cAIMP Difluor (CL614)

cAIMP difluorinated

InvivoGen's cAIMP Difluor (also known as CL614) is a derivative of cAIMP, an analog of the bacterial cyclic dinucleotide (CDN) 3’3’-cGAMP [1].

cAIMP and cAIMP Difluor are novel STING (stimulator of interferon genes)-activating synthetic CDNs.

Unlike natural CDNs, whose constituent nucleosides are guanosine and/or adenine, cAIMP and its derivatives contain one adenine nucleoside and one inosine nucleoside.

cAIMP Difluor is composed of two 2’-deoxynucleosides with a fluorine atom at 2’ position of each nucleoside.

STING ligands such as cAIMP induce the production of type I interferons (IFNs) and of proinflammatory cytokines through the IRF and NF-κB pathways, respectively.

The incorporation of fluorine into biologically active molecules is commonly used in medicinal chemistry to improve their metabolic stability or to modulate physicochemical properties such as lipophilicity [2]. Moreover, the introduction of a fluorine atom can change the biological activities of a molecule. Indeed cAIMP

Difluor (referred to as compound 52 by Lioux et al.  [1]) more potently induces interferon regulatory factor (IRF) and NF-κB pathways in a STING‑dependent manner when compared to STING agonists such as 2’3’‑cGAMP and DMXAA [1]. Interestingly, cAIMP Difluor is more resistant than 2’3’‑cGAMP and cAIMP to cleavage by certain nuclease and phosphodiesterase enzymes [1].

To facilitate the srudy of STING ligands, InvivoGen has developed stable reporter cells in two well established immune cell models: THP-1 human monocytes and RAW 264.7 murine macrophages. These cells express inducible SEAP and/or Lucia luciferase reporter genes under the control of an IRF-inducible and NF-κB promoter. 

References:

1. Lioux T. et al., 2016. Design, synthesis, and biological evaluation of novel cyclic adenosine-inosine monophosphate (cAIMP) analogs that activate stimulator of interferon genes (STING). J Med Chem. 59(22):10253-10267.
2. Böhm HJ. et al., 2004. Fluorine in medicinal chemistry. Chembiochem. 5(5):637-43

Figures

THP1-Dual™ cells were stimulated for 24 hours with human IFN-β (1 x 10⁴ U/ml), TNF-α (300 pg/ml), cAIMP, cAIMP Difluor, cAIM(PS)2 Difluor (Rp/Sp), 2’3’-cGAMP and 2’3’-cGAM(PS)2 (Rp/Sp). All CDNs were used at 10 μg/ml.
IRF induction was determined by measuring the relative light units (RLUs) in a luminometer using QUANTI‑Luc™, a Lucia luciferase detection reagent. The IRF induction of each ligand is expressed relative to that of hIFN-β at 1 x 10⁴ U/ml (taken as 100%).

THP1-Dual™ cells were stimulated for 24 hours with human IFN-β (1x10⁴U/ml), TNF-α (300pg/ml), cAIMP, cAIMP Difluor, cAIM(PS)2 Difluor (Rp/Sp), 2’3’-cGAMP and 2’3’-cGAM(PS)2 (Rp/Sp). All CDNs were used at 10μg/ml.
NF-κB induction was determined using QUANTI‑Blue™, a SEAP detection reagent, and by reading the optical density (OD) at 655 nm. TNF-α has been included as a positive control to activate the NF-κB signaling pathway.

Specifications

Source: Synthetic

Synonym: c-(2’FdAMP-2’FdIMP) sodium salt

CAS: 1951464-78-0

Formula: C₂₀H₂₁F₂N₉O₁₁P₂ .2Na

Molecular weight: 663.4 (free acid), 707.4 (sodium salt)

Solubility: 50 mg/ml in water

Quality control:

• Purity and structure has been determined by LC/MS and NMR: ≥ 95%
• The biological activity of cAIMP Difluor has been confirmed using cellular assays.
• The absence of bacterial contamination (e.g. lipoproteins & endotoxins) has been confirmed using HEK-Blue™ TLR4 and HEK-Blue™ TLR2 cells.

Contents

• 2 x 250 μg of cAIMP Difluor provided lyophilized. 
  Note: cAIMP Difluor is sterile filtered prior to lyophilization.
• 1.5 ml endotoxin-free water

Product is shipped at room temperature

 Upon receipt, store at -20 °C.

Description

cAIMP Difluor chemical structure