| Size | Price | Stock | Qty |
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| 500mg |
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| 1g |
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| Other Sizes |
Purity: ≥98%
| Targets |
Receptor interacting protein 1 (RIP1)
GNE684 targets receptor-interacting protein 1 (RIP1), a serine/threonine-protein kinase that plays a central role in the regulation of cell death and inflammation. RIP1 is a key upstream activator of necroptosis (a form of programmed necrosis) and also regulates NF-κB-mediated inflammatory signaling. Inhibition of RIP1 kinase activity blocks necroptosis and reduces inflammatory cytokine production. GNE684 potently inhibits human RIP1 with an apparent Ki of 21 nM. The compound's species selectivity (higher potency for human vs. mouse and rat RIP1) is important for interpreting preclinical studies. |
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| ln Vitro |
In multiple human and animal cell lines, GNE684 (20 μM; 20 hours) efficiently suppresses RIP1 kinase-driven cell death[1]. GNE684 (20 μM; 0–60 minutes) inhibits RIP1 autophosphorylation generated by TBZ (2 μM BV6, 20 ng/ml TNF, 20 μM zVAD), as well as RIP3 autophosphorylation and RIP3-mediated phosphorylation of MLKL[1].
GNE684 demonstrates potent inhibition of human RIP1 with an apparent Ki of 21 nM. The compound shows lower potency against mouse RIP1 (189 nM) and rat RIP1 (691 nM). In cellular assays, GNE684 blocks RIP1 kinase activity, preventing necroptosis induced by TNF-α and other stimuli. The compound also inhibits RIP1-dependent inflammatory signaling, including NF-κB activation and cytokine production. GNE684's in vitro activity has been characterized in various cell lines, confirming its mechanism as a selective RIP1 kinase inhibitor. |
| ln Vivo |
Additionally, in the KPP or KPR (LSL-Kras G12D/+; p16/p19 fl/wt; Trp53 R270H/wt; Pdx1-cre) PDAC models, GNE684 had no effect on overall survival or tumor growth[1]. Intestinal epithelial cells (IECs) that lack NEMO are the source of colitis and ileitis, which are inhibited by GNE684 (50 mg/kg; po twice daily)[1].
In vivo, GNE684 has been shown to inhibit colitis and ileitis caused by NEMO deficiency in intestinal epithelial cells in mouse models. Oral administration of GNE684 at doses such as 50 mg/kg twice daily reduces intestinal inflammation and tissue damage in these models. The compound's efficacy in blocking RIP1-mediated inflammation supports the therapeutic potential of RIP1 inhibition for inflammatory bowel disease and other immune-mediated conditions. In vivo pharmacodynamic studies confirm target engagement and suppression of RIP1-dependent inflammatory pathways. |
| Enzyme Assay |
In vitro enzyme/receptor binding assays for GNE684 measure inhibition of RIP1 kinase activity using recombinant RIP1 enzyme and a suitable peptide substrate. Kinase activity is assessed by measuring the incorporation of ³²P from [γ-³²P]ATP into the substrate, or by using a luminescent ADP detection assay (e.g., ADP-Glo). IC50 or Ki values are determined from dose-response curves. Selectivity is assessed by screening GNE684 against a panel of kinases to confirm specificity for RIP1 over other kinases. Binding affinity (Kd) can be measured by surface plasmon resonance (SPR) or by competition binding assays using a fluorescently labeled RIP1 inhibitor.
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| Cell Assay |
Cell Viability Assay[1]
Cell Types: L929 cells, Jurkat cells, MEFs Tested Concentrations: 20 μM Incubation Duration: 20 hrs (hours) Experimental Results: Inhibited RIP1 kinase driven cell death effectively in several human and mouse cell lines. Western Blot Analysis[1] Cell Types: HT-29 cells, J774A.1 cells Tested Concentrations: 0 μM, 20 μM Incubation Duration: 0 minute, 15 minutes, 60 minutes Experimental Results: Disrupted TBZ (2 μM BV6, 20 ng/ml TNF, 20μM zVAD)-induced RIP1 autophosphorylation, interactions between RIP1 and RIP3, RIP3 autophosphorylation, and phosphorylation of MLKL by RIP3. In vitro cellular assays for GNE684 are performed in cell lines to assess inhibition of RIP1-dependent necroptosis and inflammatory signaling. Necroptosis is induced by treatment with TNF-α, a SMAC mimetic, and a pan-caspase inhibitor (TSZ), and cell death is measured by propidium iodide uptake, LDH release, or CellTiter-Glo. GNE684's ability to block necroptosis is determined by comparing cell viability in treated versus untreated cells. RIP1-dependent NF-κB activation is assessed by measuring IκBα phosphorylation and degradation, or by reporter gene assays. Cytokine production (e.g., TNF-α, IL-6, IL-1β) is measured by ELISA. |
| Animal Protocol |
Animal/Disease Models: Nemofl/fl Villin.creERT2 mice (NEMO IEC-KO)[1]
Doses: 50 mg/kg Route of Administration: Oral administration; twice (two times) daily; from days 2–6 treated with tamoxifen Experimental Results: Almost completely protected the NEMO-deficient intestines from colitis and ileitis. In vivo animal studies with GNE684 are conducted in mouse models of inflammation and necroptosis. The NEMO-deficient intestinal epithelial cell model is used to study colitis and ileitis, with GNE684 administered orally at 50 mg/kg twice daily. Disease severity is assessed by histology, inflammatory cytokine levels, and clinical scores. Other models of RIP1-dependent inflammation may include TNF-induced systemic inflammatory response syndrome (SIRS) and models of ischemia-reperfusion injury. Pharmacodynamic endpoints include assessment of RIP1 activity, necroptosis markers (e.g., phosphorylated MLKL), and inflammatory cytokine levels in tissues. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties of GNE684 have been characterized to support in vivo efficacy studies. Following oral administration at 50 mg/kg, the compound achieves therapeutic plasma concentrations sufficient for RIP1 inhibition. The compound's half-life, clearance, volume of distribution, and oral bioavailability have been determined in preclinical species. GNE684 shows good oral absorption and reasonable plasma exposure. Metabolism occurs via hepatic cytochrome P450 enzymes. The PK profile supports twice-daily dosing in preclinical efficacy studies.
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| Toxicity/Toxicokinetics |
Toxicological data for GNE684 are primarily derived from preclinical efficacy studies in mouse models. At efficacious doses (e.g., 50 mg/kg twice daily), the compound appears to be tolerated with no severe adverse effects reported in published literature. However, comprehensive toxicology studies including genotoxicity, safety pharmacology, and repeated-dose toxicity in rodent and non-rodent species would be required for clinical development. As a RIP1 inhibitor, potential on-target toxicities could include effects on immune function and tissue homeostasis, given the role of RIP1 in regulating cell death and inflammation.
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| References | |
| Additional Infomation |
The kinase RIP1 participates in multiple signaling pathways, regulates inflammatory responses, and can induce apoptosis and necroptosis. Its kinase activity is associated with various inflammatory diseases, neurodegenerative diseases, and neoplastic diseases. This study investigated the effects of gene knock-in expression of the catalytically inactive RIP1 D138N mutant in mice and pharmacological inhibition of RIP1 using the potent murine inhibitor GNE684. Results showed that RIP1 inhibition alleviated collagen antibody-induced arthritis and prevented skin inflammation caused by Sharpin gene mutations and colitis caused by Nemo gene deletion in intestinal epithelial cells. Conversely, RIP1 inhibition had no effect on tumor growth or survival in a mutant Kras-driven pancreatic tumor model, nor did it reduce lung metastases in a B16 melanoma model. In summary, our data highlight the role of RIP1 kinase activity in certain inflammatory disease models, but raise questions about its association with tumor progression and metastasis.
GNE684 is a potent inhibitor of RIP1 kinase with an apparent Ki of 21 nM for human RIP1. It shows species selectivity with lower potency against mouse (189 nM) and rat (691 nM) RIP1. GNE684 has demonstrated efficacy in inhibiting colitis and ileitis in mouse models. It is a research tool for studying RIP1 in necroptosis and inflammation. No clinical development or regulatory approvals have been reported. The compound is available for laboratory research purposes. |
| Molecular Formula |
C23H24N6O3
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|---|---|
| Molecular Weight |
432.475064277649
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| Exact Mass |
432.190988
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| Elemental Analysis |
C, 63.88; H, 5.59; N, 19.43; O, 11.10
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| CAS # |
2438637-64-8
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| PubChem CID |
138377384
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
2.4
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| SMILES |
N1=C(C(N[C@H]2C(=O)N(C)C3C=NC(OC)=CC=3CC2)=O)N=C2CC[C@H](C3=CC=CC=C3)N12
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| InChi Key |
JXFYROJRZJPKTQ-IRXDYDNUSA-N
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| InChi Code |
InChI=1S/C23H24N6O3/c1-28-18-13-24-20(32-2)12-15(18)8-9-16(23(28)31)25-22(30)21-26-19-11-10-17(29(19)27-21)14-6-4-3-5-7-14/h3-7,12-13,16-17H,8-11H2,1-2H3,(H,25,30)/t16-,17-/m0/s1
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| Chemical Name |
(5S)-N-[(3S)-7-methoxy-1-methyl-2-oxo-4,5-dihydro-3H-pyrido[3,4-b]azepin-3-yl]-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide
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| Synonyms |
GNE-684; GNE 684; GNE684; 2438637-64-8; CHEMBL5208359; (5S)-N-[(3S)-7-Methoxy-1-methyl-2-oxo-4,5-dihydro-3H-pyrido[3,4-b]azepin-3-yl]-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide; (5S)-N-[(3S)-7-methoxy-1-methyl-2-oxo-2,3,4,5-tetrahydro-1H-pyrido[3,4-b]azepin-3-yl]-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide; rel-(S)-N-((S)-7-Methoxy-1-methyl-2-oxo-2,3,4,5-tetrahydro-1H-pyrido[3,4-b]azepin-3-yl)-5-phenyl-6,7-dihydro-5H-pyrrolo[1,2-b][1,2,4]triazole-2-carboxamide; SCHEMBL25798914; GTPL13107; GNE684
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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.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.3122 mL | 11.5612 mL | 23.1225 mL | |
| 5 mM | 0.4624 mL | 2.3122 mL | 4.6245 mL | |
| 10 mM | 0.2312 mL | 1.1561 mL | 2.3122 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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