| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| 500mg | |||
| Other Sizes |
| Targets |
Trypanosoma (IC50 = 10 nM)
Cruzain (Cysteine protease from Trypanosoma cruzi; IC₅₀ = 0.01 μM) [1] |
|---|---|
| ln Vitro |
NMR-based titration experiments with selectively ¹⁵N-Cys, ¹⁵N-His, and ¹³C-Met labeled cruzain showed that Cruzain-IN-1 (compound 2) binds covalently and reversibly to the catalytic Cys25 residue. The chemical shift perturbation profiles were similar to those of the irreversible vinyl sulfone inhibitor K777, confirming active site binding. Significant perturbations were observed for residues at or near the active site (e.g., Cys25, His162, Met68, Met145). Precipitate formation occurred in the NMR tube during titration, suggesting the compound itself may aggregate in solution. [1]
The binding mode was verified by comparing the nitrile derivative (compound 2) with its chlorinated analog (compound 3). The nitrile group is necessary for covalent binding to Cys25, whereas the chloro derivative showed only weak, transient non-covalent interaction. [1] |
| Enzyme Assay |
NMR-based inhibitor binding assays were performed. Samples contained 0.025–0.050 mM selectively ¹⁵N-His, ¹⁵N-Cys, or ¹³C-Met labeled cruzain in NMR buffer. Inhibitors were dissolved in deuterated DMSO to make 10 mM stock solutions. Titrations were performed by adding aliquots of the stock solution stepwise until a 10–20 fold stoichiometric excess relative to cruzain was achieved. Five titration points (apo, 2.5x, 5x, 10x, and 20x molar excess) were acquired for each inhibitor. ¹⁵N-¹H HSQC and ¹³C-¹H HSQC spectra were recorded to monitor chemical shift perturbations of cysteine, histidine, and methionine residues. Total chemical shift perturbation values were calculated. [1]
|
| References | |
| Additional Infomation |
6-(3,5-difluoroaniline)-9-ethyl-2-purine nitrile belongs to the 6-aminopurine class of compounds. Cruzain-IN-1 (compound 2) is a nitrile-based covalently reversible inhibitor of the Cruzain protease. [1] It has the same chemical skeleton as compound 3 (a chlorinated intermediate), but the nitrile functional group is crucial for the covalent interaction with the catalytic cysteine (Cys25) thiol. [1] This study demonstrates that selectively isotopically labeled nuclear magnetic resonance spectroscopy can be used to rapidly verify and characterize the binding modes of inhibitors, distinguishing between covalent, non-covalent, and non-interacting compounds. [1]
|
| Molecular Formula |
C14H10F2N6
|
|---|---|
| Molecular Weight |
300.266208171844
|
| Exact Mass |
300.094
|
| CAS # |
1199523-24-4
|
| Related CAS # |
1199523-24-4
|
| PubChem CID |
44143088
|
| Appearance |
White to off-white solid
|
| LogP |
2.812
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
7
|
| Rotatable Bond Count |
3
|
| Heavy Atom Count |
22
|
| Complexity |
430
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
FC1C=C(C=C(C=1)NC1=C2C(=NC(C#N)=N1)N(C=N2)CC)F
|
| InChi Key |
SZYYBVWPURUFRR-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C14H10F2N6/c1-2-22-7-18-12-13(20-11(6-17)21-14(12)22)19-10-4-8(15)3-9(16)5-10/h3-5,7H,2H2,1H3,(H,19,20,21)
|
| Chemical Name |
6-(3,5-difluoroanilino)-9-ethylpurine-2-carbonitrile
|
| Synonyms |
Cruzain-IN-1
|
| 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: 60~80 mg/mL (199.8~266.4 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 2.67 mg/mL (8.89 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 26.7 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: ≥ 2.67 mg/mL (8.89 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 26.7 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 | 3.3303 mL | 16.6517 mL | 33.3034 mL | |
| 5 mM | 0.6661 mL | 3.3303 mL | 6.6607 mL | |
| 10 mM | 0.3330 mL | 1.6652 mL | 3.3303 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
