| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
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| 10mg | |||
| Other Sizes |
| Targets |
c-di-AMP primarily targets the STING (STimulator of INterferon Genes) pathway in eukaryotic cells. It binds directly to the transmembrane protein STING with a Kd of ~4.59 µM, activating the TBK3-IRF3 signaling pathway and triggering the production of type I IFN and TNF. In bacteria, c-di-AMP regulates cell wall synthesis, potassium ion channels, DNA repair, and biofilm formation. It is also essential for cell growth, survival, and virulence of pathogenic bacteria including S. aureus, L. monocytogenes, S. pyogenes, and Mycobacterium spp.
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| ln Vitro |
For many Gram-positive bacteria, the manufacture of cell walls, potassium channels, DNA repair, and the formation of biofilms are all regulated by c-di-AMP (cyclic diadenylate-AMP) signaling. A number of well-known human pathogenic bacteria, such as Staphylococcus aureus, Listeria monocytogenes, Streptococcus pyogenes, and Mycobacterium spp., depend on c-di-AMP for cell growth, survival, and virulence. significant[1]. Cyclic diadenylate, or c-di-AMP, functions as a strong mucosal adjuvant by binding to model antigens like OVA or β-Gal and inducing humoral and cellular reactions [4].
c-di-AMP signaling is a central factor in many Gram-positive bacteria regulating cell wall synthesis, potassium ion channels, DNA repair, and biofilm formation. It is essential for cell growth, survival, and virulence of several human pathogenic bacteria including S. aureus, L. monocytogenes, S. pyogenes, and Mycobacterium spp. c-di-AMP combines with model antigens such as OVA or β-Gal and acts as a potent mucosal adjuvant stimulating both humoral and cellular responses. It enables dissection of the cGAS-independent STING axis in bacterial infection models. |
| ln Vivo |
c-di-AMP acts as a balanced Th1/Th2/Th17 mucosal adjuvant with superior dendritic cell activation compared to c-di-GMP for intranasal vaccine formulations. As a bacterial second messenger, it regulates cell growth, survival, and virulence primarily within Gram-positive bacteria and modulates host immune responses. Recombinant BCG with c-di-AMP as an endogenous adjuvant induces elevated immune responses after Mycobacterium tuberculosis infection. The combination vaccine adjuvant system Alum/c-di-AMP results in enhanced immune responses post immunization.
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| Enzyme Assay |
c-di-AMP binding assays involve measuring affinity for STING using surface plasmon resonance or isothermal titration calorimetry (Kd ~4.59 µM). STING activation is assessed by measuring TBK3-IRF3 signaling pathway activation and type I IFN production in cell-free systems. Radiolabeled c-di-AMP binding displacement studies may be performed to determine binding affinity and specificity. For bacterial studies, DisA diadenylate cyclase inhibitor screening assays are performed leveraging structural divergence from mammalian cGAS. Assays are performed in appropriate buffer systems with positive controls such as known STING agonists.
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| Cell Assay |
c-di-AMP cell-based assays are conducted in immune cells and cell lines expressing STING. Cells are cultured in appropriate media at 37°C with 5% CO2 and treated with c-di-AMP at varying concentrations. STING pathway activation is assessed by measuring type I IFN and TNF production by ELISA. IRF3 phosphorylation is analyzed by Western blot. For dendritic cell activation studies, cells are treated with c-di-AMP and activation markers are assessed by flow cytometry. For bacterial studies, cultures of Gram-positive bacteria are treated with c-di-AMP and cell growth, survival, and virulence are assessed. Experiments are performed in triplicate with appropriate controls.
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| Animal Protocol |
c-di-AMP in vivo studies are conducted in mouse models of infection and vaccination. For vaccine adjuvant studies, animals are immunized with antigens combined with c-di-AMP via intranasal or other routes. Immune responses are assessed by measuring antibody titers, cytokine production, and T-cell responses. For infection models, animals are challenged with Mycobacterium tuberculosis or other pathogens and bacterial burden in tissues is quantified. Survival rates and clinical signs are monitored. Studies are conducted in accordance with institutional animal care guidelines.
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| ADME/Pharmacokinetics |
c-di-AMP (MW 658.4 g/mol, C20H24N10O12P2) is a cyclic dinucleotide with moderate solubility in water (3.33 mg/mL, 5.06 mM, need ultrasonic). It is supplied as a lyophilized powder with ≥98% purity (HPLC). The compound is stable as a powder at -20°C for up to 3 years and in solvent at -80°C for 6 months. Pharmacokinetic properties such as half-life, bioavailability, and tissue distribution would be determined in species-specific studies. c-di-AMP serves as an analytical standard for DisA diadenylate cyclase inhibitor screening.
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| Toxicity/Toxicokinetics |
c-di-AMP is generally well-tolerated in preclinical studies at doses used for vaccine adjuvant and immunomodulation research. The compound is a naturally occurring bacterial second messenger and has been evaluated for safety in various animal models. As a STING agonist, it may elicit inflammatory responses at high doses. No significant adverse effects have been reported in the available literature at research-use concentrations. The compound is intended for research use only. Standard safety precautions should be followed when handling. Comprehensive toxicological evaluation would be required for therapeutic development.
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| References |
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| Additional Infomation |
Cyclic adenosine diphosphate (c-di-AMP) is a cyclic purine dinucleotide, a 3',5'-cyclic dimer of adenosine monophosphate (AMP). It is a metabolite of Mycoplasma genitalium. It is a cyclic purine dinucleotide and adenosine monophosphate ribonucleotide. It is the conjugate acid of cyclic adenosine diphosphate (c-di-AMP).
c-di-AMP is a naturally occurring bacterial cyclic dinucleotide second messenger and a potent STING agonist with a Kd of ~4.59 µM. It regulates bacterial cell wall synthesis, potassium channels, DNA repair, and biofilm formation. c-di-AMP acts as a potent mucosal adjuvant stimulating humoral and cellular responses and enables dissection of the cGAS-independent STING axis. It serves as an analytical standard for DisA diadenylate cyclase inhibitor screening. All applications are limited to non-human research use. |
| Molecular Formula |
C20H24N10O12P2
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|---|---|
| Molecular Weight |
658.411960000001
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| Exact Mass |
658.105
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| CAS # |
54447-84-6
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| Related CAS # |
c-di-AMP disodium;2734909-87-4;c-di-AMP diammonium
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| PubChem CID |
11158091
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| Appearance |
White to off-white solid powder
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| LogP |
-5.1
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| Hydrogen Bond Donor Count |
6
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| Hydrogen Bond Acceptor Count |
20
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
44
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| Complexity |
1080
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| Defined Atom Stereocenter Count |
8
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| SMILES |
O[C@H]([C@@](OP(O)(OC[C@](O[C@@H](N1C2=NC=NC(N)=C2N=C1)[C@@H]3O)([H])[C@@]3([H])O4)=O)([H])[C@](COP4(O)=O)([H])O5)[C@@H]5N6C7=NC=NC(N)=C7N=C6
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| InChi Key |
PDXMFTWFFKBFIN-XPWFQUROSA-N
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| InChi Code |
InChI=1S/C20H24N10O12P2/c21-15-9-17(25-3-23-15)29(5-27-9)19-11(31)13-7(39-19)1-37-43(33,34)42-14-8(2-38-44(35,36)41-13)40-20(12(14)32)30-6-28-10-16(22)24-4-26-18(10)30/h3-8,11-14,19-20,31-32H,1-2H2,(H,33,34)(H,35,36)(H2,21,23,25)(H2,22,24,26)/t7-,8-,11-,12-,13-,14-,19-,20-/m1/s1
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| Chemical Name |
(1S,6R,8R,9R,10S,15R,17R,18R)-8,17-bis(6-aminopurin-9-yl)-3,12-dihydroxy-3,12-dioxo-2,4,7,11,13,16-hexaoxa-3λ5,12λ5-diphosphatricyclo[13.3.0.06,10]octadecane-9,18-diol
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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) |
H2O : ~3.33 mg/mL (~5.06 mM)
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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 | 1.5188 mL | 7.5941 mL | 15.1881 mL | |
| 5 mM | 0.3038 mL | 1.5188 mL | 3.0376 mL | |
| 10 mM | 0.1519 mL | 0.7594 mL | 1.5188 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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