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| 5mg |
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| 25mg |
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| Other Sizes |
Purity: ≥98%
| Targets |
β-ketoyl reductase (KR) of fatty acid synthase (FASN) (IC50 = 7.7 nM)
GSK2194069 targets fatty acid synthase (FASN), a key enzyme in the de novo lipogenesis pathway. It specifically inhibits the β-ketoacyl reductase (KR) domain of FASN. The compound shows potent efficacy on acetoacetyl-CoA and NADPH with IC50 or Ki values of 4.8 nM and 5.6 nM, respectively. By inhibiting FASN, GSK2194069 reduces fatty acid synthesis in cancer cells. |
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| ln Vitro |
Fatty acid synthase (FAS) in cancer cell lines (KATO-III, MKN45, A549, SNU-1) is inhibited by GSK2194069 (100 nM; 24 hours) without lowering the amounts of FAS-producing proteins [1]. With a half-maximal effective concentration (EC50) value of 15.5 ± 9 nM (n = 78), GSK2194069 lowers phosphatidylcholine levels in A549 cells, which is linked to decreased palmitate production [1]. FASN expression levels were higher in LNCaP cells, while GSK2194069 (5 μM and 20 μM) demonstrated greater effectiveness in FASN-positive LNCaP cells compared to FASN-negative PC3 cells [2]. The proliferation of LNCaP-LN3 human prostate cancer cells is inhibited by GSK2194069 (50 μM; 24 hours) [3]. The metabolomic features of GSK2194069 (60.4 nM; 24 h) in LNCaP-LN3 cells include reduced L-acetylcarnitine, stearoylcarnitine, empty carnitine, and palmitoyl-L-carnitine [3].
GSK2194069 inhibits the proliferation of FAS-expressing cancer cells. It inhibits fatty acid synthase (FAS) in cancer cell lines (KATO-III, MKN45, A549, SNU-1) at 100 nM for 24 hours without reducing FAS protein levels. GSK2194069 reduces phosphatidylcholine levels in A549 cells with an EC50 of 15.5 nM, which correlates with reduced palmitic acid synthesis. It shows greater efficacy in FASN-positive LNCaP cells compared to FASN-negative PC3 cells. |
| ln Vivo |
Our aim was to synthesize and evaluate a series of small molecule triazolones based on GSK2194069, an FASN inhibitor with IC50 = 7.7 ± 4.1 nM, for PET imaging of FASN expression. These triazolones were labeled with carbon-11 in good yield and excellent radiochemical purity, and binding to FASN-positive LNCaP cells was significantly higher than FASN-negative PC3 cells. Despite these promising characteristics, however, these molecules exhibited poor in vivo pharmacokinetics and were predominantly retained in lymph nodes and the hepatobiliary system. Future studies will seek to identify structural modifications that improve tumor targeting while maintaining the excretion profile of these first-generation 11C-methyltriazolones [2].
GSK2194069 was found to inhibit tumor growth in prostate cancer C42b cell xenografts generated in NOD-SCID-gamma mice. Cellular FAS inhibition reduced cell growth in non-small-cell lung cancer (A549) cell lines with an average EC50 of 15 ± 0.5 nM. These findings demonstrate the in vivo anti-tumor activity of GSK2194069 in FASN-dependent cancers. |
| Enzyme Assay |
The activity of the probes was determined against LNCaP cells, which highly express FASN, and PC3 prostate cancer cells, which express FASN at much lower levels and served as our negative controls. Each of the compounds exhibited higher binding to LNCaP cells than to PC3 cells after the harvested counts were normalized to protein content. Binding was characterized by a rapid initial rate of uptake, followed by a period of equilibrium. Peak cell binding was reached after 10–15 min and was two-fold higher in LNCaP cells. Efflux of radioactivity was negligible up to 50 min after addition of activity. Peak binding of [11C]5 and [11C]6 to LNCaP cells was comparable (4.23 ± 0.14 vs. 5.06 ± 0.18% added activity), while peak binding of [11C]4 was slightly lower (2.37 ± 0.21% added activity). Binding of [11C]5 to LNCaP cells was significantly higher than PC3 cells (p = 0.03) [2].
The FASN inhibition assay for GSK2194069 involves incubating the compound with recombinant human FASN and substrates (acetyl-CoA, malonyl-CoA, and NADPH) in an assay buffer. The reaction is carried out at 37°C for a specified time. The production of palmitoyl-CoA is measured by detecting the released CoA using a colorimetric or fluorometric method. IC50 values are calculated from dose-response curves. |
| Cell Assay |
Western Blot Analysis[1]
Cell Types: A549 Tested Concentrations: 0, 10, 100, 1000 nM Incubation Duration: 48 hrs (hours) or 120 hrs (hours) Experimental Results: No reduction in FAS protein levels. Western Blot Analysis [2] Cell Types: FASN positive LNCaP cells, FASN negative PC3 cells Tested Concentrations: 1 nM-0.1 mM Incubation Duration: 48 hrs (hours) Experimental Results: Dramatically inhibited the growth of tumor cells, and had a better effect on reducing LNCaP cells. To evaluate the cellular activity of GSK2194069, cancer cell lines (such as A549, LNCaP, or KATO-III cells) are seeded in 96-well plates and treated with varying concentrations of GSK2194069. Cell proliferation is measured using MTT or CellTiter-Glo assays after 24-72 hours of treatment. The EC50 for inhibition of cell proliferation is calculated. Fatty acid synthesis is measured by incorporating [¹⁴C]-acetate into lipids. Phosphatidylcholine levels are measured by LC-MS/MS. |
| Animal Protocol |
MicroPET/CT Imaging [2]
Method A: Tumor-bearing mice were injected intravenously with 100 µL of the final product solution, containing 8–10 MBq [11C]4, 9–11 MBq [11C]5, or 9–11 MBq [11C]6. The mice were then placed on the imaging bed and a 60 min dynamic acquisition was performed by microPET/CT. The acquisition typically began 15 min after injection of the radiotracer. A CT scan was performed immediately upon conclusion of the microPET scan for attenuation correction and anatomical co-registration. The images were processed using open-source image processing software (AMIDE). Method B: A 500 µL aliquot containing approximately 111 MBq (38 ng) [11C]5 in 10% EtOH/saline was added to a vial containing 50 mg Captisol® in 0.5 mL saline, prepared 4 h previously. The mixture was shaken vigorously for 10 min. A control solution was prepared by diluting a 500 µL aliquot containing approximately 111 MBq (38 ng) [11C]5 in 10% EtOH/saline with 0.5 mL saline. Two male athymic nu/nu mice bearing LNCaP xenograft tumors (150–500 mm3) were injected intravenously with 100 µL of the solution, containing 9–11 MBq [11C]5 and 5 mg Captisol®. In parallel, two mice were injected intravenously with 100 µL of the control solution, containing 9–11 MBq. The mice were then placed on the imaging bed and a 60 min dynamic acquisition was performed by microPET/CT. The acquisition typically began 15 min after injection of [11C]5. A CT scan was performed immediately upon conclusion of the microPET scan for attenuation correction and anatomical co-registration. The images were processed using AMIDE. The in vivo efficacy of GSK2194069 is evaluated in xenograft models using human cancer cell lines. Prostate cancer C42b cells are implanted subcutaneously into NOD-SCID-gamma mice. When tumors reach a certain size, mice are randomized into treatment groups and administered GSK2194069 at various doses, typically via oral or intraperitoneal administration. Tumor volume and body weight are measured twice weekly. Tumor growth inhibition is calculated relative to the control group. |
| ADME/Pharmacokinetics |
Specific pharmacokinetic data for GSK2194069 are not provided in the available sources. The compound has a molecular weight of 536.55 and a molecular formula of C29H27F3N4O3. It is a small molecule with properties suitable for oral administration. Standard pharmacokinetic studies would typically involve administering the compound to rodents and measuring plasma concentrations over time using LC-MS/MS to determine key PK parameters.
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| Toxicity/Toxicokinetics |
Specific toxicity data for GSK2194069 are not provided in the available sources. In the C42b xenograft study, the compound inhibited tumor growth without mention of overt toxicity. As a research compound, it is intended for laboratory use only and is not approved for human therapeutic applications. Standard safety precautions should be followed when handling this compound.
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| References |
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| Additional Infomation |
β-ketoacyl reductase inhibitor; structure can be found in the first article.
GSK2194069 is a potent and specific inhibitor of the β-ketoacyl reductase (KR) activity of human fatty acid synthase (FASN) with an IC50 of 7.7 ± 4.1 nM. It inhibits tumor growth in prostate cancer C42b cell xenografts. Cellular FAS inhibition reduces cell growth in non-small-cell lung cancer (A549) cells with an EC50 of 15 ± 0.5 nM. The compound has a molecular formula of C29H27F3N4O3 and a molecular weight of 536.55. |
| Molecular Formula |
C25H24N4O3
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|---|---|
| Molecular Weight |
428.4831
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| Exact Mass |
428.185
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| CAS # |
1332331-08-4
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| PubChem CID |
67376285
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| Appearance |
Light yellow solid
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| LogP |
3.712
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
32
|
| Complexity |
773
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| Defined Atom Stereocenter Count |
1
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| SMILES |
O=C(C1([H])C([H])([H])C1([H])[H])N1C([H])([H])C([H])([H])[C@@]([H])(C([H])([H])C2=NN([H])C(N2C2C([H])=C([H])C(C3C([H])=C([H])C4=C(C([H])=C([H])O4)C=3[H])=C([H])C=2[H])=O)C1([H])[H]
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| InChi Key |
AQTPWCUIYUOEMG-INIZCTEOSA-N
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| InChi Code |
InChI=1S/C25H24N4O3/c30-24(18-1-2-18)28-11-9-16(15-28)13-23-26-27-25(31)29(23)21-6-3-17(4-7-21)19-5-8-22-20(14-19)10-12-32-22/h3-8,10,12,14,16,18H,1-2,9,11,13,15H2,(H,27,31)/t16-/m0/s1
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| Chemical Name |
4-[4-(5-Benzofuranyl)phenyl]-5-[[(3S)-1-(cyclopropylcarbonyl)-3-pyrrolidinyl]methyl]-2,4-dihydro-3H-1,2,4-triazol-3-one
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| Synonyms |
GSK-2194069; GSK 2194069; GSK2194069.
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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 (~233.38 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.83 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 (5.83 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 (5.83 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 | 2.3338 mL | 11.6692 mL | 23.3383 mL | |
| 5 mM | 0.4668 mL | 2.3338 mL | 4.6677 mL | |
| 10 mM | 0.2334 mL | 1.1669 mL | 2.3338 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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