CL264 是一种用于先天免疫信号研究的 TLR7 特异性激动剂。
货号:ajcx16484
CAS:1510712-69-2
分子式:C19H23N7O4
分子量：413.43
溶解度:Soluble in DMSO
纯度：98%
存储：Store at -20°C
库存：现货

Background:

CL264 is a a potent and highly specific toll-like receptor 7 (TLR7) ligand. Immune cells utilize toll-like receptors (TLRs) to sense pathogen associated molecular patterns (PAMPs), which represents the starting point of the innate immune response s^[1].

Using concentrations from 0.5 to 10?μg/mL, stimulation with CL264 resulted in a dose-dependent secretion of TNF-α?after 6 hours. CAL-1 cells were stimulated with CL264 (5?μg/mL) for up to 12 hours. ?CAL-1 cells were seeded into 96-well plates. After overnight resting, cells were stimulated with CL264 or 9.2s RNA or the combination of both. After incubation as indicated, cell supernatant was analyzed for TNF-α, IL-6, and IFN-β?by ELISA. CL264 led to a robust, time-dependent release of TNF-α?reaching absolute levels of 1347?pg/mL TNF-α?into the supernatant^[1].Peripheral blood mononuclear cells (PBMCs) stimulated with CL264, IL-6, IL-8 and IL-12 were significantly stimulated^[2].

The TLR7 agonist CL264 ,40 μg/injectionat day 0, 3 and 6 after tumor grafting, stimulated tumor growth, and this pro-tumorigenic effect was completely lost in the absence of MDSCs, both in WT mice and in TLR7 KO mice. The tumor growth exacerbating effect of TLR7 stimulation is mediated by the increased number of MDSCs within the tumor microenvironment.The secretion of CCL2 and GM-CSF by tumor cells was confirmed in vitro, which was found increased in supernatants of CL264-stimulated cells,these results suggest that CL264 may stimulate the recruitment of MDSCs via the induction of CCL2 and GM-CSF.^[3].

参考文献:
[1].Hilbert T, Steinhagen F, et al. Synergistic Stimulation with Different TLR7 Ligands Modulates Gene Expression Patterns in the Human Plasmacytoid Dendritic Cell Line CAL-1. Mediators Inflamm. 2015;2015:948540.
[2].Dajon M, Iribarren K, et al. Toll like receptor 7 expressed by malignant cells promotes tumor progression and metastasis through the recruitment of myeloid derived suppressor cells. Oncoimmunology. 2018 Oct 11;8(1):e1505174.

Protocol:

Cell experiment [1]:
Cell lines	Lewis lung carcinoma (LLC),Bone marrow derived macrophages(BMSMs)
Preparation Method	Macrophages were cultivated in the presence or absence of CL264 for 24?h, before tumor cells were added and co-cultured for 42?h. Radiolabeled thymidine was used to detect tumor cell growth and was added to the co-cultures 18?h before cell harvest.
Reaction Conditions	Macrophages,CL264(40ng/ml),LLC co-cultured for 42?h
Applications	CL264 stimulating the BMDMs and it resulted in inhibition of LLC cells, but only when it was used together with IFN-γ.
Animal experiment [2]:
Animal models	ild-type C57Bl/6J mice,TLR7 Knock out C57Bl/6J mice
Preparation Method	Cancer cells - Murine lung adenocarcinoma LLC-luc cells (1E6 in 100μl PBS/injection) were injected into the flank of recipient mice. Mice then received intra-tumor injections of CL264 at day 0, 3 and 6 after tumor grafting.
Dosage form	40μg/injection, intra-tumor injection
Applications	Intratumoral injection of the TLR7 agonist CL264 induced a pro-tumorigenic effect.CL264 in TLR7 KO mice, similar to that observed in WT mice.These results clearly demonstrate that the pro-tumorigenic effect of CL264 effect is mediated by an effect on TLR7 expressed by malignant (rather than immune) cells.
参考文献: [1]. Müller E, Christopoulos PF, et al. Toll-Like Receptor Ligands and Interferon-γ Synergize for Induction of Antitumor M1 Macrophages. Front Immunol. 2017 Oct 26;8:1383. [2]. Dajon M, Iribarren K, et al. Toll like receptor 7 expressed by malignant cells promotes tumor progression and metastasis through the recruitment of myeloid derived suppressor cells. Oncoimmunology. 2018 Oct 11;8(1):e1505174.