Deucravacitinib (BMS-986165)

Tyk2 JH2 (IC50 = 0.2 nM); JAK1 JH2 (IC50 = 1 nM)

Small molecule JAK inhibitors have emerged as a major therapeutic advancement in treating autoimmune diseases. The discovery of isoform selective JAK inhibitors that traditionally target the catalytically active site of this kinase family has been a formidable challenge. Our strategy to achieve high selectivity for TYK2 relies on targeting the TYK2 pseudokinase (JH2) domain. Herein we report the late stage optimization efforts including a structure-guided design and water displacement strategy that led to the discovery of BMS-986165 (11) as a high affinity JH2 ligand and potent allosteric inhibitor of TYK2. In addition to unprecedented JAK isoform and kinome selectivity, 11 shows excellent pharmacokinetic properties with minimal profiling liabilities and is efficacious in several murine models of autoimmune disease. On the basis of these findings, 11 appears differentiated from all other reported JAK inhibitors and has been advanced as the first pseudokinase-directed therapeutic in clinical development as an oral treatment for autoimmune diseases [1].
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1]. Potential benefits of compounds with deuteration: Longer half-life in living things. Deuterated compounds might be able to increase the compound's pharmacokinetic properties, or in vivo half-life. This can facilitate administration and enhance the compound's safety, effectiveness, and tolerance. Boost oral bioavailability, second. Greater amounts of the unmetabolized medicine are able to reach their target of action because deuterated substances lessen the amount of undesired metabolism (first-pass metabolism) in the liver and intestinal wall. Better tolerance and activity at low doses are determined by high bioavailability. (3) Enhance the properties of metabolism. Deuterated substances can enhance medication metabolism and lessen the production of hazardous or reactive metabolites. (4) Enhance the security of medications. Deuterated chemicals are harmless and can lessen or eliminate the undesirable side effects of medicinal substances. (5) Preserve the treatment outcome. According to earlier research, deuterated molecules should maintain biological potency and selectivity comparable to hydrogen analogs.

Lupus-like disease is strongly inhibited in NZB/W mice treated with Tyk2-IN-4. Tyk2-IN-4 is safe and overall well-tolerated. There are no serious adverse events and the frequency of non-serious adverse events are similar in the active (75%) and placebo (76%) groups. After oral administration, Tyk2-IN-4 is rapidly absorbed and exhibits an apparent elimination half-life of 8-15 hours[1].

Small molecule JAK inhibitors have emerged as a major therapeutic advancement in treating autoimmune diseases. The discovery of isoform selective JAK inhibitors that traditionally target the catalytically active site of this kinase family has been a formidable challenge. Our strategy to achieve high selectivity for TYK2 relies on targeting the TYK2 pseudokinase (JH2) domain. Herein we report the late stage optimization efforts including a structure-guided design and water displacement strategy that led to the discovery of BMS-986165 (11) as a high affinity JH2 ligand and potent allosteric inhibitor of TYK2 [1].
All biochemical potencies and selectivities were determined using homogeneous time-resolved fluorescence (HTRF) assays where compounds were shown to compete with a fluorescent probe for binding to human recombinant JAK1, JAK2, JAK3, and TYK2 JH1 domain proteins in addition to TYK2 and JAK1 JH2 protein domains. Dose–response curves were generated to determine the concentration required for inhibiting 50% of the HTRF signal (IC50) as derived by nonlinear regression analysis. Cellular potencies and selectivities were determined using stably integrated STAT-dependent luciferase reporter assays in T-cells using IFNα-stimulation for measuring TYK2/JAK1 dependent signaling and IL-23 stimulation for measuring TYK2/JAK1 dependent signaling. JAK2 dependent signaling was measured in TF-1 cells using GM-CSF stimulation. Dose–response curves were generated to determine the concentration required to inhibit 50% of cellular response (IC50) as derived by nonlinear regression analysis. Potencies and selectivities for JAK-dependent signaling were also measured in human and mouse whole blood using specific cytokine stimulations and measuring the phosphorylation of specific STAT proteins by cellular staining and flow cytometry. Experimental details for all assays have been previously reported. All compounds active in biological assays were electronically filtered for structural attributes common to pan assay interference compounds (PAINS) and were found to be negative [1].

IL-23-Induced Acanthosis in Mice
Acanthosis was induced in 6–8-week-old C57BL/6 female mice (19–20 g average weight, Jackson Laboratories) by intradermal injection of dual chain, recombinant human IL-23 into the right ear. IL-23 injections were administered every other day from day 0 through day 9 of the study. Treatment groups consisted of eight mice per group. Compound 11 at 7.5, 15, and 30 mg/kg BID in vehicle (EtOH:TPGS:PEG300, 5:5:90) and vehicle alone dosed BID by oral gavage, with the first dose given the evening before the first IL-23 injection. An anti-IL-23 adnectin (3 mg/kg) and PBS control were administered subcutaneously approximately 1 h prior to the first IL-23 injection and then twice a week thereafter. Ear thickness was measured using a Mitutoyo (no. 2412F) dial caliper and calculated as the percent change in thickness from the baseline measurement taken on day 0 before initial IL-23 injections for each animal. At the end of the study, IL-23-injected ears as well as naïve control ears were collected from four animals per group for histological examination and gene expression analyses. Terminal blood samples collected via the retro-orbital sinus were used for PK determinations. Statistical analyses were performed using Student’s t tests or ANOVA with Dunnett’s post test. At the end of the study, ears were removed and fixed in 10% neutral-buffered formalin for 24–48 h. The fixed ears were then cut longitudinally, and two pieces were parallel embedded to make the paraffin blocks. The paraffin blocks were then sectioned and placed on microscope slides for H&E staining for histological evaluation. Severity of ear inflammation was scored using an objective scoring system based on the following parameters: extent of the lesion, severity of hyperkeratosis, number and size of pustules, height of epidermal hyperplasia (acanthosis, measured in interfollicular epidermis), and the amount of inflammatory infiltrate in the dermis and soft tissue. The latter two parameters, acanthosis and inflammatory infiltrate, were scored independently on a scale from 0 to 4: 0, none; 1, minimal; 2, mild; 3, moderate; 4, marked. The histological changes were blindly evaluated by a pathologist. Statistical analyses was performed using one-way ANOVA with Dunnett’s test for comparison of each treatment versus the vehicle control.

[1]. Wrobleski ST, et al. Highly Selective Inhibition of Tyrosine Kinase 2 (TYK2) for the Treatment of Autoimmune Diseases: Discovery of the Allosteric Inhibitor BMS-986165. J Med Chem. 2019 Jul 18.
[2]. Catlett I, et al. SAT0226 A first-in-human, study of BMS-986165, a selective, potent, allosteric small molecule inhibitor of tyrosine kinase 2. Annals of the Rheumatic Diseases 2017;76:859.

C20H22N8O3

425.4590

425.200316

C, 56.46; H, 5.92; N, 26.34; O, 11.28

1609392-27-9

1609392-28-0 (HCl);1609392-27-9;

Off-white to light yellow solid

1.2

136Ų

O=C(C1=NN=C(NC(C2CC2)=O)C=C1NC3=CC=CC(C4=NN(C)C=N4)=C3OC)NC([2H])([2H])[2H]

BZZKEPGENYLQSC-FIBGUPNXSA-N

InChI=1S/C20H22N8O3/c1-21-20(30)16-14(9-15(25-26-16)24-19(29)11-7-8-11)23-13-6-4-5-12(17(13)31-3)18-22-10-28(2)27-18/h4-6,9-11H,7-8H2,1-3H3,(H,21,30)(H2,23,24,25,29)/i1D3

6-(cyclopropanecarboxamido)-4-((2-methoxy-3-(1-methyl-1H-1,2,4-triazol-3-yl)phenyl)amino)-N-(methyl-d3)pyridazine-3-carboxamide

BMS 986165; Deucravacitinib; BMS-986165; Sotyktu; BMS986165

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 : ~33.33 mg/mL (~78.34 mM)

Solubility in Formulation 1: 3.83 mg/mL (9.00 mM) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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 (5.88 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 3: ≥ 2.5 mg/mL (5.88 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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Solubility in Formulation 4: 10 mg/mL (23.50 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.

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