Pharmaceutical compounds

Abstract

The invention provides a method of inhibiting a TYK2 kinase, which method comprises bringing into contact with the TYK2 kinase an effective TYK2 kinase-inhibiting amount of a compound having the formula (0): ##STR00001##
or a salt or stereoisomer thereof. The invention also provides a novel subset of compounds within formula (0) as well as pharmaceutical compositions containing them and their use in medicine.

Claims

1. A method of treating a disease or condition in a subject in need thereof, wherein the disease is selected from an autoimmune disease, an inflammatory disease or condition, an immunological disease or condition, an allergic disease or disorder, a transplant rejection and Graft-versus host disease, or a disease or condition selected from sepsis and septic shock, wherein the disease or condition is susceptible to TYK2 inhibition, which method comprises inhibiting a TYK2 kinase by administering to the subject an effective TYK2 kinase-inhibiting amount of 2-(2,6-dichloro-phenyl)-5-[4-(morpholine-4-carbonyl)-phenylamino]-oxazole-4-carboxylic acid amide or a salt thereof.

2. The method of claim 1 wherein the disease or condition is an autoimmune disease.

3. A method according to claim 2 wherein the autoimmune disease is psoriasis.

4. A method according to claim 2 wherein the autoimmune disease is multiple sclerosis.

5. A method according to claim 1 wherein the disease or condition is an inflammatory disease or condition.

6. A method according to claim 1 wherein the disease or condition is an immunological disease or condition.

7. A method according to claim 1 wherein the disease or condition is an allergic disease or disorder.

8. A method according to claim 1 wherein the disease or condition is a transplant rejection.

9. A method according to claim 1 wherein the disease or condition is Graft-versus host disease.

10. A method according to claim 1 wherein the disease or condition is selected from sepsis and septic shock.

11. A method according to claim 1 wherein the disease or condition is selected from: (a) skin inflammation due to radiation exposure; (b) asthma; (c) allergic inflammation; (d) chronic inflammation; (e) an inflammatory ophthalmic disease; (f) dry eye syndrome; (g) uveitis; (h) insulin-dependent diabetes; (i) Hashimoto's thyroiditis; (j) Graves' disease; (k) Cushing's disease; (l) Addison's disease (m) chronic active hepatitis; (n) polycystic ovary syndrome; (o) coeliac disease; (p) psoriasis; (q) inflammatory bowel disease; (r) ankylosing spondylitis; (s) rheumatoid arthritis; (t) systemic lupus erythematosus; (u) myasthenia gravis; (v) transplant rejection; and (w) graft-versus-host disease.

12. A method according to claim 1 wherein the disease or condition is rheumatoid arthritis.

13. A method according to claim 1 wherein the disease or condition is inflammatory bowel disease.

14. A method according to claim 1 wherein the subject is a patient who has been screened and has been determined as suffering from a disease or condition which would be susceptible to treatment with a compound having activity against a TYK2 kinase.

15. A method according to claim 2 wherein the subject is a patient who has been screened and has been determined as suffering from a disease or condition which would be susceptible to treatment with a compound having activity against a TYK2 kinase.

16. A method according to claim 3 wherein the subject is a patient who has been screened and has been determined as suffering from a disease or condition which would be susceptible to treatment with a compound having activity against a TYK2 kinase.

17. A method according to claim 4 wherein the subject is a patient who has been screened and has been determined as suffering from a disease or condition which would be susceptible to treatment with a compound having activity against a TYK2 kinase.

18. A method according to claim 5 wherein the subject is a patient who has been screened and has been determined as suffering from a disease or condition which would be susceptible to treatment with a compound having activity against a TYK2 kinase.

19. A method according to claim 6 wherein the subject is a patient who has been screened and has been determined as suffering from a disease or condition which would be susceptible to treatment with a compound having activity against a TYK2 kinase.

20. A method according to claim 11 wherein the subject is a patient who has been screened and has been determined as suffering from a disease or condition which would be susceptible to treatment with a compound having activity against a TYK2 kinase.

Description

EXAMPLES

(1) The invention will now be illustrated, but not limited, by reference to the specific embodiments described in the following examples.

Enzyme Inhibition

(2) Compounds of the invention were assayed for their ability to inhibit TYK2 kinase and other JAK kinases.

(3) Substrates and kinases used in the assays are identified in Table 2 below.

(4) Kinase assays were performed at Reaction Biology Corp., Malvern, Pa., USA, using the following general procedure:

(5) 1) Prepare indicated substrate in freshly prepared Base Reaction Buffer (20 mM Hepes pH 7.5, 10 mM MgCl.sub.2, 1 mM EGTA, 0.02% Brij35, 0.02 mg/ml BSA, 0.1 mM Na.sub.3VO.sub.4, 2 mM DTT, 1% DMSO).

(6) 2) Deliver cofactors (1.5 mM CaCl.sub.2, 16 ug/mL Calmodulin, 2 mM MnCl.sub.2) to the substrate solution above

(7) 3) Deliver indicated kinase into the substrate solution and gently mix

(8) 4) Deliver varying concentrations of test compound in DMSO into the kinase reaction mixture

(9) 5) Deliver .sup.33P-ATP (specific activity 0.01 μCi/μL final) into the reaction mixture to initiate the reaction

(10) 6) Incubate kinase reaction for 120 min at room temperature

(11) 7) Reactions are spotted onto P81 ion exchange filter paper (Whatman #3698-915)

(12) 8) Unbound phosphate is removed by washing filters extensively in 0.75% Phosphoric acid.

(13) 9) .sup.33P signal was determined using Typhoon phosphorimagers (GE Healthcare). After subtraction of background derived from control reactions containing inactive enzyme, IC.sub.50 values were determined using the nonlinear regression function in Prism (Graphpad software).

(14) TABLE-US-00003 TABLE 2 Protein Protein HUGO Genbank Accession Name symbol Substrate Accession # # Clone Expression Tag JAK1 JAK1 pEY NP_002218.2 P23458 aa 866- Baculovirus N- 1154 in Sf21 terminal insect cells GST tag JAK2 JAK2 pEY NP_004963 O60674 aa 809- Baculovirus N- 1132 + in Sf21 terminal g insect cells GST tag JAK3 JAK3 JAK3tide NP_000206 P52333 aa 781- Baculovirus N- 1124 in Sf21 terminal insect cells GST tag TYK2 TYK2 AXLtide NP_003322.2 P29597 Aa Baculovirus N- 833- in Sf21 terminal 1187 insect cells GST tag

(15) Substrates:

(16) AXLtide=[KKSRGDYMTMQIG]

(17) JAK3tide=[Ac-GEEEEYFELVKKKK-NH.sub.2]

(18) pEY=poly Glu-Tyr [Glu:Tyr (4:1), M.W.=5,000−20,000]

(19) The results are shown in Table 3 below.

(20) TABLE-US-00004 TABLE 3 Example Number (and method of In Vitro Enzyme IC.sub.50 (nM) preparation) STRUCTURE TYK2 JAK1 JAK2 JAK3 Example 1 (Example Q-3 in WO2008/139161) 13.5 90.2 234.7 404.8 Example 2 (Example Q-25 in WO2008/139161) 14.6 78.2 146.6 418.8 Example 3 (Example Q-26 in WO2008/139161) 5.3 47.6 95.0 359.0 Example 4 (Example Q-27 in WO2008/139161) 13.8 65.6 109.0 387.2 Example 5 (Example Q-20 in WO2008/139161) 9.2 88.6 112.1 218.9 Example 6 (Example Q-51 in WO2008/139161) 25.0 192.4 297.1 471.6 Example 7 (Example Q-54 in WO2008/139161) 9.8 201.5 261.0 419.3 Example 8 (Example Q-53 in WO2008/139161) 0 12.9 201.0 267.5 408.5 Example 9 (Example U-2 in WO2008/139161) 22.7 75.6 267.4 423.4 Example 10 (Example U-3 in WO2008/139161) 20.5 183.3 311.4 397.2 Example 11 (Example U-4 in WO2008/139161) 15.1 189.6 338.4 387.7 Example 12 (Example U-6 in WO2008/139161) 23.41 168.6 292.4 346.2 Example 13 (Example U-7 in WO2008/139161) 11.2 123.0 181.6 341.5 Example 14 (Example U-12 in WO2008/139161) 9.6 67.22 36.0 125.9 Example 15 (Example U-16 in WO2008/139161) 7.5 41.1 101.3 194.9 Example 16 (Example U-17 in WO2008/139161) 8.4 58.1 118.8 199.1 Example 17 (Example U-21 in WO2008/139161) 13.7 152.8 167.2 99.2 Example 18 (Example U-18 in WO2008/139161) 0 13.8 118.3 191.8 164.6

(21) The data set out in the table above illustrate that compounds of formula (1) are potent inhibitors of TYK2 kinase and show a pronounced selectivity for TYK2 kinase compared to other JAK kinases.

(22) On the basis of their activity against TYK2 kinase, it is envisaged that the compounds of the formula (1) will be useful as therapeutic agents for treating a wide range of inflammatory, immunological and allergic diseases and conditions.

Examples 19 to 33

(23) The compounds of Examples 19 to 33 in Table 4 below are novel compounds and are made using the methods described below or methods analogous thereto. The starting materials and synthetic intermediates used in the methods are shown in Table 5 and the NMR and LCMS properties of the final products are set out in Table 6.

(24) TABLE-US-00005 TABLE 4 Example 19 Example 20 Example 21 Example 22 Example 23 Example 24 Example 25 Example 26 Example 27 0 Example 28 Example 29 Exampe 30 Example 31 Example 32 Example 33

(25) TABLE-US-00006 TABLE 5 Synthetic Intermediates and Starting Materials Compound Structure Source or method of preparation I-1  Prepared as described in steps a and b from Example F-1 in WO2008/139161, using 2,6- dichlorobenzoyl chloride in step a I-2  Prepared as described in steps a and b from Example F-1 in WO2008/139161, using 2-chloro-6- fluorobenzoyl chloride in step a I-3  Prepared as described in steps a and b from Example F-1 in WO2008/139161, using 2,4,6- trifluorobenzoyl chloride in step a I-4  Prepared as described in steps a and b from Example F-1 in WO2008/139161, using 2,5- difluorobenzoyl chloride in step a I-5  0 Example F-1 of WO2008/139161 I-6  Commercially available I-7  Commercially available I-8  Commercially available I-9  Commercially available I-10   (Compound (13) in General Method B) Commercially available I-11 Commercially available I-12 Commercially available I-13 Commercially available

General Method a

Step a—Preparation of Intermediate Compound (12)

(26) ##STR00089##

(27) In the reaction scheme, the group R.sup.1′ in formulae (11) and (12) is either a group R.sup.1 as defined herein or a protected form of the group R.sup.1.

(28) A solution of palladium acetate (0.025 mmol) and (±)-2,2″-bis(diphenylphosphino)-1,1″-binaphthalene (0.024 mmol) in DMF (7.1 mL) is stirred at room temperature for 3 minutes. Then compound (10) (0.35 mmol), compound (11) (1.40 mmol) and potassium phosphate tribasic (0.70 mmol) are added and the mixture heated in the microwave for 3 minutes at 180° C. The reaction is diluted with EtOAc and washed with water. The organic phase is passed through a MP-SH resin cartridge, dried over MgSO.sub.4 and the solvent removed in vacuo. The residue is purified by silica gel column chromatography using a gradient 10-100% EtOAc in hexanes to afford Compound (12), the identity of which can be confirmed by .sup.1H NMR (DMSO) and LCMS.

Step b—Preparation of Compound (2)

(29) ##STR00090##

(30) A solution of Compound (12) (0.09 mmol) in concentrated sulfuric acid (1.7 mL) is stirred at room temperature for 1.5 hours. The solution is neutralised by pouring into saturated sodium bicarbonate solution. The aqueous phase is extracted with EtOAc. The combined organic phase is dried over MgSO.sub.4 and the solvent is removed in vacuo to afford Compound (2), the identity of which can be confirmed by .sup.1H NMR (DMSO) and LCMS.

General Method B

Step a—Preparation of Intermediate Compound (14)

(31) ##STR00091##

(32) Compound (10) is reacted with compound (13) under the conditions set out in step a of General Method A to give Compound (14).

Step b—Preparation of Intermediate Compound (15)

(33) ##STR00092##

(34) Compound (14) is hydrolysed using lithium hydroxide to give the carboxylic acid Compound (15).

(35) Alternatively, Compound (15) can be prepared by the method of step a of Example U-1 of WO2008/139161 or methods analogous thereto.

Step c—Preparation of Intermediate Compound (16)

(36) ##STR00093##

(37) To a solution of Compound (15) (0.059 mmol), O-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophosphate (0.059 mmol), and diisopropylethylamine (0.117 mmol) in N,N-dimethylformamide (2 mL) is added an amine of formula HNR.sup.xR.sup.y (0.059 mmol) and the reaction mixture is stirred at room temperature for 16 hours. The reaction mixture is then diluted with EtOAc washed with 1M HCl, water and brine. The organic phase is dried over MgSO.sub.4 and the solvent removed in vacuo. The residue is purified by preparative HPLC to afford Compound (16), the identity of which can be confirmed by LCMS.

Step d—Preparation of Compound (17)

(38) ##STR00094##

(39) A solution of Compound (16) (0.022 mmol) in concentrated sulfuric acid (0.5 mL) is stirred at room temperature for 1.5 hours. The solution is neutralised by pouring into saturated sodium bicarbonate solution. The aqueous phase is then basified to pH14 using 5M NaOH and extracted with EtOAc. The combined organic phase is dried over MgSO.sub.4 and the solvent removed in vacuo to afford Compound (17), the identity of which can be confirmed by .sup.1H NMR (DMSO) and LCMS. Compounds (17) containing a basic nitrogen, such as those prepared from I-11 or I-12 which bear an acid-sensitive nitrogen protecting group, are concomitantly deprotected during the final acid-mediated reaction step.

(40) General Method B can be used to make compounds wherein NR.sup.xR.sup.y forms a cyclic amine such as a morpholino, piperazino or piperidino group or compounds wherein R.sup.x is hydrogen or a substituent and R.sup.y is hydrogen or a substituent.

(41) TABLE-US-00007 TABLE 6 Ex. No. Name Synthetic method .sup.1H NMR LCMS 19 2-(2,6-Dichloro-phenyl)-5- General Method A using (DMSO) δ 9.86 (1H, s), 7.83 (2H, d), m/z (ES+) 426 (4-methanesulfonyl- intermediates I-1 and I-6. 7.73-7.65 (4H, m), 7.48 (3H, d), 3.14 phenylamino)-oxazole-4- (3H, s) carboxylic acid amide 20 2-(2-Chloro-6-fluoro- General Method A using (DMSO) δ 9.86 (1H, s), 7.83 (2H, d), m/z (ES+) 410 phenyl)-5-(4- intermediates I-2 and I-6. 7.64 (1H, q), 7.58-7.48 (6H, m), 3.16 methanesulfonyl- (3H, s) phenylamino)-oxazole-4- carboxylic acid amide 21 5-(4-Methanesulfonyl- General Method A using (DMSO) δ 9.81 (1H, s), 7.81 (2H, d), m/z (ES+) 412 phenylamino)-2-(2,4,6- intermediates I-3 and I-6. 7.54 (2H, d), 7.46-7.44 (4H, m), 3.15 trifluoro-phenyl)-oxazole-4- (3H, s) carboxylic acid amide 22 2-(2,5-Difluoro-phenyl)-5-(4- General Method A using (DMSO) δ 9.84 (1H, s), 7.85 (2H, d), m/z (ES+) 394 methanesulfonyl- intermediates I-4 and I-6. 7.78 (1H, br. s), 7.63 (2H, d), 7.51- phenylamino)-oxazole-4- 7.45 (4H, m), 3.19 (3H, s) carboxylic acid amide 23 (S) 2-(2-Chloro-6-fluoro- General Method B using phenyl)-5-[4-(piperidin-3- intermediates I-2, I-10 and I-11 ylcarbamoyl)-phenylamino]- oxazole-4-carboxylic acid amide 24 (R) 2-(2-Chloro-6-fluoro- General Method B using (MeOD) δ 7.87 (2H, d), 7.62-7.59 m/z (ES+) 458 phenyl)-5-[4-(piperidin-3- intermediates I-2, I-10 and I-12 (1H, m), 7.49-7.46 (3H, m), 7.33 (1H, ylcarbamoyl)-phenylamino]- t), 4.29-4.24 (1H, m), 3.53 (1H, dd), oxazole-4-carboxylic acid 3.39-3.35 (1H, m), 3.02-2.92 (2H, amide m), 2.13-2.07 (2H, m), 1.89-1.74 (2H, m) 25 2-(2-Chloro-6-fluoro- General Method B using (DMSO) δ 9.50 (1H, s), 7.64 (1H, m/z (ES+) 445 phenyl)-5-[4-(morpholine-4- intermediates I-2, I-10 and dd), 7.54 (1H, d), 7.47-7.45 (2H, m), carbonyl)-phenylamino]- morpholine 7.39-7.34 (5H, m), 3.55 (4H, br. m), oxazole-4-carboxylic acid 3.36 (4H, br. m) amide 26 2-(2-Chloro-6-fluoro- General Method B using (MeOD) δ 7.87 (2H, d), 7.62-7.58 m/z (ES+) 472 phenyl)-5-[4-(1-methyl- intermediates I-2, I-10 and I-13 (1H, m), 7.51-7.47 (3H, m), 7.34 (1H, piperidin-4-ylcarbamoyl)- t), 4.16 (1H, m), 3.59 (2H, m), 3.20 phenylamino]-oxazole-4- (2H, m), 2.92 (3H, s), 2.24 (2H, m), carboxylic acid amide 1.92 (2H, m) 27 (S) 2-(2,6-Dichloro-phenyl)- General Method B using (MeOD) δ 7.82 (2H, d), 7.59-7.52 m/z (ES+) 474 5-[4-(piperidin-3- intermediates I-1, I-10 and I-11 (3H, m), 7.40 (2H, d), 4.23-4.18 (1H, ylcarbamoyl)-phenylamino]- m), 3.49 (1H, dd), 3.34-3.29 (1H, m), oxazole-4-carboxylic acid 2.97-2.86 (2H, m), 2.09-2.03 (2H, m) amide 1.84-1.69 (2H, m) 28 (R) 2-(2,6-Dichloro-phenyl)- General Method B using (MeOD) δ 7.86 (2H, d), 7.64-7.60 m/z (ES+) 474 5-[4-(piperidin-3- intermediates I-1, I-10 and I-12 (3H, m), 7.44 (2H, d), 4.25-4.23 (1H, ylcarbamoyl)-phenylamino]- m), 3.52 (1H, dd), 3.38-3.33 (1H, m), oxazole-4-carboxylic acid 3.02-2.90 (2H, m), 2.11-2.07 (2H, m) amide 1.88-1.74 (2H, m) 29 2-(2,6-Dichloro-phenyl)-5- General Method B using [4-(morpholine-4-carbonyl)- intermediates I-1, I-10 and phenylamino]-oxazole-4- morpholine carboxylic acid amide 30 2-(2,6-Dichloro-phenyl)-5- General Method B using (MeOD) δ 7.85 (2H, d), 7.62-7.59 m/z (ES+) 490 [4-(1-methyl-piperidin-4- intermediates I-1, I-10 and I-13 (3H, m), 7.43 (2H, d), 4.15 (1H, m), ylcarbamoyl)-phenylamino]- 3.58 (2H, m), 3.19 (2H, m), 2.91 (3H, oxazole-4-carboxylic acid s), 2.22 (2H, m), 1.93 (2H, m) amide 31 2-(2,6-Difluoro-phenyl)-5-(4- General Method A using (DMSO) δ 9.82 (1H, s), 7.74 (2H, d), m/z (ES+) 430 ethanesulfonyl- intermediates I-5 and I-7. 7.62-7.60 (1H, m), 7.54 (2H, d), 7.44 (M + Na.sup.+) phenylamino)-oxazole-4- (2H, br s), 7.33-7.29 (2H, m), 3.20 carboxylic acid amide (2H, q), 1.06 (3H, t) 32 2-(2,6-Difluoro-phenyl)-5-(4- General Method A using (DMSO) δ 10.07 (1H, s), 7.73-7.67 m/z (ES+) 434 methanesulfonyl- intermediates I-5 and I-6. (2H, m), 7.52-7.45 (3H, m), 7.40- (M + Na.sup.+) phenylamino)-oxazole-4- 7.34 (3H, m), 3.27 (3H, s) carboxylic acid amide 33 2-(2,6-Difluoro-phenyl)-5[4- General Method A using (DMSO) δ 9.83 (1H, s), 7.71 (2H, d), m/z (ES+) 444 propane-2-sulfonyl)- intermediates I-5 and I-8. 7.63-7.60 (1H, m), 7.54 (2H, d), 7.44 (M + Na.sup.+) phenylamino]-oxazole-4- (2H, br s), 7.33-7.29 (2H, m), 3.35 carboxylic acid amide (1H, m), 1.11 (6H, d)

Example 34

Enzyme Inhibitory Activities of Novel Compounds of Formula (2)

(42) Novel compounds of formula (2) were tested in the TYK2 kinase inhibition assay and the other JAK kinase inhibition assays described above. The results are shown in Table 7 below.

(43) TABLE-US-00008 TABLE 7 Compound of In Vitro Enzyme IC.sub.50 (nM) Example No. TYK2 JAK1 JAK2 JAK3 19 2.3 21.9 87.7 214 20 2.7 28.7 72.6 165 21 68.3 241 412 2180 22 183 843 663 5500

Example 35

Pharmaceutical Formulations

(i) Tablet Formulation

(44) A tablet composition containing a compound of the formula (0), (1) or (2) is prepared by mixing 50 mg of the compound with 197 mg of lactose (BP) as diluent, and 3 mg magnesium stearate as a lubricant and compressing to form a tablet in a known manner.

(ii) Capsule Formulation

(45) A capsule formulation is prepared by mixing 100 mg of a compound of the formula (0), (1) or (2) with 100 mg lactose and filling the resulting mixture into standard opaque hard gelatin capsules.

(iii) Injectable Formulation I

(46) A parenteral composition for administration by injection can be prepared by dissolving a compound of the formula (0), (1) or (2) (e.g. in a salt form) in water containing 10% propylene glycol to give a concentration of active compound of 1.5% by weight. The solution is then sterilised by filtration, filled into an ampoule and sealed.

(iv) Injectable Formulation II

(47) A parenteral composition for injection is prepared by dissolving in water a compound of the formula (0), (1) or (2) (e.g. in salt form) (2 mg/mL) and mannitol (50 mg/mL), sterile filtering the solution and filling into sealable 1 mL vials or ampoules.

(iv) Sub-Cutaneous Injection Formulation

(48) A composition for sub-cutaneous administration is prepared by mixing a compound of the formula (0), (1) or (2) with pharmaceutical grade corn oil to give a concentration of 5 mg/mL. The composition is sterilised and filled into a suitable container.

Equivalents

(49) The foregoing examples are presented for the purpose of illustrating the invention and should not be construed as imposing any limitation on the scope of the invention. It will readily be apparent that numerous modifications and alterations may be made to the specific embodiments of the invention described above and illustrated in the examples without departing from the principles underlying the invention. All such modifications and alterations are intended to be embraced by this application.