| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| 50mg |
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| Other Sizes |
| Targets |
IC50: 4.5 nM (Hemoglobin-pepsin), 6.2 nM (Hemoglobin-proctase), 150 nM (Casein-pepsin), 260 nM (Hemoglobin-acid protease), 290 nM (Casein-proctase), 520 nM (Casein-acid protease)[1]
Pepstatin targets aspartic proteases by binding to their active sites. It exhibits high affinity for pepsin (Ki ~10-¹⁰ M), cathepsin D, and renin, as well as other acid proteases. The compound also shows affinity for microbial aspartic proteases and has been shown to compete with f-Met-Leu-Phe for binding to human neutrophils, suggesting an interaction with formyl peptide receptors. |
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| ln Vitro |
Pepstatin (Pepstatin A) (7 μM; 48 h) influences how HIV-specific gag protein is processed inside cells[2].
In vitro, Pepstatin A inhibits cathepsin D with sub-nanomolar potency but lacks selectivity. Pepstatin Trifluoroacetate analogues with trifluoromethylated modifications can achieve increased selectivity for CD over pepsin, with some (R)-TFM based analogues showing sub-nanomolar IC50 against CD. The compound also exhibits chemotactic activity and competes with f-Met-Leu-[3H]Phe for binding to human neutrophils. |
| ln Vivo |
Pepstatin, also known as Pepstatin A, is extremely nontoxic; its LD50s for mice, rats, rabbits, and dogs via the ip route are 1090 mg/kg, 875 mg/kg, 820 mg/kg, and 450 mg/kg, while for all species by the oral route they are > 2000 mg/kg1. The pylorus causes stomach ulcers in ligated Shay rats, although pepstatin (0.5–50 mg/kg, po) prevents this.
In vivo, Pepstatin derivatives have shown efficacy in animal models related to renin inhibition, effectively reducing blood pressure in dogs when administered at appropriate doses. The parent compound is poorly bioavailable, limiting its use in systemic applications. However, structural modifications continue to be evaluated for improved pharmacokinetic properties. |
| Enzyme Assay |
A typical enzyme inhibition assay for cathepsin D involves using a fluorescence resonance energy transfer (FRET) peptide substrate containing a fluorophore and quencher pair. The enzyme (e.g., 2.5 nM CD) is incubated with various concentrations of Pepstatin Trifluoroacetate (0.1-100 nM) in sodium acetate buffer (pH 4.5) at 37degC for 15 minutes. The substrate is then added, and fluorescence increase (excitation/emission = 340/490 nm) is monitored for 30 minutes. IC50 values are calculated by fitting data to a dose-response curve using nonlinear regression.
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| Cell Assay |
Cell viability assays are performed using cancer cell lines such as SKOV3 ovarian cancer cells. Cells are seeded in 96-well plates and treated with Pepstatin Trifluoroacetate (concentrations ranging from 1 nM to 100 uM) for 24-72 hours. Cytotoxicity is assessed using the MTT or CellTiter-Glo luminescent cell viability assay. Morphological changes and alterations in F-actin organization and fibronectin are evaluated by immunofluorescence staining.
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| Animal Protocol |
Animal/Disease Models: Pylorus ligated male Wistar rats[1]
Doses: 0.5, 1, 10 and 50 mg/kg Route of Administration: Oral administration, 15 minutes after pyloric ligation Experimental Results: Effectively prevent stomach ulceration. In rat or mouse models, Pepstatin is administered intraperitoneally at doses of 1-10 mg/kg to evaluate its effects on gastric acid secretion or kidney function (renin inhibition). Plasma renin activity (PRA) is measured by radioimmunoassay of generated angiotensin I. For tumor studies, mice bearing xenografts are treated with Pepstatin analogues to assess tumor growth inhibition. Detailed standard protocols for nonselective aspartic protease inhibitors apply. |
| ADME/Pharmacokinetics |
Pepstatin A suffers from poor oral bioavailability and rapid clearance from circulation. The compound is highly lipophilic and exhibits low aqueous solubility, which limits its systemic exposure when administered via oral or intravenous routes. TFA salt formulation and PEGylation strategies are sometimes employed to improve solubility and prolong circulation time, but significant pharmacokinetic optimization is generally required for in vivo applications.
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| Toxicity/Toxicokinetics |
Pepstatin has demonstrated low cytotoxicity at IC50 concentrations in cancer cell lines. However, some Pepstatin analogues have been observed to induce morphological changes in cells, including altered organization of F-actin and extracellular fibronectin. No significant acute toxicity has been reported in animal studies at moderate doses, though long-term toxicity data are limited for most analogues.
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| References |
[1]. Umezawa H, et al. Pepstatin, a new pepsin inhibitor produced by Actinomycetes. J Antibiot (Tokyo). 1970 May;23(5):259-62.
[2]. Seelmeier S, et al. Human immunodeficiency virus has an aspartic-type protease that can be inhibited by pepstatin A. Proc Natl Acad Sci U S A. 1988 Sep;85(18):6612-6. |
| Additional Infomation |
Pepstatin is strictly a biochemical research tool and has never received regulatory approval as a human therapeutic. It is widely used as a positive control in aspartic protease inhibition studies and as a matrix component in protease inhibitor cocktails for protein purification. The trifluoroacetate salt is the standard commercial form, providing superior handling and solubility compared to the free base. Pepstatin remains a critically important tool in enzymology to decipher the biological functions of cathepsins and renin.
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| Molecular Formula |
C36H64F3N5O11
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| Related CAS # |
Pepstatin;26305-03-3;Pepstatin Ammonium;Pepstatin acetate;28575-34-0
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| Appearance |
Typically exists as solid at room temperature
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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) |
DMSO :~100 mg/mL (~125.01 mM)
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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.) |
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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