2'3'-c-di-AM(PS)2 (Rp,Rp) VacciGrade™

2’3’-c-diAM(PS)2(Rp,Rp) endotoxin-free, sterile STING ligand

2’3’-c-di-AM(PS)₂ (Rp,Rp) is the Rp, Rp-isomer of the 2’3’ bisphosphorothioate analog of 3’3’-cyclic adenosine monophosphate (c-di-AMP), a  cyclic dinucleotide (CDN). c-di-AMP is a second messenger molecule produced by bacteria and has potent immunostimulant activity in mammals [1]. It activates innate immunity by directly binding the endoplasmic reticulum-resident receptor STING (stimulator of interferon genes), leading to the expression of interferon-β (IFN-β) and nuclear factor-κB (NF-κB) dependent inflammatory cytokines [2].
 

Key Features

• 2’3’-c-di-AM(PS)₂ (Rp,Rp) has a higher affinity for STING than c-di-AMP due to the presence of a 2’-5’, 3’-5’ mixed linkage, as found in endogenous human CDNs produced by cGAS (cyclic GMP-AMP (cGAMP) synthase) [3]. It activates all known human STING alleles, as well as mouse STING.
   
• In addition, this c-di-AMP analog contains two phosphorothioate diester linkages, which prevent its degradation by the phosphodiesterases that are present in host cells or in the systemic circulation [4].
   
• Finally, this Rp, Rp dithio diastereoisomer was found to induce higher type I IFN production when compared to the Rp/Sp dithio diastereoisomers or c-di-AMP [3].

2’3’-c-di-AM(PS)₂ (Rp,Rp) VacciGrade™ is a high-quality pre-clinical grade.
2’3’-c-di-AM(PS)₂ (Rp,Rp)​ VacciGrade™​ is for research use only, and not for human or veterinary use.

 Read our review on STING: Deciphering the STING Paradox

References:

1. Woodward JJ. et al., 2010. c-di-AMP secreted by intracellular Listeria monocytogenes activates a host type I interferon response. Science.328(5986):1703-5.
2. Jin L. et al., 2011. MPYS is required for IFN response factor 3 activation and type I IFN production in the response of cultured phagocytes to bacterial second messengers cyclic-di-AMP and cyclic-di- GMP. J Immunol. 187(5):2595-601.
3. Corrales L et al., 2015. Direct activation of STING in the tumor microenvironment leads to potent and systemic tumor regression and immunity. Cell Rep. 11(7):1018-30.
4. Yan H. et al., 2008. Synthesis and immunostimulatory properties of the phosphorothioate analogs of cdiGMP. Bioorg. Med. Chem. Lett. 18, 5631–5634.

Figures

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). The interferon regulatory factor (IRF) response was assessed by measuring the activity of Lucia luciferase in the supernatant using QUANTI-Luc™ 4 Lucia/Gaussia. Data are shown in fold response over non-induced cells (mean ± SEM).

THP1-Dual™ cells were stimulated for 24 hours with the STING ligands as shown (all at 10 μg/ml). The NF-κB-induced SEAP activity was assessed using QUANTI‑Blue™ Solution, a SEAP detection reagent. Data are shown as optical density (OD) at 630 nm (mean ± SEM). Non‑induced cells (NI) have been included as a negative control.

Specifications

Description: STING agonist VacciGrade™

Synonyms: (R,R)-(2',3')c-diAM(PS)₂, (2',3')-Rp,Rp-c-diAMPSS

CAS number: 1638750-95-4 / 1638241-89-0 (free acid)

Formula: C₂₀H₂₂N₁₀O₁₀P₂S₂•2Na

Molecular weight: 734.50 g/mol

Purity: ≥ 95% by LC/MS and NMR

Solubility: 50 mg/ml in water

Quality control:

• Sterility guaranteed
• The absence of bacterial contamination (lipoproteins & endotoxins) has been confirmed using HEK-Blue™ TLR2 and HEK-Blue™ TLR4 cells
• Endotoxin level < 5 EU/mg (determined using the HEK-Blue™ LPS Detection Kit 2)
• The ability of 2’3’-c-di-AM(PS)₂ (Rp,Rp) to induce type I interferon (IFN) has been confirmed in cellular assays.

Contents

• 500 μg of lyophilized 2’3’-c-di-AM(PS)₂ (Rp,Rp) VacciGrade™
• 10 ml sterile endotoxin-free physiological water (NaCl 0.9%)

2'3'-c-di-AM(PS)₂ (Rp,Rp) VacciGrade™ is shipped at room temperature and

2'3'-c-di-AM(PS)₂ (Rp,Rp) VacciGrade™ should be stored at -20°C.

VacciGrade™

VacciGrade™ is a high-quality pre-clinical grade. VacciGrade™ products are filter-sterilized (0.2 µm) and filled under strict aseptic conditions in a clean room*. The absence of bacterial contamination is assessed by a sterility test using a pharmacopeia-derived assay. The level of bacterial contaminants (endotoxins and lipoproteins) in each lot is verified using a LAL assay and/or a TLR2 and TLR4 reporter assay.
*Except for LPS VacciGrade™, which is prepared in a laminar flow hood dedicated to LPS.

Details