| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Protein Arginine Deiminase 2 (PAD2)[1]
PAD2 (Protein Arginine Deiminase 2). |
|---|---|
| ln Vitro |
Compound 32a, or AFM32a hydrochloride, has an EC 50 of 8.3 μM in HEK293T/PAD2 cells in the target engagement experiment. This indicates that AFM32a's improved potency outweighs its relatively weak cell entry ability[1]. With an EC 50 of 2.7 μM, AFM32a (compound 32a; 1-25 μM) hydrochloride treatment substantially suppresses histone H3 citrullination in HEK293T/PAD2 cells[1].
AFM32a hydrochloride exhibits an EC50 of 8.3 microM for PAD2 target engagement in HEK293T cells. It substantially suppresses histone H3 citrullination (EC50: 2.7 microM), demonstrating potent inhibition of PAD2 enzymatic activity in a cellular context. |
| ln Vivo |
Specific in vivo data for AFM32a is limited as it is a research tool. PAD2 inhibitors are generally studied in models of autoimmune diseases (e.g., rheumatoid arthritis) and cancers. Their in vivo efficacy is measured by reduction in citrullination levels in target tissues.
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| Enzyme Assay |
A non-cellular fluorometric assay is conducted. Recombinant human PAD2 is incubated with a substrate peptide (e.g., BAEE) and CaCl2 in reaction buffer. AFM32a is pre-incubated with the enzyme. The reaction is initiated by adding substrate. The production of citrulline is measured by a colorimetric method using diacetyl monoxime.
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| Cell Assay |
HEK293T cells overexpressing PAD2 are seeded in culture plates. Cells are treated with varying concentrations of AFM32a (1-25 microM) for 4-6 hours. Cells are harvested, lysed, and protein lysates are subjected to Western blotting using an anti-citrulline antibody (e.g., F95) to measure H3 citrullination levels.
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| Animal Protocol |
A mouse model of autoimmune arthritis is established (e.g., collagen-induced arthritis, CIA). AFM32a is administered via intraperitoneal (IP) injection daily. Clinical arthritis scores (joint swelling) are measured, and joint tissue is harvested for histopathological analysis and measurement of citrullination.
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| ADME/Pharmacokinetics |
The pharmacokinetic profile is not well-defined. As a small molecule inhibitor, it is likely suitable for oral or IP administration in animals. It is designed to be cell-permeable to reach its intracellular target, PAD2. Specific data on half-life, clearance, or bioavailability is not available.
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| Toxicity/Toxicokinetics |
No specific toxicology data is available for this research compound. PAD2 is primarily expressed in hematopoietic cells. Given its role in inflammatory gene regulation, chronic inhibition of PAD2 could potentially lead to immunosuppression or impact immune cell development. This is a research tool only.
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| References | |
| Additional Infomation |
PAD enzymes catalyze the conversion of arginine residues to citrulline (citrullination) in target proteins. PAD2 is implicated in the pathogenesis of rheumatoid arthritis (RA), multiple sclerosis (MS), and lupus as well as various cancers. AFM32a is a selective chemical probe for investigating PAD2 biology.
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| Molecular Formula |
C25H30CLFN6O3
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|---|---|
| Molecular Weight |
517
|
| CAS # |
2988594-85-8
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| Related CAS # |
AFM32a;2095109-82-1
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| Appearance |
White to off-white solid powder
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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 (e.g. under nitrogen), avoid exposure to moisture and light. |
| 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 (~193.42 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (4.84 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 (4.84 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 (4.84 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 | 1.9342 mL | 9.6712 mL | 19.3424 mL | |
| 5 mM | 0.3868 mL | 1.9342 mL | 3.8685 mL | |
| 10 mM | 0.1934 mL | 0.9671 mL | 1.9342 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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