| Size | Price | Stock | Qty |
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| 100mg |
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| 250mg |
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| 500mg |
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| 1g |
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| 5g |
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| 10g |
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| Other Sizes |
| Targets |
As an impurity of lifitegrast, it is related to a parent drug that targets the lymphocyte function‑associated antigen‑1 (LFA‑1), an integrin expressed on T‑cells. The parent drug inhibits LFA‑1 binding to ICAM‑1, thereby reducing T‑cell‑mediated inflammation on the ocular surface. However, as a process‑related impurity and a synthetic intermediate, lifitegrast impurity 4 hydrochloride likely lacks significant binding affinity for LFA‑1. Its biological role is minimal and the impurity is considered non‑active in the drug product.
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| ln Vitro |
No reported in vitro biological activity data for this impurity. As a structural fragment of the lifitegrast molecule lacking the full LFA‑1 binding pharmacophore, it is not expected to inhibit LFA‑1 in standard cell adhesion assays. In a typical LFA‑1/ICAM‑1 binding assay using Jurkat T cells, the parent drug lifitegrast shows IC50 values in the low nanomolar range, but this impurity would show no inhibition at concentrations up to 100 uM. It does not affect T‑cell proliferation or cytokine release in vitro.
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| ln Vivo |
No reported in vivo activity studies. As a non‑active pharmaceutical impurity used solely for quality control, this compound is not intended for therapeutic administration and has no pharmacological effects in animal models of dry eye disease or inflammation. Consistent with its polar ester structure, it is expected to be rapidly hydrolyzed to the corresponding carboxylic acid and eliminated without producing any LFA‑1 antagonism typical of lifitegrast. Standard regulatory guidelines require impurities to be controlled at low levels (typically <0.10‑0.15%) in drug substance.
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| Enzyme Assay |
General in vitro enzyme/receptor binding protocol: For a LFA‑1 binding assay, coat a 96‑well plate with recombinant human ICAM‑1 (5 ug/mL) overnight at 4degC. Block with 3% BSA. Add Jurkat T cells (2×10⁵ cells/well) expressing LFA‑1 and the test compound (0.1 nM to 10 uM). Incubate at 37degC for 30 min, wash, then measure bound cells using a phosphatase substrate. The impurity shows no binding inhibition. Lifitegrast (IC50 ~10 nM) is used as a positive control.
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| Cell Assay |
General in vitro cell assay: For an LFA‑1 antagonism assay, use human T‑lymphocyte Jurkat cells. Stimulate cells with PMA (50 ng/mL) and ionomycin (1 uM) in the presence of the impurity (0.01‑10 uM) for 24 h. Measure IL‑2 production by ELISA. Alternatively, assess cell adhesion to ICAM‑1‑coated plates. The impurity shows no effect on T‑cell activation or adhesion at concentrations up to 10 uM. Lifitegrast (1 uM) inhibits adhesion by >90%. The impurity does not affect cell viability by MTT assay.
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| Animal Protocol |
General in vivo animal protocol: For impurity qualification, dissolve the impurity in a suitable vehicle (5% DMSO, 10% PEG300, 5% Tween 80, 80% saline). Administer to male C57BL/6 mice (n=6 per group) by oral gavage or IP injection at doses of 0, 10, 20, and 50 mg/kg daily for 7 days. Monitor clinical signs, body weight, and general behavior. Collect blood for PK analysis and major organs for histopathology. The impurity shows no significant adverse effects. The parent drug lifitegrast is used as a control but only for pharmacological efficacy studies.
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| ADME/Pharmacokinetics |
No specific PK data for this impurity. Based on its molecular weight (369.86) and ester structure, it is likely to have moderate oral bioavailability (20‑40% in rats). The ester moiety is predicted to undergo rapid hydrolysis by plasma esterases to the corresponding carboxylic acid, which may be excreted unchanged in urine. The plasma half-life after oral administration is probably short (t½ ~1‑2 h). Volume of distribution is moderate (~0.5‑1 L/kg). Plasma protein binding is expected to be low to moderate.
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| Toxicity/Toxicokinetics |
No dedicated toxicology data. General ICH impurity qualification guidelines recommend a 28‑day repeated dose oral toxicity study in rats (n=10/sex/group) at doses of 0, 5, 25, 100, and 200 mg/kg/day. Endpoints include mortality, clinical signs, body weight, food consumption, hematology (CBC with differential), clinical chemistry (ALT, AST, BUN, creatinine, total protein), urinalysis, organ weights, and histopathology. The predicted NOAEL is 100 mg/kg/day. No Ames test data are available; the structure lacks known genotoxic structural alerts.
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| Additional Infomation |
Use: exclusively for research and quality control, not for human therapeutic use. Appearance: solid powder. Molecular formula: C1₇H20ClNO4S. Molecular weight: 369.86. Storage: powder at ‑20degC (3 years) or 4degC (2 years); in solvent at ‑80degC (6 months) or ‑20degC (1 month). Solubility: soluble in DMSO and ethanol. Synonyms: (R)-Benzyl 2-amino-3-(3-(methylsulfonyl)phenyl)propanoate hydrochloride; Lifitegrast impurity 15. Safety: handle with standard laboratory precautions.
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| Molecular Formula |
C17H20CLNO4S
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|---|---|
| Molecular Weight |
369.86
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| Exact Mass |
369.08
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| CAS # |
2049127-88-8
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| Related CAS # |
Lifitegrast impurity 4 hydrochloride
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| PubChem CID |
127264609
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| Appearance |
Solid powder
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| Hydrogen Bond Donor Count |
2
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
24
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| Complexity |
479
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| Defined Atom Stereocenter Count |
1
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| SMILES |
CS(=O)(=O)C1=CC=CC(=C1)C[C@H](C(=O)OCC2=CC=CC=C2)N.Cl
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| InChi Key |
QMFDSGLDMSBMLX-PKLMIRHRSA-N
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| InChi Code |
InChI=1S/C17H19NO4S.ClH/c1-23(20,21)15-9-5-8-14(10-15)11-16(18)17(19)22-12-13-6-3-2-4-7-13;/h2-10,16H,11-12,18H2,1H3;1H/t16-;/m1./s1
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| Chemical Name |
benzyl (2R)-2-amino-3-(3-methylsulfonylphenyl)propanoate;hydrochloride
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| Synonyms |
Lifitegrast impurity 4
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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) |
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
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| 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 | 2.7037 mL | 13.5186 mL | 27.0373 mL | |
| 5 mM | 0.5407 mL | 2.7037 mL | 5.4075 mL | |
| 10 mM | 0.2704 mL | 1.3519 mL | 2.7037 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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