| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg | |||
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| Other Sizes |
| Targets |
STING/stimulator of interferon genes
The target of ADU-S100 disodium salt is STING (stimulator of interferon genes), a critical component of the innate immune sensing pathway. The compound binds to and activates STING, triggering downstream signaling through TBK1 and IRF3 phosphorylation, which leads to type I interferon production and activation of immune cells. ADU-S100 activates human STING and human STING variants including STING HAQ, REF, AQ, and Q, as well as mouse STING. |
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| ln Vitro |
In THP-1 human monocytes, ADU-S100 produced more type I IFN than CDA. Disulfide mixed-linked cyclic dinucleotide (CDN) derivatives, on the other hand, effectively activated all five hSTING alleles, including the refractory hSTINGREF and hSTINGQ alleles (ML RR-CDA, ML RR-S2 CDG, and ML RR-S2 cGAMP). In comparison to endogenous ML cGAMP and the TLR3 agonist Poly I:C, ADU-S100 induced the highest expression of IFN-β and the proinflammatory cytokines TNF-α, IL-6, and MCP-1 on a molar equivalent basis. In mouse bone marrow macrophages (BMM), ADU-S100 was also found to induce STING aggregation and induce TBK1 and IRF3 phosphorylation. When compared to ML cGAMP, ADU-S100 induces noticeably higher levels of IFN-α [1].
ADU-S100 disodium salt demonstrates potent in vitro activity as a STING agonist. It shows enhanced type I IFN production compared to CDA in THP-1 human monocytes. The compound activates STING signaling in various cell types, leading to interferon production and immune activation. At 10 microM concentration, it induces IFN-beta-dependent reporter activity in HEK293T cells expressing human or mouse STING. The disodium salt form provides favorable solubility properties. |
| ln Vivo |
ADU-S100 outperformed endogenous ML cGAMP in its anti-tumor control capacity. In B16 tumor-bearing mice, a dose response study of ADU-S100 compounds was carried out to ascertain the best anti-tumor dose level that would maximize tumor antigen-specific CD8+ T cell responses and increase long-term survival to 50%[1].
In vivo, ADU-S100 disodium salt leads to potent anti-tumor activity through STING-mediated immune activation. The compound induces systemic immune responses that promote tumor regression. It effectively activates the STING pathway in vivo, resulting in robust anti-tumor immunity. The compound has been evaluated in various preclinical tumor models. Systemic administration induces both local and systemic anti-tumor immune responses. |
| Enzyme Assay |
Luciferase Assay[1]
104 HEK293T cells were seeded in 96-well plates and transiently transfected with human IFN-β firefly reporter plasmid(Fitzgerald et al., 2003) and TK-Renilla luciferase reporter for normalization. The following day, cells were stimulated with 10 μM of eachADU-S100 or 100 μg/ml DMXAA using digitonin permeabilization (50 mM HEPES, 100 mM KCL, 3 mM MgCl2, 0.1 mM DTT, 85 mM Sucrose, 0.2% BSA, 1 mM ATP, 0.1 mM GTP, 10 ug/ml digitonin) to ensure uniform uptake. After 20 min, stimulation mixtures were removed and normal media was added. After a total of 6 hours, cell lysates were prepared and reporter gene activity measured using the Dual Luciferase Assay System on a Spectramax M3 luminometer. Differential Scanning Fluorimetry[1] Thermal shift assays were performed as (Cavlar et al., 2013). Assays were conducted with STING ligand binding domain at 1 mg/ml with or without various ADU-S100 at 1 mM in 20mM Tris-HCL, 150 mM NaCl, pH 7.5 and 1:500 dilution of SYPRO Orange Dye. The fluorescence as a function of temperature was recorded in a CFX 96 real time PCR machine reading on the HEX channel EX 450–490 EM 560–580 nm. The temperature gradient was from 15–80°C ramping 0.5°C per 15 seconds. Curves were fit to a Boltzmann sigmoidal to establish the midpoint of thermal unfolding (Tm). In vitro enzyme/receptor binding assays for ADU-S100 disodium salt involve binding affinity studies to STING protein using techniques such as surface plasmon resonance (SPR) or fluorescence polarization. STING activation is assessed using reporter cell lines expressing STING and interferon-sensitive reporter genes. Cells are treated with varying concentrations of ADU-S100 (typically 0.001-100 microM range), and reporter activity is measured after 4-24 hours. IC50 values are determined from dose-response curves. |
| Cell Assay |
BM-DCs from WT or STING−/− mice were stimulated with 25 μg/ml DMXAA or 100 ng/ml LPS for 4 hours. Total RNA was isolated using the RNeasy® kit and incubated with Deoxyribonuclease I, Amplification Grade. cDNA was synthesized using High Capacity cDNA Reverse Transcription Kit and expression of cytokines was measured by real-time qRT-PCR using specific primers/probes for mouse INF-β, TNF-α, IL-6 and IL12p40, and pan-specific primers were to quantify expression of the IFN-α family. Primer sequences are listed in Table 1 in Supplementary Materials. PCR reactions were performed in the 7300 Real Time PCR system. The results are expressed as 2−ΔCt using 18s as endogenous control.
WT BMM were stimulated with ADU-S100 at 5 μM in HBSS with the addition of Effectene transfection reagent (per kit protocol). Human PBMCs were stimulated as indicated. Stimulated cells were and assessed by real-time qRT-PCR for gene expression of IFN-β1, MCP-1, TNF-α and IL-6 using the PrimePCR RNA purification and cDNA analysis system, and run on the CFX96 gene cycler. Relative normalized expression was determined by comparing induced target gene expression to unstimulated controls, using the reference genes Gapdh and Ywhaz (mouse) and GusB and Pgk1 (human), genes confirmed to have a coefficient variable (CV) below 0.5 and M value below 1, and thus did not vary with different treatment conditions. In vitro cellular assays for ADU-S100 disodium salt involve treating STING-expressing cell lines with varying concentrations (typically 0.001-100 microM range) for 4-24 hours. Readouts include type I interferon production measured by ELISA, qPCR for interferon-stimulated genes, and phosphorylation of pathway components (TBK1, IRF3) by Western blotting. HEK293T cells expressing human or mouse STING are commonly used for reporter assays. THP-1 monocytes are used for IFN production studies. |
| Animal Protocol |
10~6 of B16-SIY tumor cells, 5 × 10~4 B16.F10 tumor cells, 10~5 4T-1 and CT26, or 106 other tumor cells were injected s.c. in 100 μl DPBS or HBSS on the right flank of mice. Following tumor implantation, mice were randomized into treatment groups. When tumors were 100–200 mm3 in volume (5–7 mm wide), either one single or three doses of DMXAA resuspended in 7.5% of NaHCO3, or CDNs formulated in HBSS or vehicle control, were injected IT. Measurements of tumors were performed twice per week using calipers, and the tumor volume was calculated with the formula: V= (length × width2)/2. In some experiments, tumor-free survivors were rechallenged with tumor cells on the opposite flank several weeks after the injection of the primary tumor. Naïve mice were used as controls. For the contralateral experiments, mice were implanted on both flanks and only one tumor was treated. For the B16 melanoma lung metastasis experiments, mice were implanted on the flank with 5 × 104 cells B16.F10 on day 0, and then injected intravenously with 1 × 105 cells on day 7. Lungs were harvested on day 28. Administration of compounds, measurements of tumors and counting of lung tumors were performed in a blinded fashion.
In vivo animal studies for ADU-S100 disodium salt typically employ syngeneic tumor models in immunocompetent mice. Tumor-bearing mice are administered the compound via various routes including intratumoral or systemic injection at doses typically ranging from 0.1-10 mg/kg. Tumor growth inhibition is monitored over 2-4 weeks. Endpoints include tumor volume, immune cell infiltration, cytokine profiling, and survival. Rechallenge studies assess establishment of immune memory. |
| ADME/Pharmacokinetics |
Pharmacokinetic properties of ADU-S100 disodium salt are characterized by typical cyclic dinucleotide PK parameters. Following administration, the compound shows appropriate biodistribution. PK parameters including half-life, clearance, and volume of distribution are determined via LC-MS/MS analysis. The disodium salt form provides favorable solubility and stability properties. Bioavailability depends on the route of administration. The compound is typically formulated in appropriate buffers for in vivo studies.
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| Toxicity/Toxicokinetics |
Toxicological data for ADU-S100 disodium salt indicates that it is a research-grade compound. As a STING agonist, potential toxicities are related to immune activation and cytokine release. The compound has been evaluated in preclinical safety studies. Standard laboratory safety precautions should be followed during handling. Comprehensive toxicology data would be required for clinical development. The compound is for research use only and not for therapeutic purposes.
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| References | |
| Additional Infomation |
ADU-S100 (MIW815) is a synthetic cyclic dinucleotide (CDN) agonist (activator) that activates the interferon gene-stimulating factor (STING) receptor. The STING receptor is crucial for activating the innate (endogenous) immune system. ADU-S100 (MIW815) activates all known human and mouse STING receptors and effectively induces the expression of cytokines and chemokines, thereby generating a potent and durable antigen-specific T cell-mediated immune response against cancer cells. The STING-activating cyclic dinucleotide agonist MIW815 is a synthetic cyclic dinucleotide (CDN) and an agonist of the interferon gene-stimulating factor protein (STING; transmembrane protein 173; TMEM173), possessing potential immunomodulatory and antitumor activity. Upon intratumoral injection, the STING agonist MIW815 binds to STING and activates the STING-mediated signaling pathway. This activates the immune response by activating certain immune cells, including dendritic cells (DCs), which in turn induces the expression of cytokines and chemokines, ultimately leading to an antigen-specific T cell-mediated immune response against cancer cells. STING is a transmembrane protein that can activate immune cells in the tumor microenvironment and plays a key role in the activation of the innate immune system.
ADU-S100 disodium salt (CAS# 1638750-95-4) is also known as MIW815 and ML RR-S2 CDA. It is a STING agonist that has been investigated in clinical trials for cancer immunotherapy. The compound activates the innate immune system to promote anti-tumor immunity. ADU-S100 has been studied in combination with immune checkpoint inhibitors. The disodium salt form offers improved solubility and pharmaceutical properties. The compound is typically stored at -20degC. |
| Molecular Formula |
C20H22N10NA2O10P2S2
|
|---|---|
| Molecular Weight |
734.506742954254
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| Exact Mass |
734.023
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| Elemental Analysis |
C, 32.71; H, 3.02; N, 19.07; Na, 6.26; O, 21.78; P, 8.43; S, 8.73
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| CAS # |
1638750-95-4
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| Related CAS # |
ADU-S100 ammonium salt;1638750-96-5;ADU-S100 enantiomer ammonium salt;ADU-S100;1638241-89-0
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| PubChem CID |
135390762
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| 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 |
1180
|
| Defined Atom Stereocenter Count |
8
|
| SMILES |
S=P1([O-])OC[C@@H]2[C@H]([C@H]([C@H](N3C=NC4C(N)=NC=NC3=4)O2)O)OP([O-])(OC[C@@H]2[C@H]([C@H]([C@H](N3C=NC4C(N)=NC=NC3=4)O2)O1)O)=S.[Na+].[Na+]
|
| InChi Key |
GDWOOOCBNOMMTL-ITQXCAEYSA-L
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| InChi Code |
InChI=1S/C20H24N10O10P2S2.2Na/c21-15-9-17(25-3-23-15)29(5-27-9)19-12(32)13-8(38-19)2-36-42(34,44)40-14-11(31)7(1-35-41(33,43)39-13)37-20(14)30-6-28-10-16(22)24-4-26-18(10)30/h3-8,11-14,19-20,31-32H,1-2H2,(H,33,43)(H,34,44)(H2,21,23,25)(H2,22,24,26)/q2*+1/p-2/t7-,8-,11-,12-,13-,14-,19-,20-,41-,42-/m1../s1
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| Chemical Name |
(2',3')-Rp,Rpc-diAMPS disodium
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| Synonyms |
ML RR-S2 CDA disodium salt; ADU-S100; MIW815 disodium salt; ADU-S100; Cdn agonist adu-S100; 1638750-95-4; FMW9ZVF53N; ML RR-S2 CDA Sodium Salt; ADU S100 [WHO-DD]; (2',3')-Rp,Rpc-diAMPS disodium; MW-815; ADU S100; MIW 815 disodium salt; ADUS100; MIW-815 disodium salt; 2’3’-c-di-AM(PS)2(Rp,Rp)Bisphosphorothioate analog of c-di-AMP, Rp isomers
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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 (~136.15 mM)
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|---|---|
| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 25 mg/mL (34.04 mM) (saturation unknown) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution.
(Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.3615 mL | 6.8073 mL | 13.6145 mL | |
| 5 mM | 0.2723 mL | 1.3615 mL | 2.7229 mL | |
| 10 mM | 0.1361 mL | 0.6807 mL | 1.3615 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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