WO2003055843A1 - Agaricoglycerides and analogs - Google Patents

Abstract

The invention relates to agaricoglycerides and analogs, methods for the production thereof, in addition to the use thereof for the production of medicaments for treating and/or the prophylaxis of illnesses, especially painful conditions.

Description

Agaricoglycerides and analogs

The invention relates to agaricoglycerides and analogs, ner processes for their manufacture and their use for the manufacture of medicaments for treatment and / or

Prophylaxis of diseases, especially painful conditions.

In the past few years, chlorine-containing aromatic secondary substances have been increasingly isolated from cultures of Basidiomycetes, e.g. from the genera Hypholoma and Stropharia, which are placed in the family Strophariaceae. Another family of Basidiomycetes, the Agaricaceae, contains fungi of the genus Agaricus. The polychlorinated aromatic antibiotic drosophilin A, which had previously also been found in a species of the genus Drosophila (syn. Psathyrella) belonging to the Coprinaceae, was isolated a long time ago from one of its representatives, the cultivated mushroom Ägaricus bisporus. The article by E. de Jong & J.A. provides a comprehensive overview of aromatic organochlorine compounds identified to date from this group of fungi. Field, Annu. Rev. Microbiol, 1997, 51, 375-414.

In the context of the present invention it was possible to obtain new agaricoglycerides from fungi of the genus Agaricus. These substances surprisingly inhibit the Νeurolysin and are therefore suitable for the treatment of pain and rheumatic diseases.

Oleurolysin (EC 3.4.24.16) is a Zn metalloprotease that inactivates a number of biologically active peptides such as Νeurotensin and dynorphin A by cleavage (Shrimpton and Smith J Peptide Sci. 2000, 6, 251-263; Vincent et al. Brain Res. 1996, 709, 51-58). As a δ-opioid receptor agonist, dynorphin A has a strong analgesic effect, and when applied centrally, neurotensin also has an analgesic effect (Tyler et al. Brain Res. 1998, 792, 246-252). Neurolysin inhibitors should therefore increase the analgesic effects of neurotensin and dynorphin A by inhibiting the inactivation of these peptides.

Certain salicylic acid alkane polyol esters, such as, for example, salicylic acid glycerol esters, which are synthesized or from molluscs of the genera Kelletia and

Buccinulum have been isolated, i.a. in Experientia 1996, 52, 812-817 as HIN-1 reverse transcriptase inhibitors with antiviral activity.

Other non-medical applications are known for certain substituted benzoic acid glycerol esters: as additives for polymeric materials for medical devices, e.g. Hoses and containers (EP-A-0 576 868); as a tanning agent (WO 92/18457); as additives for light-sensitive photomaterials and light-sensitive compositions (EP-A-0 268 496, DE-A 29 01 630 and DE-A 29 11 286); as UV stabilizers of polymers (US-A-3,888,823, US-A-3, 206,431 and US-A-4,619,957); as starting materials for the production of an anticorrosive coating (DE-A-3441 043).

Non-medical applications for certain substituted benzoic acid diol esters are described: as starting materials for the production of polyesters (US-A-4,065,432, JP-A 60 094 942, JP-A 60 019 751, JP-A 51 033 187); as starting materials for the production of polycarbonate resins for electrophotographic photoconductors (US-B 6,187,492); as additives for a positive photoresist (DE-A 43 04 098); as additives for electrophotographic toners (JP-A 10 161 351).

The synthesis of benzoic acid diol esters is claimed in WO 92/18459.

The present invention relates to compounds of the formula

for a rest of the formula

R, R, R, R, R and R independently of one another are radicals selected from the group consisting of hydrogen, halogen, cyano, C 1 -C ₄ -alkyl and C 1 -C ₄ -alkoxy,

wherein at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} halogen or

Cyano means

R, R and R independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy, acetoxy, amino, carboxyl, halogen, cyano and C 1 -C ₄ -alkyl,

where at least one of the radicals R ² , R ⁵ and R ^{8 is} hydroxy, acetoxy, amino or carboxyl,

and 9 R where R, R and R are each in the meta or para position of the benzoyl radicals,

R ^{10 represents} hydrogen or -CC ₆ alkyl,

or

E stands for -CR ₁₁ R ₁₂ -

R represents halogen or cyano,

R represents hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another represent radicals selected from the group consisting of hydrogen, halogen and cyano,

R ^{8 represents} hydrogen, hydroxy or acetoxy,

and

where R ² and R ^{8 are} each in the meta or para position of the benzoyl radicals,

R ^{11 represents} hydrogen or - alkyl,

R ^{12 represents} C ₆ -C ₁₀ aryl or 5- to 10-membered heteroaryl,

where Ce-Cio-aryl and 5- to 10-membered heteroaryl with up to 3 substituents selected independently from the group halogen, cyano, trifluoromethyl, Cr alkyl and C ₁ -C ₆ alkoxy may be substituted, or

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a 4- to 6-membered cycloalkyl ring,

or

E for a remainder of the formula

or stands

R ^{1 represents} halogen or cyano,

R ^{2 represents} hydroxy or acetoxy,

R, R and R independently of one another for radicals selected from the group

Are hydrogen, halogen, -CC alkyl and cyano,

R ^{8 represents} hydrogen, hydroxy or acetoxy,

and

where R ² and R ^{8 are} each in the meta or para position of the benzoyl radicals,

R ^{10 represents} hydrogen or -CC ₆ alkyl,

R ^{13 represents} phenyl, where phenyl can be substituted with up to 3 substituents independently of one another selected from the group consisting of halogen, cyano, trifluoromethyl, -Ce-alkyl, -CC ₆ alkoxy and -CO ₂ R ¹⁵ ,

in which

R ^{15 represents} Ci-Cβ-Al yl and benzyl.

The present invention further relates to the use of compounds of the formula (I)

in which

stands for - (CH ₂ ) „- or -CR ₁₆ Rι ₇ -,

R represents halogen or cyano,

R ^{2 represents} hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another for radicals selected from the group

Are hydrogen, halogen and cyano,

R ^{8 represents} hydrogen, hydroxy or acetoxy,

and

where R ² and R ^{8 are} each in the meta or para position of the benzoyl radicals,

n stands for 0 to 3,

R ^{16 represents} hydrogen, -CC ₆ -alkyl or C ₃ -C ₈ -cycloalkyl, and

R ^{17 represents} C ₁ -C ₆ -alkyl or C ₃ -C ₈ -cycloalkyl,

C ₁ -C ₆ -alkyl may optionally be substituted by phenyl,

for the manufacture of a drug.

The compounds according to the invention can also be in the form of their salts, solvates or

Solvates of the salts are present.

The compounds according to the invention can exist in stereoisomeric forms (enantiomers, diastereomers). The invention therefore relates both to the enantiomers or diastereomers and to their respective mixtures. These enantiomer and diastereomer mixtures can be separated into the stereoisomerically uniform constituents in a known manner.

In the context of the invention, preferred salts are physiologically acceptable salts of the compounds according to the invention.

Physiologically acceptable salts of the compounds according to the invention can be acid addition salts of the compounds with mineral acids, carboxylic acids or sulfonic acids. For example, particular preference is given to Salts with hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, ehanesulfonic acid,

Toluenesulfonic acid, benzenesulfonic acid, naphthalenedisulfonic acid, acetic acid, propionic acid, lactic acid, tartaric acid, citric acid, fumaric acid, maleic acid or benzoic acid.

Physiologically and questionable salts can also be salts with conventional bases, such as, for example, alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example Calcium or magnesium salts) or ammonium salts, derived from ammonia or organic amines such as, for example, diethylamine, triethylamine, ethyldiisopropylamine, procaine, dibenzylamine, N-methylmorpholine, dmydroabieylamine, 1-ephenamine or methylpiperidine.

In the context of the invention, solvates are those forms of the compounds which form a complex in the solid or liquid state by coordination with solvent molecules. Hydrates are a special form of solvate, in which coordination takes place with water.

In the context of the present invention, the substituents generally have the following meaning:

GCRG-alky! stand for a straight-chain or branched alkyl radical having 1 to 6, preferably 1 to 4, particularly preferably having 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, tert-butyl, n-pentyl and n-hexyl.

C 1 -C ₆ -alkoxy represents a straight-chain or branched alkoxy radical having 1 to 6, preferably 1 to 4, carbon atoms. Non-limiting examples include methoxy,

Ethoxy, n-propoxy, isopropoxy and tert-butoxy.

Cfi-Cin-Aryl stands for an aromatic radical with 6 to 10 carbon atoms. Preferred aryl radicals are phenyl and naphthyl.

5- to 10-membered heteroaryl stands for 5- to 10-membered aromatic rings containing heteroatoms with at least one aromatic ring, which can contain 1 to 4 heteroatoms, which can be selected from O, S and N. Heteroaryl can in turn be added via C or N may be substituted. Non-limiting examples include pyridyl, furyl, thienyl, pyrrolyl, imidazolyl, pyrazolyl, pyrazinyl, pyrimidinyl, pyridazinyl, Indolicenyl, indolyl, benzo [b] thienyl, benzo [b] furyl, indazolyl, quinolyl, isoquinolyl, naphthyridinyl and quinazolinyl.

4- to 6-membered cycloalkyl ring represents a cycloalkyl group with 4 to 6 carbon atoms. Non-limiting examples include cyclobutyl, cyclopentyl and

Cyclohexyl.

Halogen stands for fluorine, chlorine, bromine and iodine. Fluorine, chlorine and bromine are preferred, particularly preferably fluorine and chlorine.

If radicals in the compounds according to the invention are optionally substituted, the radicals, unless otherwise specified, can be substituted one or more times in the same or different manner. A substitution with up to three identical or different substituents is preferred. Substitution with a substituent is very particularly preferred.

For the purposes of the invention, the meta (m) and para (p) positions of the benzoyl radicals are defined as follows:

The present invention further relates to compounds of the formula (I)

in which

E for a remainder of the formula

stands,

R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 are} independently selected for residues from the

Are hydrogen, halogen, cyano, C ₁ -C ₄ -alkyl and -CC ₄ -alkoxy,

where at least one of the radicals R, R, R, R, R and R is halogen or cyano,

R ² , R ⁵ and R ⁸ independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy, acetoxy, amino, carboxyl, halogen, cyano and C 1 -C ₄ -alkyl,

where at least one of the radicals R, R and R is hydroxyl, acetoxy, amino or carboxyl,

and

where R, R and R are each in the meta or para position of the benzoyl radicals,

and

R ^{10 represents} hydrogen or -CC ₆ alkyl. Compounds of the formula (I) in which

for a rest of the formula

stands,

R ^{1 represents} halogen or cyano,

R> 2 represents hydroxy, acetoxy, amino or carboxyl,

R ³ , R ⁴ , R ⁶ , R ⁷ and R ⁹ independently of one another for radicals selected from the group

Are hydrogen, halogen, cyano, C 1 -C ₄ -alkyl and C 1 -C ₄ -alkoxy,

R ⁵ and R ⁸ independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy, acetoxy, amino, carboxyl, halogen, cyano and C 1 -C ₄ -alkyl,

and

where R ² , R ⁵ and R ^{8 are} each in the meta or para position of the benzoyl radicals,

and

R ^{iυ represents} hydrogen or -CC ₆ alkyl. Also preferred are compounds of formula (I) in which

E for a remainder of the formula

stands,

R represents halogen or cyano,

R ^{5 represents} hydroxy, acetoxy, amino or carboxyl,

R ¹ , R ³ , R ⁶ , R ⁷ and R ⁹ independently of one another represent radicals selected from the group consisting of hydrogen, halogen, cyano, C ₁ -C ₄ alkyl and C ₁ -C ₄ alkoxy,

9 p

R and R independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy, acetoxy, amino, carboxyl, halogen, cyano and C 1 -C ₄ -alkyl,

and

where R> 2, R and R are each in the meta or para position of the benzoyl radicals,

and

R, 1 ¹ 0 ^{U represents} hydrogen or -CC ₆ alkyl. Also preferred are compounds of formula (I) in which

E for a remainder of the formula

stands,

R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 are} independently selected for residues from the

Are hydrogen and halogen,

where at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} halogen,

and R .2, _D R5, _D R8 "and" dj _π R10 have the meanings given above.

Compounds of the formula (I) in which

E for a remainder of the formula

stands,

R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 are} independently selected from the

Group hydrogen and chlorine, where at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} chlorine,

and

have the meanings given above.

Also preferred are compounds of formula (I) in which

for a rest of the formula

stands,

R ² , R ⁵ and R ⁸ independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy, acetoxy, amino and carboxyl,

wherein at least one of the radicals R ² , R ⁵ and R ^{8 is} hydroxy, acetoxy, amino or carboxyl, and

9 * ϊ R where R, R and R are each in the meta or para position of the benzoyl radicals,

and R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ , R ⁹ and R ^{10 have} the meanings given above.

Compounds of the formula (I) in which

E for a remainder of the formula

stands,

R ² , R ⁵ and R ^{8 are} each in the para position of the benzoyl radicals,

and R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ , R ⁹ and R ^{10 have} the meanings given above.

Compounds of the formula (I) in which

E for a remainder of the formula

stands,

R ² , R ⁵ and R ⁸ are selected independently of one another from the group hydrogen, hydroxy and acetoxy,

wherein at least one of the radicals R ² , R ⁵ and R ^{8 is} hydroxy or acetoxy, and

where R ² , R ⁵ and R are each in the para position of the benzoyl radicals, and R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ , R ⁹ and R ^{10 have} the meanings given above.

Also preferred are compounds of formula (I) in which

for a rest of the formula

stands,

R ^{10 represents} hydrogen,

and R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ and R ^{9 have} the meanings given above.

Combinations of two or more of the preferred ranges mentioned above are very particularly preferred.

Compounds of the formula (I) are also very particularly preferred,

in which

E for a remainder of the formula

stands,

R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ⁹ independently of one another represent hydrogen or chlorine,

where at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} chlorine,

R, R and R independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy and acetoxy,

where at least one of the radicals R ² , R ⁵ and R ^{8 is} hydroxy or acetoxy,

and

where R ² , R ⁵ and R ^{8 are} each in the para position of the benzoyl radicals,

and

R, 1 ^ι 0 ^υ stands for hydrogen.

Compounds of the formula (I) are also very particularly preferred,

in which

E for a remainder of the formula

stands,

R ^{1 represents} chlorine,

R ^{2 represents} hydroxy or acetoxy,

R ³ , R ⁴ , R ⁶ , R ⁷ and R ⁹ independently of one another represent hydrogen or chlorine,

R ⁵ and R ⁸ independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy and acetoxy,

and

where R, 2, R r> 5 and R are each in the para position of the benzoyl radicals,

and

R .1 ^ι 0 ^υ stands for hydrogen.

Compounds of the formula (I) are also very particularly preferred,

in which

for a rest of the formula

stands,

R ^{4 represents} chlorine,

R ^{5 represents} hydroxy or acetoxy,

R ¹ , R ³ , R ⁶ , R ⁷ and R ⁹ independently of one another represent hydrogen or chlorine,

R ² and R ⁸ independently of one another represent radicals selected from the group consisting of hydrogen, hydroxy and acetoxy,

and

where R> 2, R and R are each in the para position of the benzoyl radicals,

and

R ^{ιυ stands} for hydrogen.

The present invention further relates to compounds of the formula (I)

in which

E stands for -CR ₁ 1R12-, R ^{1 represents} halogen or cyano,

R represents hydroxy or acetoxy,

R, R and R independently of one another for radicals selected from the group

Are hydrogen, halogen and cyano,

R ^{8 represents} hydrogen, hydroxy or acetoxy,

and

where R ² and R ^{8 are} each in the meta or para position of the benzoyl radicals,

R ^{11 represents} hydrogen or -CC ₄ alkyl,

R ^{12 represents} C ₆ -Cιo-aryl or 5- to 10-membered heteroaryl,

where Ce-Cio-aryl and 5- to 10-membered heteroaryl with up to 3 substituents independently selected from the group halogen, cyano, trifluoromethyl, C ₁ -C ₆ -alkyl and C ₁ -C ₆ -alkoxy can be substituted,

or

R ¹¹ and R ¹² together with the carbon atom to which they are attached form a 4- to 6-membered cycloalkyl ring.

Compounds of the formula (I) are very particularly preferred

in which

stands for -CR11R ₁ 2-, R ¹ , R ³ , R ⁷ and R ^{9 represent} chlorine,

where R ¹ , R ³ , R ⁷ and R ^{9 are} each in the meta position of the benzoyl radicals,

R ² and R ^{8 represent} hydroxy,

where R ² and R ^{8 are} each in the para position of the benzoyl radicals,

R ^{n represents} hydrogen or methyl,

and

R ^{12 represents} phenyl, pyridyl or quinolinyl,

where phenyl can be substituted once with methoxy.

The present invention further relates to compounds of the formula (I)

in which

E for a remainder of the formula

or stands

R ^{1 represents} halogen or cyano,

R ^{2 represents} hydroxy or acetoxy, R, R and R independently of one another are radicals selected from the group consisting of hydrogen, halogen, C 1 -C ₄ -alkyl and cyano,

R represents hydrogen, hydroxy or acetoxy,

and

9 R where R and R are each in the meta or para position of the benzoyl radicals,

R »1 ^ι 0 ^{υ represents} hydrogen or CC ₆ alkyl,

and

R ^{13 represents} phenyl,

where phenyl with up to 3 substituents independently selected from the group halogen, cyano, trifluoromethyl, C ₁ -C ₆ alkyl, C ₁ -C ₆ alkoxy and -CO ₂ R ¹⁵ may be substituted,

where R ^{15 is} -Ce-alkyl and benzyl.

Compounds of the formula (I) are very particularly preferred

in which

E for a remainder of the formula

stands, R ¹ , R ³ , R ⁷ and R ^{9 represent} chlorine,

where R ¹ , R ³ , R ⁷ and R ^{9 are} each in the meta position of the benzoyl radicals,

R 9 and R R represent hydroxy,

where R ² and R ^{8 are} each in the para position of the benzoyl radicals,

R ^{14 represents} hydrogen,

and

R ^{13 represents} phenyl,

where phenyl can be substituted with up to 3 substituents independently selected from the group consisting of chlorine, fluorine, bromine, trifluoromethyl, C 1 -C ₄ alkyl, methoxy and -CO ₂ benzyl.

The present invention further relates to the following compounds

2 - [(5-Chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] -l - {[(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] methyl} ethyl-4 - (benzyloxy) -5-chloro-2,3-difluorobenzoate

2,3-bis [(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] propyl 5-chloro-2,3-difluoro-4-hydroxybenzoate

2 - [(3,5-dichloro-4-hydroxybenzoyl) amino] -l - {[(3,5-dichloro-4-hydroxybenzoyl) oxy] - methyl} ethyl-3,5-dichloro-4-hydroxybenzoate

and their salts, solvates and solvates of the salts.

The present invention further preferably relates to the use of compounds of the formula (I)

in which

stands for - (CH ₂ ) _n - or -CR ₁₆ Rι ₇ -, R ^{1 represents} chlorine,

R ^{2 represents} hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another represent hydrogen or chlorine,

R »» stands for hydrogen, hydroxy or acetoxy,

and

9 R where R and R are each in the para position of the benzoyl radicals,

n represents 1 to 3,

R ^{16 represents} hydrogen or methyl,

and

R ¹⁷ represents C 1 -C ₄ -alkyl,

where dC ₄ alkyl can optionally be substituted with phenyl,

for the manufacture of a drug.

The invention further relates to processes for the preparation of the compounds of

Formula (I) characterized in that

by procedure [A] Compounds of the formula

in which

R, ι to R, R to R and R 10 have the meanings given above,

with compounds of the formula

in which

R, R and R have the meanings given above,

and

X ^{1 represents} halogen, preferably bromine or chlorine, or hydroxy,

to compounds of the formula

in which

R, 1 to R, 10 have the meanings given above,

be implemented

or

[B] Compounds of the formula

in which

R, ι to R have the meanings given above,

E for -CR _! ιRι ₂ -, - (CH ₂ ) _n -, -CR ₁₆ R ₁₇ -

or a residue of the formula

stands, and

n, R »10 ι bi • s ι R- 13, _π Rl ₆ - and" R "17 have the meanings given above,

with preferably one equivalent of a compound of the formula

in which

R »7, _π R8. and, τR> 9 have the meanings given above,

and

X rl represents halogen, preferably bromine or chlorine, or hydroxy,

to compounds of the formula

in which

E for -CR11R12-, - (CH ₂ ) _n -, -CRiβRi? - or a residue of the formula

stands,

and

n, R to R, R to R, R and R have the meanings given above,

be implemented

or

[C] Compounds of the formula (II) with compounds of the formula

in which

R has the meanings given above,

to compounds of the formula

in which

i ^ 7 1 π i "

R to R, R to R and R have the meanings given above,

be implemented

and the resulting compounds (la), (Ib) and (Ic) are optionally reacted with the corresponding (a) solvents and / or (b) bases or acids to give their solvates, salts or solvates of the salts.

Formula (I) comprises the compounds (Ia), (Ib) and (Ic).

9 rows

The hydroxyl group of the radicals R, R and R optionally carries a protective group, such as. B. benzyl, which can be split off by methods known to those skilled in the art.

Some of the compounds of the formula (II) are known or can be prepared by using compounds of the formula

in which

R ¹ , R ² , R ³ and R ^{10 have} the meanings given above,

with preferably one equivalent of a compound of formula (V). Some of the compounds of formula (VII) are known or can be prepared by using compounds of formula

in which

R ^{10 has} the meanings given above,

with preferably one equivalent of a compound of the formula

in which

1 9

R, R and R have the meanings given above,

and

X ^{3 represents} halogen, preferably bromine or chlorine, or hydroxy,

implements.

The compounds of formula (IV) are known or can be prepared by using compounds of formula

in which

E for -CR „R ₁₂ -, - (CH ₂ ) _n -, -CR ₁₆ R ₁₇ -

or a residue of the formula

stands, and

n, R ¹⁰ to R ¹³ , R ¹⁶ and R ^{7 have} the meanings given above,

with preferably one equivalent of a compound of formula (IX).

The compounds of the formula (X) are known (for E = - (CH ₂ ) _n - or -CRι ₆ Rι -) or can be prepared, for example (for E = -CRπRι ₂ -) according to synthesis scheme 1 (see for cycloalkyl ring: J Am. Chem. Soc. 1960, 82, 2932-2936; for quinoline: Tetrahedron Asymmetry 1998, 9, 2481-2492; for aryl and heteroaryl: J Chem. Soc. 1925, 127, 2502-2507; for alkylaryl: Can. J. Chem. 1979, 57, 1025-1032; J Org. Chem. 1984, 49, 4226),

Synthesis scheme 1:

or the compounds of formula (X) can be prepared by using compounds of formula

in which

R, 10 has the meanings given above,

with preferably one equivalent of a compound of the formula

in which

R ^{13 has} the meanings given above, and X ^{4 represents} halogen, preferably bromine or chlorine, hydroxy or methoxy,

to compounds of the formula

in which

R ¹⁰ and R ^{13 have} the meanings given above,

implements.

Formula (X) includes compounds (Xa).

For the syntheses [(II) + (III) -> (la)], [(IV) + (V) -> (Ib)], [(VII) + (V) -> (II)], [( VIII) + <TX) -> (VII)], [(X) + (IX) -> (IV)] and [(XI) + (XII) -> (Xa)] applies:

If, X ¹ , X ² , X ³ and X ^{4 represent} halogen,

is reacted, generally in inert solvents, if appropriate in the presence of a base and of 4-dimethylaminopyridine, preferably in a temperature range from 0 ° C. to 50 ° C. at atmospheric pressure.

Inert solvents are, for example, halogenated hydrocarbons such as methylene chloride, trichloromethane, carbon tetrachloride, trichloroethane, tetrachloroethane, 1,2-dichloroethane or trichlorethylene, ethers such as diethyl ether, methyl tert-butyl ether,

Dioxane, tetrahydrofuran, glycol dimethyl acid or diethylene glycol dimethyl ether, hydrocarbons such as benzene, xylene, toluene, hexane, cyclohexane or petroleum ions, nitroalkanes such as nitromethane, carboxylic acid esters such as ethyl acetate, ketones such as acetone or 2-butanone, carboxylic acid amides such as dimethylformamide or dimethylacetamide, alkyl sulfoxides such as dimethyl sulfoxide, aliphatic nitriles such as acetonitrile or heteroaromatics such as pyridine, preference is given to dioxane or tetrahydrochloride, tetrahydrofuran, for tetrahydrofuran.

Bases are, for example, alkali metal hydroxides such as sodium or potassium hydroxide, or alkali metal carbonates such as cesium carbonate, sodium or potassium carbonate, or amides such as lithium diisopropylamide, or organic amines such as DBU, triethylamine or diisopropylethylamine, preferably diisopropylethylamine or triethylamine.

If, X ¹ , X ² , X ³ and X ^{4 represent} hydroxy,

is generally used in inert solvents, in the presence of customary condensing agents, if appropriate in the presence of a base, preferably in one

Temperature range from room temperature to 50 ° C at normal pressure.

Preferred inert solvents are tetrahydrofuran, dimethylformamide, 1,2-dichloroethane or methylene chloride. ^•

Typical condensing agents are, for example, carbodiimides such as, for example, N, N'-diethyl, N, N, '- dipropyl, N, N'-diisopropyl, N, N'-dicyclohexylcarbodiimide, N- (3-dimethylaminoisopropyl) -N'-ethylcarbodiimide hydrochloride (EDC), N-cyclohexyl-carbodiimide-N'-propyloxymethyl-polystyrene (PS-carbodiimide) or carbonyl compounds such as carbonyldiimidazole, or 1,2-oxazolium compounds such as 2-ethyl-5-phenyl- 1, 2-oxazolium-3 sulfate or 2-tert. -Butyl-5-methyl-isoxazolium perchlorate, or acylamino compounds such as 2-ethoxy-l-ethoxycarbonyl-l, 2-dihydroquinoline, or propanephosphonic anhydride, or isobutylchloroformate, or bis (2-oxo-3-oxazolidinyl) phosphoryloxy or benzotriazoloxy -tri (dimethylamino) phosphonium hexafluorophosphate, or O- (benzotriazol-l-yl) -N, N, N ', N'-tetra-methyluronium hexafluorophosphate (HBTU), 2- (2-oxo-1 - (2H) -pyridyl) - 1, 1, 3,3 -tetramethyl- uronium tetrafluoroborate (TPTU) or O- (7-azabenzotriazol-l-yl) -N, N, N ', N'-tetra-methyl-uronium hexafluorophosphate (HATU), or 1-hydroxybenztriazole (HOBt), or benzotriazole-1 -yloxytris (dimethylamino) -phosphonium hexafluorophosphate (BOP), or mixtures of these.

Bases are, for example, alkali carbonates, e.g. Sodium or potassium carbonate, or hydrogen carbonate, or organic bases such as trialkylamines e.g. Triethylamine, N-methylmorpholine, N-methylpiperidine, 4-dimethylaminopyridine or diisopropylethylamine.

The combination of N- (3-dimethylaminoisopropyl) -N'-ethylcarbodiimide hydrochloride (EDC), 1-hydroxybenzotriazole (HOBt) and triethylamine in dimethylformamide or carbonyldiimidazole in 1,2-dichloroethane is particularly preferred.

If X ^{4 is} methoxy,

is reacted without solvent, preferably in a temperature range from 100 ° C to 120 ° C at normal pressure.

The synthesis [(II) + (VI) -> (Ic)] takes place under Mitsunobu reaction conditions, generally in inert solvents, in the presence of an azodicarboxylate, optionally in the presence of an additional reagent, preferably in a temperature range from 0 ° C. to room temperature normal pressure.

Additional reagents are customary additional reagents for Mitsunobu reaction conditions, such as triphenylphosphine, diphenyl- (2-pyridyl) -phosphine or (4-dimethylaminophenyl) -diphenylphosphine, triphenylphosphine is preferred.

Azodicarboxylates are, for example, diethyl azodicarboxylate, dimethyl azodicarboxylate, diisopropyl azodicarboxylate or di-tert-butyl azodicarboxylate, diethylazodicarboxylate is preferred. Inert solvents are, for example, halogenated hydrocarbons such as chloroform, ethers such as dioxane, tetrahydrofuran or 1,2-dimethoxyethane, hydrocarbons such as benzene, xylene or toluene, nitroaromatics such as nitrobenzene, carboxylic acid amides such as dimethylformamide or dimethylacetamide, alkyl sulfoxides such as dimethyl sulfoxide, aliphatic nitrile nitrate, such as acetonitrile nitrile , Esters such as ethyl benzoate or other solvents such as N-methylpyrrolidone, preference is given to tetrahydrofuran.

The compounds of the formula (III), (V), (VI), (IX), (XI) and (XII) are known or can be prepared by known processes.

The compounds of the formula (II), (IV) and (VII) can, if appropriate, be obtained from the reaction mixtures by chromatography.

The compounds of the formula (VIII) are known (R ¹⁰ = CH ₃ : Cadet, J; Teoule, R.

Bull. Soc. Chim. 1974, 1565-1570) or can be prepared, for example, according to synthesis scheme 2 (for 1st stage see J Am. Chem. Soc. 1997, 119, 2795-2803).

Synthesis scheme 2:

(VI)

The process according to the invention can be illustrated by the following synthesis schemes. Synthesis scheme 3:

Synthesis scheme 4:

The invention further relates to processes for the preparation of the compounds of

formula

in which

R ^{2 represents} hydroxy or acetoxy,

R ⁶ and R ⁹ independently of one another represent hydrogen or chlorine,

characterized in that the compounds of formula (Id) are obtained by fermentation of microorganisms and the subsequent isolation of the compound from the culture broth.

The compounds of the formula (Id) are particularly preferably prepared by fermentation of Basidiomycetes, organisms of the genus Agaricus are particularly preferably used here.

For the breeding of Agaricus spp. all common culture methods can be used that lead to the production of sufficient biomass.

In general, Agaricus sp. and fermentation to the compounds regardless of the type of containers, fermenters and

Starting conditions. It is advantageous to put a vegetative inoculum in an aqueous

Culture medium by inoculating this medium with an aliquot of a soil or

To produce slant tube culture. Having a fresh, active, vegetative

Inoculum has been prepared in this way, this is aseptically into further shaking bottles or other suitable equipment for the fermentation of the

Microorganisms transferred. A vegetative inoculum is preferably used for large-volume fermentations. The vegetative inoculum is made by inoculating a small volume of the culture medium with mycelium fragments. The vegetative inoculum is then transferred to a fermentation vessel in which, after a suitable incubation period, the compounds according to the invention are produced in an optimal yield. The medium in which the vegetative Inoculum produced can be identical or different to that medium which is used for the preparation of the compounds according to the invention, as long as it ensures sufficient microorganism growth.

The mushrooms are fermented successfully in NMG media (e.g. glucose:

0.4%, malt extract: 1.0%, yeast extract: 0.4%) or malt extract media (malt extract 1.0% to 4.0%). Also other culture media such as medium Q6 / 2, ZM / 2, MGP medium (all see example 1), as well as oatmeal medium, potato dextrose nutrient solution, CMG nutrient solution and commeal medium (the latter culture media see ATCC Media Handbook , American Type Culture Collection, Rockville, Md., USA

1984) are suitable.

Typically, a strain from the genus Agaricus is fermented in an aqueous nutrient medium under aerobic conditions, e.g. in a nutrient medium containing a source of coblene and possibly a proteinaceous material. Other carbon sources include glucose, brown sugar, sucrose, glycerin, starch, corn starch, lactose, dextrin, molasses, etc. Preferred nitrogen sources include cottonseed flour, yeast, autolysed baker's yeast, solid milk components, soybean meal, corn flour, pancreatic or papainically digested casein breakdown products, solid distillation components , Animal broths

Peptone, pieces of meat and bones etc. Trace elements, such as zinc, magnesium, manganese, cobalt, iron etc. do not have to be added to the fermentation medium as long as tap water and non-purified components are used as medium components.

The preparation of the compounds of the formula (Id) can be induced at any temperature which ensures sufficient growth of the microorganisms. The temperature is preferably between 21 ° C. and 32 ° C., particularly preferably approximately 24 ° C. In an aerobic submerged fermentation process, sterile air is usually passed through the culture medium. For efficient growth of the microorganisms, the volume of air used is in the range from approximately 0.25 to approximately 0.5 volume of air per volume of culture medium per minute (wm). An optimal ratio in a 10 L boiler is approximately 0.2 wm with movement generated by a conventional propeller rotating at approximately 200-500 rpm, preferably 100 rpm. The addition of a small amount, such as 1 ml / l, of an anti-foaming agent, such as silicone, to the fermentation medium is necessary if foaming is a problem.

A further possibility for the preparation of the compounds of formula (Id) consists in the cultivation of microorganisms of the genus Agaricus on solid substrates, by adding gelatinizing materials such as agar-agar nutrient solutions or in liquid media in standing culture, for which purpose all common substrates including the liquid media specified above can be used.

In general, optimal production of the compounds of formula (Id) is obtained within 8 to 25 days, preferably in about 11 days.

The fermentation broth usually becomes acidic during the fermentation (pH

4.5 to pH 2.5). The final pH is partly dependent on the buffer that may be used and partly on the initial pH of the culture medium. The pH is preferably adjusted to approximately 6.0 to 7.0 before sterilization, particularly preferably to pH 6.3.

Preferred fermentation conditions and media are described in the examples.

The compounds according to the invention are mainly present in the mycelium of the fermented microorganisms, but they can also occur, generally in smaller amounts, in the culture supernatant of the microorganisms. The culture broth can be easily obtained by filtering through a filter press or a nutsche.

The production of the compounds according to the invention can be followed during the fermentation with the aid of analytical HPLC (UV or mass spectrometric detection), the compounds according to the invention being detectable directly in the crude extract. For this purpose, extracts from freeze-dried culture broths as well as extracts made with the help of liquid-liquid extraction from smaller amounts of mycelium or culture broth removed during the fermentation can be used.

The production of the compounds according to the invention can be increased by sterile addition of potential biogenetic precursors such as glycerol and 3,5-dichloro-4-hydroxybenzoic acid to the fermentation medium.

The compounds according to the invention are suitable for use as medicaments for the treatment and / or prophylaxis of diseases in humans and / or animals.

The compounds according to the invention show an unforeseeable, valuable spectrum of pharmacological activity.

They are characterized as neurolysin inhibitors.

Because of their pharmacological properties, the compounds according to the invention can be used alone or in combination with other medicaments for the prophylaxis and treatment of acute and / or chronic pain (for a classification see "Classification of Chronic Pain,

Descriptions of Chronic Pain Syndromes and Definitions of Pain Terms ", 2nd ed., Meskey and Begduk, eds .; IASP-Press, Seattle, 1994), in particular for the treatment of cancer-induced pain and chronic neuropathic Pain such as diabetic neuropathy, post-herpetic neuralgia, peripheral nerve damage, central pain (e.g. as a result of cerebral ischemia) and trigeminal neuralgia, and other chronic pain such as lumbago, low back pain, inflammatory or rheumatic Pain. These are also suitable

Substances also for the therapy of primary acute pain of any genesis and the secondary pain states resulting therefrom, as well as for the therapy of chronic, formerly acute pain states.

The invention also relates to eight new strains (WP 4080, WP 4105,

WP 0058, WP 4385, WP 4140, WP 0059, FU 41954 and WP 4121) of the aforementioned fungi of the genera Agaricus, Hypholoma and Stropharia.

This invention also includes the use of any mutants capable of producing agaricoglycerides, including natural mutants and artificial mutants, which can be obtained in a manner known per se from the said microorganisms by conventional means, e.g. by radiation with X-rays, ultraviolet radiation, treatment with N-methyl-N'-nitro-N-nitrosoguanidine, 2-aminopurine and the like or by methods of genetic engineering.

The in vitro activity of the compounds according to the invention can be shown in the following assays:

1. Neurolysin Inhibition Assav

To determine their in vitro effect on neurolysin (endopeptidase EC 3.4.24.16), the test substances are dissolved in 100% DMSO and serially diluted. Typically, dilution series from 500 μM to 3.9 μM are produced (resulting final concentrations in the test: 10 μM to 0.078 μM). 2 μL of the diluted substance solutions are placed in the wells of microtiter plates (OptiPlate-96; Packard) submitted. Then 50 μL of a diluted neurolysin solution are added. The dilution of the neurolysin is chosen so that less than 70% of the substrate is converted during the later incubation (typical neurolysin final concentration fc = 1.5 nM; dilution buffer: 50 mM Tris / HCl pH 7.5, 0.0025% Brij-35). The peptide substrate, DABCYL-Glu-Arg-Nle-Phe-Leu-Ser-Phe-Pro-EDANS; is diluted with assay buffer (50 mM Tris / HCl pH 7.5) to a concentration of 10 μM (final substrate concentration fc = 5 μM). The enzyme reaction is finally started by adding 50 μL of diluted peptide substrate. The test batches are incubated for 90 min at room temperature and then measured in a FluorStar fluorimeter (BMG LabTechnologies) (λex = 320 nm, λem = 520 nm). IC ₅₀ values are determined on the basis of the graphical plot of the substance concentration against the percentage inhibition.

Representative in vitro activity data for the compounds according to the invention are in

Table A reproduced:

Table A

The suitability of the compounds according to the invention for the treatment of pain conditions can be shown in the following animal models: 2. Axotomy of the Tibial Nerve in the Rat (Chronic Pain Model)

Under pentobarbital anesthesia, the trifurcation of a sciatic nerve is dissected and the branch of the tibial nerve axotomized. Control animals are given a sham operation. After the operation, the axotomized animals develop chronic mechanical AUodynia and thermal hyperalgesia.

Mechanical AUodynia is tested using a pressure transducer in comparison to sham operated animals (electronic from Frey Anesthesiometer, IITC Inc-Life Science Instruments, Woodland Hills, CA, USA).

Thermal hyperalgesia can be determined by measuring the latency time within which a rat removes a paw from the area of a radiant heat source (Plantartest, Ugo Basile (Comerio, Italy)).

The substance is administered at different points before the pain test via different application routes (i.V., i.p., p.o., i.t, i.c.v., s.c, transdermal).

For the quantitative evaluation of the analgesic effect of a test substance, the difference between the von Frey pressures and the latency times in the plantart test is first determined between sham-operated and vehicle-treated axotomized rats. This difference represents the maximum achievable effect of the tibial axotomy. The effect of an analgesic is shown as a percentage change in pain parameters related to the maximum effect.

The new active ingredients can be converted in a known manner into the customary formulations, such as tablets, dragées, pills, granules, aerosols, syrups, emulsions, suspensions and solutions, using inert, non-toxic, pharmaceutically suitable excipients or solvents. The therapeutically active compound should in each case be present in a concentration of about 0.5 to 90% by weight of the Total mixture must be present, ie in amounts sufficient to achieve the dosage range indicated.

The formulations are, for example, by stretching the active ingredients with solvents and or carriers, optionally using

Emulsifiers and / or dispersants, e.g. in the case of the use of water as a diluent, organic solvents can optionally be used as auxiliary solvents.

The application is carried out in the usual way, preferably orally, transdermally or parenterally, in particular perlingually or intravenously. However, it can also be done by inhalation through the mouth or nose, for example with the aid of a spray, or topically via the skin.

In general, it has proven to be advantageous for parenteral administration

To administer amounts of about 0.01 to 30, preferably about 0.1 to 20 mg / kg body weight for effective results. In the case of oral administration, the amount is approximately 0.1 to 30 mg / kg, preferably approximately 0.5 to 10 mg / kg body weight.

Nevertheless, it may be necessary to deviate from the amounts mentioned, depending on the body weight or the type of application route, on the individual behavior towards the medication, the type of its formulation and the time or interval at which the administration takes place , In some cases it may be sufficient to make do with less than the aforementioned minimum quantity, while in other cases the above upper limit must be exceeded. In the case of application of larger quantities, it may be advisable to distribute them in several individual doses over the day.

Unless otherwise stated, all quantities relate to percentages by weight. Solvent ratios, dilution ratios and concentration details of liquid / liquid solutions each relate to the volume. The "W / v" means "weight / volume". For example, "10% w / v" means: 100 ml of solution or suspension contain 10 g of substance.

Characteristics of the producer strains

The strains WP 4080 and WP 4105 were made using medium 2 from in Wuppertal in summer 1999, each on meadows by Dr. M. Stadler found and identified fruiting bodies of Basidiomycetes isolated. While the WP 4080 strain was isolated from the spore imprint of a Basidiomycete fruiting body, the WP4105 strain was successfully extracted from the sterile context of the

To take hat of another Basidiomycete fruiting body in culture. The fruiting bodies showed a similar morphology and, according to the available knowledge, belong to the same species. They were divided into stems and hats and initially had pink-colored, later black-colored lamellas, on which the basidia with the basidiospores were formed. The spore dust was black. In boys

A velum was observed in the fruiting bodies, and the adult fruiting bodies accordingly had a simple ring on the stem, which originated from the remnants of the velum. The stem was 1-2 cm in diameter and up to 7 cm long, and the initially hemispherical hat, spread out in older specimens, reached a diameter of up to 8 cm. A characteristic yellowing was observed on the outside of the entire fruiting body under pressure. These characteristics made it possible to assign the mushrooms to the genus Agaricus L. ex Fr. According to the identification key from Moser, M. (The Röhrlinge und Blätterpilze (ed. W. Garns) Kleine Kryptogamenflora Band 2b / 2, Fischer Verlag Stuttgart (1983)) the mushroom belongs to the Flavescentes section of the genus Agaricus. After the key on p. 226f. both materials of the species Agaricus macrosporus (Moeller & J. Schaeffer) Pilät were assigned to the aforementioned literature.

For the morphological characterization, the fungal cultures were grown on YMG agar (medium la). After a week, the mycelium reached about 7 cm in diameter on an agar plate of 9 cm. The substrate discolored after about 10 days brownish, whereas the mycelium remained white-felted. Buckles were observed on the septa of the hyphae.

Both strains are deposited under the accession number DSM 14593 for the tribe WP 4080 and under the accession number DSM 14594 for the tribe

WP 4105 (the filing date is 31.10.2001 in both cases) under the Budapest Treaty with the DSMZ, German Collection of Microorganisms and Cell Cultures GmbH, Mascheroder Weg lb, D-38124 Braunschweig, Germany.

Following General Procedure A and then applying the

General Procedure C can also be found in other strains of Basidiomycetes producing agaricoglycerides. These organisms are listed in detail below: The compounds according to the invention were not isolated from these strains, but with the aid of their HPLC characteristics (UV spectrum, ESI spectra and retention times in two different HPLC-

Gradient systems), as well as the biological test of the extracts for neurolysin inhibition.

The strain MUCL 35030 was created in 1992 by Dr. C. Decock in Warcoing (Belgium) found in a garden and called Agaricus arvensis Schaeff.:Fr. identified fruiting bodies isolated and supplied to Bayer AG by MUCL (BCCMTM / MUCL, Mycotheque de l'Universite catholique de Louvain, Place Croix du Sud 3, B-1348 Louvain-la-Neuve, Belgium). The strain was again deposited with the DSMZ under the Budapest contract under the number DSM 14892. The filing date is March 8, 2002.

The WP 0058 strain was developed by Dr. H. Müller (Bayer AG) in September 1994 isolated from the spores of fruiting bodies of Agaricus bisporus (Lange) Imbach found in Wuppertal in September 1991. The strain was deposited with the DSMZ under the number DSM 14900 under the Budapest contract.

The filing date is March 18, 2002. The fruiting bodies of the herbarium STMA 98023 were developed on May 9, 1998 by Dr. M. Stadler in Hamburg near the Volksparkstadion found on a meadow and as Agaricus bitorquis (Quel.) Sacc. identified. The culture (strain WP 4385) was created from the context of the hat. The trunk was numbered

DSM 14895 deposited with the DSMZ under the Budapest contract. The filing date is March 8, 2002.

The strain MUCL 28516 was created in 1984 by Prof. Dr. G. L. Hennebert in Kapellen (Belgium) and found as Agaricus bitorquis (Quel.) Sacc. identified fruiting bodies isolated and supplied to Bayer AG by MUCL (BCCMTM / MUCL, Mycotheque de l'Universite catholique de Louvain, Place Croix du Sud 3, B-1348 Louvain-la-Neuve, Belgium). The strain was again deposited with the DSMZ under the Budapest contract under the number DSM 14893. The filing date is March 8, 2002.

The fruiting bodies of the STMA 97106 herbarium were obtained on September 9, 1997 in Wuppertal (Pahlkestrasse) from Dr. M. Stadler found on the edge of a paved walkway and as Agaricus bitorquis (Quel.) Sacc. identified. The culture (strain WP 4140) was created from the context of the hat. The strain was deposited with the DSMZ under number DSM 14894. The filing date is March 8, 2002.

The strain MUCL 29004 was created in 1986 by Prof. Dr. GL Hennebert in Louvain-la-Neuve (Belgium) found on a meadow and called Agaricus campestris var. Campestris L.:Fr. identified fruiting bodies isolated and supplied to Bayer AG by MUCL (BCCMTM / MUCL, Mycotheque de l'Universite catholique de Louvain, Place Croix du Sud 3, B-1348 Louvain-la-Neuve, Belgium). The strain was again deposited with the DSMZ under the Budapest contract under the number DSM 14896. The filing date is March 8, 2002. The strain MUCL 28242 was created in 1993 by Prof. Dr. GL Hennebert in the Bois de Boulaide (Luxembourg) and fruit bodies identified as Agaricus macrosporus (Moeller & J. Schaeffer) Pilät isolated and isolated from the MUCL (BCCMTM / MUCL, Mycotheque de l'Universite catholique de Louvain, Place Croix du Sud 3, B -1348 Louvain-la-Neuve, Belgium) to Bayer AG. The strain was again with the DSMZ under the Budapest contract under the number DSM

14897 deposited. The filing date is March 8, 2002.

The strain CBS 585.76 was developed in September 1976 in Empolder near Embrugge (Netherlands) by Prof. Dr. W. Garns found and as Agaricus macrosporus

(Moeller & J. Schaeffer) Pilät identified fruiting bodies isolated and delivered to Bayer AG by CBS (Uppsalalaan 8, P.O.Box 85167, 3508 AD Utrecht, Netherlands). The strain was again deposited with the DSMZ under the Budapest contract under the number DSM 14921. The filing date is April 8, 2002.

The WP 0059 strain was developed by Dr. H. Müller (Bayer AG) in September 1994 from the spores of fruiting bodies of Agaricus xanthoderma Genev found in Wuppertal in September 1991. isolated. The strain was deposited with the DSMZ under the number DSM 14901 under the Budapest contract. The filing date is March 18, 2002.

The fruiting bodies of the herbarium STMA 00034 were grown on September 9, 1997 in Wuppertal by Dr. M. Stadler in the Finsterbruimertal district (Schopp, Rhineland-Palatinate) found on wood from Pseudotsuga and identified as Hypholoma marginatum (Pers.:Fr.) J. Schrot. The culture (strain FU41954) was created from the context of the hat. The strain was numbered DSM

14898 deposited with the DSMZ under the Budapest contract. The filing date is March 8, 2002.

The WP 0360 strain was obtained from Prof. R. Seeger on September 25, 1992 from fruiting bodies of a Hypholoma sp. Found near Coburg. isolated and to the Bayer AG delivered. The strain was deposited with the DSMZ under the number DSM 14902 under the Budapest contract. The filing date is March 18, 2002.

The strain FU 40473 was developed by Dr. H. Dörfelt isolated on October 3, 1999 from fruiting bodies of Psathyrella prona (Fr.) Gillet found on a cattle pasture in Neuengönna (Thuringia) and delivered to Bayer AG. The strain was deposited with the DSMZ under the number DSM 14903 under the Budapest contract. The filing date is March 18, 2002.

The strain DSM 11373 was supplied by the DSMZ as Stropharia rugosoannulata Farlow ex Murrill to Bayer AG and again deposited with the DSMZ under the Budapest contract under the number DSM 14904. The filing date is March 18, 2002.

The fruiting bodies of the herbarium STMA 97126 were harvested on September 22, 1997 near Johanniskreuz / Pfalz near Burgalbspring by Dr. M. Stadler found and as Stropharia squamosa (Pers.:Fr.) Quel. identified. The culture (strain WP 4121) was created from the basidiospores. The strain was deposited with the DSMZ under the number DSM 14899 under the Budapest contract.

The filing date is March 8, 2002.

In the course of these investigations it was surprisingly found that the production of agaricoglycerides not only with the help of fungi of the species Agaricus macrosporus, but also with the help of fungi of other types of

Genus Agaricus, as well as using mushrooms of the genera Psathyrella, Hypholoma and Stropharia succeed. According to P.M. Kirk et al. (eds.): Dictionary of the Fungi, Ninth Edition, CABI Publishing, Wallingford U.K. (2001) different families of the Basidiomycetes, namely the Agaricaceae (Agaricus), Strophariaceae (Hypholoma and Stropharia), and the Coprinaceae

(Psathyrella). These studies show that the production of Compounds according to the invention are surprisingly widespread in Basidiomycetes and indicate that agaricoglycerides can also be found in other families, genera and species.

Abbreviations:

ACN acetonitrile aq. Aqueous abs. Absolutely

Amu atomic mass unit

Bn benzyl

Boc tert-butoxycarbonyl

DABCYL 4- (4-dimethylaminophenylazo) benzoyl

DMSO dimethyl sulfoxide

EDANS 5 - [(2-aminoethyl) amino] naphthalene-1 sulfonyl d. Th. Of theory eq. equivalent to

ESI electrospray ionization (for MS) sat. Saturated

H hour

HPLC high performance liquid chromatography

LC-MS liquid chromatography-coupled mass spectroscopy

Lit. reference min minutes

MS mass spectroscopy

MeOH methanol

NMR nuclear magnetic resonance proc. percent

RT room temperature, 20 ° C

Rt retention time (with HPLC)

TFA trifluoroacetic acid

THF tetrahydrofuran

Tris tris (hydroxymethyl) aminomethane cf. compare fermentation media

Medium 1 (YMG; yeast-malt-glucose medium): glucose: 0.4%, yeast extract 0.4%, malt extract 1.0%, made up with 1 L tap water. Before sterilization at 121 ° C for 20 min, the pH was adjusted to 6.3 with 0.1 M HC1.

Medium la (YMG Agar): 1.5% Difco Bacto Agar was added to Medium 1 as described above before autoclaving. After cooling, slanted tubes or agar plates were poured from it.

Medium 2 (Mycorrhiza Medium): malt extract 0.8%, glucose 0.7%, asparagine 0.05%, casein hydrolyzate 0.1%, KH ₂ PO4 0.05%, MgSO ₄ 0.05%, yeast extract 0.1%, Difco Bacto Agar 1.5% ad 1 L deionized water, pH Set 5.05 with 1 N hydrochloric acid before autoclaving for 30 min at 121 ° C and 1 bar

Overpressure, when cooling to 55 ° C addition of the antibiotics streptomycin sulfate and penicillin G in final concentrations of 125 mg / L each). After cooling, slanted tubes or agar plates were poured from it, which served to isolate the cultures.

Medium 3 (ZM / 2): molasses 1%, oatmeal 1%, glucose 0.3%, sucrose 0.8%, mannitol 0.8%, edamin 0.1%, (NH) ₂ SO ₄ 0.1%, CaCO ₃ 3%, tap water ad 1 L , pH 7.2 before autoclaving for 30 min at 121 ° C with 0.1 N sodium hydroxide solution. Medium 4 (MGP medium): glucose 1%, maltose 2%, soy peptone 0.2%, yeast extract 0.1%, KH ₂ PO ₄ 0.1%, MgSO ₄ x 7 H ₂ O 0.05%, FeCl ₃ (10 mM), 1 ml , ZnSO (1.78 g / 1) 1 ml, CaCl ₂ (0.1 mM) 1 ml).

Medium 5 (Q6 / 2): glucose 2.5%, glycerin 1%, cottonseed flour 0.5%, CaCO ₃

0.05%, ad 1 L tap water, pH not adjusted, autoclaved for 30 min at 121 ° C and 1 bar overpressure.

The above information applies to media in shake cultures and agar plates. The fermenters were generally sterilized with steam and with the addition of small amounts of defoamer, as described below in the corresponding examples.

General driving A: Production of agaricoglycerides with the strains Asaήcus sp. WP 4080 and WP4105 in the piston

The fermentation was carried out using 1 L Erlenmeyer flasks, each containing 300 ml of medium 1, which was inoculated with 10 pieces of agar from a well-covered plate of medium la, each about 1 cm in diameter. Either Agaricus sp. WP 4080 or Agaricus sp. WP

4105 used. The fermentation was carried out at an incubation temperature of 23 ° C. on a rotary shaker at 140 rpm for 192 hours. The mycelium was separated from the rest of the culture broth by centrifugation (15 min at 1000 x g) and then extracted as described in general procedure C.

Analogous fermentations in media 2 - 5 also led to the formation of the agaricoglyceride after a fermentation period of 192 h. This was determined with the help of HPLC analyzes to quantify the agaricoglyceride (see Table 2). With media M4 and M5, the fermentation time can be extended cause an increase in the agaricoglyceride content up to 300 h, so that these media have been selected for the fermentations on a 30 L scale.

The strain Agaricus sp. WP 4080 is characterized by better production rates and was therefore selected for the production in the stirred tank, which is described in the following

General Procedure B is described.

General driving B: Production of agaricoglycerides with the strain Agaricus sp. WP 4080 by fermentation in a stirred tank

The strain Agaricus sp. WP 4080 in medium 1 (see under fermentation media), as described under general procedure A, is grown.

Approx. 10 pieces of a mycelium culture on solid medium were used as inoculum for

Prepare 200 ml of medium 1 in a 1000 ml Erlenmeyer flask and incubate on a rotary shaker for 173 hours at 140 revolutions per minute (rpm). These cultures served as an inoculum for production cultures A and B.

Production culture A (30 L scale)

The fermentation was carried out either in medium 4 or medium 5. To prepare the production culture, 1 x 200 ml (for main culture in medium 4) or 3 x 200 ml (for main culture in medium 5) of a preculture was used as an inoculum for 30 L medium 4 or medium 5. Before inoculating the medium

1 ml defoamer SAG 5693 (Union Carbide, USA) per L, and sterilized for 30 min at 1.1 bar with steam. This production culture was incubated in a 40 L Biostat E (Braun Bioengeneering, Melsungen) for 331 hours (medium 4) and 233 hours (medium 5) in each case at 24 ° C., a stirring speed of 150 rpm and with an aeration rate of 0.3 wm , Production culture B (200 L scale)

To prepare the production culture on a 200 L scale, 2 × 350 ml of a preculture were first used as an inoculum for 20 L medium 5. Before the inoculation, 1 ml of defoamer SAG 5693 (Union Carbide, USA) per L was added to the medium, and sterilized with steam for 60 min at 1.1 bar. This preculture II was inhibited for 180 hours at 24 ° C. at a stirring speed of 150 rpm and with an aeration rate of 0.2 wm in a 30 L bioengineering ring fermenter (blade stirrer). The 20 L preculture II was used to inoculate 200 L sterile medium 5. Before inoculation, 1 ml of defoamer was added to the medium

SAG 5693 (Union Carbide, USA) added per L, and sterilized with steam for 60 min at 1.1 bar. This production culture was incubated for 115 hours at 24 ° C., aeration of 0.2 wm and a stirrer speed of 100 rpm.

Accompanying fermentation analysis

Daily samples of 50 ml were used to monitor the production process in shake cultures and fermenters, which were taken sterile and extracted as described and analyzed using analytical HPLC.

Extraction and sample preparation: The mycelium from 50 ml culture broth was filtered off and extracted with 50 ml acetone in an ultrasonic bath for 30 min. The acetone was concentrated on a rotary evaporator at 250 mbar and the aqueous residue was diluted to 50 ml of water. This sample was extracted with 50 ml of ethyl acetate. 5 ml of the organic phase was removed over anhydrous

Na ₂ SO filtered and first concentrated to dryness using a vacuum centrifuge, then taken up in 1 ml of methanol and filtered sterile. Aliquots of 1-5 μl of the extracts produced in this way were examined on analytical HPLC. HPLC parameters (fermentation analysis): Two different analytical HPLC methods with UN / visual (HPLC-UV / Vis) and with mass spectrometric detection (HPLC-MS) were used to characterize the agaricoglycerides.

HPLC-UN / Nis analyzes were carried out using a Hewlett Packard Series 1100 analytical HPLC system (HP, Waldbronn, Germany), consisting of a G 1312A binary pump system, a G 1315A diode array detector, a G 1316A column temperature control system, a G 1322A degassing system and one

G 1313A auto-injector performed. 0.01% H ₃ P0 ₄ : acetonitrile (ACΝ) was used as the mobile phase at a flow of 1 ml / min, while a Merck (Darmstadt, Germany) Lichrospher RP 18 column (125 x 4 mm, particle size 5 μm) was used as the stationary phase served. The samples were run on a continuous linear gradient (0% ACΝ to 100% ACΝ in 10 min), followed by isocratic

Elution (5 min at 100% ACΝ) separated. HPLC-UN chromatograms were recorded at 210 nm with a reference wavelength of 550 nm and a bandwidth of 80 nm. The diode array detection in the range of 210-600 nm provided the HPLC / UV / Vis spectra. The data was saved and evaluated using the HP ChemStation software.

HPLC-MS was carried out on an HP 1100 HPLC system, coupled to a Micromass-LCT mass spectrometer (Micromass, Manchester, Great Britain) in ESI + mode and in ESIneg mode. Mass spectra were recorded in the range between 200 and 1200. The HP100 system used operated with the following parameters: Stationary phase: Waters Symmetry C18, 3.5 around 2.1x50mm column, mobile phase: gradient water (A), acetonitrile (B) + 0.1% formic acid (0-1 min 100% A; 1-5 min linear gradient to 10% A / 90% B; 5-6 min 10% A / 90% B, 6-6.10 min 90% B - 100% B). The results were saved and evaluated using the integrated Micromass OpenLynx Browser Version software

3.5. The qualitative detection was carried out by comparing the characteristic HPLC-UV and HPLC-MS data (retention times, UV and ESI-LC-MS spectra). The content determination for the agaricoglyceride A and the detection of the other compounds was carried out using external and internal standards of methanolic solutions of the agaricoglycerides. The retention times of the compounds are shown in Table 1 and the amounts of agaricoglyceride A formed during the fermentation in Table 2. Since the secondary components (examples 2 and 3) were difficult to detect from the crude extracts, they were not quantified.

Table 1: Retention times (R _t ) of the agaricoglycerides on the analytical HPLC.

Table 2: Content of agaricoglyceride A in the mycelial extracts of WP 4080 from shake cultures in different media and with different fermentation times

General driving C: Qualitative detection of agaricoglycerides in producer strains - HPLC screening

To check Agaricus spp. and other basidiomycete cultures based on agaricoglyceride production, either freeze-dried materials or fresh cultures were extracted and analyzed using analytical HPLC.

a) Qualitative detection from freeze-dried mushroom cultures

Cultures of Agaricus spp. and other mushrooms were fermented in medium Ml until the free glucose was consumed and then freeze-dried. Aliquots of 50 mg of the resulting lyophilisates were mixed with 5 ml dichloromethane methanol 1: 1 and extracted for 30 min in an ultrasonic bath at 40 ° C. The lyophilisate was filtered off and the extracts were evaporated to dryness using a vacuum centrifuge. The oily residues were then distributed in 5 ml of water / ethyl acetate 1: 1 for 15 min in an ultrasonic bath. An aliquot of 1 ml of the organic phase was removed, filtered over anhydrous sodium sulfate and concentrated to dryness. The residue was then taken up in 200 .mu.l of methanol, filtered and 10 ml thereof was analyzed for the presence of agaricoglyceride on analytical HPLC. The qualitative detection of agaricoglycerides was carried out by comparing the characteristic HPLC-UV and HPLC-MS

Data (retention times, UV and ESI-MS spectra).

b Direct qualitative detection from liquid cultures

Cultures of Agaricus spp. and other mushrooms were fermented as described above until the free glucose in the medium was consumed. Then 10 ml of the cultures were removed and extracted with 10 ml of ethyl acetate for 30 min in an ultrasonic bath. 5 ml of the organic phase were removed, filtered over anhydrous sodium sulfate and first concentrated to dryness with the aid of a vacuum centrifuge, then taken up in 1 ml of methanol, filtered and analyzed for the presence of agaricoglyceride on analytical HPLC. The qualitative detection was carried out by comparing the characteristic HPLC-UV and HPLC-MS data (retention times, UV and ESI-MS spectra).

Analytical methods: HPLC methods

Method A: Instrument: HP 1100 with DAD detection; Column: Kromasil RP-18, 60 mm x 2 mm,

3.5 μm; Eluent: A = 5 ml perchloric acid / 1 water, B = acetonitrile; Gradient: 0 min 2% B, 0.5 min 2% B, 4.5 min 90% B, 15 min 90% B; Flow: 0.75 ml / min, temp .: 30 ° C, detection UV 210 nm.

Method B:

Instrument: HP 1100 with DAD detection; Column: Kromasil RP-18, 60mm x 2mm, 3.5 μm; Eluent: A = 5 ml perchloric acid / 1 water, B = acetonitrile; Gradient: 0 min 2% B, 0.5 min 2% B, 4.5 min 90% B, 6.5 min 90% B; Flow: 0.75 ml / min, temp .: 30 ° C, detection UV 210 nm.

Method C:

Instrument: HP 1100 with DAD detection; Column: Kromasil RP-18, 60mm x 2mm, 3.5 μm; Eluent: A = 5 ml perchloric acid / 1 water, B = acetonitrile; Gradient: 0 min 2% B, 0.5 min 2% B, 4.5 min 90% B, 9 min 90% B; Flow: 0.75 ml / min, temp .: 30 ° C, detection UV 210 nm.

LC / MS methods

Method 1: HPLC-MS is connected to an HP 1100 HPLC system, coupled to a micromass

LCT mass spectrometer (Micromass, Manchester, Great Britain) performed in ESI + and ESI mode. Mass spectra are recorded in the range between 100 and 1650 amu. The HPllOO system used operates with the following parameters: Stationary phase: Waters Symmetry C18, 3.5 around 2.1 x 50 mm column, mobile phase: eluent: water + 0.1% formic acid (A), acetonitrile + 0.1%

Formic acid (B); Gradient: 0-1 min 100% A; 1-5.5 min linear gradient to 5% A / 95% B; 5.5-8 min 5% A / 95% B, 8-8.1 min 0% A - 100% B, 8.1-10 min 0% A- 100% B; Flow: 0.5 ml / min.

Method 2: Instrument: Micromass ZMD, HP1100; Column: Nucleosil, 50 mm x 2.1 mm, 5 µm;

Eluent A: acetonitrile + 0.05% formic acid, eluent B: water + 0.05% formic acid; Gradient: 0.0 min 5% A - »12.0 min 100%, A ^■ » 15.0 min 100% A; Oven: 35 ° C, flow: 0.5 ml / min, UV detection: 210-500 nm.

starting compounds

Example 1A

2 - {[4- (benzyloxy) -3,5-dichlorobenzoyl] oxy} -l - ({[4- (enzyloxy) -3,5-dichlorobenzoyl] - oxy} methyl) ethyl-4- (benzyloxy) - 3,5-dichlorobenzoate

150 mg (1.63 mmol) glycerol and 0.90 ml (8.14 mmol) triethylamine are dissolved in 50 ml abs. Submitted dichloromethane. At 0 ° C 2.57 g (8.14 mmol) 4-

Benzyloxy-3,5-dichlorobenzoyl chloride added. The suspension is warmed to RT in an ice bath and stirred for 5 days. After the suspension has been filtered through silica gel 60 (mobile phase: ethyl acetate), the filtrate is concentrated and the residue is stirred with cyclohexane. The solid is filtered off, boiled in methanol and the remaining solid is suctioned off again. After dissolving in dichloromethane, the solution is washed with sodium bicarbonate solution, dried and concentrated. The residue is purified on silica gel 60 (mobile phase: cyclohexane / ethyl acetate 100: 0, 100: 2, 100: 4, 100: 6, 100: 8, 100: 10). 210 mg (13.7% of theory) of product are obtained.

MS (MALDI): m / z = 933 (M + Li) ⁺ .

1H-NMR (400MHz, CDC13): δ = 8.00 (s, 2H), 7.96 (s, 4H), 7.57-7.51 (m, 5H), 7.44-7.35 (m, 10H), 5.79-5.72 (m, 1H ), 5.12, 5.11 (2s, 6H), 4.76 (dd, 2H), 4.63 (dd, 2H).

Example 2A

3 - {[4- (Benzyloxy) -3,5-dichlorobenzoyl] oxy} -2-hydroxypropyl-4- (benzyloxy) -3,5-di-chlorobenzoate

1 g (10.86 mmol) glycerin is mixed with 10 ml abs. Pyridine coevaporated and then in 30 ml abs. Pyridine dissolved. The solution is cooled to 0 ° C. under argon, 7.2 g (22.8 mmol) of 4- (benzyloxy) -3,5-dichlorobenzoic acid chloride (Example 12A) are added, the mixture is slowly warmed to room temperature and stirred at room temperature overnight. The reaction mixture is concentrated, coevaporated with toluene and purified by chromatography (silica gel 60, mobile phase: cyclohexane-ethyl acetate 10: 1, 5: 1; 2: 1, 1: 1). 3.5 g (50% of theory) of product are obtained.

HPLC (method C): R _t = 6.24 min; MS (DCI, NH ₃ ): mz = 666 (M + NH ₄ ) ⁺ Example 3A

2- [(3,5-dichlorobenzoyl) oxy] -1 - (hydroxymethyl) ethyl-3,5-dichlorobenzoate

1.50 g (2.46 mmol) of 2,3-bis [(3,5-dichlorobenzoyl) oxy] propyl-3,5-dichlorobenzoate (prepared analogously to Example 1A from glycerol and 3,5-dichlorobenzoic acid chloride) are dissolved in 30 ml of DMF , After adding 165.3 mg (2.95 mmol) of solid potassium hydroxide, the mixture is stirred at room temperature under TLC control. The solvent is then removed in vacuo and the residue is purified on silica gel 60 (mobile phase: cyclohexane / ethyl acetate 100: 0, 100: 5, 100: 10, 100: 15, 100: 20, 100: 25, 100: 30). 75 mg (6.3% of theory) of product are obtained.

Example 4A

4- (benzyloxy) -3-chloro-5-fluorobenzoyl chloride

4- (Benzyloxy) -3-chloro-5-fluorobenzoic acid chloride can be prepared analogously to 4- (benzyloxy) -3,5-dichlorobenzoic acid chloride (Example 12A) starting from 4- (hydroxy) -3-chloro-5-fluorobenzoic acid (cf. : J Org. Chem. 1952, 17, 1425-1430). Example 5A

3- (benzyloxy) -4-chloro-6-fluorobenzoyl chloride

3- (Benzyloxy) -4-chloro-6-fluorobenzoic acid chloride can be prepared analogously to 4- (benzyloxy) -3,5-dichlorobenzoic acid chloride (Example 12A) starting from 3- (hydroxy) -4-chloro-6-fluorobenzoic acid (cf. EP -A 108526).

Example 6A

4- (benzyloxy) -3-cyanobenzoyl chloride

4- (Benzyloxy) -3-cyanobenzoic acid chloride can be prepared analogously to 4- (benzyloxy) -3,5-dichlorobenzoic acid chloride (Example 12A) starting from 4- (hydroxy) -3-cyano-benzoic acid (cf. US Pat. No. 4,151,297) , Example 7A

4- (benzyloxy) -3-fluorben-zoesäurechlorid

4- (Benzyloxy) -3-fluorobenzoic acid chloride can be prepared analogously to 4- (benzyloxy) -3,5-dichlorobenzoic acid chloride (Example 12A) starting from 4- (hydroxy) -3-fluoro-benzoic acid (cf. Lit. J Org. Chem. 1952, 17, 1425-1430).

Example 8A

4- (benzyloxy) -5-chloro-2,3-difluorbenzoesäurechlorid

4- (Benzyloxy) -5-chloro-2,3-difluorobenzoic acid chloride can be analogous to 4- (benzyloxy) -

3,5-dichlorobenzoic acid chloride (Example 12A) starting from 4- (hydroxy) -5-chloro-2,3-difluorobenzoic acid, obtainable from l-chloro-2,3,4-trifluoro-5- (trifluoromethyl) benzene (cf. DE-A 3 736089) in analogy to 3-chloro-2,5,6-trifluoro-4-hydroxybenzoic acid, which is obtained by heating l-chloro-2,3,4,6-tetrafluoro-5- (trifluoro - Methyl) benzene can be obtained with sodium hydroxide solution (cf. DE-A 3 804 288). Example 9A

4- (hydroxy) -3,5-dichlorobenzoic acid methylester

250 ml of methanol are introduced, cooled to -10 ° C and mixed with 65 ml (891 mmol) of thionyl chloride for 15 min. 50 g (241 mmol) of 4- (hydroxy) -3,5-dichlorobenzoic acid are then added and the mixture is stirred at room temperature for 1 hour and at reflux for 90 min. The solvent is distilled off and the

Residue coevaporated with 100 ml of methanol. It is recrystallized from 75 ml of ethanol. 18.6 g (35% of theory) of product are obtained. Another 33.9 g of methyl 4- (hydroxy) -3,5-dichlorobenzoate can be obtained by concentrating the mother liquor.

Example 10A

4- (benzyloxy) -3,5-dichlorobenzoic acid methylester

24 g (94 mmol) of 3,5-dichloro-4-hydroxy-benzoic acid methyl ester are dissolved in 200 ml of dichloromethane, 15.7 ml (112 mmol) of triethylamine and 12.3 ml (103 mmol) of benzyl bromide are added and the mixture is reacted for 3 days at room temperature. After adding water, the mixture is extracted with dichloromethane, dried and concentrated. 30 g of product are obtained, which is reacted further without purification.

Example 11 A

4- (benzyloxy) -3,5-dichlorobenzoic acid

36.3 g (116.6 mmol) of methyl 4- (benzyloxy) -3,5-dichlorobenzoate are dissolved in 150 ml of methanol, 150 ml of water and 7 g (175 mmol) of sodium hydroxide are added and the mixture is stirred at 50 ° C. overnight. The precipitate obtained after acidification to pH = 2 with 3 M hydrochloric acid is filtered off with suction, washed with water and dissolved in ethyl acetate. After shaking the organic phase with water and drying over magnesium sulfate, the mixture is concentrated. 27.36 g of product are obtained, which was reacted further without purification.

Example 12A

4- (benzyloxy) -3,5-dichlorbenzoesäurechlorid

19.2 g (64.6 mmol) of 4- (benzyloxy) -3,5-dichlorobenzoic acid are placed in 370 ml of toluene, mixed with 7 ml (96.9 mmol) of thionyl chloride at room temperature and stirred at 120 ° C. overnight. The reaction mixture is concentrated and coevaporated with toluene. 18.2 g (89% of theory) of product are obtained. Preparation Examples

example 1

2,3-bis [(3,5-dichloro-4-hydroxybenzoyl) oxy] propyl-3,5-dichloro-4-hydroxybenzoate (agaricoglyceride A)

Procedure 1:

15.0 mg (16.1 μmol) of Example 1A are dissolved in a mixture of 2 ml of methanol and 2 ml of tetrahydrofuran. After adding 5.0 mg of Lindlar catalyst, the mixture is hydrogenated for one hour at room temperature and RT. The suspension is filtered through Celite and washed with methanol. The filtrate is concentrated and the residue is purified on silica gel 60 (mobile phase: dichloromethane-methanol 100: 0, 100: 1, 100: 2). 2.8 mg (25.0% of theory) of product are obtained.

1H-NMR (400MHz, DMSO-d ₆ ): δ = 7.85 (s, 2H), 7.81 (s, 4H), 5.68-5.61 (m, 1H), 4.70 (dd, 2H), 4.60 (dd, 2H) , Method 2: Presentation from the mycelium of a shake flask fermentation (YMG medium)

The mycelium from 10 YMG shake flasks was separated from the supernatant by centrifugation and combined. The moist mycelium was then twice with each

1 L acetone extracted in an ultrasonic bath for 30 min each and filtered off. The filtered acetone-water mixture was evaporated in vacuo at about 40 ° C and 250 mbar. The resulting aqueous residue was diluted to 300 ml with tap water and then extracted three times with 300 ml of ethyl acetate each time. The organic phases were combined and evaporated in vacuo to an oily residue.

The crude extract (160 mg) thus obtained was separated on preparative HPLC: the following system was used for this: Hardware Gilson Abimed HPLC (UV detector, binary pump system); Software Unipoint 1.71 (Gilson); Flow 9 ml / min; mobile phase 0.1% TFA (trifluoroacetic acid): ACN (acetonitrile); stationary phase: Merck LichroSorb RP-18 (7 μm; 250 x 25 mm); Gradient: start with t = 0 '(20% ACN); linear to t = 70 '(100% ACN); isocratic up to t = 88 '(100% ACN). Product was eluted from the column with a retention time (R _t ) of 60-64 min (UV absorption at 210 nm).

The yield was 16 mg of the title compound from the mycelium of 1 L culture.

The retention times R _t and ESI data given below relate to the HPLC-MS system described above:

EI-MS: m / z (%) = 189 (100), 191 (64), 193 (10), 450 (26), 452 (38), 454 (21), 456

(6), 656 (16), 658 (32), 660 (26), 662 (11), 664 (2).

C ₂₄ Hι ₄ Cl ₆ O ₉ : ESI-HPLC-MS: R _t = 4.65-4.67 min -ESI neg .: m / z (%) = 654.9 (53),

656.9 (100), 658.9 (86), 660.9 (37), 662.9 (10) C ₂₄ Hι ₄ Cl ₆ O ₉ : HR-MS (EI): calc. 655.8769, found. 655.8782. C ₂₄ Hι ₄ Cl ₆ O ₉ : HR-MS (ESI neg.) From C24Hι ₃ Cl5 ³⁵ Clι ³⁷ O ₉ : calcd. 656.8661, found. 656.8723.

1H-NMR (500 MHz, d ₆ -DMSO) (Fig. 1): δ _H = ca.11.3 (s, broad, 2-3 H, OH), 7.86 (s, 2H, 3'-H and 7 ' -H), 7.81 (s, 4H, 3 "-H and 7" -H), 5.66 (m, IH, 2-H), 4.71 (dd,

J = 12.0, 4.0 Hz, 2H, l _a -H and 3 _a -H), 4.62 (dd, J = 12.0, 6.2 Hz, 2H, l _b -H and 3 _b -H) ¹³ C-NMR (125 MHz , d ₆ -DMSO) (Fig. 2): δ _c = 163.66 (Cl "), 163.41 (C-l '), 154.20 (C-5'), 154.07 (C-5"), 130.03 (C-3 'and C-7'), 129.90 (C-3 "and C-7"), 122.49 (C-4 'and C-6'), 122.43 (C-4 "and C-6"), 121.70 (C -2 'and C-2 "), 70.40 (C-2), 63.48 (Cl and C-3)

Nerfahren 3: Representation from the mycelium of a fermentation on a 30 L scale (MGP medium)

The mycelium from a fermentation in 200 L MGP medium was removed from the supernatant

Centrifugation separated and combined. The moist mycelium was then extracted twice with 5 L acetone for 30 min each in an ultrasonic bath and filtered off. The filtered acetone-water mixture was evaporated in vacuo at approx. 40 ° C. and 250 mbar. The resulting aqueous residue was diluted to 1 L with tap water and then extracted three times with one L each of ethyl acetate. The organic phases were combined and evaporated in vacuo to an oily residue.

The crude extract thus obtained (4.5 g) was distributed in portions of 2.15 g each and separated on preparative HPLC: the following system was used for this:

Hardware Gilson Abimed HPLC (UV detector, binary pump system); Software Unipoint 1.71 (Gilson); Flow 9 ml / min; mobile phase 0.1% TFA (trifluoroacetic acid): ACΝ (acetonitrile); stationary phase: MZ analysis technology (7 μm; 250 x 25 mm); Gradient: start with t = 0 '(20% ACΝ); linear to t = 20 '(45% ACΝ); isocratic up to t = 55 '(45% ACΝ), linear up to t = 90 (100% ACΝ; isocratic to t = 132 '(100% ACN). Agaricoglyceride A was eluted from the column with a retention time (Rt) of 101-108 min.

The total yield was 291 mg of the title compound from the mycelium of 30 L culture.

Nerfahren 4: Representation from the mycelium of a fermentation on a 200 L scale (Q6 medium)

The mycelium from a fermentation in 200 L of Q6 medium was removed from the supernatant

Centrifugation separated and combined. The moist mycelium was then extracted four times with 7 L acetone each for 30 min in an ultrasonic bath and filtered off. The filtered acetone-water mixture was evaporated in vacuo at about 40 ° C and 250 mbar. The resulting aqueous residue was diluted to 1 L with tap water and then extracted three times with one L each of ethyl acetate. The organic phases were combined and evaporated in vacuo to an oily residue.

The crude extract obtained in this way (56 g) was used for the separation of lipophilic components and anti-foam using vacuum column chromatography for the preparative

Pretreated for HPLC. For this purpose, the material to 50 g was C _j g ftmktionalisiertem silica gel (Sigma Aldrich 37.763 to 5) adsorbed. The solvent was evaporated in vacuo and the dry extract / silica gel mixture was applied to a chromatography column consisting of the same cig chromatography material that had been equilibrated with 50% acetonitrile (column size

30 x 4.8 cm). A gradient in 3 stages (200 ml 50%, 80% and 100% aqueous acetonitrile) was used for the crude fractionation of the extract. As shown by an analytical HPLC control, the title compound was enriched in the 80% acetonitrile fraction (intermediate 1). This intermediate product (32 grams) was concentrated on a rotary evaporator at 40 ° C. in vacuo and further separated by preparative HPLC. For this purpose, the oily intermediate 1 produced as described above was divided into aliquots of approx. 3 grams and initially separated on a C8 column from MZ Analysentechnik using the same methodology as described for the purification of the material from the fermentation on a 30 L scale. The

The title compound was highly enriched from the column with a retention time (R _t ) of 99-112 min in an intermediate 2.

This intermediate product 2 (together 3.2 g from 32 grams of intermediate product 1) was portioned into aliquots of approximately 250 mg and again using a Merck

LiChroSorb C18 column purified as in the material from the flask fermentation, the pure title compound eluting at R _t = 59-65 min. The yield was 1.5 grams of agaricoglyceride A from the mycelium of 30 L culture.

An oily intermediate 3 (332 mg) eluted in the HPLC described above

Separation at MZ C8 at R _t = 94-98 min. It was divided into 3 portions xmd each of which was separated on a Waters SymmetryPrep C18 (7 μm, 300 x 19 mm) at a flow of 9 ml / min in the gradient described above. Hardware Gilson Abimed HPLC (UV detector, binary pump system); Software Unipoint 1.71 (Gilson); Flow 9 ml / min; mobile phase 0.1% TFA (trifluoroacetic acid): ACN

(Acetonitrile); stationary phase: a Waters SymmetryPrep C18 (7 μm, 300 x 19 mm); Gradient: start with t = 0 '(50% ACN); isocratic to t = 10 ', linear to t = 25' (80% ACN); linear to t = 44 '(100% ACN). The title compound eluted here at Rt = 30-32 min and Example 2 at R _t = 29-30 min. The yields were 82 mg of the title compound and 1.7 mg of Example 2. Process 5: Increasing the production of agaricoglyceride A by adding 3,5-dichloro-4-hydroxybenzoic acid and 4-hydroxybenzoic acid

Shake flask fermentations of the strain Agaricus sp. WP 4080 was carried out in medium M2 under the same conditions as described above. 1 ml of a solution of 50 mg of 3,5-dichloro-4-hydroxybenzoic acid and 4-hydroxybenzoic acid in GlycerimMeOH 1: 1 was added to the media.

After a fermentation period of up to 300 h, extracts were prepared as described above and compared for agaricoglyceride A content using analytical HPLC as described above, compared. In comparison to the control (conventional MGP medium, 13 mg / 1) there was an increase in production. When 3,5-dichloro-4-hydroxybenzoic acid was added, a content of agarico-glyceride A of 20.4-22.6 mg / 1 was measured, while the content when adding 4-

Hydroxybenzoic acid increased to 18.2 - 19.0 mg.

Example 2

2 - [(3-chloro-4-hydroxybenzoyl) oxy] -l - {[(3,5-dichloro-4-hydroxybenzoyl) oxy] - methyl} ethyl-3,5-dichloro-4-hydroxybenzoate or 2- [ (3-chloro-4-hydroxybenzoyl) - oxy] -3 - [(3,5-dichloro-4-hydroxybenzoyl) oxy] propyl-3,5-dichloro-4-hydroxybenzoate (agaricoglyceride B)

ESI-HPLC-MS: R _t = 4.4 min -ESI neg .: m / z (%) = 621.0 (60), 622.8 (100), 624.7

(95), 626.5 (36).

C ₂₄ Hi ₅ Cl ₅ O ₉ : HR-MS (ESI neg.) Of C ₂₄ Hi ₄ Cl ₅ ³⁵ O ₉ : calc. 620.9080, found. 620.9088.

Examples 3-5

To prepare monoacetylated derivatives, a solution of the compound from Example 1 in abs. Dichloromethane 0.75 eq. Acetic anhydride and pyridine added in excess. The mixture is refluxed. The residue is chromatographed on a reverse phase Cl 8 Aqua HPLC column. A gradient of 0% to 100% aqueous acetonitrile with the addition of 0.1% TFA in 70 min at a flow rate of 7 ml / min is used to remove the products at R _t = approx. 52 min; 20% yield (Example 3) or R _t = approx. 58 min; 4-5% yield (Example 4) or R _t = approx. 63 min; Elute 1% yield (Example 5).

To prepare the fully acetylated derivative, a solution of the compound from Example 1 in abs. Dichloromethane acetic anhydride and pyridine added in excess. The mixture is stirred for 2 hours at room temperature and purified by chromatography using the above method.

Example 3

2,3-bis [(3,5-dichloro-4-hydroxybenzoyl) oxy] propyl 4- (acetyloxy) -3,5-dichlorobenzoate (Example 3a) and 2 - [(3,5-dichloro-4-hydroxybenzoyl ) oxy] -l - {[(3,5-dichloro-4-hydroxybenzoyl) oxy] methyl} ethyl 4- (acetyloxy) -3,5-dichlorobenzoate (Example 3b)

Example 3a Example 3b 1H-NMR of the 2: 1 mixture (500 MHz, d ₆ -DMSO), indication of the integrals relates to the 2: 1 mixture, signal at 5.68 (2-H in both structures) corresponds to 3 H: δ _H = approx. l 1.3 (s, broad, 5-6 H, OH in both structures), 8.06 (s, 2H, 3'-H and 7'-H in Example 3b), 8.02 (s, 4H, 3 "'- H and 7 "'- H in Example 3a), 7.87 (s, 4H, 3" -H, 3 "' - H, 7" -H and 7 "'- H in Example 3b), 7.83 (s, 4H, 3' -H and 7'-H or 3 "-H and 7" -H in Example 3a), 7.82 (s, 4H, 3'-H and 7'-H or 3 "-H and 7" -H in Example 3a ), 5.68 (m, 3H, 2-H in both structures), 4.74 (m, 6H, l _a -H xmd 3 _a -H in both structures), 4.65 (m, 6H, l _b -H and 3 _b - H in both structures), 2.44 (s, 9H, COCH ₃ in both structures).

C ₂₆ Hι ₆ Cl ₆ Oι ₀ : ESI-HPLC-MS: R _t = 5.1 min -ESI neg .: m / z (%) = 697.0 (51), 699.0 (100), 701.0 (83), 703.0 (39 ), 704.9 (11).

Example 4

2 - {[4- (acetyloxy) -3,5-dichlorobenzoyl] oxy} -l - {[(3,5-dichloro-4-hydroxybenzoyl) oxy] - methyl} ethyl-4- (acetyloxy) -3, 5-dichlorobenzoate and 3 - {[4- (acetyloxy) -3,5-dichlorobenzoyl] -oxy} -2 - [(3,5-dichloro-4-hydroxybenzoyl) oxy] propyl-4- (acetyloxy) - 3,5-dichlorobenzoate

Example 4a Example 4b

1H-NMR of the 2: 1 mixture (500 MHz, d ₆ -DMSO), indication of the integrals relates to the 2: 1 mixture, signal at 5.72 (2-H in both suicides) corresponds to 3 H: δκ ^ ca.11.3 (s, broad, 2-3 H, OH in both structures), 8.06 (s, 4H, 3 "-H, 7" -H, 3 "'- H and 7"' - H in Example 4b), 8.01 ( s, 8H, 3'-H, 7'-H, 3 "'- H and 7"' - H in Example 4a), 7.86 (s, 2H, 3'-H and 7'-H in Example 4b), 7.81 (s, 4H, 3 "-H and 7" -H in Example 4a), 5.72 (m, 3H, 2-H in both structures), 4.78 (m, 6H, l _a -H and 3 _a -H in both structures), 4.68 (m, 6H, l _b -H and 3 _b -H in both structures), 2.43 ( s, 18H, COCH ₃ in both structures).

C ₂₈ Hι ₈ Cl ₆ On: ESI-HPLC-MS: R _t = 5.5 min -ESI neg .: m / z (%) = 739.0 (49), 741.0 (100), 743.0 (76), 745.0 (38) , 747.0 (11), 749.0 (2)

Example 5

2 - {[4- (acetyloxy) -3,5-dichlorobenzoyl] oxy} -l - ({[4- (acetyloxy) -3,5-dichlorobenzoyl] - oxy} methyl) ethyl-4- (acetyloxy) -3 , 5 -dichlorobenzoate

ESI-HPLC-MS: R _t = 5.87 min

1H-NMR (500 MHz, d ₆ -DMSO): δ _H = 8.10 (s, 2H, 3'-H and 7'-H), 8.06 (s, 4H, 3 "-H and 7" -H), 5.74 (m, IH, 2-H), 4.81 (dd, J = 12.1, 4.0 Hz, 2H, l _a -H and 3 _a -H), 4.71 (dd, J = 12.1, 6.2 Hz, 2H, l _b -H and 3 _b -H), 2.45 (s, 9H, COCH ₃ ).

The examples listed in Table 3 can be prepared from the corresponding starting compounds analogously to the instructions in Example 1:

Example 13

2 - [(5-Chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] -l - {[(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] methyl} ethyl-4- (benzyloxy ) -5-chloro-2,3-difluoro-benzoate

2 - [(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] -l - {[(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] methyl} ethyl-4- (Benzyloxy) -5-chloro-2,3-difluorobenzoate could be used in the

Preparation of Example 10 can be isolated as a by-product.

MS (ESIpos): m / z = 770 (M + NH ₄ ) ⁺ ;

HPLC (method A): R _t = 5.35 min;

1H-NMR (200 MHz, CDC1 ₃ ): δ - 7.82-7.71 (m, 3H), 7.49-7.31 (m, 5H), 5.72 (m,

IH), 5.32 (s, 2H), 4.83-4.52 (m, 4H).

The examples listed in Table 4 can be prepared analogously to the synthesis sequence of Example 1 with the corresponding diols as starting compounds (instead of glycerol):

Example 31

2- [(3, 5-dichloro-4-hydroxybenzoyl) oxy] - 1 - {[(3, 5-dichloro-4-methoxybenzoyl) oxy] - methyl} ethyl-3, 5-dichloro-4-hydroxybenzoate

Example 31 can be prepared analogously to example 39 from the corresponding starting compounds. MS (ESIneg): mz = 683 (MH) ^" ; LC / MS (Method 1): R _t = 7.1 min;

1H-NMR (500 MHz, d ₆ -DMSO): δ = 11.3 (s, broad, IH), 7.95 (s, 2H), 7.88 (s, 2H), 7.78 (s, 2H), 5.69 (m, IH ), 4.77-4.63 (m, 4H), 3.88 (s, 3H), 3.87 (s, 3H).

The examples listed in Table 5 can by acylation of Example 2A with appropriate acid chlorides, followed by hydrogenolytic deprotection of the

Phenols can be prepared analogously to the synthesis of Example 1:

Example 35

2- (Benzoyloxy) -3 - [(3-chloro-4-hydroxybenzoyl) oxy] propyl-3,5-dichloro-4-hydroxybenzoate

2- (Benzoyloxy) -3 - [(3-chloro-4-hydroxybenzoyl) oxy] propyl-3,5-dichloro-4-hydroxybenzoate could be isolated as a by-product in the preparation of Example 34.

MS (ESIneg): m / z = 537 (MH) ^" ;

LC / MS (Method 1): R _t = 6.2 min;

1H-NMR (500 MHz, d ₆ -DMSO): δ = 11.2 (s, 2H), 7.96 (d, 2H), 7.83 (m, IH), 7.81

(s, 2H), 7.75 (m, IH), 7.66 (dd, IH), 7.52 (dd, 2H), 7.04 (d, IH), 5.72 (m, IH), 4.73 -

4.66 (m, 2H), 4.64-4.57 (m, 2H).

Example 36

2- {4 - [(Benzyloxy) carbonyl] phenoxy} -3 - [(3,5-dichloro-4-hydroxybenzoyl) oxy] propyl-3, 5-dichloro-4-hydroxybenzoate

130 mg (0.2 mmol) of Example 2A, 50.0 mg (0.22 mmol) of benzyl p-hydroxybenzoate and 67.9 mg (0.26 mmol) of triphenylphosphine are successively dissolved in 5 ml of abs. Dissolved tetrahydrofuran and stirred at 0 ° C for ten minutes. Then 34.5 μl (0.22 mmol) of diethyl azodicarboxylate are added dropwise. The reaction mixture is warmed to RT in an ice bath, stirred for 3 days and then concentrated. The residue is drawn up on silica gel 60 and purified on silica gel 60 (mobile phase: cyclohexa ethyl acetate 50: 1, 20: 1, 10: 1, 2.1, 1: 1). 116 mg (68% of theory) of 2- {4 - [(benzyloxy) carbonyl] phenoxy} -3 - {[4- (benzyloxy) -3,5-dichlorobenzoyl] oxy} propyl-4- (benzyloxy) are obtained ) -3,5-dichlorobenzoate, which analogously to Example 1 to the product in a yield of 51% of theory. Th. Is deprotected. MS (ESIneg): m / z = 681 (MH) ^"

Example 37

3 - [(3,5-dichloro-4-hydroxybenzoyl) oxy] -2- [3 - (trifluoromethyl) phenoxy] propyl-3,5-dichloro-4-hydroxybenzoate

Example 37 can be produced analogously to example 36 from the corresponding starting compounds.

MS (ESIneg): m / z = 611 (MH) ^" ; HPLC (method A): R _t = 5.27 min. Example 38

2 - [(3,5-dichloro-4-methoxybenzoyl) oxy] -l - {[(3,5-dichloro-4-methoxybenzoyl) - oxy] methyl} ethyl 3,5-dichloro-4-hydroxybenzoate

Example 38 can be prepared analogously to example 32 from the corresponding starting compounds. MS (ESIneg): m / z = 687 (MH) ^" ;

1H-NMR (500 MHz, d ₆ -DMSO): δ = approx.11.3 (s, broad, 1 H), 7.88 (s, 4 H), 7.85 (s, 2 H), 5.68 (m, 1 H) , 4.78-4.63 (m, 4H), 3.87 (s, 6H).

Example 39

2,3-bis [(3,5-dichlorobenzoyl) oxy] propyl-3,5-dichloro-4-hydroxybenzoate

Example 39 can be prepared by reacting 2 - [(3,5-dichlorobenzoyl) oxy] -1- (hydroxymethyl) ethyl-3,5-dichlorobenzoate (Example 3A) with 4- (benzyloxy) -3,5-dichlorobenzo- Acid chloride (Example 12A) can be obtained analogously to Example 1. 1H-NMR (500 MHz, d ₆ -DMSO): δ = ca.ll.3 (s, broad, OH), 7.95 (s, IH), 7.93 (s, IH), 7.89 (s, 2H), 7.83 (s, 2H), 7.81 (s, 2H), 5.72 (m, IH), 4.79-4.63 (m, 4H). Example 40

2 - [(3,5-dichloro-4-hydroxybenzoyl) oxy] -l - {[(3,5-dichloro-4-methoxybenzoyl) oxy] methyl} ethyl 3,5-dichloro-4-hydroxybenzoate

Example 40 can be prepared analogously to example 39 from the corresponding starting compounds. MS (ESIneg): m / z = 669 (MH) ^" ; LC / MS (method 1): R _t = 6.6 min;

1H-NMR (500 MHz, d ₆ -DMSO): δ = 11.3 (s, broad, 2 H), 7.89 (s, 2H), 7.86 (s, 2H), 7.79 (s, 2H), 5.66 (m, IH), 4.75-4.62 (m, 4H), 3.89 (s, 3H).

-1 Form - PCT / RO / 134 (EASY) Information on a deposited microorganism and / or other deposited biological material -1-1 created by using PCT-EASY version 2. 92 (updated 01.06.2002) -2 international file number.

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INTERNATIONAL FORM

BAYER AG Aprat er Weg 18 a

FIRST DEPOSIT RECEIPT CONFIRMATION, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

MUCL 29004

DSM 14896

H. SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

(X) submitted a proposed taxonomic designation (check where applicable).

HI. ENTRANCE AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-03-08 (date of first deposit) ¹

IV. RECEIPT FOR CONVERSION

The microorganism referred to in 1 has been received by this international depository on (date of first filing) and an application for conversion of this first deposit into a deposit under the Budapest Treaty has been received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF SIGNATURES) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by them:

Address: Mascheroder Weg lb D-38124 Braunschwei ^' g

Date: 2002-04-04

¹ If Rule 6.4 (d) applies, this is the time at which the status of an international depositary was acquired. Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

RECOGNITION OF THE DEPOSIT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERMEDIATE FORM

BAYER AG Aprather Weg 18 a

RECEIPT CONFIRMATION ON FIRST DEPOSIT, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

MUCL 28242

DSM 14897

E SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

(X) submitted a proposed taxonomic name. (Tick the appropriate).

HL INPUT AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-03-08 (date of first deposit).

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I was received by this international depository on (date of first deposit) and an application for the conversion of this first deposit into a deposit under the Budapest Treaty was received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Surname; DSMZ-GERMAN COLLECTION OF Signature (s) of the person (s) authorized to represent the international depository parts MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by them:

Address: Mascheroder Weg lb. D-38124 Braunschweig

LZ ^, C ^^ ^' ^

Date: 2002-04-04

¹ If Rule 6.4 (d) applies, this is the time at which the status of an international depositary was acquired. Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

RECOGNITION OF THE DEPOSIT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprather Weg 18 a

FIRST DEPOSIT RECEIPT CONFIRMATION, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

I. LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

FU41954

DSM 14898

IL SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

() submitted a proposed taxonomic name (check where applicable).

HI. ENTRANCE AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-03-08 (date of first deposit) ¹ .

IV, RECEIPT OF CONVERSION APPLICATION

The microorganism referred to under I was received by this international depository on (date of first deposit) and an application for the conversion of this first deposit into a deposit under the Budapest Treaty was received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF SIGNATURES) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by them:

Address: Mascheroder Weg lb. D-38124 Braunschweig

Date: 2002-04-04

¹ If Rule 6.4 (d) applies, this is the time at which the status of an international depositary was acquired. Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

RECOGNITION OF THE DEPOSIT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprather Weg 18 a

FIRST DEPOSIT RECEIPT CONFIRMATION, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT: WP4121

DSM 14899

π. SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

(X) submitted a proposed taxonomic name. (Tick the appropriate).

DT. ENTRANCE AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-03-08 (date of first deposit)

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I has been received by this International Depository on (date of first filing) and an application for conversion of this first deposit into a deposit under the Budapest Treaty has been received on (date of receipt of the application for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF SIGNATURES) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by them:

Address: Mascheroder Weg lb. D-38124 Braunschweig

Date: 2002-04-04

¹ If Rule 6.4 Book d applies, this is the time at which the status of an international depository was acquired Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

RECOGNITION OF THE DEPOSIT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprather Weg 18 a

RECEIPT CONFIRMATION ON FIRST DEPOSIT, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

WP0058

DSM 14900

IL SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC REFERENCES

With the microorganism designated under I.

() a scientific description

() submitted a proposed taxonomic name. (Tick the appropriate).

HL INPUT AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-03-18 (date of first deposit) ¹

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I has been received by this International Depositary on (date of first deposit) and an application for the conversion of this first deposit into a deposit under the Budapest Treaty has been received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF Signature (s) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by her:

Address Mascheroder Weg lb D-38124 Brauπschwe-g cx. cx _* .x _*

Date: 2002-04-04

¹ If rule 6.4 letter d applies, this is the time at which the status of an international depository was acquired Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

ACKNOWLEDGMENT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprather Weg 18 a

FIRST DEPOSIT RECEIPT CONFIRMATION, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

WP0059

DSM 14901

IL SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

() submitted a proposed taxonomic term. (Tick the appropriate).

EL INPUT AND ACCEPTANCE

This international depository accepts the microorganism designated under I, which it received on 2002-03-18 (date of first filing).

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I was received by this international depository on (date of first deposit) and an application for the conversion of this first deposit into a deposit under the Budapest Treaty was received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF Signature (s) of the person (eπ) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by her:

Address: Mascheroder Weg lb

D-38124 Braunschweig X- AXCAX,

Date: 2002-04-04

If rule 6.4 letter d applies, this is the time at which the status of an international depository was acquired Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

RECOGNITION OF THE DEPOSIT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprather Weg 18 a

FIRST DEPOSIT RECEIPT CONFIRMATION, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

I. LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

WP0360

DSM 14902

EL SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

(X) submitted a proposed taxonomic name. (Tick the appropriate).

HL INPUT AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-03-18 (date of first deposit) ¹ .

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I was received by this international depository on (date of first deposit) and an application for the conversion of this first deposit into a deposit under the Budapest Treaty was received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF Untersehrift (s) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by them:

Address: Mascheroder Weg lb

D-381 4 Braunschweig

Date: 2002-04-04

If Rule 6.4 (d) applies, this is the time at which the status of an international depository was acquired Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

RECOGNITION EGG DEPOSIT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprather Weg 18 a

FIRST DEPOSIT RECEIPT CONFIRMATION, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

FU40473

DSM 14903

H. SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

() submitted a proposed taxonomic name (check where applicable).

HL INPUT AND ACCEPTANCE

This international depository accepts the microorganism designated under I, which it received on 2002-03-18 (date of first deposit) ¹ .

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I was received by this international depository on (date of first deposit) and an application for the conversion of this first deposit into a deposit under the Budapest Treaty was received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF SIGNATURES) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by them:

Address: Mascheroder Weg Ib D-38124 Braunschweig IX. ^ -

Date: 2002-04-04

¹ If rule 6.4 letter d applies, this is the point in time at which the status of an internal depository was acquired. Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

ACKNOWLEDGMENT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprather Weg 18 a

RECEIPT CONFIRMATION ON FIRST DEPOSIT, issued in accordance with Rule 7.1 by the INTERNATIONAL DEPOSITOR below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT:

DSM 11373

DSM 14904

IL SCIENCE TOWEL DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

(X) submitted a proposed taxonomic name. (Tick the appropriate).

H\. ENTRANCE AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-03-18 (date of first deposit) ¹ .

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I was received by this international depository on (date of first deposit) and an application for the conversion of this first deposit into a deposit under the Budapest Treaty was received on (date of receipt of the request for conversion).

V. INTERNATIONAL DEPOSIT

Name: DSMZ-GERMAN COLLECTION OF SIGNATURES) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by them:

Address: Mascheroder Weg lb D-3S124 Braunschweig

Date: 2002-04-04

¹ If Rule 6.4 letter d applies, this is the time at which the status of an international depository was acquired Form DSMZ-BP / 4 (single page) 12/2001 BUDAPEST AGREEMENT ON THE INTERNATIONAL

ACKNOWLEDGMENT OF MICROORGANISMS

FOR THE PURPOSES OF PATENT PROCEDURES

INTERNATIONAL FORM

BAYER AG Aprat erWeg 18 a

RECEIPT CONFIRMATION AT THE FIRST DEPOSIT, issued in accordance with regulations! 7.1 from the INTERNATIONAL DEPOSITER indicated below

42096 Wuppertal

L LABELING OF THE MICROORGANISM

Reference number assigned by the DEPOSIT: INPUT NUMBER assigned by the INTERNATIONAL DEPOSIT: CBS585.76

DSM 14921

H. SCIENTIFIC DESCRIPTION AND / OR PROPOSED TAXONOMIC NAME

With the microorganism designated under I.

() a scientific description

(X) submitted a proposed taxonomic name (check where applicable).

DL INPUT AND ACCEPTANCE

This international depository accepts the microorganism referred to under I, which it received on 2002-04-08 (date of first deposit)

IV. RECEIPT FOR CONVERSION

The microorganism referred to under I was received by this international depository on (date of first deposit) and an application for the conversion of this first deposit into a deposit pursuant to the Budapest Treaty was received on (Daturii of receipt of the application for conversion).

V. I-TRERNATIONAL REAR - LOCATION

Name: DSMZ-GERMAN COLLECTION OF Signature (s) of the person (s) authorized to represent the international depositary MIKROORGANISMEN UND ZELLKULTUREN GmbH or the employee (s) authorized by her:

Address: Mascheroder Weg lb D-38124 Brat-nschwetg

Date: 2002-04-16

¹ If rule 6.4 letter d applies, this is the point in time at which the status of an international depositary has been αworbeπ. Form DSMZ-BP / 4 (single page) 12/2001

Claims

claims

1. Compounds of the formula

in which

E for a remainder of the formula

stands,

R, R, R, R, R and R independently of one another are radicals selected from the group consisting of hydrogen, halogen, cyano, C 1 -C ₄ -alkyl and C 1 -C ₄ alkoxy,

where at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} halogen or cyano,

R ² , R ⁵ and R ⁸ independently of one another for radicals selected from the group hydrogen, hydroxy, acetoxy, amino, carboxyl, halogen,

Cyano and -CC-alkyl stand, where at least one of the radicals R ² , R ⁵ and R ^{8 is} hydroxy, acetoxy, amino or carboxyl,

and

where R ² , R ⁵ and R ^{8 are} each in the meta or para position of the benzoyl radicals,

R .1 ^ι 0 ^{υ represents} hydrogen or Cι-C ₆ alkyl,

or

stands for -CRnR.12-,

R ^{1 represents} halogen or cyano,

R ^{2 represents} hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another represent radicals selected from the group consisting of hydrogen, halogen and cyano,

R ^{8 represents} hydrogen, hydroxy or acetoxy,

and

where R ² and R ^{8 are} each in the meta or para position of the benzoyl radicals,

R ^{11 represents} hydrogen or CC ₄ alkyl,

R ¹² is C _ö -Cio-aryl or 5- to 10-membered heteroaryl, where C ₆ -C ₁₀ aryl and 5- to 10-membered heteroaryl with up to 3 substituents independently selected from the group halogen, cyano, trifluoromethyl, Cr alkyl and C ₁ -C ₆ alkoxy may be substituted,

or

R and R together with the carbon atom to which they are attached form a 4- to 6-membered cycloalkyl ring,

or

E for a remainder of the formula

stands,

R represents halogen or cyano,

R ^{2 represents} hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another represent radicals selected from the group consisting of hydrogen, halogen, C ₁ -C alkyl and cyano,

R ^{8 represents} hydrogen, hydroxy or acetoxy, and

where R ² and R ^{8 are} each in the meta or para position of the benzoyl radicals,

R, 1 ^ι 0 ^{υ represents} hydrogen or Cι-C ₆ alkyl,

R ^{13 represents} phenyl,

where phenyl can be substituted with up to 3 substituents independently of one another selected from the group consisting of halogen, cyano, trifluoromethyl, C ₁ -C ₆ -alkyl, C Cό-alkoxy and -CO ₂ R ¹⁵ ,

where R ^{15 is} -C ₆ alkyl and benzyl,

and their solvates of salts.

Compounds according to claim 1, of the formula

in which R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ⁹ independently of one another are radicals selected from the group consisting of hydrogen, halogen, cyano, C ₁ -C ₄ alkyl and C ₁ -C ₄ alkoxy,

where at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} halogen or cyano,

R ² , R ⁵ and R ⁸ independently of one another are radicals selected from the group consisting of hydrogen, hydroxy, acetoxy, amino, carboxyl, halogen, cyano and C 1 -C ₄ -alkyl,

where at least one of the radicals R, R and R is hydroxyl, acetoxy, amino or carboxyl,

and

where R ² , R ⁵ and R ^{8 are} each in the meta or para position of the benzoyl radicals,

and

R ^{10 represents} hydrogen or -CC ₆ alkyl,

and their salts, solvates and solvates of the salts.

3. Compounds according to claim 1, wherein

R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ⁹ independently of one another represent hydrogen or chlorine, where at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} chlorine,

R ² , R ⁵ and R ⁸ independently of one another for radicals selected from the group

Are hydrogen, hydroxy and acetoxy,

wherein at least one of the radicals R, R and R is hydroxy or

Acetoxy means

and

where R _> 2, r R5 and R are each in the para position of the benzoyl radicals,

and

R ^{10 represents} hydrogen,

and their salts, solvates and / or solvates of the salts.

4. Compounds according to claim 1, wherein

R, R, R, R, R and R are selected independently of one another from the group hydrogen and chlorine,

where at least one of the radicals R ¹ , R ³ , R ⁴ , R ⁶ , R ⁷ and R ^{9 is} chlorine,

and R ² , R ⁵ , R ⁸ and R ^{10 have} the meanings given above.

5. Compounds according to claim 1, in which

E stands for -CR11R12-,

R ^{1 represents} halogen or cyano,

R ^{2 represents} hydroxy or acetoxy, "in O

R, R and R independently of one another for radicals selected from the group

Represent hydrogen, halogen and cyano, R ^{8 represents} hydrogen, hydroxy or acetoxy,

and

9 R where R and R are each in the meta or para position of the benzoyl radicals,

R ^{11 represents} hydrogen or -CC ₄ alkyl,

R ^{12 represents} C ₆ -Cιo-aryl or 5- to 10-membered heteroaryl,

wherein C ₆ -Cι ₀ aryl, and 5- to 10-membered heteroaryl having up to

3 substituents selected independently of one another from the group halogen, cyano, trifluoromethyl, C ₁ -C ₆ -alkyl and C ₁ -C ₆ alkoxy may be substituted,

or

R ^π and R ¹² together with the carbon atom to which they are attached form a 4- to 6-membered cycloalkyl ring,

and their salts, solvates and / or solvates of the salts.

6. Compounds according to claim 1, in which

E stands for -CRnRn-,

R ¹ , R ³ , R ⁷ and R ^{9 represent} chlorine, where R, R, R and R are each in the meta position of the benzoyl radicals,

9 o

R and R represent hydroxy,

9 R where R and R are each in the para position of the benzoyl radicals,

R ^{11 represents} hydrogen or methyl,

and

R ^{12 represents} phenyl, pyridyl or quinolinyl,

where phenyl can be substituted once with methoxy,

and their salts, solvates and or solvates of the salts.

7. Compounds according to claim 1, wherein

E for a rest of the. formula

ooddeerr stands,

R ^{1 represents} halogen or cyano,

R ^{2 represents} hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another for radicals selected from the group

Are hydrogen, halogen, -CC ₄ -alkyl and cyano,

R ^{8 represents} hydrogen, hydroxy or acetoxy, and

where R ² and R ^{8 are} each in the meta or para position of the benzoyl radicals, R ^{10 is} hydrogen or -CC ₆ alkyl,

and

R ^{13 represents} phenyl,

where phenyl with up to 3 substituents independently selected from the group halogen, cyano, trifluoromethyl, C _t -C _ö alkyl, -CC ₆ alkoxy and -CO ₂ R ¹⁵ can be substituted,

where R ^{15 is} -C ₆ alkyl and benzyl,

and their salts, solvates and / or solvates of the salts.

8. Compounds according to claim 1, in which

E for a remainder of the formula

or stands

R ¹ , R ³ , R ⁷ and R ^{9 represent} chlorine,

where R ¹ , R ³ , R ⁷ and R ^{9 are} each in the meta position of the benzoyl radicals,

R ² and R ^{8 represent} hydroxy, where R .2 - un-. d R are each in the para position of the benzoyl radicals,

R, 14 represents hydrogen,

and

R represents phenyl,

where phenyl can be substituted with up to 3 substituents independently of one another selected from the group consisting of chlorine, fluorine, bromine, trifluoromethyl, -CC alkyl, methoxy and -CO ₂ benzyl.

2 - [(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] -l - {[(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] methyl} ethyl-4- ( benzyloxy) -5-chloro-2,3-difluorobenzoate

2,3-bis [(5-chloro-2,3-difluoro-4-hydroxybenzoyl) oxy] propyl 5-chloro-2,3-difluoro-4-hydroxybenzoate

2- [(3,5-dichloro-4-hydroxybenzoyl) amino] -1 - {[(3,5-dichloro-4-hydroxybenzoyl) oxy] methyl} ethyl-3,5,5-dichloro-4-hydroxybenzoate

and their salts, solvates and solvates of the salts.

10. Use of compounds of formula (I) in which

stands for - (CH ₂ ) _n - or -CR ₁₆ Rι ₇ -,

R ^{1 represents} halogen or cyano,

R) 2 represents hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another represent radicals selected from the group consisting of hydrogen, halogen and cyano, R ^{8 represents} hydrogen, hydroxy or acetoxy,

and

9 R where R and R are each in the meta or para position of the benzoyl radicals,

n stands for 0 to 3,

R ^{16 represents} hydrogen, -CC ₆ -alkyl or C ₃ -C ₈ -cycloalkyl,

and

R ^{17 represents} Ci-Cβ-alkyl or C ₃ -C ₈ cycloalkyl,

where CC _ö alkyl may optionally be substituted with phenyl,

and their salts, solvates and / or solvates of the salts for the manufacture of a medicament.

11. Use of compounds according to claim 10,

in which

E represents - (CH ₂ ) _n - or -CR ₁₆ R ₁₇ -,

R ^{1 represents} chlorine,

R ^{2 represents} hydroxy or acetoxy,

R ³ , R ⁷ and R ⁹ independently of one another represent hydrogen or chlorine, R ^{8 represents} hydrogen, hydroxy or acetoxy,

xmd

9 R where R and R are each in the para position of the benzoyl radicals,

n represents 1 to 3,

R ^{16 represents} hydrogen or methyl,

xmd

R ¹⁷ represents C 1 -C ₄ -alkyl,

where C 1 -C ₄ -alkyl may optionally be substituted with phenyl,

and their salts, solvates and / or solvates of the salts for the manufacture of a medicament.

12. A process for the preparation of compounds of formula (I), characterized in that

[A] Compounds of the formula

in which R ¹ to R ³ , R ⁷ to R ⁹ and R ^{10 have} the meanings given in Claim 1,

with compounds of the formula

in which

R, 4, R and R have the meanings given in claim 1,

and

X ^{1 represents} halogen, preferably bromine or chlorine, or hydroxy,

to compounds of the formula

in which

R ¹ to R ^{10 have} the meanings given in claim 1, be implemented

or

[B] Compounds of the formula

in which

R ¹ to R ^{3 have} the meanings given in claim 1,

E for -CRiiRn-, - (CH ₂ ) _n -, -CR ₁₆ Rι ₇ -

or a residue of the formula

stands, and

n, R ¹⁰ to R ¹³ , R ¹⁶ and R ^{17 have} the meanings given in Claim 1,

with preferably one equivalent of a compound of the formula

in which

R> 7, τ R> 8 - u, "nd J R> 9 have the meanings given in Claim 1,

and

X represents halogen, preferably bromine or chlorine, or hydroxy,

to compounds of the formula

in which

for -CR11R12-, - (CH ₂ ) „-, -CRι ₆ R ₁₇ -

or a residue of the formula

stands,

and n, R ¹ to R ³ , R ⁷ to R ¹³ , R ¹⁶ and R ^{17 have} the meanings given in Claim 1,

be implemented

or

[C] Compounds of the formula (II) with compounds of the formula

R ¹³ -OH (VI),

in which

R 1 "^ has the meanings given in claim 1,

to compounds of the formula

in which

1 1 7 10 \ "X

R to R, R to R and R have the meanings given in claim 1,

x be implemented and the resulting compounds (Ib), (Ic) xmd (Id) are optionally reacted with the corresponding (a) solvents and / or (b) bases or acids to give their solvates, salts or solvates of the salts.

13. The method according to claim 12, characterized in that compounds of the formula

in which

R, ι to R, R to R and R, 10 have the meaning given in claim 1,

with compounds of the formula

in which

R 4, R and R have the meaning given in claim 1,

and

X ^{1 represents} halogen or hydroxy, in inert solvents to give compounds of formula (I).

14. Process for the preparation of the compounds of formula

in which

R ^{2 represents} hydroxy or acetoxy,

R ⁶ xmd R ⁹ independently of one another represent hydrogen or chlorine,

characterized in that the compounds of formula (Ia) by fermentation of microorganisms and the subsequent isolation of the

Compound can be obtained from the culture broth.

15. The method according to claim 14, wherein the microorganism is the microorganism with the deposit no. DSM 14593, DSM 14594, DSM 14901, DSM 14894, DSM 14895, DSM 14898, DSM 14899, DSM 14900,

DSM 14892, DSM 14893, DSM 14896, DSM 14897, DSM 14902, DSM 14903, DSM 14904 or DSM 14921.

16. Microorganisms with the deposit no. DSM 14593, DSM 14594, DSM 14901, DSM 14894, DSM 14895, DSM 14898, DSM 14899 and DSM

14,900th

17. Compounds according to any one of claims 1 to 9 for the treatment xmd / or prophylaxis of diseases.

18. Medicament containing at least one of the compounds according to one of the

Claims 1 to 9 in admixture with at least one pharmaceutically acceptable, essentially non-toxic carrier or excipient.

19. Use of compounds according to any one of claims 1 to 11 for the manufacture of a medicament for the treatment and / or prophylaxis of

Pain.

20. Medicament according to claim 18 for the treatment and / or prophylaxis of painful conditions.