Tryptase (IC50 <1.7 nM)

Potency versus tryptase and selectivity for the two single diastereomers BMS-363131 (2) and BMS-363130 (22) that comprise the mixture 18a are also shown in Table 2. Each of the diastereomers had an IC50 <1.7 nM against tryptase with selectivity >3,000-fold versus trypsin and >10,000-fold versus plasmin, thrombin, uPA, and tPA. BMS-363131 was selected over BMS-363130 for further evaluation following comparisons of other biological properties of the two compounds [1].

In the ovalbumin-sensitized guinea pig model of lung inflammation, intratracheal dosing of BMS-363131 effected a reduction in inflammatory cell count in the lungs compared to control [1].

[1]. Synthesis of potent and highly selective inhibitors of human tryptase. Bioorg Med Chem Lett. 2002 Nov 4;12(21):3235-8.

Serine proteases, such as trypsinoids, are closely associated with allergic and inflammatory diseases, as well as asthma. This article describes a series of synthetic and structure-activity relationship studies of N1-activated-4-carboxylated azacyclic methyl ethyl ketones, ultimately identifying compound BMS-363131 (2) as a potent human trypsin inhibitor (IC50 <1.7 nM) with extremely high selectivity (>3000-fold) against trypsinoids compared to other related serine proteases, including trypsin. In summary, our structure-activity relationship studies of the N-1 and C-3 positions of the azacyclic methyl ethyl ketone core of compound BMS-262084 (1) showed that the conformationally restricted guanidino group at C-3 and the extended substituent on the piperazine ring at N-1 effectively inhibited trypsinoids and significantly improved their selectivity against other serine proteases, including trypsin. Among these, the most potent and selective compound, BMS-363131 (2), demonstrated therapeutic efficacy in a guinea pig asthma model. [1]

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Trypsin is a serine-like protease and a major protein component of mast cells. Activated mast cells degranulate, releasing trypsin, histamine, chymotrypsin, and other allergic and inflammatory mediators. Trypsin is closely associated with allergic, inflammatory, and autoimmune diseases and is thought to play an important role in asthma-related inflammation, bronchoconstriction, and remodeling. Therefore, inhibition of trypsin may have potential therapeutic benefits for asthma. Our previous study reported that BMS-262084 (1) is a potent trypsin inhibitor (IC50 = 4 nM) with moderate to good selectivity for related serine proteases but no selectivity for trypsin. In an ovalbumin-sensitized guinea pig model, intratracheal administration of BMS-262084 showed efficacy in preventing allergen-induced bronchoconstriction and protecting the lungs from inflammatory cell infiltration. Given that most of the inhaled drug is swallowed, good selectivity for pepsin and trypsin is crucial. In this paper, we report our work on improving BMS-262084 by exploring a conformationally restricted guanidino group at the C-3 position and a novel substituent at the N-1 position of the azacyclobutanone core. These studies ultimately led to the discovery of BMS-363131 (2), a potent trypsin inhibitor with excellent selectivity for other serine proteases, including trypsin, and improved hydrolytic stability. [1]

C28H40N6O5

540.65

540.306

C, 62.20; H, 7.46; N, 15.54; O, 14.80

384829-65-6

384830-07-3 (HCl);384829-65-6 9;

10347259

Typically exists as solid at room temperature

2.512

3

6

10

39

914

3

O=C1[C@@H]([C@@H](C(=O)O)N1C(N1CCN(C(CCCCCC2C=CC=CC=2)=O)CC1)=O)C[C@@H]1CN(C(=N)N)CCC1

FHCQLMSOTXBNCN-AKFKNWHVSA-N

InChI=1S/C28H40N6O5/c29-27(30)33-13-7-11-21(19-33)18-22-24(26(37)38)34(25(22)36)28(39)32-16-14-31(15-17-32)23(35)12-6-2-5-10-20-8-3-1-4-9-20/h1,3-4,8-9,21-22,24H,2,5-7,10-19H2,(H3,29,30)(H,37,38)/t21-,22-,24+/m1/s1

(2S,3R)-3-[[(3R)-1-carbamimidoylpiperidin-3-yl]methyl]-4-oxo-1-[4-(6-phenylhexanoyl)piperazine-1-carbonyl]azetidine-2-carboxylic acid

BMS363131; BMS 363131; BMS-363,131; 384829-65-6; UNII-82AT6RBC74; 2-Azetidinecarboxylic acid, 3-(((3R)-1-(aminoiminomethyl)-3-piperidinyl)methyl)-4-oxo-1-((4-(1-oxo-6-phenylhexyl)-1-piperazinyl)carbonyl)-, (2S,3R)-; 82AT6RBC74; CHEMBL303437; (2S,3R)-3-[[(3R)-1-carbamimidoylpiperidin-3-yl]methyl]-4-oxo-1-[4-(6-phenylhexanoyl)piperazine-1-carbonyl]azetidine-2-carboxylic acid; SCHEMBL7666458; BMS-363131

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)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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