NO330640B1 - Tetrahydroquinoline derivatives, pharmaceutical compositions comprising such, such compounds for use in therapy and the use thereof for the manufacture of fertility control drug - Google Patents

Description

The present invention relates to tetrahydroquinoline derivatives, pharmaceutical preparations comprising such, such compounds for use in therapy as well as the use of such for the production of medication for fertility control.

Gonadotropins serve important functions in a variety of bodily functions including metabolism, temperature regulation, and the reproductive process. Gonadotropins act on specific gonadal cell types to initiate ovarian and testicular differentiation and steroidogenesis. For example, the pituitary gonadotropin FSH (follicle-stimulating hormone) plays a central role in stimulating follicle development and maturation while LH (luteinizing hormone) induces ovulation (Sharp, R.M. Clin Endocrinol. 33:787-807, 1990; Dorrington and Armstrong, Recent . Prog. Horm. Res. 35:301-342, 1979). Currently, FSH is used clinically, in combination with LH or hCG, for ovarian stimulation, i.e. ovarian hyperstimulation for in vitro fertilization (IVF) and induction of ovulation in infertile anovulatory women (Insler, V., Int. J. Fertility 33 :85-97, 1988, Navot and Rosenwaks, J. Vitro Fert. Embryo Transfer 5:3-13, 1988), as well as for male hypogonadism and male infertility.

The gonadotropin FSH is released from the anterior pituitary under the influence of gonadotropin-releasing hormone and estrogens and from the placenta during pregnancy. In the woman, FSH acts on the ovaries and promotes the development of follicles and is the most important hormone for regulating the secretion of estrogens. In the male, FSH is responsible for the integrity of the seminiferous tubules and acts on Sertoli cells to support gametogenesis. Purified FSH is used clinically to treat infertility in women and for some types of spermatogenesis failure in men. Gonadotropins intended for therapeutic purposes can be isolated from human urine sources and are of low purity (Morse et al, Amer. J. Reproduct. Immunol. and Microbiology 17:143, 1988). Alternatively, they can be prepared as recombinant gonadotropins. Recombinant human FSH is available commercially and is used in assisted reproduction (Olijve et al. Mol. Hum. Reprod. 2:371, 1996; Devroey et al. Lancet 339:1170, 1992).

The effects of the FSH hormone are mediated by a specific plasma membrane receptor which is an element of the large family of G-protein coupled receptors. These receptors consist of a single polypeptide with seven transmembrane domains and are able to interact with the Gs protein, leading e.g. to the activation of adenylate cyclase.

The FSH receptor is a very specific target in the ovary-follicle growth process and is exclusively expressed in the ovary. Blocking this receptor or inhibiting the signaling that is normally induced after FSH-mediated receptor activation will disrupt follicle development and thus ovulation and fertility. FSH antagonists with low molecular weight could therefore form the basis for new contraceptives. Such FSH antagonists could give rise to reduced follicle development (no ovulation) with still sufficient residual estrogen production to avoid adverse effects on e.g. bone mass. On the other hand, compounds that stimulate FSH receptor activity may serve to mimic the gonadotropic effect of the natural ligand.

The production of low molecular weight hormone analogues which selectively have modulating activity on the FSH receptor is described below. The compounds can either be used as (partial) agonists or (partial) antagonists of the FSH receptor.

Thus, it has now been found that the following class of tetrahydroquinoline compounds of formula I or pharmaceutically acceptable salts thereof have fertility regulating activity:

where

R<1> and R<2> are Me;

R<3> is morpholinylcarbonylamino or R<5->(2-4C)alkyl;

R<4> is biphenyl or phenyl optionally substituted with halogen, (1-6C) alkoxy, (1-6C) alkyl, NO 2 , (di)(1-6C) alkylamino or CF 3 ;

R<5> is (di)(1-4C)alkylamino, morpholinyl, piperazinyl, pyridyl, piperidinyl optionally substituted with (1-6C)alkyl, azepanyl, aminocarbonyl,

(2-7C)alkenylaminocarbonyl, (1-6C)alkylamino, pyridyl-(1-4C)alkylaminocarbonyl, furyl-(1-4C)alkylaminocarbonyl, (1-6C)alkoxy-(1-6C)alkylaminocarbonyl, phenyl-(1 -4C)alkylaminocarbonyl, (1-6C)alkylaminocarbonyl, morpholinylcarbonyl, furyl-(1-4C)alkylamino, hydroxy-(1-6C)alkylamino, pyridyl-(1-6C)alkylamino, amino-(1-6C)alkylamino or morpholinylcarbonylamino.

The compounds above modulate FSH receptor function and can be used for the same clinical purposes as natural FSH if they act as agonists, with the advantage that they show altered stability properties and can be administered in different ways. If they block the FSH receptor, they can be used e.g. as a contraceptive.

Thus, the tetrahydroquinoline derivative according to the present invention can be used for fertility regulation such as the treatment of infertility and for contraception.

The following designations shall have the specified meanings given below unless otherwise defined in the requirements.

The term (1-4C)alkyl as used herein means a branched or unbranched alkyl group having 1-4 carbon atoms, and is methyl, ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl.

The term (2-4C)alkyl as used herein means a branched or unbranched alkyl group having 2-4 carbon atoms, and is ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl.

The term (1-6C)alkyl as used herein means a branched or unbranched alkyl group having 1-6 carbon atoms, for example methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl and hexyl. (1-5C)Alkyl groups are preferred, (1-4C)alkyl is the most preferred.

The term (di)(1-4C)alkylamino as used herein means an amino group, monosubstituted or disubstituted with alkyl groups each containing 1-4 carbon atoms and has the same meaning as previously defined.

The term (di)(1-4C)alkenylamino as used here means an amino group, monosubstituted or disubstituted with alkenyl groups each containing 2-4 carbon atoms such as allyl and 2-butenyl and has the same meaning as previously defined.

The term (3-8C)cycloalkyl as used herein means a cycloalkyl group having 3-8 carbon atoms, and is cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and cyclooctyl. (3-6C) cycloalkyl groups are preferred.

The term (2-6C)heterocycloalkyl as used herein means a heterocycloalkyl group having 2-6 carbon atoms, preferably 3-5 carbon atoms and which includes at least one heteroatom selected from N, O and/or S, which may be attached via a heteroatom if possible or a carbon atom. Preferred heteroatoms are N or O. The heterocycloalkyl group may be substituted with a methyl or ethyl group at a carbon atom or a heteroatom if possible. Most preferred heterocycloalkyl groups are piperidinyl, piperazinyl, morpholinyl, pyrrolidinyl and 1-methyl-2-piperidinyl.

The term (1-4C) alkoxy as used here means an alkoxy group having 1-4 carbon atoms, the alkyl group has the same meaning as previously defined.

(1-2C)Alkoxy groups are preferred.

The term (6C)aryl as used here means a phenyl group, which may optionally be substituted with one or more substituents selected from hydroxy, amino, iodo, bromine, chlorine, fluorine, nitro, trifluoromethyl, cyano, phenyl, (1-4C)alkyl , (1-4C)alkoxy or (1-4C)(di)alkylamino; The alkyl, alkoxy and (di)alkylamino groups have the same meaning as previously defined, for example phenyl, 3,5-dibromophenyl, 4-biphenyl, 3,5-dichlorophenyl, 3-bromo-6-methylamino-phenyl, 3-chloro-2, 6-dimethoxyphenyl and 3,5-dimethylphenyl.

The term (2-5C)heteroaryl as used herein means a substituted or unsubstituted aromatic group having 2-5 carbon atoms, which includes at least one heteroatom selected from N, O and/or S, such as imidazolyl, pyridyl, pyrimidyl, thienyl or furyl . The substituents on the heteroaryl group can be selected from the substituent group listed for the (6C)aryl group. The heteroaryl group may be attached via a carbon atom or a heteroatom, if possible. Preferred heteroaryl groups are thienyl, furyl and pyridyl.

The term (2-6C)heterocycloalkyl(1-4C)alkyl as used herein means a heterocycloalkyl group having 2-6 carbon atoms, connected to an alkyl group having 1-4 carbon atoms, the heterocycloalkyl group and the alkyl group having the same meaning as previously defined.

The term (2-6C)heterocycloalkylcarbonylamino as used herein means a heterocycloalkyl group having 2-6 carbon atoms, connected to the carbonyl group by a carbonylamino group, the heterocycloalkyl group having the same meaning as previously defined.

The term (2-6C)heterocycloalkylcarbonylamino(2-4C)alkyl as used herein means a heterocycloalkylcarbonylamino group in which the heterocycloalkyl group contains 2-6 carbon atoms, connected via the amino group to an alkyl group having 2-4 carbon atoms, the heterocycloalkylcarbonylamino group and the alkyl group have the same meaning as previously defined .

The term (di)(1-4C)alkylaminocarbonyl as used here means a (di)alkylamino group, the alkyl group(s) of which have 1-4 carbon atoms, connected via the amino group to a carbonyl group, the (di)alkylamino group has the same meaning as previously defined.

The term (3-8C)cycloalkylaminocarbonyl as used herein means a cycloalkyl group having 3-8 carbon atoms, connected to the amino group by an aminocarbonyl group, the cycloalkyl group having the same meaning as previously defined.

The term (di)(1-4C)alkylaminocarbonylamino as used herein means a (di)alkylamino group, the alkyl group(s) of which have 1-4 carbon atoms, connected via the amino group to the carbonyl group of a carbonylamino group, thus providing a urea functionality, (di) the alkylamino group has the same meaning as previously defined.

The term (di)(1-4C)alkylaminocarbonylamino(2-4C)alkyl as used herein means a (di)alkylaminocarbonylamino group, the alkyl group(s) thereof having 1-4 carbon atoms, connected via the amino group to an alkyl group having 2-4 carbon atoms , the (di)alkylaminocarbonylamino group and the alkyl group have the same meaning as previously defined.

The term (2-5C)heteroaryl(1-4C)alkyl as used here means a heteroaryl group having 2-5 carbon atoms connected to an alkyl group having 1-4 carbon atoms, the heteroaryl group and the alkyl group having the same meaning as previously defined.

The term (6C)aryl(1-4C)alkyl as used herein means phenyl group, optionally substituted with one or more substituents selected from the group of substituents listed for the (6C)aryl group, linked to an alkyl group having 1-4 carbon atoms, the aryl group and the alkyl group have same meaning as previously defined.

The term (6C)arylamino as used herein means phenyl group, optionally substituted with one or more substituents selected from the substituent group listed for the (6C)aryl group, connected to an amino group, the aryl group having the same meaning as previously defined.

The term (6C)aryl(1-4C)alkylamino as used herein means phenyl group, optionally substituted with one or more substituents selected from the group of substituents listed for the (6C)aryl group, connected to the alkyl group by an alkylamino group having 1-4 carbon atoms, the aryl group and the alkylamino group has the same meaning as previously defined.

The term (2-5C)heteroaryl(1-4C)alkylamino as used herein means a heteroaryl group having 2-5 carbon atoms, optionally substituted with one or

several substituents selected from the substituent group listed for the (6C)aryl group, connected to the alkyl group by an alkylamino group having 1-4 carbon atoms, the heteroaryl group and the alkylamino group have the same meaning as previously defined.

The term (1-4C)Alkoxy(2-4C)alkyl as used herein means an Alkoxy group having 1-4 carbon atoms, connected to an Alkyl group having 2-4 Carbon atoms, the Alkoxy group and the Alkyl group having the same meaning as previously defined

The term (di)[(1-4C)alkoxy(2-4C)alkyl]amino as used herein means an amino group, monosubstituted or disubstituted with (1-4C)alkyl(2-4C)alkyl groups. (1-4C)Alkoxy(2-4C)alkyl group is an alkoxy group that has 1-4 carbon atoms, connected to an alkyl group that has 2-4 carbon atoms and has the same meaning as previously defined.

The term (1-4C)alkylamino(2-4C)alkyl as used here means an alkylamino group having 1-4 carbon atoms, connected via the amino group to an alkyl group having 2-4 carbon atoms, the alkyl groups having the same meaning as previously defined.

The term (di)[(1-4C)alkylamino(2-4C)alkyl]amino as used herein means an amino group, monosubstituted or disubstituted with (1-4C)alkylamino(2-4C)alkyl groups. The (1-4C)alkylamino(2-4C)alkyl group is an alkylamino group that has 1-4 carbon atoms, connected via the amino group to an alkyl group that has 2-4 carbon atoms and has the same meaning as previously defined.

The term amino(2-4C)alkyl as used here means an aminoalkyl group having 2-4 carbon atoms, the alkyl group having the same meaning as previously defined.

The term (di)[amino(2-4C)alkyl]amino as used here means an amino group, monosubstituted or disubstituted with aminoalkyl groups having 2-4 carbon atoms and having the same meaning as previously defined.

The term hydroxy(2-4C)alkyl as used here means a hydroxyalkyl group having 2-4 carbon atoms, the alkyl group having the same meaning as previously defined.

The term (di)[hydroxy(2-4C)alkyl]amino as used here means an amino group, monosubstituted or disubstituted with hydroxyalkyl groups, which has 2-4 carbon atoms and which has the same meaning as previously defined.

The term R<5>(2-4C)alkyl as used here means an R<5> group attached to an alkyl group having 2-4 carbon atoms which has the same meaning as previously defined.

The term R<5>carbonyl-(1-4C)alkyl as used here means an R<5> group bound to the carbonyl group of a carbonylalkyl group, the alkyl group has 1-4 carbon atoms and has the same meaning as previously defined.

The term pharmaceutically acceptable salt represents those salts which are, within the scope of medical judgement, suitable for use in contact with the tissues of humans and lower animals without unnecessary toxicity, irritation, allergic response and are in accordance with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. They can be obtained during the final isolation and purification of the compounds according to the invention, or separately by reacting a free base function, if present, with a suitable mineral acid such as hydrochloric acid, phosphoric acid or sulfuric acid or with an organic acid such as for example ascorbic acid, citric acid, tartaric acid, lactic acid, maleic acid, malonic acid, fumaric acid, glycolic acid, succinic acid, propionic acid, acetic acid and methanesulfonic acid. If present, an acid function can be reacted with an organic or a mineral base, such as sodium hydroxide, potassium hydroxide or lithium hydroxide.

The invention thus deals with the compounds of formula I as defined here above.

In another embodiment, the invention deals with compounds of formula I, where R<4> is phenyl.

In yet another embodiment, the invention provides compounds of formula I where R<5> is (di)(1-4C)alkylamino furyl-(1-4C)alkylamino or pyridyl-(1-6C)alkylamino.

In yet another aspect of the invention, there are provided compounds of formula I wherein R 5 is (di)(1-4C)alkylamino.

Yet another aspect of the invention deals with compounds where all specific definitions of the groups R<1> to R<5> as defined above are combined in the compound of formula I.

Suitable methods for producing the compounds according to the invention are described below.

The compounds according to the present invention with formula 1-a can be prepared by starting with the well-documented Skraup reaction. Carrying out this reaction on N/-tert-butoxycarbonyl (/V-Boc) protected 1,4-phenylenediamine (II) gives 1,2-dihydroquinoline derivative III-a.

Related Skraup cyclocondensation reactions can be found in the literature: A. Knoe-venagel, Chem. Pray. 54:1726, 1921; R.L. Atkins and D.E. Bliss, J. Org. Chem. 43:1975, 1978; J.V. Johnson, B.S. Rauckman, D.P. Baccanari and B. Roth, J. Med. Chem. 32:1942, 1989; W.C. Lin, S.-T. Huang and S.-T. Lin, J. Chin. Chem. Soc.

43:497, 1996; J. P. Edwards, S.J. West, K.B. Marschke, D.E. Mais, M.M. Gottardis and T.K. Jones, J. Med. Chem. 41:303, 1998.

The above reaction is typically carried out at elevated temperature in acetone or mesityl oxide in the presence of iodine or protic acid such as hydrochloric acid, p-toluenesulfonic acid or aqueous hydrogen iodide. Alternatively, the compound of formula 11 1-a can be prepared by reacting compound II with acetone in the presence of MgSCv, 4-tert-butylcatechol and iodine (L.G. Hamann, R.l. Higuchi, L. Zhi, J.P. Edwards and X.-N. Wang, J .Med.Chem, 41:623, 1998). In yet another procedure, the reaction can be carried out in acetone using lanthanide triflates (eg, scandium triflate) as catalysts. In this case, the reaction can be run at room temperature or at elevated temperatures using conventional heating or microwave irradiation (M. E. Theoclitou and L. A. Robinson, Tetrahedron Lett. 43:3907, 2002).

The compound of formula III-b can be prepared from /V-Boc-1,4-phenylenediamine II by reaction with methyl vinyl ketone. Related cyclizations are described in United States Patent 2,686,182 (Badische Anilin- & Soda-Fabrik Aktiengesellschaft).

Subsequent 1-A/-acetylation of the compounds of formula III-a-b can be carried out using standard conditions. In a typical experiment, compounds of formula III-a-b are heated under reflux in acetic anhydride or reacted in a solvent such as dichloromethane, tetrahydrofuran, toluene or pyridine with acetyl chloride in the presence of a base such as /V,A/-diisopropylethylamine , triethylamine or sodium hydride, to give the 1-N-acetyl-4-methyl-1,2-dihydroquinoline derivatives of formula IV-a-b.

Standard cleavage of the Boe protecting group under conditions well known to those skilled in the art yields the 6-amino-1,2-dihydroquinoline derivatives of formula V-a-b. This reaction is typically carried out in dichloromethane in the presence of trifluoroacetic acid.

Subsequent 6-A/-acylation of the compounds of formula V-a-b can be carried out using standard conditions, to give compounds of the general formula Vl-a-b, where R<4> is as previously defined. For example, compounds of formula V-a-b are reacted in a solvent such as dichloromethane, tetrahydrofuran or toluene with an acyl halide (R<4->C(0)-Cl) or acid anhydride (R<4->C(0)-0-C (0)-R<4>) in the presence of a base such as A/,A/-diisopropylethylamine, triethylamine, pyridine or sodium hydride, to give 6-A/-acylated 4-methyl-1,2-dihydroquinoline derivatives of formula Vl-a-b. Alternatively, acylation of compounds of general formula V-a-b to give compounds of general formula Vl-a-b can also be achieved by reaction with an appropriate carboxylic acid (R<4->CO 2 H) in the presence of a coupling reagent such as O-(benzotriazole -1-yl)-A/,A/,A/',A/'-tetramethyluronium tetrafluoroborate (TBTU), O-(7-azabenzotriazol-1-yl)-A/,A/,A/',A/ -tetramethyluronium hexafluorophosphate (HATU) or bromotripyrrolidinophosphonium hexafluorophosphate (PyBrOP) and a tertiary base, e.g. /V,A/-diisopropylethylamine, in a solvent such as /V,/V-dimethylformamide or dichloromethane at ambient or elevated temperature.

Introduction of the required substituted phenyl group at position 4 of the dihydro-quinoline framework can be accomplished via Friedel-Crafts alkylation of anisole with the compounds of general structure Vl-a-b, to give compounds of the general formula Vll-a-b. This reaction is typically carried out at elevated temperatures either in anisole or in a suitable inert solvent such as heptane or hexane with anisole as the reagent, catalyzed by a Lewis acid (eg AICI 3 , AIBr 3 , FeCb or SnCU). Friedel-Crafts alkylations with 2,2,4-trimethyl,2-dihydroquinolines are described in the literature by B.A. Lugovik, L.G. Yudin and A.N. Kost, Dokl. Acad. Nauk SSSR, 170:340, 1966; B.A. Lugovik, L.G. Yudin, S.M. Vinogradova and A.N. Diet, Khim. Goat cycle. Soedin, 7:795, 1971.

Alternatively, A/-Boc-1,4-phenylenediamine II can be reacted with 2-(4-methoxyphenyl)-propene and formaldehyde in acetonitrile at ambient or elevated temperature, followed by 1 -/V-acetylation as described previously, to give the compound Vll-b where R<4>= O-ferf-Bu. Related cyclizations are described in literature: J.M. Mellorog G.D. Merriman, Tetrahedron, 51:6115, 1995. Cleavage of the Boe protecting group and subsequent acylation of the 6-amino function with an acyl halide (R<4->C(0)-Cl) as described above provides access to compounds of general structure Vll-b where R4 is as described previously.

Cleavage of the aromatic methyl ether in compounds of general formula Vlll-a-b gives 4-(4-hydroxyphenyl) substituted tetrahydroquinoline derivatives of general formula Vlll-a-b, and sets the stage for functionalization of the free OH group.

Demethylation reactions of aromatic methyl ethers are well known to those skilled in the art. In a typical experiment, demethylation is achieved after reaction of a compound of formula Vlll-a-b with BBr3 in an inert solvent such as dichloromethane at low to ambient temperature to give demethylated compounds of general formula Vlll-a-b. Alternatively, demethylation can be carried out after reaction of compounds of formula VIII-a-b with BF 3 Me 2 S complex at ambient temperature.

Selective O-alkylation of compounds of general formula VIII-a-b with functionalized alkyl halides of general formula IX-a leads to the formation of compounds of general formula I-a-b. Alkylation reactions of aromatic hydroxyl groups are well known in the art. Typically, a solution of a compound of general formula VIII-a-b in a suitable solvent such as 1,4-dioxane, tetrahydrofuran, dichloromethane, acetonitrile, acetone or N,N-dimethylformamide is treated with a base (e.g. A/,/V-diisopropylamine, triethylamine, K 2 CO 3 , Cs 2 CO 3 or NaOH) and the appropriate alkylating reagent of general formula IX-a, for example benzyl bromide, 3-(dimethylamino)-propyl chloride, 4-(2-chloroethyl)-morpholine, 2-picolyl chloride or 2-chloroacetamide. Alternatively, alkylation can be carried out by the known Mitsunobu-type alkylation. In that case, a solution of a compound of the general formula VIII-a-b in a suitable solvent such as 1,4-dioxane, tetrahydrofuran or dichloromethane is treated with (resin-bound) triphenylphosphine, diethyl- or di-tert-butyl-azodicarboxylate and a functionalized alcohol of general formula IX-b. In principle, both alkylation methods can be used for all R3 groups, but a suitable protecting group strategy may be necessary if R<3> contains a nucleophilic group such as a secondary amine or a hydroxyl group. Selection of a protecting group and deprotection conditions are trivial to those skilled in the art.

Another procedure for obtaining compounds according to the present invention starts with the alkylation of compounds of the general formula VIII-a-b with esters of the general formula X.

The alkylation reaction is typically carried out in the presence of a base such as N,N-d-isopropylethylamine or sodium hydride in a suitable solvent such as N,N-dimethylformamide or tetrahydrofuran at ambient or elevated temperature. The ester function in the resulting compounds of general formula Xl-a-b where A = Me or Et can then be selectively reduced under controlled conditions to give compounds of general formula Xlll-a-b using a suitable reducing agent such as lithium aluminum hydride at low temperature or sodium borohydride in an inert solvent such as tetrahydrofuran. The free hydroxyl group in compounds of general formula Xlll-a-b can then be reacted with 4-toluenesulfonyl chloride (Ts-CI) or methanesulfonyl chloride (Ms-CI) in an inert solvent such as 1,4-dioxane, A/,A/- dimethylformamide or THF in the presence of a suitable base such as triethylamine or pyridine to form a suitable leaving group (compounds of general formula XIV-a-b; LG = Ts or Ms, respectively). Nucleophilic substitution with a suitable nucleophile (amine or alkoxide) under conditions known to those skilled in the art then gives access to compounds of general formula 1-a-b where R<3>= R<5->

(2-4C)alkyl and R<5> are as defined previously.

Conversion of compounds of general formula Xl-a-b where A = fert-Bu to carboxylic acids of general formula XIl-a-b can be carried out by deprotection of the tert-butyl ester function. In a typical experiment, the terf-butyl ester of general formula Xl-a-b (A = ferf-Bu) is dissolved in dichloromethane and treated with a strong acid such as trifluoroacetic acid. The resulting carboxylic acids of general formula XIl-a-b can then be condensed with an appropriate alcohol or amine in the presence of a coupling agent such as O-(benzotriazol-1-yl)-A/,A/,A/',A/' -tetramethyluronium tetrafluoroborate (TBTU), 0-(7-azabenzotriazol-1-yl)-A/,A/,A/',A/-tetramethyluronium hexafluorophosphate (HATU) or bromotripyrrolidinophosphonium hexafluorophosphate (PyBrOP) and a tertiary base, f .ex. /V,A/-diisopropylethylamine, in a solvent such as A/,/V-dimethylformamide or dichloromethane at ambient or elevated temperature, to give compounds of general formula l-a-b where R<3>= R<5-> carbonyl(1-4C)alkyl and R<5> are as defined previously.

Some of the compounds according to the invention, which may be in the form of a free base, can be isolated from the reaction mixture in the form of a pharmaceutically acceptable salt. The pharmaceutically acceptable salts can also be obtained by treating the free base of formula I with an organic or inorganic acid such as hydrogen chloride, hydrogen bromide, hydrogen iodide, sulfuric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, maleic acid, malonic acid, methanesulfonic acid, fumaric acid, succinic acid, tartaric acid, citric acid, benzoic acid and ascorbic acid.

The compounds according to the present invention have at least one chiral carbon atom and can therefore be obtained as pure enantiomers or as a mixture of enantiomers or as a mixture of diastereomers. Methods for obtaining the pure enantiomers are well known in the art, e.g. crystallization of salts obtained from optically active acids and the racemic mixture or chromatography using chiral columns. For diastereomers, normal phase or reversed phase columns can be used.

The compounds according to the invention can form hydrates or solvates. It is known to those skilled in the art that charged compounds form hydrated species when lyophilized with water or form solvated species when concentrated in solution with a suitable organic solvent. The compounds of the present invention include the hydrates or solvates of the listed compounds.

For the selection of active compounds, testing at 10"<5>M must result in an activity of more than 20% of the maximum activity when FSH is used as a reference. Another criterion could be the EC50 value which must be < 10"<5> M, preferably < 10"7 M.

Those skilled in the art will appreciate that desirable EC50 values are dependent on the compound tested. For example, a compound with an EC50 less than 10"<5>M is generally considered a candidate for drug selection. Preferably, this value is lower than 10"<7>M. However, a compound that has a higher EC50 but is selective for that particular receptor may be an even better candidate.

Methods for determining receptor binding, as well as in vitro and in vivo assays for determining biological activity of gonadotropins are well known. Generally, the expressed receptor is contacted with the compound to be tested and binding or stimulation or inhibition of a functional response is measured.

To measure a functional response, isolated DNA encoding the FSH receptor gene, preferably the human receptor, is expressed in suitable host cells. Such a cell could be a Chinese hamster ovary cell, but other cells are also suitable. Preferably, the cells are of mammalian origin (Jia et al, Mol. Endokrin., 5:759-776, 1991).

Methods for constructing recombinant FSH expressing cell lines are well known in the art (Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, latest edition). Receptor expression is achieved by expression of the DNA that codes for the desired protein. Techniques for site-directed mutagenesis, ligation of additional sequences, PCR and construction of suitable expression systems are all, by now, well known in the art. Portions, or all, of the DNA encoding the desired protein can be synthetically constructed using standard solid phase techniques, preferably to include restriction sites for easy ligation. Suitable control elements for transcription and translation of the included coding sequence can be provided to the DNA coding sequences. As is well known, expression systems are now available that are compatible with a variety of hosts, including prokaryotic hosts such as bacteria and eukaryotic hosts such as yeast, plant cells, insect cells, mammalian cells, avian cells and the like.

Cells expressing the receptor are then contacted with the test compound to observe binding or stimulation or inhibition of a functional response.

Alternatively, isolated cell membranes containing the expressed receptor can be used to measure compound binding.

For measurement of binding, radioactively labeled or fluorescently labeled compounds can be used. Competitive binding attempts can also be performed.

Another experiment involves screening for FSH receptor-agonist compounds by determining stimulation of receptor-mediated cAMP accumulation. Thus, such a method involves expression of the receptor on the cell surface of a host cell and exposure of the cell to the test compound. The amount of cAMP is then measured. The level of cAMP can be decreased or increased, depending on the inhibitory or stimulatory effect of the test compound after binding to the receptor.

Screening for FSH receptor antagonists involves incubating FSH receptor-expressing cells with a concentration range of the test compound in the presence of a fixed, submaximal effective, FSH concentration (ie, an FSH concentration that induces approximately 80% of the maximal stimulation of cAMP accumulation in the absence of test compound ). From the concentration-effect curves, the IC50 value and the percentage of inhibition of FSH-induced cAMP accumulation can be determined for each of the test compounds. Human recombinant FSH can be used as a reference compound. Alternatively, competitive trials can also be carried out.

In addition to direct measurement of e.g. cAMP levels in the exposed cell, cell lines can be used which, in addition to transfection with a receptor that codes for DNA, are also transfected with another DNA that codes for a reporter gene, the expression of which responds to the level of cAMP. Such reporter genes could be cAMP inducible or could be engineered in such a way that they are linked to new cAMP responsive elements. In general, reporter gene expression could be controlled by any response element that responds to changing levels of cAMP. Suitable reporter genes are e.g. the genes that code for P-galactosidase, alkaline phosphatase, firefly luciferase and green fluorescence protein. The principles of such transactivation experiments are well known in the field and are described e.g. in Stratowa, Ch., Himmler, A. and Czernilofsky, A.P., (1995) Curr.Opin.Biotechnol. 6:574.

The present invention also relates to a pharmaceutical preparation comprising a tetrahydroquinoline derivative or pharmaceutically acceptable salts thereof which have the general formula I in admixture with pharmaceutically acceptable additives and possibly other therapeutic agents. The additives must be "acceptable" in the sense of being compatible with the other components of the preparation and not harmful to the recipients thereof.

Preparations include e.g. they are suitable for oral, sublingual, subcutaneous, intravenous, intramuscular, local or rectal administration, all in unit dosage forms for administration.

For oral administration, the active ingredient can be presented as separate units, such as tablets, capsules, powders, granules, solutions and suspensions.

For parenteral administration, the pharmaceutical preparation according to the invention can be presented in unit dose or multidose containers, e.g. injection fluids in predetermined quantities, for example in sealed vials and ampoules and can also be stored in a freeze-dried (lyophilized) state that only requires the addition of a sterile liquid carrier, e.g. water, before use.

Mixed with such pharmaceutically acceptable additives, e.g. as described in the standard reference, Gennaro, A.R. et al., Remington: The Science and Practice of Pharmacy (20th Ed., Lippincott Williams & Wilkins, 2000, see especially Part 5: Pharmaceutical Manufacturing), the active agent may be compressed into fixed dosage units, such as pills, tablets, or processed into capsules or suppositories. By means of pharmaceutically acceptable liquids, the active agent can be used as a fluid composition, e.g. as an injection preparation, in the form of a solution, suspension, emulsion or as a spray, e.g. a nasal spray.

In order to produce fixed dose units, the use of conventional additives such as fillers, coloring agents and polymeric binders is envisaged. In general, any pharmaceutically acceptable additive which does not interfere with the function of the active compounds may be used. Suitable carriers with which the active agent according to the invention can be administered as solid preparations including lactose, starch, cellulose derivatives or mixtures thereof, used in suitable amounts. For parenteral administration, aqueous suspensions, isotonic saline solutions, and sterile injectable solutions may be used, containing pharmaceutically acceptable dispersing agents and/or wetting agents, such as propylene glycol or butylene glycol.

The above tetrahydroquinoline derivatives can also be administered in the form of implantable pharmaceutical devices, consisting of a core of active material, encapsulated by a release rate regulating membrane. Such implants are to be used subcutaneously or locally and will release the active ingredient at an approximately constant rate over relatively long periods of time, for example from weeks to years. Methods for producing implantable pharmaceutical devices as such are known in the field, for example as described in European Patent 0.303.306 (AKZO Nobel N.V.).

The exact dose and regimen of administration of the active ingredient or a pharmaceutical preparation thereof will necessarily depend on the therapeutic effect to be achieved (treatment of infertility; contraception) and may vary with the particular compound, route of administration and age and condition - that of the individual patient to whom the drug is to be administered.

In general, parenteral administration requires lower doses than other administration methods that are more dependent on absorption. However, a dose for humans preferably contains 0.0001-25 mg per kg body weight. The desired dose may be presented as a single dose or as multiple subdoses administered at appropriate intervals throughout the day, or, in the case of female recipients, as doses to be administered at appropriate daily intervals throughout the menstrual cycle. The dose as well as the administration regimen may differ between a female and a male recipient.

Thus, the compounds according to the invention can be used in therapy.

A further aspect according to the invention relates to the use of a tetrahydroquinoline derivative compound having the general formula I for the preparation of a drug to be used for fertility control.

In yet another aspect, the invention relates to the use of a tetrahydroquinoline derivative compound having the general formula I for the preparation of a medicament to be used for the treatment of infertility.

In yet another aspect, the invention relates to the use of a tetrahydroquinoline derivative compound having the general formula I for the preparation of a medicament to be used to prevent fertility.

The invention is illustrated by the following examples.

Examples

General comments

The following abbreviations are used in the examples: DMA = N,N-dimethylaniline, DIPEA = N,N-diisopropylethylamine; TFA = trifluoroacetic acid, DtBAD = di-tert-butyl azodicarboxylate; TBTU = O-benzotriazol-1-yl-N,N,N,N-tetramethyluronium tetrafluoroborate; HATU = O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate; Fmoc = 9-fluorenylmethoxycarbonyl; Fmoc-Cl = 9-fluorenyl-methoxycarbonyl chloride; DMF = N,N-dimethylformamide; Boe = tert -butoxycarbonyl; THF = tetrahydrofuran.

The names of the final products described in the examples have been formed using the Beilstein Autonom program (version: 2.02.119).

Unless otherwise indicated, all final products of the examples below are lyophilized from water/1,4-dioxane mixtures or water/acetonitrile mixtures. If the compound was prepared as an HCl or TFA salt, the respective acids were added in appropriate amounts to the solvent mixture prior to lyophilization.

The following analytical HPLC methods are used for determination of retention times: Method 1: Column: 5 µm Luna C-18(2) 150x4.6 mm; flow: 1 ml/min; detection: 210 nm; column temperature: 40°C; solvent A: CH3CN/H2O = 1/9 (v/v); solvent B: CH3CN; solvent C: 0.1 M aqueous trifluoroacetic acid; gradient: solvent A/B/C = 65/30/5 to 10/85/5 (vol/vol/v) for 30.00 min, then constant for another 10.00 min at A/B/C = 10/ 85/5 (volume/volume/v).

Method 2: Identical to Method 1, except for the gradient used: Gradient: solvent A/B/C = 75/20/5 to 15/80/5 (vol/vol/v) for 30.00 min, then constant for another 10.00 min at A/B/C = 15/80/5 (volume/volume/v).

Method 3: Column: 3 um Luna C-18(2) 100x2 mm; flow: 0.25 ml/min; detection: 210 nm; column temperature: 40°C; solvent A: H 2 O; solvent B: CH3CN; solvent C: 50 mM phosphate buffer, pH 2.1; gradient: solvent A/B/C = 70/20/10 to 10/80/10 (v/v/v) for 20.00 min, then constant for another 10.00 min at A/B/C = 10/ 80/10 (volume/volume/v).

Method 4: Identical to Method 3, except for the gradient used: Gradient: solvent A/B/C = 65/30/5 to 10/85/5 (vol/vol/v) for 20.00 min, then constant for another 10.00 min at A/B/C = 10/85/5 (volume/volume/v).

Method 5: Identical to Method 3, except for the gradient used: Gradient: solvent A/B = 75/25 to 0/100 (v/v) for 20.00 min, then constant for another 10.00 min at A/ B/C = 0/100 (volume/volume).

Method 6: Identical to Method 1, except for the gradient used: Gradient: solvent A/B/C = 35/60/5 to 10/85/5 (vol/vol/v) for 30.00 min, then constant for another 10.00 min at A/B/C = 10/85/5 (volume/volume/v).

The following methods are used for preparative HPLC purifications:

Method A: Column = Luna C-18. Gradient: 0.1% trifluoroacetic acid in H2O/CH3CN (9/1, v/v)/CH3CN = 100/0 to 0/100 (v/v) for 30-45 min, depending on ease of separation. Detection: 210 nm. The appropriate fractions were collected and concentrated (partially) in vacuo.

Method B: Column = Luna C-18. Gradient: H20/CH3CN (9/1, v/v)/CH3CN = 80/20 to 0/100 (v/v) for 30-45 min, depending on ease of separation. Detection: 210 nm.

Example 1

Biphenyl- 4- carboxylic acid { 1- acetyl- 4-[ 4-( 2- dimethylamino- ethoxy)- phenyl- 2, 2, 4-trimethyl- 1, 2, 3, 4- tetrahydro- quinolin- 6- yl)- amide (a) . ( 2, 2, 4- trimethvl- 1, 2- dihydroquinoline- 6- vl)- carbamine tert- butyl ester A mixture of /V-Boc-1,4-phenylenediamine (75 g), MgSCv (216 g), 4 -terf-butylcatechol (1.8 g) and iodine (4.7 g) in anhydrous acetone (600 ml) were heated under reflux for 20 h. MgSCv was removed by filtration and the filtrate was concentrated in vacuo. The residue was chromatographed on a short plug of silica gel using heptane/ethyl acetate = 8/2 (v/v) as the eluent to give the product as a brown oil.

Yield: 41 g.

(b) . ( 1- acetvl- 2, 2, 4- trimethylvl- 1, 2- dihydroquinoline- 6- vl)- carbamine tert- butyl ester A solution of the compound described in example 1a (41 g) in pyridine (200 ml) and CH 2 Cl 2 ( 200 ml) was cooled to 0°C. Acetyl chloride (21 mL) in CH 2 Cl 2 (50 mL) was added dropwise. After complete addition, the mixture was stirred for 3 hours at room temperature. Ethyl acetate (2 L) and H 2 O (2 L) were added and the organic layer was separated, dried and concentrated in vacuo. The title compound was obtained by crystallization from ethyl acetate.

Yield: 23 g.

(c) . 1 - acetyl- 6- amino- 2, 2, 4- trimethyl- 1, 2- dihydroquinoline

The compound described in Example 1b (15 g) was stirred in a mixture of CH 2 Cl 2 and TFA (9/1 (v/v), 300 ml) for 2 hours. The reaction mixture was cooled to 0°C and pH adjusted to pH 7 using a 2 M aqueous NaOH solution. The organic layer was separated, washed with brine, dried and concentrated in vacuo to give the crude product which was used without further purification in the next step.

Yield: 10.4 g

(d) . Biphenyl-4-carboxylic acid (1-acetyl-2,2,4-trimethyl-1,2-dihydroquinolin-6-yl)-amide To a solution of the compound described in Example 1c (10 g) and DIPEA (40 ml) in CH 2 Cl 2 (100 mL), 4-biphenylcarbonyl chloride (9.8 g) was added and the resulting

the mixture was stirred for 18 hours at room temperature. Water was added, the organic layer was separated, dried and concentrated in vacuo. The product was crystallized from ethyl acetate.

Yield: 15 g

(e) Biphenyl-4-carboxylic acid [1-acetyl-4-(4-methoxyphenyl)-2,2,4-trimethyl-1,2,3,4-tetrahydroquinoline-6-van-amide

With stirring, aluminum trichloride (9.7 g) was added to a mixture of the compound described in Example 1d (10.0 g) and anhydrous anisole (50 ml) and the resulting mixture was stirred at 35°C for 18 hours. After this time, water was added at 0°C and the resulting mixture was extracted with ethyl acetate. The organic layer was separated, dried and partially concentrated in vacuo and the mixture was stored at 0°C for 18 hours. The precipitate formed was collected by filtration and dried in vacuo to give the title compound.

Yield: 7.9 g.

(f) . Biphenyl- 4- carboxylic acid [ 1 - acetyl- 4-( 4- hydroxyphenyl)- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydroquinoline- 6- n- amide

To a solution of the compound described in Example 1e (7.9 g) in CH 2 Cl 2 (200 ml) at 0°C was added a solution of boron tribromide (5 ml) in CH 2 Cl 2 (50 ml) and the mixture was kept for 4 hours at 0°C. Water (about 500 ml) was carefully added and the resulting mixture was vigorously stirred. The organic layer was separated, dried and concentrated in vacuo. Crystallization from ethyl acetate gave the title compound. Yield: 6.1 g.

(g) Biphenyl-4-carboxylic acid {1-acetyl-4- r4-(2-dimethylamino-ethoxy)-phenyl-2,2,4-trimethyl-1,2,3,4-tetrahydroquinoline-6-yl )- amide

General procedure A: To a solution of the compound described in Example 1f (70 mg) in DMF (2 ml) was added Cs 2 CO 3 (200 mg) and 2-dimethylaminoethyl chloride hydrochloride (17 mg). The resulting mixture was stirred overnight, after which water and ethyl acetate were added. The organic layer was separated, dried and concentrated in vacuo. The product was purified by preparative HPLC (Method A) and lyophilized from a mixture of CH 3 CN and water containing TFA to give the corresponding TFA salt.

Yield: 18 mg (TFA salt); MS-ESI: [M+H]<+>= 576.6; HPLC: Rt = 14.96 min (method 3).

Example 2

Biphenyl- 4- carboxylic acid { 1- acetyl- 4- r4-( 2- dimethylamino- propoxy)- phenyl- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro- quinolin- 6- yl)- amide

According to General Procedure A, the compound described in Example 1f (70 mg) was alkylated with 3-dimethylaminopropyl chloride hydrochloride (19 mg) and CS 2 CO 3 (200 mg) in DMF (2 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 58 mg (TFA salt); MS-ESI: [M+H]<+>= 590.4; HPLC: Rt = 15.36 min (method 3).

Example 3

Biphenyl- 4- carboxylic acid { 1- acetyl- 2, 2, 4- trimethyl- 4- r4-( 3- morpholine- 4- yl- propoxy)- phenyl- 1, 2, 3, 4- tetrahydro- quinoline- 6- vl)- amide

According to General Procedure A, the compound described in Example 1f (70 mg) was alkylated with 3-morpholinopropyl chloride (26 mg) and CS 2 CO 3 (200 mg) in DMF (2 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt. Yield: 56 mg (TFA salt); MS-ESI: [M+H]<+>= 631.6; HPLC: Rt = 15.40 min (method 3).

Example 4

Biphenyl- 4- carboxylic acid { 1- acetyl- 2, 2, 4- trimethyl- 4- r4-( pvridin- 2- vlmethoxy) phenyl-1, 2, 3, 4- tetrahydro- quinolin- 6- vl)- amide

According to General Procedure A, the compound described in Example 1f (100 mg) was alkylated with 2-picolyl chloride hydrochloride (33 mg) and Cs 2 CO 3 (325 mg) in DMF (5 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt. Yield: 60 mg (TFA salt); MS-ESI: [M+H]<+>= 596.4; HPLC: Rt = 19.75 min (method 2).

Example 5

Biphenyl- 4- carboxylic acid { 1- acetyl- 2, 2, 4- trimethyl- 4- r4-( 1- methyl- piperidine- 3- vylmethoxy)- phenyl- 1, 2, 3, 4- tetrahydroquinoline- 6- vl}- amide

According to General Procedure A, the compound described in Example 1f (100 mg) was alkylated with 3-chloromethyl-1-methylpiperidine hydrochloride (33 mg) and CS 2 CO 3 (325 mg) in DMF (5 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 60 mg (TFA salt); MS-ESI: [M+H]<+>= 615.4; HPLC: Rt = 16.70 min (method 2).

Example 6

Biphenyl- 4- carboxylic acid { 1- acetyl- 4-[ 4-( 2- diethylamino- ethoxy)- phenyl- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro- quinolin- 6- yl)- amide

According to general procedure A, the compound described in Example 1f (100 mg) was alkylated with 2-diethylamino-ethyl chloride hydrochloride (35 mg) and CS 2 CO 3 (325 mg)

in DMF (5 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 67 mg (TFA salt); MS-ESI: [M+H]<+>= 604.4; HPLC: Rt = 16.38 min (method 2).

Example 7

Biphenyl-4-carboxylic acid {1-acetyl-2,2,4-trimethyl-4- r4-(pyridin-4-ylmethoxy)-phenyl-1,2,3,4-tetrahydro-quinolin-6-yl)-amide

According to General Procedure A, the compound described in Example 1f (100 mg) was alkylated with 4-picolyl chloride hydrochloride (33 mg) and Cs 2 CO 3 (325 mg) in DMF (5 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt. Yield: 61 mg (TFA salt); MS-ESI: [M+H]<+>= 596.4; HPLC: Rt = 16.64 min (method 2).

Example 8

Morpholine- 4- carboxylic acid r3-( 4-{ 1- acetyl- 6- r( biphenyl- 4- carboxylic acid)- aminol- 2, 2, 4-trimethyl- 1, 2, 3, 4- tetrahydroquinoline- 4- vl)-phenoxv)-propvll-amide According to general procedure A, the compound described in Example 1f (100 mg) was alkylated with morpholine-4-carboxylic acid (3-chloropropyl)amide (53 mg) and CS 2 CO 3 (325 mg) in DMF (5 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water.

Yield: 95 mg; MS-ESI: [M+H]<+>= 675.6; HPLC: Rt = 18.24 min (method 3).

Example 9

Biphenyl- 4- carboxylic acid 11 - acetyl- 4-[ 4-( 2- azepan- 1 - v- ethoxy)- phenyl- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro- quinoline- 6- vl)- amide

According to General Procedure A, the compound described in Example 1f (100 mg) was alkylated with 2-(hexamethyleneimino)ethyl chloride hydrochloride (42 mg) and CS 2 CO 3 (325 mg) in DMF (5 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 60 mg (TFA salt); MS-ESI: [M+H]<+>= 630.6; HPLC: Rt = 17.25 min (method 2).

Example 10

Biphenyl-4-carboxylic acid {1-acetyl-2,2,4-trimethyl-4-[4-(pyridin-3-ylmethoxy)-phenyl-1. 2, 3, 4-tetrahydroquinolin-6-yl)-amide

According to General Procedure A, the compound described in Example 1f (1.0 g) was alkylated with 3-picoloyl chloride hydrochloride (488 mg) and Cs 2 CO 3 (3.2 mg) in DMF (10 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt. Yield: 884 mg (TFA salt); MS-ESI: [M+H]<+>= 596.4; HPLC: Rt = 16.55 min (method 3).

Example 11

Biphenyl- 4- carboxylic acid H-acetyl- 4-( 4- carbamolmethox- phenyl)- 2, 2, 4- trimethyl-1. 2, 3, 4-tetrahydroquinoline-6-vn-amide

According to general procedure A, the compound described in Example 1f (100 mg) was alkylated with 2-chloroacetamide (24 mg) and Cs 2 CO 3 (325 mg) in DMF (5 mL). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 40 mg; MS-ESI: [M+H]<+>= 562.6; HPLC: Rt = 21.63 min (method 2).

Example 12

Biphenyl- 4- carboxylic acid ri-acetyl- 4-( 4- allylcarbamoylmethoxy- phenyl)- 2, 2, 4- trimethyl-1. 2, 3, 4-tetrahydroquinoline-6-vn-amide

(a) . ( 4-{ 1- acetvl- 6- r( biphenvl- 4- carbonvl) amino1- 2, 2, 4- trimethvl- 1, 2, 3, 4- tetrahydro-quinoline- 4- vl) phenoxv) acetic acid- tetra- butyl ester

A mixture of the compound described in Example 1f (2.58 g), tert-butyl bromoacetate (826 µl). K 2 CO 3 (2.8 g) and acetone (100 ml) were stirred for 18 h at 50°C. The solids were removed by filtration and the filtrate was concentrated in vacuo to give the product which was used without further purification in the next step.

Yield: 3.2 g

(b) . ( 4-{ 1- acetvl- 6- r( biphenvl- 4- carbonvl) amino1- 2. 2, 4- trimethvl- 1, 2, 3, 4- tetrahydroquinoline- 4- vl) phenoxv) acetic acid

The compound described in Example 12a (3.2 g) was stirred in a mixture of CH 2 Cl 2 and TFA (9/1 (v/v), 100 ml) for 3 hours. Toluene (100 mL) was added and the mixture was concentrated in vacuo to give the crude product, which was used without further purification.

Yield: 3.3 g

(c) . Biphenyl- 4- carboxylic acid ri- acetyl- 4-( 4- allylcarbamoyl methoxy- phenyl)- 2, 2, 4-trimethyl- 1, 2, 3, 4- tetrahydro- quinolin- 6- van- amide

General Procedure B: To a solution of the compound described in Example 12b (82 mg), allylamine (37 mg) and DIPEA (226 µl) in CH 2 Cl 2 (5 mL) was added TBTU (84 mg) at room temperature. If the reaction did not reach completion after 18 h, more TBTU and DIPEA were added. After completion of the reaction, water was added, the organic layer was separated, washed with brine, dried and concentrated in vacuo. The title compound was purified by preparative HPLC (Method A).

Yield: 48 mg; MS-ESI: [M+H]<+>= 602.4; HPLC: Rt = 18.19 min (method 4).

Example 13

Biphenyl-4-carboxylic acid {1-acetyl-4- r4-(isopropylcarbamoylmethoxy)-phenyl-2,2,4-trimethyl-1,2,3,4-tetrahydro- quinolin-6-yl)-amide

According to General Procedure B, the compound described in Example 12b (82 mg) was treated with isopropylamine (38 mg), DIPEA (226 µl) and TBTU (84 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 45 mg; MS-ESI: [M+H]<+>= 604.6; HPLC: Rt = 18.63 min (method 4).

Example 14

Biphenyl- 4- carboxylic acid H- acetyl- 4-( 4- diethylcarbamoylmethoxy- phenyl)- 2, 2, 4-trimethyl- 1, 2, 3, 4- tetrahydro- quinolin- 6- van- amide

According to General Procedure B, the compound described in Example 12b (82 mg) was treated with diethylamine hydrochloride (47 mg), DIPEA (226 µl) and TBTU (84 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 51 mg; MS-ESI: [M+H]<+>= 618.4; HPLC: Rt = 19.09 min (method 4).

Example 15

Biphenyl- 4- carboxylic acid riacetyl- 2, 2, 4- trimethyl- 4-( 4-{ r( pyridine- 4- dimethyl)-carbamoyl- methoxyphenyl)- 1, 2, 3, 4- tetrahydroquinoline- 6 -vll-amide According to general procedure B, the compound described in Example 12b (82 mg) was treated with 4-picolylamine (70 mg), DIPEA (226 µl) and TBTU (84 mg) in CH 2 Cl 2 (5 ml). The title compound was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 52 mg (TFA salt); MS-ESI: [M+H]<+>= 653.6; HPLC: Rt = 11.31 min (method 4).

Example 16

Biphenyl- 4- carboxylic acid ri- acetyl- 4-( 4-{[( furan- 2- methylethyl)- carbamov- methoxy)-phenyl)- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro- quinoline-6-vn-amide

According to General Procedure B, the compound described in Example 12b (82 mg) was treated with 2-furfurylamine (63 mg), DIPEA (226 µl) and TBTU (84 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 50 mg; MS-ESI: [M+H]<+>= 642.6; HPLC: Rt = 21.31 min (method 3).

Example 17

Biphenyl-4-carboxylic acid (1-acetyl-4-{4- r(2-methoxysulfetiylcarbamol)-methoxys1phenyl)-2. 2. 4- trimethyl- 1. 2. 3. 4- tetrahydro- quinolin- 6- vl)- amide

According to general procedure B, the compound described in Example 12b (82 mg) was treated with 2-methoxyethylamine (49 mg), DIPEA (226 µl) and TBTU (84 mg)

in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 34 mg; MS-ESI: [M+H]<+>= 620.4; HPLC: Rt = 19.70 min (method 3).

Example 18

Biphenyl- 4- carboxylic acid {1- acetyl-4- r4-( benzylcarbamoylmethoxy)- phenyl- 2, 2, 4-trimethyl- 1, 2, 3, 4- tetrahydro- quinolin- 6- yl)- amide

According to General Procedure B, the compound described in Example 12b (82 mg) was treated with benzylamine (49 mg), DIPEA (226 µl) and TBTU (84 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 53 mg; MS-ESI: [M+H]<+>= 652.6; HPLC: Rt = 22.26 min (method 3).

Example 19

Biphenyl- 4- carboxylic acid ( 1- acetyl- 4-{ 4-[( 2- dimethylamino- ethylcarbamoyl)- methoxy- phenyl)- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro- quinoline- 6 - vl)- amide

According to General Procedure B, the compound described in Example 12b (82 mg) was treated with A/,A/-dimethylethylenediamine (49 mg), DIPEA (226 µl) and TBTU (84 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Lyophilization from a mixture of aqueous HCl and 1,4-dioxane afforded the title compound as an HCl salt.

Yield: 11 mg (HCl salt); MS-ESI: [M+H]<+>= 633.4; HPLC: Rt = 13.74 min (method 3).

Example 20

Biphenyl- 4- carboxylic acid H- acetyl- 2, 2, 4- trimethyl- 4-( 4- methylcarbamov methoxyphenvl)-1, 2, 3, 4- tetrahydro- quinolin- 6- vn- amide

According to General Procedure B, the compound described in Example 12b (82 mg) was treated with methylamine hydrochloride (20 mg), DIPEA (226 µl) and TBTU (84 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 35 mg; MS-ESI: [M+H]<+>= 576.4; HPLC: Rt = 19.25 min (method 3).

Example 21

Biphenyl- 4- carboxylic acid { 1- acetyl- 2, 2, 4- trimethyl- 4-[ 4-( 2- morpholin- 4- yl- 2- oxo-ethoxy)- phenyl- 1, 2, 3, 4- tetrahydro - quinoline-6-vl)-amide

According to General Procedure B, the compound described in Example 12b (110 mg) was treated with morpholine (74 mg), DIPEA (296 µl) and TBTU (109 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A).

Yield: 85 mg; MS-ESI: [M+H]<+>= 632.4; HPLC: Rt = 12.48 min (method 3).

Example 22

A/-{ 1- acetvl- 2, 2, 4- trimethyl- 4- r4-( 3- morpholine- 4- vl- propoxy)- phenvn- 1, 2, 3, 4- tetrahydro-quinoline- 6- vl) - 5-bromo-2-methylamino-benzamide

(a). ( 1- acetvl- 2, 2, 4- trimethylvl- 1, 2- dihydro- quinoline- 6- vl)- carbamic acid 9- fluorene-vlmetvl ester

To a solution of the compound described in Example 1c (17 g) and DIPEA (40 mL) in CH 2 Cl 2 (100 mL), FmocCl (25 g) was added and the resulting mixture was stirred for 18 h at room temperature. Ethyl acetate (ca. 200 ml) and water (150 ml) were added, the organic layer was separated, dried and concentrated in vacuo. The title compound was purified by chromatography on silica gel using CH 2 Cl 2 as the eluent.

Yield: 16.6 g

(b) f1 - acetvl- 4-( 4- methoxysulfenvl)- 2, 2, 4- trimethvl- 1, 2, 3, 4- tetrahydroquinoline- 6- vll-carbamic acid 9- fluorenyl methyl ester

With stirring, aluminum trichloride (24.2 g) was added to a mixture of the compound described in Example 22a (16.5 g) and anhydrous anisole (150 ml) and the resulting mixture was stirred at 35°C for 18 hours. After this time, water was added at 0°C and the resulting mixture was extracted with ethyl acetate. The organic layer was separated, dried and partially concentrated in vacuo and the mixture was stored at 0°C for 18 hours. The precipitate formed was collected by filtration and dried in vacuo to give the title compound.

Yield: 10.1 g.

(c) ri-acetyl- 4-( 4- hydroxyphenyl)- 2. 2. 4- trimethyl- 1. 2. 3. 4- tetrahydroquinoline- 6- n -carbamic acid 9- fluorenylmethyl ester

To a mixture of the compound described in Example 22b (10.1 g) in anhydrous CH 2 Cl 2 (500 mL) was added dropwise boron tribromide (5.05 mL) and the resulting mixture was stirred for 2.5 h at room temperature. The reaction was quenched with ice-water at 0°C and CH 2 Cl 2 was added. The organic layer was separated, dried and stored at 4°C for 20 hours. The solids formed were collected by filtration and dried in vacuo to give the crude product which was used without further purification. Yield: 12.5 g.

(d) . 1- Acetyl- 6- amino- 2, 2, 4- trimethyl- 4- r4-(3- morpholin- 4- yl- propoxy)- phenyl- 1, 2, 3, 4-tetrahydroquinoline

A mixture of the compound described in Example 22c (1.0 g), Cs 2 CO 3 (1.8 g), 4-(3-chloropropyl)morpholine (330 mg) and DMF (5 mL) was stirred at 60 °C for 18 h .

Water was added and the mixture was extracted with CH 2 Cl 2 . The organic layer was dried and concentrated in vacuo. The title compound was purified by chromatography on silica gel using CH 2 Cl 2 / 2% concentrated ammonia in MeOH = 1/0 => 9/1 (v/v) as the eluent.

Yield 527 mg

(e). / V-{ 1- acetvl- 2, 2, 4- trimethvl- 4- r4-( 3- morpholine- 4- vl- propoxyv)- phenvl1- 1, 2, 3, 4- tetrahydro- quinolin- 6- vl) - 5-bromo-2-methylamino-benzamide

General Procedure C: To a solution of the compound described in Example 22d (132 mg), 5-bromo-2-methylamino benzoic acid (101 mg) and DIPEA (255 µl) in CH2Cl2 (3 mL) was added HATU (166 mg) at room temperature. The reaction mixture was stirred for 18 hours at room temperature. Ethyl acetate (15 mL) and 2 M aqueous NaOH (15 mL) were added. The organic layer was separated and washed with 2 M aqueous NaOH (10 mL) and water (15 mL), dried and concentrated in vacuo. The title compound was purified by preparative HPLC (Method A).

Yield: 69.8 mg; MS-ESI: [M+H]<+>= 663.4; HPLC: Rt = 14.65 min (method 3).

Example 23

A/-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 3- morpholine- 4- vl- propoxyv)- phenvn- 1, 2, 3, 4- tetrahydroquinoline- 6- vl)- 3 , 5- dichloro- 2, 6- dimethoxybenzamide

According to General Procedure C, the compound described in Example 22d (132 mg) was acylated with 3,5-dichloro-2,6-dimethoxybenzoic acid (110 mg), DIPEA (255 µl) and HATU (166 mg) in CH 2 Cl 2 (3 ml). The title compound was purified by preparative HPLC (Method A).

Yield: 68.3 mg; MS-ESI: [M+H]<+>= 684.3; HPLC: Rt = 13.45 min (method 3).

Example 24

Biphenyl- 4- carboxylic acid H- acetyl- 4-( 4-{ 2-[( furan- 2- ylmethyl)- aminol- ethoxy) phenyl)-2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro - quinoline-6-vn-amide

(a) . ( 4-{ 1- acetvl- 6- r( biphenvl- 4- carbovl) amino1- 2, 2, 4- trimethylvl- 1, 2, 3, 4- tetrahydro-quinoline- 4- vl)- phenoxv) acetic acid- ethyl ester

A mixture of the compound described in Example 1f (1 g), ethyl bromoacetate (220 µl), K 2 CO 2 (850 mg) and acetone (25 ml) was stirred for 6 hours at 50°C. The solids were removed by filtration and the filtrate was concentrated in vacuo to give the product which was used without further purification in the next step.

Yield: 1.2 g

(b) . Biphenyl- 4- carboxylic acid { 1- acetyl- 4-[ 4-( 2- hydroxy ) phenyl- 2, 2, 4- trimethyl-1, 2, 3, 4- tetrahydroquinolin- 6- yl}-amide

To a solution of the compound described in Example 24a (1.2 g) in THF (10 ml) at 0°C was carefully added LiALH 4 (78 mg) and the resulting mixture was stirred for 3 hours at room temperature. Ethyl acetate (50 mL) was added dropwise, followed by water (50 mL). The aqueous layer was separated and extracted with ethyl acetate (50 mL) and the combined organic fractions were washed with brine. The organic layer was dried and concentrated in vacuo to give the product which was used without further purification in the next step.

Yield: 1 g

(c) . Methanesulfonic acid 2-( 4-{ 1- acetvl- 6- r( biphenvl- 4- carbovl) amino1- 2, 2, 4- trimethvl-1, 2, 3, 4- tetrahydroquinoline- 4- vl} phenoxyl) ethyl ester

To a solution of the compound described in Example 24b (1 g) and DIPEA (1.7 mL) in CH 2 Cl 2 (15 mL), a solution of methanesulfonyl chloride (310 µl) in CH 2 Cl 2 (5 mL) was added dropwise. After 2 hours, water was added, the organic layer separated, dried and concentrated in vacuo. The title compound was purified by chromatography on silica gel using heptane/ethyl acetate = 9/1 => 1/1 (v/v) as the eluent.

Yield: 870 mg

(d) . Biphenyl- 4- carboxylic acid ri- acetyl- 4-( 4-{ 2- r( furan- 2- ylmethyl)- amino1- ethoxy)- phenvl)- 2, 2, 4- trimethylvl- 1, 2, 3, 4- tetrahydroquinoline-6-vn-amide

General Procedure D: To a solution of the compound described in Example 24c

(87 mg) in CH 3 CN (5 mL) was added 2-furfurylamine (107 mg) and the resulting mixture was stirred at 70°C for 18 h. The mixture was concentrated in vacuo and the product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 47 mg (TFA salt); MS-ESI: [M+H]<+>= 628.6; HPLC: Rt = 11.53 min (method 4).

Example 25

Biphenyl-4-carboxylic acid (1-acetyl-4-(4- r2-(2-hydroxy-1,1-dimethyl-ethylamino)-ethoxy1-phenyl)-2,2,4-trimethyl-1,2,3, 4-tetrahydro-quinolin-6-yl)-amide

According to general procedure D, the compound described in Example 24c (87 mg) was treated with 2-amino-2-methyl-propan-1-ol (100 mg) in CH 3 CN (5 mL). The title compound was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt. Yield: 21 mg (TFA salt); MS-ESI: [M+H]<+>= 619.8; HPLC: Rt = 10.95 min (method 4).

Example 26

Biphenyl- 4- carboxylic acid triacetyl- 2, 2, 4- trimethyl- 4-( 4-{ 2- r( pyridine- 3- trimethyl) aminol-ethoxy)- phenyl)- 1, 2, 3, 4- tetrahydro - quinoline-6-vn-amide

According to general procedure D, the compound described in Example 24c (87 mg) was treated with 3-aminomethylpyridine (119 mg) in CH 3 CN (5 mL). The title compound was purified by preparative HPLC (method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt.

Yield: 40 mg (TFA salt); MS-ESI: [M+H]<+>= 639.4; HPLC: Rt = 10.15 min (method 4).

Example 27

Biphenyl- 4- carboxylic acid ( 1- acetyl- 4-{ 4- r2-( 2- hydroxy- ethylamino)- ethoxy- 1- phenyl)-2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydroquinoline- 6-vl)-amide

According to general procedure D, the compound described in Example 24c (100 mg) was treated with ethanolamine (100 mg) in CH 3 CN (5 mL). The title compound was purified by preparative HPLC (Method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt

Yield: 50 mg (TFA salt); MS-ESI: [M+H]<+>= 592.6; HPLC: Rt = 10.32 min (method 1).

Example 28

Biphenyl- 4- carboxylic acid ( 1- acetyl- 4-{ 4-[ 2-( 2- amino- ethylamino)- ethoxy- phenyl}- 2, 2, 4-trimethyl- 1, 2, 3, 4- tetrahydroquinoline - 6-vl)-amide

According to general procedure D, the compound described in Example 24c (100 mg) was treated with ethylenediamine (110 mg) in CH 3 CN (5 mL). The title compound was purified by preparative HPLC (Method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt

Yield: 45 mg (TFA salt); MS-ESI: [M+H]<+>= 591.4; HPLC: Rt = 7.04 min (method 1).

Example 29

Biphenyl- 4- carboxylic acid { 1 - acetyl- 2, 2, 4- trimethyl- 4- r4-( 2- piperazine- 1 - vl- ethoxy) phenyl-1, 2, 3, 4- tetrahydro- quinolin- 6- vl )- amide

According to general procedure D, the compound described in Example 24c (100 mg) was treated with piperazine (140 mg) in CH 3 CN (5 mL). The title compound was purified by preparative HPLC (Method A) and lyophilized from a mixture of CH 3 CN and water containing TFA, to give the corresponding TFA salt

Yield: 95 mg (TFA salt); MS-ESI: [M+H]<+>= 617.6; HPLC: Rt = 9.54 min (method 1).

Example 30

Morpholine- 4- carboxylic acid ( 3-{ 4-[ 1- acetyl- 6-( 3, 5- dichloro- 2, 6- dimethoxy-benzoylamino)- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydroquinoline-4-n-phenoxy)-propyl)-amide (a) . Morpholine- 4- carboxylic acid ( 3-{ 4-[ 1 - acetyl- 6- amino- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro- quinolin- 4- n-phenoxy)- propyl)- amide

Following the same procedure described in Example 22d, the compound described in Example 22c (1.0 g) was alkylated (with accompanying removal of the Fmoc protecting group) with morpholine-4-carboxylic acid (3-chloropropyl)amide (448 mg) using Cs 2 CO 3 (1.8 g) in DMF (5 mL). The title compound was purified by chromatography on silica gel using CH 2 Cl 2 / 2% concentrated ammonia in MeOH = 1/0 => 9/1 (v/v) as the eluent.

Yield: 894 mg.

(b) . Morpholine- 4- carboxylic acid ( 3-{ 4- triacetyl- 6-( 3, 5- dichloro- 2, 6- dimethoxy-benzoylamino)- 2, 2, 4- trimethyl- 1, 2, 3, 4- tetrahydro - quinoline-4-vn-phenoxy)-propyl)-amide According to general procedure C, the compound described in Example 30a (228 mg) was acylated with 3,5-dichloro-2,6-dimethoxybenzoic acid (230 mg), DIPEA (558 ul) and

HATU (609 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A).

Yield: 102 mg; MS-ESI: [M+H]<+>= 727.4; HPLC: Rt = 22.37 min (method 2).

Example 31

/ V-{ 1- acetvl- 2. 2. 4- trimethvl- 4- r4-( 2- morpholin- 4- v- ethoxy)- phenvn- 1. 2. 3. 4- tetrahydroquinolin- 6- yl)- 3 , 5-dibromobenzamide

(a) . 1- acetvl- 6- amino- 2, 2, 4- trimethyl- 4-[ 4-( 2- morpholine- 4- v- ethoxy)- phenvn- 1, 2, 3, 4-tetrahydroquinoline

A mixture of the compound described in Example 22c (1.0 g), Cs 2 CO 3 (1.8 g), N-(2-chloroethyl)-morpholine hydrochloride (375 mg) and DMF (5 mL) was stirred at 60°C in 18 hours. The reaction did not reach completion and additional amounts of Cs 2 CO 3 and N-(2-chloroethyl)-morpholine hydrochloride were added. After the reaction was complete, water was added and the mixture was extracted with CH 2 Cl 2 . The organic layer was dried and concentrated in vacuo. The title compound was purified by chromatography on silica gel using CH 2 Cl 2 / 2% concentrated ammonia in MeOH = 1/0 => 9/1 (v/v) as the eluent.

Yield: 905 mg.

(b) . A/-{ 1- acetvl- 2, 2, 4- trimethvl- 4- r4-( 2- morpholine- 4- vl-ethoxv)- phenvn- 1, 2, 3, 4- tetrahdro- quinolin- 6- vl) - 3, 5-dibromobenzamide

According to General Procedure C, the compound described in Example 31a (157 mg) was acylated with 3,5-dibromobenzoic acid (150 mg), DIPEA (313 µl) and HATU (204 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 71 mg (TFA salt); MS-ESI: [M+H]<+>= 700.2; HPLC: Rt = 16.12 min (method 2).

Example 32

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholin- 4- v- ethoxy)- phenvn- 1, 2, 3, 4- tetrahydroquinolin- 6- yl)- 2 - chlorbenzamide

According to General Procedure C, the compound described in Example 31a (150 mg) was acylated with 2-chlorobenzoic acid (81 mg), DIPEA (299 µl) and HATU (195 mg) in CH 2 Cl 2 (6 mL). The title compound was purified by preparative HPLC (Method A). Yield: 162 mg (TFA salt); MS-ESI: [M+H]<+>= 576.4; HPLC: Rt = 9.37 min (method 2).

Example 33

/ V-( 1- acetvl- 2. 2. 4- trimethvl- 4- r4-( 2- morpholine- 4- vl- ethoxv)- phenvn- 1. 2. 3. 4- tetrahydroquinoline- 6- vl)- 3 , 5-dimethylbenzamide

According to General Procedure C, the compound described in Example 31a (200 mg) was acylated with 3,5-dimethylbenzoic acid (103 mg), DIPEA (399 µl) and HATU (260 mg) in CH 2 Cl 2 (7.5 mL). The title compound was purified by preparative HPLC (method

A).

Yield: 57.5 mg (TFA salt); MS-ESI: [M+H]<+>= 570.4; HPLC: Rt = 12.62 min (method 2).

Example 34

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholin- 4- v- ethoxy)- phenvn- 1, 2, 3, 4- tetrahydroquinolin- 6- yl)- 2 , 5-dichlorobenzamide

According to General Procedure C, the compound described in Example 31a (200 mg) was acylated with 2,5-dichlorobenzoic acid (131 mg), DIPEA (399 µl) and HATU (260 mg) in CH 2 Cl 2 (7.5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 130 mg (TFA salt); MS-ESI: [M+H]<+>= 610.2; HPLC: Rt = 11.70 min (method 2).

Example 35

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholin- 4- v- ethoxy)- phenvn- 1, 2, 3, 4- tetrahydroquinolin- 6- yl)- 5 - methyl-2-nitrobenzamide

According to General Procedure C, the compound described in Example 31a (157 mg) was acylated with 5-methyl-2-nitrobenzoic acid (97.3 mg), DIPEA (313 µl) and HATU (204 mg) in CH 2 Cl 2 (5 mL) . The title compound was purified by preparative HPLC (Method A).

Yield: 80 mg (TFA salt); MS-ESI: [M+H]<+>= 601.4; HPLC: Rt = 9.95 min (method 2).

Example 36

/ V-{ 1- aceWI- 2, 2. 4- trimethvl- 4- r4-( 2- morpholine- 4- vl- ethoxv)- phenvl1- 1, 2, 3, 4- tetrahquinolin- 6- vl)- benzamide

According to General Procedure C, the compound described in Example 31a (157 mg) was acylated with benzoic acid (65.6 mg), DIPEA (313 µl) and HATU (204 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A).

Yield: 59 mg (TFA salt); MS-ESI: [M+H]<+>= 542.4; HPLC: Rt = 9.99 min (method 2).

Example 37

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholine- 4- vl- ethoxv)- phenvn- 1, 2, 3, 4- tetrahydroquinoline- 6- vl)- 4 - tert-butyl-benzamide

According to General Procedure C, the compound described in Example 31a (161 mg) was acylated with 4-tert-butylbenzoic acid (99 mg), DIPEA (322 µl) and HATU (210 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 80 mg (TFA salt); MS-ESI: [M+H]<+>= 598.2; HPLC: Rt = 15.39 min (method 2).

Example 38

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholine- 4- vl- ethoxv)- phenvn- 1, 2, 3, 4- tetrahydroquinoline- 6- vl)- 2 , 3-dichlorobenzamide

According to General Procedure C, the compound described in Example 31a (161 mg) was acylated with 2,3-dichlorobenzoic acid (106 mg), DIPEA (322 µl) and HATU (210 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 113 mg (TFA salt); MS-ESI: [M+H]<+>= 610.2; HPLC: Rt = 11.42 min (method 2).

Example 39

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholin- 4- v- ethoxy)- phenvn- 1, 2, 3, 4- tetrahydroquinolin- 6- yl)- 4 - bromobenzamide

According to General Procedure C, the compound described in Example 31a (260 mg) was acylated with 4-bromobenzoic acid (179 mg), DIPEA (517 µl) and HATU (338 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A). Yield: 127 mg (TFA salt); MS-ESI: [M+H]<+>= 620.2; HPLC: Rt = 12.24 min (method 2).

Example 40

/ V-( 1- acetvl- 2. 2. 4- trimethvl- 4- r4-( 2- morpholine- 4- vl- ethoxv)- phenvn- 1. 2. 3. 4- tetrahydroquinoline- 6- vl)- 4 - methoxy-3-methylbenzamide

According to General Procedure C, the compound described in Example 31a (260 mg) was acylated with 4-methoxy-3-methylbenzoic acid (148 mg), DIPEA (517 µl) and HATU (338 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A).

Yield: 158 mg (TFA salt); MS-ESI: [M+H]<+>= 586.2; HPLC: Rt = 11.49 min (method 2).

Example 41

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholin- 4- v- ethoxy)- phenvn- 1, 2, 3, 4- tetrahydroquinolin- 6- yl)- 4 - dimethylaminobenzamide

According to General Procedure C, the compound described in Example 31a (260 mg) was acylated with 4-dimethylaminobenzoic acid (147 mg), DIPEA (517 µl) and HATU (338 mg) in CH 2 Cl 2 (5 mL). The title compound was purified by preparative HPLC (Method A).

Yield: 95 mg (TFA salt); MS-ESI: [M+H]<+>= 585.2; HPLC: Rt = 9.53 min (method 2).

Example 42

/ V-{ 1- acetvl- 2, 2. 4- trimethvl- 4- r4-( 2- morpholine- 4- vl- ethoxv)- phenvn- 1, 2, 3, 4- tetrahydroquinoline- 6- vl)- 3 - trifluoromethylbenzamide

To a solution of the compound described in Example 31a (260 mg) and pyridine (500 µl) in toluene (4.5 ml) was added 3-(trifluoromethyl)benzoyl chloride (185 mg). Ethyl acetate (15 mL) and water (15 mL) were added. The organic layer was separated and washed with water (15 mL), dried and concentrated in vacuo. The title compound was purified by preparative HPLC (Method A).

Yield: 200 mg (TFA salt); MS-ESI: [M+H]<+>= 610.2; HPLC: Rt = 13.23 min (method 2).

Example 43

/ V-( 1- acetvl- 2. 2. 4- trimethvl- 4- r4-( 2- morpholine- 4- vl- ethoxv)- phenvn- 1. 2. 3. 4- tetrahydroquinoline- 6- vl)- 3 - nitrobenzamide

To a solution of the compound described in Example 31a (260 mg) and pyridine (500 µl) in toluene (4.5 ml) was added 3-nitrobenzoyl chloride (165 mg). Ethyl acetate (15 mL) and water (15 mL) were added. The organic layer was separated and washed with water (15 mL), dried and concentrated in vacuo. The title compound was purified by preparative HPLC (Method A).

Yield: 167 mg (TFA salt); MS-ESI: [M+H]<+>= 587.4; HPLC: Rt = 10.28 min (method 2).

Example 44

CHO-FSH in vitro bioactivity

FSH activity of compounds was tested in Chinese hamster ovary (CHO) cells stably transfected with the human FSH receptor and co-transfected with a cAMP responsive element (CRE)/promoter that directs expression of a firefly luciferase reporter gene. Binding of ligand to the Gs-coupled FSH receptor will result in an increase in cAMP, which in turn will induce an increased transactivation of the luciferase reporter construct. To test antagonistic properties, recombinant FSH at a concentration that induces approximately 80% of the maximal stimulation of cAMP accumulation in the absence of test compound was added (rec-hFSH; 10 mU/ml). The luciferase signal was quantified using a luminescence counter. For test compounds, EC5 excess (concentration of test compound causing half-maximal (50%) stimulation or reduction) was calculated. For that purpose, the software GraphPad PRISM, version 3.0 (GraphPad software Inc., San Diego) was used.

Compounds of all examples showed an EC50 (IC50) value of less than 10"<5>M in either an agonistic or an antagonistic assay set-up or both. The compounds of Examples 3, 4, 7, 10-13, 16, 36, 37, 39, 41 and 42 showed an EC50(IC50) of less than 10"<7>M in at least one of the experiments.

Claims (6)

1. Tetrahydroquinoline derivative according to Formula 1, or a pharmaceutically acceptable salt thereof, wherein R<1> and R<2> are Me; R<3> is morpholinylcarbonylamino or R<5->(2-4C)alkyl; R<4> is biphenyl or phenyl optionally substituted with halogen, (1-6C) alkoxy, (1-6C) alkyl, NO 2 , (di)(1-6C) alkylamino or CF 3 ; R<5> is (di)(1-4C)alkylamino, morpholinyl, piperazinyl, pyridyl, piperidinyl optionally substituted with (1-6C)alkyl, azepanyl, aminocarbonyl, (2-7C)alkenylaminocarbonyl, (1-6C)alkylamino, pyridyl-(1-4C)alkylaminocarbonyl, furyl-(1-4C)alkylaminocarbonyl, (1-6C)alkoxy-(1-6C)alkylaminocarbonyl, phenyl-(1-4C)alkylaminocarbonyl, (1-6C)alkylaminocarbonyl, morpholinylcarbonyl, furyl-(1-4C)alkylamino, hydroxy-(1-6C)alkylamino, pyridyl-(1-6C)alkylamino, amino-(1-6C)alkylamino or morpholinylcarbonylamino. 2. Derivative according to claim 1, where R<5> is (1-4C)alkylamino. 3. Derivative according to claims 1-2, where R<4> is phenyl. 4. Pharmaceutical preparation comprising the tetrahydroquinoline derivative according to any of claims 1-3 and pharmaceutically suitable additives. 5. Tetrahydroquinoline derivative according to claims 1-3 for use in therapy. 6. Use of a tetrahydroquinoline derivative according to any one of claims 1-3 or a pharmaceutically acceptable salt or solvate thereof for the preparation of a medication for fertility regulation.