WO2003068946A1

WO2003068946A1 - Phenalenone derivatives, method for the production thereof and use of the same

Info

Publication number: WO2003068946A1
Application number: PCT/EP2003/001060
Authority: WO
Inventors: Laszlo Vertesy; Michael Kurz; Ziyu Li; Luigi Toti
Original assignee: Aventis Pharma Deutschland Gmbh
Priority date: 2002-02-18
Filing date: 2003-02-04
Publication date: 2003-08-21
Also published as: BR0307733A; PE20030904A1; AU2003210205A1; DE10206849A1; CA2476618A1; MXPA04007918A; AR038524A1; JP2005517710A; EP1478731A1; AU2003210205B2

Abstract

The invention relates to phenalenone derivatives, to the production of the same, whereby the micro-organism Penicillium herquei Bainer & Sartory, DSM 14142, is fermented and the phenalones formed during said fermentation are derivatized, and to the use of said phenalenone derivatives as medicaments, especially for fighting tumour diseases, bacterial infections and/or mycoses and rheumatic diseases.

Description

description

Phenalenone derivatives, process for making and using same.

The present invention relates to phenalenone derivatives, processes for their preparation, and their use as medicaments.

Cancer is a mostly fatal disease of humans and animals, which is caused by the uncontrolled growth of endogenous cells. Cancer is the name given to the formation of malignant tumors (malignancies), neoplasms (tumors or carcinomas) or malignant degeneration and maturation disorders of white blood cells (leukemia, blood cancer). Cancer or tumor cells are formed by the transformation of the body's own cells. The malignancy of the cancerous cell is expressed in the autonomy of growth, that is, in its capacity to be uninhibited and unclassified in the blueprint of the organs and under

Tissue destruction infiltrating to grow. A sure sign of malignancy is the formation of tumor-distant colonies (metastases) after haematogenous or lymphogenic spread of tumor cells. Cancer is one of the most common causes of death in humans and therefore there is a great need for methods and means of curing or treating malignant degeneration.

The possibility of treating malignant tumors includes not only radical removal of the tumor, but also radiological therapy with X-rays, α-, β-, γ-rays, immunotherapy and chemotherapy. Immunotherapy is currently only applicable to a limited extent. Under the

Chemotherapy of tumors is the administration of cytotoxic agents for the treatment of tumors and residual tumor cells after local surgical treatment or radiation. These substances specifically engage in certain processes of cell division, so that tissues with a high proportion of dividing cells, such as the rapidly growing tumor tissue, react more sensitively. Are used alkylating compounds such. For example, cyclophosphamide (The Merck Index, 12th Ed., Page 463), antimetabolites such as methotrexate (The Merck Index, 12th ed. Page 1025), alkaloids such as vincristine (The Merck Index, 12th Ed., Page 1704), and antibiotics such as daunomycin (The Merck Index, 12th ed., Page 479) and adriamycin (The Merck Index, 12th Ed Page 581-582). However, all these agents have major disadvantages due to massive side effects, so that the death of the patients is only delayed, but not averted. In addition, in degenerated (cancer) cells, resistance to the applied means on the current drugs are no longer cytostatic, but toxic because of the side effects. In addition, it has been found that a combined or sequential use of cytostatic drugs exceeds the efficacy of a single cytostatic agent (monotherapy) and thereby is possible that the significant side effects in polychemotherapy do not add up. For all these reasons, new chemotherapeutics are urgently needed and therefore sought worldwide.

Natural products with a phenalenone basic structure have already been described. Phenalen is a condensed, not completely aromatic ring system, which decomposes in the air. Phenalenone is its oxidation product with a carbonyl group in position 1.

The patent application WO 99/60992 generically describes phenalenones which may be substituted in all positions except C1 with hydrogen or C ₁ -C ₄ alkyl, preferably methyl, or C ₁ -C ₄ alkoxy, preferably methoxy, for use as hair colorants.

Japanese Patent Application JP 60199849 describes the phenalenone derivative 2,7,8,9-tetrahydroxy-4-methoxy-5-methyl-phenaleή-1-one, which is effective as a PDE inhibitor and for the treatment of arteriosclerosis, bronchial asthma, diabetes and cancers can be used.

DA Frost & GA Morrison; J. Chem. Soc, Perkin Trans. 1, 20, 2388-2396, 1973 describe the isolation of norxanthoherquine (2,3,4,7,8,9-hexahydroxy-5-methyl-phenalen-1-one) from Penicillium herquei Bainer & Sartory. DHR Barton et al. (Tetrahedron, 6, 48, 1959) describe the isolation of Atrovenetin from Penicillium atrovenetum and from Penicillium herquei Bainer & Sartory. Atroventin is described in Y. Ishikawa et al. (J. Am. Oil Chem. Soc., 68, 666-668, 1991) as an antioxidant, and in WO 00/45165 as an anti-neoplastic cytostatic agent.

N. Narasimhachi et al. (J. Antibiotics, 25, 155, 1972) and J. Simpson (Chem. Soc. Perkin Trans. 1, 1979, 1233-1238) describe tautomeric forms of the compound desoxyherqueinone (2-O-methyl-atrovenetin) derived from Penicillium herquei isolable and show antibiotic activity against Gram-positive organisms.

The invention has for its object to provide alternative phenalenone derivatives that can be used in tumor therapy.

The invention relates to a compound of the formula (I-A)

or a compound of the formula (I-B)

where X is either a group of the formula (Ic)

or the formula (ID) means

(I-D),

and R ¹ and if R ^{2 is} simultaneously 1.0 H, or

2.0 is C ₁ -C ₆ -alkyl, C ₂ -C ₆ -alkenyl or C ₂ -C ₆ -alkynyl, in which alkyl, alkenyl and alkynyl are straight or branched and are optionally monosubstituted or disubstituted by

2.1 -OH,

2.2 O, ^

2.3 -O-Ci-Ce- alkyl, wherein alkyl is straight or branched,

2.4 -OC ₂ -C ₆ -alkenyl, in which alkenyl is straight or branched, 2.5 -aryl,

2.6 -NH-C Cs-alkyl, wherein alkyl is straight or branched,

2.7 - HC ₂ -C ₆ alkenyl wherein alkenyl is straight or branched,

2.8 - H ₂ or

2.9 halogen, in which the substituents 2.1 to 2.9 can also be further substituted by -

CN, amide or oxime functions, ^'

and / or a stereoisomeric form of the compound of the formula (IA) or of the formula (I-B) and / or mixtures of this form in any ratio, and / or a physiologically tolerable salt of the compound of the formula (IA) or (IB). Preferably, R ¹ and R ^{2 are} H or C ₁ -C ₆ -alkyl.

Chirality centers may be in the R or S configuration unless otherwise specified. The invention relates to both the optically pure compounds and stereoisomer mixtures, such as mixtures of enantiomers and Diasteromerengemische, in any ratio.

The compounds according to the invention differ from substances known from the literature by the polarity, the chemical structure, the biological activity or by further physical properties.

The strain Penicillium herquei Bainer & Sartory, DSM 14142, forms on glucose, starch, oatmeal or glycerol-containing nutrient solutions the compounds of the formulas (II-A) and (IIB) called Penilenone, which are summarized below as compounds of the formula (II ),

and atrovenetin of the formulas (III-A) and (III-B), which are referred to collectively below as compounds of the formula (III),

Compounds of formulas (IA) and (IB) for which R ^{1 is} other than H are isomeric forms which are separately isolable and which can be interconverted, for example by cleaving R ¹ , after which R ¹ is H is, and then starting from the other tautomer to the other isomer of the compound of formula (IA) or (IB) is derivatized with R ^{1 is} not H. Hereinafter, the compounds of the formulas (IA) and (IB) are collectively referred to as the compound of the formula (I).

The compounds of formulas (II-A) and (II-B) are tautomers and can not be isolated separately from one another under customary conditions (eg room temperature). Compounds of formulas (II-A) and (II-B) are referred to collectively below as the compound of formula (II).

The compounds of the formulas (III-A) and (III-B) are also tautomers and can not be isolated separately from one another under customary conditions (for example room temperature). Compounds of formulas (III-A) and (III-B) are hereinafter referred to collectively as the compound of formula (III).

Compounds of formulas (IA) and (IB) for which R ¹ is H can be selectively derivatized according to the present invention by reacting the hydroxy groups with alkylating agents in a manner known per se, as described, for example, by Jerry March in Advanced Organic Chemistry, John Wiley & Sons, 4 ^th Edition, 1992. Alkylating agents are for example Diazomethan Dehvate, preferably trimethylsilyldiazomethane. To implement It may be advantageous to selectively introduce protective groups prior to the reaction in a manner known per se. The protective groups are cleaved off after the reaction and then the reaction product is purified.

So far, no selective alkylation of phenalenones has been described to the compounds of the invention. For example, the leads. Reaction of deoxyherqueinone with diazomethane according to Suga et al. (Bull. Chem. Soc., Jpn., 56, 3661-3666, 1983) to the deoxy-quinone dimethyl ether or to the isomeric compound, respectively, are -navrovenetine trimethyl ether. Compounds of formulas (IA) and (IB) for which R ¹ is H can be prepared, for example, by ether cleavage of compounds of formulas (IA) and (IB) for which R ^{1 is} other than H. Ether cleavages can be carried out according to methods known per se, as described, for example, by Jerry March in Advanced Organic Chemistry, John Wiley & Sons, 4 ^th Edition, 1992.

The invention therefore furthermore relates to a compound of the formula (I), which has the formula (II), or a pharmacologically acceptable one

Salt of the compound of formula (II).

The invention therefore further relates to a process for the preparation of a compound of the formula (II), characterized in that

1. the microorganism Penicillium herquei Bainer & Sartory, DSM 14142, or one of its variants or mutants is cultured in an aqueous nutrient medium, 2. a compound of formula (II) is isolated and purified, and

3. The compound of formula (II) is optionally converted into a pharmacologically acceptable salt.

Furthermore, the invention relates to a process for the preparation of a compound of formula (I), characterized in that

1. the microorganism Penicillium herquei Bainer & Sartory, DSM 14142, or one of its variants or mutants is cultivated in an aqueous nutrient medium, 2. a) a compound of the formula (II) is isolated and purified, or b) the compound of the formula (III) is isolated and purified,

3. a) the compound of formula (II) is derivatized to a compound of formula (I), or b) the compound of formula (III) is derivatized to a compound of formula (I), and

4. The compound of formula (I) is optionally converted into a pharmacologically acceptable salt.

The strain Penicillium herquei Bainer & Sartory, DSM 14142, forms the penilenone and the by-products on glucose, starch, oatmeal or glycerol-containing nutrient solutions.

An isolate of Penicillium herquei Bainer & Sartory was deposited with the German Collection of Microorganisms and Cell Cultures GmbH (DSM), Mascheroder Weg 1 B, 38124 Braunschweig, Germany under the rules of the Budapest Treaty on March 6, 2001 under the following number: DSM 14,142th

The fungus Penicillium herquei Bainer & Sartory, DSM 14142 has a gray to bright green substrate mycelium and very little aerial mycelium. Exudates are not formed on malt medium and colors are not eliminated in the medium. The strain, in culture, forms the compact sporangia characteristic of Penicillium, 200-400 x 3.5-4.0 μm, which are rough on the surface. The "Metulae" are relatively short, usually 4-6 10-12 x 3.0-5.0 microns and club-shaped.The phialides are arranged in 6-10 "verticilli", 7-10 x 3.0 microns, ampoule-shaped. The conidia are elliptical to apiculate, 3.5-5.0 x 3.0-3.5 μm, with a smooth cell wall The conidia are formed in parallel chains up to 100 μm long.

The said method comprises the cultivation of Penicillium herquei Bainer & Sartory, DSM 14142, its mutants and / or variants under aerobic conditions in a culture medium containing at least one carbon and nitrogen source, inorganic salts and optionally trace elements. Preferably, the cultivation is carried out at a temperature between 20 ° and 35 ° C and at a pH between 2 and 9.

Instead of the strain DSM 14142 and their mutants and variants can be used, as far as they produce the compounds of the invention. Such mutants can be prepared in a manner known per se by physical means, for example irradiation, such as ultraviolet or X-ray, or chemical mutagens, such as ethyl methanesulfonate (EMS); 2-hydroxy-4-methoxy-benzophenone (MOB) or N-methyl-N'-nitro-N-nitrosoguanidine (MNNG).

Suitable carbon sources for the fermentation are assimilable carbohydrates and sugar alcohols, such as glucose, lactose, sucrose or D-mannitol, as well as carbohydrate-containing natural products, e.g. Malt extract or yeast extract. As nitrogen-containing nutrients are: amino acids, peptides and proteins and their degradation products, such as casein, peptones or tryptones, also meat extracts, yeast extracts, ground seeds, such as corn, wheat, beans, soy, rice or cotton plant, distillation residues of Alcohol production, meat flours or yeast extracts, but also ammonium salts and nitrates, in particular but also synthetically or biosynthetically derived peptides. As inorganic salts, the nutrient solution may contain, for example, chlorides, carbonates, sulfates or phosphates of the alkali metals or alkaline earth metals, iron, zinc, cobalt and manganese.

The formation of the compounds according to the invention proceeds particularly well, e.g. in a nutrient solution containing about 0.05 to 5%, preferably 1 to 2% malt extract; and 0.05 to 3%, preferably 0.05 to 1% yeast extract; 0.2 to 5%, preferably 0.5 to 2% glucose; and 0.5 to 3%, preferably 1.5% to 3% oatmeal contains. The percentages are in each case based on the weight of the entire nutrient solution.

In this nutrient solution, Penicillium herquei Bainer & Sartory, DSM 14142, forms a mixture of the compounds according to the invention. Depending on the composition of the Nutrient solution may vary the quantitative proportion of one or more of the compounds of the invention. In addition, the synthesis of individual compounds can be controlled by the composition of the media, so that a compound is not prepared or in an amount below the detection limit of the microorganism.

The cultivation of the microorganism takes place aerobically, that is, for example, submerged with shaking or stirring in shake flasks or fermenters or on solid medium, optionally with the introduction of air or oxygen. It can be carried out in a temperature range of about 15 to 30 ° C, preferably at about 20 to 30 ° C, especially at 25 to 30 ° C. The pH range should be between 4 and 10, preferably between 6.5 and 7.5. The microorganism is cultivated under these conditions generally over a period of 48 to 720 hours, preferably 72 to 350 hours. Advantageously cultivated in several stages, d. H. one first prepares one or more precultures in a liquid nutrient medium, which are then inoculated into the actual production medium, the main culture, for example in the volume ratio 1: 10-1: 100. The preculture is obtained z. B., by inoculating the mycelium in a nutrient solution and about 20 to 120 hours, preferably 48 to 72 hours to grow. The mycelium can be obtained, for example, by growing the stem for about 1 to 42 days, preferably 21 to 35 days, on a solid or liquid nutrient medium, for example yeast malt agar, oatmeal agar or potato dextrose agar.

The course of fermentation and the screening for mutants and variants which produce the compounds according to the invention can be carried out according to methods known to the person skilled in the art, such as, for example, Example, by testing the biological activity in bioassays or by chromatographic methods such as thin layer chromatography (TLC) or high performance liquid chromatography (HPLC) can be followed.

The fungus Penicillium herquei Bainer & Sartory, DSM 14142, the compounds of the invention by a surface or stand culture on form solid nutrient media. Solid nutrient media are prepared by adding, for example, agar or gelatin to aqueous nutrient media. In addition, it is possible to obtain the compounds according to the invention by fermentation of the fungus Penicillium herquei Bainer & Sartory, DSM 14142, in the submerged process, ie in aqueous slurry. The compounds according to the invention can occur both in the mycelium and in the culture filtrate, usually the major amount is in the cell mass. It is therefore appropriate to separate the fermentation solution by filtration or centrifugation. The filtrate is extracted with an adsorption resin as a solid phase. The mycelium, but also the surface culture is suitably extracted with an organic solvent, for example methanol or propanol-2.

The extractions can be carried out in a wide pH range, but it is expedient to work in a neutral or slightly acidic medium, preferably between pH 3 and pH 7. The extracts can z. B. concentrated in vacuo and dried.

The compounds of formula (II) and formula (III) are substances which are unstable unless special measures are taken during the isolation and purification process. It has been found that the Penilenone can be obtained in very good yields from the cultures of the strain DSM 14142, if 1) during the isolation and purification process is carried out under reducing conditions, eg. B. always in the presence of ascorbic acid, is carried out 2) isolation in an acid medium at a pH of less than 7, preferably in the pH range of 2 to 5) during the cleaning step only mild agents are used 3 ^", such For example, adsorption resins as chromatographic supports, and 4) the presence of amines is excluded throughout the process.

A suitable method of isolating the compounds of the invention is the solvent distribution in a conventional manner. Another method of purification is the chromatography on adsorption resins such. On Diaion® HP-20 (Mitsubishi Casei Corp., Tokyo), Amberlite® XAD 7 (Rohm and Haas, USA), Amberchrom® CG, (Toso Haas, Philadelphia, USA) or the like. In addition, under the circumstances indicated are numerous reverse, phase carrier, z. B. RPβ and RPiβ, as z. B. in the context of high pressure liquid chromatography (HPLC) have become well known. Another possibility for cleaning under the circumstances mentioned in the use of so-called. Normal-phase chromatography supports, such as. As silica gel or Al ₂ 0 ₃ or others in a conventional manner. An alternative isolation method is the use of molecular sieves, such as. B. Fractogel® TSK HW-40, Sephadex® G-25 and others, in a conventional manner.

It is also possible to obtain the compounds of the formula (I) according to the invention after enrichment by crystallization, it being possible, for example, to use organic solvents and their mixtures, anhydrous or with added water.

An additional process for the isolation and purification of the compounds according to the invention consists in the use of anion exchangers, preferably in the pH range from 4 to 7; and cation exchangers, preferably in the pH range from 2 to 5. Particularly suitable for this purpose is the use of buffer solutions to which parts of organic solvents have been added.

However, it is also possible to isolate the compounds of the invention by sublimation and / or to clean.

A particularly advantageous purification method for isolating the compounds according to the invention is crystallization, which is carried out in a manner known per se.

The compounds of the formula (I) according to the invention can be converted into the corresponding pharmacologically acceptable salts by methods known to the person skilled in the art. Among pharmacologically acceptable salts of the invention Compounds according to the invention are understood as meaning both inorganic and organic salts, as described in Remington's Pharmaceutical Sciences (17th Edition, page 1418 [1985]). Salts are in particular alkali, ammonium, alkaline earth salts, salts with physiologically acceptable amines and salts with inorganic or organic acids such. As HCI, HBr, H ₂ S0 ₄ , maleic acid, fumaric acid in question. But there are also complexes with metal ions, such as with calcium, magnesium, zinc, iron or others in question. The compounds of formula (I) have a pronounced tendency to complex ions, preferably cations.

It has surprisingly been found that the compounds of the formula (I) according to the invention have potent cytostatic effects, they are therefore suitable for the therapy and / or prophylaxis of diseases caused by uncontrolled growth of tissue or cells, or tumor diseases. It is particularly noteworthy that the compounds of the invention have no cross-resistance with conventional cytostatics.

It has been found that the compounds of formula (I) inhibit protein kinases. The kinases are among the transferases that transfer phosphate residues from adenosine triphosphate to other substrates. Proteins and enzymes are usually phosphorylated and modified in their activity by the protein kinases on serine, threonine or tyrosine side chains, which has been recognized as a common regulatory principle in metabolism and signal transduction. In the case of cancers, the diseased tissue proliferates unchecked; an intervention in the regulation of kinase-driven proliferation is therefore desirable. The cascade of cell proliferation involves a number of kinases. Several of these kinases are inhibited by the compounds of the invention.

It should also be emphasized an antimicrobial inhibitory effect of the compounds of formula (I) according to the invention on bacteria such as Staphylococcus aureus, Streptomyces murinus and against fungi such as Aspergillus niger, which can cause persistent, life-threatening infectious diseases. The Antimicrobial activity can be detected, for example, by so-called agar-diffusion tests. For example, penilenone on an agar plate containing Streptomyces murinus culture in a solution of 1 mg per mL produces an inhibition zone of 11 mm and in a solution of 0.1 mg per mL a zone of inhibition of 8 mm. The compounds of the formula (I) according to the invention are therefore also suitable for the therapy and / or prophylaxis of bacterial infections and / or fungal diseases.

The compounds of formula (I) can also be used as antioxidants. Antioxidants (oxidation inhibitors) are organic compounds that inhibit or prevent undesirable changes in the substances to be protected due to oxygen effects. Antioxidants are needed, for example, in plastics for protection against aging, in fats for protection against rancidity, in oils against gumming, in odor-deteriorating flavors, in foods, and in medicines. The effect of the antioxidants is usually that they act as a radical scavenger for the occurring during the oxidation of free radicals. The antioxidant activity of atrovenetin (compound of formula (III)) has already been described by Y. Ishikawa et al. J. Am. Oil Chem. Soc. 68, 666-668, 1991. However, microbial antioxidants are often too weak or not well tolerated in their action. There is therefore a great need for new, effective and tolerated antioxidants. The compounds of the formula (I) are highly active antioxidants which significantly exceed the activity of the atrovenetin: while the atrovenetin in solution and in solid form reacts only slowly with atmospheric oxygen (for example in hours), for example the penilenone of the formula ( II) with oxygen within seconds or in a few minutes. However, this increased affinity of penilenone for oxygen is of decisive advantage for very oxidation-sensitive substances.

Another chemical property of the compounds of the invention is the ability to complex with polyvalent, preferably 2- and 3-valent cations, such as Ca ²⁺ , Mg ²⁺ , Zn ²⁺ , Fe ³⁺ . Complexing ability may be beneficial for the preparation of drugs, for example, inhibitors of matrix metallo-proteases (MMP ^'s ) have become known are to bind the zinc of these enzymes. However, other possible uses for diseases are also conceivable, the expression of which manifests itself in an abnormal metal ion concentration in the organism. It is also possible to utilize the complex-forming ability of the compounds according to the invention outside of medicine, for example in water technology, in personal care products, and in polymerization technology (Ullmans Enzyklopadie der Technischen Chemie, 5th edition, A 10, 95-100, 1985-1995 ).

The compounds of the formula (I) according to the invention may likewise be active in the treatment of rheumatic diseases, for example rheumatoid arthritis. The mechanism of action of reducing oxidative stress in rheumatoid arthritis by radical scavengers or antioxidants has been described, for example, by Ostrakhovitch and Afanas (Biochemical Pharmacology, 2001, 743-746).

The present invention accordingly also relates to the use of the compounds of the formula (I) according to the invention as medicaments, in particular for the treatment and / or prophylaxis of tumor diseases, bacterial infections, mycoses, rheumatic diseases and diseases which are manifested by the inhibition of matrix metalloids. Treat proteases.

Furthermore, the present invention relates to a pharmaceutical composition containing at least one of the compounds according to the invention.

Said drug is prepared by mixing at least one compound of formula (I) with one or more physiologically acceptable excipients and placed in a suitable dosage form.

The medicaments according to the invention can be administered enterally (orally), parenterally (intramuscularly or intravenously), rectally or locally (topically). They can be administered in the form of solutions, powders (tablets, capsules including microcapsules), ointments (creams or gel), or suppositories. As physiologically acceptable excipients for such formulations are the pharmaceutical customary liquid or solid fillers and extenders, solvents, emulsifiers, lubricants, flavoring agents, dyes and / or buffer substances in question. As appropriate dosage 0.1 to 1000, preferably 0.2 to 100 mg / kg body weight administered. They are conveniently administered in dosage units containing at least the effective daily amount of the compounds of the invention, for. B. 30 - 3000, preferably 50 - 1000 mg.

The following examples are intended to illustrate the invention without limiting the breadth of the invention in any way.

Example 1 Preparation of a glycerin culture of Penicillium herquei Bainer & Sartory, DSM 14142

30 ml of nutrient solution (malt extract 2.0%, yeast extract 0.2%, glucose 1.0% (NH ₄ ) ₂ HP0 ₄ 0.05%, pH 6.0) in a sterile 100 ml Erlenmeyer flask are prepared with the strain Penicillium Bainer & Sartory, DSM 14142, and incubated for 6 days at 25 ° C and 140 rpm on a rotary shaker. 1.5 ml of this culture are then diluted with 2.5 ml of 80% glycerol and stored at -135 ° C.

Example 2 Preparation of a preculture in the Erlenmeyer flask of Penicillium herquei Bainer & Sartory, DSM 14142

100 ml of nutrient solution (malt extract 2.0%, yeast extract 0.2%, glucose 1.0% (NH ₄ ) ₂ HP0 ₄ 0.05%, pH 6.0) in a sterile 300 ml Erlenmeyer flask are prepared with the strain Penicillium Bainer & Sartory, DSM 14142, and incubated for 4 days at 25 ° C and 140 rpm on a rotary shaker. 2 ml of this preculture are then inoculated for the production of the main cultures.

Example 3 Preparation of a major culture culture of Penicillium herquei Bainer & Sartory, DSM 14142. A sterile 300 ml Erlenmeyer flask containing 100 ml of the following nutrient solution malt extract 2.0%, yeast extract 0.2%, glucose 1, 0% (NH ₄ ) ₂ HP0 ₄ 0.05%, pH 6 is mixed with one on a slant tube (same Nutrient solution, but with 2% agar) grown culture or with 2 ml of a preculture (see Example 2) and incubated on a shaker at 5140 rpm and 25 ° C. The maximum production of one or more compounds of Penilenons invention is reached after about 144 hours. For inoculation of 10 to 200 I fermenters, a 96-144 h old submersed culture (inoculation amount approx. 10%) from the same nutrient solution is sufficient. The conditions for these fermenters are: 0 temperature 25 ° C

Stirrer speed: 200 rpm

Ventilation 15 1 min ^-1

By repeated addition of ethanolic polyol solution foaming can be suppressed. The production maximum is reached after about 96 to 144 hours 5.

Example 4: Isolation of Compounds (II) and (III)

5 liters of culture solution obtained in Example 3 were centrifuged and the 0 cell mass (0.5 liter) was extracted with 2 liters of methanol to which 0.1% ascorbic acid had been added. The clear filtered methanolic phase was concentrated in vacuo to 1 L and applied to a 1 liter, filled with adsorption resin MCI Gel® CHP20P column. Column dimensions: width x height: 7 cm x 27 cm. It was eluted with a solvent gradient of 10% propanol-2 to 90% propanol-2 in 5% 0.1% aqueous ascorbic acid solution. The column effluent (140 mL / min) was collected in fractions of 250 mL each. The penilenone-containing fractions 23 to 26 (mixture of the compounds of the formulas (II-A) and (II-B), comprising compounds of the formula (II)) and the atrovenetin-containing fractions 43 to 51 (mixture of the compounds of the formulas ( III-A) and (III-B), 0 summarizing compounds of the formulas (III) mentioned), which by HPLC-

Analyzes were collected and concentrated in vacuo. The pooled fractions were each concentrated in vacuo and cold stored. From the fractions 23 to 26, the penilenone (260 mg of the compound of the formula (II)) crystallized, and the fractions 43 to 51 gave 1.2 g of atrovenetin (compound of the formula (III)). Under argon inert gas atmosphere, the crystals were filtered off and stored under exclusion of oxygen in the cold.

Example 5: Isolation or purification by HPLC.

Column: Superspher 100 RP-18e®, 250-4, with precolumn, '

Mobile phase: 2 minutes: 5% acetonitrile in 0.1% phosphoric acid, 18 minutes: gradient from 5% to 100% acetonitrile in 0.1%

Phosphoric acid, then 100% acetonitrile uniform.

Flow rate: 1 mL per minute,

Detection by UV absorption at 210 nm.

The retention time of 13.5 minutes for the compound of the formula (II) and 20.5 minutes for the compound of the formula (III) are found.

Example 6: Characterization of the compound of the formula (II).

The physicochemical and spectroscopic properties of penilenone can be summarized as follows:

Appearance: Yellow, in medium polar and polar organic solvents soluble, in water little soluble crystalline substance. The melting point is not determinable because of the decomposition. Stable in a mildly acidic environment under reducing conditions. Under the influence of oxygen, in a neutral environment or in the presence of amines, penilenone turns green.

Molecular formula: Cι ₄ Hιo0 ₆

Molecular weight: 274.23 By ESI + mass spectrometry, a molecular ion 275.2 [M + H] ^{+ was} found, measured under ESI (negative) conditions 273 [M - H] ^~ and 271 [M - 3H] ^~, respectively.

UV maxima: 215 nm, 248 (Sh) nm, 275 (Sh), 389 nm.

Table 1: NMR data - ¹ H and ¹³ C chemical shifts of penilenone of formula (II) in DMSO-d ₆ at 300K (in ppm, numbering for the purpose of NMR evaluation does not correspond to the lUPAC nomenclature).

a) For C3 and C5, no signal is observed in the 13th C spectrum.

Example 7: Obtaining the penilenone monomethyl ether derivatives of the formulas (IV-A) 5 and (IV-B).

40 mg of compound of the formula (II) (penilenone, isolated according to Example 4) were dissolved in 30 ml of tetrahydrofuran and treated with 0.5 ml of 2.0 M (trimethylsilyl) -diazomethane dissolved in hexane [Aldrich, cat. 36,283-2], offset. After an hour, the

10 reaction ended by addition of water and the solvent distilled off in vacuo. The reaction product is then separated on a Nucleosil HD® column (21 mm x 250 mm). The eluent used was a gradient of 10% to 99% acetonitrile in 0.1% acetic acid. The column flow, 20 mL per minute, was fractionally collected. The fractions containing the methylation product

15 were each combined, concentrated in vacuo and crystallized.

Obtained were added 6 mg of Penilenon-dimethyl ether of the formula (IV-A), Molecular Formula: C-ι Hι ₆ 0 _6, molecular weight: 302.29, and 1 mg of Penilenon- dimethyl ether of the formula (IV-B), Molecular Formula:

Molecular weight: 20 302.29.

Penilenone dimethyl ether of the formula (IV-A):

UV maxima: 216 nm, 242 nm, 280 nm (Sh), 387 nm.

25 Table 2: NMR data - ¹ H and ¹³ C chemical shifts δ of the penilenone dimethyl ether of the formula (IV-A) in DMSO-d ₆ at 300K (in ppm, numbering for the purpose of NMR evaluation does not correspond to the lUPAC -Nomenclature).

a) C3 and C5 can not be clearly distinguished.

Penilenone dimethyl ether of the formula (IV-B):

UV maxima: 213 nm, 241 nm and 390 nm.

Table 2: NMR _: data - ¹ H and ¹³ C chemical shifts δ of the penilenone dimethyl ether of the formula (IV-B) in DMSO-d ₆ at 300K (in ppm, numbering for the purpose of NMR evaluation does not correspond to the lUPAC -Nomenclature).

Example 8: Obtaining the Atrovenetin Derivatives (V-A) and (V-B).

100 mg of compound of the formula (III) (atrovenetin, prepared according to Example 4) were dissolved in 5 ml of tetrahydrofuran and treated with 1 ml of 2.0 M (trimethylsilyl) -diazomethane dissolved in hexane [Aldrich, cat. 36,283-2], offset. For ^'15 minutes, the reaction was terminated by addition of water and the solvent distilled off in vacuo. The reaction product is then separated on a Nucleosil AB® column (21 mm x 250 mm). The eluent used was a gradient of 10% to 99% acetonitrile in 10% 0.02% trifluoroacetic acid, which had been adjusted to pH 4.5 with ammonium hydroxide. The column flow, 15 mL per minute, was fractionally collected. The fractions containing the methylation products were each combined, concentrated in vacuo and crystallized.

Were obtained 24 mg of atrovenetin monomethyl ether (VA), empirical formula: C ₂₀ H ₂₀ O ₆ , molecular weight: 356.38, and 10 mg of atrovenetin monomethyl ether (VB), molecular formula C ₂₀ H20O6, molecular weight: 356.38.

Atrovenetin monomethyl ether (V-A):

20th

UV maxima: 218 nm, 260 nm (Sh), 394 nm.

Table 3: NMR data - ¹ H and ¹³ C chemical shifts δ of the atrovenetin monomethyl ether (VA) in DMSO-d ₆ at 300 K (in ppm, numbering for purpose 25 of the NMR evaluation does not correspond to the lUPAC nomenclature) ,

Atrovenetin monomethyl ether (V-B):

UV maxima: 222 nm, 282 nm, 385 nm.

Table 4: NMR data - ¹ H and ¹³ C chemical shifts δ of the atrovenetin monomethyl ether (VB) in DMSO-d ₆ at 300 K (in ppm, numbering for the purpose of NMR analysis does not correspond to the lUPAC nomenclature).

Claims

Patent claims:

1. Compound of formula (l-A)

or a compound of the formula (l-B)

where X is either a group of the formula (l-C)

or the formula (ld) means,

and R-! and if present R ² simultaneously 1. OH, or

2.0 means Ci-Ce-alkyl, C ₂ -C ₆ -alkenyl or C ₂ -C ₆ -alkynyl, in which alkyl, alkenyl and alkynyl are straight or branched and optionally mono- or disubstituted 5. are by:

2.1 -OH,

2.2 =0,

2.3 -O-Cι-C ₆ -alkyl, in which alkyl is straight or branched,

2.4 -OC ₂ -C ₆ -alkenyl, in which alkenyl is straight or branched, 0 2.5 -aryl,

2.6 -NH-C C _ό - alkyl, in which alkyl is straight or branched,

2.7 - HC ₂ -C ₆ - alkenyl, in which alkenyl is straight or branched,

2.8 -NH ₂ or

2.9 halogen, 5 in which the substituents 2.1 to 2.9 can also be further substituted by -

CN, -amide or -oxime functions,

and/or a stereoisomeric form of the compound of the formula (l-A) or the formula (l-B) and/or mixtures of this form in any ratio, and/or a physiologically compatible salt of the compound of the formula (l-A) or (l-B) .

2. A compound of formula (lA) or (lB) according to claim 1, wherein Rl and R ² are H or CrC-β-alkyl.

5 3. Compound of the formula (1A) or (1B) according to one or more of claims 1 to 2, characterized in that it has the formula (II-A)

or the formula (ll-B)

as well as their physiologically tolerated salts.

4. A process for producing a compound of the formula (1-A) or (1-B) according to one or more of claims 1 and 2, characterized in that 1. the microorganism Penicillium herquei Bainer & Sartory, DSM 14142, or one of its variants or mutants is cultivated in an aqueous nutrient medium, 2. a) a compound of the formula (II-A)

or the formula (II-B)

is isolated and purified, or b) a compound of the formula (III-A)

or the formula (III-B)

is isolated and purified, 3. a) the compound of the formula (II-A) or (II-B) is derivatized to a compound of the formula (IA) or (IB), or b) the compound of the formula (III-A) or (III-B) is derivatized to a compound of the formula (lA) or (lB), and 4. the compound of the formula (lA) or (lB) optionally in a pharmacological compatible salt is transferred.

5. A process for preparing a compound of the formula (II-A) or (II-B) according to claim 3, characterized in that

1. the microorganism Penicillium herquei Bainer & Sartory, DSM 14142, or one of its variants or mutants, is cultivated in an aqueous nutrient medium,

2. a compound of the formula (II-A) or (II-B) is isolated and purified, and

3. the compound of the formula (II-A) or (II-B) is optionally converted into a pharmacologically acceptable salt.

6. The method according to claim 4, wherein the derivatization takes place using an alkylating agent.

7. The method according to claim 6, wherein the alkylating agent is a diazomethane-r derivative.

8. Use of a compound according to one or more of claims 1 to 3 for producing a medicament.

9. Use of a compound according to claim 8 for producing a medicament for the treatment and prophylaxis of tumor diseases.

10. Use of a compound according to claim 8 for producing a medicament for the treatment and prophylaxis of bacterial infectious diseases and/or mycoses

11. Use of a compound according to claim 8 for producing a medicament for the treatment and prophylaxis of rheumatoid diseases.

12. Use of a compound according to claim 8 for the production of a medicament against diseases that can be treated by inhibiting matrix metallo-proteinases.

13. Use of a compound according to one or more of claims 1 to 3 as an antioxidant.

14. Medicaments containing at least one compound according to one or more of claims 1 to 3 and one or more physiologically acceptable excipients.

15. Strain Penicillium herquei Bainer & Sartory, DSM 14142.