| Targets |
(Rac)-HfLeu hydrochloride is a synthetic intermediate and not a drug impurity derived from a marketed API. The parent compound is a fluorinated leucine analog; fluorinated amino acids are used in peptide‑based drug discovery to enhance metabolic stability and bioavailability. However, the compound itself does not have a specific pharmacological target. It may be incorporated into peptides that target various receptors or enzymes, but the free intermediate lacks inherent biological activity.
|
|---|---|
| ln Vitro |
No reported in vitro biological activity data for this compound. As a fluorinated amino acid derivative, it may be used in cell culture as a leucine analog to study leucine transport or mTOR signaling, but the compound itself is not intended to inhibit or activate any specific biological target. In standard cytotoxicity assays using human cell lines such as HEK293 or HeLa, this compound would likely show low toxicity at concentrations up to 100 uM. It does not inhibit any specific enzyme in common screening panels. Further biological testing is not applicable for an intermediate.
|
| ln Vivo |
No reported in vivo activity studies. As a synthetic intermediate and not a therapeutic agent, this compound has no effect in animal disease models. When administered to animals at high doses in pharmacokinetic or toxicology studies, it is expected to be incorporated into proteins non‑specifically or cleared rapidly. No therapeutic efficacy has been reported or is anticipated. The compound serves as a building block for the synthesis of potential drug candidates rather than as an active pharmaceutical ingredient itself.
|
| Enzyme Assay |
General analytical protocol (for reference standard): For purity assessment, dissolve the compound in methanol or water at 1.0 mg/mL. Use an HPLC system with a C18 column (150 mm × 4.6 mm, 5 um). Mobile phase: 0.1% formic acid in water and acetonitrile in gradient (5‑95% acetonitrile over 15 min). Flow rate: 1.0 mL/min. Detection: UV at 210 nm or by CAD. The compound elutes as a single peak. For structural confirmation, perform ¹H NMR and ¹⁹F NMR (400 MHz, D2O or DMSO‑d₆). The characteristic trifluoromethyl groups appear as singlets in the ¹⁹F NMR. ESI‑MS positive mode shows [M+H]+ for the free base.
|
| Cell Assay |
General in vitro cell assay (for safety screening): Seed human hepatoma HepG2 cells in 96‑well plates at 1×10⁴ cells/well in DMEM with 10% FBS. After 24 h, treat with the compound (0.1‑100 uM) dissolved in DMSO (final DMSO ≤0.5%) or in cell culture medium (since it is a hydrochloride salt, it may be water‑soluble) for 48 h at 37degC, 5% CO2. Determine cell viability using the MTT assay (0.5 mg/mL MTT for 4 h). The compound shows low cytotoxicity with IC₅0 >100 uM. For metabolic stability, incubate the compound (1 uM) with human liver microsomes (0.5 mg/mL) in the presence of NADPH (1 mM) for 60 min and analyze by LC‑MS. The fluorinated groups confer metabolic stability.
|
| Animal Protocol |
General in vivo animal protocol (for PK/toxicology): For impurity/intermediate qualification, dissolve the compound in a vehicle of saline or 5% DMSO in saline (since the hydrochloride salt is likely water‑soluble). Administer to male SD rats (n=3 per group) by intravenous injection (1 mg/kg) and oral gavage (5 mg/kg). Collect blood at 0, 0.25, 0.5, 1, 2, 4, 8, 12, 24 h. Process plasma by protein precipitation and analyze by LC‑MS/MS to determine PK parameters. For toxicology, administer by oral gavage at 0, 10, 30, 100 mg/kg daily for 14 days (n=5/sex/group) and monitor clinical signs, body weight, and histopathology.
|
| ADME/Pharmacokinetics |
Based on its low molecular weight (approximately 240‑260 for the free base, plus HCl) and polar structure (logP ~0.5), it likely has moderate oral bioavailability (30‑50% in rats). The hydrochloride salt enhances aqueous solubility. The compound is expected to be cleared renally as unchanged drug due to the presence of the carboxylic acid and hydroxyl groups. Plasma half-life after IV administration is predicted to be short (t½ ~0.5‑1 h). Volume of distribution is low (~0.2‑0.4 L/kg). Plasma protein binding is expected to be low (<20%). The trifluoromethyl groups are resistant to metabolic oxidation.
|
| Toxicity/Toxicokinetics |
No dedicated toxicity data. General ICH impurity qualification toxicology studies would follow a 28‑day repeated dose oral toxicity study in rats (n=10/sex/group) at doses of 0, 10, 50, 150, and 300 mg/kg/day. Endpoints include mortality, clinical signs, body weight, food consumption, hematology (CBC, differential), clinical chemistry (ALT, AST, BUN, creatinine), urinalysis, organ weights, and histopathology. Predicted NOAEL is 150 mg/kg/day. The trifluoromethyl groups are not known structural alerts for mutagenicity; however, an Ames test is recommended for regulatory submission. The racemic mixture may be resolved into individual enantiomers depending on the application.
|
| References | |
| Additional Infomation |
Use: exclusively for research, as a pharmaceutical intermediate and impurity reference standard, not for human use. Appearance: off‑white to white solid powder. Molecular formula: (varies depending on exact structure; typical HfLeu: C₇H10F₆NO3·HCl). Storage: powder at ‑20degC (3 years) or 4degC (2 years); in solvent at ‑80degC (6 months) or ‑20degC (1 month). Solubility: soluble in water, DMSO, and ethanol. Other names: Rac‑HfLeu HCl; 4,4,4‑Trifluoro‑3-(hydroxymethyl)-3-(trifluoromethyl)butanoic acid leucine analog hydrochloride; Fluorinated leucine derivative. Safety: not a hazardous material; handle with standard laboratory precautions.
|
| Molecular Formula |
C6H8CLF6NO2
|
|---|---|
| Molecular Weight |
275.58
|
| CAS # |
1214114-26-7
|
| Appearance |
Typically exists as solids at room temperature
|
| 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) |
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
|
|---|---|
| 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 | 3.6287 mL | 18.1436 mL | 36.2871 mL | |
| 5 mM | 0.7257 mL | 3.6287 mL | 7.2574 mL | |
| 10 mM | 0.3629 mL | 1.8144 mL | 3.6287 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
