Sodium c-di-AMP (cyclic diadenylate) is a STING agonist that binds to STING, thereby activating the TBK3-IRF3 signaling pathway and triggering the production of type I IFN and TNF. c-di-AMP sodium is a bacterial second-generation messenger primarily in Gram-positive (Gram+) cells that regulates cell growth and ultimately virulence. c-di-AMP (cyclic diadenylate) also regulates the next step in the immune response. c-di-AMP sodium works as a potent mucosal adjuvant, stimulating humoral and cellular responses

Physicochemical Properties

Molecular Formula	C20H25N10NAO12P2
Molecular Weight	682.409595251083
Exact Mass	702.068
CAS #	2734909-87-4
Related CAS #	c-di-AMP diammonium;c-di-AMP;54447-84-6
PubChem CID	122714672
Appearance	White to off-white solid powder
Hydrogen Bond Donor Count	4
Hydrogen Bond Acceptor Count	20
Rotatable Bond Count	2
Heavy Atom Count	46
Complexity	1070
Defined Atom Stereocenter Count	8
SMILES	O[C@@H]1[C@]2([H])OP(OC[C@@]3([H])O[C@@H](N4C=NC5=C(N=CN=C45)N)[C@H](O)[C@]3([H])OP(O)(=O)OC[C@@]2([H])O[C@H]1N1C=NC2=C(N=CN=C12)N)(O)=O.[NaH]
InChi Key	IXWPEIRAKLMJQW-VEQUCWRQSA-L
InChi Code	InChI=1S/C20H24N10O12P2.2Na/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);;/q;2*+1/p-2/t7-,8-,11-,12-,13-,14-,19-,20-;;/m1../s1
Chemical Name	disodium;(1S,6R,8R,9R,10S,15R,17R,18R)-8,17-bis(6-aminopurin-9-yl)-3,12-dioxido-3,12-dioxo-2,4,7,11,13,16-hexaoxa-3λ5,12λ5-diphosphatricyclo[13.3.0.06,10]octadecane-9,18-diol
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 Note: (1). This product requires protection from light (avoid light exposure) during transportation and storage.(2). Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture.
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

In 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) sodium signaling. A number of well-known human pathogenic bacteria, such as Staphylococcus aureus, Listeria monocytogenes, Streptococcus pyogenes, and Mycobacterium spp., depend on sodium c-di-AMP for cell growth, survival, and virulence. It's quite significant [1]. When combined with model antigens like OVA or β-Gal, c-di-AMP sodium functions as a strong mucosal adjuvant that stimulates cellular and humoral responses [4].

References

[1]. c-di-AMP: An Essential Molecule in the Signaling Pathways that Regulate the Viability and Virulenceof Gram-Positive Bacteria. Genes (Basel). 2017 Aug 7;8(8). [2]. Recombinant BCG With Bacterial Signaling Molecule Cyclic di-AMP as Endogenous AdjuvantInduces Elevated Immune Responses After Mycobacterium tuberculosis Infection. Front Immunol. 2019 Jul 3;10:1519. [3]. The Combination Vaccine Adjuvant System Alum/c-di-AMP Results in Quantitative and QualitativeEnhanced Immune Responses Post Immunization. Front Cell Infect Microbiol. 2019 Feb 19;9:31. [4]. Intranasal delivery of influenza rNP adjuvanted with c-di-AMP induces strong humoral and cellularimmune responses and provides protection against virus challenge. PLoS One. 2014 Aug 20;9(8):e104824.

Solubility Data

Solubility (In Vitro)

DMSO : ~270 mg/mL (~384.41 mM) H2O : ≥ 50 mg/mL (~71.19 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 6.75 mg/mL (9.61 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 67.5 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: ≥ 6.75 mg/mL (9.61 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 67.5 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. Solubility in Formulation 3: ≥ 6.75 mg/mL (9.61 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 67.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.4654 mL	7.3270 mL	14.6539 mL
	5 mM	0.2931 mL	1.4654 mL	2.9308 mL
	10 mM	0.1465 mL	0.7327 mL	1.4654 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.