KR830002249B1 - New Antibiotic Active Compounds and Fermentative Preparations thereof - Google Patents

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Description

New Antibiotic Active Compounds and Fermentative Preparations thereof

1 is the nuclear magnetic resonance spectrum of 3-hydroxylipamycin S in 360 MH ₂ in CDCl ₃ solution.

The present invention is prepared by fermenting an antibiotic consisting of 3-hydroxylipamycin S of the following structural formula (I) and 3,31-dihydroxylipamycin S of the following structural formula (II) having the basic structure of rifamycin. It is about a method. Also provided are methods for the pharmaceutical preparation of one or more of these active compounds.

The subject matter of the present invention includes the use of these antibiotics and pharmaceutical preparation methods and methods of treating thermal diseases by administering a medicament of these compounds to warm-blooded animals, especially humans or domestic animals.

Compounds I-III of the present invention exhibit the following properties. 3-hydroxylipamycin S (1) is an amorphous red powder that is soluble in most organic solvents but insoluble in water and aliphatic hydrocarbons.

Table 1 shows the thin-layer chromatography results compared to rifamycin S.

TABLE 1

Rf of Compounds I-III and Rifamycin S (Thin Layer Chromatography of Silica Gel Plate 60F 254, Darmstadt Merk, Federal Republic of Germany)

3-hydroxy-rifamycin S (1) in the full-length desorption mass spectrum shows the maximum value of molecular ions at m / e = 711, which is consistent with Equation C ₃₇ H ₄₅ N ₁₃ and is the empirical formula for rifamycin S C ₃₇ H ₄₅ NO ₁₂ is only 16 units in weight in oxygen atoms.

The infrared spectrum (at CH ₂ C ₂ ) of 3-hydroxy-rifamycin S shows the typical band of lipinine as follows.

3450 (OH), 3350 (amide NH), 2950, 1740 (5-ring ketone), 1715 (acetate), 1685 (amide I), 1640 and 1615 (quinone C = 0), 1510 (amide II), 1415, 1380, 1325, 1295, 1155, 1065 cm ¹ .

The 360MH ₂ -NMR spectrum of the compound (see Figure 1) is almost identical to rifamycin S except for the signal at 7.3 ppm, whereas for rifamycin the signal at 7.3 ppm appears due to the H (3) quantum. It does not appear in the compound of this invention.

On the other hand, the compound of the present invention shows a C (3) atomic signal only at 162 ppm in the ¹³ C-NMR spectrum, which is substituted to the lower field side compared to rifamycin S and has an OH group at position 9). It is said to exist.

All these physical data clearly correspond to the above-mentioned structural formulas. Methods of structure determination, naming and numbering of rifamycin can be found in the literature by V. Prerogu and W. Oppulser (Holv, Act _a 56, 2279, 1973).

3,31-Dihydroxylipamycin S (II) is an amorphous red powder that is soluble in most organic solvents but insoluble in water and aliphatic hydrocarbons. Table 1 shows the thin layer chromatography results compared to rifamycin S.

In the full-length desorption mass spectrum, 3, 31-di-hydroxylipamycin S (II) shows a molecular ion with m / e = 727, which is consistent with empirical formula C ₃₇ H ₄₅ NO ₁₄ and 3-hydroxylipamycin S ( Experimental formula C ₃₇ H ₄₅ NO _{13 in} 1) differs only in weight by 16 units in oxygen atoms.

Infrared spectra (at CH ₂ C ₂ ) show substantially consistent with those of 3-hydroxy siripamycin S.

Compound (II) in the 360MH ₂ -NMR spectrum shows no signal at 7.3 ppm due to the phenomenon caused by both H (3) as described above compared to rifamycin S.

In addition, one signal of the secondary methyl group of the ansa ring does not appear. However, one AB system at 3.5 ppm reappears, possibly due to the primary OH group of C (31). In the double resonance test, 2.5 ppm H (20) coupling is coupled with vinyl quantum H (19) at 5.95ppm, H (21) quantum at 4 ppm and two H (31) quantums. Can be seen.

^{In the 13} C-NMR spectrum (see Table 2), C (31) appears at 62.3 ppm. However, compared to the eight signals of rifamycin S and 3-hydroxylipamycin S (I), only seven signals show only methyl groups between 8 and 22 ppm. All of these physical data are consistent with the estimated breakfast.

1-Desoxy-1-oxaripamycin S (III) is a light yellow amorphous powder that is soluble in most organic solvents and insoluble in water or aliphatic hydrocarbons. The thin layer chromatography results are shown in Table 1. 1-Desoxy-1-oxaripamycin S (III) differs from rifamycin G in that only C (16) -C (17) double bonds are present [G. Lansini's Suin published by Journal of Antib. 29 (4) 466 (1976).

In the mass spectrum, compound III shows molecular ions at me 683 (rifamycin G has M ⁺ at m / e685), which is consistent with empirical formula C ₃₆ H ₄₅ NO ₁₂ .

IR spectrum is substantially the same as that of rifamycin G.

The presence of C (16) -C (17), C (17) -C (18) and C (28) -C (29) double bonds in the 100MH ₂ -NMR spectrum (CDCl ₃ ). In the case of rifamycin G, C (3) quantum appears at 7.2 ppm.

^{The 13} C-NMR spectrum (in CD ₃ OD) also shows agreement with the estimation structure (see Table 2).

In the case of rifamycin, the C (1) atomic signal normally found at 185 ppm is absent. The signals of aromatic carbon atoms C (2), C (3) and C (9) also show a large change compared to rifamycin S.

The lipamycin derivatives I-III of the present invention exhibit good antibiotic activity corresponding to rifamycin S and in particular antimicrobial activity against all picture positive and negative bacteria. They exhibit superior antimicrobial activity than rifamycin, especially against Gram-negative bacteria such as Escherichia coli and Pseudomonas SP.

Compounds I-III of the present invention are obtained by separating them from fermentation materials, and these compounds are formed by fermentation alone or several species by culturing certain strains of Nocardia mediterranei described in detail below.

Separation is carried out according to separation methods such as centrifugation, filtration, solvent extraction and precipitation, crystals and chromatography (adsorption chromatography and partition chromatography). In a typical separation process, the fermentation material (culture broth) is randomly filtered using a filter aid (for example, diatomaceous earth) to remove mycelia, and organically mixed with water to some extent in a discontinuous process or a continuous process (for example, a countercurrent process). The solvent is extracted from the culture filtrate, and the solvent used is ethyl acetate or halogenated aliphatic hydrocarbons (for example, methylene chloride, chloroform, trichloroethylene). Prior to extraction, it is desirable to adjust the pH of the culture filtrate to 2-3 using an acid (e.g. using a mineral acid such as hydrochloric acid, sulfuric acid or phosphoric acid, or sometimes a strong organic acid such as hydroxyl or citric acid). Volatile parts (particularly solvents) are removed by evaporation of organic veins and the residue (crude extract) is used in the next process.

Chromatography (eg, to remove compounds I or III according to the invention or to carry away substances (for example metabolites, in particular other compounds with a basic lipamycin structure), or optionally to separate compounds I to III separately For example, the crude extract is further purified by column chromatography. Silica gel is particularly preferred as an adsorbent, preferably a solvent in which chloroform is converted to 1 to 20% methanol, 3-hydroxylipamycin S is eluted to 1 to 5% methanol, and then 3, 31-dihydroxy-lipa. Mycin S is eluted in 5-10% methanol.

The economic process can be repeated, if desired, optionally using other adsorbents and / or solvent systems. In particular, the efficiency of purification can be tested by thin layer chromatography under the above conditions which are advantageous for this. Biological test methods may be used alone or in combination with thin layer chromatography, for example, an appropriate microorganism (preferably a highly sensitive microorganism) may be used to test the antibiotic activity of an individual substance. Particularly preferred test microorganisms are Escherichia coli, and the method of separating by thin layer chromatography and combining bioautographicla detection has proved to be particularly good. In this case, when the germination plate inoculated with the test microorganism (for example, E. coli) is pressed onto the chromatographic plate to be developed, the active substance forms a corresponding inhibition zone.

In order to separate small amounts of 3-hydroxylipamycin S in pure form, it is very advantageous to use thin layer chromatography, in which case a thin silica gel layer (F 254 plate described above) is used for the preferred elution. Preference is given to mixtures of chloroform and methanol (4: 1 or 6: 1) or mixtures of ethyl acetate, acetone and water (72: 24: 4) or mixtures of toluene, acetone and methanol (5: 3: 2). Similar methods can be used to obtain 3, 31-dihydroxylipamycin S and 1-desoxy-1-oxaripamycin S purely using the same thin plate and elution system. Preferred zones are mechanically separated and extracted with a suitable solvent, preferably ethyl acetate, to separate into pure product. If necessary, the calibration process may be repeated using various adsorbents and / or solvent mixtures.

Fermentation containing 3-hydroxylipamycin S, 3, 31-hydroxylipamycin S and / or 1-dexy-1-oxaripamycin S obtained according to the present invention by culturing in aerobic conditions in liquid medium The substance is separated and used by the above-described method, and the strain used here is the bacterium of Nocardi Mediterrane derived from the parent strain Streptomyces meditellanai ATCC 13685, which is 3-hydroxylipamycin S, Having the ability to produce at least one of 3, 31-dihydroxy S and 1-decoxy-1-oxaripamycin S. Harmonized bacteria among these bacteria are resistant to streptomycin and autotrophic for amino acids is preferred.

As liquid influence media, an aqueous solution or suspension containing at least one carbon source and at least one nitrogen source (preferably mineral) is used. The carbon source is as follows. Carbohydrates, polysaccharides (such as starch), disaccharides (such as lactose and sucrose), monosaccharides (especially glucose) and beet molasses or sugar cane molasses, such as glycerin and cyclitol (eg mannitol) Industrial raw materials containing carbohydrates, such as.

Nitrogen sources are as follows.

Amino acids (especially naturally occurring α-amino acids), peptides and all proteins and their degradation products (e.g. peptone, trimton) and industrial raw materials containing ammonium salts and nitrates, nitrogen sources (e.g. Extracts, casein hydrolysates and yeast autolysates and yeast extracts).

Mixtures of nitrogen and carbon sources (e.g., various plant seeds) are also preferred and are soluble extracts, in the form of meal or pulp of soybeans (e.g. soybeans) and cereals (e.g. wheat or wheat corn) Corn steep liquor) or cottonseed and beef extract.

In addition to ammonium salts and nitrates, nutrient media may include inorganic salts of chlorides, carbonates, sulphates and phosphates of alkali and alkaline earth metals and also trace elements (eg magnesium, iron, zinc, manganese) and the like.

In a conventional manner, the nutrient medium is prepared and sterilized to inoculate the cultured production strain. Incubation is carried out under aerobic conditions (e.g., stationary surface culture or attached culture supplying oxygen) by stirring or shaking (e.g. shaking flask or known fermenter). Preferred temperatures for the culture are 20 ° and 35 ° C., preferably 22-30 ° C. and particularly preferably 28 ° C.

The culture is performed at pH 5.0 to 9.0, preferably 6.0 to 8.0, and particularly preferably near neutral. If cultured normally, there is no need to adjust the pH during fermentation. Under these circumstances, the maximum yield of 3-hydroxylipamycin is from 3 to 14 days, normally 7 days, and is deposited in the nutrient medium.

If the medium is relatively large, one or more precultures are performed in the liquid nutrient medium for a relatively short period (e.g. 2 to 3 days), and then in a significant amount of nutrient medium (e.g. 20 times that amount). It is effective to incubate stepwise until inoculation reaches the desired yield.

Strain of Nocadia Mediterranei producing at least one of the compounds of the group consisting of 3-hydroxylipamycin S, 3, 31-dihydroxylipamycin S and 1-desoxy-1-oxaripamycin S The production of the microorganism according to the present invention is obtained by culturing the mutant strain or cyclic strain of Nocdia Mediterranei for testing, and the parent strain is derived from Streptomyces mediterranean ATCC 13685 and 3-hydroxylipamycin as in the following example. It is to test the fermentation material obtained for the presence of S, 3, 31-dihydroxy silimamycin S and / or 1-desoxy-1-oxaripamycin S. Advantageous harmonized strains are obtained, for example, by crossover and selective gene exchange between two Nocardia mediterranean strains A and B derived from Streptomyces mediterranean ATCC 13685 as the parent strain. [Reference: T. Schupp et al. Journal of Bacteriology 121, 128-136 (1975). (The parent strain was deposited under the original classification name Streptomyces Seth Mediterrane (Margalith and Breth) under ATCC collection number 13685. As a result of further experiments, this name was later amended and renamed to Nocadia Mediterrane (Thieman et al.) ATCC 13685. Proposed original name used here).

Variant A and B alternately in a known manner by the mutagenesis of N-methyl-N'-nitro-N-nitrosoguanidine or ultraviolet light on the hyphae suspension of parent strain Streptomyces mediterranean ATCC 13685 Obtained and separated by selection depending on its specificity. Variant A produces mainly rifamycin B and is similar to the parent strain in that it is 50 times more resistant to streptomycin, while nutrient requirement for cysteine, lysine and leucine, say, three amino acids in growth. It has the necessary characteristics. Mutant strain B is distinguished because it is particularly sensitive to streptomycin and furthermore, it does not produce rifamycin B but merely produces rifamycin W, an intermediate of rifamycin synthesis. [Reference: R.J. white. Proc. Nat. Acad Sci. USA 71, 3260-3269 (1974). Steering is carried out in a manner known per se by culturing both mutants together and separating them according to their specificity by selection in a known manner as referenced in the T. Schupp et al.

The most important manipulation for the separation of the desired stem cells is a mixed culture obtained on selective minimal medium, such as culture No. 3, which co-cultures both variant strains A and B on a complete medium, such as culture of No. 2. Will be cultured. The peculiar feature of such a minimal medium is to add as simple a carbon and energy source as possible, for example, a simple sugar, which is glucose, which contains only an inorganic salt, an inorganic ammonia salt, as a nitrogen source. Auxotrophic A strains in such selective minimal media require amino acids such as leucine, cysteine and lysine, but cannot be grown because they are not shown and B strains cannot be grown because they are inhibited by 0.025 g of streptomycin per liter. Thus, after incubation, one colony was found on the medium used by gene exchange, which not only increased resistance to streptomycin from strain A, but also raised its properties so that it could grow from strain B without the amino acids of leucine, cysteine and lysine. Has been moved. Desired cultures to produce rifamycin homologues according to the invention from several groups selected by this method are isolated from the individual colonies and 3-hydroxylipamycin S, 3, 31-dihydroxylipamycin S and And / or is obtained by basis weight for the presence of 1-desoxy-1-oxaripamycin S. This quantification is mainly carried out by extracting the effluent with an organic solvent as described above, combining the combined thin layer chromatography, and bioautomatic testing of the desired antibiotic using E. coli as the test microorganism.

Particularly suitable and representative variant A is mutant nocadia mediterranean T 204 and particularly representative variant B is variant mutant nocadia mediterranean T 191 (all parent strains are derived from strain Streptomyces mediterranean ATCC 13685). These particular mutants capture the characteristics of mutants A and B as a whole. By selective separation as described above, the transfer of strains T 104 and T 191 was obtained, in particular, the cyclist Nocadia mediterranean R 21, which was collected on December 29, 1978, by the German collection of microorganisms from Göttingen Nocadia Meniterranei has been deposited as 1415. Certain Nocadia strains are registered in the microbial collections of Switzerland, 4002, Basel Ciba-Geigy Age, with T 104, T 191 and R 21.

3-hydroxylipamycin S according to the invention is produced because it produces lipamycin S compounds as the main product and 3, 31-hydroxylipamycin S and 1-decoxy-1-oxaripamycin S as essential ancillary substances. The preferred ring mutant, which has the above characteristics, has a 50-fold resistance to streptomycin and is autotrophic against amino acids compared to the parent strain ATCC 13685. In other respects, a characteristic difference from the parent strain ATCC 13685 could not be established, especially in morphology.

The invention also alone or as described above is used as an antibiotic to combat infections caused by bacteria, in particular together with or in combination with other antibiotics of the rifamycin type, 3-hydroxylipamycin S, 3, 31-dihydroxylipa as a drug and fungicide. The use of mycin S and 1-desoxy-1-oxaripamycin S is also included. When used as a medicament, the active substance described above is administered to warm-blooded animals, especially humans, which is preferably in pharmaceutical preparations, such as at least one conventional pharmaceutical carrier or adjuvant.

For the purpose of producing pharmaceutical preparations, each one of the compounds according to the invention, in particular 3-hydroxylipamycin S, may be mixed with inorganic or organic carrier substances suitable for external, oral or parenteral administration. Suitable materials for the carrier are those such as gelatin, lactose, starch, magnesium stearate, vegetable oil, benzyl alcohol or other medicinal carriers which do not react with the novel compounds. The pharmaceutical preparations may be present in tablets, dredges, powders, suppositories or in liquid form such as solvents, suspensions, emulsions, creams or ointments. They optionally contain adjuvants such as bactericides and / or preservatives, stabilizers, wetting agents and emulsifiers. They may also contain other useful therapeutic substances. Fungicides may also be mixed with a suitable carrier, as is known.

Dosages of the active substance (3-hydroxylipamycin S, 3, 31-dihydroxylipamycin S and 1-desoxy-1-oxaripamycin S) are administered with known antibiotics of the lipamycin type, in particular lipamycin S. Although used similarly to the principles of dose, the first depends on the type of subject, the weight, the age and the individual condition of the warm-blooded animal, the second method of administration and in particular the specific sensitivity of the causative microorganism.

The invention also relates to a compound I-III according to the invention, characterized in that the medium or the microorganisms infected by such microorganisms are treated with an antimicrobial active dosage of one of the compounds I-III of the invention, particularly 3-hydroxylipamycin S. It also relates to a method of arresting or killing at least one sensitive microbial growth of III.

The term "antimicrobial active dosage" means the amount of active substance sufficient to effectively block the particular microorganism to be treated.

The following examples illustrate the invention but do not limit its scope. The temperature was in Celsius units. The composition of the solvent mixture was given by volume ratio.

The following nutrient medium was used.

The above was filled with distilled water to make 1000 ml, the pH was adjusted to 7.0, and sterilization was performed at 120 ° C. for 20 minutes.

The above was filled with distilled water to make 1000 ml, the pH was adjusted to 7.0 with KOH before sterilization, and sterilization was performed at 120 ° C. for 20 minutes.

The above was filled with distilled water to make 1000 ml, and sterilization was performed at 120 ° C. for 20 minutes.

Under sterilization conditions, still obtained as a hot solution has the following.

a) a solution of 20 glucose dissolved in 50 g of glucose with distilled water to make 100 ml and sterilized at 120 ° C. for 20 minutes, and

b) 5 ml of streptomycin solution obtained by dissolving 0.5 g of streptomycin in 100 ml of distilled water and aseptically filtering.

The above was filled with distilled water to make 1000 ml, and sterilization was performed at 120 ° C. for 20 minutes.

Example 1

Isolation of crude ring R 21

Two hybrid strains, T104 and T191, of the Nicadia mediterranean, were hybridized to a hybrid at 40 ° C. in a 40-mL No. 1 culture medium at 250 ° C. with a rotary rotary shaker per minute for 3 days. Transfer 2 ml of the culture solution of each strain to a sterile test tube and mix thoroughly. Incubation was carried out with 0.2 ml of this mixture.

이 되게 증발에 의해 농축시키고 용출 계로서 플로로포름/메탄올(4 : 1)을 사용하여 실리카겔 60 F 254플레이트(메르크)상에 박층 크로마토그 라피로 전개시킨다. 압착에 의해 이들 전개된 박층판을 대장균 발생판(바이오-오토그램)에 옮기면 저해구역(Rf : 0.45)은 3-하이드록시리파마이신 S를 함유하는 조사하에 그것의 추출물 및 3-하이드록 시리파마이신 S를 생산하게 하는 조사하에 그것의 조환주를 나타낸다. 이 방법으로 분리된 조환주 R 21은 상기한 대로 노카디아 메디테라네이 DSM1415명으로 독일 미생물수집소에 기탁되었다.Agar gradient cultures are incubated at 28 ° C. for 6 days. (Genetic exchange between two strains T104 and T191 occurs in this state). After incubation, sufficient distilled water is added to the agar slant culture of the mycelium under aseptic conditions to make a suspension by looping marks on the agar surface. This suspension is transferred to a sterile test and agitated with a sterile quartz rod 2 to 3 mm in diameter for 2 minutes. The resulting suspension from a small operation of the hyphae is washed twice by centrifugation, resuspended in distilled water and diluted 100 times with distilled water. In each case of this diluted suspension, 0.1 ml is transferred to a Petri dish containing an optional minimal medium (= No. 3 culture medium). Inoculated Petri dishes were incubated for 10 to 14 days at 28 ° C., resulting in only autotrophic stem cells resistant to streptomycin and the original mutants remaining inhibited. In order to isolate the control strains producing 3-hydroxylipamycin S grown from individual colonies, gradient 4 of the culture medium No. 4 was inoculated and incubated at 28 ° C. for 8 days. One loop type of each mycelia of this culture is inoculated into 20 ml of culture solution No. 1 in a small Ellenmeyer flask and maintained at 28 ° C. on a rotary stirrer at 250 revolutions per minute. After 7 days of culture, the culture solution is filtered through filter paper, and the obtained culture filtrate is acidified to pH 2.5 with hydrochloric acid and extracted with the same volume of methylene chloride. The extract obtained is the original amount

This was concentrated by evaporation and developed by thin layer chromatography on silica gel 60 F 254 plate (Merck) using fluoroform / methanol (4: 1) as elution system. Transfer of these developed laminates to the E. coli generating plate (bio-autogram) by compression showed that the inhibitory zone (Rf: 0.45) underwent irradiation with 3-hydroxylipamycin S and its extracts and 3-hydroxy silipama It exhibits its moiety under investigation to produce Yisin S. Crude ring R 21 isolated in this way was deposited in the German microbial collection center as DMC1415 of Nocadia Mediterranea as described above.

Example 2

Fermentation of 3-hydroxylipamycin S (I) and / or Compounds II and III

Nocadia mediaerranei strain R 21 strains are incubated on agar inclined culture (No. 4 culture medium) for 7-8 days at 28 ℃. Five Elenmeyer flasks containing 40 ml of culture No. 1 were inoculated with mycelia of the culture. The flask is held at 28 ° C. on a rotary stirrer at 250 revolutions per minute for 72 hours. After 72 hours of growth, 150 ml of inactive culture is used to inoculate 3 liters of culture medium No. 1. Fermentation is carried out for 7 days in each case 40 ml of medium in a 200 ml Erlenmeyer flask at 28 ° C. on a rotary stirrer at 250 revolutions per minute.

Example 3

Preparation and Separation of Pure Forms of [3-Hydroxy sirifamycin S (I), 3, 31-dihydroxy siripamycin S (II) and 1-decoxy-1-oxaripamycin S (III)

The fermentation material (3 L) obtained in Example 2 was filtered through diatomaceous earth and the filtrate was adjusted to pH 2.5 with 1N hydrochloric acid and extracted three times with chloroform. The aqueous phase is discarded and the organic phase is concentrated in vacuo. The resulting dark crude extract (2.02 g) is dissolved in 200 ml of methanol, 10 g of silica gel is added and the mixture is evaporated to dryness. The powdered residue is placed on top of a chromatograph column (1 cm in diameter and 40 cm in height) of 100 g of silica gel (Merck, particle size 65 to 200 nanometers). Each fraction (25 ml each) is concentrated by evaporation in a hydrograph and dried to high vacuum. Fractions were combined based on thin layer chromatography (and optionally bioautomated quantification): The main portion of 3-hydroxysripamycin S (I) is located in the fraction containing 1.5% methanol, 1-deoxy 1-Oxalipamycin S (II) and 3, 31-dihydroxy rifamycin S (III) are found in fractions containing 5-10% methanol.

For further purification (e.g. analytical purposes) crude products I, II and III are either chloroform / methanol (4: 1) or (6: 1), ethyl acetate / acetone / water (72: 24: 4) or toluene Chromatograph on a thin plate (silica gel 60F 254, supra) with a mixture of / acetone / methanol (5: 3: 2). The desired zone is mechanically cut off and extracted with ethyl acetate. The extract is washed with dilute citric acid solution, dried and concentrated by evaporation. Pure 3-hydroxy siripamycin S and 3, 31-dihydroxy siripamycin S, respectively, were obtained as amorphous red powder. Each of the compounds is readily soluble in methanol, ethanol and other lower alkanols, acetone, dimethyl sulfoxide, ethyl acetate, dioxane, tetrahydrofuran, ether, dimethylformamide, chloroform, methylene chloride and other similar chlorinated lower aliphatic hydrocarbons. And insoluble in pentane, hexane and similar fatty hydrocarbons and water. The form in thin layer chromatography, in particular the comparison with rifamycin S, is given in Table 1.

3-hydroxysripamycin S (I)

Physical data:

Total desorption mass spectrum: M ⁺ (C ₃ H ₄₅ NO ₁₃ ) at m / e 711:

UV spectrum:

a) in ethanol: λ _max (ε) 230 (29600), 260sh, 305sh, 335sh, 602 (240).

b) in 0 / 01-N HCl: λ _max (ε) 267 (14900), 305sh, 342 (4600).

c) λ _max (ε) 245sh, 308 (24500), 440 (5200) in 0.01-N NaOH.

Infrared spectra (at CH ₂ Cl ₂ ): Bands of 3450, 3350, 2950, 1740, 1715, 1685, 1640, 1615, 1415, 1380, 1325, 1295, 1155 and 1063 cm ⁻¹ :

360MH ₂ -NMR spectrum (in CDCl ₃ ): see Table 1.

¹³ C-NMR spectrum (from CDCl ₃ : see Table 2).

3,31-dihydroxy siripamycin S (II)

Physical data:

Full length adsorption scalpel spectrum: M + at m / e 727 (C ₃ H ₄₅ NO ₁₄ ):

Infrared spectrum (at CH ₂ Cl ₂ ): 3450, 3350, 2950, 1740, 1715, 1685, 1640, 1615, 1510, 1410, 1385, 1320, 1290, 1155 and 1065 cm ^-1 Bands: 360MH ₂ -NMR spectrum (From CDCl ₃ ((takes only the most important signals)

¹³ C-NMR spectrum (in CDCl ₃ ): see Table 2.

1-Desoxy-1-oxaripamycin S (III)

Physical data:

Mass spectrum: M + at m / e 683 (C ₃₆ H ₄₅ NO ₁₂ ):

IR spectrum (at CH ₂ Cl ₂ ): band of 3450, 2950, 1740, 1710, 1650, 1595, 1520, 1245, 1155, 1060 cm ⁻¹ .

100 MH ₂ -NMR spectrum (CDCl ₃ )

¹³ C-NMR spectrum (on CD ₃ OD): see Table 2.

TABLE 2

¹³ C-NMR data of lipamycin S, 3-dihydroxylipamycin S (III) and 1-disoxy-1-oxaripamycin S (III)

Claims (1)

Streptomyces as a parent strain capable of producing at least one of the antibiotic compounds 3-hydroxy siripamycin S, 3, 31-dihydroxy siripamycin S and 1-desoxy-1-oxaripamycin S Nocidia mediterranean strains derived from mediterranean ATCC 13685 are cultured under aerobic conditions in a liquid nutrient medium to separate at least one of the compounds from the fermentation material obtained. Isin S, 31-dihydroxy siripamycin S and 1-desoxy-1-oxaripamycin S.

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