| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
STING[1][2][3][4].
Cyclic-di-GMP diammonium directly binds to and activates the Stimulator of Interferon Genes (STING) protein, an adaptor protein in the cytosol that senses cyclic dinucleotides. Upon binding, STING dimerizes and translocates to the Golgi apparatus, where it activates TBK1 and IRF3 to induce type I interferons. This makes it a specific STING agonist for research. |
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| ln Vitro |
Human colon cancer cells cannot proliferate when exposed to cyclic-di-GMP diammonium (0.5-50 μM; 5 days) [1]. In Jurkat cells, cyclic-di-GMP diammonium (0.5-50 μM; 5 days) selectively upregulates CD4 expression [2]. In S phase cells, cyclic-di-GMP diammonium (0.5-50 μM; 5 days) causes Jurkat cell cycle arrest [2].
In cell-free assays, Cyclic-di-GMP diammonium binds directly to the STING protein with a Kd value of approximately 2-20 uM depending on the STING variant. It competes with natural ligand 2‘3'-cGAMP for the same binding pocket. Activation of STING by this compound leads to phosphorylation of TBK1 and IRF3 in a concentration-dependent manner. |
| ln Vivo |
TriVax stimulates an immunological response against melanoma in mice when administered intravenously with cyclic-di-GMP diammonium (100 μg/time); this further amplifies TriVax's anti-tumor impact[3].
In vivo, Cyclic-di-GMP diammonium is used as a vaccine adjuvant to enhance antigen-specific immune responses. It has been reported to enhance the anti-tumor effects of peptide vaccines in melanoma-bearing mice. Intratumoral injection of c-di-GMP can remodel the tumor microenvironment by promoting T-cell infiltration and reducing immunosuppressive cells. |
| Enzyme Assay |
The in vitro STING-binding protocol typically uses an isothermal titration calorimetry (ITC) or surface plasmon resonance (SPR) assay. Purified recombinant human STING protein (e.g., wild-type or common variants) is immobilized on a sensor chip. Cyclic-di-GMP diammonium is serially diluted (0.1-100 uM) and injected over the chip. Binding affinity (Kd) is calculated from the steady-state response or kinetic fitting.
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| 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%. For in vitro cellular assays, THP-1 human monocytic cells or primary human PBMCs are seeded in 96-well plates. Cells are treated with various concentrations of Cyclic-di-GMP diammonium (0.01-50 ug/mL) for 6-24 hours. Cell supernatants are harvested, and IFN-beta and CXCL10 levels are measured by ELISA. To confirm STING dependency, STING knockout cells or specific STING inhibitors are used as controls. |
| 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. In an in vivo animal model, C57BL/6 mice are implanted subcutaneously with B16-F10 melanoma cells. Once tumors reach 50-100 mm3, mice are intratumorally injected with Cyclic-di-GMP diammonium (25-100 ug in 50 uL PBS) on days 7, 10, and 13 post-implantation. Tumor volumes are measured every 2-3 days with calipers. On day 21, tumors are excised and digested for flow cytometry analysis of CD8+ T-cell infiltration. |
| ADME/Pharmacokinetics |
A standard in vivo animal protocol involves administration of Cyclic-di-GMP (5 mg/kg) into tumor-bearing mice at established intervals. This compound can be administered via various routes: intravenous injection (in PBS, pH 7.4, at 1-5 mg/kg), intratumoral injection (25-100 ug per tumor), or intramuscular administration (50-200 ug) for vaccine studies. Specific pharmacokinetic parameters for this compound have not been fully characterized.
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| Toxicity/Toxicokinetics |
The toxicological profile of Cyclic-di-GMP diammonium has not been extensively documented; however, as an endogenous bacterial signaling molecule and immune agonist, therapeutic indices are generally favorable at research doses. Systemic administration has been associated with transient cytokine release and flu-like symptoms. No significant organ toxicity or genotoxicity has been observed in animal models. Preclinical studies in mice using intratumoral injection up to 100 ug/dose showed no signs of acute toxicity or weight loss.
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| References | |
| Additional Infomation |
Cyclic-di-GMP diammonium is a research-grade chemical and is not approved for human therapeutic use. Its molecular formula is C20H30N12O14P2, with a molecular weight of 724.46 (free acid) and higher as diammonium salt. It is also a global bacterial second messenger regulating biofilm formation, motility, and virulence in diverse bacterial species. This compound is soluble in water and PBS, making it suitable for both in vitro and in vivo applications.
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| Molecular Formula |
C20H27N11O14P2
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|---|---|
| Molecular Weight |
707.44
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| Exact Mass |
724.147
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| CAS # |
609343-82-0
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| Related CAS # |
Cyclic-di-GMP disodium;2222132-40-1;Cyclic-di-GMP;61093-23-0;Cyclic-di-GMP sodium
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| PubChem CID |
162640464
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
8
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| Hydrogen Bond Acceptor Count |
18
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| Rotatable Bond Count |
2
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| Heavy Atom Count |
48
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| Complexity |
1320
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| Defined Atom Stereocenter Count |
8
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| 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.N
|
| InChi Key |
ORSUSHQNAQSNEL-XDODWTFASA-N
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| InChi Code |
InChI=1S/C20H24N10O14P2.2H3N/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);2*1H3/t5-,6-,9-,10-,11-,12-,17-,18-;;/m1../s1
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| Chemical Name |
diazanium;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
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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, 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)
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| Solubility (In Vitro) |
H2O: 50 mg/mL (69.02 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: 100 mg/mL (138.03 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 | 1.4135 mL | 7.0677 mL | 14.1355 mL | |
| 5 mM | 0.2827 mL | 1.4135 mL | 2.8271 mL | |
| 10 mM | 0.1414 mL | 0.7068 mL | 1.4135 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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