Cyclic-di-GMP disodium is a STING agonist and bacterial second messenger that coordinates different aspects of bacterial growth and behavior such as motility, virulence, biofilm formation, and cell cycle progression. Cyclic-di-GMP disodium has anti-cancer cell proliferation/growth activity and can also induce increased CD4 receptor expression and cell cycle arrest. Cyclic-di-GMP disodium may be utilized in cancer research.

Physicochemical Properties

Molecular Formula	C20H25N10NAO14P2
Molecular Weight	714.408395528793
Exact Mass	734.058
CAS #	2222132-40-1
Related CAS #	Cyclic-di-GMP diammonium;609343-82-0;Cyclic-di-GMP;61093-23-0;Cyclic-di-GMP sodium
PubChem CID	137120247
Appearance	Off-white to light yellow solid powder
Hydrogen Bond Donor Count	6
Hydrogen Bond Acceptor Count	18
Rotatable Bond Count	2
Heavy Atom Count	48
Complexity	1320
Defined Atom Stereocenter Count	8
SMILES	O=C1N=C(N)NC2N([C@@H]3O[C@]4([H])COP(O[C@@]5([H])[C@@H](O)[C@H](N6C=NC7C(N=C(N)NC6=7)=O)O[C@]5([H])COP(O)(=O)O[C@@]4([H])[C@H]3O)(O)=O)C=NC1=2.[NaH]
InChi Key	SVIXOIGPVFFABR-XDODWTFASA-L
InChi Code	InChI=1S/C20H24N10O14P2.2Na/c21-19-25-13-7(15(33)27-19)23-3-29(13)17-9(31)11-5(41-17)1-39-45(35,36)44-12-6(2-40-46(37,38)43-11)42-18(10(12)32)30-4-24-8-14(30)26-20(22)28-16(8)34;;/h3-6,9-12,17-18,31-32H,1-2H2,(H,35,36)(H,37,38)(H3,21,25,27,33)(H3,22,26,28,34);;/q;2*+1/p-2/t5-,6-,9-,10-,11-,12-,17-,18-;;/m1../s1
Chemical Name	disodium;2-amino-9-[(1S,6R,8R,9R,10S,15R,17R,18R)-17-(2-amino-6-oxo-1H-purin-9-yl)-9,18-dihydroxy-3,12-dioxido-3,12-dioxo-2,4,7,11,13,16-hexaoxa-3λ5,12λ5-diphosphatricyclo[13.3.0.06,10]octadecan-8-yl]-1H-purin-6-one
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

Targets	STING[1][2][3][4].
ln Vitro	Human colon cancer cells cannot proliferate when exposed to cyclic-di-GMP disodium (0.5-50 μM; 5 days)[1]. ?In Jurkat cells, cyclic-di-GMP disodium (0.5-50 μM; 5 days) selectively increases CD4 expression[2]. ?In Jurkat cells, cyclic-di-GMP disodium (0.5-50 μM; 5 days) causes cell cycle arrest at the S-phase[2].
ln Vivo	The administration of two consecutive vaccinations, separated by nine days, with 100 μg/per of cyclic-di-GMP disodium (iv) improves the immune responses TriVax elicits in mice against melanoma and amplifies its anti-tumor effects[3].
Cell Assay	Cell Proliferation Assay[1] Cell Types: H508 cells Tested Concentrations: 0.5- 50 µM Incubation Duration: 5 days Experimental Results: diminished basal H508 cell proliferation by approx 15%, even inhibited acetylcholine- and EGF-induced cell proliferation. Cell Viability Assay[2] Cell Types: Jurkat cells Tested Concentrations: 50 µM Incubation Duration: 24 h Experimental Results: Specifically induced of CD4 (no effect on the expression of CD8), with a 6.3-fold upregulation over control and in a dose-dependent manner. Cell Cycle Analysis[2] Cell Types: Jurkat cells Tested Concentrations: 50 µM Incubation Duration: 24 h Experimental Results: Increased the percentage of cells in S-phase by 79%, with almost complete disappearance of G2/M-phase cells which diminished by 93%.
Animal Protocol	Animal/Disease Models: C57BL/ 6 (B6) mice (8- to 10weeks old)[3]. Doses: 100 µg/per Route of Administration: intravenous (iv) injection; two sequential vaccinations 9 days apart; combine with TriVax. Experimental Results: Dramatically higher numbers of antigen-specific CD8 T cells when combined with TriVax. (TriVax consisted of a mixture of 120 μg Pam-hgp100, 100 μg hgp100 or 100 μg Ova, 50 or 25 μg anti-CD40 antibody, and 25 μg Poly-IC). Enhanced the anti- tumor activity of TriVax.
References	[1]. 3',5'-Cyclic diguanylic acid (c-di-GMP) inhibits basal and growth factor-stimulated human colon cancer cell proliferation. Biochem Biophys Res Commun. 2005 Apr 1;329(1):40-5. [2]. Elevated expression of the CD4 receptor and cell cycle arrest are induced in Jurkat cells by treatment with the novel cyclic dinucleotide 3',5'-cyclic diguanylic acid. FEBS Lett. 1999 Feb 5;444(1):125-9. [3]. STING activator c-di-GMP enhances the anti-tumor effects of peptide vaccines in melanoma-bearing mice. Cancer Immunol Immunother. 2015 Aug;64(8):1057-66. [4]. Cyclic di-GMP: second messenger extraordinaire. Nat Rev Microbiol. 2017 May;15(5):271-284.

Solubility Data

Solubility (In Vitro)	H2O: 160 mg/mL (217.87 mM)
Solubility (In Vivo)	Solubility in Formulation 1: 50 mg/mL (68.09 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with sonication.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.3998 mL	6.9988 mL	13.9976 mL
	5 mM	0.2800 mL	1.3998 mL	2.7995 mL
	10 mM	0.1400 mL	0.6999 mL	1.3998 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.