ZA200504776B - Tetrahydroquinoline derivatives and their use as FSH receptor modulators - Google Patents

Description

. Tetrahydroquinoline derivatives .

The invention relates to @ compound having FSH receptor modulatory activity, in particular a tetrahydroquin «line derivative, to a pharmaceutical composition containing the same, as well as the use of said compound in medical therapy.

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. The hypophyseal gonadotropin FSH (follicle stimulating hormone) for example plays a pivotal role in the stimulation of follicle development and maturation whereas LH (luteinizing haormone) induces ovulation (Sharp, R.M. Clin Endocrinol. 33:787-807, 1990; Dormrimmgton and Armstrong, Recent Prog. Horm. Res. 35:301- 342,1979). Currently, FSHL is applied clinically, in combination with LH or hCG, for ovarian stimulation i.e. owarian hyperstimulation for in vitro fertilisation (IVF) and induction of ovulation in imafertile anovulatory women (Insler, V., Int. J. Fertility 33:85- 97, 1988, Navot and Roserawaks, J. Vitro Fert. Embryo Transfer 5:3-13, 1988), as well as for male hypogonadism aud male infertility.

The gonadotropin FSH is released from the anterior pituitary under the influence of gonadotropin-releasing hosrmone and oestrogens, and from the placenta during pregnancy. In the female, FSH acts on the ovaries promoting development of follicles and is the major hormone regulating secretion of oestrogens. 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 females and for some types of failure of spermatogenesis in males. Gonadotropins destined 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). o Alternatively, they can be prepared as recombinant gonadotropins. Recombinant human FSH is available «<commercially and is being used in assisted reproduction * (Olijve et al. Mol. Hum. Resprod. 2:371, 1996; Devroey et al. Lancet 339:1170, 1992).

The actions of the FSH hormone are mediated by a specific plasma membrane receptor that is a member 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 highly specific target in the ovarian follicle growth process and is exclusively expressed in the ovary. Blocking this receptor or inhibiting the signaling which is normally induced after FSHI-mediated receptor activation will disturb follicle development and thus ovulation and fertility. Low molecular weight FSH antagonists could therefore form the basis for mew contraceptives. Such FSH antagonists could give rise to diminished follicle development (no ovulation) with still sufficient estrogen production left 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 present invention describes the preparation of low molecular weight hormone analogs that selectively have modulatory activity on the FSH receptor. The compounds of the invention can either be used as (partial) agonists or (partial) antagonists of the

FSH-receptor.

Thus, it has now been found, thaat the following class of tetrahydroquinoline compounds of formula I or pharmaceutically acceptable salts thereof, have FSH- modulatory activity:

\ o-R3

H ol

R2 © N° R1 0?

Formula I wherein

R' and R? are H, MCe;

Ris (2-6C)heter-ocycloalkyl(1-4C)alkyl, (2-5C)heteroaryl(1-4C)alkyl, (6C)aryl(1— 4C)alkyl, (1-4C)(di)alkylaminocarbonylamino(2-4C)alkyl, 2— 6C)heterocycloalky-lcarbonylamino(2-4C)alkyl, R>-(2-4C)alkyl or R’-carbonyl(1— 4C)alkyl;

R* is (2-5C)heteroaryl, (6C)aryl, (3-8C)cycloalkyl, (2-6C)heterocycloalkyl or (a- 6C)alkyl

R® is (di)(1-4C)allk=ylamino, (1-4C)alkoxy, amino, hydroxy, (6C)arylamino, (d)(3— 4C)alkenylamino, (2-5C)heteroaryl(1-4C)alkylamino, (6C)aryl(1-4C)alkylamino, (di)[(1-4C)alkoxy(2—4C)alkyl]amino, (di)[(1-4C)alkylamino(2-4C)alkyljamino, (di)[amino(2-4C)alk=ylJamino or (di)[hydroxy(2-4C)alkyl]amino.

The compounds acceording to the present invention modulate the FSH receptor function and can be used foer the same clinical purposes as native FSH if they behave like agonists, with the a«dvantage that they display altered stability properties and may be administered differently. If they block the FSH receptor they can be used e.g. as a , 20 contraceptive agent.

Thus, the FSH-receptor modulators of the present invention may be used for treating 8 infertility, for contra-ception and for treatment of hormone-dependent disorders such as breast cancer, prostate cancer, and endometriosis.

. a. . The following teerms are intended to have the indicated meanings demnoted below as used in the specification and claims.

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

The term (2-4CDalkyl as used herein means a branched or unbranched alkyl group having 2-4 carbo-n atoms, being ethyl, propyl, isopropyl, butyl, sec-butyl and tert-butyl.

The term (1-6CDalkyl as used herein means a branched or unbranch ed 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 being the most preferred.

The term (di)(1-<3C)alkylamino as used herein means an amino group, nnonosubstituted or disubstituted with alkyl groups, each of which contains 1-4 carbon atoms and has the same meaning as previously defined.

The term (di) 1-4C)alkenylamino as used herein means an amino group, monosubstituted or disubstituted with alkenyl groups, each of whick contains 2-4 carbon atoms su-ch as allyl and 2-butenyl, and has the same meanings as previously defined.

The term (3-8C)cycloalkyl as used herein means a cycloalkyl group having 3-8 carbon atoms, being cxyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cy=cloheptyl and cyclooctyl. (3-6C)cycloalkyl groups are preferred.

The term (2-6C)meterocycloalkyl as used herein means a heterocycloalky=1 group having 2-6 carbon atomss, preferably 3-5 carbon atoms, and at least including ©ne heteroatom selected from N, O and/or S, which may be attached via a heteroatom if feasible, or a carbon atom. Preferred heteroatoms are N or O. The hetrocycloalkyl group may be ’ substituted with z3 methyl or ethyl group at a carbon atom, or a heteroatom if feasible.

Most preferred Theterocycloalkyl groups are piperidinyl, piperazinyl., morpholinyl, : pyrrolidinyl and RM -methyl-2-piperidinyl.

. The term (1-4C)alkoxy as used herein means an alkoxy group having 1-4 carbon atoms, the alkyl moiety having the same meaning as previously defined . (1-2C)Alkoxy groups are preferred.

The term (6C)aryl as used herein means a phenyl group, which maw optionally be substituted with one or more substituents selected from hydroxy, amineo, iodo, bromo, chloro, fluoro, nitro, trifluoromethyl, cyano, phenyl, (1-4C)alkyl, (1-4C)alkoxy or (1- -<4C)(di)alkylamino, the alkyl, alkoxy and (di)alkylamino moieties hamving the same ameaning as previously defined, for example phenyl, 3,5-dibromophemyl, 4-biphenyl, 3,5-dichlorophenyl, 3-bromo-6-methylamino-phenyl, 3-chloro-2,6-di=methoxyphenyl and 3,5-dimethylphenyl. “The term (2-5C)heteroaryl as used herein means a substituted or unsubstituted aromatic £roup having 2-5 carbon atoms, at least including one heteroatom seleected from N, O and/or S, like imidazolyl, pyridyl, pyrimidy), thienyl or furyl. The subsstituents on the

Theteroaryl group may be selected from the group of substituents listed £or the (6C)aryl group. The heteroaryl group may be attached via a carbon atom or a heteroatom, if feasible. Preferred heteroaryl groups are thienyl, furyl and pyridyl. "The term (2-6C)heterocycloalkyl(1-4C)alkyl as used herein means a heeterocycloalkyl group having 2-6 carbon atoms, connected to an alkyl group having 1-4- carbon atoms, tthe heterocycloalkyl group and the alkyl group having the same meanin_g as previously «defined. "The term (2-6C)heterocycloalkylcarbonylamino as used heredn means a

Theterocycloalkyl group having 2-6 carbon atoms, connected to the carbomyl moiety of a carbonylamino group, the heterocycloalkyl group having the samee meaning as previously defined.

The term (2-6C)heterocycloalkylcarbonylamino(2-4C)alkyl as used h-erein means a , Eaeterocycloalkylcarbonylamino group of which the heterocycloalkyl moiety contains 2- © carbon atoms, connected via the amino group to an alkyl group having 2-4 carbon ) atoms, the beterocycloalkylcarbonylamino group and the alkyl group having the same rneaning as previously defined.

Whe term (di)(1-4C)alkylaminocarbonyl as used herein means a (di)alky/lamino group, the alkyl group(s) of which having 1-4 carbon atoms, connected via the &mino group to

. a carbonyl group, the (di)alkylamino group having the samme meaning as previously defined.

The term (3-8C)cycloalkylaminocarbonyl as used herein mmeans a cycloalkyl group maving 3-8 carbon atoms, connected to the amino moiety of an aminocarbonyl group, 5s the cycloalkyl group having the same meaning as previously defined.

The term (di)(1-4C)alkylaminocarbonylamino as used here-in means a (di)alkylamino group, the alkyl group(s) of which having 1-4 carbon atomss, connected via the amino group to the carbonyl moiety of a carbonylamino growp, thus providing a urea fanctionality, the (di)alkylamino group having 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) of which having 1-4 carbon atoms, connected via the amino group to an alkyl group hawing 2-4 carbon atoms, the (di)alkylaminocarbonylamino group and the alkyl group ha-ving the same meaning as previously defined.

The term (2-5C)heteroaryl(1-4C)alkyl as used herein 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 gpbenyl group, optionally substituted with one or more substituents selected from the group of substituents listed for the (6C)aryl group, connected to an alkyl group having 1-4 carbon atoms, the aryl gxoup and the alkyl group having the same meaning as previously defined.

T he term (6C)arylamino 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 an amino group, the aryl groupe 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 moiety of an allkylamino group having 1- 4 carbon atoms, the aryl group and the alkylamino group ha.ving the same meaning as pweviously 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 more substituents selected from the group of substituents listed for the (6C)aryl group, connected to the alkyl moiety of an alkylamino group having 1-4 carbon atoms, the heteroaryl group and the alkylamino group having 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 alkyl group having the same meaning as previously desfined

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

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

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

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

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

The term hydroxy(2-4C)alkyl as used herein means an hydroxyalkyl group having 2-4 ' carbon atoms, the alkyl moiety having the same meaning ass previously defined.

The term (di){bydroxy(2-4C)alkyllamino as used herein means an amino group, monosubstituted or disubstituted with hydroxyalkyl groups, having 2-4 carbon atoms and having the same meaning as previously defined.

The term R’-(2-4C)alkyl as used herein means a R® group attached to an alkyl moiety having 2-4 carbon atoms which has the same meaning as previously defined.

The term R*-carbonyl-(1-4C)alkyl as used therein means a R® group attached to the carbonyl moiety of a carbonylalkyl group, the alkyl moiety having 1-4 carbon atoms and having 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 for the tissues of humans and lower animals without undue toxicity, irritation, allergic response and the like, and are commensurate with a reasonable benefit/risk ratio. Pharmaceutically acceptable salts are well known in the art. They may be obtained during the final isolation and purification of the compounds of 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, rnalonic acid, fumaric acid, glycolic acid, succinic acid, propionic acid, acetic acid, methanesulfonic acid, and the like. If present, an acid function can be reacted with an organic or a mineral base, like sodium hydroxide, potassium hydroxide or lithium hy~droxide.

The invention thus relates to the compounds of Formula I as defined here above.

In another embodiment the invention provides compounds according to Formula I wherein R! and R? are Me.

The invention also relates to compounds of formula I, wherein R® is (2- } 6C)heterocycloalkyl(1-4C)alkyl, (2-5C)heteroaryl(1-4C)alkyl, 2- 6C)heterocycloalkylcarbonylamino(2-4C)alkyl, R’-(2-4C)alkyl, R>-carbonyl(1- ) 4C)alkyl.

In another aspect the invention concerns compmunds according to Formula I wherein

Ris (2-6C)heterocycloalkyl(1-4C)alkyl, (2-5C)hheteroaryl(1-4C)alkyl, R’-(2-4C)alkyl,

R’-carbonyl(1-4C)alkyl.

In yet another aspect the invention relates to compounds according to Formula I wherein R? is (2-6C)heterocycloalkyl(1-4C)alky=l, (2-5C)heteroaryl(1-4C)alkyl or R’- (2-4C)alkyl.

In another aspect the invention relates to compounds according to Formula I wherein

R? is (2-6C)heterocycloalkyl(1-4C)alkyl.

According to yet another embodiment of the in-vention the heterocycloalkyl group in heterocycloalkyl(1-4C)alkyl in R® according to Frormula I consists of 4, 5 or 6 C atoms and the heteroaryl group in heteroaryl(1-4C)alkyll in R? consists of 3, 4 or 5 C atoms.

In another embodiment the invention relates to» compounds according to Formula I, wherein R* is (6C)aryl.

In yet another embodiment the invention providess compounds of Formula I wherein rR’ is (di)(1-4C)alkylamino, amino, (di)(3-4C)alkenylamino, (2-5C)heteroaryl(1- : 4Calkylamino, (6C)aryl(1-4C)alkylamino, (di)[(1-4C)alkoxy(2-4C)alkyl]amino, (di)[(1-4C)alkylamino(2-4C)alkyl]amino, (di)[amino(2-4C)alkylJamino, (di)[hydroxy(2-4C)alkylJamino.

In another aspect the invention relates to compounds according to Formula I wherein

R} is (di)(1-4C)alkylamino, (2-5C)heteroaryl(1 -4C)alkylamino, (di)[(1-4C)alkoxy(2- 4C)alkylJamino, (di)[(1-4C)alkylamino(2—4C)alkylJamino, (di){amino(2- 4C)alkylJamino or (di)[hydroxy(2-4C)alkyllamimo.

In another aspect the invention relates to compounds according to Formula I wherein

R® is (di)(1-4C)alkylamino, amino, (di)(3-4-C)alkenylamino, (2-5C)heteroaryl(1- 4C)alkylamino, (6Caryl(1-4C)alkylamino.

Another aspect of the invention are compounds according to Formula I wherein R’is i (di)(1-4C)alkylamino or amino.

In yet another aspect of the invention, there awe provided compounds according to

Formula I wherein R? is (di)(1-4C)alkylamino.

Yet another aspect of” the invention concerns compounds wherein all specific definitions of the group s R! through R® as defined here above are combined in the compound of Formula I.

Suitable methods for the preparation of the compounds of the invention are outlined below. 0 RB ® B oc Bec foe

H Hi HN XN HN = “Nr TV ” . R1

N

0 @® Ri NH, Ne R2 i o © Io ma Rk IV-aRI,R? = Me = = 1 R2= [-aR!R?=Me IV-bRLR?=H

Ib RLR2=H

The compounds of the present invention with formula I-a can be prepared starting with the well-documented Skraup reaction. Performing this reaction on N-fert- butoxycarbonyl (N-Boe) protected 1,4-phenylenediamine (II) gives 1,2- dihydroquinoline derivative Ill-a.

Related Skraup cyclocon densation reactions are found in literature: A. Knoevenagel,

Chem. Ber, 54:1726, 192.1; RL. 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.Bs. Marschke, D.E. Mais, M.M. Gottardis and T.K. Jones, J.

Med. Chem. 41:303, 1998. } The abovementioned reaction is typically conducted at elevated temperature in acetone or mesityl oxide in the presence of iodine or protic acid such as hydrochloric acid, p- toluenesulfonic acid or =iqueous hydrogen iodide. Alternatively, the compound of formula III-a can be prepared by reacting compound II with acetone in the presence of

MgSO0,, 4-tert-butylcatechol and iodine (L.G. Hamann, R.I. Higuchi, L. Zhi, J.P.

Edwards and X.-N. Wang, J. Med. Chem, 41:623, 1998). In yet another procedure, the reaction can be performed in acetorze using lanthanide triflates (e.g. scandium triflate) as catalysts. In this case, the reacti«on 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 N-Boc-1,4-phenylenediamine II by reaction with methyl vinyl ketone. Related cyclizations are described in Untited

States Patent 2,686,182 (Badische Anilin- & Soda-Fabrik Aktiengesellschaft).

Subsequent 1-N-acetylation of the «compounds of formula II-a-b can be carried out using standard conditions. In a typical experiment, compounds of formula IlI-a-b are heated under reflux in acetic anhydri de or reacted in a solvent such as dichloromethane, tetrahydrofuran, toluene or pyridine with acetyl chloride in the presence of a base such as N,N-diisopropylethylamine, trieth_ylamine or sodium hydride to give the 1-N-acetyl- 4-methyl-1,2-dihydroquinoline deriv-atives of formula IV-a-b.

IV-a-b OMe OH

Rp “p° @ Rp ()

HN x HN ~ HN HN

Rl R1 — © R1 — C) R1 ke Yoke ke Y ke

IPN A A PN

V-aRL,R2=Me VI-aR!,R? =Me VH-aR!R?2=Me VII-aR!R?=Me

V-bRIR2=H VIbRLR2 =H VIIbRIR2=H VII-b RLRZ=H

Standard cleavage of the Boc protecstive group under conditions well known to those skilled in the art affords the 6-amino -1,2-dihydroquinoline derivatives of formula V-a- ° b. This reaction is typically conclucted in dichloromethane in the presence of trifluoroacetic acid.

Subsequent 6-N-acylation of the commpounds of formula V-a-b can be carried out using standard conditions to give compoumds of general formula VI-a-b, wherein R* 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*-c(0)-Cn or acid anhydsride (R*-C(0)-O-C(O)-R") in the presence of a base such as NN- diisopropylethy~lamine, triethylamine, pyridine or sodium hydride to give 6-N—acylated 4-methyl-1,2-dihydroquinoline derivatives of formula VI-a-b. Alternatively, acylation of compounds of general formula V-a-b to give compounds of general formula VI-a-b can also be accomplished by reaction with an appropriate carboxylic acid (R*~C0,H) in the presence of a coupling reagent such as O-(benzotriazol-1-yl)-N,2V,N’N’- tetramethyluromium tetrafluoroborate (TBTU), O-(7-azabenzotriazol-1-yl)-N,2V,N’N’- tetramethyluromium hexafluorophosphate (HATU) or bromotripyrrolidinophosgohonium hexafluorophosgphate (PyBrOP) and a tertiary base, e.g. N,N-diisopropylethyla—mine, in a solvent such zas NV N-dimethylformamide or dichloromethane at ambient or elevated temperature.

Introduction o=f the requisite substituted phenyl group at position 4 of the dihydroquinolince scaffold can be accomplished via Friedel-Crafts alkylation off anisole with the compowmnds of general structure VI-a-b to yield compounds of general formula

VII-a-b. This re=action is typically conducted at elevated temperatures either im anisole or in an appropriate inert solvent such as heptane or hexane with anisole as reagent, under catalysis of a Lewis acid (e.g. AlCl;, AlBr; FeCl; or SnCL). Friedel-Crafis alkylations witha 2,2,4-trimethyl-1,2-dihydroquinolines are described in literature by

B.A. Lugovik, I..G. Yudin and AN. Kost, Dokl. Akad. Nauk SSSR, 170:34Q), 1966;

B.A. Lugovik, L.G. Yudin, S.M. Vinogradova and AN. Kost, Khim. Ge-terosikl.

Soedin, 7:795, 1. 971.

Alternatively, = N-Boc-1,4-phenylenediamine II can be reacted withh 2-(4- methoxyphenym-propene and formaldehyde in acetonitrile at ambient or eelevated temperature, followed by I1-N-acetylation as described previously, to g-ive the compound VII-'b in which R* = O-tert-Bu. Related cyclizations are descr-ibed in literature: J.M. Mellor and G.D. Merriman, Tetrahedron, 51:6115, 1995. Cleavage of the Boc protectiwe group and subsequent acylation of the 6-amino function with. an acyl . halide (R*-C(O)~Cl) as described before gives access to compounds of general structure

VII-b in which RR*is as described previously.

Cleavage Of the aromatic methyl ether in compounds of general formula ViI-a-b affords 4-(4-hydroxyphenyl) substituted tetrahydroquinoline derivatives of general formula VII-a-b, setting the stage for functionalization of the free OF group. oH 0 3

RE3—X * C) Rt — @ Rf

N N

EN N EN ”

IX-a=X=C(l],Br,I

IX-b=X=OH VII-aR!R?=Me la RIR?=Me

VII-bR!R?=H Fb RIR?=H

Demethylat-ion reactions of aromatic methyl ethers are well known to those skilled in the art. In. a typical experiment, demethylation is achieved up-on reaction of a compound «of formula VII-a-b with BBr3 in an inert solvent such as dichloromethane at low to amb-ient temperature to give demethylated compounds of genseral formula VIII- a-b. Alternatively, demethylation can be accomplished upon reactiom of compounds of formula VIN-a-b with BF; Me;S complex at ambient temperature.

Selective (@-alkylation of compounds of general formula VIII-a-b with functionalized alkyl halides of general formula IX-a, leads to the formation of compounds with 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 formmula VIII-a-b in a suitable soRvent such as 14-dioxane, tetrahydrofuran, dichloromethane, acetonitrile, acetone or _N.N-dimethylformamide is treated with a base (e.g. N,N—diisopropylamine, triethylamirie, K,COs, Cs:COs or NaOH) and the appropriate alkylation reagent of general forrmula IX-a, for example benzyl bromide, 3-(dimethylamino)-propyl chloride, 4-(2-chloroeethyl)-morpholine, 2-picolylchloride or 2-chloroacetami de. Alternatively, alkylation c=an be accomplished by the known Mitsunobu-type alkylation. In that case a solution of a compound of general formula VIII-a-b in a suitable so-lvent such as 1,4- dioxane, tetrahydrofuran, or dichloromethane is treated with (resin bound) triphenyl phosphine, diethyl- or di-terz-butyl azodicarboxylate and a functioralized alcohol of general formula IX-b. In principle, both alkylation methods czan be used for all R® groups, but a suitable protective group strategy may be required if R® contains a nucleophilic group such as a secondary amine or a hydroxyl group. Selection of a protective group and deprotection conditions are trivial to those skilled in the art.

Another procedure to obtain compounds of the current invesntion starts with the alkylation of compounds of general formula VIII-a-b with esters eof general formula X.

The alkylation reaction is typically conducted in the presence o f a base such as N,N- diisopropylethylamine or sodium hydride in a suitable soslvent such as NN- dimethylformamide or tetrahydrofuran at ambient or elevated temperature. The ester

A¢ function in the resulting compounds of general formula XI-a-b ia which A = Me or Et can then selectively be reduced under controlled conditions to afford compounds of general formula XIII-a-b using an appropriate reducing agzent 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 gereral formula XIII-a-b

A5 may subsequently be reacted with 4-toluenesulfonyl c=hloride (Ts-Cl) or methanesulfonyl choride (Ms-Cl) in an inert solvent such =s 1,4-dioxane, N,N- dimethylformamide or THF in the presence of a suitable base su ch as triethylamine or pyridine to generate an appropriate leaving group (compounds of general formula XIV- a-b; LG = Ts or Ms, respectively). Nucleophilic substitution with an appropriate nucleophile (amine or alkoxide) under conditions known to those skilled in the art then gives access to compounds of general formula I-a-b in which R? =R’-(2-4C)alkyl and

R’ is as defined previously.

Conversion of compounds of general formula XI-a-b in wimich A = tert-Bu to carboxylic acids of general formula XII-a-b may be effected b_y deprotection of the tert-butyl ester function. In a typical experiment, the fert-buty] ester of general formula

XI-a-b (A = tert-Bu) is dissolved in dichloromethane and treatezd with a strong acid : such as trifluoroacetic acid. The resulting carboxylic acids of gemneral formula XII-a-b may then be condensed with an appropriate alcohol or amine in the presence of a ' coupling agent such as O-(benzotriazol-1-yl)-N,N,N’,N" -tetramethyluronium tetrafluoroborate (TBTU), O-(7-azabenzotriazol-1-yl)-N,N,N’,N™ -tetramethyluronjum hexafluorophosphate (HATUD) or bromotripyr—rolidinophosphonium hexafluorophosphate (PyBrOP) and a tertiary base, ¢.gz. N,N-diisopropylethylamine, in a solvent such as N,N-dimethylformamide or dichloro-methane at ambient or elevated temperature to give compounds of general formula I-a—b in which R* = R*-carbonyl(1- 4C)alkyl and R® is as defined previously.

NT he

OH ot a ot )n

Mnp® $® N° @ R4 (o} ® hE HN HN hg

Ri —_— @ Ri —— ® R1 vn 8 J J k= A=tBe Ne . 0 . De

A=Me, Et, tBu VID-aR.R*=Me XIaRLR?=Me XII-aR!R? = Me n=1234 VIL-bR!R2=H XFbRL,R*=H XI-bR!,R2=H

Y=1LBr, Cl n=1234 n=1234 ne 0,

OH LG

I 5 ‘) oo” 'n R3

Qo” n o” + § C woo OO oo ¥ x Y

HN — —— HN

C oe C

R1 N 2 Ri

N R2 A N R2 [o}

PN IPN

XIV-aR! R?=Me I-a RI R2=Me

XI-a RI,R2=Me XIV-bRLR2=H IbRIR2=H

XII-bRIR2=H n=1273 : n=123 LG = leaving group

Some of the compounds of the invention, which can be fin the form of a free base, may be isolated from the reaction mixture in the form of a pharmaceutically acceptable salt.

The pharmaceutically acceptable salts may also be obtaimed 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, methanesulphonic acid, fumaric acid, succinic acid, tartaric acid, citric acid, benzoic acid, and ascor®bic acid.

The compounds of the present invention possess at least one chiral carbon atom and may therefore be obtained as pure enantiomers, or ass 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 which are obtaimed from optically active acids and the racemic mixture, or chromatography using chiral columns. For diastereomers, straight phase or reversed phase columns may be used.

The compounds of the invention may form hydrates or solvates. It is known to those of skill in the art that charged compounds form hydrated species when lyophilized with water, or form solvated species when concentrated in a solution with an appropriate organic solvent. The compounds of this invention iraclude the hydrates or solvates of the compounds listed.

For selecting active compounds testing at 10° M must result in an activity of more than 20% of the maximal activity when FSH is used as a meference. Another criterion might be the ECs; value which must be < 10° M, preferably < 107 M.

The skilled artisan will recognize that desirable ECs values are dependent on the compound tested. For example, a compound with an ECs which is less than 10° Mis generally considered a candidate for drug selection. Preferably this value is lower than 107 M. However, a compound which has a higher ECs, but is selective for the particular receptor, may be even a better candidate.

Methods to determine receptor binding, as well ass in vitro and in vivo assays to ‘ determine biological activity, of gonadotropins are well known. In general, the expressed receptor is contacted with the compourad 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 might be

] the Chinese Hamster Ovary cell, but other cells are also suitable. Preferably the cells are of mammalian origin (Jia et al, Mol. Encocrin., 5:759-776, 1991).

Methods to construct 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). Expression of receptor is attained by expression of the DNA encoding the dlesired protein. Techniques for site directed mutagenesis, ligation of additional sequences, PCR, and construction of suitable expression systems are all, by now, well krown in the art. Portions, or all, of the DNA encoding the desired protein can be constncted synthetically using standard solid phase techniques, preferably to include restriction sites for ease of 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 which are compatible with a wide 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 comtacted with the test compound to observe binding, or stimulation or inhibition of a fumctional response.

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

For measurement of binding, radioactively labeled or fluorescently labeled compounds may be used. Also competition binding assays can be performed.

Another assay involves screening for FSH xeceptor 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 exposing the cell to the test compound. The amount of cAMP is ghen measured. The level of cAMP can be reduced or increased, depending on the inhibitory or stimulating effect of the test compound upon binding to the receptor. - Screening for FSH receptor antagonists involves incubation of FSH receptor- expressing cells with a concentration ranges of the test compound in the presence of a fixed, submaximally effective, FSH concentration (i.c., a 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 ICs value and the percentage of inhibition of FSH-inaduced cAMP accumulation can be determined for each of the test compounds. As reference compound human recombinant FSH can be used. In the alternative also competition assays can be performed.

In addition to direct measurement of e. g. cAMP levels in the exposed cell, cells lines can be used which in addition to transfection with receptor encoding DNA are also transfected with a second DNA encocling a reporter gene the expression of which responds to the level of cAMP. Such reporter genes might be cAMP inducible or might be constructed in such a way that they are connected to novel CAMP responsive elements. In general, reporter gene exgpression might be controlled by any response element reacting to changing levels of cAMP. Suitable reporter genes are e.g. the genes encoding P-galactosidase, alkaline phosphatase, firefly luciferase and green fluorescence protein. The principles of such transactivation assays are well known in the art and are described e.g. in Strato-wa, Ch., Himmler, A. and Czernilofsky, A.P., (1995) Curr.Opin.Biotechnol. 6:574.

The present invention also relates to a pharmaceutical composition comprising a tertrahydroquinoline derivative or pharmaceutically acceptable salts thereof having the general formula I in admixture with pharmaceutically acceptable auxiliaries and optionally other therapeutic agents. The auxiliaries must be “acceptable” in the sense of being compatible with the other ingredients of the composition and not deleterious to the recipients thereof.

Compositions include e.g. those switable for oral, - sublingual, subcutaneous, intravenous, intramuscular, local, or reectal administration, and the like, all in unit dosage forms for administration.

For oral administration, the active ingredient may be presented as discrete units, such : as tablets, capsules, powders, granulates., solutions, suspensions, and the like.

For parenteral administration, the pharmaceutical composition of the invention may be presented in unit-dose or multi-dose coxtainers, e.g. injection liquids in predetermined amounts, for example in sealed vials and ampoules, and may also be stored in a ficeeze dried (lyophilized) condition requiring only the addition of sterile liquid carrier, e.g. water, prior to use.

Mixed with such pharmaceutically acceptable auxiliaries, e.g. as described im the standard reference, Gennaro, AR. et al, Remington: The Science and Practice of

Pharmacy (20th Edition., Lippincott Williams & Wilkins, 2000, see especially Paurt 5:

Pharmaceutical Manw facturing), the active agent may be compressed into solid do sage units, such as pills, tablets, or be processed into capsules or suppositories. By means of pharmaceutically acceptable liquids the active agent can be applied as a #fluid composition, e.g. as an injection preparation, in the form of a solution, suspension, emulsion, or as a sprayy, e.g. a nasal spray.

For making solid dosage units, the use of conventional additives such as filJers, colorants, polymeric binders and the like is contemplated. In general any phar-ma- ceutically acceptable additive which does not interfere with the function of the active compounds can be used. Suitable carriers with which the active agent of the invention can be administered as solid compositions include lactose, starch, cellulose derivat ives and the like, or nmixtures thereof, used in suitable amounts. For parenteral administration, aqueows suspensions, isotonic saline solutions and sterile injectable solutions may be used, containing pharmaceutically acceptable dispersing ageents and/or wetting agents, such as propylene glycol or butylene glycol.

The . invention further includes a pharmaceutical composition, as hereinbefore described, in combination with packaging material suitable for said composition, said packaging material including instructions for the use of the composition for the uses as hereinbefore described.

The tetrahydroquinolirze derivatives of the invention can also be administered in the form of implantable pharmaceutical devices, consisting of a core of active matenrial, encased by a release rate-regulating membrane. Such implants are to be applied subcutaneously or locally, and will release the active ingredient at an approximautely constant rate over relatively large periods of time, for instance from weeks to ye ars.

Methods for the preparation of implantable pharmaceutical devices as such axe known in the art, for exarmple 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 cosmposition thereof, will necessarily be dependent upon the therapeutic effect to be achiesved (treatment of infertility; contraception), and may vary= with the particular compound, the route of administration, and the age and condition of the individual subject to whom the medicament is to be administered.

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

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

A further aspect oef the invention resides in the use of a tetrahydroquinoline clerivative compound having: the general formula I for the manufacture of a medicament to be used for the treatument of disorders responsive to FSH receptor mediated pathways.

Thus, patients in need thereof can be administered with suitable amoun®#s of the compounds accorcling to the invention.

In another aspect the invention resides in the use of a tetrahydroguinoline clerivative compound having: the general formula I for the manufacture of a medicameent to be used for the control of fertility.

In yet another aspect the invention resides in the use of a tetrahydroquinoline Qerivative } compound having the general formula I for the manufacture of a medicament to be used for the treatmment of infertility.

In still another aspect the invention resides in the use of a -tetrahydroquinoline dermvative compound having the general formula I for the manufacture of a meciijcament to be used to prevent fertility.

The compounds according to the invention can also be used for the treatment of horamone-dependent disorders such as breast cancer, prostate cancer and endometriosis.

The invention is illustrated by the following examples.

Examples

Gemeral Comments

The following abbreviations are used in the examples: DMA = NN, N-dimethylaniline,

DIPEA = N,N-diisopropylethylamine; TFA = trifluoroacetic acid, DIBAD = di-ter- butyl azodicarboxylate; TBTU = O-Benzotriazole-1-yl-N,N,N’,N ’-stetramethyluronium tetraafluoroborate; HATU = O-(7-azabenzotriazole- 1-yl)-N, N,N’, N -#tetramethyluronium : hex afluorophosphate; Fmoc = 9-fluorenylmethoxycarbonyl; Fmoc-Cl = 9- fluorenylmethoxycarbonylchloride; DMF = N,N-dimethylformamnide; Boc = tert- butoxycarbonyl; THF = tetrahydrofuran.

The names of the final products described in the examples are generated using the

Beilstein Autonom program (version: 2.02.119).

Unless stated otherwise, all final products of the examples below axe lyophilized from water/1,4-dioxane mixtures or water/acetonitrile mixtures. If the compound was prepared as a HCI- or TFA sal, the respective acids were added in amppropriate amounts to thhe solvent mixture before lyophilization.

Thes following analytical HPLC methods are used for determination of retention times:

Method 1: Column: 5 pm Luna C-18(2) 150%4.6 mm; flow: 1ml/mnin; detection: 210 nmg column temperature: 40 °C; solvent A: CH;CN/H20 = 1/9 (v/v); solvent B:

CH=CN; solvent C: 0.1 M aqueous trifluoroacetic acid; gradient: solvent A/B/C = 65/30/5 to 10/85/5 (v/v/v) in 30.00 min, then constant for an addi-tional 10.00 min at

A/B8/C =10/85/5 (vIVIv). ) Method 2: Identical to method 1, except for the gradient used: Grad_ient: solvent A/B/C = 75/20/5 to 15/80/5 (v/v/v) in 30.00 min, then constant for an additional 10.00 min at

A/BR/C = 15/80/5 (vIviv).

Method 3: Column: 3 ym Luna C-18(2) 100x2 mm; flow: 0.25 ml/min; detectiom: 210 nm; column temperatuare: 40 °C; solvent A: H,O; solvent B: CH;CN; solvent C: 5 0 mM phosphate buffer, pH 2.1; gradient: solvent A/B/C = 70/20/10 to 10/80/10 (v/x2/v) in 20.00 min, then constawt for an additional 10.00 min at A/B/C = 10/80/10 (v/v/v).

Method 4: Identical to» method 3, except for the gradient used: Gradient: solvent .A/B/C = 65/30/5 to 10/85/5 Cv/v/v) in 20.00 min, then constant for an additional 10.00 amin at

A/B/C = 10/85/5 (v/v/v).

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

Method 6: Identical tos method 1, except for the gradient used: Gradient: solvent .A/B/C = 35/60/5 to 10/85/5 (v/v/v) in 30.00 min, then constant for an additional 10.00 mmin at

A/B/C =10/85/5 (v/v/v).

The following methods are used for preparative HPLC-purifications:

Method A: Column == Luna C-18. Gradient: 0.1% trifluoroacetic acid in H,O/CH;CN (9/1, viW)/ICH3CN = 100/0 to 0/100 (v/v) in 30-45 min, depending on the ease of separation. Detectiom: 210 nm. The appropriate fractions were collected and concentrated (partially=) in vacuo.

Method B: Column = Luna C-18. Gradient: H,O/CH5CN (9/1, v/v)/CH;CN = 83/20 to 0/100 (v/v) in 30-45 min, depending on the ease of separation. Detection: 210 nna.

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-Trimethyl- A .2-dihydroquinolin-6-yl)-carbamic acid terz-butyl ester

A mixture of N-B oc-1,4-phenylenediamine (75 g), MgSO: (216 g), <4-tert- butylcathechol (1.8 g)) and iodine (4.7 g) in anhydrous acetone (600 ml) was Theated under reflux for 20 kn. The MgSO 4 was removed by filtration and the filtrate was concentrated in vacuoe. The residue was chromatographed on a short plug of sillicagel using heptane/ethyl acetate = 8/2 (v/v) as the eluent to give the product as a browm oil. : 30 Yield: 41 g.

) (b). (1-Acetyl-2.2 4-trimethyl-1,2-dihydroquinolin-6-y1)-carbamic acid tert-butyl ester

As solution of the compound described in example 1a (41 g) in pyridine (200 ml) and

CHCl; (200 ml) was cooled to 0 °C. Acetyl chloride (21 ml) in CHCl, (50 ml) was added dropwise. After complete addition the mixture was stirced for 3 h at room temperature. Ethyl acetate (2 I) and H2O (2 1) were added and thhe organic layer was separated, dried and concentrated in vacuo. The title compouand was obtained by cxystallization from ethyl acetate.

Yield: 23 g. (©). 1-Acetyl-6-amino-2,2.4-trimethyl-1,2-dihydroguinoline

T he compound described in example 1b (15 g) was stirred in a moixture of CHCl; and

TFA (9/1 (v/v), 300 ml) for 2 h. The reaction mixture was cooled own to 0 °C, and the pH adjusted to pH 7 using a 2 M aqueous NaOH solution. Tize organic layer was separated, washed with brine, dried and concentrated in vacueo to give the crude "15 pwoduct that was used without further purification in the next step.

Yield: 104 g (AQ). Biphenyl-4-carboxylic acid (1-acetyl-2,2,4-trimethyl-1,2-d&hydroquinolin-6-yl)- amide

T-0 a solution of the compound described in example 1c (10 g) aad DIPEA (40 ml) in

CHCl (100 ml), was added 4-biphenylcarbonyl chloride (9.8 g) and the resulting mixture was stirred for 18h at room temperature. Water was addeed, the organic layer was separated, dried and concentrated in vacuo. The product w~as crystallized from ethyl acetate.

Yield:15g (e) Biphenyl-4-carboxylic acid [1-acetyl-4-(4-methoxyphenyl)-2.2 4-trimethyl-1.2,3.4- testrahydroquinolin-6-yl]-amide

While stirring, aluminum trichloride (9.7 g) was added to a mixtire of the compound } 30 described in example 1d (10.0 g) and anhydrous anisole (50 mal) and the resulting mixture was stirred at 35 °C for 18 h. After this time, water was added at 0 °C and the re=sulting 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 h.

“The formed precipitate was collected by filtration and dried in vacuo to give the title ) «compound. “Yield: 79 g. &f). Biphenyl-4-carboxylic acid [1-acetyl-4-(4-hydroxyphemyl)-2.2,4-trimethyl-1.2,3.4-

Eetrahydro-quinglin-6-yl)-amide "To a solution of the compound described in example 1e (7.9 g) in CHCl, (200 ml) at 0 “C was added a solution of boron tribromide (5 ml) in CH, Cl, (50 ml) and the mixture wvas kept for 4 h at 0 °C. Water (ca 500 ml) was carefully added and the resulting rmixture was vigorously stirred. The organic layer wvas separated, dried and

Concentrated in vacuo. Crystallization from ethyl acetate afforded the title compound.

Wield: 6.1 g. (8) 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

General procedure A: To a solution of the compound described in example 1f (70 mng) in DMF (2 ml) were added Cs2CO; (200 mg) and 2-diimethylamino-ethylchloride hydrochloride (17 mg). The resulting mixture was stirred owvernight, after which water amd ethyl acetate were added. The organic layer was separated, dried and concentrated ix vacuo. The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH;CN and water containing TFA to give the corresponding TFA salt.

Yield: 18 mg (TFA salt); MS-ESL: [M+H]" = 576.6; HPLC: R,= 14.96 min (method 3).

E-xample 2

Beiphenyl-4-carboxylic acid {l-acetyl-4-[4-(2-dimethylamiz1o-propoxy)-phenyl}-2.2,4- trimethyl-1,2 3 4-tetrahydro-quinolin-6-yl}-amide

A_ccording to general procedure A, the compound described in example 1f (70 mg) was akkylated with 3-dimethylamino-propylchloride hydrochloride (19 mg) and Cs.COs i 30 (200 mg) in DMF (2 ml). The product was purified by preparative HPLC (method A) ard lyophilized from a mixture of CH;CN and water containing TFA to give the corresponding TFA salt.

Yield: 58 mg (TFA salt); MS-ESI: [M+H]" = 590.48; HPLC: R; = 15.36 min (method 3).

Example 3

Biphenyl-4-carboxylic acid {1-acetyl-2.2 _4-trimethyl-4-[4-(3-morpholin-4-yl- propoxy)-phenyl]-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-morpholinopropylchloride (26 mgz) and CsCO3 (200 mg) in DMF (2 ml). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH;CN and water containing TFA to giwe the corresponding TFA salt.

Yield: 56 mg (TFA salt); MS-ESI: [M+H]" = 631.6 ; HPLC: R, = 15.40 min (method 3).

Example 4

Biphenyl-4-carboxylic acid _ {1-acetyl-2.2.4-tri methyl-4-[4-(pyridin-2-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 2-picolyl chloride hydrochlorid_e (33 mg) and Cs;COj; (325 mg) in

DMF (5 ml). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CHsCN and water containing TFA to give the corresponding TFA salt.

Yield: 60 mg (TFA salt); MS-ESI: [M+H]" = 596.45 HPLC: R; = 19.75 min (method 2).

Examples

Biphenyl-4-carboxylic acid {1-acetyl-2.2 4-tr—imethyl-4-[4-(1-methyl-piperidin-3- ylmethoxy)-phenyl]-1,2,3.4-tetrahydro-quinolin-6-w»1}-amide

According to general procedure A, the compound described in example if (100 mg) was alkylated with 3-chloromethyl-1-methylpiper-idine hydrochloride (33 mg) and

Cs2CO; (325 mg) in DMF (5 ml). The product wwas purified by preparative HPLC (method A) and lyophilized from a mixture of CHI;CN and water containing TFA to ’ give the corresponding TFA salt.

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

Example 6

Biphenyl-4-carboxylic acid {1-a.cetyl-4-[4-(2-diethylamino-ethoxy)-phenyl]-2.2.4- trimethyl-1,2.3,4-tetrahydro-quinolin—6-yI}-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,CO3 (325 mg) in DMF (5 ml). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CHsCN and water containing TFA to give the corresponding TFA salt.

Yield: 67 mg (TFA salt); MS-ESI: [MI+H]" = 604.4; HPLC: R, = 16.38 min (method 2).

Example 7

Biphenyl-4-carboxylic acid __{1-aacetyl-2.2.4-trimethyl-4-[4-(pyridin-4-ylmethoxy)- phenyl]-1,2.3 4-tetrahydro-quinolin-&-yl} -amide

According to general procedure A, the compound described in example 1f (100 mg) was alkylated with 4-picolylchloride hydrochloride (33 mg) and Cs,CO3 (325 mg) in

DMF (5 ml). The product was pawified by preparative HPLC (method A) and lyophilized from a mixture of CH3;CN and water containing TFA to give the corresponding TFA salt.

Yield: 61 mg (TFA salt); MS-ESI: [MI+H]" = 596.4; HPLC: R, = 16.64 min (method 2).

Example 8

Morpholine-4-carboxylic acid [3-(4—{1-acetyl-6-[(biphenyl-4-carbonyl)-amino}-2.2.4- trimethyl-1.2 3 4-tetrahydro-quinolin—4-yl1}-phenoxy)-propyl]-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

Cs2COs (325 mg) in DMF (5 ml). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH3CN and water.

Yield: 95 mg; MS-ESIL: [M+H]" = 675.6; HPLC: R, = 18.24 min (method 3).

Example 9

Biphenyl-4-carboxylic acid __{1-~acetyl-4-[4-(2-azepan- 1-yl-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-(hexamethyleneimino)ethyl chloride hydrochloride (42 mg) and

Cs:CO5 (325 mg) in DMF (5 ml). The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH;CN and water containing TFA to give the corresponding TFA salt.

Yield: 60 mg (TFA salt); MS-ESI: [M-£H]" = 630.6; HPLC: R, = 17.25 min (method 2).

Example 10

Bipheny}-4-carboxylic acid _{1-ac etyl-2.2 4-trimethyl-4-[4-(pyridin-3-ylmethoxy)- phenyl]-1,2,3 4-tetrahydro-quinolin-6-w/1} -amide

According to general procedure A, the- compound described in example 1f (1.0 g) was alkylated with 3-picoloylchloride hydrochloride (488 mg) and Cs,CO; (3.2 mg) in

DMF (10 ml). The product was puwified by preparative HPLC (method A) and lyophilized from a mixture of CHxzCN and water containing TFA to give the corresponding TFA salt.

Yield: 884 mg (TFA salt); MS-ESI: [N+H]" = 596.4; HPLC: R, = 16.55 min (method 3).

Example 11

Biphenyl-4-carboxylic acid [1-acetyl-4--(4-carbamoylmethoxy-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-chloroacetamide (24 mg) and Cs>CO; (325 mg) in DMF (5 ml).

The product was purified by preparative HPLC (method A) and lyophilized from a mixture of CH3CN and water containing TFA to give the corresponding TFA salt.

Yield: 40 mg; MS-ESL: [M+H]" = 562.66; HPLC: R; = 21.63 min (method 2).

Example 12

Biphenyl-4-carboxylic acid _ [1-aceetyl-4-(4-allylcarbamoylmethoxy-phenyl)-2.2.4- trimethyl-1.2.3 4-tetrahydro-quinolin-6—yl]-amide (a). (4-{1-Acetyl-6-[(biphenyl-4-cartyonyl)amino]-2,2.4-trimethyl-1.2,3 4-tetrahydro- ' quinolin-4-yl}phenoxy)acetic acid tert-butyl ester

A mixture of the compound described -in example 1f (2.58 g), tert-butyl bromoacetate ) (826 ul), K2COs (2.8 g) and acetone (1900 ml) was stirred for 18 h at 50 °C. The solids were removed by filtration and the filirate was concentrated in vacuo to give the product that was used without further purification in the next step.

. Yield:32g (b). (4-{1-Acetyl-6-[(lbiphenyl-4-carbonyl)amino]-2.2 4-trimethyl-1,2.3 4-tetrahydro- quinolin-4-yl}phenoxy)acetic acid

The compound described in example 12a (3.2 g) was stirred in a mixture of CH,Cl, amd

TFA (9/1 (v/v), 100 mH) for 3 h. Toluene (100 ml) was added and the mixture w-as concentrated in vacuo to give the crude product, that was used without furthmer purification.

Yield:33 g (c). Biphenyl-4-carthoxwlic acid [1-acetyl-4-(4-allylcarbamoylmethoxy-phenyl)-2.2.4- trimethyl-1,2.3.4-tetrahyzdro-quinolin-6-yl]-amide

General procedure B: To a solution of the compound described in example 12b (82 mg), allylamine (37 mg) and DIPEA (226 pl) in CHCl; (5 ml) was added TBTU (84 mg) at room temperatuwe. 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: fM+H]" = 602.4; HPLC: R; = 18.19 min (method 4).

Example 13

Biphenyl -4-carboxylic acid __{1-acetyl-4-[4-(isopropylcarbamoyl-mcthoxy)-pheny3]- 2,2 A-trimethyl-1,2,3 4-tetrahydro-quinolin-6-yl}-amide

According to general procedure B, the compound described in example 12b (82 mxg) was treated with isopropylamine (38 mg), DIPEA (226 pl) and TBTU (84 mg) in

CHCl (5 ml). The title compound was purified by preparative HPLC (method A).

Yield: 45 mg; MS-ESI: [M+H]" = 604.6; HPLC: R, = 18.63 min (method 4).

Example 14 ) Biphenyl-4-carboxylic _ acid [1-acetyl-4-(4-diethylcarbamoylmethoxy-phenyl)-2.2, 4- trimethyl-1,2,3.4-tetrahy-dro-quinolin-6-yl]-amide

According to general procedure B, the compound described in example 12b (82 m.g) was treated with diethylamine hydrochloride (47 mg), DIPEA (226 pl) and TBTU (84 mg) in CHCl; (5 ml). The title compound was purified by preparative HPLC (method

A).

Yield: 51 mg; MS-ESSI: [M+H]" = 618.4; HPLC: R, = 19.09 min (method 4).

Example 15

Biphenyl-4-carboxyl#sc acid [l-acetyl-2.2 4-trimethyl-4-(4- {[(pyridin-4-ylmetheyl)- catbamoyl]-methoxy }-phenyl)-1,2,3.4-tetrahydro-quinolin-6-yi}-amide

According to general procedure B, the compound described in example 12b (82 mmg) was treated with 4-poicolylamine (70 mg), DIPEA (226 pl) and TBTU (84 mg) in

CHCl; (5 ml). The title compound was purified by preparative HPLC (method A) and lyophilized from a mixture of CH;CN and water containing TFA to give the corresponding TFA s=alt.

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

Example 16

Biphenyl-4-carboxyliic acid [1-acetyl-4-(4-{[(furan-2-ylmethyl)-carbamoyl]-methoxy}- phenyl}-2.2 4-trimetlmyl-1.2.3 4-tetrahydro-quinolin-6-yl]-amide

According to general procedure B, the compound described in example 12b (82 mmg) was treated with 2-Furfurylamine (63 mg), DIPEA (226 ul) and TBTU (84 mg) in

CH.C; (5 ml). The title compound was purified by preparative HPLC (method A).

Yield: 50 mg; MS-ESI: [M+H]" = 642.6; HPLC: R;= 21.31 min (method 3).

Example 17

Biphenyl-4-carboxyliic_ acid (1-acetyl-4-{4-[(2-methoxy-ethylcarbamoyl)-metho><y]- phenyl}-2.2 4.trimethayl-1,2,3 4-tetrahydro-quinolin-6-yl)-amide

According to generall procedure B, the compound described in example 12b (82 mmg) was treated with 2-meethoxyethylamine (49 mg), DIPEA (226 ul) and TBTU (84 mes) in

CHCl; (5 ml). The title compound was purified by preparative HPLC (method A). : Yield: 34 mg; MS-ESSI: [M+H]" = 620.4; HPLC: R; = 19.70 min (method 3).

. Example 18

Biphemyl-4-carboxylic acid {1-acetyl-4-[4-(benzylcarbamoyl-methhoxy)-phenyl}-2,2.4- trimetthyl-1,2.3.4-tetrahydro-quinolin-6-yl} -amide

According to general procedure B, the compound described in e>xample 12b (82 mg) was treated with benzylamine (49 mg), DIPEA (226 pl) and TBTUJ (84 mg) in CH,CL (5 ml}. The title compound was purified by preparative HPLC (method A).

Yield: 53 mg; MS-ESI: [M+H]" = 652.6; HPLC: R, = 22.26 min (method 3).

Exam ple 19

Biphemyl-4 carboxylic acid __ (1-acetyl-4- {4-[(2-dimethylamimo-ethylcarbamoyl)- methoxy]-phenyl}-2.2 4-trimethyl-1,2.3.4-tetrahydro-quinolin-6-yl )-amide

According to general procedure B, the compound described in exxample 12b (82 mg) was treated with N,N-dimethylethylenediamine (49 mg), DIPEA «(226 pl) and TBTU (84 mg) in CHCl, (S ml). The title compound was purified by preparative HPLC (method A). Lyophilization from a mixture of aqueous HCI and E ,4-dioxane afforded the titLe compound as a HCl-salt.

Yield: 11 mg (HCl-salt); MS-ESI: [M+H]* = 633.4; HPLC: R,= 13 .74 min (method 3).

Example 20

Biphemyl-4-carboxylic acid [1-acetyl-2.2 4-trimethyl-4-(4-methyk carbamoylmethoxy- pheny®)-1.2.3 4-tetrahydro-quinolin-6-yl}-amide

Accorcling to general procedure B, the compound described in exzample 12b (82 mg) was treated with methylamine hydrochloride (20 mg), DIPEA (226 pl) and TBTU (84 mg) in. CHCl, (5 ml). The title compound was purified by preparative HPLC (method

A).

Yield: 35 mg; MS-ESI: [M+H]" = 576.4; HPLC: R; = 19.25 min (nmethod 3).

Exampple 21 : . Bipheryyl-4-carboxylic acid {1-acetyl-2.2 4-trimethyl-4-[4-(2-morpholin-4-yl-2-oxo- ethoxy-)-phenyl]-1.2 3 4-tetrahydro-quinolin-6-yl}-amide . Accorcling to general procedure B, the compound described in example 12b (110 mg) was treated with morpholine (74 mg), DIPEA (296 pl) and TBTU «109 mg) in CHCl, (5 mi). The title compound was purified by preparative HPLC (metlnod A).

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

Example 22

N-{1-Acetyl-2 2 ,4-trimethyl-4-[4-(3-morpholin-4-yl-preopoxy)-phenyl]-1,2.3.4- tetrahydro-guinolin-6-yl1}-5-bromo-2-methylamino-ben zamide (a). (1-Acety}-2.2 4-trimethyl-1.2-dihydro-quinolin-6—yl)-carbamic acid _9-fluoren- ylmethyl ester

To a solution of the compound described in example Lc (17 g) and DIPEA (40 ml) in

CHCl, (100 ml), was added FmocCl (25 g) and the resulting mixture was stirred for 18h 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 silicagel using CH;ClL, as the eluent.

Yield: 16.6 g (b) [1-Acetyl-4-(4-methoxyphenyl)-2.2 4-trimethyl-T 2.3 4-tetrahydroquinolin-6-yl]- carbamic acid 9-fluorenylmethyl ester

While stirring, aluminum trichloride (24.2 g) was addead 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 h. After this time, vwvater was added at 0 °C and the resulting mixture was extracted with ethyl acetate. Thhe organic layer was separated, dried and partially concentrated in vacuo and the mixture was stored at 0 °C for 18 h.

The formed precipitate was collected by filtration and dried in vacuo to give the title compound.

Yield: 10.1 g. (¢©) [1-Acetyl-4-(4-hydroxyphenyl)-2.2.4-trimethyl-1 2.3 4-tetrahydroquinelin-6-yi}- carbamic acid 9-fluorenylmethyl ester

To a mixture of the compound described in example 227d (10.1 g) in anhydrous CH,Ch (500 ml) was added dropwise boron tribromide (5.05 mH) and the resulting mixture was } stirred for 2.5 h at room temperature. The reaction was quenched with ice water at 0 °C and CHCl; was added. The organic layer was separated, dried and stored at 4 °C for 20 30h. The formed solids were collected by filtration and d_ried in vacuo to give the crude product that was used without further purification.

Yield: 12.5 g.

(d). 1-Acetyl-6-amino-2,2 4-trimethyl-4-[4-(3-naorpholin-4-yl-propoxy)-phenyi]- 1.2.3 4-tetrabydroquinoline

A mixture of the compound described in example 22-¢ (1.0 g), Cs2COs (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,Cl,. The organic layer was dried and concentrated in vacuo. The title compound was purified by chromatography on silicagel using CH>Cly/ 2% concentrated ammonia in MeOH = 1/0 => 9/1 (v/v) as the eluent.

Yield 527 mg (e). N-{1-Acetyl-2.2 4-trimethy}-4-[4-(3-morpholir-4-yl-propoxy)-phenyl]-1,2.3 4- tetrahydro-quinolin-6-yl}-5-bromo-2-methylamino-ben zamide

General procedure C: To a solution of the compound described in example 22d (132 mg), 5-bromo-2-methylamino benzoic acid (101 mg) amd DIPEA (255 pl) in CHCl (3 ml) was added HATU (166 mg) at room temperature. "The reaction mixture was stirred for 18 h at room temperature. Ethyl acetate (15 ml) arid 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-ESL [M+H]" = 663.4; HPLC: R, = 14.65 min (method 3).

Example 23

N-{1-Acetyl-2 2 4-trimethyl-4-[4-(3-morpholin-4-yl-propoxy)-phenyl]-1.2,3.4- tetrahydro-quinolin-6-yl}-3,5-dichloro-2.6-dimethoxy-bwenzamide

According to general procedure C, the compound described in example 22d (132 mg) was acylated with 3,5-dichloro-2,6-dimethoxybenzoic axcid (110 mg), DIPEA (255 pl) and HATU (166 mg) in CH>Cl; (3 ml). The title compoand was purified by preparative

HPLC (method A). . Yield: 68.3 mg; MS-ESI: [M+H]* = 684.3; HPLC: R, = 13.45 min (method 3). . Example 24

Biphenyl-4-carboxylic acid _ [1-acetyl-4-(4-{2-[(furam-2-ylmethyl)-amino]-ethoxy}- henyl)-2.2,4-trimethyl-1,2.3.4-tetrahydro-quinolin-6-v1 ]-amide i -33- (a). (4-{-Acetyl-6-[(biphenyl-4-carbonylamino}-2.2,4-trimethyl-1.2 .3 4-tetrahydro- quinolin-4-yl} -phenoxy)acetic acid ethyl ester

A mixture «of the compound described in example 1f(1 g), ethyl bromoa cetate (220 pl),

K2CO; (850 mg) and acetone (25 ml) was stirred for 6 h at 50 °C. The solids were removed by filtration and the filtrate was concentrated in vacuo to give the product that was used without further purification in the next step.

Yield: 1.2 g (b). Biphleenyl-4-carboxylic acid _ {1-acetyl-4-[4-(2-hydroxyethoxy )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 carefullly added LiALH, (78 mg), and the resulting mixture was stirred for 3 h at room temperature. Ethyl acetate (50 ml) was added dropwise, followed by water (50 ml). The acqueous 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 concentrate=d in vacuo to give the product that was used without further purification in the next step. ‘

Yield: 1 g (c). Metlaagesulfonic acid 2-(4-{1-acetyl-6-{(biphenyl-4-carbonyB)amino}-2.2 4- trimethyi-1.,2.3 4-tetrahydroquinolin-4-yl} phenoxy)ethyl ester

To a solution of the compound described in example 24b (1 g) and DIPEA (1.7 ml) in

CHCl; (15 ml), was added dropwise a solution of methanesulfonyl chloxride (310 pl) in

CHC (5 mi). After 2 h, water was added, the organic layer separated, dried and concentrated in vacuo. The title compound was purified by chromnatography on silicagel using heptane/ethyl acetate = 9/1 => 1/1 (v/v) as the eluent.

Yield: 870 mg (d). Biphen-yl-4-carboxylic acid [1-acetyl-4-(4-{2-[(furan-2-ylmethyl)-amnino]-ethoxy}- . 30 phenyl)-2.2 4-trimethyl-1,2,3 4-tetrahydro-quinolin-6-yl]-amide

General pr-ocedure D: To a solution of the compound described in example 24c (87 mg) in CH3 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 amd the product

] was purified by preparative HPLC (method A) and lyophilized from = mixture of

CH;CN amd water containing TFA to give the corresponding TFA salt.

Yield: 47 mg (TFA salt); MS-ESI: [M+H]" = 628.6; HPLC: R, = 11.53 mina (method 4).

Example 25

Biphenyl—4-carboxylic acid (1-acetyl-4-{4-[2-(2-hydroxy-1,1-dimethyl-esthylamino)- ethoxy]-pehenyl}-2.2 4-trimethyl-1,2,3 4-tetrahydro-quinolin-6-yl)-amide

Accordin_g to general procedure D, the compound described in example 24c (87 mg) was treateed with 2-amino-2-methyl-propan-1-ol (100 mg) in CH3;CN (5 mml). The title compound was purified by preparative HPLC (method A) and lyophil ized from a mixture oof CH3CN and water containing TFA to give the corresponding TF= A salt.

Yield: 21 mg (TFA salt); MS-ESI: [M+H]" = 619.8; HPLC: R; = 10.95 mim (method 4).

Example= 26

Biphenyl—4-carboxylic acid _[1-acetyl-2,2 4-trimethyl-4-(4-{2-[(pyridin- 3-ylmethyl)- amino]-esthoxy}-phenyl)-1.2,3,4-tetrahydro-quinolin-6-yl]-amide

Accordin g to general procedure D, the compound described in example =24c (87 mg) was treasted with 3-aminomethylpyridine (119 mg) in CHaCN (5 mM). The title : compoun~d was purified by preparative HPLC (method A) and lyophilized from a mixture ov CH3;CN and water containing TFA to give the corresponding TFA salt.

Yield: 40 mg (TFA salt); MS-ESI: [M+H]" = 639.4; HPLC: R; = 10.15 mim (method 4).

Examples 27

Biphenyl—4-carboxylic____ acid 1-acetyl-4- {4-[2-(2-hydroxy-ethylami no)-ethoxy]- henyl}-=2.2 4-trimethyl-1,2,3 4-tetrahydro-quinolin-6-yl)-amide

Accordin_g to general procedure D, the compound described in example 224¢ (100 mg) was treated with ethanolamine (100 mg) in CH3CN (5 ml). The title compound was purified by preparative HPLC (method A) and lyophilized from a mixture of CH;CN and watemr containing TFA to give the corresponding TFA salt

Yield: 50 mg (TFA salt); MS-ESI: [M+H]" = 592.6; HPLC: R, = 10.32 mir (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-tetrahydro-quinolin-6-yl)-amide

According to general parocedure D, the compound described in example 24c (100 mg) was treated with ethylemediamine (110 mg) in CH3CN (5 ml). The title compound was purified by preparative HPLC (method A) and lyophilized from a mixture of CIH;CN and water containing TIFA to give the corresponding TFA salt

Yield: 45 mg (TFA salt); MS-ESIL: [M+H]* = 591.4; HPLC: R, = 7.04 min (method 1).

Example 29

Biphenyl-4-carboxylic acid {1-acetyl-2.2 4-trimethyl-4-[4-(2-piperazin-1-yl-eth oxy)- phenyl}-1.2.3 4-tetrahyro-quinolin-6-yl} -amide

According to general procedure D, the compound described in example 24¢ (100 mg) was treated with piperazine (140 mg) in CH;CN (5 mi). The title compouncl was purified by preparative HPLC (method A) and lyophilized from a mixture of CEL;CN and water containing TIFA to give the corresponding TFA salt

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

Example 30

Morpholine-4-carboxyl-ic acid (3-{4-[1-acetyl-6-(3.5-dichloro-2,6-dimetthoxy- benzoylamino)-2.2.4-triimethyl-1,2 3 4-tetrahydro-quinolin-4-yl]-phenoxy} -propy¥)- amide (a). Morpholine-4-carboxylic acid (3-{4-[1-acetyl-6-amino-2,2,4-trimethyl-1,2.3.4- tetrahydro-quinolin-4-y~1]-phenoxy}-propyl)-amide

According to the same procedure described in example 22d, the compound described in example 22c (1.0 g), wras alkylated (with concomitant removal of the Fmoc protective group) with morpholime-4-carboxylic acid (3-chloropropyl)amide (448 mg) using

Cs;CO;3 (1.8 g) in DMIF (5 ml). The title compound was purified by chromatography on silicagel using CH>CL/~ 2% concentrated ammonia in MeOH = 1/0 => 9/1 (v/v) as the eluent. . Yield: 894 mg.

) (b). Morpholine-4-camboxylic acid _(3-{4-{1-acetyl-6-(3.5-dichloro-2.6-dimethoxy- benzoylamino)-2.2.4-tr-imethyl-1,2,3.4-tetrahydro-guinolin-4-yl]-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 pl) and HATU (609 mg) im CH,Cl; (5 ml). The title compound was purified by preparative

HPLC (method A).

Yield: 102 mg; MS-ES T: [M+H]" = 727.4; HPLC: R; = 22.37 min (method 2).

Example 31

N-{1-Acetyl-2.2 4-trimmethyl-4-[4-(2-morpholin-4-yl-ethoxy)-phenyll-1.2.3.4- tetrahydro-quinolin-6-ys1}-3.5-dibromo-benzamide (a). 1-Acetyl-6-amino—2.2 4-trimethyl-4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-1.2.3.4- tetrahydroquinoline

A mixture of the compound described in example 22¢ (1.0 g), Cs:CO; (1.8 g), N-(2- chloroethyl)-morpholirae hydrochloride (375 mg), and DMF (5 ml) was stirred at 60 °C for 18 h. The reaction «did not reach completion and additional amounts of Cs;CO; and

N-(2-chloroethyl)-morpholine hydrochloride were added. After the reaction was complete, water was a_dded and the mixture was extracted with CH,Cl;. The organic layer was dried and concentrated in vacuo. The title compound was purified by chromatography on silimcagel using CH,Cly/ 2% concentrated ammonia in MeOH = 1/0 => 9/1 (v/v) as the elue=nt.

Yield: 905 mg. (b). N-{1-Acetyl—2.2 4-trimethyl-4-[4-(2-morpholin-4-yl-ethoxy)-phenyl}-1.2.3 4- tetrahydro-quinolin-6-y/1}-3,5-dibromo-benzamide

According to general procedure C, the compound described in example 31a (157 mg) was acylated with 3,5--dibrombenzoic acid (150 mg), DIPEA (313 ul) and HATU (204 , mg) in CH2Cl; (5 ml). The title compound was purified by preparative HPLC (method

A). : 30 Yield: 71 mg (TFA sals); MS-ESI: [M+H]" = 700.2; HPLC: R; = 16.12 min (method 2).

) Example 32

N-{1-Acetyl-2.2 4-trimethyl-4-[4-(2-meorpholin-4-yl-ethoxy)-phenyl]-1.2,3.4- tetrahydro-quinolin-6-y1}-2-chloro-bem zamide

According to general procedure C, the: compound described in example 31a (150 mg) was acylated with 2-chlorobenzoic acicl (81 mg), DIPEA (299 pul) and HATU (195 mg) in CHCl, (6 ml). The title compound wwvas purified by preparative HPLC (method A).

Yield: 162 mg (TFA salt); MS-ESI: [ME+H]" = 576.4; HPLC: R, = 9.37 min (method 2).

Example 33

N-{1-Acetyl-2.2 4-trimethyl-4-[4-(2-meorpholin-4-yl-ethoxy)-phenyl]-1,2.3 4- tetrahydro-quinolin-6-yl1}-3.5-dimethyl ~benzamide

According to general procedure C, the- compound described in example 31a (200 mg) was acylated with 3,5-dimethylbenzo®c acid (103 mg), DIPEA (399 pl) and HATU (260 mg) in CHCl, (7.5 ml). The title compound was purified by preparative HPLC (method A).

Yield: 57.5 mg (TFA salt); MS-ESI: [W+H]' = 570.4; HPLC: R; = 12.62 min (method 2).

Example 34

N-{1-Acetyl-2 2 4-trimethyl-4-[4-(2-merpholin-4-yl-ethoxy)-phenyl]-1.2.3.4- tetrahydro-quinolin-6-yl}-2 5-dichloro—~benzamide

According to general procedure C, the compound described in example 31a (200 mg) was acylated with 2,5-dichlorobenzoic acid (131 mg), DIPEA (399 ut) and HATU (260 mg) in CH;Cl (7.5 ml). The title compound was purified by preparative HPLC (method A).

Yield: 130 mg (TFA salt); MS-ESI: [N+H]" = 610.2; HPLC: R, = 11.70 min (method 2).

Example 35 } N-{1-Acetyl-2 2 4-trimethyl-4-[4-(2-morpholin-4-yl-ethoxy)-phenyl}-1.2.3 4- tetrahydro-quinolin-6-yl1}-5-methyl-2-mitro-benzamide

According to general procedure C, the: compound described in example 31a (157 mg) was acylated with 5-methyl-2-nitroben=zoic acid (97.3 mg), DIPEA (313 pl) and HATU

. (204 mg) in CHCl, (5 ml). The title compound was purified by preparative HPLC {method A).

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

Example 36

N-{1-Acetyl-2.2 4-trimethyl-4-[4-(2-moOrpholin-4-yl-ethoxy)-phenyl}-1.2.3.4- tetrahydro-quinolin-6-yl}-benzamide

According to general procedure C, the compound described in example 31a (157 mg) was acylated with benzoic acid (65.6 mg), DIPEA (313 pl) and HATU (204 mg) in

CHCl, (5 ml). The title compound wass purified by preparative HPLC (method 4).

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

Example 37

N-{1-Acetyl-2.2 4-trimethyl-4-[4-(2-merpholin-4-yl-ethoxy)-phenyl]-1.2.3.4- tetrahydro-quinolin-6-yl}-4-tert-butyl-benzamide

According to general procedure C, thes compound described in example 31a (161 mg) was acylated with 4-fert-butylbenzoic acid (99 mg), DIPEA (322 pl) and HATU (210 mg) in CHzCl, (5 ml). The title compound was purified by preparative HPLC (method

A).

Yield: 80 mg (TFA salt); MS-ESI: [MH] = 598.2; HPLC: R; = 15.39 min (method 2).

Example 38

N-{1-Acetyl-2 2 4-trimethyl-4-[4-(2-mmorpholin-4-yl-ethoxy)-phenyl]-1,2.3.4- tetrahydro-quinolin-6-y1}-2.3-dichloro —benzamide

According to general procedure C, thes compound described in example 31a (161 mg) was acylated with 2,3-dichlorobenzoic acid (106 mg), DIPEA (322 pul) and HATU (210 mg) in CHCl, (5 ml). The title compound was purified by preparative HPLC (method

A). : Yield: 113 mg (TFA salt); MS-ESIL: [IM+H]" = 610.2; HPLC: R; = 11.42 min (method 2).

Example 39

N-{1-Acetyl-2,2.4-trimethyl-4-[4-(2-maorpholin-4-yl-ethoxy)-phenyl]-1.2.3.4- tetrahydro-quinolin-6-yl}-4-bromo-bemzamide

, According to general procedure C, the compound describecd in example 31a (260 mg) vvsas acylated with 4-bromobenzoic acid (179 mg), DIPEA_ (517 pl) and HATU (338 mg) in CH,Cl, (5 ml). The title compound was purified by preparative HPLC (method

A)

Wield: 127 mg (TFA salt); MS-ESI: [M+H]" = 620.2; HPL«C: R; = 12.24 min (method 2).

Example 40

MW-{1-Acetyl-2.2 4-trimethyl-4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-1.2,3.4- teetrahydro-quinolin-6-y1}-4-methoxy-3-methyl-benzamide

According to general procedure C, the compound described] in example 31a (260 mg) wvas acylated with 4-methoxy-3-methylbenzoic acid (148 mg), DIPEA (517 ul) and

HIATU (338 mg) in CH2Cl> (5 ml). The title compound vwas purified by preparative

HIPLC (method A).

Wield: 158 mg (TFA salt); MS-ESI: [M+H]" = 586.2; HPL«C: R, = 11.49 min (method 2)

Example 41 :

IV-{1-Acetyl-2.2 4-trimethyl-4-[4-(2-morpholin-4-yl-ethoxy )-phenyl]-1.2.3.4- teetrahydro-quinolin-6-yl}-4-dimethylamino-benzamide

According to general procedure C, the compound described in example 31a (260 mg) vevas acylated with 4-dimethylaminobenzoic acid (147 mg), BDIPEA (517 ul) and HATU (338 mg) in CHCl, (5 mi). The title compound was purified by preparative HPLC (Canethod A).

Wield: 95 mg (TFA salt); MS-ESI: [M+H]" = 585.2; HPLC: R,=9.53 min (method 2).

Example 42

INV-{1-Acetyl-2,2 4-trimethyl-4-[4-(2-morpholin-4-yl-ethoxy)-phenyl]-1.2.3.4- tetrahydro-quinolin-6-y1}-3-trifluoromethyl-benzamide ’ Wo a solution of the compound described in example 31a (C260 mg) and pyridine (500 1-1) in toluene (4.5 ml) was added 3-(trifluoromethyl)benzozyl chloride (185 mg). Ethyl acetate (15 ml) and water (15 ml) were added. The organic layer was separated and vwashed with water (15 ml), dried and concentrated in vacu=o. The title compound was purified by preparative HPLC (method A).

, Yield: 200 mg (TFA salt); MS-ESI: [M+H]" = 610.2; "HPLC: R, = 13.23 min (method

Example 43

N-{1-Acetyl-2 2 4-trimethyl-4-[4-(2-morpholin-4-yl-etihoxy)-phenyl}-1,2,3.4- tetrahydro-quinolin-6-yl}-3-nitro-benzamide

To a solution of the compound described in example 31a (260 mg) and pyridine (500 pb) in toluene (4.5 ml) was added 3-nitrobenzoyl chlor-ide (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 sak); MS-ESI: [M-+H]" = 587.4; lTHPLC: R, = 10.28 min (method 2).

Example 44

CHO-FSH in vitro bioactivity

FSH activity of compounds were tested in Chinese Hammster Ovary (CHO) cells stably transfected with the human FSH receptor and cotransfected with a cAMP responsive element (CRE) / promotor directing the expression of a firefly luciferase reporter gene.

Binding of ligand to the Gs-coupled FSH receptor will result in an increase of cAMP, which in turn will induce an increased transactivation of the luciferase reporter construct, To test antagonistic properties recombinant FSH in a concentration that induces approximately 80% of the maximal stimulatiom of cAMP accumulation in the absence of test compound was added (rec-hFSH; 10 mUJ/ml). The luciferase signal was quantified using a luminescence counter. For test compounds, ECso values (concentration of test compound causing half-maximal (50 %) stimulation or reduction) were calculated. For that purpose the software programm GraphPad PRISM, version 3.0 (GraphPad software Inc., San Diego) was used.

Compounds of all examples exhibited an ECs (ICsp) value of less than 10° M in either an agonistic or an antagonistic assay set-up or both. Thee compounds of examples 3, 4, - 30 7,10-13, 16, 36, 37, 39, 41 and 42 showed an ECs (ICs) of less than 107 M in at least one of the assays.

Claims (17)

Claims

1. A tetrahydroquinoline derivative according to Formula 1, oF @ ras N FL ke JN Formula I or a pharmaceutically acceptable salt thereof, wherein R' and R? are H, Me; R? is (2-6C)heterocycloalkyl( 1-4C)alkyl, (2-5C)beteroaryl(1 -4C)alkyl, (6C)aryl(1-4C)alkyl, (1-4C) cli)alkylaminocarbonylamino(2-4C)alkyl, (2- 6C)heterocycloalkylcarbonylamino(2-4C)alkyl, R*-(2-4C)alkyl or R>-carbonyl(1- 4C)alkyl; R* is (2-5C)heteroaryl, (6C)aryl, (3-8C)cycloatkyl, (2-6C)heterocycloalkyl or (1- 6C)alkyl RS is (di)(1-4C)alkylamino, (1-4C)alkoxy, amino, hydroxy, (6C)arylamino, (di)(3-4C)alkenylamino, (2-5C)heteroaryl(1-4C)alkylamino, (6C)aryl(1- 4C)alkylamino, {di){(1-4C)alkoxy(2-4C)alkyljamino, (di)[(1-4C)alkylamino(2- 4C)alkylJamino, (di)[amino(2-4C)alkyljamino or (di){hydroxy(2- 4C)alkyl]amino.

2. The derivative according to claim 1 wherein R? is (2-6C)heterocycloalkyl(1- 4C)alkyl, (2-5C)beteroaryl( 3-4C)alkyl, (2-6C)heterocycloalkylcarbonylamino(2- " 20 4C)alkyl, R*-(2-4C)alkyl or R*-carbonyl(1-4C)alkyl.

3. The derivative according to claims 1 or 2 wherein R? is (di)(1-4C)atkylamino, amino, (di)(3-4C)alkenylamino, (2-5C)beteroaryl(1-4C)alkylamino or (6C)aryl(1-4C)alkylamino

4. The derivative according to claims 1-3 wherein R’ is (di)(1-4C)alkylamino or amino.

PCT/EP2003/051024

5. The derivative according to claims {-4~ wherein R® is (di)(1-4C)alkylamino.

6. The derivative according to claims 1-5 wherein R* is (6C)aryl.

7. The derivative according to claims 1-6 wherein R® is (2-6C)heterocycloalkyl(1-4C)alkyl, (2--5C)heteroaryl(1-4C)alkyl or R3-(2- 4C)alkyl.

8. A pharmaceutical composition comprising the tetrahydroquinoline derivative of any one of claims 1-7 and pharmac ecutically suitable auxiliaries.

9. The tetrahydroquinoline derivative of” claims 1-7 for use in therapy.

10. Use of the tetrahydroquinoline deriva-tive of any one of claims 1-7 or a pharmaceutically acceptable salt or solvate thereof for the manufacture of a medicament for fertility regulation.

11. Use of the tetrahydroquinoline deriva. tive of claims 1-7 in the manufacture of a medicament for treating a disease, illEess, disorder or condition.

12. A substance or composition for use ir1 a method of fertility regulation, said substance or composition comprising the tetrahydroquinoline derivate of any one of claims 1-7 or a pharmaceutica_lly acceptable salt or solvate thereof, and said method comprising administerimg said substance or composition.

13. A derivative according to any one of claims 1 to 7, or 9, substantially as herein described and illustrated.

14. A composition according to claim 8, substantially as herein described and illustrated.

15. Use according to claim 10 or claim L 1, substantially as herein described and illustrated.

16. A substance or composition for use iin a method of treatment according to claim 12, substantially as herein described and illustrated.

17. A new derivative, a new composition, a new use of a derivative as claimed in any one of claims 1 to 7, or a substamce or composition for a new use in a method of treatment, substantially a s herein described. : AMENDED SHEET