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| Other Sizes |
| Targets |
Prolidase (also known as peptidase D), the enzyme responsible for cleaving iminodipeptides containing C-terminal proline or hydroxyproline. By inhibiting this enzyme, Carbobenzoxyproline mimics the enzymatic deficiency seen in PD patients, allowing researchers to study the downstream metabolic and cellular consequences.
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| ln Vitro |
Carphenoxyproline (6 mM; 0–10 days; pH=6.0) has been shown in long-term culturing of fibroblasts from patients with prolinase deficiency (PD) 33% to promote mitochondrial depolarization and accelerate cell death[1]. Fibroblast prolinase (FBP) is hydrolyzed by carphenoxyproline (0, 1, 3, 6 mM; 1 min; pH=6.0), exhibiting linear competitive inhibition [1].
In vitro, Carbobenzoxyproline (6 mM; 0-10 days; pH 6.0) causes mitochondrial depolarization and increases cellular death by 33% in long-term fibroblast cultures from PD patients. It also hydrolyzes fibroblast prolidase (FBP) in a linear competitive manner at concentrations of 0, 1, 3, and 6 mM. |
| ln Vivo |
In a mouse model, carphenoxyproline (60 mg/kg; injection; once daily; 3 weeks) inhibits erythrocyte prolinase in vivo [1].
In a mouse model, Carbobenzoxyproline (60 mg/kg; injection; once daily; 3 weeks) serves as an in vivo inhibitor of erythrocyte prolidase. This treatment results in a significant decrease in erythrocyte prolidase activity. |
| Enzyme Assay |
The in vitro enzyme inhibition assay typically involves incubating fibroblast prolidase (FBP) with varying concentrations of Carbobenzoxyproline (e.g., 0, 1, 3, 6 mM) at pH 6.0. The reaction is carried out for a short duration (e.g., 1 minute) to measure the initial rate of hydrolysis, demonstrating linear competitive inhibition kinetics.
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| Cell Assay |
The in vitro cell-based assay involves long-term culturing of fibroblasts derived from prolidase deficiency (PD) patients. Cells are treated with Carbobenzoxyproline (e.g., 6 mM) for up to 10 days at pH 6.0. The readout includes assessment of mitochondrial depolarization and quantification of cellular death.
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| Animal Protocol |
Animal/Disease Models: C57Bl/6J mice (4 weeks old) [1]
Doses: 60 mg/kg Route of Administration: injection; one time/day for 3 weeks. Experimental Results: Result in a significant decrease in erythrocyte prolinase activity. The in vivo study utilizes a C57Bl/6J mouse model (4 weeks old). Carbobenzoxyproline is administered via injection at a dose of 60 mg/kg once daily for a period of 3 weeks. The primary endpoint is the measurement of erythrocyte prolidase activity to confirm in vivo target inhibition. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties for Carbobenzoxyproline are not extensively detailed in the literature. As a small molecule (MW 249.26) with a logP of 1.17, it is expected to have moderate lipophilicity. The compound is typically administered via injection in animal models, suggesting potential issues with oral bioavailability.
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| Toxicity/Toxicokinetics |
Detailed toxicological data for Carbobenzoxyproline is not available in the standard research literature. As a research-grade compound, it is not intended for human use. In vitro studies at high concentrations (6 mM) have demonstrated cellular toxicity, including mitochondrial depolarization and increased cell death in fibroblast cultures.
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| References |
[1]. Lupi A, et al. N-benzyloxycarbonyl-L-proline: an in vitro and in vivo inhibitor of prolidase. Biochim Biophys Acta. 2005 Jun 30;1744(2):157-63.
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| Additional Infomation |
Carbobenzoxyproline is primarily a research tool for studying prolidase deficiency. It has not been developed as a therapeutic drug and is not approved for clinical use. The compound's mechanism of action involves competitive inhibition of prolidase, which helps researchers understand the metabolic basis of PD and test potential treatments in preclinical models.
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| Molecular Formula |
C13H15NO4
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|---|---|
| Molecular Weight |
249.26
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| Exact Mass |
249.1
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| CAS # |
1148-11-4
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| PubChem CID |
101987
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| Appearance |
White to off-white solid powder
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| Density |
1.3±0.1 g/cm3
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| Boiling Point |
432.3±45.0 °C at 760 mmHg
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| Melting Point |
76-78ºC
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| Flash Point |
215.3±28.7 °C
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| Vapour Pressure |
0.0±1.1 mmHg at 25°C
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| Index of Refraction |
1.582
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| LogP |
1.17
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
18
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| Complexity |
312
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| Defined Atom Stereocenter Count |
1
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| SMILES |
O=C(O)[C@H]1N(C(OCC2=CC=CC=C2)=O)CCC1
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| InChi Key |
JXGVXCZADZNAMJ-NSHDSACASA-N
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| InChi Code |
InChI=1S/C13H15NO4/c15-12(16)11-7-4-8-14(11)13(17)18-9-10-5-2-1-3-6-10/h1-3,5-6,11H,4,7-9H2,(H,15,16)/t11-/m0/s1
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| Chemical Name |
(2S)-1-phenylmethoxycarbonylpyrrolidine-2-carboxylic acid
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| Synonyms |
EINECS 214-557-4; Benzyloxycarbonylproline; Carbobenzoxyproline
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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 (~401.19 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (10.03 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 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL 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: ≥ 2.5 mg/mL (10.03 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 25.0 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: ≥ 2.5 mg/mL (10.03 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 | 4.0119 mL | 20.0594 mL | 40.1188 mL | |
| 5 mM | 0.8024 mL | 4.0119 mL | 8.0238 mL | |
| 10 mM | 0.4012 mL | 2.0059 mL | 4.0119 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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