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 )
SSR-240612 HCl targets bradykinin B1 receptor (BK B1 receptor); Ki = 0.48 nM (human fibroblast MRC5 B1 receptor), Ki = 0.73 nM (recombinant human B1 receptor expressed in HEK cells); selectivity for B1 vs. B2 receptors is 500- to 1000-fold [1]
SSR-240612 HCl targets bradykinin B1 receptor (BK B1 receptor) [2]

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].

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1. Receptor binding inhibition: SSR-240612 HCl potently inhibited the binding of [(3)H]Lys(0)-des-Arg(9)-BK to bradykinin B1 receptors in human fibroblast MRC5 cells (Ki=0.48 nM) and recombinant human B1 receptors expressed in human embryonic kidney (HEK) cells (Ki=0.73 nM) [1]
2. Functional inhibition of B1 receptor signaling: SSR-240612 HCl inhibited Lys(0)-desArg(9)-BK (10 nM)-induced inositol monophosphate formation in human fibroblast MRC5 cells, with an IC50 of 1.9 nM [1]
3. Ex vivo tissue contraction inhibition: SSR-240612 HCl antagonized des-Arg(9)-BK-induced contractions of isolated rabbit aorta (pA2=8.9) and mesenteric plexus of rat ileum (pA2=9.4) [1]

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].

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1. Inhibition of inflammatory edema in mice: SSR-240612 HCl inhibited des-Arg(9)-BK-induced paw edema in mice at doses of 3 and 10 mg/kg (oral administration) and 0.3 and 1 mg/kg (intraperitoneal administration); it also reduced capsaicin-induced ear edema in mice at oral doses of 0.3, 3 and 30 mg/kg [1]
2. Attenuation of intestinal injury in rats: SSR-240612 HCl (0.3 mg/kg, intravenous administration) reduced tissue destruction and neutrophil accumulation in the rat intestine following splanchnic artery occlusion/reperfusion [1]
3. Analgesic effects in rats: SSR-240612 HCl inhibited thermal hyperalgesia induced by UV irradiation (1 and 3 mg/kg, oral) and the late phase of nociceptive response to formalin (10 and 30 mg/kg, oral) in rats; it also prevented neuropathic thermal pain induced by sciatic nerve constriction in rats at oral doses of 20 and 30 mg/kg [1]
4. Reversal of allodynia in insulin-resistant rats: In rats fed with 10% D-glucose for 12 weeks (insulin resistance model with tactile and cold allodynia), orally administered SSR-240612 HCl (0.3-30 mg/kg) blocked tactile allodynia (ID50=5.5 mg/kg at 3 h post-administration) and cold allodynia (ID50=7.1 mg/kg at 3 h post-administration); the compound had no effect on tactile/cold allodynia in age-matched control rats, and 10 mg/kg oral dose had no impact on plasma glucose/insulin levels, HOMA index (insulin resistance marker), or aortic superoxide anion production in glucose-fed rats [2]

[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].

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1. Bradykinin B1 receptor binding assay in human fibroblast MRC5 cells: Membrane preparations from MRC5 cells were incubated with [(3)H]Lys(0)-des-Arg(9)-BK (specific radioligand for B1 receptor) and serial concentrations of SSR-240612 HCl (range not specified) at 25°C for a predetermined incubation time (not specified); non-specific binding was determined in the presence of excess unlabeled Lys(0)-des-Arg(9)-BK; bound radioactivity was separated by filtration and quantified using liquid scintillation counting; Ki values were calculated using standard ligand binding equations [1]
2. Bradykinin B1 receptor binding assay in recombinant HEK cells: Membranes from HEK cells expressing recombinant human B1 receptors were prepared and incubated with [(3)H]Lys(0)-des-Arg(9)-BK and serial dilutions of SSR-240612 HCl under the same conditions as the MRC5 cell assay; bound radioactivity was measured, and Ki values were computed to assess the affinity of SSR-240612 HCl for recombinant B1 receptors [1]
3. Inositol monophosphate formation assay in MRC5 cells: MRC5 cells were seeded in 24-well plates and preincubated with SSR-240612 HCl (serial concentrations) for 15 minutes at 37°C; Lys(0)-desArg(9)-BK (10 nM) was then added to stimulate B1 receptor signaling, and cells were incubated for an additional period (not specified); intracellular inositol monophosphate levels were extracted and quantified using a radiometric assay (method not specified); IC50 values were determined from concentration-response curves [1]

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.

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].\nRats: 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].

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\n1. Mouse paw edema model: Male mice (strain not specified) were randomly divided into control and treatment groups; SSR-240612 HCl was administered orally (3, 10 mg/kg) or intraperitoneally (0.3, 1 mg/kg) at a predetermined time (not specified) before subcutaneous injection of des-Arg(9)-BK (dose not specified) into the paw; paw edema volume was measured at multiple time points post-BK injection (method not specified); vehicle-treated mice served as controls (n=number not specified) [1]
\n2. Mouse ear edema model: Male mice (strain not specified) received oral SSR-240612 HCl (0.3, 3, 30 mg/kg) or vehicle; capsaicin (dose not specified) was applied topically to the ear to induce edema; ear edema was quantified (method not specified) at specified time points post-capsaicin application (n=number not specified) [1]
\n3. Rat intestinal injury model (splanchnic artery occlusion/reperfusion): Male Sprague-Dawley rats were subjected to splanchnic artery occlusion for a predetermined duration (not specified), followed by reperfusion; SSR-240612 HCl (0.3 mg/kg) was administered intravenously before or during occlusion (timing not specified); control rats received vehicle; after reperfusion, intestinal tissues were harvested to assess tissue destruction (histological scoring) and neutrophil accumulation (method not specified) (n=number not specified) [1]
\n4. Rat UV-induced hyperalgesia model: Male Sprague-Dawley rats were exposed to UV irradiation (dose/duration not specified) to induce thermal hyperalgesia; SSR-240612 HCl (1, 3 mg/kg) was administered orally; thermal hyperalgesia was evaluated using a paw withdrawal test (method not specified) at multiple time points post-administration (n=number not specified) [1]
\n5. Rat formalin-induced nociception model: Male Sprague-Dawley rats received intraplantar injection of formalin (dose not specified) to induce nociceptive responses; SSR-240612 HCl (10, 30 mg/kg) was administered orally before formalin injection (timing not specified); the late phase of nociceptive response (licking/biting behavior) was recorded and quantified over a specified period (n=number not specified) [1]
\n6. Rat sciatic nerve constriction model (neuropathic pain): Male Sprague-Dawley rats underwent unilateral sciatic nerve constriction to induce neuropathic thermal pain; SSR-240612 HCl (20, 30 mg/kg) was administered orally (frequency/timing not specified); thermal pain sensitivity was assessed using a paw withdrawal test (method not specified) at regular intervals (n=number not specified) [1]
\n7. Rat insulin resistance/allodynia model: Male Sprague-Dawley rats were fed with 10% D-glucose in drinking water for 12 weeks to induce insulin resistance, tactile allodynia and cold allodynia; age-matched control rats received regular drinking water; SSR-240612 HCl (0.3, 3, 10, 30 mg/kg) was administered orally to glucose-fed and control rats; tactile allodynia was measured as paw-withdrawal threshold (in grams) using a von Frey filament test, and cold allodynia was assessed as paw-withdrawal response frequency to cold stimulation (method not specified) at multiple time points (0, 1, 3, 6, 24 h) post-administration; plasma glucose/insulin levels, HOMA index and aortic superoxide anion production were measured in glucose-fed rats treated with 10 mg/kg SSR-240612 HCl (n=number not specified, data presented as means±s.e.mean) [2]

[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.

1. SSR-240612 HCl is a novel, potent, orally effective non-peptide bradykinin B1 receptor antagonist with a selectivity for B1 receptors that is 500 to 1000 times higher than that for B2 receptors [1] 2. SSR-240612 HCl antagonizes by directly inhibiting B1 receptor binding and downstream signal transduction (formation of inositol monophosphate), as well as inhibiting B1 receptor-mediated tissue contraction [1] 3. SSR-240612 HCl has anti-inflammatory, tissue-protective, and analgesic effects in vivo, including inhibition of edema, intestinal ischemia/reperfusion injury, thermal hyperalgesia, formalin-induced nociceptive and neuropathic pain [1] 4. SSR-240612 HCl It can reverse tactile and cold-sensory abnormal pain in a rat model of insulin resistance without affecting plasma glucose/insulin levels, insulin resistance (HOMA index) or vascular oxidative stress (aortic superoxide production), indicating that its action is mediated by direct inhibition of B1 receptors in the spinal cord/sensory nerves rather than by regulating metabolic/vascular pathways [2].
5. SSR-240612 HCl Due to its high oral bioavailability and selective B1 receptor antagonism, it is expected to be a candidate drug for the treatment of insulin resistance/diabetes-related sensory polyneuropathy [2].

C42H53CLN4O7S

793.42

792.332

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

464930-42-5

465539-70-2

44235958

White to off-white solid powder

9.395

3

9

14

55

1330

4

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

GLHHFOSVBQQNAW-GDYXXZBVSA-N

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

(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

2934.99.9001

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.

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

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.

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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.

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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.

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