CDK-inhibitory pyrimidines, their production and use as pharmaceutical agents

This application is a divisional of U.S. patent application Ser. No. 10/156,759, filed Nov. 7, 2002.

This invention relates to pyrimidine derivatives, their production as well as their use as medications for treating various diseases.

The CDKs (cyclin-dependent kinase) is an enzyme family that plays an important role in the regulation of the cell cycle and thus is an especially advantageous target for the development of small inhibitory molecules. Selective inhibitors of the CDKs can be used for treating cancer or other diseases that cause disruptions of cell proliferation.

Pyrimidines and analogs are already described as active ingredients, such as, for example, the 2-anilino-pyrimidines as fungicides (DE 4029650) or substituted pyrimidine derivatives for treating neurological or neurodegenerative diseases (WO 99/19305). As CDK inhibitors, the most varied pyrimidine derivatives are described, for example bis(anilino)-pyrimidine derivatives (WO 00/12486), 2-amino-4-substituted pyrimidines (WO 01/14375), purines (WO 99/02162), 5-cyano-pyrimidines (WO 02/04429), anilinopyrimidines (WO 00/12486) and 2-hydroxy-3-N,N-dimethylaminopropoxy-pyrimidines (WO 00/39101).

The object of this invention is to provide compounds that have better properties than the inhibitors that are already known. The substances that are described here are more effective, since they already inhibit in the nanomolar range and can be distinguished from other already known CDK inhibitors such as, e.g., olomoucine and roscovitine.

It has now been found that compounds of general formula I

in which

R 1 stands for hydrogen, halogen, C 1 -C 6 -alkyl, nitro, or for the group —COR 5 , —OCF 3 , —(CH 2 ) n R 5 , —S—CF 3 or —SO 2 CF 3 , R 2 stands for C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkinyl, or C 3 -C 10 -cycloalkyl or for C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkinyl, or C 3 -C 10 -cycloalkyl that is substituted in one or more places in the same way or differently with hydroxy, halogen, C 1 -C 6 -alkoxy, C 1 -C 6 -alkylthio, amino, cyano, C 1 -C 6 -alkyl, —NH—(CH 2 ) n —C 3 -C 10 -cycloalkyl, C 3 -C 10 -cycloalkyl, C 1 -C 6 -hydroxyalkyl, C 2 -C 6 -alkenyl, C 2 -C 6 -alkinyl, C 1 -C 6 -alkoxy-C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy-C 1 -C 6 -alkoxy-C 1 -C 6 -alkyl, —NHC 1 -C 6 -alkyl, —N(C 1 -C 6 -alkyl) 2 , —SO(C 1 -C 6 -alkyl), —SO 2 (C 1 -C 6 -alkyl), C 1 -C 6 -alkanoyl, —CONR 3 R 4 , —COR 5 , C 1 -C 6 -alkylOAc, carboxy, aryl, heteroaryl, —(CH 2 ) n -aryl, —(CH 2 ) n -heteroaryl, phenyl-(CH 2 ) n —R 5 , —(CH 2 ) n PO 3 (R 5 ) 2 or with the group —R 6 or —NR 3 R 4 , and the phenyl, C 3 -C 10 -cycloalkyl, aryl, heteroaryl, —(CH 2 ) n -aryl and —(CH 2 ) n -heteroaryl itself optionally can be substituted in one or more places in the same way or differently with halogen, hydroxy, C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy, heteroaryl, benzoxy or with the group —CF 3 or —OCF 3 , and the ring of the C 3 -C 10 -cycloalkyl and the C 1 -C 10 -alkyl optionally can be interrupted by one or more nitrogen, oxygen and/or sulfur atoms and/or can be interrupted by one or more ═C═O groups in the ring and/or optionally one or more possible double bonds can be contained in the ring, or R 2 stands for the group

X stands for oxygen or for the group —NH—, —N(C 1 -C 3 -alkyl) or for —OC 3 -C 10 -cycloalkyl, which can be substituted in one or more places in the same way or differently with a heteroaromatic compound,

or

X and R 2 together form a C 3 -C 10 -cycloalkyl ring, which optionally can contain one or more heteroatoms and optionally can be substituted in one or more places with hydroxy, C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy or halogen,

A and B, in each case independently of one another, stand for hydrogen, hydroxy, C 1 -C 3 -alkyl, C 1 -C 6 -alkoxy or for the group —SR 7 , —S(O)R 7 , —SO 2 R 7 , —NHSO 2 R 7 , —CH(OH)R 7 , —CR 7 (OH)—R 7 ,

C 1 -C 6 -alkylP(O)OR 3 OR 4 or —COR 7 , or for

whereby the phenyl itself can be substituted in one or more places in the same way or differently with halogen, C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy or with the group —SO 2 NR 3 R 4 , or for the group —(CH 2 ) n NR 3 R 4 , —CNHNH 2 or —NR 3 R 4 , or

R 3 and R 4 together form a C 3 -C 10 -cycloalkyl ring that optionally can be interrupted by one or more nitrogen, oxygen and/or sulfur atoms and/or can be interrupted by one or more ═C═O groups in the ring and/or optionally one or more possible double bonds can be contained in the ring,

R 5 stands for hydroxy, phenyl, C 1 -C 6 -alkyl, C 3 -C 6 -cycloalkyl, benzoxy, C 1 -C 6 -alkylthio or C 1 -C 6 -alkoxy,

R 6 stands for a heteroaryl or C 3 -C 10 -cycloalkyl ring, whereby the ring has the above-indicated meaning,

R 7 stands for halogen, hydroxy, phenyl, C 1 -C 6 -alkyl, C 2 -C 6 -alkenyl, C 2 -C 6 -alkinyl, C 3 -C 10 -cycloalkyl, with the above-indicated meaning, or for the group —NR 3 R 4 , or for C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkinyl or C 3 -C 7 -cycloalkyl that is substituted in one or more places in the same way or differently with hydroxy, C 1 -C 6 -alkoxy, halogen, phenyl, —NR 3 R 4 or phenyl, which itself can be substituted in one or more places in the same way or differently with halogen, hydroxy, C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy, halo-C 1 -C 6 -alkyl, or halo-C 1 -C 6 -alkoxy, or R 7 stands for phenyl, which itself can be substituted in one or more places in the same way or differently with halogen, hydroxy, C 1 -C 6 -alkyl or C 1 -C 6 -alkoxy, halo-C 1 -C 6 -alkyl, or halo-C 1 -C 6 -alkoxy,

R 8 , R 9 and

R 10 , in each case independently of one another, stand for hydrogen, hydroxy, C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkinyl, C 3 -C 10 -cycloalkyl, aryl, or heteroaryl or for C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkinyl or C 3 -C 10 -cycloalkyl that is substituted in one or more places in the same way or differently with hydroxy, halogen, C 1 -C 6 -alkoxy, C 1 -C 6 -alkylthio, amino, cyano, C 1 -C 6 -alkyl, —NH—(CH 2 ) n —C 3 -C 10 -cycloalkyl, C 3 -C 10 -cycloalkyl, C 1 -C 6 -hydroxyalkyl, C 2 -C 6 -alkenyl, C 2 -C 6 -alkinyl, C 1 -C 6 -alkoxy-C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy-C 1 -C 6 -alkoxy-C 1 -C 6 -alkyl, —NHC 1 -C 6 -alkyl, —N(C 1 -C 6 -alkyl) 2 , —SO(C 1 -C 6 -alkyl), —SO 2 (C 1 -C 6 -alkyl), C 1 -C 6 -alkanoyl, —CONR 3 R 4 , —COR 5 , C 1 -C 6 -alkylOAc, carboxy, aryl, heteroaryl, —(CH 2 ) n -aryl, —(CH 2 ) n -heteroaryl, phenyl-(CH 2 ) n —R 5 , —(CH 2 ) n PO 3 (R 5 ) 2 or with the group —R 6 or —NR 3 R 4 , and the phenyl, C 3 -C 10 -cycloalkyl, aryl, heteroaryl, —(CH 2 ) n -aryl and —(CH 2 ) n -heteroaryl itself optionally can be substituted in one or more places in the same way or differently with halogen, hydroxy, C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy or with the group —CF 3 or —OCF 3 , and the ring of C 3 -C 10 -cycloalkyl and the C 1 -C 10 -alkyl optionally can be interrupted by one or more nitrogen, oxygen and/or sulfur atoms and/or can be interrupted by one or more ═C═O groups in the ring and/or optionally one or more possible double bonds can be contained in the ring, and

n stands for 0-6,

as well as isomers, diastereomers, enantiomers and salts thereof that overcome known drawbacks.

Alkyl is defined in each case as a straight-chain or branched alkyl radical, such as, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, hexyl, heptyl, octyl, nonyl and decyl.

Alkoxy is defined in each case as a straight-chain or branched alkoxy radical, such as, for example, methyloxy, ethyloxy, propyloxy, isopropyloxy, butyloxy, isobutyloxy, sec-butyloxy, tert-butyloxy, pentyloxy, isopentyloxy, or hexyloxy.

Alkylthio is defined in each case as a straight-chain or branched alkylthio radical, such as, for example, methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, sec-butylthio, tert-butylthio, pentylthio, isopentylthio or hexylthio.

Cycloalkyl is defined in general as monocyclic alkyl rings, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecyl, but also bicyclic rings or tricyclic rings such as, for example, norbornyl, adamantanyl, etc.

The ring systems, in which optionally one or more possible double bonds can be contained in the ring, are defined as, for example, cycloalkenyls, such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, or cycloheptenyl, whereby the linkage can be carried out both to the double bond and to the single bonds.

If A and B, R 3 and R 4 , X and R 2 , in each case independently of one another, together form a C 3 -C 10 -cycloalkyl ring, which optionally can be interrupted by one or more heteroatoms, such as nitrogen atoms, oxygen atoms and/or sulfur atoms, and/or can be interrupted by one or more ═C═O groups in the ring and/or optionally one or more possible double bonds can be contained in the ring, the above-mentioned definitions, however, are also intended to include heteroaryl radical or heterocycloalkyl and heterocycloalkenyl.

Halogen is defined in each case as fluorine, chlorine, bromine or iodine.

The alkenyl substitutents in each case are straight-chain or branched, whereby, for example, the following radicals are meant: vinyl, propen-1-yl, propen-2-yl, but-1-en-1-yl, but-1-en-2-yl, but-2-en-1-yl, but-2-en-2-yl, 2-methyl-prop-2-en-1-yl, 2-methyl-prop-1-en-1-yl, but-1-en-3-yl, ethinyl, prop-1-in-1-yl, but-1-in-1-yl, but-2-in-1-yl, but-3-en-1-yl, and allyl.

Alkinyl is defined in each case as a straight-chain or branched alkinyl radical that contains 2-6, preferably 2-4, C atoms. For example, the following radicals can be mentioned: acetylene, propin-1-yl, propin-3-yl, but-1-in-1-yl, but-1-in-4-yl, but-2-in-1-yl, but-1-in-3-yl, etc.

The aryl radical in each case comprises 3-12 carbon atoms and in each case can be benzocondensed.

For example, there can be mentioned: cyclopropenyl, cyclopentadienyl, phenyl, tropyl, cyclooctadienyl, indenyl, naphthyl, azulenyl, biphenyl, fluorenyl, anthracenyl, etc.

The heteroaryl radical in each case comprises 3-16 ring atoms, and instead of the carbon can contain one or more heteroatoms that are the same or different, such as oxygen, nitrogen or sulfur, in the ring, and can be monocyclic, bicyclic, or tricyclic and in addition in each case can be benzocondensed.

For example, there can be mentioned:

Thienyl, furanyl, pyrrolyl, oxazolyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, thiadiazolyl, etc., and benzo derivatives thereof, such as, e.g., benzofuranyl, benzothienyl, benzoxazolyl, benzimidazolyl, indazolyl, indolyl, isoindolyl, etc.; or pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, etc., and benzo derivatives thereof, such as, e.g., quinolyl, isoquinolyl, etc., or azocinyl, indolizinyl, purinyl, etc., and benzo derivatives thereof; or quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, naphthyridinyl, pteridinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, xanthenyl, oxepinyl, etc.

Heterocycloalkyl stands for an alkyl ring that comprises 3-12 carbon atoms, which instead of the carbon contains one or more heteroatoms that are the same or different, such as, e.g., oxygen, sulfur or nitrogen.

As heterocycloalkyls, there can be mentioned, e.g.: oxiranyl, oxethanyl, aziridinyl, azetidinyl, tetrahydrofuranyl, pyrrolidinyl, dioxolanyl, imidazolidinyl, pyrazolidinyl, dioxanyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, trithianyl, quinuclidinyl, etc.

Heterocycloalkenyl stands for an alkyl ring that comprises 3-12 carbon atoms, which instead of the carbon contains one or more heteroatoms that are the same or different, such as, e.g., oxygen, sulfur or nitrogen, and which is partially saturated.

As heterocycloalkenyls, there can be mentioned, e.g.: pyran, thiin, dihydroacet, etc.

If an acid group is included, the physiologically compatible salts of organic and inorganic bases are suitable as salts, such as, for example, the readily soluble alkali and alkaline-earth salts, as well as N-methyl-glucamine, dimethyl-glucamine, ethyl-glucamine, lysine, 1,6-hexadiamine, ethanolamine, glucosamine, sarcosine, serinol, tris-hydroxy-methyl-amino-methane, aminopropane diol, Sovak base, and 1-amino-2,3,4-butanetriol.

If a basic group is included, the physiologically compatible salts of organic and inorganic acids are suitable, such as hydrochloric acid, sulfuric acid, phosphoric acid, citric acid, tartaric acid, i.a.

Those compounds of general formula (I) in which

R 1 stands for hydrogen, halogen, C 1 -C 6 -alkyl, nitro, or for the group —COR 5 , —OCF 3 , —(CH 2 ) n R 5 , —S—CF 3 or —SO 2 CF 3 , R 2 stands for C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkinyl, or C 3 -C 10 -cycloalkyl or for C 1 -C 10 -alkyl, C 2 -C 10 -alkenyl, C 2 -C 10 -alkinyl, or C 3 -C 10 -cycloalkyl that is substituted in one or more places in the same way or differently with hydroxy, halogen, C 1 -C 6 -alkoxy, C 1 -C 6 -alkylthio, amino, cyano, C 1 -C 6 -alkyl, —NH—(CH 2 ) n —C 3 -C 10 -cycloalkyl, C 3 -C 10 -cycloalkyl, C 1 -C 6 -hydroxyalkyl, C 2 -C 6 -alkenyl, C 2 -C 6 -alkinyl, C 1 -C 6 -alkoxy-C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy-C 1 -C 6 -alkoxy-C 1 -C 6 -alkyl, —NHC 16-alkyl, —N(C 1 -C 6 -alkyl) 2 , —SO(C 1 -C 6 -alkyl), —SO 2 (C 1 -C 6 -alkyl), C 1 -C 6 -alkanoyl, —CONR 3 R 4 , —COR 5 , C 1 -C 6 -alkylOAc, carboxy, aryl, heteroaryl, —(CH 2 ) n -aryl, —(CH 2 ) n -heteroaryl, phenyl-(CH 2 ) n —R 5 , —(CH 2 ) n PO 3 (5) 2 or with the group —R 6 or —NR 3 R 4 , and the phenyl, C 3 -C 10 -cycloalkyl, aryl, heteroaryl, —(CH 2 ) n -aryl and —(CH 2 ) n -heteroaryl itself optionally can be substituted in one or more places in the same way or differently with halogen, hydroxy, C 1 -C 6 -alkyl, C 1 -C 6 alkoxy, heteroaryl, benzoxy or with the group —CF 3 or —OCF 3 , and the ring of the C 3 -C 10 -cycloalkyl and the C 1 -C 10 -alkyl optionally can be interrupted by one or more nitrogen, oxygen and/or sulfur atoms and/or can be interrupted by one or more ═C═O groups in the ring and/or optionally one or more possible double bonds can be contained in the ring, or R 2 stands for the group

X stands for oxygen or for the group —NH—, —N(C 1 -C 3 -alkyl) or for —OC 3 -C 10 -cycloalkyl, which can be substituted in one or more places in the same way or differently with a heteroaromatic compound,

or

whereby the phenyl itself can be substituted in one or more places in the same way or differently with halogen, trifluoromethyl, C 1 -C 6 -alkyl, C 1 -C 6 -alkoxy or with the group —SO 2 NR 3 R 4 , or for the group —(CH 2 ) n NR 3 R 4 , —CNHNH 2 or —NR 3 R 4 or for

as well as isomers, enantiomers, diastereomers, and salts thereof, are especially effective.

Those compounds of general formula I in which

R 1 stands for hydrogen, halogen, C 1 -C 3 -alkyl, or for the group —(CH 2 ) n R 5 , R 2 stands for —CH(CH 3 )—(CH 2 ) n —R 5 , —CH—(CH 2 OH) 2 , —(CH 2 ) n R 7 , —CH(C 3 H 7 )—(CH 2 ) n —R 5 , —CH(C 2 H 5 )—(CH 2 ) n —R 5 , —CH 2 —CN, —CH(CH 3 )COCH 3 , —CH(CH 3 )—C(OH)(CH 3 ) 2 , —CH(CH(OH)CH 3 )OCH 3 , CH(C 2 H 5 )CO—R 5 , C 2 -C 4 -alkinyl, —(CH 2 ) n —COR 5 , —(CH 2 ) n —CO—C 1 -C 6 -alkyl, —(CH 2 ) n —C(OH)(CH 3 )-phenyl, —CH(CH 3 )—C(CH 3 )—R 5 , —CH(CH 3 )—C(CH 3 )(C 2 H 5 )—R 5 , —CH(OCH 3 )—CH 2 —R 5 , —CH 2 —CH(OH)—R 5 , —CH(OCH 3 )—CHR 5 —CH 3 , —CH(CH 3 )—CH(OH)—CH 2 —CH═CH 2 , —CH(C 2 H 5 )—CH(OH)—(CH 2 ) n —CH 3 , —CH(CH 3 )—CH(OH)—(CH 2 ) n —CH 3 , —CH(CH 3 )—CH(OH)—CH(CH 3 ) 2 , (CH 2 OAC) 2 , —(CH 2 ) n —R 6 , —(CH 2 ) n —(CF 2 ) n —CF 3 , —CH(CH 2 ) n —R 5 ) 2 , —CH(CH 3 )—CO—NH 2 , —CH(CH 2 OH)-phenyl, —CH(CH 2 OH)—CH(OH)—(CH 2 ) n R 5 , —CH(CH 2 OH)—CH(OH)-phenyl, —CH(CH 2 OH)—C 2 H 4 —R 5 , —(CH 2 ) n —C≡C(CH 3 )═CH—COR 5 , —CH(Ph)-(CH 2 ) n —R 5 , —(CH 2 ) n —COR 5 , —(CH 2 ) n PO 3 (R 5 ) 2 , —(CH 2 ) n —COR 5 , —CH((CH 2 ) n OR 5 )CO—R 5 , —(CH 2 ) n CONHCH((CH 2 ) n R 5 ) 2 , —(CH 2 ) n NH—COR 5 , —CH(CH 2 ) n R 5 —(CH 2 ) n C 3 -C 10 -cycloalkyl, —(CH 2 ) n —C 3 -C 10 -cycloalkyl, C 3 -C 10 -cycloalkyl; C 1 -C 6 -alkyl, C 3 -C 10 -cycloalkyl, —(CH 2 ) n —O—(CH 2 ) n —R 5 , —(CH 2 ) n —NR 3 R 4 that is optionally substituted in one or more places in the same way or differently with hydroxy, C 1 -C 6 -alkyl or the group —COONH(CH 2 ) n CH 3 or —NR 3 R 4 , —CH(C 3 H 7 )—(CH 2 ) n —OC(O)—(CH 2 ) n —CH 3 , —(CH 2 ) n —R 5 , —C(CH 3 ) 2 —(CH 2 ) n —R 5 , —C(CH 2 ) n (CH 3 )—(CH 2 ) n R 5 , —C(CH 2 ) n —(CH 2 ) n R 5 , —CH(t-butyl)-(CH 2 ) n —R 5 , —CCH 3 (C 3 H 7 )—(CH 2 ) n R 5 , —CH(C 3 H 7 )—(CH 2 ) n —R 5 , —CH(C 3 H 7 )—COR 5 , —CH(C 3 H 7 )—(CH 2 ) n —OC(O)—NH—Ph,

—CH((CH 2 ) n (C 3 H 7 ))—(CH 2 )R 5 , —CH(C 3 H 7 )—(CH 2 ) n —OC(O)—NH—Ph(OR 5 ) 3 , —NR 3 R 4 , —NH—(CH 2 ) n —NR 3 R 4 , R—(CH 2 ) n —C*H—CH(R 5 )—(CH 2 ) n —R 5 , —(CH 2 ) n —CO—NH—(CH 2 ) n —CO—R 5 , —OC(O)NH—C 1 -C 6 -alkyl or —(CH 2 ) n —CO—NH—(CH 2 ) n —CH—((CH 2 ) n R 5 ) 2 ,

or for the group —(CH 2 ) n NR 3 R 3 R 4 , —CNHNH 2 or —NR 3 R 4 or for

as well as isomers, diastereomers, enantiomers and salts thereof, have proven quite especially effective.

The compounds according to the invention essentially inhibit cyclin-dependent kinases, upon which is based their action, for example, against cancer, such as solid tumors and leukemia; auto-immune diseases such as psoriasis, alopecia, and multiple sclerosis, chemotherapy-induced alopecia and mucositis; cardiovascular diseases such as stenoses, arterioscleroses and restenoses; infectious diseases, such as, e.g., by unicellular parasites, such as trypanosoma, toxoplasma or plasmodium, or produced by fungi; nephrological diseases, such as, e.g., glomerulonephritis, chronic neurodegenerative diseases, such as Huntington's disease, amyotropic lateral sclerosis, Parkinson's disease, AIDS dementia and Alzheimer's disease; acute neurodegenerative diseases, such as ischemias of the brain and neurotraumas; viral infections, such as, e.g., cytomegalic infections, herpes, Hepatitis B and C, and HIV diseases.

The eukaryotic cell division ensures the duplication of the genome and its distribution to the daughter cells by passing through a coordinated and regulated sequence of events. The cell cycle is divided into four successive phases: the G1 phase represents the time before the DNA replication, in which the cell grows and is sensitive to external stimuli. In the S phase, the cell replicates its DNA, and in the G2 phase, preparations are made for entry into mitosis. In mitosis (M phase), the replicated DNA separates, and cell division is completed.

The cyclin-dependent kinases (CDKs), a family of serine/threonine kinases, whose members require the binding of a cyclin (Cyc) as a regulatory subunit in order for them to activate, drive the cell through the cell cycle. Different CDK/Cyc pairs are active in the various phases of the cell cycle. CDK/Cyc pairs that are important to the basic function of the cell cycle are, for example, CDK4(6)/CycD, CDK2/CycE, CDK2/CycA, CDK1/CycA and CDK1/CycB. Some members of the CDK enzyme family have a regulatory function by influencing the activity of the above-mentioned cell cycle CDKs, while no specific function could be associated with other members of the CDK enzyme family. One of the latter, CDK5, is distinguished in that it has an atypical regulatory subunit (p35) that deviates from the cyclins, and its activity is highest in the brain.

The entry into the cell cycle and the passage through the “restriction points,” which marks the independence of a cell from further growth signals for the completion of the cell division that has begun, are controlled by the activity of the CDK4(6)/CycD and CDK2/CycE complexes. The essential substrate of these CDK complexes is the retinoblastoma protein (Rb), the product of the retinoblastoma tumor suppressor gene. Rb is a transcriptional co-repressor protein. In addition to other, still largely little understood mechanisms, Rb binds and inactivates transcription factors of the E2F type and forms transcriptional repressor complexes with histone-deacetylases (HDAC) (Zhang, H. S. et al. (2000). Exit from G1 and S Phase of the Cell Cycle is Regulated by Repressor Complexes Containing HDAC-Rb-hSWI/SNF and Rb-hSWI/SNF. Cell 101, 79-89). By the phosphorylation of Rb by CDKs, bonded E2F transcription factors are released and result in transcriptional activation of genes, whose products are required for the DNA synthesis and the progression through the S phase. In addition, the Rb-phosphorylation brings about the breakdown of the Rb-HDAC complexes, by which additional genes are activated. The phosphorylation of Rb by CDK's is to be treated as equivalent to exceeding the “restriction points.” For the progression through the S phase and its completion, the activity of the CDK2/CycE and CDK2/CycA complexes is necessary, e.g., the activity of the transcription factors of the E2F type is turned off by means of phosphorylation by CDK2/CycA as soon as the cells are entered into the S phase. After replication of DNA is complete, the CDK1 in the complex with CycA or CycB controls the entry into and the passage through phases G2 and M ( FIG. 1 ).

According to the extraordinary importance of the cell-division cycle, the passage through the cycle is strictly regulated and controlled. The enzymes that are necessary for the progression through the cycle must be activated at the correct time and are also turned off again as soon as the corresponding phase is passed. Corresponding control points (“checkpoints”) stop the progression through the cell cycle if DNA damage is detected, or the DNA replication or the creation of the spindle device is not yet completed.

The activity of the CDKs is controlled directly by various mechanisms, such as synthesis and degradation of cyclins, complexing of the CDKs with the corresponding cyclins, phosphorylation and dephosphorylation of regulatory threonine and tyrosine radicals, and the binding of natural inhibitory proteins. While the amount of protein of the CDKs in a proliferating cell is relatively constant, the amount of the individual cyclins oscillates with the passage through the cycle. Thus, for example, the expression of CycD during the early G1 phase is stimulated by growth factors, and the expression of CycE is induced after the “restriction points” are exceeded by the activation of the transcription factors of the E2F type. The cyclins themselves are degraded by the ubiquitin-mediated proteolysis. Activating and inactivating phosphorylations regulate the activities of the CDKs, for example phosphorylate CDK-activating kinases (CAKs) Thr160/161 of the CDK1, while, by contrast, the families of Wee1/Myt1 inactivate kinases CDK1 by phosphorylation of Thr14 and Tyr15. These inactivating phosphorylations can be destroyed in turn by cdc25 phosphatases. The regulation of the activity of the CDK/Cyc complexes by two families of natural CDK inhibitor proteins (CKIs), the protein products of the p21 gene family (p21, p27, p57) and the p16 gene family (p15, p16, p18, p19) is very significant. Members of the p21 family bind to cyclin complexes of CDKs 1,2,4,6, but inhibit only the complexes that contain CDK1 or CDK2. Members of the p16 family are specific inhibitors of the CDK4 and CDK6 complexes.

The plane of control point regulation lies above this complex direct regulation of the activity of the CDKs. Control points allow the cell to track the orderly sequence of the individual phases during the cell cycle. The most important control points lie at the transition from G1 to S and from G2 to M. The G1 control point ensures that the cell does not initiate any DNA synthesis unless it has proper nutrition, interacts correctly with other cells or the substrate, and its DNA is intact. The G2/M control point ensures the complete replication of DNA and the creation of the mitotic spindle before the cell enters into mitosis. The G1 control point is activated by the gene product of the p53 tumor suppressor gene. p53 is activated after detection of changes in metabolism or the genomic integrity of the cell and can trigger either a stopping of the cell cycle progression or apoptosis. In this case, the transcriptional activation of the expression of the CDK inhibitor protein p21 by p53 plays a decisive role. A second branch of the G1 control point comprises the activation of the ATM and Chk1 kinases after DNA damage by UV light or ionizing radiation and finally the phosphorylation and the subsequent proteolytic degradation of the cdc25A phosphatase (Mailand, N. et al. (2000). Rapid Destruction of Human cdc25A in Response to DNA Damage. Science 288, 1425-1429). A shutdown of the cell cycle results from this, since the inhibitory phosphorylation of the CDKs is not removed. After the G2/M control point is activated by damage of the DNA, both mechanisms are involved in a similar way in stopping the progression through the cell cycle.

The loss of the regulation of the cell cycle and the loss of function of the control points are characteristics of tumor cells. The CDK-Rb signal path is affected by mutations in over 90% of human tumor cells. These mutations, which finally result in inactivating phosphorylation of the RB, include the over-expression of D- and E-cyclins by gene amplification or chromosomal translocations, inactivating mutations or deletions of CDK inhibitors of the p16 type, as well as increased (p27) or reduced (CycD) protein degradation. The second group of genes, which are affected by mutations in tumor cells, codes for components of the control points. Thus p53, which is essential for the G1 and G2/M control points, is the most frequently mutated gene in human tumors (about 50%).

In tumor cells that express p53 without mutation, it is often inactivated because of a greatly increased protein degradation. In a similar way, the genes of other proteins that are necessary for the function of the control points are affected by mutations, for example ATM (inactivating mutations) or cdc25 phosphatases (over-expression).

Convincing experimental data indicate that CDK2/Cyc complexes occupy a decisive position during the cell cycle progression: (1) Both dominant-negative forms of CDK2, such as the transcriptional repression of the CDK2 expression by anti-sense oligonucleotides, produce a stopping of the cell cycle progression. (2) The inactivation of the CycA gene in mice is lethal. (3) The disruption of the function of the CDK2/CycA complex in cells by means of cell-permeable peptides resulted in tumor cell-selective apoptosis (Chen, Y. N. P. et al. (1999). Selective Killing of Transformed Cells by Cyclin/Cyclin-Dependent Kinase 2 Antagonists. Proc. Natl. Acad. Sci. USA 96, 4325-4329).

Changes of the cell cycle control play a role not only in carcinoses. The cell cycle is activated by a number of viruses, both by transforming viruses as well as by non-transforming viruses, to make possible the replication of viruses in the host cell. The false entry into the cell cycle of normally post-mitotic cells is associated with various neurodegenerative diseases. The mechanisms of the cell cycle regulation, their changes in diseases and a number of approaches to develop inhibitors of the cell cycle progression and especially the CDKs were already described in a detailed summary in several publications (Sielecki, T. M. et al. (2000). Cyclin-Dependent Kinase Inhibitors: Useful Targets in Cell Cycle Regulation. J. Med. Chem. 43, 1-18; Fry, D. W. & Garrett, M. D. (2000). Inhibitors of Cyclin-Dependent Kinases as Therapeutic Agents for the Treatment of Cancer. Curr. Opin. Oncol. Endo. Metab. Invest. Drugs 2, 40-59; Rosiania, G. R. & Chang, Y. T. (2000). Targeting Hyperproliferative Disorders with Cyclin-Dependent Kinase Inhibitors. Exp. Opin. Ther. Patents 10, 215-230; Meijer, L. et al. (1999). Properties and Potential Applications of Chemical Inhibitors of Cyclin-Dependent Kinases. Pharmacol. Ther. 82, 279-284; Senderowicz, A. M. & Sausville, E. A. (2000). Preclinical and Clinical Development of Cyclin-Dependent Kinase Modulators. J. Natl. Cancer Inst. 92, 376-387).

To use the compounds according to the invention as pharmaceutical agents, the latter are brought into the form of a pharmaceutical preparation, which in addition to the active ingredient for enteral or parenteral administration contains suitable pharmaceutical, organic or inorganic inert carrier materials, such as, for example, water, gelatin, gum arabic, lactose, starch, magnesium stearate, talc, vegetable oils, polyalkylene glycols, etc. The pharmaceutical preparations can be present in solid form, for example as tablets, coated tablets, suppositories, or capsules, or in liquid form, for example as solutions, suspensions, or emulsions. Moreover, they optionally contain adjuvants, such as preservatives, stabilizers, wetting agents or emulsifiers; salts for changing the osmotic pressure or buffers. These pharmaceutical preparations are also subjects of this invention.

For parenteral administration, especially injection solutions or suspensions, especially aqueous solutions of active compounds in polyhydroxyethoxylated castor oil, are suitable.

As carrier systems, surface-active adjuvants such as salts of bile acids or animal or plant phospholipids, but also mixtures thereof, as well as liposomes or their components can also be used.

For oral administration, especially tablets, coated tablets or capsules with talc and/or hydrocarbon vehicles or binders, such as, for example, lactose, corn or potato starch, are suitable. The administration can also be carried out in liquid form, such as, for example, as a juice, to which optionally a sweetener is added.

Enteral, parenteral and oral administrations are also subjects of this invention.

The dosage of the active ingredients can vary depending on the method of administration, age and weight of the patient, type and severity of the disease to be treated and similar factors. The daily dose is 0.5-1000 mg, preferably 50-200 mg, whereby the dose can be given as a single dose to be administered once or divided into two or more daily doses.

Subjects of this invention also include the use of compounds of general formula I for the production of a pharmaceutical agent for treating cancer, auto-immune diseases, cardiovascular diseases, chemotherapy agent-induced alopecia and mucositis, infectious diseases, nephrological diseases, chronic and acute neurodegenerative diseases and viral infections, whereby cancer is defined as solid tumors and leukemia; auto-immune diseases are defined as psoriasis, alopecia and multiple sclerosis; cardiovascular diseases are defined as stenoses, arterioscleroses and restenoses; infectious diseases are defined as diseases that are caused by unicellular parasites; nephrological diseases are defined as glomerulonephritis; chronic neurodegenerative diseases are defined as Huntington's disease, amyotrophic lateral sclerosis, Parkinson's disease, AIDS dementia and Alzheimer's disease; acute neurodegenerative diseases are defined as ischemias of the brain and neurotraumas; and viral infections are defined as cytomegalic infections, herpes, hepatitis B or C, and HIV diseases.

Subjects of this invention also include pharmaceutical agents for treating the above-cited diseases, which contain at least one compound according to general formula I, as well as pharmaceutical agents with suitable formulation substances and vehicles.

The compounds of general formula I according to the invention are, i.a., excellent inhibitors of the cyclin-dependent kinases, such as CDK1, CDK2, CDK3, CDK4, CDK5, CDK6, CDK7, CDK8 and CDK9, as well as the glycogen-synthase-kinase (GSK-3β).

If the production of the starting compounds is not described, these compounds are known or can be produced analogously to known compounds or to processes that are described here. It is also possible to perform all reactions that are described here in parallel reactors or by means of combinatory operating procedures.

The isomer mixtures can be separated into the enantiomers or E/Z isomers according to commonly used methods, such as, for example, crystallization, chromatography or salt formation.

The production of the salts is carried out in the usual way by a solution of the compound of formula I being mixed with the equivalent amount of or excess base or acid, which optionally is in solution, and the precipitate being separated or the solution being worked up in the usual way.

The following examples explain the production of the compounds according to the invention, without the scope of the claimed compounds being limited to these examples.

The compounds of general formula I according to the invention can be produced according to the following general diagrams of the process:

MH + 429 (29%)

371 (61%)

289 (91%)

Similarly produced are also the following compounds: