| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| Other Sizes |
Purity: ≥98%
GSK621 is a novel, potent and selective activator of AMP-activated protein kinase (AMPK) with potential antitumor activity. It spares healthy hematopoietic progenitors while selectively killing acute myeloid leukemia (AML) cells. Both in vitro and in vivo tests revealed strong antiproliferative activity and high antitumor efficacy.
| Targets |
AMPK
AMP-activated protein kinase (AMPK): GSK621 is a specific AMPK agonist with IC50 values of 13-30 μM against AML cell lines [1] - Autophagy: GSK621 induces autophagy through activation of AMPK in AML cells [1] - Apoptosis: GSK621 induces apoptosis in AML cells and glioma cells via caspase-dependent pathway [1][3] - eIF2α/ATF4 signaling pathway: GSK621 induces eIF2α phosphorylation (a marker of UPR activation) [1] - Tetraspanin 8 (Tspan8): In glioma cells, GSK621 downregulates Tspan8, a potential oncogene that promotes cancer progression [3] - Mammalian target of rapamycin (mTOR): GSK621 inhibits mTOR activation through AMPK in both AML and glioma cells [1][3] |
|---|---|
| ln Vitro |
GSK621 significantly increases AMPKα T172 phosphorylation, a sign of AMPK activation, in both AML cell lines and primary AML samples. GSK621 inhibits the proliferation of all 20 cell lines in the study, with IC50 values ranging from 13 to 30 μM, and increases apoptosis in 17 (85%) of the lines. Additionally, GSK621 activates autophagy, which also plays a role in AML cell death.[1]
AML cell proliferation inhibition: GSK621 (0-30 μM for 4 days) reduces proliferation of 20 different AML cell lines with IC50 values ranging from 13 to 30 μM. It increases apoptosis in 17 (85%) of these lines [1] - Autophagy induction: GSK621 (30 μM for 24 hours) induces formation of numerous cytoplasmic vacuoles including autophagosomes in AML cells and primary AML samples [1] - ER stress response: GSK621 treatment induces PERK phosphorylation (a marker of ER stress) in AML cells [1] - AMPK activation: GSK621 (30 μM) induces phosphorylation of AMPKα T172, ACC (S79) and ULK1 (S555) in AML cells [1] - Glioma cell inhibition: GSK621 inhibits survival and proliferation of glioma cells (U87MG and U251MG) in a concentration- and time-dependent manner, with IC50 values in the 10-100 μM range. It significantly decreases viable colonies and increases trypan blue-positive cells [3] - Apoptosis induction in glioma cells: GSK621 dose-dependently increases caspase-3 activity, histone DNA apoptosis ELISA OD, and Annexin V percentage in U87MG cells. Caspase inhibitors (z-DEVD-cho or zVAD-cho) attenuate GSK621-induced apoptosis and cytotoxicity, confirming caspase-dependent mechanism [3] - Selectivity: GSK621 shows minimal cytotoxicity against normal hematopoietic progenitors, human melanocytes, HCN-1a neuronal cells, and primary human astrocytes compared to cancer cells [1][3] |
| ln Vivo |
GSK621 (30 mg/kg, i.p.) reduces leukemia growth and significantly increases survival in nude mice with MOLM-14 xenografts by increasing AMPK activity and inducing apoptosis.[1]
AML xenograft model: GSK621 (30 mg/kg, intraperitoneal injection twice daily) reduces leukemia growth and significantly extends survival in MOLM-14 cells xenografted into nude mice compared to vehicle-treated animals or those treated with 10 mg/kg twice daily [1] - Glioma xenograft model: GSK621 (not fully described in available data) shows anti-tumor activity in glioma models, though specific details are limited [3] - TMZ sensitization: GSK621 (10 μM) significantly potentiates temozolomide (TMZ, 100 μM) sensitivity and lethality against glioma cells in vitro, suggesting potential synergistic effects in vivo [3] |
| Enzyme Assay |
HEPG2 cells are exposed to increasing doses of A-769662 or GSK621, and the phosphorylation of ACC S79 is measured using a commercial ELISA kit as a stand-in for the activation of AMPK.
AMPK activity assay using ELISA: HEPG2 cells are treated with increasing doses of GSK621 or control compound (A-769662). ACC (S79) phosphorylation is assessed as a surrogate marker for AMPK activation using a commercially available ELISA kit. Results are presented as a percentage of vehicle-treated cells [1][3] - Western blot analysis for kinase phosphorylation: Cells (AML or glioma) are treated with GSK621 (30 μM) for 6 hours. Protein extracts are prepared and subjected to SDS-PAGE, transferred to membranes, and probed with antibodies against phosphorylated AMPKα T172, ACC (S79), ULK1 (S555), PERK, eIF2α, and other relevant markers. β-actin is used as a loading control [1][3] |
| Cell Assay |
Twenty AML cell lines are subjected to log10 dilutions of GSK621, and CellTiter-Glo?assay is used to assess relative viability. Using the Prism software, the IC50 of GSK621 for each of these 20 cell lines is calculated and shown as log[C], where [C] is expressed in mol/l (M). Five different cell lines are represented in the results for each panel.
Cell proliferation assay: AML cell lines (MV4-11, OCI-AML3, OCI-AML2, HL-60, Kasumi, HEL, UT7, NB4, TF-1, KG1A, Nomop28, SKM-1, U937, YHP1, MOLM-14, Mo7e, K562, MOLM-13, EOL-1, SET-2) or glioma cells (U87MG, U251MG) are seeded in 96-well plates. GSK621 is added at concentrations ranging from 0 to 30 μM (AML) or 0 to 100 μM (glioma). After incubation for 4 days (AML) or 24-72 hours (glioma), cell viability is measured using MTT assay. IC50 values are calculated from dose-response curves [1][3] - Autophagy assay: AML cells or primary AML samples are treated with GSK621 (30 μM) for 24 hours. Cells are then examined for formation of cytoplasmic vacuoles (autophagosomes) by microscopy [1] - Apoptosis assay: Glioma cells are treated with GSK621 at various concentrations for 24-48 hours. Apoptosis is measured by caspase-3 activity assay, histone DNA apoptosis ELISA, or Annexin V binding assay. For mechanistic studies, cells are pre-treated with caspase inhibitors (z-DEVD-cho or zVAD-cho) for 1 hour before GSK621 addition [3] - Colony formation assay: Glioma cells are treated with GSK621 (10-100 μM) for 24 hours, then washed and replated at low density in fresh medium. After 7-10 days, colonies are fixed, stained, and counted [3] - AMPK knockdown studies: Lentiviral shRNA targeting AMPKα or empty vector control is transfected into cells. Stable cell lines are selected with puromycin. GSK621 (30 μM) is added for 24-48 hours, and cell viability, apoptosis, and signaling pathways are analyzed by MTT assay, Annexin V binding, and Western blot [3] - Dominant negative AMPK studies: Cells are transfected with plasmid encoding dominant negative AMPKα (T172A) or empty vector. Stable cell lines are selected. GSK621 (30 μM) is added for 24-48 hours, and cell viability and signaling pathways are analyzed [3] |
| Animal Protocol |
NUDE Mice bearing MOLM-14 Xenografts
30 mg/kg, twice daily i.p. AML xenograft model: MOLM-14 cells (AML cell line) are injected subcutaneously or intravenously into nude mice. When tumors reach a measurable size, mice are randomized into treatment groups. GSK621 is dissolved in appropriate vehicle (likely DMSO-based formulation) and administered via intraperitoneal injection at 30 mg/kg twice daily. Vehicle control groups receive the same volume of vehicle. Tumor growth is monitored by caliper measurements or bioluminescence imaging (if cells are luciferase-expressing). Survival is recorded daily [1] - Glioma xenograft model: U87MG or U251MG cells are implanted intracranially or subcutaneously into nude or SCID mice. Treatment with GSK621 (not fully described in available data) is initiated when tumors are established. Combination therapy with temozolomide (TMZ) may be included to evaluate synergistic effects [3] |
| References | |
| Additional Infomation |
GSK621 is a novel thienopyridone derivative that specifically activates AMPK [1][2] - GSK621 exhibits selective cytotoxicity against AML cells rather than normal hematopoietic progenitor cells due to a unique synthetic lethal interaction involving the simultaneous activation of AMPK and mTORC1. The lethality of AML cells can be eliminated by chemical or genetic means to remove the mTORC1 signaling pathway[1]
- In AML cells, the cytotoxicity of GSK621 involves the eIF2α/ATF4 signaling pathway, which is specifically induced by mTORC1 activation[1] - In glioma cells, GSK621 activates AMPK to inhibit mTOR and downregulates tetraspan protein 8 (Tspan8), a potential oncogene that promotes cancer progression and temozolomide resistance[3] - GSK621 may offer therapeutic opportunities for cancers with excessive mTORC1 activation, including AML and glioma[1][3] |
| Molecular Formula |
C26H20N3O5CL
|
|---|---|
| Molecular Weight |
489.907
|
| Exact Mass |
489.109
|
| Elemental Analysis |
2934.99.9001
|
| CAS # |
1346607-05-3
|
| Related CAS # |
1346607-05-3
|
| PubChem CID |
54577153
|
| Appearance |
Off-white to light yellow solid powder
|
| Density |
1.4±0.1 g/cm3
|
| Index of Refraction |
1.674
|
| LogP |
4.04
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
5
|
| Heavy Atom Count |
35
|
| Complexity |
778
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
COC1C=CC=C(N2C(=O)NC3=C(N(C(=C3)Cl)C3=CC=C(C4=CC=CC(OC)=C4O)C=C3)C2=O)C=1
|
| InChi Key |
KURYSXLJGKKDHT-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C26H20ClN3O5/c1-34-18-6-3-5-17(13-18)30-25(32)23-20(28-26(30)33)14-22(27)29(23)16-11-9-15(10-12-16)19-7-4-8-21(35-2)24(19)31/h3-14,31H,1-2H3,(H,28,33)
|
| Chemical Name |
6-chloro-5-[4-(2-hydroxy-3-methoxyphenyl)phenyl]-3-(3-methoxyphenyl)-1H-pyrrolo[3,2-d]pyrimidine-2,4-dione
|
| Synonyms |
GSK-621; GSK621; GSK 621
|
| 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) |
DMSO: ~97 mg/mL (~198.0 mM)
Water: <1 mg/mL Ethanol: <1 mg/mL |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 1.25 mg/mL (2.55 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 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. Solubility in Formulation 2: ≥ 1.25 mg/mL (2.55 mM) (saturation unknown) 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. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 12.5 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 1.25 mg/mL (2.55 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. Solubility in Formulation 4: 2% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5mg/mL Solubility in Formulation 5: 10 mg/mL (20.41 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.0412 mL | 10.2060 mL | 20.4119 mL | |
| 5 mM | 0.4082 mL | 2.0412 mL | 4.0824 mL | |
| 10 mM | 0.2041 mL | 1.0206 mL | 2.0412 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.
|
|
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA
