| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
IC50: ＞1.0 μM (A1AT)[1]
A1AT modulator 2 targets alpha-1 antitrypsin (A1AT), a serine protease inhibitor (serpin) that primarily inhibits neutrophil elastase (NE). A1AT deficiency is a genetic disorder that leads to chronic lung and liver disease. This modulator aims to enhance or correct the dysfunctional activity of A1AT, thereby restoring its protective function. By influencing the protein's function, this modulator could offer a targeted approach to managing A1AT deficiency and improving patient outcomes. |
|---|---|
| ln Vitro |
In vitro, A1AT modulator 2 modulates the activity of alpha-1 antitrypsin (A1AT). It has an IC50 value greater than 1.0 microM and an EC50 value less than 0.4 microM. These values indicate that the compound is an effective modulator, with high potency (low EC50) for its enhancing effect, and low potential for off-target inhibition (IC50 > 1.0 microM). The specific details of the assay used to determine these values are not provided, but it likely measures the ability of A1AT to inhibit neutrophil elastase (NE). The compound can be used for research in infection and inflammation.
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| ln Vivo |
A1AT modulator 2 is designed to be used in animal models of infection and inflammation to study the therapeutic potential of modulating A1AT function. By enhancing the anti-elastase activity of A1AT, the compound may reduce tissue damage caused by excessive neutrophil elastase activity during inflammatory states, such as chronic obstructive pulmonary disease (COPD), acute respiratory distress syndrome (ARDS), or other inflammatory conditions. It can be used for research in infection and inflammation, with potential therapeutic applications for A1AT deficiency.
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| Enzyme Assay |
A biochemical assay for A1AT modulators is a neutrophil elastase (NE) inhibition assay using purified human neutrophil elastase and a chromogenic or fluorogenic substrate (e.g., MeOSuc-Ala-Ala-Pro-Val-AMC). The assay is performed in a 96-well plate. A fixed concentration of A1AT (or A1AT treated with the modulator) is pre-incubated with varying concentrations of the test compound for 30 minutes at 37degC. Then, NE is added, followed by the substrate. The increase in fluorescence (excitation 360 nm, emission 460 nm) is measured over time. The half-maximal effective concentration (EC50) for enhancement of A1AT activity is calculated.
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| Cell Assay |
An in vitro cellular assay for A1AT modulation uses human lung epithelial cells (e.g., A549) or primary human neutrophils. Cells are treated with an inflammatory stimulus (e.g., lipopolysaccharide, LPS) to induce neutrophil elastase (NE) release or inflammation. The test compound (A1AT modulator 2) is added to the culture medium (0.1-10 microM). After 24-48 hours, cell damage is assessed by measuring lactate dehydrogenase (LDH) release. NE activity in the culture supernatant can be measured using a fluorescent substrate. The levels of pro-inflammatory cytokines (e.g., IL-6, IL-8, TNF-alpha) are measured by ELISA. The compound's ability to reduce NE activity and cell damage is assessed.
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| Animal Protocol |
In vivo studies for A1AT modulator 2 can be performed in a mouse model of acute lung inflammation (e.g., lipopolysaccharide (LPS)-induced lung injury). Mice are treated with A1AT modulator 2 at doses of 1-10 mg/kg via oral gavage or intraperitoneal (IP) injection, 1-2 hours before intratracheal administration of LPS (5 microg/mouse). After 6-24 hours, bronchoalveolar lavage fluid (BALF) is collected. Endpoints include neutrophil count in BALF, total protein (as a measure of vascular leak), and levels of pro-inflammatory cytokines (IL-6, TNF-alpha) and neutrophil elastase activity in BALF. Lung tissue is collected for histopathological analysis (H&E staining) and myeloperoxidase (MPO) activity.
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| ADME/Pharmacokinetics |
A1AT modulator 2 has a molecular weight of 455.48 and a molecular formula of C27H22FN3O3. It is a solid powder with a purity of ≥98%. It is soluble in DMSO (e.g., 10 mg/mL). For in vivo use, it can be formulated in vehicles such as 10% DMSO, 40% PEG300, 5% Tween-80, and 45% saline, or 10% DMSO, 90% corn oil, to achieve a clear solution at 2-5 mg/mL. The compound is stable as a powder at -20degC for up to 3 years, and in solution at -80degC for up to 1 year. Specific pharmacokinetic parameters (half-life, bioavailability) are not reported.
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| Toxicity/Toxicokinetics |
Specific toxicity data for A1AT modulator 2 is not available. As a modulator of a human protein, it is generally designed to have an acceptable safety profile. The IC50 value of greater than 1.0 microM indicates low off-target enzyme inhibition, suggesting that the compound may have a favorable safety margin. However, for research use, standard safety precautions should be followed, including the use of personal protective equipment (gloves, lab coat, goggles) and working in a well-ventilated area. The compound is not for human therapeutic use.
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| References |
[1]. Bandarage, Upul Keerthi, et al. Pyrrolo[2,3-f]indazole derivatives as modulators of α-1 antitrypsin and their preparation. US20200361939. 2020.
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| Additional Infomation |
A1AT modulator 2 (compound 33; CAS 2555004-05-0) is a small molecule modulator of alpha-1 antitrypsin (A1AT). Alpha-1 antitrypsin (A1AT) is a serine protease inhibitor (serpin) encoded by the SERPINA1 gene. A1AT deficiency (AATD) is an autosomal co-dominant disorder that predisposes patients to chronic obstructive pulmonary disease (COPD) and liver disease. Current treatments include augmentation therapy with purified human A1AT; however, this modulator aims to enhance the endogenous dysfunctional protein's activity. This compound is strictly for research use in infection and inflammation studies and is not approved for clinical use.
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| Molecular Formula |
C27H22FN3O3
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|---|---|
| Molecular Weight |
455.480289936066
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| Exact Mass |
455.164
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| CAS # |
2555004-05-0
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| PubChem CID |
155285167
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| Appearance |
Typically exists as solid at room temperature
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| LogP |
5
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
4
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| Heavy Atom Count |
34
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| Complexity |
718
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C1COCCC1C2=C(C3=C(N2C4=CC=C(C=C4)F)C=C5C=NNC5=C3)C6=CC=C(C=C6)C(=O)O
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| InChi Key |
BEMKXFFRSZUYSK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C27H22FN3O3/c28-20-5-7-21(8-6-20)31-24-13-19-15-29-30-23(19)14-22(24)25(26(31)17-9-11-34-12-10-17)16-1-3-18(4-2-16)27(32)33/h1-8,13-15,17H,9-12H2,(H,29,30)(H,32,33)
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| Chemical Name |
4-[5-(4-fluorophenyl)-6-(oxan-4-yl)-1H-pyrrolo[2,3-f]indazol-7-yl]benzoic 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 |
| 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 (109.77 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.49 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 (5.49 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. 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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.1955 mL | 10.9774 mL | 21.9549 mL | |
| 5 mM | 0.4391 mL | 2.1955 mL | 4.3910 mL | |
| 10 mM | 0.2195 mL | 1.0977 mL | 2.1955 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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