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| 5mg |
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| 25mg |
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Description :GMB-475 is a novel and potent degrader of BCR-ABL1 tyrosine kinase based on PROTAC, overcoming BCR-ABL1-dependent drug resistance. GMB-475 targets BCR-ABL1 protein and recruits the E3 ligase Von Hippel Lindau (VHL), resulting in ubiquitination and subsequent degradation of the oncogenic fusion protein. GMB-475 reduced viability and increased apoptosis in primary CML CD34+ cells, with no effect on healthy CD34+ cells at identical concentrations.
| ln Vitro |
In both human CML K562 cells and murine Ba/F3 cells expressing BCR-ABL1, lead compound GMB-475 induced rapid proteasomal degradation and inhibition of downstream biomarkers, such as STAT5, and showed increased sensitivity compared with diastereomeric controls lacking degradation activity. Notably, GMB-475 inhibited the proliferation of certain clinically relevant BCR-ABL1 kinase domain point mutants and further sensitized Ba/F3 BCR-ABL1 cells to inhibition by imatinib, while demonstrating no toxicity toward Ba/F3 parental cells. Reverse phase protein array analysis suggested additional differences in levels of phosphorylated SHP2, GAB2, and SHC associated with BCR-ABL1 degradation. Importantly, GMB-475 reduced viability and increased apoptosis in primary CML CD34 cells, with no effect on healthy CD34 cells at identical concentrations. GMB-475 degraded BCR-ABL1 and reduced cell viability in primary CML stem cells.[1]
human K562 cells and murine BCR-ABL1 transformed Ba/F3 cells. GMB-475 induced the degradation of BCR-ABL1 and c-ABL1 in the context of both K562 and Ba/F3 cells with concomitant inhibition of downstream signaling via the STAT5 pathway, in a dose- and time-dependent fashion (human and murine VHL share >70% identity). In both cases, GMB-475 was capable of inhibiting cell proliferation with an IC50 of approximately 1 μM . [1] We performed dose response titrations with BCR-ABL1 transformed Ba/F3 cells for imatinib, GMB-475 and GMB-651 and found IC50 values of 0.17 μM, 1.11 μM and 1.55 μM respectively . Furthermore, we determined the IC50 of imatinib in the presence of increasing concentrations of GMB-475 or GMB-651 . Co-treatment with 2.5 μM GMB-475 reduced the IC50 of imatinib almost 3-fold, likely due to degradation reducing the BCR-ABL1 protein present, suggesting a lower dose of imatinib can entirely abrogate signaling. [2] |
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| Enzyme Assay |
Under full serum (10% FBS) conditions, both GMB-475 and GMB-651 were able to inhibit the kinase activity of BCR-ABL1, as measured by loss of pSTAT5 signal, but only GMB-475 was able to reduce phosphorylation of GAB2 and SHC . Conversely, under serum-free conditions, both GMB-475 and GMB-651 were able to inhibit phosphorylation of GAB2 and SHC, as well as STAT5 . This suggests a scaffolding role for BCR-ABL1 in signaling via this pathway. Under serum-free conditions, only the constitutively active BCR-ABL1 kinase domain is able to (auto)-phosphorylate Y177, a key docking site, and thus both degrader (GMB-475) and inhibitor (GMB-651) are able to block signaling.[2]
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| Cell Assay |
K562 cells were treated with DMSO, GMB-475 (5 μM) or GMB-651 (5 μM) for 8 h in duplicate, washed twice with PBS and lysed in RPPA lysis buffer (1% Triton X-100, 50 mM HEPES, pH 7.4, 150 mM NaCl, 1.5 mM MgCl2, 1 mM EGTA, 100 mM NaF, 10 mM Na pyrophosphate, 1 mM Na3VO4, 10% glycerol, containing freshly added protease and phosphatase inhibitors from Roche Applied Science). RPPA was performed in the MDACC CCSG core.[2]
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| References |
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| Molecular Formula |
C43H46F3N7O7S
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|---|---|
| Molecular Weight |
861.93
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| Exact Mass |
861.313
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| CAS # |
2490599-18-1
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| PubChem CID |
139600282
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
6.9
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
15
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| Rotatable Bond Count |
17
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| Heavy Atom Count |
61
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| Complexity |
1410
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| Defined Atom Stereocenter Count |
3
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| SMILES |
C1(SC=NC=1C)C1C=CC(CNC(=O)[C@@H]2CC(O)CN2C(=O)[C@H](C(C)(C)C)NC(COCCOC2C=CC(C3C=C(NC4C=CC(OC(F)(F)F)=CC=4)N=CN=3)=CC=2)=O)=CC=1
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| InChi Key |
ONDVWISBMHHLGZ-CQQKSQRMSA-N
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| InChi Code |
InChI=1S/C43H46F3N7O7S/c1-26-38(61-25-50-26)29-7-5-27(6-8-29)21-47-40(56)35-19-31(54)22-53(35)41(57)39(42(2,3)4)52-37(55)23-58-17-18-59-32-13-9-28(10-14-32)34-20-36(49-24-48-34)51-30-11-15-33(16-12-30)60-43(44,45)46/h5-16,20,24-25,31,35,39,54H,17-19,21-23H2,1-4H3,(H,47,56)(H,52,55)(H,48,49,51)/t31?,35-,39-/m0/s1
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| Chemical Name |
(2S)-1-((R)-3,3-dimethyl-2-(2-(2-(4-(6-((4-(trifluoromethoxy)phenyl)amino)pyrimidin-4-yl)phenoxy)ethoxy)acetamido)butanoyl)-4-hydroxy-N-(4-(4-methylthiazol-5-yl)benzyl)pyrrolidine-2-carboxamide
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| Synonyms |
GMB475 GMB 475 GMB-475
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| HS Tariff Code |
2934.99.9001
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| 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)
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| Solubility (In Vitro) |
DMSO : ~250 mg/mL (~290.05 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 6.25 mg/mL (7.25 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 62.5 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: ≥ 6.25 mg/mL (7.25 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 62.5 mg/mL clear DMSO stock solution to 900 μL corn oil and mix evenly. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.1602 mL | 5.8009 mL | 11.6019 mL | |
| 5 mM | 0.2320 mL | 1.1602 mL | 2.3204 mL | |
| 10 mM | 0.1160 mL | 0.5801 mL | 1.1602 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.
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