WO2012028100A1 - Novel compounds - Google Patents

Abstract

Novel retinoid-related orphan receptor gamma (ROR_ϒ) modulators and their use in the treatment of diseases mediated by ROR_ϒ provided by the present invention.

Description

NOVEL COMPOUNDS

The present invention relates to novel retinoid-related orphan receptor gamma (RORy) modulators and their use in the treatment of diseases mediated by RORy.

Background of the Invention Retinoid-related orphan receptors (RORs) are transcription factors which belong to the steroid hormone nuclear receptor superfamily (Jetten & Joo (2006) Adv. Dev. Biol. 16:313-355). The ROR family consists of three members, ROR alpha (RORa), ROR beta (RORP) and ROR gamma (RORy), each encoded by a separate gene (RORA, RORB and RORC, respectively). RORs contain four principal domains shared by the majority of nuclear receptors: an N-terminal A B domain, a DNA-binding domain, a hinge domain, and a ligand binding domain. Each ROR gene generates several isoforms which differ only in their N-terminal A/B domain. Two isoforms of RORy have been identified: RORyl and RORyt (also known as RORy2). RORy is a term used to describe both RORyl and/or RORyt.

While RORyl is expressed in a variety of tissues including thymus, muscle, kidney and liver, RORyt is exclusively expressed in the cells of the immune system. RORyt has been identified as a key regulator of Thl7 cell differentiation. Thl7 cells are a subset of T helper cells which produce IL- 17 and other proinflammatory cytokines. Thl7 cells have been shown to have key functions in several mouse autoimmune disease models including experimental autoimmune encephalomyelitis (EAE) and collagen-induced arthritis (CIA), In addition, Thl7 cells or their products have been shown to be associated with the pathology of a variety of human inflammatory and autoimmune disorders including multiple sclerosis, rheumatoid arthritis, psoriasis, Crohn's disease and asthma (Jetten (2009) Nucl. Recept. Signal. 7: e003; Manel et al. (2008) Nat. Immunol. 9:641 -649). The pathogenesis of chronic autoimmune diseases including multiple sclerosis and rheumatoid arthritis arises from the break in tolerance towards self-antigens and the development of auto-aggressive effector T cells infiltrating the target tissues. Studies have shown that Thl7 cells are one of the important drivers of the inflammatory process in tissue-specific autoimmunity (Steinman (2008) J. Exp. Med. 205: 1517-1522; Leung et al. (2010) Cell. Mol. Immunol. 7: 182-189). There is evidence that Thl7 cells are activated during the disease process and are responsible for recruiting other inflammatory cells types, especially neutrophils, to mediate pathology in the target tissues (Korn et al. (2009) Annu. Rev. Immunol. 27:485-517). RORyt plays a critical role in the pathogenic responses of Thl7 cells (Ivanov et al. (2006) Celt 126: 1 121-1133). RORyt deficient mice show very little TM7 cells. In addition, RORyt deficiency resulted in amelioration of EAE. Further support for the role of RORyt in the pathogensis of autoimmune or inflammatory diseases can be found in the following references: Jetten & Joo (2006) Adv.Dev.Biol. 16:313-355; Meier et al. (2007) Immunity 26:643-654; Aloisi & Pujol-Borrell (2006) Nat. Rev. Immunol, 6:205-217; Jager et al. (2009) J Immunol. 183:7169-7177; Serafmi et al. (2004) Brain Pathol.14: 164-174; Magliozzi et al. (2007) Brain 130: 1089-1 104; Barnes (2008) Nat.Rev.Immunol. 8: 183-192.

In light of the role RORy plays in the pathogenesis of diseases, it is desirable to prepare compounds that modulate RORy activity, which can be used in the treatment of diseases mediated by RORy.

Summary of the Invention

The invention is directed to novel RORy modulators and their use in the treatment of diseases mediated by RORy. Specifically, the invention is directed to compounds according to Formula 1(a).

Formula 1(a)

wherein Rl , R2, and R3 are defined below, and to pharmaceutically-acceptable salts thereof.

In another aspect, this invention provides for the use of the compounds of Formula 1(a) for the treatment of diseases mediated by RORy. Examples of such diseases include autoimmune or inflammatory diseases such as multiple sclerosis, rheumatoid arthritis, psoriasis, Crohn's disease and asthma. In yet another aspect, the invention is directed to methods of treating such diseases.

In yet another aspect, this invention provides for the use of the compounds according to Formula I or the pharmaceutically-acceptable salts thereof for the treatment of diseases mediated by RORy.

Formula I

wherein Rl, R2, and R3 are defined below.

Brief Description of the Figures

Figure 1 (A) shows the inhibitory effect of the RORy modulator Example 2 on the production of IL- 17 by ELISA.

Figure 1 (B) shows the inhibitory effect of the RORy modulator Example 2 on the production of IL- 17 by intracellular staining.

Figure 2 shows the mean clinical scores of control and EAE mice treated with the RORy modulator Example 2.

Detailed Description of the Invention

Terms and Definitions

In describing the invention, chemical elements are identified in accordance with the Periodic Table of the Elements.

"Alkyl" refers to a monovalent saturated hydrocarbon chain having the specified number of member atoms. For example, C1-C6 alkyl refers to an alkyl group having from 1 to 6 member atoms. Alkyl groups may be optionally substituted with one or more substituent as defined herein. Alkyl groups may be straight or branched. Representative branched alkyl groups have one, two, or three branches. Alkyl includes methyl, ethyl, propyl (n-propyl and isopropyl), butyl (n-butyl, isobutyl, and t-butyl), pentyl (n-pentyl, isopentyl, and neopentyl), and hexyl.

"Alkoxy" refers to the group -O-R where R is alkyl having the specified number of member atoms. Alkoxy includes methoxy, ethoxy and propoxy.

"Enantiomerically enriched" refers to products whose enantiomeric excess is greater than zero. For example, enantiomerically enriched refers to products whose enantiomeric excess is greater than 50% ee, greater than 75% ee.and greater than 90% ee. "Enantiomeric excess" or "ee" is the excess of one enantiomer over the other expressed as a percentage. As a result, since both enantiomers are present in equal amounts in a racemic mixture, the enantiomeric excess is zero (0% ee). However, if one enantiomer was enriched such that it constitutes 95% of the product, then the enantiomeric excess would be 90% ee (the amount of the enriched enantiomer, 95%, minus the amount of the other enantiomer, 5%).

"Enantiomerically pure" refers to products whose enantiomeric excess is 99% ee or greater.

"Half-life" refers to the time required for half of a quantity of a substance to be converted to another chemically distinct species in vitro or in vivo.

"Halo" refers to the halogen radicals fluoro, chloro, bromo, and iodo.

"Heteroatom" refers to a nitrogen, sulphur, or oxygen atom.

"Heterocycloalkyl" refers to a saturated or unsaturated ring containing from 1 to 4 heteroatoms as member atoms in the ring. However, heterocycloalkyl rings are not aromatic.

Heterocycloalkyl groups containing more than one heteroatom may contain different heteroatoms. Heterocycloalkyl groups may be optionally substituted with one or more substituent as defined herein. Heterocycloalkyl groups are monocyclic ring systems or are fused, spiro, or bridged bicyclic ring systems. Monocyclic heterocycloalkyl rings have from 5 to 7 member atoms. Bicyclic

heterocycloalkyl rings have from 7 to 11 member atoms. In certain embodiments, heterocycloalkyl is saturated. In other embodiments, heterocycloalkyl is unsaturated but not aromatic. Heterocycloalkyl includes pyrrolidinyl, tetrahydrofuranyi, dihydrofuranyl, pyranyl, tetrahydropyranyl, dihydropyranyl, tetrahydrothienyl, pyrazolidinyl, oxazolidinyl, thiazolidinyl, piperidinyl, homopiperidinyl, piperazinyl, morpholinyl, thiamorpholinyl, azepinyl, 1,3-dioxolanyI, 1 ,3-dioxanyl, 1 ,4-dioxanyl, 1,3- oxathiolanyl, 1 ,3-oxathianyl, 1 ,3-dithianyl, azetidinyl, azabicylo[3.2.1]octyl, azabicylo[3.3.1]nonyl, azabicylo[4.3.0]nonyl, and oxabicylo[2.2.1]heptyl.

"Member atoms" refers to the atom or atoms that form a chain or ring. Where more than one member atom is present in a chain and within a ring, each member atom is covalently bound to an adj cent member atom in the chain or ring. Atoms that make up a substituent group on a chain or ring are not member atoms in the chain or ring.

"Optionally substituted" indicates that a group, such as alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycioalkenyl, heterocycloalkyl, or heteroaryl, may be unsubstituted, or the group may be substituted with one or more substituent as defined. "RORy" refers to all isoforms encoded by the RORC gene which include RORyl and

RORyt.

"RORy modulator" refers to a chemical compound that inhibits, either directly or indirectly, the activity of RORy. RORy modulators include antagonists and inverse agonists of RORy.

"Pharmaceutically acceptable" refers to those compounds, materials, compositions, and dosage fonns which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, or other problem or complication, commensurate with a reasonable benefit/risk ratio.

"Substituted" in reference to a group indicates that one or more hydrogen atom attached to a member atom within the group is replaced with a substituent selected from the group of defined substituents. It should be understood that the term "substituted" includes the implicit provision that such substitution be in accordance with the permitted valence of the substituted atom and the substituent and that the substitution results in a stable compound (i.e. one that does not

spontaneously undergo transformation such as by rearrangement, cyclization, or elimination and that is sufficiently robust to survive isolation from a reaction mixture). When it is stated that a group may contain one or more substituent, one or more (as appropriate) member atom within the group may be substituted. In addition, a single member atom within the group may be substituted with more than one substituent as long as such substitution is in accordance with the permitted valence of the atom. Suitable substituents are defined herein for each substituted or optionally substituted group.

Compounds

The present invention provides, in a first aspect, a compound of Formula 1(a) or a

pharmaceutically acceptable salt thereof.

Formula 1(a)

wherein:

Rl is H; R2 is H; and R3 is selected from the group consisting of:

- C1-C6 alkyl substituted with OH, C1-C3 alkoxy or one to three F; and

- NHRa wherein Ra is H or C1-C3 alkyl.

or

Rl is H or C1 -C3 alkyl, R2 is H or C1-C3 alkyl, wherein at least one of Rl and R2 is not H; and

R3 is Cl-C6 alkyl.

In one embodiment, the present invention relates to compounds of Formula 1(a) wherein Rl is H, R2 is H, and R3 is methylamino or ethylamino.

In another embodiment, the present invention relates to compounds of Formula 1(a) wherein

Rl is H, R2 is H, and R3 is C1-C3 alkyl substituted with OH, C1-C3 alkoxy or CF₃.

In another embodiment, the present invention relates to compounds of Formula 1(a) wherein at least one of Rl and R2 is not H, and R3 is C1-C3 alkyl.

In another embodiment, the present invention relates to compounds of Formula 1(a) wherein at least one of Rl and R2 is methyl, and R3 is C1-C3 alkyl.

In another aspect, the present invention provides for the use of the compounds according to Formula I for the treatment of diseases mediated by RORy.

Formula I

wherein:

Rl is H or CI-C6 alkyl;

R2 is H or Cl-C6 alkyl; and

R3 is selected from the group consisting of:

- C1-C6 alkyl optionally substituted with OH, C1-C3 alkoxy or one to three F; and

- NHRa wherein Ra is H or CI -C3 alkyl. The meaning of any functional group or substituent thereon at any one occurrence in Formula I, or any subformula thereof, is independent of its meaning, or any other functional group's or substituent's meaning, at any other occurrence, unless stated otherwise.

The compounds according to Formula I may contain one or more asymmetric center (also referred to as a chiral center) and may, therefore, exist as individual enantiomers, diastereomers, or other stereoisomeric forms, or as mixtures thereof. Chiral centers, such as chiral carbon atoms, may also be present in a substituent such as an alkyl group. Where the stereochemistry of a chiral center present in Formula I, or in any chemical structure illustrated herein, is not specified the structure is intended to encompass all individual stereoisomers and all mixtures thereof. Thus, compounds according to Formula I containing one or more chiral center may be used as racemic mixtures, enaniiomerically enriched mixtures, or as enantiomerically pure individual stereoisomers.

Individual stereoisomers of a compound according to Formula I which contain one or more asymmetric center may be resolved by methods known to those skilled in the art. For example, such resolution may be carried out (1) by formation of diastereoisomeric salts, complexes or other derivatives; (2) by selective reaction with a stereoisomer-specific reagent, for example by enzamatic oxidation or reduction; or (3) by gas-liquid or liquid chromatography in a chiral envtornment, for example, on a chiral support such as silica with a bound chiral ligand or in the presence of a chiral solvent. The skilled artisan will appreciate that where the desired stereoisomer is converted into another chemical entity by one of the separation procedures described above, a further step is required to liberate the desired form. Alternatively, specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to the other by asymmetric transformation.

The compounds according to Formula I may also contain double bonds or other centers of geometric asymmetry. Where the stereochemistry of a center of geometric asymmetry present in Formula I, or in any chemical structure illustrated herein, is not specified, the structure is intended to encompass the trans (E) geometric isomer, the cis (Z) geometric isomer, and all mixtures thereof. Likewise, all tautomeric forms are also included in Formula I whether such tautomers exist in equilibrium or predominately in one form.

In certain embodiments, compounds according to Formula I may contain an acidic functional group. In certain other embodiments, compounds according to Formula I may contain a basic functional group. Thus, the skilled artisan will appreciate that pharmaceutically-acceptable salts of the compounds according to Formula I may be prepared. Indeed, in certain embodiments of the invention, pharmaceutically-acceptable salts of the compounds according to Formula I may be preferred over the respective free base or free acid because such salts may impart greater stability or solubility to the molecule thereby facilitating formulation into a dosage form. Accordingly, the invention is further directed to the use of pharmaceutically-acceptable salts of the compounds according to Formula I.

As used herein, the term "pharmaceutically-acceptable salts" refers to salts that retain the desired biological activity of the subject compound and exhibit minimal undesired toxicological effects. These pharmaceutically-acceptable salts may be prepared in situ during the final isolation and purification of the compound, or by separately reacting the purified compound in its free acid or free base form with a suitable base or acid, respectively.

As used herein, the term "compounds of the invention" means both the compounds according to Formula I and the pharmaceutically-acceptable salts thereof. The term "a compound of the invention" also appears herein and refers to both a compound according to Formula I and its pharmaceutically-acceptable salts.

The invention also includes various deuterated forms of the compounds of Formula (I). Each available hydrogen atom attached to a carbon atom may be independently replaced with a deuterium atom. A person of ordinary skill in the art will know how to synthesize deuterated forms of the compounds of Formula (I). Commercially available deuterated starting materials may be employed in the preparation of deuterated forms of the compounds of Formula (I), or they may be synthesized using conventional techniques employing deuterated reagents (e.g. lithium aluminum deuteride).

The compounds of the invention may exist in solid or liquid form. In the solid state, the compounds of the invention may exist in crystalline or noncrystalline form, or as a mixture thereof. For compounds of the invention that are in crystalline form, the skilled artisan will appreciate that phannaceutically-acceptable solvates may be formed wherein solvent molecules are incorporated into the crystalline lattice during crystallization. Solvates may involve nonaqueous solvents such as ethanol, isopropanol, DMSO, acetic acid, ethanolamine, and ethyl acetate, or they may involve water as the solvent that is incorporated into the crystalline lattice. Solvates wherein water is the solvent that is incorporated into the crystalline lattice are typically referred to as "hydrates." Hydrates include stoichiometric hydrates as well as compositions containing vaiable amounts of water. The invention includes all such solvates.

The skilled artisan will further appreciate that certain compounds of the invention that exist in crystalline form, including the various solvates thereof, may exhibit polymorphism (i.e. the capacity to occur in different crystalline structures). These different crystalline forms are typically known as "polymorphs." The invention includes all such polymorphs. Polymorphs have the same chemical composition but differ in packing, geometrical arangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformability, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification. The skilled artisan will appreciate that different polymorphs may be produced, for example, by changing or adjusting the reaction conditions or reagents, used in making the compound. For example, changes in temperature, pressure, or solvent may result in polymorphs. In addition, one polymorph may spontaneously convert to another polymorph under certain conditions.

Compound Preparation

The compounds according to Formula I are prepared using conventional organic syntheses. Suitable synthetic routes are depicted below in the following general reaction scheme.

The skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. The protecting group may be removed at a suitable point in the reaction sequence to provide a desired intennediate or target compound. Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3rd ed.), John Wiley & Sons, NY (1999). In some instances, a substituent may be specifically selected to be reactive under the reaction conditions used. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful as an intermediate compound or is a desired substituent in a target compound.

Scheme 1

Formula I

[Conditions: a) bromine, EtOAc, 0 °C; then DMF, 85 °C; b) HOBt (or pyridine), EDC, acid, DC ]

Scheme 1 represents a general reaction scheme for preparing compounds of Formula I. Starting material substituted 1 ,3-propanediones 1.1 can be bromonated by bromine, which are coupled with thiourea 1.2 to give the amines 1.3. An appropriate acid can be reacted with the amines 1.3 in the presence of HOBt and EDC to obtain compounds of Formula I.

Examples

The following examples illustrate the invention. These examples are not intended to limit the scope of the present invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the present invention. While particular embodiments of the present invention are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention.

Abbreviations

BINAP 2,2 ' -bis (diphenylphosphino)- 1 , 1 ' -binaphthyl

DCM dichloromethane

DMF N,N-dimethylformamide

DMSO di methy lsulphoxi de

EA ethyl acetate

EDC N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride

ES Electrospray

HOBt Hydroxybenzotriazole

IPA isopropylalcohol

LCMS Liquid Chromatography Mass Spectrometry

MDAP mass directed automated preparative liquid chromatography.

MS mass spectrometry PE petroleum ether

PG protecting group

PMB p-methoxybenzyl

RT room temperature

sat. saturated

TFA trifluoroacetic acid

THF tetrahydroforan

Chromatography

Unless stated otherwise, all chromatography was carried out using silica columns.

LCMS Conditions:

1) Acidic conditions:

Mobile phase: water containing 0.05 % TFA / acetonitrile

Column: XBridgeTM CI S 30 x 100 mm - 5 microns

Detection: MS and photodiode array detector (PDA)

2) Basic conditions:

Mobile phase: water containing 0.08 % NH₄HCO₃ / acetonitrile

Column: XBridgeTM CI 8 30 x 100 mm - 5 microns;

Detection: MS and photodiode array detector (PDA)

MDAP Conditions:

1) Acidic conditions:

Instrument: Waters instrument

Column: Sunfire Prep CI 8 column (5 um, 19 x 50 mm)

Mobile phase: water containing 0.05% TFA / acetonitrile.

2) Basic conditions:

Instrumnet: Waters instrument

Column: Xbridge Prep CI S column (5 um, 19 x 50 mm)

Mobile phase: water containing 0.04% ammonia/ acetonitrile. Example 1

2-[4-(methylsulfonyl)phenylJ-A'-[4-phenyI-5-(phenylcarbonyl)-l,3-thiazol-2-yI]acetamide

Intermediate 1 a: (2-amino-4-phenylthiazol-5-yl)(phenyl)methanone

Bromine (4.59 mL) was added into a solution of l,3-diphenyl-l,3-propanedione (20 g) in ethyl acetate (50 mL) dropwise at 0 °C. After the addition was complete, the resultant mixture was stirred at 0 °C until the color of bromine faded away. Solvent was removed in vacuo. The residue and thiourea (6.79 g) was dissolved into N_tN-dimethylformamide (DMF) (20 mL) and the solution was heated to 85 °C for 15 mins. The mixture was cooled down, poured into water, basified by NaHC0 , and extracted with EtOAc (3 times). The combined organic layers were dried over anhydrous Na₂SO.(. After filtration, the filtrate was concentrated in vacuo. The solid formed was collected by filtration and washed with cold EtOAc, MeOH successively. The crude product was dried in vacuo to afford (2-amino -phenylthiazol-5-yl)(pheny!)methanone (14 g) as a yellow solid. The mother liquid was purified by chromatography on silica gel (EtOAc : PE = 0-50 %) to afford (2 -amino-4 -phenyl -1,3- ihiazol-5-yl)(phenyl)methanone (4 g) as a yellow solid. MS(ES⁺) m/z 281 (MH⁺).

Preparation of the final product

[4-(MethyIsulfonyl)phenyl]acetic acid (188 mg), (2-amino-4-phenyl-l ,3-thiazol-5- yl)(phenyl)methanone (intermediate la, 123 mg), EDC (202 mg) and HOBt (161 mg) were added into a 10 mL vial. Dichloromethane (DCM) (5 mL) was added. The reaction mixture was stirred at RT overnight. DCM was removed. The obtained crude was redissolved in DMF and submitted to MDAP for purification. Solvent was evaporated in vacuo to give the required product 2-[4- (methylsulfonyl)phenyl]-N-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2-yl]acetamide (155.5 mg) as a white powder. Ή-NMR (400 MHz, CDC1₃) 6 ppm 3.07 (s, 3H), 3.60 (s, 2H), 7.17-7.25 (m, 5H), 7.36-7.42 (m, 5H), 7.63 (d, J= 8.2 Hz, 2H), 7.94 (d, J= 8.3 Hz, 2H), 10.19 (br s, 1H); MS(ES⁺) m/z Ml (Ml-f ). Example 2

2-[4-(ethylsulfonyl)phenyI]-iV-[4- henyl-5-(phenylcarbonyl)-l,3-thiazol"2-yl]acetamide

Intermediate 2a: [4-(ethylsulfonyl)phenyHacetic acid

Step 1: A solution of sodium nitrite (18.4 g) in 133 mL of water was added dropwise at 0 °C, while stirring, to a suspension of (4-aminophenyl)acetic acid (40.2 g) in 133 mL of water and 54 mL of concentrated hydrochloric acid. After the addition was complete, the reaction mixture was stirred at the same temperature for 45 minutes. This solution of cold diazonium salt was then added dropwise at room temperature to a mixture of potassium ethylxanthate (49.4 g), 80 mL of water and 200 mL of 2 M sodium carbonate solution. The mixture was heated to 45 °C and stirred at this temperature until gas evolution stops. After cooling to room temperature, pH was adjusted to 1 with concentrated hydrochloric acid and the oiled xanthogenate ester was extracted with ether. Solvent was evaporated to give a dark red liquid (4-{[(ethyloxy)carbonothioyl]thio}phenyl)acetic acid (90 g). MS(ES⁺) m/∑ 257 (MH⁺).

Step 2: (4-{[(Ethyloxy)carbonothioyl]thio}phenyl)acetic acid (90 g) was taken up in 340 mL of ethanol, and a solution of 70 g of potassium hydroxide in 340 mL of water was added. Boiling at reflux was effected for 20 hours. The major portion of ethanol was subsquent!y removed by the distillation under reduced pressure. The aqueous phase was cooled with ice, and acidified with concentrated hydrochloric acid while stirring. The obtained solution was extracted with diethyl ether (500 mL), The organic phase was washed with brine, dried over anhydrous sodium sulphate, filtered, and concentrated to affored the desired product as a yellow solid (4-mercaptophenyl) acetic acid (33 g). MS(ES⁺) m/z 169 (MH⁺).

Step 3: To a solution of (4-mercaptophenyl) acetic acid (33 g) in NN-dimethylformamide (DMF) (240 mL) was added K₂C0₃ (108 g) and bromoethane (64.1 g). The reaction mixture was stirred at RT. After 2,5 hours, the starting material was totally consumed. The reaction mixture was partitioned between ethyl acetate (300 mL) and water (300 mL). The organic phase was washed with water (300 mL x 4) and brine (200 mL), dried over sodium sulphate, filtered, and concentrated to give the desired product ethyl [4-(ethylthio)phenyl]acetate (34 g) as a pale yellow solid. MS(ES⁺) m/z 225 (MH⁺).

Step 4: A solution of ethyl [4 -(ethyl thio)phenyI] acetate (34 g) in dichloromethane (DCM) (500 mL) was cooled to 0 °C with an ice bath. MCPBA (78 g) was added in portions, and the reaction mixture was stirred at RT overnight. The obtained suspension was filtered. The filtrate was washed with sat. sodium carbonate solution (400 mL x 2), water (500 mL), then brine (250 mL). The obtained solution was dried over sodium sulphate, filtered, and concentrated. The residue was purified by column chromatography (silica gel; EtOAc : PE = 0: 1 to 1 : 1) to afford the target compound ethyl [4-(ethy!sulfonyl)phenyl]acetate as a yellow liquid (25 g). MS(ES⁺) m/z 257 (MH⁺).

Step 5: To a solution of ethyl [4-(ethylsulfonyl)phenyl] acetate (25 g) in ethanol (180 mL) was added a solution of NaOH (14.28 g) in water (180 mL). The reaction mixture was stirred at room temperature overnight. Ethanol was removed under reduced pressure, and 150 ml of water was added. The aqueous phase was washed with dichloromethane (100 mL x 2), and then acidified with 6 M HC1 to pH = 1. This solution was extracted with ethyl acetate (200 mL x 2). The combined organic phases were washed with brine (200 mL), dried over sodium sulphate, filtered, and concentrated to give the desired product as a dark red oil, which slowly solidified to give a yellow solid [4- (ethylsulfonyl)phenyl]acetic acid (20 g). Ή-NMR (400 MHz, DMSO-<¾) δ ppm 1.07 (t, J - 9.6 Hz, 3H), 3.26 (q, J= 9.6 Hz, 2H), 3.72 (s, 2H), 7.53 (d, 7= 11.2 Hz, 2H), 7.81 (d, J= 1 1.2 Hz, 2H), 12.53 (s, 1H); MS(ES⁺) m/z 229 (MH⁺).

Preparation of the final product

[4-(Ethylsulfonyl)phenyl] acetic acid (intermediate 2a, 100 mg), (2-amino-4-phenyl-l ,3- thiazoI-5-yl)(phenyl)methanone (intermediate la, 123 mg), EDC (101 mg) and HOBt (81 mg) were added into a 10 mL vial. Dichloromethane (DCM) (5 mL) was added. The reaction mixture was stirred at RT overnight. DCM was removed. The obtained crude was redissolved in DMF and submitted to MDAP for purification. Solvent was evaporated in vacuo to give the required product 2- [4-(ethylsulfonyl)phenyl]-N-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2-yl]acetamide (158 mg) as a white powder. ^!H-NMR (400 MHz, CD ¾) δ ppm 1.31 (t, /= 7.4 Hz, 3H), 3.13 (q, J = 7.4 Hz, 2H), 3.55-3.59 (m, 2H), 7.16-7.25 (m, 5H), 7.36-7.43 (m, 5H), 7.64 (d, J= 8.4 Hz, 2H), 7.89 (d, J= 8.2 Hz, 2H), 10.31 (br s, 1H); MS(ES⁺) m/z A9\ (MH⁺).

Example 3

2-[4-(ethyIsulfonyI)phenyl]-2-methyl-yV-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2- yljpropanamide

Intermediate 3a: 2-f4-iethylsulfonynphenyl1-2-methylpropanoie acid

Step 1: To a suspension of [4-(ethylsulfonyl)phenyl] acetic acid (intermediate 2a, 203.1 mg) and cesium carbonate (1565 mg) in N,N-dimethylformamide (DMF) (7 mL) was added Mel (0.334 mL). The reaction mixture was stirred at RT overnight. The reaction mixture was diluted with water, and acidified with 3 M HC1 to pH ~ 3. The resulting mixture was partitioned between EtOAc and water. The organic phase was dried over Na2S0₄, filtered, and concentrated to afford the crude product which was purified by chromatography (EtOAc : PE = 0: 1 to 1 :4) to afford the intermediate methyl 2-[4-(ethylsulfonyl)phenyI]-2-methylpropanoate.

Step 2: The intermediate methyl 2-[4-(ethylsulfonyl)phenyl]-2-methylpropanoate was dissolved in 10 mL EtOH and 10 mL 2 M NaOH solution. The obtained mixture was stirred at RT for 3 hours. 3 M HC1 was added to adjust pH to ~ 3. The resulting mixture was partitioned between EtOAc and water. The organic phase was dried over N ₂S0₄. Filtration and evaporation afforded 2- [4-(ethylsu!fonyl)pheny!]-2-methylpropanoic acid (200 mg). MS(ES⁺) mfz 257 (MH⁺).

Preparation of the final product 2-[4-(Ethylsulfonyl)phenyl]-2-metliylpropanoic acid (intermediate 3a, 67.1 mg), (2-amino-4- phenyl-l ,3-thiazol-5-yl)(phenyl)methanone (intermediate la, 81 mg), EDC (65.2 mg) and HOBt (52.1 mg) were added into a 10 mL vial. Dichloromethane (DCM) (10 mL) was added. The reaction mixture was stirred at RT. After 3 days, DCM was removed. The obtained mixture was redissolved in DMF/THF and purified by MDAP. Solvent was freeze-dried to give the required 2-[4-

(ethylsulfonyl)phenyl] -2-methyl -N- [4-phenyl-5 - (phenyl carb onyl)- 1 ,3-thiazol -2-yl ] propanamide (1 10.2 mg) as a white solid. Ή-NMR (400 MHz, DMSO-<¾ 6 ppm 1.04 (t, J= 7.3 Hz, 3H), 1.60 (s, 6H), 3.23 (q, J= 7.3 Hz, 2H), 7.05-7.15 (m, 3H), 7.18 (t, J = 7.7 Hz, 2H), 7.26 (d, J - 6.8 Hz, 2H), 7.36 (t, J= 7.4 Hz, 1H), 7.49 (d, J= 7.2 Hz, 2H), 7.53 (d, J= 8.5 Hz, 2H), 7.83 (d, J = 8.5 Hz, 2H), 12.42 (s, 1H); MS(ES⁺) m/z 519 (MH⁺).

Example 4

2-[4-(ethyIsulfonyI)phenyl]-JV-[4- henyl-5-(phenylcarbonyl)-l,3-thiazol-2-yl]propanamide

Intermediate 4a: 2-(4-(ethylsulfonyl)phenyl)propanoic acid

Step 1: To a solution of [4-(ethylsulfonyl)phenyl]acetic acid (intermediate 2a, 760 mg) in methanol (5 mL) was added SOC½ (0.267 mL) at RT, The reaction mixture was heated to reflux (oil bath temperature: 80 °C) and stirred overnight. Solvent was removed under vacuum to afford methyl [4-(ethylsulfonyl)phenyl]acetate (790 mg) as a light yellow oil. MS(ES⁺) m/z 243 (MH⁺).

Step 2: To a solution of methyl [4-(ethylsulfonyl)phenyl]acetate (303 mg) in tetrahydrofuran

(THF) (1 mL) was added sodium hydride (75 mg) slowly. The vial was sealed and stirred at RT. After 30 mins, a solution of methyl iodide (0.070 mL) in tetrahydrofuran (THF) (1 mL) was carefully addeded at 0 °C. After stirring at this temperature for 3 hours, the reaction mixture was quenched with water. 1 M HCl was used to adjust pH to about 5. The organic phase was extracted with EtOAc for 3 times. The combined organics were dried over NaaSCv After removal of solvent, the obtained crude was purified by chromatography eluting with EtOAc : PE = 0: 1 to 1 :2 to afford the intermediate methyl 2-[4-(ethylsulfonyl)phenyl]propanoate. MS(ES^÷) m/z 257 (MH^÷).

Step 3: The intennediate methyl 2-[4-(ethylsulfonyl)phenyl]propanoate was dissolved in EtOH (2 mL). 2 M NaOH (2 mL) was added. The mixture was stirred at RT for 2 hours. 1 M HC1 was used to adjust pH to about 5. The organic phase was extracted with EtOAc for 3 times. The combined organics were dried over Na^SO^ After removal of solvent, 2-(4-(ethylsulfonyl)phenyI)propanoic acid (202.7 mg) was obtained and used directly in the next step. MS(ES⁺) m/z 243 (MH⁺).

Preparation of the final product

2-[4-(EthyIsulfonyI)phenyl]propanoic acid (intermediate 4a, 202 mg), (2-amino-4-phenyl-l ,3- thiazol-5-yl)(phenyl)methanone (intennediate la, 234 mg), EDC (192 mg) and HOBt (153 mg) were added into a 10 mL vial. Dichloromethane (DCM) (5 mL) was added. The reaction mixture was stirred at RT overnight. DCM was removed. The obtained crude was redissolved in DMF and submitted to MDAP for purification. Solvent was evaporated in vacuo to give the required product 2» [4-(ethylsulfonyl)phenyl]-N-[4-phenyl-5-(phenylcarbonyl)- 1 ,3-thiazol-2-yl]propanamide (88.6 mg) as a yellowish solid. 'H-NMR (400 MHz, DMSCkfc) δ ppm 1.03 (t, J= 7.4 Hz, 3H), 1.44 (d, J= 7.0 Hz, 3H), 3.21 (q, J= 7.3 Hz, 2H), 4.08 (d, J= 6.8 Hz, 1H), 7.08-7.20 (m, 5H), 7.24-7.31 (m, 2H), 7.31- 7.38 (m, 1H), 7.47 (d, J= 6.8 Hz, 2H), 7.60 (d, J= 8.3 Hz, 2H), 7.81 (d, J = 8.5 Hz, 2H), 13.02 (br s, 1H); MS(ES⁺) m/z 505 (MET).

Example 5

2-{4-[(methylamino)sulfonyl]phenyl}-A^r-[4-phenyl-5-(phenykarbonyl)-l,3-thiazol-2- yl]acetamide

Intennediate 5a: 2-f4-(N-methyisulfamoyl^'>phenyl^')acetic acid

Step 1 : Ch!orosulfonic acid (70 mL) was added into phenylacetic acid (10 g) dropwise at 0 °C. The reaction mixture was stirred at 0 °C for 1 hour, and then warmed to room temperature and stirred overnight. The reaction mixture was poured onto ice and left until all ice melted. The suspension was filtered. The obtained solid was washed with cold water and dried in vacuo to afford a white solid (10 g), which contains isomers. The mixture was recrystallized from chloroform twice to give the title product 2-(4-(chlorosulfonyl)phenyl)acetic acid (2.2 g). Ή-NM (400 MHz, DMSO- ) 5 ppm 3.57 (s, 2H), 7.21 (d, J= 8.0 Hz, 2H), 7.54 (d, J= 8.0 Hz, 2H), 10.52 (s, 1H); MS(ES⁺) m/z 235 (MH⁺);

Step 2: 2-(4-(Chlorosulfonyl)phenyl)acetic acid (2 g) was added to a solution of methylamine in methanol (0.265 g). The reaction mixture was stirred at room temperature for 1 hour. NaOH (0.5 g) was added to the reaction mixture to release the amine from its HCI salt. Solvent was removed under reduced pressure. The residue was dissolved in water (20 mL), acidified to pH = 1 with 6 M HCI solution, and extracted with dichloromethane (50 mL x 3). The organic phases were combined, dried over anhydrous sodium sulphate, and concentrated to give a white solid (0.8 g). The aqueous phase still contained the product: Solvent was removed, and THF was added to extract the product. The organic phase was dried and concentrated to give a yellow solid (0.8 g). The solid products were combined, to which diethyl ether (20 mL) was added and the suspension was sonicated for 20 mins. The solid was collected by filtration, washed with ether, and dried in vacuo to give the title compound 2-(4-(N-methylsulfamoyl)phenyl)acetic acid (1.2 g) as a white solid. Ή-NMR (400 MHz, DMSO-de) δ ppm 2.41 (d, J= 4.8 Hz, 3H), 3.70 (s, 2H), 7.41 (q, J= 4.8 Hz, 2H), 7.49 (d, J= 8.0 Hz 1H), 7.72 (d, J= 8.0 Hz, 2H), 12.50 (br s, 1H); MS(ES⁺) m/z 230 (MH⁺).

Preparation of the final product

A mixture of (2-amino-4-phenyl-l ,3-thiazol-5-yl)(phenyI)methanone (intermediate la, 70 mg), {4-[(methylamino)sulfonyl]phenyl}acetic acid (intermediate 5a, 63.0 mg), HOBt (50.6 mg) and EDC (71.8 mg) in dichloromethane (DCM) (3 mL) was stirred at room temperature under N2 overnight. Solvent was removed under reduced pressure. The residue was purified by MDAP directly to afford 2- {4-[(methylamino)sulfonyl]phenyI} -N-[4-phenyl-5-(phenylcarbonyl)-l ,3-thiazol-2- y]]acetamide (62 mg) as a white solid. Ή-NMR (400 MHz, DMSO-<¾) δ ppm 2.41 (d, 7- 4.8 Hz, 3H), 3.98 (s, 2H), 7.17-7.27 (m, 5H), 7.36-7.38 (m, 2H), 7.41-7.47 (m, 2H), 7.54-7.59 (m, 4H), 7.76 (d, J= 8.4 Hz, 2H), 13.06 (s, 1H); MS(ES⁺) m/z 492 (MH⁺).

Example 6

2-{4-[(ethylamino)sulfonyl]phenyl}- V-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazoI-2-yl]acetamide

Intermediate 6a: 2-(^'4-fN-methylsulfamQyl^')phenyl)acetic acid

Step 1 : see step 1 for preparing intermediate 5a.

Step 2: 2-(4-(Chlorosulfonyl)pheny!)acetic acid (2.2 g) was added to a solution of ethylamine in methanol (30 %, 80 mL) cooled at 0 °C, and the reaction mixture was stirred at room temperature for 1 h. Sodium hydroxide (0.5 g) was added to release the ethylamine from its salt. Solvent (together with excess ethylamine) was removed in vacuo. The residue was dissolved in water (20 mL), and the aqueous solution was acidified with 6 M HC1 to pH = 1. The oily product was extracted with dichloromethane (50 mL x 3), and the combined organics were dried over anhydrous sodium sulphate, filtered, and concentrated to give the desired product 2-(4-(N-methyisulfamoyl)phenyl)acetic acid (2.0 g) as a white solid. Ή-NMR (400 MHz, DMSO-ifc) δ ppm 0.97 (t, J= 7.2 Hz, 3H), 2.77 (m, 2H), 3.70 (s, 2H), 7.47 (d, J = 8.0 Hz, 2H), 7.52 (t, J= 5.6 Hz, 1H), 7.73 (d, J = 8.0 Hz, 2H), 12.49 (s, 1 H); MS(ES⁺) m/z 244 (MH⁺).

Preparation of the final product

A mixture of {4-[(ethylamino)sulfonyl]phenyl}acetic acid (intermediate 6a, 66.8 mg), EDC (62.2 mg) and HOBt (43.9 mg) in dichloromethane (DCM) (2 mL) was stirred at room temperature under nitrogen for 10 mins. Then (2-amino-4-pheny!- l ,3-thiazol-5-yl)(phenyl)methanone

(intermediate l a, 70 mg) was added, and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was partitioned between DCM and water. The aqueous phase was extracted with DCM for 3 times. The combined organic layers were washed with brine and dried over anhydrous Na₂S0 . After filtration, solvent was removed under reduced pressure and the residue was purified by MDAP to afford 2- {4-[(ethylamino)sulfonyl]phenyl}-N-[4-phenyl-5- (phenylcarbonyl)-l,3-thiazol-2-yl]acetamide (70 mg) as a white solid. 'H-NMR (400 MHz, DMSO-i¾) δ ppm 0.97 (t, J= 7.2 Hz, 3H), 2.50-2.81 (m, 2H), 3.98 (s, 2H), 7.17-7.27 (m, 5H), 7.36-7.38 (m, 2H), 7.43 (d, J= 7.6 Hz, 1H), 7.54-7.57 (m, 5H), 7.77 (d, J= 8.4 Hz, 2H), 13.06 (s, 1H); MS(ES⁺) m/z 506 (MH*).

Example 7

/V-I4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2-yl]-2-{4-[(2,2,2- triflxioroethyl)sulfonyl]phenyl}acetamide

Step 1 to 2: see steps 1 &2 for preparing intermediate 2a.

Step 3: A mixture of 2-(4-rnercaptophenyl)acetic acid (300 mg) and H₂S0 (0.01 mL) in methanol (10 mL) was heated to 70 °C with stirring for 45 mins. The reaction mixture was cooled to 0 °C in an ice bath. The solution was neutralized with NaHC(¾ to pH = 7, and then concentrated to 5 mL. DCM (15 mL) and water (10 mL) was added. The aqueous was extracted with DCM (10 mL x 3). The combined organic layers were dried over Na₂S0₄, filtered, and concentrated to give the crude product (300 mg). MS(ES⁺) m/z 183 (MH⁺).

Step 4: A mixture of methyl 2-(4-mercaptophenyl)acetate (1.2 g), l , l,l-trifluoro-2-iodoethane (1.4 g) and K₂C0₃ (2.4 g) in N,N-dimethy!formamide (DMF) (30 mL) was stirred at room temperature for 3 hours. Water (50 mL) and DCM (50mL) was added. The aqueous was extracted with DCM (30 mL x 4), The combined organic phases were washed with brine (30 mL x 3), dried over Na₂S(¾, filtered, and concentrated to give the crude product (1.2 g). MS(ES⁺) m/z 265 (MH⁺).

Step 5: A mixture of methyl 2-(4-mercaptophenyl)acetate (1.2 g) and mCPBA (1.9 g) in dichloromethane (DCM) (50 mL) was stirred at room temperature overnight. Solvent was evaporated under reduced pressure. DCM (30 mL) was added and the mixture was filtered. The filtrate was concentrated, and the residue was purified by column chromatography (silica gel; PE : EA = 100:0 to 100:3) to afford methyl 2-(4-(2,2,2-trifluoroethylthio)phenyl)acetate (1.2 g) as an oil. MS(ES⁺) m/z 297 (MH⁺).

Step 6: A mixture of methyl 2-(4-(2,2,2-trifluoroethyIsulfonyl)phenyl)acetate (1.1 g) and cone.

HC1 (30 mL) in acetic acid (30 mL) was stirred at 105 °C for 1.5 hours. Solvent was removed under reduce pressure. Water (30 mL) and DCM (30 mL) was added. The aqueous was washed with DCM (15 mL x 3). The combined organic phases were dried over Na₂S0 , filtered, and concentrated to give the title product 2-(4-(2,2,2-trifluoroethylsulfonyl)phenyl)acetic acid (700 mg) as a white solid. Ή-NMR (400 MHz, DMSO-tf₆) δ ppm 3,77 (s, 2H), 4.94 (q, J = 10.0 Hz, 2H), 7.59 (d, J= 8.0 Hz, 2H), 7.91 (d, J= 8.0 Hz, 2H), 12.57 (s, 1H); MS(ES⁺) m/z 283 (MH⁺).

Preparation of the final product

A mixture of (2-amino-4-phenyl-l ,3-thiazol-5-yl)(phenyl)methanone (intermediate la, 70 mg), {4-[(2,2,2-trifluoroethyl)sulfonyl]phenyl} acetic acid (intermediate 7a, 74.0 mg), HOBt (50.6 mg) and EDC (71.8 mg) in dichloromethane (DCM) (2 mL) was stirred at room temperature overnight under N₂. Solvent was removed under reduced pressure, and the residue was purified by MDAP to afford N-[4-phenyl-5-(phenylcarbonyl)-l ,3-thiazol-2-yl]-2-{4-[(2,2,2- trifluoroethyl)sulfonyl]phenyl}acetamide (46 mg) as a white solid. Ή-NMR (400 MHz, DMSO-i ) 5 ppm 4.04 (s, 2H), 4.96 (q, J= 10,0 Hz, 2H), 7,19-7.27 (m, 5H), 7.39-7.36 (m, 2H), 7.45-7.41 (m, 1 H), 7.53-7.56 (m, 2H), 7.67 (d, J= 8.4 Hz, 2H), 7.95 (d, J- 8.4 Hz, 2H), 13.08 (s, 1H); ¹⁹F-NMR (376 MHz, DMSO-ί/ΰ) δ ppm -59.79; MS(ES⁺) m/z 545 (MH⁺).

Example 8

iV-[4-phenyl-S-(phenyIcarbon l)-l,3-thiazol-2-yl]-2-[4-(propylsulfonyl)phenyl]acetamide

Intermediate 8a: 2-(4-(propylsulfonyi)phenyl)acetic acid

Step 1 and 2: see steps 1&2 for preparing intermediate 2a.

Step 3: To a solution of (4-mercaptophenyl)acetic acid (18 g) in N,N-dimethylforrnamide (DMF) (240 mL) was added K₂C0₃ (59.2 g) and 1-iodopropane (54.6 g). The reaction mixture was stirred at RT for 2.5 hours. The reaction mixture was partitioned between ethyl acetate (300 mL) and water (300 mL), The organic layer was washed with water (300 mL x 4) and brine (200 mL), dried with sodium sulphate, filtered, and concentrated to give the desired product propyl [4- (propylthio)phenyl] acetate (23 g) as a pale yellow solid. MS(ES^÷) m/z 253 (MH⁺).

Step 4: Propyl [4-(propylthio)phenyl]acetate (23 g) was dissolved in dichloromethane (DCM) (500 mL), and the solution was cooled to 0 °C with an ice bath. MCPBA (15.73 g) was added in portions, and the reaction mixture was stirred at RT overnight. The reaction mixture was filtered to removed the solid. The filtrate was washed with sat. sodium carbonate solution (400 mL x 2), water (500 mL), brine (250 mL), dried over sodium sulphate, filtered, and concentrated. The residue was purified by column chromatography (silica gel; ethyl acetate : petroleum = 0: 1 to 1 : 1) to afford the target compound propyl [4-(propylsulfonyl)phenyl]acetate (12 g) as a white soild. MS(ES ) m/z 285 (ΜΙ-Γ).

Step 5: To a solution of propyl [4-(propylsulfonyl)phenyl]acetate (12 g) in ethanol (180 mL) was added a solution of NaOH (5.06 g) in water (180 mL). The reaction mixture was stirred at room temperature overnight. Ethanol was removed under reduced pressure, and 150 mL of water was added to the aqueous solution. The solution was washed with dichloromethane (100 mL x 2), and then acidified with 6 HC1 to pH = 1. The obtained solution was extracted with ethyl acetate (200 mL x 2). The combined organic phases were washed with brine (200 mL), dried over sodium sulphate, filtered, and concentrated to give the desired product 2-(4-(propylsulfonyl)phenyl)acetic acid (7 g) as a whie soild. ^lH-NMR (400 MHz, DMSCW₆) δ ppm 1.00 (t, J= 7.2 Hz, 3H), 1.74 (m,

2H), 3.06 (t, J= 8.0 Hz, 2H), 3.76 (s, 2H), 7.49 (d, J = 8.4 Hz, 2H), 7.88 (d, J = 8.4 Hz, 2H); MS(ES⁺) m/z 243 (MH⁺).

Preparation of the final product

A mixture of (2-amino-4-phenyl-l,3-thiazol-5-yl)(phenyl)methanone (intermediate la, 70 mg), [4-(propylsulfonyl)phenyI]acetic acid (intermediate 8a, 63.5 mg), EDC (62.2 mg) and HOBt (43.9 mg) in dichloromethane (DCM) (3.5 mL) was stirred at room temperature overnight. Solvent was removed under reduced pressure. The residue was purified by MDAP to afford N-[4-phenyl-5- (phenylcarbonyl)-l ,3-thiazol-2-yl]-2-[4-(propylsulfonyl)phenyl]acetamide (61 mg) as a white solid. Ή-NMR (400 MHz, DMSO-d₆) δ ppm 0.91 (t, J= 7.4 Hz, 3H), 1.50-1.60 (m, 2H), 3.25-3.29 (m, 2H), 4.02 (s, 2H), 7.17-7.27 (m, 5H), 7.37-7.39 (m, 2H), 7.43 (t, J= 7.4 Hz, 1H), 7.54-7.56 (m, 2H), 7.63 (d, J= 8.3 Hz, 2H), 7.88 (d, J= 8.3 Hz, 2H), 13.08 (s, 1H); MS(ES⁺) m/z 505 (MH⁺).

Example 9

2-(4-{[2-(methyloxy)ethyl]sulfonyl}phenyI)-N-[4-phenyl-5-(phenyIcarbonyl)-l^-thiazoI-2- yljacetamide

Intermediate 9a: 2-(4-((2-memoxyetlwl)sulfinyl pbenyl)acetic acid

Step 1: A mixture of 2-(4-mercaptophenyl)acetic acid (3.0 g, synthesis of this starting material, see steps 1 &2 for preparing intermediate 2a) and H₂S0 (0.01 mL) in methanol (10 mL) was heated to 70 °C with stirring for 45 mins. The reaction mixture was cooled to 0 °C in ice bath, and then neutralized with NaHCOj to pH = 7. Solvent was concentrated to about 5 mL. Then DCM (15 mL) and water (10 mL) was added. The aqueous phase was extracted with DCM (10 mL x 3). The combined organics were dried over Na₂SC>4, filtered, and concentrated to give the crude product (2.8 g). MS(ES⁺) m/ l 83 (MH⁺).

Step 2: A mixture of methyl 2-(4-mercaptophenyl)acetate (2.8 g), l -bromo-2-methoxyethane (2.136 g) and K₂C0₃ (5.31 g) in NN-dimethylformamide (DMF) (50 mL) was stirred at room temperature for 3 hours. Water (15 mL) and DCM (20mL) was added, and the aqueous was extracted with DCM (15 mL x 4). The combined organic phases were washed with brine (20 mL x 3), dried over Na₂S0₄, filtered, and concentrated to oily product (2.6 g). MS(ES⁺) m/z 241 (MH*).

Step 3: A mixture of methyl 2-(4-(2-methoxyethylthio)phenyI)acetate (2.6 g) and mCPBA (1.867 g) in dichloromethane (DCM) (20 mL) was stirred at room temperature for 3 hours. Water (15 mL) and DCM (20 mL) was added. The aqueous was extracted with DCM (15 mL x 4). The combined organic phases were washed with 1 M NaOH (20 mL x 4), brine (20 mL x 2), dried over Na₂S04, filtered, and concentrated. The crude product was purified by column chromatography (silica gel; EA : PE = 50: 1 to 8: 1) to give the desired methyl 2-(4-(2-methoxyethylsulfony]) phenyl)acetate as an oil (2.3 g). MS(ES⁺) m/z 273 (MH⁺).

Step 4: A mixture of methyl 2-(4-(2-methoxyethylthio)phenyl)acetate (2.3 g) and OH (1.176 g) in methanol (15 mL) and water (15 mL) was stirred at room temperature overnight. Water (1 mL) and DCM (20 mL) was added, and the aqueous phase was extracted with DCM (15 mL x 4). The combined organic phases were washed with brine (20 mL x 3), dried over Na₂SC>4, filtered, and concentrated. The crude product was purified by column chromatography (silica gel; DCM : MeOH = 100:0 to 50: 1) to give the desired product 2-(4-(2-methoxyethyl sulfonyl) phenyl)acetic acid (788 mg) as a white solid. Ή-NMR (400 MHz, DMSO-c¾ δ ppm 3.11 (s, 3H), 3.60 (m, 4H), 3.74 (s, 2H), 7.53 (d, J= 8.4 Hz, 2H), 7.84 (d, J= 8.4 Hz, 2H), 12.54 (s, 1H); MSfES⁴) m/z 259 (MH⁺), Preparation of the final product

A mixture of (2-araino-4-phenyl-l ,3-thiazol-5-yl)(phenyI)methanone (intermediate l a, 80 mg), (4-{[2-(methyloxy)ethyl]sulfonyl}phenyl)acetic acid (intermediate 9a, 77 mg), EDC (71.1 mg) and HOBt (50.1 mg) in dichloromethane (DCM) (3.5 mL) was stirred at room temperature overnight. Solvent was removed under reduced pressure and the residue was purified by MDAP to afford 2-(4- {[2-(methyloxy)ethyl]sulfonyl}phenyl)-N-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2-yl]acetamide (63 mg) as a white solid. Ή- MR (400 MHz, DMSO-i¾) δ ppm 3.10 (s, 3H), 3.57-3.63 (m, 4H), 4.01 (s, 2H), 7.17-7.27 (m, 5H), 7.36-7.38 (m, 2H), 7.43 (t, J= 7.4 Hz, 1H), 7.54-7.56 (m, 2H), 7.61 (d, J = 8.3 Hz, 2H), 7.88 (d, J = 8.3 Hz, 2H), 13.07 (s, 1 H); MS(ES⁺) m/z 521 (MH⁺). Example 10

2-{4-[{2-hydroxyethyl)sulfonyl]phenyl}-iV-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2- yl]acetamide

Intermediate 10a: (4-[^"(^'2-hvdroxyethyl^')suIfonyllphenvUacetic acid

Step 1 : see step 1 for preparing intermediate 9a.

Step 2: A mixture of methyl 2-(4-mercaptophenyl)acetate (3.5 g), 2-bromoethanol (1.716 g) and Cs₂C0₃ (6.10 g) in N,N-dimethylformamide (DMF) (50 mL) was stirred at room temperature overnight. The mixture was filtered, and DCM (50 mL) was added. The solution was washed with water (30 mL x 5), brine (30 mL x 2), dried over Na₂S0₄, filtered, and concentrated. The crude product was purified by flash chromatography (silica gel; PE : EA = 20: 1 to 2: 1) to give the desired product (3.0 g) as a white solid. MS(ES⁺) m/z 227 (MH⁺).

Step 3: A mixture of methyl 2-(4-(2-hydroxyethyHhio)phenyl)acetate (3.0 g) and mCPBA (3.89 g) in dichloromethane (DCM) (30 mL) cooled in an ice bath was stirred for 14 hours. 1 M NaOH (20 mL) was added, and the organic phase was washed with brine (15 mL x 3), dried over Na₂SC>4, filtered, and concentrated. The crude product was purified by column chromatography (silica gel; PE : EA = 10: 1 to 1 : 1) to afford the desired product (1.2 g) as a white solid. MS(ES⁺) m/z 259 (MH⁺).

Step 4: Lithium hydroxide monohydrate (0.334 g) in water (50 mL) was added into a solution of methyl 2-(4-(2-hydroxyethylsulfonyl)phenyI)acetate (1.2 g) in tetrahydrofuran (THF) (50.0 mL). The reaction mixture was stirred at room temperature overnight. Solvent was removed under reduced pressure, and the mixture was cooled in an ice bath and acidified to pH = 1-2 with cone. HC1. The aqueous phase was extracted with EtOAc (30 mL x 5). The combined organic phases were washed with brine (30 mL x 2), dried over Na₂S0₄, filtered, and concentrated to give the desired product (800 mg) as a white solid. Ή-NMR (400 MHz, DMSO-i ₆) 6 ppm 3.44 (t, J= 5.6 Hz, 2H), 3.56 (t, J = 5.6 Hz 2H), 3.74 (s, 2H), 4.83 (br s, 1H), 7.53 (d, J= 8.0 Hz, 2H), 7.84 (d, J= 8.0 Hz, 2H), 12.55 (s, 1H); MS(ES⁺) m/z 245 (MH⁺).

Preparation of the final product

A mixture of (2-amino-4-phenyl-l,3-thiazol-5-yl)(phenyl)methanone (intermediate la, 70 mg), {4 -[(2-hydrox ethyl )sulfonyl]phenyl} acetic acid (intermediate 10a, 67.1 mg), HOBt (50.6 mg) and EDC (71.8 mg) in dichloromethane (DCM) (2 mL) was stirred at room temperature under N₂ overnight. Solvent was removed under reduced pressure. The residue was purified by MDAP to afford 2-{4-[(2-hydroxyethyl)sulfonyl]phenyl}-N-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazoI-2- yljacetamide (25 mg) as a white solid. Ή-NMR (400 MHz, DMSO-i/₆) δ ppm 3.45 (t, J = 6.4 Hz, 2H), 3.67 (t, J= 5.6 Hz, 2H), 4.01 (s, 2H), 4.84-4.97 (m, 1H), 7.17-7.28 (m, 5H), 7.36-7.38 (m, 2H), 7.43 (t, J= 3.2 Hz, 1H), 7.54-7.56 (m, 2H), 7.61 (d, J= 8.4 Hz, 2H), 7.88 (d, J= 8.4 Hz, 2H), 13.08 (s, 1H); MS(ES⁺) m/z 507 (MH⁺).

Example 11

2-[2-(ethylsulfonyl)phenyl]-iV-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2-yl]acetamide

Intermediate 11a: r2-amino-4-(3-chlorophenyn-1.3-thiazol-5-yl](2-chlorophenyl)metlianone

Step 1: To a suspension of indolin-2-one (5 g) in water (50 mL) was added NaOH (6.01 g). The mixture was stirred overnight at 100 °C. The pH was adjusted to 1 with concentrated hydrochloric acid to give the solution of 2-(2-aminophenyl)acetic acid in water. S(ES⁺) m/z 152 (ΜΙ-Γ).

Step 2: A solution of sodium nitrite (2.56 g) in 20 mL of water was added dropwise to a suspension of the above 2-(2-aminophenyl)acetic acid in 20 mL of water and 2.7 mL of concentrated hydrochloric acid cooled at 0 °C. After the addition was complete, the reaction mixture was stirred at the same temperature for a further 45 minutes. This cold diazonium salt solution was then added dropwise at room temperature to a mixture of potassium O-ethyl carbonodithioate (6.88 g), 20 mL of water and 28 mL of a 2 M sodium carbonate solution, and heating was effected to 45 °C until gas evolution stops. The mixture was subsequently cooled to room temperature, and the pH was adjusted to 1 with concentrated hydrochloric acid. The oiled xanthogenate ester was extracted with ether. Removal of solvent gave a dark red liquid 2-(2-(ethoxycarbonothioylthio)phenyl)acetic acid (9.5 g). MS(ES⁺) m/z 257 (MH⁺).

Step 3: 2-(2-(ethoxycarbonothioylthio)phenyl)acetic acid (9.5 g) was taken up in 40 mL of ethanol, and a solution of KOH (6.24 g) in 30 mL of water was added and boiling at reflux was effected for 20 hours. The major portion of ethanol was subsequently removed by the distillation at reduced pressure. The aqueous phase was cooled with ice, and was rendered acid with concentrated hydrochloric acid while stirring well. The solution was extracted with diethyl ether (100 mL x 5). The organic phase was washed with brine, dried over anhydrous sodium sulphate, filtered, and concentrated to afford the desired product 2-(2-mercaptophenyl)acetic acid (6 g). MS(ES⁺) m/z 169 (MH⁺).

Step 4: To a solution of 2-(2-mercaptophenyl)acetic acid (6 g) in N,N-dimethylformamide (D F) (80 mL) was added K₂C0₃ (34.5 g) and bromoethane (1 .43 g). The reaction mixture was stirred at 30 °C overnight. The reaction mixture was partitioned between ethyl acetate (300 mL) and water (100 mL). The organic phase was washed with water (100 mL x 4), brine (200 mL), dried over sodium sulfate, filtered, and concentrated to give the desired product ethyl 2-{2- (ethylthio)phenyl)acetate (6 g) as a brown solid. ¹ H-NMR (400 MHz, DMSO-c/₆) δ ppm 1 , 16 (t, J = 9,6 Hz, 3H), 2.91 (q, J= 9.6 Hz, 2H), 3.78 (s, 2H), 4.08 (q, J= 9.6 Hz, 2H), 7.16-7.21 (m, 1H), 7.26- 7.31 (m, 2H), 7.39-7.42 (m, 1H); MS(ES⁺) m/z 225 (MH⁺).

Step 5: Ethyl 2-(2-(ethylthio)phenyl)acetate (6 g) was dissolved in dichloromethane (DCM) (50 mL), and the solution was cooled to 0 °C with an ice bath. MCPBA (13,85 g) was added in portions, and the reaction mixture was stirred at room temperature overnight. The reaction mixture was filtered to remove the solid. The filtrate was washed with sat. sodium carbonate solution (50 mL x 2), water (50 mL), brine (150 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel; EtOAc / PE = 1 :6) to afford the target compound ethyl 2-(2-(ethylsulfonyl)phenyl)acetate (2.5 g) as a yellow solid. ^lH-NMR (400 MHz, DMSO-i/₆) δ ppm 1.10 (t, J= 7.2 Hz, 3H), 1.18 (t, J= 7.2 Hz, 3H), 3.27 (q, J= 7.2 Hz, 2H), 4.09 (q, J= 7.2 Hz, 2H), 4.12 (s, 2H), 7.53 (dd, J= 7.6 Hz, J= 1.2 Hz, 1H), 7.58 (td, J = 7.6, = 1.2 Hz, 1H), 7.71 (td, J = 7.6, 7= 1.2 Hz, 1H), 7.91 (dd, J = 7.6, /= 1.2 Hz, 1H); MSiES ^") m/z 257 (MH⁺).

Step 6: To a solution of ethyl 2-(2-(ethyisulfonyl)phenyl)acetate (3.4 g) in ethanol (20 mL) was added a solution of NaOH (2.122 g) in water (20 mL). The reaction mixture was stirred at room temperature overnight. Ethanol was removed under reduced pressure, and 10 ml of water was added to the aqueous solution. The aqueous phase was washed with dichloromethane (30 mL x 3), and then acidified with 6 M HC1 to pH = 1. The solution was extracted with ethyl acetate (1000 mL x 3). The combined organic phase was washed with brine (50 mL), dried over sodium sulphate, filtered, and concentrated to give 2-(2-(ethylsulfonyl)phenyl)acetic acid (2 g) as a light yellow solid. 'H-NMR (400 MHz, DMSO</₆) δ ppm 1.10 (t, J = 7.2 Hz, 3H), 3.27 (q, J= 7,2 Hz, 2H), 4.06 (s, 2H), 7.51- 7.58 (m, 2H), 7.70 (td, J= 7.6, J= 1.2 Hz, 2H), 7.90 (dd, 7= 8.0, J= 1.2 Hz, 2H), 12.48 (s, 1 H); MS(ES^*) m/z 229 (MH⁺).

Preparation of the final product

A mixture of (2-amino-4-phenyl-l,3-thiazol-5-yl)(phenyl)methanone (intermediate la, 80 mg), [2-(ethylsulfonyl)phenyl]acetic acid (intermediate 11a, 68.4 mg), EDC (71.1 mg) and HOBt (50.1 mg) in dichloromethane (DCM) (3.5 mL) was stirred at room temperature overnight. Solvent was removed under reduced pressure and the residue was purified by MDAP to afford 2-[2-

(ethylsulfonyl)phenyl]-N-[4-phenyl-5-(phenylcarbonyl)-l ,3-thiazol-2-yl]acetamide (62 mg) as a white solid. ¹ H-NMR (400 MHz, DMSC δ ppm 1.13 (t, J= 7.3 Hz, 3H), 3.32 (q, J= 7.3 Hz, 2H), 4.36 (s, 2H), 7.26-7.17 (m, 5H), 7.44-7.38 (m, 3H), 7.63-7.53 (m, 4H), 7.74 (dt, J= 1.3 Hz, J- 7.5 Hz, 1H), 7.93 (dd, J= 1.2 Hz, /= 7.9 Hz, 1H), 12.97 (s, 1H); MS(ES⁺) m/z 491 ( H⁺).

Example 12

-JS-iethylsulfonylJphenylJ-A'- - henyl-S-iphenylcarbony^-l^-thiazol- -yljacetamide

Intermediate 12a: 2-faromo-l-(2,6-dichlorophenyl ethanone

Step 1 : A solution of sodium nitrite (0.916 g) in 20 mL of water was added dropwise to a suspension of 2-(3-aminophenyl)acetic acid (2 g) in 20 mL of water and 2.7 mL of concentrated hydrochloric acid cooled at 0 °C. After the addition was complete, the reaction mixture was stirred at the same temperature for a further 45 minutes. This cold diazonium salt solution was then added dropwise to a mixture of potassium O-ethyl carbonodithioate (2.456 g), 20 mL of water and 10 mL of a 2 M sodium carbonate solution at room temperature. The reaction mixture was heated at 45 °C until gas evolution stops. The mixture was subsequently cooled to room temperature, the pH was adjusted to 1 with concentrated hydrochloric acid. The oiled xanthogenate ester was extracted with diethyl ether, Solvent was evaporated to give a dark red liquid 2-(3-(ethoxycarbonothioylthio)phenyl)acetic acid (4.8 g). MS(ES⁺) m/z 257 (MH⁺).

Step 2: 2-(3-(ethoxycarbonothioylthio)phenyl) cetic acid (4.8 g) was taken up in 50 mL of ethanol, and a solution of OH (1.051 g) in 50 mL of water was added. The reaction mixture was heated at reflux for 20 hours. The major portion of ethanol was subsequently removed by distillation at reduced pressure. The aqueous phase was cooled with ice, and was rendered acid with

concentrated hydrochloric acid while stirring well. The solution was extracted with diethyl ether (100 mL). The organic phase was washed with brine, dried over anhydrous sodium sulphate, filtered, and concentrated to afford the desired product 2-(3-mercaptophenyl)acetic acid (3.3 g) as a brown solid. MS(ES⁺) m/z 169 (MH⁺).

Step 3: To a solution of 2-(3-mercaptophenyl)acetic acid (3.3 g) in N,N-dimethylformarnide (DMF) (50 mL) was added .₂C0₃ (10.85 g) and bromoethane (4.39 mL). The reaction mixture was stirred at room temperature for 2,5 hours. The reaction mixture was partitioned between ethyl acetate (100 mL) and water (100 mL). The organic was washed with water (100 mL x 4) and brine (200 mL), dried with sodium sulfate, filtered, and concentrated to give the desired product ethyl 2-(3-

(ethylthio)phenyl)acetate (2.76 g) as a pale yellow solid. Ή-NMR (400 MHz, DMSO-</₆) δ ppm 1.16 (t, J- 9.6 Hz, 3H), 1.23 (t, J = 9.6 Hz, 3H), 2.97 (q, J= 9.6 Hz, 2H), 3.65 (s, 2H), 4.08 (q, J= 9.6 Hz, 2H), 7.07 (d, J= 9.6 Hz, 1H), 7.1 -7.29 (m, 3H); MS(ES⁺) m/z 225 (MH⁺).

Step 4: Ethyl 2-(3-(ethylthio)phenyl)acetate (2.76 g) was dissolved in dichloromethane (DCM) (50 mL), and the solution was cooled to 0 °C with an ice bath. MCPBA (6.37 g) was added in portions, and the reaction mixture was stirred at room temperature overnight. The reaction mixture was filtered to removed the solid. The filtrate was washed with sat. sodium carbonate solution (50 mL x 2), water (50 mL), brine (150 mL), dried over sodium sulfate, filtered, and concentrated. The residue was purified by column chromatography (silica gel; EtOAc / PE = 1 :6) to afford the target compound ethyl 2-(3-(ethyIsulfonyI)phenyl)acetate (2.2 g) as a yellow oil. Ή-NMR (400 MHz,

DMSC ) δ ppm 1.10 (t, J= 7.2 Hz, 3H), 1.19 (t, J= 7.2 Hz, 3H), 3.28 (q, J= 7.2 Hz, 2H), 3.86 (s, 2H), 4.10 (q, J= 7.2 Hz, 2H), 7.60-7.67 (m, 2H), 7.78-7.82 (m, 2H); MS(ES⁺) m/z 257 (MH⁺).

Step 5: To a solution of ethyl 2-(3-(ethylsulfonyl)phenyl)acetate (2.2 g) in ethanol (20 mL) was added a solution of NaOH (1.373 g) in water (20 mL). The reaction mixture was stirred at room temperature overnight. Ethanol was removed under reduced pressure, and 10 ml of water was added to the aqueous solution. The aqueous phase was washed with dichloromethane (30 mL x 3), and then acidified with 6 M HC1 to pH = 1. The solution was extracted with ethyl acetate (50 mL x 3), The combined organic phase was washed with brine (50 mL), dried over sodium sulphate, filtered, and concentrated. The residue was purified by column chromatography (silica gel; MeOH : DCM = 1 :20) to give 2-(3-(ethylsulfonyl)phenyl)acetic acid (1 ,9 g) as a light yellow oil. Ή-NMR (400 MHz, DMSO-£ _fi) δ ppm 1.10 (t, J = 7.2 Hz, 3H), 3.29 (q, / = 7.2 Hz, 2H), 3.76 (s, 2H), 7.59-7.66 (m, 2H), 7.76-7.80 (m, 2H), 12.49 (s, 1H); MS(ES⁺) m/z 229 (MH⁺). Preparation of the final product

A mixture of (2-amino-4-phenyI-l,3-thiazol-5-yl)(phenyi)methanone (intermediate la, 80 mg), [3-(ethyIsulfonyi)phenyl]acetic acid (intermediate 12a, 71.7 mg), EDC (71.1 mg) and HOBt (50.1 mg) in dichloromethane (DCM) (3.5 mL) was stirred at room temperature overnight. Solvent was removed under reduced pressure and the residue was purified by MDAP to afford 2-[3-

(ethylsulfonyl)phenyl]-N-[4-phenyl-5-(phenylcarbonyl)-l,3-thiazol-2-yl]acetamide (80 mg) as a white solid. 'H-NMR (400 MHz, DMSO-c/₆) δ ppm 1.11 (t, J- 7.3 Hz, 3H), 3.30 (q, J= 7,3 Hz, 2H), 4.02 (s, 2H), 7.28-7.16 (m, 5H), 7.39-7.37 (m, 2H), 7.43 (t, J- 7.4 Hz, 1H), 7.56-7.54 (m, 2H), 7.65 (t, J= 7.7 Hz, 1 H), 7.73 (d, J= 7.8 Hz, 1H), 7.83-7.80 (m, 1H), 7.91 (s, 1H), 13.06 (s, 1H); MS(ES⁺) m/z 491 (MH⁺).

Biological Data

As stated above, the compounds according to Formula I are RORy modulators, and are useful in the treatment of diseases mediated by RORy. The biological activities of the compounds according to Formula I can be determined using any suitable assay for determining the activity of a candidate compound as a RORy modulator, as well as tissue and in vivo models.

Dual Fluorescence Energy Transfer (FRET) Assay

This assay is based on the knowledge that nuclear receptors interact with cofactors (transcription factors) in a ligand dependent manner. RORy is a typical nuclear receptor in that it has an AF2 domain in the ligand binding domain (LBD) which interacts with co-activators. The sites of interaction have been mapped to the LXXLL motifs in the co-activator SRC1(2) sequences. Short peptide sequences containing the LXXLL motif mimic the behavior of full-length co-activator.

The assay measures ligand-mediated interaction of the co-activator peptide with the purified bacterial-expressed RORy ligand binding domain (RORy-LBD) to indirectly assess ligand binding. RORy has a basal level of interaction with the co-activator SRC1 (2) in the absence of ligand, thus it is possible to find ligands that inhibit or enhance the RORy/SRCl (2) interaction.

Materials

Generation o/RORj-LBD bacterial expression plasmid

Human RORy Ligand Binding Domain (RORy-LBD) was expressed in E.coli strain

BL21(DE3) as an amino-terminal polyhistidine tagged fusion protein. DNA encoding this

recombinant protein was sub-cloned into a modified pET21a expression vector ( ovagen). A modified polyhistidine tag (MKKHHHHHHLVPRGS) was fused in frame to residues 263-518 of the human RORy sequence.

Protein Purification

Approximately 50 g E.coli cell pellet was resuspended in 300 ml of lysis buffer (30 ni imidazole pH 7.0 and 150 mM NaCl). Cells were lysed by sonication and cell debris was removed by centrifugation for 30 minutes at 20,000g at 4°C. The cleared supernatant was filtered through a 0.45 uM cellulose acetate membrane filter. The clarified lysate was loaded onto a column (X -26) packed with ProBond Nickel Chelating resin (InVitrogen), pre-equilibrated with 30 mM imidazole pH 7.0 and 150 mM NaCl. After washing to baseline absorbance with the equilibration buffer, the column was developed with a gradient from 30 to 500 mM imidazole pH 7.0. Column fractions containing the RORy-LBD protein were pooled and concentrated to a volume of 5 mis. The concentrated protein was loaded onto a Superdex 200 column pre-equilibrated with 20 mM Tris-CI pH 7.2 and 200 mM NaCl. The fractions containing the desired RORy-LBD protein were pooled together.

Protein Biotinylation

Purified RORy-LBD was buffer exchanged by exhaustive dialysis [3 changes of at least 20 volumes (>8000x)] against PBS [l OOmM NaPhosphate, pH 8 and 150mM NaCl]. The concentration of RORy-LBD was approximately 30uM in PBS. Five-fold molar excess of NHS-LC-Biotin (Pierce) was added in a minimal volume of PBS. This solution was incubated with occasional gentle mixing for 60 minutes at ambient room temperature. The modified RORy-LBD was dialyzed against 2 buffer changes - TBS pH 8.0 containing 5mM DTT, 2mM EDTA and 2% sucrose - each at least 20 times of the volume. The modified protein was distributed into aliquots, frozen on dry ice and stored at -80°C. The biotinylated RORy-LBD was subjected to mass spectrometric analysis to reveal the extent of modification by the biotinylation reagent. In general, approximately 95% of the protein had at least a single site of biotinylation and the overall extent of biotinylation followed a normal distribution of multiple sites ranged from one to five.

A biotinylated peptide corresponding to amino acid 676 to 700

(CPSSHSSLTERHKILHRLLQEGSPS) of the co-activator steroid receptor coactivator SRC1(2) was generated using similar method.

Assay

Preparation of Europium labeled SRC 1 (2) peptide: biotinylated SRC 1(2) solution was prepared by adding an appropriate amount of biotinylated SRC 1(2) from the lOOuM stock solution to a buffer containing 10 mM of freshly added DTT from solid to give a final concentration of 40 nM. An appropriate amount of Europium labeled Streptavidin was then added to the biotinylated SRC 1(2) solution in a tube to give a final concentration of 10 nM. The tube was inverted gently and incubated for 15 minutes at room temperature. Twenty-fold excess biotin from the 10 mM stock solution was added and the tube was inverted gently and incubated for 10 minutes at room temperature.

Preparation of APC labeled RORy-LBD: biotinylated RORy-LBD solution was prepared by adding an appropriate amount of biotinylated RORy-LBD from the stock solution to a buffer containing 10 mM of freshly added DTT from solid to give a final concentration of 40 nM. An appropriate amount of APC labeled Streptavidin was then added to the biotinylated RORy-LBD solution in a tube to give a final concentration of 20 nM. The tube was inverted gently and incubated for 15 minutes at room temperature. Twenty-fold excess biotin from the 10 mM stock solution was then added and the tube was inverted gently and incubated for 10 minutes at room temperature.

Equal volumes of the above-described Europium labeled SRC 1(2) peptide and the APC labeled RORy-LBD were gently mixed together to give 20nM RORy-LBD, ΙΟηΜ APC-Strepavidin, 20nM SRC 1(2) and 5nM Europium-Streptavidin. The reaction mixtures were incubated for 5 minutes. Using a Thermo Combi Multidrop 3S4 stacker unit, 25 u! of the reaction mixtures per well was added to the 384-well assay plates containing lul of test compound per well in 100% DMSO. The plates were incubated for lhour and then read on ViewLux in Lance mode for EU/APC.

Jurkat Cell Luciferase Assay

RORy is known to bind to a CNS (conserved non-coding sequences) enhancer element in the IL17 promoter. In this assay, RORy activity is indirectly assessed using a luciferase reporter construct which contains the human IL17 promoter having the RORy-specific CNS enhancer element. Inhibition of RORy activity by a compound will result in a decrease in luciferase activity of Jurkat cells transfected with the reporter construct.

Materials

Jurkat cell line

For the luciferase reporter plasmid, the 3 Kb human IL17 promoter containing the RORy- specific CNS enhancer element was PCR amplified from human genomic DNA and cloned into a pGL4-Luc2/hygro reporter plasmid sequencially as Xhol-Hindlll (1.1 Kb) and Kpnl-Xhol (1.9 Kb) fragments. For the 1.1 Kb fragment, PCR was used to amplify human IL17 proximal promoter region from genomic DNA of 293T cells using primers as follows: forward primer, 5 -

CTCGAGTAG AGCAGGAC AGGGAGGAA-3 ' (Xhol site is underlined) and reverse primer, 5'- AAGCTTGG ATG G ATG AGTTTGTGC CT-3 ' (Hind!II site is underlined). The 1.1 kb DNA bands were excised, purified, and inserted into pMD19-T Simple Vector (Takara). After DNA sequencing confirmation, the 1.1 kb DNA was digested with Xhol and Hindlll and inserted into XhoI/Hind!II sites of pGL4.31 [Iuc2P/GAL4UAS/Hygro] (Promega) to generate the plL17-lkb-luc reporter construct. For the 1.9 Kb fragment, PCR was used to amplify human IL17 promoter region from genomic DNA using primers as follows: forward primer 5 -

GGTACCTGCCCTGCTCTATCCTGAGT-3' (Kpnl site is underlined) and reverse primer, 5'- CTCGAGTGGTGAGTGCTGAGAGATGG-3 ' (Xhol site is underlined). The resulting 1.9 kb DNA bands were excised, gel purified, and cloned into a pMD1 -T Simple Vector (Takara). DNA sequencing analysis revealed that there were three point mutations but none of which affected RORyt binding. The 1.9 kb DNA fragment was released by double digestion with Kpnl and Xhol and inserted into pIL17-lkb-luc to generate the luciferase reporter plasmid "pIL17-3kb-CNS-luc." To overexpress RORyt, the full-length cDNA of human RORyt identical to the published sequence NM_001001523 was cloned into pcDNA3.1 at the Kpnl-Notl cloning sites to generate the RORyt overexpression plasmid "CDNA3.1DhRORy49-8".

The luciferase reporter plasmid and the RORyt overexpression plasmid were transfected into Jurkat cell line and a stable clone was identified. The stable clone was grown in 10% dialyzed FBS in RPMI (1640) with 800ug ml geneticin and 400ug/ml hygromecin.

Assay

Compounds were dissolved in DMSO at three concentrations, lOmM, 400uM and 16uM, and were dispensed into 384-wells assay plate at 40nl, 12.5nl, 5nl respectively. The volume was adjusted with pure DMSO to a give a final uniform volume of 40 nl Jurkat cells described above were counted and centrifuged. The growth medium was discarded and the cells were resuspended with assay medium (phenol red free RPMI) at lE-6/ml. Cells were added to each of the compounds in the assay plates. Cells were either untreated or treated with CD3 microbeads (Miltenyi Biotec) at 1 ul beads per 500,000 cells. Cells were culture overnight and luciferase assay (Promega) was performed. Data were collected by ViewLux (using luciferase greiner 384 setting).

Thl7 ELISA/Intracellular Staining Assays

ELISA

CD4+ cells were isolated from splenocytes of C57BL/6 (B6) mice (Shanghai Laboratory Animal Resource Center) using the CD4+ T Cell Isolation II Kit according to manufacturer's instructions (Miltenyi Biotec), 96 well plates were pre-coated with anti-CD3 antibody. CD4+ cells were resuspended in RPMI complete medium and were added to the 96-well plates at 3xl0⁵ cells/well, with the total volume of each well being 90 ul. A cytokine cocktail (90 ul) was then added to stimulate Thl7 differentiation. Each compound diluted to various concentrations in DMSO (DMSO final volume 0.1 %) was then added. The final concentrations of antibodies (R&D Systems) and cytokines (R&D Systems) were: anti-mCD3, 5ug/ml; anti-mCD28, 2ug ml; anti-mlFNy, lOug/ml; anti-mIL4, lOug/ml; mIL-6, 20ng ml; mIL-23, l Ong ml; mIL-Ι β, lOng/ml; TGF-β, lOng/ml. The cell culture was incubated at 37°C for 3 days. Supernatants were collected and IL-17 concentration was determined by ELISA, performed according to manufacturer's instructions (R&D Systems). The optical density (OD) at 405 nm were measured with a microplate reader (BioRad) and the IL-17 quantity were extrapolated from the standard curve. The percentage of IL-17 inhibition was calculated by referring to the positive control (100%) and the pIC50 were determined by GraphPad.

Intracellular staining

The Thl 7 differentiation cell culture described above was maintained for 5 days instead of 3 days. The effect of compounds on the production of IL-17 and IFN-γ in the ceils was determined by intracellular staining according to manufacturer's instructions (BD Biosciences).

As shown by ELISA and intracellular staining, the RORy modulator of Example 2 significantly reduced IL-17 production in Thl7 cells (Figure 1).

Assay Data

The data described below represents a mean pIC50 value of multiple test results if the test was perfonned more than once. It is understood that the data illustrated below may have reasonable variation depending on the specific conditions and procedures used by the person conducting the testing.

As shown in the Table below, all exemplified compounds (Examples 1-12) were tested in the dual FRET assay described above and were found to have a pIC50 between 6 and 9.

As shown in the Table below, all exemplified compounds (Examples 1-12) were tested in the Jurkat cell luciferase assay described above and were found to have a pIC50 between 5 and 7.

As shown in the Table below, all exemplified compounds except Examples 3, 11 and 12 were tested in the Thl 7 ELISA assays described above. All tested compounds except Example 7 were found to have a pIC50 between 5 and 7. Example 7 was tested twice and had a pIC50 below 5, the detection limit of the assay, in both tests. Example ROR Dual FRET Assay Jurkat Cell Luciferase Assay Thl7 Assay

No (pICSO) (pICSO) (pICSO)

1 7.9 6.5 5.5

2 8.1 6.9 6.6

3 6.7 5.8

4 7.6 6.5 5.5

5 8.0 6.9 6.5

6 7.6 6.6 6.0

7 8.1 6.9 <5

8 7.8 6.6 6.1

9 7.9 6.4 5.8

10 7.5 6.5 5.3

1 1 6.4 6.4

12 6.8 6.1

EAE Studies

Wild-type mice of the C57BL/6 (B6) strain were obtained from Shanghai Laboratory Animal Resource Center. EAE was induced by intravenous injections of 100 ng of pertussis toxin (List Biological Laboratories) and then subcutaneous immunization with 200 μΐ of an emulsion composed of MOG35.55 peptide (300 £/ΐΒ0ϋ8ε) in PBS and an equal volume of complete Freund's adjuvant containing 5 mg ml heat-killed Mycobacterium tuberculosis H37Ra (Difco Laboratories) on day 0, followed by another intravenous injections of 100 ng of pertussis toxin on day 2 as described previously (Wang et al. (2006) J. Clin. Invest. 116: 2434-2441). Each compound was given orally on day 0 at 100 mg/kg twice a day. Mice were scored for disease severity daily using a EAE scoring system (Wang et al. (2006) J. Clin. Invest. 116: 2434-2441): 0, no overt signs of disease; 1 , limp tail or hind limb weakness but not both; 2, limptail and paraparesis (weakness, incomplete paralysis of one or two hind limbs); 3, paraplegia (complete paralysis of two hind limbs); 4, paraplegia with forelimb weakness or paralysis; and 5, moribund state or death. Clinical score data were expressed as means ± s.e.m.

As shown in Figure 2, the RORy modulator of Example 2 delayed EAE onset,

CIA Studies

Collagen-induced arthritis (CIA) was induced in 8-week old male DBA/1 mice via an initial intradermal (i.d.) injection of an emulsion consisting of bovine type II collagen in CFA. Mice were intraperitoneally (i.p.) injected with bovine type II collagen 21 days later to boost the immune system, resulting in chronic inflammation in both the hind and the front paws. Each compound was given to the mice at lOOmg/kg twice a day starting from day 20 after the first immunization. Mice were examined for onset and severity of disease in a blinded manner. Arthritis symptoms were graded by the following scoring system: grade 0, normal appearance; grade 1 , slight erythema/ edema (1-3 digits); grade 2, erythema/ edema in more than 3 digits or mild swelling in ankle/wrist joint; grade 3, erythema/ edema in entire paw; grade 4, massive erythema/ edema of entire paw extending into proximal joints, ankylosis, loss of function. Each limb was graded, giving a maximum possible score of 16 per mouse. Clinical score data were expressed as means ± s.e.m. Foot volume of the mice was determined using a YLS-7B foot volume measuring instrument (Shandong Academy of Medical Science).

Methods of Use

The compounds of the invention are modulators of RORy and can be useful in the treatment of diseases mediated by RORy, particularly autoimmune or inflammatory diseases. The Inflammatory or autoimmune diseases of the invention include multiple sclerosis, rheumatoid arthritis, psoriasis, Crohn's disease, inflammatory bowel disease, Sjorgen's syndrome, optic neuritis, chronic obstructive pulmonary disease and type I diabetes, neuromyelitis optica, Myasthenia Gavis, uveitis, Guillain- Barre syndrome, psoriatic arthritis, Gaves' disease, asthma, chronic obstructive pulmonary disease and allergy. Accordingly, in another aspect the invention is directed to methods of treating such diseases.

The methods of treatment of the invention comprise administering a safe and effective amount of a compound according to Formula I or a pharmaceutically-acceptable salt thereof to a patient in need thereof.

As used herein, "treat" in reference to a condition means: (1) to ameliorate or prevent the condition or one or more of the biological manifestations of the condition, (2) to interfere with (a) one or more points in the biological cascade that leads to or is responsible for the condition or (b) one or more of the biological manifestations of the condition, (3) to alleviate one or more of the symptoms or effects associated with the condition, or (4) to slow the progression of the condition or one or more of the biological manifestations of the condition.

As indicated above, "treatment" of a condition includes prevention of the condition. The skilled artisan will appreciate that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially diminish the likelihood or severity of a condition or biological manifestation thereof, or to delay the onset of such condition or biological manifestation thereof.

As used herein, "safe and effective amount" in reference to a compound of the invention or other pharmaceutically-active agent means an amount of the compound sufficient to treat the patient's condition but low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment. A safe and effective amount of a compound will vary with the particular compound chosen (e.g. consider the potency, efficacy, and half-life of the compound); the route of administration chosen; the condition being treated; the severity of the condition being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient to be treated; the duration of the treatment; the nature of concurrent therapy; the desired therapeutic effect; and like factors, but can nevertheless be routinely determined by the skilled artisan.

As used herein, "patient" refers to a human or other animal.

The compounds of the invention may be administered by any suitable route of administration, including both systemic administration and topical administration. Systemic administration includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation. Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion. Inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages. Topical administration includes application to the skin as well as intraocular, otic, intravaginal, and intranasal administration.

The compounds of the invention may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or indefinitely to maintain the desired therapeutic effect. Suitable dosing regimens for a compound of the invention depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half-life, which can be determined by the skilled artisan. In addition, suitable dosing regimens, including the duration such regimens are administered, for a compound of the invention depend on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change.

Typical daily dosages may vary depending upon the particular route of administration chosen. Typical daily dosages for oral administration range from 0.1 mg to 1000 mg.

Additionally, the compounds of the invention may be administered as prodrugs. As used herein, a "prodrug" of a compound of the invention is a functional derivative of the compound which, upon administration to a patient, eventually liberates the compound of the invention in vivo. Administration of a compound of the invention as a prodrug may enable the skilled artisan to do one or more of the following: (a) modify the onset of the compound in vivo; (b) modify the duration of action of the compound in vivo; (c) modify the transportation or distribution of the compound in vivo; (d) modify the solubility of the compound in vivo; and (e) overcome or overcome a side effect or other difficulty encountered with the compound. Typical functional derivatives used to prepare prodrugs include modifications of the compound that are chemically or enzymatically cleaved in vivo. Such modifications, which include the preparation of phosphates, amides, esters, thioesters, carbonates, and carbamates, are well known to those skilled in the art.

In one embodiment, the invention relates to the use of the compounds of the invention in the preparation of a medicament for the treatment of diseases mediated by RORy. In another embodiment, the invention relates to the compounds of the invention for use in the treatment of diseases mediated by RORy. Examples of such diseases include autoimmune or inflammatory diseases such as multiple sclerosis, rheumatoid arthritis, psoriasis, Crohn's disease, inflammatory bowel disease, Sjorgen's syndrome, optic neuritis, chronic obstructive pulmonary disease and type I diabetes, neuromyelitis optica, Myasthenia Gavis, uveitis, Guillain-Barre syndrome, psoriatic arthritis, Gaves' disease, asthma, chronic obstructive pulmonary disease and allergy.

Compositions

The compounds of the invention will normally, but not necessarily, be formulated into pharmaceutical compositions prior to administration to a patient. Accordingly, in another aspect the invention is directed to pharmaceutical compositions comprising a compound of the invention and one or more pharmaceutically-acceptable excipient.

The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein a safe and effective amount of a compound of the invention can be extracted and then given to the patient such as with powders or syrups. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form wherein each physically discrete unit contains a safe and effective amount of a compound of the invention. When prepared in unit dosage form, the pharmaceutical compositions of the invention typically contain from 0.1 mg to 1000 mg.

The pharmaceutical compositions of the invention typically contain one compound of the invention. However, in certain embodiments, the pharmaceutical compositions of the invention contain more than one compound of the invention. For example, in certain embodiments the pharmaceutical compositions of the invention contain two compounds of the invention. In addition, the pharmaceutical compositions of the invention may optionally further comprise one or more additional pharmaceutically active compounds.

As used herein, "pharmaceutically-acceptable excipient'¹ means a pharmaceutically acceptable material, composition or vehicle involved in giving form or consistency to the pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when commingled such that interactions which would substantially reduce the efficacy of the compound of the invention when administered to a patient and interactions which would result in pharmaceutical compositions that are not pharmaceutically acceptable are avoided. In addition, each excipient must of course be of sufficiently high purity to render it pharmaceutically-acceptable.

The compound of the invention and the pharmaceutically-acceptable excipient or excipients will typically be formulated into a dosage form adapted for administration to the patient by the desired route of administration. For example, dosage forms include those adapted for (1) oral administration such as tablets, capsules, caplets, pills, troches, powders, syrups, elixers, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation such as dry powders, aerosols, suspensions, and solutions; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.

Suitable pharmaceutically-acceptable excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceutically-acceptable excipients may be chosen for a particular function that they may serve in the composition. For example, certain pharmaceutically- acceptable excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutically-acceptable excipients may be chosen for their ability to facilitate the carrying or transporting of the compound or compounds of the invention once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body. Certain pharmaceutically-acceptable excipients may be chosen for their ability to enhance patient compliance.

Suitable pharmaceutically-acceptable excipients include the following types of excipients: Diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emulsifiers, sweetners, flavoring agents, flavor masking agents, coloring agents, anticaking agents, hemectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. The skilled artisan will appreciate that certain pharmaceutically-acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.

Skilled artisans possess the knowledge and skill in the art to enable them to select suitable pharmaceutically-acceptable excipients in appropriate amounts for use in the invention. In addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically-acceptable excipients and may be useful in selecting suitable pharmaceutically- acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The pharmaceutical compositions of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

In one aspect, the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of the invention and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystalline cellulose), calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g. corn starch, potato starch, and pre-gelatinized starch), gelatin, acacia, sodium alginate, alginic acid, tragacanth, guar gum, povidone, and cellulose and its derivatives (e.g. microcrystalline cellulose). The oral solid dosage fonn may further comprise a disintegrant. Suitable disintegrants include crospovidone, sodium starch glycolate, croscarmelose, alginic acid, and sodium carboxyraethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, magnesuim stearate, calcium stearate, and talc.

Claims

AT IS CLAIMED IS:

A method of treatment of diseases mediated by RORy which comprises administering to a host in need thereof an effective amount of a compound according to Formula I or a pharmaceutically acceptable salt thereof

Formula I

wherein:

Rl is H or C l -C6 alkyi;

R2 is H or Cl -C6 alkyl; and

R3 is selected from the group consisting of:

- C1-C6 alkyl optionally substituted with OH, C1-C3 alkoxy or one to three F; and

- NHRa wherein Ra is H or C1 -C3 alkyl.

2. A method according to claim 1 , wherein Rl and R2 are each H.

3. A method according to claim 1 , wherein one of Rl and R2 is methyl.

4. A method according to claim 1 , wherein both Rl and R2 are methyl.

5. A method according to any of claims 1 to 4, wherein R3 is C1-C3 alkyl optionally substituted with OH, C1-C3 alkoxy or CF₃.

6. A method according to any of claims 1 to 4, wherein R3 is methylamino or ethylamino.

7. A method according to any of claims 1 to 6, wherein said disease is multiple sclerosis.

8. A method according to any of claims 1 to 6, wherein said disease is rheumatoid arthritis.

9. A compound of Formula 1(a) or a pharmaceutically acceptable salt thereof

Formula 1(a)

wherein:

Rl is H; R2 is H; and

R3 is selected from the group consisting of:

- C1 -C6 alky] substituted with OH, C1-C3 alkoxy or one to three F; and

- NHRa wherein Ra is H or C1-C3 alkyl.

or

Rl is H or C1-C3 alkyl, R2 is H or C1-C3 alkyl, wherein at least one of Rl and R2 is not H; and

R3 is C1-C6 alkyl.

10. A compound or salt according to claim 9, wherein Rl is H, R2 is H, and R3 is methylamino or ethylamino.

1 1. A compound or salt according to claim 9, wherein Rl is H, R2 is H, and R3 is CI -C3 alkyl substituted with OH, methoxy or CF₃.

12. A compound or salt according to claim 9, wherein at least one of Rl and R2 is methyl.

13. A pharmaceutical composition which comprises a compound according to any of claims 9 to 12 and a pharmaceutically acceptable carrier or excipient.

14. A compound according to any of claims 9 to 12 for use in the treatment of multiple sclerosis.

15. A compound according to any of claims 9 to 12 for use in the treatment of rheumatoid arthritis.

16. A pharmaceutical composition for use in the treatment of multiple sclerosis which comprises a compound according to any one of claims 9 to 12 and a pharmaceutically acceptable carrier.

17. A pharmaceutical composition for use in the treatment of rheumatoid arthritis which comprises a compound according to any one of claims 9 to 12 and a pharmaceutically acceptable carrier.