| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg |
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| 50mg | |||
| Other Sizes |
| Targets |
CK1α (Kd = 210 nM) [1]
CK1δ (Kd = 340 nM) [1] CK1ε (Kd = 300 nM) [1] |
|---|---|
| ln Vitro |
In DiscoveRx KinomeScan interaction assays, (S)-CR8 showed binding to CK1α (score 2.6, Kd 210 nM), CK1δ (score 3.2, Kd 340 nM), and CK1ε (score 0.35, Kd 300 nM), as well as to CK1γ1, CK1γ2, and CK1γ3 with lower affinity (Kd 1300 nM each). [1]
In vitro, (S)-CR8 is reported to be ~100-fold more potent than roscovitine at inducing tumor cell apoptosis (reference 5,6 within the paper). [1] (S)-CR8 was more potent than roscovitine at reducing cystogenesis in an orthologous model of ADPKD (reference 13 within the paper). [1] |
| ln Vivo |
In an orthologous ADPKD mouse model (Pkd1 conditional knockout or other models), (S)-CR8 effectively blocked renal and hepatic cystogenesis (reference 13 within the paper). The effect was superior to that of roscovitine. [1]
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| Enzyme Assay |
Kinase interaction panel (DiscoveRx KinomeScan): For most assays, kinase-tagged T7 phage strains were grown in E. coli, lysed, and filtered. Streptavidin-coated magnetic beads were treated with biotinylated small molecule ligands (including (S)-CR8) to generate affinity resins. Binding reactions were assembled with kinases, liganded beads, and test compounds in binding buffer (20% SeaBlock, 0.17× PBS, 0.05% Tween 20, 6 mM DTT). An 11-point threefold serial dilution of each compound (top concentration 30 μM) was used. After 1 h incubation at room temperature, beads were washed four times with wash buffer (1× PBS, 0.05% Tween 20), then eluted with buffer containing non-biotinylated ligand. Kinase concentration in eluates was measured by quantitative PCR. Kd values were determined from 11-point dose-response curves performed in duplicate. [1]
Affinity chromatography on immobilized (S)-CR8: CNBr-activated sepharose beads were coupled with (S)-CR8 plus a linker at 20 mM. Kidney extracts (700 μg total protein) were loaded onto beads, incubated at 4°C for 30 min, washed four times with bead buffer (50 mM Tris pH 7.4, 250 mM NaCl, 5 mM EDTA, 5 mM EGTA, 5 mM NaF, 0.1% Nonidet P-40, protease inhibitors). Bound proteins were eluted with SDS-PAGE loading buffer and analyzed by Western blot. [1] CK1 kinase activity assay: CK1ε or CK1α were immunoprecipitated from kidney lysates using specific antibodies and protein G sepharose. Immunoprecipitates were washed with buffer C (25 mM MOPS pH 7.2, 5 mM EGTA, 15 mM MgCl2, 60 mM β-glycerophosphate, 30 mM p-nitrophenylphosphate, 2 mM DTT, 0.1 mM sodium orthovanadate, 1 mM phenylphosphate disodium). Kinase activity was assayed in buffer C with 50 μM CK-S peptide (RRKHAAlGpSAYSITA) and 15 μM cold ATP plus [γ-33P]ATP (3000 Ci/mmol) in 30 μl total volume for 30 min at 30°C. Reaction was spotted onto P81 phosphocellulose filters, washed with 1% phosphoric acid, and radioactivity counted by scintillation. [1] |
| Cell Assay |
For affinity chromatography, kidney extracts from healthy and polycystic (human ADPKD or jck mouse) were prepared and loaded onto (S)-CR8-agarose beads. After washing, bound proteins were resolved by SDS-PAGE and Western blotted with antibodies against CK1ε or CK1α. This demonstrated increased CK1ε binding and altered CK1α isoform pattern in PKD vs. healthy kidneys. [1]
For CK1 immunoprecipitation, kidney extracts (400 μg protein) were precleared with protein G sepharose, then incubated with anti-CK1ε or anti-CK1α antibodies (2 μg) followed by protein G beads. Immunoprecipitates were analyzed by Western blot. [1] |
| Animal Protocol |
The animal models used in this study included multiple PKD mouse strains (jck, bpk, Pkd1-cKO, Pkd2-cKO, etc.) and the non-cystic Nphp4-KO model. However, no specific in vivo dosing protocol for (S)-CR8 is described in this paper. The paper cites previous work (Bukanov et al., 2012) for the in vivo efficacy of CR8. That reference described oral administration of CR8 in an ADPKD mouse model. [1]
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| Toxicity/Toxicokinetics |
Systemic administration of (S)-CR8 (5 mg/kg i.p.) did not affect any physiologic parameters (oxygen saturation, breathing rate, heart rate) in rats after LFP injury. No other toxicity data reported. [2]
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| References |
[4].Casein kinase 1\u03b5 and 1\u03b1 as novel players in polycystic kidney disease and mechanistic targets for (R)-roscovitine and (S)-CR8. Am J Physiol Renal Physiol. 2018 Mar 14. |
| Additional Infomation |
(2S)-2-[[9-propyl-2-yl-6-[[4-(2-pyridyl)phenyl]methylamino]-2-purine]amino]-1-Butidrine is a phenylpyridine.
(S)-CR8 is a more potent analog of roscovitine, initially developed as a CDK inhibitor. It targets CK1α, CK1δ, and CK1ε in addition to CDKs. In polycystic kidney disease, CK1ε is overexpressed at mRNA and protein levels, and CK1α shows an altered isoform pattern with increased catalytic activity. The beneficial effects of (S)-CR8 on cystogenesis are proposed to result from dual inhibition of CDKs (CDK2, CDK5, CDK7, CDK9) and CK1s, leading to reduced proliferation, increased apoptosis, and modulation of Wnt, Hedgehog, and mTOR signaling pathways. [1] |
| Exact Mass |
431.243
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|---|---|
| CAS # |
1084893-56-0
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| Related CAS # |
(R)-CR8;294646-77-8
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| PubChem CID |
25211051
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
3.759
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
32
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| Complexity |
557
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CC[C@@H](CO)NC1=NC(=C2C(=N1)N(C=N2)C(C)C)NCC3=CC=C(C=C3)C4=CC=CC=N4
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| InChi Key |
ULEOUNVVBPZETL-SFHVURJKSA-N
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| InChi Code |
InChI=1S/C24H29N7O/c1-5-18(14-32)27-24-28-22(21-23(29-24)31(15-26-21)16(2)3)30(4)19-11-9-17(10-12-19)20-8-6-7-13-25-20/h6-13,15-16,18,32H,5,14H2,1-4H3,(H,27,28,29)/t18-/m0/s1
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| Chemical Name |
(S)-2-((9-isopropyl-6-(methyl(4-(pyridin-2-yl)phenyl)amino)-9H-purin-2-yl)amino)butan-1-ol
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| Synonyms |
(S)-CR8 (S) CR8 (S)CR8
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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) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
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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Mycobacterium Ulcerans Infection | University Medical Center Groningen | 2012-12 | Phase 2 Phase 3 |
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