SSR-240612 HCl 464930-42-5_Bradykinin Receptor_GPCR & G Protein_Signaling Pathways_Products

SSR-240612 HCl, the hydrochloride salt of SSR240612, is a novel, potent, and orally bioactive specific non-peptide bradykinin B(1) receptor antagonist. SSR-240612 has the potential for the treatment of inflammation and chronic pain. In human fibroblast MRC5 and recombinant human B(1) receptor expressed in human embryonic kidney cells, SSR240612 inhibits the binding of [(3)H]Lys(0)-des-Arg(9)-BK to the B(1) receptor. The inhibition constants (K(i)) of these two receptors are 0.48 and 0.73 nM, respectively. With an IC(50) of 1.9 nM, SSR240612 prevented human fibroblast MRC5 from forming inositol monophosphate in response to Lys(0)-desAr(9)-BK (10 nM). Also, it inhibited the des-Arg(9)-BK-induced contractions in the rat ileum's mesenteric plexus and isolated rabbit aorta, with pA(2) values of 8.9 and 9.4, respectively.

Physicochemical Properties

Molecular Formula

C42H53CLN4O7S

Molecular Weight

793.42

Exact Mass

792.332

Elemental Analysis

C, 63.58; H, 6.73; Cl, 4.47; N, 7.06; O, 14.12; S, 4.04

CAS #

464930-42-5

Related CAS #

465539-70-2

PubChem CID

44235958

Appearance

White to off-white solid powder

LogP

9.395

Hydrogen Bond Donor Count

Hydrogen Bond Acceptor Count

Rotatable Bond Count

Heavy Atom Count

Complexity

1330

Defined Atom Stereocenter Count

SMILES

Cl.S(C1C=CC2C=C(C=CC=2C=1)OC)(N[C@@H](C1=CC=C2C(=C1)OCO2)CC(N[C@@H](C(N(C)C(C)C)=O)CC1C=CC(=CC=1)CN1[C@H](C)CCC[C@@H]1C)=O)(=O)=O

InChi Key

GLHHFOSVBQQNAW-GDYXXZBVSA-N

InChi Code

InChI=1S/C42H52N4O7S.ClH/c1-27(2)45(5)42(48)38(20-30-10-12-31(13-11-30)25-46-28(3)8-7-9-29(46)4)43-41(47)24-37(34-16-19-39-40(23-34)53-26-52-39)44-54(49,50)36-18-15-32-21-35(51-6)17-14-33(32)22-36;/h10-19,21-23,27-29,37-38,44H,7-9,20,24-26H2,1-6H3,(H,43,47);1H/t28-,29+,37-,38-;/m1./s1

Chemical Name

(2R)-2-[[(3R)-3-(1,3-benzodioxol-5-yl)-3-[(6-methoxynaphthalen-2-yl)sulfonylamino]propanoyl]amino]-3-[4-[[(2R,6S)-2,6-dimethylpiperidin-1-yl]methyl]phenyl]-N-methyl-N-propan-2-ylpropanamide;hydrochloride

Synonyms

SSR240612; SSR-240612 hydrochloride; SSR 240612; SSR240612 HCl

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

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

bradykinin B1 receptor, Human MRC5 ( Ki = 0.48 nM );
bradykinin B1 receptor, Human HEK-B1 ( IC50 = 0.73 nM );
bradykinin B2 receptor, guinea pig ileum membranes ( IC50 = 481 nM );
bradykinin B2 receptor, Human CHO-B2 ( IC50 = 358 nM )

ln Vitro

In vitro activity: SSR240612 is a strong antagonist of the bradykinin B1 receptor, with Kis values of 0.48 nM and 0.73 nM for the B2 kinin receptors of human fibroblast MRC5 and HEK cells that express human B1 receptors, respectively, and 481 nM and 358 nM for the B1 receptors of guinea pig ileum membranes and CHO cells that express human B1 receptor. The human fibroblast MRC5 is activated by BK (3 nM) to produce inositol phosphate-1, but SSR240612 inhibits this formation with an IC50 of 1.9 nM and has no discernible effect on this process[1].

ln Vivo

SSR240612 (10 mg/kg p.o. or 0.3, 1 mg/kg i.p.) clearly prevents the mice's paw edema caused by des-Arg9-BK. In the formalin model of inflammation in mice, SSR240612 (10 and 30 mg/kg) reduces, in a dose-dependent manner, the duration of the late phase of paw licking. The administration of SSR240612 (0.3, 3, and 30 mg/kg, p.o.) prior to capsaicin therapy potently and independently of concentration decreases ear edema. Following splanchnic artery occlusion/reperfusion, SSR240612 (0.3 mg/kg, i.v.) also inhibits tissue deterioration and neutrophil accumulation in the rat colon. Furthermore, SSR240612 (1 and 3 mg/kg p.o.) dramatically lengthens the withdrawal latencies in rats experiencing UV-induced thermal hyperalgesia[1]. In rats fed glucose, SSR240612 attenuates tactile and cold allodynia at 3 hours, but it had no effect on control rats, whose ID50s were 5.5 and 7.1 mg/kg, respectively. In rats given glucose at a dose of 10 mg/kg, SSR240612 has no effect on insulin resistance (HOMA index), plasma glucose and insulin, or the generation of aortic superoxide anion[2].

Enzyme Assay

[³H]Lys0-des-Arg9-BK The following ingredients make up the binding buffer used for binding to cell membranes: 137 mM NaCl, 5.4 mM KCl, 1.05 mM MgCl₂, 1.8 mM CaCl₂, 1.2 mM NaH₂PO₄, 15.5 mM NaHCO₃, 10 mM HEPES, 1 g/L bovine serum albumin (BSA), 140 mg/L bacitracin, and 1 μM captopril, pH 7.4. The membranes are incubated in 500 μL of binding buffer containing 1 nM [³H]Lys0-des-Arg9-BK for 30 minutes at 25°C. for saturation isotherms and 0.1 to 10 nM for competition curves. Filters are washed three times with 5 mL of binding buffer, and radioactivity is determined by liquid scintillation spectrometry. One way to measure nonspecific binding is to add 1 μM of unlabeled Lys0-des-Arg9BK[1].

Cell Assay

SSR240612 is a strong antagonist of the bradykinin B1 receptor, with Kis values of 0.48 nM and 0.73 nM for the B2 kinin receptors of human fibroblast MRC5 and HEK cells that express human B1 receptors, respectively, and 481 nM and 358 nM for the B1 receptors of guinea pig ileum membranes and CHO cells that express human B1 receptor. SSR240612 inhibits the formation of inositol phosphate-1 with an IC50 of 1.9 nM, but has no discernible effect on the formation of inositol phosphate-1 in human fibroblast MRC5 that is triggered by BK (3 nM) activation of the B2 receptor.

Animal Protocol

Mice: Groups of eight male albino mice are given an intraplantar injection (20 μL) containing 5 μg of IL-1β in phosphate-buffered saline/0.1% BSA into their right hind paw while they are under isoflurane anesthesia. Subject to anesthesia, mice are given a 20-μL intraplantar injection of des-Arg9-BK (10 μg/paw) in water forty minutes later (T = 0). Acute administration of SSR240612 or vehicle (5% (v/v) ethanol and 5% (v/v) Tween 80 in water) is done by intraperitoneal route 40 minutes prior to des-Arg9-BK injection, and orally at doses of 1, 3, and 10 mg/kg one hour before the injection. Paw volume is measured using a plethysmometer at T = -2 hours (first measurement) and at multiple intervals (T = 20, 40, 60, and 120 min) following edema induction. The difference between the initial paw volume and the paw volume at each time after edema induction is the paw edema volume, measured in milliliters. The mean ± S.E.M. of the individual paw edema volumes is presented for each group. After confirming the normality and homogeneity of variances with multiple ANOVA tests, Duncan's test is used to compare the treated groups to the des-Arg9-BK control group[1]. Rats: SSR240612 (suspended with 0.1% Tween 80 in saline) is given orally, two hours prior to the thermal hyperalgesia measurement, at a dose of 20 milliliters per kilogram. Prior to measuring the rats' withdrawal latencies in the time course study, the compound is given orally at a dose of 3 mg/kg at intervals of 0.08, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. The results are given as mean withdrawal latencies (s) ± S.E.M. in seconds, and statistical analyses are carried out with a two-way ANOVA and Dunnett's test[1].

References

[1]. SSR240612 [(2R)-2-[((3R)-3-(1,3-benzodioxol-5-yl)-3-[[(6-methoxy-2-naphthyl)sulfonyl]amino]propanoyl)amino]-3-(4-[[2R,6S)-2,6-dimethylpiperidinyl]methyl]phenyl)-N-isopropyl-N-methylpropanamide hydrochloride], a new nonpeptide antagonist of the bradykinin B1 receptor: biochemical and pharmacological characterization. J Pharmacol Exp Ther . 2004 May;309(2):661-9

[2]. The kinin B1 receptor antagonist SSR240612 reverses tactile and cold allodynia in an experimental rat model of insulin resistance. Br J Pharmacol . 2007 Sep;152(2):280-7.

Solubility Data

Solubility (In Vitro)

DMSO: ≥ 100 mg/mL (~126.0 mM)

Water: N/A

Ethanol: N/A

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (3.15 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.

Solubility in Formulation 2: ≥ 2.5 mg/mL (3.15 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 25.0 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: ≥ 2.5 mg/mL (3.15 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.

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

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.2604 mL	6.3018 mL	12.6037 mL
	5 mM	0.2521 mL	1.2604 mL	2.5207 mL
	10 mM	0.1260 mL	0.6302 mL	1.2604 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

SSR-240612 HCl 464930-42-5

CAS No.: 464930-42-5

Physicochemical Properties

Biological Activity

Solubility Data