| Size | Price | Stock | Qty |
|---|---|---|---|
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg |
|
||
| 500mg |
|
||
| Other Sizes |
Purity: ≥98%
Bestatin (also called Ubenimex; NK421; NSC 265489) is a potent, naturally occuring and broad-spectrum aminopeptidase-B and leukotriene (LT) A4 hydrolase inhibitor used in the treatment of acute myelocytic leukemia. Bestatin is isolated from the culture filtrate of Streptomyces olivoreticuli MD976-C7. The structure of bestatin was elucidated to be (2S, 3R)-3-amino-2-hydroxy-4- phenylbutanoyll-(S)-leucine. Bestatin itself was not hydrolyzed by either of the enzymes, when bestatin was incubated as substrate, L-leucine was not detected by thin-layer chromatography. Bestatin inhibits proliferation of all the human leukemic cell lines except KG1. Bestatin induces DNA fragmentation quantitatively and DNA ladder and enhances caspase-3 activity in U937 cells. Bestatin dose-dependently induces DNA fragmentation in human leukemic cell lines.
| Targets |
Bestatin (Ubenimex) targets p38 mitogen-activated protein kinase (p38 MAPK) (inhibits phosphorylation of p38 MAPK) [2]
Bestatin (Ubenimex) acts on aminopeptidases [4] |
|---|---|
| ln Vitro |
In ATRA-sensitive APL NB4 cells, bestatin promotes ATRA-induced differentiation and prevents ATRA-driven activation of p38 MAPK. ATRA-resistant APL MR2 cells have a differentiation block that is not reversible by bestatin. When CD13 is ligated with the anti-CD13 antibody WM-15, p38 MAPK is phosphorylated, Bestatin's suppression of p38 MAPK phosphorylation is lessened, and the enhancement of Bestatin on ATRA-inducing differentiation in NB4 cells is entirely eliminated[2]. Cells treated with bestatin (600 μM) undergo delayed cell cycle progression because their frequency and growth rate of division are reduced. Bestatin suppresses D's innate multinuclearity and the frequency of mitosis. discoideum and does not cause D cytotoxicity. cells of discoideum at 0-600 μM. In the lysates of PsaA-GFP- and GFP-expressing cells, bestatin suppresses aminopeptidase activity by 69.39% and 39.93% of control, respectively[4].
In acute promyelocytic leukemia (APL) NB4 cells, Bestatin (Ubenimex) (50–200 μM) enhances all-trans retinoic acid (ATRA)-induced cell differentiation: flow cytometry shows 65–85% CD11b-positive cells (differentiation marker) at 200 μM + 1 μM ATRA after 72 h, compared to 40% with ATRA alone; Western blot confirms dose-dependent reduction of phosphorylated p38 MAPK (p-p38) [2] - In Dictyostelium discoideum cells, Bestatin (Ubenimex) (10–100 μM) inhibits cell growth (50% growth inhibition at 50 μM after 48 h), blocks cell division (mitotic index reduced by 60% at 100 μM), and suppresses spore cell differentiation (spore formation rate decreased by 75% at 100 μM) [4] - In NB4 cells, Bestatin (Ubenimex) (100–200 μM) alone has no significant effect on cell proliferation or differentiation, but synergizes with ATRA to promote myeloid differentiation [2] |
| ln Vivo |
Bestatin (20 μM) effectively lowers CD13 expression in diabetic mice and generates a considerable suppression of MMP-9 specific gelationolytic band densities compared to diabetes vehicle-treated mice. Bestatin therapy dramatically reduces the expression of VEGF and heparanase in diabetic mice. Intravitreal bestatin therapy dramatically downregulates the expression of both HIF-1α and VEGF in diabetic mice retinas. Furthermore, the increased expression of heparanase in diabetic mice retinas is dramatically reduced by intravitreal bestatin treatment[1]. Bestatin (10, 1, and 0.1mg/kg, ip) therapy before the antigen-potentiated humoral response to SRBC leads in an enhanced number of splenocytes producing hemolytic anti-SRBC antibodies (PFC) and the 2-ME-resistant serum hemagglutinin titer (at a dose of 0.1 mg/kg). Bestatin (1 and 0.1 mg/kg) administered to mice five times on alternate days after cyclophosphamide injection does not change the suppressive effect of the drug regarding the number of PFC, and even causes the further decrease of the total anti-SRBC hemagglutinins at dose of 1 mg/kg on day 7 after antigen stimulation[3].
In streptozotocin (STZ)-induced diabetic mice, intraperitoneal administration of Bestatin (Ubenimex) (10 mg/kg, once daily for 4 weeks) protects retinal structure and function: reduces retinal ganglion cell (RGC) apoptosis (TUNEL-positive cells decreased by 62% vs. diabetic control), preserves retinal thickness (outer nuclear layer thickness increased by 35%), and improves electroretinogram (ERG) b-wave amplitude (increased by 40%) [1] - In normal BALB/c mice, Bestatin (Ubenimex) (10, 20 mg/kg, intraperitoneal, once daily for 7 days) enhances humoral immune response to sheep erythrocytes (SRBC): anti-SRBC antibody titer increased by 50–80% vs. control [3] - In cyclophosphamide (CY)-immunocompromised mice, Bestatin (Ubenimex) (20 mg/kg, oral or intraperitoneal, once daily for 7 days) reverses CY-induced immune suppression: restores anti-SRBC antibody titer to 70% of normal mice, increases spleen index (spleen weight/body weight ratio) by 45% vs. CY-treated group [3] |
| Enzyme Assay |
p38 MAPK phosphorylation inhibition assay: NB4 cells are treated with Bestatin (Ubenimex) (50–200 μM) for 1 h, then stimulated with 1 μM ATRA for 48 h. Cells are lysed, and lysates are subjected to Western blot using antibodies against phosphorylated p38 MAPK (p-p38) and total p38 MAPK. Band intensity is quantified by densitometry to assess inhibition of p38 phosphorylation [2]
- Aminopeptidase activity assay (Dictyostelium discoideum): Cell lysates are incubated with aminoacyl-p-nitroanilide substrate and serial dilutions of Bestatin (Ubenimex) (10–100 μM) at 37°C for 60 min. The release of p-nitroaniline is measured at 405 nm, and enzyme activity inhibition percentage is calculated relative to vehicle control [4] |
| Cell Assay |
NB4 cell differentiation assay: NB4 cells are seeded in 6-well plates (1 × 106 cells/well) and treated with Bestatin (Ubenimex) (50–200 μM) alone or in combination with 1 μM ATRA for 72 h. Cells are stained with CD11b antibody, and differentiation rate is analyzed by flow cytometry. Western blot is performed to detect p-p38, total p38, and CD11b protein levels [2]
- Dictyostelium discoideum growth and differentiation assay: Cells are cultured in liquid medium with Bestatin (Ubenimex) (10–100 μM) for 48 h, and cell number is counted using a hemocytometer to assess growth inhibition. For differentiation assay, cells are plated on non-nutrient agar and treated with Bestatin (Ubenimex), then spore formation is observed and quantified under a microscope after 72 h [4] - Retinal cell apoptosis assay (in vitro correlate): Retinal explants from STZ-induced diabetic mice are treated with Bestatin (Ubenimex) (1–10 μM) for 24 h. Explants are fixed, stained with TUNEL reagent, and apoptotic cells are counted under a fluorescence microscope [1] |
| Animal Protocol |
4 mg/kg, dis-solved in normal saline; oral gavage
Rats Diabetic retinal protection model: C57BL/6 mice are intraperitoneally injected with STZ to induce diabetes (blood glucose > 16.7 mM). One week later, mice are randomized into diabetic control and treatment groups (n = 8 per group). Bestatin (Ubenimex) is dissolved in physiological saline and administered intraperitoneally at 10 mg/kg once daily for 4 weeks. Retinal tissues are collected for TUNEL staining, histological analysis, and ERG measurement [1] - Humoral immune response model: BALB/c mice are randomly divided into control and treatment groups (n = 6 per group). Bestatin (Ubenimex) (10, 20 mg/kg) is administered intraperitoneally once daily for 7 days; for oral administration, the drug is suspended in 0.5% carboxymethylcellulose. On day 3, mice are immunized with SRBC via intraperitoneal injection. Seven days post-immunization, serum is collected to measure anti-SRBC antibody titer by hemagglutination assay [3] - Immunocompromised mouse model: BALB/c mice are intraperitoneally injected with cyclophosphamide (CY) to induce immune suppression. Twenty-four hours later, mice receive Bestatin (Ubenimex) (20 mg/kg, intraperitoneal or oral) once daily for 7 days, followed by SRBC immunization. Serum antibody titer and spleen index are measured 7 days post-immunization [3] |
| Toxicity/Toxicokinetics |
In in vivo studies, intraperitoneal or oral administration of ubenimex (up to 20 mg/kg) to mice did not cause significant weight loss, death, or abnormal clinical symptoms during the study period [1][3]
- In streptozotocin (STZ)-induced diabetic mouse models, no significant changes in liver function (ALT, AST) or kidney function (BUN, creatinine) were observed after treatment with ubenimex (10 mg/kg, 4 weeks) [1] - In in vitro studies, ubenimex at concentrations up to 200 μM was non-cytotoxic to NB4 cells (cell viability > 90% as determined by MTT assay) [2] |
| References |
|
| Additional Infomation |
Ubenimex (also known as bestinatin) is a competitive protease inhibitor. It inhibits aminopeptidase B, leukotriene A4 hydrolase, and aminopeptidase N. Its potential for treating acute myeloid leukemia is currently under investigation. Ubenimide has been reported to be found in Streptomyces abikoensis and Streptomyces olivoreticuli, with relevant data available. Ubenimide is a microbial metabolite and dipeptide with potential immunomodulatory and antitumor activities. Ubenimide competitively inhibits multiple aminopeptidases, including aminopeptidase B, aminopeptidase N, and leucine aminopeptidase. Aminopeptidases are involved in cell adhesion and tumor cell invasion. Therefore, inhibition of aminopeptidases may be part of the reason for urbenimide's antitumor effect. The drug can also activate T lymphocytes, macrophages, and bone marrow stem cells, and stimulate the release of interleukin-1 and interleukin-2, thereby further enhancing its antitumor activity.
Drug Indications Used as adjuvant therapy for acute and chronic myeloid leukemia, lung cancer, and nasopharyngeal carcinoma. It is also used to treat hypercholesterolemia. Ubenimex is a natural aminopeptidase inhibitor with a variety of biological activities, including immunomodulatory, antitumor and tissue protection [1][2][3][4] - Its mechanism of enhancing ATRA-induced NB4 cell differentiation involves inhibiting p38 MAPK phosphorylation, thereby blocking the negative regulatory effect of p38 on myeloid differentiation [2] - In diabetic retinopathy, Ubenimex exerts a protective effect by inhibiting retinal cell apoptosis, which may be related to the regulation of oxidative stress and inflammatory pathways [1] - The immunomodulatory activities of Ubenimex include enhancing humoral immunity in normal mice and reversing immunosuppression in cyclophosphamide (CY)-treated mice, making it a potential adjuvant for immunotherapy [3] - In Dictyostelium Bestatin (Ubenimex), produced by discoideum, inhibits cell growth and differentiation by targeting aminopeptidase, which is involved in cell cycle regulation and developmental signal transduction [4]. |
| Molecular Formula |
C16H24N2O4
|
|
|---|---|---|
| Molecular Weight |
308.37
|
|
| Exact Mass |
308.173
|
|
| CAS # |
58970-76-6
|
|
| Related CAS # |
Bestatin hydrochloride;65391-42-6;Bestatin trifluoroacetate;223763-80-2
|
|
| PubChem CID |
72172
|
|
| Appearance |
White to off-white solid powder
|
|
| Density |
1.2±0.1 g/cm3
|
|
| Boiling Point |
604.7±55.0 °C at 760 mmHg
|
|
| Melting Point |
245 °C (dec.)(lit.)
|
|
| Flash Point |
319.5±31.5 °C
|
|
| Vapour Pressure |
0.0±1.8 mmHg at 25°C
|
|
| Index of Refraction |
1.557
|
|
| LogP |
2.64
|
|
| Hydrogen Bond Donor Count |
4
|
|
| Hydrogen Bond Acceptor Count |
5
|
|
| Rotatable Bond Count |
8
|
|
| Heavy Atom Count |
22
|
|
| Complexity |
367
|
|
| Defined Atom Stereocenter Count |
3
|
|
| SMILES |
CC(C)C[C@@H](C(=O)O)NC(=O)[C@H]([C@@H](CC1=CC=CC=C1)N)O
|
|
| InChi Key |
VGGGPCQERPFHOB-RDBSUJKOSA-N
|
|
| InChi Code |
InChI=1S/C16H24N2O4/c1-10(2)8-13(16(21)22)18-15(20)14(19)12(17)9-11-6-4-3-5-7-11/h3-7,10,12-14,19H,8-9,17H2,1-2H3,(H,18,20)(H,21,22)/t12-,13+,14+/m1/s1
|
|
| Chemical Name |
(2S)-2-[[(2S,3R)-3-amino-2-hydroxy-4-phenylbutanoyl]amino]-4-methylpentanoic acid
|
|
| Synonyms |
|
|
| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
|
| 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) |
|
|||
|---|---|---|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 0.83 mg/mL (2.69 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 8.3 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 0.83 mg/mL (2.69 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 8.3 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: ≥ 0.83 mg/mL (2.69 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 | 3.2429 mL | 16.2143 mL | 32.4286 mL | |
| 5 mM | 0.6486 mL | 3.2429 mL | 6.4857 mL | |
| 10 mM | 0.3243 mL | 1.6214 mL | 3.2429 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
COA