JPS60123486A - Fluoroblasticidin s and its preparation - Google Patents

Abstract

NEW MATERIAL:Fluoroblasticidin S shown by the formula I or its salts. USE:A fungicide of antibiotic for agriculture, an antibiotic showing improved effect especially on blast of rice plant. PREPARATION:A bacterium belonging to the genus Streptomyces, capable of producing blasticidin S is cultivated in the presence of fluorocytocine shown by the formula II in a culture solution containing a C source, N source, inorganic salt, a very small amount of organic omponents, etc. by shaking culture or by aerated spinner sulture at 20-40 deg.C, to give a compound shown by the formula I . Properties of monoacetate type fluoroblasticidin S. Melting pont: 215-220 deg.C. Molecular formula: C19H29N8O5F. Value of elemental analysis: theoretical value (wt%) C 48.71, H 6.24, N 23.92,) 17.08, measured value (wt%) C 48.52, H 6.30, N 23.43, O 17.21, etc.

Description

DETAILED DESCRIPTION OF THE INVENTION The invention relates to a novel antibiotic, fluoroplasticidin S or salts thereof, and a method for producing them.

Plasticidin (hereinafter referred to as BS) is known as an antibiotic obtained from the fungus Stonotomyces (Japanese Patent Publication No. 35-16449). BS or its salt is widely used as a disinfectant and is widely known as an antibiotic that is particularly effective against rice imote disease. , B
It is not only used alone, but is often used in the form of salts with organic or inorganic acids, such as benzylaminobenzenesulfone 11 (4, dodecylbenzenesulfone m salt, methanesulfonic acid). salt, lauryl sulfur Cψ salt, and consideration 3, BET is the following formula ['l
It is a basic antibiotic represented by D.

The present inventor investigated the introduction of fluorine atoms into BS in order to improve the medicinal efficacy of BS, maintain its medicinal efficacy, exhibit new pharmacological effects, and examine the possibility of application as a medical drug.

The introduction of fluorine atoms in agricultural chemicals and medicines is expected to have various effects.

For example, a compound containing a fluorine atom inhibits or suppresses the enzymatic reaction by an enzyme compared to a compound without a fluorine atom, and as a result, inhibits the growth and proliferation of microorganisms and cells. It also prolongs the pharmacological action. In addition, the fluorine atom is highly polar and changes the activity of the biological receptor near the bonding site and changes the chemical stability. Furthermore, it is also considered that hydrophobization by introduction of fluorine atoms increases the fat solubility of the compound and increases its pharmacological action. Fluorinated BS for the purpose of exhibiting these effects! # It was decided that the order would be issued. The method of directly fluorinating BS with a fluorinating agent has the extremely high risk of producing various fluoride compounds and decomposing and deteriorating BEI. The method of bonding fluorine compounds greatly changes the chemical structure peculiar to BS, and there is a great possibility that its physical function will be completely changed.

-Furthermore, it is also possible to produce BS in which fluorine atoms are introduced by decomposing BF3 into denominator side chains, partially fluorinating them, and recombining them.However, problems with the above-mentioned direct fluorination remain, and the recombination was difficult #) f1
There is a risk that the 3-build process will become extremely complicated.

As a result of examining the introduction of fluorine atoms into BS, the present inventor found that
Cultivating BS-producing bacteria using fluorocytosine as a precursor,
From the culture, they collected BS containing fluorine atoms and found scales. As shown in the above formula [...],
BS has a cytosine residue. Therefore, when culturing 3-day production bacteria using fluorocytocone as a precursor, BS
It is thought that the producing bacteria take in fluorocytocone and produce BS in which the 7-77 residues are converted to fluoro-7 tosine residues. Of course, BS can be produced without requiring a precursor for BSS production, so BS with fluorine atoms introduced is usually co-produced with BS, and Be with fluorine atoms introduced is often co-produced with BS. It is obtained by separating it from a mixture with B8. Fluoro-7tocin is a known compound represented by the following formula [111], and is used as an antifungal drug.

However, there is almost no knowledge of B8-producing bacteria, such as Lactobacilli, and it is possible to culture BS-producing bacteria using fluorocytokone as a precursor.

The non-invention is fluoroplasticidin S or its salts obtained by the above-mentioned method, and their production method fI. and plasticidin S in the presence of fluorocytocone.
A method for producing fluoroplasticidin S or its salts, which involves culturing the produced fraction and searching for fluoroplasticidin S represented by the above formula [1] or its iA group from the cultured product. Chiru.

Fluoroplasticidin s (hereinafter referred to as '-FFBS) of the present invention
) is produced by a known B8 production farm in the presence of 51 Fluorocytokun moe males. , E'19 live ant cocoon is described in the above bulletin! l+! Stombutomyces griseochromogenis (S.

BS-producing strains commonly found in Stombutomyces can be used, such as Griseoahromogene B). Furthermore, it is also possible to increase the productivity of FBS or BS by using mutant strains obtained by mutating these strains.

PBS is produced by culturing BS-producing bacteria in the presence of fluorocytoclone according to the usual method of culturing Streptomyces, first separating and collecting a mixture of FBB and Be from the culture, and then separating FBB and BStl from the mixture. obtained by For example, for about 20 minutes under shaking or aeration in a culture solution containing carbon sources, nitrogen sources, inorganic salts, organic microorganisms, etc. used for culturing fluorocytosine and normal actinocytosine.
B8-producing bacteria are cultured at ~40°C, and a mixture of FBB and BS is separated and collected from the resulting culture medium using a general antibiotic collection method, and then a mixture of FBB and BS is collected using the difference in stratification affinity. After 18 days, Baft separation and purification are performed to obtain FBB. In addition, when separating and collecting FBB from the culture solution, FBB can be used at any stage.
You can also obtain this FBS tolerance by changing BS to its salt. Furthermore, FBS or its salts can be used in the form of a mixture with Be or its salts, and separation of the two may not necessarily be necessary.

The FBB (monovinegar 11ν Arashi type) of the present invention has the following properties.

(1) Melting point 215-220° (2) Molecular formula: O+e N21 N110S F'(
3) Elemental analysis value: Q HN O theoretical value (@ 4 a y 1 6.24 25.92
17. o a actual measurement @ (fishing 4a52 6.50 23.
43 1st 21 (41U V spectrum: aI NHOI λmax 282.5 nm (t
8020) aI NaQH λmax 275 nm
(+15500) 1.5) "B-NMR spectrum (in D20): a +95 (m, 2H), 2.5
b~2.65 (In, 2H), 547 (m, 2B
), i49 (m, 1) 1), 4.12 (d, IH, J-
9Hz), 4.78 (d, IH, J-9H2), 5.8
7(d,1)1. J-10HE), 6.13(d, IH
, J-1014g), 6.47 (broad e, 1H
), Z77 (d, 1H, JH-p-6ag) Hereinafter, the present invention will be explained in detail with reference to Examples, but the present invention is not limited to these Examples.

Example [Culture method] Str@ptomycθ grown in the following VIP slant culture medium
θgriseochromogenes (KOOB-
0039) was placed in a test tube containing the following medium 15- with a platinum loop and cultured with shaking at 27°C for 2 days. Next, 2 volumes of the above preculture solution were inoculated into a 500 volume Erlenmeyer flask containing 100 volumes of the following medium lI, and 4 # of rotary cultures were carried out at 27°C. -Cultivate the 5-cape 15-milk of fluorocytocone (hereinafter referred to as FO)'4#
1 flask, 3 at a time, 24 hours after incubation,
They were added at 65 hours and 48 hours, respectively.

Agar medium (Bennett's @ ground) Yeast extract 1 (
f/ as below, meat extract 1, pegtone 2, glucose 10: pH 7, lJ culture J11J I cornstarch liquor 10 (f/ as below), soluble starch 10, meat extract 10゜peptone 10: pl
(lO medium ■ body) Sucrose 100 (V / 950 ml or less), Glucose 5, (NH4)2HPO410, KOI
, 3, M gB 04・7H202, ammonium citrate 5, FeSO4-7H20Q, 04, ZnS
O4・7 H2O0,04 (13) 0aO12・2H205(f/b Om
1) (4) and (B) were each combined and adjusted to pH 7.0 after crushing.

In order to clarify the FBB produced in FC-added culture, after removing PC from the culture solution sample using the method described below, Ba
The inhibitory activity against cillus cerθus IAM-1729 was investigated. This sample contains IF B 8 and BS, so I separately created a standard @@ for the 1i total oiliness of BE3 alone, and compared this and the sample's till.
A total of 1 PBB of harmful activity was activated.

As a result, a weak activity appeared after 72 hours of culture, the activity rapidly increased after 144 hours, and the maximum activity was observed at 192 hours.

[Purification J Culture solution f 3 After adjusting the pH to 8 with NaOH, the solution was filtered to obtain a culture solution, which was adsorbed onto Diaion BP20. Diaion 1 (P20 adsorption fraction a I N
- After eluting with HOI 60 acetate and concentrating, removing acetone, passing through column chromatography (Amberlite RA 410 (OH-)), and further column chromatography (Dowex 50WX2 (H+
)). The I & colored fraction was first washed with 5% pyrimidine to remove colored impurities, and then washed with 1.2% NH4.
01 (eluted.

The 1tE,OH elution fraction is a mixture of FBEI and BS, and these fractions +7! iit dowex 50w x 8
This was done by resin chromatography using . 2X
Using a column of 20c+++, [15M
Pyrimidine "Vinegar + *, 4? 1 costal fluid (pH 4, 7)
Use i and pre-eluted fractions: B, cereu
The inhibitory activity of S. As a result, the active ++ fraction was 700
It was divided into two, ~1200- and 1600-2700-, and the former was FBS. Next, the former was purified by vapor chromatography using a developing solvent of butanol diacetic acid:water = 2:1:1 to obtain purified FBS. The FBS obtained from 1 ton of culture solution was 50 kg.

Claims (1)

[Scope of Claims] 1. Fluoroblasticidin 8 or its salts represented by the following formula. 2. Cultivating plasticidin strawberries in the presence of fluoroctozone, and collecting fluoroplasticidin S or its salts represented by the following formula [D] from the culture. Manufacturing method for Gin S or its salts.