DE10253720A1 - New 10-phenylimino-9(10H)-anthracenone derivatives are tubulin polymerization inhibitors, useful for the treatment of hyperproliferative diseases - Google Patents

Abstract

10-Phenylimino-9(10H)-anthracenones (I) and their 1,8-dichloro derivatives (II) are new. 10-Phenylimino-9(10H)-anthracenones of formula (I) and their 1,8-dichloro derivatives (II) are new. R1-R4 = H, 1-6C alkoxy, 1-6C alkyl, aryloxy, COOH, SO3H, SO3M, ester, thioester, CN, acyl, aryloxy, benzyloxy, NO2, OH, NH2, SH, halo, OP(O)(OM)2, NP(O)(OM)2 or NP(O)(OR)2; M = a cation; and R = 1-8C alkyl, benzyl or aryl.

Description

The present invention relates to novel inhibitors of tubulin polymerization. It is about around 10-phenylimino-9 (10H) -anthracenones and their derivatives as anti-tumor effective connections.

Cancer poses one of the biggest health threats of humanity, and therefore considerable efforts are being made the development of new chemotherapy drugs for a more potent and specific one anti-cancer therapy undertaken. The most striking and medically significant A characteristic of cancer cells is their uncontrolled proliferation. The cellular Proliferation is a result of cell division or mitosis.

Microtubules are extremely important for a wide variety of cell functions, including mitosis, cell movement, customs form, and also for the transport of organelles within the cell. The microtubules are a separate class of cytoskeletal components. The importance of microtubules for cellular life and especially for cell division and the associated induction and progression of apoptosis make them one of the most attractive therapeutic targets for the development of new compounds with importance for anti-cancer chemotherapy. Tubulin together with its isoforms forms the main component of the microtubules and in turn exists as a heterodimer of the two globular polypeptide subunits α- and β-tubulin. Both peptides have a molecular mass between 50-55. Under physiological conditions the tubulin polymerizes to microtubules. As the αβ dimer is incorporated into the microtubule, the heterodimer binds two molecules of guanosine triphosphate (GTP) and hydrolyzes one of them to form GDP and inorganic phosphate. Electron microscopic examinations and X-ray structure analyzes confirmed the structure of the microtubule from 13 parallel, staggered protofilaments that enclose a cavity. Microtubules are constantly being built up and broken down, they show dynamic instability. It has been shown for numerous naturally occurring antimitotic compounds that they exert their effect by binding to the tubulin. The so-called MAPs ("microtubule-associated proteins") or other proteins involved in the mitotic process play a rather subordinate role here. There are three main classes of antimitotic compounds. There are microtubule-stabilizing compounds, substances that are associated with Vinca -Alkaloid binding site bind as well as substances attacking the colchicine binding site.The former act by binding and thus blocking the depolymerization, the latter two classes of compound, however, exert their effect primarily by inhibiting tubulin polymerization. Some clinically promising substances with documented cytotoxic and anti-tumor effects are due to an interaction with the process of microtubule assembly-disassembly or microtubule dynamics, which ultimately leads to a blockage of the cell cycle in the mitosis phase and to the induction of apoptosis, among the most important anti-micro The vinca alkaloids, such as vinblastine and vineristin, as well as the newer taxanes such as paclitaxel (taxol) and docetaxel (taxotere), which have been successfully introduced into clinical oncology, are tubule-active substances for cancer therapy. Colchicine is another important antimitotic substance. Although colchicine has only very limited medical applicability due to the very small therapeutic window, the substance played a fundamental role in elucidating the properties and functions of tubulin and microtubules. Furthermore, the natural substances rhizoxin ¹ , combretastatin A-4 and A-2 ^2,3 , curacin A, podophyllotoxin ⁴ , epothilones A and B ⁵ and dolastatin ⁶ as well as some synthetic compounds such as phenstatin, 2-styrylquinazolin-4 ( 3H) -one ⁷ and highly oxygenated derivatives of cis- and trans-stilbenes ⁸ exert their cytotoxic properties through a binding interaction with the tubulin. In many cases, the specific interactions with the binding site are not fully known and are largely unknown for a large number of tubulin-binding substances.

The present invention relates to new ones Inhibitors of tubulin polymerization with phenylimino-9 (10H) -anthracenone structure as well as their analogues. It was surprisingly found that certain 10-phenylimino-9 (10H) anthracenones were potent Inhibitors of tubulin polymerization are and therefore for treatment of diseases that are associated with increased cellular proliferation accompanied. The tubulin-binding according to the invention Substances can thus, for example, as effective anti-cancer compounds act.

Short description of the pictures

The following illustrations are part of it this patent and serve your better understanding as well the specification of the current Invention.

1 shows a representative reaction scheme for the synthesis of the 10-phenylimino-9 (10H) -anthracenones according to the invention and the 1,8-dichloro analogues.

2 shows the effect of compound A143 on the growth of human chronic myelogenic K562 cells. The cells were treated with different concentrations of the compound over a period of 48 h and the cell number was determined by counting using a hemocytometer (Neubauer chamber). The 100% value corresponds to a cell number of 1.20 × 10 ⁶ K562 cells per ml. The values represent the mean ± SD (standard deviation) from six samples; Bar, SD.

3 shows the chemosensitivity profile of compounds A145 and A146 against selected tumor cell lines propidium iodide assay). Bars pointing left: more sensitive cell lines, bars pointing right: more resistant cell lines.

4 shows the assembly of the tubulin in PIPES buffer at pH = 6.8 and 37 ° C. The tubulin concentration is 1.1 × 10 ^-5 M.

Detailed Description of the invention

worin die erfindungsgemäßen Verbindungen einen oder mehrere Substituenten (R1–R5) enthalten. Bevorzugte Verbindungen gemäß der Erfindung sind die in Tabelle I dargestellten sowie die in den Beispielen exemplarisch aufgeführten Verbindungen. Eine besonders bevorzugte Verbindung der allgemeinen Formel I ist diejenige mit R2 = OH and R3 = OCH₃. Eine besonders bevorzugte Verbindung der allgemeinen Formel II ist diejenige mit R2 = OH and R3 = OCH₃.The invention relates to 10-phenylimino-9 (10H) -anthracenones and 1,8-dichloro-10-phenylimino-9 (10H) -anthracenones of the general formulas I and II,

wherein the compounds of the invention contain one or more substituents (R1-R5). Preferred compounds according to the invention are the compounds shown in Table I and the compounds listed by way of example in the examples. A particularly preferred compound of the general formula I is that with R2 = OH and R3 = OCH ₃ . A particularly preferred compound of the general formula II is that with R2 = OH and R3 = OCH ₃ .

The following are representative of the compounds according to the invention:
In accordance with the present invention, in the phenylimino-9 (10H) anthracenone structure, the terminal phenyl residue can be replaced (modified) with a structure having a terminal heterocyclic group such as thiophene, pyrrole or imidazole.

10- (3 ', 5'-Dimethoxyphenylimino) -9 (10H) -anthracenone; 10- (3 ', 4'-Dimethoxyphenylimino) -9 (10H) -anthracenone; 10- (4'-methoxyphenylimino) -9 (10H) -anthracenone; 10- (3'-hydroxy-4'-methoxy-phenylimino) -9 (10H) -anthracenone; 10- (3 ', 4', 5'-Trimethoxyphenylimino) -9 (10H) -anthracenone; 10- (2 ', 4'-Dimethoxyphenylimino) -9 (10H) -anthracenone; 1,8-dichloro-10- (3-hydroxy-4-methoxy-phenylimino) -9 (10H) -anthracenone; 1,8-dichloro-10- (4'-methoxyphenylimino) -9 (10H) -anthracenone;

antineoplastic effect

The compounds of the invention have one anti-tubulin effect by inhibiting tubulin assembly (polymerization). The tubulin binding agents according to the invention Compounds are therefore useful as new anti-cancer compounds.

Salts The invention also includes the pharmaceutically acceptable acid and base addition salts of the compounds for pharmaceutical use. The pharmaceutically acceptable salts can be base addition salts. These include salts of the compounds with metals and amines, such as alkali or alkaline earth metals or organic amines. Examples of useful metal cations would be sodium, potassium, magnesium, calcium and the like. Also included are heavy metal salts with silver cations, Zinc, cobalt, and cerium. Examples of useful amines are N, N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine, and procaine. Pharmaceutically acceptable acid addition salts of the compounds become those formed with organic and inorganic acids. Examples of suitable acids for salt formation are hydrochloric acid, sulfuric acid, phosphoric acid, acetic acid, citric acid, oxalic acid, malonic acid, salicylic acid, malic acid, gluconic acid, fumaric acid, succinic acid, ascorbic acid, maleic acid, methanesulfonic acid, and the like. The salts can be obtained by treating the free basic form with a sufficient amount of the desired acid to form a mono salt, disalt, etc. in a conventional manner. The free basic form can be regenerated by treating a salt with a base. For example, dilute solutions of the aqueous base can be used. Dilute aqueous sodium hydroxide, potassium carbonate, ammonia and sodium hydrogen carbonate are useful for this purpose. The free basic forms differ from the salts in some physical properties such as, for example, their solubility in polar solvents, but the salts behave in accordance with the basic forms in the sense of the invention.

depolymerization by tubulin

The compounds of the invention bind Tubulin. By binding the tubulin-binding substances according to the invention there is an inhibition of tubulin assembly (polymerization). Suitable assays for determining the tubulin activity of the compounds according to the invention can be found in the given examples.

treatment proliferative diseases

The compounds of the invention have proven to be potent inhibitors of tubulin polymerization. she are therefore also suitable for inhibiting cell division and proliferation of non-cancer cells. Such diseases include EBV-induced lymphoproliferative diseases and lymphomas, proliferative secondary effects associated with diabetes, including vascular proliferation and retinopathy and psoriasis; further benign tumors, including angiomas, Fibromas, fibroids, osteoporosis, mastocytosis and myeloproliferative Diseases.

Anti-tumor effect

The compounds of the invention are useful for the purpose of tumor treatment, for example to inhibit the To achieve tubulin assembly of the tumor cell tubulin, an inhibition achieve tumor cell growth, kill tumor cells, apoptosis to induce and / or extend the life of a patient.

The carcinogenic, tubulin-binding compounds of the invention are useful for use in mammals. mammal in the sense of this invention denotes each class of higher vertebrates, who feed their young with milk produced in the mammary glands, including the Humans, rabbits and monkeys.

The inhibitors of tubulin polymerization according to the invention are suitable for the treatment of any disease in which the polymerization of tubulin is important. The compounds of the invention are also suitable for the treatment of diseases caused by parasitic protozoa, such as sporozoa (e.g. Plasmodium, Toxoplasma), amoebas (e.g. Acanthamoeba, Entamoeba), Flagellates (e.g. Trypanosoma, Leishmania), ciliates (e.g. Balantidium) be evoked and for which is important for the movement of tubulin. The inhibitors according to the invention tubulin polymerization are particularly useful for treating Chagas disease or diseases caused by worms.

Methods of Administration of the compounds according to the invention

The compounds of the invention can either as individual therapeutic agents or as a mixture with others therapeutic agents are administered. Generally will they are administered in the form of pharmaceutical agents, that is to say as mixtures of the active ingredients with suitable carriers or diluents. The compounds or means can oral, parenteral (e.g. intravenous, intraperitoneally), by inhalation or topically (including dermally, transdermal, buccal, and sublingual).

The type of pharmaceutical agent and the pharmaceutical carrier or diluent depends on the desired mode of administration. Oral agents can, for example, be in the form of tablets or capsules, also in retarded form, and can contain conventional excipients such as binders (e.g. syrup, acacia, gelatin, sorbitol, carrier type or polyvinylpyrrolidone), fillers (e.g. lactose, sucrose, corn starch, calcium phosphate, sorbitol or Glycine), lubricants (e.g. magnesium stearate, talc, polyethylene glycol or Silicon dioxide), disintegrating agents (e.g. starch) or wetting agents (e.g. sodium lauryl sulfate). Oral liquid preparations can be in the form of aqueous or oily suspensions, solutions, emulsions, syrups, elixirs or sprays etc. or can be dry powder for reconstitution with water or other suitable vehicle. Such liquid preparations can contain customary additives, for example suspending agents, flavoring agents, diluents or emulsifiers. Solutions or suspensions with conventional pharmaceutical carriers can be used for parenteral administration. For administration by inhalation, the compounds can be in aqueous or partially aqueous solution, which can be used in the form of an aerosol. Agents for topical use can be present, for example, as pharmaceutically acceptable powders, lotions, ointments, creams, gels or as therapeutic systems which contain therapeutically effective amounts of the compounds according to the invention. The dosage required depends on the form of the pharmaceutical agent used, the type of application, the severity of the symptoms and the specific subject (human or animal) being treated. Treatment is usually started at a dose below the optimal dose. The dose is then increased until the optimal effect is achieved for the specified conditions. In general, the compounds according to the invention are best administered in concentrations at which effective effects can be achieved without harmful or disadvantageous effects occurring. They can be administered in a single dose or in multiple doses.

The effectiveness of the compounds according to the invention let yourself determine using various test systems. Further details are given in the given examples.

Methods of Representation of the compounds according to the invention

The synthesis of the compounds according to the invention can be carried out as described in Example 1 below.

Table 1. Growth-inhibitory activity of the substituted 10-phenylimino-9 (10H) -anthracenones against the growth of K562 cells and inhibition of tubulin polymerization in vitro

Table 2 Chemical data of the phenylimino-9 (10H) -anthracenones and the 1,8-dichloro-10-phenylimino-9 (10H) -anthracenones

Examples

The following examples serve the explanation the invention without limiting it as well as better understanding the invention.

example 1

Synthesis of inhibitors tubulin polymerization

Chemical methods

The melting points were determined using a Kofler melting point determination apparatus and are not corrected. Spectra were obtained as follows: ¹ H NMR spectra were recorded on a Varian Gemini 200 (50.3 MHz) spectrometer using tetramethylsilane as the internal standard. Fourier-transformed IR spectra were recorded with a Bio-Rad laboratory type FTS 135 spectrometer and analyzed using WIN-IR Foundation software. Combustion analyzes were carried out in the microanalytical laboratory at the University of Münster using a Heraeus CHN-O rapid analyzer 240. All values are within ± 0.4% of the calculated composition. Mass spectra (EI) were recorded in EI mode using a MAT 44S Finnigan mass spectrometer. All organic solvents have been carefully dried or cleaned before use. Aromatic aldehydes were predominantly commercially available. Analytical thin layer chromatography was carried out on aluminum plates covered with silica gel (Merck Kieselgel 60 F ₂₅₄ , E. Merck, Darmstadt). Column chromatography with silica gel was carried out using Merck silica gel (70-230 mesh).

The structural described here Variations primarily affect the phenyliminopartial structure of the Molecules. The aniline derivatives used as starting compounds were commercial available.

General manufacturing regulations the 10-phenylimino-9 (10H) anthracenone.

Preparation of bromanthrone: 9 (10H) -anthracenone (10 g, 51.5 mmol) is dissolved in 80 ml of chloroform. Then bromine (8.15 g, 2.6 ml, 51 mmol) is slowly added dropwise. There is vigorous development of hydrogen bromide on and after a short time the crude product precipitates crystalline. The mixture is stirred for 30 min at room temperature, then the batch is left to stand in the ice bath for some time, the precipitate is filtered off and the filter cake is washed with about 50 ml of hexane. About 6.0 g of bromanthrone is always obtained as a crude product, which is sufficiently pure for the further reactions. After drying in an oil pump vacuum, the bromanthrone is used without further purification. Preparation of the phenylimino-9 (10H) anthracenones. Bromanthrone (1.0 g, 7.2 mmol) and the respective aniline (7.2 mmol) are suspended in 50 ml of benzene and the mixture is stirred on an oil bath (80 ° C.) for a period of initially 3 hours. After the educt has disappeared (TLC check!), The mixture is allowed to cool to room temperature, the flask is transferred to an ice bath and 150 ml of petroleum ether are added. The resulting precipitate is filtered off after 15 min and washed with hexane or petroleum ether. Most of the product is in the filtrate (DC control!). It is evaporated to dryness in a water jet vacuum. In all cases, the red-brown residue is purified by column chromatography using a short (!) (Cave decomposition!, Not with a 1.8-Cl substitution pattern) silica gel column. When the pure fractions are concentrated, the product crystallizes out after adding hexane and leaving to stand in an ice bath. The phenyliminoanthracenones dissolve in common organic solvents with an intense red color.

The synthesis process described can be carried out analogously starting from 1,8-dichloro-9 (10H) -anthracenone.

10-(3',5'-Dimethoxyphenylimino)-9(10H)-anthracenon

10- (3 ', 5'-Dimethoxyphenylimino) -9 (10H) -anthracenone

FTIR 1656 cm ^-1 ; ¹ H NMR (CDCl ₃ , 200 MHz) δ 8.33-7.38 (in, 8H), 6.26-6.01 (m, 3H), 3.76 (s, 6H); MS m / z (M ⁺ ); Anal. (C ₂₂ H ₁₇ NO ₃ ) C, H, N.

10-(3',4'-Dimethoxyphenylimino)-9(10H)-anthracenon

10- (3 ', 4'-Dimethoxyphenylimino) -9 (10H) -anthracenone

FTIR 1666 cm ^-1 ; ¹ H NMR (CDCl ₃ , 200 MHz) δ 8.40-7.30 (m, 8H), 6.89-6.35 (m, 3H), 3.91 (s, 3H), 3.81 (s, 3H); MS m / z (M ⁺ ); Anal. (C ₂₂ H ₁₇ NO ₃ ) C, H, N.

10-(4'-Methoxyphenylimino)-9(10H)-anthracenon

10- (4'-methoxyphenylimino) -9 (10H) -anthracenone

FTIR 1665 cm ^-1 ; ¹ H NMR (CDCl ₃ , 200 MHz) δ 8.49-7.29 (m, 8H), 6.95, 6.85 (AA ', BB', J _AB = 6.70 Hz), 3.84 (s, 3H); MS m / z (M ⁺ ); Anal. (C ₂₁ H ₁₅ NO ₂ ) C, H, N.

10-(3'-Hydroxy-4'-methoxy-phenylimino)-9(10H)-anthracenone

10- (3'-hydroxy-4'-methoxy-phenylimino) -9 (10H) -anthracenone

FTIR inaccurate cm ^-1 ; ¹ H NMR (CDCl ₃ , 200 MHz) δ 8.47-7.31 (m, 8H), 6.85-6.25 (m, 3H), 5.71 (s, 1H), 3.92 (s, 3H); MS m / z (M ⁺ ); Anal. (C ₂₁ H ₁₅ NO ₃ ) C, H, N.

10-(3',4',5'-Trimethoxyphenylimino)-9(10H)-anthracenon

10- (3 ', 4', 5'-Trimethoxyphenylimino) -9 (10H) -anthracenone

FTIR 2835, 1667 cm ^-1 ; ¹ H NMR (CDCl ₃ , 400 MHz) δ 8.31-7.26 (m, 8H), 6.08 (s, 2H), 3.88 (s, 3H), 3.77 (s, 6H); MS m / z (M ⁺ ); Anal. (C ₂₃ H ₁₉ NO ₄ ) C, H, N.

10-(2',4'-Dimethoxyphenylimino)-9(10H)-anthracenon

10- (2 ', 4'-Dimethoxyphenylimino) -9 (10H) -anthracenone

mp ° C; FTIR cm ^-1 ; 'H NMR (CDCl ₃ , 400 MHz) δ 8.51-7.31 (in, 8H), 6.77-6.50 (m, 3H), 3.84 (s, 3H), 3.58 (s, 3H); MS m / z (M ⁺ ); Anal. (C ₂₂ H ₁₇ NO ₃ ) C, H,

Example 2

Check for growth inhibiting Properties on K562 cells

We tested the newly synthesized compounds for growth-inhibiting properties on the K562 leukemia cell line growing in suspension. ²² The IC ₅₀ value (defined as the effective concentration required for 50% inhibition of customs growth) from 10- (3'-hydroxy-4'-methoxy-phenylimino) -9 (10H) -anthracenone (A 146) in this assay is 0.85 μM ( 2 ).

Human chronic myelogenic K562 leukemia cells were obtained from the German Collection for Microorganisms and Customs Cultures (DSMZ), Braunschweig, and in RPMI 1640 medium (additives: 10% bovine serum (FBS), kanamycin (0.1 mg / ml), penicillin G (100 units / ml), and L-glutamine (30 mg / liter) at 37 ° C and 5% CO ₂ .

The K562 cells were seeded at an inch density of 2 × 10 ⁵ cells / ml in 48-well plates (Costar, Cambridge, MA). Untreated controls were considered 100%. The compounds to be tested were dissolved in DMSO / methanol in a ratio of 1: 1, only the solvent was added to the controls. 5 μL of the dissolved compound were pipetted into each well, so the total volume in the final container was 500 μl. The number of inches was determined using a Model ZM Coulter Counter (Coulter Electronics, Luton, England) after incubation with the compounds to be tested over a period of 48 h.

Table 1 provides an overview of the phenylimino-9 (10H) -anthracenones synthesized and their growth-inhibiting properties (IC ₅₀ values).

The observed antiproliferative effects depend on the substitution pattern. 10- (3'-Hydroxy-4'-methoxy-phenylimino) -9 (10H) -anthracenone A146 showed strong growth-inhibiting properties (IC ₅₀ (K562) 0.85 μM). Compounds 2, 4 and 7 also showed good activity with IC ₅₀ values in the submicromolar range <0.1 μM.

Example 3

Growth-inhibiting properties compared to other tumor inch lines We tested the most active of our compounds (A145, A146, A214, and A215) in vitro for their tumor-inhibiting properties (Table 2) using a propidium iodide-based cytotoxicity assay according to one of Dengler et al. established method. ²³ Again, A145 and A146 proved to be very potent and showed strong growth-inhibiting properties compared to all cell lines used. In particular, the compound was found to be very active against the OVCAR-3 cell line (IC ₅₀ = 0.06 μM).

Example 4

Mikrotubuliassemblierung-turbidimetric assay

Principle of the method

At physiological temperature and in the presence of cofactors (GTP and Mg ²⁺ ions), the microtubuliprolein (MTP) assembles in vitro with the formation of microtubules. This process is reversible. Antagonists of the Assembling, such as Ca ²⁺ ions or even low temperatures (for microtubules from mammalian brain temperatures <10 ° C) require disassembly, while promoters, on the other hand, require stabilization. The kinetics and "steady state" conditions of MT formation and MT disintegration can be followed by time and temperature dependent turbidimetric measurements at 360 nm.

Isolation of MTP and Inhibition of MT assembly

Microtubuliprotein was extracted from pig brain by cycles of temperature-dependent disassembly / reassembly according to a method developed by Shelanski et al. established method. ^27th To increase the yield, the reassembly steps were carried out in the presence of glycerol. A satisfactory purity of the microtubule protein was ensured after two cycles of disassembly / reassembly.

Microtubules for the determination of antimicrotubular effector activities were obtained by in vitro assembly of microtubuliprolein. The microtubule concentration in the assembly studies was 1 mg / ml (determined by the Lowry method using bovine serum albumin). The microtubules assembled in a buffer of the composition 20 mM PIPES (1,4-piperazine diethanesulfonic acid, pK _a 6.8), 80 mM NaCl, 0.5 mM MgCl ₂ , 1 mM EGTA (ethylene glycol bis (2-aminoethylene) tetraacetic acid) after addition of 0.25 mM GTP (guanosine 5'-triphosphate) and heating the samples to 37 ° C. The equilibrium state, recognizable by the no further increasing amount of polymerized tubulin, was reached in the turbidity measurements after an incubation period of 20 min.

Turbidimetric measurement

The turbidimetric measurements were carried out spectrophotometrically at a wavelength of 360 nm. The temperature in the assembly / disassembly cuvette was regulated using a cryostat / thermostat combination. The compounds to be tested were added to a cold MTP solution (MTP concentration 1 mg / ml, temperature 2 to 4 ° C.) in PIPES buffer (for the composition see above) at pH = 6.8. Immediately with the start of the measurement, the temperature was raised to 37 ° C. After a short gap phase, the MTP is assembled to MT. The solution becomes cloudy and reaches an equilibrium state after about 20 minutes. In order to distinguish the turbidity from non-specific protein precipitates, the solution is cooled to 2 ° C. and kept at this temperature for 5 minutes, which requires MT disassembly. The MTP solution becomes transparent again. If the compound to be tested binds or interferes with tubulin, the "steady state" state for inhibitors of the assembly or decreases for substances stabilizing for MT. The IC ₅₀ value indicates the concentration that causes a 50% deviation from the "steady-state" state.

Table 1 shows the IC ₅₀ values of the inhibition of polymerization of pig brain tubulin. Colchicine, nocodazole, podophyllotoxin and vinblastine sulfate are included as reference substances. The compounds A145 (IC ₅₀ = 1.9 μM), A142 (IC ₅₀ = 1.45 μM) and A146 (IC ₅₀ = 0.53 μM) are very potent inhibitors.

The data found prove that the inventive Phenylimino-9 (L0h) anthracenone represent potent cytotoxic agents, their mechanism of action is based on an inhibition of tubulin assembly.

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Claims (12)

A 10-phenylimino-9 (10H) anthracenone of the general formula I,

where the substituents R1-R4 can independently of one another stand for the following substituents: for hydrogen, for alkoxy (C1-C6), for alkyl (C1-C6), for aryloxy, for COOH, for SO ₃ H, for SO ₃ ^- M ⁺ (where M is a cation), for an ester group, for a thioester group, for CN, for an acyl radical, for an aryloxy group, for phenylmethoxy, nitro, hydroxy, amino, imino, mercapto, halogen, for a phosphate ester structure (-OP (O) (OM ⁺ ) ₂ ) or a phosphoramidate (-NP (O) (OM ⁺ ) ₂ ), where M stands for a cation, or (-NP (O) (OR) ₂ ) where it is R is an alkyl radical with up to 8 carbon atoms (the two radicals R can be identical or different, it can be benzyl or aryl radicals). A 1,8-dichloro-10-phenylimino-9 (10H) anthracenone of the general formula II,

where the substituents R1-R4 can independently of one another stand for the following substituents: for hydrogen, for alkoxy (C1-C6), for alkyl (C1-C6), for aryloxy, for COOH, for SO ₃ H, for SO ₃ M ⁺ (where M is a cation), for an ester group, for a thioester group, for CN, for an acyl radical, for an aryloxy group, for phenylmethoxy, nitro, hydroxy, amino, imino, mercapto, halogen, for a phosphate ester structure ( -OP (O) (OM ⁺ ) ₂ ) or a phosphoramidate (-NP (O) (OM ⁺ ) ₂ ), where M stands for a cation, or (-NP (O) (OR) ₂ ) where it is R is an alkyl radical with up to 8 carbon atoms (the two radicals R can be identical or different, it can be benzyl or aryl radicals). A connection according to claim 1 with the structure:

A connection according to claim 2 with the structure:

A connection according to claim 1 with the structure:

A compound according to claim 1 with the structure

A connection according to claim 1 with the structure

A compound according to claim 1 with the structure

A therapeutic composition with a pharmaceutical effective amount of one of the compounds according to claim 1 and a pharmaceutical effective carrier. A pharmaceutical preparation according to claim 9, wherein the pharmaceutically active compound is the compound

is. A therapeutic composition with a pharmaceutical effective amount of one of the compounds according to claim 2 and a pharmaceutical effective carrier. A pharmaceutical preparation according to claim 11, wherein the pharmaceutically active compound is the compound

is.