| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
Grp94 (selective over Hsp90α, Hsp90β, and Trap-1; >100-fold selectivity for Grp94 over Hsp90α and Hsp90β, and 10- to 100-fold selectivity over Trap-1) [1]
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|---|---|
| ln Vitro |
PU-H54 binds to the N-terminal domain of Grp94 and induces a conformational rearrangement in the lid region, exposing a hydrophobic cleft termed Site 2. The 8-aryl group of PU-H54 inserts into Site 2, making stabilizing contacts with residues Leu104, Leu163, Phe199, Ala202, Phe203, Val211, Ile247, and Leu249. In contrast, when bound to Hsp90α, the ligand adopts a different pose with an ~80° rotation about the sulfanyl linker, and access to the equivalent cleft is blocked by Phe138. This binding mode accounts for its paralog-selective binding. [1]
PU-H54 is a representative Grp94-selective ligand from the Type 2 chemical space. [1] |
| Enzyme Assay |
Fluorescence polarization competition assays were performed to test binding to Hsp90 paralogs. Recombinant proteins (Hsp90α, Hsp90β, Grp94, Trap-1) were incubated with a fluorescent labeled Hsp90 ligand (cy3B-GM for Hsp90α, Hsp90β, and Grp94; PU-FITC3 for Trap-1) and varying concentrations of test compounds including PU-H54. The mixture was incubated at 4°C for 24 hours, and fluorescence polarization values (in mP) were measured. The inhibitor concentration at which 50% of bound fluorescent ligand was displaced (EC50) was obtained by fitting the data. [1]
Crystallization: Recombinant canine Grp94 N-terminal domain (residues 69-337 with deletion 278-327) and human Hsp90α N-terminal domain (residues 1-236) were expressed and purified. Protein-inhibitor complexes were formed by adding a twofold molar excess of PU-H54 to Grp94 or threefold molar excess to Hsp90α. Grp94-PU-H54 crystals were grown by hanging-drop vapor diffusion at 18°C using reservoir solution containing 14-17% isopropanol, 300-375 mM MgCl2, 0.1-1.0% glycerol, and 100 mM HEPES pH 7.4. Hsp90α-PU-H54 crystals were grown at 4°C using reservoir solution containing 11-15% PEG 2K MME, 200 mM MgCl2, and 100 mM sodium cacodylate pH 6.5. X-ray diffraction data were collected at SSRL beamline 11-1 using wavelength 0.979 Å. Data were reduced with HKL2000, and structures were determined by molecular replacement and refined. [1] |
| References | |
| Additional Infomation |
PU-H54 is a purine-scaffold compound. The crystal structure of Grp94-PU-H54 revealed a new allosteric binding site (Site 2) in Grp94, which is not accessible in Hsp90α or Hsp90β due to a phenylalanine side chain (Phe138 in Hsp90α, equivalent to Phe199 in Grp94) blocking the cleft. This structural insight provides a rational basis for designing paralog-selective inhibitors. [1]
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| Molecular Formula |
C18H19N5S
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|---|---|
| Molecular Weight |
337.441961526871
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| Exact Mass |
337.136
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| CAS # |
1454619-13-6
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| PubChem CID |
136236968
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| Appearance |
White to off-white solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
475.0±55.0 °C at 760 mmHg
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| Flash Point |
241.1±31.5 °C
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| Vapour Pressure |
0.0±1.2 mmHg at 25°C
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| Index of Refraction |
1.663
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| LogP |
2.3
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
24
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| Complexity |
539
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC(=C(C=C1)SC2=NC3=C(N2)C(=N)N=CN3CCCC#C)C
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| InChi Key |
GPYXPUQPGWULJH-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C18H19N5S/c1-4-5-6-9-23-11-20-16(19)15-17(23)22-18(21-15)24-14-8-7-12(2)10-13(14)3/h1,7-8,10-11,19H,5-6,9H2,2-3H3,(H,21,22)
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| Chemical Name |
8-(2,4-dimethylphenyl)sulfanyl-3-pent-4-ynyl-7H-purin-6-imine
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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 : ~100 mg/mL (~296.35 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.41 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 25.0 mg/mL clear DMSO stock solution to 900 μL of 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 | 2.9635 mL | 14.8174 mL | 29.6349 mL | |
| 5 mM | 0.5927 mL | 2.9635 mL | 5.9270 mL | |
| 10 mM | 0.2963 mL | 1.4817 mL | 2.9635 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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