| Size | Price | Stock | Qty |
|---|---|---|---|
| 100mg |
|
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| 500mg | |||
| 1g | |||
| Other Sizes |
| Targets |
- Angiotensin I-converting enzyme (ACE) (target for antihypertensive-related activity): Ki = 5.2 μM [1]
- Insulin signaling pathway proteins (target for insulin sensitivity regulation): Including insulin receptor substrate 1 (IRS-1) and glucose transporter 4 (GLUT4) [2] |
|---|---|
| ln Vitro |
H-Val-Pro-Pro-OH, a derivative of proline peptide, has an IC50 of 9 μM and can inhibit the angiotensin I-converting enzyme (ACE) [1]. H-Val-Pro-Pro-OH can prevent insulin resistance and increase 3T3-F442A preadipocytes' sensitivity to insulin. H-Val-Pro-Pro-OH also possesses anti-inflammatory and anti-hypertensive qualities. H-Val-Pro-Pro-OH helps adipocytes express the glucose transporter 4 (GLUT4) and improves glucose uptake in adipocytes treated with TNF [2].
- ACE inhibitory activity: H-Val-Pro-Pro-OH (VPP) exhibited competitive inhibitory activity against angiotensin I-converting enzyme (ACE) in vitro. Its inhibition constant (Ki) was 5.2 μM. When the concentration of H-Val-Pro-Pro-OH increased from 1 μM to 10 μM, the inhibitory rate of ACE activity increased from 28% to 72%, showing a concentration-dependent inhibitory effect [1] - Insulin sensitivity enhancement: In 3T3-F442A preadipocytes induced to insulin resistance (by high glucose and high insulin treatment), H-Val-Pro-Pro-OH (10–100 μM) significantly enhanced insulin sensitivity. At a concentration of 50 μM, it increased insulin-stimulated glucose uptake by 32% compared with the insulin-resistant control group. It also upregulated the protein expression of GLUT4 (by 45%) and the phosphorylation level of IRS-1 (by 38%) in the cells, while reducing the expression of pro-inflammatory cytokine TNF-α (by 29%) [2] - Adipocyte differentiation regulation: H-Val-Pro-Pro-OH (50 μM) promoted the differentiation of 3T3-F442A preadipocytes into mature adipocytes, as evidenced by a 35% increase in the number of lipid droplets (stained by Oil Red O) and a 28% increase in the expression of adipocyte differentiation marker protein PPAR-γ [2] |
| Enzyme Assay |
- ACE activity inhibition assay: The reaction system (total volume 1 mL) contained 50 mM borate buffer (pH 8.3), 300 mM NaCl, 5 mM hippuryl-His-Leu (substrate for ACE), and different concentrations of H-Val-Pro-Pro-OH. The reaction was initiated by adding ACE enzyme solution, incubated at 37°C for 30 min, and terminated by adding 1 M HCl. The amount of hippuric acid produced (a product of substrate hydrolysis) was measured spectrophotometrically at 228 nm. The inhibitory rate of ACE activity was calculated using the formula: Inhibitory rate (%) = [1 - (OD sample - OD blank)/(OD control - OD blank)] × 100. The Ki value was determined by Lineweaver-Burk double reciprocal plotting [1]
|
| Cell Assay |
- Insulin resistance model establishment and glucose uptake assay: 3T3-F442A preadipocytes were cultured in DMEM medium containing 10% fetal bovine serum. To induce insulin resistance, cells were treated with 25 mM glucose and 100 nM insulin for 48 h. Then, the cells were divided into groups: control group (normal medium), insulin-resistant group (high glucose + high insulin), and H-Val-Pro-Pro-OH treatment groups (10, 50, 100 μM). After 24 h of treatment, cells were incubated with 2-deoxy-[³H]-D-glucose (radioactive glucose analog) for 30 min. The radioactivity in the cells was measured using a liquid scintillation counter to calculate glucose uptake capacity [2]
- Western blot assay for protein expression: After treatment with H-Val-Pro-Pro-OH, 3T3-F442A cells were lysed with RIPA buffer containing protease and phosphatase inhibitors. The protein concentration was determined by BCA method. Equal amounts of protein were separated by SDS-PAGE and transferred to PVDF membranes. The membranes were blocked with 5% skim milk, incubated with primary antibodies against GLUT4, IRS-1, p-IRS-1 (Tyr632), and β-actin (internal reference) overnight at 4°C, then incubated with secondary antibody for 1 h at room temperature. The protein bands were visualized using an enhanced chemiluminescence (ECL) system, and the band intensity was quantified using ImageJ software [2] - qPCR assay for gene expression: Total RNA was extracted from H-Val-Pro-Pro-OH-treated 3T3-F442A cells using TRIzol reagent. RNA was reverse-transcribed into cDNA using a reverse transcription kit. qPCR was performed using SYBR Green PCR Master Mix, with primers specific for TNF-α, PPAR-γ, and GAPDH (internal reference). The relative gene expression levels were calculated using the 2⁻ΔΔCt method [2] |
| References |
|
| Additional Infomation |
Val-Pro-Pro is a tripeptide composed of L-valine and two L-proline units linked by peptide bonds. It is a metabolite that is functionally related to L-valine and L-proline. Val-Pro-Pro has been reported to exist in Trypanosoma brucei, and there is relevant data. - Source of isolation: H-Val-Pro-Pro-OH (VPP) is a milk-derived tripeptide. It was originally isolated from yogurt fermented by lactic acid bacteria. In reference [1], it was purified from the peptides of yogurt by ultrafiltration (molecular weight cutoff of 3 kDa), ion exchange chromatography and reversed-phase high performance liquid chromatography (RP-HPLC) [1]. - ACE inhibition mechanism: H-Val-Pro-Pro-OH acts as a competitive inhibitor of ACE. It binds to the active site of ACE and competes with the natural substrate (angiotensin I) for binding sites, thereby inhibiting the ACE-mediated conversion of angiotensin I to angiotensin II (a potent vasoconstrictor) and the degradation of bradykinin (a vasodilator), which may help it exert its antihypertensive effect [1].
- Insulin sensitivity regulation mechanism: H-Val-Pro-Pro-OH enhances the insulin sensitivity of 3T3-F442A preadipocytes through multiple pathways: 1) upregulating GLUT4 expression to promote glucose transport into cells; 2) increasing IRS-1 phosphorylation to activate downstream insulin signaling pathways; 3) reducing TNF-α expression to alleviate insulin resistance caused by inflammation; 4) promoting adipocyte differentiation to improve lipid metabolism [2]. |
| Molecular Formula |
C15H25N3O4
|
|---|---|
| Molecular Weight |
311.3767
|
| Exact Mass |
311.185
|
| CAS # |
58872-39-2
|
| Related CAS # |
H-Val-Pro-Pro-OH TFA;2828433-08-3
|
| PubChem CID |
9818200
|
| Appearance |
White to off-white solid powder
|
| Density |
1.275 g/cm3
|
| Boiling Point |
566.446ºC at 760 mmHg
|
| Flash Point |
296.376ºC
|
| LogP |
0.612
|
| Hydrogen Bond Donor Count |
2
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
4
|
| Heavy Atom Count |
22
|
| Complexity |
466
|
| Defined Atom Stereocenter Count |
3
|
| SMILES |
CC(C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N2CCC[C@H]2C(=O)O)N
|
| InChi Key |
DOFAQXCYFQKSHT-SRVKXCTJSA-N
|
| InChi Code |
InChI=1S/C15H25N3O4/c1-9(2)12(16)14(20)17-7-3-5-10(17)13(19)18-8-4-6-11(18)15(21)22/h9-12H,3-8,16H2,1-2H3,(H,21,22)/t10-,11-,12-/m0/s1
|
| Chemical Name |
(2S)-1-[(2S)-1-[(2S)-2-amino-3-methylbutanoyl]pyrrolidine-2-carbonyl]pyrrolidine-2-carboxylic acid
|
| 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) |
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
|
|---|---|
| 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 | 3.2115 mL | 16.0576 mL | 32.1151 mL | |
| 5 mM | 0.6423 mL | 3.2115 mL | 6.4230 mL | |
| 10 mM | 0.3212 mL | 1.6058 mL | 3.2115 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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