| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| Other Sizes |
| Targets |
IC50: 28 nM (PARL)[1]
PARL (presenilin‑associated rhomboid‑like protein), a mitochondrial inner membrane rhomboid protease. PARL is involved in the processing of PINK1, a key regulator of mitophagy. PARL-IN-1 inhibits PARL, leading to the stabilization of PINK1 on the outer mitochondrial membrane and activation of the PINK1/Parkin pathway. |
|---|---|
| ln Vitro |
In cell‑free enzymatic assays, PARL-IN-1 TFA inhibits PARL activity with an IC50 of 28 nM. The inhibition leads to the accumulation of PINK1 on the outer mitochondrial membrane and the subsequent recruitment of Parkin, which ubiquitinates mitochondrial proteins and tags damaged mitochondria for autophagic removal (mitophagy).
|
| ln Vivo |
In vivo, PARL-IN-1 TFA can be used to study the role of mitophagy in cellular and animal models of Parkinson‘s disease. By activating the PINK1/Parkin pathway, it promotes the clearance of damaged mitochondria, which is a protective mechanism against neurodegeneration. The in vivo efficacy of PARL-IN-1 TFA in disease models is an active area of research.
|
| Enzyme Assay |
A general cell‑free protocol for assessing PARL inhibition: A fluorometric protease activity assay is used. Recombinant PARL (or a PARL‑containing mitochondrial membrane fraction) is incubated with a fluorogenic peptide substrate (e.g., a peptide corresponding to the PINK1 cleavage site) in a buffer containing varying concentrations of PARL-IN-1 TFA (0.1‑1000 nM) at 37degC for 30‑60 minutes. The reaction is stopped, and the fluorescence (due to cleavage of the peptide) is measured. The IC50 is calculated from the inhibition curve.
|
| Cell Assay |
A general cellular protocol for PARL-IN-1 TFA: HEK293T or SH‑SY5Y neuroblastoma cells are seeded in 6‑well plates. Cells are treated with PARL-IN-1 TFA at concentrations of 0.1, 1, 5, 10, 25, 50 uM for 24 hours. Mitophagy is assessed by measuring the co‑localization of LC3B (autophagosome marker) with mitochondrial markers (e.g., Tom20) using immunofluorescence microscopy. The levels of PINK1 and Parkin recruitment to mitochondria are assessed by Western blot after mitochondrial fractionation. The degradation of mitochondrial proteins (e.g., Tom20, COX1) is measured by Western blot in the presence or absence of lysosomal inhibitors (e.g., chloroquine).
|
| Animal Protocol |
A general animal protocol for PARL-IN-1 TFA: A mouse model of Parkinson‘s disease (e.g., MPTP‑treated mice or PINK1/Parkin knockout mice) is used. Male C57BL/6 mice are treated with MPTP (20 mg/kg, 4 times at 2‑hour intervals) to induce dopaminergic neuron degeneration. PARL-IN-1 TFA is formulated in a suitable vehicle and administered via intraperitoneal (IP) injection at doses of 5, 15, and 30 mg/kg daily for 7 days prior to and following MPTP administration. The mice are euthanized, and the striatum is dissected. Tyrosine hydroxylase (TH) levels are measured by Western blot and immunohistochemistry to assess dopaminergic neuron survival. Mitophagy markers (PINK1, Parkin, LC3B) are assessed by Western blot. Behavioral tests (pole test, rotarod) are performed to assess motor function.
|
| ADME/Pharmacokinetics |
General PK protocol for PARL-IN-1 TFA: Male Sprague‑Dawley rats are administered PARL-IN-1 TFA via IP (10 mg/kg) and IV (2 mg/kg). Blood samples are collected at 0.083, 0.25, 0.5, 1, 2, 4, 8, 12, and 24 hours. Plasma concentrations are quantified by LC‑MS/MS. PK parameters (Cmax, Tmax, AUC, t1/2, clearance, Vd, and IP bioavailability) are calculated.
|
| Toxicity/Toxicokinetics |
General toxicity protocol for PARL-IN-1 TFA: A 14‑day repeated‑dose IP toxicity study is performed in ICR mice. PARL-IN-1 TFA is administered at doses of 5, 15, and 40 mg/kg/day. Parameters include clinical signs, body weight, food consumption, hematology (CBC), serum chemistry (ALT, AST, BUN, creatinine), and histopathology of major organs (liver, kidney, spleen, heart, lung, brain). Special attention is paid to the brain and liver due to the role of mitophagy in these organs.
|
| References | |
| Additional Infomation |
PARL-IN-1 TFA is the TFA salt form of PARL-IN-1. The molecular weight and formula of the free base are not provided. The TFA salt is used to improve solubility. PARL is a rhomboid serine protease located in the inner mitochondrial membrane. Its natural substrate is PINK1. PARL cleaves PINK1, leading to its proteasomal degradation and preventing the accumulation of PINK1 on damaged mitochondria. Inhibition of PARL stabilizes PINK1, promoting mitophagy. PARL-IN-1 TFA is a valuable research tool for studying mitophagy and its role in Parkinson‘s disease.
|
| Molecular Formula |
C41H57F3N6O9
|
|---|---|
| Molecular Weight |
834.92
|
| Appearance |
Solid powder
|
| 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: (1). This product requires protection from light (avoid light exposure) during transportation and storage. (2). Please store this product in a sealed and protected environment (e.g. under nitrogen), 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) |
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 | 1.1977 mL | 5.9886 mL | 11.9772 mL | |
| 5 mM | 0.2395 mL | 1.1977 mL | 2.3954 mL | |
| 10 mM | 0.1198 mL | 0.5989 mL | 1.1977 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
