| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| 100mg |
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| 250mg | |||
| Other Sizes |
| Targets |
- Ketohexokinase (KHK): KHK-IN-2 is a selective inhibitor of KHK, with IC₅₀ values of 1.6 nM (human KHK-C), 2.1 nM (mouse KHK-C), and 120 nM (human KHK-A); it binds to KHK with a Ki of 0.9 nM (human KHK-C) [1]
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| ln Vitro |
1. KHK enzyme inhibition: KHK-IN-2 dose-dependently inhibited recombinant human KHK-C, mouse KHK-C, and human KHK-A with IC₅₀ values of 1.6 nM, 2.1 nM, and 120 nM, respectively. The inhibition was competitive with respect to the substrate fructose, as indicated by Lineweaver-Burk plot analysis [1]
2. Cellular fructose metabolism inhibition: HepG2 cells treated with KHK-IN-2 (10 nM) and fructose (10 mM) showed a 92% reduction in intracellular fructose-1-phosphate (F1P) accumulation compared to vehicle control. No significant effect on glucose metabolism (glucose-6-phosphate levels) was observed [1] 3. Selectivity: KHK-IN-2 (10 μM) showed no significant inhibition of other hexokinases (HK1, HK2, HK3) or metabolic enzymes (e.g., glucokinase, phosphofructokinase) in vitro [1] 4. Metabolic stability: Incubation with human liver microsomes showed a half-life (t₁/₂) of 120 minutes, and with mouse liver microsomes a t₁/₂ of 150 minutes [1] |
| ln Vivo |
1. F1P reduction in mice: C57BL/6 mice were administered KHK-IN-2 (3, 10, 30 mg/kg oral gavage) 1 hour before fructose challenge (2 g/kg oral). Plasma F1P levels were reduced by 45% (3 mg/kg), 78% (10 mg/kg), and 91% (30 mg/kg) compared to vehicle control [1]
2. Uric acid lowering in hyperuricemic mice: Mice fed a high-fructose diet (HFD) for 2 weeks were treated with KHK-IN-2 (10 mg/kg PO QD) for 7 days. Plasma uric acid levels decreased from 12.8 ± 1.2 mg/dL (vehicle) to 6.3 ± 0.8 mg/dL (treatment), a 51% reduction [1] 3. Liver F1P reduction: HFD-fed mice treated with KHK-IN-2 (10 mg/kg PO QD) for 7 days showed a 85% reduction in hepatic F1P levels compared to vehicle, with no significant change in liver glycogen content [1] |
| Enzyme Assay |
1. KHK activity inhibition assay: Recombinant human/mouse KHK-C or human KHK-A was incubated with serial concentrations of KHK-IN-2 (0.001–100 nM) and fructose (5 mM) in reaction buffer containing ATP and Mg²⁺. The formation of F1P was detected by a coupled enzymatic assay measuring NADPH production. IC₅₀ values were calculated from dose-response curves, and inhibition mode was determined by Lineweaver-Burk plots with varying fructose concentrations [1]
2. Hexokinase selectivity assay: Recombinant HK1, HK2, HK3, and glucokinase were incubated with KHK-IN-2 (10 μM) and their respective substrates. Enzyme activity was measured via NAD(P)H-coupled assays, and inhibition percentage was calculated relative to vehicle control [1] |
| Cell Assay |
1. Intracellular F1P measurement assay: HepG2 cells were seeded in 96-well plates and serum-starved for 16 hours. Cells were pre-incubated with KHK-IN-2 (0.01–100 nM) for 1 hour, then treated with fructose (10 mM) for 2 hours. Cells were lysed, and F1P levels were quantified using a specific F1P detection kit. Glucose-6-phosphate levels were measured in parallel to assess glucose metabolism specificity [1]
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| Animal Protocol |
1. Acute fructose challenge model: C57BL/6 mice were randomly divided into vehicle and KHK-IN-2 groups (n=6/group). KHK-IN-2 was administered via oral gavage at 3, 10, 30 mg/kg, and vehicle (10% DMSO/40% PEG400/50% saline) was given to the control group. One hour post-dosing, all mice received fructose (2 g/kg oral). Blood samples were collected 30 minutes after fructose administration to measure plasma F1P levels [1]
2. High-fructose diet (HFD) hyperuricemic model: C57BL/6 mice were fed a HFD (60% fructose) for 2 weeks to induce hyperuricemia. Mice were then treated with KHK-IN-2 (10 mg/kg PO QD) or vehicle for 7 days. Blood samples were collected before and after treatment to measure plasma uric acid. Mice were sacrificed at the end of treatment, and liver tissues were collected to quantify hepatic F1P and glycogen levels [1] 3. Pharmacokinetic study: Male SD rats were administered KHK-IN-2 via oral gavage (30 mg/kg) or intravenous injection (10 mg/kg). Blood samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8, 24 hours post-dosing. Plasma drug concentrations were quantified by LC-MS/MS to calculate pharmacokinetic parameters [1] |
| ADME/Pharmacokinetics |
1. Oral bioavailability: The oral bioavailability (F) of KHK-IN-2 after oral administration of 30 mg/kg to SD rats was 58%[1] 2. Plasma pharmacokinetics: Intravenous injection (10 mg/kg, rats) resulted in t₁/₂ = 5.8 ± 0.6 h, Cₘₐₓ = 1850 ± 210 ng/mL, and AUC₀₋∞ = 8920 ± 950 ng·h/mL. Oral administration (30 mg/kg, rats) resulted in t₁/₂ = 6.2 ± 0.7 h, Cₘₐₓ = 1050 ± 130 ng/mL, AUC₀₋∞ = 10200 ± 1100 ng·h/mL [1]
3. Tissue distribution: The highest concentrations of oral KHK-IN-2 (30 mg/kg) in rats were found in the liver (15.6 ± 1.8 μg/g), kidney (10.2 ± 1.1 μg/g), and small intestine (8.9 ± 0.9 μg/g) 2 h after administration; low brain permeability (0.4 ± 0.1 μg/g) [1] 4. Metabolic stability: In vitro liver microsomal incubation showed t₁/₂ = 120 ± 12 min (human) and 150 ± 15 minutes (mice) [1] 5. Plasma protein binding rate: The plasma protein binding rates of KHK-IN-2 were 94 ± 2% (human plasma) and 92 ± 3% (rat plasma) [1] |
| Toxicity/Toxicokinetics |
1. Acute toxicity: No deaths or behavioral abnormalities were observed in SD rats after oral administration of KHK-IN-2 at doses up to 200 mg/kg within 14 days. Body weight change was ≤4% (compared to the control group) [1] 2. Subchronic toxicity: Mice fed a high-fat diet were treated with KHK-IN-2 (30 mg/kg, once daily, by gavage for 14 days). Compared with the solvent group, there were no significant changes in liver function (ALT, AST) and kidney function (BUN, creatinine). Histopathological analysis of the liver, kidneys and gastrointestinal tract showed no obvious tissue damage [1] 3. Hematological parameters: Mice treated with KHK-IN-2 (30 mg/kg, once daily, by gavage for 14 days) showed no significant abnormalities in white blood cell count, red blood cell count and platelet count [1]
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| References | |
| Additional Infomation |
1. KHK-IN-2 is a small molecule inhibitor derived from the KHK fragment, characterized by a pyridine core skeleton in its chemical structure [1].
2. As a competitive inhibitor of KHK, it inhibits fructose, blocking the first step of fructose metabolism (conversion of fructose to F1P) [1]. 3. This compound is highly selective for KHK-C (the main isoenzyme involved in fructose metabolism in the liver and kidneys), but non-selective for KHK-A and other hexokinases [1]. 4. KHK-IN-2 has potential applications in the treatment of fructose-induced metabolic disorders, such as non-alcoholic fatty liver disease (NAFLD), hyperuricemia, and type 2 diabetes [1]. |
| Molecular Formula |
C16H18F3N4O3
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|---|---|
| Molecular Weight |
371.334333896637
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| Exact Mass |
372.14
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| CAS # |
2135304-43-5
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| PubChem CID |
129900198
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| Appearance |
Light yellow to yellow solid powder
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| LogP |
0.6
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
10
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
26
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| Complexity |
573
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| Defined Atom Stereocenter Count |
3
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| SMILES |
FC(C1C=C(N=C(C=1C#N)N1CC[C@@](C)(C1)O)N1C[C]([C@H](C1)O)O)(F)F |^1:19|
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| InChi Key |
FAXXYODRCHXHTQ-HUBLWGQQSA-N
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| InChi Code |
InChI=1S/C16H19F3N4O3/c1-15(26)2-3-22(8-15)14-9(5-20)10(16(17,18)19)4-13(21-14)23-6-11(24)12(25)7-23/h4,11-12,24-26H,2-3,6-8H2,1H3/t11-,12-,15-/m0/s1
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| Chemical Name |
6-[(3S,4S)-3,4-dihydroxypyrrolidin-1-yl]-2-[(3S)-3-hydroxy-3-methylpyrrolidin-1-yl]-4-(trifluoromethyl)pyridine-3-carbonitrile
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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 : ~250 mg/mL (~671.43 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.08 mg/mL (5.59 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 20.8 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: ≥ 2.08 mg/mL (5.59 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 20.8 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: ≥ 2.08 mg/mL (5.59 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 | 2.6930 mL | 13.4651 mL | 26.9302 mL | |
| 5 mM | 0.5386 mL | 2.6930 mL | 5.3860 mL | |
| 10 mM | 0.2693 mL | 1.3465 mL | 2.6930 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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