Si.
Novel Thiophene Derivatives
Field of the invention
The present invention relates to S1P1/EDG1 receptor agonists of Formula (I) and their use as active ingredients in the preparation of pharmaceutical compositions. The invention also concerns related aspects including processes for the preparation of the compounds, pharmaceutical compositions containing a compound of the Formula (I), and their use as compounds improving vascular function and as immunomodulating agents, either alone or in combination with other active compounds or therapies. A further aspect of the invention relates to novel compounds of Formulae (II) and (III) that serve as intermediates to prepare compounds of Formula (I).
Background of the invention
The human immune system is designed to defend the body against foreign micro¬organisms and substances that cause infection or disease. Complex regulatory mechanisms ensure that the immune response is targeted against the intruding substance or organism and not against the host. In some cases, these control mechanisms are unregulated and autoimmune responses can develop. A consequence of the uncontrolled inflammatory response is severe organ, cell, tissue or joint damage. With current treatment, the whole immune system is usually suppressed and the body's ability to react to infections is also severely compromised. Typical drugs in this class include azathioprine, chlorambucil, cyclophosphamide, cyclosporin, or methotrexate. Corticosteroids which reduce inflammation and suppress the immune response, may cause side effects when used in long term treatment. Nonsteroidal anti-infammatory drugs (NSAIDs) can reduce pain and inflammation, however, they exhibit considerable side effects. Alternative treatments include agents that activate or block cytokine signaling.
Orally active compounds with immunomodulating properties, without compromising immune responses and with reduced side effects would significantly improve current treatments of uncontrolled inflammatory disease.

In the field of organ transplantation the host immune response must be suppressed to prevent organ rejection. Organ transplant recipients can experience some rejection even when they are taking immunosuppressive drugs. Rejection occurs most frequently in the first few weeks after transplantation, but rejection episodes can also happen months or even years after transplantation. Combinations of up to three or four medications are commonly used to give maximum protection against rejection while minimizing side effects. Current standard drugs used to treat the rejection of transplanted organs interfere with discrete intracellular pathways in the activation of T-type or B-type white blood cells. Examples of such drugs are cyclosporin, daclizumab, basiliximab, everolimus, or FK506, which interfere with cytokine release or signaling; azathioprine or leflunomide, which inhibit nucleotide synthesis; or 15-deoxyspergualin, an inhibitor of leukocyte differentiation.
The beneficial effects of broad immunosuppressive therapies relate to their effects; however, the generalized immunosuppression which these drugs produce diminishes the immune system's defense against infecfion and malignancies. Furthermore, standard immunosuppressive drugs are often used at high dosages and can cause or accelerate organ damage.
Description of the invention
The present invention provides novel compounds of Formula (I) that are agonists for the G protein-coupled receptor S1P1/EDG1 and have a powerful and long-lasting immunosuppressive effect which is achieved by reducing the number of circulating and infiltrating T- and B-lymphocytes, without affecting their maturation, memory, or expansion. The reduction of circulating T- / B-lymphocytes as a result of S1P1/EDG1 agonism, possibly in combination with the observed improvement of endothelial cell layer function associated with S1P1/EDG1 activation, makes such compounds useful to treat uncontrolled inflammatory disease and to improve vascular functionality.
The compounds of the present invention can be utilized alone or in combination with standard drugs inhibifing T-cell acfivation, to provide a new immunosuppressive therapy with a reduced propensity for infections when

compared to standard immunosuppressive therapy. Furthermore, the compounds of the present Invention can be used in combination with reduced dosages of traditional Immunosuppressant therapies, to provide on the one hand effective immunosuppressive activity, while on the other hand reducing end organ damage associated with higher doses of standard immunosuppressive drugs. The observation of improved endothelial cell layer function associated with S1P1/EDG1 activation provides additional benefits of compounds to improve vascular function.
The nucleotide sequence and the amino acid sequence for the human S1P1/EDG1 receptor are known in the art and are published in e.g.: HIa, T., and Maciag, T. J. Biol Chem. 265 (1990), 9308-9313; WO 91/15583 published 17 October 1991; WO 99/46277 published 16 September 1999. The potency and efficacy of the compounds of Formula (I) are assessed using a GTP7S assay to determine EG50 values and by measuring the circulating lymphocytes in the rat after oral administration, respectively (see in Examples).
The general terms used hereinbefore and hereinafter preferably have, within this disclosure, the following meanings, unless otherwise indicated:
Where the plural form is used for compounds, salts, pharmaceutical compositions, diseases and the like, this is intended to mean also a single compound, salt, or the like.
Any reference hereinbefore or hereinafter to a compound of Formula (I), (II) or (III) is to be understood as referring also to salts, especially pharmaceutically acceptable salts, of a compound of Formula (I), (II) or (III), as appropriate and expedient.
The term Ci^-alkyl, alone or in combination with other groups, means saturated, branched or preferably straight chain groups with one to four carbon atoms, preferably one to three carbon atoms. Examples of Ci.4-alkyl groups are methyl, ethyl, /7-propyl, /so-propyl, /?-butyl, /so-butyl, sec-butyl, and te/Y-butyl.

The term Ci^-alkoxy, alone or in combination with other groups, means an R-0-group, wherein R is a Ci^-alkyl. Preferred examples of Ci.4-alkoxy groups are methoxy, ethoxy, propoxy, iso-propoxy, /so-butoxy, sec-butoxy and terf-butoxy.
The term hydroxy-C2.4-alkoxy means a straight or branched alkoxy chain bearing a hydroxy group whereby there are at least two carbon atoms between the hydroxy group and the oxygen of the C2^-alkoxy group. Examples of hydroxy-C2-4-alkoxy groups are 2-hydroxy-ethoxy, 3-hydroxy-propoxy, 2-hydroxy-propoxy, 4-hydroxy-butoxy, 3-hydroxy-1-methyl-propoxy, and 3-hydroxy-butoxy.
The term Ci^-alkylamino or di-(Ci^-alkyl)amino, alone or in combination with other groups, means an R'-NH- or an R'-NR"- group, wherein R' and R" are each independently a Ci-4-alkyl group. Preferred examples of Ci^-alkylamino or di-(Ci-4-alkyl)amino groups are methylamino, ethylamino, N,N-dimethylamino, and N-methyl-N-ethyl-amino.
The term halogen means fluoro, chloro, bromo or iodo, preferably fluoro or chloro.
Salts are preferably the pharmaceutically acceptable salts of the compounds of Formula (I).
Salt-forming groups are groups or radicals having basic or acidic properties. Compounds having at least one basic group or at least one basic radical, for ex¬ample amino, a secondary amino group not forming a peptide bond or a pyridyl radical, may form acid addition salts, for example with inorganic acids. When several basic groups are present mono- or poly-acid addition salts may be formed.
Compounds having acidic groups, such as a carboxy group or a phenolic hydroxy group, may form metal or ammonium salts, such as alkali metal or alkaline earth metal salts, for example sodium, potassium, magnesium or calcium salts, or ammonium salts with ammonia or suitable organic amines, such as tertiary monoamines, for example triethylamine or tri-(2-hydroxyethyl)-amine, or

heterocyclic bases, for example A/-ethyl-piperidine or A/,/\/'-dimethylplperazine. Mixtures of salts are possible.
Compounds having both acidic and basic groups can form internal salts.
For the purposes of isolation or purification, as well as in the case of compounds that are used further as intermediates, it is also possible to use pharmaceutically unacceptable salts, e.g. the picrates. Only pharmaceutically acceptable, non-toxic salts may be used for therapeutic purposes, however, and those salts are therefore preferred.
The expression pharmaceutically acceptable salts encompasses either salts with inorganic acids or organic acids like hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid, nitric acid, phosphorous acid, nitrous acid, citric acid, formic acid, acetic acid, oxalic acid, maleic acid, lactic acid, tartaric acid, fumaric acid, benzoic acid, mandelic acid, cinnamic acid, palmoic acid, stearic acid, glutamic acid, aspartic acid, methanesulfonic acid, ethanesulfonic acid, ethanedisulfonic acid, p-toluenesulfonic acid, salicylic acid, succinic acid, trifluoroacetic acid, and the like that are non toxic to living organisms or, in case the compound of Formula (I), (II) or (III) is acidic in nature, with an inorganic base like an alkali or earth alkali base, e.g. sodium hydroxide, potassium hydroxide, calcium hydroxide and the like. For other examples of pharmaceutically acceptable salts, reference can be made to "Salt selection for basic drugs". Int. J. Pharm. (1986), 33, 201-217.
The compounds of Formula (I) and (III) may contain one or more stereogenic or asymmetric centers, such as one or more asymmetric carbon atoms. Substituents at a double bond or a ring may be present in cis- or trans-form (E- or Z-form) unless indicated othenwise. The compounds of Formula (I) and (III) may thus be present as mixtures of stereoisomers or preferably as pure stereoisomers. Mixtures of stereoisomers may be separated in a manner known to a person skilled in the art.
i) The invention relates to novel thiophene compounds of the Formula (I),

dihydroxypropoxy, 2-hydroxy-3-methoxy-propoxy, -OCH2-(CH2)m-NR^^R^^ -OCH2-(CH2)m-NHC0R^^ -OCH2-(CH2)m-NHS02R^^ -0CH2-CH(0H)-CH2-NR''R^2 -OCH2-CH(0H)-CH2-NHC0R^, -OCH2-CH(OH)-CH2-NHS02R^^ -NR^^R^^ -NHCO-R^. or -S02NH-R^^
R®^ represents hydrogen, methyl, ethyl, 2-hydroxyethyl, 2-hydroxy-1-
hydroxymethyl-ethyl, 2,3-dihydroxy-propyi, 2-Ci-4-alkoxyethyl, 3-hydroxypropyl, 3-
Ci^-alkoxypropyl, 2-aminoethyl, 2-(Ci-(-alkylamino)ethyl, 2-(dl-(Ci.4-
alkyi)amino)ethyl, carboxymethyl, Ci-t-alkylcarboxymethyl, 2-carboxyethyl, or 2-(Ci. 4-alkylcarboxy)ethyl;
R" represents hydrogen, or methyl;
R" represents methyl, ethyl, 2-hydroxyethyl, 2-methoxyethyl, methylamino, ethylamino, or dimethylamino;
R^^ represents hydroxymethyl, aminomethyl, methylaminomethyl, dimethylamlnomethyl, 2-aminoethyr, or2-methylamlno-ethyl;
R^® represents hydrogen;
R®® represents hydrogen or hydroxy; and
in case R*^ represents hydroxy, R" may in addition represent hydroxy;
m represents the integer 1 or 2;
n represents 0, 1, or 2;
k represents 0;
p represents 0 or 1; and
in case p represents 1, k may in addition represent 1; and
R^ represents hydrogen, Ci^-alkyl, or halogen;
and salts thereof.

vii) Another particular embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to vi), wherein R"* represents hydrogen or methyl.
viii) Another particular embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to vi), wherein R^ represents hydrogen.
ix) Another particular embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to vi), wherein R"* represents a methyl group.
x) A preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to ix), wherein R^ represents an isobutyl group.
xi) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to x), wherein R^ represents methyl, ethyl, n-propyl, or isobutyl.
xii) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to x), wherein R^ represents a methyl group.
xiii) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xii), wherein R* represents methoxy, and R^ and R^ represent hydrogen, or wherein R* represents hydrogen and R* represents methyl, ethyl, or methoxy and R^ represents methyl, ethyl or halogen.

xiv) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xii), wherein R* represents a methoxy group, and R® and R' represent hydrogen.
XV) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xii), wherein R* represents hydrogen, and R® and R^ represent a methyl group.
xvi) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xii), wherein R* represents hydrogen, and R* and R^ represent an ethyl group.
xvii) A particularly preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xii), wherein R* represents hydrogen, R* represents a methyl group, and R' represents an ethyl group.
xviii) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xii), wherein R* represents hydrogen, R* represents a methoxy group, and R^ represents a chlorine atom.
xix) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xii), wherein R* represents hydrogen, R* represents a methyl group, and R^ represents a chlorine atom.
xx) Another preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xix), wherein R^ represents -(CH2)k-(CHR^^)p-CHR^^-CONR^^R^^
xxi) A particularly preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xix), wherein R®

represents -(CH2)k-(CHR^^)p-CHR^^-C0NR^^R^^ wherein k represents 0, ^ represents 1, and R®* and R®* represent hydrogen.
xxii) A preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments 1) to xix), wherein R^ represents -OCH2-CH(0H)-CH2-NHC0R^.
xxiii) A particularly preferred embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) to xix), wherein R® represents -OCH2-CH(OH)-CH2-NHCOR^'*, wherein R^* represents hydroxymethyl.
xxiv) A particular embodiment of the invention relates to thiophene derivatives according to any one of the embodiments i) and iii) to xix), wherein R« represents 2,3-dihydroxypropoxy.
xxv) Another particular embodiment of the invention relates to thiophene derivatives according to any one of the embodiments ii) to xix), wherein R^ represents 1-glyceryl.

2-hydroxy-N-(3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamicle;
3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-ylH1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propane-1,2-diol;
3-{4-[5-(3-ethyl-4-isobutyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propane-1,2-diol;
3-{4-[5-(4-isobutyl-3-propyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propane-1,2-diol;
3-{4-[5-(3,4-diisobutyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propane-1,2-diol;
2-hydroxy-N-(2-hydroxy-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamlde;
N-(3-{4-[5-(3-ethyl-4-isobutyl-thiophen-2-yl)-[1,3,4]oxadlazol-2-yl]-2,6-dimethyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-aGetamide;
2-hydroxy-N-(2-hydroxy-3-{4-[5-(4-isobutyl-3-propyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
N-(3-{4-[5-(3,4-dilsobutyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-3-methoxy-phenoxy}-propane-1,2-diol;
2-hydroxy-N-(2-hydroxy-3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-3-methoxy-phenoxy}-propyl)-acetamide; and
2-hydroxy-N-(2-hydroxy-3-{4-[5-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide.
xxx) Further especially preferred thiophene compounds according to Formula (I) are:
N-((S)-3-{2,6-diethyl-4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((S)-3-{2-chloro-4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((R)-3-{2-chloro-4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;

3-{2-ethyl-4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenyl}-N-(2-hydroxy-ethyl)-propionamide;
2-hydroxy-N-((S)-2-hydroxy-3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
2-hydroxy-N-((R)-2-hydroxy-3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
N-((R)-3-{2-ethyl-4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((S)-3-{2-ethyl-4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-(2-hydroxy-1-hydroxymethyl-ethyl)-3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenyl}-propionamide;
2-hydroxy-N-((S)-2-hydroxy-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
2-hydroxy-N-((R)-2-hydroxy-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
N-((S)-3-{2-ethyl-4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((R)-3-{2-ethyl-4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-({S)-3-{2-chloro-4-I5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-nnethyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((R)-3-{2-chloro-4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((S)-3-{2,6-diethyl-4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((R)-3-{2,6-diethyl-4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((S)-3-{2-ethyl-4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((R)-3-{2-ethyl-4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;

2-hyclroxy-N-((S)-2-hyclroxy-3-{4-[3-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxad iazol-5-yl]-2,6-d imethyl-phenoxy}-propyl)-aceta m ide;
2-hydroxy-N-((R)-2-hydroxy-3-{4-[3-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-5-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
2-hydroxy-N-((S)-2-hydroxy-3-{4-[3-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-3-oxo-propyl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
2-hydroxy-N-((R)-2-hydroxy-3-{4-[3-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-3-oxo-propyl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
N-((S)-3-{2-ethyl-4-[3-{4-isobutyl-3,5-dimethyl-thiophen-2-yl)-3-oxo-propyl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((R)-3-{2-ethyl-4-[3-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-3-oxo-propyl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((S)-3-{2-chloro-4-[5-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
N-((R)-3-{2-chloro-4-[5-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
2-hydroxy-N-((S)-2-hydroxy-3-{4-[5-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
2-hydroxy-N-((R)-2-hydroxy-3-{4-[5-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propyl)-acetamide;
N-((R)-3-{2-ethyl-4-[5-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide;
and
N-((S)-3-{2-ethyl-4-[5-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-6-methyl-phenoxy}-2-hydroxy-propyl)-2-hydroxy-acetamide.

20th Edition, Philadelphia College of Pharmacy and Science) by bringing the described compounds of Formula (I) or their pharmaceutically acceptable salts, optionally in combination with other therapeutically valuable substances, into a galenical administration form together with suitable, non-toxic, inert, pharmaceutically acceptable solid or liquid carrier materials and, if desired, usual pharmaceutical adjuvants.
The pharmaceutical compositions comprising a compound of Formula (I) are useful for the prevention and/or treatment of diseases or disorders associated with an activated immune system.
Such diseases or disorders are selected from the group consisting of rejection of transplanted organs, tissue or cells; graft-versus-host diseases brought about by transplantation; autoimmune syndromes including rheumatoid arthritis; systemic lupus erythematosus; antiphospholipid syndrome; Hashimoto's thyroiditis; lymphocytic thyroiditis; multiple sclerosis; myasthenia gravis; type I diabetes; uveitis; episcleritis; scleritis; Kawasaki's disease, uveo-retinitis; posterior uveitis; uveitis associated with Behcet's disease; uveomeningitis syndrome; allergic encephalomyelitis; chronic allograft vasculopathy; post-infectious autoimmune diseases including rheumatic fever and post-infectious glomerulonephritis; inflammatory and hyperproliferative skin diseases; psoriasis; psoriatic arthritis; atopic dermatitis; myopathy; myositis; osteomyelitis; contact dermatitis; eczematous dermatitis; seborrhoeic dermatitis; lichen planus; pemphigus; bullous pemphigoid; epidermolysis bullosa; urticaria; angioedema; vasculitis; erythema; cutaneous eosinophilia; acne; scleroderma; alopecia areata; keratoconjunctivitis; vernal conjunctivitis; keratitis; herpetic keratitis; dystrophia epithelialis corneae; corneal leukoma; ocular pemphigus; Mooren's ulcer; ulcerative keratitis; scleritis; Graves' ophthalmopathy; Vogt-Koyanagi-Harada syndrome; sarcoidosis; pollen allergies; reversible obstructive airway disease; bronchial asthma; allergic asthma; intrinsic asthma; extrinsic asthma; dust asthma; chronic or inveterate asthma; late asthma and airway hyper-responsiveness; bronchiolitis; bronchitis; endometriosis; orchitis; gastric ulcers; ischemic bowel diseases; inflammatory bowel diseases; necrotizing enterocolitis; intestinal lesions associated with thermal burns; coeliac disease;

proctitis; eosinophilic gastroenteritis; mastocytosis; Crohn's disease; ulcerative colitis; vascular damage caused by ischemic diseases and thrombosis; atherosclerosis; fatty heart; myocarditis; cardiac infarction; aortitis syndrome; cachexia due to viral disease; vascular thrombosis; migraine; rhinitis; eczema; Interstitial nephritis; IgA-induced nephropathy; Goodpasture's syndrome; hemolytic-uremic syndrome; diabetic nephropathy; glomerulosclerosis; glomerulonephritis; tubulointerstitlal nephritis; Interstitial cystitis; multiple myositis; Guillain-Barr6 syndrome; Meniere's disease; polyneuritis; multiple neuritis; myelitis; mononeuritis; radiculopathy; hyperthyroidism; Basedow's disease; thyrotoxicosis; pure red cell aplasia; aplastic anemia; hypoplastic anemia; idiopathic thrombocytopenic purpura; autoimmune hemolytic anemia; autoimmune thrombocytopenia; agranulocytosis; pernicious anemia; megaloblastic anemia; anerythroplasia; osteoporosis; fibroid lung; idiopathic interstitial pneumonia; dermatomyositis; leukoderma vulgaris; ichthyosis vulgaris; photoallergic sensitivity; cutaneous T cell lymphoma; polyarteritis nodosa; Huntington's chorea; Sydenham's chorea; myocardosis; myocarditis; scleroderma; Wegener's granuloma; Sjogren's syndrome; adiposis; eosinophilic fascitis; lesions of gingiva, periodontium, alveolar bone, substantia ossea dentis; male pattern alopecia or alopecia senilis; muscular dystrophy; pyoderma; Sezary's syndrome; hypophysltis; chronic adrenal insufficiency; Addison's disease; ischemla-reperfusion injury of organs which occurs upon preservation; endotoxin shock; pseudomembranous colitis; colitis caused by drug or radiation; ischemic acute renal insufficiency; chronic renal insufficiency; lung cancer; malignancy of lymphoid origin; acute or chronic lymphocytic leukemlas; lymphoma; pulmonary emphysema; cataracta; siderosis; retinitis pigmentosa; senile macular degeneration; vitreal scarring; corneal alkali burn; dermatitis erythema; ballous dermatitis; cement dermatitis; gingivitis; periodontitis; sepsis; pancreatitis; peripheral artery disease; carcinogenesis; solid cancer tumors; metastasis of carcinoma; hypobaropathy; autoimmune hepatitis; primary biliary cirrhosis; sclerosing cholangitis; partial liver resection; acute liver necrosis; cirrhosis; alcoholic cirrhosis; hepatic failure; fulminant hepatic failure; late-onset hepatic failure; and "acute-on-chronIc" liver failure.

Preferred diseases or disorders to be treated and/or prevented with the compounds of Formula (I) are selected from the group consisting of rejection of transplanted organs such as kidney, liver, heart, lung, pancreas, cornea, and skin; graft-versus-host diseases brought about by stem cell transplantation; autoimmune syndromes including rheumatoid arthritis, multiple sclerosis, inflammatory bowel diseases such as Crohn's disease and ulcerative colitis, psoriasis, psoriatic arthritis, thyroiditis such as Hashimoto's thyroiditis, uveo-retinitis; atopic diseases such as rhinitis, conjunctivitis, dermatitis; asthma; type I diabetes; post-infectious autoimmune diseases including rheumatic fever and post-infectious glomerulonephritis; solid cancers and tumor metastasis.
Particularly preferred diseases or disorders to be treated and/or prevented with the compounds of Formula (I) are selected from the group consisting of rejection of transplanted organs selected from kidney, liver, heart and lung; graft-versus-host diseases brought about by stem cell transplantation; autoimmune syndromes selected from rheumatoid arthritis, multiple sclerosis, psoriasis, psoriatic arthritis, Crohn's disease, and Hashimoto's thyroiditis; and atopic dermatitis.
The present invention also relates to a method for the prevention or treatment of a disease or disorder mentioned herein comprising administering to a subject a pharmaceutically active amount of a compound of Formula (I).
Furthermore, compounds of the Formula (I) are also useful, in combination with one or several immunomodulating agents, for the prevention and/or treatment of the diseases and disorders mentioned herein. According to a preferred embodiment of the invention, said agents are selected from the group consisting of immunosuppressants, corticosteroids, NSAID's, cytotoxic drugs, adhesion molecule inhibitors, cytokines, cytokine inhibitors, cytokine receptor antagonists and recombinant cytokine receptors.
The present invention also relates to the use of a compound of Formula (I) for the preparation of a pharmaceutical composition, optionally for use in combination with

structure 3 with N,0-dimethylhydroxylamine hydrochloride in the presence of a coupling reagent such as EDC, DCC, etc. (M. Mentzel, H. M. R. Hoffmann, N-Methoxy N-methyl amides (Weinreb amides) in modern organic synthesis, Journal fuer Praktische Chemie/Chemiker-Zeitung 339 (1997), 517-524; J. Singh, N. Satyamurthi, I. S. Aidhen, The growing synthetic utility of Weinreb's amide, Journal fuer Praktische Chemie (Weinheim, Germany) 342 (2000) 340-347; V. K. Khiestkin, D. G. Mazhukin, Recent advances in the application of N,0-dialkylhydroxylamines in organic chemistry, Current Organic Chemistry 7 (2003), 967-993).

In case A represents -CO-CH=CH-, the compounds of Formula (I) may be prepared by reacting a compound of Structure 4 with a compound of Structure 5 in the presence of a base or an acid. Compounds of Formula (I) wherein A represents -CO-CH2-CH2- may also be prepared by reacting a compound of Formula (I) wherein A represents -CO-CH=CH- (Structure 6) with hydrogen in the presence of a catalyst such as Pd/C, Pt/C, PtOa, etc. in a solvent such as ethanol, methanol, THF, etc.

Compounds of the Formula (I) wherein A represents -CO-CH2-NH- may be prepared by reacting a compound of Structure 7 with a compound of Structure 8 in the presence or absence of a base such as K2CO3, Na2C03, K-tert.butoxide, NaOH,

NaH, triethylamine, DIPEA, etc. in a solvent such as acetone, DMF, THF, dioxane, etc., or mixtures thereof. The compounds of Structure 7 can be prepared by reacting a compound of Structure 4 with a brominating agent such as phenyltrimethylammoniumbromid dibromlde, benzyltrimethylammonium-tribromid, triphenylphosphine dibromide, etc. in a solvent such as DCM, chloroform, THF, diethyl ether, methanol, ethanol, etc., or mixtures thereof.

A compound of Structure 4 may be prepared by treating a compound of Structure 3 with MeLi in a solvent such as diethyl ether, THF, and dioxane, at temperatures between -20 and 50°C. Alternatively, a compound of Structure 4 may be prepared by reacting a compound of Structure 1 with methylmagnesium bromide.
Compounds of Formula (I) which represent a 5-thiophen-2-yl-[1,2,4]oxadiazole derivative, are prepared by reacting a compound of Structure 9 in a solvent such as xylene, toluene, benzene, pyridine, DMF, dichloromethane, acetic acid, trifluoroacetic acid, etc. at rt or elevated temperatures in the presence or absence of auxiliaries such as acids (e.g. TFA, acetic acid, HCI, etc.), bases (e.g. NaH, NaOAc, NaaCOa, K2CO3, triethylamine, etc.), tetraalkylammonium salts, or water removing agents (e.g. oxalyl chloride, a carboxylic acid anhydride, POCI3, PCI5, P4O10, molecular sieves, etc.) (Lit: e.g. A. R. Gangloff, J. Litvak, E. J. Shelton, D. Sperandio, V. R. Wang, K. D. Rice, Tetrahedron Lett. 42 (2001), 1441-1443; T. Suzuki, K. Iwaoka, N. Imanishi, Y. Nagakura, K. Miyta, H. Nakahara, M. Ohta, T. Mase, Chem. Pharm. Bull. 47 (1999), 120-122; R. F. Poulain, A. L. Tartar, B. P. D6prez, Tetrahedron Lett. 42 (2001), 1495-1498; R. M. Srivastava, F. J. S. Oliveira, D. S. Machado, R. M. Souto-Maior, Synthetic Commun. 29 (1999), 1437-1450; E. 0. John, J. M. Shreeve, Inorganic Chemistry 27 (1988), 3100-3104; B. Kaboudin, K. Navaee, Heterocycles 60 (2003), 2287-2292).

Synthesis, 3"^ Edition, Wiley New York, 1991; P. J. Kocienski, Protecting Groups, Thieme Stuttgart, 1994). Alternatively, the desired residues R^ to R^ may also be introduced in later steps that follow the reaction of a compound of Structure 1, 4, 7, 3, and 11 with a suitable precursor of a compound of Structure 2, 5, 8, 10 and 12, respectively. The compounds of Structure 2, 5, 8, 12, and 13 or their precursors are either commercially available or are prepared according to procedures known to a person skilled in the art.
A compound of Formula (I), wherein R^ represents -(CH2)k-(CHR^^)p-CHR^®-CONR^^R®^ may be prepared by reacting a compound of Formula (III) with the appropriate amine in the presence of a coupling reagent such as TBTU, EDC, etc. The compounds of Formula (III) are prepared in analogy to the procedures outlined above for the preparation of compounds of Formula (I).

A compound of Structure 3 may be prepared by reacting a compound of Structure 15 with an aqueous base such as aq. NaOH, aq. LiOH, aq. KOH, etc. or an acid such as aq. HCI, TFA, etc. in a solvent such as water, ethanol, methanol, THF, etc. or mixtures thereof.

The compounds of Structure 16 may also be obtained by reacting a compound of Structure 18 with e.g. trifluoromethanesulfonic acid anhydride in DCM in the presence of a base to give a compound of Structure 19 (Lit. e.g. G. T. Crisp, A. G. Meyer, J. Org. Chem. 57 (1992) 6972-6975; R. M. Keenan, et al. J. Med. Chem. 35 (1992) 3858-3872) which is then converted to a compound of Structure 16 by treatment with a mercaptoacetic acid ester or mercaptoacetonitrile as described above.
The compounds of Structure 18 may be prepared by acylating a compound of Structure 20 with an appropriate acylating agent such as ethyl or methyl formate, methyl or ethyl acetate, methyl or ethyl propionate, chloroformate, acetyl chloride, etc. in the presence of a base such as K-tert. butylate, NaOMe, NaH, LDA, etc. in a solvent such as THF, toluene, EtOH etc. at temperatures between 0 and 60''C. (Lit. e.g. Ch. Kashima, S. Shibata, H. Yokoyama, T. Nishio, Journal of Heterocyclic Chemistry 40 (2003), 773-782; I. Yavari, Issa, M. Bayat, Tetratiedron 59 (2003), 2001-2005; J. P. Konopelski, J. Lin, P. J. Wenzel, H. Deng, G. I. Elliott, B. S. Gerstenberger, Organic Letters 4 (2002) 4121-4124; C. Wiles, P. Watts, S. J. Haswell, E. Pombo-Villar, Tetrahedron Letters 43 (2002), 2945-2948; R. Faure, A. Frideling, J.-P. Galy, I. Alkorta, J. Elguero, Heterocycles 57 (2002) 307-316; via imine: M. Hammadi, D. Villemin, Synthetic Communications 26 (1996) 2901-2904).


The compounds of Structure 18 may also be prepared by alkylation of the appropriate dicarbonyl compound of Structure 21 under conditions known to a person skilled in the art.
The compounds of Structure 20 and 21 are either commercially available or are prepared according to procedures known to a person skilled In the art. Compounds of Structure 17, wherein R^ represents hydrogen may also be prepared by reacting a compound of Structure 20 containing the desired residue R^ under Vilsmeyer conditions with POCI3/DMF in a solvent such as DCM (e.g. G. Alvernhe, D. Greif, B. Langlois, A. Laurent, I. Le Dr6an, M. Pulst, A. Selmi, M. Weissenfels, Bull. Soc. Chim. Fr. 131 (1994) 167-172).
Examples
The following examples illustrate the invention but do not at all limit the scope thereof.
All temperatures are stated in °C. Compounds are characterized by ^H-NMR (300 MHz) or ^^C-NMR (75 MHz) (Varian Oxford; chemical shifts are given in ppm relative to the solvent used; multiplicities: s = singlet, d = doublet, t = triplet; p = pentuplet, hex = hexet, hept = heptet, m = multiplet, br = broad, coupling constants are given in Hz); by LC-MS (Finnigan Navigator with HP 1100 Binary Pump and DAD, column: 4.6x50 mm, Zorbax SB-AQ, 5 ^m, 120 A, gradient: 5-95% acetonitrile in water, 1 min, with 0.04% trifluoroacetic acid, flow: 4.5 mL/min), XR is given in min; by TLC (TLC-plates from Merck, Silica gel 60 F254); or by melting point. Compounds are purified by preparative HPLC (column: X-terra RP18, 50x19 mm, 5 ^m, gradient: 10-95% acetonitrile in water containing 0.5 % of formic acid) or by MPLC (Labomatic MD-80-100 pump. Linear UVIS-201 detector, column: 350x18 mm, Labogel-RP-18-5s-100, gradient: 10% methanol in water to 100% methanol).

Abbreviations (as used lierein):
aq. aqueous
atm atmosphere
Boc-sarcosine N-tert. butyloxycarbonyl-sarcosine
BSA bovine serum albumin
Bu butyl
CC column chromatography
CD! carbonyl diimidazole
DBU 1,8-diazabicylo[5.4.0]undec-7-en
DCC dicyclohexyl carbodiimide
DCM dichloromethane
DIPEA diisopropyl-ethylamine, HQnig's base, ethyl-diisopropylamine
DMF dimethylformamide
DMSO dimethylsulfoxide
DPPP 1,3-bis-(diphenylphosphino)-propane
EA ethyl acetate
EDC N-(3-dimethylaminopropyl)-N'-ethyl-carbodiimide
Et ethyl
EtOH ethanol
h hour(s)
H BTU 0-(benzotriazol-1 -yl)-N, N, N', N'-tetramethyluronium
hexafluorophosphate
HOBt 1-hydroxybenzotriazole
HPLC high performance liquid chromatography
HV high vacuum conditions
KOtBu potassium tert-butoxide
LC-MS liquid chromatography - mass spectrometry
LDA lithium diisopropyl amide
Me methyl
MeOH methanol
min minute(s)
MPLC medium pressure liquid chromatography
NaOAc sodium acetate

NMO N-methyl-morpholine-N-oxide
OAc acetate
Ph phenyl
prep. preparative
rt room temperature
sat. saturated
S1P sphingosine 1-phosphate
TBTU 2-(1 H-benzotriazole-1-yl)-1,2,3,3-tetramethyluronium
tetrafluoroborate
Tf trifluoromethylsulfonyl
TFA trifluoroacetic acid
THF tetrahydrofuran
TLC thin layer chromatography
tR retention time given in minutes

a) Phosphorusoxychloride (53.7 g, 350 mmol) is slowly added to DMF (60 mL) stirred at 5°C. Upon complete addition, the clear solution is stirred for further 30 min at 5°C before 5-methyl-2-hexanone (20 g, 175 mmol) is added dropwise. The yellow solution is stirred for 30 min at 0°C, then for 90 min at rt. The mixture becomes warm (40°C) and a thick suspension forms. The mixture is cooled to 25°C and stirring is continued for 1 h before it is poured into an aq. solution of NaOAc (80 g)/ice mixture. The mixture is extracted twice with diethyl ether. The organic extracts are washed with water, combined, dried over MgS04, filtered and evaporated to give crude 3-chloro-2-isobutyl-but-2-enal (35.4 g) as a yellow oil, LC-MS: tp = 0.97 min.

b) Sodium (10.7 g, 467 mmol) is dissolved in ethanol (500 mL) and the resulting solution is diluted with THF (100 mL) before mercapto-acetic acid ethyl ester (33.7 g, 280 mmol) dissolved in THF (70 mL) is slowly added at 5°C. The mixture is stirred at rt for 1 h before a solution of 3-chloro-2-isobutyl-but-2-enal (30 g, 187 mmol) in THF (100 mL) is slowly added at 8°C. The resulting yellow suspension is stirred at rt for 16 h. The reaction mixture is diluted with diethyl ether (500 mL) and is washed with dilute aq. NaOCI solution, followed by aq. IN HCI and water. The organic extract is dried over MgS04, filtered and evaporated. The remaining orange oil is dissolved in ethanol (150 mL) and 2N aq. LiOH (50 mL) is added. The mixture is stirred for 16 h at 50°C, acidified with 2 N aq. HCI and extracted with EA. The organic extract is dried over MgS04, filtered and evaporated. The crude product is recrystallized from EA/heptane to give 4-isobutyl-5-methyl-thiophene-2-carboxylic acid (10.5 g) as colourless crystals; LC-MS: tR = 0.92 min, [M+I+CH3CN] = 240.16; ^H NMR (CDCI3): 5 7.59 (s, 1H), 2.40-2.37 (m, 5H), 1.84 (hept, J = 7.0 Hz, 1H), 0.90 (d, J = 7.0 Hz, 6H).

At -78''C, tert.-butyllithium (20 mL, 1.5 M in pentane) is slowly added to a solution of 4-isobutyl-5-methyl-thiophene-2-carboxylic acid (2.0 g, 10.1 mmol) in THF (100 mL). The mixture turns dark. The mixture is stirred at -78°C for 15 min, then a solution of iodoethane (6.18 g, 39.6 mmol) in THF (10 mL) is added. The mixture is stirred for 1 h at -78°C. Further iodoethane (6.18 g, 39.6 mmol) is added and the mixture allowed to warm to rt over a period of 15 h. The reaction is quenched by adding 1 M aq. NaH2P04 (20 mL) and 1 N aq. HCI (50 mL) and the mixture is extracted with chloroform (1x100 mL, 3x30 mL). The combined organic extracts are dried over MgS04, filtered and evaporated. The crude product is purified by MPLC on silica gel eluting with a gradient of EA in hexane to give 3-ethyl-4-isobutyl-5-

methyl-thiophene-2-carboxylic acid (1.29 g) as a yellow solid; LC-MS: tp = 1.00 min, [M+I+CH3CN] = 268.29; 'H NMR (CDCI3): 5 2.92 (q, J = 7.0 Hz, 2H), 2.40 (s, 3H), 2.37 (d, J = 7.6 Hz, 2H), 1.80 (n, J = 7.0 Hz, 1H), 1.15 (t, J = 7.6 Hz, 3H), 0.92 (d, J = 7.0 Hz, 6H).

a) To a solution of KOtBu (50 g, 446 mmol) in THF (400 mL) is added during 30 min
ethylformate (92 g, 1.25 mol). Strong gas evolution occurs. The mixture is cooled
during the addition with a water bath at WC. After complete addition, the mixture is
stirred until the gas evolution ceases (15 min). The mixture is cooled with ice at 0°C
and a mixture of 5-methyl-2-hexanone (34.25 g, 300 mmol) and ethylformate (41 g,
0.55 mol) is added slowly during 30 min. The mixture is stirred for 15 h, diluted with
EA (500 mL) and washed with 1 N aq. HCI (100 mL), 1 M aq. NaH2P04 solution
(100 mL) and brine (100 mL). The organic extract is dried (MgS04), filtered and
evaporated to give crude 4-hydroxy-3-isobutyl-but-3-en-2-one (28 g) which is used
without further purification; LC-MS: tp = 0.80 min, [M+1] = 143.39.
b) To a solution of 4-hydroxy-3-isobutyl-but-3-en-2-one (28 g, 197 mmol) in
chloroform (350 mL) a soution of oxalylchloride (44.3 g, 349 mmol) in chloroform
(50 mL) is added slowly during 5 min. The resulting dark brown mixture is stirred at
rt for 2 h before it is cooled to 0°C and treated with ice (100 g) followed by 1 N aq.
NaOH (100 mL). When the quite violent gas evolution ceases the phases are
separated (the still acidic aq. phase is discarded). The organic phase is washed
with IN aq. NaOH (3 x 75 mL) and 1 N aq. NaH2P04 (75 mL), dried (MgS04),
filtered and evaporated to give crude 4-chloro-3-isobutyl-but-3-en-2-one (31.6 g) as
a dark brown oil; LC-MS: tp = 0.97 min.

c) KOtBu (44.2 g, 394 mmol) is added portionwise to ethanol (200 mL). The mixture Is stirred for 30 min at 20°C to dissolve all KOtBu. Mercapto-acetic acid ethyl ester (47.3 g, 394 mmol) is added and the temperature is maintained at 20''C. This solution is slowly added at 20°C to a solution of the crude 4-chloro-3-isobutyl-but-3-en-2-one (31.6 g, 197 mmol) in THF (350 mL). The mixture is stirred at rt for 15 h before sodium ethylate (13.4 g, 197 mmol) is added and stirring is continued at reflux for 1 h. The mixture is cooled to rt, and the solvents are evaporated on a rotavap. The residue is diluted with diethyl ether (500 mL), washed with 1 M aq. NaH2P04 (200 mL), 1 N aq. NaOH (2x100 mL), sat. aq. NaHCOs (35 mL) containing 10% aq. NaOCI (15 mL) and brine (100 mL), dried over Na2S04, filtered and evaporated. The resulting residue (36.3 g) is dissolved in EtOH (250 mL), 2 N aq. LiOH (100 mL) is added, and the mixture is stirred at rt for 48 h before it is extracted with diethyl ether (1x400 mL, 2x150 mL). The organic extracts are washed with 1 N aq. NaOH (3x100 mL). The aq. extracts are carefully acidified with 25% aq. HCI and then extracted with DCM (3x150 mL). The combined DOM extracts are dried over MgS04, filtered and evaporated. The crude product is purified by crystallisation from acetonitrile (150 mL) at 4''C to give 4-isobutyl-3-methyl-thiophene-2-carboxylic acid (16.0 g) as a beige-brown crystalline powder; LC-MS: tR = 0.95 min; ''H NMR (CD3OD): 5 7.21 (s, 1H), 2.43 (s, 3H), 2.42 (d, J = 7.0 Hz, 2H), 1.83 (n, J = 7.0 Hz, 1H), 0.92 (d, J = 7.0 Hz, 6H).

To a solution of 4-isobutyl-3-methyl-thiophene-2-carboxylic acid (991 mg, 5.0 mmol) in THF (30 mL), tert.-BuLi (7.3 mL, 11 mmol, 1.5 M in pentane) is slowly added at -78°C. The mixture is stirred at -78°C for 1 h, then iodomethane (1.6 mL, 25.7 mmol) is added. The mixture is stirred and is allowed to warm to rt over a period of 15 h before the reaction is quenched with 1 N aq. HCI (50 mL) and extracted with DCM (1x100 mL, 2x50 mL). The combined organic extracts are dried (MgS04),

a) To a slurry of Mg turnings (3.89 g, 160 mmol) in dry diethyl ether (150 mL), a solution of isoamylbromide (24.59 g, 163 mmol) in diethyl ether (20 mL) is slowly added. As soon as the reaction starts, the mixture is cooled if necessary with a water bath. After complete addition, the mixture is stirred for 30 min, and is then added slowly to a cooled (0°C) mixture of isobutyronitrile (34.65 g, 501 mmol) and CuBr (1.15 g, 8.0 mmol) in diethyl ether (50 mL) and THF (50 mL). After complete addition (30 min) the mixture is stirred at rt for 1 h before 1 N aq. NaHaPOA (50 mL) and IN aq. HCI (100 mL) is added. Stirring is continued at rt for 15 min. The phases are separated and the aq. phase is extracted with additional diethyl ether (100 mL). The combined organic extracts are washed with aq. IN aq. HCI (2 x 50 mL), sat. aq. NH4CI/sat. aq. NaHCOa 1:1 (2 x 50 mL) and brine (50 mL), dried over Na2S04, filtered and evaporated to give crude 2,6-dimethyl-heptan-3-one (20.1 g) as an oil.
b) To a solution of KOtBu (15.6 g, 139 mmol) in THF (200 mL) is added ethylformate (27.4 g, 370 mmol). During the addition, the mixture is cooled with a water bath to 10°C. Strong gas evolution occurs. The mixture is stirred until the gas evolution ceases (5 min) and is then cooled to -5°C. To this mixture a solution of 2,6-dimethyl-heptan-3-one (20 g, 141 mmol) and ethylformate (13.7 g, 185 mmol) in THF (20 mL) is added slowly during 30 min. The reaction mixture is stirred for 2 h and allowed to slowly warm to rt. The reaction is diluted with diethyl ether (200 mL) and washed with 1 N aq. HCI (2 x 100 mL). The organic extract is dried (Na2S04), filtered and evaporated to give 1-hydroxy-2-isobutyl-4-methyl-pent-1-en-3-one (7.35 g) as a brown resin; LC-MS: tR = 0.91 min, [M+1] = 171.21.
c) Over a period of 20 min, a solution of oxalylchloride (6.5 mL, 9.6 g, 75.6 mmol) in
chloroform (10 mL) is added slowly to a solution of 1-hydroxy-2-isobutyl-4-methyl-
pent-1-en-3-one (7.35 g, 43.2 mmol) in chloroform (100 mL) cooled to -10°C. The
mixture is stirred for further 30 min, quenched with ice (100 g) and 1 N aq. NaOH

(100 mL). When the quite violent gas evolution ceases the phases are separated. The organic phase is washed with 1 N aq. NaOH (3 x 75 mL) and 1 N aq. NaH2P04 (75 mL), dried (MgS04), filtered and evaporated to give crude 1-chloro-2-isobutyl-4-methyl-pent-1-en-3-one (14.2 g) as a brown oil.
d) Mercapto-acetic acid ethyl ester (9.86 g, 82.1 mmol) is slowly added to a solution of KOtBu (9.50 g, 84.7 mmol) in ethanol (50 mL) cooled to 10-15''C. The mixture is diluted with THF (100 mL) before a solution of the crude 1-chloro-2-isobutyl-4-methyl-pent-1-en-3-one (corresponds to 8.1 g, 43 mmol) in THF (50 mL) is added. The reaction mixture is stirred at 40°C for 15 h, then at reflux for further 20 h, and at 35°C for 48 h. The mixture is diluted with ether (300 mL) and washed with 1N aq. NaOH (3 X 100 mL), 1 M aq. KHSO4 (100 mL) and brine (100 mL). The organic phase is dried (Na2S04), filtered and evaporated. The resulting residue is dissolved in ethanol (30 mL), 2 N aq. LiOH (30 mL) is added, and the mixture is stirred at reflux for 48 h. The mixture is extracted with diethyl ether, then the aq. phase is acidified with 25% aq. HCI and extracted with DCM. The organic extracts are dried (MgS04), filtered and evaporated. The brown oil is diluted in acetonitrile (5 mL) and allowed to crystallize at A'C. The crystalline material is collected, washed with acetonitrile and dried to afford pure 4-isobutyl-3-isopropyl-thiophene-2-carboxylic acid (98 mg) as pale yellow crystals; ^H NMR (CD3OD): 5 7.16 (s, 1H), 3.68 (hept, J = 7.0 Hz, 1H), 2.48 (d, J = 7.6 Hz, 2H), 1.84 (hept, J = 7.0 Hz, 1H), 1.34 (d, J = 7.0 Hz, 6H), 0.92 (d, J = 6.4 Hz, 6H).

Intermediate A1 (10.0 g, 50.4 mmol) is dissolved in CHCI3 (100 mL) and thionylchloride (15 mL) is added at rt. The mixture is stirred at reflux for 2 h. The mixture is evaporated to provide the crude acid chloride (11.2 g). A part of this material (6.5 g, 30 mmol) is dissolved in DCM (200 mL) and then added to a cold (0°C) solution of hydrazine (90 mL, 1 M in THF). The mixture is stirred and warmed to rt over a period of 15 h before it is diluted with diethyl ether (150 mL) and washed with 1M aq. HCI (75 mL, then 5 x 50 mL). The combined aq. extracts are washed with ether (50 mL), basified with 33% aq. KOH and extracted with DCM (5 x 50 mL). The organic DCM extracts are dried over Na2S04, filtered and evaporated to give 4-isobutyl-5-methyl-thiophene-2-carboxylic acid hydrazide (6.24 g) as a white solid; LC-MS: tR = 0.75 min, [M+1] = 213.12; ^H NMR (De-DMSO): 6 0.85 (d, J = 6.7 Hz, 6 H), 1.76 (hept, J = 6.7 Hz, 1H), 2.26-2.34 (m, 5 H), 4.34 (s, 2 H), 7.41 (s, 1 H), 9.52 (s, 1 H).



To a solution of 4-isobutyl-5-methyl-thiophene-2-carboxylic acid (126 mg, 637 |umol) in DCM (5 mL), DIPEA (249 mg, 1.93 mmol) is added followed TBTU (202 mg, 628 ^mol). The mixture is stirred at rt for 30 min before 3-[2-ethyl-4-(N-hydroxycarbamimidoyl)-6-methyl-phenyl]-propionic acid (159 mg, 637 ^mol) is added. The ixture is stirred at rt for 16 h before it is diluted with DCM, washed with 1 N aq. HCI solution, dried over Na2S04, filtered and concentrated. The residue is dissolved in toluene (20 mL) and the reaction mixture is stirred at 110°C for 18 h. The solvent is evaporated and the crude product is purified by CC on silica gel eluting with DCM containing 5% of methanol to give 3-{2-ethyl-4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-6-methyl-phenyl}-propionic acid (41 mg) as a beige solid; LC-MS: tR = 1.17 min, [M+lf = 413.30; ^H NMR (CDCI3): 5 0.94 (d, J = 6.4 Hz, 6 H), 1.30 (t, J = 7.6 Hz, 3 H), 1.84-1.96 (m, 1 H), 2.43 (s, 3 H), 2.45 (s, 3 H), 2.46-2.60 (m, 4 H), 2.75 (q, J = 7.3 Hz, 2 H), 3.02-3.12 (m, 2 H), 7.66 (s, 1 H), 7.79(s, 1H), 7.81 (s, 1 H).

To a solution of 4-(2-tert-butoxycarbonyl-ethyl)-3,5-dimethyl-benzoic acid (763 mg, 3.59 mmol) in DCM (18 mL), DIPEA (542 mg, 4.20 mmol) is added followed by TBTU (1.27 g, 3.95 mmol). The mixture is stirred for 15 min before Intermediate CI (1.0 g, 3.59 mmol) is added. Stirring is continued at rt for 16 h. The mixture Is diluted with diethyl ether, washed with 1 N aq. NaOH solution, then 1 N aq. HCI solution followed by brine, dried over Na2S04, and filtered. The solvent is

evaporated to give 3-{4-[N'-(4-isobutyl-5-methyl-thiophene-2-carbonyl)-hydrazinocarbonyl]-2,6-dimethyl-phenyl}-propionic acid (1.66 g) as a colourless foam; LC-MS: tR = 1.09 min, [M+1] = 473.48. To a solution of this material in THF (15 mL) Burgess reagent (1.26 g, 5.27 mmol) is added. The mixture is heated to 110°C for 3 min in a microwave oven. The mixture is cooled to rt, diluted with diethyl ether and washed with 1 N aq. NaOH. The organic extract is dried over Na2S04, filtered and concentrated to give 3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenyl}-propionic acid methyl ester (1.26 g) as a yellow oil that slowly solidifies; LC-MS: tg = 1.24 min. [M+1] = 455.21. A suspension of this ester in formic acid (20 mL) is stirred at rt for 3 h. The formic acid is evaporated and the residue is purified by CC on silica gel eluting with DCM containing 5% of methanol to give 3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenyl}-propionic acid as a pale yellow solid; LC-MS: tR = 1.07 min, [M+1] = 399.16.



a) 4-lsobutyl-3-methyl-thiophene-2-carbonitrile is obtained by reacting 4-chloro-3-
isobutyi-but-3-en-2-one witli thioacetic acid S-cyanomethyl ester in tlie presence of
NaOEt as described in step c) for Intermediate A3. The crude product is purified by
CC on silica gel eluting with heptane:EA 6:1; LC-MS: tp = 1.03 min; ''H NMR
(CDCI3): 5 0.92 (d, J = 6.4 Hz, 12 H), 1.78-1.90 (m, 1 H), 2.34 (s, 3 H), 2.40 (d, J =
7.3 Hz, 2 H), 7.11 (s, 1 H).
b) To a solution of 4-isobutyl-3-methyl-thiophene-2-carbonitrile (2.10 g, 11.7 mmol)
in methanol (50 mL), NaHCOa (1.38 g, 16.4 mmol) followed by hydroxylamine
hydrochloride (977 mg, 14.1 mmol) is added. The mixture is stirred at 60°C for 20 h
before it is filtered. The solvent of the filtrate is evaporated and the remaining

times with diethyl ether. The combined organic extracts are dried over Na2S04, filtered and concentrated. The crude product is purified by CC on silica gel eluting with heptane:EA 4:1 to give 3-chloro-4-hydroxy-5-methyl-benzaldehyde (2.79 g) as a beige solid, LC-MS; tp = 0.82 min.
b) 3-Chloro-4,N-dihydroxy-5-methyl-benzamidine is prepared from the above aldehyde following literature procedures (A. K. Chakraborti, G. Kaur, Tetrahedron 55 (1999) 13265-13268; E. Meyer. A. C. Joussef, H. Gallardo, Synthesis 2003, 899-905), LC-MS: IR = 0.48 min, [M+1] = 201.17; ^H NMR (CD3OD): 5 2.21 (s, 3 H), 7.22 (d, J = 2.1 Hz, 1 H), 7.37 (d, J = 2.1 Hz, 1 H).

3-Chloro-4,N-dihydroxy-5-methoxy-benzamidine is prepared from 3-chloro-4-hydroxy-5-methoxy benzaldehyde following literature procedures (A. K. Chakraborti, G. Kaur, Tetrahedron 55 (1999) 13265-13268; E. Meyer, A. C. Joussef, H. Gallardo, Synthesis 2003, 899-905), LC-MS: tp = 0.48 min, [M+l] = 217.21; ^H NMR (CD3OD): 5 3.90 (s, 3 H), 7.16 (d, J = 1.8 Hz, 1H), 7.23 (d, J = 1.8 Hz, 1 H).

a) To an ice-cooled solution of 4-hydroxy-3,5-dimethyl-benzoic acid methyl ester (7,52 g, 41.7 mmol) in DCM (250 mL) and pyridine (10 mL), trifluoromethanesulfonic acid anhydride (13.0 g, 45.9 mmol) is added over a period of 20 min. Upon complete addition, the ice bath is removed and the reaction is stirred for further 1 h at rt. The mixture is diluted with DCM (150 mL), washed with

10% aq. citric acid solution followed by brine, dried over MgS04, filtefed and evaporated. The residue is purified by flash chromatography on silica gel eluting with heptane; EA 9:1 to give 3,5-dimethyl-4-trifluoromethanesulfonyloxy-ben2oic acid methyl ester (11.8 g) as colourless fine needles; LC-MS: tp = 1.08 min.
b) To a stirred solution of the above triflate (11.8 g, 37.8 mmol) in dry DMF (155 mL) is sequentially added triethylamine (7.6 g, 75.6 mmol), tert.-butyl acrylate (48.4 g, 378 mmol), DPPP (779 mg, 1.89 mmol) and Pd(0Ac)2 (424 mg, 1.89 mmol) under nitrogen. The mixture is stirred at 115°C for 18 h before another portion of DPPP (160 mg, 0.39 mmol) and Pd(0Ac)2 (80 mg, 0.36 mmol) is added. Stirring is continued for 4 h at IIS'C before the mixture is cooled to rt, diluted with diethyl ether (350 mL) and washed with 1 N aq. HCI, followed by sat. aq. NaHCOs solution. The organic extract is dried over MgS04, filtered and evaporated. The residue is purified by flash chromatography on silica gel eluting with heptane: EA 4:1 to give 4-(2-tert-butoxycarbonyl-vinyl)-3,5-dimethyl-benzoic acid methyl ester (11.21 g) as a colourless solid; LC-MS: tp = 1.09 min.
c) To a solution of 4-(2-tert-butoxycarbonyl-vinyl)-3,5-dimethyl-benzoic acid methyl ester (11.2 g, 38.6 mmol) in ethanol (50 mL) and THF (50 mL), Pd/C (1.0 g, 10% Pd) is added. The mixture is stirred for 16 h at rt under 2.5 bar of Ha. The catalyst is filtered off and the filtrate is concentrated and dried under HV to give 4-(2-tert-butoxycarbonyl-ethyl)-3,5-dimethyl-benzoic acid methyl ester (10.8 g) as a colourless oil; LC-MS: tp = 1.08 min.
d) To a solution of 4-(2-tert-butoxycarbonyl-ethyl)-3,5-dimethyl-benzoic acid methyl ester (10.8 g, 37.0 mmol) in ethanol (100 mL) a 2 M aq. solution of LiOH (50 mL) is added at 0°C. The turbid mixture is stirred at 0°C for 30 min, then at rt for 4 h. The mixture is diluted with 10% aq. citric acid solution and extracted three times with diethyl ether. The combined organic extracts are dried over MgS04, filtered and concentrated. The solid residue is suspended in diethyl ether /heptane, stirred at rt, and filtered. The slurry procedure in diethyl ether/heptane is repeated. The solid material is collected and dried under HV to give 4-(2-tert-butoxycarbonyl-ethyl)-3,5-dimethyl-benzoic acid (5.09 g) as a white crystalline powder; LC-MS: tp = 0.95 min.


a) To an ice-cooled solution of 5-ethyl-4-hydroxy-3-methylbenzaldehyde (10.0 g,
60.9 mmol) in DCM (50 mL) and pyridine (15 mL), trifluorometlianesulfonic acid
anhydride (18.9 g, 67 mmol) is added over a period of 20 min. Upon complete
addition, the ice bath is removed and the reaction is stirred for further 2 h at rt. The
mixture is diluted with DCM (150 mL), washed three times with water, dried over
MgS04, filtered and evaporated. The residue is purified by flash chromatography on
silica gel eluting with heptane: EA 9:1 to give trifluoro-methanesulfonic acid 2-ethyl-
4-formyl-6-methyl-phenyl ester (10.75 g) as a pale yellow oil; LC-MS: tp = 1.07 min;
^H NMR (CDCI3): 5 9.98 (s, 1H), 7.70 (s, 1H), 7.66 (s, 1H), 2.85 (q, J = 10.1 Hz,
2H), 2.48 (s, 3H), 1.30 (t, J = 10.2 Hz, 3H).
b) To a stirred solution of the above triflate (10.7 g, 36.1 mmol) in dry DMF (75 mL)
Is sequentially added triethylamine (7.3 g, 72.2 mmol), methyl acrylate (31.1 g, 361
mmol), DPPP (819 mg, 1.99 mmol) and Pd(0Ac)2 (405 mg, 1.81 mmol) under

nitrogen. The mixture is stirred at 115°C for 5 h, cooled to rt, diluted with diethyl ether (350 mL) and washed twice with 1 N aq. HCI and once with sat. aq. NaHCOs solution. The organic extract is dried over MgS04, filtered and evaporated. The residue is purified by flash chromatography on silica gel eluting with heptane: EA 19:1 to give 3-(2-ethyl-4-formyl-6-methyl-phenyl)-acrylic acid methyl ester (5.93 g) as a colourless liquid; LC-MS: tp = 0.99 min.
c) A suspension of 3-(2-ethyl-4-formyl-6-methyl-phenyl)-acrylic acid methyl ester (5.93 g, 25.53 mmol) in methanol (140 mL) and 2 N aq. NaOH (45 mL) is stirred at rt for 1 h. The methanol is evaporated and the aq. solution is extracted twice with DCM. The aq. layer is acidified with 37% aq. HCI. The precipitate that forms is collected, washed with water and dried. The product is further purified by recrystallisation from EA (100 mL) to give 3-(2-ethyl-4-formyl-6-methyl-phenyl)-acrylic acid (4.2 g) as yellow crystals; LC-MS: tp = 0.87 min.
d) To a solution of 3-(2-ethyl-4-formyl-6-methyl-phenyl)-acrylic acid (2.75 g, 12.6 mmol) and DIPEA (1.8 g, 13.8 mmol) in ethanol (80 mL), Pd/C (275 mg, 10% Pd, moistened with 50% water) is added. The mixture is stirred for 16 h at rt under 1 atm of H2. The catalyst is filtered off and the filtrate is concentrated. The residue is dissolved in EA, washed with 2 N aq. HCI, followed by 1 N aq. HCI and brine. The organic extract is dried over Na2S04, filtered and evaporated to give 3-(2-ethyl-4-hydroxymethyl-6-methyl-phenyl)-propionic acid (2.8 g) as a white solid; LC-MS: tp = 0.76 min.
e) A solution of 3-(2-ethyl-4-hydroxymethyl-6-methyl-phenyl)-propionic acid (2.8 g, 12.6 mmol) in acetic acid (50 mL) is treated with Mn02 (3.9 g, 45.4 mmol) and the resulting mixture is stirred at 80°C for 4 h. The mixture is filtered and the filtrate is concentrated. The crude product is purified by CC on silica gel eluting with DCM to give 3-(2-ethyl-4-formyl-6-methyl-phenyl)-propionic acid (1.76 g) as a beige solid; LC-MS: tR = 0.86 min.
f) A solution of 3-(2-ethyl-4-formyl-6-methyl-phenyl)-propionic acid (1.67 g, 7.58 mmol) and hydroxylamine hydrochloride (780 mg, 11.36 mmol) in 1-methyl-2-

pyrrolidone is heated to 80°C for 30 min in the microwave (300 W, active cooling during irradiation). The reaction mixture is diluted with diethyl ether and washed with water and brine. The organic extract is dried over Na2S04, filtered and evaporated to give 3-(4-cyano-2-ethyl-6-methyl-phenyl)-propionic acid (1.55 g) as a beige solid; LC-MS: tp = 0.89 min, ''H NMR (De-DMSO): 5 12.25 (s, 1H). 7.45 (s, 2H), 2.91-2.84 (m, 2H), 2.67-2.59 (m, 2H), 2.35-2.30 (m, 5H), 1.14 (t, J = 7.6 Hz, 3H).
g) Potassium tert. butoxide (2.71 g, 24.1 mmol) is carefully dissolved in methanol (25 mL). To this solution hydroxylamine hydrochloride (1.44 g, 20.7 mmol) followed by 3-(4-cyano-2-ethyl-6-methyl-phenyl)-propionic acid (1.50 g, 6.90 mmol) dissolved in methanol (7.5 mL) is added. The mixture is refluxed for 8 h and the solvent is evaporated. The residue is dissolved in 2 N aq. HCI and extracted with EA. The pH of the aq. phase is adjusted to pH 5 by adding sat. aq. NaHCOa and the mixture is extracted three times with EA. The combined organic extracts are dried over Na2S04, filtered, evaporated and dried to give 3-[2-ethyl-4-(N-hydroxycarbamimidoyl)-6-methyl-phenyl]-propionic acid (1.4 g) as a white solid; LC-MS: tR = 0.60 min, [M+lf = 251.17.

4-Benzyioxy-3,5-dimethyl-benzoic acid (5.37 g, 20.9 mmol) is dissolved in CHCI3 (75 mL) and thionylchloride (10 mL) is added at rt. The mixture is stirred at reflux for 2 h. The mixture is evaporated to provide the crude acid chloride (5.62 g). A part of this material (2.75 g, 10 mmol) is dissolved in THF (10 mL) and cooled to -78°C before it is treated with hydrazine (25 mL, 1 M solution in THF). The mixture is warmed to rt over a period of 15 h. The mixture is diluted with ether (150 mL) and washed with 1M aq. HCI (75 mL, then 5 x 50 mL). The combined aq. extracts are washed with ether (50 mL), basified with 33% aq. KOH and extracted with DCM (5

X 50 mL). The DCM extracts are dried over Na2S04, filtered and evaporated to give 4-benzyioxy-3,5-dimetliyl-benzoic acid hydrazide (1.29 g) as a white solid, LC-MS: tR = 0.78 min, [M+lf = 271.19; ^H NMR (CDCI3): 5 2.30 (s, 6 H), 3.86 (s br, 2H), 4.82 (s, 2 H). 7.33-7.49 (m, 7 H), 7.56 (s br, 1 H).

a) 3-Ethyl-4-hydroxy-5-methyl-benzaldehyde is prepared from commercially
available 2-ethyl-6-methyl-phenol following literature procedures (G. Trapani, A.
Latrofa, M. Franco, C. Altomare, E. Sanna, M. Usala, G. Biggio, G. Liso, J. Med.
Chem. 41 (1998) 1846-1854; A. K. Chakraborti, G. Kaur, Tetrahedron 55 (1999)
13265-13268; E. Meyer, A. C. Joussef, H. Gallardo, Synthesis 2003, 899-905); ^H
NMR (CDCI3): 5 9.83 (s, 1H), 7.58-7.53 (m, 2H), 5.30 (s br, 1H), 2.69 (q, J = 7.6 Hz,
2H), 2.32 (s, 3H), 1.28 (t, J = 7.6 Hz, 3H).
b) To a suspension of K2CO3 (21 g, 152 mmol) in acetone (200 mL), 3-ethyl-4-
hydroxy-5-methyl-benzaldehyde (5.0 g, 30.5 mmol) followed by benzyl bromide
(7.87 g, 45.7 mmol) is added. The suspension is refluxed for 16 h before it is
filtered. The filtrate is concetrated and the crude product is purified by CC on silica
gel eluting with heptane:EA 4:1 to give 4-benzyloxy-3-ethyl-5-methyl-benzaldehyde
(5.04 g) as colourless oil; LC-MS: tp = 1.09 min, [M+lf = 255.25.
c) To a solution of 4-benzyloxy-3-ethyl-5-methyl-benzaldehyde (5.0 g, 19.7 mmol)
in acetone (200 mL), KMn04 (4.66 g, 29.5 mmol) is added. The dark violet solution
becomes slightly warm and turns dark brown. The mixture is stirred at rt for 90 min
before the solvent is evaporated. The residue is treated with 10% aq. citric acid
soluiton (200 mL) and brine (200 mL) and extracted four times with DCM (4x200
mL). The combined organic extracts are dried over MgS04, filtered and
concentrated. The crude product is purified by CC on silica gel eluting with

To a solution of 3-(4-hydroxy-3,5-dimethyl-phenyl)-1-(4-isobutyl-3-propyl-thiophen-2-yl)-propan-1-one (108 mg, 0.301 mmol) in isopropanol (5 mL), 3 N aq. NaOH (3 mL) followed by (S)-3-chloro-1,2-propanediol (167 mg, 1.51 mmol) is added. The mixture is stirred at 70°C for 4 h. After 4, 5, 6, 7 and 8 h an additional portion of (S)-3-chloro-1,2-propanediol (5x97 mg, 0.878 mmol) is added. After the last addition, stirring is continued at SO'C for 16 h. The mixture is diluted with water and extracted with diethyl ether. The organic extract is dried over MgS04, filtered and evaporated. The crude product is purified by prep. HPLC to give 3-[4-((S)-2,3-dihydroxy-propoxy)-3,5-dimethyl-phenyl]-1-(4-isobutyl-3-propyl-thiophen-2-yl)-propan-1-one (18 mg) as an almost colourless resin; LC-MS: tp = 1.09 min, [M+1] = 433.40; ^H NMR (CDCI3): 5 7.08 (s, 1H), 6.87 (s, 2H), 4.11-4.03 (m, 1H), 3.90-3.70 (m, 4H), 3.14-3.06 (m, 2H), 2.94-2.82 (m, 4H), 2.60 (s br, 2H), 2.40 (d, J = 7.0 Hz, 2H), 2.24 8s, 6H), 1.90-1.78 (m, 1H), 1.56-1.46 (m, 2H), 1.00 8t, J = 7.0 Hz, 3H), 0.93 (d, J = 6.4 Hz, 6H).

a) A mixture of acetylacetone (2.5 g, 25 mmol), K2CO3 (3.45 g, 25 mmol) and 3-
bromo-2-methylpropene (2.5 mL, 25 mmol) in acetone (80 mL) is stirred at 50°C for
3 days. The mixture is filtered, the filtercake washed with few acetone and the
filtrate evaporated to give 2.48 g of 3-(2-methyl-allyl)-pentane-2,4-dione as a dark
oil that is used without further purification in subsequent steps; LC-MS: tR = 0.87
min, [M+ir= 155.26.
b) A solution of 3-(2-methyl-allyl)-pentane-2,4-dione (2.48 g, 16 mmol) in DCM (10
mL) is cooled at -78°C and DIPEA (17 mL, 98 mmol) is slowly added. The mixture
is stirred for 0.5 h at -78''C, then a solution of trifluoromethanesulfonic anhydride
(3.18 mL, 19.3 mmol) in DCM (15 mL) is slowly added keeping the temperature at

below -65°C. The red-brown solution is stirred for 0.5 h at -78°C and then quenched with ice (50 g) and diethyl ether (200 mL). The phases are separated, the organic phase washed with 1M aq. KHSO4 solution and brine, dried with Na2S04, filtered and evaporated. The residue is purified by reversed phase MPLC (H20-MeOH gradient) to give 1.16 g of trifluoromethanesulfonic acid 2-acetyl-1,4-dimethyl-penta-1,4-dienyl ester; LC-MS: tp = 1.03 min, [M+1]* = 287.01.
c) Sodium hydride (60% in petrolether, 404 mg, 10 mmol) is washed with dry pentane (3x10 mL), then dry THF (30 mL) is added and the suspension is cooled at 0°C. A solution of mercaptoacetic acid ethyl ester (0.445 mL, 4 mmol) in THF (4 mL) is slowly added. After stirring for 0.5 h at 0°C a solution of trifluoromethanesulfonic acid 2-acetyl-1,4-dimethyl-penta-1,4-dienyl ester (1.16 g, 4 mmol) in THF (4 mL) is slowly added. The mixture is stirred at 0°C for 0.5 h, then slowly warmed to rt during 15 h. The reaction mixture is partioned between H2O (75 mL) and DCM (100 mL), the aq. phase is basified with 1M aq. NaOH (50 mL) and extracted with DCM (50 mL). The combined organic extracts are dried (Na2S04), filtered and evaporated. The crude product is purified by reversed phase MPLC (HaO-MeOH gradient) to give 450 mg of 3,5-dimethyl-4-(2-methyl-allyl)-thiophene-2-carboxylic acid ethyl ester; LC-MS: tp = 1.10 min, [M+1]* = 239.05.
d) To a solution of 3,5-dimethyl-4-(2-methyl-allyl)-thiophene-2-carboxylic acid ethyl ester (978 mg, 4.1 mmol) in ethanol (20 mL) is added 3M aq. LiOH (10 mL) and the mixture is stirred at 60°C for 1 h. The mixture is partitioned between DCM (100 mL) and 1M aq. HCI (75 mL). The aq. phase is re-extracted with DCM (2 x 50 mL). The combined organic extracts are dried (MgS04), filtered and evaporated to give 653 mg of 3,5-dimethyl-4-(2-methyl-allyl)-thiophene-2-carboxylic acid as a light yellow powder; LC-MS: \R = 0.94 min, [M+l+CHsCNr = 252.04; ^H NMR (CDCI3): 5 11.50 (br. s, 1H), 4.75 (s, 1H). 4.39 (s, 1H), 3.17 (s, 2H), 2.42 (s, 3H), 2.37 (s, 3H), 1.75 (s, 3H).
e) To a solution of 3,5-dimethyl-4-(2-methyl-allyl)-thiophene-2-carboxylic acid (475 mg, 2.26 mmol) in diethyl ether (15 mL), a solution of methyllithium (3.0 mL, 1.6 M in diethyl ether) is added at rt. The mixture is stirred at rt for 1 h before another

portion of methyllithium (0.45 mL) is added. Stirring is continued for 1 h. The reaction is quenched by the addition of 1 N aq. NaH2P04 solution. The mixture is diluted with diethyl ether, washed with 1 N aq. NaOH, dried over Na2S04 and evaporated to give 1-[3,5-dimethyl-4-(2-methyl-a!lyl)-thiophen-2-yl]-ethanone (395 mg) as a yellow oil; LC-MS: tR = 1.03 min, [M+1] = 209.14.
f) A solution of 1-[3,5-dimethyl-4-(2-methyl-allyl)-thiophen-2-yl]-ethanone (395 mg,
1.9 mmol) and 4-hydroxy-3,5-dimethyl-benzaldehyde (427 mg, 2.84 mmol) in
ethanol (10 mL) and 5 N HCI in isopropanol (5 mL) is stirred at rt for 2 h. The dark
solution is diluted with diethyl ether (100 mL) and washed with a mixture of sat. aq.
NaHCOa and 1M aq. NaOH (1:1, 3 x 35 mL). The organic phase is dried (Na2S04),
filtered and evaporated. The residue is purified by reversed phase MPLC (H2O-
MeOH) to give 1-[3,5-dimethyl-4-(2-methyl-allyl)-thiophen-2-yl]-3-(4-hydroxy-3,5-
dimethyl-phenyl)-propenone (661 mg) as a yellow solid; LC-MS: tn = 1.13 min,
[M+1] = 341.15.
g) A solution of 1-[3,5-dimethyl-4-(2-methyl-allyl)-thiophen-2-yl]-3-(4-hydroxy-3,5-
dimethyl-phenyl)-propenone (38.5 mg, 0.11 mmol) in methanol (5 mL) is treated
with Pd/C (10 mg, 10% Pd) and the resulting slurry is stirred at rt for 2 h under 1 bar
of H2. The catalyst is filtered off and the filtrate purified by TLC (Si02, EA-heptane)
to give 3-(4-hydroxy-3,5-dimethyl-phenyl)-1-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-
propan-1-one (18 mg) as a yellow oil; LC-MS: tR = 1.14 min, [M+lf = 345.12; ""H
NMR (CDCI3): 6 6.78 (s, 2H), 2.97 (t, J = 7.4 Hz, 2H), 2.82 (t, J = 7.4 Hz, 2H), 2.39
(s, 3H), 2.32-2.28 (m, 5H), 2.15 (s, 6H), 0.84 (d, J = 6.4 Hz, 6H).



To a solution of 3-(4-hyclroxy-3,5-dimethyl-phenyl)-1-(4-isobutyl-3-methyl-thiophen-2-yl)-propan-1-one (317 mg, 0.96 mmol) in isopropanol (10 mL), 3 N aq. NaOH (2.5 mL) followed by epichlorohydrine (284 mg, 3.07 mmol) is added. The dark red mixture is stirred at rt for 16 h before it is diluted with sat. aq. NaHCOs and extracted twice with diethyl ether. The organic extracts are washed with water, dried over NaaSOA, filtered and evaporated to give crude 3-(3,5-dimethyl-4-oxiranylmethoxy-phenyl)-1-(4-isobutyl-3-methyl-thiophen-2-yl)-propan-1-one (339 mg) as a yellow oil. The material is dissolved in 7 N NH3 in methanol (7.5 mL) and the resulting solution is stirred in an autoklave at 65°C for 2.5 h. The solvent is removed in vacuo and the residue is dried under HV to leave rac-3-[4-(3-amino-2-hydroxy-propoxy)-3,5-dimethyl-phenyl]-1-(4-isobutyl-3-methyl-thiophen-2-yl)-propan-1-one (353 mg) as an orange oil; LC-MS: tp = 0.88 min, [M+1] = 404.20; ^H NMR (CDCI3): 5 7.06 (s, 1H), 6.88 (s, 2H), 3.98-3.90 (m 1H), 3.85-3.73 (m, 2H),

rac-N-(3-{4-[3-(3,4-Diisobutyl-thiophen-2-yl)-3-oxo-propyl]-2,6-dimethyl-phenoxy}-2-hydroxy-propyl)-2-hyclroxy-acetamide is prepared from rac-3-[4-(3-amino-2-
hydroxy-propoxy)-3,5-dimethyl-phenyl]-1-(3,4-diisobutyl-thiophen-2-yl)-propan-1-one in analogy to Example 17; LC-MS: tn = 1.07 min, [M+1] = 504.35.

a) A solution of 4-isobutyl-3-methyl-thiophene-2-carboxylic acid (1.78 g, 9.0 mmol),
DIPEA (3.78 g, 29.2 mmol) and TBTU (3.21 g, 10.0 mmol) in DMF (20 mL) is stirred
at rt for 15 min before 4,N-dihydroxy-3,5-dimethyl-benzamidine (1.80 g, 10.0 mmol)
is added. The mixture is stirred for 15 h at rt. The solvent is evaporated and the
residue taken up in EA (100 mL) and sat. aq. Na2C03 (100 mL). The phases are
separated and the aq. phase is extracted with additional EA (2 x 75 mL). The
combined organic extracts are washed with sat. aq. Na2C03 (50 mL) and brine (50
mL), dried (Na2S04), filtered and evaporated. The brown residue is suspended in
CHCI3 (50 mL) and filtered. The solid is washed with CHCI3 (25 mL) and dried to
give 4-isobutyl-3-methyl-thiophene-2-carboxylic acid (4,N-dihydroxy-3,5-dimethyl-
benzamidine) ester (2.34 g) as a beige powder; LC-MS: tp = 1.02 min, [M+1] =
361.49; ^H NMR (De-DMSO): 5 8.66 (s br, 1H), 7.45 (s, 1H), 7.33 (s, 2H), 6.43 (s br,
2H), 2.42-2.39 (m, 5H), 2.18 (s, 6H), 1.80 (hept, J = 7.0 Hz, 1H), 0.88 (d, J = 7.0
Hz, 6H).
b) 4-lsobutyl-3-methyl-thiophene-2-carboxylic acid (4,N-dihydroxy-3,5-dimethyl-
benzamidine) ester (1.85 g, 5.13 mmol) is suspended in dry toluene and the
mixture is heated at reflux in a Dean-Stark apparatus for 15 h. The mixture is
filtered and the filtrate is evaporated. The residue is purified by CC on silica gel
eluting with a gradient of EA in heptane to give 4-[5-(4-isobutyl-3-methyl-thiophen-
2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenol (668 mg) as a light yellow powder;
LC-MS: tR = 1.16 min, [M+1] = 343.19; ^H NMR (CDCI3): 6 7.79 (s, 2H), 7.17 8s,


a) To a solution of 4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadia20l-3-yl]-
2,6-dimethyl-phenol (515 mg, 1.50 mmol) in isopropanol (20 mL), 3 N aq. NaOH (5
mL) followed by epichlorohydrine (473 mg, 5.12 mmol) is added and the mixture is
stirred at rt for 15 h. Another portion of epichlorohydrine (473 mg, 5.12 mmol) is
added and stirring is continued for 24 h. The mixture is diluted with sat. aq. Na2C03
and is then extracted with DCM (4x75 mL). The combined organic extracts are
dried (Na2S04), filtered and evaporated. The residue is purified by CC on silica gel
eluting with heptane:EA 4:1 to give rac-3-(3,5-dimethyl-4-oxiranylmethoxy-phenyl)-
5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazole (382 mg) as a white powder;
LC-MS: tR = 1.22 min, [M+1] = 399.28.
b) A suspension of rac-3-(3,5-dimethyl-4-oxlranylmethoxy-phenyl)-5-(4-isobutyl-3-
methyl-thiophen-2-yi)-[1,2,4]oxadJazoie (382 mg, 0.959 mmol) in 7 N NH3 in MeOH
(10 mL) and THF (5 mL) is stirred at 60°C for 15 h. The mixture is diluted with 1M
aq. NaOH (30 mL) and extracted with DCM (4 x 75 mL). The combined organic
extracts are dried (Na2S04), filtered and evaporated to give rac-1-amino-3-{4-[5-(4-
isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-
propan-2-ol (419 mg) as a pale yellow solid; LC-MS: tp = 0.90 min, [M+1] = 416.42.


rac-2-Hyclroxy-N-(2-hydroxy-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-climethyl-phenoxy}-propyl)-acetannicle is prepared from rac-1-annino-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-propan-2-ol in analogy to Example 17; LC-MS: IR = 1.04 min, [M+1] = 474.19; ^H NMR (De-DMSO): 6 7.71 (s, 2H), 7.68 (t br, J = 6 Hz, 1H), 7.62 (s, 2H), 5.54 (t, J = 5 Hz, 1H), 5.28 (d, J = 5.3 Hz, 1H), 3.96-3.88 (m, 1H), 3.81 (d, J = 5 Hz, 2H), 3.79-3.66 (m, 2H), 3.46-3.36 (m, 1H), 3.28-3.17 (m, 1H), 2.58 (s, 3H), 2.31 (s, 6H), 1.85 (hept, J = 7.0 Hz, 1H), 0.90 (d, J = 6.4 Hz, 6H).

A solution of rac-1-amino-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-
[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-propan-2-ol (100 mg, 0.241 mmol) in DCM (1 mL) is added to a stirred solution of Boc-sarcosine (60 mg, 0.317 mmol), DIPEA (38 mg, 0.292 mmol) and TBTU (85 mg, 0.265 mmol) in DCM (5 mL). The mixture is stirred at rt for 1 h, diluted with DCM (50 mL) and washed with sat. aq. NaaCOa (2 x 20 mL). The organic extract is dried (Na2S04), filtered and evaporated. The residue dissolved in 4M HCI in dioxane (10 mL) and stirred for 1 h. The mixture is diluted with DCM (75 mL) and washed with 1 N aq. NaOH (50 mL). The aq. phase is extracted with DCM (3 x 30 mL). The combined organic extracts are dried (Na2S04), filtered and evaporated. The residue is purified on prep. TLC plates with DCM containig 10% of 7 N NH3 in MeOH to give rac-N-(2-hydroxy-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-2,6-dimethyl-phenoxy}-propyl)-2-methylamino-acetamide (89 mg) as a white powder; LC-MS: IR = 0.89 min, [M+1] = 487.26; ^H NMR (De-DMSO): 5 7.85 (t, J = 6 Hz, 1H), 7.70 (s, 2H), 7.65 (s, 1H), 5.27 (d br, J = 5 Hz, 1H), 3.93-3.85 (m, 1H), 3.76-3.65 (m, 2H), 3.43-3.35 (m, 1H), 3.24-3.14 (m, 1H), 3.02 (s, 2H), 2.54 (s, 3H), 2.45 (d, J = 6.4 Hz, 2H), 2.30 (s, 6H), 2.20 (s, 3H), 1.83 (hept, J = 7.0 Hz, 1H), 0.88 (d, J = 7.0 Hz, 6H).

a) 4-Allyloxy-3,5-dimethyl-benzoic acid (Moffett, R.B.; Seay, P.H.; J. Med. Pharm.
Chem. (1960) 2201-2212) (620 mg, 3 mmol), tert.-butylcarbazate (475 mg, 3.6
mmol) and NEta 0.5 mL) are dissolved in DMF (10 mL) and TBTU (1.15 g, 3.6
mmol) is added at 0°C. The mixture is stirred for 2 h at 0°C to rt, quenched into
ether (200 mL) and washed with IIVI aq. HCI (2 x 50 mL), 1M aq. NaOH (2 x 50 mL)
and brine (50 mL). The organic phase is dried (l\/lgS04), filtered and evaporated.
The residue is dissolved in dioxane (10 mL) and 4M HCI in dioxane (4 mL) is
added. The mixture is stirred at rt for 15 h; the mixture is diluted with dry diethyl
ether (10 mL), the precipitate filtered, washed with dry diethyl ether (10 mL) and
dried in vacuo to give 487 mg of 4-allyloxy-3,5-dimethyl-benzoic acid hydrazide
hydrochloride as a white powder; LC-MS: tR = 0.69 min, [M+1]* = 221.21.
b) A solution of 4-isobutyl-3-methyl-thiophene-2-carboxylic acid (100 mg, 0.5
mmol), 4-allyloxy-3,5-dimethyl-benzoic acid hydrazide hydrochloride (192 mg, 0.75
mmol) and NEta (0.28 mL) in DMF (10 mL) is cooled at 0°C and TBTU (200 mg, 0.6
mmol) is added. The mixture is stirred for 15 h while warming from O^C to rt. The
mixture is diluted with diethyl ether (50 mL) and washed with 1M aq. NaOH (2 x 20
mL), 1M aq, HCI (2 x 20 mL) and brine (10 mL). The organic phase is dried
(Na2S04), filtered and evaporated to give 160 mg of 4-allyloxy-3,5-dimethyl-benzoic
acid N'-(4-isobutyl-3-methyl-thiophene-2-carbonyl)-hydrazide; LC-MS: tR = 1.05
min, [M+ir = 400.83.
c) A solution of 4-allyloxy-3,5-dimethyl-benzoic acid N'-(4-isobutyl-3-methyl-
thiophene-2-carbonyl)-hydrazide (160 mg, 0.4 mmol) and (methoxycarbonyl-
sulfamoyl)triethylammonium hydroxide (350 mg, 1.27 mmol) in dry THF (5 mL) is
heated at 110°C in a microwave oven for 6 min. The mixture is poured into diethyl
ether (10 mL) and washed with 1M aq. HCI (10 mL). The aq. phase is re-extracted
with ether (10 mL). The combined organic extracts are dried (Na2S04), filtered and

evaporated to give 181 mg of crude 2-(4-allyloxy-3,5-dimethyl-phenyl)-5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,3,4]oxadiazole; LC-MS: tp = 1.23 min, [M+lf = 383.35.
d) To a solution of 2-(4-allyloxy-3,5-dimethyi-phenyl)-5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,3,4]oxadiazoie (181 mg crude, 0.4 mmol) in a mixture of acetone and water (15:1.5 mL) are added N-methylmorplioline-N-oxide (250 mg, 1.85 mmol) and, subsequently, 2.5 % OSO4 in tert.-butanol (0.1 mL). The mixture is stirred at rt for 36 h. The mixture is quenched into diethyl ether (75 mL) and washed with 1M aq. NaOH (50 mL), 1M aq. HCI (30 mL) and brine (20 mL). The organic phase is dried (Na2S04), filtered and evaporated. The residue is purified by TLC (Si02, EA-heptane) to give 118 mg of 3-{4-[6-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenoxy}-propane-1,2-diol as a white powder; LC-MS: tR = 1.00 min, [M+lf = 417.28; ^H NMR (De-DMSO): S, 7.70 (s, 2 H) 7.51 (s, 1 H), 4.96 (d, J = 5.0 Hz, 1 H), 4.63 (t, J = 5.6 Hz, 1 H), 3.68-3.89 (m, 3 H), 3.47 (t, J = 5.6 Hz, 2 H), 2.51 (s, 3 H), 2.45 (m, 2 H), 2.33 (s, 6 H), 1.85 (m, 1 H), 0.90 (d, J = 6.7 Hz, 6 H).

The mixture is stirred at rt for 15 h before it is diluted with diethyl ether (100 mL)
and washed with 1 N aq. NaOH (2x30 mL), 1 M aq. KHSO4 (30 mL) and brine (30
mL). The organic extract is dried (Na2S04), filtered and evaporated. The yellow oily
residue is dissolved in dry toluene (10 mL) and heated at 150°C for 20 min in a
microwave oven. The mixture is poured into diethyl ether (100 mL) and washed
with brine (75 mL). The organic extract is dried (Na2S04), filtered and evaporated to
give 3-(4-allyloxy-2-methoxy-phenyl)-5-(4-isobutyl-5-methyl-thiophen-2-yl)-
[1,2,4]oxadiazole (609 mg) as a yellow solid; LC-MS: tR = 1.19 min, [M+1] = 385.23.
b) To a solution of 3-(4-allyloxy-2-methoxy-phenyl)-5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazole (609 mg, 1.59 mmol) in acetone (15 mL) and water (1.5 mL), NMO (750 mg, 5.55 mmol) and OSO4 (0.1 mL, 2.5 % solution in tert.butanol, 8 nmol) is added. The mixture is stirred at rt for 48 h. The mixture is diluted with water (50 mL) and extracted with EA (100 mL, 2x 50 mL). The combined organic extracts are dried (Na2S04), filtered and evaporated. The residue is purified by CC on silica gel eluting with EA to give rac-3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-3-methoxy-phenoxy}-pro-pane-1,2-diol (480 mg) as a pale yellow solid; LC-MS: \R = 0.99 min, [M+1] = 419.19; ""H NMR (CD3OD): 6 7.96 (d, J = 8.8 Hz, 1H), 7.66 (s, 1H), 6.72-6.60 (m, 2H), 4.20-3.98 (m, 3H), 3.93 (s, 3H), 3.74-3.64 (m, 2H), 2.50-2.40 (m, 5H), 1.90 (hept, J = 7.0 Hz, 1H), 0.94 (d, J = 6.4 Hz, 6H).

To a solution of rac-3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yll-3-methoxy-phenoxy}-propane-1,2-diol (400 mg, 0.956 mmol) and DIPEA (257 mg, 1.99 mmol) in THF (40 mL), methanesulfonylchloride (125 mg, 1.095 mmol) dissolved in THF (10 mL) is added at -10°C. The reaction is allowed to warm to rt and is stirred for 15 h. Another portion of methanesulfonylchloride (30 mg, 0.258

mmol) in THF (2 mL) and DIPEA (128 mg, 0.993 mmol) is added and stirring is continued for 24 h. The mixture is evaporated and the residue is dissolved in 7 N NHs in MeOH (17 mL). The resulting solution is stirred at 70°C for 5 h before it is evaporated. The remaining residue is dissolved in DMF (15 mL) and glycolic acid (110 mg, 1.45 mmol), DIPEA (257 mg, 1.99 mmol) and TBTU (370 mg, 1.15 mmol) is added. The mixture is stirred at rt for 5 h, evaporated and the residue is purified on prep. TLC plates with EA to give 2-hydroxy-N-(2-hydroxy-3-{4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,2,4]oxadiazoi-3-yi]-3-methoxy-phenoxy}-propyl)-acetamide (135 mg) as colourless foam; LC-MS; tp = 0.95 min, [M+1] = 476.29; ^H NMR (De-acetone): 8 7.92 (d, J = 8.8 Hz, 1H), 7.71 (s, 1H), 7.57 (s br, 1H), 6.77 (d, J = 2.3 Hz, 1H), 6.71 (dd, J = 2.3, 8.8 Hz, 1H), 4.74-4.67 (m, 2H), 4.15-4.07 (m, 3H), 3.99 (d, J = 5.0 Hz, 2H), 3.93 (s, 3H), 3.66-3.57 (m, 1H), 3.48-3.39 (m, 1H), 2.52 (d, J = 7.0 Hz, 2H), 2.49 (s, 3H), 1.95 (hept, J = 7.0 Hz, 1H), 0.95 (d, J = 6.4 Hz, 6H).

rac-2-Hydroxy-N-(2-hydroxy-3-{4-[5-(4-isobutyl-3-methyl-thiophen-2-yl)-[1,2,4]oxadiazol-3-yl]-3-methoxy-phenoxy}-propyl)-acetamide is prepared from rac-

To a solution of Intermediate A1 (291 mg, 1.47 mmol) in DCM (10 mL), DIPEA (573 mg, 4.44 mmol) and TBTU (474 mg, 1.48 mmol) is added. The mixture is stirred for 15 min before 4-benzyloxy-3,5-dimethyl-benzoic acid hydrazide (396 mg, 1.47 mmol) is added. Stirring is continued at rt for 16 h. The mixture is diluted with diethyl ether, washed twice with 1 N aq. NaOH solution, once with 1 N aq. HCI solution, dried over Na2S04, and filtered. The solvent is evaporated to give 4-benzyloxy-3,5-dimethyl-benzoic acid N'-(4-isobutyl-5-methyl-thiophene-2-carbonyl)-hydrazide (371 mg) as a beige solid; LC-MS: tR = 1.07 min, [M+1] = 451.40. To a solution of this material in THF (5 mL), Burgess reagent (305 mg, 1.28 mmol) is added. The mixture is heated to 110°C for 5 min in a microwave oven. The mixture is cooled to rt, diluted with diethyl ether and washed with brine. The organic extract is dried over Na2S04, filtered and concentrated to give 2-(4-benzyloxy-3,5-dimethyl-phenyl)-5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazole (348 mg) as an orange oil; LC-MS: IR = 1.26 min, [M+1] = 433.40. This material is dissolved in THF:ethanol 1:1 (20 mL) and treated with a suspension of Pd/C (100 mg, 10% Pd) in ethanol. The mixture is stirred at rt for 3 h under 1 bar or H2. The catalyst is removed by filtration and the filtrate is evaporated to give 4-[5-(4-isobutyl-5-methyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenol (274 mg) as a grey solid, LC-MS: tR = 1.12 min, [M+l] = 343.25.

phase is about 7. The org. phase is separated and the aq. phase is extracted once
more with diethyl ether. The combined organic extracts are dried over MgS04,
filtered and concentrated. The crude product is purified by CC on silica gel eluting
with heptane-.EA 7:3 to give 3-(4-hydroxy-2-methoxy-phenyl)-1-(4-isobutyl-3,5-
dimethyl-thiophen-2-yl)-propenone (270 mg) as a yellow solid; LC-MS; tR = 1.07
min, [M+1] = 345.18. This material is hydrogenated as described in Example 7 to
give 3-(4-hydroxy-2-methoxy-phenyl)-1-(4-isobutyl-3,5-dimethyl-thiophen-2-yl)-
propan-1-one (203 mg) as an orange oil; LC-MS: tR = 1.07 min, [M+1] = 347.19.

(3-{4-[5-(4-lsobutyl-3,5-dimethyl-thiophen-2-yl)-[1,3,4]oxadiazol-2-yl]-2,6-dimethyl-phenyl}-propionylamino)-acetic acid is prepared from Intermediate D5 in following the procedures given in Examples 67 and 68; LC-MS: tp = 1.05 min, [M+1] =470.49; ^H NMR (De-DMSO, 100°C): 8 0.86 (d, J = 6.7 Hz, 6 H), 1.76 (hept, J = 6.7 Hz, 1H), 2.23-2.32 (m, 2 H), 2.35 (s, 6 H), 2.37 (s, 3 H), 2.38-2.42 (m, 2 H), 2.45 (s, 3 H), 2.82-2.91 (m, 2 H), 3.73 (s, 2 H), 7.59 (s, 2 H).
Example 113: GTPyS assay to determine EC50 values
GTPyS binding assays are performed in 96 well microtiter plates (Nunc, 442687) in a final volume of 200 pi, using membrane preparations of CHO cells expressing recombinant human SI PI receptor. Assay conditions are 20 mM Hepes (Fluka, 54461), 100 mM NaCI (Fluka, 71378), 5 mM MgCl2 (Fluka, 63064), 0.1% BSA (Calbiochem, 126609), 1 |JM GDP (Sigma, G-7127), 2.5% DMSO (Fluka, 41644), 50 pM 35S-GTPyS (Amersham Biosciences, SJ1320). The pH is 7.4. Test compounds are dissolved and diluted in 100% DMSO and pre-incubated at room temperature for 30 min in 150 μ of the above assay buffer, in the absence of 35S-GTPyS. After addition of 50 ΜI of 35S-GTPyS, the assay is incubated for 1 h at rt. The assay is terminated by transfer of the reaction mixture to a Multiscreen plate (Millipore, MAHFC1H60) using a cell harvester from Packard Biosciences, and the plates are washed with ice-cold 10 mM Na2HP04/NaH2P04 (70%/30%), dried, sealed at the bottom and, after addition of 25 \i\ MicroScint20 (Packard Biosciences, order# 6013621), sealed on the top. Membrane-bound 35S-GTPyS is measured with a TopCount from Packard Biosciences.
EC50 is the concentration of agonist inducing 50 % of the maximal specific 35S-GTPyS binding. Specific binding is determined by subtracting non-specific binding from maximal binding. Maximal binding is the amount of cpm bound to the

Multiscreen plate in the presence of 10 pM of S1P. Non-specific binding is the amount of binding in the absence of an agonist in the assay.
Table 1 shows the EC50 value of some compounds of the present invention. The EC50 values were determined according to the method described above:

Example 114: Assessment oi In vivo Efficacy
The efficacy of the compounds of Formula (I) is assessed by measuring the circulating lymphocytes after oral administration of 3 to 30 mg/kg of a compound of Formula (I) to normotensive male Wistar rats. The animals are housed in climate-controlled conditions with a 12 h-light/dark cycle, and have free access to normal rat chow and drinking water. Blood is collected before and 3, 6 and 24 h after drug administration. Full blood is subjected to hematology using Advia Hematology system (Bayer Diagnostics, Zurich, Switzerland).
All data are presented as mean ± SEM. Statistical analyses are performed by analysis of variance (ANOVA) using Statistica (StatSoft) and the Student-Newman-Keuls procedure for multiple comparisons. The null hypothesis is rejected when p < 0.05.

As an example, Table 2 shows the effect on lymphocyte counts 6 h after oral administration of 10 mg/kg of compounds of the present invention to normotensive male Wistar rats as compared to a group of animals treated with vehicle only.

Claims
1. A compound selected from the group consisting of thiophene compounds of the Formula (I),

wherein

wherein the asterisks indicate the bond that is linked to the thiophene group of Formula (I);
R2 represents hydrogen, methyl, or trifluoromethyl;
R2 represents n-propyl, isobutyl, or cyclopropylmethyl;
R' represents hydrogen, methyl, trifluoromethyl, ethyl, n-propyl, isopropyl, or isobutyl;
R* represents hydrogen, C1-4-alkyl, methoxy, or halogen;
R* represents hydrogen, C1-4-alkyl, C1-4-alkoxy, or halogen;

R® represents hydroxy-C1-4-alkyl, di-(hydroxy-C1-4-alkyl)-C1-4-alkyl, 2,3-dihydroxypropyl, -CH2-(CH2)n-NR2^R2^ -CH2-(CH2)n-NHCOR2^ -CH2-(CH2)n-NHS02R2^ -(CH2)k-(CHR2=)p-CHR2^-CONR2^R22 -(CH2)nCH(OH)-CH2-NR2^R2^ -(CH2)nCH(OH)-CH2-NHCOR2, -(CH2)nCH(OH)-CH2-NHS02R2^ -CO-NHR2\ hydroxy, hydroxy-C2-4-alkoxy, di-(hydroxy-Ciw,-alkyl)-C1-4-alkoxy, 1-glyceryl, 2,3-dihydroxypropoxy, 2-hydroxy-3-methoxy-propoxy, -OCH2-(CH2)m-NR2''R®^, -OCH2-(CH2)m-NHCOR2^ -OCH2-(CH2)m-NHS02R2^ -OCH2-CH(OH)-CH2-NR2^R2^ -OCH2-CH(0H)-CH2-NHC0R2, -OCH2-CH(OH)-CH2-NHS02R2^ -NR2^R2^ -NHCO-R2, or -S02NH-R2^
R®^ represents hydrogen, methyl, ethyl, 2-hydroxyethyl, 2-hydroxy-1-
hydroxymethyl-ethyl, 2,3-dihydroxy-propyl, 2-C1-4-alkoxyethyl, 3-hydroxypropyl, 3-
C1-4-alkoxypropyl, 2-amlnoethyl, 2-(C1-4-alkylamino)ethyl, 2-(di-(C1-4-
alkyl)amino)ethyl, carboxymethyl, C1-4-alkylcarboxymethyl, 2-carboxyethyl, or 2-(Ci-4-alkylcarboxy)ethyl;
R®^ represents hydrogen, or methyl;
R2^ represents methyl, ethyl, 2-hydroxyethyl, 2-methoxyethyl, methylamino, ethylamino, or dimethylamino;
R«* represents hydroxy methyl, aminomethyl, methylaminomethyl, dimethylaminomethyl, 2-amlnoethyl, or2-methylamino-ethyl;
R®^ represents hydrogen;
R®^ represents hydrogen or hydroxy; and
in case R2® represents hydroxy, R®* may in addition represent hydroxy;
m represents the integer 1 or 2;
n represents 0, 1, or 2;
k represents 0;
p represents 0 or 1; and

in case p represents 1, k may in addition represent 1; and R2 represents hydrogen, C1-4-alkyl, or lialogen; and salts tinereof.
2. A compound according to claim 1, wherein R2 represents hydroxy-C1-4-alkyl, di-
■ (hydroxy-C1-4-alkyl)-C1-4-all