| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| Other Sizes |
| Targets |
The primary target of SPL-334 is S-nitrosoglutathione reductase (GSNOR/ADH5), an enzyme that regulates intracellular levels of S-nitrosoglutathione (GSNO). By inhibiting GSNOR, SPL-334 increases endogenous GSNO levels, thereby amplifying nitric oxide signaling. GSNO acts as a physiological nitric oxide donor and modulates protein S-nitrosylation. SPL-334 shows selectivity for GSNOR over ADH1B, ADH4, and ADH7. This mechanism is relevant to allergic airway inflammation and interstitial lung disease.
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| ln Vitro |
In vitro, SPL-334 inhibits GSNOR with an IC50 of 300 nM against recombinant human GSNOR and a Kd of 2.0 µM. It reduces the production of Th2 cytokines IL-5 and IL-13 and the chemokine CCL11 in airways. In bronchial epithelial cells, SPL-334 inhibits TNF-α-induced IL-6 release with an IC50 of approximately 1 nM. The compound is selective for GSNOR over other alcohol dehydrogenases. It is typically tested at concentrations ranging from 0.1 to 10 µM.
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| ln Vivo |
Reduction in allergic airway relays is caused by SPL-334 (0.1-1 mg/kg; intranasal label; once daily for 7 days; BALB/c junction box labeled DO11.10 CD4+ Th2 xenograft) [1].
In vivo, SPL-334 (0.1-1 mg/kg, intranasal, once daily for 7 days) significantly reduces lymphocyte and eosinophil influx into airways and EPO levels in BALF in a BALB/c mouse model with DO11.10 CD4+ Th2 xenografts. It reduces OVA-specific T cells, eosinophils, peribronchial inflammation, and mucus secretion during allergic airway inflammation. In a bleomycin injury model, SPL-334 prevents weight loss and attenuates lung inflammation and fibrosis. |
| Enzyme Assay |
The GSNOR inhibition assay uses recombinant human GSNOR incubated with GSNO and NADH in reaction buffer. SPL-334 is added at serial dilutions. The reduction of GSNO to GSSG is measured by monitoring NADH oxidation at 340 nm or by HPLC. IC50 values are calculated from dose-response curves. The assay is performed in 96-well or 384-well format for high-throughput screening.
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| Cell Assay |
For cell-based assays, bronchial epithelial cells or Th2 cells are treated with SPL-334. Cytokine production (IL-5, IL-13, IL-6, CCL11) is measured by ELISA. Intracellular GSNO levels are measured by LC-MS or fluorescent probes. Cell viability is assessed using MTT or CellTiter-Glo. For T cell proliferation, CFSE dilution is measured by flow cytometry.
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| Animal Protocol |
Animal/Disease Models: BALB/c recipient mice with DO11.10 CD4+ Th2 xenografts[1]
Doses: 0.1 or 1 mg/kg Route of Administration: Intranasal; one time/day for 7 days Experimental Results: Resulting in a significant decrease in lymphocyte and eosinophil influx into the airways and EPO levels in BALF. Reduces the number of OVA-specific T cells and eosinophils during allergic airway inflammation. Reduces peribronchial inflammation and mucus secretion during airway inflammation. In vivo studies with SPL-334 employ BALB/c mice with DO11.10 CD4+ Th2 xenografts or bleomycin-induced lung injury models. The compound is dissolved in saline and administered intranasally at 0.1-1 mg/kg once daily for 7 days. Key readouts include lymphocyte and eosinophil counts in BALF, cytokine levels, peribronchial inflammation, and mucus secretion. Lung tissue is collected for histological analysis. |
| References |
| Molecular Formula |
C22H15N3O3S2
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|---|---|
| Molecular Weight |
433.5
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| Exact Mass |
433.055
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| CAS # |
688347-51-5
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| PubChem CID |
3651705
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| Appearance |
White to off-white solid
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
694.0±65.0 °C at 760 mmHg
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| Flash Point |
373.5±34.3 °C
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| Vapour Pressure |
0.0±2.3 mmHg at 25°C
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| Index of Refraction |
1.721
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| LogP |
3.85
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
6
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| Heavy Atom Count |
30
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| Complexity |
741
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C(C1C=CC(C(=O)O)=CC=1)N1C(C2SC=CC=2N=C1SCC1C=CC=CC=1C#N)=O
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| InChi Key |
UWWSCLNCHCROLL-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C22H15N3O3S2/c23-11-16-3-1-2-4-17(16)13-30-22-24-18-9-10-29-19(18)20(26)25(22)12-14-5-7-15(8-6-14)21(27)28/h1-10H,12-13H2,(H,27,28)
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| Chemical Name |
4-[[2-[(2-cyanophenyl)methylsulfanyl]-4-oxothieno[3,2-d]pyrimidin-3-yl]methyl]benzoic acid
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| Synonyms |
SPL 334; SPL334; SPL-334
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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: ~50 mg/mL (~115.3 mM; with ultrasonication)
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3068 mL | 11.5340 mL | 23.0681 mL | |
| 5 mM | 0.4614 mL | 2.3068 mL | 4.6136 mL | |
| 10 mM | 0.2307 mL | 1.1534 mL | 2.3068 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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