US20110086842A1

US20110086842A1 - Pyrimidines as PLK inhibitors

Info

Publication number: US20110086842A1
Application number: US12/972,976
Authority: US
Inventors: Heinz Stadtmueller; Harald Engelhardt; Martin Steegmaier; Anke Baum; Ulrich Guertler; Andreas Schoop; Jens Juergen Quant; Flavio Solca; Rudolf Hauptmann; Ulrich Reiser; Stephan Karl Zahn; Lars Herfurth
Original assignee: Boehringer Ingelheim International GmbH
Current assignee: Boehringer Ingelheim International GmbH
Priority date: 2004-08-20
Filing date: 2010-12-20
Publication date: 2011-04-14
Also published as: NZ553632A; EP1781640A1; ZA200700084B; CA2573371A1; JP5060294B2; US20090163465A1; PL1781640T3; CN101044137A; ES2532611T3; AU2005276500A1; KR20070048757A; MX2007002055A; CA2573371C; WO2006021544A1; US20090149438A1; TW200619218A; RU2007109870A; DK1781640T3; IL181421A0; EP1781640B1

Abstract

The present invention encompasses compounds of general formula (1),

wherein
A, W, X, Y, Z, Ra, Rb, Rc, R1 and R3 are defined as in claim 1, which are suitable for the treatment of diseases characterised by excessive or abnormal cell proliferation, and the use thereof for preparing a pharmaceutical composition having the above-mentioned properties.

Description

The present invention relates to new pyrimidines of general formula (1),
wherein the groups A, W, X, Y, Z, R^a, R^b, R^c, R¹and R³have the meanings given in the claims and description, the isomers thereof, processes for preparing these pyrimidines and their use as pharmaceutical compositions.

BACKGROUND TO THE INVENTION

Tumour cells wholly or partly elude regulation and control by the body and are characterised by uncontrolled growth. This is due on the one hand to the loss of control proteins such as for example Rb, p16, p21 and p53 and also to the activation of so-called accelerators of the cell cycle, the cyclin-dependent kinases.
Studies in model organisms such as Schizosaccharomyces pombe, Drosophila melanogaster or Xenopus laevis as well as investigations in human cells have shown that the transition from the G2 phase to mitosis is regulated by the CDK1/cyclin B kinase (Nurse 1990, Nature 344: 503-508). This kinase, which is also known as “mitosis promoting factor” (MPF), phosphorylates and regulates a plurality of proteins, such as e.g. nuclear lamina, kinesin-like motor proteins, condensins and Golgi Matrix Proteins, which play an important part in the breakdown of the nuclear coat, in centrosome separation, the structure of the mitotic spindle apparatus, chromosome condensation and breakdown of the Golgi apparatus (Nigg. E. 2001, Nat Rev Mol Cell Biol. 2(1):21-32). A murine cell line with a temperature-sensitive CDK-1 kinase mutant shows a rapid breakdown in CDK-1 kinase after temperature increase and a subsequent arrest in the G2/M phase (Th'ng et al. 1990, Cell. 63(2):313-24). The treatment of human tumour cells with inhibitors against CDK1/cyclin B, such as e.g. butyrolactone, leads to an arrest in the G2/M phase and subsequent apoptosis (Nishio, et al. 1996, Anticancer Res. 16(6B):3387-95).
Moreover, the protein kinase Aurora B has also been described as having an essential function during entry into mitosis. Aurora B phosphorylates histone H3 on Ser 10 and thereby initiates chromosome condensation (Hsu et al. 2000, Cell 102:279-91). A specific cell cycle arrest in the G2/M phase may, however, also be initiated e.g. by inhibition of specific phosphatases such as e.g. Cdc25C (Russell and Nurse 1986, Cell 45:145-53). Yeasts with a defective Cdc25 gene arrest in the G2 phase, whereas overexpression of Cdc25 leads to premature entry into the mitosis phase (Russell and Nurse, 1987, Cell 49:559-67). Moreover, an arrest in the G2/M phase may also be initiated by inhibition of specific motor proteins, the so-called kinesins such as for example Eg5 (Mayer et al. 1999, Science 286:971-4), or by microtubuli stabilising or destabilising agents (e.g. colchicin, taxol, etoposide, vinblastine, vincristine) (Schiff and Horwitz 1980, Proc Natl Acad Sci USA 77:1561-5).
In addition to the cyclin-dependent and Aurora kinases the so-called polo-like kinases, a small family of serine/threonine kinases, also play an important role in the regulation of the eukaryotic cell cycle. Up till now the polo-like kinases PLK-1, PLK-2, PLK-3 and PLK-4 have been described in the literature. PLK-1 in particular has been found to play a central role in the regulation of the mitosis phase. PLK-1 is responsible for the maturation of the centrosomes, for the activation of phosphatase Cdc25C, as well as for the activation of the Anaphase Promoting Complex (Glover et al. 1998, Genes Dev. 12:3777-87; Qian et al. 2001, Mol Biol Cell. 12:1791-9). The injection of PLK-1 antibodies leads to a G2 arrest in untransformed cells, whereas tumour cells arrest during the mitosis phase (Lane and Nigg 1996, J. Cell Biol. 135:1701-13). Overexpression of PLK-1 has been demonstrated in various types of tumour, such as non-small-cell carcinoma of the lung, plate epithelial carcinoma, breast and colorectal carcinoma (Wolf et al. 1997, Oncogene 14:543-549; Knecht et al. 1999, Cancer Res. 59:2794-2797; Wolf et al. 2000, Pathol. Res. Pract. 196:753-759; Takahashi et al. 2003, Cancer Sci. 94:148-52). Therefore, this category of proteins also presents an interesting point of attack for therapeutic intervention in proliferative diseases (Liu and Erikson 2003, Proc Natl Acad Sci USA 100:5789-5794).
Pyrimidines are generally known as inhibitors of kinases. Thus, for example, pyrimidines are described as an active component with an anticancer activity in International Patent Application WO 00/53595, which describes the use of 2,4,5-substituted pyrimidines with a heterocyclic group in the 4-position and an anilino group in the 2 position, which in turn comprises a side chain with the length of at least one n-propyl group.
Moreover, International Patent Application WO 00/39101 describes the use of 2,4,5-substituted pyrimidines as compounds with an anticancer activity which are linked in the 2- and 4-position with an aromatic or heteroaromatic ring, at least one of which comprises a side chain with the length of at least one n-propyl group.
International Patent Application WO 97/19065 further proposes the use of 2,4,5-substituted pyrimidines with a 3,4-dialkoxyanilino group in position 2 as kinase inhibitors.
International Patent Application WO 02/04429 describes 2,4,5-substituted pyrimidines with a cyano group in position 5 and their cell cycle inhibiting effect.
International Patent Application WO 03/063794 describes the use of 2,4-pyrimidinediamines as inhibitors of the IgE and/or IgG receptor signal cascade.
Antiviral 2,4,5-substituted pyrimidines, wherein the groups R^cand R^dform a heteroaromatic five-membered ring at the nitrogen of the 4-position, are known from International Patent Application WO 99/41253.
2,4,5-substituted pyrimidines which carry (hetero)aryls in position 2 and 4 (WO00/27825) and also 2,4,5-substituted pyrimidines which carry a (hetero)aryl group functionalised with a nitrile group in position 2 or 4 (EP 0 945 443 A1) are described as having an antiviral activity.
The resistance of many types of tumour demands that new drugs be developed to fight the tumours. The aim of the present invention is therefore to indicate new active substances which may be used for the prevention and/or treatment of diseases characterised by excessive or anomalous cell proliferation.

DETAILED DESCRIPTION OF THE INVENTION

It has now been found that, surprisingly, compounds of general formula (I), wherein the groups A, W, X, Y, R^a, R^b, R^c, R¹, R²and R³are defined as hereinafter, act as inhibitors of specific cell cycle kinases. Thus, the compounds according to the invention may be used for example for the treatment of diseases associated with the activity of specific cell cycle kinases and characterised by excessive or anomalous cell proliferation.
The present invention relates to compounds of general formula (1)
wherein

W denotes N or C—R²,
X denotes —NR^1a, O or S,
Y denotes CH or N,
Z denotes hydrogen, halogen, —NO₂, C_1-3alkyl, C_2-3alkenyl, C_2-3alkynyl, halogen-C_1-3alkyl, —COH, —C(═O)—C_1-3alkyl, —C(═O)—C_2-3alkenyl, —C(═O)—C_2-3alkynyl, —C(═O)C_1-3alkyl-halogen or pseudohalogen;
A is selected from the formulae (i), (ii) or (iii)

Q₁denotes mono- or bicyclic aryl compounds;
B¹, B², B³and B⁴in each case independently of one another denote C—R^gR^h, N—Rⁱ, O or S, while adjacent B¹-B⁴in each case do not represent —O—;
R¹and R^1aeach independently of one another denote hydrogen or methyl,
R²denotes a group selected from among hydrogen, halogen, —OR⁴, —C(═O)R⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —C═NRⁱ, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵and pseudohalogen, or an optionally mono- or polysubstituted group selected from among C_1-6alkyl, C_2-6alkenyl, C_2-6alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen;
R^a, R^b, R^c, R^d, R^e, R^f, R^gand R^hin each case independently of one another denote a group selected from among hydrogen, halogen, ═O, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —C═NRⁱ, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen; group functionalised with a nitrile group in position 2 or 4 (EP 0 945 443 A1) are described as having an antiviral activity.

The resistance of many types of tumour demands that new drugs be developed to fight the tumours. The aim of the present invention is therefore to indicate new active substances which may be used for the prevention and/or treatment of diseases characterised by excessive or anomalous cell proliferation.

DETAILED DESCRIPTION OF THE INVENTION

It has now been found that, surprisingly, compounds of general formula (I), wherein the groups A, W, X, Y, R^a, R^b, R^e, R¹, R²and R³are defined as hereinafter, act as inhibitors of specific cell cycle kinases. Thus, the compounds according to the invention may be used for example for the treatment of diseases associated with the activity of specific cell cycle kinases and characterised by excessive or anomalous cell proliferation.
The present invention relates to compounds of general formula (1)
wherein

W denotes N or C—R²,
X denotes —NR^1a, O or S,
Y denotes CH or N,

R⁴, R⁵and R⁶each independently of one another denote hydrogen or a group selected from among optionally mono- or polysubstituted C_1-5-alkyl, C_2-5alkenyl, C_2-5alkynyl, C_3-10cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among C_3-10-cycloalkyl, aryl, heterocyclyl, heteroaryl, halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
L denotes a bond or a group selected from among optionally mono- or polysubstituted C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
Q₂and Q₃independently of one another denote a bond or a group selected from among optionally mono- or polysubstituted C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl while the substituent(s) may be identical or different and are selected from among halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
R⁷denotes hydrogen or a group selected from among optionally mono- or polysubstituted C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸COR⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR^B, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
R⁸, R⁹and R¹⁰each independently of one another denote hydrogen or a group selected from among optionally substituted C_1-8-alkyl, C_2-8-alkenyl, C_2-8-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, methyl, ethyl, amino, methylamino, dimethylamino, —OH and pseudohalogen;
optionally in the form of the tautomers, racemates, enantiomers, diastereomers and mixtures thereof, and optionally the pharmacologically acceptable acid addition salts thereof.

In one aspect the invention relates to compounds of general formula (I) wherein

W denotes C—R²and the other groups are as hereinbefore defined.

In another aspect the invention relates to compounds of general formula (I), wherein

X denotes —NR^1aor oxygen,
R¹and R^1adenote hydrogen;
R³denotes formula (Iv) or (x),

and the other groups are as hereinbefore defined.
In another aspect the invention relates to compounds of general formula (I), wherein

Y denotes CH and
Q₁denotes monocyclic aryl compounds
and the other groups are as hereinbefore defined.

In one aspect the invention relates to compounds of general formula (I), wherein

R^cdenotes a group selected from among hydrogen, —F, —Cl, methyl and ethyl and the other groups are as hereinbefore defined.

R^aand R^beach independently of one another denote hydrogen or fluorine;
- or an optionally mono- or polysubstituted group selected from among C_1-2-alkyl, C₂-alkenyl, C₂-alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among hydrogen, halogen, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁵, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴, —SO₂NR⁴R⁵, —OSO₂NR⁴R⁵and pseudohalogen
  and the other groups are as hereinbefore defined.

In another aspect the invention also relates to compounds of general formula (I), wherein
R^aand R^bdenote hydrogen or fluorine and the other groups are as hereinbefore defined.
The invention also includes compounds of general formula (I), wherein

Z denotes halogen-C_1-3-alkyl, —COH, —C(═O)—C_1-3-alkyl, —C(═O)—C_2-3-alkenyl, —C(═O)—C_2-3-alkynyl, —C(═O)C_1-3-alkyl-halogen and pseudohalogen
and the other groups are as hereinbefore defined.

In one aspect the invention relates to compounds of general formula (I), or the pharmaceutically active salts thereof, as pharmaceutical compositions.
In an essential aspect the invention relates to compounds of general formula (I), or the pharmaceutically active salts thereof, for use as pharmaceutical compositions with an antiproliferative activity.
Moreover the invention includes compounds of general formula (I), or the pharmaceutically active salts thereof, for use as pharmaceutical compositions with an antiproliferative activity with a selective kinase-inhibiting mechanism of activity.
In one aspect the invention relates to the use of compounds of general formula (I), or the pharmaceutically active salts thereof, for preparing a pharmaceutical composition with an antiproliferative activity with a PLK inhibiting mechanism of activity.
In another aspect the invention relates to pharmaceutical preparations, containing as active substance one or more compounds of general formula (I), or the physiologically acceptable salts thereof, optionally in conjunction with conventional excipients and/or carriers.
In another aspect the invention relates to the use of one or more compounds of general formula (I) for preparing a pharmaceutical composition for the treatment and/or prevention of cancer, infections, inflammatory and autoimmune diseases.
In another aspect the invention relates to a pharmaceutical preparation containing at least one compound of general formula (I)
wherein

W denotes N or C—R²,
X denotes —NR^1a, O or S,
Y denotes CH or N,
Z denotes hydrogen, halogen, —NO₂, C_1-3-alkyl, C_2-3-alkenyl, C_2-3-alkynyl, halogen-C_1-3-alkyl, —COH, —C(═O)—C_1-3-alkyl, —C(═O)—C_2-3-alkenyl, —C(═O)—C_2-3-alkynyl, —C(═O)C_1-3-alkyl-halogen and pseudohalogen;
A is selected from the formulae (i), (ii) or (iii)

Q₁denotes mono- or bicyclic aryl compounds;
B¹, B², B³and B⁴in each case independently of one another represent C—R^gR^h, N—Rⁱ, O or S, while adjacent B¹-B⁴in each case do not denote —O—;
R¹and R^1aeach independently of one another denote hydrogen or methyl,
R²denotes a group selected from among hydrogen, halogen, —OR⁴, —C(═O)R⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —C═NRⁱ, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵and pseudohalogen, or an optionally mono- or polysubstituted group selected from among C_1-6-alkyl, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen;
R^a, R^b, R^e, R^d, R^e, R^f, R^gand R^hin each case independently of one another denote a group selected from among hydrogen, halogen, ═O, —NO₂, —OR⁴, —C(═O)R⁴, —C═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —C═NRⁱ, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen;
- or an optionally mono- or polysubstituted group selected from among C_1-6-alkyl, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, R⁸, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen; and optionally the R^gand R^hlocated at the same or at adjacent C atoms may be attached in any combination to a common saturated or partially unsaturated 3-5-membered alkyl bridge which may contain one to two heteroatoms;
Rⁱdenotes a group selected from among hydrogen, ═O)—OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶or an optionally mono- or polysubstituted group selected from among C_1-6-alkyl, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, R⁸, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen; and optionally the Ri groups located at adjacent N atoms may be joined together or to R^gand R^hlocated at adjacent C atoms in any combination with a common saturated or partially unsaturated 3-5-membered alkyl bridge which may contain one to two heteroatoms;
R³is selected from the formulae (iv)-(x),

R⁴, R⁵and R⁶each independently of one another denote hydrogen or a group selected from among optionally mono- or polysubstituted C_1-5-alkyl, C_2-5-alkenyl, C_2-5-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among C_3-10-cycloalkyl, aryl, heterocyclyl, heteroaryl, halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
L denotes a bond or a group selected from among optionally mono- or polysubstituted C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR^B, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
Q₂and Q₃independently of one another denote a bond or a group selected from among optionally mono- or polysubstituted C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
R⁷denotes hydrogen or a group selected from among optionally mono- or polysubstituted C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸COR⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR^B, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;
R⁸, R⁹and R¹⁰each independently of one another denote hydrogen or a group selected from among optionally substituted C_1-8-alkyl, C_2-8-alkenyl, C_2-8-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, while the substituent(s) may be identical or different and are selected from among halogen, —NH₂, —OH and pseudohalogen; optionally in the form of the tautomers, racemates, enantiomers, diastereomers and mixtures thereof, and optionally the pharmacologically acceptable acid addition salts thereof, and
at least one other cytostatic or cytotoxic active substance, optionally in the form of the tautomers, racemates, enantiomers, diastereomers and mixtures thereof, and optionally the pharmacologically acceptable acid addition salts thereof.

DEFINITIONS

As used herein, the following definitions apply, unless stated otherwise.
By alkyl substituents are meant in each case saturated, straight-chain or branched aliphatic hydrocarbon groups (alkyl group).
The alkenyl substituents are in each case straight-chain or branched, unsaturated alkyl groups which have at least one double bond.
By alkynyl substituents are meant in each case straight-chain or branched, unsaturated alkyl groups which have at least one triple bond.
Haloalkyl refers to alkyl groups wherein one or more hydrogen atoms are replaced by halogen atoms. Haloalkyl includes both saturated alkyl groups and unsaturated alkenyl and alkynyl groups, such as for example —CF₃, —CHF₂, —CH₂F, —CF₂CF₃, —CHFCF₃, —CH₂CF₃, —CF₂CH₃, —CHFCH₃, —CF₂CF₂CF₃, —CF₂CH₂CH₃, —CHFCH₂CH₃and —CHFCH₂CF₃.
Halogen relates to fluorine, chlorine, bromine and/or iodine atoms.
By pseudohalogen are meant the following groups: —OCN, —SCN, —CF₃and —CN.
By cycloalkyl is meant a mono- or bicyclic ring, while the ring system may be a saturated ring or an unsaturated, non-aromatic ring, which may optionally also contain double bonds, such as for example cyclopropyl, cyclopropenyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, norbornyl, norbornenyl, spiro[5.5]undecane, spiro[5.4]decane and spiro[4.4]nonane.
Aryl relates to monocyclic or bicyclic rings with 6-12 carbon atoms such as for example phenyl and naphthyl.
By heteroaryl are meant mono- or bicyclic rings which contain instead of one or more carbon atoms one or more identical or different heteroatoms, such as e.g. nitrogen, sulphur or oxygen atoms. Examples include furyl, thienyl, pyrrolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl, pyridyl, pyrimidyl, pyridazinyl, pyrazinyl and triazinyl. Examples of bicyclic heteroaryl groups are indolyl, isoindolyl, benzofuranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolyl, indazolyl, isoquinolinyl, quinolinyl, quinoxalinyl, cinnolinyl, phthalazinyl, quinazolinyl and benzotriazinyl, indolizinyl, oxazolopyridinyl, imidazopyridinyl, naphthyridinyl, indolinyl, isochromanyl, chromanyl, tetrahydroisoquinolinyl, isoindolinyl, isobenzotetrahydrofuranyl, isobenzotetrahydrothienyl, isobenzothienyl, benzoxazolyl, pyridopyridinyl, benzotetrahydrofuranyl, benzotetrahydrothienyl, purinyl, benzodioxolyl, triazinyl, phenoxazinyl, phenothiazinyl, pteridinyl, benzothiazolyl, imidazopyridinyl, imidazothiazolyl, dihydrobenzisoxazinyl, benzisoxazinyl, benzoxazinyl, dihydrobenzisothiazinyl, benzopyranyl, benzothiopyranyl, cumarinyl, isocumarinyl, chromonyl, chromanonyl, pyridinyl-N-oxid, tetrahydroquinolinyl, dihydroquinolinyl, dihydroquinolinonyl, dihydroisoquinolinonyl, dihydrocumarinyl, dihydroisocumarinyl, isoindolinonyl, benzodioxanyl, benzoxazolinonyl, pyrrolyl-N-oxide, pyrimidinyl-N-oxide, pyridazinyl-N-oxide, pyrazinyl-N-oxide, quinolinyl-N-oxide, indolyl-N-oxide, indolinyl-N-oxide, isoquinolyl-N-oxide, quinazolinyl-N-oxide, quinoxalinyl-N-oxide, phthalazinyl-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, benzothiopyranyl-S-oxide and benzothiopyranyl-S,S-dioxide.
Heterocyclyl relates to saturated or unsaturated, non-aromatic mono-, bicyclic or bridged bicyclic rings comprising 5-12 carbon atoms, which carry heteroatoms, such as nitrogen, oxygen or sulphur, instead of one or more carbon atoms. Examples of such heterocylyl groups are tetrahydrofuranyl, pyrrolidinyl, pyrrolinyl, imidazolidinyl, imidazolinyl, pyrazolidinyl, pyrazolinyl, piperidyl, piperazinyl, indolinyl, isoindoliny, morpholinyl, thiomorpholinyl, homomorpholinyl, homopiperidyl, homopiperazinyl, thiomorpholinyl-S-oxide, thiomorpholinyl-S,S-dioxide, tetrahydropyranyl, piperidinyl, tetrahydrothienyl, homopiperidinyl, homothiomorpholinyl-S,S-dioxide, oxazolidinonyl, dihydropyrazolyl, dihydropyrrolyl, dihydropyrazinyl, dihydropyridinyl, dihydropyrimidinyl, dihydrofuryl, dihydropyranyl, tetrahydrothienyl-5-oxide, tetrahydrothienyl-S,S-dioxide, homothiomorpholinyl-5-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, 2,6-diaza-bicyclo[3.2.2]nonane, 2,7-diaza-spiro[3.5]nonane, 2,7-diaza-spiro[4.4]nonane, 2,8-diaza-spiro[4.5]decane, 3,9-diaza-spiro[5.5]undecane.
The Examples that follow illustrate the present invention without restricting its scope:

Preparation of the Compounds According to the Invention:

The compounds according to the invention may be prepared according to methods of synthesis A to C described hereinafter, wherein the substituents of general formulae (I to XVI) have the meanings given hereinbefore.

Method A

Step 1A

The intermediate compound III is prepared by substitution of a leaving group LG, for example halogen, SCN or methoxy, preferably chlorine, in a heteroaromatic system I by a nucleophile II.
1 equivalent of compound I and 1 to 1.5 equivalents of compound II are stirred in a solvent, for example 1,4-dioxane, tetrahydrofuran, ethanol, isopropanal, N,N-dimethylformamide or N,N-dimethylacetamide. At a temperature of 15 to 25° C., 2 to 2.5 equivalents of a base, for example potassium carbonate, sodium carbonate, caesium carbonate, N-ethyl-N,N-diisopropylamine or triethylamine, are added. The reaction mixture is stirred for 6 to 72 h at a temperature of 20 to 100° C. Then the solvent is distilled off and the residue is combined with water which has been adjusted to a pH of between 1-4 with an inorganic acid, for example hydrochloric acid or sulphuric acid. This mixture is extracted two to three times with an organic solvent, for example diethyl ether, ethyl acetate or dichloromethane. The combined organic extracts are dried and the solvent is distilled off. The residue is purified by chromatography.

Step 2A

The end compound V is prepared by substitution of a leaving group LG, for example halogen, SCN or methoxy, preferably chlorine, in a heteroaromatic system III by a nucleophile IV.
1 equivalent of the compound III and 1 to 3 equivalents of the compound IV are stirred in a solvent, for example 1,4-dioxane, N,N-dimethylformamide, N,N-dimethylacetamide or N-methyl-2-pyrrolidinone. At a temperature of 15 to 40° C., 1 to 2 equivalents of an inorganic acid, for example sulphuric acid or hydrochloric acid, are added. The reaction mixture is stirred for another 12 to 72 h at a temperature of 20 to 100° C. Then the solvent is distilled off and the residue is purified by chromatography.

Method B

Step 1B

The intermediate compound VII is prepared by substitution of a leaving group LG, for example halogen, SCN, methoxy, preferably chlorine, in a heteroaromatic system I by a nucleophile VI.
1 equivalent of the compound I and 1 to 1.5 equivalents of the compound VI are stirred in a solvent, for example 1,4-dioxane, tetrahydrofuran, ethanol, isopropanol, N,N-dimethylformamide or N,N-dimethylacetamide.
At a temperature of 15 to 25° C., 2 to 2.5 equivalents of a base, for example potassium carbonate, sodium carbonate, caesium carbonate, potassium hydrogen phosphate, N-ethyl-N,N-diisopropylamine or triethylamine are added. The reaction mixture is stirred for 6 to 72 h more at a temperature of 20 to 120° C. The reaction mixture is combined with water, which has been adjusted to a pH of 8 to 9 with an inorganic base, for example sodium hydrogen carbonate or potassium carbonate. This mixture is extracted two to three times with an organic solvent, for example diethyl ether or ethyl acetate.
The combined organic extracts are dried and the solvent is distilled off. The residue is purified by chromatography or repeated crystallisation.

Step 2B

The intermediate compound VIII is prepared by substituting a leaving group LG, for example halogen, SCN, methoxy, preferably chlorine, in a heteroaromatic system VII by a nucleophile IV.
1 equivalent of the compound VII and 1 to 1.5 equivalents of the compound IV are stirred in a solvent, for example 1,4-dioxane, N,N-dimethylformamide, N,N-dimethylacetamide or N-methyl-2-pyrrolidinone. At a temperature of 15 to 40° C., 0.2 to 1 equivalent of an acid, for example sulphuric acid or hydrochloric acid, is added. The reaction mixture is stirred for another 12 to 72 h at a temperature of 20 to 100° C. The reaction mixture is stirred into water and the resulting precipitate is filtered off and dried. The precipitate may be purified by chromatography or crystallisation or used as the crude product in the next step.

Step 3B

Compounds VIII whose group R⁷denotes hydrogen may be used directly for preparing the end compounds X, while a compound VIII is reacted with a compound IX.
Compounds VIII whose group R⁷does not denote hydrogen are converted beforehand by hydrolysis or similar methods known to the skilled man into the compounds wherein the group R⁷denotes hydrogen.
1 equivalent of the compound VIII, 1 to 1.5 equivalents of the compound I× and 1 to 3 equivalents of a base, for example triethylamine or ethyldiisopropylamine, are stirred in a solvent, for example 1,4-dioxane, N,N-dimethylformamide, N,N-dimethylacetamide or N-methyl-2-pyrrolidinone. At a temperature of 15 to 25° C., 1 to 1.5 equivalents of a coupling reagent, for example N,N-dicyclohexylcarbodiimide, N,N-diisopropyl-carbodiimide, O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-tetrafluoroborate or 1-(3-N,N-dimethylaminopropyl)-3-ethylcarbodiimide are added. The reaction mixture is stirred for another 4 to 24 h at a temperature of 15 to 25° C. Then the solvent is distilled off and the residue is purified by chromatography.

Method C

Step 1C

The intermediate compound XI is prepared by substituting a leaving group LG, for example halogen, SCN, methoxy, preferably chlorine, at a heteroaromatic system I with a nucleophilic group IV.
1 equivalent of the compound I and 1 to 3 equivalents of a base, for example triethylamine or ethyldiisopropylamine, are stirred in a solvent, for example 1,4-dioxane, tetrahydrofuran, N,N-dimethylformamide or N,N-dimethylacetamide. At a temperature of −60 to 0° C., 0.8 to 1.5 equivalents of a compound IV are added. The reaction mixture is stirred for 6 to 72 h at a temperature of 15 to 75° C. Then the solvent is distilled off and the residue is purified by chromatography.

Step 2C

The end compound V is prepared by substitution of a leaving group LG, for example halogen, SCN, methoxy, preferably chlorine, at a heteroaromatic system XI by a nucleophile II.
1 equivalent of the compound XI and 1 to 1.5 equivalents of the compound II are stirred in a solvent, for example 1,4-dioxane, N,N-dimethyl-formamide, N,N-dimethylacetamide or N-methyl-2-pyrrolidinone. At a temperature of 15 to 40° C. 1 to 2 equivalents of an acid, for example sulphuric acid or hydrochloric acid, are added. The reaction mixture is stirred for another 6 to 72 h at a temperature of 20 to 100° C. Then the solvent is distilled off and the residue is purified by chromatography.

Chromatography:

For medium pressure chromatography (MPLC) silica gel made by Millipore (name: Granula Silica Si-60A 35-70 μm) or C-18 RP-silica gel made by Macherey Nagel (name: Polygoprep 100-50 C18) is used.
For high pressure chromatography columns made by Waters (name: XTerra Prep. MS C18, 5 μM, 30*100 mm or Symmetrie C18, 5 μm, 19*100) are used.

Nuclear Magnetic Resonance (NMR) Spectroscopy:

The measurement is carried out in deuterised dimethylsulphoxide-d6. If other solvents are used they are explicitly mentioned in the Examples or in the methods. The measurements are given on a delta scale in ppm. Tetramethylsilane is taken as the standard. The measurements are carried out on an Avance 400 (400 MHz NMR spectrometer) made by Messrs Bruker Biospin GmbH.
The NMR spectra are given purely in a descriptive capacity. Basically, only the visible molecular signals are listed. If for example molecular signals are partly or completely masked by foreign signals such as for example water signals, DMSO signals or CDCl₃signals they are not mentioned.

Mass Spectroscopy/UV Spectrometer:

These data are generated using an HPLC-MS apparatus (high performance liquid chromatography with mass detector) made by Agilent.
The apparatus is constructed so that a diode array detector (G1315B made by Agilent) and a mass detector (1100 LS-MSD SL; G1946D; Agilent) are connected in series downstream of the chromatography apparatus (column: Zorbax SB-C8, 3.5 μm, 2.1*50, Messrs. Agilent). The apparatus is operated with a flow of 0.6 ml/min For a separation process a gradient is run through within 3.5 min (start of gradient: 95% water and 5% acetonitrile; end of gradient: 5% water and 95% acetonitrile; in each case 0.1% formic acid is added to the two solvents).

Method 1

2-(2-methoxy-4-propylcarbamoyl-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

5 g (21.9 mmol) 2,4-dichloro-5-trifluoromethyl-pyrimidine are dissolved in 50 ml 1,4-dioxane and combined with 5.5 g (21.9 mmol) 4-amino-3-methoxybenzoic acid-propylamide hydrochloride (Zhuangyu Zhang, et al. 1989, J Pharml Sci. 78(10):829-32). 7.5 ml (43.8 mmol) ethyldiisopropylamine are added to this reaction mixture and the mixture is stirred for 2 days at ambient temperature. Then the reaction mixture is diluted with 250 ml of ethyl acetate and washed first with 300 ml aqueous 10% KHSO₄solution, then with 300 ml saturated aqueous NaCl solution. The organic phase is dried with MgSO₄and the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant is a mixture of cyclohexane:ethyl acetate (75:25).
Yield: 2.30 g (5.9 mmol; 27%)
¹H-NMR: 0.91 (t, 3H), 1.50-1.61 (m, 2H), 3.20-3.28 (m, 2H), 3.87 (s, 3H), 7.46-7.51 (m, 1H), 7.52-7.56 (m, 1H), 7.70-7.75 (m, 1H), 8.44 (t, 1H), 8.75 (s, 1H), 9.73 (s, 1H)

Method 2

7-amino-2,3-dihydro-isoindol-1-one

a) 7-nitro-2,3-dihydro-isoindol-1-one

1.5 g (5.473 mmol) methyl 2-bromomethyl-6-nitro-benzoate are dissolved in 20 ml N,N-dimethylformamide and combined with 15 ml of methanolic ammonia (7 mmol/ml). After 20 h at 25° C. the mixture is diluted with 100 ml of ethyl acetate and extracted 3 times with saturated sodium hydrogen carbonate solution. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo.
Yield: 960 mg (5.389 mmol, 99%)
MS-ESI+: m/z=179 [M+H]⁺

b) 7-amino-2,3-dihydro-isoindol-1-one

960 mg (5.389 mmol) 7-nitro-2,3-dihydro-isoindol-1-one are dissolved in 100 ml of tetrahydrofuran and combined with 100 mg palladium on charcoal. Then the mixture is stirred for 20 h at 25° C. and 4 bar hydrogen pressure (H₂pressure). The catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 734 mg (4.958 mmol, 92%)
MS-ESI+: m/z=149 [M+H]⁺
The following 7-amino-2,3-dihydro-isoindol-1-one derivatives are prepared analogously to this method. A corresponding amine is used instead of ammonia:


		MS (ESI)
		(M + H)⁺

		163

		177

		191

		231

		219

		233

		207

		234

		274

		195

		213

		231

		209

		245

		188

		187

		206

		233

		233

		202

		206

		191

		205

		227

		223

		193

		225

		243

		221

		255

		192

		255

		178

		192

		211/213

		247

		247

		261

		261

		261

		261

		223

		223

		221

		247

		246

		235

		224

		222

Method 3

Ethyl (4-amino-3-oxo-1,3-dihydro-isobenzofuran-1-yl)-acetate

a) ethyl (4-amino-3-oxo-3H-isobenzofuran-1-ylidene)-acetate

500 mg (3.1 mmol) 4-amino-isobenzofuran-1,3-dione and 1.13 g (3.1 mmol) (ethoxy-carbonylmethylene)-triphenylphosphorane are dissolved in 5 ml of tetrahydrofuran (THF) and refluxed for 3 h. Then the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is a mixture of cyclohexane:ethyl acetate (75:25).
Yield: 221 mg (0.95 mmol, 31%)
MS-ESI+: m/z=234 [M+H]⁺

b) ethyl (4-amino-3-oxo-1,3-dihydro-isobenzofuran-1-yl)-acetate

120 mg (0.51 mmol) ethyl (4-amino-3-oxo-3H-isobenzofuran-1-ylidene)-acetate are dissolved in 50 ml of methanol and combined with 50 mg palladium on activated charcoal (10% Pd). The reaction mixture is hydrogenated for 3 h at 2 bar H₂pressure and 25° C. Then the catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 116 mg (0.49 mmol, 97%)
MS (ESI): m/z=236 (M+H)⁺
¹H-NMR: 1.17 (t, 3H), 2.68-2.78 (m, 1H), 3.08-3.16 (m, 1H), 4.10 (q, 2H), 5.67-5.74 (m, 1H), 6.28 (bs, 2H), 6.61-6.70 (m, 2H), 7.30-7.38 (m, 1H)

Method 4

5-amino-3H-quinazolin-4-one

a) 2,6-diaminobenzamide

5 g (25.373 mmol) 2,6-dinitro-benzonitrile is combined with 20 ml of an aqueous 80% sulphuric acid and stiffed for 2 h at 80° C. The reaction mixture is combined with 100 ml of tetrahydrofuran and neutralised with 10% aqueous sodium hydroxide solution. The organic phase is separated off, combined with another 100 ml of tetrahydrofuran and 200 mg palladium on charcoal and stirred for 20 h at 8 bar H₂pressure and 25° C. The solids are filtered off. The filtrate is combined with 300 ml of ethyl acetate and extracted with saturated potassium hydrogen carbonate solution. The organic phase is separated off, dried and the solvent is eliminated in vacuo. The residue is purified by chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 7% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution are added.
Yield: 900 mg (5.958 mmol; 23%)
MS (ESI): 152 (M+H)⁺

b) 5-amino-3H-quinazolin-4-one

900 mg (5.958 mmol) 2,6-diaminobenzamide are dissolved in 3.6 ml N,N-dimethylacetamide and combined with 6.3 ml (57.01 mmol) trimethylorthoformate and 792 μl (8.865 mmol) 98% sulphuric acid. After 16 h at 25° C. the reaction mixture is taken up with 20 ml of methanol and the solvent is eliminated in vacuo. The residue is again taken up in 20 ml of methanol, neutralised with concentrated ammonia. The solvent is eliminated in vacuo and the residue purified by chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 7% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution are added.
Yield: 782 mg (4.852 mmol; 81%)
MS (ESI): 162 (M+H)⁺

Method 5

9-amino-2,3,4,5-tetrahydro-2-benzazepin-1-one

500 mg (1.825 mmol) 2-bromomethyl-6-nitro-methylbenzoate are heated to 100° C. in 2 ml trimethyl phosphate for 5 h. 2-(dimethylphosphonomethyl)-6-nitromethylbenzoate is obtained by evaporation under a high vacuum and used further directly. The crude product is dissolved in 24 ml of tetrahydrofuran at −70° C. under N₂, 2.7 ml (2.7 mmol) of a 1 M lithium hexamethyldisilazide solution in tetrahydrofuran is added dropwise and then 430 mg (2.70 mmol) tert.-butyl-N-(2-oxoethyl)-carbamate in 5 ml of tetrahydrofuran are added. The reaction mixture is slowly heated to ambient temperature, combined with 5 ml of 1 M HCl and extracted with ethyl acetate. The combined organic phases are concentrated by evaporation and, by chromatography on silica gel with a mixture of cyclohexane-ethyl acetate in the ratio 95:5 to 75:25, 338 mg (1.006 mmol, 55%) of the E-/Z mixture of 2-(3-tert.-butoxycarbonylamino-prop-1-en-1-yl)-6-nitro-methylbenzoate are obtained. This E-/Z-mixture is treated for 12 h with 10 ml of a saturated methanolic potassium hydroxide solution. After acidification with aqueous 1 M HCl and extraction with ethyl acetate 302 mg (0.938 mmol, 93%) of the E-/Z mixture of 2-(3-tert.-butoxycarbonylamino-prop-1-en-1-yl)-6-nitro-methylbenzoic acid are obtained. To this are added 20 mg Raney nickel in 100 ml of methanol and the mixture is hydrogenated at 5 bar H₂pressure. The catalyst is filtered off, the filtrate concentrated by evaporation and stirred overnight with a 1:1 mixture of trifluoroacetic acid and dichloromethane at ambient temperature. After elimination of the solvent 133 mg (0.686 mmol, 73%) 2-amino-6-(3-amino-propyl)-benzoic acid are obtained. The further reaction is carried out by dissolving in 10 ml THF and 10 ml DCM with the addition of 300 mg (1.570 mmol) N-(3-dimethylaminopropyl)-N⁴-ethylcarbodiimide hydrochloride and 134 μl (0.830 mmol) N,N-diisopropyl-ethylamine and 48 h stirring at ambient temperature. The solvent is eliminated in vacuo and the crude product is purified by chromatography with C18-RP silica gel and an eluant mixture of acetonitrile and water in the ratio 5:95 to 95:5, to which 0.1% formic acid has been added.
Yield: 28 mg (0.160 mmol, 23%)
MS (ESI): m/z=177 (M+H)⁺

Method 6

4-amino-1-methyl-1,2-dihydro-indazol-3-one

a) 4-nitro-1,2-dihydro-indazol-3-one

5 g (27.5 mmol) 2-amino-6-nitro-benzoic acid are combined with 22.2 ml (225.3 mmol) concentrated HCl and 45 ml (30.0 mmol) 5% aqueous sodium nitrite solution and stirred for 1 h at ambient temperature. Then the suspension is diluted with 150 ml dist. H₂O and added dropwise to 350 ml destilliertes water which has been saturated with sulphur dioxide. Sulphur dioxide is piped through the reaction mixture for a further 30 min. Then the reaction mixture is refluxed for 30 min and then left to cool slowly to 20° C. The resulting precipitate is filtered off.
Yield: 1.7 g (9.5 mmol, 35%)
MS (ESI): m/z=180 (M+H)⁺

b) 1-methyl-4-nitro-1,2-dihydro-indazol-3-one

306 mg (1.7 mmol) 4-nitro-1,2-dihydro-indazol-3-one are dissolved in 1 ml N,N-dimethyl-acetamide, combined with 150 μl (2.4 mmol) methyl iodide and 500 μl (2.32 mmol) of N-ethyldiisopropylamide and stiffed for 2 h at ambient temperature. Then the reaction mixture is combined with 40 ml of a 1 N aqueous hydrochloric acid and extracted twice with 50 ml dichloromethane. Then the organic phase is dried with MgSO₄, the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and 5% water and 95% acetonitrileat the finishing point.
Yield: 144 mg (0.7 mmol, 44%)
MS (ESI): m/z=194 (M+H)⁺
¹H-NMR: 3.90 (s, 3H), 7.47-7.52 (m, 1H), 7.68-7.73 (m, 1H), 7.88-7.93 (m, 1H), 10.53 (s, 1H)

c) 4-amino-1-methyl-1,2-dihydro-indazol-3-one

140 mg (0.7 mmol) 1-methyl-4-nitro-1,2-dihydro-indazol-3-one are suspended in 6 ml of ethanol and combined with 600 mg (4.4 eq, 2.9 mmol) sodium dithionite, dissolved in 2 ml distilled water, and stirred for 15 min at 25° C. Then the reaction mixture is combined with distilled water and extracted twice with ethyl acetate. Then the organic phase is dried with MgSO₄and the solvent is eliminated in vacuo.
Yield: 33 mg (0.2 mmol, 28%)
MS (ESI): m/z=164 (M+H)⁺
4-amino-1,2-dihydro-indazol-3-one and the following compounds are prepared analogously to this method.


		MS (ESI)
		(M + H)⁺

		178

		194

		178

Method 7

8-amino-4-methyl-3,4-dihydro-2H-isoquinolin-1-one

a) methyl 2-(cyanomethyl-2-methyl)-6-nitro-benzoate

400 mg (1.8 mmol) methyl 2-cyanomethyl-6-nitro-benzoate are dissolved in 13 ml THF, combined with 114 μl (1.8 mmol) methyl iodide and the mixture is cooled to −20° C. under a nitrogen atmosphere. Then at this temperature 250 mg (2.2 mmol) potassium-tert-butoxide are added. After 1 h the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and 5% water and 95% acetonitrile at the finishing point.
Yield: 289 mg (1.2 mmol, 68%)
MS (ESI): 233 (M−H)⁻

b) 8-amino-4-methyl-3,4-dihydro-2H-isoquinolin-1-one

400 mg (1.8 mmol) methyl 2-(cyanomethyl-2-methyl)-6-nitro-benzoate are dissolved in 13 ml of methanol and combined with 50 mg Raney nickel. The reaction mixture is hydrogenated for 16 h at 4 bar H₂pressure and 25° C. Then the catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 170 mg (0.8 mmol, 46%)
MS (ESI): 177 (M+H)⁺
8-amino-3,4-dihydro-2H-isoquinolin-1-one and 8-amino-4,4-dimethyl-3,4-dihydro-2H-isoquinolin-1-one and the following compounds are prepared analogously to this method.


		MS (ESI)
		(M + H)⁺

		221

		253

		205

Method 8

7-amino-indan-1-one

a) indan-4-ylamine

24 ml (349 mmol) 65% nitric acid are cooled to 0-5° C. 28 ml (518.5 mmol) of concentrated sulphuric acid are slowly added dropwise while cooling with ice. This solution is cooled to 5° C. and slowly added dropwise to 30 ml (232 mmol) indane cooled to 0-5° C., with vigorous stirring and further cooling with ice. The reaction mixture is stirred for 30 min at 0-5° C., and then heated to 25° C. for 1 h with stirring. Then the solution is added dropwise to 150 ml ice/water and stirred for 30 min The aqueous phase is extracted three times with 200 ml diethyl ether. The combined organic phases are washed twice with 200 ml saturated sodium hydrogen carbonate solution and once with 150 ml distilled water. Then the organic phase is dried with MgSO₄and the solvent is eliminated in vacuo. The crude product is dissolved in 250 ml of methanol and combined with 4.5 g Raney nickel. The reaction mixture is hydrogenated for 16 h at 3 bar H₂pressure and 25° C. Then the catalyst is filtered off and the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is a mixture of cyclohexane:ethyl acetate (75:25).
Yield: 3.81 g (28.6 mmol, 12%)
MS (ESI): 134 (M+H)⁺
¹H-NMR: 1.90-2.00 (m, 2H), 2.61 (t, 2H), 2.76 (t, 2H), 4.73 (s, 2H), 6.33-6.38 (m, 1H), 6.39-6.45 (m, 1H), 6.76-6.83 (m, 1H)

b) N-indan-4-yl-acetamide

226 mg (1.7 mmol) indan-4-ylamine are combined with 5 ml acetic anhydride. The suspension is stirred for 16 h at 70° C. The resulting solution is stirred into 40 ml distilled water, adjusted to pH 7 with sodium carbonate and extracted three times with 30 ml of ethyl acetate. Then the organic phase is dried with MgSO₄, the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is a mixture of cyclohexane:ethyl acetate (70:30).
Yield: 152 mg (0.9 mmol, 51%)
MS (ESI): 176 (M+H)⁺
¹H-NMR: 1.93-2.03 (m, 2H), 2.04 (s, 3H), 2.79 (t, 2H), 2.86 (t, 2H), 6.94-7.01 (m, 1H), 7.02-7.10 (m, 1H), 7.36-7.44 (m, 1H), 9.25 (s, 1H)

c) N-(3-oxo-indan-4-yl)-acetamide

147 mg (0.84 mmol) N-indan-4-yl-acetamide are dissolved in 10 ml acetone and combined with 770 μl of a 15% aqueous magnesium sulphate solution. The solution is cooled to 0° C. and 397 mg (2.490 mmol) potassium permanganate are added batchwise. After 2 h the mixture is diluted with 50 ml of water, and extracted three times with 20 ml chloroform. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is a mixture of cyclohexane:ethyl acetate (85:15).
Yield: 95 mg (0.500 mmol, 60%)
MS (ESI): 190 (M+H)⁺

d) 7-amino-indan-1-on

500 mg (2.6 mmol) N-(3-oxo-indan-4-yl)-acetamide are dissolved in 5 ml of ethanol, combined with 5 ml 18% hydrochloric acid and stirred for 3 h at 70° C. Then the reaction mixture is stirred into 100 ml distilled water, adjusted to pH 7 with sodium carbonate and extracted three times with 30 ml of ethyl acetate. Then the organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo.
Yield: 388 mg (2.6 mmol, 100%)
8-amino-3,4-dihydro-2H-naphthalen-1-one is prepared analogously to this method. 1,2,3,4-tetrahydronaphthalene is used as starting material instead of indane.

Method 9

N-(7-amino-1-oxo-1,3-dihydro-isoindol-2-yl)-acetamide

a) 2-benzyloxy-N-(7-nitro-1-oxo-1,3-dihydro-isoindol-2-yl)-acetamide

870 mg (4.5 mmol) 2-amino-7-nitro-2,3-dihydro-isoindol-1-one (prepared analogously to method 2) are dissolved in 82 ml dichloromethane and 64 ml THF. The solution is combined with 2.8 ml (3.3 eq, 20 mmol) benzyloxyacetyl chloride, 4.8 ml (28.0 mmol) N-ethyldiisopropyl-amine and 10 mg N,N-dimethylaminopyridine and stirred for 3 h at 25° C. Then the reaction mixture is combined with 100 ml aqueous 0.1 N hydrochloric acid and extracted three times with 50 ml of ethyl acetate. The organic phase is dried with magnesium sulphate, the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is a mixture of dichloromethane:methanol (95:5).
Yield: 910 mg (2.7 mmol, 59%)

b) N-(7-amino-1-oxo-1,3-dihydro-isoindol-2-yl)-acetamide

790 mg (2.3 mmol) 2-benzyloxy-N-(7-nitro-1-oxo-1,3-dihydro-isoindol-2-yl)-acetamide are dissolved in 100 ml of methanol and combined with 80 mg palladium hydroxide. The reaction mixture is hydrogenated for 48 h at 4 bar H₂pressure and 25° C. Then the catalyst is filtered off and the solvent is eliminated in vacuo. The crude product is purified by chromatography. The carrier used is silica gel and the eluant used is a mixture of dichloromethane:methanol (90:10).
Yield: 210 mg (0.1 mmol, 41%)
MS (ESI): 222 (M+H)⁺

Method 10

6-amino-2-ethyl-3,4-dihydro-2H-benzo[f][1,4]oxazepin-5-one

a) 2-amino-6-(1-aminomethyl-propoxy)-benzonitrile

2.01 g (22 mmol) 1-amino-2-butanol are dissolved in 6.5 ml 1,4-dioxane, combined with 880 mg (7.8 mmol) sodium hydride and stirred for 30 min at ambient temperature. 2 g (14.7 mmol) of 2-amino-6-fluorobenzonitrile are added to this reaction mixture and it is stirred for 24 h at 50° C. Then the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 5% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution has been added.
Yield: 1.15 g (5.6 mmol, 38%)
MS (ESI): 206 (M+H)⁺

b) 2-amino-6-(1-aminomethyl-propoxy)-benzoic acid

1.15 g (5.6 mmol) 2-amino-6-(1-aminomethyl-propoxy)-benzonitrile are dissolved in 10 ml 20% ethanolic KOH and stirred for 24 h at 80° C. Then the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 12% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
Yield: 262 mg (1.2 mmol, 21%)
MS (ESI): 225 (M+H)⁺

c) 6-amino-2-ethyl-3,4-dihydro-2H-benzo[f][1,4]oxazepin-5-one

262 mg (1.2 mmol) 2-amino-6-(1-aminomethyl-propoxy)-benzoic acid are dissolved in 26 ml THF, combined with 680 mg (3.5 mmol) 1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide hydrochloride and 0.6 ml (3.5 mmol) diisopropyl-ethylamine and stirred for 3 h at 50° C. Then the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 4% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
Yield: 50 mg (0.2 mmol, 21%)
MS (ESI): 207 (M+H)⁺
The following compounds are prepared analogously to this method. 1-amino-2-butanol was replaced by a corresponding aminoalcohol or by a corresponding 1,2-diaminoethylene.


		MS (ESI)
		(M + H)⁺

		207

		193

		235

		219

		233

		207

		207

		193

		221

		299

		219

		209

		269

		251

		179

		221

		206

		235

		227

		219

		207

		269

		225

		253

		241

		233

Method 11

6-amino-3-benzyl-3,4-dihydro-1H-benzo[e][1,4]diazepine-2,5-dione

a) methyl 2-(2-amino-6-nitro-benzoylamino)-3-phenyl-propionate

1.18 g (6.5 mmol) 2-amino-6-nitrobenzoic acid, 1.0 g (4.6 mmol) D,L-phenylalanine-methylester hydrochloride, 4.05 ml (23.2 mmol) N-ethyldiisopropylamine are combined with 2.5 ml of tetrahydrofuran. 1.71 g (5.1 mmol) O-(benzotriazol-1-yl)-N,N,N′N′-tetramethyluronium-tetrafluoroborate are added to this reaction mixture and it is heated for 12 h to 50° C. Then the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is a mixture of cyclohexane:ethyl acetate (50:50).
Yield: 1.04 g (3.03 mmol, 65%)
MS (ESI): 344 (M+H)⁺

b) 2-(2-amino-6-nitro-benzoylamino)-3-phenyl-propionic acid

1.04 g (3.03 mmol) methyl 2-(2-amino-6-nitro-benzoylamino)-3-phenyl-propionate are dissolved in 3 ml 20% ethanolic KOH and stirred for 1.5 h at 50° C. Then the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 15% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution has been added.
Yield: 636 mg (1.9 mmol, 64%)
MS (ESI): 329 (M+H)⁺
¹H-NMR: 2.86-2.94 (m, 1H), 3.17 (s, 1H), 3.22-3.29 (m, 1H), 4.30-4.38 (m, 1H),
6.63 (s, 2H), 6.89-6.96 (m, 1H), 6.97-7.02 (m, 1H), 7.12-7.21 (m, 2H), 7.21-7.27 (m, 2H), 7.28-7.35 (m, 2H), 8.33-8.43 (m, 1H)

c) 2-(2,6-diamino-benzoylamino)-3-phenyl-propionic acid

410 mg (1.25 mmol) 2-(2-amino-6-nitro-benzoylamino)-3-phenyl-propionic acid are dissolved in 50 ml of methanol and combined with 40 mg palladium on charcoal (10% Pd). The reaction mixture is hydrogenated for 9 h at 5 bar H₂pressure and 25° C. Then the catalyst is filtered off, the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 5% water and 95% acetonitrile at the finishing point.
Yield: 88 mg (0.29 mmol, 24%)
MS (ESI): 300 (M+H)⁺

d) 6-amino-3-benzyl-3,4-dihydro-1H-benzo[e][1,4]diazepine-2,5-dione

88 mg (0.3 mmol) 2-(2,6-diamino-benzoylamino)-3-phenyl-propionic acid are dissolved in 2 ml THF, combined with 143 mg (0.9 mmol) 1-(3-dimethyl-aminopropyl)-3-ethylcarbodiimide hydrochloride and 103 μl (0.6 mmol) diisopropyl-ethylamine and stirred for 17 h at 50° C. Then the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 5% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
Yield: 22 mg (0.08 mmol, 27%)
MS (ESI): 282 (M+H)⁺
The following compounds are prepared analogously to to method 11.


		MS (ESI)
		(M + H)⁺

		192

		206

		206

		218

		220

		220

		232

		232

		234

		234

		234

		246

		246

		246

		248

		248

		248

		250

		265

		265

		268

		277

		278

		278

		282

		283

		283

		288

		296

		192

		298

		298

		300

		300

		300

		307

		316/318

		321

		321

		346

Method 12

2-(4-carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

7.36 g (44 mmol) 4-amino-3-methoxybenzoic acid are suspended in 80 ml of an aqueous phosphate buffer solution (pH 6.3) and combined with 9.5 g (44 mmol) 2,4-dichloro-5-trifluoro-methyl-pyrimidine, which is dissolved in 240 ml 1,4-dioxane. After 4 h at 100° C. the reaction mixture is crystallised at 0° C. The precipitate is filtered off, the filtrate is combined with 150 ml of ethyl acetate and washed twice with 200 ml of a saturated aqueous sodium hydrogen carbonate solution. The organic phase is dried with MgSO₄and the solvent is eliminated in vacuo. The crude product is suspended in 10 ml n-hexane and refluxed. The precipitate is filtered off, suspended in 48 ml of a saturated aqueous sodium hydrogen carbonate solution and heated to 65° C. for 1 h. Then the solution is crystallised at 0° C. The precipitate is filtered off, the filtrate is acidified with 1 N aqueous hydrochloric acid and combined with 100 ml of ethyl acetate. The organic phase is separated off, dried with magnesium sulphate and the solvent is eliminated in vacuo. The residue is recrystallised from ethyl acetate.
Yield: 330 mg (0.95 mmol, 2%)
MS (ESI): 348 (M+H)⁺
¹H-NMR: 1.55 (s, 1H), 4.01 (s, 3H), 7.61-7.64 (m, 1H), 7.79-7.85 (m, 1H), 8.34
(s, 1H), 8.59-8.63 (m, 1H), 8.66 (s, 1H)

Method 13

4-(4-amino-cyclohexyl)-morpholine

a) dibenzyl-(4-morpholino-4-yl-cyclohexyl)-amine

3.9 g (30 mmol) 4-dibenzylamino-cyclohexanone are dissolved in 100 ml dichloromethane and stirred with 3.9 g (45 mmol) morpholine and 9.5 g (45 mmol) sodium triacetoxyborohydride for 12 h at ambient temperature. Then water and potassium carbonate are added, the organic phase is separated off, dried and the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is ethyl acetate, to which 10% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added. The suitable fractions are evaporated down in vacuo.
Yield: 6.6 g (18 mmol, 60%) cis-isomer
2 g (5.4 mmol, 18%) trans-isomer.

b) trans-4-morpholino-4-yl-cyclohexylamine

7.2 g (16.4 mmol) trans-dibenzyl-4-morpholino-cyclohexylamine are dissolved in 100 ml of methanol and hydrogenated on 1.4 g palladium on charcoal (10% Pd) at 30-50° C. The solvent is eliminated in vacuo and the residue is crystallised from ethanol and concentrated hydrochloric acid.
Yield: 3.9 g (15.2 mmol, 93%)
melting point: 312° C.
The following compounds are prepared analogously to Method 13:


		MS
		(ESI)
		(M + H)⁺

		169

		211

		213

		238

Method 14

2-(4-carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

a) 2-(4-benzyloxycarbonyl-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

2 g (9.217 mmol) 2,4-dichloro-5-trifluoromethylpyrimidine are dissolved in 4 ml dioxane and combined with 6.01 g (18.430 mmol) caesium carbonate and 2.16 g (7.363 mmol) benzyl 4-amino-3-methoxybenzoate (WO 9825901). This suspension is stirred for 30 h at 100° C. The suspension is combined with 50 ml dichloromethane and methanol and filtered to remove the insoluble constituents. The solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier used is silica gel and the eluant used is a mixture of 85% cyclohexane and 15% ethyl acetate.
Yield: 1.03 g (2.360 mmol; 26%)
UV max: 320 nm
MS (ESI): 438/440 (M+H)⁺ Cl distribution
436/438 (M−H)⁻ Cl distribution

b) 2-(4-carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

1 g (2.284 mmol) 2-(4-benzyloxycarbonyl-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine are dissolved in 50 ml THF and combined with 100 mg palladium hydroxide. The reaction mixture is stirred for 16 h at ambient temperature and 4 bar hydrogen pressure. Then the catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 0.76 g (2.192 mmol; 96%)
UV max: 288 nm
MS (ESI): 346/348 (M−H)⁻Cl distribution
The following compounds are prepared analogously to this process:

2-(4-carboxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

MS (ESI): 316/318 (M−H)⁻Cl distribution

2-[4-(4-benzyloxycarbonyl-piperazin-1-yl)-phenylamino]-4-chloro-5-trifluoromethyl-pyrimidine

MS (ESI): 492/494 (M+H)⁺Cl distribution

2-[4-(4-benzyloxycarbonyl-piperazin-1-yl)-2-methoxy-phenylamino]-4-chloro-5-trifluoromethyl-pyrimidine

MS (ESI): 522/524 (M+H)⁺Cl distribution

Method 15

3-pyrrolidin-1-yl-cyclobutylamine

a) tert. butyl (3-benzyloxy-cyclobutyl)-carbamate

9.28 g (45 mmol) 3-benzyloxy-cyclobutancarboxylic acid (Org. Lett. 6(11), 1853-1856, 2004) are suspended in 80 ml dry tert-butanol and combined with 5.1 g (50 mmol) triethylamine and 13.8 g (50 mmol) phosphoric acid diphenylester azide. The reaction mixture is stirred for 20 h under reflux conditions. The solvent is eliminated in vacuo and the residue is taken up in dichloromethane. The organic phase is washed three times with 2 N sodium hydroxide solution, dried with sodium sulphate and the dichloromethane is eliminated in vacuo. The crude product is recrystallised from acetonitrile (1 g crude product: 5 ml acetonitrile).
Yield: 5.98 g (22 mmol; 48%)
MS (ESI): 178 (M+H-boc)⁺ Boc cleaving in the mass detector

b) tert. butyl (3-hydroxy-cyclobutyl)-carbamate

2.77 g (10 mmol) tert. butyl (3-benzyloxy-cyclobutyl)-carbamate are suspended in 100 ml of methanol and combined with 200 mg palladium hydroxide. The reaction mixture is stirred for 5 h at 45° C. and 45 bar H₂pressure. Then the catalyst is filtered off and the solvent is eliminated in vacuo. The residue is taken up in chloroform and washed three times with aqueous sodium hydrogen carbonate solution. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo.
Yield: 1.53 g (8.2 mmol; 82%)
MS (ESI): 188 (M+H)⁺

c) tert. butyl (3-tosyl-cyclobutyl)-carbamate

18.7 g (100 mmol) tert. butyl (3-hydroxy-cyclobutyl)-carbamate and 12.1 g (120 mmol) triethylamine are placed in 500 ml chloroform. 20.5 g (105 mmol) tosyl chloride, dissolved in 150 ml chloroform, is added dropwise to this solution at 0° C. with stirring. Then the mixture is left to come up to ambient temperature and stirred for 2 h. The organic phase is washed successively with water, dilute hydrochloric acid, sodium hydrogen carbonate solution and water. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo.
Yield: 28.30 g (83 mmol; 83%)
MS (ESI): 342 (M+H)⁺

d) tert. butyl (3-pyrrolidine-cyclobutyl)-carbamate

34.1 g (100 mmol) tert. butyl (3-tosyl-cyclobutyl)-carbamate are dissolved in 750 ml pyrrolidine, and combined with a catalytic amount of DMAP. The reaction mixture is refluxed for 20 h with stirring. The pyrrolidine is eliminated in vacuo, the residue is taken up in 500 ml of ethyl acetate and washed twice with saturated sodium hydrogen carbonate solution. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo. The crude product consists—as in all the analogous reactions—of a mixture of 2 isomeric compounds which are separated by column chromatography. The stationary phase used is silica gel and the eluant used is dichloromethane, to which 9% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
The substances that elute first are designated as follows:
Yield product A: 1 g (4.17 mmol; 4%)
RF value (silica gel; dichloromethane:methanol:conc. aqueous ammonia=90:9:1)=0.62
The substances that elute second are designated as follows:
Yield product C, 2.00 g (8.33 mmol; 8%)
RF value (silica gel; dichloromethane:methanol:conc. aqueous ammonia=90:9:1)=0.53

e) (*1′,*1″)-3-pyrrolidin-1-yl-cyclobutylamine

1 g (4.17 mmol) tert. butyl (3-pyrrolidine-cyclobutyl)-carbamate (product A from precursor) are stiffed in 20 ml of a 2 N aqueous hydrochloric acid solution for 2 h at 40° C. Then the solvent is eliminated in vacuo and the residue is recrystallised from ethanol.
Yield: 0.43 g (2.786 mmol; 67%)
MS (ESI): 141 (M+H)⁺
The following compounds are prepared analogously to this process:


	MS
	(ESI)
	(M + H)⁺

	170

	210

	184

	224

	171

	212

	143

	198

	196

	194

	183

(*2′,*2″)-3-pyrrolidin-1-yl-cyclobutylamine

1 g (4.17 mmol) tert. butyl (3-pyrrolidine-cyclobutyl)-carbamate (product C from precursor) are stiffed in 20 ml of a 2 N aqueous hydrochloric acid solution for 2 h at 40° C. Then the solvent is eliminated in vacuo and the residue is recrystallised from ethanol.
Yield: 0.43 g (3.09 mmol; 74%)
MS (ESI): 141 (M+H)⁺
The following compounds are prepared analogously to this method:


	MS
	(ESI)
	(M + H)⁺

	155

	157

	171

	184

	170

	210

	253

	224

	183

	212

	143

	141

	198

	251

	194

	196

	171

Method 16

2-(4-carboxy-2-bromo-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

1 g (3.15 mmol) 2-(4-carboxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine are dissolved in 5 ml DMF and combined batchwise with 3.36 g (18.89 mmol) N-bromosuccinimide. The reaction mixture is stirred for 16 h at ambient temperature. The solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 2% water and 98% acetonitrile at the finishing point. 0.1% formic acid is added in each case to both the water and to the acetonitrile.
Yield: 0.57 g (1.44 mmol; 46%)
MS (ESI): 396/398 (M−H)⁺Cl/Br distribution

Method 17

5-amino-3-(2-fluoro-ethyl)-3H-quinazolin-4-one

500 mg (3.102 mmol) 5-amino-3H-quinazolin-4-one are combined with 2 ml (15.596 mmol) 1-bromo-2-fluoroethane. 125 mg (3.125 mmol) sodium hydride are added thereto and the mixture is stirred for 5 days at ambient temperature. The reaction mixture is diluted with 100 ml of ethyl acetate and washed with 100 ml saturated aqueous sodium chloride solution. The aqueous phase is combined with 50 ml 1 N sodium hydroxide solution and extracted 5 times with ethyl acetate. The combined organic phases are dried and the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid is added in each case to both the water and to the acetonitrile.
Yield: 67 mg (0.323 mmol; 10%)
MS (ESI): 208 (M+H)⁺

Method 18

8-amino-2-(2-fluoro-ethyl)-3,4-dihydro-2H-isoquinolin-1-one

a) 8-dibenzylamino-3,4-dihydro-2H-isoquinolin-1-one

1.466 g (9.039 mmol) 8-amino-3,4-dihydro-2H-isoquinolin-1-one are dissolved in 15 ml DMF and combined with 3.226 g (23.340 mmol) potassium carbonate and with 3.808 ml (31.420 mmol) benzylbromide. This reaction mixture is stirred for 16 h at 50° C. The reaction mixture is diluted with ethyl acetate and extracted with sodium hydrogen carbonate solution. The organic phases are dried and the solvent is eliminated in vacuo.
Yield: 1.670 g (4.877 mmol; 54%)
MS (ESI): 343 (M+H)⁺

b) 8-dibenzylamino-2-(2-fluoro-ethyl)-3,4-dihydro-2H-isoquinolin-1-one

1.06 g (3.095 mmol) 8-dibenzylamino-3,4-dihydro-2H-isoquinolin-1-one are combined with 1.5 ml (12 mmol) 1-bromo-2-fluoro-ethane and at ambient temperature 780 mg (19.50 mmol) sodium hydride are added batchwise over a period of 30 h. The reaction mixture is diluted with ethyl acetate and extracted with sodium hydrogen carbonate solution. The organic phases are dried and the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 5% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
Yield: 0.83 g (2.136 mmol; 69%)
MS (ESI): 389 (M+H)⁺

c) 8-amino-2-(2-fluoro-ethyl)-3,4-dihydro-2H-isoquinolin-1-one

830 mg (2.136 mmol) 8-dibenzylamino-2-(2-fluoro-ethyl)-3,4-dihydro-2H-isoquinolin-1-one are dissolved in 50 ml of methanol and combined with 80 mg palladium hydroxide. The reaction mixture is stirred for 48 h at ambient temperature and 4.5 bar H₂pressure. Then the catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 0.403 g (1.935 mmol; 91%)
MS (ESI): 209 (M+H)⁺
The following compounds are prepared analogously to this process:


			MS (ESI)
			(M + H)⁺

			177

			191

			223

Method 19

7-amino-5H-phenanthridin-6-one

250 mg (1.16 mmol) methyl 2-chloro-6-nitro-benzoate, 458 mg (1.392 mmol) caesium carbonate, 211 mg (1.218 mmol) 2-nitrophenylboric acid and 18 mg (0.035 mmol) bis(tri-tert-butylphosphin)palladium(0) are placed under argon and combined with 0.8 ml dioxane. This reaction mixture is stirred for 48 h at 80° C. The reaction mixture is diluted with ethyl acetate and extracted with 1 N hydrochloric acid. The organic phase is dried and the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid is added to both the water and the acetonitrile. The suitable fractions are freeze-dried. 71 mg of the intermediate product thus obtained are dissolved in 50 ml of methanol and combined with 10 mg palladium on charcoal. The reaction mixture is stirred for 48 h at ambient temperature and 4.5 bar H₂pressure. 50 ml dichloromethane are added to the reaction solution, the mixture is treated for 5 min in the ultrasound bath and then the catalyst is filtered off. The solvent is eliminated in vacuo.
Yield: 46 mg (0.221 mmol; 94%)
MS (ESI): 211 (M+H)⁺

Method 20

C-(5-morpholin-4-ylmethyl-1H-[1,2,3]triazol-4-yl)-methylamine

18.021 g (100 mmol) 1-azido-4-morpholino-2-butyne and 19.728 g (100 mmol) dibenzylamine are dissolved in 100 ml dioxane and heated to 80° C. with stiffing. After stirring for 20 h at this temperature the solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 5% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added. The suitable fractions are combined and the solvent is eliminated in vacuo. The residue is dissolved in 480 ml of methanol and combined with 30 ml concentrated aqueous hydrochloric acid and 1 g palladium on charcoal. This reaction mixture is stiffed for 5 h at 50° C. and 50 bar H₂pressure. Then the catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 8.588 g (28.00 mmol; 28%)
MS (ESI): 198 (M+H)⁺

Method 21

4-morpholin-4-ylmethyl-cyclohexylamine

2.5 g (11 mmol) tert. butyl trans-(4-formyl-cyclohexyl)-carbamate dissolved in 25 ml dimethylacetamide are combined with 1 ml (11 mmol) morpholine and 0.7 ml acetic acid. 2.4 g (11.3 mmol) sodium triacetoxyborohydride dissolved in 12.5 ml dimethylacetamide is added to this mixture. The reaction mixture is stirred for 16 h at ambient temperature. Then the reaction mixture is added to 250 ml 10% potassium hydrogen carbonate solution and this mixture is extracted three times with 100 ml of ethyl acetate. The organic phases are combined, dried and then the solvent is eliminated in vacuo. The residue is taken up in 20 ml dichloromethane and 20 ml trifluoroacetic acid and stirred for 1 h at ambient temperature. The solvents are eliminated in vacuo.
Yield: 4.22 g (9.9 mmol; 90%) (double trifluoroacetic acid salt)
MS (ESI): 199 (M+H)⁺
The following compounds are prepared analogously to this process:


	MS (ESI)
	(M + H)⁺

	157		183

	157		169

Method 22

7-amino-2-(2-fluoro-ethyl)-3-methyl-2,3-dihydro-isoindol-1-one

10 g (42.157 mmol) methyl 2-acetyl-6-nitro-benzoate (J. Org. Chem. (1952), 17, 164-76), 6.06 g (54.804 mmol) 2-fluoroethylamine and 9.32 ml (54.804 mmol) N-ethyldiisopropylamine are suspended in 25 ml of toluene and refluxed for 40 h with stirring. The reaction mixture is diluted with 400 ml of methanol and combined with 2.5 g palladium on charcoal. Then the mixture is stirred for 48 h at ambient temperature and 5 bar H₂pressure. The catalyst is filtered off and the solvent is eliminated in vacuo. The residue is taken up in dichloromethane and washed with water. The organic phase is dried with magnesium sulphate, the solvent is eliminated in vacuo and the crude product is purified by chromatography. The carrier used is silica gel and the eluant used is a mixture of cyclohexane:ethyl acetate (70:30).
Yield: 3.83 g (18.404 mmol, 43%)
MS (ESI): 209 (M+H)⁺
UV max: 318 nm
The following compounds are prepared analogously to this process, using the corresponding methyl 6-nitro-benzoate derivative:


		MS (ESI)
		(M + H)⁺

		163

		177

		203

		207

		217

		221

		227

		241

		223

		225

		239

		253

		252

		278

		237

		245

Method 23

2-azetidin-1-yl-ethylamine

500 μl (7.49 mmol) azetidin are dissolved in 15 ml acetonitrile, combined with 4.831 g (34.822 mmol) potassium carbonate and 445 μl (7.038 mmol) chloroacetonitrile. This reaction mixture is stirred for 20 h at ambient temperature. To this reaction mixture are added 20 ml diethyl ether, the suspension is stirred for 10 min and filtered to separate the solid constituents. The filtrate is freed from solvents in vacuo. 463 mg (4.816 mmol) of this intermediate product are dissolved in 50 ml 7 N methanolic ammonia and Raney nickel is added. The reaction mixture is stirred for 2 h at 60° C. and 20 bar H₂pressure. The catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 365 mg (3.664 mmol, 48%)
MS (ESI): 101 (M+H)⁺
The following compounds are prepared analogously to this process:


		MS (ESI)
		(M + H)⁺

		129

		131

		158

		159

		159

		141

		165

		172

		156

		157

		143

		145

		145

		158

		198

		145

Method 24

((S)-3-amino-pyrrolidin-1-yl)-acetonitrile

1 g (5.369 mmol) (S)-3-(Boc-amino)-pyrrolidine are dissolved in 20 ml acetonitrile and combined with 4.831 g (34.822 mmol) potassium carbonate and 322 μl (5.101 mmol) chloroacetonitrile. This reaction mixture is stirred for 20 h at ambient temperature. 20 ml diethyl ether are added to this reaction mixture, the suspension is stirred for 10 min and filtered to separate off the solid constituents. The filtrate is freed from the solvents in vacuo. The intermediate product is dissolved in 2 ml dioxane and combined with 13 ml of 4 N dioxanic hydrochloric acid and stirred overnight at RT. Then the solvent is eliminated in vacuo.
Yield: 500 mg (3.995 mmol, 74%)
MS (ESI): 126 (M+H)⁺

Method 25

(R)-2-pyrrolidin-1-yl-propylamine

a) (R)-2-pyrrolidin-1-yl-propionamide

2 g (16.055 mmol) R-alaninamide hydrochloride, 6.67 g (16.083 mmol) potassium carbonate and 8 mg (0.048 mmol) potassium iodide are suspended in 50 ml acetonitrile and then combined with 1.921 ml (16.083 mmol) 1,4-dibromobutane. This reaction mixture is refluxed for 14 h with stirring. 100 ml 1 N hydrochloric acid and 100 ml dichloromethane are added to the reaction mixture. The organic phase is separated off and discarded. The aqueous phase is made basic with sodium hydroxide solution and extracted three times with dichloromethane. The organic phases are combined, dried and freed from the solvent in vacuo.
Yield: 1.305 g (9.177 mmol, 57%)
MS (ESI): 143 (M+H)⁺

b) (R)-2-pyrrolidin-1-yl-propylamine

Under a nitrogen atmosphere 31.65 ml 1 M Lithiumaluminiumhydrid solution (THF) are taken and combined with 1 g (7.032 mmol) (R)-2-pyrrolidin-1-yl-propionamide, dissolved in 2 ml THF, at 0° C. The reaction mixture is stirred for 48 h at 50° C. The reaction mixture is combined with 100 ml of methanol and then with the same amount of dichloromethane while cooling with ice. Approx. 25 g silica gel are added to this mixture and the solvent is eliminated in vacuo. This silica gel applied to a suction filter which has previously been charged with approx. 75 g silica gel. The suction filter is washed batchwise with a total of 500 ml of a mixture of dichloromethane, methanol and aqueous conc. ammonia (90:9:1). The majority of the solvent is eliminated at a vacuum of 200 mbar and a sump temperature of approx. 50° C. The product is distilled at 69-71° C. and 10 mbar.
Yield: 160 mg (1.248 mmol, 18%)
MS (ESI): 129 (M+H)⁺
The following compounds are prepared analogously to this process:


			MS
			(ESI)
			(M + H)⁺

			129

			129

			129

			143

			157

			157

			169

			183

			183

			197

Method 26

2-chloro-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

1.1 g (5.07 mmol) 2,4-dichloro-5-trifluoromethylpyrimidin are dissolved in 1 ml dioxane and combined with 0.9 g (4.322 mmol) 7-amino-2-(2-fluoro-ethyl)-3-methyl-2,3-dihydro-isoindol-1-one (method 22) and 0.9 ml (5.257 mmol) diisopropyethylamine. This mixture is stirred for 1 h at 80° C. Then the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 20% water and 80% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are combined with dichloromethane, the organic phase is separated off, dried and the solvent is eliminated in vacuo.
Yield: 485 mg (1.250 mmol, 25%)
MS (ESI): 389/391 (M+H)⁺; Cl distribution
The following compounds are prepared analogously to this process. The aniline derivatives used are described in the supplements to method 2, in method 10 and in the supplements to method 10. The preparation of the 2,4-dichloropyrimidine derivatives is known from the literature or may be carried out by methods known from the literature.


		MS
		(ESI)
		(M + H)⁺

		363/365

		367/369

		349/351

		381/383

		333/335

		373/375

		447/449

		355/357

		399/401

		366/368

		345/347

		385/387

		381/383

Method 27

2-[2-(4-amino-3-methoxy-phenyl)-1H-imidazol-4-yl]-ethanol

a) 3-methoxy-4-nitro-benzonitrile

25 g (150.504 mmol) 3-fluoro-4-nitrobenzonitrile and 25 g (462.757 mmol) sodium methoxide are dissolved in 125 ml THF at 0° C. This reaction mixture is stirred for 30 min. The reaction mixture is extracted with ethyl acetate and 1 N hydrochloric acid. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo.
Yield: 25.092 g (140.852 mmol, 94%)
UV max: 334 nm

b) 3-methoxy-4-nitro-benzamidine

99 ml (99 mmol) lithium-bis-trimethylsilylamide solution (1 mol/l in THF) are diluted with 640 ml THF, cooled to 10° C. and combined with 8.3 g (46.591 mmol) 3-methoxy-4-nitro-benzonitrile. The reaction mixture is stirred for 10 min at 20° C. The mixture is cooled to 0° C. and combined with 80 ml 3 N hydrochloric acid. The reaction mixture is evaporated down in vacuo and extracted with water and ethyl acetate. The aqueous phase is adjusted to pH 14 with 3 N sodium hydroxide solution.
The product is then suction filtered.
Yield: 14.30 g (crude product: 60% purity)
MS (ESI): 196 (M+H)⁺
UV max: 334 nm

c) [2-(3-methoxy-4-nitro-phenyl)-1H-imidazol-4-yl]acetic acid

7 g (60% purity, 21.519 mmol) 3-methoxy-4-nitro-benzamidine are dissolved in methanol and combined with 11 ml (44 mmol) 4 N dioxanic hydrochloric acid, the solvents are eliminated in vacuo. The residue and 6.13 g (44.384 mmol) potassium carbonate are suspended in 350 ml acetonitrile and combined with 3.24 ml (22.764 mmol) ethyl 4-chloracetoacetate and 880 mg (5.301 mmol) potassium iodide. The reaction mixture is stirred for 16 h at 45° C. The reaction mixture is diluted with water and combined with 1 N sodium hydroxide solution, and extracted with ethyl acetate. The aqueous phase is adjusted to pH 1 with 1 N HCL and saturated with sodium chloride. The product is then suction filtered.
Yield: 1.45 g (5.230 mmol, 24%)
MS (ESI): 278 (M+H)⁺
UV max: 294 nm

d) 2-[2-(3-methoxy-4-nitro-phenyl)-1H-imidazol-4-yl]-ethanol

1.45 g (5.23 mmol) [2-(3-methoxy-4-nitro-phenyl)-1H-imidazol-4-yl]acetic acid are dissolved in 36 ml THF and cooled to 0° C. and combined with 10 ml (18 mmol) borane-THF complex (1.8 mol/l). After 1 h the mixture is heated to 20° C. and stirred for 16 h. Water is added until the development of gas has ended. Then the mixture is extracted twice with saturated aqueous sodium hydrogen carbonate solution and ethyl acetate. The organic phases are combined, dried and freed from the solvent in vacuo.
Yield: 0.65 g (2.465 mmol, 47%)
MS (ESI): 264 (M+H)⁺
UV max: 298 nm

e) 2-[2-(4-amino-3-methoxy-phenyl)-1H-imidazol-4-yl]-ethanol

0.144 g (0.547 mmol) 2-[2-(3-methoxy-4-nitro-phenyl)-1H-imidazol-4-yl]-ethanol are dissolved in 100 ml of methanol and combined with 0.08 g (5%) palladium on charcoal. The reaction mixture is hydrogenated for 16 h at 20° C. and 4 bar H₂pressure. The palladium on charcoal is suction filtered and the methanol is eliminated in vacuo.
Yield: 87 mg (0.373 mmol, 68%)
MS (APCI): 234 (M+H)⁺
UV max: 314 nm
The following compounds are prepared analogously to this process:


		MS (ESI)
		(M + H)⁺

		220

		251

		190

2-[2-(4-amino-3-methoxy-phenyl)-thiazole-5-yl]-ethanol is prepared analogously to the processes described above. For the cyclisation, 4-amino-3-methoxy-thiobenzamide is used (analogously to J. Am. Soc. 82, 2656, 1960) instead of 3-methoxy-4-nitro-benzamidine.
MS (ESI): 251 (M+H)⁺

Method 28

2-methoxy-N⁴-(3-pyrrolidin-1-yl-propyl)-benzene-1,4-diamine

a) (3-methoxy-4-nitro-phenyl)-(3-pyrrolidin-1-yl-propyl)-amine

1 g (5.884 mmol) 4-fluoro-2-methoxy-1-nitro-benzene, 975 mg (7.369 mmol) 1-(3-aminopropyl)pyrrolidine and 1.5 ml (8.765 mmol) diisopropyethylamine are dissolved in 5 ml dioxane and stirred for 24 h at 95° C. The solvents are eliminated in vacuo and the crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 15% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution has been added.
Yield: 1.07 g (3.827 mmol; 65%)
MS (ESI): 280 (M+H)⁺

b) 2-methoxy-N⁴-(3-pyrrolidin-1-yl-propyl)-benzene-1,4-diamine

200 mg (0.716 mmol) (3-methoxy-4-nitro-phenyl)-(3-pyrrolidin-1-yl-propyl)-amine are dissolved in 10 ml of methanol and combined with 537 μl (2.148 mmol) dioxanic hydrochloric acid and 20 mg palladium on charcoal. The reaction mixture is stirred for 1 h at ambient temperature and 5 bar H₂pressure. The catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 213 mg (0.661 mmol, 92%)
MS (ESI): 250 (M+H)⁺
The following compounds are prepared analogously to this process:


	MS
	(ESI)
	(M +
	H)⁺

	236

	307

	333

	333

	347

	333

	240

	240

	222

	208

	262

	222

	236

	168

	250

	250

	307

	307

Method 29

2-(4-carboxy-2-bromo-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine

1 g (3.148 mmol) 2-(4-carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine (method 12 or 14) are dissolved in 5 ml DMF and combined batchwise with 3.36 g (18.889 mmol) N-bromosuccinimide. This reaction mixture is stirred for 16 h at ambient temperature. Then the solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 2% water and 98% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile.
Yield: 571 mg (1.440 mmol, 46%)
MS (ESI): 396/398 (M+H)⁺

Method 30

2-(4-Acryloylamino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

a) 4-amino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

1 g (1.925 mmol) 2-(4-carboxy-2-methoxy-phenylamino)-4-[2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino]-5-trifluoromethyl-pyrimidine (prepared analogously to Example 53) are dissolved in 2 ml of toluene and combined successively with 0.43 ml (2.503 mmol) diisopropylethylamine, with 1.8 ml tert-butanol and with 0.49 ml (2.310 mmol) diphenylphosphorylazide and heated to 80° C. for 18 h. The reaction mixture is cooled, diluted with 100 ml of ethyl acetate and washed twice with 0.5 N sodium hydroxide solution. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo. The residue is taken up in dichloromethane and combined with 4 M dioxanic hydrochloric acid. The mixture is stirred for 72 h at ambient temperature. It is diluted with ethyl acetate and extracted 4 times with 1 N hydrochloric acid. The aqueous phases are combined and extracted once with ethyl acetate. The aqueous phase is made basic with sodium hydroxide solution and extracted three times with ethyl acetate. The organic phases are combined, dried and the solvent is eliminated in vacuo.
Yield: 606 mg (1.236 mmol, 64%)
MS (ESI): 491 (M+H)⁺

b) 2-(4-Acryloylamino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

311 mg (0.634 mmol) 2-(4-amino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine are dissolved in 10 ml THF and combined with 115 μl (0.824 mmol) triethylamine and 62 μl (0.761 mmol) acrylic chloride. This mixture is stirred for 1 h at ambient temperature. Then it is diluted with ethyl acetate and extracted three times with water. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo.
Yield: 340 mg (0.625 mmol, 98%)
MS (ESI): 545 (M+H)⁺
The following compounds are Prepared analogously to this Process:


	MS
	(ESI)
	(M + H)⁺

	581

	582

	659

	611

Method 31

Separation of the racemic 7-amino-2-(2-fluoro-ethyl)-3-methyl-2,3-dihydro-isoindol-1-one (method 22) into the two enantiomers

The separation is carried out by preparative chromatography under the following conditions:
column: 280×110 mm CHIRALPAK® AD 20 μm
Eluant: 95% acetonitrile/5% isopropanol (v/v)
Flow rate: 570 ml/min
Temperature: ambient temperature
The enantiomer that elutes first is known as enantiomer 1 and in the chemical formula bears the symbol *1:

Enantiomer 1

The enantiomer that elutes second is known as enantiomer 2 and in the chemical formula bears the symbol *2:

Enantiomer 2

Method 32

7-amino-3-ethyl-indan-1-one

262 mg (1.364 mmol) copper iodide are taken and heated in an argon current. Then the copper iodide is suspended in ether and cooled to −78° C. At this temperature 0.8 ml of a 3 Methylmagnesium bromide solution (in ether) are added and the mixture is stirred for 10 min and then left to thaw to 0° C. After 15 min stirring at this temperature the mixture is cooled to −78° C. again and 200 mg (0.802 mmol) N-(3-oxo-3H-inden-4-yl)-benzamide, dissolved in 9 ml THF, are added dropwise and the mixture is stirred for 1 h at 0° C. The reaction mixture is diluted with dichloromethane and washed three times with concentrated aqueous ammonia solution. The organic phase is dried with magnesium sulphate and the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 98% water and 2% acetonitrile at the starting point and 2% water and 98% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are freeze-dried. This intermediate product is dissolved in 2 ml dioxane and combined with 5 ml concentrated hydrochloric acid. The reaction mixture is refluxed for 24 h with stirring. Then it is diluted with water and extracted three times with dichloromethane. The combined organic phases are again washed with water, dried and the solvent is removed. The residue is purified by column chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 5% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
Yield: 70 mg (0.399 mmol; 29%)
MS (ESI): 176 (M+H)⁺
The following compounds are prepared analogously to this process:


		MS (ESI)
		(M + H)⁺

		162

		190

Method 33

7-amino-3,3-dimethyl-3H-isobenzofuran-1-one

250 mg (0.609 mmol) methyl 2-dibenzylamino-benzoate are combined under argon with 0.609 ml of a 1 M lithium chloride solution (THF). This solution is cooled to −ambient temperature and slowly 0.914 ml (1.827 mmol) of a 2 M isopropyl-magnesium chloride solution are metered in. After stirring for 16 h at this temperature, 45 μl (0.609 mmol) acetone are added dropwise and the mixture is stirred for 4 h at ambient temperature. The reaction solution is combined with sodium hydrogen carbonate solution and extracted three times with dichloromethane. The combined organic phases are dried and the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are freeze-dried. This intermediate product is dissolved in 50 ml of methanol combined with 10 mg palladium on charcoal and hydrogenated for 20 h at 5 bar hydrogen pressure and ambient temperature. Then the catalyst is filtered off and the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are freeze-dried.
Yield: 34 mg (0.192 mmol; 32%)
MS (ESI): 178 (M+H)⁺
The following compounds are prepared analogously to this process:


		MS (ESI)
		(M + H)⁺

		164

		192

		220

		190

		178

Method 34

7-amino-2-(2-fluoro-ethyl)-3,3-dimethyl-2,3-dihydro-isoindol-1-one

a) methyl 2-(cyano-dimethyl-methyl)-6-nitro-benzoate

3 g (13.625 mmol) methyl 2-cyanomethyl-6-nitro-benzoate (WO 9518097) are dissolved in 20 ml THF combined with 4.33 ml (68.788 mmol) iodomethane and cooled to 0° C. At this temperature 40.87 ml of a 1 M potassium-tert-butoxide solution is slowly added dropwise. The mixture is heated to ambient temperature and stirred for 16 h at this temperature. The reaction mixture is diluted with ethyl acetate and extracted three times with 1 M hydrochloric acid. The combined organic phases are dried and the solvent is eliminated in vacuo.
Yield: 3.11 g (12.535 mmol; 92%)

b) 3,3-dimethyl-7-nitro-2,3-dihydro-isoindol-1-one

Reaction mixture 1
1 g (4.028 mmol) methyl 2-(cyano-dimethyl-methyl)-6-nitro-benzoate are suspended in 20% ethanolic potassium hydroxide solution and stiffed for 24 h at ambient temperature.

Reaction Mixture 2

1.9 g (47.577 mmol) sodium hydroxide are dissolved in 40 ml of water cooled to 0° C. and combined with 0.5 ml (28.899 mmol) bromine reaction mixture 1 is slowly added dropwise to this solution. After 8 h the same amount of reaction mixture 1 is added again. The mixture is stiffed for a further 48 h at RT. Then sodium sulphite solution is added, the mixture is stirred for 20 min and then acidified with potassium hydrogen sulphate solution. It is extracted three times with ethyl acetate. The combined organic phases are dried and the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is silica gel and the eluant used is a mixture of cyclohexane:ethyl acetate (3:1).
Yield: 67 mg (0.325 mmol, 8%)
MS (ESI): 207 (M+H)⁺

c) 3,3-dimethyl-7-amino-2,3-dihydro-isoindol-1-one

67 mg (0.325 mmol) 3,3-dimethyl-7-nitro-2,3-dihydro-isoindol-1-one are dissolved in 50 ml of methanol and combined with 10 mg palladium on charcoal. The mixture is hydrogenated for 16 h at 4 bar H₂pressure and ambient temperature. Then the catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 50 mg (0.284 mmol, 93%)
MS (ESI): 177 (M+H)⁺

d) 7-dibenzylamino-3,3-dimethyl-2,3-dihydro-isoindol-1-one

50 mg (0.284 mmol) 3,3-dimethyl-7-amino-2,3-dihydro-isoindol-1-one are dissolved in 0.5 ml DMF and combined with 141 mg (1.021 mmol) potassium carbonate and 10 mg (0.028 mmol) tetrabutylammonium iodide. The mixture is heated to 50° C. and 155 μl (1.277 mmol) benzylbromide are added dropwise thereto. After stirring for 16 h at this temperature the mixture is diluted with ethyl acetate and extracted three times with 1 M hydrochloric acid. The combined organic phases are dried and the solvent is eliminated in vacuo.
Yield: 67 mg (0.188 mmol; 66%)
MS (ESI): 357 (M+H)⁺
e) 7-dibenzylamino-2-(2-fluoro-ethyl)-3,3-dimethyl-2,3-dihydro-isoindol-1-one 67 mg (0.188 mmol) 7-dibenzylamino-3,3-dimethyl-2,3-dihydro-isoindol-1-one are dissolved in 1 ml (7.877 mmol) 1-bromo-2-fluoroethane and combined with 52 mg (0.376 mmol) sodium hydride. After 4 h stirring at ambient temperature the mixture is diluted with ethyl acetate and extracted three times with 1 M hydrochloric acid. The combined organic phases are dried and the solvent is eliminated in vacuo.
Yield: 75 mg (0.188 mmol; 100%)
MS (ESI): 403 (M+H)⁺

f) 7-amino-2-(2-fluoro-ethyl)-3,3-dimethyl-2,3-dihydro-isoindol-1-one

75 mg (0.188 mmol) 7-dibenzylamino-2-(2-fluoro-ethyl)-3,3-dimethyl-2,3-dihydro-isoindol-1-one are dissolved in 50 ml of methanol and combined with 10 mg palladium on charcoal. The mixture is hydrogenated for 16 h at 5 bar H₂pressure and ambient temperature. Then the catalyst is filtered off and the solvent is eliminated in vacuo.
Yield: 36 mg (0.162 mmol, 87%)
MS (ESI): 223 (M+H)⁺

Example 1

2-(2-methoxy-4-N-propylcarbamoyl-phenylamino)-4-(3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

100 mg (0.257 mmol) 2-(2-methoxy-4-propylcarbamoyl-phenylamino)-4-chloro-5-trifluoro-methyl-pyrimidine (method 1) are dissolved in 1 ml N,N-dimethylacetamide and combined with 83 mg (0.565 mmol) 7-amino-2,3-dihydro-isoindol-1-one (method 2). 48 μl of a 4 molar solution of HCl (0.193 mmol) in 1,4-dioxane are metered into this reaction mixture. After two days at 50° C. the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 5% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added. The concentrated crude product is again purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 80% water and 20% acetonitrile at the starting point and 60% water and 40% acetonitrile at the finishing point.
Yield: 42 mg (0.084 mmol; 33%)
UV max: 318 nm
MS (ESI): 501 (M+H)⁺
¹H-NMR: 0.92 (t, 3H), 1.51-1.63 (m, 2H), 3.21-3.29 (m, 2H), 3.86 (s, 3H), 4.37 (s, 2H), 7.14-7.21 (m, 1H), 7.33 (t, 1H), 7.47-7.54 (m, 1H), 7.55-7.60 (m, 1H), 7.73-7.82 (m, 1H), 8.35-8.50 (m, 3H), 8.75 (s, 1H), 9.09 (s, 1H), 10.66 (s, 1H)

Examples 2-17

The following compounds are prepared by an analogous method as described in Example 1. 2-(2-Methoxy-4-propylcarbamoyl-phenylamino)-4-chloro-5-trifluoromethylpyrimidine and a corresponding 7-amino-2,3-dihydro-isoindol-1-one derivative (method 2) are used. N-methyl-2-pyrrolidinone or N,N-dimethylacetamide is used as solvent.

		UV max	MS (ESI)
#	A	[nm]	(M + H)⁺

2	322	515

3	314	529

4	285	543

5	286/310	583

6	322	571

7	285/321	585

8	285/318	559

9	285/318	586

10	281/316	626

11	284/316	545

12	325	577

13	282/318	595

14	284/322	573

15	286, 306	607

16	325

17	318/282	607

Example 18

2-(2-methoxy-4-N-propylcarbamoyl-phenylamino)-4-(3-oxo-1,3-dihydro-isobenzofuran-4-ylamino)-5-trifluoromethyl-pyrimidine

100 mg (0.257 mmol) 2-(2-methoxy-4-propylcarbamoyl-phenylamino)-4-chloro-5-trifluoro-methyl-pyrimidine (method 1) are dissolved in 1 ml N,N-dimethylacetamide and combined with 46 mg (0.308 mmol) 7-amino-3H-isobenzofuran-1-one (Safdar Hayat et al., Tetrahedron Lett 2001, 42(9):1647-1649). 48 μl of a 4 molar solution of HCl (0.193 mmol) in 1,4-dioxane zudosiert metered into this reaction mixture. After 4 days at 50° C. the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 4% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
Yield: 26 mg (0.051 mmol; 20%)
UV max: 322 nm
MS (ESI): 502 (M+H)⁺
¹H-NMR: 0.92 (t, 3H), 1.51-1.63 (m, 2H), 3.22-3.28 (m, 2H), 3.86 (s, 3H), 5.42 (s, 2H), 7.24-7.30 (m, 1H), 7.44-7.55 (m, 2H), 7.55-7.60 (m, H), 7.67-7.78 (m, 1H), 8.38-8.48 (m, 2H), 8.50 (s, 1H), 9.21 (s, 1H), 9.64 (s, 1H)

Examples 19-37

The following compounds are prepared by analogous methods to those described in Example 1 and Example 18. 2-(2-methoxy-4-propylcarbamoyl-phenylamino)-4-chloro-5-trifluoromethylpyrimidine (method 1) is used. The corresponding aniline derivative is commercially obtainable, known from the literature or is prepared by the processes described in method 2 and 4 to 9. N-methyl-2-pyrrolidinone or N,N-dimethylacetamide is used as solvent.

		UV max	MS (ESI)
#	A	[nm]	(M + H)⁺

19		235	586

20		323/226	543

21		325	530

22		262	514

23		320	544

24		318	542

25		312	530

26		315	529

27		314	528

28		317	502

29		316	516

30		322	529

31		255	548

32		320	500

33		325	515

34		250/286/ 318	516

35		320	558

36		316	514

37		321

Example 38

2-(2-methoxy-4-N-propylcarbamoyl-phenylamino)-4-(4-methyl-5-oxo-2,3,4,5-tetrahydro-benzo[f][1,4]oxazepin-6-ylamino)-5-trifluoromethyl-pyrimidine

50 mg (0.129 mmol) 2-(2-methoxy-4-propylcarbamoyl-phenylamino)-4-chloro-5-trifluoro-methyl-pyrimidine (method 1) are dissolved in 200 μl 1,4-dioxane and combined with 25 mg (0.13 mmol) 6-amino-4-methyl-3,4-dihydro-2H-benzo[f][1,4]oxazepin-5-one (method 10). 36 μl of a 4 molar solution of HCl (0.144 mmol) in 1,4-dioxane are metered into this reaction mixture. After 4 days at 50° C. the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is a mixture of dichloromethane and ethyl acetate (1:1).
Yield: 23 mg (0.042 mmol; 33%)
UV max: 318 nm
MS (ESI): 545 (M+H)⁺
¹H-NMR: 0.91 (t, 3H), 1.49-1.61 (m, 2H), 3.09 (s, 3H), 3.20-3.28 (m, 2H), 3.49 (t, 2H), 3.88 (s, 3H), 4.31 (t, 2H), 6.83-6.88 (m, 1H), 7.34-7.45 (m, 2H), 7.50-7.54 (m, 1H), 7.88-8.00 (m, 2H), 8.37-8.44 (m, 2H), 8.62 (s, 1H), 9.97 (s, 1H)

Examples 39-52

The following compounds are prepared by analogous methods to those described in Example 1 and 18. 2-(2-methoxy-4-propylcarbamoyl-phenylamino)-4-chloro-5-trifluoromethylpyrimidine (method 1) is used. The corresponding aniline derivative is commercially obtainable, known from the literature or is prepared by the processes described in method 10 and 11. N-methyl-2-pyrrolidinone or N,N-dimethylacetamide is used as solvent.

		UV max	MS (ESI)
#	A	[nm]	(M + H)⁺

39		229/279/ 315	559

40		282/314	545

41		282/318	587

42		282/314	571

43		282/318	585

44		318	559

45		234/320	559

46		282/218	603

47		278/318	531

48		286/314	573

49		274/314	558

50		318	587

51		223/282/318	579

52		318	634

Example 53

2-[2-methoxy-4-(4-morpholin-4-yl-(1,4-trans-cyclohexyl)carbamoyl)-phenylamino]-4-(2-carbamoyl-3-fluoro-phenylamino)-5-trifluoromethyl-pyrimidine

102 mg (0.29 mmol) 2-(4-carboxyamino-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine (method 12) are dissolved in 250 μl N-methyl-2-pyrrolidinone and combined with 47 mg (0.319 mmol) 7-amino-indan-1-one (method 8). 15 μl of a 4 M solution of HCl (0.058 mmol) in 1,4-dioxane are metered into this reaction mixture. After 16 h at 90° C. the reaction mixture is stirred into 150 ml of a aqueous 1 N hydrochloric acid. The precipitate is filtered off and dried in vacuo. 100 mg (0.174 mmol) of this precipitate, 150 μl (0.875 mmol) N-ethyldiisopropyl-amine, 68 mg (0.210 mmol) O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-tetrafluoroborate and 30 mg (0.163 mmol) trans-4-morpholin-4-yl-cyclohexylamine (method 13) are dissolved in 5 ml N,N-dimethylformamide. After 15 h at ambient temperature the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier used is silica gel and the eluant used is dichloromethane, to which 7% of a mixture of 90% methanol and 10% saturated aqueous ammonia solution have been added.
Yield: 55 mg (0.100 mmol; 57%)
UV max: 318 nm
MS (ESI): 555 (M+H)⁺
¹H-NMR: 1.55-1.69 (m, 2H), 1.74-1.84 (m, 2H), 1.91-2.02 (m, 2H), 2.18 (s, 3H), 2.69-2.75 (m, 2H), 2.75-2.84 (m, 2H), 3.03-3.10 (m, 2H), 3.70-3.83 (m, 1H), 3.86 (s, 3H), 7.15-7.21 (m, 1H), 7.36-7.46 (m, 1H), 7.48-7.54 (m, 1H), 7.54-7.58 (m, 1H), 7.71-7.79 (m, 1H), 8.18-8.25 (m, 1H), 8.30-8.45 (m, 1H), 8.48 (s, 1H), 9.16 (s, 1H), 10.59 (s, 1H)

Examples 54-77

The following compounds are prepared by an analogous method to that described in Example 53. The corresponding aniline is described in method 2, 7, 8, or 9 or known from the literature. The amine used to prepare the amide is commercially obtainable or is described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

54			318	555

55			318	569

56			322	570

57			320	640

58			284, 322	556

59			282, 318	626

60			325	655

61			325	585

62			254, 286, 318	639

63			321	631

64			322	570

65			322	640

66			322	683

67			322	613

68			286, 322	654

69			286, 322	584

70			282, 322	627

71			322	670

72			286, 322	600

73			322	684

74			286, 322	614

75			322	557

76			330	732

77			325	654

Examples 78-140

The following compounds are prepared by an analogous method to that described in Example 53. 2-(4-Carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be prepared according to method 12 or 14. The corresponding aniline is described in the supplements to method 10. The amine used to prepare the amide is commercially obtainable or is described in method 13, in the supplements to method 13, 15 or 25.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

78	318	308

79	326	346

80	318	706

81	318	584

82	318	614

83	318	776

84	318	626

85	318	348

86	318	718

87	318	684

88	318	353

89	322	346

90	318	686

91	310	621

92	318	746

93	318	676

94	318	316

95	318	696

96	282; 310	571

97	318	614

98	318	684

99	315	559

100	314	621

101	314	676

102	318	747

103	318	656

104	318	586

105	318	(M

106	318	730

107	322	674

108	318	640

109	322	640

110	282, 318	614

111	226, 282, 318	640

112	318	614

113		626

114	318	640

115	318	640

116	318	654

117	318	668

118	318	628

119	318	600

120	318-322	614

121	318	670

122	318	654

123	318	626

124	282, 318	668

125	282, 318	642

126	282, 318	693

127	318	680

128	318	654

129	318	705

130	226, 282, 318	628

131	318	668

132	318-322	642

133	318	693

134	318-322	642

135	318	682

136	318	698

137	318-322	656

138	318-322	707

139	318-322	640

140	318-322	628

Examples 141-166

The following compounds are prepared by an analogous method to that described in Example 53. The preparation of 2-(4-carboxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine is described in method 14. The corresponding aniline is described in method 10. The amine used to prepare the amide is commercially obtainable or is described in method 13, 15 or 25.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

141			302	596

142			302	610

143			302	596

144			302	584

145			302	610

146			302	638

147			298	654

148			302	610

149			302	650

150			298-302	666

151			302	584

152			302	624

153			298-302	640

154			302	598

155			298-302	649

156			302	598

157			302	638

158			298-302	654

159			302	612

160			302	663

161			302	612

162			302	652

163			298-302	668

164			302	677

165			302	626

166			302	624

Example 167

2-(2-methoxy-4-piperazin-1-yl-phenylamino)-4-(3,3-dimethyl-5-oxo-2,3,4,5-tetrahydro-benzo[f][1,4]oxazepin-6-ylamino)-5-trifluoromethyl-pyrimidine

500 mg (0.958 mmol) 2-[4-(4-benzyloxycarbonyl-piperazin-1-yl)-phenylamino]-4-chloro-5-trifluoromethyl-pyrimidine (method 14) are dissolved in 0.5 ml NMP, combined with 198 mg (0.960 mmol) 6-amino-3,3-dimethyl-3,4-dihydro-2H-benzo[f][1,4]oxazepin-5-one (method 10) and with 25 μl (0.1 mmol) dioxanic hydrochloric acid. This reaction mixture is stirred for 1.5 h at 100° C. The solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile.
Yield: 0.59 g (0.86 mmol; 90%)
0.59 g (0.86 mmol) of the above-mentioned intermediate products are dissolved in 50 ml of dimethylformamide and combined with a quantity of distilled water such that there is no precipitation. To this solution are added 60 mg palladium on charcoal and the mixture is hydrogenated at 7 bar H₂pressure and 20° C. for 6 h. The catalyst is filtered off and the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists at the starting point of 60% water and 40% acetonitrile and at the finishing point of 15% water and 85% acetonitrile. 10 mmol/l ammonium hydrogen carbonate and 20 mmol/l ammonia are dissolved in the water. The suitable fractions are freeze-dried. The residue is dissolved in acetonitrile and combined with 2 ml of a 1 M hydrochloric acid solution. Then the solvent is eliminated in vacuo. The substance is obtained as the dihydrochloride.
Yield: 0.46 g (0.73 mmol; 85%)
UV max: 284 nm
MS (ESI): 558 (M+H)⁺
¹H-NMR: 1.19 (s, 6H), 3.19-3.28 (m, 4H), 3.41-3.49 (m, 4H), 3.80 (s, 3H), 4.07 (s, 1H), 6.54-6.60 (m, 1H), 6.72-6.76 (m, 1H), 6.83-6.89 (m, 1H), 7.21-7.42 (m, 2H), 7.85-8.20 (m, 1H), 8.33-8.60 (m, 1H), 8.74 (s, 1H), 9.30-9.71 (m, 3H), 12.84 (s, 1H)

Example 168

2-(2-methoxy-4-piperazin-1-yl-phenylamino)-4-(S)-4-oxo-2,3,10,10a-tetrahydro-1H,4H-9-oxa-3a-aza-benzo[f]azulen-5-ylamino-5-trifluoromethyl-pyrimidine

This compound is prepared analogously to Example 167. The aniline used is described in method 10.
Yield: 0.23 g (0.41 mmol; 91%)
UV max: 282 nm
MS (ESI): 570 (M+H)⁺
¹H-NMR: 1.53-1.71 (m, 1H), 1.79-2.06 (m, 3H), 3.15-3.32 (m, 4H), 3.32-3.55 (m, 5H), 3.58-3.72 (m, 1H), 3.72-3.94 (m, 4H), 4.00-4.23 (m, 2H), 6.48-6.61 (m, 1H), 6.68-6.77 (m, 1H), 6.83-7.00 (m, 1H), 7.19-7.50 (m, 2H), 7.78-8.10 (m, 1H), 8.23-8.60 (m, 1H), 9.18-9.64 (m, 3H), 10.54-10.86 (m, 1H)

Example 169

2-[4-(4-ethyl-piperazin-1-yl)-2-methoxy-phenylamino]-4-((S)-4-oxo-2,3,10,10a-tetrahydro-1H,4H-9-oxa-3a-aza-benzo[f]azulen-5-ylamino-5-trifluoromethyl-pyrimidine

60 mg (0.11 mmol) 2-(2-methoxy-4-piperazin-1-yl-phenylamino)-4-(S)-4-oxo-2,3,10,10a-tetrahydro-1H,4H-9-oxa-3a-aza-benzo[f]azulen-5-ylamino-5-trifluoromethyl-pyrimidine (Example 168) are dissolved in 300 μl dimethylformamide and combined with 12 μl (0.21 mmol) acetaldehyde and 47 mg (0.21 mmol) sodium triacetoxyborohydride. This reaction mixture is stirred at 20° C. for 20 h. The solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and 50% water and 50% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are combined with 500 μl of a 1 N hydrochloric acid and freeze-dried. The product is obtained as the dihydrochloride.
Yield: 49 mg (0.074 mmol; 71%)
UV max: 282 nm
MS (ESI): 598 (M+H)⁺
¹H-NMR: 1.23-1.37 (m, 3H), 1.57-1.72 (m, 1H), 1.80-2.06 (m, 3H), 3.02-3.27 (m, 6H), 3.34-3.48 (m, 1H), 3.48-3.71 (m, 3H), 3.71-3.94 (m, 7H), 6.48-6.61 (m, 1H), 6.68-6.79 (m, 1H), 6.84-6.97 (m, 1H), 7.18-7.43 (m, 2H), 7.78-8.08 (m, 1H), 8.26-8.53 (m, 1H), 9.14-9.44 (m, 1H), 10.49-10.74 (m, 1H), 10.80-11.08 (m, 1H)

Example 170

2-[4-(4-methyl-piperazin-1-yl)-2-methoxy-phenylamino]-4-((S)-4-oxo-2,3,10,10a-tetrahydro-1H,4H-9-oxa-3a-aza-benzo[f]azulen-5-ylamino-5-trifluoromethyl-pyrimidine

To prepare this compound formaldehyde is used instead of acetaldehyde. Otherwise the method is as in Example 169.
Yield: 16 mg (0.024 mmol; 28%)
UV max: 278 nm
MS (ESI): 584 (M+H)⁺
¹H-NMR: 1.58-1.71 (m, 1H), 1.81-2.06 (m, 3H), 2.78-2.88 (m, 3H), 3.00-3.23 (m, 4H), 4.03-4.21 (m, 2H), 6.48-6.59 (m, 1H), 6.69-6.78 (m, 1H), 6.80-6.91 (m, 1H), 7.17-7.44 (m, 2H), 7.92-8.15 (m, 1H), 8.34 (s, 1H), 8.86-9.04 (m, 1H), 10.38-10.64 (m, 2H)

Examples 171-180

The following Examples are prepared analogously to to Example 169 and 170. The corresponding aniline is described in the supplements to method 10.

				MS
				(ESI)
			UV max	(M +
#	A	D	[nm]	H)⁺

171			226, 282	572

172			250, 282	586

173			250, 282	596

174			250, 282	600

175			282	544

176			282	558

177			218; 282	586

178			282	582

179			226	558

180			226	572

Examples 181-332

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-Carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be obtained according to method 12 or 14. The corresponding aniline is described in method 11. The amine used to prepare the amide is commercially obtainable or described in method 13, 15 and 25.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

181			318, 282, 234	380

182			238	639

183			234; 318	709

184			318, 282, 248	558

185			318, 280	613

186			316, 282, 234	342

187			318, 284, 238	307

188			318, 282, 242	342

189			314, 282, 242	600

190			318, 282, 234	328

191			318	363

192			318, 230	650

193			314	634

194			318	634

195			318	671

196			318, 230	380

197			314, 282, 250	558

198			319	705

199			318, 226	775

200			318	634

201			314	634

202			230; 318	584

203			317	572

204			318, 230	697

205			318, 234	544

206			318	669

207			318, 230	650

208			317	627

209			318, 230	599

210			318, 230	705

211			230; 322	653

212			230; 322	655

213			230; 318	669

214			230, 282, 314	634

215			318	655

216			318, 234	725

217			314, 235	586

218			318, 230	641

219			318, 226	711

220			318, 230	640

221			318	765

222			318	600

223			315	673

224			319, 226	728

225			318, 226	798

226			318, 234	655

227			230; 322	653

228			230; 318	682

229			234; 318	639

230			318, 226	695

231			234, 282, 318	598

232			230, 282, 318	653

233			234, 282, 318	723

234			318, 222	673

235			318	725

236			318, 282, 226	798

237			230; 318	641

238			230; 318	711

239			234; 318	586

240			318, 226	745

241			322	703

242			320, 226	732

243			321, 221	694

244			230, 282, 318	652

245			234, 282, 318	707

246			230, 282, 318	777

247			230, 282, 318	630

248			234, 282, 318	685

249			234, 282, 318	755

250			230, 282, 318	630

251			230, 282, 318	685

252			230, 282, 318	755

253			230; 318	695

254			230; 318	70

255			230; 318	389

256			230; 318	652

257			230	357

258			230	784

259			230	659

260			319, 230	689

261			322	703

262			322	705

263			320	719

264			226	690

265			226; 318	760

266			230	635

267			230; 318	381

268			318	812

269			318	652

270			318	707

271			318, 226	777

272			318	659

273			318	714

274			315, 239	669

275			319, 222	723

276			318, 226	793

277			316	620

278			318	675

279			318, 226	745

280			317, 226	620

281			318	675

282			318, 230	745

283			318	784

284			318	758

285			318	688

286			238, 282, 314	616

287			230, 282, 318	671

288			230, 282, 318	741

289			234, 282, 318	616

290			226, 282, 318	671

291			234, 282, 318	741

292			234, 282, 318	648

293			230, 282, 318	703

294			226, 282, 318	773

295			226, 282, 318	893

296			226, 282, 318	727

297			226, 282, 318	754

298			230, 282, 318	823

299			282, 318	669

300			282, 318	613

301			282, 318	641

302			286, 318	639

303			286, 318	627

304			286, 318	655

305			286, 318	667

306			286, 318	717

307			286, 318	689

308			286, 318	665

309			230, 286, 318	653

310			230, 282, 318	715

311			286, 322	695

312			234, 286, 318	667

313			230, 282, 318	639

314			230, 282, 318	667

315			230, 282, 318	681

316			230, 282, 318	695

317				679

318			226, 284, 318	681

319			230, 284, 318	697

320			226, 284, 314	750

321			230, 286, 318	669

322			230, 282, 318	693

323			230, 282, 314	709

324			230, 286, 314	681

325			226, 286, 314	762

326			230, 282, 318	681

327			230, 282, 314	697

328			234, 282, 318	627

329			226, 282, 318	767

330			226, 282, 318	725

331			230, 286, 318	711

332			226, 282, 318	671

333			234, 282, 314	718

334			234, 282, 318	693

335			234, 286, 318	653

336			284, 318	706

337			230, 282, 318	641

338			230, 282, 314	667

339			283, 318	655

340			230, 286, 318	699

341			230, 282, 318	750

342			230, 282, 318	627

343			250, 282, 318	667

344			230, 282, 318	683

345			238, 282, 314	641

346			230, 314	692

347			282, 318	723

348			234, 286, 314	653

349			286, 318	667

350			234, 286, 314	718

351			230, 286, 318	685

Examples 352-372

The following compounds are prepared by an analogous process to that described in Example 53 described, prepared. 2-(4-carboxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may after method 14 prepared are. The corresponding aniline is in method 11 described. The amine used to prepare the amide is commercially obtainable or is in method 13, 15 or 25 described.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

352			222, 302	688

353			246, 298	663

354			234, 298	679

355			234, 302	623

356			298	611

357			246, 302	676

358				651

359				667

360			246, 302	611

361			298	662

362				637

363			234, 298	653

364			226, 302	597

365			302	637

366			246, 302	625

367			302	695

368			302	711

369			302	669

370			302	720

371			300	693

372			242, 302	655

Examples 373-386

The following Examples are prepared analogously to Example 169 and 170. The corresponding aniline is described in method 11.

			UV max	MS (ESI)
#	A	D	[nm]	(M + H)⁺

373			246	621

374			246	611

375			234	639

376			238	597

377			250	599

378			250	585

379			250	613

380			250	609

381			246	625

382			250	599

383			230	571

384			246	595

385			250	585

386			246, 286	615

Examples 387-388

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-Carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be prepared according to method 12 or 14. The corresponding aniline is described in method 4 or method 17. The amine used to prepare the amide is commercially obtainable.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

387			262, 318	569

388			278, 318	615

Examples 389-404

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-Carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be prepared according to method 12 or 14. The corresponding aniline is described in method 7, in method 18 or 19. The amine used to prepare the amide is commercially obtainable or is described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

389			284, 322	668

390			230, 285, 325	698

391			280, 325	730

392			230, 285, 325	682

393			285, 325	630

394			284, 322	686

395			285, 325	616

396			285, 322	654

397			285, 325	584

398			285, 325	598

399			285, 325	668

400			285, 325	598

401			285, 325	612

402			285, 322	700

403			285, 322	630

404			262	688

Example 405

2-[4-([1,4′]bipiperidinyl-4-ylcarbamoyl)-2-methoxy-phenylamino]-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

1150 mg (3.308 mmol) 2-(4-carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine (method 12 or 14) are dissolved in 2.5 ml N-methyl-2-pyrrolidinone and combined with 883 mg (4.161 mmol) 7-amino-2-(2,2-difluoro-ethyl)-2,3-dihydro-isoindol-1-one (method 2). 115 μl of a 4 M solution of HCl (0.460 mmol) in 1,4-dioxane are metered into this reaction mixture. After 16 h at 90° C. the reaction mixture is stirred into 150 ml of an aqueous 1 N hydrochloric acid. The precipitate is filtered off and dried in vacuo.
Yield: 1626 mg (3.110 mmol; 94%)
MS (ESI): 524 (M+H)⁺
100 mg (0.191 mmol) of this precipitate, 240 μl (1.402 mmol) N-ethyldiisopropyl-amine, 89 mg (0.279 mmol) O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-tetrafluoroborate and 76 mg (0.267 mmol) tert-butyl 4-amino-[1,4′]bipiperidinyl-F-carboxylate are dissolved in 3 ml N,N-dimethylformamide. After 15 h at 20° C. the solvent is eliminated in vacuo. The residue is taken up in 20 ml dichloromethane and 5 ml of methanol and filtered through aluminium oxide. The aluminium oxide is washed several times with a mixture of dichloromethane and methanol (4:1). The solvent of the combined fractions is eliminated in vacuo. The residue is dissolved in 5 ml dichloromethane and combined with 5 ml trifluoroacetic acid. This mixture is stirred for 3 h at 20° C. and then the solvent is eliminated in vacuo. The crude product is purified by column chromatography. The carrier material used is C18-RP-silica gel and a gradient is run through which consists of 90% water and 10% acetonitrile at the starting point and 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid are added both to the water and to the acetonitrile. The suitable fractions are combined with 500 μl of a 1 N hydrochloric acid and freeze-dried. The product is obtained as the trihydrochloride.
Yield: 42 mg (0.053 mmol; 28%)
UV max: 322 nm
MS (ESI): 689 (M+H)⁺
¹H-NMR: 1.92-2.19 (m, 6H), 2.28-2.37 (m, 2H), 2.86-3.00 (m, 2H), 3.07-3.19 (m, 3H), 3.84-4.18 (m, 7H), 4.59 (s, 2H), 6.15-6.47 (m, 1H), 7.23-7.28 (m, 1H), 7.35-7.43 (m, 1H), 7.54-7.64 (m, 2H), 7.75-7.82 (m, 1H), 8.40-8.64 (m, 3H), 8.90-9.01 (m, 1H), 9.10-9.25 (m, 2H), 10.40-10.47 (m, 1H), 10.91-11.27 (m, 1H)

Examples 406-407

The following compounds are prepared by an analogous process to that described in Example 405.


	UV max	MS (ESI)
#	[nm]	(M + H)⁺

406	318	606

407	322, 286	606

Example 408

2-[2-methoxy-4-(1′-methyl-[1,4′]bipiperidinyl-4-ylcarbamoyl)-phenylamino]-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

70 mg (0.087 mmol) 2-[4-([1,4′]bipiperidinyl-4-ylcarbamoyl)-2-methoxy-phenylamino]-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine (Example 405) are dissolved in 3 ml of methanol, and combined with 8.5 μl (0.508 mmol) acetic acid and with 8 μl (0.107 mmol) of a 37% aqueous formaldehyde solution. Then at 20° C. 7.0 mg (0.112 mmol) sodium cyanoborohydride are added. This mixture is stirred for 16 h at 20° C. The solvent is eliminated in vacuo and the crude product is purified by column chromatography. The carrier material used is C18-RP-silica gel and a gradient is run through which consists at the starting point of 95% water and 5% acetonitrile and at the finishing point of 5% water and 95% acetonitrile. 0.1% formic acid are added both to the water and to the acetonitrile. The suitable fractions are combined with 500 μl of a 1 N hydrochloric acid and freeze-dried. The product is obtained as the trihydrochloride.
Yield: 18 mg (0.022 mmol; 25%)
UV max: 322 nm
MS (ESI): 703 (M+H)⁺

Examples 409-491

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-Carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be prepared according to method 12 or 14. The corresponding aniline is described in method 2. The amine used to prepare the amide is commercially obtainable or is described in method 13, 20 or 21.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

409			285, 320	584

410			322	716

411			326	703

412				558

413			282, 318	699

414			322, 286	668

415			322.3	724

416			322.3	362

417			322, 286	738

418			322, 286	738

419			282, 314	738

420			286, 314	738

421			286, 318	700

422			286, 322	698

423			286, 318	700

424			286, 322	712

425			286, 322	724

426			322, 286	672

427			282, 322	723

428			322, 285	602

429			326.3	616

430			322, 286	616

431			318, 286	645

432			321, 284	632

433			322, 286	618

434			318, 282	690

435			322, 282	708

436			322, 286	686

437			322, 284	722

438			322, 282	658

439			322, 285	547

440			322, 286	602

441			286.3	565

442			322, 286	620

443			322, 284	686

444			326.3	634

445			326, 286	634

446			322, 284	676

447			322.3	663

448			325.3	650

449			325.3	635

450			322, 282	620

451			322, 282	704

452			322, 282	665

453			326, 282	595

454			322, 284	677

455			322.3	664

456			326, 286	594

457			322, 282	743

458			326, 286	638

459			326, 283	681

460			318, 284	681

461			318, 286	627

462			322, 286	627

463			326, 286	648

464			322, 286	611

465			322, 286	723

466			322, 282	710

467			326, 286	654

468			326, 286	654

469			322, 284	683

470			326, 286	640

471			318, 283	710

472			326, 286	654

473			326, 286	654

474			321, 285	683

475			326, 286	630

476			322, 286	682

477			318, 286	612

478			318.3	606

479			322, 286	566

480			322, 286	621

481			318, 286	649

482			322, 286	606

483			326, 286	652

484			326, 286	648

485			322, 284	704

486			326, 286	634

487			322, 285	689

488			322, 285	703

489			322	698

490			322, 286	619

491			322, 286	689

Examples 492-621

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-carboxy-2-methoxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be prepared according to method 12 or 14. The corresponding aniline is described in method 22. The amine used to prepare the amide is commercially obtainable, described in method 13, 15, 20, 21, 23, 24 and 25 or in J. Med. Chem. 2003, 46(5), 702-715.

				UV max	MS (ESI)
#	A	R₃′	R₃″	[nm]	(M + H)⁺

492			H	286, 322	584

493			H	286, 322	826

494			H	284, 322	613

495			H	282, 322	640

496			H	286, 320	570

497			H	286, 322	584

498			H	282, 322	693

499			H	286, 322	686

500			H	286, 326	616

501			H	286, 326	630

502			H	282, 325	704

503			H	286, 326	634

504			H	286, 326	648

505			H	286, 322	712

506			H	322, 286	739

507			H	322, 286	645

508			H	326, 286	632

509			H	322, 286	672

510			H	322, 284	700

511			H	314, 286	616

512			H	286, 322	684

513			H	286, 322	670

514			H	282, 322	658

515			H	322, 286	632

516			H	326, 286	628

517			H	325, 286	628

518			H	326, 286	659

519			H	326	699

520			H	284, 326	616

521			H	234, 282, 314	630

522			H	326	660

523			H	326	657

524			H		645

525			H	326	627

526			H	326	660

527			H	326	659

528			H	326	692

529			H	326	644

530			H	326	628

531			H	322	662

532			H	326	699

533			H	326	602

534			H		646

535			H	326	666

536			H	326	646

537			H	326	—

538			H	322	616

539			H	318	630

540			H	318	630

541			H	274	644

542			H	326	658

543			H	286, 324	630

544			H	286, 326	658

545			H	286, 322	630

546			H	286, 326	642

547			H	286, 322	562

548			H	322-326	630

549			H	326	630

550			H	286, 322	607

551			H		646

552			H		644

553			H	326	644

554			H	322-326	658

555			H	322-326	658

556			H	286, 326	658

557			H	322-326	642

558			H	322-326	642

559			H	286, 322	656

560			H	286, 322	656

561			H	286, 322	671

562			H	286, 322	671

563			H	318	685

564			H	322-326	685

565			H	322-326	754

566			H	322-326	672

567			H	322	711

568			H	322-326	711

569			H	326	624

570			H	326	645

571			H	322-326	650

572			H	286, 326	684

573			H	286, 326	684

574			H	326	673

575			H	322	698

576			H	326, 286	646

577			H	286, 322	684

578			H	282, 322	658

579			H	322, 286	617

580			H	326, 286	644

581			H	326, 286	590

582			H	286, 326	673

583			H	326, 285	652

584			H	326, 282	722

585			H	326, 286	648

586			H	326, 285	718

587			H	326, 286	652

588			H	326, 284	652

589			H	325, 283	681

590			H	325.3	652

591			H	326.3	666

592			H	325, 283	666

593			H	325.3	648

594			H	325, 284	648

595			H	325, 284	677

596			H	325, 284	648

597			H	326, 285	662

598			H	325, 284	662

599			H	326, 282	720

600				314, 283	576

601			H	322, 286	714

602			H	286, 322	670

603			H	324, 285	614

604			H	324, 284	684

605			H	324, 285	628

606			H	324, 284	698

607			H	285, 322	630

608			H	325, 284	576

609			H	325, 284	576

610			H	326, 286	659

611			H	326, 286	646

612			H	325, 285	630

613			H	325, 284	630

614			H	325, 285	590

615			H	285, 325	642

616			H	325, 285	670

617			H	326, 286	684

618			H	326, 286	658

619			H	285, 324	684

620			H	326, 286	658

621			H	280, 320	631

Example 622

2-(2-methoxy-4-[2-(2-pyrrolidin-1-yl-ethylcarbamoyl)-ethylamino]-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

73 mg (0.193 mmol) 3-(4-amino-3-methoxy-phenylamino)-N-(2-pyrrolidin-1-yl-ethyl)-propionamide hydrochloride (method 28) are dissolved in 3 ml 2-butanol and combined with 50 mg (0.129 mmol) 2-chloro-4-(2-(2-fluorethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine (method 26). This reaction mixture is stirred for 16 h at 100° C. The solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier material used is C18-RP-silica gel and a gradient is run through which consists at the starting point of 90% water and 10% acetonitrile and at the finishing point of 55% water and 45% acetonitrile. 0.1% formic acid are added both to the water and to the acetonitrile. The suitable fractions are combined with 500 μl of a 1 M aqueous hydrochloric acid and freeze-dried. The product is obtained as the dihydrochloride.
Yield: 33 mg (0.045 mmol; 35%)
UV max: 314 nm
MS (ESI): 659 (M+H)⁺
¹H-NMR: 1.35-1.48 (m, 3H), 1.64-1.78 (m, 4H), 2.37-2.46 (m, 2H), 3.48-3.75 (m, 4H), 3.97-4.14 (m, 1H), 4.50-4.78 (m, 3H), 5.55-5.71 (m, 1H), 6.14-6.42 (m, 2H), 6.96-7.32 (m, 3H), 7.86-7.98 (m, 1H), 8.32 (s, 1H), 8.84 (s, 1H), 10.41 (s, 1H)

Example 623

2-(2-fluoro-ethyl)-7 (2-{4-[4 (2 hydroxy-ethyl)-1H-imidazol-2-yl]-2-methoxy-phenylamine}-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

0.07 g (0.3 mmol) 2-[2-(4-amino-3-methoxy-phenyl)-1H-imidazol-4-yl]-ethanol (method 27) are suspended in 2 ml dioxane and brought into solution in the ultrasound bath at 50° C. 0.8 ml (3.20 mmol) 4 N dioxanic hydrochloric acid are added. The dioxane is eliminated in vacuo, combined with 0.096 g (0.247 mmol) 7-(2-chloro-5-trifluoromethyl-pyrimidine-4-ylamine)-2-(2-fluoro-ethyl)-3-methyl-2,3-dihydro-isoindol-1-one and suspended in butanol. The mixture is stirred for 16 h at 100° C. The crude product is purified by column chromatography. The carrier material used is C18-RP-silica gel. A gradient is run through which consists at the starting point of 75% water and 25% acetonitrile and at the finishing point of 30% water and 70% acetonitrile. 0.1% ammonia is added to the water. 23 mg of this intermediate product and 0.018 g (0.094 mmol) p-toluenesulphonyl chloride are suspended in 0.9 ml of tetrahydrofuran and 0.02 ml (0.139 mmol) triethylamine and combined with 0.007 g (0.057 mmol) 4-dimethylamino-pyridine. This reaction mixture is stirred for 16 h at 20° C. Then it is combined with 0.36 ml (5.064 mmol) pyrrolidine and stirred for 16 h at 60° C. The crude product is purified by column chromatography. The carrier material used is C18-RP-silica gel. A gradient is run through which consists of 90% water and 10% acetonitrile at the starting point and of 60% water and 40% acetonitrile at the finishing point. 0.1% formic acid is added to the water.
Yield: 7 mg (0.011 mmol, 28%)
MS (ESI): 639 (M+H)⁺
UV max: 330 nm
NMR: 1.42-1.46 (m, 3H), 1.78-2.08 (m, 6H), 2.29 (s, 1H), 3.95-4.16 (m, 4H), 4.52-4.78 (m, 3H), 7.09-7.13 (m, 1H), 7.24-7.28 (m, 1H), 7.46-7.50 (m, 1H), 7.52-7.58 (m, 2H), 7.64-7.67 (m, 1H), 7.82-7.88 (m, 1H), 8.02-8.13 (m, 2H), 8.50-8.60 (m, 2H), 9.20-9.23 (m, 1H), 10.52-10.82 (m, 2H).

Examples 624-638

The following compounds are prepared by an analogous process to that described in Example 622 or 623. The corresponding aniline is described in method 27 and 28.

#	B	UV max [nm]	MS (ESI) (M + H)⁺

624		290, 326	586

625		290, 330	654

626		290, 326	625

627		326	512

628		314	685

629		290, 314	659

630			659

631		278	592

632		314	592

633		314	588

634		314	602

635		314	602

636		314	588

637		314	602

638			670

Example 639

2-(4-(4-isopropyl-[1,4]diazepin-1-yl)-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

50 mg (0.087 mmol) 2-(4-(4-[1,4]diazepan-1-yl)-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine (method from Example 622, aniline from method 28) are dissolved in 0.5 ml dimethylacetamide and combined with 13 μl (0.174 mmol) acetone. 37 mg (0.175 mmol) sodium triacetoxyborohydride are added to this reaction mixture. After 16 h at 20° C. the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier material used is C18-RP-silica gel and within 15 min a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid are added both to the water and to the acetonitrile. The suitable fractions are combined with 500 μl of a 1 M aqueous hydrochloric acid and freeze-dried. The product is obtained as the dihydrochloride.
Yield: 51 mg (0.074 mmol; 85%)
UV max: 314 nm
MS (ESI): 616 (M+H)⁺
¹H-NMR: 1.23-1.35 (m, 6H), 1.35-1.51 (m, 3H), 2.16-2.29 (m, 1H), 2.95-3.05 (m, 1H), 3.12-3.23 (m, 1H), 3.42-3.66 (m, 6H), 3.78 (s, 3H), 3.83-4.00 (m, 2H), 4.00-4.16 (m, 1H), 4.50-4.79 (m, 3H), 6.32-6.63 (m, 2H), 7.08-8.59 (m, 4H), 9.24-9.76 (m, 1H), 10.67 (s, 2H)

Examples 640-648

The following compounds are prepared by an analogous process to that described in Example 639.

		UV max	MS (ESI)
#	D	[nm]	(M + H)⁺

640		314	574

641		310-314	628

642		310-314	602

643		310-314	630

644		314	671

645		310-314	618

646		314	658

647		314	588

Examples 648-659

The following compounds are prepared by an analogous process to that described in Example 639. For the reductive amination 2-(2-methoxy-4-piperazin-1-yl-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine is used. The aniline for preparing this compound is described in method 28.

		UV max	MS (ESI)
#	D	[nm]	(M + H)⁺

648		286, 314	631

649		286, 314	603

650		282, 314	643

651		282, 314	671

652		286, 314	657

653		282, 314	628

654		286, 314	657

655		286, 314	671

656		282, 314	614

657		282, 314	560

658		234, 283, 314	694

659		286, 314	574

Examples 660-666

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-Carboxy-2-bromo-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be prepared according to method 29. The corresponding aniline is described in method 22. The amine used to prepare the amide is commercially obtainable or described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

660			314	678/680

661			314	626/628

662			314	626/628

663			286	609/611

664			314	734/736

665			314	693/695

666			286	678/680

Examples 667-681

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-Carboxy-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine may be prepared according to method 14. The corresponding aniline is described in method 22. The amine used to prepare the amide is commercially obtainable or described in method 13. In addition, the group R₃′ may be synthesised analogously to Example 639 by reductive amination. An amine is used which has another protected amino function in the side chain. The protective group used may be a tert-butoxycarbonyl, benzyloxycarbonyl or benzyl group. This protective group is cleaved by a procedure familiar to the skilled man and reductive amination (analogously to Example 639) or alkylation (analogously to method 34 or WO2004052857) are the last steps in this sequence.

				UV max	MS (ESI)
#	A	R₃′	R₃″	[nm]	(M + H)⁺

667			H	314	586

668			H	314	586

669			H	314	586

670			H	314	642

671			H	314	616

672			H	290	600

673			H	290	709

674			H	314	600

675			H	314	586

676				286	574

677			H	286	572

678			H	290	682

679			H	314	642

680			H	290	656

681			H	314	615

Example 682

2-(2-methoxy-4-[3-(4-methyl-piperazin-1-yl)-propionylamino]-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

63 mg (0.116 mmol) 2-(4-acryloylamino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine (method 30) are dissolved in 1 ml of methanol and combined with 70 mg (0.699 mmol) N-methyl-piperazine. After stirring for 48 h at 20° C. the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier material used is C18-RP-silica gel and a gradient is run through within 20 min which consists of 95% water and 5% acetonitrile at the starting point and of 2% water and 98% acetonitrile at the finishing point. 0.1% formic acid are added both to the water and to the acetonitrile. The suitable fractions are combined with 500 μl of a 1 M aqueous hydrochloric acid and freeze-dried. The product is obtained as the dihydrochloride.
Yield: 58 mg (0.081 mmol; 70%)
UV max: 282 nm
MS (ESI): 645 (M+H)⁺
¹H-NMR: 1.42 (d, 3H), 2.18 (s, 3H), 2.29-2.43 (m, 4H), 2.65-2.70 (m, 2H), 3.50-3.62 (m, 1H), 3.72 (s, 3H), 4.00-4.12 (m, 1H), 4.52-4.76 (m, 3H), 7.12-7.17 (m, 1H), 7.12-7.42 (m 4H), 7.51 (s, 1H), 8.17 (s, 1H), 8.38 (s, 1H), 9.08 (s, 1H), 10.18 (s, 1H), 10.46 (s, 1H)

Examples 683-692

The following compounds are prepared by an analogous process to that described in Example 682.

		UV max	MS (ESI)
#	E	[nm]	(M + H)⁺

683		282	661

684		282	673

685		282	701

686		282	645

687		282	685

688		282	616

689		282	713

690		282	630

691		282	632

692		282	602

Examples 693-704

The following compounds are prepared by an analogous process to that described in Example 682. 2-(4-(2-Bromo-acetylamino)-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine or 2-(4-(2-bromo-acetylamino)-2-bromo-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine or 2-[5-(2-bromo-acetylamino)-pyridin-2-ylamino]-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine, which are described in method 30, are used as educt for the nucleophilic substitution.

		UV max	MS (ESI)
#	B	[nm]:	(M + H)⁺:

693		282	685

694		282	685

695		314	659

696		282	645

697		282	644

698		282	618

699		282	602

700		282	687

701		322	573

702		322	630

703		222	650

704		278	707

Example 705

2-(2-methoxy-4-[3-(3-pyrrolidin-1-yl-ethyl)-ureido]-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

70 mg (0.135 mmol) 2-(4-carboxy-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine (analogously to Example 53) are dissolved in 2 ml of toluene and combined with 190 μl (1.348 mmol) triethylamine and 60 μl (0.270 mmol) diphenylphosphorylazide.
This reaction mixture is stirred for 48 h at 20° C. Then the temperature of the suspension is adjusted to 95° C. for 2 h, whereupon a clear brown solution is formed. Then 31 mg (0.270 mmol) 1-(2-aminoethyl)-pyrrolidine are added and the mixture is again stirred for 1 h at 95° C. The solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is C18-RP-silica gel and within 15 min a gradient is run through which consists of 95% water and 5% acetonitrile at the starting point and consists of 2% water and 98% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are made basic with 5 M sodium hydroxide solution and extracted 4 times with 50 ml dichloromethane. The combined organic phases are dried and the solvent is eliminated in vacuo.
Yield: 42 mg (0.067 mmol; 50%)
UV max: 282 nm
MS (ESI): 631 (M+H)⁺
¹H-NMR: 1.42-1.48 (m, 3H), 1.69-1.79 (m, 4H), 3.22-3.28 (m, 2H), 3.49-3.62 (m, 1H), 3.70 (s, 3H), 3.99-4.12 (m, 1H), 4.53-4.76 (m, 3H), 6.17 (s, 1H), 6.84-6.91 (m, 1H), 7.15-7.33 (m, 3H), 7.40 (s, 1H), 8.36 (s, 1H), 8.76 (s, 1H), 9.01 (s, 1H), 10.44 (s, 1H)

Example 706

2-(2-methoxy-4-ureido-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

This compound is prepared analogously to Example 705.
UV max: 282/314 nm
MS (ESI): 534 (M+H)⁺
¹H-NMR: 1.42 (d, 3H), 3.48-3.64 (m, 1H), 3.69 (s, 3H), 3.98-4.13 (m, 1H), 4.50-4.77 (m, 3H), 5.89 (s, 2H), 6.94 (d, 1H), 7.16-7.30 (m, 2H), 7.36 (s, 1H), 8.33-8.41 (m, 2H), 8.38 (s, 1H), 8.73 (s, 1H), 9.00 (s, 1H), 10.44 (s, 1H)

Example 707

2-(2-methoxy-4-[(1-methyl-piperidin-4-carbonyl)-amino]-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

Starting from 2-(4-amino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine (method 30) the above-mentioned product is prepared using an amide linking method familiar to the skilled man (cf. also Example 53 or 1032). The substance is obtained as a free base.
UV max: 282 nm
MS (ESI): 616 (M+H)⁺
¹H-NMR (400 MHz, CDCl₃): 1.51 (d, 3H), 2.25-2.32 (m, 1H), 2.36 (s, 3H), 3.00-3.07 (m, 2H), 3.53-3.65 (m, 1H), 3.92 (s, 3H), 4.13-4.27 (m, 1H), 4.56-4.77 (m, 3H), 6.84 (d, 1H), 7.07 (d, 1H), 7.44 (s, 1H), 7.47-7.54 (m, 1H), 7.57 (s, 1H), 7.62 (s, 1H), 8.16-8.24 (m, 1H), 8.39 (s, 1H), 8.60-8.68 (m, 1H), 10.42 (s, 1H)

Examples 708-795

Using an analogous method to that described in Example 53 a primary amine which has another protected amino function in the side chain is coupled to 2-(4-carboxy-2-methoxy-phenylamino)-4-[2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino]-5-trifluoromethyl-pyrimidine. The protective group used may be a tert-butoxycarbonyl, benzyloxycarbonyl or benzyl group. This protective group is cleaved using a procedure familiar to the skilled man and reductive amination (analogously to Example 639) or alkylation (analogously to method 34 or WO2004052857) are the final steps in this sequence.

			MS
		UV	(ESI)
		max	(M +
#	R₃′	[nm]	H)⁺

708		285, 322	706

709		285, 322	656

710		285, 322	630

711		322, 286	644

712		325, 286	699

713		282, 318	644

714		326	685

715		326	658

716		326	699

717		326	630

718		326	644

719		322	644

720		326	656

721		326	678

722		314	630

723		322	641

724		326	712

725		326	642

726		322	642

727		318	672

728		301	686

729		326	588

730		326	642

731		326	670

732		326	642

733		326	630

734		326	699

735		310	616

736		326	656

737		322	630

738		326	656

739		326	656

740		266	652

741		326	629

742		326	671

743		326	630

744		326	642

745		326	602

746		326	628

747		326	616

748		326	602

749		322	652

750		326	646

751		326	672

752		326	616

753		326	616

754		326	685

755		322	616

756		318	713

757		286, 322	588

758		226, 286, 322	602

759		322- 326	656

760		322- 326	699

761		322- 326	670

762		322- 326	699

763		322	713

764		326	685

765		322	684

766		326	642

767		322- 326	656

768		322- 326	685

769		322- 326	630

770		286, 322	670

771		286, 322	670

772		322- 326	644

773		322	684

774		322- 326	658

775		322	686

776		322- 326	727

777		322- 326	674

778		322- 326	684

796		322- 326	698

780		286, 322	630

781		282, 314	616

782		322, 286	686

783		326	684

784		324, 286	656

785		326, 286	685

786		322, 286	715

787		322, 286	673

788		285, 322	616

789		285, 322	630

790		285, 322	686

791		285, 322	686

792		326	644

793		322	630

794		326	631

795		326	660

Example 796

2-[2-methoxy-4-(2-methyl-2-pyrrolidin-1-yl-propylcarbamoyl)-phenylamino]-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

200 mg (0.385 mmol) 2-(4-carboxy-2-methoxy-phenylamino)-4-[2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino]-5-trifluoromethyl-pyrimidine (analogously to Example 53) are dissolved in 1 ml of dimethylformamide cooled to 0° C. and combined with 520 μl (3.038 mmol) diisopropylethylamine and 160 mg (0.498 mmol) O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-tetrafluoroborate. This solution is slowly added dropwise after 10 min to 56 μl (0.539 mmol) 1,2-diamino-2-methylpropane, which is dissolved in 300 μl dimethylformamide. The reaction mixture is stirred for 24 h at 20° C. and then the solvent is eliminated in vacuo. The residue is purified by column chromatography. The carrier used is C18-RP-silica gel and within 15 min a gradient is run through which consists at the starting point of 90% water and 10% acetonitrile and at the finishing point of 50% water and 50% acetonitrile. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are freeze-dried. This intermediate product is combined with 70 mg (0.515 mmol) potassium carbonate and with 84 mg (0.506 mmol) potassium iodide and suspended in 2 ml acetonitrile. 20 μl (0.170 mmol)1,4-dibromobutane are added to this mixture and it is stirred under reflux conditions for 16 h. Then the solvents are solvent eliminated in vacuo and the residue is purified by column chromatography. The carrier used is C18-RP-silica gel and within 15 min a gradient is run through which consists at the starting point of 90% water and 10% acetonitrile and at the finishing point of 50% water and 50% acetonitrile. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are combined with 0.5 ml 1 N hydrochloric acid and freeze-dried. The product is obtained as the dihydrochloride.
Yield: 20 mg (0.032 mmol, 8%)
UV max: 325 nm
MS (ESI): 644 (M+H)⁺
¹H-NMR (400 MHz): 1.30-1.47 (m, 9H), 1.85-2.01 (m, 4H), 3.20-3.31 (m, 2H), 3.91 (s, 3H), 3.99-4.15 (m, 1H), 4.51-4.78 (m, 3H), 7.23-7.29 (m, 1H), 7.39-7.47 (m, 1H), 7.63-7.69 (m, 1H), 7.73-7.77 (m, 1H), 7.79-7.87 (m, 1H), 8.40-8.59 (m, 2H), 8.75-8.82 (m, 1H), 9.16-9.21 (m, 1H), 10.50-10.63 (m, 2H)

Examples 797-806

The following compounds are prepared by an analogous method to that described in Example 796:

		UV max	MS (ESI)
#	R₃′	[nm]	(M + H)⁺

797	285, 325	642

798	284, 325	642

799	325, 285	644

800	325, 285	644

801	325, 285	644

802	325, 285	656

803	325, 285	658

804	325, 284	658

805	326, 286	670

806	324, 285	670

Examples 807-821

The following compounds are prepared by an analogous process to that described in Example 53. The corresponding aniline is described in method 31. The amine used to prepare the amide is commercially obtainable or is described in method 13, 21 or in method 25.

		UV max	MS (ESI)
#	R₃′	[nm]	(M + H)⁺

807		286, 322	686

808		286, 322	616

809		286, 322	630

810		286, 322	616

811		286, 322	712

812		322, 286	684

813			689

814		278	689

815		322	630

816		286, 326	645

817		285, 322	659

818		285, 322	616

819		285, 322	630

820			630

821		322, 286	630

Examples 822-885

The following compounds are prepared by an analogous process to that described in Example 53. The corresponding aniline is described in method 31. The amine used to prepare the amide is commercially obtainable, described in method 13, 15, 20, 21, 23, 24 and 25 or in J. Med. Chem. 2003, 46(5), 702-715.

		UV max	MS (ESI)
#	R₃′	[nm]	(M + H)⁺

822		286, 322	686

823		325, 284	616

824		286, 326	630

825		286, 322	616

826		286, 318	712

827		286, 322	684

828		326	645

829		316	689

830		322	689

831			616

832		318	630

833		326	588

834		322	630

835		286, 322	630

836			658

837		322-326	602

838		322-326	616

839		322	616

840		322-326	616

841		322-326	630

842		322-326	630

843		286, 322	644

844		286, 322	642

845		286, 322	642

846		286, 322	656

847		282, 318	630

848		282, 322	630

849		286, 318	671

850		286, 322	630

851		286, 322	630

852		286, 322	644

853		322-326	672

854		322	672

855		286, 322	725

856		286, 322	725

857		322-326	685

858		286, 322	713

859		286, 322	713

860		286, 322	644

861		286, 322	644

862		318-322	645

863		286, 322	658

864		286, 322	699

865		286, 322	699

866		326	709

867		322	697

868		322	697

869		318	695

870		290.3	693

871		322	695

872		286, 322	753

873		286, 326	642

874		286, 322	645

875		322, 286	659

876		282, 322	684

877		324, 284	646

878		286, 322	670

879		325, 284	630

880		322, 286	630

881		322, 286	684

882		325, 286	670

883		322, 286	646

884		326, 286	644

885		325, 285	630

Examples 886-891

The following compounds are prepared by an analogous process to that described in Example 622 or 623. The corresponding aniline is described in method 27 or 28.

		UV max	MS (ESI)
#	B	[nm]	(M + H)⁺

886		314	685

887		314	685

888		286, 310	685

889		282, 314	699

890		338	656

891		314	588

Examples 892-894

The following compounds are prepared by an analogous process to that described in Example 53. 2-(4-carboxy-2-bromo-phenylamino)-4-chloro-5-trifluoromethyl-pyrimidine is described in method 29. The corresponding aniline is described in method 31. The amine used to prepare the amide is commercially obtainable.

		UV max	MS (ESI)
#	R₃′	[nm]	(M + H)⁺

892		314	665

893		270	665

894		270	680

Example 895

2-(2-methoxy-4-[(1-methyl-piperidin-4-carbonyl)-amino]-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine Enantiomer 1

Starting from 2-(4-amino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine enantiomer 1 (analogously to method 30) the above-mentioned product is prepared by an amide linking method familiar to the skilled man (cf. also Example 1032). It is obtained as the dihydrochloride.
UV max: 310 nm
MS (ESI): 616 (M+H)⁺
¹H-NMR (500 MHz): 1.42 (d, 3H), 1.69-1.77 (m, 2H), 1.77-1.84 (m, 2H), 1.94-2.03 (m, 2H), 2.23 (s, 3H), 2.29-2.38 (m, 1H), 2.86-2.93 (m, 2H), 3.72 (s, 3H), 4.00-4.12 (m, 1H), 4.52-4.75 (m, 3H), 7.16 (d, 3H), 7.18-7.24 (m, 1H), 7.32-7.41 (m, 1H), 7.57 (s, 1H), 8.18 (s, 1H), 8.38 (s, 1H), 9.07 (s, 1H), 9.95 (s, 1H), 10.46 (s, 1H)

Example 896

2-(2-methoxy-4-(2-pyrrolidin-1-yl-acetylamino)-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

Enantiomer 1

Starting from 2-(4-amino-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine Enantiomer 1 (analogously to method 30) the above-mentioned product is prepared by an amide linking method familiar to the skilled man (cf. also Example 1032). It is obtained as the dihydrochloride.
UV max: 282 nm
MS (ESI): 602 (M+H)⁺
¹H-NMR (500 MHz): 1.43 (d, 3H), 1.87-2.00 (m, 2H), 2.00-2.10 (m, 2H), 3.12-3.22 (m, 2H), 3.74 (s, 3H), 4.00-4.13 (m, 1H), 4.28-4.32 (m, 2H), 4.53-4.76 (m, 3H), 7.19-7.49 (m, 4H), 7.51 (s, 1H), 8.41 (s, 1H), 9.26 (s, 1H), 10.20-10.31 (m, 1H), 10.54 (s, 1H), 10.86 (s, 1H)

Examples 897-952

Using a method analogous to that described in Example 53 a primary amine which has another protected amino function in the side chain is coupled to 2-(4-carboxy-2-methoxy-phenylamino)-4-[2-(2-fluoro-ethyl)-1-methyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino]-5-trifluoromethyl-pyrimidine Enantiomer 1. The protective group used may be a tert-butoxycarbonyl, benzyloxycarbonyl or benzyl group. This protective group is cleaved using a procedure familiar to the skilled man and reductive amination (analogously to Example 639) or alkylation (analogously to method 34 or WO2004052857) are the final steps in this sequence.

		UV max	MS (ESI)
#	R₃′	[nm]	(M + H)⁺

897			672

898		322	644

899		326	630

900		326	630

901		322	644

902		322	642

903		322	658

904		326	615

905		322	656

906		326	658

907		326	644

908		322	644

909		322	670

910		306	686

911		326	630

912			666

913		286, 322	656

914		286, 322	656

915		286, 318	670

916		286, 322	713

917		286, 322	670

918		286.3	713

919		286, 322	642

920		286, 322	672

921		286, 322	672

922		286, 322	644

923		286, 322	670

924		286, 322	700

925		286, 322	700

926		286, 322	670

927		326	713

928		322-326	700

929		322-326	644

930		322	658

931		322-326	713

932		322	700

933		322-326	644

934		322	658

935		322-326	714

936		322	714

937		322	662

938		322-326	662

939			676

940		322-326	680

941		286, 322	648

942		230, 286, 318	662

943		384, 324	668

944		282, 322	670

945		282, 322	696

946		228, 284, 322	642

947		226, 286, 322	672

948		286, 322	644

949		324, 284	644

950		285, 322	616

951		285, 325	630

952		285, 325	616

Examples 953-958

The following compounds are prepared by a method analogous to that described in Example 796:

		UV max	MS (ESI)
#	R₃′	[nm]	(M + H)⁺

953		326, 286	658

954		325, 285	670

955		325, 285	670

956		325, 284	644

957		325, 284	658

958		325, 285	672

Example 959

2-(2-methoxy-4-(2-pyrrolidin-1-yl-ethylcarbamoyl)-phenylamino)-4-(2-(2-fluoro-ethyl)-1-ethyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

The racemic synthesis of the above-mentioned compound is carried out using by a method analogous to that described in Example 53. The corresponding aniline is described in method 22. The two enantiomers are isolated by preparative chromatography:
column: 250×4.6 mm CHIRALPAKADH®
eluant: 25 ethanol/75 methanol (v/v) (0.03% triethylamine is added to each solvent)
flow rate: 0.5 ml/min
temperature: 20° C.
The enantiomer that elutes first is referred to as Enantiomer 1 and bears the symbol *1 in the chemical formula.

Enantiomer 1

retention time: 9.96 min
The enantiomer that elutes second is referred to as Enantiomer 2 and bears the symbol *2 in the chemical formula.

Enantiomer 2

retention time: 12.60 min

Examples 960-976

The following compounds are prepared by an analogous method to that described in Example 53. The corresponding aniline is described in method 22. The amine used to prepare the amide is commercially obtainable or is described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

960			280, 320	654

961			282, 318

962			286, 322	680

963			286, 326	630

964			286, 326	644

965			286, 326	630

966			286, 326	659

967			286, 326	630

968			286, 322	644

969			286, 326	644

970			286, 326	644

971			286, 326	714

972			286, 322	632

973			286, 326	646

974			286, 326	660

975			282, 326	685

976			282, 326	659

Examples 977-980

The following compounds are prepared by an analogous method to that described in Example 53. The corresponding aniline is described in method 6. The amine used to prepare the amide is described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

977			234, 282, 318	655

978			226, 282, 318	655

979			222, 282, 318	641

980			230, 282, 314	671

Examples 981-999

The following compounds are prepared by an analogous method to that described in Example 53. The corresponding aniline is described in method 32. The amine used to prepare the amide is commercially obtainable or described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

981			318	612
982			318	583
983			322	599

984				639

985			286	706

986			322	597

987			318	679

988			286	653

989			322	611

990			322	583

991			318	625

992			318	597

993			318	598

994			318	569

995			322	585

996			286	639

997			318	626

998			318	599

999			318	318

Examples 1000-1024

The following compounds are prepared by an analogous method to that described in Example 53. The corresponding aniline is described in method 33. The amine used to prepare the amide is commercially obtainable or described in method 13 or 21.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

1000			282, 322	614

1001			282, 322	841

1002			282, 326	571

1003			280, 322	655

1004			280, 325	655

1005			280, 322	669

1006			280, 325	599

1007			282, 327	613

1008			280, 322	697

1009			282, 325	627

1010			283, 328	641

1011			280, 325	585

1012			280, 325	599

1013			326, 283	585

1014			282, 327	599

1015			322-326	597

1016			326	611

1017			280, 325	585

1018			280, 325	614

1019			280, 325	585

1020			280, 322	599

1021			280, 325	641

1022			280, 325	599

1023			280, 325	585

1024			280, 322	663

Examples 1025-1032

The following compounds are prepared by an analogous method to that described in Example 53. The corresponding aniline is described in method 10. The amine used to prepare the amide is commercially obtainable or described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

1025			318	648

1026			318	359

1027			322	662

1028			322	662

1029			322	664

1030			226, 318	678

1031			226, 318	691

1032			322	648

Examples 1033-1035

The following compounds are prepared by an analogous method to that described in Example 53. The corresponding aniline is described in method 2. The amine used to prepare the amide is described in method 13.

		MS (ESI)
#	R₃′	(M + H)⁺	salt form

1033		701	base

1034		645	formate

1035		631	formate

Example 1036

(2-methoxy-4-(2-pyrrolidin-1-yl-ethylcarbamoyl)-phenylamino)-4-(2-(2-fluoro-ethyl)-1,1-dimethyl-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-trifluoromethyl-pyrimidine

The above-mentioned compound is prepared by a method analogous to that described in Example 53. The corresponding aniline is described in method 34. The amine used to prepare the amide is commercially obtainable. The substance is obtained as the dihydrochloride.
UV max: 326, 286 nm
MS (ESI): 630 (M+H)⁺
¹H-NMR (400 MHz): 1.44-1.50 (m, 6H), 1.84-1.95 (m, 2H), 1.98-2.07 (m, 2H), 3.02-3.12 (m, 2H), 3.62-3.70 (m, 4H), 3.71-3.76 (m, 1H), 3.77-3.81 (m, 1H), 3.89 (s, 3H), 4.57-4.61 (m, 1H), 4.69-4.73 (m, 1H), 7.27-7.31 (m, 1H), 7.39-7.45 (m, 1H), 7.55-7.59 (m, 1H), 7.63-7.66 (m, 1H), 7.84-7.88 (m, 1H), 8.44-8.55 (m, 2H), 8.77-8.82 (m, 1H), 9.11-9.15 (m, 1H), 9.91-10.03 (m, 1H), 10.51-10.55 (m, 1H)

Example 1037

2-(2-methoxy-4-[2-(4-methyl-piperazin-1-yl)-ethylcarbamoyl]-phenylamino)-4-(2-(2-fluoro-ethyl)-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-acetyl-pyrimidine

50 mg (0.104 mmol) 2-(4-carboxy-2-methoxy-phenylamino)-4-(2-(2-fluoro-ethyl)-3-oxo-2,3-dihydro-1H-isoindol-4-ylamino)-5-acetyl-pyrimidine (prepared by an analogous process to that described in Example 622 or 623) are dissolved in 0.5 ml of dimethylformamide and combined with 72 μl (0.520 mmol) and 34 mg (0.104 mmol) O-(benzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium-tetrafluoroborate. After stirring for 20 min at 20° C., 23 mg (0.156 mmol) 2-(4-methylpiperazin-1-yl)-ethylamine are added. The reaction is completed after 2 h at 20° C. Then the solvent is eliminated in vacuo and the residue is purified by column chromatography. The carrier used is C18-RP-silica gel and a gradient is run through within 20 min which consists of 95% water and 5% acetonitrile at the starting point and consists of 5% water and 95% acetonitrile at the finishing point. 0.1% formic acid are added to both the water and to the acetonitrile. The suitable fractions are combined with 500 μl of a 1 M aqueous hydrochloric acid and freeze-dried. The product is obtained as the trihydrochloride.
UV max: 326 nm
MS (ESI): 605 (M+H)⁺
¹H-NMR (500 MHz): 2.53-2.58 (m, 3H), 2.80-2.92 (m, 3H), 3.62-3.88 (m, 9H), 3.88-4.01 (m, 4H), 4.54 (s, 2H), 4.58-4.66 (m, 1H), 4.69-4.77 (m, 1H), 7.14-7.32 (m, 1H), 7.32-7.50 (m, 1H), 7.50-7.59 (m, 1H), 7.63-7.75 (m, 1H), 7.78-8.01 (m, 1H), 8.29-8.60 (m, 1H), 8.73-8.99 (m, 2H), 9.03-9.18 (m, 1H), 12.31-12.41 (m, 1H)

Examples 1038-1060

The following compounds are prepared by an analogous method to that described in Example 1037. The aniline used is described in method 28.
The amine used to prepare the amide is commercially obtainable or described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

1038			326	660

1039			326	646

1040			328	576

1041			318	672

1042			326	605

1043			330	590

1044			318	663

1045			330	604

1046			326	686

1047			326	604

1048			330	590

1049			326	713

1050			330	590

1051			250	614

1052			334-338	600

1053			334-338	614

1054			338	600

1055			338	670

1056			334	696

1057			330	622

1058			327	340

1059			330	608

1060			330	632

Examples 1061-1069

The following compounds are prepared by an analogous method to that described in Example 622 or 623. The corresponding aniline is described in method 28.

			UV max	MS (ESI)
#	A	B	[nm]	(M + H)⁺

1061			254, 316	552

1062			254, 314	548

1063			250	598

1064			254, 318	588

1065			250	518

1066			252, 318	606

1067			250, 310	566

1068			254, 318	552

1069			262; 314- 318	566

Examples 1070-1071

The following compounds are prepared by an analogous method to that described in Example 622 or 623 and 53. The corresponding aniline is described in method 28. The amine used to prepare the amide is commercially obtainable or described in method 13.

			UV max	MS (ESI)
#	A	R₃′	[nm]	(M + H)⁺

1070			330	608

1071			330	678

Examples 1072-1085

The following compounds are prepared by an analogous method to that described in Example 1037. The corresponding aniline is described in method 28. The amine used to prepare the amide is commercially obtainable or described in method 13.

			UV max	MS (ESI)
#	Z	R₃′	[nm]	(M + H)⁺


1072			285, 320	674

1073			326	663

1074			306	596

1075			326	593

1076			262	596

1077			326	593

1078			318	652

1079			325	582

1080			319	582

1081			302	666

1082			322	626

1083			318	626

1084			286, 318	612

1085			280, 325	572

Biological Properties

As demonstrated by DNA staining followed by FACS analysis, the inhibition of proliferation brought about by the compounds according to the invention is mediated above all by the arrest of the cells in the G2/M phase of the cell cycle. The cells arrest, depending on the type of cell used, for a specific length of time in this cell cycle phase before programmed cell death is initiated. An arrest in the G2/M phase of the cell cycle may be initiated e.g. by the inhibition of specific cell cycle kinases. On the basis of their biological properties the compounds of general formula I according to the invention, their isomers and the physiologically acceptable salts thereof are suitable for treating diseases characterised by excessive or anomalous 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; leukaemias, lymphomas and solid tumours; skin diseases (e.g. psoriasis); bone diseases; cardiovascular diseases (e.g. restenosis and hypertrophy). They are also useful for protecting proliferating cells (e.g. hair, intestinal, blood and progenitor cells) from DNA damage caused by radiation, UV treatment and/or cytostatic treatment (Davis et al., 2001). The new compounds may be used for the prevention, short- or long-term treatment of the above-mentioned diseases, also in combination with other active substances used for the same indications, e.g. cytostatics, steroids or antibodies.
The activity of the compounds according to the invention on various kinases, for example on serine-threonine kinase PLK-1, was determined by in vitro kinase assays with recombinantly produced protein. In this assay the compounds exhibit a good to very good effect on PLK1, i.e. for example an IC50 value of less than 1 μmol/L, usually less than 0.1 μmol/L.

Example PLK-1 Kinase Assay

Recombinant human PLK1 enzyme linked to GST at its N-terminal end is isolated from insect cells infected with baculovirus (Sf21). Purification is carried out by affinity chromatography on glutathione sepharose columns.
4×10⁷Sf21 cells (Spodoptera frugiperda) in 200 ml of Sf-900 II Serum free insect cell medium (Life Technologies) are seeded in a spinner flask. After 72 hours' incubation at 27° C. and 70 rpm, 1×10⁸Sf21 cells are seeded in a total of 180 ml medium in a new spinner flask. After another 24 hours, 20 ml of recombinant Baculovirus stock suspension are added and the cells are cultivated for 72 hours at 27° C. at 70 rpm. 3 hours before harvesting, okadaic acid is added (Calbiochem, final concentration 0.1 μM) and the suspension is incubated further. The cell number is determined, the cells are removed by centrifuging (5 minutes, 4° C., 800 rpm) and washed 1× with PBS (8 g NaCl/l, 0.2 g KCl/l, 1.44 g Na₂HPO₄/l, 0.24 g KH₂PO4/l). After centrifuging again the pellet is flash-frozen in liquid nitrogen. Then the pellet is quickly thawed and resuspended in ice-cold lysing buffer (50 mM HEPES pH 7.5, 10 mM MgCl₂, 1 mM DTT, 5 μg/ml leupeptin, 5 μg/ml aprotinin, 100 μM NaF, 100 μM PMSF, 10 mM β-glycerolphosphate, 0.1 mM Na₃VO₄, 30 mM 4-nitrophenylphosphate) to give 1×10⁸cells/17.5 ml. The cells are lysed for 30 minutes on ice. After removal of the cell debris by centrifugation (4000 rpm, 5 minutes) the clear supernatant is combined with glutathione sepharose beads (1 ml resuspended and washed beads per 50 ml of supernatant) and the mixture is incubated for 30 minutes at 4° C. on a rotating board. Then the beads are washed with lysing buffer and the recombinant protein is eluted from the beads with 1 ml eluting buffer/ml resuspended beads (eluting buffer: 100 mM Tris/HCl pH=8.0, 120 mM NaCl, 20 mM reduced glutathione (Sigma G-4251), 10 mM MgCl₂, 1 mM DTT). The protein concentration is determined by Bradford Assay.

Assay

The following components are combined in a well of a 96-well round-bottomed dish (Greiner bio-one, PS Microtitre plate No. 650101):

- 10 μl of the compound to be tested in variable concentrations (e.g. beginning at 300 μM, and dilution to 1:3) in 6% DMSO, 0.5 mg/ml casein (Sigma C-5890), 60 mM β-glycerophosphate, 25 mM MOPS pH=7.0, 5 mM EGTA, 15 mM MgCl₂, 1 mM DTT
- 20 μl substrate solution (25 mM MOPS pH=7.0, 15 mM MgCl₂, 1 mM DTT, 2.5 mM EGTA, 30 mM β-glycerophosphate, 0.25 mg/ml casein)
- 20 μl enzyme dilution (1:100 dilution of the enzyme stock in 25 mM MOPS pH=7.0, 15 mM MgCl₂, 1 mM DTT)
- 10 μl ATP solution (45 μM ATP with 1.11×10⁶Bq/ml gamma-P33-ATP).

The reaction is started by adding the ATP solution and continued for 45 minutes at 30° C. with gentle shaking (650 rpm on an IKA Schüttler MTS2). The reaction is stopped by the addition of 125 μl of ice-cold 5% TCA per well and incubated on ice for at least 30 minutes. The precipitate is transferred by harvesting onto filter plates (96-well microtitre filter plate: UniFilter-96, GF/B; Packard; No. 6005177), then washed four times with 1% TCA and dried at 60° C. After the addition of 35 μl scintillation solution (Ready-Safe; Beckmann) per well the plate is sealed shut with sealing tape and the amount of P33 precipitated is measured with the Wallac Betacounter. The measured data are evaluated using the standard Graphpad software (Levenburg-Marquard Algorhythmus).
The anti-proliferative activity of the compounds according to the invention is determined in the cytotoxicity test on cultivated human tumour cells and/or in a FACS analysis, for example on HeLa S3 cells. In both test methods the compounds exhibit good to very good activity, i.e. for example an EC50 value in the HeLa S3 cytotoxicity test of less than 5 mmol/L, generally less than 1 μmol/L.

Measurement of Cytotoxicity on Cultivated Human Tumour Cells

To measure cytotoxicity on cultivated human tumour cells, cells of cervical carcinoma tumour cell line HeLa S3 (obtained from American Type Culture Collection (ATCC)) are cultivated in Ham's F12 Medium (Life Technologies) and 10% foetal calf serum (Life Technologies) and harvested in the log growth phase. Then the HeLa S3 cells are placed in 96-well plates (Costar) at a density of 1000 cells per well and incubated overnight in an incubator (at 37° C. and 5% CO2), while on each plate 6 wells are filled with medium alone (3 wells as the medium control, 3 wells for incubation with reduced AlamarBlue reagent). The active substances are added to the cells in various concentrations (dissolved in DMSO; DMSO final concentration: 0.1%) (in each case as a triple measurement). After 72 hours incubation 20 μl AlamarBlue reagent (AccuMed International) are added to each well, and the cells are incubated for a further 5-7 hours. As a control, 20 μl reduced AlamarBlue reagent is added to each of 3 wells (AlamarBlue reagent, which is autoclaved for 30 min) After incubation the colour change of the AlamarBlue reagent in the individual wells is determined in a Perkin Elmer fluorescence spectrophotometer (excitation 530 nm, emission 590 nm, slits 15, integrate time 0.1). The amount of AlamarBlue reagent reacted represents the metabolic activity of the cells. The relative cell activity is calculated as a percentage of the control (HeLa S3 cells without inhibitor) and the active substance concentration which inhibits the cell activity by 50% (IC50) is derived. The values are calculated from the average of three individual measurements—with correction of the dummy value (medium control).

FACS Analysis

Propidium iodide (PI) binds stoichiometrically to double-stranded DNA, and is thus suitable for determining the proportion of cells in the G1, S, and G2/M phase of the cell cycle on the basis of the cellular DNA content. Cells in the G0 and G1 phase have a diploid DNA content (2N), whereas cells in the G2 or mitosis phase have a 4N DNA content.
For PI staining, for example, 1×10⁶HeLa S3 cells are seeded onto a 75 cm2 cell culture flask, and after 24 h either 0.1% DMSO is added as control or the substance is added in various concentrations (in 0.1% DMSO). The cells are incubated for 24 h with the substance or with DMSO before the cells are washed 2× with PBS and then detached with trypsin/EDTA. The cells are centrifuged (1000 rpm, 5 min, 4° C.), and the cell pellet is washed 2× with PBS before the cells are resuspended in 0.1 ml PBS. Then the cells are fixed with 80% ethanol for 16 hours at 4° C. or alternatively for 2 hours at −20° C. The fixed cells are centrifuged (1000 rpm, 5 min, 4° C.), washed with PBS and then centrifuged again. The cell pellet is resuspended in 2 ml 0.25% Triton X-100 in PBS, and incubated on ice for 5 min before 5 ml PBS are added and the mixture is centrifuged again. The cell pellet is resuspended in 350 μl PI staining solution (0.1 mg/ml RNase A (Sigma, No. R-4875), 10 μg/ml prodium iodide (Sigma, No. P-4864) in 1×PBS). The cells are incubated for 20 min in the dark with the staining buffer before being transferred into sample measuring containers for the FACS scan. The DNA measurement is carried out in a Becton Dickinson FACS Analyzer, with an argon laser (500 mW, emission 488 nm), and the DNA Cell Quest Programme (BD). The logarithmic PI fluorescence is determined with a band-pass filter (BP 585/42). The cell populations in the individual cell cycle phases are quantified using the ModFit LT Programme made by Becton Dickinson.
The compounds according to the invention are also tested accordingly for other tumour cells. For example, these compounds are effective on carcinomas of all kinds of tissue (e.g. breast (MCF7); colon (HCT116), head and neck (FaDu), lung (NCI-H460), pancreas (BxPC-3), prostate (DU145)), sarcomas (e.g. SK-UT-1B), leukaemias and lymphomas (e.g. HL-60; Jurkat, THP-1) and other tumours (e.g. melanomas (BRO), gliomas (U-87MG)) and could be used for such indications. This is evidence of the broad applicability of the compounds according to the invention for the treatment of all kinds of tumour types.
The compounds of general formula (I) may be used on their own or in conjunction with other active substances according to the invention, optionally also in conjunction with other pharmacologically active substances.
Suitable preparations include for example tablets, capsules, suppositories, solutions, particularly solutions for injection (s.c., i.v., i.m.) and infusion, elixirs, emulsions or dispersible powders. The content of the pharmaceutically active compound(s) should be in the range from 0.1 to 90 wt.-%, preferably 0.5 to 50 wt.-% of the composition as a whole, i.e. in amounts which are sufficient to achieve the dosage range specified below. The doses specified may, if necessary, be given several times a day.
Suitable tablets may be obtained, for example, by mixing the active substance(s) 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 gelatine, lubricants such as magnesium stearate or talc and/or agents for delaying release, such as carboxymethyl cellulose, cellulose acetate phthalate, or polyvinyl acetate. The tablets may also comprise several layers.
Coated tablets may be prepared accordingly by coating cores produced analogously to the tablets with substances normally used for tablet coatings, for example collidone or shellac, gum arabic, talc, titanium dioxide or sugar. To achieve delayed release or prevent incompatibilities the core may also consist of a number of layers. Similarly the tablet coating may consist of a number or layers to achieve delayed release, possibly using the excipients mentioned above for the tablets.
Syrups or elixirs containing the active substances or combinations thereof according to the invention may additionally contain a sweetener such as saccharine, cyclamate, glycerol or sugar and a flavour enhancer, e.g. a flavouring such as vanillin or orange extract. They may also contain suspension adjuvants or thickeners such as sodium carboxymethyl cellulose, wetting agents such as, for example, condensation products of fatty alcohols with ethylene oxide, or preservatives such as p-hydroxybenzoates.
Solutions for injection and infusion are prepared in the usual way, e.g. with the addition of isotonic agents, preservatives such as p-hydroxybenzoates, or stabilisers such as alkali metal salts of ethylenediamine tetraacetic acid, optionally using emulsifiers and/or dispersants, whilst if water is used as the diluent, for example, organic solvents may optionally be used as solvating agents or dissolving aids, and transferred into injection vials or ampoules or infusion bottles.
Capsules containing one or more active substances or combinations of active substances may for example be prepared by mixing the active substances with inert carriers such as lactose or sorbitol and packing them into gelatine capsules.
Suitable suppositories may be made for example by mixing with carriers provided for this purpose, such as neutral fats or polyethyleneglycol or the derivatives thereof. Excipients which may be used include, for example, water, pharmaceutically acceptable organic solvents such as paraffins (e.g. petroleum fractions), vegetable oils (e.g. groundnut or sesame oil), mono- or polyfunctional alcohols (e.g. ethanol or glycerol), carriers such as e.g. natural mineral powders (e.g. kaolins, clays, talc, chalk), synthetic mineral powders (e.g. highly dispersed silicic acid and silicates), sugars (e.g. cane sugar, lactose and glucose) emulsifiers (e.g. lignin, spent sulphite liquors, methylcellulose, starch and polyvinylpyrrolidone) and lubricants (e.g. magnesium stearate, talc, stearic acid and sodium lauryl sulphate).
The preparations are administered by the usual methods, preferably by oral or transdermal route, most preferably by oral route. For oral administration the tablets may, of course contain, apart from the abovementioned carriers, additives such as sodium citrate, calcium carbonate and dicalcium phosphate together with various additives such as starch, preferably potato starch, gelatine and the like. Moreover, lubricants such as magnesium stearate, sodium lauryl sulphate and talc may be used at the same time for the tabletting process. In the case of aqueous suspensions the active substances may be combined with various flavour enhancers or colourings in addition to the excipients mentioned above.
For parenteral use, solutions of the active substances with suitable liquid carriers may be used.
The dosage for intravenous use is from 1-1000 mg per hour, preferably between 5 and 500 mg per hour.
However, it may sometimes be necessary to depart from the amounts specified, depending on the body weight, the route of administration, the individual response to the drug, the nature of its formulation and the time or interval over which the drug is administered. Thus, in some cases it may be sufficient to use less than the minimum dose given above, whereas in other cases the upper limit may have to be exceeded. When administering large amounts it may be advisable to divide them up into a number of smaller doses spread over the day.
The formulation examples which follow illustrate the present invention without restricting its scope:

Examples of Pharmaceutical Formulations


A)	Tablets	per tablet

active substance	100	mg
lactose	140	mg
corn starch	240	mg
polyvinylpyrrolidone	15	mg
magnesium stearate	5	mg
	500	mg

The finely ground active substance, lactose and some of the corn starch are mixed together. The mixture is screened, then moistened with a solution of polyvinylpyrrolidone in water, kneaded, wet-granulated and dried. The granules, the remaining corn starch and the magnesium stearate are screened and mixed together. The mixture is compressed to produce tablets of suitable shape and size.


B)	Tablets	per tablet

active substance	80	mg
lactose	55	mg
corn starch	190	mg
microcrystalline cellulose	35	mg
polyvinylpyrrolidone	15	mg
sodium-carboxymethyl starch	23	mg
magnesium stearate	2	mg
	400	mg

The finely ground active substance, some of the corn starch, lactose, microcrystalline cellulose and polyvinylpyrrolidone are mixed together, the mixture is screened and worked with the remaining corn starch and water to form a granulate which is dried and screened. The sodium carboxymethyl starch and the magnesium stearate are added and mixed in and the mixture is compressed to form tablets of a suitable size.


	C)	Ampoule solution

active substance	50	mg
sodium chloride	50	mg
water for inj.	5	ml

The active substance is dissolved in water at its own pH or optionally at pH 5.5 to 6.5 and sodium chloride is added to make it isotonic. The solution obtained is filtered free from pyrogens and the filtrate is transferred under aseptic conditions into ampoules which are then sterilised and sealed by fusion. The ampoules contain 5 mg, 25 mg and 50 mg of active substance.

Claims

1. A method of treatment of a disease or condition responsive to an inhibitor of PLK, said disease or condition being selected from the group consisting of: cancer, infection, inflammation and an autoimmune disease, said method comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound of the following formula (1):

wherein

W denotes N or C—R²,

X denotes —NR^1a, O or S,

Y denotes CH,

Z denotes —CF₃;

A is selected from one of the following formulas (i), (ii) and (iii):

Q₁denotes that (i), (ii) and (iii) are mono- or bicyclic aryl;

B¹, B², B³and B⁴each independently of one another denote C—R^gR^h, N—Rⁱ, O or S;

R¹and R^1aeach independently of one another denote hydrogen or methyl;

R²denotes one of hydrogen, halogen, —OR⁴, —C(═O)R⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —C═NRⁱ, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, pseudohalogen, and a mono- or polysubstituted group selected from the group consisting of C_1-6alkyl, C_2-6alkenyl, C_2-6alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of halogen, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen;

R^a, R^h, R^e, R^d, R^e, R^f, R^gand R^heach independently of one another denote a group selected from the group consisting of hydrogen, halogen, ═O, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —C═NRⁱ, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶, pseudohalogen, and an unsubstituted or mono- or polysubstituted group selected from the group consisting of C_1-6-alkyl, C_2-6-alkenyl, C_2-6-alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of halogen, R⁸, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen; wherein R^gand R^hare optionally located at the same or at adjacent C atoms and are attached in any combination to a common saturated or partially unsaturated 3-5-membered alkyl bridge which contains one to two heteroatoms;

Rⁱdenotes a group selected from the group consisting of hydrogen, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶, pseudohalogen and an unsubstituted or substituted mono- or polysubstituted group selected from the group consisting of C_1-6alkyl, C_2-6alkenyl, C_2-6alkynyl, C_3-6cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of halogen, R⁸, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen; wherein the Rⁱgroups located at adjacent N atoms are optionally joined together or Rⁱwith R^gor R^hlocated at adjacent C atoms are optionally attached in any combination to a common saturated or partially unsaturated 3-5-membered alkyl bridge which contains one to two heteroatoms;

R³is selected from the following formulas (iv)-(x):

R⁴, R⁵and R⁶each independently of one another denote hydrogen or an unsubstituted or mono- or polysubstituted group selected from the group consisting of C_1-5-alkyl, C_2-5alkenyl, C_2-5alkynyl, C_3-10cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of C_3-10-cycloalkyl, aryl, heterocyclyl, heteroaryl, halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;

L denotes a bond or an unsubstituted or mono- or polysubstituted group selected from the group consisting of C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;

Q₂and Q₃each independently of one another denote a bond or an unsubstituted or mono- or polysubstituted group selected from the group consisting of C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of halogen, —NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸C(═O)R⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR^B, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen;

R⁷denotes hydrogen or an unsubstituted or mono- or polysubstituted group selected from the group consisting of C_1-16-alkyl, C_2-16-alkenyl, C_2-16-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of halogen, NO₂, —OR⁸, —C(═O)R⁸, —C(═O)OR⁸, —C(═O)NR⁸R⁹, —NR⁸R⁹, —NR⁸COR⁹, —NR⁸C(═O)OR⁹, —NR⁸C(═O)NR⁹R¹⁰, —NR⁸C(═O)ONR⁹R¹⁰, —NR⁸SO₂R⁹, —N═CR⁸R⁹, —SR⁸, —SOR⁸, —SO₂R⁸, —SO₂NR⁸R⁹, —NR⁸SO₂NR⁹R¹⁰, —OSO₂NR⁸R⁹and pseudohalogen; and

R⁸, R⁹and R¹⁰each independently of one another denote hydrogen or a substituted or unsubstituted group selected from the group consisting of C_1-8-alkyl, C_2-8alkenyl, C_2-8-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the substituted group are identical or different and are selected from the group consisting of halogen, methyl, ethyl, amino, methylamino, dimethylamino, —OH and pseudohalogen;

or a pharmacologically acceptable acid addition salt thereof

in an excipient or carrier.

2. A method of prevention of a disease or condition responsive to an inhibitor of PLK, said disease or condition being selected from the group consisting of: infection, and inflammation; said method comprising administering a therapeutically effective amount of a pharmaceutical composition-comprising the compound the following formula (1):

wherein

W denotes N or C—R²,

X denotes —NR^1a, O or S,

Y denotes CH,

Z denotes —CF₃;

A is selected from one of the following formulas (i), (ii) and (iii):

Q₁denotes that (i), (ii) and (iii) are mono- or bicyclic aryl;

R¹and R^1aeach independently of one another denote hydrogen or methyl;

R²denotes one of hydrogen, halogen, —OR⁴, —C(═O)R⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —C═NRⁱ, —SR⁴, —SOW', —SO₂R⁴, —SO₂NR⁴R⁵, pseudohalogen, and a mono- or polysubstituted group selected from the group consisting of C_1-6alkyl, C_2-6alkenyl, C_2-6alkynyl, C_3-6-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the mono- or polysubstituted group are identical or different and are selected from the group consisting of halogen, —NO₂, —OR⁴, —C(═O)R⁴, —C(═O)OR⁴, —C(═O)NR⁴R⁵, —NR⁴R⁵, —NR⁴C(═O)R⁵, —NR⁴C(═O)OR⁵, —NR⁴C(═O)NR⁵R⁶, —NR⁴SO₂R⁵, —N═CR⁴R⁵, —SR⁴, —SOR⁴, —SO₂R⁴, —SO₂NR⁴R⁵, —NR⁴SO₂NR⁵R⁶, —OSO₂NR⁵R⁶and pseudohalogen;

R³is selected from the following formulas (iv)-(x):

R⁸, R⁹and R¹⁰each independently of one another denote hydrogen or a substituted or unsubstituted group selected from the group consisting of C_1-8-alkyl, C_2-8-alkenyl, C_2-8-alkynyl, C_3-10-cycloalkyl, aryl, heterocyclyl and heteroaryl, wherein the substituent(s) of the substituted group are identical or different and are selected from the group consisting of halogen, methyl, ethyl, amino, methylamino, dimethylamino, —OH and pseudohalogen;

or a pharmacologically acceptable acid addition salt thereof in an excipient or carrier.