HK1080840B - Arylcarbonylpiperazines and heteroarylcarbonylpiperazines and the use thereof for treating benign and malignant tumour diseases - Google Patents

Description

Arylcarbonylpiperazines and heteroarylcarbonylpiperazines and their use for the treatment of benign and malignant tumor diseases

In the coming years, there is a dramatic increase in the prediction of neoplastic diseases and tumor-related deaths worldwide. In 2001, about ten million people worldwide have cancer, more than six million people die from the disease. Tumor formation is a fundamental disease of the plant kingdom, the animal kingdom and the human higher organisms. The acquisition of a well-established multi-step carcinogenesis model postulates that as a result of the accumulation of a large number of individual cell mutations, the proliferation and differentiation behavior of cells is altered so that finally malignant stages with metastasis are reached via benign intermediate stages. Behind the name cancer or tumor, two hundred or more different clinical diseases are hidden. Neoplastic diseases can progress in either benign or malignant ways. The most important tumors are lung, breast, stomach, cervix, prostate, head and neck, large and small intestine, liver and blood system tumors. There are great differences with respect to course, prognosis and treatment behaviour. More than 90% of the actual cases involve solid tumors, which are refractory or untreatable, particularly in the advanced stages or in metastases. The three approaches to cancer control remain surgical resection, radiation, and chemotherapy. Despite major advances, it has not been possible to develop drugs that significantly prolong survival or, in turn, cure completely for widespread solid tumors. Therefore, it is very interesting to invent new drugs for cancer control.

The present invention relates to novel aryl-and heteroaryl-substituted piperazinylcarbonyl compounds and their homologues, their manufacture and use as medicaments, in particular for the treatment of benign and malignant tumours in humans and mammals.

Substituted and unsubstituted acridine-, quinoline-or pyridocarbonylpiperazines with anticancer properties are described, for example, in patent specifications WO 2002/008194, WO2002/008192 and WO 2002/008190 of the company Zentaris AG.

In patent specifications DE 1102747 and US 3843657, fluorene derivatives are described which have spasmolytic properties or have antibacterial and fungicidal properties. No effect on tumors is described nor suggested.

Xanthene derivatives are described in the literature as spasmolytic (US 2742472) and antiulcer (US 3284449). No effect on tumors is described nor suggested. Cinnoline derivatives of the above substance classes are mentioned in the literature as having different biological properties, for example as being anti-inflammatory (J.MeD.chem.1966, 9, 664) or as having CNS activity (A.Stanczak et al.Pharmazie 1997, 521, 91-97; US 3299070). No effect on tumors is described nor suggested.

Duro et al farnco, 1981, 36(6), 400-oza 411 describe isoquinoline derivatives and their use as local anesthetics. Furthermore, isoquinolines of the above structural type are used as antipyretics, antiarrhythmics and sedatives (DE 2811312, DE 2818423). Neither tumor activity is described nor suggested.

Isoxazoles and isothiazoles are described as effective antihypertensive agents in patent specifications US 4001237 and a. carenzi et al. arzneimittel forsch.1989, 39, 642. In addition, isoxazoles are described as fungicides (j.heindl et al eur.j.of med.chem.1975, 10, 591). Furthermore, isoxazoles have been demonstrated in the literature as analgesics (DE2065430), muscarinic receptor antagonists (h.g. striegel et al european j.of med.chem.1995, 30, 839), having antibacterial properties (a.pae et al biorg.med.chem.lett.1999, 18, 2679). Neither tumor activity is described nor suggested.

Pyrazoles are mentioned in the literature as compounds having anti-inflammatory and hypnotic properties (s. sugiura et al, j. med. chem.1977, 20, 80), anxiolytic agents (j.k. chakrabarti et al, j. med. chem.1989, 32, 2573), having antibacterial properties (g.palazzino et al, framaco ed. sci.1986, 41, 566), cannabinoid receptor antagonists (r.lau et al, j. med. chem.1999, 42, 769; r.pertwee et al, eur.j. pharmacol.1996, 296, 169), alpha-adrenoreceptor antagonists (g.ermanandi et al, farmoco.sci.1998, 53, 519), histamine H3 antagonists (WO 2003/004480), factor Xa inhibitors (WO 01/19798), sedatives and analgesics (EP 1006110), cholinesterase inhibitors (WO 98/39000 US 9720835) and CRF receptors (US 9720835). No effect on tumors is described nor suggested.

It has now surprisingly been found that novel compounds from the group consisting of aryl-and heteroaryl-substituted piperazinylcarbonylaromatic compounds are suitable for the preparation of medicaments, which are particularly suitable for the treatment of benign and malignant tumors. In accordance with this aspect, the present application claims novel compounds from the group consisting of aryl-and heteroaryl-substituted piperazinylcarbonyl compounds according to formula 1,

wherein the substituents have the following meanings:

r1: fluoren-9-one, isoxazole, cinnoline, isothiazole, isoquinoline, 9H-fluorene, 9H-xanthene and 1H-pyrazole,

wherein bonding can occur via any desired and possible ring member of the heteroaryl or aryl group, the aromatic and heteroaromatic compounds can be mono-or polysubstituted or unsubstituted;

R2：O、S；

r3: represents 1 or up to 16 substituents selected from: H. unsubstituted or substituted hydrocarbon radicals, halogens, COOH, CONH₂，

Wherein the substituents may be arranged ortho or geminal to the heterocyclic ring;

r4: unsubstituted or substituted aryl, unsubstituted or substituted heteroaryl, unsubstituted or substituted alkylaryl, unsubstituted or substituted alkylheteroaryl;

m、n：0-3。

"halogen" in the sense of the present invention comprises the halogen atoms fluorine, chlorine, bromine and iodine.

"metal" in the sense of the present invention comprises metal ions, such as sodium, potassium, lithium, magnesium, calcium, zinc and manganese ions.

"hydrocarbyl" in the sense of the present invention comprises acyclic saturated or unsaturated hydrocarbon radicals which may be branched or straight-chain and unsubstituted or mono-or polysubstituted having 1 to 20C atoms, i.e. C_1-20Alkyl (alkanyl), C_2-20Alkenyl and C_2-20-alkynyl. Here, alkenyl has at least one C-C double bond and alkynyl has at least one C-C triple bond. The hydrocarbon radical is advantageously chosen from the group comprising methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 2-hexyl, n-octyl, ethenyl, ethynyl, propenyl (-CH)₂CH＝CH₂；-CH＝CH-CH₃；-C(＝CH₂)-CH₃) Propynyl (-CH)₂-C≡CH；-C≡C-CH₃) Butenyl, butynyl, pentenyl, pentynyl, hexenyl, hexynyl, octenyl and octynyl.

For the purposes of the present invention, "cyclic hydrocarbon group" means cyclic hydrocarbons having from 3 to 12 carbon atoms, which may be saturated or unsaturated, unsubstituted or substituted. The cycloalkyl group may also be part of a bicyclic or polycyclic ring system.

"Heterocyclyl" represents a 3-, 4-, 5-, 6-, 7-or 8-membered cyclic organic group containing at least 1, optionally 2, 3, 4 or 5 heteroatoms, wherein the heteroatoms are the same or different, the cyclic group being saturated or unsaturated, but not aromatic, and may be unsubstituted or mono-or polysubstituted. The heterocyclic ring may also be part of a bicyclic or polycyclic ring system. Preferred heteroatoms are nitrogen, oxygen and sulfur. Preferably, the heterocyclyl group is selected from the group consisting of tetrahydrofuranyl, tetrahydropyranyl, pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl, wherein bonding to the compound of formula 1 may occur via any desired ring member of the heterocyclyl group.

"aryl" in the sense of the present invention denotes aromatic hydrocarbons, especially phenyl, naphthyl and anthracenyl. These groups may also be fused with other saturated, (partially) unsaturated or aromatic ring systems. Each aryl group may exist in unsubstituted or mono-or polysubstituted form, wherein the aryl substituents may be the same or different and may be located in any desired and possible position on the aryl group.

"heteroaryl" represents a 5-, 6-or 7-membered cyclic aromatic radical containing at least 1, optionally 2, 3, 4 or 5 heteroatoms, wherein the heteroatoms are identical or different, and the heterocycle may be unsubstituted or mono-or polysubstituted; where the heterocyclic ring is substituted, the heteroaryl substituents, which may be the same or different, are located at any desired and possible position on the heteroaryl ring. The heterocyclic ring may also be part of a bicyclic or polycyclic ring system. Preferred heteroatoms are nitrogen, oxygen and sulfur. Preferably, the heteroaryl group is selected from the group consisting of pyrrolyl, furanyl, thienyl, thiazolyl, triazolyl, tetrazolyl, oxazolyl, isothiazolyl, isoxazolyl, pyrazolyl, imidazolyl, pyridyl, pyrimidinyl, pyrazinyl, triazinyl, benzothiazolyl, indolyl, indolizinyl, quinolinyl, isoquinolinyl, cinnolinyl, quinazolinyl, quinoxalinyl, phthalazinyl, carbazolyl, phenazinyl, phenothiazinyl, purinyl, acridinyl, phenanthryl, wherein the bonding to the compound of formula 1 can occur via any desired and possible ring member of the heteroaryl group.

For the purposes of the present invention, "alkylcycloalkyl", "alkylheterocyclyl", "alkylaryl" or "alkylheteroaryl" means that alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl have the meanings as defined above, the cycloalkyl, heterocyclyl, aryl or heteroaryl group being bound via C_1-8-a hydrocarbon group is bonded to the compound of formula 1.

With "alkyl", "alkenyl"The term "substituted" in connection with "alkynyl" is understood in the sense of the present invention to mean that hydrogen is substituted by: F. cl, Br, I, CN, NH₂NH-alkyl, NH-cycloalkyl, NH-aryl, NH-heteroaryl, NH-alkylaryl, NH-alkylheteroaryl, NH-heterocyclyl, NH-alkyl-OH, N (alkyl)₂N (alkylaryl)₂N (alkyl heteroaryl)₂N (heterocyclic radical)₂N (hydrocarbyl-OH)₂、NO、NO₂SH, S-alkyl, S-cycloalkyl, S-aryl, S-heteroaryl, S-alkylaryl, S-alkylheteroaryl, S-heterocyclyl, S-alkyl-OH, S-alkyl-SH, S-S-alkyl, S-S-cycloalkyl, S-S-aryl, S-S-heteroaryl, S-S-alkylaryl, S-S-alkylheteroaryl, S-S-heterocyclyl, S-S-alkyl-OH, S-S-alkyl-SH, S-S-alkyl-C (O) -NH-heterocyclyl, OH, O-alkyl, O-cycloalkyl, O-alkylcycloalkyl, O-aryl, O-heteroaryl, S-alkylheteroaryl, S-alkyl-OH, S-S-alkyl-SH, S-S-alkyl-C (O) -NH-heterocyclyl, OH, O-alkyl, O-cycloalkylalkyl, O-aryl, S-cycloalkyl, S-, O-heteroaryl, O-alkylaryl, O-alkylheteroaryl, O-heterocyclyl, O-alkylheterocyclyl, O-alkyl-OH, O-alkyl-O-alkyl, O-SO₂-N (hydrocarbyl)₂、O-SO₂-OH、O-SO₂-O-hydrocarbyl, O-SO₂-O-cycloalkyl group, O-SO₂-O-heterocyclic hydrocarbon radical, O-SO₂-O-hydrocarbyl cycloalkyl group, O-SO₂-O-hydrocarbyl heterocyclic hydrocarbyl, O-SO₂-O-aryl, O-SO₂-O-heteroaryl, O-SO₂-O-alkylaryl, O-SO₂-O-alkylheteroaryl, O-SO₂Hydrocarbon radicals, O-SO₂Cycloalkyl, O-SO₂-heterocyclic hydrocarbon radicals, O-SO₂-hydrocarbyl cycloalkyl, O-SO₂-hydrocarbyl heterocyclic hydrocarbyl, O-SO₂Aryl, O-SO₂Heteroaryl, O-SO₂Alkyl aryl, O-SO₂(iii) hydrocarbyl heteroaryl, O-C (O) -hydrocarbyl, O-C (O) -cycloalkyl, O-C (O) -heterocycloalkyl, O-C (O) -hydrocarbyl cycloalkyl, O-C (O) -hydrocarbyl heterocycloalkyl, O-C (O) -aryl, O-C (O) -heteroaryl, O-C (O) -hydrocarbyl aryl, O-C (O) -hydrocarbyl heteroaryl, O-C (O) O-hydrocarbyl, O-C (O) O-cycloalkyl, O-C (O) O-heterocycloalkyl, O-C (O) O-hydrocarbyl cycloalkyl, O-C (O) O-hydrocarbyl heterocycloalkyl, O-C (O) O-aryl, O-C (O) O-heteroaryl, O-C (O) alkylaryl, O-C (O) O-alkylheteroaryl, O-C (O) NH-alkyl, O-C (O) NH-cycloalkyl, O-C (O) NH-heterocycloalkyl, O-C (O) NH-cycloalkylcycloalkyl, O-C (O) NH-alkylcycloalkylO-C (O) NH-hydrocycloalkyl, O-C (O) NH-aryl, O-C (O) NH-heteroaryl, O-C (O) NH-hydrocycloaryl, O-C (O) NH-hydrocycloheteroaryl, O-C (O) N (alkyl)₂O-C (O) N (cycloalkyl)₂O-C (O) N (heterocyclic hydrocarbon group)₂O-C (O) N (hydrocarbyl cycloalkyl)₂O-C (O) N (hydrocarbyl heterocycloalkyl)₂O-C (O) N (aryl)₂O-C (O) N (heteroaryl)₂O-C (O) N (alkylaryl)₂O-C (O) N (alkylheteroaryl)₂、O-P(O)(OH)₂O-P (O) (O-metal)₂O-P (O) (O-alkyl)₂O-P (O) (O-cycloalkyl)₂O-P (O) (O-aryl)₂O-P (O) (O-heteroaryl)₂O-P (O) (O-alkylaryl)₂O-P (O) (O-alkyl heteroaryl)₂O-P (O) (N-alkyl)₂(N-alkyl)₂O-P (O) (N-cycloalkyl)₂(N-cycloalkyl)₂O-P (O) (N-heterocyclic hydrocarbon group)₂(N-heterocyclic hydrocarbon group)₂O-P (O) (N-aryl)₂(N-aryl)₂O-P (O) (N-heteroaryl)₂(N-heteroaryl)₂O-P (O) (N-alkylaryl)₂(N-alkylaryl)₂O-P (O) (N-alkyl heteroaryl)₂(N-alkylheteroaryl)₂CHO, C (O) -alkyl, C (S) -alkyl, C (O) -aryl, C (S) -aryl, C (O) -alkylaryl, C (S) -alkylaryl, C (O) -heterocyclyl, C (O) -heteroaryl, C (O) -alkylheteroaryl, C (S) -heterocyclyl, CO (O) -alkylheteroaryl₂H、CO₂-hydrocarbon radical, CO₂-cycloalkyl, CO₂-heterocyclyl, CO₂Aryl, CO₂-heteroaryl, CO₂Alkyl aryl, C (O) -NH₂C (O) NH-alkyl, C (O) NH-aryl, C (O) NH-heterocyclyl, C (O) NH-alkylheterocyclyl, C (O) N (alkyl)₂C (O) N (alkylaryl)₂C (O) N (alkylheteroaryl)₂C (O) N (heterocyclyl)₂SO-hydrocarbyl, SO₂-hydrocarbyl, SO₂Aryl, SO₂Alkyl aryl, SO₂Heteroaryl, SO₂Alkyl heteroaryl, SO₂NH₂、SO₃H、CF₃CHO, CHS, alkyl, cycloalkyl, aryl, alkylaryl, heteroaryl, alkylheterocyclyl and/or heterocyclyl, wherein the substituents are polysubstitutedRadicals are understood to mean polysubstituted, e.g. di-or tri-substituted, on different or identical atoms, e.g. trisubstituted on the same C atom, as in CF₃、-CH₂CF₃Or in different positions, as in-CH (OH) -CH ═ CH-CHCl₂. Multiple substitutions may occur with the same or different substituents.

With regard to aryl, heterocyclyl, heteroaryl, alkylaryl and cycloalkyl, mono-or poly-substitution is understood in the sense of the present invention to mean that one or more hydrogen atoms of the ring system are mono-or poly-substituted, for example di-, tri-or tetra-substituted, by: F. cl, Br, I, CN, NH₂NH-alkyl, NH-aryl, NH-heteroaryl, NH-alkylaryl, NH-alkylheteroaryl, NH-heterocyclyl, NH-alkyl-OH, N (alkyl)₂NC (O) alkyl, N (alkylaryl)₂N (alkyl heteroaryl)₂N (heterocyclic radical)₂N (hydrocarbyl-OH)₂、NO、NO₂SH, S-alkyl, S-aryl, S-heteroaryl, S-alkylaryl, S-alkylheteroaryl, S-heterocyclyl, S-alkyl-OH, S-alkyl-SH, OH, O-alkyl, O-cycloalkyl, O-alkylcycloalkyl, O-aryl, O-heteroaryl, O-alkylaryl, O-alkylheteroaryl, O-heterocyclyl, O-alkylheterocyclyl, O-alkyl-OH, O-alkyl-O-alkyl, O-SO-aryl, O-alkylheteroaryl, S-alkyl-OH, S-heteroaryl, S-alkylheteroaryl, S-heterocyclyl, O-alkyl-O-alkyl, O-alkyl-O-heterocyclyl, O-SO-alkyl, O-aryl₂-N (hydrocarbyl)₂、O-SO₂-OH、O-SO₂-O-hydrocarbyl, O-SO₂-O-cycloalkyl group, O-SO₂-O-heterocyclic hydrocarbon radical, O-SO₂-O-hydrocarbyl cycloalkyl group, O-SO₂-O-hydrocarbyl heterocyclic hydrocarbyl, O-SO₂-O-aryl, O-SO₂-O-heteroaryl, O-SO₂-O-alkylaryl, O-SO₂-O-alkylheteroaryl, O-SO₂Hydrocarbon radicals, O-SO₂Cycloalkyl, O-SO₂-heterocyclic hydrocarbon radicals, O-SO₂-hydrocarbyl cycloalkyl, O-SO₂-hydrocarbyl heterocyclic hydrocarbyl, O-SO₂Aryl, O-SO₂Heteroaryl, O-SO₂Alkyl aryl, O-SO₂-alkylheteroaryl, O-C (O) -alkyl, O-C (O) -cycloalkyl, O-C (O) -heterocycloalkyl, O-C (O) -alkylcycloalkyl, O-C (O) -alkylheterocycloalkyl, O-C (O) -arylheteroaryl, O-C (O) -alkylaryl, O-C (O) -alkylheteroarylarylO-C (O) alkyl, O-C (O) cycloalkyl, O-C (O) O-cycloalkyl, O-C (O) heterocycloalkyl, O-C (O) hydrocycloalkyl, O-C (O) arylo, O-C (O) heteroaryl, O-C (O) alkylaryl, O-C (O) alkylheteroaryl, O-C (O) NH-alkyl, O-C (O) NH-cycloalkyl, O-C (O) NH-heterocycloalkyl, O-C (O) NH-hydrocycloalkyl, O-C (O) NH-aryl, O-C (O) NH-heteroarylalkyl, O-C (O) NH-alkylaryl, O-C (O) NH-alkylheteroaryl, O-C (O) N (alkyl)₂O-C (O) N (cycloalkyl)₂O-C (O) N (heterocyclic hydrocarbon group)₂O-C (O) N (hydrocarbyl cycloalkyl)₂O-C (O) N (hydrocarbyl heterocycloalkyl)₂O-C (O) N (aryl)₂O-C (O) N (heteroaryl)₂O-C (O) N (alkylaryl)₂O-C (O) N (alkylheteroaryl)₂、O-P(O)(OH)₂O-P (O) (O-metal)₂O-P (O) (O-alkyl)₂O-P (O) (O-cycloalkyl)₂O-P (O) (O-aryl)₂O-P (O) (O-heteroaryl)₂O-P (O) (O-alkylaryl)₂O-P (O) (O-alkyl heteroaryl)₂O-P (O) (N-alkyl)₂(N-alkyl)₂O-P (O) (N-cycloalkyl)₂(N-cycloalkyl)₂O-P (O) (N-heterocyclic hydrocarbon group)₂(N-heterocyclic hydrocarbon group)₂O-P (O) (N-aryl)₂(N-aryl)₂O-P (O) (N-heteroaryl)₂(N-heteroaryl)₂O-P (O) (N-alkylaryl)₂(N-alkylaryl)₂O-P (O) (N-alkyl heteroaryl)₂(N-alkylheteroaryl)₂CHO, C (O) -alkyl, C (S) -alkyl, C (O) -aryl, C (S) -aryl, C (O) -alkylaryl, C (S) -alkylaryl, C (O) -heterocyclyl, C (S) -heterocyclyl, CO (O) -heterocyclyl₂H、CO₂-hydrocarbon radical, CO₂Alkyl aryl, C (O) -NH₂C (O) NH-alkyl, C (O) NH-aryl, C (O) NH-heterocyclyl, C (O) N (alkyl)₂C (O) N (alkylaryl)₂C (O) N (alkylheteroaryl)₂C (O) N (heterocyclyl)₂SO-hydrocarbyl, SO₂-hydrocarbyl, SO₂Aryl, SO₂Alkyl aryl, SO₂Heteroaryl, SO₂Alkyl heteroaryl, SO₂NH₂、SO₃H、CF₃CHO, CHS, alkyl, cycloalkyl, aryl, alkylaryl, heteroaryl, alkylheterocyclyl and/or heterocyclyl, substituted on one or optionally different atoms (wherein one substituent may optionally be substituted per se). The polysubstitution in this case occurs with identical or different substituents.

If the compounds of the general formula 1 according to the invention have at least one asymmetric center, they can exist in the form of racemates, in the form of pure enantiomers and/or diastereomers or in the form of mixtures of these enantiomers and/or diastereomers. The mixture may exist as stereoisomers in any desired mixing ratio.

The compounds according to the invention may, if possible, exist in tautomeric form.

Thus, for example, the compounds of the general formula 1 according to the invention which have one or more chiral centers and are present in the form of racemates can be separated into their optical isomers, i.e. enantiomers or diastereomers, by methods known per se. The separation can be carried out by column separation on a chiral phase, or recrystallization from an optically active solvent, or use of an optically active acid or base, or derivatization with an optically active reagent, for example an optically active alcohol, followed by removal of the group.

The compounds of formula 1 according to the invention can be converted into salts with inorganic and organic acids if they have sufficiently basic groups, for example secondary or tertiary amines. Preferably, the compounds of general formula 1 according to the invention form pharmaceutically acceptable salts with the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, p-toluenesulfonic acid, carbonic acid, formic acid, acetic acid, sulfoacetic acid, trifluoroacetic acid, oxalic acid, malonic acid, maleic acid, succinic acid, tartaric acid, racemic acid, malic acid, embonic acid (embonic acid), mandelic acid, fumaric acid, lactic acid, citric acid, taurocholic acid, glutamic acid, or aspartic acid. In particular, hydrochloride, hydrobromide, sulfate, phosphate, methanesulfonate, toluenesulfonate, carbonate, bicarbonate, formate, acetate, sulfoacetate, trifluoromethanesulfonate, oxalate, malonate, maleate, succinate, tartrate, malate, pamoate, mandelate, fumarate, lactate, citrate and glutamate are formed. The stoichiometry of the salt of the compound according to the invention which is formed can in this case be an integer or a non-integer multiple of one.

The compounds of the general formula 1 according to the invention can be converted with inorganic and organic bases into their physiologically tolerable salts if they contain sufficiently acidic groups, for example carboxyl, sulfonic acid, phosphoric acid or phenolic groups. Possible inorganic bases are, for example, sodium hydroxide, potassium hydroxide, calcium hydroxide, and organic bases are, for example, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dibenzylethylenediamine and lysine. The stoichiometry of the salt of the compound according to the invention produced may be an integer or non-integer multiple of one herein.

Also preferred are solvates, in particular hydrates, of the compounds according to the invention, which can be obtained, for example, by crystallization from solvents or aqueous solutions. In this context, one, two, three or as many as possible solvent or water molecules can be combined with the compounds according to the invention to give solvates and hydrates.

Chemical species are known to form solids in various atomic states, which are referred to as polymorphs or variants. Different variants of polymorphic substances may differ widely in their physical properties. The compounds of general formula 1 according to the invention may exist in various polymorphic forms, some variants being possibly metastable herein.

According to a further embodiment, there are provided compounds of the general formula 1 according to the invention, in which R1, R2, R3, n and m have the abovementioned meanings and R4 represents phenyl, which is unsubstituted or is substituted by one to five identical or different (C3578)₁-C₆) Alkoxy substitution, in which the adjacent oxygen atom may also pass through (C1-C)₂) -an alkylene linkage.

According to a further embodiment, there is provided a compound according to formula 1, wherein R1, R2, R3, n and m have the above meanings, and R4 represents 3, 5-dimethoxyphenyl.

According to a further embodiment, there is provided a compound according to formula 1, wherein R1, R2, R3, n and m have the above meanings and R4 represents 3-methoxyphenyl.

The most preferred compounds according to formula 1 are found in the following list:

4- [4- (3, 5-Dimethoxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (1)

4- [4- (6-methylpyridin-2-yl) piperazine-1-carbonyl ] fluoren-9-one (2)

4- [4- (3-hydroxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (3)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (5-methyl-3-phenylisoxazol-4-yl) methanone (4)

Cinnolin-4-yl- [4- (3, 5-dimethylphenyl) piperazin-1-yl ] methanone (5)

Cinnolin-4-yl- [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (6)

(3, 5-Dimethylthioisothiazol-4-yl) - [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (7)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] isoquinolin-1-yl methanone (8)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-fluoren-1-yl) methanone (9)

(9H-fluoren-9-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (10)

(9H-fluoren-1-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (11)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (12)

[4- (3-methoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (13)

[4- (3-methoxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (14)

[4- (6-methylpyridin-2-yl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (15)

[4- (3-hydroxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (16)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - [1- (4-nitrophenyl) -5-trifluoromethyl-1H-pyrazol-4-yl ] methanone (17)

According to a further aspect of the invention, a process for the preparation of the compounds according to the invention is claimed, which process comprises reacting a carboxylic acid derivative of the general formula 2 with an amine of the general formula 3, in which formula 2R 1 and R2 have the meaning indicated above, and Y represents a leaving group, such as halogen, hydroxy, (C2)₁-C₆) Alkoxy, preferably methoxy and ethoxy, -O-tosyl, -O-mesyl, tetrazolyl or imidazolyl, in the general formula 3R 4, m and n have the abovementioned meanings,

r1: aryl, heteroaryl

Formula 2 formula 3

The reaction is optionally carried out using condensing agents and/or catalysts and diluents and auxiliaries to give the desired products of the formula 1.

Synthesis of Compounds according to the invention

The compounds of formula 1 can be obtained, for example, from the following scheme 1:

scheme 1

The method comprises the following steps:

the method 2 comprises the following steps:

the starting compounds 2 and 3 are commercially available or can be prepared by processes known per se. The starting materials 2 and 3 are valuable intermediate compounds which can be used for the preparation of the compounds of formula 1 according to the invention.

The solvents and auxiliaries optionally used and the reaction parameters used, such as reaction temperature and time, are known to the expert knowledge of the person skilled in the art.

The compounds of the general formula 1 according to the invention are suitable as active compounds in pharmaceuticals, in particular in antitumor agents, for the treatment of humans and mammals. The mammal can be a domestic animal such as a horse, cow, dog, cat, rabbit, sheep, and the like.

The medicinal action of the compounds according to the invention can be based, for example, on interactions with the tubulin system, which inhibit the polymerization of tubulin. In addition, further known and unknown mechanisms of action to control tumor cells are also conceivable.

According to a further aspect of the invention, there is provided a method of controlling tumours in humans and mammals, which method comprises administering to the human or mammal at least one compound of formula 1 according to the invention in an amount which is effective in the treatment of tumours. The effective dose of each compound according to the invention for administration therapy depends inter alia on the nature and stage of the neoplastic disease, the age and sex of the patient, the mode of administration and the duration of the treatment. The medicaments according to the invention can be administered as liquid, semi-solid and solid pharmaceutical dosage forms. The mode of administration is adapted to the respective dosage form: aerosols, powders and dusts (dusting powders), tablets, coated tablets, emulsions, foams, solutions, suspensions, gels, ointments, pastes, pills, crayons, capsules or suppositories.

Pharmaceutical dosage forms, in addition to at least one ingredient according to the invention, optionally contain excipients, such as, inter alia, solvents, dissolution promoters, solubilizers, emulsifiers, wetting agents, defoamers, gel formers, thickeners, film formers, binders, buffers, salt formers, drying agents, flow regulators, fillers, preservatives, antioxidants, colorants, mold release agents, lubricants, disintegrants, flavoring agents and flavors, depending on the dosage form employed. The choice of excipients and the amounts thereof depends on the chosen pharmaceutical dosage form and on the prescription, as is known to the person skilled in the art.

The medicament according to the invention can be administered in a suitable manner as follows: for the epidermal mode of skin, the dosage form is solution, suspension, emulsion, foam, ointment, paste or patch; buccal, lingual or sublingual via the buccal and lingual mucosa, in the form of tablets, lozenges, coated tablets, licks (linctus) or gargle; enterally via the gastric and intestinal mucosa in the form of tablets, coated tablets, capsules, solutions, suspensions or emulsions; rectally via the rectal mucosa, in the form of suppositories, rectal capsules or ointments; nasal administration via the nasal mucosa in the form of drops, ointments or sprays; pulmonary or inhalation via bronchial and alveolar epithelium, in the form of aerosol or inhalant; a conjunctival form via conjunctiva, in the form of eye drops, eye ointments, eye tablets, eye sheets, or eye lotions; intravaginal via the mucosa of the reproductive organs in the form of pessaries, ointments and rinses, intrauterine in the form of pessaries; intraurethral via ureter, the dosage form is a rinse, ointment, or medicated probe (medium); entering into artery of artery, and the dosage form is injection; the preparation form of the injection or the infusion is intravenous, and the preparation form of the injection or the infusion is paravenous; intradermal mode of entering the skin, and the dosage form is injection or implant; subcutaneous mode under skin, the dosage form is injection or implant; entering the intramuscular mode of the muscle, and the dosage form is injection or implant; entering into the peritoneum, and making into injection or infusion.

With regard to the actual therapeutic requirements, the pharmaceutical action of the compounds of the general formula 1 according to the invention can be delayed by suitable measures. This can be achieved chemically and/or pharmaceutically. Examples of achieving a prolonged action are the use of implants, liposomes, sustained release dosage forms, suspensions of nanoparticles and "prodrugs" of the compounds according to the invention, to form poorly soluble salts and complexes, or the use of crystalline suspensions.

The compounds of general formula 1 according to the invention can be used alone or in combination with other cytotoxic substances, such as cisplatin, carboplatin, doxorubicin, ifosfamide, cyclophosphamide, 5-FU, methotrexate, or in combination with immunomodulating agents or antibodies, in particular with signal transduction inhibitors, such as herceptin, glivec or iressa.

Particularly preferred medicaments in this context contain at least one compound from the following group of compounds according to the invention:

4- [4- (3, 5-Dimethoxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (1)

4- [4- (6-methylpyridin-2-yl) piperazine-1-carbonyl ] fluoren-9-one (2)

4- [4- (3-hydroxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (3)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (5-methyl-3-phenylisoxazol-4-yl) methanone (4)

Cinnolin-4-yl- [4- (3, 5-dimethylphenyl) piperazin-1-yl ] methanone (5)

Cinnolin-4-yl- [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (6)

(3, 5-Dimethylthioisothiazol-4-yl) - [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (7)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] isoquinolin-1-yl methanone (8)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-fluoren-1-yl) methanone (9)

(9H-fluoren-9-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (10)

(9H-fluoren-1-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (11)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (12)

[4- (3-methoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (13)

[4- (3-methoxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (14)

[4- (6-methylpyridin-2-yl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (15)

[4- (3-hydroxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (16)

[4- (3, 5-dimethoxyphenyl) piperazin-1-yl ] - [1- (4-nitrophenyl) -5-trifluoromethyl-1H-pyrazol-4-yl ] methanone (17),

either in the form of the free base or in the form of a physiologically tolerable acid salt.

Following the general procedure on which scheme 1 is based, the following compounds are synthesized, which are listed below with their respective chemical names attached. By means of melting points or by means of¹H-NMR spectroscopy and/or mass spectroscopy for the analytical identification of the compounds according to the invention.

The chemicals and solvents used are commercially available or synthesized from conventional suppliers (Acros, AvocaDo, AlDrich, Fluka, Lancaster, MaybriDge, Merck, Sigma, TCI, etc.).

The invention is illustrated in more detail, but without being restricted thereto, by means of the following examples.

Example 1 (reaction scheme 1 method 1)

4- [4- (3, 5-Dimethoxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (1)

A solution of 1g (4.12mmol) of 9-fluorenone-4-carbonyl chloride in 30ml of dimethylformamide was treated successively with 0.67g (6.59mmol) of N-methylmorpholine, 0.92g (4.12mmol) of 1- (3, 5-dimethoxyphenyl) piperazine and 2.36g (4.53mmol) of Py-BOP (1-benzotriazolyltriprolidiniophosphonium hexafluorophosphate). The mixture was stirred at room temperature for 12 hours, left overnight at room temperature, the dimethylformamide was distilled off in vacuo, and the residue was purified by means of a silica gel column (silica gel 60, MerckAG, DarmstaDt) eluting with a gradient of dichloromethane/methanol (95:5 v/v).

Yield: 1.4g (79.3% of theory)

M.p.：161℃

¹H-NMR(DMSO-d6)δ＝7.71-7.4(m，7H)，6.08(s，2H)，6.0(s，1H)，3.98-3.85(m，2H)，3.68(s，6H)，3.45-2.9(m，6H)ppm.

Example 2 (reaction scheme 1, method 1)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (12)

A solution of 3g (13.26mmol) of xanthene-9-carboxylic acid in 90ml of dimethylformamide was treated successively with 2.15g (21.2mmol) of N-methylmorpholine, 2.95g (13.26mmol) of 1- (3, 5-dimethoxyphenyl) piperazine and 7.59g (14.59mmol) of Py-BOP (1-benzotriazolyltriprolidiniophosphonium hexafluorophosphate). The mixture was stirred at room temperature for 12 hours, left overnight at room temperature, the dimethylformamide was distilled off in vacuo, and the residue was purified by means of a silica gel column (silica gel 60, MerckAG, DarmstaDt) eluting with a gradient of dichloromethane/methanol (95:5 v/v).

Yield: 2.88g (50.4% of theory)

M.p.：155℃

¹H-NMR(DMSO-d6)δ＝7.28(d，2H)，7.23(d，2H)，7.15(d，2H)，7.07(t，2H)，6.12(s，2H)，6.03(s，1H)，5.72(s，1H)，4.03(m，2H)，3.71(s，6H)，3.58(m，2H)，3.23-3.06(m，4H)ppm.

Example 3 (reaction scheme 1, method 2)

[4- (3-methoxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (14)

A solution of 3.03g (16.1mmol) of 1-phenyl-1H-pyrazole-5-carboxylic acid in 40ml of dimethylformamide is reacted with 13.56g (25.76mmol) of polymer-bonded N-benzoyl-N-cyclohexylcarbodiimide (1.66mmol/g) by warming to 60 ℃ and allowing the components to react with one another for 30 minutes. 2.48g (12.88mmol) of 1- (3-methoxyphenyl) piperazine was added thereto, and the mixture was reacted for another 4 hours. The resin was then cooled, the dimethylformamide was distilled off in vacuo and the residue was purified by means of a silica gel column (silica gel 60, Merck AG, DarmstaDt) eluting with a gradient of dichloromethane/methanol (95:5 v/v).

Yield: 0.75g (12.6% of theory)

¹H-NMR(DMSO-d6)δ＝7.82(s，1H)，7.54-7.46(m，4H)，7.4(t，1H)，7.11(t，1H)，6.73(d，1H)，6.46(m，1H)，6.41-6.38(m，2H)，3.72(m，5H)，3.33(m，2H)，3.10(m，2H)，2.82(m，2H)ppm.

In analogy to the synthetic route in scheme 1 (method 1 or 2), the following compounds of general formula 1 were synthesized:

formula 1

Example 4: 4- [4- (6-methylpyridin-2-yl) piperazine-1-carbonyl ] fluoren-9-one (2)

¹H-NMR(DMSO-d6)δ＝7.72(d，1H)，7.68(d，1H)，7.62(t，1H)，7.54(d，1H)，7.51-7.40(m，4-H)，6.6(d，1H)，6.55(d，1H)，3.95(m，1H)，3.87(m，1H)，3.7(m，2H)，3.52-3.25(m，4H)，2.28(s，3H)ppm.

Example 5: 4- [4- (3-hydroxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (3)

ESI-MS：385.1[M+H]

Example 6: [4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (5-methyl-3-phenylisoxazol-4-yl) methanone (4)

¹H-NMR(DMSO-d6)δ＝7.58(m，2H)，7.47(m，3H)，5.96(m，3H)，3.75-3.63(m，8H)，3.26(m，4H)，3.15(m，2H)，2.48(s，3H)ppm.

Example 7: cinnolin-4-yl- [4- (3, 5-dimethylphenyl) piperazin-1-yl ] methanone (5)

M.p.：114℃

¹H-NMR(DMSO-d6)δ＝9.45(s，1H)，8.58(d，1H)，8.04(m，1H)，7.96(m，2H)，6.58(s，2H)，6.48(s，1H)，3.95(m，2H)，3.34(m，2H)，3.28(m，2H)，3.05(m，2H)，2.21(s，6H)ppm.

Example 8: cinnolin-4-yl- [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (6)

¹H-NMR(DMSO-d6)δ＝9.43(s，1H)，8.58(d，1H)，8.05(m，1H)，7.95(m，2H)，7.45(t，1H)，6.63(d，1H)，6.54(d，1H)，3.90(m，2H)，3.72(m，2H)，3.48-3.2(m，4H)，2.3(s，3H)ppm.

Example 9: (3, 5-Dimethylthioisothiazol-4-yl) - [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (7)

¹H-NMR(DMSO-d6)δ＝7.45(t，1H)；6.65(d，1H)，6.57(d，1H)，3.8-3.3(m，8H)，2.66(s，3H)，2.58(s，3H)，2.32(s，3H)ppm.

Example 10: [4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] isoquinolin-1-yl methanone (8)

¹H-NMR(DMSO-d6)δ＝8.54(d，1H)，8.06(d，1H)，7.98(d，1H)，7.92(d，1H)，7.83(t，1H)，7.72(t，1H)，6.08(s，2H)，5.99(s，1H)，3.95(m，2H)，3.68(s，6H)，3.35(m，2H)，3.24(m，2H)，3.05(m，2H)ppm.

Example 11: [4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-fluoren-1-yl) methanone (9)

M.p.：148℃

¹H-NMR(DMSO-d6)δ＝7.98(d，2H)，7.94(d，2H)，7.58(d，1H)，7.48(t，1H)，7.4(t，1H)，7.35(t，1H)，7.28(d，1H)，6.10(s，2H)，5.99(s，1H)，3.88(s，2H)，3.82(m，2H)，3.67(s，6H)，3.41(m，2H)，3.28(m，2H)，3.08(m，2H)ppm.

Example 12: (9H-fluoren-9-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (10)

M.p.：162-163℃

¹H-NMR(DMSO-d6)δ＝7.86(d，2H)，7.37(d，2H)，7.32(t，2H)，7.22(t，2H)，7.03(t，1H)，6.46(m，1H)，6.38(s，1H)，6.30(d，1H)，5.32(s，1H)，3.95-3.42(m，7H)，3.25-3.0(m，4H)ppm.

Example 13: (9H-fluoren-1-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (11)

M.p.：124℃

¹H-NMR(DMSO-d6)δ＝7.99(d，1H)，7.96(d，1H)，7.61(d，1H9，7.48(t，1H)，7.42(t，1H)，7.35(t，1H)，7.29(d，1H)，7.12(t，1H)，6.54(m，1H)，6.48(s，1H)，6.39(m，1H)，3.89(s，2H)，3.83(m，2H)，3.71(s，3H)，3.41(m，2H)，3.27(m，2H)，3.08(m，2H)ppm.

Example 14: [4- (3-methoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (13)

M.p.：110℃

¹H-NMR(DMSO-d6)δ＝7.30(t，2H)，7.22(t，2H)，7.15-7.05(m，5H)，6.56(d，1H)，6.48(d，1H)，6.4(d，1H)，5.74(s，1H)，4.05(m，2H)，3.74(s，3H)，3.58(m，2H)，3.2-3.06(m，4H)ppm.

Example 15: [4- (6-methylpyridin-2-yl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (15)

¹H-NMR(DMSO-d6)δ＝7.83(s，1H)，7.55-7.37(m，6H)，6.74(d，1H)，6.57(d，1H)，6.53(d，1H)，3.68(m，2H)，3.48(m，2H)，3.32(m，2H)，3.18(m，2H)，2.32(s，3H)ppm.

Example 16: [4- (3-hydroxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (16)

¹H-NMR(DMSO-d6)δ＝9.2(s，1H)，7.82(d，1H)，7.53-7.46(m，4H)，7.4(t，1H)，6.98(t，1H)，6.73(d，1H)，6.33(m，1H)，6.23(m，2H)，3.68(m，2H)，3.35(m，2H)，3.05(m，2H)，2.75(m，2H)ppm.

Example 17: [4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - [1- (4-nitrophenyl) -5-trifluoromethyl-1H-pyrazol-4-yl ] methanone (17)

¹H-NMR(DMSO-d6)δ＝8.45(d，2H)，8.18(s，1H)，7.88(d，2H)，6.1(s，2H)，6.0(s，1H)，3.77(m，2H)，3.69(s，6H)，3.53(m，2H)，3.2(m，2H)，3.12(m，2H)ppm.

Most preferred compounds of the invention are bases of the substance of formula 1 or their pharmaceutically acceptable salts selected from the group consisting of:

4- [4- (3, 5-Dimethoxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (1)

4- [4- (6-methylpyridin-2-yl) piperazine-1-carbonyl ] fluoren-9-one (2)

4- [4- (3-hydroxyphenyl) piperazine-1-carbonyl ] fluoren-9-one (3)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (5-methyl-3-phenylisoxazol-4-yl) methanone (4)

Cinnolin-4-yl- [4- (3, 5-dimethylphenyl) piperazin-1-yl ] methanone (5)

Cinnolin-4-yl- [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (6)

(3, 5-Dimethylthioisothiazol-4-yl) - [4- (6-methylpyridin-2-yl) piperazin-1-yl ] methanone (7)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] isoquinolin-1-yl methanone (8)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-fluoren-1-yl) methanone (9)

(9H-fluoren-9-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (10)

(9H-fluoren-1-yl) - [4- (3-methoxyphenyl) piperazin-1-yl ] methanone (11)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (12)

[4- (3-methoxyphenyl) piperazin-1-yl ] - (9H-xanthen-9-yl) methanone (13)

[4- (3-methoxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (14)

[4- (6-methylpyridin-2-yl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (15)

[4- (3-hydroxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (16)

[4- (3, 5-Dimethoxyphenyl) piperazin-1-yl ] - [1- (4-nitrophenyl) -5-trifluoromethyl-1H-pyrazol-4-yl ] methanone (17)

Biological Effect of the Compounds according to the invention

In vitro experiments on selected tumor models showed the following pharmacological activities.

Example 18: anti-proliferative effect on various tumor cell lines

The antiproliferative activity of the substances according to the invention was investigated in proliferation assays on established tumor cell lines. The assay used to determine the activity of the cell dehydrogenase makes it possible to determine the viability of the cells and indirectly the number of cells. The cell lines used were the human cervical cancer cell line KB/HeLa (ATCC CCL17), the ovarian adenocarcinoma cell line SKOV-3(ATCC HTB77), the human glioblastoma cell line SF-268(NCI 503138) and the lung cancer cell line NCI-H460(NCI 503473). In addition, for the investigation of the cell cycle-specific action of the substance, the RKOp27 cell system (M.Schmidt et al. oncogene 19 (20): 2423-9, 2000) was also used. RKO is a human colon cancer cell line in which cell cycle inhibitor p27 is induced by an ecdysone expression system^kip1Expressed, and specifically caused cell cycle arrest at the G2 phase. The non-specifically acting substance inhibits proliferation regardless of whether RKO cells are arrested in stage G1 or G2. However, cell cycle specific substances, such as tubulin inhibitors, are cytotoxic only when the cell cycle is not stopped and passed. In Table 1, the cytotoxic and/or growth inhibitory activity of the compounds is shown with/without p27^kip1Expression of (2). The compounds tested showed p27^kip1Has no cytotoxic activity. The results show that the compounds according to the invention have a very potent inhibitory effect on the proliferation of selected tumor cell lines.

Table 1: proliferation inhibition of selected compounds on human tumor cell lines in XTT cytotoxicity assays

n.c.: is not carried out

Example 19: inhibition of tubulin polymerization

Selected compounds were tested for inhibition of bovine tubulin polymerization in vitro assays. In this experiment, tubulin purified by polymerization and depolymerization cycles was used, GTP was added and polymerization was initiated by heating. Watch (A)2, inhibition of polymerization of tubulin containing 30% of the protein of interest (MAP) and tubulin without MAP, EC is indicated₅₀The value is obtained. The results show that the substances according to the invention have a good to very good inhibitory effect on tubulin polymerization.

Table 2: inhibition of tubulin polymerization. Average of two independent experiments

n.c.: is not carried out

Description of the methods used

XTT assay for cellular dehydrogenase activity

Under standard conditions, 5% CO at 37 deg.C₂And fumigated incubators at 95% atmospheric humidity, the tumor cell lines KB/HeLa, SKOV-3, SF-268 and NCI-H460, which grow adherently, were cultured. On day 1 of the experiment, cells were detached using trypsin/EDTA and pelleted by centrifugation. Subsequently, the cell pellet was resuspended in the respective medium in the corresponding cell number and reacted in a 96-well microtiter plate. The plates were then incubated overnight in a fumigator. Stock solutions of the test substances at 1mg/ml in DMSO were prepared and diluted with the medium to the appropriate concentration on day 2 of the experiment. The medium was then added to the cells and incubated in a fumigated incubator for 45 hours. As a control, cells not treated with the test substance were used. For the XTT assay, 1mg/ml XTT (3' - [1- (phenylaminocarbonyl) -3, 4-tetrazolium)]Bis (4-methoxy-6-nitro) benzenesulfonic acid) was dissolved in RPMI-1640 medium without phenol red. In addition, a Phosphate Buffered Saline (PBS) solution of PMS (N-methyldibenzopyrazine methylsulfate) at 0.383mg/ml was prepared. On experiment day 4, 75 μ l/well of the XTT-PMS mixture was pipetted onto the cell plate, during which the plate had been incubated with the test substance for 45 hours. For this purpose, shortly before use, the XTT solution is mixed with the PMS solution at 50:1(vol: vol). Then the cell plate is cultured in a fumigation incubatorFor another 3 hours, the Optical Density (OD) was measured in a photometer_490nm). By means of the determined OD_490nmPercent inhibition relative to control was calculated and concentration-effect curves were plotted in a semi-logarithmic manner. EC was calculated by means of regression analysis of the concentration-action curve using the Graphpad Prism program₅₀。

Cell cycle analysis by means of the RKOp27 model

The assay is performed in 96 well plates. By induction p27^kip1Expression, cells completely stopped growing but did not die. By comparing the activity on induced and non-induced cells, conclusions can be drawn about the mechanism of action (cell cycle specificity) of the therapeutic agent. Non-induced cells were seeded at approximately three times higher cell number, since differentiation no longer occurred during the assay, whereas non-induced cells did not (20000 induced cells/well, 6250 non-induced cells/well). Controls were untreated cells (+/-induction). Induction was performed with 3. mu.M muristerone A. On day 1, cells were exposed to +/-muristerone A and incubated at 37 ℃ for 24 hours. On day 2, test substances (control DMSO) were added, incubation at 37 ℃ continued for an additional 45 hours, and then standard XTT assays were performed.

Tubulin polymerization assay

The assay was carried out based on the method of Bollag et al. Lyophilized bovine tubulin (cytoskeleton, ML113 tubulin with 30% MAP, TL238 tubulin without MAP) was solubilized at a concentration of 2mg/ML (ML 113 in 80mM PIPES, 0.5mM EGTA, 2mM MgCl)₂pH6.9, 1mM GTP) or 5mg/ml (TL 238 dissolved in 80mM PIPES, 1mM EGTA, 0.5mM MgCl₂20% (v: v) glycerol, pH6.9, 1mM GTP). The test substance was diluted in 10% DMSO (v: v), and 5. mu.l of the dilution was transferred to a 96-well microtiter plate (Nunc, half-area plate). After addition of 45. mu.l of tubulin solution, polymerization at 340nm was determined by means of a kinetic program in a Spectramax 190 microtitre plate reader (Molecular Devices) at 30 sec intervals over 20 min. The resulting area under the curve values were used to calculate the values for the untreated pairsConcentration-effect curves were plotted in a semi-logarithmic manner against inhibition. EC was calculated by means of regression analysis of the concentration-action curve using the Graphpad Prism program₅₀。

Examples of pharmaceutical administration forms

Example I

Tablets containing 50mg of active compound

Consists of the following components:

(1) active Compound 50.0mg

(2) Lactose 98.0mg

(3) Corn starch 50.0mg

(4) Polyvinylpyrrolidone 15.0mg

(5) Magnesium stearate 2.0mg

In total: 215.0mg

Preparation:

mixing (1), (2) and (3), and granulating with the aqueous solution of (4). Mixing the dried granules with the mixture of (5). Tablets were compressed from this mixture.

Example II

Capsule containing 50mg of active compound

Consists of the following components:

(1) active Compound 50.0mg

(2) Dried corn starch 58.0mg

(3) Powdered lactose 50.0mg

(4) Magnesium stearate 2.0mg

In total: 160.0mg

Preparation:

trituration of (1) with (3). This triturated product was added to the mixture of (2) and (4) and mixed well. This powder mixture was filled into size 3 hard gelatin capsules on a capsule filling machine.

Claims (12)

1. Heteroaryl-substituted piperazinylcarbonyl compounds of the general formula (1),

wherein the substituents have the following meanings:

r1: 1H-pyrazole, which is a compound of the formula,

wherein the bonding may occur via any desired and possible ring member of said heteroaryl group, and said heteroaryl group may be mono-or poly-substituted or unsubstituted;

R2：O、S；

r3: represents 1 or up to 8 substituents selected from: h or a halogen, or a salt thereof,

wherein the substituents may be arranged ortho or geminal to the heterocyclic ring;

r4: unsubstituted or substituted aryl, said aryl being selected from phenyl, naphthyl and anthracenyl; unsubstituted or substituted heteroaryl selected from pyridine;

m、n：1；

wherein:

"substituted" in connection with aryl, heteroaryl means that one or more hydrogen atoms of the ring system are replaced by: F. cl, Br, I, NH₂、NO₂S-hydrocarbyl, OH, O-hydrocarbyl, O-P (O) (OH)₂、CF₃Alkyl, aryl, heteroaryl, wherein the substituents are the same or different and may be present at any desired and possible position in the aryl, heteroaryl group, and wherein the polysubstituted groups may be present in the same or different substituents, on different or the same atoms;

"halogen" is the halogen atoms fluorine, chlorine, bromine and iodine;

"aryl" means an aromatic hydrocarbon, unsubstituted or mono-or polysubstituted, selected from phenyl, naphthyl and anthracenyl;

"heteroaryl" represents a 5-, 6-or 7-membered cyclic aromatic radical which is unsubstituted or mono-or polysubstituted, identical or different, and which contains at least 1 heteroatom nitrogen,

the "hydrocarbon group" is a branched or straight chain acyclic saturated hydrocarbon group having 1 to 20C atoms.

2. Compounds of the general formula (1) as claimed in claim 1, in which the heteroaryl group contains optionally 2 or 3 heteroatom nitrogens, whose bonding to the compounds of the general formula (1) can take place via the heteroaryl group, as desired, with possible ring members, where the heterocyclic ring can also be part of a bicyclic or polycyclic ring system.

3. The heteroarylcarbonylpiperazine compound of formula (1) as claimed in claim 1 or 2, wherein the alkyl group may be methyl, ethyl, n-propyl, 2-propyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 2-hexyl, n-octyl.

4. Compounds of the general formula (1) as claimed in claim 1 or 2, wherein R1, R2, R3, n and m have the meanings indicated in claim 1, and R4 represents phenyl, which is unsubstituted or substituted by one to five identical or different (C3534)₁-C₆) Alkoxy substitution, in which adjacent oxygen atoms may also pass through (C)₁-C₂) -an alkylene linkage.

5. Compounds of the general formula (1) as claimed in claim 1 or 2, wherein R1, R2, R3, n and m have the meanings indicated in claim 1 and R4 represents 3, 5-dimethoxyphenyl.

6. Compounds of the general formula (1) as claimed in claim 1 or 2, wherein R1, R2, R3, n and m have the meanings indicated in claim 1 and R4 represents 3-methoxyphenyl.

7. Physiologically tolerable salts of the compounds of the formula (1) as claimed in claims 1 to 6 consist of basic compounds neutralized with inorganic and organic acids or acidic compounds neutralized with inorganic and organic bases.

8. A compound of general formula (1) according to claim 1 or 2, which is one of the following compounds:

[4- (3-methoxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (14)

[4- (6-methylpyridin-2-yl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (15)

[4- (3-hydroxyphenyl) piperazin-1-yl ] - (2-phenyl-2H-pyrazol-3-yl) methanone (16)

[4- (3, 5-dimethoxyphenyl) piperazin-1-yl ] - [1- (4-nitrophenyl) -5-trifluoromethyl-1H-pyrazol-4-yl ] methanone (17).

9. A process for the preparation of a heteroarylcarbonylpiperazine compound as claimed in claim 1 or 2, which comprises reacting a carboxylic acid of the general formula 2 with an amine of the general formula 3, in which formula 2R 1 and R2 have the meanings given above in claim 1 and Y represents a leaving group selected from halogen, hydroxy, (C1-C6) -alkoxy, -O-tosyl, -O-mesyl, tetrazolyl or imidazolyl, in which formula 3R 4, m and n have the meanings given in claim 1,

r1: aryl, heteroaryl

Formula 2 formula 3

The reaction optionally uses condensing agents and/or catalysts, as well as diluents and auxiliaries, to give the desired product.

10. Use of the heteroarylcarbonylpiperazine compounds of general formula (1) as claimed in one of claims 1 to 6 as therapeutically active compounds for the preparation of medicaments for the treatment of tumors in humans and mammals.

11. Medicament for the treatment of tumors in humans and mammals, comprising a compound of general formula (1) as claimed in one of claims 1 to 6, together with customary pharmaceutically tolerable excipients, additives and carriers.

12. A process for the preparation of a medicament as claimed in claim 11, which process comprises processing one or more heteroarylcarbonylpiperazine compounds of the general formula (1) as claimed in any of claims 1 to 6 with customary pharmaceutical carriers and/or diluents or other excipients to give a pharmaceutical preparation or bringing them into a therapeutically administrable dosage form.