2',3'-cGAMP sodium
(Synonyms: 2'-3'-cyclic GMP-AMP sodium) 目录号 : GC39324
2',3'-cGAMP 钠（2'-3'-环状 GMP-AMP 钠）是哺乳动物细胞中的内源性 cGAMP。
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
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- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
Bone marrow cells isolated from WT mice and STINGgt mice |
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Preparation Method |
Bone marrow cells were isolated from WT mice and STINGgt mice and differentiated into macrophages (BMDM)，After differentiation, both WT BMDM and STINGgt BMDM were treated with or without 2’,3’-cGAMP sodium (20 µg/ml) for 24h in the presence or absence of lipopolysaccharide (LPS) (20 ng/ml) for the last 6 hr. In parallel, both WT BMDM and STINGgt BMDM were treated with commercial 2’,3’-cGAMP sodium |
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Reaction Conditions |
Cells treated at a dose of 20 µg/ml for 0, 6, 24, and/or 48h or at a dose of 0, 5, 20 and/or 40 µg/ml for 24h. |
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Applications |
WT BMDM treated with enzymatically synthesized 2’,3’-cGAMP sodium and observed significant increases in LPS-stimulated proinflammatory signaling through JNK p46 and NF-κB p65 and expression of IL-1β, IL-6, and TNFα. |
| Animal experiment [2]: | |
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Animal models |
Female nude Foxn1 mice 6-week-old |
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Preparation Method |
B16F10 melanoma tumor cells were injected into the right shoulder of female mice to establish tumors. At 7 days after tumor cell inoculation, the mice were treated with zebularine alone, 2’,3’-cGAMP sodium alone, or the combination of both. A final concentration of 2.5 mg/mL zebularine was added to drinking water for 10 days until the mice were collected for conducting further experiments. A total of 100 μL of 10μg 2’,3’-cGAMP sodium in PBS at the indicated concentrations was injected into the site next to tumor. 2’,3’-cGAMP sodium treatment was repeated three times with 4-day intervals. |
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Dosage form |
10μg, three times every the 4th day, injected into the site next to tumor |
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Applications |
At this time point, we saw a similar effect from triple combination compared to the 2’,3’-cGAMP sodium +zebularine group. However, further statistics on the tumor growth and survival curve showed that triple combination led to more robust control of tumor growth and markedly extended survival in the B16F10 model. |
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References: [1]. X. Guo, C. Shu, H. Li, et al. Cyclic GMP-AMP ameliorates diet-induced metabolic dysregulation and regulates proinflammatory responses distinctly from STING activation Sci Rep, 7 (2017), p. 6355 [2]. Lai J, Fu Y, Tian S, Huang S, Luo X, Lin L, Zhang X, Wang H, Lin Z, Zhao H et al (2021) Zebularine elevates STING expression and enhances cGAMP cancer immunotherapy in mice. Mol Ther 29: 1758–1771 |
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2′3′-cGAMP sodium is a second messenger that binds and activates the adaptor protein stimulator of interferon (STING), which triggers the innate immune response [1]. As a STING agonist, the small molecule 2′3′-cGAMP sodium plays pivotal roles in antiviral defense and has adjuvant applications, and anti-tumor effects. 2′3′-cGAMP sodium and its analogs are thus putative targets for immunotherapy and are currently being tested in clinical trials to treat solid tumors.
2′3′-cGAMP sodium is capable of enhancing the proinflammatory activation of cultured Wild-type (WT) macrophages. Unlike in macrophages (BMDM), 2′3′-cGAMP sodium treatment displayed anti-inflammatory effects in both WT primary mouse hepatocytes and differentiated 3T3-L1 adipocytes. Specifically, LPS-induced JNK p46 and NF-κB p65 phosphorylation states and IL-1β and TNFα mRNAs in 2′3′-cGAMP sodium -treated WT primary mouse hepatocytes were significantly lower than their respective levels in control-treated hepatocytes. In 3T3-L1 adipocytes, the anti-inflammatory effect of 2′3′-cGAMP sodium was even more pronounced. In particular, LPS-induced JNK p46 phosphorylation states in 2′3′-cGAMP sodium -treated adipocytes were markedly lower than in control-treated adipocytes, and were comparable with JNK p46 phosphorylation states in 2′3′-cGAMP sodium -treated adipocytes in the absence of LPS induction [2].
2′3′-cGAMP sodium and CpG-C co-administration adjuvants had a synergistic effect to establish a shift towards the Th1(T helper type 1) immune response, and leading to reduced tumor growth. This vaccine formulation could be a promising therapeutic candidate vaccine for HPV 16 established infections and HPV-associated tumors [3]. 2′3′-cGAMP sodium led to a marked CD8 T cell increase in tumors; combined treatment further increased the percentage of CD8 T cells [4].
References:
[1]. Su M, Zheng J, Gan L, Zhao Y, Fu Y, Chen Q. Second Messenger 2’3’-Cyclic GMP-AMP (2’3’-cGAMP):Synthesis, Transmission, and Degradation. Biochem Pharmacol (2022) 198:114934. doi: 10.1016/j.bcp.2022.114934
[2]. X. Guo, C. Shu, H. Li, et al. Cyclic GMP-AMP ameliorates diet-induced metabolic dysregulation and regulates proinflammatory responses distinctly from STING activation Sci Rep, 7 (2017), p. 6355
[3]. Dorostkar, F.; Arashkia, A.; Roohvand, F. et al. Co-administration of 2’3’-cGAMP STING activator and CpG-C adjuvants with a mutated form of HPV 16 E7 protein leads to tumor growth inhibition in the mouse model. Infect. Agents Cancer 2021, 16, 7.
[4]. Lai J, Fu Y, Tian S, et al. Zebularine elevates STING expression and enhances cGAMP cancer immunotherapy in mice. 2021. Mol Ther 29: 1758–1771.
2'3'-cGAMP 钠是第二信使，可结合并激活干扰素 (STING) 的衔接蛋白刺激因子，从而触发先天免疫反应[1]。作为 STING 激动剂，小分子 2'3'-cGAMP 钠在抗病毒防御中起着关键作用，并具有辅助应用和抗肿瘤作用。 2'3'-cGAMP 钠及其类似物因此被推定为免疫治疗的靶标，目前正在临床试验中测试以治疗实体瘤。
2'3'-cGAMP 钠能够增强培养的野生型 (WT) 巨噬细胞的促炎激活。与巨噬细胞 (BMDM) 不同，2'3'-cGAMP 钠处理在 WT 原代小鼠肝细胞和分化的 3T3-L1 脂肪细胞中均显示出抗炎作用。具体而言，LPS 诱导的 JNK p46 和 NF-κB p65 磷酸化状态以及 2'3'-cGAMP 钠处理的 WT 原代小鼠肝细胞中的 IL-1β 和 TNFα mRNA 显着低于其在对照处理的肝细胞中各自的水平。在 3T3-L1 脂肪细胞中，2'3'-cGAMP 钠的抗炎作用更为明显。特别是，在 2'3'-cGAMP 钠处理的脂肪细胞中，LPS 诱导的 JNK p46 磷酸化状态明显低于对照处理的脂肪细胞，并且与 2'3'-cGAMP 钠处理的脂肪细胞中的 JNK p46 磷酸化状态相当在没有 LPS 诱导的情况下 [2]。
2'3'-cGAMP 钠和 CpG-C 联合给药佐剂具有协同作用，可建立向 Th1（T 辅助细胞 1 型）免疫反应的转变，并导致肿瘤生长减少。这种疫苗制剂可能是一种很有前途的治疗性候选疫苗，可用于治疗 HPV 16 感染和 HPV 相关肿瘤[3]。 2'3'-cGAMP 钠导致肿瘤中 CD8 T 细胞显着增加；联合治疗进一步提高了CD8 T细胞的比例[4]。
| Cas No. | SDF | ||
| 别名 | 2'-3'-cyclic GMP-AMP sodium | ||
| Canonical SMILES | OC1([H])[C@](OP(OC[C@](O[C@@H](N2C3=NC=NC(N)=C3N=C2)[C@@H]4O)([H])[C@@]4([H])O5)(O[Na])=O)([H])[C@H](N6C(NC(N)=NC7=O)=C7N=C6)O[C@]1([H])COP5(O[Na])=O | ||
| 分子式 | C20H22N10Na2O13P2 | 分子量 | 718.37 |
| 溶解度 | Water: 50 mg/mL (69.60 mM); DMSO: 12.5 mg/mL (17.40 mM) | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.392 mL | 6.9602 mL | 13.9204 mL |
| 5 mM | 0.2784 mL | 1.392 mL | 2.7841 mL |
| 10 mM | 0.1392 mL | 0.696 mL | 1.392 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
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cGAS-like receptors sense RNA and control 3'2'-cGAMP signalling in Drosophila
Nature2021 Sep;597(7874):109-113.PMID: 34261127DOI: 10.1038/s41586-021-03743-5
Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor that produces the second messenger cG[2'-5']pA[3'-5']p (2'3'-cGAMP) and controls activation of innate immunity in mammalian cells1-5. Animal genomes typically encode multiple proteins with predicted homology to cGAS6-10, but the function of these uncharacterized enzymes is unknown. Here we show that cGAS-like receptors (cGLRs) are innate immune sensors that are capable of recognizing divergent molecular patterns and catalysing synthesis of distinct nucleotide second messenger signals. Crystal structures of human and insect cGLRs reveal a nucleotidyltransferase signalling core shared with cGAS and a diversified primary ligand-binding surface modified with notable insertions and deletions. We demonstrate that surface remodelling of cGLRs enables altered ligand specificity and used a forward biochemical screen to identify cGLR1 as a double-stranded RNA sensor in the model organism Drosophila melanogaster. We show that RNA recognition activates Drosophila cGLR1 to synthesize the novel product cG[3'-5']pA[2'-5']p (3'2'-cGAMP). A crystal structure of Drosophila stimulator of interferon genes (dSTING) in complex with 3'2'-cGAMP explains selective isomer recognition, and 3'2'-cGAMP induces an enhanced antiviral state in vivo that protects from viral infection. Similar to radiation of Toll-like receptors in pathogen immunity, our results establish cGLRs as a diverse family of metazoan pattern recognition receptors.
Hydrolysis of 2'3'-cGAMP by ENPP1 and design of nonhydrolyzable analogs
Nat Chem Biol2014 Dec;10(12):1043-8.PMID: 25344812DOI: 10.1038/nchembio.1661
Agonists of mouse STING (TMEM173) shrink and even cure solid tumors by activating innate immunity; human STING (hSTING) agonists are needed to test this therapeutic hypothesis in humans. The endogenous STING agonist is 2'3'-cGAMP, a second messenger that signals the presence of cytosolic double-stranded DNA. We report activity-guided partial purification and identification of ecto-nucleotide pyrophosphatase/phosphodiesterase (ENPP1) to be the dominant 2'3'-cGAMP hydrolyzing activity in cultured cells. The hydrolysis activity of ENPP1 was confirmed using recombinant protein and was depleted in tissue extracts and plasma from Enpp1(-/-) mice. We synthesized a hydrolysis-resistant bisphosphothioate analog of 2'3'-cGAMP (2'3'-cG(s)A(s)MP) that has similar affinity for hSTING in vitro and is ten times more potent at inducing IFN-β secretion from human THP1 monocytes. Studies in mouse Enpp1(-/-) lung fibroblasts indicate that resistance to hydrolysis contributes substantially to its higher potency. 2'3'-cG(s)A(s)MP is therefore improved over natural 2'3'-cGAMP as a model agonist and has potential as a vaccine adjuvant and cancer therapeutic.
2'3'-cGAMP triggers a STING- and NF-κB-dependent broad antiviral response in Drosophila
Sci Signal2020 Dec 1;13(660):eabc4537.PMID: 33262294DOI: 10.1126/scisignal.abc4537
We previously reported that an ortholog of STING regulates infection by picorna-like viruses in Drosophila In mammals, STING is activated by the cyclic dinucleotide 2'3'-cGAMP produced by cGAS, which acts as a receptor for cytosolic DNA. Here, we showed that injection of flies with 2'3'-cGAMP induced the expression of dSTING-regulated genes. Coinjection of 2'3'-cGAMP with a panel of RNA or DNA viruses resulted in substantially reduced viral replication. This 2'3'-cGAMP-mediated protection was still observed in flies with mutations in Atg7 and AGO2, genes that encode key components of the autophagy and small interfering RNA pathways, respectively. By contrast, this protection was abrogated in flies with mutations in the gene encoding the NF-κB transcription factor Relish. Transcriptomic analysis of 2'3'-cGAMP-injected flies revealed a complex response pattern in which genes were rapidly induced, induced after a delay, or induced in a sustained manner. Our results reveal that dSTING regulates an NF-κB-dependent antiviral program that predates the emergence of interferons in vertebrates.
The Chemistry of the Noncanonical Cyclic Dinucleotide 2'3'-cGAMP and Its Analogs
Handb Exp Pharmacol2017;238:359-384.PMID: 27392950DOI: 10.1007/164_2015_43
The cyclic dinucleotides (CDNs) cyclic diguanosine monophosphate (c-diGMP) and cyclic diadenosine monophosphate (c-diAMP) with two canonical 3'→5' internucleotide linkages are ubiquitous second messenger molecules in bacteria, regulating a multitude of physiological processes. Recently the noncanonical CDN cyclic guanosine monophosphate-adenosine monophosphate (2'3'-cGAMP) featuring a mixed linkage, which consists of a 2'→5' and a 3'→5' internucleotide bond, has been identified as a signaling molecule in metazoan species in late 2012. 2'3'-cGAMP formation is biocatalyzed by cGAMP synthase (cGAS) upon sensing of cytosolic double-stranded DNA (dsDNA) and functions as an endogenous inducer of innate immunity by directly binding to and activating the adaptor protein stimulator of interferon genes (STING). Thereby 2'3'-cGAMP can stimulate interferon-β (INF-β) secretion, a major signaling pathway of host defense, which is independent of toll-like receptor (TLR) activation. Medicinal chemistry of 2'3'-cGAMP and development of corresponding analogs are still in their infancy, and only a handful of structurally related compounds are available to the scientific community. The aim of this chapter is to summarize synthetic approaches to prepare canonical and noncanonical endogenous CDNs including 2'3'-cGAMP. Furthermore, we will describe syntheses of 2'3'-cGAMP analogs bearing modifications, which will facilitate further studies of the emerging biological functions of 2'3'-cGAMP and to identify additional receptor proteins. Finally, we will review latest developments concerning 2'3'-cGAMP analogs with improved hydrolytic stability in cell cultures and in tissues, putatively qualifying for new therapeutic options on the basis of 2'3'-cGAMP signaling.
A Click-Chemistry Linked 2'3'-cGAMP Analogue
Chemistry2019 Feb 6;25(8):2089-2095.PMID: 30536650DOI: 10.1002/chem.201805409
2'3'-cGAMP is an uncanonical cyclic dinucleotide where one A and one G base are connected via a 3'-5' and a unique 2'-5' linkage. The molecule is produced by the cyclase cGAS in response to cytosolic DNA binding. cGAMP activates STING and hence one of the most powerful pathways of innate immunity. cGAMP analogues with uncharged linkages that feature better cellular penetrability are currently highly desired. Here, the synthesis of a cGAMP analogue with one amide and one triazole linkage is reported. The molecule is best prepared via a first CuI -catalyzed click reaction, which establishes the triazole, while the cyclization is achieved by macrolactamization.