| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Human Endogenous Metabolite; second messenger; STING/stimulator of interferon genes
cGAMP diammonium directly activates the stimulator of interferon genes (STING), an endoplasmic reticulum (ER)-resident receptor. Activation of STING initiates a signaling cascade via TBK1 and IRF3, leading to the production of type I interferons (IFN-alpha/beta) and other pro-inflammatory cytokines. |
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| ln Vitro |
The ability of mouse splenocytes to proliferate in response to specific antigens is enhanced by cGAMP diammonium [2]. In vitro, human and mouse dendritic cells are immediately activated by cGAMP diammonium [2]. Patient fibroblasts exhibited enhanced transcription of IFNB1 upon stimulation with cGAMP diammonium, but not of genes encoding TNF, IL6, or interleukin 1 (IL1) [3]. The endoplasmic reticulum (ER)-resident receptor STING is activated by cGAMP diammonium, which results in the induction of an antiviral state and type I interferon production [4].
In vitro, cGAMP diammonium directly activates murine and human dendritic cells, promoting their maturation and antigen presentation capabilities. It also stimulates fibroblasts from patients, leading to increased transcription of IFNB1. It induces an antiviral state and the secretion of type I IFNs. |
| ln Vivo |
In mice that have received vaccinations, the spleen cells produce more antigen-specific cytokines when exposed to 5 μg of nasal mucosal adjuvant diammonium (cGAMP) [2].
The recently discovered mammalian enzyme cyclic GMP-AMP synthase produces cyclic GMP-AMP (cGAMP) after being activated by pathogen-derived cytosolic double stranded DNA. The product can stimulate STING-dependent interferon type I signaling. Here, we explore the efficacy of cGAMP as a mucosal adjuvant in mice. In this study, researchers show that cGAMP can enhance the adaptive immune response to the model antigen ovalbumin. It promotes antigen specific IgG and a balanced Th1/Th2 lymphocyte response in immunized mice. A characteristic of the cGAMP-induced immune response is the slightly reduced induction of interleukin-17 as a hallmark of Th17 activity – a distinct feature that is not observed with other cyclic di-nucleotide adjuvants. We further characterize the innate immune stimulation activity in vitro on murine bone marrow-derived dendritic cells and human dendritic cells. The observed results suggest the consideration of cGAMP as a candidate mucosal adjuvant for human vaccines[2]. In vivo, cGAMP diammonium is used as a vaccine adjuvant, where it promotes higher antibody titers when administered intranasally. It activates the cGAS-STING pathway, facilitating innate immune responses and can reinforce antitumor immune responses in animal models of cancer. |
| Enzyme Assay |
Cytosolic DNA induces type I interferons and other cytokines that are important for antimicrobial defense but can also result in autoimmunity. This DNA signaling pathway requires the adaptor protein STING and the transcription factor IRF3, but the mechanism of DNA sensing is unclear. We found that mammalian cytosolic extracts synthesized cyclic guanosine monophosphate-adenosine monophosphate (cyclic GMP-AMP, or cGAMP) in vitro from adenosine triphosphate and guanosine triphosphate in the presence of DNA but not RNA. DNA transfection or DNA virus infection of mammalian cells also triggered cGAMP production. cGAMP bound to STING, leading to the activation of IRF3 and induction of interferon-β. Thus, cGAMP functions as an endogenous second messenger in metazoans and triggers interferon production in response to cytosolic DNA[1].
For in vitro enzyme-binding assays, the affinity of cGAMP for STING is determined using surface plasmon resonance (SPR) or isothermal titration calorimetry (ITC). Purified STING protein is immobilized, and varying concentrations of cGAMP are flowed over the chip to calculate the binding constant (Kd). |
| Cell Assay |
In vitro stimulation of primary cells[2]
The culture medium of primary cells was supplemented with 5 µg/ml (murine cells) or 60 µg/ml (human cells) of c-di-AMP or cGAMP or left without additive. Cells were incubated for 24 h at 37°C. Scrape loading[3] HEK STING cells were seeded at a density of 2.5 × 105 cells ml−1 in 96-well plates. After 16 h cGAMP(2′-5′) was added to the medium to a final concentration of 50 μg ml−1. Monolayers of cells were manually wounded by six scratches per well using an 18G needle. Images were acquired after 4–8 h. In cell-based assays, cells such as THP-1-derived macrophages or HEK293T cells stably expressing a STING-dependent luciferase reporter are plated in 96-well plates. Cells are treated with cGAMP diammonium (e.g., 0.1-100 microg/mL) for 6-24 hours. After incubation, cell lysates are prepared, and luciferase activity is measured. |
| Animal Protocol |
Animal/Disease Models: Female C57BL/6 (H-2b) mice 6-8 weeks old[2]
Doses: 5 µg Route of Administration: Nostril mucosal adjuvant Experimental Results: Higher titers of ovalbumin (OVA)-specific IgA and total IgG as well as IgG1 and IgG2c in the sera of mice immunized with cGAMP-adjuvanted OVA as compared to sera from OVA-immunized mice. Mouse immunization experiments[2] Five animals per group were immunized i. n. on days 0, 14 and 28. Animals were anesthetized with Isoflurane and treated 10 µl per nostril with 15 µg OVA alone or co-administered with 5 µg per dose of c-di-AMP, cGAMP or cholera toxin B subunit in Ampuwa or with Ampuwa alone in the control group (mock immunization). On day 42 after immunization animals were sacrificed and samples were collected. In animal studies, female C57BL/6 mice (6-8 weeks old) are administered cGAMP diammonium via intranasal, intratumoral, or intraperitoneal routes (e.g., 5 microg/mouse). Blood and tissue samples are collected at various time points post-administration. For tumor studies, tumor volume is measured every 2-3 days for 2-3 weeks. |
| ADME/Pharmacokinetics |
Following intraperitoneal administration, cGAMP diammonium exhibits a rapid distribution phase with detectable levels in plasma and lymphoid tissues. It has a relatively short elimination half-life. Its diammonium salt form is water-soluble, which aids in parenteral administration and systemic bioavailability.
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| Toxicity/Toxicokinetics |
cGAMP diammonium is generally considered safe for research use at standard doses. In animal studies, no significant changes in survival rate or overt signs of toxicity have been reported. As it activates immune pathways, careful monitoring for excessive inflammation or cytokine release syndrome is recommended.
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| References | |
| Additional Infomation |
cGAMP diammonium is a water-soluble, synthetic cyclic dinucleotide (cGAMP) that functions as an endogenous second messenger activating the STING pathway, leading to type I interferon production. It is widely used in vaccine adjuvant development, cancer immunotherapy research, and studies of antiviral immunity. This product is for research use only.
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| Molecular Formula |
C20H30N12O13P2
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|---|---|
| Molecular Weight |
708.47
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| Related CAS # |
cGAMP disodium;2407516-83-8;cGAMP;849214-04-6
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| Appearance |
Typically exists as solid at room temperature
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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 :~100 mg/mL (~141.15 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: 33.33 mg/mL (47.05 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 | 1.4115 mL | 7.0575 mL | 14.1149 mL | |
| 5 mM | 0.2823 mL | 1.4115 mL | 2.8230 mL | |
| 10 mM | 0.1411 mL | 0.7057 mL | 1.4115 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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