| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| 500mg | |||
| Other Sizes |
Purity: =100%
| Targets |
MAP4K4 (IC50 = 3.7 nM)
GNE-495 is a strong MAP4K4 inhibitor that works well to promote angiogenesis. GNE-495 exhibits minimum body stability, permeability, efficient ideal cellular equilibrium, and MAP4K4 adsorption [1]. |
|---|---|
| ln Vitro |
GNE-495 is a strong MAP4K4 inhibitor that works well to promote angiogenesis. GNE-495 exhibits minimum body stability, permeability, efficient ideal cellular equilibrium, and MAP4K4 adsorption [1].
GNE-495 exhibited potent biochemical inhibition of MAP4K4 with an IC50 of 3.7 ± 1.4 nM. In a cellular assay using human umbilical vein endothelial cells (HUVEC), GNE-495 inhibited cell migration with an IC50 of 0.057 ± 0.004 µM, which is used to evaluate anti-angiogenic behavior. The compound maintained good kinase selectivity, consistent with its binding to a folded P-loop conformation of MAP4K4 that is observed in only a small subset of kinases. [1] |
| ln Vivo |
High doses of GNE-495—25 and 50 mg/kg—were given intraperitoneally to newborn mouse pups. In every species examined, GNE-495 demonstrates favorable internal characteristics, including low clearance, moderate terminal half-phase decay, and appropriate leverage levels (F=37–47%) [1].
Intraperitoneal (IP) injection of GNE-495 into newborn mice dose-dependently delayed retinal vascular outgrowth and induced abnormal retinal vascular morphology (e.g., long membrane protrusions in vascular endothelial cells). These in vivo phenotypes recapitulated the retinal vascular defects observed in inducible Map4k4 knockout mice, confirming the compound's in vivo activity and its on-target effect on MAP4K4. [1] |
| Enzyme Assay |
The biochemical IC50 against MAP4K4 was determined using a Z'-LYTE assay format, with data representing the average of at least three independent experiments.
For some compounds in the series, IC50 was also determined using Caliper LabChip 3000 (LC3K) technology. [1] |
| Cell Assay |
The anti-angiogenic cellular potency of GNE-495 was assessed using a HUVEC cell migration assay.
The assay measures the inhibition of cell migration, which is a key process in angiogenesis. The IC50 value reported is the concentration that inhibits 50% of cell migration. [1] |
| Animal Protocol |
Rats, Mice and Pups
For the brain cassette study, three male Sprague-Dawley (SD) rats receive an intravenous (IV) bolus dose of six test substances (for example, GNE-495; 0.5 mg/kg). GNE-495 is injected intravenously in bolus doses of 1 mg/kg in female CD-1 mice for the mouse PK study. GNE-495 (5 mg/kg) is additionally injected intravenously (PO) into female CD-1 mice. The rat brain cassette PK is dosed at 2 mL/kg, and all other doses are administered at 5 mL/kg. Water and food are available to animals at all times, and they are not fasted before receiving a dose. Three blood samples (~60 μL) are taken from each mouse up to either 9 or 24 hours after the compound of interest is given. This is done using a serial sampling method, and it involves taking three blood samples from each mouse. After being mixed with K2EDTA and placed on ice or in a cold Kryorack immediately after collection, the blood is then centrifuged to separate the plasma. Blood samples are centrifuged for 10–15 minutes at 4°C while being spun at a speed of 1000-2000× g within an hour of being drawn. This process yields the plasma. The plasma samples are stored at -70 to -80°C until analysis. For neonate PK, 3-day-old CD1 pups are injected with 25 mg/kg and 50 mg/kg GNE-495. Blood samples are taken intraperitoneally at the times specified, retinas are taken one hour after the dose, snap frozen in liquid nitrogen, and kept at -80°C until analysis. The concentrations of plasma and retinal lysate are assessed using LC/MS/MS. For the efficacy study in the neonatal retinal angiogenesis model, newborn mouse pups received intraperitoneal (IP) injections of GNE-495 daily from postnatal day 1 (P1) to P5 or P6. Doses used were 25, 50, and 100 mg/kg. The compound was formulated in a solution containing 5% DMSO and 95% corn oil. Retinas were harvested at P6 or P7, flat-mounted, and stained with Isolectin-B4 to visualize blood vessels for analysis of avascular area and vascular morphology. [1] |
| ADME/Pharmacokinetics |
In mice, after intravenous injection of 1 mg/kg GNE-495, its clearance (CL) was 19 mL/min/kg, terminal half-life (T1/2) was 1.5 h, volume of distribution (V) was 1.6 L/kg, and AUC extrapolated to infinity (AUCinf) was 5.3 hµM. After oral administration of 5 mg/kg in mice, the maximum plasma concentration (Cmax) was 1.4 µM, the time to peak concentration (Tmax) was 1.0 h, and the oral bioavailability (F%) was 47%. The extremely low free brain tissue concentration (0.008 µM) measured 1 h after administration indicates extremely low penetration into the central nervous system. In rats, the CL was 7.5 mL/min/kg, the T1/2 was 3.4 h, the V was 1.2 L/kg, and the oral bioavailability (F%) was 40%.
In dogs, the CL was 8.9 mL/min/kg, the T1/2 was 1.8 h, the V was 1.1 L/kg, and the oral F% was 37%. The compound had high plasma protein binding in mice (94.4%) and rats (97.8%). In vitro experiments showed that the compound had moderate permeability to MDCK cells (Papp = 7.3 x 10^-6 cm/s) and an efflux ratio of 2.3. The compound showed low metabolic clearance in both human and mouse liver microsomes (HLM CLint = 7 µL/min/mg; MLM CLint = 13 µL/min/mg). [1] |
| Toxicity/Toxicokinetics |
Predrug 1 was poorly tolerated in multi-day studies, presumably due to its significant central nervous system permeability (free brain concentration = 14.6 µM). GNE-495 was designed to minimize brain permeability (free brain concentration = 0.008 µM) to reduce this potential toxicity risk. The exact toxicity profile of GNE-495 in repeated-dose studies is not detailed in the literature. [1]
|
| References | |
| Additional Infomation |
GNE-495 (compound 13) is a structure-designed compound intended to reduce the penetration of MAP4K4 into the central nervous system (CNS) while maintaining its potent inhibitory effect by increasing the topological polar surface area (TPSA) and altering other physicochemical properties. X-ray eutectic structure (PDB: 42KS) confirmed its binding mode, indicating that the compound maintains a key interaction with the hinge region and induces a Tyr36-involved P-ring folding conformation. It is considered a valuable tool compound for studying the biological function of MAP4K4 in various diseases, including angiogenesis. [1]
|
| Molecular Formula |
C22H20FN5O2
|
|---|---|
| Molecular Weight |
405.4249
|
| Exact Mass |
405.16
|
| Elemental Analysis |
C, 65.18; H, 4.97; F, 4.69; N, 17.27; O, 7.89
|
| CAS # |
1449277-10-4
|
| Related CAS # |
1449277-10-4
|
| PubChem CID |
89730041
|
| Appearance |
Yellow solid powder
|
| Density |
1.5±0.1 g/cm3
|
| Boiling Point |
734.5±60.0 °C at 760 mmHg
|
| Flash Point |
398.0±32.9 °C
|
| Vapour Pressure |
0.0±2.4 mmHg at 25°C
|
| Index of Refraction |
1.712
|
| LogP |
2.54
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
30
|
| Complexity |
668
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
FC1=C([H])C([H])=C([H])C(=C1[H])C1C([H])=C([H])C2=C(C(N([H])[H])=NC([H])=C2C(N([H])C2([H])C([H])([H])N(C2([H])[H])C(C2([H])C([H])([H])C2([H])[H])=O)=O)N=1
|
| InChi Key |
FYXCIBJXJYBWPX-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C22H20FN5O2/c23-14-3-1-2-13(8-14)18-7-6-16-17(9-25-20(24)19(16)27-18)21(29)26-15-10-28(11-15)22(30)12-4-5-12/h1-3,6-9,12,15H,4-5,10-11H2,(H2,24,25)(H,26,29)
|
| Chemical Name |
8-amino-N-[1-(cyclopropanecarbonyl)azetidin-3-yl]-2-(3-fluorophenyl)-1,7-naphthyridine-5-carboxamide
|
| Synonyms |
GNE 495; GNE495; GNE-495
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| Solubility (In Vitro) |
DMSO: ~2.2 mg/mL (~5.4 mM)
|
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.22 mg/mL (0.54 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 2.2 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: ≥ 0.22 mg/mL (0.54 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 2.2 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: ≥ 0.22 mg/mL (0.54 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 | 2.4666 mL | 12.3329 mL | 24.6658 mL | |
| 5 mM | 0.4933 mL | 2.4666 mL | 4.9332 mL | |
| 10 mM | 0.2467 mL | 1.2333 mL | 2.4666 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.
 (A) Profile of pyridopyrimidine MAP4K4 inhibitor1;(B) X-ray structure of1in MAP4K4 (PDB: 4OBP).ACS Med Chem Lett.2015 Jun 29;6(8):913-8. |
|---|
 X-ray structure of13bound to MAP4K4 at 2.89 Å (PDB: 4ZK5).ACS Med Chem Lett.2015 Jun 29;6(8):913-8. |
 (A) Representative images of Isolectin-B4 (vascular marker) stained flat-mounted retinas at postnatal day 6 (P6) from mice treated with vehicle or 100 mg/kg GNE-495 daily from P1–P5.ACS Med Chem Lett.2015 Jun 29;6(8):913-8. |
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
COA