JPH02503805A - Method and microorganisms for producing Huayalihuangin - Google Patents

Abstract

(57) [Summary] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed description of the invention] Background of the method of producing faearifungin and the invention of microorganisms (1) Field to which one invention belongs This invention is a new product called faerifungin J. Standard carbonyl pentaene macrolide composition and Streptomyces (7) It is produced by a unique strain of the genus and contained in the same composition and has antimicrobial, nematicidal and The present invention relates to compounds that have insecticidal and insecticidal properties. In particular, this invention Su griseus variety Autotrophicus variety Knob (7-tomo var, 7 var, nos AT Fayrif produced by CC53668 Concerning fangin.

(2), Prior art Molecular formula Ch sHs *O+. and Cst’He. 0. . The method of the present invention has Carbonyl pentaene macrolide (c arbonyl pentaene macrolide) is described in prior literature. has been done. However, the stereoisomers according to the prior art have limited ability to inhibit microorganisms. It has significant cytotoxicity and is unstable at room temperature.

Stereoisomer “Mikochi” obtained from Streptomyces ruba Mycoticin J. Burke, R. et al. l of Investigative Dermatology23, 1.63-168 (1954), mycotisin can be used to treat yeast and It is active against M. fungi, is rapidly inactivated, and has relatively high cytotoxicity. Mikochi Syn causes hemolysis in blood agar. The molecular formula and structural formula of mycotisin have been and later Wassermann et al. (Journal of American Chemical 5ociety 89. 6 (1967) and Ch. em-ical Communications, 1634 (1970) ) has been accurately determined and detailed analytical data has been published. The structural formula is as follows That's right.

18, R=H B, R=CH3 [wherein R is H- or CH,-, referred to as rAJ and rBJ, respectively . ] The pentaene macrolide produced in the research of Wassermann et al. It is not certain whether it is the same as that by Ke et al., since there is a difference in the data. Not.

Flavofungin has been synthesized by Bekesi in Natur e It is inactive against bacteria. 5chneider etc. (Russian Jour nal(1,967)) compared related pentaene macrolide compositions. are doing. Mycotisin (m ycoticin, my-cotbicin) is a mixture of A and B. It has been shown that Flavofungin is a 9:1 mixture of A and B. It is being Antiviral activity is described. Vetlugina, L, A, etc., in Ru5sian Journal (1974), We have published more detailed analytical data, including topography analysis. Bognar et al. (Tetrahedron Let-ters 7. 471-474 (197 0)) contains the same compounds in different ratios of mycotisin and flavofungin It is concluded that it is. The ratio of A to B in flavofungin is 9:1. It is stated that. Bognar et al. (J, C, S, Perkin I 1848 (1972)) describes flavofungin and mycotisin. There is. Analytical data has been published.

Antibiotics for a summary of the properties of flavofungin and mycotisin Vol, II, Koryyoski et al, American an　5oci-ety　for　Microbiology　(19 78), and Kirk-Othmer 3.

21-47 (1978).

the purpose The object of this invention is to produce flavofungin. ) and mycoticin, which have the same molecular formula but different steric Novel pentaene polymers containing compounds with isomeric structure, stability and antimicrobial properties A chloride composition is provided. This invention also applies to certain viruses, molds and It is also an objective to provide a broad range of antimicrobial compositions that inhibit bacteria. Ru.

Furthermore, the present invention provides antimicrobial compositions obtained from specific microbial strains. Also, the purpose. For these and other purposes, please refer to the description and drawings below. This will be understood very clearly.

Brief description of the drawing Figure 1 shows the results using methanol (MeOH) or trichloromethane (CHCI). 1 is a flowchart showing a method for isolating faearifungin.

Figure 2 shows a new product of the present invention called FUNgin (RD). The X-ray diffraction pattern of a typical pentaene macrolide composition is shown. In the product, the weight ratio of A and B is approximately 1:1.

Figure 3 shows mycotisin (MYCON) according to the prior art in which the ratio of A and B is 1=1. RD; MYC) X-ray diffraction pattern is shown.

general description This invention is based on the structural formula [In the formula, R is selected from hydrogen (A) and methyl group (B), and are all trans type. ], has a negative optical rotation, and has a molar specific force of A and B ≦ X-ray diffraction for 1:1 (turns 1.5408-43, 916 Fayari exhibits 28 crystalline peaks within a d-spacing of 5 angstroms. Carbonyl pentaene macrolyte obtained from a composition called Fungin carbonyl pentaene macrolide compound It depends. The d values are shown in Table 4.

In addition, this invention has a melting point of 209-212°C with partial decomposition, and C5aHis Molecules selected from the group consisting of O+o and (47H60016 and mixtures thereof) It has the formula: Cs i Hs e O+ by cation fast atom bombardment mass spectrometry . The molecular ions of 651 and 4017 representing Ca t Ha r O +. represents The molecular ion of 665.4237 is shown, has negative optical rotation, and has a molar ratio of 1:1. The mixture is characterized in that it has an X-ray diffraction pattern as shown in Figure 2. to the carbonyl pentaene macrolide called faearifungin. It depends.

Furthermore, this invention provides a method for treating Streptomyces gu, Lyceus var. Autotrophicus. Variant knob (7 anal dredged var, 7 var, nov) ATCC53668 and a synthetic culture medium for maintaining this Streptomyces griseus. Pertains to.

Next, this invention developed a carbonyl pentaene macromolecule called phaearifungin. A method for producing loride, comprising: Streptomyces griseus var. Totorofuikasu Variant Nobu (Shinoku Dokkou - Shoku var, 7 var, no AT CC53668 filamentous fungi were grown in an aqueous growth medium containing carbon, nitrogen and inorganic sources. When grown underground, it produces a negative optical rotation pattern and an X-ray diffraction pattern as shown in Figure 2. The present invention relates to a method for producing carbonyl pentaene macrolide.

The present invention also relates to a method for inhibiting the growth of microorganisms, and the method is based on the structural formula [In the formula, R is selected from hydrogen (A) and methyl group B, and all double bonds are It is a trans type. ], has a negative optical rotation, and the molar ratio of A and B is 1=1 The X-ray diffraction pattern for Huearifungi exhibits 28 crystal peaks in Strom's d-spacing. Carbonyl pentaene macrolyte obtained from a composition called The method is characterized in that an effective amount of the compound is made to act on microorganisms.

Furthermore, this invention is called faearifungin for inhibiting the growth of microorganisms. A mixed carbonyl pentaene macrolide composition comprising: Cs5HssO+ with a melting point of 209-212°C. and Cat's.

0. . and a mixture thereof, and has a molecular formula selected from the group consisting of C! by atomic impact mass spectrometry! 651 representing IH6゜01゜, 4017 minutes The molecular ions of 665.4237 representing the child ions and C,, H,, O, are shown. , has a negative optical rotation and ITn! or in a carrier in an amount between about 1-100μ9 per gram. The present invention relates to a composition characterized by exhibiting an X-ray diffraction pattern as shown in FIG.

As used herein, the term carrier J refers to a carrier mixed with one or more active ingredients. Pharmacologically acceptable non-toxicity which together make the same ingredient suitable for administration means the substance of The term refers to the exact amount of salt to make the composition isotonic; Other pharmacologically acceptable agents such as buffers, surfactants, coloring and flavoring agents, and preservatives water and commonly used in chemical synthesis, except when the necessary ingredients are present. Contains no low molecular weight organic solvents. As suitable solid and liquid diluents and carriers is water that can be made isotonic by adding niglucose or salt, such as the following examples: Contains buffers, non-toxic organic solvents such as paraffin or vegetable oil, alcohol, glycerin. Coal, natural rocks (e.g. kaolin, alumina, talc, limestone), synthetic rocks powders (e.g. highly dispersed silica or silicates), and sugars.

For oral administration, solid or This can be done using a liquid dosage form. Dosage units for oral administration if desired Coating or embedding particulate matter in a shape, e.g. polymer or wax microencapsulation to prolong or maintain the release of ingredients. Can also be done.

Parenteral administration includes sterile solutions and suspensions injected subcutaneously, intramuscularly, or intravenously. This can be done using liquid dosage unit forms such as. These dosage unit forms may be aqueous or suspend a weighed amount of the compound in a suitable non-toxic liquid medium for injection, such as an oil-based vehicle. or can be prepared by dissolving and sterilizing suspensions or solutions. stabilizer, Preservatives and emulsifying agents may also be added.

In general, parenteral administration is preferably O,Ol-50mg/kg per day in terms of body weight. The new one is 0.1. -10 mg/kg, and oral administration is given as a daily body weight ratio. 0.1-500mg7'kg, more preferably 0.5-100mg/kH is considered to be the amount of

Streptomyces griseus var. autotrophicus var. according to the present invention Knob (i匣王士朋Var, daretenguchimori: us var, the sword is the Budapest Treaty) The American Type Culture Collection (America) an TypeCu1ture Co11ection) ATCC It has been deposited as No. 53668. This strain is referred to herein as A composition called faerifungin J is produced.

Phaearifungin, Streptomyces griseus var. autotroph Ikasu variant knob (7 suiji var, 7 var, nov) ATCC536 68 (also known as MSU 5M0O8).

This strain was isolated from a soil sample collected from a meadow ring. Bact the composition ・Peptone, glucose, Brer Rabbit-Green (registered trademark) In a medium (A-9 medium) containing 5:10:20 g of labeled molasses in distilled water 11, It was produced by culturing under the following conditions. 400 relatively small culture batches in 21 Erlenmeyer flasks (with a baffle on the bottom) containing the medium. , the same flask was placed on a 1100-20 Orp rotary shaker at 26°C for 5-7 days. It was cultured for a period of time. A relatively large culture batch was grown in 130j2 containing 100β A-9 medium. In a fermenter, aerate at a rate of 100 ff/min and stir at 1.0 Orpm. The cells were cultured at 26°C for 5 days. Streptomyces griseus variety autotro Ficus variant Nobu (Dou!Hankaku=E strong next var.

09/β or more in 1 day.

It was isolated from soil samples collected from the department. Suspend the soil in sterilized physiological saline , serial dilutions were inoculated onto various isolation media. Colonies of this strain were collected on Czapek agar. Flat plate (sucrose 20. Og, NaN0. 3. Og, K2HPO1 1,09, MgSO4” 7 H2O0.5g, MCI 0.5g, F2504 meeting 7 HzO0.01g, Bakuto agar 15.09. Distilled water I A). Microorganisms grow well at room temperature (25°C) on most laboratory media. It multiplied rapidly. YMG agar (yeast extract, malt extract, glucose, agar and distilled water Contains 4:10:4:18g per serving. ) above, yellowish-orange and abundant in the periphery Slightly wavy colonies with aerial mycelium were formed. N, Z, Am1ne-A (NZ amine-A39 in distilled water of 11) The culture appeared powdery on agar. , on nutrient agar (Difco, Detroit, Michigan, USA) The culture had a leathery appearance. Relatively old settlements are Nocardia autotrophica (No-three strokes) A typical crack appeared. during microscope observation Not only the substrate hyphae but also the aerial hyphae appeared in a straight line with perpendicular branches. spiral body, No sporangia, spore chains or endospores were seen. This microorganism is adenine, tyro It degraded hypoxanthine, hypoxanthine, xanthine, and casein. This microorganism is Adonis. Toll, cellobiose, glucose, galactose, inositol, lactose, Maltose, mandol, melibiose, a-methyl-D-glycoside, raffi Acid was produced with north, tre/10-ose, and xylose. Acid production is arabic About North, Erythritol, Meletitose, Rhamnose, and Glucitol was not observed.

The village form of AT CC53668 is Nocardia autotrophica (Noca rdia autot), but its physiological characteristics This is similar to the physiological characteristics of Streptomyces griseus. Ta. Considering these two main characteristics, this strain I was able to recognize it as a new variant of the Usu. We are Streptomyces griseus Species Autotrophicus variety Knob (wide plastic var, 7 var, no v) nomenclature was adopted.

B. Isolation, purification and chemical characterization: Isolation and purification of faearifungin , as shown in Figure 1.

Cultures are grown in broth as shown in Figure 1 and cells are then harvested. cell is treated with methanol and the broth is treated with trichloromethane. Faye Alifa Ngin crystals (X’lS) were separated twice by cooling from methanol and then The filtrate is extracted with trichloromethane. Crystals can be combined. Therefore Hue Iarifungin melts both intracellularly and extracellularly: 20 with partial degradation. Solubility: 9-212℃ Solubility: Faearifungin is almost insoluble in methanol, Quite soluble in loloform-methanol mixture, partially soluble in water, D Completely dissolves in MSO. Previously reported pentaene macrolides are is dissolved in equal amounts in methanol at the temperature tested.

Stability: Pure faearifungin is stable at room temperature and under artificial light conditions in the laboratory. In contrast, previously reported pentaene macrolides meet these conditions. It is unstable at the bottom.

2. Data about the spectrum: No base shift with addition of 5% KOH in MeOH at 262.363.5.

The UV spectrum of peracetic acid ester of faearifungin is λMaX value is 210.261,363.5° of previously studied pentaene macrolides. Acetate ester was decomposed into three peaks at a maximum of 363.5. Hueia Rifungin acetate is not degraded.

b, Mass spectrum Kotei arifungin (C3sHssO+o and C,,H6 homologous mixture with ゜O,,).

M”650. FAB, high resolution, peak matching 651.4017 (C3sHs sO+o, M”+H).

M”664. FAB, high resolution, peak matching 665.4237 (Cs7Hs +O+o, M”+H).

C, IR-spectrum: Max 3400 (○H, 1695-lactone C =0.1610-C=C, 1570-conjugate C=C)on-' d, +3C-NMR: Lactone carbo Nyl, olefinic carbon, secondary hydroxy group with carbon band, part of lactone ring Carbon added to oxygen, methyl group, methylene group, and methyne group Resonance was shown.

e,'H-NMR: Phaearifungin has 5 adjacent C=C bonds and 1 distant and C=C bonds. All double bonds are trans-type, with two secondary (2°) Contains a methyl group and contains eight secondary (2°) -OH groups due to acetylation. It was confirmed that

Faearifungin is 13, 15, 17, 19° 21.23, 25.27 o tahydroxy-31-isopropyl-14,30-dimethyl-hentriacon ter-2,4,6,8,10,28-hexene-31-olide (13,15,17 ,19,21,23,25゜27 octahydroxy-31-1sop ropyl-14,30-dim-ethyl-hentriaconta -2,4,6,8,10,28-hex-ene　-31-olide)　I” A) and 13,15,17,21°23.25.27 octahydroxy 14. 30-dimethyl-31-iso-butyl-hentriaconta-2゜4.6,8,1 0.28-hexene-31-olide (13,15,17,21,23,25,2 7octa-hydroxy-14, 30-dimethyl-31-1s o-butyl-hen-triaconta-2,4,6,8,10,28- Hexene-31-olide) CB).

The stereochemistry of the 10H and alkyl substituents on the macrolide ring is unknown. centre Eirifungin is C,,H,,0,. and C,,H,o○1° molar ratio 1 :1 mixture.

Table 1 shows faeriefungin, mycotisin ( mycoticin and flavofungin Shows a comparison of quality.

Table 1 Appearance pale yellow yellow yellow Acicular body Crystal Crystal Melting point 210-213℃ 220-222℃ Carbonization at 210℃ 310℃ (decomposes) (decomposes) molecular formula Two Streptomyces (7) strains, namely Burke et al. and Wasse et al. According to ATCC 3348 of Rmann et al. and ATCC 53668 of the present invention. A-9 medium, and Burke et al.'s medium (Journal of Invest Igative Dermatology 23, 163-168 (1954 ) The specific rotation of the composition produced in ) was measured.

[α]r values are 0.2% solution in pyridine and dioxane, 0.2% solution in MeOH. For a 15% solution. The results are shown in Table 2.

FF -70, 5-70, 5-40, 5-52, 5-34, 0-35, 3 3 mi 3 chishin - - + 63.5 + 81.7 - (Burk e etc.) MYCN-74, 5-74, 5-58, 0-58, 0-44, 66-42, 0f Lab Fungin MYC-72, 0-73, 5-50, 0-51, 5-38, 66-38, 66 ( (from literature) FF-IJ　-46,0-54,0--,-[Note] t=time 2 minutes F = Faearifungin, grown by ATCC 53668 in A-9 medium What was produced (A: B = 1) M Y C-N = AT Produced in A-9 medium by CC3348 Compound (A: B=9: 1) M Y C= AT CC3348 The compound produced in the medium of Burke et al. (mycotisin with A:B=1:1) It is thought that. ) FF-II = Produced by ATCC 53668 in the medium of Burke et al. compound Flabofungin = (A:B=9:1) AT CC3348 is the original Streptomyces ruba (7 graves) is a producer of mycotisin A and B. . The medium of Burk et al. was prepared with mycotisin A and B by AT CC3348. α12D' (C=0.48%, dioxane) = +63.5. to produce This is the original medium used for However, Streptomyces ruba ATCC3348 is no longer grown in the original Burke culture medium. Syn A and B were not produced. AT CC3348 strain was separated in A-9 medium. A port with child formula 〇gaHssO+o (90%) and CxtH6oO+o (10%) 90:10 of polyene macrolide % composition produced. This fact was confirmed by 13C-NMR and MS. vinegar Streptomyces griseus variety autotrophicus (Streptomyces ces griseusvar, 9) AT CC53668 has the molecular formula 〇 , , H&1101° and C! ,,I(,,O,.) A 50+50 (1:1) mixture of (FF) was produced. MYC (see notes to table 2) ) was different from FF (faearifungin), in which the molar ratio of A and B is almost equal. (from the literature), which clearly distinguishes FF from MYC. It was unthinkable.

Bogner et al. reported that the specific rotation of mycotisin found by Burke et al. It was confirmed that it changes with time in oxane. [α]2D in dioxane ' value changes from +63.5° to +81.7° in 9.5 minutes and reaches zero value in 15 minutes. Reached. The final optical rotation at t=■ was -18.2°.

Flavofungin does not show this effect, probably due to the high ratio of <A:B (9:) , ). Phaearifungin has a slightly different optical rotation when used in different solvents. was changed, but maintaining the same negative value obtained after t=10 minutes. Flabofungi N and mycotisin (MYC-N or MYC) differ only in the ratio of A and B. However, faearifungin (FF) has different stereoisomeric compositions. It is believed that there is.

Dioxa of octaacetylmycotisin and octaacetylflavofungin The results obtained from tests for circular dichroism (CD) in A comparison with the CD value of Gin is shown in Table 3.

Table 3 224 -2j4 363 -19.2 224 -2-04260 -5.1 9 385 -11.9 260 -3.73343 -2.51- faa riefungin 345 -1.86371 -2.18 387 -1 0.2 362 -2.04389 -2.51 366 -9.7 380 -186347 -10 ．． 2 [Note] Phaearifungin Octa-acetate or Phaearifungin Accordingly, no CD transition was observed in the 260-240 nm region.

Table 3(a) Circular dichroism (CD) of faearifungin and mycotisin-N in MeOH Faearifungin Mikochishin-Nλ 　　　Δε　　　　　　　　　　　λ　　　　　　　Δε380　 -14.6 377 -11.4374 -16.3.　　　　　　　　　　 372 372 -11.4368 372 -11.4368 -11. 8 370 −16.6362 , , , -13.5 , 365 -15.8360 -13.5 　　　　　　360　　　-15.0354　　　　　　　 JO,'l 355 , -13.4 349 -10.1 34 8 - 7.1344 -9.6 339 3.9 Mycotisin and flavov Although the CD curves of Angin's octa-acetate are similar in shape, they are shown in Table 3. There is a quantitative difference (Bognar et al., and Brow n et al, : J, C, S, Perkin I, 1848 (197 2)). Octaacetylphaearifungin is a combination of mycotisin and flavofa It produces a different CD curve than Ngin's octa-acetate. The same curve has a quantitative shape as well. There are also differences in Table 3 (a) shows phaearifungin and mycotisin-N. This shows that the two have different CD values.

These results indicate that faearifungin has significantly different stereoisomers. Indicates that it is included. The structures of these isomers were described by Wasserman et al. Urnal of Amer-ican Chemjcal 5ociety 89. 6 (1967) and Chemical Com-munica Mycotisin produced according to the method of tions, 1634 (1970)) The structure revealed by Schreiber et al. for A and B (Schreiber et al. iber, S, L, et al, :Tetrahedron Let ters28, 6001-6004 (1987), and 1bid, 28 , 6005-6008 (1987)). 5chreiber showed that there are 64 stereoisomers.

Example 4 As shown in Figures 2 and 3, fairylifungin and mycotisin (MMC- The X-ray diffraction pattern of N) was measured. As shown in Tables 4 and 5, Arifungin has 28 crystalline peaks, whereas mycotisin ( MYp-N) has 14 similar peaks. Only about 6 peaks are both common to These results demonstrate that faearifungin has significantly different stereoisomerisms. It shows that it has a body.

Table 4 Unusual X-ray diffraction pattern of faearifungin peak Specified DI file name: FUNGIN, DI raw data file name: FUNG IN, RD sample identification symbol: fungin Measurement date: December 9, 1987 Generator set conditions: 35 kV, 20mA step size, count time: 0.020 degrees, 1.00 seconds angle range ( 2θ): 2.010-59.990 degrees AI, A2 wavelength: 1.540 56.1.54435 Angstrom d-spacing width: 1.5408-4 3.9165 Use of Angstrom monochromator/Yes Maximum peak count: 19404 cts, 19404 cps28 Identify peaks.

Crystalline 28 pieces, amorphous 0 pieces List all peaks -Table 5 Summary peaks of X-ray diffraction pattern of mycotisin Specified DI file name: MYCON, DI.

Raw data file name + MMCON, RD.

Sample identification code: mycon Measurement date: December 9, 1987 Generator set conditions: 35 kV, 20mA step size, count time: 0.020 degrees, i, oo second circumferential angle range 2θ): 2.010-59.990 degrees AI, A2 wavelength: 1.540 56.1.54435 Angstrom d-spacing width: 1.5408-43 ．． Use of 9165 angstrom monochromator: Yes Maximum peak participation count: 1156 cts, 1156 cps 14 pieces Identify peaks.

Crystalline: 14, amorphous: 0.

All peaks are listed.

Table 6 compares the minimum inhibitory concentration (MIC) of faearifungin and other antifungal agents. The concentration is expressed in μy/ml.

Table 6 Fuyialifansen 5.5 3.2 3.2 3.2 (Faerie fungin) Amphotericin B 3.2 80 16 1.6 (Amphotericin B) Microorganism 2A = Candida albicans ; B = Aspergillus fumigatus (Masu to Ga! Danshō Kudo); C = Miku Rossbolm power = Su (Mi ball four tis); D = To! J: ) Huitonuru plum (Yui Namigiri rubrum).

Antifungal agents related to prior art are listed by their common names.

*This data is based on literature.

Example 6 To confirm the data from the literature, the cultures were each grown in 100 ml of liquid A. -500rr Erlenmeyer flask containing 9 medium (with baffle attached to the bottom) hair. ) was grown in The inoculated flask was shaken at 20°rpm at 26°C. I placed it on a container. pH and antimicrobial activity were monitored daily at day 144. Antibacterial activity is A concentrated suspension (200 x 10 cells/ml) of spores or yeast-like bacteria of the test species was added to 10 Emmons agar (neopeptone, glucose, Contains 10:20:18g of Bacto agar. ) and spread it evenly over the surface. Culture broth was placed on top. Test cultures were grown for 2 days at 26°C. Bacterial inhibition The clear area based on this was used as an indicator of antibacterial activity. The production of antibacterial compounds was observed after 3 days. It reached its peak on the 8th day and declined after 11 days. During this period, the medium p H was between 6-8.

Relatively large culture batches were grown in 21 flasks containing 400 mf A-9 medium. I let it happen. Culture at 26°C for 8 days (shaking speed: 1100 rp for days 1-3, 4-5 150 rpm for days 6-8 and 20 rpm for days 6-8), then remove the culture broth. After centrifugation, the cake was extracted with methanol as shown in FIG. purified substance Dissolve in DMSO and add a known amount to Candida albicans (Can-dia a Aspergillus flavus), Aspergillus flavus, and Fusari The sample was subjected to bioassay against Fusarium sp. considerable amount of nice nystatin, pimaricin, and amph Dissolve am-photericin B in DMSO, tested at the same time. Huearifungin is Aspergillus fumigatus (A, 4), it is more active than amphotericin B and slightly more active than pimaricin. It had a lower activity and showed superior activity than nice cutin. Candida albicans ( C. albicans), the compound phaearifungin is effective against amphoteri It had similar activity to SynB. Fusarium spp. Earifungin exhibited superior activity to any of the three antibiotics tested. Ta. Additionally, phaearifungin is effective against a wide range of bacterial species at low minimum inhibitory concentrations. Ru.

Table 7 shows faeriefugin against various bacteria and fungi. ngin), amphotericin B and the minimum inhibitory concentration (MIC) of nystatin, μti /m! It is shown in

Table 7 Microorganisms Ricin B Tatin As you can see from this table, phaerifungin is are active against a large number of bacterial strains, whereas compositions according to the prior art are not .

Tables 8 and 9 show flavofungin and flavofungin, respectively. This shows the activity already reported for mycoticin. .

Table 8 Burke et al, : Journal of Investig Active Dermatology, 163 (1,954) Table 9 Table 10 shows faeriefungin, mycotisin (mycoticin) and flavofungin (fla-vofungin) ) shows a summary of the biological activities of

Table 10 Antiviral effects of carbonyl-pentaene macrolides except for phaerifungin Influenza A and B virus activity was determined by M, A, 5chneider, etc. virus, the smallpox vaccine virus, and the Rous sarcoma virus. (Schneider et al,: Ru5sian Jou rnal.

1967). It was reported that flavofungin was the most active against the above-mentioned viruses. It has been tell.

Example 7 Table 11 shows the toxicity of faearifungin on human red blood cells.

Table 11 Concentration Degree Phaearifungin Amphotericin B100 μ9 /rnI! Poisonousness 20 μ9/ml Toxicity: 4μ9/ml Non-toxic: 0.8μt i/rnl Non-toxic As seen in the non-toxic is nontoxic at low concentrations in the tract.

Example 8 Streptomyces (rib! konkakuji) containing faeiarifungin ATCC536 The broth of 68 is also highly effective against larvae of the mosquito (Nedes aegyptii - Aedes7). It has been shown to have a highly toxic and rapid response (more than 50% killing within 2 hours).

In contrast, pure mycotisin (MMC-N) is only mildly toxic to mosquito larvae. (35% mortality in 24 hours at a concentration of 100 ppm), and this The other ingredients produced by ATCC 3348 are probably the main insecticidal part. It shows what will happen. Broth is Nematode Panagrelus Recipe Bath (7re It is mildly toxic (quickly → slowly) to cultures of P. divius). It was.

The biosynthesis of mycotisin A and B was carried out by Wassermann et al. studied using cetate, propionate and methionine (Chem, C communication, 1634 (1970)). The result is a macrolide ・Showed that pentaene ring is biosynthesized via acetate. alkyl side chain and two methyl group substituents, i.e. methyl substituents at C-14 and C-30 and C- The isopropyl (isobutyl) side chain at 31 is biosynthesized from propionate. (Wassermann et al.: Chem, Communica tion, 1634 (1970)). Methionine uptake has not been reported. Not yet.

In the case of faearifungin, experiments using 1IC-labeled acetate revealed that the ring carbon The uptake of acetate was shown for the element. Methyl group at C-14 and C-30 Propionate was biosynthesized. C-31 carbon and Al at C-31 For kill side chains (isopropyl or isobutyl), acetate and propionate Nor was it incorporated. These experiments showed that the production of phaerifungin was via a biosynthetic pathway different from that reported for cotisine production. It was confirmed that this process occurs and that the compounds are different for this reason.

Phaearifungin can be used to control plant or animal pathogens. pharmacology Various well-known solid or liquid carriers for medical, veterinary or phytopharmacological uses may be used. , can be used. The composition of the present invention is applied topically and is highly effective. It is also possible to

The examples described above are merely to illustrate the invention; The invention is to be limited only by the scope of the following claims.

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Claims (1)

[Claims] 1. Structural formula ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R is selected from hydrogen (A) and methyl group (B), and are all trans type. ] X-rays with a negative optical rotation and a molar ratio of A:B of 1:1 The diffraction pattern is within the d-spacing of 1.5408-43.9165 angstroms. From a composition called Huayalihuangin, which has 8 crystalline peaks. The resulting carbonyl pentaene macrolide compound. 2. Carbonyl-pentaene-macrolide group called phaearihuangin is a C36H58 compound with a melting point of 209-212°C with partial decomposition. Molecules selected from the group consisting of O10 and C37H60O10 and mixtures thereof has the formula and represents C36H50O10 by cation fast atom bombardment mass spectrometry. 665.423 representing the molecular ion of 651.4017 and C37H61O10 7 molecular ions are shown, each with negative optical rotation, in a 1:1 molar ratio mixture. A composition characterized by having an X-ray diffraction pattern shown in FIG. 3. A mixture with a molar ratio of 1:1 has a peak in the ultraviolet absorption spectrum in methanol. , at wavelengths of 262 nm and 363.5 nm. 4. Streptomyces griseus variety Autotrophicus variety knob (Streptomyces griseus var. autotrophy cus var. nov) Produced by ATCC 53668, with negative rotation A carbonyl pentaene macromolecule with a luminous intensity and an X-ray diffraction pattern shown in FIG. Loride. 5. Streptomyces griseus var. Autotrophicus var. Knob ( Striptomyces griseus var. autotrophic us var. nov) ATCC 53668, assimilable carbon, In an aqueous nutrient medium containing nitrogen and mineral sources, A biologically pure culture that produces recoverable amounts of antimicrobial substances. 6. (a). Streptomyces griseus var. Autotrophicus var. Knob (Streptomyces gri-seus vsr. autotro phicus var. nov) ATCC 53668, and (b). This space A composition comprising a synthetic growth medium for maintaining Treptomyces griseus. 7. Carbonyl pentaene macrolide called phaearihuangin A method of producing Streptomyces griseus var. autotrophy. Streptomyces griseus var.au to-trophicus var. nov) ATCC 53668 filamentous Bacteria are grown in an aqueous growth medium containing sources of carbon, nitrogen, and inorganic substances to achieve a negative optical rotation. The above carbonyl pentaene having a turn and an X-ray diffraction pattern shown in FIG. How to produce Yachloride. 8. 8. The method of claim 7, wherein the growth medium includes components obtained from blackstrap molasses. 9. Organically extract carbonyl pentene macrolide from the filamentous fungus and growth medium. Carbonyl pentaene macrolide is extracted using a solvent and then extracted from an organic solvent. 8. The method of claim 7, wherein: 10. Carbonyl pentaene macrolide was extracted from the filamentous fungus using methanol. and extracting the growth medium from the growth medium using trichloromethane, respectively. the method of. 11. The growth medium is centrifuged and the separated liquid is extracted with a solvent to extract macrolides. The cake was homogenized with methanol and then extracted with methanol. After treatment and cooling, the carbonyl pentaene macrolide was obtained and separated from methanol. 10. The method of claim 9. 12. 12. The method according to claim 11, wherein the separated liquid is extracted using trichloromethane. 13. Carbonyl pentene macrolide from methanol as yellow crystals 13. The method of claim 12 of separating. 14. carbonyl pentene macrolide by cooling the solvent. 10. The method of claim 9, further comprising depositing. 15. A method of inhibiting the growth of certain microorganisms, which is based on the structural formula ▲Contains mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R is selected from hydrogen (A) and methyl group (B), and are all trans type. ] X-rays with a negative optical rotation and a molar ratio of A:B of 1:1 The diffraction pattern is within the d-spacing of 1.5408-43.9165 angstroms. From a composition called faeifarifungin, which has 8 crystalline peaks. An effective amount of the obtained carbonyl pentaene macrolide compound was applied to microorganisms. How to make it work. 16. A method for inhibiting the growth of microorganisms, which involves partial decomposition and has a melting point of 209- 212℃, C36H58O10 and C37H60O10 and mixtures thereof It has a molecular formula selected from the group consisting of The molecular ion of 651.4017 representing C36H50O10 and C37H61 The molecular ions of 665.4237 representing O10 are shown, respectively, and have negative optical rotations. characterized in that a mixture with a molar ratio of 1:1 exhibits the X-ray diffraction pattern shown in FIG. An effective amount of a composition called faealifuangin, which is said to be effective against microorganisms, is How to make it work. 17. A method for inhibiting the growth of microorganisms, including Streptomyces griseus Variant Autotrophicus variant Knob (Streptomyces gris eus var. autotrophicus var. nov) ATCC 53668, which has a negative optical rotation and has the X-ray diffraction pattern shown in Figure 2. An effective amount of carbonyl pentaene macrolide containing How to make it work. 18. Claim 17, wherein the carbonyl pentaene macrolide is applied topically. Method. 19. A claim in which the carbonyl pentaene macrolide is mixed with a carrier in advance. 18 methods. 20. Carbonyl pentaene macrolide and carrier against microorganisms. 18. The method of claim 17, wherein the amount of reaction is about 1-100 μg per ml or 1 g. 21. The microorganism is selected from the group consisting of fungi, bacteria and viruses. 18. The method of claim 17. 22. 18. The method of claim 17, wherein the microorganism is a plant pathogen. 23. 18. The method of claim 17, wherein the microorganism is an animal pathogen. 24. A mixed composition for inhibiting the growth of microorganisms, which is a fusion composition that involves partial decomposition. point is 209-212℃, C36H58O10 and C37H60O10 and It has a molecular formula selected from the group consisting of a mixture of these, and is a cationic fast atomic bombardment material. The molecular ion of 651.4017 representing C36H50O10 and The molecular ions of 665.4237 representing C37H61O10 are shown, respectively, A mixture with a negative optical rotation and a molar ratio of 1:1 has the X-ray diffraction pattern shown in Figure 2. Carbonyl pentaene macro called huayalihuangin The ride composition is contained in the carrier in an amount of about 1-100 μg per ml or 1 g of carrier. mixed composition. 25. A carbonyl-pentaene-macrolide mixture with a molar ratio of 1:1 is converted into methanol. The peaks of the ultraviolet absorption spectrum were measured at wavelengths of 262 nm and 363.5 nm. 25. The mixed composition of claim 24.