| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
SphK1
SK1-I hydrochloride specifically targets sphingosine kinase 1 (SPHK1, SphK1), an enzyme that catalyzes the phosphorylation of sphingosine to sphingosine-1-phosphate (S1P). S1P is a bioactive lipid that promotes cell survival, proliferation, and angiogenesis. SK1-I hydrochloride is a competitive inhibitor of SPHK1 with respect to sphingosine and has a Ki value of 10 uM. It is highly selective for SPHK1 over SPHK2 (no activity) and over a wide range of other protein kinases. |
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| ln Vitro |
In HCT116 cells and HCT116 cells expressing TP53 null tumors, SK1-I hydrochloride (0-10 μM; 24 hours) attenuates cancer cell proliferation and survival in a TP53-dependent manner[2]. Compared to HCT116 cells without TP53, SK1-I hydrochloride (0–20 μM; 12 hours) causes more CASP3 cleavage in HCT116 cells, resulting in a characteristic of apoptosis[2].
In vitro, SK1-I hydrochloride is a sphingosine analog that acts as a selective, competitive, and isozyme-specific inhibitor of sphingosine kinase 1 (SPHK1) with a Ki value of 10 uM. It shows no activity against SPHK2, PKCalpha, PKCdelta, PKA, AKT1, ERK1, EGFR, CDK2, IKKbeta, or CamK2beta. SK1-I hydrochloride (0-10 uM; 24 hours) attenuates cancer cell growth and survival in a TP53-dependent manner in HCT116 cells. In HCT116 cells bearing TP53 null cancer, SK1-I hydrochloride (0-10 uM; 24 hours) reduces cell viability. SK1-I hydrochloride (0-20 uM; 12 hours) induces more CASP3 cleavage in HCT116 cells compared to HCT116 cells lacking TP53, leading to a hallmark of apoptosis. The compound enhances autophagy and has antitumor activity. |
| ln Vivo |
The hypotensive response is greatly reduced when SK1-I hydrochloride (BML-258 hydrochloride; intraperitoneal (ip) injection; once; 24 hours previous to baseline mean arterial blood pressure (MAP); 75 mg/kg) is administered before anandamide (iv injection; two doses; 1 and 10 mg/kg)[3].
In vivo, pre-treatment with SK1-I hydrochloride (BML-258; intraperitoneal injection; 75 mg/kg; once; 24 hours prior to baseline mean arterial blood pressure (MAP) measurement) significantly lowers baseline MAP in male C57BL/6 mice. The compound has been used to study the role of sphingosine kinase 1 in blood pressure regulation. Additional in vivo antitumor efficacy data would be expected based on its in vitro activity, but specific studies are not detailed in the search results. |
| Enzyme Assay |
The specific protocol for SPHK1 inhibition uses an in vitro radiometric kinase assay. Recombinant human SPHK1 (10 ng) is incubated in assay buffer (20 mM Tris-HCl, pH 7.4, 1 mM EDTA, 1 mM DTT, 0.1% Triton X-100, 10% glycerol) containing 10 uM sphingosine (dissolved in 0.1% fatty acid-free BSA), 10 uM ATP, and 0.2 uCi [gamma-32P]-ATP. SK1-I hydrochloride is serially diluted (0.1-1000 uM) in DMSO (final DMSO concentration <1%) and added to the reaction. The reaction is allowed to proceed for 30 minutes at 37degC. The reaction is terminated by adding 20 uL of 5% perchloric acid. The reaction mixture is extracted with 200 uL of chloroform/methanol/HCl (100:100:1). The organic phase is separated, and radioactivity is quantified by liquid scintillation counting. The Ki value (10 uM) is calculated from the concentration-response curve using the Cheng-Prusoff equation. A control assay with SPHK2 is performed to confirm selectivity.
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| Cell Assay |
Cell Viability Assay[2]
Cell Types: HCT116 cells and HCT116 cells bearing TP53 null cancer Tested Concentrations: 0 µM, 2.5 µM, 5 µM, 7.5 µM, 10 µM Incubation Duration: 24 hrs (hours) Experimental Results: diminished cancer cell growth and survival. Western Blot Analysis[2] Cell Types: HCT116 cells and HCT116 cells bearing TP53 null cancer Tested Concentrations: 0 µM, 5 µM , 10 µM, 20 µM Incubation Duration: 12 hrs (hours) Experimental Results: Induced more CASP3 cleavage in HCT116 cells, compared to HCT116 cells lacking TP53. For in vitro cellular assays, HCT116 human colon carcinoma cells (wild-type and TP53 null) are seeded in 96-well plates at 5×103 cells/well. After 24 hours, cells are treated with SK1-I hydrochloride (0, 2.5, 5, 7.5, 10 uM) for 24-48 hours. Cell viability is assessed using the MTT assay or CellTiter-Glo. Apoptosis is measured by Caspase-Glo 3/7 assay (caspase-3/7 activation) or by Western blotting for cleaved CASP3. Autophagy is assessed by LC3-II accumulation (LC3B lipidation) by Western blot and by visualizing GFP-LC3 puncta if using reporter cells. The TP53-dependent nature of cell death is determined by comparing the responses in TP53 wild-type and TP53-null cells. For selectivity confirmation, cells overexpressing SPHK1 or SPHK2 are treated with SK1-I hydrochloride, and S1P production is measured by LC-MS/MS. |
| Animal Protocol |
Animal/Disease Models: Male C57BL/6 mice (24 ± 3.5 g)[3]
Doses: 75 mg/kg Route of Administration: intraperitoneal (ip) injection; once; 24 hrs (hours) prior to baseline MAP measurement Experimental Results: Dramatically lowered baseline mean arterial blood pressure (MAP). An in vivo protocol for SK1-I hydrochloride uses a mouse model to assess effects on blood pressure. Male C57BL/6 mice (24 +/- 3.5 g) are anesthetized, and a catheter is inserted into the carotid artery for continuous mean arterial pressure (MAP) monitoring. SK1-I hydrochloride is dissolved in a suitable vehicle (e.g., 10% DMSO/90% saline) and administered intraperitoneally as a single bolus injection at a dose of 75 mg/kg, 24 hours prior to baseline MAP measurement. Anandamide is then administered intravenously (1 and 10 mg/kg) to assess the effect of SK1-I pre-treatment on anandamide-induced hypotension. Baseline MAP is recorded before anandamide injection. SK1-I pre-treatment significantly reduces baseline MAP and alters the hypotensive response to anandamide, confirming in vivo target engagement of SPHK1. For antitumor efficacy, a xenograft mouse model would be used. Female nude mice are inoculated subcutaneously with HCT116 cells. When tumors reach ~100 mm3, mice are randomized and treated with SK1-I hydrochloride (50-100 mg/kg, i.p., daily) for 14-21 days. Tumor volumes are measured with calipers. |
| ADME/Pharmacokinetics |
Detailed pharmacokinetic data for SK1-I hydrochloride is not available. As a sphingosine analog and competitive SPHK1 inhibitor, it is expected to be well absorbed and distributed to tissues. For in vivo studies, the compound is administered intraperitoneally at a dose of 75 mg/kg. A standard PK study in mice would involve IV (10 mg/kg) and IP (50 mg/kg) administration, followed by serial plasma sampling and LC-MS/MS analysis to determine T1/2, Cmax, AUC, and bioavailability. The compound is lipophilic (sphingosine analog) and is likely to cross cell membranes readily. Metabolism likely involves hepatic CYP450 enzymes and conversion to S1P analogs.
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| Toxicity/Toxicokinetics |
Specific toxicity data for SK1-I hydrochloride is limited. It is described as a selective SPHK1 inhibitor with no activity against SPHK2 or a panel of other protein kinases, suggesting that off-target toxicity may be low. In vitro, SK1-I hydrochloride induces TP53-dependent cell death, indicating that TP53 status may influence the cytotoxic effects of the compound. In vivo, at a dose of 75 mg/kg (i.p.), no acute toxicity was reported in mice. Standard toxicity screening would include in vitro hERG channel inhibition testing, CYP450 enzyme inhibition screening, and a 7-day repeat-dose toxicity study in mice to determine the maximum tolerated dose (MTD).
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| References |
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| Additional Infomation |
SK1-I hydrochloride (BML-258 hydrochloride) is a research-grade chemical and is not approved for clinical use. Its molecular formula is C17H37NO2 with a molecular weight of 277.4 (free base) and higher as hydrochloride salt. It is a sphingosine analog that acts as a selective, competitive, and isozyme-specific inhibitor of sphingosine kinase 1 (SPHK1) with a Ki value of 10 uM. SK1-I hydrochloride shows no activity against SPHK2 or against PKCalpha, PKCdelta, PKA, AKT1, ERK1, EGFR, CDK2, IKKbeta, or CamK2beta. The compound enhances autophagy and has antitumor activity. It is a valuable research tool for studying the sphingolipid signaling pathway, cancer cell survival, and blood pressure regulation.
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| Molecular Formula |
C17H27NO2.CLH
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|---|---|
| Molecular Weight |
313.87
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| Exact Mass |
313.18
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| CAS # |
2366222-05-9
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| Related CAS # |
SK1-I;1072443-89-0
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| PubChem CID |
49793059
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| Appearance |
Colorless to light yellow liquid
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
9
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| Heavy Atom Count |
21
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| Complexity |
260
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| Defined Atom Stereocenter Count |
2
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| SMILES |
CCCCCC1=CC=C(C=C1)/C=C/[C@@H]([C@@H](CO)NC)O.Cl
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| InChi Key |
SGCJOKUPGVFNKS-UUCPMUBFSA-N
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| InChi Code |
InChI=1S/C17H27NO2.ClH/c1-3-4-5-6-14-7-9-15(10-8-14)11-12-17(20)16(13-19)18-2;/h7-12,16-20H,3-6,13H2,1-2H3;1H/b12-11+;/t16-,17+;/m1./s1
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| Chemical Name |
(E,2R,3S)-2-(methylamino)-5-(4-pentylphenyl)pent-4-ene-1,3-diol;hydrochloride
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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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 (796.53 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (6.63 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 20.8 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: ≥ 2.08 mg/mL (6.63 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), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 20.8 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: ≥ 2.08 mg/mL (6.63 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1860 mL | 15.9302 mL | 31.8603 mL | |
| 5 mM | 0.6372 mL | 3.1860 mL | 6.3721 mL | |
| 10 mM | 0.3186 mL | 1.5930 mL | 3.1860 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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