SH514
Based on 1 Customer Validation
SH514 is an orally active IRF4 inhibitor (IC50 = 2.63 μM). SH514 binds to the IRF4-DBD domain, thereby inhibiting the interaction of IRF4 protein with DNA (KD = 1.28 μM). SH514 can inhibit the proliferation of IRF4-high-expressing NCI-H929 and MM.1R cells, and displays no cytotoxicity for normal cells. SH514 significantly downregulates the expression of IRF4 downstream target genes concentration-dependently. SH514 inhibits the expression of cell cycle-related proteins CDC2, Cyclin B1, Cyclin D1, Cyclin E1, and CMYC in Multiple Myeloma cells. SH514 can induce DNA damage and increase the expression of γH2AX. SH514 effectively inhibits the proliferation of multiple myeloma tumors .
For research use only. We do not sell to patients.
- Purity : 99.94%
- CAS No.: 3099543-90-2
- Formula: C32H38N2O4
- Molecular Weight:514.66
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Protocols
- Purity & Documentation
- References
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Help & FAQs
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Biological Activity
Description
In Vitro
SH514 (Compound 41) (0-50 μM, 72 h) targets IRF4 at the cellular level, can selectively inhibit the proliferation of MM cell lines with high IRF4 expression, and displays no cytotoxicity for normal cells[1].
SH514 (100 μM, 1 h) can target IRF4 and reduce its thermal stability, while having almost no effect on the stability of IRF3, a member of the IRF family[1].
SH514 (0.125-1 μM, 24 h) significantly downregulates the expression of IRF4 downstream target genes in NCI-H929 and MM.1R cells, and the inhibitory effect was concentration-dependent in MM cells[1].
SH514 (0-0.5 μM, 24 h) leads to a significant increase in the proportion of cells in the G1 phase and a decrease in cells in the S and G2 phases in NCI-H929 cells[1].
SH514 (0.1-1 μM, 24 h) suppresses the expression of p-AKT, p-ERK, CDC2, Cyclin B1, Cyclin D1, Cyclin E1, and CMYC, and increase γH2AX in NCI-H929 cells and MM.1R cells, suggesting that SH514 could induce DNA damage[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:NCI-H929, MM.1R, RPMI-8226, HAF, HACAT
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Concentration:0-50 μM
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Incubation Time:72 h
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Result:Demonstrated potent inhibition against IRF4-high expressing cells NCI-H929 (IC50 = 0.08 μM) and MM.1R (IC50 = 0.11 μM).
Showed weaker inhibitory effects on IRF4-low expressing cells RPMI-8226 (IC50 = 1.19 μM).
Exhibited no toxicity towards normal cells, HAF and HACAT (IC50 > 100 μM).
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Cell Line:NCI-H929, MM.1R
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Concentration:0.125 μM, 0.25 μM, 0.5 μM, 1 μM
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Incubation Time:24 h
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Result:Concentration-dependently downregulated the mRNA expression of IRF4 downstream target genes, including CCNC, CANX, E2F5, CMYC, HK2, and Blimp1.
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Cell Line:NCI-H929
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Concentration:0.125 μM, 0.25 μM, 0.5 μM
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Incubation Time:24 h
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Result:Induced cell cycle arrest at the G1 phase in NCI–H929 cells in a concentration-dependent manner.
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Cell Line:NCI-H929, MM.1R
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Concentration:0.1 μM, 0.3 μM, 0.5 μM, 1 μM
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Incubation Time:24 h
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Result:Concentration gradients inhibited the expression of the cell cycle-related proteins CDC2, Cyclin B1, Cyclin D1, Cyclin E1, and CMYC in NCI–H929 cells and MM.1R cells.
Parmacokinetics
| Species | Dose | Route | Tmax | Cmax | AUC0-t | AUC0-∞ | Vz | CL | T1/2 | F |
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| Mice | 1 mg/kg | i.v. | 0.08 h | 174 ng/mL | 81 ng·h/mL | 86 ng·h/mL | 20807 mL/kg | 11818 mL/min/kg | 1.25 h | / |
| Mice | 10 mg/kg | p.o. | 0.53 h | 82 ng/mL | 285 ng·h/mL | 463 ng·h/mL | / | / | 6.71 h | 53.85 % |
In Vivo
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:1 × 107 of H929 cells were injected subcutaneously into BALB/c nude mice by mixing 1:1 with matrix gel[1].
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Dosage:12.5 mg/kg, 25 mg/kg
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Administration:Oral gavage, once daily for 34 days
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Result:Significantly inhibited the tumor volume in mice in a dose-dependent manner.
The body weight of mice showed an overall increasing trend, indicating that the compound had no toxic effect on mice.
Significantly reduced the expression of Ki67.
Effectively inhibited the proliferation of MM tumors in mice without causing toxic effects.
Chemical Information
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CAS No. 3099543-90-2
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Appearance Solid
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Molecular Weight 514.66
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Formula C32H38N2O4
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Color White to off-white
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SMILES
C[C@]12CC[C@@]3([H])[C@@]([H])(C(C=C4C(C)(C(C(C#N)=C[C@@]43C)=O)C)=O)[C@]1([H])CC[C@@H]2[C@@H](C(NC5=CC=C(C=C5)OC)=O)C
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Solvent & Solubility
In Vitro:
DMSO : 100 mg/mL (194.30 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
In Vivo:
Select the appropriate dissolution method based on your experimental animal and administration route.
- For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
- To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for In Vivo experiments, it is recommended to prepare freshly and use it on the same day.
- The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
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Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
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%DMSO +
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
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%+
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+%Tween-80 + +
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%Saline +
The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Working solution concentration: 0.22 mg/mL
Method for preparing stock solution: mg drug dissolved in μL DMSO. Stock solution concentration: mg/mL.
1. Take μL DMSO stock solution;
2. Add μL .
μL , mix evenly;
3. Then add μL Tween 80, mix evenly;
4. Then add μL
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
Protocols
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RNA extraction experimental
By lysing cells, releasing RNA, and removing impurities such as proteins and DNA, high-purity RNA products are finally obtained. The commonly used traditional method is the guanidine isothiocyanate/phenol/chloroform method (Trizol), which is suitable for a variety of animal materials including animal tissues, microorganisms, cultured cells, etc., and most plant materials.
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Cell Cytotoxicity Assay
Cytotoxicity assays are usually based on the assessment of cell membrane damage, which can also be indirectly detected by measuring cell viability. Detection methods include MTT assay, CKK-8 assay, LDH assay and ATP assay, etc.
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Mammalian live/dead viability and cytotoxicity staining
Live/dead viability and cytotoxicity staining assays are based on the simultaneous detection of intracellular esterase activity in metabolically active (viable) cells and membrane integrity loss in non-viable cells. In commonly used dual-staining approaches, membrane-permeant fluorogenic substrates are converted by intracellular esterases into fluorescent products in live cells, while impermeant DNA-binding dyes selectively enter cells with compromised plasma membranes and label nucleic acids in dead or dying cells, enabling discrimination between viable and non-viable populations by fluorescence microscopy or flow cytometry.
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Flow cytometric DNA-content cell-cycle staining
Flow cytometric DNA-content cell-cycle staining measures the fluorescence intensity of DNA-bound fluorochromes in single cells or nuclei to estimate DNA content distributions, allowing assignment of populations to G0/G1, S, and G2/M phases by DNA histogram deconvolution. Propidium iodide (PI) intercalates into DNA, and PI fluorescence is proportional to cellular DNA content when staining is performed under conditions that make DNA accessible and minimize non-DNA signal. Cells with G2/M DNA content are expected to show approximately twice the fluorescence intensity of G0/G1 cells, while S-phase cells occupy intermediate fluorescence values. PI-based DNA-content analysis can also detect cells with fractional DNA content, often reported as sub-G1, when DNA fragmentation and extraction during staining reduce retained DNA signal in apoptotic cells. DAPI is an alternative DNA fluorochrome for univariate DNA-content analysis, while bivariate approaches combining DNA content with proliferation
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BrdU Incorporation Assay
Bromodeoxyuridine (BrdU) incorporation assay is based on the principle that BrdU, a thymidine analog, is incorporated into newly synthesized DNA during the S phase of the cell cycle, thereby serving as a marker of DNA replication and cellular proliferation. Incorporated BrdU can be detected using anti-BrdU antibodies following DNA denaturation, enabling visualization or quantification of proliferating cells through immunochemical detection methods such as immunofluorescence or immunohistochemistry.
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Protocol for Cell Cycle
Cell-cycle analysis by flow cytometry measures DNA content in single cells to estimate the fraction of cells in G0/G1, S, and G2/M phases. Propidium iodide intercalates into DNA, and after RNA removal with RNase, fluorescence intensity reflects cellular DNA content: 2N cells are assigned to G0/G1, cells between 2N and 4N to S phase, and 4N cells to G2/M. DNA-content analysis alone cannot reliably separate G0 from G1 or G2 from M. Ki-67 can distinguish quiescent G0 cells from cycling cells, EdU or BrdU incorporation marks active DNA synthesis in S phase, and phospho-histone H3 staining identifies mitotic cells within the 4N population.
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Genotoxicity/Mutagenicity Study
The bacterial reverse mutation assay detects point mutations that restore amino-acid prototrophy in auxotrophic Salmonella typhimurium or Escherichia coli tester strains; after exposure to a test article, mutagenic activity is read out as an increased number of revertant colonies on minimal agar compared with the vehicle control. The assay uses tester strains with different mutation targets so that base-substitution and frameshift mutagens can be detected, and testing is performed with and without exogenous mammalian metabolic activation because some chemicals require biotransformation to become mutagenic.
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Cell Viability Determination by MTT Colorimetric Assay
The following protocol uses the MTT colorimetric assay as a classic literature-established method for assessing cell viability/metabolic activity in cultured mammalian cells. MTT[3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] is reduced by metabolically active cells to a colored formazan product; the amount of formazan is quantified spectrophotometrically and provides an indirect measure of metabolically active viable cells. Importantly, MTT reduction reflects cellular oxidoreductase/metabolic activity rather than an absolute direct count of living cells, so changes in cellular metabolism can alter the signal independently of cell number.
Purity & Documentation
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Data Sheet (279 KB)
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SDS (251 KB)
- English - EN (251 KB)
- Français - FR (251 KB)
- Deutsch - DE (251 KB)
- Norwegian - NO (251 KB)
- Español - ES (251 KB)
- Swedish - SV (251 KB)
- Italian - IT (251 KB)
- Korean - KR (251 KB)
- Portuguese - PT (251 KB)
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Handling Instructions (2659 KB)
References
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
|---|---|---|---|---|---|
| DMSO | 1 mM | 1.9430 mL | 9.7152 mL | 19.4303 mL | 48.5758 mL |
| 5 mM | 0.3886 mL | 1.9430 mL | 3.8861 mL | 9.7152 mL | |
| 10 mM | 0.1943 mL | 0.9715 mL | 1.9430 mL | 4.8576 mL | |
| 15 mM | 0.1295 mL | 0.6477 mL | 1.2954 mL | 3.2384 mL | |
| 20 mM | 0.0972 mL | 0.4858 mL | 0.9715 mL | 2.4288 mL | |
| 25 mM | 0.0777 mL | 0.3886 mL | 0.7772 mL | 1.9430 mL | |
| 30 mM | 0.0648 mL | 0.3238 mL | 0.6477 mL | 1.6192 mL | |
| 40 mM | 0.0486 mL | 0.2429 mL | 0.4858 mL | 1.2144 mL | |
| 50 mM | 0.0389 mL | 0.1943 mL | 0.3886 mL | 0.9715 mL | |
| 60 mM | 0.0324 mL | 0.1619 mL | 0.3238 mL | 0.8096 mL | |
| 80 mM | 0.0243 mL | 0.1214 mL | 0.2429 mL | 0.6072 mL | |
| 100 mM | 0.0194 mL | 0.0972 mL | 0.1943 mL | 0.4858 mL |
SH514 Related Classifications
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