 5, or 2.5 μmol/L compound at a final DMSO concentration of 2%. Samples were run in 5 μL in a white 384-well real-time PCR plate. The plates were sealed with a clear sealing film and assayed in the LightCycler 480-II. The temperature was ramped from 23°C to 99°C in 15 minutes. The fluorescent intensity was collected at 15 data points per degree with excitation at 465 nm and emission at 580 nm. |
| Cell Assay |
Cells are plated at a density of 4,000 per well for 96-well cell proliferation experiments (six replicates). Cells are treated with DMSO or 9 point IC50 dilution (0.001-10 μM) at 1% DMSO final for all concentrations 24 hours after cell seeding. Viability is assessed using a ViaLight luminescence kit five days after dosing, as per the manufacturer's instructions. CellTiterGlo luminescent cell viability assay is used to measure the viability of cell lines after they have been treated with SCH772984 for 4 days.[1]
Cell proliferation experiments were carried out in a 96-well format (six replicates), and cells were plated at a density of 4,000 cells per well. At 24 hours after cell seeding, cells were treated with DMSO or a 9-point IC50 dilution (0.001–10 μmol/L) at a final concentration of 1% DMSO for all concentrations. Viability was assayed 5 days after dosing using the ViaLight luminescence kit following the manufacturer's recommendations (n = 6, mean ± SE). For the cell line panel viability assay, cells were treated with SCH772984 for 4 days and assayed by the CellTiterGlo luminescent cell viability assay. For IncuCyte analysis, cells were plated as above in 96-well plates, and image-based cell confluence data were collected every 2 hours during live growth. For engineered resistant lines, cells were infected with lentivirus produced from lentiORF constructs (pLOC vector) expressing either RFP, KRASG13D, BRAFV600E, truncated BRAFV600E lacking exons 2–8 (Δ2-8), MEK1P124L, MEK1F129L, or constitutively active MEK1DD (S218D+S222D). Cells were selected in blasticidin (20 μg/mL) and used for ViaLight assays as described above. |
| Animal Protocol |
Nude mice
12.5 mg/kg, 25 mg/kg, 50 mg/kg i.p. Xenograft Tumor Growth Assay[1] Nude mice were injected subcutaneously with specific cell lines, grown to approximately 100 mm3, randomized to treatment groups (10 mice/group), and treated intraperitoneally with either SCH772984 or vehicle according to the dosing schedule indicated in the figure legends. Tumor length (L), width (W), and height (H) were measured during and after the treatment periods by a caliper twice weekly on each mouse and then used to calculate tumor volume using the formula (L × W × H)/2. Animal body weights were measured on the same days twice weekly. Data were expressed as mean ± SEM. Upon completion of the experiment, vehicle- and SCH772984-treated tumor biopsies were processed for Western blot analysis.[1] |
| References | |
| Additional Infomation |
SCH772984 is a member of the class of indazoles that is 1H-indazole substituted by pyridin-4-yl and {[(3R)-1-(2-oxo-2-{4-[4-(pyrimidin-2-yl)phenyl]piperazin-1-yl}ethyl)pyrrolidin-3-yl]carbonyl}amino groups at positions 3 and 5, respectively. It is a potent inhibitor of mitogen-activated protein kinases ERK1 and ERK2 (IC50 = 4 and 1 nM, respectively) that exhibits anti-cancer properties. It has a role as an antineoplastic agent, an EC 2.7.11.24 (mitogen-activated protein kinase) inhibitor, an apoptosis inducer and an analgesic. It is a member of pyridines, a biaryl, a secondary carboxamide, a pyrrolidinecarboxamide, a member of indazoles, a tertiary amino compound, a N-acylpiperazine, a N-arylpiperazine, a member of pyrimidines, a N-alkylpyrrolidine and a tertiary carboxamide.
The high frequency of activating RAS or BRAF mutations in cancer provides strong rationale for targeting the mitogen-activated protein kinase (MAPK) pathway. Selective BRAF and MAP-ERK kinase (MEK) inhibitors have shown clinical efficacy in patients with melanoma. However, the majority of responses are transient, and resistance is often associated with pathway reactivation of the extracellular signal-regulated kinase (ERK) signaling pathway. Here, we describe the identification and characterization of SCH772984, a novel and selective inhibitor of ERK1/2 that displays behaviors of both type I and type II kinase inhibitors. SCH772984 has nanomolar cellular potency in tumor cells with mutations in BRAF, NRAS, or KRAS and induces tumor regressions in xenograft models at tolerated doses. Importantly, SCH772984 effectively inhibited MAPK signaling and cell proliferation in BRAF or MEK inhibitor-resistant models as well as in tumor cells resistant to concurrent treatment with BRAF and MEK inhibitors. These data support the clinical development of ERK inhibitors for tumors refractory to MAPK inhibitors.[1] |
| Molecular Formula |
C33H33N9O2
|
|
|---|---|---|
| Molecular Weight |
587.67
|
|
| Exact Mass |
587.275
|
|
| Elemental Analysis |
C, 67.44; H, 5.66; N, 21.45; O, 5.44
|
|
| CAS # |
942183-80-4
|
|
| Related CAS # |
|
|
| PubChem CID |
24866313
|
|
| Appearance |
Yellow Solid powder
|
|
| Density |
1.4±0.1 g/cm3
|
|
| Boiling Point |
857.3±65.0 °C at 760 mmHg
|
|
| Flash Point |
472.3±34.3 °C
|
|
| Vapour Pressure |
0.0±3.2 mmHg at 25°C
|
|
| Index of Refraction |
1.694
|
|
| LogP |
2.1
|
|
| Hydrogen Bond Donor Count |
2
|
|
| Hydrogen Bond Acceptor Count |
8
|
|
| Rotatable Bond Count |
7
|
|
| Heavy Atom Count |
44
|
|
| Complexity |
957
|
|
| Defined Atom Stereocenter Count |
1
|
|
| SMILES |
N(C1C=CC2=C(C(C3C=CN=CC=3)=NN2)C=1)C([C@@H]1CCN(CC(N2CCN(C3C=CC(C4N=CC=CN=4)=CC=3)CC2)=O)C1)=O
|
|
| InChi Key |
HDAJDNHIBCDLQF-RUZDIDTESA-N
|
|
| InChi Code |
InChI=1S/C33H33N9O2/c43-30(42-18-16-41(17-19-42)27-5-2-24(3-6-27)32-35-11-1-12-36-32)22-40-15-10-25(21-40)33(44)37-26-4-7-29-28(20-26)31(39-38-29)23-8-13-34-14-9-23/h1-9,11-14,20,25H,10,15-19,21-22H2,(H,37,44)(H,38,39)/t25-/m1/s1
|
|
| Chemical Name |
(3R)-1-[2-oxo-2-[4-(4-pyrimidin-2-ylphenyl)piperazin-1-yl]ethyl]-N-(3-pyridin-4-yl-1H-indazol-5-yl)pyrrolidine-3-carboxamide
|
|
| Synonyms |
|
|
| HS Tariff Code |
2934.99.9001
|
|
| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
|
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.4 mg/mL (4.08 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 1.43 mg/mL (2.43 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 14.3 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. View More
Solubility in Formulation 3: 1.43 mg/mL (2.43 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Solubility in Formulation 4: ≥ 1.43 mg/mL (2.43 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 14.3 mg/mL clear DMSO stock solution to 900 μL corn oil and mix evenly. Solubility in Formulation 5: 5% DMSO+30% PEG 300+ddH2O: 0.6mg/mL Solubility in Formulation 6: 20 mg/mL (34.03 mM) in 20% SBE-β-CD adjusted to pH 4-4.5 with 1 N acetic (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.7016 mL | 8.5082 mL | 17.0164 mL | |
| 5 mM | 0.3403 mL | 1.7016 mL | 3.4033 mL | |
| 10 mM | 0.1702 mL | 0.8508 mL | 1.7016 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
Biochemical and cell-based activity of SCH772984.A, chemical structure of SCH772984.B, effects of SCH772984 on kinase activity of ERK1, ERK2, and MEK1.C, SCH772984 inhibits ERK and RSK phosphorylation.D, TdF binding activity of SCH772984, VTX-11e, or GSK1120212 to recombinant ERK2 and MEK1 enzymes.Cancer Discov.2013 Jul;3(7):742-50. |
SCH772984 is efficacious inBRAF- orRAS-mutant tumor cells.
SCH772984 is efficacious in BRAF and MEK combination resistance inBRAF-mutant A101D melanoma cells.Cancer Discov.2013 Jul;3(7):742-50. |
SCH772984 is efficacious in tumor cell lines refractory to either BRAF or MEK inhibitors.Cancer Discov.2013 Jul;3(7):742-50. |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
