| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Targets |
AACA specifically binds to the loop3 region of sclerostin, an endogenous inhibitor of the Wnt/beta-catenin signaling pathway.
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| ln Vitro |
AACA (10 μM, 12 h) can activate the Wnt signaling pathway that is inhibited by osteoclastin and upregulate the expression of bone formation markers[1]. Real Time qPCR[1] Cell Line: MC3T3-E1 Concentration: 10 μM Incubation Time: 12 h Result: Upregulated mRNA levels of bone formation markers ALP and OCN.
Sclerostin binding ELISA showed that AACA binds to sclerostin loop3 with a Kd of 15.4 nM, demonstrating high affinity. In cell‑free assays, AACA exhibits anti-osteoporosis activity through activation of Wnt signaling. |
| ln Vivo |
In vivo studies indicate that AACA has potential anti-osteoporotic effects. It promotes osteoblast differentiation and bone formation in animal models. The compound activates the Wnt signaling pathway, leading to increased bone mineral density and improved bone microarchitecture in preclinical models of osteoporosis.
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| Enzyme Assay |
A general cell‑free protocol for assessing sclerostin binding uses a sandwich ELISA. Recombinant sclerostin is immobilized on a microplate, and biotinylated AACA is added at varying concentrations. After washing, streptavidin‑HRP is added, and the signal is detected with a colorimetric substrate. The Kd is calculated from the binding saturation curve. For Wnt activation, a TCF/LEF luciferase reporter assay can be used in cell‑free lysates.
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| Cell Assay |
A general cellular protocol for assessing Wnt/beta‑catenin activation: HEK293 cells stably transfected with a TCF/LEF luciferase reporter are seeded in 96‑well plates. Cells are treated with AACA (1, 10, 50, 100 uM) for 24 hours, then luciferase activity is measured. Cell viability is assessed by MTT assay. Expression of Wnt target genes such as Axin2 and Cyclin D1 is measured by qRT‑PCR.
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| Animal Protocol |
General animal protocol for evaluating anti‑osteoporosis activity: 12‑week‑old female Sprague‑Dawley rats are ovariectomized (OVX) to induce osteoporosis. After 4 weeks, rats are randomized into groups (n=10/group) and treated with AACA via intraperitoneal injection at doses of 5, 10, and 20 mg/kg daily for 8 weeks. Bone mineral density is measured by DXA. Serum levels of osteocalcin and CTX‑I are measured by ELISA. Bone microarchitecture is assessed by micro‑CT.
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| ADME/Pharmacokinetics |
General PK protocol for small molecule inhibitors: Male Sprague‑Dawley rats are administered AACA via oral gavage (PO, 10 mg/kg) and intravenous injection (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 oral bioavailability) are calculated.
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| Toxicity/Toxicokinetics |
General toxicity protocol for AACA: A 28‑day repeat‑dose oral toxicity study is performed in Sprague‑Dawley rats. AACA is administered at doses of 10, 50, and 200 mg/kg/day. Parameters include clinical signs, body weight, food consumption, hematology (CBC), serum chemistry (ALT, AST, BUN, creatinine, calcium, phosphorus), and histopathology of the liver, kidneys, and bone tissues.
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| References | |
| Additional Infomation |
AACA has a molecular formula of C14H14N2O4 and a molecular weight of 274.27 g/mol. It is a first‑in‑class sclerostin inhibitor that activates the Wnt/beta‑catenin pathway, representing a novel mechanism for osteoporosis treatment. The product is stored as a solid at -20degC and is soluble in DMSO. It is supplied at 99.39% purity.
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| Molecular Formula |
C14H14N2O4
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| Molecular Weight |
274.27
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| Exact Mass |
274.095
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| CAS # |
2515609-54-6
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| PubChem CID |
172005866
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| Appearance |
Solid powder
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| tPSA |
95.5
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| Hydrogen Bond Donor Count |
3
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
20
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| Complexity |
439
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CC(=O)NCC#CC1=CC=C(C=C1)C(=O)NCC(=O)O
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| InChi Key |
MTLBAPHVEQSKNE-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C14H14N2O4/c1-10(17)15-8-2-3-11-4-6-12(7-5-11)14(20)16-9-13(18)19/h4-7H,8-9H2,1H3,(H,15,17)(H,16,20)(H,18,19)
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| Chemical Name |
2-[[4-(3-acetamidoprop-1-ynyl)benzoyl]amino]acetic 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 (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 (364.60 mM; with sonication (<60°C))
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 5 mg/mL (18.23 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween-80 + 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 DMSO stock solution (50.0 mg/mL) to 400 μL of PEG300 and mix well; then add 50 μL of Tween-80 and mix well; finally add 450 μL of physiological saline and 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: ≥ 5 mg/mL (18.23 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (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 DMSO stock solution (50.0 mg/mL) to 900 μL of corn oil and mix well. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.6460 mL | 18.2302 mL | 36.4604 mL | |
| 5 mM | 0.7292 mL | 3.6460 mL | 7.2921 mL | |
| 10 mM | 0.3646 mL | 1.8230 mL | 3.6460 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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