MRE 3008F20 is a highly selective antagonist of the adenosine A3 receptor (AA3R), with a binding constant Ki = 1.8nM[1]. AA3R inhibits adenylyl cyclase and regulates intracellular calcium levels, playing a key role in cardioprotection, anti-ischemia, and anti-inflammation[2]. MRE 3008F20 is commonly used in radioligand binding assays, in vitro pharmacology (e.g., receptor binding experiments and functional inhibition assays), and studies of related signaling pathways[3,4].
In vitro, pretreatment of mucosal-type bone-marrow-derived mast cells (mBMMCs) with MRE 3008F20 (10μM) for 20min, followed by stimulation with adenosine (0.1 or 1μM), significantly inhibited the adenosine-induced increase in intracellular calcium. Under stimulation with 0.1μM adenosine, the activation rate decreased from 2.7 ± 0.3% to 1.3 ± 0.2% (p < 0.05), and under stimulation with 1μM adenosine, the activation rate decreased from 9.0 ± 0.7% to 1.8 ± 0.3% (p < 0.01)[5]. Treatment of human melanoma A375 cells with MRE 3008F20 (1μM) in combination with 10μM VP-16 or 1μM doxorubicin for 4h further enhanced the inhibitory effects of VP-16 and doxorubicin on HIF-1α protein expression[6]. Pretreatment of human melanoma C32 cells with MRE 3008F20 (10nM) for 30min, followed by treatment with adenosine deaminase (ADA, 0.3U/mL) and continued culture for 48h, completely blocked the cell proliferation induced by ADA[7].
References:
[1] GESSI S, VARANI K, MERIGHI S, et al. Expression of A3 adenosine receptors in human lymphocytes: up-regulation in T cell activation[J]. Molecular Pharmacology, 2004, 65(3): 711-719.
[2] DAL BEN D, LAMBERTUCCI C, MARUCCI G, et al. Adenosine receptor modeling: what does the A2A crystal structure tell us?[J]. Current Topics in Medicinal Chemistry, 2010, 10(10): 993-1018.
[3] VARANI K, MERIGHI S, GESSI S, et al. [3H] MRE 3008F20: a novel antagonist radioligand for the pharmacological and biochemical characterization of human A3 adenosine receptors[J]. Molecular Pharmacology, 2000, 57(5): 968-975.
[4] GESSI S, VARANI K, MERIGHI S, et al. A3 adenosine receptors in human neutrophils and promyelocytic HL60 cells: a pharmacological and biochemical study[J]. Molecular Pharmacology, 2002, 61(2): 415-424.
[5] YASHIRO T, OGATA H, ZAIDI S F, et al. Pathophysiological roles of neuro-immune interactions between enteric neurons and mucosal mast cells in the gut of food allergy mice[J]. Cells, 2021, 10(7): 1586.
[6] MERIGHI S, SIMIONI C, GESSI S, et al. A2B and A3 adenosine receptors modulate vascular endothelial growth factor and interleukin-8 expression in human melanoma cells treated with etoposide and doxorubicin[J]. Neoplasia, 2009, 11(10): 1064-1073.
[7] SOARES A S, COSTA V M, DINIZ C, et al. Inosine Strongly Enhances Proliferation of Human C32 Melanoma Cells through PLC‐PKC‐MEK 1/2‐ERK 1/2 and PI3K Pathways[J]. Basic & Clinical Pharmacology & Toxicology, 2015, 116(1): 25-36.
MRE 3008F20是一种具有高效选择性的腺苷A3受体（AA3R）拮抗剂，结合常数Ki = 1.8nM[1]。AA3R能抑制腺苷酸环化酶并调节细胞内钙水平，在心脏保护、抗缺血和抗炎等方面发挥关键作用[2]。MRE 3008F20通常用于放射性配体结合测定、体外药理学（如受体结合实验和功能抑制实验）和相关信号传导的研究[3,4]。
在体外，MRE 3008F20（10μM）预处理黏膜型骨髓源性肥大细胞mBMMCs 20min，加入adenosine（0.1 或 1μM）刺激后，显著抑制了由 adenosine 诱导的细胞内钙离子升高，在 0.1μM adenosine 刺激下激活率从 2.7 ± 0.3% 降至 1.3 ± 0.2%（p < 0.05），在 1μM adenosine 刺激下激活率从 9.0 ± 0.7% 降至 1.8 ± 0.3%（p < 0.01）[5]。MRE 3008F20（1μM）与10μM VP-16或1μM doxorubicin联合处理人黑色素瘤A375细胞4h，可进一步增强VP-16和doxorubicin对HIF-1α蛋白表达的抑制作用[6]。MRE 3008F20（10nM）预处理人黑色素瘤细胞C32 30min，加入adenosine deaminase（ADA, 0.3U/mL）处理后继续培养48h，完全阻断了由ADA诱导的细胞增殖[7]。