| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| 10mg |
|
||
| 25mg |
|
||
| 50mg |
|
||
| 100mg |
|
||
| 250mg | |||
| 500mg | |||
| Other Sizes |
Purity: ≥98%
AA26-9 is a potent and broad-spectrum serine hydrolase inhibitor. AA26-9-inhibited enzymes originated from diverse functional subclasses of serine hydrolases, including lipases/phospholipases, thioesterases, and uncharacterized enzymes.
| Targets |
Serine hydrolases (SHs) including AADACL1, ABHD6, ABHD11, ABHD13, APEH, BAT5, CTSA, ESD, FAAH, LYPLA1, LYPLA2, LYPLA3, PAFAH2, PRCP [1]
|
|---|---|
| ln Vitro |
Based on a piperazine scaffold that was previously demonstrated to inhibit serine hydrolase in the presence of p-nitrophenoxycarbamate, AA26-9 was synthesized. Enzymes deriving from various functional subclasses of serine hydrolases are inhibited by AA26-9: these include peptidases (APEH, PRCP, CTSA), lypases/phospholipases (AADACL1, ABHD6, ESD, FAAH, PAFAH2, LYPLA3), thioesterases (LYPLA1, LYPLA2), and unidentified enzymes (ABHD11, ABHD13, BAT5). LYPLA1, one of AA26-9's enzymatic targets, is inhibited by the enzyme through covalent carbamylation of its serine nucleophile (S114). Thirty percent of the over forty serine hydrolases present in T cells are inhibited by AA26-9 [1].
In living mouse BW5147 T‑cell hybridoma cells, treatment with AA26-9 at 20 μM for 4 h resulted in inhibition of 15 serine hydrolases (greater than 75% inhibition as determined by ABPP‑SILAC). These included AADACL1, ABHD6, ABHD11, ABHD13, APEH, BAT5, CTSA, ESD, FAAH, LYPLA1, LYPLA2, LYPLA3, PAFAH2, and PRCP. [1] In vitro treatment of mouse T‑cell proteomes (soluble and membrane fractions) with AA26-9 at 1 μM for 30 min inhibited several SHs, including ABHD11, APEH, FAAH, PAFAH2, and LYPLA1, as visualized by competitive gel‑based ABPP using the FP‑Rh probe. [1] Mass spectrometry analysis confirmed that AA26-9 inhibits LYPLA1 by covalent carbamoylation of the enzyme’s serine nucleophile (S114). [1] AA26-9 showed inhibitory activity against approximately one‑third of the more than 40 serine hydrolases detected in immortalized T‑cell lines. [1] Compared to the structurally related carbamate AA38‑3, AA26-9 inhibited a substantially larger number of SHs (15 vs. 3) in mouse T‑cells. [1] |
| Enzyme Assay |
Competitive activity‑based protein profiling (ABPP) using the serine hydrolase‑directed probe FP‑rhodamine (FP‑Rh) was performed. Mouse brain membrane proteome or mouse T‑cell lysates were incubated with AA26-9 (20 μM for cell lysates or 1 μM for in vitro proteome) for 30 min at 37 °C, followed by labeling with FP‑Rh (2 μM, 30 min, 25 °C). Samples were separated by SDS‑PAGE, and FP‑Rh‑labeled proteins were detected by in‑gel fluorescence scanning. [1]
For quantitative analysis, competitive ABPP‑SILAC (stable isotope labeling of amino acids in culture) was used. Isotopically “light” and “heavy” mouse T‑cells were treated with AA26-9 (20 μM) or DMSO, respectively, for 4 h. Cells were lysed, proteomes were labeled with FP‑biotin (5 μM, 90 min), mixed at 1:1 ratio, enriched with avidin, digested on‑bead with trypsin, and analyzed by LC‑MS/MS on an LTQ‑Orbitrap instrument. Light/heavy ratios of tryptic peptides were quantified to determine enzyme inhibition. [1] To confirm the covalent modification mechanism, recombinant LYPLA1 was analyzed by MS after treatment with AA26-9, revealing carbamoylation of the catalytic serine (S114). [1] |
| Cell Assay |
Mouse BW5147 T‑cell hybridoma cells were cultured in medium containing AA26-9 at 20 μM or DMSO (control) for 4 h. After treatment, cells were harvested, lysed, separated into soluble and membrane fractions, and analyzed by competitive gel‑based ABPP using FP‑Rh probe (2 μM, 30 min). Inhibition of serine hydrolases was detected by in‑gel fluorescence scanning. [1]
For ABPP‑SILAC, mouse T‑cells were cultured under “light” (12C6-14N2-lysine and 12C6-14N4-arginine) or “heavy” (13C6-15N2-lysine and 13C6-15N4-arginine) conditions. Heavy cells were treated with DMSO, light cells with AA26-9 (20 μM, 4 h). Cells were then lysed, labeled with FP‑biotin, combined, and processed for LC‑MS/MS as described. [1] |
| References | |
| Additional Infomation |
AA26-9 is a broad‑spectrum 1,2,3‑triazole urea inhibitor that irreversibly inactivates serine hydrolases via carbamoylation of the catalytic serine nucleophile. It serves as a lead scaffold for the development of selective inhibitors for individual SHs, as demonstrated by the optimization into potent and selective probes such as AA74‑1 (APEH inhibitor), AA39‑2 (PAFAH2 inhibitor), and AA44‑2 (ABHD11 inhibitor). [1]
AA26-9 inhibits serine peptidases, lipases, amidases, and esterases/thioesterases, indicating broad reactivity across major subgroups of the serine hydrolase superfamily. [1] The compound was synthesized via a two‑step click chemistry procedure: substituted alkynes were reacted with in situ‑formed azidomethanol to yield 4‑substituted triazoles, which were then carbamoylated to give triazole urea products. AA26-9 contains a piperidine carbamoyl group and an unsubstituted 1,2,3‑triazole leaving group. [1] |
| Molecular Formula |
C7H10N4O
|
|---|---|
| Molecular Weight |
166.184
|
| Exact Mass |
166.085
|
| Elemental Analysis |
C, 50.59; H, 6.07; N, 33.71; O, 9.63
|
| CAS # |
1312782-34-5
|
| Related CAS # |
1312782-34-5;
|
| PubChem CID |
46829239
|
| Appearance |
White to off-white solid powder
|
| Density |
1.4±0.1 g/cm3
|
| Boiling Point |
306.7±25.0 °C at 760 mmHg
|
| Flash Point |
139.3±23.2 °C
|
| Vapour Pressure |
0.0±0.7 mmHg at 25°C
|
| Index of Refraction |
1.691
|
| LogP |
-0.78
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
3
|
| Rotatable Bond Count |
0
|
| Heavy Atom Count |
12
|
| Complexity |
178
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
O=C(N1N=NC=C1)N2CCCC2
|
| InChi Key |
IFXGAMVQLDJRBQ-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C7H10N4O/c12-7(10-4-1-2-5-10)11-6-3-8-9-11/h3,6H,1-2,4-5H2
|
| Chemical Name |
Pyrrolidin-1-yl(triazol-1-yl)methanone
|
| Synonyms |
AA 26-9 AA-26-9 AA26-9 AA 269 AA-269 AA269
|
| 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) |
DMSO : ~100 mg/mL (~601.76 mM)
H2O : ≥ 100 mg/mL (~601.76 mM) |
|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (15.04 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 (15.04 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 (15.04 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 | 6.0176 mL | 30.0879 mL | 60.1757 mL | |
| 5 mM | 1.2035 mL | 6.0176 mL | 12.0351 mL | |
| 10 mM | 0.6018 mL | 3.0088 mL | 6.0176 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
COA
