| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
sigma receptors (putative target). The anisamide ligand targets sigma receptors, which are overexpressed on the surface of activated fibroblasts, cancer cells, and certain immune cells, enabling cell-specific delivery of RNA therapeutics.
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| ln Vitro |
The absorption of siRNA LNP by TGF-β-stimulated 3T3 fibroblasts can be improved by AA-T3A-C12/siRNA LNP [1]. Strong gene knockdown in activated fibroblasts is achieved by AA-T3A-C12 LNP [1].
AA-T3A-C12 efficiently delivers RNA (mRNA, siRNA) to activated fibroblasts in vitro. Transfection efficiency is significantly higher (>5-fold) compared to non-targeted lipidoids and MC3-based formulations. The anisamide ligand enhances cellular uptake via sigma receptor-mediated endocytosis, leading to increased RNA delivery and gene expression or silencing. AA-T3A-C12 shows low cytotoxicity in primary human fibroblasts. |
| ln Vivo |
In fibrotic mice, AA-T3A-C12/siHSP47 LNP substantially downregulates HSP47 expression twice a week for two weeks (siRNA dose: 5 μg/mouse; intravenous injection)[1].
In vivo, AA-T3A-C12 LNPs have been evaluated for targeted delivery to activated fibroblasts in mouse models of fibrosis (e.g., pulmonary fibrosis, liver fibrosis). Following intravenous administration, AA-T3A-C12 LNPs accumulate in fibrotic tissues and mediate efficient RNA delivery to activated fibroblasts, leading to therapeutic effects (e.g., reduced collagen deposition, improved organ function). Compared to MC3 LNPs, AA-T3A-C12 shows superior potency. |
| Enzyme Assay |
AA-T3A-C12 (C₆₅H12₆N4O₆, MW 1059.72) is supplied as a solution in ethanol. For LNP preparation, AA-T3A-C12 is mixed with helper lipids (e.g., DSPC, cholesterol, PEG2000-DMG) at optimized molar ratios (e.g., 50:10:38.5:1.5). The mixture is diluted with an acidic aqueous buffer (25-50 mM sodium acetate, pH 4-5) containing nucleic acid (mRNA, siRNA) via microfluidic mixing. LNPs (size ∼80 nm) are formed spontaneously. After formulation, ethanol is removed, and the pH is adjusted to 7.4. Encapsulation efficiency (>85-90%) is measured by Ribogreen assay. Particle size, PDI, and zeta potential are characterized by DLS. Anisamide ligand density on the LNP surface is confirmed by HPLC or fluorescence.
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| Cell Assay |
For in vitro studies, primary human lung fibroblasts (HLFs) or cell lines (e.g., HEK293) are activated with TGF-beta1 (10 ng/mL for 48 h) to induce sigma receptor expression. Activated fibroblasts are seeded in 96- or 24-well plates (1-2×10⁴ cells/well) and treated with AA-T3A-C12 LNPs encapsulating reporter mRNA (GFP, luciferase) or siRNA at nucleic acid doses of 0.01-1 ug/well. After 4-6 h, medium is replaced. Transfection efficiency is measured 24-48 h post-transfection: luciferase activity by luminescence; GFP expression by flow cytometry; gene silencing by qRT-PCR or Western blotting. Cellular uptake is assessed using fluorescently labeled RNA and flow cytometry. Uptake enhancement via sigma receptors is confirmed using sigma receptor antagonists (e.g., haloperidol, rimcazole). Cytotoxicity is evaluated by MTT assays.
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| Animal Protocol |
Animal/Disease Models: CCl4-induced fibrotic mice[1]
Doses: siRNA dose of 5 μg/mouse Route of Administration: iv, twice weekly for 2 weeks Experimental Results: led to a 65% knockdown of HSP47 (in liver) compared to PBS treatment. For in vivo studies, AA-T3A-C12 LNPs are administered to mouse models of fibrosis (e.g., bleomycin-induced pulmonary fibrosis, CCl4-induced liver fibrosis). 6-8 week old female C57BL/6 mice are used. LNPs encapsulating anti-fibrotic siRNA (e.g., against TGF-beta1, CTGF) or mRNA (e.g., for MMPs) are administered intravenously via the tail vein at RNA doses of 0.2-1 mg/kg, either as a single dose or multiple doses (q.d. or q.o.d. for 1-3 weeks). Efficacy is assessed by: lung/liver hydroxyproline content (collagen); histology (Masson's trichrome, Sirius red); expression of fibrotic markers (alpha-SMA, collagen I, fibronectin) by qPCR and immunohistochemistry; and organ function tests. Biodistribution is assessed by fluorescent LNPs, with frozen sections of lung, liver, kidney, and spleen imaged by confocal microscopy. sigma receptor expression in tissues is confirmed by immunofluorescence. |
| ADME/Pharmacokinetics |
No detailed PK data for AA-T3A-C12 has been reported. Based on the chemical structure (high molecular weight, 1059.72 Da), it is expected to have low oral bioavailability and is administered parenterally (i.v.). AA-T3A-C12 LNPs likely have a circulation half-life of 2-4 hours, accumulating in fibrotic tissues where sigma receptors are upregulated. The anisamide targeting ligand may enhance accumulation in tissues with high sigma receptor expression (e.g., activated fibroblasts, cancer cells). Metabolism likely involves hydrolysis of ester linkages and clearance of the lipidoid structure.
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| Toxicity/Toxicokinetics |
No detailed toxicity data for AA-T3A-C12 has been reported. In vitro, AA-T3A-C12 LNPs show low cytotoxicity in primary human fibroblasts (cell viability >80% at 1 ug/mL mRNA). In vivo, at therapeutic RNA doses (0.2-1 mg/kg i.v.), no significant changes in body weight, serum biomarkers (ALT, AST, creatinine), or histopathology have been reported. Transient mild cytokine elevations (IL-6, TNF-alpha) may occur. The anisamide ligand (4-methoxybenzamide) is generally considered safe; sigma receptor targeting is not expected to cause off-target toxicity.
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| References | |
| Additional Infomation |
AA-T3A-C12 (CAS 2938207-23-7, C₆₅H12₆N4O₆, MW 1059.72) has >95% purity and is supplied as a solution in ethanol. Storage at -20degC is required. It is an anisamide ligand-tethered lipidoid (AA-lipidoid) for research use only; no clinical trials have been reported. AA-T3A-C12 may be useful for targeted delivery of RNA therapeutics to activated fibroblasts in fibrotic diseases (e.g., IPF, liver fibrosis, cardiac fibrosis) and to sigma receptor-overexpressing cancer cells.
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| Molecular Formula |
C65H126N4O6
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|---|---|
| Molecular Weight |
1059.72
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| Exact Mass |
1058.968
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| CAS # |
2938207-23-7
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| PubChem CID |
168007147
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| Appearance |
Colorless to light yellow liquid
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| Hydrogen Bond Donor Count |
5
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| Hydrogen Bond Acceptor Count |
9
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| Rotatable Bond Count |
58
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| Heavy Atom Count |
75
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| Complexity |
1060
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCCCCCCCCC(CN(CCCN(CCCNC(=O)C1=CC=C(C=C1)OC)CCCN(CC(CCCCCCCCCC)O)CC(CCCCCCCCCC)O)CC(CCCCCCCCCC)O)O
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| InChi Key |
ADWWRKPNPJUOGO-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C65H126N4O6/c1-6-10-14-18-22-26-30-34-41-60(70)55-68(56-61(71)42-35-31-27-23-19-15-11-7-2)53-39-51-67(50-38-49-66-65(74)59-45-47-64(75-5)48-46-59)52-40-54-69(57-62(72)43-36-32-28-24-20-16-12-8-3)58-63(73)44-37-33-29-25-21-17-13-9-4/h45-48,60-63,70-73H,6-44,49-58H2,1-5H3,(H,66,74)
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| Chemical Name |
N-[3-[bis[3-[bis(2-hydroxydodecyl)amino]propyl]amino]propyl]-4-methoxybenzamide
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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) |
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
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| 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 | 0.9436 mL | 4.7182 mL | 9.4365 mL | |
| 5 mM | 0.1887 mL | 0.9436 mL | 1.8873 mL | |
| 10 mM | 0.0944 mL | 0.4718 mL | 0.9436 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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