HK1122285A - 2,4-diamino-pyrimidines used as aurora inhibitors - Google Patents

Description

2, 4-diamino-pyrimidines as Aurora inhibitors

The invention relates to novel 2, 4-diamino-pyrimidines of general formula (1)

Wherein the radical R¹To R³Having the meanings given in the claims and the description, and isomers thereof, processes for preparing these pyrimidines and their use as pharmaceutical compositions.

Background

Tumor cells evade regulation and control of the body, in whole or in part, and are characterized by uncontrolled growth. This is due on the one hand to the deletion of control proteins such as, for example, Rb, p16, p21 and p53 and on the other hand to the activation of cyclin-dependent kinases, so-called cell cycle promoters.

Studies in model organisms such as schizosaccharomyces pombe, drosophila melanogaster or xenopus laevis and in human cells have shown that the transition from the G2 phase to mitosis is regulated by CDK 1/cyclin B kinase (Nurse, 1990). This kinase, also known as "mitosis-promoting factor" (MPF), phosphorylates and regulates a variety of proteins, such as, for example, nuclear fiber layers, kinesin-like motor proteins, clusterin, and golgi matrix proteins, which play important roles in nuclear membrane disruption, centrosome separation, mitotic spindle conformation, chromosome condensation, and golgi disassembly (Nigg, 2001). Treatment of human tumor cells with inhibitors that inhibit CDK 1/cyclin B, such as, for example, butyrolactone, results in their arrest in the G2/M phase, followed by apoptosis (Nishio et al, 1996).

In addition to cyclin-dependent kinases, so-called polo-like serine/threonine kinases (PLK-1, PLK-2, PLK-3 and PLK-4) play an important role in regulating the cell cycle of eukaryotes. It has been found that PLK-1 in particular plays a nuclear role in regulating the mitotic phase. PLK-1 is responsible for the maturation of centrosomes, the activation of the phosphatase Cdc25C, and the activation of the anaphase promoting complex (Glover et al, 1998, Qian et al, 2001). Injection of PLK-1 antibody resulted in the arrest of non-transformed cells in the G2 phase, while tumor cells arrested in the mitotic phase (Lane and Nigg, 1996).

Furthermore, cessation at the G2/M phase may also be achieved by inhibition of specific motor proteins, so-called kinesins, such as, for example, Eg5(Mayer et al, 1999), or by agents which stabilize or destabilize tubulin (e.g., colchicine, taxol, etoposide, vinblastine, vincristine) (Schiff and Horwitz, 1980).

Serine/threonine kinases of the Aurora family regulate various processes of cell division. These processes include chromosome condensation, spindle dynamics, centromere-microtubule interactions, chromosome orientation, arrangement of metaphase plates, and cytokinesis (Meraldi et al, 2004; Carmena and Earnshaw, 2003; Andrews et al, 2003). Three members of this family, Aurora a, B and C, have been described in mammals. Aurora kinases of the a and B types are also present in c.elegans and drosophila melanogaster, whereas yeast contains only a single Aurora gene, designated IPL1 (in scorevisiae), or ARK1 (in s.pomber). All Aurora proteins share a similar overall structure, which includes a variable N-terminal, well-conserved central kinase domain and a short C-terminal portion. Despite their sequence similarity, Aurora family kinases also exhibit different subcellular localization associated with specific functions.

Therefore, Aurora a will be found in interphase centrosomes as well as on mitotic centrosomes and spindle tubulin near poles. Thus-as confirmed by RNA interference experiments-Aurora a is essential for entry into mitosis, as centrosome maturation and segregation cannot occur when Aurora a is absent. A wide variety of Aurora A activators are available, such as, for example, TPX2, Ajuba or protein phosphatase inhibitor-2. TPX2 appears to be responsible for the spatio-temporal correct activation of Aurora A on spindle tubulin close to polar (Hirota et al, 2003; Bayliss et al, 2003; Eyersan Maller, 2004; Kufer et al, 2002; Satinover et al, 2004).

Aurora B is associated with condensed chromosomes in the early prophase, localizes to the centromere in the metaphase, thereafter relocates to the central region of the central spindle, and finally condenses on the so-called fleming or centrsome at cytokinesis, i.e. the narrowly defined region between daughter cells. These characteristic spatial changes in mitosis demonstrate that Aurora B is a so-called "chromosomal messenger" protein. At least three other "chromosomal messenger" proteins are known to form complexes with Aurora B. They are INCENP (internal centromeric protein), survivin and borealin (Andrews et al, 2003; Carmena and Earnshaw, 2003; Meraldi et al, 2004). The C-terminus of INCENP provides an important point of contact between Aurora B and the complex, the so-called "IN-box". "IN-box" is the most conserved region of INCENP. It binds to and activates Aurora B and is phosphorylated by this kinase (Adams et al, 2000; Bishop and Schumacher, 2002; Kaitna et al, 2000; Bolton et al, 2002; Honda et al, 2003).

Aurora C is the least characteristic member of the Aurora family. Aurora C also binds to INCENP and behaves as a "chromosomal messenger" protein, although it has the highest expression level after Aurora B. It is speculated that Aurora C may be able to replace some of the functions of Aurora B, such as, for example, that expression of Aurora C could normalize multinuclear phenotype Aurora B-depleted cells (Sasai et al, 2004; Li et al, 2004).

Aurora B phosphorylates histone H3 at Ser10 and Ser 28. Although this phosphorylation is consistent with chromosome condensation time, the effects of this event are only associated with later stages of the cell cycle. This is confirmed by the fact that histone H3 concentrates in the mitotic chromosome with phosphorylation of Ser10 on heterochromatin near the centromere and concomitant triple methylation of Lys 9. This modified histone H3 prevents the binding of heterochromatin protein 1(HP1) by a "chromosomal messenger" protein complex, allowing access to the centromere region of the centromere (Hirota t et al, Manuscript in preparation).

One function of Aurora B is that it appears by inhibiting Aurora B, combining different proteins at the centromere in the metaphase (Ditchfield et al, 2003; Hauf et al, 2003; Murata-Hori and Wang, 2002; Vigneron et al, 2004). Aurora B plays a central role in the signaling pathway for detecting and correcting centromeric (defective because they start only at one spindle pole) centromere appendages of microtubules (Andrews et al, 2003; Carmena and Earnshaw, 2003; Meraldi et al, 2004). If the attachment state is not corrected, errors will occur in chromosome segregation. Aurora B-mediated phosphorylation of microtubule depolymerase MCAK is associated with this correction mechanism (Gorbsky, 2004).

Aurora B also phosphorylates proteins important for the formation of replicative and cytokinesis, such as, for example, mgcrac gap, the regulatory light chain of myosin II, vimentin, desmin, GFAP (glial fibrillary acidic protein), and the driver proteins MKLP1 and MKLP2, where MKLP2 is presumed to be responsible for completing the transfer of the "chromosomal messenger" protein complex from the centromere to the centrosome (Gruneberg et al, 2004).

In view of the various functions of Aurora B in the cell cycle, it was surprisingly found that inhibiting Aurora B in tumor cells does not cause mitotic arrest, but rather allows the cell cycle to continue without cytokinesis (Hauf et al, 2003). Due to the accumulation of homocentromeric microtubule-centromere appendages and the resulting erroneous chromosomal segregation, a large number of polyploid polyploidia appear, eventually leading to apoptosis. Even simultaneous inhibition of Aurora a did not affect this phenotype (Keen and Taylor, 2004).

Initially, it was mainly Aurora A that showed oncogenic activity (e.g., transformation of murine fibroblasts after overexpression), however, for Aurora B, these signs were only indirect (Zhou et al, 1998; Bischoff et al, 1998; Katayama et al, 1999). The recognition that overexpression of Aurora B in embryonic hamster cells and its use in xenograft experiments directly increases the incidence, size and invasiveness of tumors has changed. The corresponding tumors show chromosomal instability and increased histone H3 Ser10 phosphorylation (Ota et al, 2002). These results support the importance of AuroraB in tumorigenesis.

Pyrimidines are generally known as inhibitors of kinases. Thus, for example, substituted pyrimidines with a nonaromatic group in the 4-position are described in international patent applications WO 02/096888 and WO 03/032997 as active ingredients with anticancer action.

The object of the present invention is to indicate novel active substances which can be used for the prophylaxis and/or treatment of diseases which are characterized by excessive or abnormal cell proliferation.

Detailed Description

Surprisingly, it has now been found that compounds of the general formula (1) in which the radical R¹、R²And R³As defined later, act as inhibitors of specific cell cycle kinases. Thus, the compounds of the invention are useful, for example, in the treatment of diseases associated with specific cell cycle kinase activity and characterized by excessive or abnormal cell proliferation.

The invention relates to compounds of general formula (1)

Wherein

R¹Is represented by R⁵And optionally substituted by one or more R⁴A substituted group selected from C_3-10-cycloalkyl and 3-8 membered heterocycloalkyl;

R²represents optionally substituted by one or more R⁴A substituted group selected from C_1-6Alkyl radical, C_3-10-cycloalkyl, 3-8 membered heterocycloalkyl, C_6-15-aryl and 5-12 membered heteroaryl;

R³represents hydrogen, halogen, -CN, -NO₂、C_1-4Alkyl radical, C_1-4-haloalkyl group, C_3-10-cycloalkyl, C_4-16-cycloalkylalkyl and C_7-16-a radical of arylalkyl;

R⁴represents a group selected from R^a、R^bAnd by one or more R, which may be the same or different^cAnd/or R^bSubstituted R^aA group of (a);

R⁵represents a group selected from-C (O) R^c、-C(O)NR^cR^c、-S(O)₂R^c、-N(R^f)S(O)₂R^c、-N(R^f)C(O)R^c、-N(R^f)C(O)OR^cand-N (R)^f)C(O)NR^cR^cA group of (a);

each R^aIndependently of one another, from C_1-6Alkyl radical, C_3-10-cycloalkyl, C_4-16-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl;

each R^bAre suitable radicals, in each case independently of one another, selected from ═ O, -OR^c、C_1-3Haloalkoxy, -OCF₃、＝S、-SR^c、＝NR^c、＝NOR^c、-NR^cR^cHalogen, -CF₃、-CN、-NC、-OCN、-SCN、-NO₂、-S(O)R^c、-S(O)₂R^c、-S(O)₂OR^c、-S(O)NR^cR^c、-S(O)₂NR^cR^c、-OS(O)R^c、-OS(O)₂R^c、-OS(O)₂OR^c、-OS(O)₂NR^cR^c、-C(O)R^c、-C(O)OR^c、-C(O)NR^cR^c、-CN(R^f)NR^cR^c、-CN(OH)R^c、-CN(OH)NR^cR^c、-OC(O)R^c、-OC(O)OR^c、-OC(O)NR^cR^c、-OCN(R^f)NR^cR^c、-N(R^f)C(O)R^c、-N(R^f)C(S)R^c、-N(R^f)S(O)₂R^c、-N(R^f)C(O)OR^c、-N(R^f)C(O)NR^cR^c、-[N(R^f)C(O)]₂R^c、-N[C(O)]₂R^c、-N[C(O)]₂OR^c、-[N(R^f)C(O)]₂OR^cAnd N (R)^f)CN(R^f)NR^cR^c；

Each R^cIndependently of one another, hydrogen or optionally substituted by one or more identical or different R^dAnd/or R^eSubstituted groups selected from: c_1-6Alkyl radical, C_3-10-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl,

each R^dIndependently of one another, hydrogen or optionally substituted by one or more identical or different R^eAnd/or R^fSubstituted groups selected from: c_1-6Alkyl radical, C_3-8-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl;

each R^eAre suitable radicals and are each, independently of one another, selected from ═ O, -OR^f、C_1-3Haloalkoxy, -OCF₃、＝S、-SR^f、＝NR^f、＝NOR^f、-NR^fR^fHalogen, -CF₃、-CN、-NC、-OCN、-SCN、-NO₂、-S(O)R^f、-S(O)₂R^f、-S(O)₂OR^f、-S(O)NR^fR^f、-S(O)₂NR^fR^f、-OS(O)R^f、-OS(O)₂R^f、-OS(O)₂OR^f、-OS(O)₂NR^fR^f、-C(O)R^f、-C(O)OR^f、-C(O)NR^fR^f、-CN(R^g)NR^fR^f、-CN(OH)R^f、-C(NOH)NR^fR^f、-OC(O)R^f、-OC(O)OR^f、-OC(O)NR^fR^f、-OCN(R^g)NR^fR^f、-N(R^g)C(O)R^f、-N(R^g)C(S)R^f、-N(R^g)S(O)₂R^f、-N(R^d)C(O)OR^f、-N(R^g)C(O)NR^fR^fAnd N (R)^g)CN(R^f)NR^fR^f；

Each R^fIndependently of one another, hydrogen or optionally substituted by one or more identical or different R^gSubstituted groups selected from: c_1-6Alkyl radical, C_3-8-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl;

each R^gIndependently of one another are hydrogen, C_1-6Alkyl radical, C_3-8-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl, optionally in the form of tautomers, racemates, enantiomers, diastereomers and mixtures thereof, and optionally in the form of pharmaceutically acceptable acid addition salts thereof.

One aspect of the present invention relates to compounds of formula (1) wherein R³Represents a group selected from halogen and C_1-4A haloalkyl group.

Another aspect of the invention relates to compounds of formula (1), wherein R³represents-CF₃。

Another aspect of the invention relates to compounds of formula (1), wherein R²Represents optionally substituted by one or more R⁴Substituted C_6-10Aryl or 5-12 membered heteroaryl.

Another aspect of the present invention relates to formula (1)) Wherein R is²Represents optionally substituted by one or more R⁴A substituted phenyl group.

Another aspect of the invention relates to compounds of the general formula (1A),

wherein

n is equal to 0 or 1, and

m is equal to 1-5, and

y is equal to 0 to 6, and the remaining groups are as defined above.

Another aspect of the invention relates to compounds of formula (1A), wherein R³Represents a group selected from halogen and C_1-4A haloalkyl group.

Another aspect of the invention relates to compounds of formula (1A), wherein R³Denotes CF₃。

Another aspect of the invention relates to compounds of formula (1A), wherein R²Represents optionally substituted by one or more R⁴Substituted C_6-10Aryl or 5-12 membered heteroaryl.

Another aspect of the invention relates to compounds of formula (1A), wherein R²Represents optionally substituted by one or more R⁴A substituted phenyl group.

Another aspect of the present invention relates to a compound of formula (1) or (1A), or a pharmaceutically active salt thereof, for use as a pharmaceutical composition.

Another aspect of the present invention relates to a compound of formula (1) or (1A), or a pharmaceutically active salt thereof, for use in the preparation of a pharmaceutical composition having antiproliferative activity.

Another aspect of the present invention relates to pharmaceutical preparations which comprise, as active substance, one or more compounds of the general formula (1) or (1A) or physiologically acceptable salts thereof, optionally in association with customary excipients and/or carriers.

Another aspect of the present invention relates to the use of a compound of formula (1) or (1A) for the preparation of a pharmaceutical composition for the treatment and/or prevention of cancer, infection, inflammation and autoimmune diseases.

Another aspect of the present invention relates to pharmaceutical preparations comprising a compound of general formula (1) or (1A) and at least one other cytostatic or cytotoxic active substance different from general formula (1), optionally in the form of tautomers, racemic compounds, enantiomers, diastereomers and mixtures thereof, and optionally the pharmacologically acceptable acid addition salts thereof.

Definition of

The following definitions apply herein unless otherwise indicated.

Alkyl substituents refer in each case to saturated, unsaturated, straight-chain or branched aliphatic hydrocarbon radicals (alkyl radicals), and this definition includes saturated alkyl radicals and unsaturated alkenyl radicals as well as alkynyl radicals. The alkenyl substituents are in each case straight-chain or branched, unsaturated alkyl groups having at least one double bond. Alkynyl substituents in each case denote straight-chain or branched, unsaturated alkyl radicals having at least one triple bond.

Heteroalkyl denotes a straight or branched aliphatic hydrocarbon chain comprising 1 to 3 heteroatoms, and the individual carbons and heteroatoms available in the heteroalkyl chain may each optionally be substituted independently of one another, and the heteroatoms are selected independently of one another from O, N, P, PO₂S, SO and SO₂(e.g., dimethylaminomethyl, dimethylaminoethyl, dimethylaminopropyl, diethylaminomethyl, diethylaminoethyl, diethylaminopropyl, 2-diisopropylaminoethyl, bis-2-methoxyethylamino, [2- (dimethylamino-ethyl) -ethyl-amino]-methyl, 3- [2- (dimethylamino-ethyl) -ethyl-amino]-propyl, hydroxymethyl, 2-hydroxyethyl, 3-hydroxypropyl, methoxy, ethoxy, propoxy, methoxymethyl, 2-methoxyethyl).

Haloalkyl refers to an alkyl group in which one or more hydrogen atoms are replaced by halogen atoms. Haloalkyl includes saturated alkyl and unsaturated alkenyl and alkynyl groups, such as, for example, -CF₃、-CHF₂、-CH₂F、-CF₂CF₃、-CHFCF₃、-CH₂CF₃、-CF₂CH₃、-CHFCH₃、-CF₂CF₂CF₃、-CF₂CH₂CH₃、-CF＝CF₂、-CCl＝CH₂、-CBr＝CH₂、-CJ＝CH₂、-C≡C-CF₃、-CHFCH₂CH₃and-CHFCH₂CF₃。

Halogen means a fluorine, chlorine, bromine and/or iodine atom.

Cycloalkyl refers to mono-or polycyclic rings wherein the ring system may be saturated or unsaturated, non-aromatic or spiro compounds, optionally containing double bonds, such as, for example, cyclopropyl, cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, cycloheptenyl, norbornyl, norbornenyl, indanyl, adamantyl, spiroheptanyl and spiro [4.2] heptanyl.

Cycloalkylalkyl groups include acyclic alkyl groups in which a hydrogen atom bonded to a carbon atom is replaced with a cycloalkyl group.

Aryl means monocyclic or bicyclic rings having 6 to 12 carbon atoms, such as, for example, phenyl and naphthyl.

Arylalkyl includes acyclic alkyl groups in which a hydrogen atom bonded to a carbon atom is replaced with an aryl group.

Heteroaryl refers to a mono-or polycyclic ring containing one or more heteroatoms in place of one or more carbon atoms, which are the same or different, and are, for example, nitrogen, sulfur or oxygen atoms. Examples include furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, and triazinyl. Examples of bicyclic heteroaryls are indolyl, isoindolyl, benzofuranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl, indazolyl, isoquinolyl, quinolinyl, quinoxalinyl, cinnolinyl, 2, 3-naphthyridinyl, quinazolinyl and benzotriazinyl, indolizinyl, oxazolopyridinyl, imidazopyridinyl, 1, 5-naphthyridinyl, indolinyl, isobenzohydropyranyl, chromanyl, tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuryl, isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl, pyridopyridyl, benzotetrahydrofuranyl, benzothiophenyl, purinyl, benzodioxolyl, triazinyl, benzoxazinyl, phenothiazinyl, benzothiazolyl, benzoxazolyl, benzisothiazolyl, 1, 5-diazepinyl, dihydropyranyl, 1, 5-diazepinyl, 1-indolyl, and the like, Pteridinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl, dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl, dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl, coumarinyl, isocoumarinyl, chromonyl, chromanone, pyridyl-N-oxide, tetrahydroquinolyl, dihydroquinolyl, dihydroquinolinone, dihydroisoquinolinonyl, dihydrocoumarinyl, dihydroisocoumarinyl, isoindolinonyl, benzodioxan, benzoxazolinonyl, pyrrolyl-N-oxide, pyrimidinyl-N-oxide, pyridazinyl-N-oxide, pyrazinyl-N-oxide, quinolyl-N-oxide, indolyl-N-oxide, indolinyl-N-oxide, isoquinolinyl-N-oxide, quinazolinyl-N-oxide, quinoxalinyl-N-oxide, 2, 3-naphthyridinyl-N-oxide, imidazolyl-N-oxide, isoxazolyl-N-oxide, oxazolyl-N-oxide, thiazolyl-N-oxide, indolizinyl-N-oxide, indazolyl-N-oxide, benzothiazolyl-N-oxide, benzimidazolyl-N-oxide, pyrrolyl-N-oxide, oxadiazolyl-N-oxide, thiadiazolyl-N-oxide, triazolyl-N-oxide, tetrazolyl-N-oxide, thiadiazolyl-N, benzothiopyranyl-S-oxide and benzothiopyranyl-S, S-dioxide.

Heteroarylalkyl groups include acyclic alkyl groups in which a hydrogen atom bonded to a carbon atom is replaced with a heteroaryl group.

Heterocyclyl relates to saturated or unsaturated, non-aromatic, monocyclic, bicyclic or bridged polycyclic or spiro compounds comprising 3 to 12 carbon atoms and also carrying heteroatoms, such as nitrogen, oxygen or sulphur, in place of one or more carbon atoms. Examples of such heterocyclic groups are tetrahydrofuranyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, pyrazolidinyl, pyrazolinyl, piperidinyl, piperazinyl, indolinyl, isoindolinyl, morpholinyl, thiomorpholinyl, homomorpholinyl, homopiperidinyl, homopiperazinyl, homothiomorpholinyl, thiomorpholinyl-S-oxide, thiomorpholinyl-S, S-dioxide, tetrahydropyranyl, tetrahydrothienyl, homothiomorpholinyl-S, S-dioxide, oxazolidonoyl, dihydropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidyl, dihydrofuranyl, dihydropyranyl, tetrahydrothienyl-S-oxide, tetrahydrothienyl-S, S-dioxide, homothiomorpholinyl 1-S-oxide, pyrazolidinyl, S-oxide, thiomorpholinyl-S-oxide, dihydropyranyl-S-oxide, 2-oxa-5-azabicyclo [2.2.1] heptane, 8-oxa-3-aza-bicyclo [3.2.1] octane, 3, 8-diaza-bicyclo [3.2.1] octane, 2, 5-diaza-bicyclo [2.2.1] heptane, 3, 8-diaza-bicyclo [3.2.1] octane, 3, 9-diaza-bicyclo [4.2.1] nonane and 2, 6-diaza-bicyclo [3.2.2] nonane.

Heterocycloalkylalkyl refers to acyclic alkyl groups in which a hydrogen atom bonded to a carbon atom is replaced with a heterocycloalkyl group.

Abbreviation list

Eq.eq	Equivalent weight	IR	Infrared spectroscopy
				Ac	Acetyl group	Cat.cat	Catalysts, catalyzed
Boc	Tert-butyloxycarbonyl radical	conc.	Concentrated
				Bu	Butyl radical	B.p.b.p.	Boiling point
BuLi	N-butyl lithium	LC	Liquid chromatography
				c	Concentration of	Hünig base	N-ethyl-diisopropylamine
cHex	Cyclohexane	i	Different from each other
				CDI	Carbonyl diimidazoles	mCPBA	Meta-chloroperoxybenzoic acid
CSI	Chlorosulfonyl isocyanates	min	Minute (min)
				DC，TLC	Thin layer chromatography	Me	Methyl radical
DCC	Dicyclohexylcarbodiimide	MS	Mass spectrometry
				DCM	Methylene dichloride	NMP	N-methyl pyrrolidone
DIPEA	Ethyl diisopropylamine (Hunig alkali)	NMR	Nuclear magnetic resonance
				DMAP	N, N-dimethylaminopyridine	Ph	Phenyl radical
DMF	N, N-dimethylformamide	Pr	Propyl radical
				DMA	N, N-dimethyl acetamide	rac	Racemic
DMSO	Dimethyl sulfoxide	Rf(Rf)	Retention factor
				EE	Ethyl acetate	RP	Inverse phase
ESI	Electrospray ionization	RT	Room temperature or Retention time (HPLC)
				Et	Ethyl radical	t	Tertiary amine
h	Hour(s)	THF	Tetrahydrofuran (THF)
				hex	Hexyl radical	TBTU	O- (benzotriazol-1-yl) -N, N, N ', N' -tetramethyl-uronium tetrafluoroborate
HPLC	High performance liquid chromatography	UV	Ultraviolet light

LDA

Lithium diisopropylamide

The following examples are intended to illustrate the invention without limiting its scope.

General description of the invention

All reactions were carried out in commercial equipment by conventional methods used in chemical laboratories unless otherwise indicated.

The solvent used was purchased as analytical grade and used without further purification. All reagents were used directly without purification during the synthesis.

The starting materials sensitive to air and/or humidity are stored under argon and the corresponding reactions and operations using them are carried out under a protective gas (nitrogen or argon).

Chromatography

For the preparation of media pressure chromatography (MPLC, normal phase), Silica gel manufactured by Millipore (name: Granula Silica Si-60A 35-70 μm) or C-18 RP-Silica gel (RP-phase) manufactured by Macherey Nagel (name: Polygonrep 100-50C 18) was used.

Thin layer chromatography was performed on slides produced by Merck using silica gel 60 TLC plates (with fluorescent indicator F-254).

For preparative high pressure chromatography (HPLC) analytical HPLC (reaction control) was performed using a column manufactured by Waters (name: XTerraPrep. MS C18, 5 μ M, 30. mu.100 mm or XTerra Prep. MS C18, 5 μ M, 50. mu.100 mmOBD or Symmetry C18, 5 μ M, 19. mu.100 mm) using a column manufactured by Agilent (name: Zorbax SB-C8, 5 μ M, 21.2. mu.50 mm).

For chiral high pressure chromatography (HPLC), a column (name: Chiralpak AD-H or Chiralpak AS or Chiracel OD-RH or Chiracel OD-H or Chiracel OJ-H, various sizes and 5 μm material) manufactured by Daicel Chemical Industries, Ltd.

Nuclear Magnetic Resonance (NMR)

Nuclear magnetic resonance was performed in deuterated dimethyl sulfoxide-d 6 as a solvent. If other solvents are used, they are explicitly mentioned in the examples or methods. Chemical shifts are referenced to standard tetramethylsilane (δ ═ 0.00 ppm). The measurement was obtained using Avance400(400MHz-NMR-spectrometer) or Avance 500(500 MHz-NMR-spectrometer) manufactured by Bruker Biospin GmbH.

HPLC-Mass Spectrometry/UV-Spectroscopy

HPLC-MS apparatus (high Performance liquid with Mass Detector) made by Agilent produces characteristic example retention time/MS-ESI⁺。

The instrument was assembled with a diode array detector (G1315B manufactured by Agilent) and a mass detector (1100 LS-MSD SL; G1946D; Agilent) connected in series downstream of the chromatograph (column: XTerra MSC18, 2.5 μm, 2.1 × 30mm, Waters or Synergi POLAR-RP 80A; 4 μm, Phenomenex).

The instrument was run at a flow rate of 1.1 ml/min. For the separation process, a gradient was run over 3.1 minutes (start of gradient: 95% water and 5% acetonitrile; end of gradient: 5% water and 95% acetonitrile; in each case 0.1% formic acid was added to both solvents).

Melting Point

Melting points were obtained using a B-540 apparatus manufactured by B ü chi and were not corrected.

When the preparation of the starting compounds is not described, they are either commercially available or can be similarly made into known compounds or prepared by the methods described herein.

Preparation of the Compounds of the invention

The compounds of the present invention having substituents of the above general formula can be prepared by the following synthetic methods. These methods are intended to describe the invention, not to limit the illustrative objects and not to limit the scope of the claimed compounds to these examples.

Procedure A

Procedure B

Optionally, after formation of the diaminopyrimidine, conversion of one or more functional groups is also possible.

Procedure C

Optionally, after formation of the diaminopyrimidine, conversion of one or more Functional Groups (FG) is also possible. This is described in the related embodiment.

Procedure D

Procedure E

Preparation of the starting Compounds

Unless otherwise stated, all starting materials were purchased from suppliers and used directly in the synthesis. The substances described in the literature are prepared according to published synthetic methods.

A-1)2, 4-dichloro-5-trifluoromethyl-pyrimidine

48g (267mmol) of 5-trifluoromethyluracil are suspended in 210mL of phosphorus oxychloride (POCl)₃) And simultaneously, moisture is isolated. 47.7g (320mmol, 1.2eq) diethylaniline were slowly added dropwise to the suspension, the temperature being kept between 25 ℃ and 30 ℃. After the addition is complete, the mixture is stirred in a water bath for 5-10 minutes and then heated at 80-90C for 5-6 hours while excluding moisture. Excess POCl was removed by stirring in ice water containing about 1200g sulfuric acid₃Then in each caseThe aqueous phase is then extracted 3 times immediately, using 500ml of diethyl ether or tert-butyl-methyl-ether. The combined ether extracts were washed 2 times with ice water containing 300mL sulfuric acid (ca. 0.1M) and cold brine solution and then immediately dried over sodium sulfate. The drying agent was filtered off and the solvent was removed in vacuo. The residue is distilled in vacuo (10mbar) through a short column (20cm) (head temperature: 65-70 ℃) to give 35.3g (0.163mol, 61%) of a colorless liquid which is poured off and stored under argon.

DC：R_f＝0.83(cHex∶EE＝3∶1)

A-2) 2-chloro-4-methylsulfanyl-5-trifluoromethyl-pyrimidine and

a-3) 4-chloro-2-methylsulfanyl-5-trifluoromethyl-pyrimidine

5g (23mmol) of 2, 4-dichloro-5-trifluoromethyl-pyrimidine are dissolved in 40mL of THF, the solution is brought to-25 ℃ and 1.8g (25.3mmol, 1.1eq) of sodium thiomethoxide are added. The mixture was stirred at-25 ℃ for 1 hour and then at room temperature overnight without cooling. It was then diluted with dichloromethane and washed 3 times with 1N HCl. The organic phase is dried over magnesium sulfate and then evaporated to dryness in vacuo. The crude product was purified by column chromatography (silica gel, cyclohexane/dichloromethane; from 90/10 to 80/20% in about 20 minutes). 1.56g (6.8mmol, 30%) of product A-3 and 1.46g (6.4mmol, 28%) of product A-2 are isolated as colorless oil. Furthermore, 0.24g (4%) of 2, 4-bis-methylsulfanyl-5-trifluoromethyl-pyrimidine are isolated as a colorless solid.

Product A-3 product A-2

R_f(cHex∶CH₂Cl₂ 1∶1) 0.48 0.40

Structural analysis was done by chemical derivatization followed by NMR spectroscopy. For this, A-2 and A-3 were first brought at 100 ℃ in THF at 5bar H₂Pd/C and Pd (OH) in a ratio of 1: 1₂In each case dehalogenated separately. Due to the different symmetry of the products formed, it is possible to clearly identify regioisomers.

4-amino-N-methyl-N-phenyl-benzenesulfonamide (educt from example 1)

9.5mL (85.7mmol, 98%) of N-methylaniline are dissolved in 100mL of dichloromethane, 20g (85.7mmol, 95%) of 4-nitrobenzenesulfonyl chloride dissolved in 150mL of dichloromethane are added dropwise at 0 ℃ and the mixture is stirred for 1.5 hours. The organic phase was washed with saturated aqueous sodium carbonate solution and dried over sodium sulfate. Finally, it is filtered through silica gel and, after removing all volatiles in vacuo, 24.6g of crude N-methyl-4-nitro-N-phenyl-benzenesulfonamide are obtained.

14.6g (49.9mmol) of the nitrosulfonamide are dissolved in 100mL of THF/MeOH 1/1. After addition of Pd/C (10%), the mixture was brought to 5bar H at 50C₂Stirred under pressure for 16 hours. Adding molecular sieve to bind water, further adding Pd/C, and hydrogenating under hydrogenation conditions (5bar H)₂Stirring under pressure at 60 ℃ for a further 16 h gave 13.1g (48.9mmol, 100%) of crude A-4a as a pale brown solid. This crude product was used for the synthesis without any further purification.

4-amino-N-phenyl-benzenesulfonamide and 4-amino-N, N-dimethyl-benzenesulfonamide (analogs in examples 2 and 3) were prepared analogously. The process is generally applicable to the preparation of substituted or unsubstituted aminobenzenesulfonic acid amides from the corresponding nitrobenzenesulfonic acid chlorides.

General procedure for the Synthesis of Compounds of type B-2

The corresponding R3-substituted 2, 4-dichloropyrimidine B-1 (commercial or prepared by chlorinating the corresponding uracil as described in the a-1 example) was dissolved in THF (or dioxane, DMA, NMP, acetone) (about 2-5mL pro mmol), 1-1.6eq hunig base (or triethylamine, potassium carbonate or another suitable base) was added and the temperature of the reaction mixture was adjusted (at-78 ℃ for very active pyrimidines, RT or elevated temperature for slightly inactive pyrimidines). About 0.75 to 1eq of amine dissolved in the corresponding solvent (see above) are then added and the reaction mixture is stirred at the corresponding temperature for a specific time or melted or heated for a specific time, depending on the reactivity of the pyrimidine used. After the reaction was complete (reaction monitored by HPLC or DC), the reaction mixture was combined with silica gel and all volatiles were removed in vacuo. Purifying by column chromatography to obtain the desired substituted product. Depending on the radical R3 of the pyrimidine, two possible regioisomers are obtained in different ratios. They can generally be separated chromatographically.

B-2a)(±)-(1S^*，2R^*) -2- (2-chloro-5-trifluoromethyl-pyrimidin-4-ylamino) -cyclopentanecarboxamide

500mg (2.3mmol) of A-1 and 636mg (4.6mmol, 2eq) of potassium carbonate are suspended in 11mL of acetone, cooled to-70 ℃ and then cis- (+ -) - (1S, 2R) -2-amino-cyclopentanecarboxamide is added. The reaction was allowed to freeze-thaw to room temperature and stirred overnight, then stirred at room temperature for an additional 24 hours. Then 40mL of silica gel was added and all volatiles were removed in vacuo. The two regioisomeric products were separated by column chromatography, the desired regioisomer being the product which was first eluted (silica gel, cHex/EE 40/60). 218mg (0.71mmol, 31%) of B-2a and 297mg (0.96mmol, 42%) of the regioisomeric product B-2' a are separated.

R_f(B-2a) ═ 0.51 (silica gel, EE), [ R-_f(B-2a’)＝0.34]

MS-ESI+：309(M+H)⁺

The structures of these two regioisomers were identified and classified by dehalogenation of the product under reducing conditions and subsequent 1H-NMR-spectroscopy, respectively (analogously to A-2 and A-3).

The following examples of compounds of type B-2 were synthesized analogously.

#	R3	Condition	B-2∶B-2’	Yield B-2	Rf(B-2)	Rf(B-2’)	Eluent
								B-2a	CF3	Acetone, K2CO3，-70℃-RT16 hours, the time is	42∶58	31％	0.51	0.34	EE
B-2b	Me	DMA, Hunig's base, 40 ℃, 24 hours	＞85∶15	83％	0.25	Not detected	EE
								B-2c	NO2	Acetone, K2CO316 hours at-70 ℃ C	＞99∶1	82％	0.54	---	EE

B-2d	F	Dichloromethane, Hunig's base, 0 deg.C-RT, 2 days	＞99∶1	82％	0.43	---	EE
								B-2e	Cl	Dichloromethane, Hunig's base, 0 deg.C-RT, 1 day	Not detected	60％	0.45	Not detected	EE
B-2f	i-Pr	DMA, Hunig base, 70 ℃, 24 hours	Not detected	60％	0.40	0.28	EE

The compounds of B-2a to B-2f can be reacted with aniline under acid catalysis to form compounds of type B-4.

General procedure for the Synthesis of Compounds of type B-4

The educt B-2 is dissolved in 1-butanol (or dioxane, DMA, NMP) (about 0.5-4mL/mmol), 0.1-1eq of HCl in dioxane is added and 1eq of aniline and the reaction mixture are refluxed. After the reaction was complete, the reaction mixture was combined with silica gel and all volatiles were removed in vacuo. The mixture was then purified by column chromatography. Generally, when the reaction is completed, the product is precipitated from the reaction solution, which may be directly filtered with suction and washed with 1-butanol.

#	R3	Condition	Yield B-4	Rf	Eluent
						B-4a	CF3	Preparation from C-1 according to scheme C (C-3a ≡ B-4a)	--	0.37	DCM∶MeOH∶AcOH9∶1∶0.1
B-4b	Me	1-Butanol, 0.1eq HCl, refluxing for 3 hours	95％	0.11	DCM∶MeOH∶AcOH9∶1∶0.1

B-4c	NO2	1-Butanol, 0.1eq HCl, refluxing for 4 hours	66％	Not detected	---
						B-4d	F	1-Butanol, 0.1eq HCl, refluxing for 4 hours	83％	0.27	DCM∶MeOH∶AcOH9∶1∶0.1
B-4e	Cl	1-Butanol, 0.1eq HCl, refluxing2 hours	92％	0.31	DCM∶MeOH∶AcOH9∶1∶0.1
						B-4f	i-Pr	1-Butanol, 0.1eq HCl, refluxing for 4 hours	99％	0.08	DCM∶MeOH∶AcOH9∶1∶0.1

(4-amino-2-chloro-phenyl) - (4-methyl-piperazin-1-yl) -methanone (educt from example 70)

1mL (8.84mmol, 1.3eq) of N-methylpiperazine was dissolved in 40mL of dichloromethane, and this solution was combined with 1.5mL (8.84mmol, 1.3eq) of Hunig base. Then 1-5g (6.82mol, 1eq) of 4-nitro-2-chlorobenzoyl chloride dissolved in 10mL of dichloromethane are slowly added dropwise with cooling. After 2 hours, 9mL of saturated aqueous sodium bicarbonate solution were slowly added dropwise with stirring, the organic phase was separated and the solvent was removed in vacuo. The product is purified by column chromatography (silica gel, DCM/MeOH/NH)₃9/1/01) to yield 1.83g (6.45mmol, 95%) of nitrobenzamide. The latter was dissolved in 21 THF, 300mg of Raney nickel were added and the mixture was stirred at 3bar H₂Stirred under pressure at room temperature for 16 hours. After filtering off the raney nickel, the volatiles are removed in vacuo to yield 1.2g (4.73mmol, 73%) (4-amino-2-chloro-phenyl) - (4-methyl-piperazin-1-yl) -methanone.

R_f0.38 (silica gel, DCM: MeOH: NH)₃＝9∶1∶0.1)

MS-ESI⁺：254(M+H)⁺

Similarly, the method is suitable for the synthesis of substituted and unsubstituted aminobenzamides such as used in the syntheses of examples 71-75. These examples were prepared analogously to example 70. In the syntheses of examples 106, 107 and 144, m-aminobenzamides prepared in the same way are used.

Cis- (+/-) -2-amino-cyclopentanecarboxylic acid isopropylamide

55mg (0.43mmol) of cis- (. + -.) -2-amino-cyclopentanecarboxylic acid were suspended in 900. mu.L (25eq) of isopropylamine and 205mg (0.064mmol, 1.5eq) of TBTU and 550. mu.L of DMF were added to this suspension. It was stirred for 16 hours and the reaction mixture was taken up in DCM: MeOH: NH₃9: 1: 0.1, combined with 7mL of silica gel. After removal of all volatiles in vacuo, the mixture was chromatographed (silica gel DCM: MeOH: NH)₃9: 1: 0.1). 63mg (0.37mmol, 86%) of a colorless solid are obtained.

R_f0.33 (silica gel, DCM: MeOH: NH)₃ 85∶15∶1.5)

B-2g)(±)-(1S^*，2R^*) -2- (2-chloro-5-trifluoromethyl-pyrimidin-4-ylamino) -cyclopentanecarboxylic acid isopropylamide

2g (9.2mmol) of A-1 and 1.8mL (11.2mmol, 1.2eq) of Hunig's base are dissolved in 60mL of THF, the mixture is cooled to-78 ℃ and cis- (. + -.) -2-amino-cyclopentanecarboxylic acid isopropylamide dissolved in 60mL of THF is then added slowly dropwise at-78 ℃. The reaction was left to melt to room temperature and stirred overnight. Then 40mL of silica gel was added and all volatiles were removed in vacuo. The two regioisomeric products were separated by column chromatography, the desired regioisomer being the product which was first eluted (silica gel, from 85/15 to 80/20 cHex/EE in 30 minutes). 590mg (1.68mmol, 24%) of B-2g and 690mg (1.97mmol, 28%) of the regioisomeric product B-2 g' are separated.

R_f(B-2g) ═ 0.21 (silica gel, cHex: EE 3: 1), [ R-_f(B-2g’)＝0.10]

MS-ESI+：351(M+H)⁺

UV_max＝246nm

3-fluoro-4- (4-methyl- [1.4] diazepan-1-yl) -aniline

2g (12.6mmol) of 3, 4-difluoronitrobenzene are dissolved in 1.6mL of ethanol, 2.4mL (15.1mmol, 1.2eq) of Hunig's base are added and then 1.44g (12.6mmol, 1eq) of hexahydro-1-methyl-1H-1.4-diaza * are added dropwise while cooling with ice. After stirring at room temperature for about 12 hours, the reaction was complete. Methanol and 50mL of silica gel were then added, the volatiles were removed in vacuo and the mixture was purified by column chromatography (DCM/MeOH 97/3 to 85/15 over 35 min). 3g (11.9mmol, 94%) of nitro compound are obtained.

R_f0.39 (silica gel, DCM: MeOH: NH)₃ 9∶1∶0.1)

MS-ESI⁺：253(M+H)⁺

The nitro compound was dissolved in 600mL THF and combined with about 300mg Raney nickel. The mixture was heated at 3bar H₂Hydrogenation under pressure for 3 hours. The raney nickel was filtered off and all volatiles were removed from the solution in vacuo. 2.15g (9.6mmol, 81%) 3-fluoro-4- (4-methyl- [ 1.4) are obtained]Diazepan-1-Yl) -aniline.

R_f0.48 (silica gel, DCM: MeOH: NH)₃ 4∶1∶0.1)

MS-ESI⁺：224(M+H)⁺

The aniline used as educt in example 142-143 was prepared analogously.

4-amino-benzoic acid benzyl ester

10.01g of 4-nitrobenzoic acid are suspended in 500mL of acetonitrile and then combined with 15.03g (108.7mmol, 1.2eq) of potassium carbonate. 15.40g (171.0mmol, 1eq) of benzyl bromide were added dropwise with stirring, and the reaction mixture was then heated for 5 hours at 60 ℃ with stirring. This was combined with 750mL of distilled water, extracted 4 times with 250mL of EE, the organic phases were combined and dried over sodium sulfate. After removal of all volatiles in vacuo, the crude product was then suspended in toluene 2 times and all volatiles were removed in vacuo (removal of excess benzyl bromide). 20.60g (80.1mmol) of benzyl 4-nitro-benzoate were obtained as a colorless solid, which was used in the next step without further purification.

20.6g of benzyl 4-nitro-benzoate were dissolved in 350mL of dioxane and this solution was mixed with 6.9g (49.9mmol, 0.61eq) of Raney nickel. The mixture was heated at 5bar H₂The hydrogenation was carried out under pressure with stirring for 16 hours. The catalyst was filtered off and all volatiles were removed in vacuo. 17.0g (74.8mmol, 93%) benzyl 4-aminobenzoate were obtained as a colorless solid.

C-1a) benzyl 4- (4-chloro-5-trifluoromethyl-pyrimidin-2-ylamino) -benzoate

10g (44mmol) of benzyl 4-aminobenzoate are dissolved in 200mL of DMA, 8mLH u nig of base (0.97eq) are added and 10.4g (48.21mmol) of 2, 4-dichloro-5-trifluoromethylpyrimidine dissolved in 50mL of DMA are added dropwise at room temperature to a clear solution. The reaction solution was stirred at 60 ℃ overnight, then mixed with 300mL of dichloromethane, and extracted with distilled water (3X 300 mL). The organic phase was dried over sodium sulfate and the solvent was removed in vacuo. The crude product was combined with 100mL MeOH, impregnated and left for 2 hours. The mixture was then stirred for 10 minutes and the precipitate was filtered off and washed with methanol (methanol filtrate contains the undesired nucleophilic substitution regioisomer). Finally, the crude product is resuspended in methanol, filtered, washed with a little methanol and dried in a vacuum dryer at 60 ℃. 8.5g (20.7mmol, 43%) of C-1a are obtained in the form of a pale yellow solid.

R_f0.71 (silica gel, cHex: EE 1: 2)

MS-ESI⁺：408(M+H)⁺

C-2a) [4- (4-chloro-5-trifluoromethyl-pyrimidin-2-ylamino) -phenyl ] - (4-methyl-piperazin-1-yl) -methanone

2.74g (6.71mmol) of C-1a were dissolved in 120mL dioxane, 300mg palladium hydroxide (20% w/w Pd, 2.14mmol, 0.32eq) was added, and the mixture was then brought to 3bar H₂Stirred under pressure at room temperature for 16 hours. The reaction mixture was filtered through celite and the solvent removed in vacuo to yield 1.87g (5.89mmol, 88%) of 4- (4-chloro-5-trifluoromethyl-pyrimidin-2-ylamino) -benzoic acid as a colorless solid which was used without further purification. 1.1g (3.46mmol) of benzoic acid were combined with 20mL of toluene and 301. mu.L (4.16mmol, 1.2eq) of thionyl chloride and refluxed for 1.5 hours. All volatiles were removed in vacuo and the crude benzoyl chloride was directly reacted further.

536mg (1.6mmol) of this was dissolved in 4mL of THF and mixed with 410. mu.L (1.5eq) of Hunig base. After the addition of 179. mu.L (1eq) of N-methylpiperazine, the solution was stirred at room temperature for 16 hours. The reaction mixture was poured into about 40mL of distilled water, stirred for 30 minutes, and the aqueous phase was extracted 3 times with 50mL of ethyl acetate. After drying the organic phase over magnesium sulphate, filtration and removal of volatiles in vacuo gave 645mg (1.5mmol, 94%) of C-2a as a solid.

R_f0.69 (silica gel, CH)₂Cl₂∶MeOH∶NH₃ 5∶1∶0.1)

MS-ESI+：400(M+H)⁺

C-2b)4- (4-chloro-5-trifluoromethyl-pyrimidin-2-ylamino) -N-methyl-N- (1-methyl-piperidin-4-yl) -benzamide

R_f0.30 (silica gel, CH)₂Cl₂∶MeOH∶NH₃ 5∶1∶0.1)

MS-ESI+：428(M+H)⁺

C-2b was prepared analogously to C-2a using methyl- (1-methyl-piperidin-4-yl) -amine.

(±)-((1S^*，2R^*) -2-amino-cyclohexyl) -carbamic acid benzyl ester

2mL (16.2mmol) of cis-1, 2-diaminocyclohexane and 2.42g (19.4mmol, 1.2eq) of 9-borabicyclo [3.3.1 ] are added]Nonane (9-BBN) was dissolved in 8mL of THF/NMP 1/1 and stirred at room temperature for 45 minutes. 2.4mL (16.2mmol, 1eq) of benzyl chloroformate (Cbz-chloride) was added to the lightSlightly turbid solutions. After about 1 hour the reaction mixture was combined with distilled water and stirred for a few minutes. The aqueous solution was then combined with ethyl acetate and the aqueous phase was washed 3 times with about 50mL ethyl acetate. The product was completely in the aqueous phase and the impurities in the organic phase. The aqueous phase was made basic with sodium bicarbonate (pH8), mixed with dichloromethane, extracted 3 times with 10mL dichloromethane, the combined organic phases were dried over magnesium sulfate and the solvent was removed in vacuo. 2.29g (9.22mmol, 57%) of (. + -.) ((1S) as a colorless oily liquid were obtained^*，2R^*) -2-amino-cyclohexyl) -carbamic acid benzyl ester.

R_f0.45 (silica gel, CH)₂Cl₂∶MeOH∶NH₃ 9∶1∶0.1)

MS-ESI⁺：249(M+H)⁺

C-3a)(±)-((1S^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclohexyl) -carbamic acid benzyl ester

800mg (2mmol) of C-2a are dissolved in 1mL of NMP, and 569mg (2.4mmol, 1.2eq) (+ -) - ((1S) are added^*，2R^*) -2-amino-cyclohexyl) -carbamic acid benzyl ester, then 521 μ L (3mmol, 1.5eq) of Hunig's base is added. After heating at 70 ℃ for 48 hours, the reaction had stopped. After removal of the solvent in vacuo, the crude product is purified by column chromatography (DCM/MeOH/NH)₃From 19/1/0.1 to 9/1/0.1), 826mg (1.35mmol, 68%) of product are obtained in the form of a colorless resin.

MS-ESI⁺：612(M+H)⁺

C-3b)(±)-{4-[4-((1R^*，2S^*) -2-amino-cyclohexylamino) -5-trifluoromethyl-pyrimidin-2-ylamino]-phenyl } - (4-methyl-piperazin-1-yl) -methanone

112mg (0.18mmol) of C-3a were dissolved in DMF (10mL) and combined with distilled water (1 mL). Then 9mL of DMF was added and the solution was transferred to the hydrogenation unit and combined with Pd/C (200mg, 5% Pd). The reaction solution was heated at 4bar H₂Stirred under pressure for 12 hours. The reaction mixture was taken up in dichloromethane, combined with 10mL of RP-gel and all volatiles were removed in vacuo. Purification was done by column chromatography (RP-phase, acetonitrile/water from 5/95 to 95/5 in 20 min). After combining the product fractions and freeze-drying, 27mg (0.06mmol, 30%) of the desired product are obtained as a colorless solid.

MS-ESI⁺：478(M+H)⁺

C-3c)(±)-(1S^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cycloheptanecarboxylic acid

440mg (1.1mmol) of C-2a are dissolved in 500. mu.L of NMP and combined with 565. mu. L H ug of base (3.3mmol, 3eq) and 256mg of cis-2-aminocycloheptane carboxylic acid (racemic). The reaction mixture was placed in an oil bath maintained at 100 ℃ and heated with stirring to this temperature for 8 hours. After the reaction was complete, the reaction mixture was taken up in methanol, combined with 20mL of RP-gel and all volatiles were removed in vacuo. Purification is done by reverse phase (eluent: acetonitrile/water (15/85 to 35/65 in 15 min.) after combining the product components and freeze drying, 160mg (0.31mmol, 28%) of the desired product are obtained as a colorless solid.

MS-ESI⁺：521(M+H)⁺

C-3d)(±)-(1S^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclopentanecarboxylic acid

563mg (1.13mmol) of C-2a was dissolved in 5mL of 1-butanol, and 163mg of cis-2-amino-1-cyclopentanecarboxylic acid (racemic) was added thereto. After addition of 540. mu.L of Hunig's base, the mixture was heated to 110 ℃ for about 60 minutes (microwave, CEM, 100W). The reaction mixture was evaporated to dryness in vacuo, stirred with about 100mL of water and extracted 3 times with 50mL of ethyl acetate. The combined organic phases were dried over magnesium sulfate and the solvent was removed in vacuo. 530mg (1.08mmol, 96%) of C-3d are obtained.

MS-ESI⁺：493(M+H)⁺

C-3e was prepared analogously using DMA as solvent and C-2b as starting material.

MS-ESI⁺：521(M+H)⁺

C-3f) (1S, 3R) -3- (2- {4- [ methyl- (1-methyl-piperidin-4-yl) -carbamoyl ] -phenylamino } -5-trifluoromethyl-pyrimidin-4-ylamino) -cyclopentanecarboxylic acid

200mg of C-2b were dissolved in 750. mu.L of DMA and 160. mu.L (0.93mmol, 2eq) of Hunig base were added. Then 72mg (0.56mmol, 1.2eq) of (1S, 3R) -3-aminocyclopentanecarboxylic acid were added and the reaction mixture was heated at 120 ℃ for 40 min. The reaction mixture was combined with RP-gel, volatiles removed in vacuo, and the product purified by RP-phase column chromatography (from 85% water (+ 0.2% formic acid) and 15% acetonitrile (+ 0.2% formic acid) to 76% water and 24% acetonitrile over 20 min). The corresponding product fractions were combined and lyophilized to remove the solvent, yielding 150mg (0.29mmol, 62%) of C-3f as a colorless film.

(±) -trans-2-aminocyclopentanecarboxamide

The compound was prepared according to the literature (Csomos et al, 2002).

D-2a) benzyl 4- [4- ((1R, 2S) -2-carboxy-cyclopentylamino) -5-trifluoromethyl-pyrimidin-2-ylamino ] -benzoate

2.05g (5mmol) of C-1a and 1g of (1S, 2R) - (+) -2-amino-1-cyclopentane carboxylate (6mmol, 1.2eq) were placed in 18mL of ethanol. 7.3ml (42.5mmol, 3.4eq) of Hunig base are added and the mixture is stirred at 70 ℃ for 4 hours. The reaction mixture was stirred into 275mL of water, the insoluble material was removed by filtration, and the filtrate was saturated with KHSO₄The aqueous solution was adjusted to pH2, stirred for 5 minutes and the precipitate formed was filtered off with suction. The crude product was washed with water and dried in vacuo to yield 2.37g (4.74mmol, 94%) of D-2a as a pale brown solid.

MS-ESI⁺：501(M+H)⁺

Analogously to obtain (1R, 2S) - (-) -2-amino-1-cyclopentanecarboxylic acid or (1R)^*，2S^*) Synthesis of (- + -) -2-amino-1-cyclopentanecarboxylic acid derivative. The corresponding products were designated D-2b (chiral enantiomer of D-2a) and D-2c (rac).

Preparation of (1S, 2R) -2-aminocyclopentanecarboxylic acid hydrochloride

22.64mL (0.26mol, 0.95eq) of CSI was added dropwise to 23mL (0.273mol, 1eq) of cyclopentene under argon at-75 ℃. The reaction temperature is kept below-65 ℃ all the time when the catalyst is added. The reaction was allowed to reach room temperature over 2 hours and was further stirred overnight. The work-up after reduction was carried out by adding the reaction solution dropwise to 600mL of an ice/water solution with 60g of sodium sulfite and 180g of sodium bicarbonate. The aqueous phase is extracted 4 times with 200mL of dichloromethane, the organic phases are combined, dried over magnesium sulfate and all volatiles are removed in vacuo. 25.75g (85%) of pale yellow crystals were obtained.

These were dissolved in 400mL of diisopropyl ether, 1.6mL of water and 20g of resin-bonded lipolase (lipase acrylic resin from Candida antartica, Sigma-Aldrich) were added, and the mixture was stirred at 60 ℃ for 11 days. The reaction suspension was filtered through celite, washed with diisopropyl ether and the filtrate was evaporated to dryness. The resulting pale yellow oil was dissolved in 200mL of dichloromethane and washed with about 150mL of saturated sodium bicarbonate solution. The aqueous phase was extracted 3 times with dichloromethane and the organic phases were combined and dried over magnesium sulphate. After removal of all volatiles in vacuo, 8.93g of chiral lactam were obtained as a pale yellow oil.

The latter product was dissolved in 10mL of water and 10mL of 37% HCl (aq) was added with stirring while cooling in an ice bath. After stirring at 0 ℃ for 10 minutes, the reaction solution was left to stand at room temperature overnight. The precipitated crystals are filtered off, washed with a small amount of acetonitrile and dried under high vacuum. The mother liquor was evaporated to almost dryness and the precipitated crystals were filtered off, washed with acetonitrile and also dried under high vacuum. 11.74g (70.9mmol, 31% based on racemic lactam) of (1S, 2R) -2-aminocyclopentanecarboxylic acid hydrochloride were obtained as colorless crystals (the enantiomeric acid had precipitated and contained in the precipitate in the kinetic resolution step, isolated by filtration through celite).

The synthesis sequence is described in the literature (Forro and Fueloep, 2003).

D-3a) benzyl 4- [4- ((1R, 2S) -2-isopropylcarbamoyl-cyclopentylamino) -5-trifluoromethyl-pyrimidin-2-ylamino ] -benzoate

2.59g (4.9mmol) D-2a, 2.21g (6.9mmol, 1.4eq) TBTU and 4.21mL (24.6mmol, 5eq) Hunig's base were dissolved in 75mL DMF and stirred at room temperature for 20 min. Then 0.63ml (7.38mmol, 1.5eq) of isopropylamine were added and the mixture was stirred at room temperature overnight. It is filtered through basic alumina, washed with DMF, the mother liquor is stirred into 400mL of water, stirred for 30 min and the precipitate is filtered through suction. The crude product was washed with water and dried in vacuo. For purification, it is stirred with 50ml of acetonitrile at 5 ℃ for 30 minutes, filtered off with suction, washed with some cold acetonitrile and the residue is dried in vacuo. 2.13g (3.9mmol, 80%) of D-3a are obtained in the form of a light brown solid.

R_f0.53 (silica gel, cHx: EE 1: 1)

MS-ESI⁺：542(M+H)⁺

D-4a)4- [4- ((1R, 2S) -2-isopropylcarbamoyl-cyclopentylamino) -5-trifluoromethyl-pyrimidin-2-ylamino ] -benzoic acid

2.13g (3.9mmol) of D-3a are dissolved in 150mL of THF, and 250mg of palladium hydroxide/C-catalyst (20% by weight of Pd on charcoal) are added. The mixture was cooled to room temperature under 6 bar H₂The hydrogenation was carried out under pressure with stirring for 16 hours. Then 30mL of methanol were added, the catalyst was filtered through celite, washed with methanol and the filtrate was evaporated to dryness. The residue was boiled in 45mL of ethanol, slowly cooled to 5 ℃, stirred for a further 1 hour, then filtered off with suction and washed with cold ethanol. 2.46g (3.2mmol, 82%) of the acid D-4a are obtained.

R_f0.46 (silica gel, CH)₂Cl₂∶MeOH∶AcOH 5∶1∶0.1)

MS-ESI⁺：452(M+H)⁺

Enantiomeric compounds and racemic compounds were synthesized analogously.

D-5c)(±)-{4-[4-((1R^*，2S^*) -2-isopropylcarbamoyl-cyclopentylamino) -5-trifluoromethyl-pyrimidin-2-ylamino]-phenyl } -carbamic acid tert-butyl ester

450mg (1mmol) of D-4c are dissolved in 1.8mL of dry toluene and 222. mu.L (1.3mmol, 1.3eq) of Hunig base and 940. mu.L of tert-butanol are added successively. Then 258. mu.L of diphenylphosphoryl azide was added and the mixture was heated at 80 ℃ for 16 hours. The reaction mixture was combined with 20mL of ethyl acetate, washed 2 times with 20mL of 0.5M NaOH solution, and the aqueous phase was back-washed 2 times with 20mL of ethyl acetate. The combined organic phases were washed with saturated aqueous sodium chloride solution, the insoluble constituents were filtered off, the filtrate was dried over magnesium chloride and the solvent was removed in vacuo. 461mg (0.88mmol, 89%) of D-5c are obtained in the form of a pale yellow solid.

MS-ESI⁺：523(M+H)⁺

D-6c)(±)-(1S^*，2R^*) -2- [2- (4-amino-phenylamino) -5-trifluoromethyl-pyrimidin-4-ylamino]-cyclopentanecarboxylic acid isopropylamide

461mg (0.88mmol) of D-5c were dissolved in 5mL of dichloromethane, 2mL of trifluoroacetic acid was added, and the mixture was stirred at room temperature for 1 hour. The reaction mixture was stirred into 50mL of water and the aqueous phase was washed with 50mL of ethyl acetate. The organic phases are extracted 2 more times with 30mL of 10% hydrochloric acid, the aqueous phases are combined, adjusted to pH10 with 10% sodium hydroxide solution and extracted 3 more times with 50mL of ethyl acetate. The combined organic phases were dried over magnesium sulphate and the volatiles were removed in vacuo to yield 243mg (0.58mmol, 65%) of D-6c as a colourless solid.

R_f0.08 (silica gel, cHex: EE 1: 1)

MS-ESI⁺：423(M+H)⁺

E-1) 2-methylthio-1H-pyrimidin-4-one

20g (153mmol) of 2-thiouracil are suspended in 250mL of methanol, and 8.7g (152.9mmol, 1eq) of sodium methoxide are added. The solution was stirred at room temperature for 5 minutes, then 12.4mL (198.8mmol, 1.3eq) of iodomethane was added dropwise. The reaction mixture was stirred overnight, then poured onto water and extracted 3 times with about 150ml chloroform. The combined organic phases were dried over magnesium sulfate and the solvent was removed in vacuo to yield 16g (121.5mmol, 74%) of E-1 as a colorless solid.

E-2)4- (6-oxo-1, 6-dihydro-pyrimidin-2-ylamino) -benzoic acid

4.1g (28.8mmol) of E-1 are dissolved in 10mL of diglyme (diglyme) and the solution is combined with 4.79g (34.6mmol, 1.2eq) of 4-aminobenzoic acid. The reaction mixture was refluxed for 16 hours. After cooling to room temperature, the precipitate is filtered off with suction, washed with a small amount of diglyme, then with diethyl ether and dried in vacuo. 5.27g (22.8mmol, 79%) of E-2 are obtained as a colorless solid.

MS-ESI⁺：232(M+H)⁺

E-3a)4- (5-iodo-6-oxo-1, 6-dihydro-pyrimidin-2-ylamino) -benzoic acid

9g (38.9mmol) of E-2 were placed in 100mL of water, and 2.18g of NaOH (54.5mmol, 1.4eq) were added. The solution was combined with 11.9g (46.7mol, 1.2eq) iodine and stirred at 65 ℃ for 3 h. After cooling to 50 ℃, sodium thiosulfate pentahydrate was added to remove excess iodine, and then the mixture was stirred for another 1 hour and cooled to room temperature. The brown precipitate was filtered off with suction, washed with water and dried in vacuo. 13.7g (38.4mmol, 82%) of E-3a are obtained.

MS-ESI⁺：358(M+H)⁺

E-3b)4- (5-bromo-6-oxo-1, 6-dihydro-pyrimidin-2-ylamino) -benzoic acid

9g (38.9mmol) of E-2 was placed in 10mL of acetic acid, a 2.1mL (40.9mmol, 1.05eq) solution of bromine dissolved in 50mL of acetic acid was added dropwise thereto, and the mixture was stirred at room temperature for about 1 hour. The reaction mixture is stirred into 800mL of water, the precipitate is filtered off with suction, the brown precipitate obtained is washed with water and dried in vacuo. 11.5g (37.1mmol, 95%) of E-3b are obtained as a colorless solid.

R_f0.27 (silica gel, EE: MeOH 7: 3)

MS-ESI⁺：309/311(M+H)⁺(1x Br)

E-4a)4- (4-chloro-5-iodo-pyrimidin-2-ylamino) -benzoyl chloride and

e-5a)4- (4-chloro-5-iodo-pyrimidin-2-ylamino) -benzoic acid

6.5g (18.2mmol) of E-3a are suspended in 80mL of phosphorus oxychloride and the mixture is stirred under reflux for 3 hours. The reaction mixture was added dropwise to 800mL of water/ice with vigorous stirring and stirred for a further 30 minutes, the crude acid chloride E-4a was filtered off. It was dried in vacuo and used further without any purification. To prepare this acid, the crude acid chloride was dissolved in 200mL THF and 200mL 20% aqueous sodium bicarbonate was added. The reaction mixture was stirred at room temperature for 16 hours. The THF is removed in vacuo, the aqueous phase is adjusted to pH2 with concentrated HCl, stirred for 10 minutes, and the residue formed is filtered off with suction and washed with water. After drying in vacuo, 6.3g (16.7mmol, 92%) of E-5a are obtained as a colourless solid.

R_fNot 0.24 (silica gel, ethyl acetate)

MS-ESI⁺：427(M+H)⁺

E-4b)4- (4-chloro-5-bromo-pyrimidin-2-ylamino) -benzoyl chloride and

e-5b)4- (4-chloro-5-bromo-pyrimidin-2-ylamino) -benzoic acid

Derivatives E-4a and E-5a were prepared analogously from E-3 b.

E-6b) [4- (5-bromo-4-chloro-pyrimidin-2-ylamino) -phenyl ] - (4-methyl-piperazin-1-yl) -methanone

559mg (1.6mmol) of E-4b are dissolved in 5mL of THF and 414. mu.L (2.4mmol, 1.5eq) of Hunig base are mixed. mu.L (1.6mmol, 1eq) of N-methylpiperazine was added dropwise to this solution, and the mixture was stirred at room temperature for 90 minutes. Then 100mL of water were added and the mixture was extracted 3 times with 50mL of ethyl acetate. The combined organic phases were dried over magnesium sulfate and the solvent was removed in vacuo. 566mg (1.4mmol, 86%) of E-6b are obtained in the form of a colorless resin.

MS-ESI⁺：410/412(M+H)⁺(1x Br)

E-7b)(±)-(1S^*，2R^*) -2- { 5-bromo-2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-pyrimidin-4-ylamino } -cyclopentanecarboxylic acids

459mg (1.1mmol) of E-6b are dissolved in 5mL of 1-butanol and 536. mu.L (3.1mmol, 2.8eq) of Hunig base are combined. 162mg of cis-2-aminocyclopentanecarboxylic acid (racemic) were added to the solution, and the reaction mixture was stirred at 110 deg.C (CEM microwave, 100W) for 100 minutes. The reaction mixture was evaporated to dryness, added to about 200mL of water with stirring, and extracted 3 times with 50mL of ethyl acetate. The combined organic phases were dried over magnesium sulfate and the solvent was removed in vacuo. 321mg (0.64mmol, 57%) of E-7b are obtained in the form of a colorless resin.

MS-ESI⁺：503/505(M+H)⁺(1x Br)

E-8b) (±) -4- [ 5-bromo-4- ((1R)^*，2S^*) -2-carbamoyl-cyclopentylamino) -pyrimidin-2-ylamino]-benzoic acid

1g (3.04mmol) of E-5b are dissolved in 3.9mL of DMA and 1.3. mu.L (7.6mmol, 1.5eq) of Hunig base are combined. 390mg (3.04mmol, 1eq) of cis-2-aminocyclopentanecarboxamide (racemic) are added to the solution and the reaction mixture is stirred for 60 minutes at 120 ℃. The reaction mixture was evaporated to dryness and the residue was treated with 5ml of 1-butanol and the precipitate was filtered off with suction. After washing with 5mL of cold 1-butanol, it was dried in vacuo to give 935mg (2.2mmol, 73%) of E-8b as a pale brown solid.

MS-ESI⁺：420/422(M+H)⁺(1x Br)

The iodo derivative E-8a was prepared analogously from E-5 a. However, the reaction temperature was 80 ℃.

E-9b)(±)-4-[4-((1R^*，2S^*) -2-carbamoyl-cyclopentylamino) -5-cyano-pyrimidin-2-ylamino]-benzoic acid

935mg (2.23mmol) of E-8b was dissolved in 8mL of DMF and 403mg (4.45mmol, 2eq) of copper (I) cyanide was added under argon. The yellow solution is combined with 80mg (0.067mmol, 3 mol%) of palladium-tetrakis (triphenylphosphine) and heated at 145 ℃ for 24 hours, during which about 50% of the educts react. The same amount of catalyst was added, the mixture was heated for a further 5 hours and the reaction was worked up. The reaction mixture was filtered through a glass vessel filled with silica gel (solvent: DMF), and the filtrate was evaporated to about 5mL and poured into about 400mL of distilled water. The precipitate formed was filtered off, washed with 100mL of water and dissolved in methanol. RP-gel was added and the solvent was removed in vacuo. The mixture was purified by reverse phase chromatography (from 5% acetonitrile (+ 0.2% formic acid) to 95% water (+ 0.2% formic acid) to 50% acetonitrile (+ 0.2% formic acid) and 50% water (+ 0.2% formic acid)). 160mg (0.44mmol, 20%) of a beige solid E-9b are isolated.

R_f0.30 (silica gel, CH)₂Cl₂∶MeOH∶AcOH 5∶1∶0.1)

MS-ESI⁺：367(M+H)⁺

Example 1

(±)-(1S^*，2R^*) -2- {2- [4- (methyl-phenyl-sulfamoyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclopentanecarboxamide (synthesis scheme A)

150mg (0.6mmol) A-2, 519mg (1.98mmol, 3eq) 4-amino-N-methyl-N-phenyl-benzenesulfonamide and 130. mu.L (0.76mmol, 1.15eq) N-ethyldiisopropylamine are dissolved in 3mL N, N-dimethylacetamide and the solution is stirred at 180 ℃ for 10 min (heating in a microwave). The solution was stirred into 30mL of water, adjusted to pH3 with 0.1N HCl (aq), extracted 3 times with 10mL of ethyl acetate, dried over magnesium sulfate and the volatiles removed in vacuo. The residue was purified by column chromatography (cyclohexane/ethyl acetate 2/1). 92mg (0.2mmol) of N-methyl-4- (4-methylsulfanyl-5-trifluoromethyl-pyrimidin-2-ylamino) -N-phenyl-benzenesulfonamide are obtained as a light brown solid.

85mg (0.19mmol) of this intermediate are dissolved in 7.5mL of dichloromethane, 64mg (0.285mmol, 1.5eq, 77%) of m-chloroperoxybenzoic acid are added and the mixture is stirred at room temperature for 3 hours. The organic phase is washed 3 times with 20ml of saturated aqueous sodium bicarbonate solution, whereby 3-chlorobenzoic acid is removed. After drying the organic phase over sodium sulfate, 83mg (0.18mmol, 95%) of 4- (4-methanesulfinyl-5-trifluoromethyl-pyrimidin-2-ylamino) -N-methyl-N-phenyl-benzenesulfonamide (A-4a) are obtained and are used in the next step without further purification.

83mg (0.18mmol) of A-4a, 26mg of cis-2-amino-1-cyclopentanecarboxamide (0.2mmol, 1.1eq, racemic) and 35. mu.L (0.2mmol, 1.1eq) of Hunig base are dissolved in 2mL of DMA and stirred at 60 ℃ for 1H. The reaction mixture was stirred into 10mL of 0.1N HCl (aq), the mixture was stirred for 30 minutes, and the precipitate formed was filtered off with suction, washed with water and dried. Finally, purification was accomplished by column chromatography (cHex/EE 60/40 to 50/50 in 20 min). 43mg (0.08mmol, 45%) of compound 1 are obtained as a colorless solid.

Examples 2 and 3 were prepared similarly.

Example 4

(±)-N-((1S^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclohexyl) -acetamide (synthesis scheme C)

38mg (0.08mmol) of C-3b are dissolved in 50. mu.L of DMA, 25. mu.L (0.16mol, 2eq) of Hunig base are added and dissolved at room temperature for a few minutes. mu.L of acetyl chloride (1eq) was dissolved in a small amount of DMA and added dropwise to the reaction mixture. After about 10 minutes, the reaction mixture was taken up in dichloromethane, combined with 10mL RP-gel and all volatiles were removed in vacuo. The mixture was purified by RP chromatography (AcCN/water changed from 5/95 to 95/5% in 20 min). The product fractions were combined and lyophilized to give 18mg (0.034mmol, 42%) of compound 4 as a colorless solid.

Examples 5-12 were prepared similarly.

Example 13

(±) -1-methyl-3- ((1S)^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclohexyl) -urea (synthesis scheme C)

50mg (0.105mmol) of C-3b are dissolved in 50. mu.L of DMF and combined with 55. mu.L (0.315mmol, 3eq) of Hunig base. To this solution 6. mu.L of methyl isocyanate (1eq) was added at room temperature. After about 10 minutes, the reaction mixture was taken up in dichloromethane, combined with 10mL of RP-gel and all volatiles were removed in vacuo. The mixture was purified by RP chromatography (change of AcCN/water from 5/95 to 95/5% in 20 min). The product fractions were combined and lyophilized to give 24mg (0.045mmol, 43%) of compound 13 as a colorless solid.

Examples 14-17 were prepared similarly.

Example 18

((±)-(1S^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclohexyl) -carbamic acid methyl ester (synthesis scheme C)

30mg (0.063mmol) of C-3b are dissolved in 50. mu.L of DMF and combined with 22. mu.L (0.126mmol, 2eq) of Hunig base. mu.L of methyl chloroformate (1.2eq) was added to this solution at room temperature. After about 10 minutes, the reaction mixture was taken up in dichloromethane, combined with 10mL of RP-gel and all volatiles were removed in vacuo. The mixture was purified by RP chromatography (AcCN/water changed from 5/95 to 95/5% in 20 min). The product fractions have been combined and lyophilized to give 13mg (0.025mmol, 39%) of compound 13 as a colorless solid.

Examples 19 and 20 were prepared similarly.

Example 21

[4- (4-Cyclopentylamino-5-trifluoromethyl-pyrimidin-2-ylamino) -phenyl ] - (4-methyl-piperazin-1-yl) -methanone (FIG. C)

88mg (0.22mmol) of C-2a are dissolved in 290. mu.L of DMA, 26. mu.L (0.26mmol, 1.2eq) of cyclopentylamine and 75. mu.L (0.44mmol, 2eq) of Hunig base are added and the reaction mixture is heated to 120 ℃. After about 90 minutes, the reaction mixture was poured into about 10mL of distilled water and the precipitate formed was filtered off. The suspension is extracted 3 times with 20mL of ethyl acetate, the combined organic phases are dried over saturated aqueous NaCl solution and magnesium sulfate, combined with 100 μ L of dioxane HCl and all volatiles are removed in vacuo. 106mg (0.219mmol, 99%) of compound 21 are obtained as the hydrochloride salt.

Examples 22-26 were prepared similarly.

Example 27

(±)-(1S^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cycloheptanecarboxylic acid dimethylamide (FIG. C)

35mg (0.067mmol) of C-3d are dissolved in 250. mu.L of DMF, 30. mu.L (0.175mmol, 2.6eq) of Hunig base are added, followed by 35mg (0.11mmol, 1.6eq) of TBTU. The reaction mixture was stirred at room temperature for 10 min, then combined with 118 μ L dimethylamine (2M in THF, 0.235mmol, 3.5 eq). The mixture was stirred at 35 ℃ for 4 hours, then the reaction mixture was taken up in acetonitrile, combined with 6mL RP-gel and all volatiles were removed in vacuo. Purification was accomplished by RP column chromatography (acetonitrile/water change from 12/88 to 40/60 in 12 minutes). The product fractions were lyophilized to give 19mg (0.035mmol, 52%) of compound 27.

Examples 28-30 were prepared similarly.

Example 31

(±)-4-[4-((1R^*，2S^*) -2-isopropylcarbamoyl-cyclopentylamino) -5-trifluoromethyl-pyrimidin-2-ylamino]-N- [2- (1-methyl-pyrrolidin-2-yl) -ethyl]Benzamide (synthesis scheme D)

80mg (0.18mmol) of D-4c are dissolved in 2.4mL of DMF, 179. mu.L (1.03mol, 1.5eq) of Hunig base are added and the solution is combined with 83mg (0.25mmol, 1.4eq) of TBTU. The solution was stirred at room temperature for 40 minutes, then 38.5. mu.L (0.27mmol, 1.5eq) of 2- (2-aminoethyl) -1-methylpyrrolidine was added and the mixture was stirred for 2 days. Silica gel was then added to the reaction mixture and the volatiles were removed in vacuo. Purification was accomplished by normal phase chromatography using column chromatography (DCM/MeOH/NH)₃(aq) 5/1/0.1). 70mg (0.125mmol, 70%) of compound 31 are obtained.

Examples 32-58 were prepared similarly.

Example 59

(±)-(1S^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclopentanecarboxylic acid isopropylamide (FIG. C)

88mg (0.18mmol) of C-3d are dissolved in 2mL of DMF, 153. mu.L (0.90mmol, 5eq) of Hunig base are added and the solution is combined with 81mg (0.25mmol, 1.4eq) of TBTU. The solution was stirred at room temperature for 20 minutes, then 12. mu.L (0.27mmol, 1.5eq) of isopropylamine were added and the mixture was stirred for 16 hours. It was then filtered through basic alumina and washed with 20mL of methanol. The RP gel was added to the filtrate and the volatiles were removed in vacuo. The crude product immobilized on RP-gel was purified by reverse phase (changing from 95% water (+ 0.2% formic acid) and 5% acetonitrile (+ 0.2% formic acid) to 55% water and 45% acetonitrile in 20 minutes). The corresponding product fractions were combined with 1eq concentrated hydrochloric acid and lyophilized to remove the solvent. 14mg (0.025mmol, 14%) of compound 59 as a colorless film are left as the hydrochloride salt.

Examples 60-69 were prepared similarly.

Examples 68 and 69 were chiral and were prepared accordingly from C-2a using the enantiomer of cis-2-aminocyclopentanecarboxylic acid followed by the formation of the prepared isopropylamide. Alternatively, 68 and 69 can also be obtained from 59 by preparative chiral HPLC.

Example 70

(±)-(1S^*，2R^*) -2- {2- [ 3-chloro-4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclopentanecarboxylic acid isopropylamide (FIG. B)

30mg (85.5mmol) of B-2a are dissolved in 100. mu.L of NMP and combined with 35mg (0.14mmol, 1.6eq) (4-amino-2-chloro-phenyl) - (4-methyl-piperazin-1-yl) -methanone. To this reaction mixture was added 107 μ L of 4M HCl in dioxane (0.43mmol, 5eq) which was stirred at 5 ℃ for 12 hours. The reaction mixture was taken up in DCM/MeOH/NH₃9/1/0.1, combined with 6mL of RP-gel, and the volatiles removed in vacuo and purified by RP chromatography (from 5% acetonitrile to 95% acetonitrile over 10 min). The corresponding product components were lyophilized to remove the solvent. Residue 35mg (0.06mmol, 72%) of Compound 70.

Examples 71-75 were prepared similarly.

Examples 76 to 105 (general method)

1eq of compound B-4 (E-8B for examples 98-101 and E-8a for example 102-105) was dissolved in DMF (ca. 1-10mL/mmol), 4-6eq Hunig base was added, followed by 1.3-1.5eq TBTU. The reaction mixture was stirred at room temperature for 10-30 minutes, then 1-1.5eq amine or aniline were added. After the reaction is complete, the reaction mixture is combined with silica gel, all volatiles are removed in vacuo, and the product is purified by column chromatography (normal phase or RP-phase) and isolated.

Example 106

(±)-(3-[4-((1R^*，2S^*) -2-carbamoyl-cyclopentylamino) -5-trifluoromethyl-pyrimidin-2-ylamino]-N-phenylbenzamides (scheme A)

700mg (3.06mmol) of A-3 was dissolved in 6mL of DMA. 800. mu.L (4.6mmol, 1.5eq) of Hunig base were added and 440mg of cis-2-amino-1-cyclopentanecarboxamide dissolved in 24mL of DMA were added dropwise. The reaction mixture was stirred at room temperature. After 1 hour it was diluted with 400mL dichloromethane, extracted 2 times with 200mL half-saturated ammonium chloride solution, then dried over magnesium sulfate and the solvent removed in vacuo. 1.1g of crude (. + -.) - (1S) remained^*，2R^*) -2- (2-methylsulfanyl-5-trifluoromethyl-pyrimidin-4-ylamino) -cyclopentanecarboxamide as a beige solid. It was reacted further without purification.

For this purpose, this solid is dissolved in 60mL of THF, 1.31g (5.5mmol, 77% 2eq) of mCPBA are added portionwise, and the mixture is stirred at room temperature for 1 hour. The organic phase is washed 3 times with 20ml of saturated aqueous sodium bicarbonate solution, whereby 3-chlorobenzoic acid is removed. After drying the organic phase over magnesium sulfate, 1.15g of crude (. + -.) - (1S) were obtained^*，2R^*) -2- (2-methanesulfinyl-5-trifluoromethyl-pyrimidin-4-ylamino) -cyclopentane carboxamide, which was used in the next step without further purification.

150mg (0.45mmol) (+ -) - (1S)^*，2R^*) -2- (2-methanesulfinyl-5-trifluoromethyl-pyrimidin-4-ylamino) -cyclopentanecarboxamide dissolved in 500. mu.l NMP and 148mg (0.68mmol, 1.5eq) of m-aminobenzanilide (m-amino-benzanilide) added. To this solution was added 34 μ L of hydrochloric acid (4M in dioxane, 0.3eq) which was stirred at 50 ℃ for 16 hours. The reaction mixture was stirred into 30mL of water, adjusted to pH3 with 10mL of 0.1N HCl, and extracted 3 times with 15mL of ethyl acetate. The combined organic phases were dried over magnesium sulphate, all volatiles were removed in vacuo, the crude product was stirred into cyclohexane/ethyl acetate 60/40, and the precipitate was filtered off with suction and washed with 2-propanol. 15mg (0.03mmol, 7%) of compound 106 are obtained as a colorless solid.

Example 107-109 was prepared similarly. Here, purification is done by column chromatography (ethyl acetate/cyclohexane, silica gel).

Example 110

(±) ((1S, 2R) -2- { 5-bromo-2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino ] -pyrimidin-4-ylamino } -cyclopentanecarboxylic acid cyclopropylamide (FIG. E)

39mg (0.077mmol) of E-7b are dissolved in 500. mu.L of DMF, and 66. mu.L (0.39mmol, 5eq) of Hunig base and 35mg (0.11mmol, 1.4eq) of TBTU are added. The solution was stirred at room temperature for 20 minutes, then 8. mu.L (0.116mmol, 1.5eq) of cyclopropylamine were added and the mixture was left at room temperature overnight. It was filtered through basic alumina, washed with about 20mL of methanol, and the filtrate was combined with 8mL of RP-gel. After removal of volatiles in vacuo, the mixture was purified by reverse phase (changing from 95% water (+ 0.2% formic acid) and 5% acetonitrile (+ 0.2% formic acid) to 5% water and 95% acetonitrile over 20 minutes). The corresponding product components were lyophilized to remove the solvent. 12mg (0.021mmol, 27%) of compound 110 are obtained as a colorless thin film.

MS-ESI⁺：542/544(M+H)⁺(1Br)

Example 111-120 was prepared similarly.

Example 121

N-methyl-N- (1-methyl-piperidin-4-yl) -4- {4- [ (±) - (1R^*，2S^*) -2- (pyrrolidine-1-carbonyl) -cyclopentylamino]-5-trifluoromethyl-pyrimidin-2-ylamino } -benzamide (synthesis scheme C)

80mg (0.15mmol) of C-3e are dissolved in 1.4mL of DMF and 132. mu.L (0.77mmol, 5eq) of Hunig base and 69mg (0.22mmol, 1.4eq) of TBTU are added. The reaction mixture was stirred at room temperature for 30 minutes, then 119. mu.L (0.144mmol, 9.4eq) of pyrrolidine was added and the mixture was stirred at room temperature for 16 hours. It was filtered through basic alumina, washed with about 20mL of methanol, and the filtrate was combined with silica gel. After removal of volatiles in vacuo, the mixture was purified by column chromatography (DCM/MeOH/NH)₃9/1/0.1). The product fractions were collected and combined with 100 μ L HCl (4M in dioxane) and the solvent was removed in vacuo to yield 29mg (0.048mmol, 31%) of compound 121 as the hydrochloride salt as a colorless film.

MS-ESI⁺：574(M+H)⁺

Example 122-128 was similarly prepared.

Example 129

4- [4- ((1R, 3S) -3-carbamoyl-cyclopentylamino) -5-trifluoromethyl-pyrimidin-2-ylamino ] -N-methyl-N- (1-methyl-piperidin-4-yl) -benzamide (FIG. C)

75mg (0.14mmol) of C-3f are dissolved in 1mL of DMF, 123. mu.l (0.7mmol, 5eq) of Hunig's base are added and the reaction mixture is stirred for 30 minutes. Then 14. mu.L (0.22mmol, 1.5eq) of aqueous ammonia solution (28%) was added and the mixture was stirred at room temperature for 5 hours. The solution was combined with RP-gel, all volatiles were removed in vacuo, and the mixture was purified by column chromatography (changing from 10% acetonitrile (+ 0.2% formic acid) and 90% water (+ 0.2% formic acid) to 24% acetonitrile and 76% water over 12 minutes). The product fractions were combined with 100. mu.L dioxane HCl and lyophilized to remove all volatiles. 35mg (0.063mol, 44%) of compound 129 are obtained in the form of the hydrochloride.

Example 130 was prepared similarly.

Example 131

(±)-(1S^*，2R^*) -2- [2- (4-acetylamino-phenylamino) -5-trifluoromethyl-pyrimidin-4-ylamino]Isopropylamide cyclopentanecarboxylic acid (FIG. D)

22mg of D-6c are dissolved in 1mL of THF, combined with 14. mu.L (0.075mmol, 1.5eq) of Hunig's base, and then 3. mu.L of acetyl chloride dissolved in 500. mu.L of LTHF is added. After about 90 minutes the reaction solution was diluted with 10mL of methanol and 8mL of RP-gel was added. Chromatographic purification was done by reverse phase (changing from 78% water (+ 0.2% formic acid) and 22% acetonitrile (+ 0.2% formic acid) to 51% water and 49% acetonitrile over 15 minutes). The corresponding product components were combined and lyophilized to remove the solvent. 14mg (0.028mmol, 54%) of compound 131 are obtained.

Example 132-133 was prepared similarly.

Example 134

(±)-(1S^*，2R^*) -2- { 5-cyano-2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-pyrimidin-4-ylamino } -cyclopentanecarboxamide (scheme E)

40mg (0.11mmol) of E-9b are dissolved in 1.5mL of DMF, 110. mu.L (0.63mmol, 5.8eq) of Hunig base are added and the reaction mixture is stirred for 40 minutes. Then 18. mu.L (0.16mmol, 1.5eq) of N-methylpiperazine were added and the mixture was stirred at room temperature for 48 hours. The solution was combined with silica gel, all volatiles were removed in vacuo, and the mixture was purified by column chromatography (DCM/MeOH 9/1). 33mg (0.07mol, 67%) of compound 134 are obtained.

Example 135-136 was prepared similarly.

Example 137

(±)-(1S^*，2R^*) -2- { 5-Cyclopropylethynyl-2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-pyrimidin-4-ylamino } -cyclopentanecarboxamide (scheme E)

50mg (0.09mmol) of 105 are dissolved in 220. mu.L of DMF,then 15mg of dichloro-bis (triphenylphosphine) palladium (0.021mmol, 23 mol%) and 10mg (0.03mmol, 0.58eq) of copper (I) iodide were added. The solution was combined with 320 μ L H u nig base and then 18mg (0.27mmol, 3eq) of ethynylcyclopropane. The reaction mixture was passed through a column with DCM/MeOH/NH₃4/1/0.1 mixture was filtered through silica gel, and 6mL of RP-gel was added. After removal of volatiles, column chromatography purification was completed by RP-phase (changing from 95% water (+ 0.2% formic acid) and 5% acetonitrile (+ 0.2% formic acid) to 50% water and 50% acetonitrile in 20 min). The corresponding product components were combined and lyophilized to remove the solvent. 32mg (0.065mmol, 71%) of compound 137 are obtained.

Example 138-139 was prepared similarly, however in example 138 the reaction was completed under propyne gas in a nitrogen cylinder at 40 ℃.

Example 140

(±)-4-[4-((1R^*，2S^*) -2-carbamoyl-cyclopentylamino) -5-cyclopropyl-pyrimidin-2-ylamino]-N- (1-methyl-piperidin-4-yl) -benzamide (FIG. E)

100mg (0.15mmol) of 104 was suspended in 1.4mL dioxane and 13mg (0.15mmol, 1eq) cyclopropylboronic acid was added. The solution was degassed in vacuo and 3.5mg (0.004mmol, 3 mol%) of dichloro [1, 1' -bis (diphenylphosphino) -ferrocene]Palladium (II) -dichloromethane adduct (PdCl)₂dppf DCM) and 2mL of sodium carbonate solution (2M in water) were added under argon. The biphasic mixture was heated at 130 ℃ for 5 minutes (CEM microwave, 100W). The organic phase was separated, diluted with methanol and combined with 6mL of RP-gel. After removal of volatiles, purification was accomplished by reverse phase (changing from 97% water (+ 0.2% formic acid) and 3% acetonitrile (+ 0.2% formic acid) to 70% water and 30% acetonitrile over 12 minutes) by column chromatography. The corresponding product components were combined and lyophilized to remove the solvent. 2mg (0.003mmol, 2%) of compound 140 are obtained.

Example 141

(±)-(1S^*，2R^*)-2-[2-(4-[1.4]Diazepan-1-yl-3-fluoro-phenylamino) -5-trifluoromethyl-pyrimidin-4-ylamino]-cyclopentylAlkanecarboxylic acid isopropylamide (FIG. B)

23mg (0.066mmol) of B-2a are dissolved in 100. mu.L of NMP, 17mg (0.079mmol, 1.2eq) of 3-fluoro-4- (4-methyl- [1.4] diazepan-1-yl) -aniline are added, and finally 46. mu.L of HCl (0.18mmol, 2.8eq, 4M in dioxane) are added. The reaction mixture was heated at 90 ℃ for 12 h, combined with 6 mLRP-gel and the volatiles were removed in vacuo. Chromatographic purification was done by reverse phase (changing from 95% water (+ 0.2% formic acid) and 5% acetonitrile (+ 0.2% formic acid) to 55% water and 45% acetonitrile over 25 minutes). The corresponding product components were combined and lyophilized to remove the solvent. 3mg (0.005mmol, 8%) of compound 141 are obtained.

Example 142 was prepared similarly to example 144.

Example 145

(±)-(1R^*，2R^*) -2- {2- [4- (4-methyl-piperazine-1-carbonyl) -phenylamino]-5-trifluoromethyl-pyrimidin-4-ylamino } -cyclopentanecarboxamide (scheme C)

100mg (0.25mmol) of C-2a are dissolved in 1mL of 1-butanol and this solution is combined with 35mg (0.275mmol, 1.1eq) of racemic trans-2-aminocyclopentanecarboxamide and 60. mu.L (0.35mmol, 1.4eq) of Hunig base. The mixture was stirred at 110 deg.C (100W, microwave CEM) for 30 minutes until the conversion was complete. About 20mL of methanol was added to the reaction mixture, which was combined with the RP-gel (about 8mL), and all volatiles were removed in vacuo. The mixture was purified by RP column (changing from 95% water (+ 0.2% formic acid) and 5% acetonitrile (+ 0.2% formic acid) to 55% water and 45% acetonitrile in 20 minutes). The corresponding product fractions were combined with concentrated hydrochloric acid and lyophilized to remove the solvent. 77mg (0.146mmol, 58%) of compound 145 are obtained as a colorless solid.

Example 146-147 was prepared analogously, while example 148 was prepared analogously to example 129 (nucleophilic substitution of the. beta. -amino acid starting from C-2a and finally amide formation by binding to ammonia).

Examples 1 to 148

These examples describe the biological activity of the compounds of the invention and do not limit the invention to these examples.

The inhibition of proliferation by the compounds of the invention was first mediated by errors in chromosome segregation, as demonstrated by DNA staining followed by FACS or Cellomics Array Scan analysis. Due to the accumulation of erroneous separations, a large number of polyploids appear which can ultimately lead to inhibition of proliferation or even apoptosis. Based on their biological properties, the compounds of the general formula (I) according to the invention, their isomers and their physiologically usable salts are suitable for the treatment of diseases which are characterized by excessive or abnormal cell proliferation.

Examples Aurora-B kinase assay

A radioenzyme inhibition assay was developed using a baculovirus-expressed recombinant human Aurora B wild-type protein with a histidine (6) epitope (His-) at the N-terminal position, which was obtained from infected insect cells (SF21), and purified.

Expression and purification

For this purpose, 300X10 in SF-900II insect cell culture medium (Invitrogen)⁶SF21 cells are incubated, for example, with an appropriate amount of baculovirus solution at 27 ℃ for 1 hour (Fernbach shaker bottle, 50 rpm). Then 250ml SF-900II medium was added and shaken for 3 days (100rpm, 27 ℃). Three hours before harvest, okadaic acid (C) was added₄₄H₆₈O₁₃Calbiochem #495604) (final concentration 0.1. mu.M) to stabilize the phosphorylation site on recombinant Aurora B. The cells were centrifuged (1000rpm, 5 min, 4 ℃) to obtain pellet, the supernatant was removed and the pellet was frozen in liquid nitrogen. The pellet was thawed (37 ℃, 5 min) and resuspended in lysis buffer. 40mL lysis buffer (25mM Tris/Cl, 10mM MgCl)₂300mM NaCl, 20mM imidazole, pH8.0, 0.07% 2-mercaptoethanol and protease inhibitors all from Roche Diagnostics) was used in a 200mL volume of the initial culture. After two rapid freeze/thaw cycles (liquid nitrogen at 37 ℃), the lysates were stored on ice for 30 min and then incubated with washed Ni-NTA Beads (Ni-NTA Superflow Beads, 4mL/200mL of starting culture) (2 h, 4 ℃) in an Econo-Pac column (Biorad #732- "1010). Elution buffer (25mM Tris/Cl, 10mM MgCl) in 10 column volumes in each case₂1000mM NaCl, 20mM imidazole, pH8.0, 0.07% 2-mercaptoethanol and protease inhibitors were all from Roche Diagnostics) 5 washes, followed by elution with 8ml (per 200ml of starting culture) of elution buffer (25mM Tris/Cl pH8.0, 300mM NaCl, 10mM MgCl2, 0.03% Brij-35, 10% glycerol, 0.07% 2-mercaptoethanol, 400mM imidazole). Desalting the combined eluted fractions with Sephadex G25 column andtransfer to a freezing buffer (50mM tris/Cl pH8.0, 150mM NaCl, 0.1mM EDTA, 0.03% Brij-35, 10% glycerol, 1mM DTT).

Kinase assay

The samples were placed in polypropylene dishes (96 wells, Greiner # 655201) covering a concentration range of 10. mu.M to 0.0001. mu.M. The final concentration of DMSO in the assay was 5%. mu.L of the protein mixture (50mM tris/Cl pH7.5, 25mM MgCl)₂25mM NaCl, 167. mu.M ATP, 200ng His-Aurora B in the freeze buffer) was added to 10. mu.l of 25% DMSO sample, which was incubated for 15 minutes at room temperature. 10 μ L of peptide mixture (100mM tris/Cl pH7.5, 50mM MgCl) was then added₂，50mM NaCl，5μM NaF，5M DTT，1μ Ciγ-P33-ATP[Amersham]50 μ M substrate peptide [ biotin-EPLERRLSLVPDS or multimers thereof, or biotin-EPLERRLSLVPKM or multimers thereof, or biotin-LRRWSLGLRRWSLGLRRWSLGLRRWSLG]). The reaction was incubated for 75 minutes (room temperature), stopped by the addition of 180. mu.L of 6.4% trichloroacetic acid, and incubated on ice for 20 minutes. The multi-mesh filter plate (Millipore, MAIP NOB10) was first equilibrated with 100. mu.L of 70% ethanol and then with 180. mu.L of trichloroacetic acid, and the liquid was removed with a suitable suction filtration device. The kinase reaction was then stopped. After washing 5 times with 180. mu.L of 1% trichloroacetic acid in each case, the lower half of the dish was dried (10-20 minutes at 55 ℃) and 25. mu.L of scintillation cocktail (Microscint, Packard #6013611) was added. Gamma-phosphate was dosed using a Wallac1450 Microbeta Liquid science Counter. Samples without sample or substrate peptide were used as controls. IC Using Graph Pad Prism software₅₀The value is obtained.

The antiproliferative activity of the compounds of the invention is determined in cultured human tumor cell proliferation assays and/or in cell cycle assays, such as NCI-H460 tumor cells. The compounds show good or very good activity in both test methods, i.e. an EC50 value of less than 5. mu. mol/L, usually less than 1. mu. mol/L, for example in the NCI-H460 proliferation test.

Assays for inhibition of proliferation of cultured human tumor cells

To determine the proliferation of cultured human tumor cells, cells of the lung tumor cell line NCI-H460 (obtained from American Type Culture Collection (ATCC)) were cultured in RPMI 1640 medium (Gibco) and 10% fetal bovine serum (Gibco) and harvested in the logarithmic growth phase. NCI-H460 cells were then plated at a density of 1000 cells per well in RPMI 1640 medium in 96-well flat bottom plates (Falcon) in an incubator (at 37C and 5% CO)₂) Cultured overnight in the medium. The active substance was added to the cells at various concentrations (dissolved in DMSO; DMSO final concentration: 0.1%). After 72 hours of incubation, 20. mu.l of AlamarBlue reagent (AccuMed International) was added to each well and the cells were further incubated for 5-7 hours. After incubation, the color change of AlamarBlue reagent was measured in a Wallac Microbeta fluorescence spectrophotometer. EC was calculated using the standard Levenburg Marquard algorithm (GraphPadPrizm)₅₀The value is obtained. Cell cycle analysis is carried out, for example, by FACS analysis (fluorescence activated Cell Sorter) or by the cellular Array Scan (Cell cycle analysis).

FACS analysis

Propidium Iodide (PI) binds stoichiometrically to double-stranded DNA and is therefore suitable for determining the proportion of cells in the G1, S and G2/M phases of the cell cycle, based on the cellular DNA content. Cells in G0 and G1 have diploid DNA content (2N), while cells in G2 or the mitotic phase have 4N DNA content.

For PI staining, for example, 0.4 million 1.75X10⁶NCI-H460 cells were seeded at 75cm²On cell culture flasks, after 24 hours either 0.1% DMSO was added as a control or substrate was added at various concentrations (in 0.1% DMSO). Cells were incubated with substrate or with DMSO for 42 hours. The cells were then detached with trypsin and centrifuged. The cell pellet was washed with buffered saline (PBS) and then the cells were fixed at 80% concentration for at least 2 hours at-20 ℃. After additional PBS washing, the cells were dialyzed against Triton X-100 (Sigma; 0.25% in PBS) on ice for 5 minutes and then incubated with a 9: 1 solution of propidium iodide (Sigma; 10. mu.g/ml) and RNase (Serva; 1mg/mLl) in the dark for at least 20 minutes.

DNA determination was done in a Becton Dickinson FACS Analyzer with an argon laser (500mW, emission wavelength 488 nm); the data obtained were evaluated by DNA Cell Quest program (BD).

Cellomics array scanning

NCI-H460 cells were seeded at a density of 2000 cells per well in RPMI 1640 medium (Gibco) with 10% fetal bovine serum (Gibco) in 96-well plates (Falcon) in an incubator (at 37 ℃ and 5% CO)₂) Cultured overnight in the medium. The active substance was added to the cells at various concentrations (dissolved in DMSO; DMSO final concentration: 0.1%). After 42 hours of incubation, the medium was filtered with suction, and the cells were fixed with 4% formaldehyde solution and Triton X-100 (1: 200 in PBS) for 10 minutes at room temperature while dialyzing, and then washed twice with 0.3% BSA solution (Calbiochem). Then, 50. mu.L/well of 4', 6-diamidino-2-phenylindole (DAPI; Molecular Probes) was added to the DNA at a final concentration of 300nM and stained in the dark at room temperature for 1 hour. The product was then carefully washed twice with PBS, plates were stuck with a black sticky membrane, analyzed and displayed in a Cellomics ArrayScan using the CellCycle Bioapplication program, and evaluated with Spotfire.

The agents of the invention are Aurora kinase inhibitors. Based on their biological properties, the compounds of the general formula (I) according to the invention, their isomers and their physiologically usable salts are suitable for the treatment of diseases which are characterized by excessive or abnormal cell proliferation.

Such diseases include, for example: viral infections (e.g., HIV and kaposi's sarcoma); inflammatory and autoimmune diseases (e.g., colitis, arthritis, alzheimer's disease, glomerulonephritis and wound healing); bacterial, fungal and/or parasitic infections; leukemias, lymphomas, and solid tumors (e.g., carcinomas and sarcomas), skin diseases (e.g., psoriasis); proliferative diseases characterized by an increased number of cells (e.g., fibroblasts, hepatocytes, bone and bone marrow cells, cartilage or smooth muscle cells, or epithelial cells (e.g., endometrial hyperplasia)); bone diseases and cardiovascular diseases (e.g. restenosis and hypertrophy).

For example, the following cancers may be treated with the compounds of the present invention, but are not limited to: brain tumors such as, for example, acoustic schwannoma (neuroma), astrocytoma such as glioma, fibroastrocytoma, plasmogenic astrocytoma, mastocytoma, glioblastoma multiforme and glioblastoma, brain lymphoma, brain metastases, pituitary tumors such as prolactin tumors, HGH (human (pituitary) growth hormone) producing tumors and ACTH producing tumors (adrenocorticotropic hormone), craniopharyngioma, medulloblastoma, meningioma and oligodendroglioma; neuromas (neoplasms) such as, for example, autonomic nervous system tumors such as neuroblastoma, ganglionic tumor (pheochromocytoma, paraganglioma) and carotid aneurysm, peripheral nervous system tumors such as amputation neuroma, neurofibroma, Schwannoma and malignant Schwannoma and central nervous system tumors such as brain and myeloma; bowel cancers such as, for example, rectal, colon, anal, small intestine, and duodenal cancers; eyelid tumors such as basal cell carcinoma or basal cell carcinoma; pancreatic cancer; bladder cancer; lung cancer (bronchial cancer) such as, for example, small cell bronchial cancer (oat [ cell ] carcinoma) and non-small cell bronchial cancer such as plate epithelial cancer, adenocarcinoma, and large cell bronchial cancer; breast cancers such as, for example, breast cancers such as invasive ductal carcinoma, mucinous carcinoma, lobular invasive carcinoma, tubular carcinoma, cystic adenoid carcinoma, and papillary carcinoma; non-hodgkin's lymphomas (NHLs) such as, for example, Burkitt's lymphoma, low malignancy non-hodgkin's lymphoma (NHL), and mucosis fungoides; uterine or endometrial cancer or uterine body cancer; CUP syndrome; ovarian cancer such as mucinous, endometrial or serum cancers; gallbladder cancer; bile duct cancers such as, for example, Klatskin tumor; testicular cancers such as, for example, seminoma and non-seminoma; lymphomas (lymphosarcomas) such as, for example, malignant lymphomas, Hodgkin's disease, non-Hodgkin's lymphomas (NHL) such as chronic lymphocytic leukemia, leukemia reticuloendotheliosis, immunocytomas, plasmacytomas (multiple myeloma), immunoblastomas, Burkitt's lymphoma, T-zone mycosis fungoides, large cell anaplastic lymphoblastomas, and lymphoblastomas; laryngeal cancers such as, for example, vocal cord tumors, supraglottic tumors, glottic tumors, and subglottic laryngeal tumors; bone cancers such as, for example, osteochondroma, chondroma, chondroblastoma, chondrmucoid fibroma, osteoma, osteoid osteoma, osteoblastoma, eosinophilic granuloma, giant cell tumor, chondrosarcoma, osteosarcoma, ewing's sarcoma, reticulosarcoma, plasmacytoma, giant cell tumor, fibrous dysplasia, juvenile bone cyst, and aneurysm bone cyst; head and neck cancers such as, for example, lip cancer, tongue cancer, cancer of the floor of the mouth, cancer of the oral cavity, cancer of the gingiva, cancer of the palate, salivary glands, cancer of the pharynx, cancer of the nasal cavity, cancer of the paranasal sinuses, larynx, and middle ear; liver cancers such as, for example, hepatocellular carcinoma or hepatocellular carcinoma (HCC); leukemias, such as, for example, acute leukemias, e.g., acute lymphoblastic/lymphoblastic leukemia (ALL), Acute Myeloid Leukemia (AML); chronic leukemias such as Chronic Lymphocytic Leukemia (CLL), Chronic Myelogenous Leukemia (CML); gastric or stomach cancers such as, for example, papillary, ductal and mucinous adenocarcinoma, signet ring cell carcinoma, adenosquamous carcinoma, small cell carcinoma and undifferentiated carcinoma; melanomas such as, for example, epilentine, nodular, lentigo maligna, and acromelasma melanose; kidney cancers such as, for example, renal cell carcinoma or glavier's or Grawitz's; esophageal cancer or esophageal cancer; penile cancer; prostate cancer; laryngeal or pharyngeal cancer such as, for example, nasopharyngeal cancer, oropharyngeal cancer, and laryngopharyngeal cancer; retinoblastoma such as, for example, vaginal cancer; plate epithelial carcinoma, adenocarcinoma, carcinoma in situ, malignant melanoma, and sarcoma; thyroid cancers such as, for example, papillary, follicular and medullary thyroid cancers and anaplastic cancers; spinolioma, epidomoid cancer, and skin plate epitolial cancer; thymoma, cancer of the urethra, and cancer of the vulva.

The novel compounds of the invention are useful for the prophylaxis, short-term or long-term treatment of the abovementioned diseases, optionally in combination with radiotherapy or other "state of the art" compounds, such as, for example, cytostatic or cytotoxic substances, cytostatic agents, antiangiogenic substances, steroids or antibodies.

According to the invention, the compounds of the general formula (1) can be used on their own or in combination with other active substances, optionally also with other physiologically active substances.

Chemotherapeutic agents that can be used in combination with the compounds of the invention include, but are not limited to, hormones, hormone analogs, and anti-hormones (e.g., tamoxifen, toremifene, raloxifene, fulvestrant, megestrol acetate, flutamide, nilutamide, bicalutamide, aminoglutethimide, cyproterone acetate, finasteride, buserelin acetate, fludrocortisone, flumestosterone, medroxyprogesterone, octreotide), aromatase inhibitors (e.g., anastrozole, letrozole, liazole, vorozolozole, exemestane, atacetam), LHRH agonists and antagonists (e.g., capromorelin acetate, luprolide), growth factor inhibitors (growth factors such as, for example, "platelet-derived growth factor" and "hepatocyte growth factor", inhibitors such as, for example, "growth factor" antibodies, "growth factor receptor" antibodies, and tyrosine kinase inhibitors, such as, for example, gefitinib, imatinib, lapatinib, and trastuzumab); antimetabolites (e.g., folic acid antagonists such as methotrexate, raltitrexed, pyrimidine analogs such as 5-fluorouracil, capecitabine and gemcitabine, purines, and adenosine analogs such as mercaptopurine, thioguanine, cladribine and pentostatin, cytarabine, fludarabine); antibiotics with antitumor effect (e.g., anthracyclines such as doxorubicin, daunorubicin, epirubicin, and demethoxydaunorubicin, mitomycin C, bleomycin, actinomycin D, plicamycin, streptozocin); platinum derivatives (e.g., cisplatin, oxaliplatin, carboplatin); alkylating agents (e.g., estramustine, meclorethamine, melphalan, chlorambucil, busulfan, dacarbazine, cyclophosphamide, ifosfamide, temozolomide, nitrosoureas such as, for example, carmustine and lomustine, thiotepa); anti- (mitotic) splitting agents (e.g. vinca alkaloids such as e.g. vinblastine, vindesine, vinorelbine and vincristine; and taxanes such as paclitaxel, docetaxel); topoisomerase inhibitors (e.g. etoposide such as e.g. etoposide and valcepide, teniposide, amsacrine, topotecan, irinotecan, mitoxantrone) and various chemotherapeutic agents such as amifostine, anagrelide, clodronate, filgrastim, interferon alpha, calcium folinate, merozoite, procarbazine, levamisole, mesna, mitotane, disodium pamidronate and porfilline sodium.

Suitable formulations include, for example, tablets, capsules, suppositories, solutions-especially for injection (subcutaneous, intravenous, intramuscular) and infusion solutions-and elixirs, emulsions or dispersible powders. The amount of pharmaceutically active compound should be in the range of 0.1 to 90% by weight, preferably 0.5 to 50% by weight, of the total composition, i.e. in an amount sufficient to achieve the following specific dosage ranges. If necessary, specific doses may be administered several times a day.

Suitable tablets are prepared, for example, by mixing the active substance with known excipients, for example inert diluents, such as calcium carbonate, calcium phosphate or lactose, disintegrants, such as corn starch or alginic acid, binders, such as starch or gelatin, lubricants, such as magnesium stearate or talc, and/or slow-release agents, such as carboxymethylcellulose, cellulose acetate phthalate or polyvinyl acetate. The tablet may also comprise several layers.

The coated tablets can accordingly be prepared by film-coating tablets, analogously to tablets with the usual tablet coating substances, for example by colladone or shellac, gum arabic, talc, titanium dioxide or sugar. The core may also be composed of a number of layers to achieve sustained release or to prevent incompatibility. Similarly, the tablet coating may be composed of a number of layers to achieve sustained release, possibly using the excipients mentioned above for the tablets.

A syrup or elixir containing the active substance or a composition thereof according to the invention may also contain a sweetening agent such as saccharin, cyclamate, glycerol or sugar and a flavoring agent, for example a flavoring agent such as vanillin or citrus extract. They may also contain suspension adjuvants or thickeners such as sodium carboxymethylcellulose, wetting agents such as, for example, condensation products of fatty alcohols with ethylene oxide, or preservatives such as p-hydroxybenzoic acid.

Solutions for injection and infusion are prepared in a conventional manner, for example by adding isotonic agents, preservatives such as p-hydroxybenzoic acid or stabilizers such as alkali metal salts of ethylenediamine tetraacetic acid, optionally with emulsifiers and/or dispersants, while if water is used as diluent, for example, organic solvents may optionally be used as solvents or cosolvents, and transferred into injection bottles or ampoules or infusion bottles.

Capsules containing one or more active substances or compositions of active substances can be prepared, for example, by mixing the active substances with inert carriers, such as lactose or sorbitol, and filling them in gelatin capsules.

Suitable suppositories may be prepared, for example, by mixing with carriers provided for this purpose, such as neutral fats or polyethylene glycols or derivatives thereof.

Useful excipients include, for example, water, pharmaceutically acceptable organic solvents such as paraffin oils (e.g., petroleum fractions), vegetable oils (e.g., peanut or sesame oil), mono-or polyfunctional alcohols (e.g., ethanol or glycerol), carriers such as, for example, natural mineral powders (e.g., kaolin, clay, talc, chalk), synthetic mineral powders (e.g., highly dispersed silicic acid and silicates), sugar (e.g., sucrose, lactose and glucose) emulsifiers (e.g., lignin, sucrose, methyl cellulose, starch and polyvinylpyrrolidone), and lubricants (e.g., magnesium stearate, talc, stearic acid and sodium lauryl sulfate).

These formulations are used by conventional means, preferably by the oral or transdermal route, most preferably by the oral route. For oral administration, of course, the tablet may contain, in addition to the above-mentioned carrier, additives such as sodium citrate, calcium carbonate and dicalcium phosphate and various additives such as starch, preferably potato starch, gelatin and the like. Additionally, lubricants such as magnesium stearate, sodium lauryl sulfate and talc may be used simultaneously for tableting. For aqueous suspensions, the active substance may be combined with various adjuvants or colorants in addition to the excipients.

For parenteral use, solutions of the active substance in a suitable liquid carrier may be used.

The dosage to be used intravenously is 1-1000mg per hour, preferably 5-500 mg per hour.

However, it may sometimes be necessary to employ specific amounts depending on the body weight, route of use, individual response to the drug, nature of the formulation, and time or interval of use of the drug. Thus, in some cases it may be sufficient to use a lower minimum dose than given above, while in other cases the upper limit may have to be exceeded. When administered in large amounts, it may be desirable to divide them into many smaller doses throughout the day.

The following formulation examples illustrate the invention without limiting its scope:

examples of pharmaceutical preparations

A) Tablet formulation Each sheet is

Active substance 100mg

Lactose 140mg

Corn starch 240mg

Polyvinylpyrrolidone 15mg

Magnesium stearate 5mg

500mg

Finely ground active, lactose and some corn starch are mixed together. The mixture was sieved, then wetted with an aqueous solution of polyvinylpyrrolidone, kneaded to form wet granules, and dried. The granules, residual corn starch and magnesium stearate are sieved and mixed together. The mixture is compressed to produce tablets of suitable shape and size.

B) Tablet formulation Each sheet is

80mg of active substance

Lactose 55mg

Corn starch 190mg

Microcrystalline cellulose 35mg

Polyvinylpyrrolidone 15mg

Sodium carboxymethyl starch 23mg

Magnesium stearate 2mg

400mg

Finely ground active material, some corn starch, lactose, microcrystalline cellulose and polyvinylpyrrolidone are mixed together, the mixture is sieved and treated with residual corn starch and water to form dried and sieved granules. Sodium carboxymethyl starch and magnesium stearate are added, mixed and the mixture is compressed to form tablets of appropriate size.

C) Ampoule solution

Active substance 50mg

50mg of sodium chloride

5ml of water for injection

The active substance is dissolved in water at its own pH or optionally at pH5.5 to 6.5 and made isotonic by addition of sodium chloride. Filtering the obtained solution, removing heat source, transferring the filtrate into ampoule under aseptic condition, sterilizing, and melting and sealing. Ampoules contain 5mg, 25mg and 50mg of active substance.

Reference to the literature

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Andrews，P.D.Knatko，E.Moore，W.J.and Swedlow，J.R.(2003).Mitoticmechanics：the auroras come into view.Curr.Opin.Cell Biol.15，672-683.

Bayliss，R.Sardon，T.Vernos，I.and Conti，E.(2003).Structural basis ofAurora-A activation by TPX2 at the mitotic spindle.Mol.Cell 12，851-862.

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Claims (15)

1. A compound of the general formula (1)

Wherein

R¹Is represented by R⁵And optionally substituted by one or more R⁴A substituted group selected from C_3-10-cycloalkyl and 3-8 membered heterocycloalkyl;

R²is represented by optionally one orPlural R⁴A substituted group selected from C_1-6Alkyl radical, C_3-10-cycloalkyl, 3-8 membered heterocycloalkyl, C_6-15-aryl and 5-12 membered heteroaryl;

R³represents hydrogen, halogen, -CN, -NO₂、C_1-4Alkyl radical, C_1-4-haloalkyl group, C_3-10-cycloalkyl, C_4-16-cycloalkylalkyl and C_7-16-a radical of arylalkyl;

R⁴represents a group selected from R^a、R^bAnd by one or more R, which may be the same or different^cAnd/or R^bSubstituted R^aA group of (a);

R⁵represents a group selected from-C (O) R^c、-C(O)NR^cR^c、-S(O)₂R^c、-N(R^f)S(O)₂R^c、-N(R^f)C(O)R^c、-N(R^f)C(O)OR^cand-N (R)^f)C(O)NR^cR^cA group of (a);

each R^aIndependently of one another, from C_1-6Alkyl radical, C_3-10-cycloalkyl, C_4-16-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl;

each R^bAre suitable radicals and are in each case independently of one another selected from ═ O, -OR^c、C_1-3Haloalkoxy, -OCF₃、＝S、-SR^c、＝NR^c、＝NOR^c、-NR^cR^cHalogen, -CF₃、-CN、-NC、-OCN、-SCN、-NO₂、-S(O)R^c、-S(O)₂R^c、-S(O)₂OR^c、-S(O)NR^cR^c、-S(O)₂NR^cR^c、-OS(O)R^c、-OS(O)₂R^c、-OS(O)₂OR^c、-OS(O)₂NR^cR^c、-C(O)R^c、-C(O)OR^c、-C(O)NR^cR^c、-CN(R^f)NR^cR^c、-CN(OH)R^c、-CN(OH)NR^cR^c、-OC(O)R^c、-OC(O)OR^c、-OC(O)NR^cR^c、-OCN(R^f)NR^cR^c、-N(R^f)C(O)R^c、-N(R^f)C(S)R^c、-N(R^f)S(O)₂R^c、-N(R^f)C(O)OR^c、-N(R^f)C(O)NR^cR^c、-[N(R^f)C(O)]₂R^c、-N[C(O)]₂R^c、-N[C(O)]₂OR^c、-[N(R^f)C(O)]₂OR^cand-N (R)^f)CN(R^f)NR^cR^c；

Each R^cIndependently of one another, hydrogen or optionally substituted by one or more identical or different R^dAnd/or R^eSubstituted groups selected from: c_1-6Alkyl radical, C_3-10-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl,

each R^dIndependently of one another, hydrogen or optionally substituted by one or more identical or different R^eAnd/or R^fSubstituted groups selected from: c_1-6Alkyl radical, C_3-8-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl;

each R^eAre suitable radicals and are each, independently of one another, selected from ═ O, -OR^f、C_1-3Haloalkoxy, -OCF₃、＝S、-SR^f、＝NR^f、＝NOR^f、-NR^fR^fHalogen, -CF₃、-CN、-NC、-OCN、-SCN、-NO₂、-S(O)R^f、-S(O)₂R^f、-S(O)₂OR^f、-S(O)NR^fR^f、-S(O)₂NR^fR^f、-OS(O)R^f、-OS(O)₂R^f、-OS(O)₂OR^f、-OS(O)₂NR^fR^f、-C(O)R^f、-C(O)OR^f、-C(O)NR^fR^f、-CN(R^g)NR^fR^f、-CN(OH)R^f、-C(NOH)NR^fR^f、-OC(O)R^f、-OC(O)OR^f、-OC(O)NR^fR^f、-OCN(R^g)NR^fR^f、-N(R^g)C(O)R^f、-N(R^g)C(S)R^f、-N(R^g)S(O)₂R^f、-N(R^d)C(O)OR^f、-N(R^g)C(O)NR^fR^fAnd N (R)^g)CN(R^f)NR^fR^f；

Each R^fIndependently of one another, hydrogen or optionally substituted by one or more identical or different R^gSubstituted groups selected from: c_1-6Alkyl radical, C_3-8-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl alkyl, 5-12 membered heteroaryl, and 6-18 membered heteroarylalkyl;

each R^gIndependently of one another are hydrogen, C_1-6Alkyl radical, C_3-8-cycloalkyl, C_4-11-cycloalkylalkyl, C_6-10Aryl radical, C_7-16Arylalkyl, 2-6 membered heteroalkyl, 3-8 membered heterocycloalkyl, 4-14 membered heterocycloalkyl, 5-12 membered heteroaryl and 6-18 membered heteroarylalkyl, optionally in the form of tautomers, racemates, enantiomers, diastereomers and mixtures thereof, and optionally in the form of pharmaceutically acceptable acid addition salts thereof.

2. A compound according to claim 1, wherein R³Represents a group selected from halogen and C_1-4A haloalkyl group.

3. A compound according to claim 2, wherein R³represents-CF₃。

4. A compound according to any one of claims 1 to 3, wherein R²Is represented by C_6-10Aryl or 5-12 membered heteroaryl, optionally substituted by one or more R⁴And (4) substitution.

5. A compound according to claim 4, wherein R²Represents phenyl, optionally substituted by one or more R⁴And (4) substitution.

6. A compound of the general formula (1A),

wherein

n is equal to 0 or 1 and,

m is equal to 1-5 and

y is equal to 0 to 6 and the remaining groups are as defined above.

7. A compound according to claim 6, wherein R³Represents a group selected from halogen and C_1-4A haloalkyl group.

8. A compound according to claim 7, wherein R³Denotes CF₃。

9. A compound according to any one of claims 6 to 8, wherein R²Is represented by C_6-10Aryl or 5-12 membered heteroaryl, optionally substituted by one or more R⁴And (4) substitution.

10. A compound according to any one of claims 6 to 9, wherein R²Represents phenyl, optionally substituted by one or more R⁴And (4) substitution.

11. A compound according to any one of claims 1 to 10, or a pharmaceutically active salt thereof, for use in a pharmaceutical composition.

12. Use of a compound according to any one of claims 1 to 10, or a pharmaceutically active salt thereof, for the preparation of a pharmaceutical composition having antiproliferative activity.

13. Pharmaceutical preparation comprising as active substance one or more compounds of the general formula (1) or (1A) according to any one of claims 1 to 10 or a physiologically acceptable salt thereof, optionally in combination with customary excipients and/or carriers.

14. Use of a compound of general formula (1) or (1A) according to any one of claims 1 to 10 for the preparation of a pharmaceutical composition for the treatment and/or prevention of cancer, infections, inflammations and autoimmune diseases.

15. Pharmaceutical preparation comprising a compound of general formula (1) or (1A) according to claims 1 to 10 and at least one other cytostatic or cytotoxic substance different from general formula (1) or (1A), optionally in the form of tautomers, racemates, enantiomers, diastereomers and mixtures thereof, and optionally in the form of pharmaceutically acceptable acid addition salts thereof.