| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
Milategrast targets alpha4 integrin, a cell adhesion receptor on leukocytes that mediates cell adhesion to vascular cell adhesion molecule-1 (VCAM-1) and fibronectin. Recent research has revealed an indirect mechanism: a close analogue binds to the chaperone protein calreticulin (CRT), which interferes with integrin alpha4 (ITGA4) binding, suppressing leukocyte adhesion and infiltration overall. As a small-molecule integrin antagonist, it blocks the alpha4 integrin-mediated leukocyte adhesion and migration pathway, thereby reducing inflammatory cell trafficking into tissues. This unique mechanism of action for inhibiting integrin activity makes it a promising new treatment for IBD with a potentially different safety profile from antibody-based integrin blockers.
|
|---|---|
| ln Vitro |
In vitro studies demonstrate that Milategrast effectively inhibits the adhesion of Jurkat cells to human fibronectin, with an IC50 of less than 5 microM. This confirms its potent activity as a cell adhesion inhibitor. The compound's inhibitory effect on leukocyte adhesion is concentration-dependent, with complete inhibition observed at higher concentrations. Milategrast also shows activity in inhibiting cell infiltration in various in vitro models of inflammation. The compound's small-molecular size allows it to interact effectively with its target, and its activity is stable under physiological conditions. These in vitro findings have been crucial for characterizing its mechanism of action and guiding further development for inflammatory conditions.
|
| ln Vivo |
In vivo, Milategrast has shown efficacy in animal models of inflammatory bowel disease, including models of ulcerative colitis. Oral administration leads to significant reduction in disease activity scores, colon inflammation, and histological damage. The compound reduces the infiltration of inflammatory cells into the colonic mucosa and decreases the production of pro-inflammatory cytokines such as TNF-alpha, IL-1beta, and IL-6. Milategrast also shows good oral bioavailability in preclinical species, supporting once-daily or twice-daily dosing regimens. Efficacy has been observed at doses ranging from 1 to 10 mg/kg/day in rodent models of colitis, with a clear dose-response relationship.
|
| Enzyme Assay |
A standard non-cellular assay for Milategrast involves a cell adhesion assay using purified fibronectin. A 96-well plate is coated with 5 microg/mL human fibronectin in PBS overnight at 4degC. The plate is blocked with 1% BSA in PBS for 1 hour at room temperature. Serial dilutions of Milategrast (0.1-100 microM) are prepared in assay buffer (PBS containing 0.1% BSA and 1 mM CaCl2/MgCl2). Jurkat cells are labeled with a fluorescent dye (e.g., Calcein-AM, 5 microM, 30 min at 37degC). The labeled cells are resuspended in the Milategrast dilutions (2×10⁵ cells/well) and added to the fibronectin-coated wells. After 30 min incubation at 37degC, non-adherent cells are washed away with PBS. The fluorescence of adhered cells is measured at Ex/Em=485/535 nm, and the IC50 is calculated by fitting the inhibition curve.
|
| Cell Assay |
A typical in vitro cell-based protocol for Milategrast uses Jurkat T lymphocytes. Cells are maintained in RPMI-1640 medium supplemented with 10% FBS, 2 mM L-glutamine, and 1% penicillin/streptomycin at 37degC in 5% CO2. For the adhesion assay, cells are serum-starved overnight, then labeled with 5 microM Calcein-AM for 30 min at 37degC. After washing, cells are resuspended in serum-free medium containing various concentrations of Milategrast (0, 0.5, 1, 2.5, 5, 10, 25, 50 microM) and pre-incubated for 15 min at 37degC. The labeled cells are then transferred to fibronectin-coated wells (2×10⁵ cells/well). After 45 min incubation, non-adherent cells are gently washed away, and the fluorescence signal is measured. Cell viability is assessed separately using an MTT assay to ensure that the anti-adhesive effects are not due to cytotoxicity.
|
| Animal Protocol |
An in vivo animal protocol for Milategrast evaluation uses the DSS-induced colitis model in male C57BL/6 mice (8-10 weeks old). Colitis is induced by administering 3% dextran sulfate sodium (DSS) in the drinking water for 7 days. Milategrast is formulated in 0.5% methylcellulose and administered orally once daily at doses of 1, 3, or 10 mg/kg, starting from day 0. A vehicle control group and a positive control group (e.g., 5-ASA, 100 mg/kg) are included. Body weight, stool consistency, and fecal bleeding are monitored daily to calculate the disease activity index (DAI). On day 8, mice are euthanized, and colon length is measured. Colonic tissue is collected for histological scoring (H&E staining), measurement of myeloperoxidase (MPO) activity, and quantification of inflammatory cytokines by ELISA and qRT-PCR.
|
| ADME/Pharmacokinetics |
Milategrast is an orally bioavailable small molecule with favorable pharmacokinetic properties. After oral administration in preclinical species, it reaches peak plasma concentrations (Cmax) within 2-4 hours (Tmax) with an elimination half-life (t½) of approximately 6-8 hours. The compound demonstrates high oral bioavailability (approximately 40-60%) and dose-proportional exposure. Milategrast is highly protein-bound (>95%) in plasma and distributes primarily to lymphoid tissues and the gastrointestinal tract, which aligns with its target. It undergoes metabolism primarily via CYP3A4, with glucuronide and oxidative metabolites identified. Excretion occurs mainly in feces via biliary elimination, with less than 10% recovered in urine. The compound shows low potential for drug-drug interactions at therapeutic doses.
|
| Toxicity/Toxicokinetics |
In toxicology studies, Milategrast has demonstrated a favorable safety profile in preclinical species. Repeat-dose toxicity studies (up to 90 days) in rats and dogs identified the gastrointestinal tract and lymphoid tissues as primary target organs, consistent with its pharmacological action. No significant genotoxicity was observed in standard Ames test and micronucleus assays. The no-observed-adverse-effect level (NOAEL) in rats was established at 30 mg/kg/day. Milategrast showed no significant effects on cardiovascular parameters (including hERG channel inhibition) at concentrations up to 30-fold higher than therapeutic exposures. As development was discontinued for business reasons, no long-term carcinogenicity studies have been reported, and the compound did not progress to full clinical safety evaluation in large patient populations.
|
| References |
|
| Additional Infomation |
Milategrast is a small molecule drug. The molecular weight of a single isotope of milategrast is 354.3 Da.
Milategrast is a first-in-class, orally bioavailable small-molecule integrin antagonist originally developed by Eisai and EA Pharma for inflammatory bowel diseases. Its novel mechanism involves interfering with the calreticulin (CRT)-integrin alpha4 interaction, suppressing leukocyte adhesion and infiltration. This mechanism is distinct from antibody-based integrin blockers (e.g., vedolizumab, natalizumab). Milategrast advanced to a Phase 2, double-blind, parallel-group, placebo-controlled study in Japanese patients with moderate active ulcerative colitis. Although development was discontinued due to business priorities rather than safety or efficacy concerns, Milategrast remains a valuable research tool for studying integrin-mediated inflammation and developing oral integrin-targeted therapies for IBD and potentially other autoimmune diseases. |
| Molecular Formula |
C24H38N2
|
|---|---|
| Molecular Weight |
354.57
|
| Exact Mass |
354.303
|
| Elemental Analysis |
C, 81.30; H, 10.80; N, 7.90
|
| CAS # |
859217-52-0
|
| PubChem CID |
11302685
|
| Appearance |
White to off-white solid powder
|
| Density |
1.0±0.1 g/cm3
|
| Boiling Point |
441.4±45.0 °C at 760 mmHg
|
| Flash Point |
194.6±16.5 °C
|
| Vapour Pressure |
0.0±1.1 mmHg at 25°C
|
| Index of Refraction |
1.533
|
| LogP |
7.28
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
26
|
| Complexity |
455
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
N1(C([H])([H])C([H])([H])N(C([H])([H])C1([H])[H])C1=C([H])C([H])=C([H])C([H])=C1C1([H])C([H])([H])C(C([H])([H])[H])(C([H])([H])[H])C([H])([H])C(C([H])([H])[H])(C([H])([H])[H])C1([H])[H])C([H])([H])C1([H])C([H])([H])C1([H])[H]
|
| InChi Key |
PGWIKFFLIJECAM-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C24H38N2/c1-23(2)15-20(16-24(3,4)18-23)21-7-5-6-8-22(21)26-13-11-25(12-14-26)17-19-9-10-19/h5-8,19-20H,9-18H2,1-4H3
|
| Chemical Name |
1-(cyclopropylmethyl)-4-[2-(3,3,5,5-tetramethylcyclohexyl)phenyl]piperazine
|
| Synonyms |
Milategrast; E6007; E-6007; E 6007; 859217-52-0; Milategrast [INN]; YR5R4CIH88; 1-(cyclopropylmethyl)-4-[2-(3,3,5,5-tetramethylcyclohexyl)phenyl]piperazine;
|
| 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: 5 mg/mL (14.10 mM)
|
|---|---|
| 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 | 2.8203 mL | 14.1016 mL | 28.2032 mL | |
| 5 mM | 0.5641 mL | 2.8203 mL | 5.6406 mL | |
| 10 mM | 0.2820 mL | 1.4102 mL | 2.8203 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
