ROR gamma (RORγ) modulators

Abstract

The present application relates to compounds according to (Formula IA) or (Formula IB): ##STR00001## or a pharmaceutically acceptable salt thereof. The compounds can be used as inhibitors of RORγ and are useful for the treatment of RORγ mediated diseases.

Claims

1. A pharmaceutical composition in unit dosage form for oral or parenteral administration, said pharmaceutical composition comprising: a compound having the absolute configuration corresponding to Formula IA 2-{4-[(cyclopropylmethyl)sulfonyl]phenyl}-N-{4-[(1R)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxyethyl]phenyl}acetamide: ##STR00025## or a pharmaceutically acceptable salt thereof; and, one or more pharmaceutically acceptable excipients.

2. A process of preparing a compound of Formula (IA) comprising: i) obtaining 2-(4-bromophenyl)-1,1,1,3,3-pentafluoro-propan-2-ol (IV) from 1-(4-bromophenyl)-2,2-fluoro-ethanone (V) by trifluoromethylation using trifluoromethyltrimethylsilane (TMSCF.sub.3) in the presence of tetra-N-butylammonium fluoride (TBAF): ##STR00026## ii) obtaining a compound of Formula (II) by amine substitution of the bromine atom of 2-(4-bromophenyl)-1,1,1,3,3-pentafluoro-propan-2-ol (IV): ##STR00027## wherein said substitution is performed using lithium bis(trimethylsilyl)amide), 2-(dicyclohexylphosphino)biphenyl and tris(dibenzylidene acetone)dipalladium; iii) reacting said amine of formula (II) with a compound of Formula (III): ##STR00028## to give the compound of Formula (I); ##STR00029## wherein said reaction is performed in the presence of 1-propanephosphonic anhydride and N,N-diisopropylethylamine; and, iv) obtaining the compound of Formula (IA) by chiral separation ##STR00030##

3. A method for treatment of at least one condition selected from the group consisting of multiple sclerosis, inflammatory bowel disease, Crohn's disease, psoriasis, rheumatoid arthritis, asthma, osteoarthritis, Kawasaki disease, Hashimoto's thyroiditis and mucosal leishmaniasis, said method comprising administering to a person in need of the treatment a pharmaceutical composition comprising one or more pharmaceutically acceptable excipients and from 0.0001-100 mg per kilogram of bodyweight of a compound having the absolute configuration corresponding to Formula IA or a pharmaceutically acceptable salt thereof, wherein Formula IA is 2-{4-[(cyclopropylmethyl)sulfonyl]phenyl}-N-{4-[(1R)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxyethyl]phenyl}acetamide: ##STR00031##

4. The method according to claim 3, wherein the at least one condition is selected from the group consisting of rheumatoid arthritis, psoriasis, inflammatory bowel disease, Crohn's disease and multiple sclerosis.

5. The method according to claim 3, wherein the at least one condition is selected from osteoarthritis and asthma.

6. The method according to claim 3, wherein the at least one condition is mucosal leishmaniasis.

Description

EXAMPLES

(1) The present application will be explained with reference to examples. However, the scope of the present application is not limited to the following examples.

(2) The compounds of the application can be readily prepared according to the following reaction scheme, or modifications thereof, using readily available starting materials, reagents or previously described intermediates and conventional synthesis procedures.

(3) ##STR00010##

(4) The abbreviations used in this scheme and these experimental details are listed below and additional ones should be considered known to a person skilled in the art of synthetic chemistry.

(5) Abbreviations used herein are as follow: TMSCF.sub.3: Trifluoromethyltrimethylsilane; TBAF: Tetra-N-butylammonium fluoride; TMS: Trimethylsilyl; LiHMDS: Lithium bis(trimethylsilyl)amide; Pd.sub.2(dba).sub.3: Tris(dibenzylideneacetone)dipalladium(0); T3P: 1-Propanephosphonic anhydride; DIPEA: Diisopropylethylamine, N-ethyl-N-isopropyl-propan-2-amine; RT: room temperature; DMF: Dimethylformamide; CH.sub.2Cl.sub.2, DCM: dichloromethane; THF: Tetrahydrofuran; Et.sub.2O: Diethyl ether; DMSO: Dimethylsulfoxide; EtOH: Ethanol; TLC: Thin Layer Chromatography; EtOAc: ethyl acetate; ACN, CH.sub.3CN: acetonitrile; MeOH: methanol; TFA: Trifluoroacetic acid; HCl: Hydrochloric acid; Et.sub.3N, TEA: triethylamine; NaCl: sodium chloride; NaHCO.sub.3: sodium bicarbonate; H.sub.2O: water; MgSO.sub.4: magnesium sulfate; Na.sub.2SO.sub.4: sodium sulfate.

(6) Chemical names are preferred IUPAC names, generated using Accelrys Draw 4.1.

(7) If a chemical compound is referred to using both a chemical structure and a chemical name, and an ambiguity exists between the structure and the name, the structure predominates.

Example 1

(8) 2-[4-(cyclopropylmethylsulfonyl)phenyl]-N-[4-[1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide.

(9) ##STR00011##

(10) Step 1:

(11) ##STR00012##

(12) To a solution of 1-(4-bromophenyl)-2,2-difluoro-ethanone (10.5 g) in 100 mL THF at 0° C. under argon was added TMSCF.sub.3 (12.7 g) followed by addition of a 1 M solution of TBAF in THF (90 mL) over 45 minutes. The reaction mixture was stirred for 1 hour at RT, then diluted with Et.sub.2O (200 mL), washed with water (2×200 mL), brine (100 mL), dried and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel, using 0% to 20% EtOAc in cyclohexane as the eluent to give 11.5 g (84%) of 2-(4-bromophenyl)-1,1,1,3,3-pentafluoro-propan-2-ol as a yellow oil.

(13) MS(ES.sup.+) m/z 302.9/304.9 [M+H].sup.+.

(14) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 6.70 (t, J=53.1 Hz, 2 H), 7.60 (d, J=9.2 Hz, 2 H), 7.69 (d, J=9.2 Hz, 2 H), 7.92 (s, 1 H).

(15) Step 2:

(16) ##STR00013##

(17) To a solution of 2-(4-bromophenyl)-1,1,1,3,3-pentafluoro-propan-2-ol (11.4 g) in THF (100 mL) under argon was added a 1 M solution of LiHMDS in THF (112 mL), 2-(dicyclohexylphosphino)biphenyl (1.6g) followed by addition of Pd.sub.2(dba).sub.3 (2.16g). The reaction mixture was stirred under reflux for 1 hour, then cooled to 0° C., and a 12N aqueous solution of HCl (15 mL) was added dropwise. After stirring for 1 hour at RT, the reaction mixture was poured onto a saturated aqueous solution of NaHCO.sub.3 (400 mL) then extracted with EtOAc (2×300 mL). The combined organic layers were dried and concentrated under reduced pressure. The product was precipitated in DCM, filtered and washed with a minimal amount of DCM to give the desired product (4.75 g). The filtrate was purified by column chromatography on silica gel, using 0% to 50% EtOAc in cyclohexane as the eluent to give 1.4 g of the desired product. 2-(4-Aminophenyl)-1,1,1,3,3-pentafluoro-propan-2-ol (6.15 g, 68%) was obtained as an off-white solid.

(18) MS(ES.sup.+) m/z 242.0 [M+H].sup.+.

(19) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 5.27 (s, 2 H), 6.22-6.74 (m, 3 H), 7.11-7.35 (m, 3 H).

(20) Step 3:

(21) ##STR00014##

(22) To a solution of 2-(4-aminophenyl)-1,1,1,3,3,3-hexafluoro-propan-2-ol (0.55 g) in DCM (10 mL) were added 2-[4-(cyclopropylmethylsulfonyl)phenyl]acetic acid (0.58 g) and N-ethyl-N-isopropyl-propan-2-amine (1.19 mL) followed by dropwise addition of a 50% solution of T3P in DCM (0.8 mL). The reaction mixture was stirred at RT for 2 hours. EtOAc (100 mL) and water (100 mL) were added. The organic layer was separated, washed with brine (100 mL), dried over MgSO.sub.4, filtered and concentrated under reduced pressure. The residue was triturated in DCM (5 mL), filtered and washed with 10% DCM in pentane (5 mL) then dried to give 0.72 g (66%) of 2-[4-(cyclopropylmethylsulfonyl)phenyl]-N-[4-[1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide as an off-white solid.

(23) MS(ES.sup.+) m/z 478.1 [M+H].sup.+.

Examples 2 and 3

(24) The two enantiomers were separated from Example 1 by chiral chromatography using a column Chiralpak AD 20 μm, 76.5×350 mm and a mobile phase, EtOH:MeOH 90:10, 350 mL/min, with UV detection at 254 nm.

(25) Starting from 5 g of racemate in 500 mL of EtOH, five injections of the solution were done to yield, after concentration, 2.17 g of Example 2 (first enantiomer to be eluted) and 2.02 g of Example 3.

Example 2—Compound of (Formula IA): (+)-2-[4-(cyclopropylmethylsulfonyl)phenyl]-N-[4-[(1R)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide

(26) ##STR00015##

(27) MS(ES.sup.+) m/z 478.1 [M+H].sup.+.

(28) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 0.11 (m, 2 H), 0.44 (m, 2 H), 0.82 (m, 1 H), 3.24 (d, J=7.2 Hz, 2 H), 3.82 (s, 3 H), 6.67 (t, J=53.4 Hz, 2 H), 7.57 (d, J=9.2 Hz, 2 H), 7.60 (d, J=8.5 Hz, 2 H), 7.67 (d, J=9.2 Hz, 2 H), 7.70 (s, 1 H), 7.86 (d, J=8.5 Hz, 2 H), 10.42 (s, 1 H).

(29) Optical rotation: [α].sub.D.sup.20=+1.5° (c=3 mg/mL, DMSO).

Example 3—Compound of (Formula IB): (−)-2-[4-(cyclopropylmethylsulfonyl)phenyl]-N-[4-[(1S)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide

(30) ##STR00016##

(31) MS(ES.sup.+) m/z 478.0 [M+H].sup.+.

(32) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 0.11 (m, 2 H), 0.44 (m, 2 H), 0.82 (m, 1 H), 3.24 (d, J=7.2 Hz, 2 H), 3.82 (s, 3 H), 6.67 (t, J=53.4 Hz, 2 H), 7.56 (d, J=9.2 Hz, 2 H), 7.59 (d, J=8.5 Hz, 2 H), 7.67 (d, J=9.2 Hz, 2 H), 7.70 (s, 1 H), 7.86 (d, J=8.5 Hz, 2 H), 10.42 (s, 1 H).

(33) Optical rotation: [α].sub.D.sup.20=−4.6° (c=3 mg/mL, DMSO).

Example 4—X-Ray Single Crystal Diffraction

(34) The absolute configuration of Examples 2 and 3 has been determined from X-ray Single Crystal Diffraction (XRSCD) data using anomalous dispersion and Bijvoet differences analysis. A single crystal of suitable crystallographic quality for crystal structure determination from single crystal diffraction data has been isolated from a slow evaporation experiment of a solution of Examples 2 and 3 in chloroform.

(35) Absolute Configuration of Example 2

(36) Data have been collected on a Bruker Smart Apex single crystal diffractometer. A molybdenum IμS microfocus X-ray source has been used, running at 50 kV and 0.6 mA, emitting Mo-Kα radiation (λ=0.710731 Å). A Charge-Coupled Device (CCD chip: 4K, 62 mm) area detector has been positioned at 6.0 cm. An Oxford Cryosystems nitrogen cryostat (Cryostream Plus 700 series) has allowed XRSCD experiment to be carried out at 100 K. The single crystal with a size of: 25×250×250 μm.sup.3 has been mounted from a Paratone N™ oil drop onto a low background mylar MiTeGen loop. A full Ewald sphere of reflections has been collected (3 omega scans of 680 frames with a frame width of 0.3°). Accumulation time has been set at 80 seconds for each frame to be acquired.

(37) The orientation matrix and unit cell have been established using CELL_NOW (v2008/4) program. The 3D reflection profile and the integration of all reflections have been carried out with the SAINT (v8.34A) program. The TWINABS (v2012/1) program has been used to correct for Lorentz and polarization effects and for absorption by the sample. In addition, both HKLF4 and HKLF5 data has been generated respectively for solving and refining by the SHELXTL (v2014/7) program suite.

(38) The crystal structure data are as follows: Triclinic, space group P1 a=7.9892(13) Å, b=11.4301(18) Å, c=11.6936(19) Å a=83.022(2)°, β=77.006(2)°, γ=81.071(2)° V=1023.7(3) Å.sup.3, Z=2, T=100(2) K, 11704 reflections measured (3.6≤2⊖≤58.2), 11704 unique (R.sub.sigma=0.0229) 736 parameters and 3 restraints

(39) The final R.sub.1 is 2.7% (I>2σ(I)) and wR.sub.2 is 7.1% (all data) with Gof=1.030.

(40) The standard procedure for the determination of the absolute structure with X-ray diffraction techniques is based on the determination of the Flack parameter (x) with its associated standard uncertainty as part of the least-squares refinement procedure. Expected values are 0 (within 3 esd's) for correct and +1 for inverted absolute structure. In case of Example 2—compound of (Formula IA), Flack x=0.095(33) by classical fit to all intensities and 0.072(21) from 3969 selected quotients (Parsons' method giving a higher precision) indicate that Example 2—compound of (Formula IA) absolute configuration has been reliably determined.

(41) In addition a post-refinement procedure based on a Bayesian statistics approach was applied (a completely different way from Flack approach). Using a combination of maximum likelihood estimation and Bayesian statistics it not only gets a qualitative assignment of the absolute structure, but also a quantitative estimate of the reliability of that assignment. Analysis of Example 2—compound of (Formula IA) absolute structure using likelihood method has been performed using Olex2 software package. The resulting value is Hooft y=0.13(5) indicating that the absolute structure has been determined correctly (Bijvœt pair analysis using Student's t distribution with 88% Bijvœt pairs coverage: 4639 pairs used). The method has also calculated that the probability that the structure is inverted is equal to zero.

(42) Thus, these results indicate that the absolute configuration of Example 2—compound of (Formula IA) is R (probability of 100%).

(43) Absolute Configuration of Example 3

(44) Data have been collected on a Bruker Smart Apex single crystal diffractometer. A molybdenum IμS microfocus X-ray source has been used, running at 50 kV and 0.6 mA, emitting Mo-Kα radiation (λ=0.710731 Å). A Charge-Coupled Device (CCD chip: 4K, 62 mm) area detector has been positioned at 6.0 cm. An Oxford Cryosystems nitrogen cryostat (Cryostream Plus 700 series) has allowed XRSCD experiment to be carried out at 100 K. The single crystal with a size of: 40×150×200 μm.sup.3 has been mounted from a Paratone N™ oil drop onto a low background mylar MiTeGen loop. A full Ewald sphere of reflections has been collected (3 omega scans of 680 frames with a frame width of 0.3°). Accumulation time has been set at 75 seconds for each frame to be acquired.

(45) The orientation matrix and unit cell has been established using the Bruker AXS Apex2 (v2014.11 0) program suite. The 3D reflection profile and the integration of all reflections have been carried out with the SAINT (v8.34A) program. The SADABS (v2014/5) program has been used to correct for Lorentz and polarization effects and for absorption by the sample. The tentative space group has been determined with the XPREP (v2014/2) program. The SHELXTL XT (v2014/4) program has been used to solve the structure by the intrinsic phasing method. The SHELXTL XLMP (v2014/7) program has been used to refine the solution by full-matrix least-squares calculations on F.sup.2.

(46) The crystal structure data are as follows: Triclinic, space group P1 a=8.0022(5) Å, b=11.4394(8) Å, c=11.7044(8) Å α=83.0030(10)°, β=76.9840(10)°, γ=80.9850(10)° V=1026.83(12) Å.sup.3, Z=2, T=100(2) K 10430 reflections measured (3.6≤2⊖≤57.8), 8932 unique (R.sub.int=0.0081) 737 parameters and 3 restraints

(47) The final R.sub.1 was 2.5% (I>2σ(I)) and wR.sub.2 was 6.8% (all data) with Gof=1.024.

(48) The standard procedure for the determination of the absolute structure with X-ray diffraction techniques is based on the determination of the Flack parameter (x) with its associated standard uncertainty as part of the least-squares refinement procedure. Expected values are 0 (within 3 esd's) for correct and +1 for inverted absolute structure. In case of Example 3—compound of (Formula IB), Flack x=0.063(47) by classical fit to all intensities and 0.058(9) from 3969 selected quotients (Parsons' method giving a higher precision) indicate that Example 3—compound of (Formula IB) absolute configuration has been reliably determined.

(49) In addition a post-refinement procedure based on a Bayesian statistics approach was applied. Using a combination of maximum likelihood estimation and Bayesian statistics it not only gets a qualitative assignment of the absolute structure, but also a quantitative estimate of the reliability of that assignment.

(50) Analysis of Example 3—compound of (Formula IB) absolute structure using likelihood method has been performed using Olex2 software package. The resulting value is Hooft y=0.059(8) indicating that the absolute structure has been determined correctly (Bijvœt pair analysis using Student's t distribution with 75% Bijvœt pairs coverage: 4049 pairs used). The method has also calculated that the probability that the structure is inverted is equal to zero.

(51) Thus these results indicate that the absolute configuration of Example 3—compound of (Formula IB) is S (probability of 100%).

Example 5—RORγ GAL4 Reporter Gene Assay

(52) Examples 2-3 of the present compound of Formula IA and Compound of Formula IB application and example No. 37 from WO2015/082533 were tested for their ability to inhibit RORγ activity in a RORγ GAL4 reporter gene assay.

(53) The assay procedure is described below and results are presented in Table 1.

(54) A GAL4 one-hybrid reporter system employing luciferase readout was established to determine inhibition of RORγ in 293FT cells. The RORγ ligand-binding domain (LBD) was fused to the yeast GAL4 DNA binding domain (DBD) and placed under the control of the human cytomegalovirus (CMV) immediate early promoter, using expression vector pFN26A (Promega) and standard recombinant DNA cloning methods. To serve as a control in the assay, a similar vector was generated in which the GAL4-DBD was fused to Herpes simplex virus protein 16 (VP16), a constitutive transcriptional activator.

(55) To monitor the inhibitory effect of compounds on RORγ, a transcriptional reporter construct was used. The pGL4.35 vector (Promega) contains nine copies of the GAL4 Upstream Activator Sequence (UAS). This sequence drives the transcription of the luciferase reporter gene luc2P in response to binding of a fusion protein containing the GAL4 DNA binding domain, as for example expressed by the GAL4-RORγ-LBD and GAL4-VP16 expression vectors described above. To allow a GAL4 fusion protein to drive the expression of the luciferase reporter, the pGL4.35 expression vector and the appropriate GAL4 fusion protein expression vector were bulk transfected in the 293FT cells using standard transfection techniques.

(56) The day after transfection, cells were plated into 96 well plates, test compound was added and the plates were incubated overnight. Subsequently, the firefly luciferase activity was quantified using luciferase detection reagent and luminescence readout.

(57) Detailed Assay Description

(58) 293FT cells (Invitrogen) were transfected with a GAL4 fusion protein expression vector (as described above) and the transcriptional reporter construct (pGL4.35, Promega). 60 μL of TransIT-293 transfection reagent (Mirus Bio) was added drop wise to 1500 μl Opti-MEM I Reduced Serum Medium (Invitrogen) and incubated at room temperature (RT) for 5 to 20 minutes. 1500 μL of this reagent mixture was added to 5 μg of GAL4 fusion protein expression vector and 5 μg of the transcriptional reporter construct, and incubated at RT for 20 minutes.

(59) To harvest 293FT cells from a T75 flask, first the culture medium was taken off the cells. Subsequently, the cells were washed with Phosphate Buffered Saline (PBS) (Lonza), after which the PBS was removed. To dissociate the cells, 1 ml of TrypLE Express (Invitrogen) was added to the flask, followed by incubation at RT until the cells visually started to detach. Cells were collected in 5 mL of assay medium (DMEM culture medium (Lonza), 10% dialyzed FBS (Invitrogen) and Pen/Strep (Lonza)) to achieve a single cell suspension. 10×10.sup.6 cells were spun down and re-suspended in 10 mL of assay medium. Subsequently, the cell suspension was added to the transfection mix tube, and then transferred as a whole to a T75 flask (Greiner), followed by overnight (16-24 hours) incubation at 37° C. and 5% CO.sub.2.

(60) For compound screening, the cells were harvested (as described above) and counted. 13×10.sup.6 cells were spun down, the supernatant was aspirated and the cells were re-suspended in 17.3 mL of assay medium to obtain a cell suspension of 0.75×10.sup.6 cells/mL. 80 μL of cell suspension (60,000 cells) was plated per well into a white, flat bottom, tissue culture treated, 96 well screening plates (Greiner).

(61) Test compounds were diluted, starting from a 10 mM dimethylsulfoxide (DMSO) stock solution, to serial dilutions in DMSO at 500× the final test concentration. Subsequently, these solutions were diluted to 5× the final test concentration in two 10-fold-dilution steps in assay medium. The final DMSO concentration of the 5× test compound solution was 1%. 20 μL of the 5× test compound solution was added to each test well of the 96 well plate previously plated with 80 μl cell suspension, resulting in the final test concentration with 0.2% DMSO.

(62) The plates were incubated overnight (16-24 hours) at 37° C. and 5% CO.sub.2.

(63) For the luciferase readout, the luciferase reagent (Britelite Plus, Perkin Elmer) was brought to RT. To each test well of the screening plates, 100 μL of 2.5-fold diluted Britelite Plus reagent was added, followed by incubation at RT for 10 minutes. The luciferase luminescence signal was measured using a Wallac Victor Microplate Reader (Perkin Elmer).

(64) The half maximum inhibitory concentration (IC.sub.50) values for the test compounds were calculated from the luciferase signal using GraphPad Prism software (GraphPad Software).

Example 6—Peripheral Blood Mononuclear Cell (PBMC) IL-17 Assay

(65) Examples 2-3 of the present application and example No. 37 from WO2015/082533 were tested for their ability to inhibit the IL-17A production in anti-CD3/anti-CD28 stimulated peripheral blood mononuclear cells (PBMCs) isolated from human blood. The assay procedure is described below and results are presented in Table 1.

(66) This assay is designed to measure the levels of IL-17A secreted from anti-CD3/anti-CD28 stimulated PBMCs with the aim of measuring the inhibition of RORγ mediated IL-17A production.

(67) Detailed Assay Description

(68) The assay medium consists of 90% RPMI 1640 (Lonza), 10% heat-inactivated fetal bovin serum (FBS, Lonza) and 100 U/mL penicillin/streptomycin solution.

(69) Anti-CD3 antibody (BD Pharmingen) was diluted to 10 μg/ml in PBS (Lonza). 30 μL of 10 μg/ml anti-CD3 solution was added to the inner 60 wells of a 96-well cell culture treated U-bottom plate (Greiner). Plates were incubated overnight (16-24 hours) at 37° C. and 5% CO.sub.2.

(70) Peripheral blood mononuclear cells were separated from buffy coats (Sanquin) using Ficoll-Paque PREMIUM separation medium (GE Healthcare Life Sciences) according to manufacturer's protocol and re-suspended in assay medium at 37° C.

(71) Test compounds were diluted, starting from a 10 mM dimethylsulfoxide (DMSO) stock solution, to serial dilutions in DMSO at 200× the final test concentration. Subsequently, these solutions were diluted in two dilution steps in assay medium to 5× the final test concentration. The DMSO concentration of the 5× test compound solution was 2.5%.

(72) Anti-CD28 antibody (BD Pharmingen) was diluted to 10 μg/mL in assay medium. The PBMCs were diluted to a concentration of 2.2×10.sup.6 cells/mL in assay medium at 37° C.

(73) For compound screening, the anti-CD3 coated plates were washed two times with PBS; the wells were subsequently aspirated using vacuum. To each screening well 90 μL of the PBMC suspension, 30 μL of the anti-CD28 solution and 30 μL of the 5× test compound solution was added, resulting in the final test concentration with 0.5% DMSO. All outer wells were filled with PBS to prevent evaporation. Plates were incubated for 5 days at 37° C. and 5% CO.sub.2.

(74) After incubation, the plates were spun down at 1500 rpm for 4 minutes and the supernatant was collected. Subsequently, the IL-17A levels in the supernatants was determined using an IL-17A AlphaLISA kit (Perkin Elmer) according to the manufacturer's protocol.

(75) The half maximum inhibitory concentration (IC.sub.50) values for the test compounds were calculated from the IL-17A signal using GraphPad Prism software (GraphPad Software).

(76) TABLE-US-00001 TABLE 1 RORγ GAL4 reporter gene assay pIC.sub.50 PBMC IL-17 assay pIC.sub.50 Standard Number of Standard Number of Example No Average deviation experiments Average deviation human donors 37 7.9 0.2 5 7.9 0.2 11 2 7.4 0.2 5 7.2 0.2 11 3 7.9 0.2 5 7.6 0.3 11

Example 7—hERG Channel Protocol

(77) Examples 2-3 of the present application and example No. 37 of WO2015/082533 were tested in vitro against the hERG (human Ether-à-go-go-Related Gene) potassium channel. The assay procedure is described below and results are presented in Table 2.

(78) Detailed Assay Description

(79) Frozen CHO (Chinese hamster ovary) cells stably expressing hERG channels were thawed and seeded on glass coverslips in petri dishes. Cells were cultured in HAM's F-12 media supplemented with 10% fetal bovine serum, 100 IU/mL penicillin, 100 μg/mL streptomycin and 500 μg/mL G418 (Invitrogen, Carlsbad, Calif.) in an atmosphere of 95% air/5% CO.sub.2 at 37° C. CHO cells were ready for patch-clamping after culture for 1-5 days. hERG currents were recorded at room temperature using the whole-cell patch-clamp technique with an Axopatch 200B amplifier (Molecular Devices, Sunnyvale Calif.). Electrodes (1-3 MΩ resistance) were fashioned from TW150F glass capillary tubes (World Precision Instruments, Sarasota, Fla.) and filled with a solution containing (in mM): potassium aspartate 120; KCl 20; Na.sub.2ATP 4; HEPES 5; MgCl.sub.2 1; pH 7.2 adjusted with KOH. The external recording solution contained (in mM): NaCl 130; KCl 4; sodium acetate 2.8; MgCl.sub.2 1; HEPES, 10; glucose 10; CaCl.sub.2 1 at pH 7.4 adjusted with NaOH. hERG currents were elicited by 2 s depolarizing pulses to +20 mV followed by repolarization to −40 mV for 1.6 s from a −80 mV holding potential at a frequency of 0.1 Hz. Currents were analyzed using the pCLAMP suite of software (Molecular Devices). IC.sub.50 values of drugs were obtained using peak tail currents during the −40 mV step by nonlinear least-squares fit of the data (GraphPad Software, Inc. San Diego, Calif.).

Example 8—CYP3A4 Inhibition Protocol

(80) Examples 2-3 of the present application and example No. 37 from WO2015/082533 were tested for CYP3A4 inhibition using two different probes: midazolam and testosterone.

(81) The assay procedure is described below and results are presented in Table 2.

(82) Detailed Assay Description

(83) Test compound dissolved in DMSO at selected concentrations was added to a phosphate buffer (50 mM, pH 7.4) containing human liver microsomes (0.1 mg/mL), MgCl (6 mM) and EDTA (0.5 mM) and CYP3A probe substrate, either midazolam (3 μM). Test compound was evaluated at the concentrations 1, 3, 10 and 30 μM and the final DMSO concentration in the incubation was 0.5%. After addition of 1 M NADPH the mixture was incubated at 37° C. for 10 min (midazolam) or 30 min (testosterone). The reaction was terminated by addition of cold CH3CN containing internal standard, centrifuged and formation of the CYP3A specific metabolite (either 1′-hydroxymidazolam or 6-β-hydroxytestosterone) was quantified by UPLC-MS/MS. The relative CYP3A activity at each test concentration was calculated and IC.sub.50 values determined using XLfit.

(84) TABLE-US-00002 TABLE 2 CYP3A4 inhibition Assay IC.sub.50, μM Midazolam Testosterone hERG inhibition Example No probe probe Assay IC.sub.50, μM 37 10 2.8 4.3 2 24 13 14.2 3 >30 16 11.2

(85) The present application provides novel RORγ modulator compounds, Examples 2 and 3, which inhibit RORγ activity and for which both hERG inhibition and CYP3A4 inhibition were decreased when compared to prior art compound (example 37 from WO2015/082533). It therefore appears that compounds of (Formula IA) and of (Formula IB) limit the risks of cardiac safety issues and potential adverse toxic effects related to drug-drug interactions respectively while maintaining the RORγ modulation activity.

Example 9, 10, 11, 12 and 13: Synthesis of the Prodrugs

Example 9—Compound of (Formula VI): 2-[4-(cyclopropylmethylsulfinyl)phenyl]-N-[4-[1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide

(86) ##STR00017##

(87) Step 1:

(88) ##STR00018##

(89) In a 100 mL three-neck round bottom flask placed under argon, hydrogen peroxide (2.64 mL, 25.88 mmol, 30% in water) was added dropwise to a solution of ethyl 2-[4-(cyclopropylmethylsulfanyl)phenyl]acetate (3 g, 11.98 mmol) in 1,1,1,3,3,3-hexafluoropropan-2-ol (12.53 mL, 113.84 mmol). The reaction mixture was stirred for 30 minutes at RT, then a 10% aqueous solution of sodium thiosulfate (50 mL) was added, followed by brine (10 mL). The aqueous layer was extracted twice with EtOAc. The combined organic extracts were washed with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under reduced pressure. The residue was taken into ACN (15 mL), concentrated under reduced pressure and dried under high vacuum to obtain 3.16 g of ethyl 2-[4-(cyclopropylmethylsulfinyl)phenyl]acetate as a white solid.

(90) MS(ES.sup.+) m/z 267.1 [M+H].sup.+.

(91) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 7.62 (d, J=8.28 Hz, 2 H), 7.46 (d, J=8.28 Hz, 2 H), 4.09 (q, J=7.03 Hz, 2 H), 3.76 (s, 2 H), 2.72-2.87 (m, 2 H), 1.18 (t, J=7.15 Hz, 3 H), 0.82-0.96 (m, 1 H), 0.45-0.61 (m, 2 H), 0.19-0.36 (m, 2 H).

(92) Step 2:

(93) ##STR00019##

(94) In a 100 mL round bottom flask, a 1M aqueous solution of sodium hydroxide (5.41 mL, 5.41 mmol) was added to a solution of ethyl 2-[4-(cyclopropylmethylsulfinyl)phenyl]acetate (400 mg, 1.50 mmol) in EtOH (25 mL). The reaction mixture was stirred overnight at RT and then concentrated under vacuum. The residue was taken into water, then acidified with an aqueous solution of HCl 1N until reaching pH 1. The aqueous layer was extracted three times with EtOAc. The combined organic layers were washed once with brine, dried over Na.sub.2SO.sub.4, filtered and concentrated under vacuum to obtain 306 mg of 2-[4-(cyclopropylmethylsulfinyl)phenyl]acetic acid as a white solid.

(95) MS(ES+) m/z 239.1 [M+H]+.

(96) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 7.61 (d, J=8.28 Hz, 2 H), 7.46 (d, J=8.28 Hz, 2 H), 3.66 (s, 2 H), 2.71-2.89 (m, 2 H), 0.85-0.96 (m, 1 H), 0.46-0.64 (m, 2 H), 0.20-0.36 (m, 2 H).

(97) Step 3:

(98) ##STR00020##

(99) 2-(4-aminophenyl)-1,1,1,3,3-pentafluoro-propan-2-ol (151.8 mg, 0.63 mmol), N-ethyl-N-isopropyl-propan-2-amine (0.33 mL, 1.89 mmol) and 2,4,6-Tripropyl-1,3,5,2,4,6-trioxatriphosphinane 2,4,6-trioxide (50% in DCM, 0.48 mL, 0.82 mmol, dropwise) were added to a solution of 2-[4-(cyclopropylmethylsulfinyl)phenyl]acetic acid (150 mg, 0.63 mmol) in DCM (25 mL), which had been prealably placed under argon in an ice bath. After stirring for 30 minutes, the ice bath was removed and the reaction mixture was stirred overnight at RT. Then, DCM and water were added, and the aqueous layer was extracted twice with DCM. The combined organic layers were washed with a saturated aqueous solution of NaHCO.sub.3, with a 0.5N aqueous HCl solution, with water, and finally with brine. The organic layer was dried with Na.sub.2SO.sub.4, filtered and concentrated under vacuum. The residue was purified by column chromatography on silica gel, using from 98% to 95% DCM in MeOH as the eluent to give 206 mg of 2-[4-(cyclopropylmethylsulfinyl)phenyl]-N-[4-[1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide as an off-white foam.

(100) MS(ES.sup.+) m/z 462.0 [M+H].sup.+.

(101) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 10.35 (s, 1 H), 7.45-7.79 (m, 9 H), 6.38-6.88 (m, 1 H), 3.75 (s, 2 H), 2.67-2.90 (m, 2 H), 0.75-0.96 (m, 1 H), 0.45-0.62 (m, 2 H), 0.14-0.37 (m, 2 H).

Examples 10, 11, 12 and 13

(102) The stereoisomers VIA and VID were separated from VI by chiral chromatography using a column Chiralpak AD 20 μm, 350×76.5 mm and a mobile phase, Heptane:EtOH 50:50, 400 mL/min with UV detection at 254 nm.

(103) Starting from 160 mg of racemate in 100 mL of Heptane:EtOH 50:50, one injection of the solution was done to yield, after concentration, 34 mg of Example 10 (first enantiomer to be eluted), 35 mg of Example 13 (last enantiomer to be eluted), and 70 mg of a mixture of Examples 11 and 12.

(104) The stereoisomers VIB and VIC were separated from the corresponding mixture by chiral chromatography using a column Cellulose-4 5 μm, 250×4.6 mm and a mobile phase, Heptane:EtOH 70:30, 45 mL/min with UV detection at 254 nm.

(105) Starting from 70 mg of the mixture in 8 mL of EtOH, four injections of the solution was done to yield, after concentration, 29 mg of VIB (first enantiomer to be eluted), and 30 mg of VIC.

Example 10—Compound of (Formula VIA): (−)-2-[4-[(S)-cyclopropylmethylsulfinyl]phenyl]-N-[4-[(1R)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide

(106) Absolute configuration was assigned arbitrarily.

(107) ##STR00021##

(108) MS(ES.sup.+) m/z 462.2 [M+H].sup.+.

(109) .sup.1H NMR (400 MHz, DMSO-d6) δ ppm 10.35 (s, 1 H), 7.66 (m, 3 H), 7.63 (d, J=8.4 Hz, 2 H), 7.56 (d, J=9.0 Hz, 2 H), 7.52 (d, J=8.4 Hz, 2 H), 6.64 (t, J=53.2 Hz, 1 H), 3.75 (s, 2 H), 2.82 (dd, J=6.9 et 13.3 Hz, 1 H), 2.74 (dd, J=7.7 et 13.3 Hz, 1 H), 0.89 (m, 1 H), 0.49 a 0.58 (m, 2 H), 0.23 à 0.33 (m, 2 H).

(110) Optical rotation: [α].sub.D.sup.20=−63.6° (c=3.8 mg/mL, DMSO).

Example 11—Compound of (Formula VIB): (−)-2-[4-[(S)-cyclopropylmethylsulfinyl]phenyl]-N-[4-[(1S)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide

(111) Absolute configuration was assigned arbitrarily.

(112) ##STR00022##

(113) MS(ES.sup.+) m/z 462.2 [M+H].sup.+.

(114) .sup.1H NMR (400 MHz, DMSO-d6): δ 10.35 (s, 1 H), 7.66 (m, 3 H), 7.63 (d, J=8.4 Hz, 2 H), 7.56 (d, J=9.0 Hz, 2 H), 7.52 (d, J=8.4 Hz, 2 H), 6.64 (t, J=53.2 Hz, 1 H), 3.75 (s, 2 H), 2.82 (dd, J=6.9 et 13.3 Hz, 1 H), 2.74 (dd, J=7.7 et 13.3 Hz, 1 H), 0.89 (m, 1 H), 0.49 a 0.58 (m, 2 H), 0.23 à 0.33 (m, 2 H).

(115) Optical rotation: [α].sub.D.sup.20=−83.1° (c=4.2 mg/mL, DMSO).

Example 12—Compound of (Formula VIC): (+)-2-[4-[(R)-cyclopropylmethylsulfinyl]phenyl]-N-[4-[(1R)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide

(116) Absolute configuration was assigned arbitrarily.

(117) ##STR00023##

(118) MS(ES.sup.+) m/z 462.0 [M+H].sup.+.

(119) .sup.1H NMR (400 MHz, DMSO-d6): δ 10.35 (s, 1 H), 7.66 (m, 3 H), 7.63 (d, J=8.4 Hz, 2 H), 7.56 (d, J=9.0 Hz, 2 H), 7.52 (d, J=8.4 Hz, 2 H), 6.64 (t, J=53.4 Hz, 1 H), 3.75 (s, 2 H), 2.82 (dd, J=6.9 et 13.3 Hz, 1 H), 2.74 (dd, J=7.7 et 13.3 Hz, 1 H), 0.89 (m, 1 H), 0.49 a 0.58 (m, 2 H), 0.23 a 0.33 (m, 2 H).

(120) Optical rotation: [α].sub.D.sup.20=+85.6° (c=5.8 mg/mL, DMSO).

Example 13—Compound of (Formula VID): (+)-2-[4-[(R)-cyclopropylmethylsulfinyl]phenyl]-N-[4-[(1S)-1-(difluoromethyl)-2,2,2-trifluoro-1-hydroxy-ethyl]phenyl]acetamide

(121) Absolute configuration was assigned arbitrarily.

(122) ##STR00024##

(123) MS(ES.sup.+) m/z 462.0 [M+H].sup.+.

(124) .sup.1H NMR (400 MHz, DMSO-d6): δ 10.35 (s, 1 H), 7.66 (m, 3 H), 7.63 (d, J=8.4 Hz, 2 H), 7.56 (d, J=9.0 Hz, 2 H), 7.52 (d, J=8.4 Hz, 2 H), 6.64 (t, J=53.4 Hz, 1 H), 3.75 (s, 2 H), 2.82 (dd, J=6.9 et 13.3 Hz, 1 H), 2.74 (dd, J=7.7 et 13.3 Hz, 1 H), 0.89 (m, 1 H), 0.49 à 0.58 (m, 2 H), 0.23 à 0.33 (m, 2 H).

(125) Optical rotation: [α].sub.D.sup.20=+60.9° (c=3.8 mg/mL, DMSO).

Example 14—Aqueous Equilibrated Solubility

(126) Sample Preparation

(127) The studied compound was accurately weighted with a target concentration of 2 mg/mL in an aqueous phosphate buffer (50 mM at pH=7.4). The solution was shaken overnight (rock'n roll shaker) at RT and protected from light (about 24 hrs). The solution was filtered in plate filter device (microplate Millipore “Solvinert” with integrated PTFE filter; 0.45 μm) and the filtrate was dosed by LC/UV method.

(128) Reference (Standard) Preparation

(129) The studied compound was accurately weighted with a target concentration of 0.1 mg/mL in DMSO. The solution was sonicated at RT and protected from light. The reference solution was dosed by a LC/UV method and pH of the solubilized fraction was measured.

(130) TABLE-US-00003 TABLE 3 Example No Solubility pH 7.5* (μg/mL) 2 <1.0 3 <1.0 9 99.4 10 94.8 11 92.5 12 94.4 13 93.0 *pH of the solubilized fraction.