| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
The primary target of Bestatin methyl ester is intracellular neutral aminopeptidase, where it acts as a competitive inhibitor by binding to the Zn²⁺ ion in the enzyme active site. Compared to Bestatin, the methyl ester modification results in significantly reduced inhibitory activity against basic aminopeptidase, indicating improved target selectivity for this derivative. Additionally, this compound promotes the degradation of cIAP1 protein.
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|---|---|
| ln Vitro |
Bestatin methyl ester (600, 900 µM; 24 h) inhibits spore cell differentiation in Dictyostelium discoideum [1].
Bestatin methyl ester exhibits biological activity in various in vitro models. In HT-1080 cells, it promotes cIAP1 degradation with an IC₅₀ value of 0.69 μM; in MV4-11 cells, the cytotoxicity IC₅₀ after 24-hour treatment is 3.9 μM. Regarding apoptosis regulation, Bestatin methyl ester potentiates the growth-inhibitory effects of the agonistic anti-Fas antibody CH11 more efficiently than Bestatin, suggesting that intracellular neutral aminopeptidase plays an important role in death ligand-induced apoptosis in solid tumor cells. Furthermore, at concentrations of 600-900 μM for 24 hours, this compound inhibits spore cell differentiation in Dictyostelium discoideum. |
| ln Vivo |
Based on studies of Bestatin, aminopeptidase inhibitors exhibit analgesic activity in animal models, with Bestatin showing an ED₅₀ of 240 μg following intracerebroventricular administration in mice. As a lipophilic derivative of Bestatin, Bestatin methyl ester theoretically possesses better tissue distribution properties, but specific data on its in vivo anti-tumor efficacy and pharmacodynamics require further investigation.
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| Enzyme Assay |
Enzyme activity is assessed using recombinant human neutral aminopeptidase and fluorogenic substrates such as Leu-AMC or Ala-AMC. Bestatin methyl ester is diluted to various concentrations in reaction buffer (e.g., 50 mM HEPES, pH 7.4, containing 100 μM ZnCl₂) and pre-incubated with the enzyme at 37°C for 10-15 minutes, followed by substrate addition to initiate the reaction. Fluorescence signals (excitation 360-380 nm, emission 440-460 nm) are continuously monitored using a fluorescence microplate reader. The inhibition rate is calculated, and IC₅₀ values are obtained by non-linear regression fitting.
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| Cell Assay |
Exponentially growing human tumor cells (e.g., HT-1080 fibrosarcoma cells or MV4-11 leukemia cells) are seeded into 96-well culture plates at appropriate densities. After overnight attachment, cells are treated with various concentrations of Bestatin methyl ester (e.g., 0.01-100 μM) for 24-72 hours. Cell viability is assessed using CellTiter-Glo luminescent assay or MTT assay with absorbance measured at 570 nm. For cIAP1 degradation assays, cells are harvested after 3 hours of treatment, and cIAP1 protein levels are detected by Western blot analysis.
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| Animal Protocol |
Referring to the in vivo protocol of Bestatin, 6-8-week-old female mice can be used to establish tumor-bearing models. When tumor volumes reach approximately 100 mm³, Bestatin methyl ester is administered orally or intraperitoneally. Dosing regimens can be based on Bestatin protocols, with once-daily administration for 2-3 consecutive weeks. Tumor volume and body weight are measured 2-3 times per week to calculate the tumor inhibition rate. At the end of the experiment, tumor tissues are collected for immunohistochemistry or Western blot analysis.
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| ADME/Pharmacokinetics |
As a lipophilic prodrug of Bestatin, Bestatin methyl ester exhibits improved lipophilicity and cellular membrane penetration through methyl ester modification. Regarding erythrocyte uptake, the methyl ester derivative can cross cell membranes more effectively due to its reduced polarity. Upon entry into cells, the methyl ester bond is likely hydrolyzed by intracellular esterases to generate the active form for inhibitory activity. Specific pharmacokinetic parameters such as plasma half-life, bioavailability, and metabolic pathways require further experimental determination.
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| Toxicity/Toxicokinetics |
Toxicological data on Bestatin methyl ester are currently limited. Based on the clinical safety profile of its parent compound Bestatin, Bestatin is generally well-tolerated at conventional doses. As a cell-permeable derivative, the toxicity profile of Bestatin methyl ester may be similar to that of Bestatin, but due to its improved cellular penetration, potential off-target effects warrant attention. In in vitro cytotoxicity assays, this compound exhibits an IC₅₀ of 3.9 μM at 24 hours in MV4-11 cells, indicating cytotoxicity at higher concentrations. This product is explicitly intended for research use only and is not for human therapeutic use.
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| References |
| Molecular Formula |
C17H26N2O4
|
|---|---|
| Molecular Weight |
322.40
|
| Exact Mass |
322.189
|
| CAS # |
65322-89-6
|
| PubChem CID |
9927170
|
| Appearance |
Off-white to light brown oitment
|
| LogP |
2.161
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| Hydrogen Bond Donor Count |
3
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
9
|
| Heavy Atom Count |
23
|
| Complexity |
381
|
| Defined Atom Stereocenter Count |
3
|
| SMILES |
CC(C)C[C@@H](C(=O)OC)NC(=O)[C@H]([C@@H](CC1=CC=CC=C1)N)O
|
| InChi Key |
WGNKTWBIPBOGLO-ILXRZTDVSA-N
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| InChi Code |
InChI=1S/C17H26N2O4/c1-11(2)9-14(17(22)23-3)19-16(21)15(20)13(18)10-12-7-5-4-6-8-12/h4-8,11,13-15,20H,9-10,18H2,1-3H3,(H,19,21)/t13-,14+,15+/m1/s1
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| Chemical Name |
methyl (2S)-2-[[(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]amino]-4-methylpentanoate
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| Synonyms |
Bestatin-amido-Me; 339186-54-8; (2S)-2-[[(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]amino]-N,4-dimethylpentanamide; (S)-2-((2S,3R)-3-Amino-2-hydroxy-4-phenylbutanamido)-N,4-dimethylpentanamide; orb1987800;
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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 | 3.1017 mL | 15.5087 mL | 31.0174 mL | |
| 5 mM | 0.6203 mL | 3.1017 mL | 6.2035 mL | |
| 10 mM | 0.3102 mL | 1.5509 mL | 3.1017 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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