11β-HSD1/11β-hydroxysteroid dehydrogenase type 1

In addition to its possible clinical applications in the treatment of type II diabetes, 11β-HSD1 inhibition may also be beneficial in the prevention of atherosclerosis and cognitive decline. In HEK and 3T3L1 cells, BMS-816336 (6n-2) inhibits the 11β-HSD1 enzyme with IC50 values of 37.3 and 28.6 nM, respectively [1].

BMS-816336 is a possible novel medication for metabolic syndrome, type 2 diabetes, and other glucocorticoid-regulated human disorders. In DIO mice (1, 3, 10, 30, 100 mg/kg, 120 minutes) and cynomolgus monkeys (ED50=0.12 mg/kg), BMS-816336 (6n-2) demonstrates strong acute pharmacodynamic effects. Its oral bioavailability ranges from 20% to 72% in preclinical species. Its expected pharmacokinetic properties in humans include a short half-life and a peak to trough ratio [1].

11β-HSD1 SPA Enzyme Assay[1]
11β-HSD1 was assayed by scintillation proximity assay (SPA) in a 384-well PerkinElmer white plate. The dose response of the compounds was determined using 11 half-log dilutions of compound in DMSO in duplicate. To each well, 0.5 μL of compound dilution in DMSO were added. Then 15 μL of assay buffer (for blanks) or 15 μL of human microsomes in assay buffer were added next and the plates were incubated for 10 min at room temperature. The final microsomal protein concentration was 1.1 μg/assay. Duplicates were in the same plate one row below the other. Then 10 μL of 3H-cortisone (final concentration 40 nM) was added to each well and the plate was spun down to mix and bring down the contents to the bottom of the wells. The plates were incubated at room temperature with gentle shaking for 4 h. The reaction was stopped with addition of 10 μL of 10 mM carbenoxolone. Then 0.5 mg of yttrium silicate SPA beads coupled to anticortisol antibody in 20 μL were added to all the wells of plate, which were spun down once more and incubated at room temperature overnight. The plate was read in a TopCount (1 min/well). Data were uploaded automatically to Tool Set, a Lead Evaluation informatics program for data capture and calculation. Graphs were generated with the Curve Master program.

Animal/Disease Models: non-fasting diet-induced obese male mice [1]
Doses: 1, 3, 10, 30, 100 mg/kg
Route of Administration: po (po (oral gavage)) 120 minutes
Experimental Results: ED50=8.6 mg/kg, plasma EC50 is 0.85 μM model [1].

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In Vivo Pharmacodynamic Assessment in Mice[1]
Nonfasting diet-induced obese male mice were weighed and separated into groups (n = 6) such that body weights were not statistically different from each other. Animals were bled via the tail for a −60 min time sample and then were dosed orally with vehicle or drug. The vehicle was composed of 0.5% Methocel, 0.1% Tween 80 in water. At 60 min after dosing, mice were bled again via the tail and dosed orally with DHC @ 10 mg/kg. All animals were subsequently bled at 30, 60, and 120 min post DHC dosing. Plasma was isolated for analysis of corticosterone using a commercially available enzyme immunosorbent assay. Drug levels were also measured in the terminal bleed samples.

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In Vivo Pharmacodynamic Assessment in Cynomolgus Monkey[1]
The experimental design for the cynomolgus monkey pharmacodynamics study is very similar to the mouse model described above. The test compound was administered at a predetermined time point prior to initiation of the experiment based upon the pharmacokinetics of the compound. The monkeys were then given a dose of substrate, and blood samples were removed at various time points thereafter. A notable difference from the mouse protocol was that even though the natural substrate for 11β-HSD-1 in primates was cortisone, the natural rodent substrate 11-dehydrocorticosterone (DHC) was used instead.

Following intravenous injection, BMS-816336 (6n-2) exhibited high total plasma clearance (CLTp) in rats, moderate clearance in mice and monkeys, and low clearance in dogs. Estimated apparent elimination half-lives were 2 hours in mice, 3 hours in rats, 7 hours in dogs, and 6 hours in monkeys. BMS-816336 (6n-2) was extravascularly distributed in all tested animals, with steady-state volumes of distribution (Vss) of 2.0, 0.5, 3.0, and 4.2 L/kg in mice, rats, dogs, and monkeys, respectively. In mice, the tissue/plasma concentration ratio was approximately 0.15 on average in adipose tissue, but concentration-dependent in the liver (1 to 56). Compound 6n-2 was considered unable to cross the blood-brain barrier in mice because its brain tissue concentrations were below the limit of quantification, and the brain/plasma ratio was less than 0.1 at all study time points. The absolute oral bioavailability of 6n-2 administered in homogeneous suspension was estimated to be 56% in mice, 20% in rats, 72% in dogs, and 57% in monkeys. Overall, 6n-2 exhibited favorable in vivo pharmacokinetic characteristics with a higher peak-to-trough ratio, a shorter mean residence time (MTR), and a higher clearance rate, which significantly distinguished it from our two previous clinical compounds. The pharmacokinetic properties of 6n-2 appear to be suitable for once-daily administration, with “transient” inhibition of the enzyme over 24 hours. Therefore, we are eager to test the activation of the hypothalamic-pituitary-adrenal axis (HPA) of this compound in cynomolgus monkeys, a study that will be discussed later. [1]

[1]. Discovery of Clinical Candidate 2-((2S,6S)-2-Phenyl-6-hydroxyadamantan-2-yl)-1-(3'-hydroxyazetidin-1-yl)ethanone [BMS-816336] an Orally Active Novel Selective 11β-Hydroxysteroid Dehydrogenase Type 1 Inhibitor. J Med Chem. 2017 Jun 22;60(12):4932-4948.

BMS-816336 (6n-2) is a hydroxylated adamantyl acetamide that has been identified as a novel and potent inhibitor of human 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) (IC50 3.0 nM) with a selectivity of over 10,000-fold for human 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2). 6n-2 has demonstrated significant acute pharmacodynamic effects in cynomolgus monkeys (ED50 0.12 mg/kg) and diet-induced obese (DIO) mice. It exhibits good oral bioavailability (20% to 72% in preclinical animal models) and is predicted to have a high peak-to-trough ratio and a short half-life in humans. The ADME properties of this drug meet our selection criteria for once-daily dosing, which aims to effectively inhibit the 11β-HSD1 enzyme within the first 12 hours after administration, followed by an "inhibition phase" to reduce the risk of activation of the hypothalamic-pituitary-adrenal (HPA) axis. Phase I clinical studies have shown that 6n-2 is well-tolerated and has the potential to become a new therapy for type 2 diabetes, metabolic syndrome and other glucocorticoid-regulated human diseases. [1] In summary, 6n-2 is a highly potent and selective human 11β-HSD1 inhibitor with excellent water solubility, good in vitro and in vivo safety and acceptable pharmacokinetic properties. Structure-based drug design led to the introduction of a 6-hydroxyl group on the adamantane ring, which is the best substituent for balancing activity and metabolic stability. Despite structural similarities, we observed that the binding orientation of compounds 4k and 6n-2 was reversed in the co-crystal structure with human 11β-HSD1 enzyme. The pharmacokinetic profile of 6n-2 was chosen to enable partial (e.g., 12-hour) enzyme inhibition in vivo after once-daily administration, allowing enzyme activity to "pause" for a period of time, which may help alleviate the activation of the hypothalamic-pituitary-adrenal (HPA) axis that is typically caused by other inhibitors of this enzyme. While we suspect that the pharmacokinetic profile of 6n-2 may alter its ability to activate the HPA axis, it is noteworthy that the concentration of 6n-2 in the brain is not high (the brain/plasma concentration ratio in preclinical animal models was approximately 0.05), which may also be one of the reasons why significant HPA axis activation was not observed in cynomolgus monkeys. Based on acceptable pre-application safety and toxicity data, 6n-2 has entered a Phase I clinical trial and has shown good tolerability at the highest test dose of 900 mg. Data from this study will be published in due course. In addition to the treatment of type II diabetes and metabolic syndrome, 11β-HSD1 inhibitors may have other potential clinical applications, such as anti-atherosclerosis and cognitive protection. These areas need further exploration. [1]

C21H27NO3

341.443986177444

341.199

C, 73.87; H, 7.97; N, 4.10; O, 14.06

1009583-20-3

(Rac)-BMS-816336;(R)-BMS-816336;1009583-83-8; 1009583-20-3 1009365-98-3

59336911

White to off-white solid powder

1.7

2

3

3

25

505

0

OC1C2CC3CC1CC(C2)C3(C1C=CC=CC=1)CC(N1CC(C1)O)=O

OAAZMUGLOXGVNH-MEMOLBONSA-N, OAAZMUGLOXGVNH-CCVYDFRESA-N

InChI=1S/C21H27NO3/c23-18-11-22(12-18)19(24)10-21(15-4-2-1-3-5-15)16-6-13-7-17(21)9-14(8-16)20(13)25/h1-5,13-14,16-18,20,23,25H,6-12H2/t13-,14?,16-,17+,20-,21-/m1/s1

2-((1R,2R,3S,5R,6R)-6-hydroxy-2-phenyladamantan-2-yl)-1-(3-hydroxyazetidin-1-yl)ethan-1-one

BMS-816336; BMS 816336; 1009583-20-3; (Rac)-BMS-816336; (R)-BMS-816336; HLF8J24L87; 1009583-83-8; 1-(3-hydroxyazetidin-1-yl)-2-(6-hydroxy-2-phenyl-2-adamantyl)ethanone; Ethanone, 1-(3-hydroxy-1-azetidinyl)-2-(6-hydroxy-2-phenyltricyclo(3.3.1.13,7)dec-2-yl)-; BMS816336.

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 : ~300 mg/mL (~878.63 mM)

Solubility in Formulation 1: ≥ 7.5 mg/mL (21.97 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 75.0 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: ≥ 7.5 mg/mL (21.97 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 75.0 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: ≥ 7.5 mg/mL (21.97 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 75.0 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.)