NO164038B - MICROBIOLOGICAL FRETE FOR THE PREPARATION OF STREPTOMYCES PURPURASCENSHPL Y-11472, DSM 2658. - Google Patents

Abstract

1. Claims for the Contracting States BE, CH, DE, FR, GB, IT, LI, LU, NL, SE A Compound of the formula I see diagramm : EP0186807,P11,F2 in which R represents a sugar combination of the composition Roa-dF-Rod or Roa-Rod-Rod, where Roa represents rhodosamine, dF represents deoxyfucose and Rod represents rhodinose, and the physiologically acceptable acid addition salts thereof. 1. Claims for the contracting State AT A process for the preparation of compounds of the Formula I see diagramm : EP0186807,P11,F1 in which R represents a sugar combination of the composition Roa-dF-Rod or Roa-Rod-Rod, where Roa represents rhodosamine, dF represents deoxyfucose and Rod represents rhodinose, and the physiologically acceptable acid addition salts thereof, which comprises fermentation of the microorganism Streptomyces purpurascens DSM 2658 in a customary manner under aerobic conditions, in the presence of a nutrient medium, addition of a chemical inhibitor after a particular fermentation time, and isolation, from the culture liquid and the mycelium, of the compounds which have been formed, using organic solvents followed by chromatography in a customary manner.

Description

The invention relates to a process for the production of new kesarirodins or acid addition salts of kesarirodin denoting anthracycline compounds, characterized in that the microorganism HPL Y-11472 - DSM 2658 is fermented in the presence of chemical inhibitors.

The new compounds have an antibacterial effect against gram-positive bacteria, and also have an effect against a number of tumor types. The new compounds have the general structure shown according to formula I:

wherein H is or 0R2 and R2 is H, or a sugar combination of the following type: (A) Roa-dF-Rod (B) Roa-Rod-Rod,

where Roa means rhodosamine, dF means deoxyfucose and Rod

means rhodinose.

The structure of these sugars is shown in the following formula pictures.

Rhodosamine (Roa) Deoxyfucose (dF) Rodinose (Rod)

The two types of kesarirodin corresponding to the sugar combination (A) and (B) are designated as kesarirodin A or kesarirodin B. Accordingly, "kesarirodin" in the present invention means either kesarirodin A or B, or a mixture of both kesarirodin. Kesarirodin has a dimethylamino group in its saccharide part and can thus form acid addition salts. To those acids which by addition reaction with kesari-

rodin forms salts, belongs to e.g. hydrochloric acid, sulfuric acid and tartaric acid.

Microorganisms for the production of kesarirodins Streptomyces purpurascens (HPL Y-11472) - DSM 2658 belongs to the class Actinomycelaters, Family Streptomycetaceae and the genus Streptomyces. The microorganism is designated below as "S. purpurascens" - DSM 2658.

Anthracycline antibiotics are demonstrably produced by various Streptomyces species and are discussed in the literature. These antibiotics play an important role in medicine for controlling tumors in cancer therapy and as antibacterial drugs. In the past, various anthracycline compounds have been prepared. Of these, danomycin and adriamycin have previously been used in the clinic for carcinoma.

Rhodomycins, iso-rhodomycins and anthracycline antibiotics derived from rhodomycin are e.g. discussed in J. Med. Chemistry 20,

957-960 (1977). Aclacinomycin is described in detail in US Patent No. 3,988,315 and by Oki et al. in J. Antibiotics

28, 830 (1975) and 32, 791-812 (1979).

Cinerubins A and B are described in U.K. Patent No. 846,130, U.S. Patent No. 3,864,489 and by Keller-Schlierlein et al in "Anti-microbial Agents and Chemotherapy", page 68 (1970), Chemical Abstracts 54, 14661 ( 1960) and J. Antibiotics 28, 830 (1975).

Further summary description of anthracycline antibiotics can be seen in "Index of Antibiotics from Antinomycetes",

Hamao Umezawa, Editor-in Chief, University Park Press, State College, Pennsylvania, USA (1976) as follows:

The invention relates to a method for production

of compounds referred to below as kesarirodines,

belongs to the class of anthracyclines and has the general formula I, by fermentation of S-purpurascens HPL Y-11472-DSM 2658 at a pH value between 6.5 and 8.5, and a tem-

temperature between 24°C and 40°C under aerobic conditions in a nutrient medium that provides carbon and nitrogen as nutrients

fer containing inorganic salts and trace elements in the absence or presence of antifoam agents such as silicone oil or desmorphene, and where after 20-36 h of fermentation a chemical inhibitor 10-80 mg/l is added, and isolation of compounds

the algae from culture liquid and mycelium after 48-56 h. in the manner described below.

Carbon sources can be glucose, sucrose, starch, gly-

cerol, dextrin, fructose, molasses, oat flour, maltose, lac-

tose and galactose. Preferred carbon sources are glucose,

starch and sucrose. Nitrogen sources can be soybean-

flour, yeast flour, yeast extract, beef extract, malt extract,

agar, peptone, casein, cottonseed oil, cherry stone powder and inorganic compounds such as ammonium salts or nitrates (eg ammonium sulphate, sodium nitrate, ammonium chloride and potassium nitrate). Preferred nitrogen sources are malt extract, yeast extract, soybean meal and potassium nitrate. If desired, the nutrient can be added in the form of inorganic salts such as

sodium chloride, magnesium sulfate, potassium chloride, potassium hydrogen and dihydrogen phosphate, calcium chloride, calcium carbonate, ammonium hydrogen phosphate, sodium hydrogen and dihydrogen phosphate, magnesium phosphate, and calcium phosphate. Iron, manganese, copper, zinc and other heavy metal salts can be used as trace elements. As chemical inhibitors, potassium cyanide, potassium hexacyanoferrate (III), methylene blue, tri-phenyltetrazolium chloride, sulfanilic acid, sulfanilamide, ascorbic acid and its salts, sodium azide, iron(III) chloride, ethylenediaminotetraacetic acid, nitrofurazine, salinomycin, dithionite, rotenone, 2, 4-dinitrophenol, erythromycin and dimethylsulfoxide. Preferred inhibitors are ascorbic acid and its salts, sodium azide and ferric chloride.

The cultivation of S. purpurascens - DSM 26 58 can take place at temperatures between 24°C and 40°C and a pH value between 6.5

and 8.5. S. purpurascens DSM-2653 is cultivated: preferably under aerobic conditions at 27°C and pH value 7.0. The chemical inhibitor such as ferric chloride, sodium azide or ascorbic acid can be added after 20 - 36 hours of fermentation to achieve a final concentration between 10 and 80 mg/litre of the fermentation liquid. Preferably, after 27 hours of fermentation, sodium azide is added to achieve a final concentration of 20 mg/litre. Fermentation is stopped after 48 to 56 hours when optimal quantities of the desired compounds have been formed. Submersed fermentation is preferred as fermentation. At the start of fermentation, antifoam agents can be added to the fermenter up to a final concentration of 0.025%.

In the culture liquid thus formed, the total kesarirodin is contained both in the mycelium and in the culture filtrate.

The course of the fermentation and formation of the anthracycline compounds can be detected by extracting the combined liquid (mycelium and culture filtrate) with an organic solvent

mean and measurement of the absorption intensity at 492 nm,

and by chromatography of the extract on silicic acid plates, as well as by the antibacterial effect against Staphylococcus aureus 209 P. Chloroform, ethyl acetate, butyl acetate, butanol, toluene and benzene can be used as organic solvents. Ethyl acetate is preferred.

Kesarirodins can be produced by extraction from these culture liquids using suitable methods. One of these methods, which is shown in the attached form I, is based on extraction. Thus, kesarirodin in the culture filtrate can, for example, be recovered by extraction with a water-immiscible solvent such as ethyl acetate, after the pH value of the filtrate has been adjusted to 7.5 - 8.0.

Kesarirodin in mycelium can be isolated by treating mycelium, which has been recovered by filtration or centrifugation, with ethyl acetate, chloroform, methanol, ethanol, acetone, butanol, methyl ethyl ketone, a hydrochloric acid solution or an acetic acid solution. The preferred solvent is acetone. After the extraction, the acetone is removed and the aqueous layer is adjusted to a pH value of 7.5 - 8.0, and extracted with ethyl acetate. The culture liquid can also be subjected to the above-mentioned extraction steps without separating the mycelium.

The ethyl acetate extract of the culture filtrate and mycelium are combined or worked up separately, concentrated and then purified according to scheme II.

Form I

Isolation of the antibiotic complex of S. purpurascens DSM 2658

Form II

Isolation and purification of kesarirodin A and B

Solvent A = chloroform : methanol : acetic acid :

Aqua dest. : triethanolamine

(80 : 10 : 10 : 2 : 0.1).

Another method for producing kesarirodin from the culture liquid is based on adsorption. The kesarirodin-containing liquid material such as e.g. the culture filtrate or the extract produced by the above-mentioned extraction method is subjected to column chromatography, liquid chromatography or another method, where a suitable adsorbent is used such as activated carbon "diaion HP-20", "XAD", aluminum oxide, silica gel or "Sephadex" or "Sephadex LH- 20". The kesarirodines can be eluted with methanol or acetone from the adsorbents. The prepared eluates are concentrated to dryness, as crude kesarirodin appears as an orange-red powder. The crude kesarirodins can be purified by repeating the above-mentioned extraction and adsorption technique as often as desired. Column chromatography with the already mentioned different absorbents, countercurrent distribution or preparative thin-layer chromatography and crystallization can be used as desired. Further purification can be achieved using high-performance liquid chromatography (HPLC).

As shown in Scheme II, fraction K yields three components of which the semi-pure fraction A is further purified to give a pure compound, which is designated below as kesarirodin A. Fraction B is further purified to give a pure compound, which is designated below as kesarirodin B. The fraction ' X' is a mixture of additional anthracycline compounds which are also being further investigated. The general structure of the kesarirodines is identical to that of formula I.

The kesarirodins A and B isolated from the fermentation fluids from S. purpurascens DSM 2658 and purified in schemes I and II have the structural formulas Ia and Ib shown below: Kesarirodin antibiotics have antibacterial action against gram-positive bacteria, fungicidal action and antitumor action,

The invention will be explained in more detail with the help of some examples.

Example 1

Cultivation of S. purpurascens - DSM 26 58 for the fermentative production of the antibiotic active complex

Streptomyces purpurascens - DSM 26 58 is kept on yeast-malt agar with the following composition:

The medium is distributed in test tubes and sterilized for 20 minutes at 121°C. The test tubes are cooled in an inclined position to produce inclined agar tubes. The slanted agar tubes are inoculated with the culture and incubated for 10 - 15 days at 28°C until good growth and spore formation can be observed. A spore suspension in Aqua dest. of an inclined agar tube is used to inoculate 5 Erlenmeyer flasks to 500 ml each containing 100 ml of the inoculum cultures in the medium, or a 5 liter aspirator bottle containing 1 liter of the same inoculum culture medium.

The composition of the inoculum culture medium

The above-mentioned medium is distributed in amounts of 100 ml in Erlenmeyer flasks up to 500 ml, or in amounts of 1 liter in suction bottles up to 5 liters and sterilized for 5 minutes at 125°C. The flasks/bottles are cooled, inoculated with spore suspensions and shaken for 72 hours at 28°C at 240 revolutions on a rotary shaker with an amplitude of 3.75 cm. With the well-grown seed culture, a 15 liter glass fermenter containing 10 liters of the production medium in a concentration of 5% by volume is inoculated. The composition of the production medium

0.025% "Desmorphen" is added as an antifoam agent to the contents of the fermenter.

10 liters of the above-mentioned medium are placed in a 15 liter fermenter. The medium is sterilized using indirect and direct steam for 28 minutes at 121°C. The fermenter is cooled and inoculated with the second stage seed culture (5% by volume). The fermentation takes place at 27°C (0.51°C) with stirring at 170-180 revolutions for 48 - 56 hours. Aeration takes place with a degree of 0.6 - 0.8 wm. During the fermentation, after approx. 24 - 32 hours as indicated below the inhibitor substance. (examples 6 - 9).

The fermenter is harvested after 48 - 56 hours, when the concentration of the antibiotic-active complex amounts to 30 - 50 micrograms per ml and the culture liquid's pH value is between 5.5 and 6.0. The swing cell volume at this time is 11 - 13 ml per 100 ml.

Example 2

Cultivation of S. purpurascens - DSM 2658 for the fermentative production of the antibiotic active complex

The culture is grown under the same conditions as in example 1 with the following media:

(1) Composition of the inoculum culture medium: 7 2 hours at 28oc. The composition of the production medium

During the fermentation, as described below, after approx. 24-32 hours the chemical inhibitor (examples 6-9). The fermenter is harvested after 48-56 hours when the concentration of antibiotic active substance is 20-40 micrograms per ml, and the culture liquid's pH value is between 5.8 and 6.2. At the time of harvest, the deposited cell volume is 11-13 ml per 100 ml.

Example 3

Cultivation of S. purpurascens - DSM 2658 for the fermentative production of antibiotic complex

The culture is grown under the same conditions as in the example

1 with the following media:

The composition of the inoculum culture medium

72 hours at 28°C. The composition of the production medium

During the fermentation, after approx. 24-32 hours as discussed below the chemical inhibitor (Example 6-9). Fermentation is stopped after 48 - 52 hours.

Example 4

Cultivation of S. purpurascens - DSM 2658 for the fermentative production of the antibiotic active complex

The culture is grown under the same conditions as in example 1 with the following medium:

Strain holding agar composition:

Composition of the graft culture medium: 72 hours at 28°C.

Composition of the production medium:

During the fermentation, a chemical inhibitor is added within 24-32 hours as discussed below (examples 6-9). The fermenter is harvested after 48-52 hours, when the concentration of the antibiotic active complex amounts to 25-40 micrograms per ml, and the culture liquid's pH value fluctuates between 6.3 and 6.4. At the time of harvest, the deposited cell volume amounts to 5.0-6.0 ml per 100 ml.

Example 5

Cultivation of S. purpurascens - DSM 2658, the fermentative production of the antibiotic active complex.

The culture works in the same way as in example 1 with the following medium:

Graft culture media composition:

72 hours at 28°C.

Example 6

' Addition of chemical inhibitor:

A concentrated solution of ascorbic acid or its salts (20 mg/ml) in aqua dest is prepared. and sterilized by sterile filtration. It gives, as mentioned in example 1-5, added to the fermentation medium from 24-32 hours after starting fermentation, with a final concentration of 20-100 mg of substance per liters are achieved.

Example 7

Addition of a chemical inhibitor:

Sodium azide in concentrated (20 mg/ml) sterile aqueous solution is added to the fermentation medium as discussed in examples 1-5, approx. 24-32 hours after starting fermentation until a final concentration of 10-80 mg substance per litres.

Example 8

Addition of a chemical inhibitor:

Iron(III) chloride is dissolved to a concentration of 20 mg/ml

in water, the solution is sterilized by filtration and added to the fermentation medium mentioned in examples 1-6 approx. 24-32 hours after starting fermentation to give a final concentration of 10-100 mg substance/litre.

Example 9

Addition of a chemical inhibitor:

Addition of a sulfonamide such as sulfanilamide, sulfacetamide or sulfonamide took place by adding a sterile concentrated (20 mg/ml) aqueous solution to the fermentation medium mentioned in examples 1-5, approx. 24-32 hours after starting fermentation, as a final concentration of 40-200 mg substance per litres.

Example 10

Isolation of kesarirodin:

Centrifuge approx. 4 0 liters of the fermentation liquid for

to separate the mycelium and culture filtrate from each other. The isolation process of kesarirodin from mycelium and culture filtrate is shown in detail in Scheme I. (a) The culture filtrate is adjusted with NaHCO 3 to a pH of 7.5, and extracted twice with 20 ml of ethyl acetate, the combined acetate extracts are then concentrated in vacuo to dryness . You get approx. 1.5 g extract designated as fraction K. (b) The separated mycelium is extracted three times with 20 ml of acetone:acetate buffer (9:1) at pH 3.5, combined extracts are concentrated under vacuum to remove acetone. The remaining aqueous layer is adjusted with NaHCO 3 to a pH

value of 7.5 and extracted three times with 30 liters of ethyl acetate. The combined extracts are concentrated under vacuum to dryness. You get approx. 6.0 g of extract designated as fraction K.

Kesarirodinin-containing extract, i.e. fraction K from mycelium and culture filtrate, can be prepared together or separately. The isolation process of kesarirodin A and B is shown in the diagram

II.

(c) 7.5 g of extract (fraction K) are applied to a 4.5 x 40 cm thin layer chromatography silica gel H-(BDH) column and eluted with the following solvent mixture: Chloroform : methanol : acetic acid : aqua dest. : triethanolamine in a ratio of 80 : 10 : 2 : 0.1. With this method, you get approx. 50 mg of the semi-pure fraction A containing kesarirodin A and approx. 60 mg of the semi-pure fraction B containing kesarirodin B.

The semi-pure fractions are first washed with petroleum ether and then separately purified by preparative thin-layer chromatography on silica gel using a solvent system consisting of chloroform : methanol : acetic acid : aqua dest. : triethanolamine in a ratio of 80 : 10 : 2.0 : 0.1. You get 27 mg of the substance kesarirodin A and 32 mg of the substance kesarirodin B.

Characterization of kesarirodin A and B:

The pure substances kesarirodin A and B are analyzed respectively. using chemical analysis and spectroscopic methods.

For maximum UV absorption, the substance is used in a concentration of 10 - 30 mg/litre. Absorption spectra were recorded in areas from 200 to 800 nm.

The proton resonance spectrum (<1>H-NMR spectrum) was recorded

a HX-270 Bruker Fourier-transform Magnetic Resonance Spectrpmeter at 270 MHz. The sample concentration was 2.6-2.7 mg/0.5 ml 99.8% CDCl 3 mixed with 0.1 ml 5% Na 2 CO 3 in 99.5% D 2 O.

Mass spectrum was determined in an MS-902S AEI, mass spectrometer (FAB (Fast-Atom-Bombardment)-Positivion method).

Physico-chemical properties of kesarirodin A:

Appearance: Orange-red powder.

Chemical formula: C4o<H>53°l4<N>

Molecular weight: 77.

UV spectrum:

X max in:

(a) methanol: 203, 234, 253, 292, 464, (sh), 478, (sh), 492, 512 (sh), 526, 575 nm. (b) 0.1 N HCl-methanol: 203, 234, 253, 292, 464 (sh),

478 (sh), 492, 512 (sh).,

526 (sh), 558 (sh) nm.

(c) 0.1 N-NaOH-methanol: 20 3, 239, 298, 554, 586 nm.

<1>H-nMR spectrum (CDCl3 + 0.1 ml 5% Na2CO3 in D2<D):

7.88 (d, J=7 Hz, 1H), 7.69 (t, J=( Hz, 1H), 7.29 (d, J=

3 Hz, 1H), 5.49 (d, J = 3 Hz, 1H), 5.21 (s, 1H), 5.03 (d, J= 3 Hz, 1H), 4.85 (s, 1H ), 4.51 (q, J=6 Hz, 1H), 4.21 (q,

J=6 Hz, 1H), 4.0 - 4.1 (complex, 2H), 3.72 (s, 1H),

3.65 (s, 1H), 3.56 (s, 1H), 3.25 (d, J=20 Hzm 1H),

2.56 (d, J=20 Hz, 1H), 2.34 (bd, J=14 Hz, 1H), 2.15

(s, 6H), 2.0 - 2.2 (complex, 4H), 1.61 - 1.9 (complex 8H), 1.17 - 1.32 (overlapping doublets, 6H), 1.14

(d, J=6 Hz, 3H), 1.07 (t, J=7.5 Hz, 3H).

Solubility:

Kesarirodin A is soluble in methanol, acetone, ethyl acetate, chloroform and weak acids, slightly soluble in hexane and petroleum ether. It forms an orange-red solution in methanol/ meanwhile assumes a purple-red color in an alkaline environment.

Thin layer chromatography:

On silica gel plates with different dissolution systems, Kesarirodin A has the R^ values shown in Table I.

The above values show that kesarirodin A has structural formula Ia.

Physcal-chemical properties of kesarirodin B:

Appearance: Orange-red powder

Chemical formula: C.riHr-0.oN

J 40 53 13

Molecular weight: 755.

UV spectrum:

X max in:

(a) methanol: 203, 234, 253, 292, 464 (sh),

478 (sh), 492, 510 (sh), 526, 582 nm.

(b) 0.1 N HCl-methanol: 203, 234, 253, 292, 464 (sh), 478

(sh), 492, 510 (sh), 526 (sh),

558 (sh) nm.

(c) 0.1 N-NaOH-methanol: 203, 239, 298, 552, 584 nm.

■"■H-NMR spectrum (CDCl3 + 0.1 ml 5% Na2C<0>3 in D2O) :

7.88 (d, J=7 Hz, 1H), 7.7 (t, J=( Hz, 1H), 7.31 (d, J=8

Hz, kH), 5.52 (d, J=3 Hz, 1H), 5.21 (s, 1H), 4.95 (s,

1H), 4.83 (d, J=3 Hz, 1H), 4.4 (q, J=6 Hz, 1H), 3.98 -

4.08 (complex, 2H), 3.78 (s, 1H), 3.56 (s, 1H), 3.46

(s, 1H), 3.26 (d, J=20 Hz, 1H), 2.58 (d, J=20 Jz, 1H),

2.25 - 2.38 (complex, 1H), 2.28 (s, 6H), 1.88 - 2.2 (complex, 4H), 1.5 - 1.85 (complex, 10H), 1, 02 - 1.35 (overlapping multiplets, 12H).

Solubility:

Kesarirodin B is soluble in methanol and acetone, ethyl acetate, chloroform and weak acids, is slightly soluble in n-hexane and petroleum ether. It forms an orange-red solution in methanol, but assumes a purplish-red color in an alkaline environment.

Thin layer chromatography:

Kesarirodin B has, on silica gel plates using different dissolution systems, the R^ values indicated in Table I.

The above-mentioned results show that kesarirodin B has structural formula Ib.

Antibacterial action of kesarirodin

A bacteriostatic effect of kesarirodin was determined. The results are given as diameter (in millimetres) of the inhibition produced by the substance in the form of a 1 mg/ml solution in a hole of 6 mm diameter in agar containing the test organism, shown in the following table, Table II. Kesarirodins are also effective against Gram-positive bacteria, and can therefore be used as antibiotics to treat fungal and staphylococcal infections, diphtheria, pneumonia, etc.

Antitumor effect of kesarirodin:

The determination of the cytostatic activity of kesarirodin took place in the manner described below on mouse L 1210 leukemia cells.

(a) Proliferation test:

With this in vitro method, after incubation of the cells with different concentrations and test substances, it is possible to determine the introduced amount of a radioactively marked DNS precursor (e.g. 14C-thiamidine) in a growing cell. Untreated L 1210 cells subjected to the same procedure serve as a control. The method is briefly summarized in the following.

L 1210 cells in exponential growth phase (5 x 10 3 /ml in RPMI 1640) are incubated for 72 hours (37°C, 5% CO 2 , 95% relative humidity) in a 96-well microtiter plate with different concentrations of the test substance. As controls, cells that are only brought into contact with the fresh medium are used.

Four holes are constructed for each drug concentration and control. After 65 hours, 50 µl of 14 C thiamidine (1.5 µCi/ml) is added to mark cell DNS. After 4 hours of incubation, the cells are harvested, DMS is separated with 5% trifluoroacetic acid, and washed with water and methanol. After drying at 50°C, the scintillation number is determined with 5 ml of scintillation liquid.

The results are expressed as a quotient of the scintillation index after incubation with the test substance with this for the control substance. With the established values, a dose-effect curve is drawn, and the value ^C^g/ is determined graphically, i.e. the concentration of the test substance which inhibits the incorporation of marked thymidine compared to the controls by around 5%. The IC,.g values are compared to those of adriamycin in the following Table III. (b) Stem cell test (continuous experiment in mykagar): This method serves to demonstrate the effect of the test substances on the growth of the leukemia cells over several generations, (as the duration of a cell cycle amounts to 10-12 hours, covered during the examination period of 7 days around 14 consecutive generations).

In this test, the effect of cytotoxic substances depends

on the reduction in the number of observable colonies compared to untreated controls. The following methods are used:

500 L 1210 leukemia cells are used per plate. The experiments are carried out by continuous incubation at different concentrations of the test substance, as the substance is mixed with the upper agar layer before applying the cells to the plate. The plates are stored for 7 days in an incubator (37°C 5% C02, 95% relative humidity). Then the number of colonies formed with a diameter of more than 60 µm is counted with an inverted microscope. The results are expressed as % rate of the colonies formed on the treated plates compared to the colonies formed by untreated controls. With the determined values, a dose-effect curve is drawn from which IC,.q values are graphically determined. The IC^^ value of kesarirodin is compared in Table III with the values for adriamycin.

As stated above, the compound according to the invention has antibacterial fungicidal and cytostatic action, i.e. therapeutic action, especially against malignant tumors in animals and humans.

The compounds and acid addition salts can therefore be used as drugs for the treatment of infectious diseases such as diseases and tumors. The compounds can be administered in different ways depending on the dosage form. Normally, the compounds are administered mixed with pharmaceutically common carriers or diluents. Thus, they can e.g. administered singly or in mixture together with carriers such as maltose or lactose, or as non-toxic complexes, e.g. as deoxyribonucleic acid complex.

A typical form of administration is injection of a solution of the compounds according to the invention in distilled water or in physiological saline solution. The solutions can be injected intraperitoneally, intravenously or intraarterially.

Daily dose and unit doses can be determined from animal experiments and also from in vitro samples, so that the total dose is administered continuously or at intervals that do not exceed a predetermined range. Thus, the total dose for a treatment cycle for cancer is approx. 0.1 - 20 mg, preferably 0.5 - 10 mg/kg body weight. This dose can be administered in equivalent fractions over a period of 7 days. For the treatment of infectious and fungal diseases, 0.1 - 10 mg, preferably 0.5 - 5 mg/kg body weight, is considered as a dosage unit. As a daily dose, 1 to 3 times the dose unit can be administered.

Claims (2)

1. Process for the preparation of kesarirodin compounds which can be used for the treatment of bacterial infectious diseases, fungal and cancer diseases with the formula in which Ri means the residue OR2, R2 means the sugar combination Roa-dF-Rod or Roa-Rod-Rod, Roa means rhodosamine, dF means desoxyfucose and Rod means rhodinose, as well as their physiologically tolerable acid addition salts, characterized in that the microorganism Streptomyces purpurascens HPL Y- 11472, DSM 2658 is fermented at a temperature of 24-40°C, pH 6.5-8.5 under aerobic conditions in a nutrient medium and where, after 20-36 hours of fermentation, a chemical inhibitor 10-80 mg/l is added and the formed compounds are isolated from the culture fluid and the mycelium after 48-56 hours by extraction with organic solvents, and subsequent chromatography. 2. Method according to claim 1, characterized in that iron (III) chloride, sodium azide or ascorbic acid is used as a chemical inhibitor.