TABLE 4

Effects of a pharmacological modulation of purine system in preclinical models of immune/inflammatory diseases

Experimental Model	Animal	Molecular Target	Ligand	Pharmacological Effect	References
Multiple Sclerosis
Experimental autoimmune encephalomyelitis (EAE)	C57BL/6 mice	P2X₇	oATP (5 or 10 mg/kg/day), Brilliant Blue G (5 or 10 mg/kg/day)	Attenuation of tissue damage and amelioration of neurologic consequences (increase in conduction latency) associated with EAE	Matute et al. (2007)
	C57BL/6 mice	P2Y₁₂	Clopidogrel (5, 15, 50 mg/kg/day), Ticagrelor (30 mg/kg/day)	Amelioration of clinical symptoms	Qin et al. (2017)
				Reduction of leukocyte infiltration in the spinal cord
				Inhibition of Th17 differentiation
	C57BL/6 mice	A_2A	SCH58261 (2 mg/kg/day)	Protection of mice from EAE induction	Mills et al. (2008)
	C57BL/6 mice	A_2A	CGS21680 (0.01 or 0.05 mg/kg/day)	Reduction of the severity of inflammation and tissue damage	Liu et al. (2016)
	C57BL/6 mice	A_2A	CGS21680 (0.01 or 0.05 mg/kg/day)	Reduction of Th1, Th2, and Th17 cells and an increase in T_reg cells, with the reduction of IFN-γ, IL-4, and IL-17 release and the induction of TGF-β release	Liu et al. (2016)
	C57BL/6 mice	A_2A	CGS-21680 (50 µg/kg/day)	Reduction of disease severity in EAE mice	Liu et al. (2018)
				Reduction of spinal cord CD45⁺ cells infiltration
				Decrease of blood-brain barrier permeability
	C57BL/6 mice	A_2B	CVT-6883 (0.3, 1, 3 mg/kg/day), MRS-1754 (1 mg/kg/day)	Reduction of the peak severity and cumulative clinical score	Wei et al. (2013)
	C57BL/6 mice	A_2B	CVT-6883 (0.3, 1, 3 mg/kg/day), MRS-1754 (1 mg/kg/day)	Reduction of the percentage of Th17 and Th1 cells in the CD4⁺ population in the spleen	Wei et al. (2013)
Uveitis
Immunization with the human interphotoreceptor retinoid-binding protein peptide IRBP_1–20	Female C57BL/6 (B6) mice	P2X₇	OxATP i.p. injection of 300 μg/mouse every 3 days	Reduction of Th₁₇ autoreactive T cells	Zhao et al. (2016)
	Female C57BL/6 (B6) mice	Adenosine receptors	NECA i.p. injection of 100 ng/mouse	(0 days postimmunization) suppressive effect on disease development and Th₁₇ responses	Liang et al. (2014)
	Female C57BL/6 (B6) mice	Adenosine receptors	NECA i.p. injection of 100 ng/mouse	(7 days postimmunization) enhanced disease activity and Th₁₇ responses	Liang et al. (2014)
	C57BL/6 J mice	A_2A	CGS 21680 0.5 mg/kg given i.p. once a day for 3 days	Administered at the peak of the disorder accelerated the resolution of disease	Lee et al. (2016)
	C57BL/6 mice	A₃	CF101 10 μg/kg p.o., twice daily for 19 days	Improvement of uveitis clinical scores, amelioration of the pathologic manifestations of the disease and reduction of antigen-specific proliferation and cytokine production of autoreactive T cells	Bar-Yehuda et al. (2011)
	Female C57BL/6 (B6) mice	Endogenous adenosine	ADA 5U/mouse given i.p. for 22 days	Suppression of the course of EAU when given 8–14 days post-immunization and augmentation when given either before or after this period	Liang et al. (2016b)
Myasthenia gravis
Immunization with AChR R97‐116 peptide	Female Lewis rats	A_2A	CGS21680 0.5 mg/kg i.p. every 3 days for 29 days post EAMG induction	Amelioration of disease severity and decrease in the number of Th₁ and Th₂ cells while increasing the number of T_reg cells	Li et al. (2012)
Rheumatoid arthritis
Freund’s adjuvant induced arthritis	DBA/1J mice	P2X₇	Suramin (30 mg/kg), A-438079 (5 mg/kg)	Attenuation of joint damage	Fan et al. (2016)
Freund’s adjuvant induced arthritis	DBA/1J mice	P2X₇	Suramin (30 mg/kg), A-438079 (5 mg/kg)	Reduction of paw edema and IL-17 concentration in synovial fluid	Fan et al. (2016)
Collagen-induced arthritis	C57BL/6 mice	A_2A (pro-drug)	2-(cyclohexylethylthio)adenosine 5′-monophosphate (0.5 mg/kg/min)	Amelioration of clinical and histologic score	Flogel et al. (2012)
				Inhibition of proteo-glycan depletion and cartilage matrix erosion
				Reduction of IL-1, IL-6, IFN-γ, MCP-1, and TNF in synovial fluid
Freund’s adjuvant-induced arthritis	Lewis rats	A₃	1-(methylaminocarbonyl)bicyclo[3.1.0]hexane-2,3-diol] (also named CF 502) (1, 10, and 100 μg/kg)	Amelioration of clinical and histologic score	Ochaion et al. (2008)
				Reduction of PI3K, PKB/AKT, IKK, NF-κB, and TNF in paw extracts
				Reduction of GSK-3β, PARP, and β catenin in paw extracts
Scleroderma
Bleomycin-induced fibrosis	Male C57BL/6 mice	A_2A	ZM241385 (50 mg/kg i.p. twice per day)	Attenuation of bleomycin-induced dermal fibrosis (reduced punch biopsy skin thickness, lower skinfold thickness)	Chan et al. (2006)
	Tcf/Lef:H2B-GFP mice	A_2A	KW6002 (10 mg/kg once per day i.p)	Reduction of skin thickness, skinfold thickness, breaking tension, dermal hydroxyproline content, myofibroblast accumulation, and collagen alignment in bleomycin-induced dermal fibrosis	Zhang et al. (2017a)
	C57BL/6J mice and TSK1 mice	A_2B	C57BL/6J mice: GS-6201 (p.o for 15 days) TSK1 mice: GS-6201(p.o for 30 days)	In C57BL/6J mice: reduction of dermal fibrosis and reduction of extracellular matrix molecule fibronectin and decreased number of alternatively activated macrophages	Karmouty-Quintana et al. (2018)
				In TSK1 mice: reduction of dermal fibrosis at the hyperdermal layer and reduction in hyperdermal layer thickness.
				Reduction of IL-6 and MCP-1 in the skin
Psoriasis
12-Otetradecanoylphorbol-13-acetate	Swiss CD-1	A_2A	CGS-21680 (5 µg per site)	Reduction of epidermal hyperplasia and promotion of collagen synthesis	Arasa et al. (2014)
				Normalization of epidermal structure and enhancement of fibroblast proliferation in the dermis
				Reduction of chemotactic mediator expression and Nfκ-B inhibition
Systemic lupus erythematosus
Genetic model	MRL/lpr mice	A_2A	CGS-21680 (0.4 mg/kg per day, i.p. for 8 wk)	Reduction in proteinuria, blood urea, and creatinine as well as improvement in renal histology	Zhang et al. (2011)
				Reduction of renal macrophage and T-cell infiltration
				Reduction of MCP-1, IFN-γ and MHC-II expression
				Reduction of serum anti-dsDNA and renal immune complex deposition. Inhibition of NFκB activation and suppression the of IFN-γ, MCP-1 and MHC- II expression in splenocytes
Glomerulonephritis
Antibody-mediated glomerulonephritis	Male WKY rats	P2X₇	A-438079 (300 μmol/kg i.p. injection twice daily for 7 days)	Reduction in fibrinoid necrosis	Taylor et al. (2009)
				Reduction in proteinuria
				Reduction in glomerular macrophage infiltration
Genetic model	MRL/lpr mice	P2X₇	brilliant blue G (45.5 mg/kg i.p. injection every 48 h for 8 wk)	Reduction of NLRP3/ASC/caspase 1 assembly, reduction of interleukin-1β release	Zhao et al. (2013)
				Reduction in the severity of nephritis and circulating anti-dsDNA antibodies.
				Reduction of the serum levels of IL-1β and IL-17 and in the Th_l7:T_reg cell ratio
anti-GBM Ab	Male WKY rats	A_2A	CGS 21680 1.5 mg/kg i.p. twice a day for 5 days	Reduction of damage to the kidneys	Garcia et al. (2008)
				Suppression of the glomerular expression of the MDC/CCL22 chemokine and down-regulation of MIP-1α/CCL3, RANTES/CCL5, MIP-1β/CCL4, and MCP-1/CCL2 chemokines
				Increase of anti-inflammatory cytokines, IL-4 and IL-10
Chronic obstructive pulmonary disease
Smoke-induced lung inflammation	C57/Bl6 mice	P2X₇	KN62 (1 μM by mouth 30 min before each cigarette smoke exposure on days 1–3)	Prevention of the lung parenchyma destruction	Lucattelli et al. (2011)
Asthma
Ovalbumin	Balb/c and C57BL/6 mice	P2X₄	5-BDBD (80 μl 100 μM, intratracheally before each of the three consecutive OVA-aerosol challenges)	Reduction of broncho alveolar lavage fluid eosinophilia, peribronchial inflammation, Th2 cytokine production and bronchial hyperresponsiveness	Zech et al. (2016)
	Balb/c and C57BL/6 mice	P2X₇	KN62 (10 μM, intratracheally before allergen challenge)	Reduction of airway eosinophilia, goblet cell hyperplasia, and bronchial hyperresponsiveness to methacholine	Muller et al. (2011)
	Balb/c and C57BL/6 mice	P2X₇	KN62 (10 μM, intratracheally before allergen challenge)	Reduction in allergic airway inflammation	Muller et al. (2011)
	Balb/c and C57BL/6 mice	P2Y₁	MRS2179: 30 mg/kg, MRS2500: 3 mg/kg administered intravenously 20 min before the start of allergen challenge	Reduction of leukocyte recruitment to the lung	Amison et al. (2015)
	Female BALB/c mice	A_2A	CGS-21680 (10 or 100 μg/kg intranasally, half an hour before and 3 h after the challenge)	Inhibition of bronchoalveolar lavage fluid inflammatory cell influx	Bonneau et al. (2006)
	Female BALB/c mice	A_2A		No effect on OVA-induced bronchoconstriction	Bonneau et al. (2006)
Genetical model	ADA-deficient mice	A_2B	CVT-6883 (1 mg/kg i.p. for 14 days)	Reduction of immune cell number in the BAL fluid,	Sun et al. (2006)
				Decreased production of pro-inflammatory cytokines and chemokines
				Attenuation of pulmonary fibrosis
Inflammatory bowel diseases
Trinitrobenzene sulfonic (TNBS) acid	Wistar rats	P2X₇	A740003 (16 mg/kg/day), Brilliant Blue G (40 mg/kg/day)	Amelioration of clinical and histologic scores	Marques et al. (2014)
				Reduction of macrophage and T-cell tissue infiltration
				Reduction of tissue apoptosis
				Inhibition of NF-kappa B and MAP kinase activation
Spontaneous ileitis	SAMP1/YitFc mouse	A_2A	ATL-146e (0.1 μg · kg⁻¹ · min⁻¹)	Decrease of the chronic inflammatory index and villus distortion index	Odashima et al. (2005)
Spontaneous ileitis	SAMP1/YitFc mouse	A_2A	ATL-146e (0.1 μg · kg⁻¹ · min⁻¹)	Reduction of TNF, IFN gamma, and IL- 4 in supernatants from cultures of mesenteric lymph node cells	Odashima et al. (2005)
Oxazolone	Sprague-Dawley rats	A_2A	PSB-0777 (0.4 mg/kg/day)	Amelioration of microscopic damage score	Antonioli et al. (2018a)
Oxazolone	Sprague-Dawley rats	A_2A	PSB-0777 (0.4 mg/kg/day)	Reduction of tissue TNF and oxidative stress	Antonioli et al. (2018a)
Sodium dextran sulfate (DSS)	NMRI mice	A_2A	CGS 21680 (0.5 mg/kg/day)	CGS 21680 was ineffective in ameliorating DSS-induced colitis in mice	Selmeczy et al. (2007)
	C57BL/6 mice	A_2B	ATL-801 (10 mg/kg/day)	Reduction of clinical symptoms, histologic scores, IL-6 levels and proliferation indices	Kolachala et al. (2008a)
	C57BL/6 mice	A_2B	ATL-801 (10 mg/kg/day)	Suppression of the inflammatory infiltrate into colonic mucosa and decrease of epithelial hyperplasia	Kolachala et al. (2008a)
	C57BL/6 mice	A_2B	PSB1115 (1 mg/kg/day)	Increase in severity of DSS colitis	Frick et al. (2009)
	BALB/c mice	A₃	IB MECA (1 or 3 mg/kg/day b.i.d.)	Amelioration of clinical signs of colitis	Mabley et al. (2003)
	BALB/c mice	A₃	IB MECA (1 or 3 mg/kg/day b.i.d.)	Reduction of tissue IL-1, IL-6, IL-12 MIP-1, MIP-2, MPO, and MDA levels	Mabley et al. (2003)
Interleukin-10^−/−	C57BL/6 mice	A₃	IB MECA (1 or 3 mg/kg/day b.i.d.)	Reduction of tissue IL-1, IL-6, MIP-1, MIP-2, MPO, and MDA levels	Mabley et al. (2003)
Trinitrobenzene sulfonic (TNBS) acid	Sprague-Dawley rats	A₃	IB-MECA (1.5 mg/kg b.i.d.)	Improvement of the clinical and histologic score, appetite, and weight gain	Guzman et al. (2006)
Trinitrobenzene sulfonic (TNBS) acid	Sprague-Dawley rats	A₃	IB-MECA (1.5 mg/kg b.i.d.)	Reduction of free radical production	Guzman et al. (2006)
Dinitrobenzene sulfonic (DNBS) acid	Sprague-Dawley rats	Adenosine deaminase	4-amino-2-(2-hydroxy-1-decyl)pyrazole[3,4-d]pyrimidine (APP, 5, 15, or 45 micromol/kg) and erythro-9-(2-hydroxy-3-nonyl)adenine hydrochloride (EHNA, 10, 30, or 90 micromol/kg)	Amelioration of systemic (food intake, body and spleen weight) and colonic [macroscopic/microscopic damage, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and malondialdehyde (MDA)] inflammatory parameters	Antonioli et al. (2007)
Dinitrobenzene sulfonic (DNBS) acid	Sprague-Dawley rats	Adenosine deaminase			Antonioli et al. (2010a)
interleukin-10^−/−	C57BL/6 mice	Adenosine deaminase	Pentostatin (0.75 mg/kg)	Improvement of the clinical and histologic score	Brown et al. (2008a)
interleukin-10^−/−	C57BL/6 mice	Adenosine deaminase	Pentostatin (0.75 mg/kg)	Reduction of Th1 cytokines (IL-1β, IFN-γ, TNF, IL-6, CXCL10)	Brown et al. (2008a)

ATL-801, N-[5-(1-cyclopropyl-2,6-dioxo-3-propyl-2,3,6,7-tetrahydro-1H-purin-8-yl)-pyridin-2-yl]-N-ethyl nicotinamide; ATL 802, N-(5-(1-cyclopropyl-2,6-dioxo-3-propyl-2,3,6,7-tetrahydro-1H-purin-8-yl)pyridin-2-yl)-N-methyl-6-(trifluoromethyl)nicotinamide; CGS 21680, 4-[2-[[6-amino-9-(N-ethyl-β-d-ribofuranuronamidosyl)-9H-purin-yl]amino]ethyl] benzene propanoic acid; MRS2500, (1R,2S,4S,5S)-4-[2-iodo-6-(methylamino)-9H-purin-9-yl]-2 (phosphonooxy)bicyclohexane-1-methanol dihydrogen phosphate ester.