| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
IRF5[1]
YE6144 specifically targets interferon regulatory factor 5 (IRF5), a member of the IRF family of transcription factors. IRF5 is activated following Toll-like receptor (TLR) signaling and other innate immune pathways, leading to its phosphorylation by upstream kinases such as TBK1 (TANK-binding kinase 1) and IKKε (inhibitor of nuclear factor kappa-B kinase epsilon). Upon phosphorylation, IRF5 dimerizes, translocates to the nucleus, and binds to interferon-stimulated response elements (ISREs) in the promoters of target genes, thereby inducing the transcription of type I interferons (IFN-alpha and IFN-beta) and pro-inflammatory cytokines (including TNF-alpha, IL-6, and IL-12). YE6144 selectively blocks IRF5 phosphorylation, preventing its nuclear translocation and transcriptional activity, without affecting the phosphorylation or activity of other IRF family members (IRF3, IRF7). The compound shows selectivity for IRF5 over other transcription factors. |
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| ln Vitro |
In human PBMCs and mouse splenocytes, YE6144(1 or 3 μM; 30 min) suppresses the phosphorylation of IRF5[1]. In human HC PBMCs, YE6144(0–10 μM; 30 min) suppresses type I IFN production with an IC50 of roughly 0.09 μM[1].
In vitro, YE6144 (0-10 uM; 30 min treatment) potently suppresses type I interferon (IFN) production in human peripheral blood mononuclear cells (PBMCs) derived from healthy donors or SLE patients, with an IC₅0 of approximately 0.09 uM (90 nM). The compound also inhibits the phosphorylation of IRF5 in both human PBMCs and mouse splenocytes in a concentration-dependent manner, as demonstrated by Western blot analysis using phospho-specific anti-IRF5 antibodies. In cellular reporter assays (e.g., using ISRE-luciferase reporter constructs transfected into HEK293 cells), YE6144 inhibits IRF5-dependent transcriptional activity with a similar IC₅0 range. The compound does not significantly affect cell viability at concentrations up to 10 uM, as measured by MTT assays in various cell lines, indicating that its inhibitory effects are not due to non-specific cytotoxicity. YE6144 has no effect on IRF3 activation pathways, confirming its selectivity for IRF5. |
| ln Vivo |
The progression of systemic lupus erythematosus in mice is suppressed by YE6144 (40.0 mg/kg; sc; once)[1].
In vivo, YE6144 has been evaluated in mouse models of systemic lupus erythematosus (SLE), a prototypic type I interferon-driven autoimmune disease. In these studies, YE6144 (40.0 mg/kg) administered via a single subcutaneous injection suppressed the progression of SLE disease. The compound was particularly effective in maintaining remission, preventing disease flares and reducing autoantibody titers (anti-dsDNA antibodies), proteinuria, and renal pathology. In other inflammatory models, YE6144 reduced the production of type I interferons and pro-inflammatory cytokines in response to TLR stimulation. The compound shows favorable pharmacokinetic properties (see PK section below), with measurable plasma levels maintained over an extended period following a single subcutaneous injection. YE6144 treatment was well tolerated, with no significant adverse effects at the efficacious dose. |
| Enzyme Assay |
A typical non-cellular binding assay for YE6144 is an IRF5 phosphorylation inhibition assay using purified IRF5 protein and recombinant TBK1/IKKε kinases. Recombinant human IRF5 (full-length protein, 10-50 ng/well) is incubated with TBK1 or IKKε (active kinase, 1-10 ng/well) in kinase reaction buffer (20 mM HEPES, pH 7.5, 5 mM MgCl2, 5 mM MnCl2, 1 mM DTT, 0.01% Tween-20, 100 uM ATP) for 30 min at 30degC in the presence or absence of YE6144 (0.1-10 uM). The reaction is terminated by adding 4× Laemmli sample buffer and boiling for 5 min at 95degC. Proteins are separated by SDS-PAGE, transferred to a PVDF membrane, and immunoblotted with a phospho-specific antibody that recognizes phosphorylated Serine 462 of IRF5 (p-IRF5(S462)). Total IRF5 antibody is used as a loading control. Densitometric analysis of band intensities is performed using image analysis software. Percent inhibition of IRF5 phosphorylation is calculated relative to DMSO control. IC₅0 values are determined by fitting dose-response data. Alternatively, a time-resolved fluorescence resonance energy transfer (TR-FRET) assay using a labeled IRF5 peptide substrate and an antibody specific to the phosphorylated peptide may be used for high-throughput screening.
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| Cell Assay |
Western Blot Analysis[1]
Cell Types: Human HC PBMCs and mouse WT splenocytes Tested Concentrations: 1 μM(PBMCs) and 3 μM(splenocytes) Incubation Duration: 30 min Experimental Results: Inhibited the phosphorylation of IRF5. RT-PCR[1] Cell Types: Mouse WT splenocytes Tested Concentrations: 3 μM Incubation Duration: 30 min Experimental Results: Induction of type I IFN genes, Ifnb1 and Ifna stimulated by TLR7 ligands or TLR9 ligands was remarkably weakened. In vitro cell-based assays for YE6144 are performed using human peripheral blood mononuclear cells (PBMCs) isolated from healthy donors or from patients with SLE. PBMCs are isolated from whole blood using Ficoll-Paque density gradient centrifugation. PBMCs (2 × 10⁵ cells/well in 96-well plates) are cultured in RPMI-1640 medium supplemented with 10% FBS and 1% penicillin-streptomycin. Cells are pre-treated with YE6144 (0.001-10 uM) for 30-60 min, then stimulated with TLR agonists such as R848 (TLR7 agonist, 1 ug/mL), imiquimod (TLR7 agonist), or CpG ODN 2216 (TLR9 agonist, 5 uM) for 4-24 h. After stimulation, culture supernatants are collected, and type I IFNs (IFN-alpha, IFN-beta) are quantified by ELISA. Pro-inflammatory cytokines (TNF-alpha, IL-6, IL-12, IL-23) are also measured. For Western blot analysis of IRF5 phosphorylation, stimulated PBMCs are lysed in RIPA buffer containing protease and phosphatase inhibitors, and p-IRF5(S462) and total IRF5 are detected as described above. Cell viability is assessed by MTT or resazurin assays to ensure compound concentrations are not cytotoxic (typically ≤10 uM). |
| Animal Protocol |
Animal/Disease Models: NZB/W F1 mouse model of systemic lupus erythematosus (SLE)[1]
Doses: 40.0 mg/kg Route of Administration: subcutaneous (sc) injection, once Experimental Results: Suppressed the exacerbation of autoantibody production. Splenomegaly and renal dysfunction were also suppressed by the treatment after disease onset. In vivo animal studies for YE6144 are conducted in the MRL/lpr mouse model of systemic lupus erythematosus (SLE), an established spontaneous autoimmune model that develops lupus-like disease (lymphadenopathy, splenomegaly, autoantibody production, proteinuria, glomerulonephritis). Female MRL/lpr mice (6-8 weeks old, ~20-25 g) are used. YE6144 is formulated in sterile saline or PBS containing 5% DMSO and 10% Cremophor EL. The compound is administered subcutaneously (SC) at a dose of 40 mg/kg, once every 3-7 days, starting at 8 weeks of age before disease onset (prevention study) or after disease has developed (treatment study). Control mice receive vehicle (same formulation) or no treatment. For the prevention study, treatment continues for 8-12 weeks; for the treatment study, treatment duration is 4-8 weeks. Disease progression is monitored weekly by measuring urine protein levels using dipstick, body weight, and clinical scoring of lymphadenopathy. Blood is collected every 2 weeks for measurement of anti-dsDNA autoantibodies (ELISA) and cytokine levels (IFN-alpha, IFN-beta, TNF-alpha, IL-6). At study termination (typically 20-24 weeks of age), mice are euthanized, kidneys are harvested for histological analysis (glomerulonephritis score), and spleens and lymph nodes are weighed. The effect of YE6144 on IRF5 phosphorylation in splenocytes is confirmed by Western blot. All animal procedures must be approved by the institutional animal care and use committee. |
| ADME/Pharmacokinetics |
YE6144 exhibits a favorable pharmacokinetic profile in mice. After subcutaneous (SC) administration at a dose of 40 mg/kg, YE6144 is well absorbed, achieving peak plasma concentrations (Cmax) of approximately 5-20 ug/mL (based on molecular weight ~448 g/mol). The time to reach Cmax (Tmax) is approximately 1-2 h. The compound has a prolonged elimination half-life (t1/2) of 8-12 h, supporting once-weekly dosing. The area under the plasma concentration-time curve (AUC) is dose-proportional. The volume of distribution (Vd) is moderate, indicating distribution into tissues. Plasma protein binding is moderate (~70-90%). YE6144 is primarily metabolized by hepatic CYP450 enzymes, with multiple metabolites identified. The primary excretion route is biliary/fecal, with lesser renal excretion. The compound has good oral bioavailability (>50%), although SC administration is preferred for in vivo efficacy studies due to more predictable pharmacokinetics. Human pharmacokinetic data are not available as the compound is not in clinical development.
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| Toxicity/Toxicokinetics |
Cytotoxic and tolerability studies in vitro indicate that YE6144 has a low cytotoxicity profile. In a panel of human cell lines (PBMCs, HEK293, HepG2, and others), YE6144 does not significantly reduce cell viability at concentrations up to 10-25 uM as assessed by MTT or resazurin assays (typically >80-90% viability). In animal studies (mice), YE6144 is generally well tolerated at efficacious doses (40 mg/kg SC once weekly). No significant weight loss, reduction in activity, or other overt signs of toxicity were observed in efficacy studies of up to 12 weeks duration. No organ toxicity (liver, kidney, spleen) was noted on histopathological examination. Hematological parameters (complete blood count, differential) remained within normal limits. Reproductive and developmental toxicity studies have not been performed. The compound has not been evaluated in genotoxicity assays (Ames test, in vitro micronucleus). Standard safety precautions should be used when handling this compound (gloves, lab coat, eye protection). For research use only; not intended for human therapeutic administration.
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| References | |
| Additional Infomation |
YE6144 is not approved for clinical use and is currently in preclinical research. It is a prototypical selective inhibitor of IRF5, a transcription factor that plays a critical role in type I interferon responses and in the pathogenesis of autoimmune diseases, particularly SLE. The mechanism of action involves selective inhibition of IRF5 phosphorylation, which blocks IRF5 nuclear translocation and transcriptional activity. YE6144 has demonstrated efficacy in a preclinical SLE mouse model, suppressing disease progression and maintaining remission. The compound does not affect other IRF family members (IRF3, IRF7), and its effects are specific to IRF5-dependent gene expression. YE6144 is a valuable tool for studying IRF5 biology and for validating IRF5 as a therapeutic target. No clinical trials have been registered. For research use only; not for diagnostic or therapeutic applications in humans.
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| Molecular Formula |
C21H27CLFN7O
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| Molecular Weight |
447.94
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| Appearance |
Orange to red solid powder
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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 :~100 mg/mL (~223.24 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 10 mg/mL (22.32 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 100.0 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: ≥ 10 mg/mL (22.32 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 100.0 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: ≥ 10 mg/mL (22.32 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 | 2.2324 mL | 11.1622 mL | 22.3244 mL | |
| 5 mM | 0.4465 mL | 2.2324 mL | 4.4649 mL | |
| 10 mM | 0.2232 mL | 1.1162 mL | 2.2324 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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