Table 1.

NO₂-FA Evaluation in Preclinical Animal Models^a

Disease state	Animal	Disease model	Dose	Duration	Route of admin	Outcomes	Refs
ALS	Female B6SJL-TgN (SOD1-G93A) 1Gur mice	Onset of disease @ 90 d followed by treatment until 140 d	16 mg/kg/day	7.14 wk	SC injection (3×/wk)	NO2-OA crossed the BBB and was neuroprotective in this ALS model	[80]
Atherosclerosis	8-wk-old apoE−/− mice	Western diet	8 mg/kg/day	3 wk	SC minipump	NO2-OA reduced atherosclerotic lesion formation via decreasing inflammation, macrophage infiltration and limiting the expression of adhesion molecules	[81]
Atrial fibrosis	C57BL/6J mice	AngII infusion via mini pump (1.5 ng/g/min)	6 mg/kg/day	2 wk	SC minipump	NO2-OA decreased the Ang-II-induced fibrotic response in heart	[82]
Atrial fibrosis and fibrillation						NO2-OA decreased Ang-II-induced vulnerability for atrial fibrillation and inhibited atrial fibrosis	[83]
Breast cancer	6-wk-old female athymic nude mice	MDA-MB-231 xenograft tumor growth; NO2-OA started once tumor size was 50–100 mm3	7.5 mg/kg/d	4 wk	Oral gavage	NO2-OA mediates in vivo growth suppression of MDA-MB-231 cells with no overt toxic effects	[72]
Cardiac I/R – MI	8–12-wk-old C57/Bl6 mice	I/R: 30 min unilateral ischemia, 24 h reperfusion	6.6 mg/kg	At time of reperfusion	IP	NO2-OA has protective effects in cardiac I/R resulting in decreased infarct size and improved left ventricular function in all treatment regimens	[30]
				15 min prior to reperfusion	IP
				3 d prior to ischemia	SC minipump
Diabetes	8–10-wk-old C57BL/6J or Lepob (ob/ob) male mice	Genetic model of obesity and insulin resistance on normal chow	8 mg/kg/d	4 wk	SC minipump	NO2-OA normalized hyperglycemia in diabetic mice. Plasma levels of 30 nM were enough to exert a pharmacological effect	[84]
Hypertension	8–10-wk-old C57BL/6J mice	Ang II infusion	5 mg/kg/day	2 wk	SC mini-pump	NO2-OA lowers BP by acting as an antagonist of Ang-II-induced hypertension	[55]
		Pre-existing hypertension: Ang II injection	1.25, 2.5, 5, 10, 20 mg/kg	10 min before or 3 d after Ang II delivery	IV – jugular infusion	Dose-dependent reduction in BP. While PPARγ agonist had no effect on BP reduction by NO2-OA
	C57BL/6	Ang II infusion via mini pump at 1 mg/kg/d	5 mg/kg/d	3 days after Ang II infusion	SC minipump	NO2-OA protects against Ang-II-induced hypertension and is mediated via the inhibition of soluble epoxide hydrolase	[56]
Inflammation (multiorgan sepsis)	8–10-wk-old male C57BL/6 mice	Escherichia coli-induced septic shock (single IP injection of 10 mg/kg); 18 hr	0.2 mg/kg/d	48 h before LPS challenge of 18 h	SC minipump	NO2-OA protects against endotoxin-induced endotoxemia and multiorgan injury in mice	[85]
Inflammation (pulmonary and sepsis)	8-wk-old male C57BL/6/J and 5- LO-deficient mice	LPS (20mg/ kg, IP); 16 h	6.6 mg/kg	1, 4 h before and 4 h after LPS	IP	NO2-OA attenuates LPS-induced neutrophil and monocyte mobilization, irreversibly inhibits 5-LO and inhibits lung injury	[86]
Inflammation (skin)	6–12-wk-old female Balb/c mice	CHS-sensitization with 0.5% DNFB, FITC, or oxazolone	0.84 mg/kg	18 h prior to skin insult	topical	Topical application of NO2-OA augments CHS response	[75]
	and male and female FoxP3DTR mice			18 h prior to skin insult	SC injection	SC injection of NO2-OA inhibits skin inflammation in ACD	[74]
Inflammation (vascular)	C57BL/6J mice	Tail vein injection of LPS (0.5 mg/kg LPS); 3 h	5 mg/kg/d	3 d	SC minipump	NO2-OA decreased vascular inflammation by disrupting TLR4 signaling and inhibiting leukocyte adhesion	[87]
Inflammatory bowel disease	7–8-wk-old female BALB/c mice	DSS-induced (2% in drinking water for 7 d)	0.5 or 5 mg/kg/d	7 d	SC minipump	NO2-OA reduced disease index, prevented colon shortening and the increase in p65 expression by increasing PPARγ expression	[88]
Kidney – diabetic nephropathy	12-wk-old Leprdb/db(db/db) and Leprdb/m (db/m)	Genetic model; coupled NO2-OA with losartan	5 mg/kg/d	2 wk	SC minipump	NO2-OA or losartan alone mildly decreased kidney injury. However, when treatments were combined, the diabetic renal injury was reversed	[89]
Kidney – nephropathy	Male BALB/c mice	ADR-induced nephropathy; mice sacrificed 7 d after 10 mg/kg ADR injection	5 mg/kg/day	2 d before ADR single injection	SC minipump	NO2-OA protects against ADR nephropathy by decreasing inflammation and reactive species generation	[90]
Kidney – Renal I/R	3-mo-old male B6129SF2/J mice	I/R: 30 min warm, bilateral ischemia, 24 h reperfusion	0.5 mg/kg	Starting 1 h after ischemia, every 6 h for 24 h	IP	Delayed administration of NO2-OA attenuates renal I/R injury in the mouse likely via inhibition of the inflammatory response	[91]
NAFLD/NASH	8-wk-old male C57BL/6J and apoE−/− mice	WD (42% fat, TD.88137) NASH (40% fat, D17010103)	5 or 8 mg/kg/d	12 wk	SC minipump	NO2-OA protects against NASH-diet-induced liver damage, hepatomegaly and steatohepatitis	[70]
	6–8-wk-old C57BL/6J mice	HFD (60% fat; D12492)	8 mg/kg/d	6 wk	SC minipump	NO2-OA protects against obesity-induced insulin resistance and steatosis	[92]
Obesity	4-mo-old obese Zucker rats – fa/fa mutation in leptin receptor	Genetic model on normal chow	0.0075 mg/kg/d	2 wk	SC minipump	NO2-OA decreased plasma triglyceride levels, normalizes plasma nonesterified free FAs and increases plasma high-density lipoproteins in obese Zucker rats. NO2-FA may be a safe and effective therapeutic for obesity	[93]
Pulmonary arterial hypertension	8–10-wk-old C57BL/6J mice	Hypoxia (10% O2) for 28 d on normal chow	8 mg/kg/d	2 and 4 wk	SC mini-pump	NO2-OA attenuated hypoxia-induced pulmonary hypertension	[94]
	6–8-wk-old C57BL/6J mice	HFD (60% fat; D12492)	8 mg/kg/d	6.5 wk	SC minipump	NO2-OA protects against obesity-induced insulin resistance and PAH	[95]
Vascular injury	6–8-wk-old C57BL/6 mice	Wire injury of femoral artery	2 mg/kg/d	3 wk	SC minipump	NO2-OA inhibited neointimal hyperplasia after wire-induced femoral artery injury via HO-1-dependent mechanisms	[38]
Ventral hernia	10–12-wk-old female Sprague–Dawley rats	Microparticle delivery of NO2-OA in a ventral hernia rat model	~1200 pmol/scaffold (in vitro assessment)	8 wk	Scaffold implantation	NO2-OA repaired the abdominal wall and increased angiogenesis	[96]

^aAbbreviations: ACD, ; ADR, adriamycin; ALS, amyotrophic lateral sclerosis; BBB, blood–brain barrier; CHS, contact hypersensitivity; DNFB, 1-fluoro-2,4-dinitrobenzene; DSS, dextran sodium sulfate; HFD, high-fat diet; IP, intraperitoneal; IV, intravenous; NAFLD, nonalcoholic fatty liver disease; NASH, nonalcoholic steatohepatitis; LO, lipoxygenase; LPS, lipopolysaccharide; MI, myocardial infarction; SC, subcutaneous; WD, Western diet.