WO1998024751A1 - Unsaturated aliphatic carboxylic acids (maracenines), method for the production thereof and agent against bacteria - Google Patents

Abstract

The invention relates to unsaturated aliphatic carboxylic acids (maracenines) of general formula (I), wherein, if x = 2, y = 1 and (a) means a double bond, z = 0, R is missing and (b) is a triple bond or z = 1, R is a C1 atom and (b) is a double bond or if x = 3, y = 2 and (a) is a single bond, z = 0, R is missing and (b) is a triple bond or z = 1, R is a C1 atom and (b) is a double bond. Another embodiment of the invention relates to an agent against bacteria, in particular against mycobacteria and for Nocardia, containing an amount of an unsaturated aliphatic carboxylic acid according to the invention (maracenine) in addition to the usual auxiliary or carrier substances. The agent according to the invention can be intended for the treatment of tuberculosis.

Description

UNSATURATED ALIPHATIC CARBONIC ACIDS (MARACENINE), PRODUCTION PROCESS AND AGENTS AGAINST BACTERIA

The invention relates to unsaturated aliphatic carboxylic acids (maracenins), processes for their production and agents containing unsaturated aliphatic carboxylic acids (maracenins).

According to one embodiment, the invention relates to unsaturated aliphatic carboxylic acids (maracenins) of the general formula:

20 CH _χ - 'CHy-CH = CH-CH2-CH = CH-CH2-CH = CH-CH2-CH2-CH _z == CR-0-CH = CH-CH ₂ -

CH ₂ -C0 ₂ H

where if x = 2, y = 1 and - "- a double bond, z = 0, R is absent and = '= a triple bond or z = 1, R is a Cl atom and == a double bond or, - if x = 3, y = 2 and - 'a single bond, z = 0, R is absent and == a triple bond or z = 1, R is a Cl atom and == a double bond.

The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin Aτ_) of the formula

Maracenin A, delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -trans, delta ¹⁷ ' ¹⁸ - cis and / or with the following parameters:

MG 300.40

¹ H-NMR according to Table 1, ¹³ C-NMR according to Table 1,

IR (KBr); ny = 3014 (w), 2925 (m), 2855 (w), 2280 (m), 1712 (s), 1673 (m), 1662 (m), 1554 (w), 1434 (m), 1412 (m ), 1351 (w), 1222 (s), 1175 (w), 1059 (m), 998 (m), 968 (m) and 920 (m) cm ^"1 ; and UV (methanol); lamda _max (log epsilon) = 208 (4.41) and 227 (4.47) nm.

The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin A2) of the formula

20 19 18 17 15 1 12 11 5 1

CH ₂ = CH-CH = CH-CH ₂ -CH = CH- ^C H ₂ -CH = CH-CH ₂ CH ₂ -C = C-0-CH = CH-CH ₂ CH ₂ -COOH

delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -cis, delta ¹⁷ ' ¹⁸ -cis and / or with the following parameters:

MG 300.40

¹ H NMR (CDCI3, 300 MHz); delta = 2.96 (m, 2H, 15-H) and 2.82 (m,

2H, 12 -H) ppm, and ¹³ C NMR (CDCI3, 75.5 MHz); delta = 26.1 (C-15) and 25.8 (C-12) ppm.

The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin B ^) of the formula:

Maracenin B, 19.20-dihydro, delta ⁴ ' ⁵ -trans, delta ^{11, 12} -trans, delta ¹⁴ ' ¹⁵ - trans, delta ^{7 1} -eis and / or with the following parameters:

MG 302.41,

¹ H-NMR according to Table 2,

¹³ C-NMR according to Table 2,

IR (KBr); ny = 3012 (m), 2964 (m), 2931 (m), 2875 (w), 2279 (m),

1712 (s), 1674 (m), 1662 (m), 1437 (m), 1412 (w), 1304 (w), 1288 (m),

1223 (s), 1061 (m), 968 (m) and 921 (m) cm ^-1 , and

UV (methanol); lamdam x (log epsilon) = 205 (4.37) nm.

The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin B2) of the formula:

20 19 18 17 15 14 12 11 5 4 1

CH3-CH-2CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -C≡C-0-CH = CH-CH ₂ CH ₂ -COOH

19.20-dihydro, delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -cis, delta' -eis and / or with the following parameters:

MG 302.41,

^ H NMR (CDC1 ₃ , 400 MHz); delta = 2.81 (m, 4H, 12- and 15-H) and

¹³ C NMR (CDCI3, 75.5 MHz); delta = 25.8 (C-12) and 25.6 (C-15) ppm. The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin B3) of the formula:

20 19 18 17 15 14 12 11 5 4 1

CH-CH-CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -C = C-0-CH = CH-CH ₂ CH ₂ -COOH

19.20-dihydro, delta ⁴ ' ⁵ -cis, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -trans, delta' -eis and / or with the following parameters:

MG 302.41 and

¹ H NMR (CDC1 ₃ , 400 MHz); delta = 6.21 (d, J = 5.7 Hz, 1H, 5-H) and 4.75 (m, 1H, 4-H) ppm.

The invention further relates to an unsaturated aliphatic carboxylic acid of the formula (Maracenin C ^):

20 19 18 17 15 14 12 11 5 4 1

CH ₂ = CH-CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -CH = C-0-CH = CH-CH ₂ CH ₂ -COOH

Cl delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -trans, delta ¹⁷ ' ¹⁸ - cis and / or with the following parameters:

MG 336.86.

¹ H-NMR according to Table 1, ¹³ C-NMR according to Table 1,

IR (KBr); ny = 3012 (w), 2925 (m), 285 (w), 1711 (s), 1654 (n), 1434 (m), 1412 (w), 1271 (m), 1215 (m), 1154 (s ), 1069 (w), 997 (m), 967 (m) and 924 (m) cm ^-1 , and

UV (methanol); lamda _max (log epsilon) = 205 (4.25) and 224 (4.22) nm.

The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin C) of the formula:

20 19 18 17 15 14 12 11 5 4 1

CH ₂ = CH-CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -CH = C-0-CH = CH-CH ₂ CH ₂ -COOH

Cl delta ^{4, 5} -trans, delta ^{11 '12} -trans, delta ¹⁴ ' ¹⁵ -cis, delta ^{17 '18} -cis and / or with the following parameters:

MG 336.86 and

^ ^• H NMR (CDC1 ₃ , 300 MHz); delta = 2.95 (m, 2H, 15-H) and 2.81 (m,

2H, 12-H) ppm, and

¹³ C NMR (CDCI3, 75.5 MHz); delta = 26.6 (C-15) and 26.1 (C-12) ppm.

The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin D _{j of} the formula:

20 18 17 15 14 12 11

CH ₃ CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH2-CH = C-0-CH = CH-CH ₂ CH ₂ -COOH

I Cl

19.20-dihydro, delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} - trans, delta ^{17, 18} -eis and / or with the following parameters:

MG 338.88,

^ ^• H-NMR according to Table 2,

¹³ C-NMR according to Table 2,

IR (KBr); ny = 3011 (m), 2963 (m), 2929 (m), 2873 (w), 2856 (w),

1713 (m), 1654 (s), 1446 (), 1409 (m), 1270 (), 1155 (s), 1069 (w),

967 (m) and 924 (m) cm ^-1 , and

UV (methanol); lamda _max (log epsilon) = 203 (4.16) nm.

The invention further relates to an unsaturated aliphatic carboxylic acid (maracenin D2) of the formula:

19,20-dihydro, delta ^{4 '5} trans, delta ^{11' 12} trans, delta ^{14 '15} -cis, delta ^{17' 18} cis and / or with the following parameters: MG 338.88,

¹ H NMR (CDCI3, 300 MHz); delta = 2.80 (m, 4H, 12- and 15-H) ppm, and

¹³ C NMR (CDC1 ₃ , 75.5 MHz); delta = 25.7 (C-12) and 25.6 (C-15) ppm.

According to a further embodiment, the invention relates to a process for the production of unsaturated aliphatic carboxylic acids (maracenins), characterized in that

(a) Sorangium cellulosum So ce 880 = DSM 11252 in an aqueous medium containing carbon and nitrogen sources and mineral salts, aerobically cultivated in the presence of an adsorber resin,

(b) the adsorber resin is optionally separated from the culture medium with the cells and extracted with methanol and then with acetone,

(c) the combined extracts are concentrated until an aqueous phase occurs,

(d) the aqueous phase is extracted with ethyl acetate, dried and the solvent is evaporated off,

(e) the residue obtained is dissolved in methanol, the solution obtained is cooled and the precipitate which forms is separated off,

(f) subjecting the remaining solution to gel chromatography with methanol as the eluent,

(g) the fractions containing carboxylic acids (maracenins) are chromatographed on HD-SIL-18-20-60 with methanol / ammonium acetate buffer as the eluent and a fraction with w-unsaturated and receives a fraction with w-saturated carboxylic acids (maracenines),

(h) subjecting the fractions obtained to preparative HPLC chromatography and obtaining fractions with separated carboxylic acids (maracenins) and isolating the separated carboxylic acids.

The invention further relates to a process for the production of unsaturated aliphatic carboxylic acids (maracenins), characterized in that

(a) Sorangium cellulosum So ce 1128 = DSM 11253 in an aqueous medium containing carbon and nitrogen sources and mineral salts, aerobically cultivated in the presence of an adsorber resin,

(b) the adsorber resin is optionally separated from the culture medium with the cells and extracted with methanol and then with acetone,

(c) the combined extracts are concentrated until an aqueous phase occurs,

(d) the aqueous phase is extracted with ethyl acetate, dried and the solvent is evaporated off,

(e) the residue obtained is dissolved in methanol, the solution obtained is cooled and the precipitate which forms is separated off,

(f) subjecting the remaining solution to gel chromatography with methanol as the eluent,

(g) the fractions containing carboxylic acids (maracenins) on HD-SIL-18-20-60 with methanol / ammonium acetate buffer as running agent chromatographed tel and obtained a fraction with w-unsaturated and a fraction with w-saturated carboxylic acids (maracenins),

(h) the fractions obtained are subjected to preparative HPLC chromatography and fractions with separated carboxylic acids (maracenines) are obtained and the separated carboxylic acids are isolated.

Amberlite, in particular Amberlite XAD-16, can be used as the adsorber resin in the process according to the invention.

You can also dry in step (d) with sodium sulfate.

Gel chromatography can also be carried out with Sephadex, in particular with Sephadex LH-20.

Furthermore, preparative HPLC chromatography can be carried out on a reverse phase, in particular Nucleosil, preferably Nucleosil 100-C18-7.

A further embodiment of the invention relates to an agent against bacteria, in particular mycobacteria and / or nocardia, containing an unsaturated aliphatic carboxylic acid (maracenin) according to the invention in addition to conventional auxiliaries and / or carriers.

The agent according to the invention can be provided for the treatment of tuberculosis.

The invention is explained in more detail below by examples. Example 1: Maracenins A and B

A. Description of the production strain and biological activity.

1. Origin, taxonomy, morphology

The production organism was isolated at GBF in June 1992 from a soil sample from the area around Masai Mara, Kenya. It is a strain of the Myxobacterium Sorangium cellulosum (Imshenetski and. Solntseva, 1936) (= Polyangium cellulosum). The strain was designated So ce880.

The vegetative cells are cylindrical rods with round ends, usually around 1 μm thick and 3 - 6 μm long. They appear dark in the phase contrast microscope. They move smoothly. On some nutrient media, the organism forms fruiting bodies in abundance. B. on filter paper over mineral salt agar. The fruiting bodies are dark red-brown cushions and consist of a more or less large number of sporangioles, spherical or polyhedral structures with a fixed wall, 20 to 30 μm in diameter, by flattening. In the sporangioles there are myxospores, rod-shaped permanent cells of a similar shape and size to the vegetative cells, but highly refractive and drying-resistant in the phase contrast microscope.

2. Culture

The organism grows on peptone agar, e.g. B. CY agar (Casitone, Difco, 0.3%; CaCl • H2O 0.1%; yeast extract, Difco, 0.1%; agar 1.5%; pH 7.2), however a carbohydrate, e.g. B. glucose or starch must be added. The Carbohydrate can be used in a concentration of e.g. B. 0.1% can be added.

Good growth also occurs on yeast agar, e.g. B. VY / 2 agar (baker's yeast 0.5%, based on fresh weight; CaCl2 • 2H2O 0.1%; agar 1.5%; pH 7.2). The strain quickly and vigorously breaks down chitin and starch. So ce880 grows in liquid media partly clumped, partly in a homogeneous cell suspension, both in shake flasks (at e.g. 160 rpm) and in bioreactors (tested up to 100 liters). The cultivation takes place at 30 ° C under aerobic conditions.

For culture in liquid medium z. B. Medium No. 1: MD1 lm (peptone from casein, tryptically digested, Merck, 0.3%; CaCl ₂ .2H ₂ 0 0.05%; MgS0 ₄ .7H ₂ 0 0.2%; pH 7.2 ), which is supplemented by a carbohydrate source, e.g. B. glucose, starch, cellulose, each 0.1%.

Or Medium # 2: Glucose. H2O 0.5%; Peptone from casein, digested tryptically, 0.1%; MgS0 ₄ . 7H ₂ 0 0.15%; CaCl ₂ . 2H ₂ 0 0.1%; KNO3 ° ' ^2% /' sodium iron III - EDTA 8 mg / 1; K ₂ HP0 ₄ 0.0125%; Tris. HCl 0.2%; pH before autoclaving 7.4.

Or Medium No. 3: starch, soluble, 0.8%; Glucose. H ₂ 0 0.2%; Yeast extract (Difco) 0.2%; Soy flour, defatted, 0.2%; CaCl ₂ • 2H ₂ 0 0.1%; MgS0 ₄ . 7H ₂ 0 0.1%; Na-Fe III - EDTA 8 mg / 1; HEPES buffer 50 mM; pH 7.4 before autoclaving. fermentation

For example:

Bioreactor with 100 1 content, from Chemap (now Braun solutions), blade stirring system. Medium No. 3 (as above), additionally adsorber resin Amberlite XAD-16, 1%, v / v (Rohm and Haas, Darmstadt) and anti-foaming agent Tegosipon, 10 ml (Goldschmidt AG, Essen), but without HEPES.

The pH before autoclaving: 7.8; Temperature: 30 ° C, stirring speed: 100 U / min, ventilation: 1800 1 air per hour. After autoclaving, the pH was adjusted to 7.9 with sterile 5% KOH if necessary.

The fermentor was inoculated with 4.8 l of preculture, which were grown in 6 x 2 l shake flasks with 800 ml of medium 3.

During the fermentation, a drop in pH below 6.9 was prevented by adding 5% KOH.

Table 1: Course of the fermentation

Time pH p02 glucose * changes

(Hours) (%) (%)

0 (start) 7.9 100

96 7.6 75 0.7

120 _ ** 55 -

145.5 - 48 -

167.5 7.3 45 0.25

192.5 - 45 - stirring speed: 110 rpm

239.5 7.2 53 0.25

263.5 - 65 - stirring speed: 100 rpm

287.5 - 44 0.25

335.5 (harvest) 7.2 52 0.05

*) Glucose measurement with Diabur-Test 5000 from Boehringer Mannheim **) - not measured

4. Services:

The strain forms the maracenins A and B during the fermentation. Maracenin A was dissolved in methanol and applied to test sheets (6 mm in diameter) in amounts of 2 μg. These test sheets were placed on agar plates in which various test organisms were seeded in low cell density. These test plates were incubated at 30 ° C. After the test organisms had grown, the inhibitory zones were read off. The result is shown in the following table:

Table 2: Growth of test organisms

Test organism inhibitory diameter (mm)

Maracenin A

Staphylococcus aureus 0

Bacillus subtilis 0

Micrococcus luteus 0

Nocardia corallina 29

Mycobacterium phlei 48

Mycobacterium lacticola 31

Streptococcus faecalis 0

Escheri chia coli 0

Salmonella typhimurium 0

Candida albicans 0

Rhodotorula glu tinis 0

Saccharomyces cerevisiae 0

Mucor hiemalis 0

Aspergillus niger 0

Botrytis cinerea 0 To determine the minimum inhibitory concentration, maracenin A and B in various concentrations were placed in test tubes containing a suspension of the test organisms in nutrient medium. The initial cell density was 10 cells / ml. The cultures were incubated with shaking at 30 ° C for 18 to 40 hours.

The following media were used:

For bacteria: peptone from casein, tryptically digested, Merck, 0.5%; Proteose Peptone, Difco, 0.5%; Meat extract, Oxoid, 0.1%; pH 7.0.

For yeast and fungi "Mycophil" medium: Phytone Pepton, BBL, 1%; Glucose. H ₂ 0 1%. The result is shown in the following table:

Table 3: Minimum inhibitory concentration

Test organism Minimum inhibitory concentration (μg / ml]

Maracenin A B

Staphylococcus aureus 40 20

Bacill us subtle is 80 40

Corynebacterium mediolanum 20 10

Micrococcus luteus 40 40 Nocardia corallina 2.5 2.5

Mycobacterium phl ei 0.15 0.075

Mycobac terium lacti cola 0.3 0.15

The minimum inhibitory concentration for L 929 mouse fibroblasts in cell cultures was 24 μg / ml for maracenin A and B. insulation

3.5 kg of a mixture of Amberlite XAD-16 and cells were separated from a 100 l culture of Sorangium cellulosum, strain So ce880, by sieving. The batch was extracted in succession using a total of 12 l of methanol and 11 l of acetone. The combined extracts were concentrated in vacuo at a bath temperature of 35 ° C. until the water phase appeared. This was extracted three times with ethyl acetate, the combined extracts were dried over sodium sulfate and the solvent was distilled off in vacuo at a bath temperature of 35 ° C. The residue (53 g) was dissolved in 150 ml of methanol and placed in the cold (4 ° C.). The precipitate (32 g) was separated by centrifugation. The crude product remaining in solution (21 g) was fractionated by gel chromatography on Sephadex LH-20 with methanol as the eluent. The fractions containing maracenins (1.4 g) were chromatographed on HD-SIL-18-20-60 (column: 929 x 90 mm, eluent: methanol / 0.05 mM ammonium acetate buffer 80/20, flow: 7 ml / min) - phated. 132 mg of a mixture of maracenin Aτ_ and A ₂ in a ratio of 8: 1 and 119 mg of a mixture consisting of maracenin B _] _, B2 and B3 in a ratio of 25: 5: 1 were obtained. The components were separated by preparative HPLC on Nucleosil 100-C18-7 (column: 250 x 16 mm, flow: 10 ml / min) with a mobile phase mixture of methanol and 0.05 mM ammonium acetate buffer in a ratio of 70/30.

Maracenin A _x delta ' ⁵ -trans, delta ^{11, 12} -trans, delta ^14.15 - trans, delta ¹⁷ ' ¹⁸ -cis

Maracenin A ₂ delta ⁴ ' ⁵ -trans, delta ^{11, 12} -trans, delta ^14.15 - ice, delta ^{17, 18} -cis Maracenin Bi 19, 20-dihydro, delta ^{4 '5} trans, delta ^{11, 12} - trans, delta ^1/15 trans, delta' -eis

Maracenin B '19, 20-dihydro, delta ⁴ ' ⁵ -trans, delta ^{11 '12} - trans, delta ¹⁵ -cis, delta' -eis

Maracenin B- 19,20-dihydro, delta ^{4 '5} -cis, delta ^{11' 12} - trans, delta ^{14 '15} trans, delta ^{17' 18} cis

C. Physical properties

Maracenin A ^:

Molecular formula C_gH2 ₄ θ3 MG 300.40

PC: aluminum foils with silica gel 60 F ₂ 5 ₄ , layer thickness 0.2 mm, Merck art. 5554; Detection with vanillin / sulfuric acid spray reagent and heating to 120 ° C (-> violet red staining) mobile solvent: dichloromethane / acetone / methanol 85/10/5 Rf: 0.53

HPLC: column: 2 x 125 mm, material: Nucleoesil ^R 120-5C ₁₈ ; Flow: 0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV detection: 220 nm Rt_: 7.2 min

^X H-NMR (CDC1 ₃ , 300 MHz): ε. Table 4

13 C-NMR (CDCI3, 75.5 MHz): s. Table 4

IR (KBr): V 3014 (w), 2925 (m), 2855 (w), 2280 (m), 1712 (s),

1673 (m), 1662 (m), 1554 (w), 1434 (m), 1412 (m),

1351 (w), 1222 (s), 1175 (w), 1059 (m), 998 (m), 968 (m) 920 cm ^"1 (m). UV (methanol): lambda _max (log epsilon) = 208 (4.41), 227 nm

(4.47).

HPLC-MS (ESI): m / z = 299 [M-Hj

Maracenin A ₂ :

Characteristic data deviating from maracenin A _j _:

HPLC: column: 2 x 125 mm, material: Nucleosil ^R 120-5C ₁ s; Flow: 0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV detection: 220 nm R: 7.0 min

^ H NMR (CDC1 ₃ , 300 MHz): δ = 2.96 (m, 2H, 15-H), 2.82 ppm (m,

2H, 12-H

¹³ C-NMR (CDCI3, 75.5 MHz): δ = 26.1 (C-15), 25.8 ppm (C-12)

Maracenin B ^:

Molecular formula C ₁₉ H 6θ3 MG 302.41

PC: aluminum foils with silica gel 60 F25 ₄ , layer thickness 0.2 mm, Merck art. 5554; Detection with vanillin / sulfuric acid spray reagent and heating to 120 ° C (-> violet red staining) mobile solvent: dichloromethane / acetone / methanol 85/10/5 Rf: 0.51 HPLC: column: 2 x 125 mm, material: Nucleosil ^R 120-5C ₁₈ ; Flow: 0.3 ml / min; Eluent: methanol / O .01 mM ammonium acetate buffer 75/25, UV detection: 220 nm Rt: 9.9 min

^ H NMR (CDC1 ₃ , 300 MHz): s. Table 5 ¹³ C-NMR (CDCI3, 75.5 MHz): s. Table 5

IR (KBr): v = 3012 (m), 2964 (m), 2931 (m), 2875 (w), 2279 (m),

1712 (s), 1674 (m), 1662 (m), 1437 (m), 1412 (w),

1304 (w), 1288 (m), 1223 (s), 1061 (m), 968 (m), 921 cm (m).

UV (methanol): lambda _max (log epsilon) = 205 nm (4.37).

HPLC-MS (ESI): m / z = 301 [M-H]

Elemental analysis:

found 74.0% C 8.7% H 17.2% O calc. 75.6% C 8.7% H 15.9% O

Maracenin B ₂ :

Characteristic data deviating from maracenin Bη_:

HPLC: column: 2 x 125 mm, material: Nucleosil ^R 120-5C ₁₈ ; Flow: 0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV detection: 220 nm R _t : 8.9 min

^X H-NMR (CDCI3, 400 MHz): δ = 2.81 (m, 4H, 12- and 15-H) ¹³ C-NMR (CDCI3, 75.5 MHz): δ = 25.8 (C-12), 25.6 ppm (C-15)

HPLC-MS (ESI): m / z 301 [M-H]

Maracenin B3:

Characteristic data deviating from maracenin B _] _:

HPLC: column: 2 x 125 mm, material: Nucleosil ^R 120-5C ₁₈ ; Flow: 0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV detection: 220 nm R _t : 10.6 min

¹ H-NMR (CDCI3, 400 MHz): δ = 6.21 (d, J = 5.7 Hz, 1 H, 5-H),

4.75 ppm (m, 1H, 4-H)

HPLC-MS (ESI): m / z 301 [MH]

Table 4: NMR data of maracenin ^ in CDC1 ₃ (^ -H: 300 MHz; ¹³ C: 75.5 MHz)

# δ _c (in ppm) δ _H (ppm) M JH, _H (HZ)

1 178.0 -

2 34.1 2.43 t 7.3

3 21.8 2.31 q 7.3

4 108.7 5.57 German 12.0, 7.3

5 144.6 6.27 d 12.0

7 84.1 ...

8 44.1 ...

9 17.6 2.22 m

10 27.2 2.22 m

11 128.8 5.43 m

12 128.8 5.43 m

13 30.5 2.75 m

14 129.2 5.43 m

15 128.3 5.43 m

16 30.8 2.88 m

17 130.3 5.43 m

18 129.6 6.02 t 11.0

19 132.1 6.62 German 16.8, 11.0

20 117.4 5.19 d 16.8

5.10 d 10.2

Table 5: NMR data of maracenin B _x in CDCI3 (- "^" H: 300 MHz; ¹³ C: 75.5 MHz)

# δ _c (in ppm) δ _H (ppm) M JH, H (HZ)

1 178.0 -

2 34.1 2.44 t 7.3

3 21.9 2.31 q 7.3

4 108.7 5.57 German 12.0.7.3

5 144.6 6.27 d 12.0

7 84.1 ...

8 44.1 ...

9 17.6 2.23 m

10 27.2 2.23 m

11 129.1 5.43 m

12 128.5 5.43 m

13 30.5 2.72 m

14 128.7 5.43 m

15 129.0 5.43 m

16 30.3 2.72 m

17 126.9 5.33 m

18 132.3 5.40 m

19 20.5 2.03 pent 7.5

20 14.3 0.95 t 7.5 Example 2: Maracenins C and D

A. Description of the production strain and biological activity

1. Origin, taxonomy, morphology

The production organism was isolated at GBF in October 1994 from a soil sample from the white mountains in Crete. It is a strain of the Myxobacterium Sorangium cellulosum (Imshenetski and. Solntseva, 1936) (= Polyangium cellulosum). The strain was designated So cell28.

The vegetative cells are cylindrical rods with round ends, usually around 1 μm thick and 3 - 6 μm long. They appear dark in the phase contrast microscope. They move smoothly. On some nutrient media, the organism forms fruiting bodies in abundance. B. on filter paper over mineral salt agar. The fruiting bodies are yellow-brown cushions and consist of a more or less large number of sporangioles, spherical or by mutually flattening polyhedral structures with a solid wall, 20 to 30 microns in diameter. In the sporangioles there are myxospores, rod-shaped permanent cells of a similar shape and size to the vegetative cells, but highly refractive and drying-resistant in the phase contrast microscope.

2. Culture

The organism grows on peptone agar, e.g. B. CY agar (Casitone, Pifco, 0.3%; CaCl ₂ • 2H ₂ 0 0.1%; yeast extract, Pifco, 0.1%; agar 1.5%; pH 7.2), the however a carbohydrate, e.g. B. glucose or starch must be added. The Carbohydrate can be used in a concentration of e.g. B. 0.1% can be added.

Good growth also occurs on yeast agar, e.g. B. VY / 2 agar (baker's yeast 0.5%, based on fresh weight; CaCl2 • H2O 0.1%; agar 1.5%; pH 7.2). Chitin breaks down quickly and vigorously per strain. So cell28 grows in liquid medium partly clumped, partly in a homogeneous cell suspension, both in shake flasks (at e.g. 160 rpm) and in bioreactors (tested up to 100 liters). Pie cultivation takes place at 30 ° C under aerobic conditions.

For culture in liquid medium z. B. Medium No. 1: MP1 lm (peptone from casein, tryptically digested, Merck, 0.3%; CaCl ₂ .2H ₂ 0 0.05%; MgS0 ₄ .7H ₂ 0 0.2%; pH 7.2 ), which is supplemented by a carbohydrate source, e.g. B. glucose, starch, cellulose, each 0.1%.

Or Medium No. 2.: Glucose. H2O 0.5%; Peptone from casein, digested tryptically, 0.1%; MgS0 ₄ . 7H ₂ 0 0.15%; CaCl ₂ . 2H ₂ 0 0.1%; KNO3 0.2%; Sodium iron III - EPTA 8 mg / 1; K ₂ HP0 ₄ 0.0125%; Tris. HCl 0.2%; pH before autoclaving 7.4.

Or Medium No. 3: starch, soluble, 0.8%; Glucose. H ₂ 0 0.2%; Yeast extract (Pifco) 0.2%; Soy flour, defatted, 0.2%; CaCl ₂ • 2H ₂ 0 0.1%; MgS0 ₄ . 7H ₂ 0 0.1%; Na-Fe III - EPTA 8 mg / 1; HEPES buffer 50 mM; pH 7.4 before autoclaving. fermentation

For example:

Bioreactor with 100 1 content, Bioengineering, Wald, Switzerland, blade stirring system. Medium No. 3 (as above), additionally Amberlite XAP-16 adsorber resin, 1%, v / v (Rohm and Haas, Parmstadt) and anti-foaming agent Tegosipon, 10 ml (Goldschmidt AG, Essen), but without HEPES.

By pH before autoclaving: 7.8; Temperature: 30 ° C, stirring speed: 100 U / min, ventilation: 1800 1 air per hour. If necessary, the pH was adjusted to 7.9 after autoclaving with 5% KOH.

Inoculation was carried out by fermenter with 4.8 l of preculture, which were grown in 6 x 2 l shake flasks with 800 ml of medium 3 each. During the fermentation, a drop in pH below 6.9 was prevented by adding 5% KOH.

Table 6: Course of the fermentation

Time PH p02 glucose *

(Hours) (%) (%)

0 (start) 7.9 100 _ **

48 - 88 -

216 7.3 40 0.5

264 7.2 40 0.25

336 7.3 58 0

360 (harvest) 7.3 61 0

*) Glucose measurement with Diabur-Test 5000 from Boehringer

Mannheim **) - not measured 4. Services:

During the fermentation, maracenins C and P are formed per strain. To determine the minimum inhibitory concentrations, various concentrations of maracenins C and P were placed in test tubes containing a suspension of the test organisms in nutrient medium. The initial cell line was 10 ⁵ cells / ml. Pie cultures were incubated with shaking at 30 ° C for 18 to 40 hours.

The following media were used:

For bacteria: peptone from casein, tryptically digested, Merck, 0.5%; Proteose Peptone, Pifco, 0.5%; Meat extract, Oxoid, 0.1%; pH 7.0.

For yeast and fungi "Mycophil" medium: Phytone Pepton, BBL, 1%; Glucose. H ₂ 0 1%. The result is shown in the following table:

Table 7: Minimum inhibitory concentration

Test organism Minimum inhibitory concentration (μg / ml

Maracenin C P

Staphylococcus aureus 40 20

Bacillus subtilis 40 40

Corynebacterium mediolanum 40 10

Mi crococcus l u teus 40 40

Nocardia corallina 10 10

Mycobacterium phl ei 1.25 0.63

Mycobac teri um lacti cola 1.25 5

The minimum inhibitory concentration for L929 mouse fibroblasts in cell culture was 24 μg / ml for maracenin C and P. insulation

From a 100 1 culture of Sorangium cellulosum, strain So cell28, 3 kg of a mixture of Amberlite XAP-16 and cells were separated by sieving. The batch was extracted in succession using a total of 12 l of methanol and 11 l of acetone. The combined extracts were concentrated in vacuo at a bath temperature of 35 ° C. until the water phase appeared. Piese was extracted three times with ethyl acetate, the combined extracts were dried over sodium sulfate and the solvent was distilled off in vacuo at a bath temperature of 35 ° C. The residue (98 g) was dissolved in 375 ml of methanol and placed in the cold (4 ° C.). Precipitate (77 g) was separated by centrifugation. The crude product remaining in solution (21 g) was fractionated by gel chromatography on Sephadex LH-20 with methanol as the eluent. Fractions containing pie maracenine (1.7 g) were chromatographed on HP-SIL-18-20-60 (column: 929 × 90 mm, eluent: methanol / 0.05 mM ammonium acetate buffer 80/20, flow: 7 ml / min) . There were 800 mg of a mixture of maracenin C ^ and C2 in a ratio of 11: 1 and 250 mg of a mixture of maracenin _} _, and P2 in a ratio of 3: 1. The components were separated by preparative HPLC on Nucleosil 100-C18-7 (column: 250 x 16 mm, flow: 10 ml / min) with a mobile phase mixture of methanol and 0.05 mM ammonium acetate buffer in a ratio of 70/30.

Maracenin C ^ _ delta ^{4 '5} trans, delta ^{11' 12} trans, delta ^{14 '15} - trans, delta ^1' -eis

Maracenin C _' delta ^{4, 5} -trans, delta ^{11, 12} -trans, delta ^{14, 15} - cis, delta ^{17, 18} -cis

Maracenin Pη_ 19.20-dihydro, delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² - trans, delta ^{14 '15} -trans, delta ¹⁷ ' ¹⁸ -cis

Maracenin P2 19, 20-dihydro, delta ' ⁵ -trans, delta ¹¹ ' ¹² - trans, delta ^{14 '15} -cis, delta ¹⁷ ' ¹⁸ -cis

C. Physical properties

Maracenin C-,

Molecular formula C19H25O3CI MG 336.86

PC: aluminum foils with silica gel 60 F25 ₄ , layer thickness 0.2 mm, Merck art. 5554; Petition with vanillin / sulfuric acid spray reagent and heating to 120 ° C (-> violet red staining) mobile solvent: pichloromethane / acetone / methanol 85/10/5 (v / v / v) Rf_: 0.55

HPLC: column: 2 x 125 mm, material: Nucleosil ^R 120-5C ₁₈ ; Flow:

0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV petition: 220 nm Et: 8.1 min

^L H-NMR (CPCI3, 300 MHz): s. Table 8

13C-NMR (CPCI3, 75.5 MHz): s. Table 8 IR (KBr): v = 3012 (w), 2925 (m), 2859 (w), 1711 (s), 1654 (m), 1434 (m), 1412 (w), 1271 (m), 1215 (m ), 1154 (s), 1069 (w), 997 (m), 967 (m), 924 cm ^_1 (m).

UV (methanol): lambda _max (log epsilon) = 205 (4.25), 224 nm

(4.22).

HPLC-MS (ESI): m / z = 335 [M ( ³⁵ Cl) -H]

Maracenin C2:

Characteristic data deviating from maracenin C ^:

HPLC: column: 2 x 125 mm, material: Nucleosil ^R 120-5C ₁₈ ; Flow: 0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV detection: 220 nm R _t : 7.8 min

¹ H-NMR (CDCI3, 300 MHz): δ = 2.95 (m, 2H, 15-H), 2.81 ppm (m,

2H, 12-H)

13 C-NMR (CDCI3, 75.5 MHz): δ = 26.6 (C-15), 26.1 ppm (C-12)

Maracenin D ^:

Molecular formula C ₁₉ H27θ3Cl MG 338.88

PC: aluminum foils with silica gel 60 F25 ₄ , layer thickness 0.2 mm, Merck art. 5554; Detection with vanillin / sulfuric acid spray reagent and heating to 120 ° C (-> violet red staining) mobile solvent: dichloromethane / acetone / methanol 85/10/5 (v / v / v) Rf: 0.62 HPLC: column: 2 x 125 mm, material: Nucleosil ^R 120-5C ₁₈ ; Flow:

0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV detection: 220 nm Et = 9-8 min

^ ^• H-NMR (CDC1 ₃ , 300 MHz): s. Table 9 ¹³ C-NMR (CDC13, 75.5 MHz): s. Table 9

IR (KBr): v = 3011 (m), 2963 (m), 2929 (m), 2873 (w), 2856 (w), 1713 (m), 1654 (s), 1446 (m), 1409 (m ), 1270 (m), 1155 (s), 1069 (w), 967 (m), 924 (m) cm ^"1 .

UV (methanol): lambda _max (log epsilon) = 203 nm (4.16).

HPLC-MS (ESI): m / z = 337 [M ( ³⁵ Cl) -H]

Elemental analysis:

found 67.2% C 8.0% H 14.2% 0 10.5% C1 calc. 67.3% C 8.0% H 14.2% 0 10.5% C1

Maracenin D2:

Characteristic data deviating from Maracenin Dj_:

HPLC: column 2 x 125 mm, material: Nucleosil ^R 120-5C ₁₈ ; Flow: 0.3 ml / min; Eluent: methanol / 0.01 mM ammonium acetate buffer 75/25, UV detection: 220 nm R ^: 10.1 min

^ H NMR (CDCI3, 300 MHz): δ = 2.80 ppm (m, 4H, 12- and 15-H) ¹³ C-NMR (CDCI3, 75.5 MHz): δ = 25.7 (C-12), 25.6 ppm (C-15)

Table 8: NMR data of maracenin C _x in CDC1 ₃ (- "" H: 300 MHz; ¹³ C: 75.5 MHz)

# δ _c (in ppm) δ _H (ppm) M JH.H (HZ)

1 178.5 -

2 34.3 2.43 t 7.2

3 22.5 2.30 q 7.2

4 110.3 5.22 German 12.1, 7.2

5 143.2 6.26 d 12.1

7 141.5 ...

8 109.7 5.14 m

9 27.3 2.14 m

10 26.4 2.14 m

11 129.0 5.42 m

12 128.6 5.42 m

13 30.4 2.74 m

14 129.2 5.42 m

15 128.3 5.42 m

16 30.8 2.88 m

17 130.3 5.42 m

18 129.6 6.02 t 11.0

19 132.1 6.62 German 17.0, 11.0

20 117.4 5.19 d 17.0

5.10 d 11.0

Table 9: NMR data of maracenin D _x in CDC1 ₃ (: 300 MHz; ¹³ C: 75.5 MHz)

# δ _c (in ppm) δ _H (ppm) M JH, H (HZ)

1 178.5 -

2 34.3 2.42 t 7.3

3 22.5 2.28 q 7.3

4 110.2 5.22 German 12.3, 7.3

5 143.2 6.26 d 12.3

7 141.4 ...

8 109.7 5.15 m

9 27.3 2.14 m

10 26.4 2.14 m

11 129.1 5.41 m

12 128.5 5.41 m

13 30.4 2.73 m

14 128.8 5.41 m

15 128.9 5.41 m

16 30.3 2.73 m

17 126.9 5.33 m

18 132.3 5.38 m

19 20.5 2.03 pent 7.5

20 14.3 0.95 t 7.5

Claims

claims 1. Unsaturated aliphatic carboxylic acids (maracenins) of the general formula: 20 CH _x - CHy-CH = CH-CH2-CH = CH-CH2-CH = CH-CH ₂ -CH2-CH _z ==: CR-0-CH = CH-CH ₂ - CH ₂ -C0 ₂ H in which, - if x = 2, y = 1 and "" - a double bond, z = 0, R is absent and == a triple bond or z = 1, R is a Cl atom and == a double bond or, - if x = 3, y = 2 and - ^* a single bond, z = 0, R is absent and = ^* = a triple bond or z = 1, R is a Cl atom and = ^* = a double bond. 2. Unsaturated aliphatic carboxylic acid (maracenin A _] _) of the formula

Maracenin A, delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -trans, delta ¹⁷ ' ¹⁸ - cis and / or with the following parameters: MG 300.40 ^ H NMR according to Table 1, ¹³ C NMR according to Table 1, IR (KBr); ny = 3014 (w), 2925 (m), 2855 (w), 2280 (m), 1712 (s), 1673 (m), 1662 (m), 1554 (w), 1434 (m), 1412 (m ), 1351 (w), 1222 (s), 1175 (w), 1059 (m), 998 (m), 968 (m) and 920 (m) cm ^-1 ; and UV (methanol); lamda _max (log epsilon) = 208 (4.41) and 227 (4.47) nm. 3. Unsaturated aliphatic carboxylic acid (maracenin A2) of the formula 20 19 18 17 15 1 12 11 5 4 1 CH ₂ = CH-CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -C≡CO-CH = CH-CH ₂ CH ₂ -C00H delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -cis, delta ¹⁷ ' ¹⁸ -cis and / or with the following parameters: MG 300, 40, ¹ H NMR (CDC1 ₃ , 300 MHz); delta = 2.96 (m, 2H, 15-H) and 2.82 (m, 2H, 12-H) ppm, and ¹³ C NMR (CDCI3, 75.5 MHz); delta = 26.1 (C-15) and 25.8 (C-12) ppm. 4. Unsaturated aliphatic carboxylic acid (maracenin B] _) of the formula:

Maracenin B, 19, 20 -dihydro, delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} - trans, delta' - ice and / or with the following parameters: MG 302.41, ¹ H-NMR according to Table 2, ¹³ C-NMR according to Table 2, IR (KBr); ny = 3012 (m), 2964 (m), 2931 (m), 2875 (w), 2279 (m), 1712 (s), 1674 (m), 1662 (m), 1437 (m), 1412 (w), 1304 (w), 1288 (m), 1223 (s), 1061 (m), 968 (m) and 921 (m) cm ^-1 , and UV (methanol); lamda _max (log epsilon) = 205 (4.37) nm. 5. Unsaturated aliphatic carboxylic acid (maracenin B2) of the formula: 20 19 18 17 15 14 12 11 5 4 1 CH3-CH-2CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -C≡C-0-CH = CH-CH ₂ CH ₂ -COOH 19, 20-dihydro, delta ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -cis, delta' -eis and / or with the following parameters: MG 302, 41, ¹ H NMR (CDC1 ₃ , 400 MHz); delta = 2.81 (m, 4H, 12- and 15-H) and ¹³ C NMR (CDCI3, 75.5 MHz); delta = 25.8 (C-12) and 25.6 (C-15) ppm. 6. Unsaturated aliphatic carboxylic acid (maracenin B3) of the formula: 20 19 18 17 15 14 12 11 5 4 1 CH ₃ -CH-CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -C≡C-0-CH = CH-CH ₂ CH ₂ -COOH 19.20-dihydro, delta ⁴ ' ⁵ -cis, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -trans, delta ¹⁷ ' -eis and / or with the following parameters: MG 302.41 and ¹ H NMR (CDC1 ₃ , 400 MHz); delta = 6.21 (d, J = 5.7 Hz, 1H, 5-H) and 4.75 (m, 1H, 4-H) ppm. 7. Unsaturated aliphatic carboxylic acid of the formula (Maracenin C): 20 19 18 17 15 14 12 11 5 4 1 CH ₂ = CH-CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -CH = C-0-CH = CH-CH ₂ CH ₂ -COOH Cl delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -trans, delta ¹⁷ ' ¹⁸ - cis and / or with the following parameters: MG 336, 86, ¹ H-NMR according to Table 1, ¹³ C-NMR according to Table 1, IR (KBr); ny = 3012 (w), 2925 (m), 2859 (w), 1711 (s), 1654 (n), 1434 (m), 1412 (w), 1271 (m), 1215 (m), 1154 (s ), 1069 (w), 997 (m), 967 (m) and 924 (m) cm ^"1 , and UV (methanol); lamda _max (log epsilon) = 205 (4.25) and 224 (4.22) nm. 8. Unsaturated aliphatic carboxylic acid (maracenin C2) of the formula: 20 19 18 17 15 14 12 11 5 4 1 CH2 = CH-CH = CH-CH ₂ -CH = CH-CH ₂ -CH = CH-CH ₂ CH ₂ -CH = C-0-CH = CH-CH ₂ CH ₂ -COOH Cl delta ⁴ ' ⁵ -trans, delta ¹¹ ' ¹² -trans, delta ^{14 '15} -cis, delta ¹⁷ ' ¹⁸ -cis and / or with the following parameters: MG 336.86 and ¹ H NMR (CDCI3, 300 MHz); delta = 2.95 (m, 2H, 15-H) and 2.81 (m, 2H, 12-H) ppm, and ¹³ C NMR (CDCI3, 75.5 MHz); delta = 26.6 (C-15) and 26.1 (C-12) ppm. 9. Unsaturated aliphatic carboxylic acid (maracenin Dη_) of the formula:

19,20-dihydro, delta ^{4 '5} trans, delta ^{11' 12} trans, delta ^{14 '15} - trans, delta ^{17' 18} cis and / or with the following parameters: MG 338.88, ¹ HH-NMR according to Table 2, 13 C-NMR according to Table 2, IR (KBr); ny = 3011 (m), 2963 (m), 2929 (m), 2873 (w), 2856 (w), 1713 (m), 1654 (s), 1446 (m), 1409 (m), 1270 (m ), 1155 (s), 1069 (w), 967 (m) and 924 (m) cm ^-1 , and UV (methanol); lamda _max (log epsilon) = 203 (4.16) nm. 10. Unsaturated aliphatic carboxylic acid (maracenin D2) of the formula: 20 19 18 17 15 14 12 11 5 4 1

19,20-dihydro, delta ^{4 '5} trans, delta ^{11, 12} trans, delta ^{14' 15} -cis, delta ^{17 '18} -eis and / or with the following parameters: MG 338.88, ¹ H NMR (CDC1 ₃ , 300 MHz); delta = 2.80 (m, 4H, 12- and 15-H) ppm, and ¹³ C NMR (CDCI3, 75.5 MHz); delta = 25.7 (C-12) and 25.6 (C-15) ppm. 11. A process for the production of unsaturated aliphatic carboxylic acids (maracenins), characterized in that (a) Sorangium cellulosum So ce 880 = DSM 11252 in an aqueous medium containing carbon and nitrogen sources and mineral salts, aerobically cultivated in the presence of an adsorber resin, (b) the adsorber resin is optionally separated from the culture medium with the cells and extracted with methanol and then with acetone, (c) the combined extracts are concentrated until an aqueous phase occurs, (d) the aqueous phase is extracted with ethyl acetate, dried and the solvent is evaporated off, (e) the residue obtained is dissolved in methanol, the solution obtained is cooled and the precipitate which forms is separated off, (f) subjecting the remaining solution to gel chromatography with methanol as the eluent, (g) the fractions containing carboxylic acids (maracenins) are chromatographed on HD-SIL-18-20-60 with methanol / ammonium acetate buffer as the eluent and a fraction with w-unsaturated and a fraction with w-saturated carboxylic acids (maracenines) is obtained, (h) subjecting the fractions obtained to preparative HPLC chromatography and obtaining fractions with separated carboxylic acids (maracenins) and isolating the separated carboxylic acids. 12. A process for the production of unsaturated aliphatic carboxylic acids (maracenins), characterized in that (a) Sorangium cellulosum So ce 1128 = DSM 11253 in an aqueous medium containing carbon and nitrogen sources and mineral salts, aerobically cultivated in the presence of an adsorber resin, (b) the adsorber resin is optionally separated from the culture medium with the cells and extracted with methanol and then with acetone, (c) the combined extracts are concentrated until an aqueous phase occurs, (d) the aqueous phase is extracted with ethyl acetate, dried and the solvent is evaporated off, (e) the residue obtained is dissolved in methanol, the solution obtained is cooled and the precipitate which forms is separated off, (f) subjecting the remaining solution to gel chromatography with methanol as the eluent, (g) the fractions containing carboxylic acids (maracenins) are chromatographed on HD-SIL-18-20-60 with methanol / ammonium acetate buffer as the eluent and a fraction with w-unsaturated and a fraction with w-saturated carboxylic acids (maracenines) is obtained, (h) subjecting the fractions obtained to preparative HPLC chromatography and obtaining fractions with separated carboxylic acids (maracenins) and isolating the separated carboxylic acids. 13. The method according to claim 11 or 12, characterized in that Amberlite, in particular Amberlite XAD-16 is used as the adsorber resin. 14. The method according to any one of claims 11 to 13, characterized in that drying in step (d) with sodium sulfate. 15. The method according to any one of claims 11 to 14, characterized in that one carries out the gel chromatography with Sephadex, in particular with Sephadex LH-20. 16. The method according to any one of claims 11 to 15, characterized in that the preparative HPLC chromatography is carried out on a reverse phase, in particular Nucleosil, preferably Nucleosil 100-C18-7. 17. Agents against bacteria, in particular mycobacteria and / or nocardia, containing an unsaturated aliphatic carboxylic acid (maracenin) according to any one of claims 1 to 10 in addition to conventional auxiliaries and / or carriers. - 4α - 18. Composition according to claim 17 for tuberculosis treatment.