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| 10mg |
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| Other Sizes |
| Targets |
Fis1 (mitochondrial fission protein 1). P110 is a peptide inhibitor that disrupts the interaction between Drp1 and Fis1, thereby inhibiting excessive mitochondrial fragmentation.
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| ln Vitro |
In cell-free binding assays, P110 specifically binds to Fis1, blocking the Fis1-Drp1 interaction. This reduces pathological mitochondrial fragmentation without interfering with the physiological function of Drp1 in mitochondrial division. P110 reduces mitochondrial fragmentation and mitochondrial ROS production in a mouse model of Parkinson‘s disease. The peptide inhibits the Fis1-Drp1 interaction with an IC₅0 value in the low micromolar range. It reduces programmed cell death and improves cell viability by protecting mitochondrial integrity.
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| ln Vivo |
In vivo, P110 reduces pathological functions in numerous models of neurodegeneration (such as Parkinson's and Alzheimer‘s disease), ischemia (cerebral and myocardial), and sepsis without blocking the physiological functions of Drp1. It reduces mitochondrial fragmentation and mitochondrial ROS production in a mouse model of Parkinson's disease. P110 has anti-inflammatory, immunomodulatory, mitochondrial protective, and neuroprotective activities.
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| Enzyme Assay |
A general cell-free protocol for assessing the Drp1-Fis1 interaction and its inhibition by P110: A fluorescence polarization (FP) competitive binding assay is used. Recombinant GST-tagged Fis1 protein (50 nM) is incubated with a fluorescently labeled Drp1 peptide (corresponding to the Drp1 binding site for Fis1, labeled with 5-FAM) in an assay buffer (50 mM Tris-HCl, pH 7.4, 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 0.01% Tween-20) at room temperature for 30 minutes. Varying concentrations of P110 (0.1 nM to 100 uM) are added. The fluorescence polarization signal is measured using a microplate reader. The IC₅0 is calculated from the competition curve. Alternatively, a surface plasmon resonance (SPR) assay can be used. Fis1 protein is immobilized on a sensor chip, and a Drp1 peptide is flowed over the chip in the presence or absence of P110. The association and dissociation rates are measured to calculate the Kd for the interaction and the inhibitory constant Ki for P110.
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| Cell Assay |
A general cellular protocol for assessing P110-mediated mitochondrial protection: Primary neurons or immortalized cell lines (e.g., SH-SY5Y, HeLa, or H9c2 cardiomyocytes) are seeded in 12-well plates at 2×10⁵ cells/well. The cells are treated with various concentrations of P110 (0.1, 1, 5, 10, 25 uM) for 4 hours prior to exposure to a cellular stressor that induces mitochondrial fragmentation. The stressor can be rotenone (1 uM, an inhibitor of complex I, for 24 hours) to model Parkinson‘s disease, oxygen-glucose deprivation (OGD) to model ischemia, or LPS (1 ug/mL) to model sepsis. After the stressor exposure, the cells are incubated with the mitochondrial dye MitoTracker Red CMXRos (100 nM) for 30 minutes at 37degC. Cells are fixed with 4% paraformaldehyde and stained with DAPI. Images are captured using a confocal or fluorescence microscope. Mitochondrial morphology is assessed by classifying mitochondria as tubular, intermediate, or fragmented. For quantitative analysis, the mitochondrial fragmentation index or the aspect ratio (major axis/minor axis) is calculated using ImageJ software. For apoptosis analysis, cells are stained with Annexin V-FITC and PI and analyzed by flow cytometry. Mitochondrial ROS production is measured using MitoSOX Red. ATP levels are measured using a luciferase-based ATP assay kit.
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| Animal Protocol |
General animal protocol for evaluating neuroprotective effects in a Parkinson's disease model: Male C57BL/6 mice (8-10 weeks old, 20-25 g) are used. The mice are randomly divided into treatment groups (n=10 per group). P110 is formulated in sterile PBS or saline and administered via intraperitoneal (IP) injection at doses of 1, 3, and 10 mg/kg once daily for 7-14 days. A vehicle control group (PBS) and a positive control group (e.g., rasagiline or selegiline) are included. Parkinson‘s disease is induced by intraperitoneal injection of MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine, 20 mg/kg) four times at 2-hour intervals on day 1. P110 is administered 1 hour before the first MPTP injection and then daily thereafter. Behavioral tests are performed 7 days after MPTP administration. The pole test measures the time it takes for a mouse to turn and descend a vertical pole. The rotarod test measures the latency to fall from a rotating rod (accelerating from 4 to 40 rpm over 5 minutes). At the end of the study, the mice are euthanized, and the brains are harvested. The striatum is dissected, and tissue homogenates are prepared. Tyrosine hydroxylase (TH) levels are measured by Western blot and immunohistochemistry as a marker of dopaminergic neuron integrity. Mitochondrial morphology in the substantia nigra is examined by transmission electron microscopy (TEM).
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| ADME/Pharmacokinetics |
General pharmacokinetic protocol for P110: P110 is a heptapeptide with a molecular weight of 756.85 Da. Peptides typically have poor oral bioavailability and a short half-life due to rapid proteolysis. A general pharmacokinetic protocol would involve administering P110 to mice via intravenous (IV, 1 mg/kg) and intraperitoneal (IP, 10 mg/kg) injection. Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-dose. Plasma concentrations are quantified by LC-MS/MS. PK parameters (Cmax, Tmax, AUC, t½, clearance, Vd, and IP bioavailability) are calculated.
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| Toxicity/Toxicokinetics |
General toxicity protocol for P110: A 14-day repeated-dose toxicity study is performed in ICR mice. P110 is administered via intraperitoneal (IP) injection at doses of 5, 15, and 50 mg/kg/day for 14 consecutive days. Clinical signs, body weight, and food consumption are monitored daily. At the end of the study, blood samples are collected for hematology (complete blood count with differential) and serum chemistry (ALT, AST, ALP, BUN, creatinine, total protein, albumin, glucose, CPK). Gross necropsy is performed, and the weights of major organs (liver, kidney, spleen, heart, lung, brain, testes) are recorded. Histopathological examination of these organs is conducted.
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| References | |
| Additional Infomation |
P110 is a cell-permeable heptapeptide that specifically inhibits the Drp1-Fis1 interaction. It binds to Fis1, preventing Drp1 from being recruited to the mitochondrial outer membrane. P110 has a sequence that is not provided in the available data. The peptide has anti-inflammatory, immunomodulatory, mitochondrial protective, and neuroprotective activities. P110 reduces mitochondrial fragmentation and ROS production, decreases programmed cell death, and improves cell viability. The peptide has been studied in models of neurodegeneration (Parkinson‘s disease, Alzheimer's disease), ischemia (stroke, myocardial infarction), and sepsis. P110 is a valuable research tool for studying the role of Drp1-Fis1-mediated mitochondrial fission in various disease conditions.
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| Molecular Formula |
C32H56N10O11
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|---|---|
| Molecular Weight |
756.85
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| Exact Mass |
756.413
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| CAS # |
1411976-18-5
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| PubChem CID |
171902142
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| Appearance |
Solid powder
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| Density |
1.47±0.1 g/cm3(Temp: 20 °C; Press: 760 Torr)(predicted)
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| LogP |
0
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| Hydrogen Bond Donor Count |
11
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| Rotatable Bond Count |
23
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| Heavy Atom Count |
53
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| Complexity |
1340
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| Defined Atom Stereocenter Count |
6
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| SMILES |
CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC(=O)O)N
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| InChi Key |
VUAKYKPDARUALO-LLINQDLYSA-N
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| InChi Code |
InChI=1S/C32H56N10O11/c1-16(2)11-20(40-26(47)18(33)13-25(45)46)28(49)41-21(12-17(3)4)30(51)42-10-6-8-23(42)29(50)39-19(7-5-9-36-32(34)35)27(48)37-14-24(44)38-22(15-43)31(52)53/h16-23,43H,5-15,33H2,1-4H3,(H,37,48)(H,38,44)(H,39,50)(H,40,47)(H,41,49)(H,45,46)(H,52,53)(H4,34,35,36)/t18-,19-,20-,21-,22-,23-/m0/s1
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| Chemical Name |
(3S)-3-amino-4-[[(2S)-1-[[(2S)-1-[(2S)-2-[[(2S)-1-[[2-[[(1S)-1-carboxy-2-hydroxyethyl]amino]-2-oxoethyl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]carbamoyl]pyrrolidin-1-yl]-4-methyl-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-4-oxobutanoic acid
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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 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)
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| Solubility (In Vitro) |
DMSO : 50 mg/mL (66.06 mM; with sonication)
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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 | 1.3213 mL | 6.6063 mL | 13.2127 mL | |
| 5 mM | 0.2643 mL | 1.3213 mL | 2.6425 mL | |
| 10 mM | 0.1321 mL | 0.6606 mL | 1.3213 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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