Bindarit (AF2838) 130641-38-2_CCR_GPCR & G Protein_Signaling Pathways_Products

Bindarit (also known as AF-2838; AF 2838) is a potent and selective inhibitor of monocyte chemotactic proteins MCP-1/CCL2, MCP-3/CCL7 and MCP-2/CCL8 with anti-inflammatory activity. Bindarit treatment inhibits human monocytes' ability to produce monocyte chemotactic protein-1 (MCP-1) in response to 403 µM and 172 µM, respectively, when exposed to bacterial LPS or Candida albicans. This effect is dose-dependent. With an IC50 of 75 µM, lower levels of MCP-1 mRNA transcripts are linked to Bindarit's inhibition of LP-induced MCP-1 production. When LPS-stimulated MM6 cells are exposed to bindarit, their production of MCP-1 is inhibited (IC50 = 425 μM), but neither IL-8 nor IL-6 are released.

Physicochemical Properties

Molecular Formula

C19H20N2O3

Molecular Weight

324.37

Exact Mass

324.147

Elemental Analysis

C, 70.35; H, 6.21; N, 8.64; O, 14.80

CAS #

130641-38-2

Related CAS #

130641-38-2

PubChem CID

71354

Appearance

White to off-white solid powder

Density

1.2±0.1 g/cm3

Boiling Point

542.9±40.0 °C at 760 mmHg

Flash Point

282.1±27.3 °C

Vapour Pressure

0.0±1.5 mmHg at 25°C

Index of Refraction

1.595

LogP

3.44

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

434

Defined Atom Stereocenter Count

SMILES

O(CC1C2C=CC=CC=2N(CC2C=CC=CC=2)N=1)C(C(=O)O)(C)C

InChi Key

MTHORRSSURHQPZ-UHFFFAOYSA-N

InChi Code

InChI=1S/C19H20N2O3/c1-19(2,18(22)23)24-13-16-15-10-6-7-11-17(15)21(20-16)12-14-8-4-3-5-9-14/h3-11H,12-13H2,1-2H3,(H,22,23)

Chemical Name

2-[(1-benzylindazol-3-yl)methoxy]-2-methylpropanoic acid

Synonyms

AF-2838; AF2838; Bindarit; AF 2838

HS Tariff Code

2934.99.9001

Storage

Powder-20°C 3 years

4°C 2 years

In solvent -80°C 6 months

-20°C 1 month

Shipping Condition

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

Biological Activity

Targets

MCP-1/CCL2; MCP-3/CCL7; MCP-2/CCL8

ln Vitro

In vitro activity: Bindarit treatment inhibits human monocytes' ability to produce monocyte chemotactic protein-1 (MCP-1) in response to 403 µM and 172 µM, respectively, when exposed to bacterial LPS or Candida albicans. This effect is dose-dependent. With an IC50 of 75 µM, lower levels of MCP-1 mRNA transcripts are linked to Bindarit's inhibition of LP-induced MCP-1 production. When LPS-stimulated MM6 cells are exposed to bindarit, their production of MCP-1 is inhibited (IC50 = 425 μM), but neither IL-8 nor IL-6 are released.[2]
Bindarit treatment prevents the IL-1-stimulated osteoblast cell line Saos-2 from releasing MCP-1.[3]
Bindarit, does not directly cause cytotoxicity in vitro in human IIB-MEL-J melanoma or ECs, even at the highest concentration; however, it does suppress MCP-1 expression.[4] Bindarit (10-300 μM) inhibits the proliferation, migration, and invasion of rat vascular smooth muscle cells (VSMCs).[5]
Bindarit causes the traditional NF-κB pathway to be downregulated. With no effect on other tested activated promoters, Bindarit specifically inhibits the activation of the p65 and p65/p50 induced MCP-1 promoter. This suggests that Bindarit selectively targets a subpopulation of NF-κB isoforms within the entire NF-κB inflammatory pathway.[6]
Bindarit primarily regulates TGF-β and AKT signaling negatively in order to modify the migration and proliferation of cancer cells.[7]

ln Vivo

In mice with lupus or NZB/W backgrounds, oral administration of Bindarit at a dose of 50 mg/kg significantly protects against impairment of renal function and delays the onset of proteinuria. MCP-1 up-regulation during the course of nephritis is fully inhibited by bindarit treatment.[1] In human melanoma xenografts, inhibition of MCP-1 with Bindarit yields necrotic tumor masses by reducing tumor growth and macrophage recruitment.[4] Bindarit has a direct impact on VSMC proliferation and migration, as well as neointimal macrophage content, which effectively reduces the formation of neointima in both hyperlipidemic and non-hyperlipidemic animal models of vascular injury.[5] When Balb/c mice with murine breast cancer 4T1-Luc cells are administered Bindarit, the mice's ability to develop local tumors is reduced and their prostate cancer PC-3M-Luc2 xenograft mice showed less metastatic disease. Furthermore, the administration of Bindarit considerably reduces the infiltration of myeloid-derived suppressor cells and tumor-associated macrophages in 4T1-Luc primary tumors.[7]

Cell Assay

In a dose-dependent manner, bindarit inhibited the production of MCP-1 and TNF-alpha in monocytes induced by LPS and Candida albicans, with IC50 values of 172 µM and 403 µM, respectively.With an IC50 value of 75 µM, lower levels of MCP-1 mRNA transcripts have been linked to Bindarit's inhibition of LP-induced MCP-1 production. Without influencing the release of IL-8 or IL-6, bindaritex demonstrated an IC50 of 425 μM inhibitory effect on the production of MCP-1 by LPS-stimulated MM6 cells[3]. Rat vascular smooth muscle cell (VSMC) invasion, migration, and proliferation were all decreased by bindarit (10–300 μM) administration.

Animal Protocol

Formulated in 0.5% carboxy-methylcellulose; ~50 mg/kg/day; Oral gavage

NZB/W F1 female mice with a spontaneous autoimmune disease

References

[1]. Kidney Int . 1998 Mar;53(3):726-34.

[2]. Eur Cytokine Netw . 1999 Sep;10(3):437-42.

[3]. Inflamm Res . 2002 May;51(5):252-8.

[4]. J Invest Dermatol . 2007 Aug;127(8):2031-41.

[5]. Cardiovasc Res . 2009 Dec 1;84(3):485-93.

[6]. Cell Cycle . 2012 Jan 1;11(1):159-69.

[7]. Clin Exp Metastasis . 2012 Aug;29(6):585-601.

Additional Infomation

Bindarit has been used in trials studying the prevention and treatment of Coronary Restenosis and Diabetic Nephropathy.

Solubility Data

Solubility (In Vitro)

DMSO: ~65 mg/mL (~200.4 mM)

Water: <1 mg/mL

Ethanol: ~65 mg/mL (~200.4 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.67 mg/mL (8.23 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 26.7 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.

Solubility in Formulation 2: ≥ 2.67 mg/mL (8.23 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 26.7 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

Solubility in Formulation 3: 2.67 mg/mL (8.23 mM) 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 26.7 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

Solubility in Formulation 4: 0.5% CMC: 7 mg/mL

Solubility in Formulation 5: 5 mg/mL (15.41 mM) in 50% PEG300 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	3.0829 mL	15.4145 mL	30.8290 mL
	5 mM	0.6166 mL	3.0829 mL	6.1658 mL
	10 mM	0.3083 mL	1.5414 mL	3.0829 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

PeptideDB

Bindarit (AF2838) 130641-38-2

CAS No.: 130641-38-2

Physicochemical Properties

Biological Activity

Solubility Data