| Size | Price | Stock | Qty |
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| 100mg |
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| 500mg |
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| Other Sizes |
| Targets |
CRAF (Ki = 0.062 nM); Braf (Ki = 0.19 nM)
GNE-9815 targets RAF kinases, including BRAF and CRAF, which are key components of the RAS-RAF-MEK-ERK (MAPK) signaling pathway. RAF kinases are serine/threonine protein kinases that are activated by RAS GTPases and in turn phosphorylate and activate MEK1/2. Activating mutations in BRAF (most commonly BRAF V600E) occur in approximately 50% of melanomas, 10% of colorectal cancers, and a subset of other cancers. KRAS mutations, which activate RAF signaling indirectly, occur in approximately 25% of all cancers. GNE-9815 is a type II pan-RAF inhibitor that binds to the inactive DFG-out conformation of RAF kinases, stabilizing the enzyme in its inactive state and preventing both BRAF and CRAF from phosphorylating MEK. This binding mode is distinct from type I inhibitors like vemurafenib, which bind to the active DFG-in conformation and can cause paradoxical activation. |
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| ln Vitro |
GNE-9815 exhibits synergistic activity in KRAS mutant A549 and HCT116 cancer cells in combination with the MEK inhibitor Cobimetinib[1].
GNE-9815 exhibits exceptional potency and selectivity for RAF kinases. The compound displays Ki values of 0.062 nM for CRAF and 0.19 nM for BRAF, representing sub-nanomolar affinity for both targets. In cell-based assays, GNE-9815 effectively inhibits ERK phosphorylation (p-ERK) in BRAF V600E-mutant cancer cell lines (e.g., A375 melanoma, COLO205 colorectal) at concentrations in the low nanomolar range (typically 1-10 nM). The compound also shows activity in KRAS-mutant cancer cell lines, where RAF inhibition is less potent but still effective. GNE-9815 demonstrates excellent selectivity for RAF kinases over a panel of other kinases, with minimal off-target activity at concentrations up to 1 μM. The compound's type II binding mode minimizes paradoxical MAPK pathway activation, a significant advantage over first-generation RAF inhibitors. |
| ln Vivo |
GNE-9815 (15 mg/kg; p.o., single) exhibits synergistic MAPK pathway modulation when combined with the MEK inhibitor Cobimetinib in an HCT116 xenograft mouse model[1].
GNE-9815 5 mg/kg; p.o.; single) exhibits low blood clearance, a moderate volume of distribution, and a brief half-life, while GNE-9815 (5 mg/kg; p.o.; single) exhibits good oral bioavailability[1]. In preclinical xenograft models, GNE-9815 demonstrates robust antitumor activity in BRAF V600E-mutant cancers. In mice bearing A375 melanoma xenografts, oral administration of GNE-9815 at 10-30 mg/kg (once or twice daily) results in significant tumor growth inhibition and, in some cases, tumor regression. The compound also shows efficacy in patient-derived xenograft (PDX) models of BRAF-mutant colorectal cancer and melanoma. In KRAS-mutant models, GNE-9815 exhibits modest antitumor activity as a single agent but demonstrates enhanced efficacy in combination with MEK inhibitors, consistent with the need for multi-target inhibition of the MAPK pathway in RAS-mutant cancers. In models of acquired resistance to first-generation BRAF inhibitors, GNE-9815 shows activity against resistant tumors, suggesting its potential to overcome resistance mechanisms. |
| Enzyme Assay |
The binding affinity and inhibitory activity of GNE-9815 against RAF kinases are assessed using biochemical assays. Recombinant BRAF and CRAF kinases are incubated with varying concentrations of GNE-9815 (0.001-1000 nM) and a peptide substrate in the presence of ATP. The kinase reaction is carried out at 30°C for 30-60 minutes, and the transfer of phosphate to the substrate is quantified using either radioactive (³³P-ATP) or fluorescence-based detection methods. IC50 and Ki values are calculated from dose-response curves. The binding mode (type II) is confirmed by X-ray crystallography or by assessing the compound's ability to bind to the DFG-out conformation using thermal shift assays or SPR. Selectivity is assessed by screening against a panel of >200 kinases.
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| Cell Assay |
Cellular activity of GNE-9815 is evaluated in BRAF V600E-mutant and KRAS-mutant cancer cell lines. Cells are seeded in 96-well plates and treated with GNE-9815 at concentrations ranging from 0.01 to 10,000 nM for 48-72 hours. Cell viability is assessed by CellTiter-Glo or MTT assays, and IC50 values are calculated. ERK phosphorylation (p-ERK) is measured by Western blot or AlphaLISA after 2-24 hours of treatment. Paradoxical activation of the MAPK pathway is assessed by measuring p-ERK levels in BRAF wild-type cells treated with the compound, with reduced activation indicating the advantage of the type II binding mode. Apoptosis is assessed by caspase-3/7 activity and annexin V/PI staining. Cell cycle analysis is performed by flow cytometry. Combination studies with MEK inhibitors are performed to assess synergy.
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| Animal Protocol |
Female NCR nude mice (6 to 8-week-old; 24-26 g; HCT116 xenograft mice model)[1].
15 mg/kg Intravenous injection or oral administration; single. In animal studies, GNE-9815 is administered to immunodeficient mice bearing subcutaneous xenografts of BRAF V600E-mutant melanoma (A375) or colorectal cancer (COLO205) cell lines. Mice are randomized to receive vehicle or GNE-9815 at doses of 3, 10, or 30 mg/kg via oral gavage, once or twice daily, for 14-28 days. Tumor volume is measured twice weekly with calipers, and tumor growth inhibition is calculated. Body weight is monitored to assess tolerability. At study termination, tumors are harvested for analysis of p-ERK levels (by Western blot or IHC), Ki67 (proliferation marker), and cleaved caspase-3 (apoptosis marker). Blood and plasma are collected for pharmacokinetic analysis. Pharmacodynamic biomarkers are measured in plasma or tumor tissue to confirm target engagement and pathway modulation. |
| ADME/Pharmacokinetics |
Pharmacokinetic studies of GNE-9815 in rodents and non-human primates indicate that the compound is orally bioavailable with favorable half-life and exposure. Following oral administration in rodents at 10-30 mg/kg, the compound achieves peak plasma concentrations (Cmax) within 1-3 hours (Tmax) and has a plasma half-life of 4-8 hours. The oral bioavailability is approximately 50-80%, supporting once- or twice-daily dosing. The compound shows moderate plasma protein binding (approximately 70-90%) and distributes to tissues including tumor tissue. Metabolism is primarily via CYP450 enzymes (CYP3A4), and the compound is excreted in feces and urine. In preclinical studies, the compound demonstrates dose-proportional pharmacokinetics across the tested dose range.
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| Toxicity/Toxicokinetics |
Toxicology studies of GNE-9815 have been conducted in rodents and non-human primates as part of its preclinical development. At therapeutic doses (3-30 mg/kg/day) in repeat-dose studies, the compound is generally well-tolerated with no significant adverse effects on body weight, food consumption, hematology, or clinical chemistry parameters. At high doses (>50 mg/kg/day), mild to moderate increases in liver enzymes (ALT, AST) have been reported, along with gastrointestinal effects. No significant histopathological changes or organ toxicity have been observed at therapeutic doses. The compound does not appear to cause the cutaneous toxicity (skin rash) that is commonly associated with first-generation RAF inhibitors. The no-observed-adverse-effect level (NOAEL) is established at approximately 30-50 mg/kg/day in preclinical species, providing a safety margin for clinical development.
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| References | |
| Additional Infomation |
GNE-9815 (compound 7) is a next-generation pan-RAF inhibitor that represents a significant advancement in RAF-targeted therapy. The compound's type II binding mode and sub-nanomolar potency against both BRAF and CRAF address key limitations of first-generation RAF inhibitors, including paradoxical activation and acquired resistance. GNE-9815 has been investigated in preclinical and early clinical studies for the treatment of BRAF V600E-mutant and RAS-mutant cancers. The compound's favorable pharmacokinetic properties and safety profile support its continued development as a potential therapeutic option for patients with RAF- and RAS-driven malignancies. Ongoing research is focused on identifying the optimal patient populations and combination strategies for GNE-9815, including combinations with MEK inhibitors, immunotherapy, and other targeted agents.
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| Molecular Formula |
C26H22FN5O2
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|---|---|
| Exact Mass |
455.18
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| Elemental Analysis |
C, 68.56; H, 4.87; F, 4.17; N, 15.38; O, 7.02
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| CAS # |
2729996-45-4
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| Related CAS # |
2729996-45-4
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| PubChem CID |
155920128
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| Appearance |
Off-white to light yellow solid powder
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| LogP |
3.7
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
6
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
34
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| Complexity |
867
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC1=CC(=C(C=C1C2=CN=C3C(=C2)C=NN(C3=O)C)NC(=O)C4=CC(=CC=C4)C(C)(C)C#N)F
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| InChi Key |
SXCTZLXYGLXXRY-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C26H22FN5O2/c1-15-8-21(27)22(31-24(33)16-6-5-7-19(10-16)26(2,3)14-28)11-20(15)17-9-18-13-30-32(4)25(34)23(18)29-12-17/h5-13H,1-4H3,(H,31,33)
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| Chemical Name |
3-(2-cyanopropan-2-yl)-N-[2-fluoro-4-methyl-5-(7-methyl-8-oxopyrido[2,3-d]pyridazin-3-yl)phenyl]benzamide
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| Synonyms |
GNE 9815; GNE-9815; GNE9815
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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 (~109.8 mM)
Ethanol: ~5 mg/mL (11.0 mM) |
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.49 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 25.0 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: ≥ 2.5 mg/mL (5.49 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 25.0 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: ≥ 2.5 mg/mL (5.49 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
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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