Advanced glycation end product (AGE) formation (IC₅₀ = 275 µM for inhibition of metal ion-catalyzed ascorbate oxidation) [1]

LR-90 dose-dependently reduced the expression of the genes for RAGE, MCP-1, COX-2, IP-10, and NOX2 after 1 hour of pretreatment. Four hours prior to S100b stimulation [1]. After 24 hours of pretreatment, 1 hour before to S100b stimulation, LR-90 (0, 25, 50, 100, and 200 μM) substantially and dose-dependently inhibited the adherence of THP-1 cells to endothelial cells [1]. THP-1 cell viability was unaffected by LR-90 (0, 25, 50, 100, and 200 μM) for a 24-hour period [1].

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- LR-90 pretreatment (1 hour) dose-dependently reduced mRNA expression of RAGE, MCP-1, COX-2, IP-10, and NOX2 in human monocytes stimulated with S100B [1]
- Pretreatment with LR-90 (0, 25, 50, 100, 200 µM) for 24 hours, followed by 1 hour of S100B stimulation, dose-dependently inhibited THP-1 cell adhesion to endothelial cells [1]
- LR-90 (0–200 µM) did not affect THP-1 cell viability after 24-hour exposure [1]

LR-90 (50 mg/L, orally administered for 27 weeks) considerably decreases plasma lipids and has a certain influence on hyperglycemia in ZDF rats [2]. LR-90 (50 mg/L) decreases renal AGE, AGER and lipid peroxidation [2].

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- Oral administration of LR-90 (50 mg/L in drinking water) for 27 weeks significantly reduced plasma triacylglycerol by ~55% and cholesterol by ~30% in Zucker diabetic fatty (ZDF) rats [2]
- LR-90 (50 mg/L) decreased renal AGE accumulation, AGE receptor (AGER) expression, and lipid peroxidation in ZDF rats [2]
- Mild improvements in hyperglycemia and blood pressure, and reduced body weight were observed in LR-90-treated ZDF rats [2]

Assay for metal ion-catalyzed ascorbate oxidation inhibition: LR-90 was tested for its ability to scavenge dicarbonyl intermediates and chelate transition metals involved in AGE formation, with an IC₅₀ of 275 µM [1]

Cell viability assay [1]
Cell Types: THP-1 Cell
Tested Concentrations: 0, 25, 50, 100 and 200 μM
Incubation Duration: 24 hrs (hours)
Experimental Results: No cytotoxicity to THP-1 cells.

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RT-PCR[1]
Cell Types: THP-1 Cell
Tested Concentrations: 0, 25, 50, 100 and 200 μM
Incubation Duration: 1 hour before adding S100b, for 4 hrs (hours)
Experimental Results: Dose-dependent inhibition of RAGE, MCP-mRNA expression1 , COX-2, IP-10 and NOX2 were stimulated with S100b.

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- THP-1 cell adhesion assay: Cells were pretreated with LR-90 (0, 25, 50, 100, 200 µM) for 24 hours, then stimulated with S100B for 1 hour. Adhesion to endothelial cells was quantified [1]
- mRNA expression analysis: Human monocytes were pretreated with LR-90 for 1 hour, stimulated with S100B, and gene expression of RAGE, MCP-1, COX-2, IP-10, and NOX2 was measured by PCR [1]

Animal/Disease Models: Male ZDF rats (13 to 40 weeks) [2]
Doses: 50 mg/L
Route of Administration: Oral administration for 27 weeks.
Experimental Results: Plasma triacylglycerol and cholesterol were Dramatically diminished by approximately 55% and approximately 30% respectively. Mild effects on hyperglycemia and blood pressure. Lowered body weight.

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ZDF rats were administered LR-90 at 50 mg/L via drinking water for 27 weeks. Body weight, blood glucose, blood pressure, plasma lipids, and renal parameters were monitored periodically [2]

[1]. Anti-inflammatory effects of the advanced glycation end product inhibitor LR-90 in human monocytes. Diabetes. 2007 Mar;56(3):647-55.

[2]. LR-90 prevents dyslipidaemia and diabetic nephropathy in the Zucker diabetic fatty rat. Diabetologia. 2008 May;51(5):882-91.

LR-90 is an AGE inhibitor that inhibits S100B and TNFα-induced NF-κB activation [1]. LR-90 has a protective effect against diabetic nephropathy, which is related to the accumulation of collagen and reduced cross-linking in renal tissue [2]. The binding of advanced glycation end products (AGE) to their receptor (RAGE) plays an important role in the development of various diabetic complications, including atherosclerosis. Activation, adhesion, and migration of monocytes are key events in the pathogenesis of atherosclerosis. Previous studies have shown that S100b, a specific ligand of AGE and RAGE, can enhance the inflammatory response of monocytes through RAGE. This study investigated whether a novel AGE inhibitor, LR-90, could inhibit the inflammatory response of human monocytes. We pretreated human THP-1 cells with LR-90 and then stimulated the cells with S100b. LR-90 significantly inhibited S100b-induced RAGE and other pro-inflammatory gene expression, including monocyte chemoattractant protein-1, interferon-γ-induced protein-10, and cyclooxygenase-2, in a dose-dependent manner. These inhibitory effects likely derive from suppressing nuclear factor-κB (NF-κB) activation, as LR-90 inhibits S100b and tumor necrosis factor-α-induced IκB-α degradation and NF-κB promoter transcriptional activity. LR-90 also suppressed oxidative stress in activated monocytes, as evidenced by its inhibition of S100b-induced NADPH oxidase expression and intracellular superoxide production. Furthermore, LR-90 blocked S100b-induced adhesion of monocytes to human umbilical vein endothelial cells. These new data suggest that LR-90 possesses novel anti-inflammatory properties in addition to its AGE-inhibiting effects, and therefore may offer additional protection against diabetic vascular complications. [1]

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Objective/Hypothesis: Previous studies have shown that the novel AGE generation inhibitor LR-90 can prevent the development of experimental type 1 diabetic nephropathy. This study aimed to investigate the role of LR-90 in Zucker diabetic fat (ZDF) rats (a model of type 2 diabetes and metabolic syndrome) and to study its mechanism of protecting the kidneys from damage. Methods: Male ZDF rats were treated with or not treated with LR-90 from 13 to 40 weeks of age. Metabolic function, renal function, and renal histology were assessed. AGE accumulation and AGE receptor (AGER) generation were detected. Fibrotic growth factors, extracellular matrix proteins, and intracellular signaling pathways associated with glomerular and tubular damage were also analyzed. Results: LR-90 significantly reduced plasma lipid levels in ZDF rats, with little effect on hyperglycemia. LR-90 reduced the levels of advanced glycation end products (AGE), advanced glycation end products (AGER), and lipid peroxidation in the kidneys. LR-90 significantly delayed the increase in albuminuria and proteinuria. This was associated with a reduction in glomerular sclerosis and tubulointerstitial fibrosis, while the excessive production of transforming growth factor-β1 (TGF-β1), connective tissue growth factor, fibronectin and type IV collagen in the kidneys was also significantly inhibited. In addition, LR-90 downregulated the activation of key mitogen-activated protein kinase (MAPK) and nuclear factor κB (NF-κB) in the renal cortex. Conclusion/Interpretation: These results support our previous research on the renal protective effect of LR-90 on type 1 diabetic nephropathy and further demonstrate that LR-90 (a pleiotropic AGE inhibitor) may also be beneficial in the prevention of type 2 diabetic nephropathy, in which multiple risk factors such as hyperglycemia, dyslipidemia, obesity, insulin resistance and hypertension can lead to kidney damage. [2]

C35H34CL2N4O8

709.5725

708.175

245075-84-7

10212249

Typically exists as White to off-white solid at room temperature

8.529

6

8

12

49

1060

0

ClC1=C(C([H])=C([H])C(=C1[H])C([H])([H])C1C([H])=C([H])C(=C(C=1[H])Cl)N([H])C(N([H])C1C([H])=C([H])C(=C([H])C=1[H])OC(C(=O)O[H])(C([H])([H])[H])C([H])([H])[H])=O)N([H])C(N([H])C1C([H])=C([H])C(=C([H])C=1[H])OC(C(=O)O[H])(C([H])([H])[H])C([H])([H])[H])=O

WJDMCXSGUCNRPZ-UHFFFAOYSA-N

InChI=1S/C35H34Cl2N4O8/c1-34(2,30(42)43)48-24-11-7-22(8-12-24)38-32(46)40-28-15-5-20(18-26(28)36)17-21-6-16-29(27(37)19-21)41-33(47)39-23-9-13-25(14-10-23)49-35(3,4)31(44)45/h5-16,18-19H,17H2,1-4H3,(H,42,43)(H,44,45)(H2,38,40,46)(H2,39,41,47)

2-[4-[[4-[[4-[[4-(2-carboxypropan-2-yloxy)phenyl]carbamoylamino]-3-chlorophenyl]methyl]-2-chlorophenyl]carbamoylamino]phenoxy]-2-methylpropanoic acid

245075-84-7; LR-90; PJ8V885848; RefChem:154162; 2,2'-(4,4'-(4,4'-methylenebis(2-chloro-4,1-phenylene))bis(azanediyl)bis(oxomethylene)bis(azanediyl)bis(4,1-phenylene))bis(oxy)bis(2-Methylpropanoic acid); lR90; 2,2'-((((((methylenebis(2-chloro-4,1-phenylene))bis(azanediyl))bis(carbonyl))bis(azanediyl))bis(4,1-phenylene))bis(oxy))bis(2-methylpropanoic acid); 2,2'-(Methylenebis((2-chloro-4,1-phenylene)iminocarbonylimino-4,1-phenyleneoxy))bis(2-methylpropanoic acid);

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ≥ 100 mg/mL (~140.93 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (3.52 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (3.52 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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 (Please use freshly prepared in vivo formulations for optimal results.)