JPH01168694A - Novel antifungal antibiotic substance dexylosylbenanomicin b and production thereof - Google Patents

Abstract

NEW MATERIAL:A compound expressed by the formula, salts or esters thereof. In addition, dexylosylbenanomicin B hydrochloride has the following physico- chemical properties. Reddish brown powder. Molecular formula; C34H34N2 O14.HCl. Mass spectrum (FD-MS); m/z 696(M+2)<+>. Melting point; 180 deg.C (decomposition). Specific rotatory power; [alpha] =+396 deg. (c=0.05, 0.05N HCl). Solubility; slightly soluble in chloroform, ethyl acetate and acetone and soluble in water, methanol, dimethyl sulfoxide and DMF. An amphoteric substance, etc. USE:An antifungal agent. PREPARATION:An antibiotic substance benanomicin B is chemically converted.

Description

Detailed Description of the Invention (Industrial Application Field) The present invention provides a novel antifungal antibiotic dexylosyl penanomycin B (D@xylosylbsnanomycin
in B) and its manufacturing method.

(Prior Art) As a compound having structural characteristics similar to the antifungal antibiotic dexylosyl penanomicin B according to the present invention, pennanomicin and B (patent application filed in 1983 filed by the present applicant) have been proposed.
-277.692), KS-619-I (Mat
guda et al.: r J, Antibloties J
4Q.

+104-1114 (19g7)), G-2N and G-2A (Gerb@r et al.: rCanad,
J, Chem, J 62.2818-2821 (1
9843E and the like are known. However, the antifungal antibiotic dexylosylpenanomycin yB+2 differs from these substances in physicochemical and biological properties and is clearly distinguishable from these substances.

(Problems that the invention seeks to resolve) Various antibiotics produced by microorganisms have been known, but not many effective anti-fungal antibiotics have been found. In the medical field of infectious diseases, there is always a demand for new antifungal antibiotics.

The present inventors have previously reported that the antibiotics benanomycin and B1 with antifungal activity and actinomycin 1iMH193-16F
He invented the production of penanomycins A and B by culturing A strain A (Feikoken Bacteria No. 9529) (Japanese Patent Application No. 62-277.692; filed on November 2, 1986). Penanomycin B was found to be a substance represented by the following formula (II).

An object of the present invention is to provide a novel antifungal antibiotic dexylocurbenanomycin B1 and a method for producing the same.

(Means for Solving the Problems) The gist of the first invention is that the novel antifungal antibiotic dexylocurbenanomycin B or its salt or ester represented by the above 2) be.

Dexylosylpenanomycin B hydrochloride has the following properties.

ζ The gist of the present invention of Saiga 2 is a method for producing an antifungal antibiotic dexylosylbenanomycin B by chemically converting the antibiotic pennanomicin B.

Here, "chemically converting" includes acid hydrolysis of benanomycin B, or treatment with a base after alcoholysis. Raw material benanomycin B
The manufacturing method is disclosed in the patent application filed by the present applicant, Kawahara, 1986-277.
Although it is described in the specification of No. 692, the manufacturing example is a reference example below! ~3.

1. Production of dexylosylbenanomycin B Benanomycin B can be used with inorganic acids such as hydrochloric acid and sulfuric acid, or with acetic acid, trifluoroacetic acid, p-toluenesulfonic acid,
When acid hydrolysis is performed with an organic acid such as benzenesulfonic acid,
Dexylosylpenanomycin B is produced in the reaction solution. Furthermore, when penanomycin B is subjected to alcoholysis, for example, methanorinsing, and then treated with a base such as sodium hydroxide or potassium hydroxide, dexylosyl penanomycin B is produced in the reaction solution.

Dexylosylpenanomycin B is collected from these reaction solutions using conventional separation methods that take advantage of its properties, such as solvent extraction, ion exchange resin method, adsorption or distribution column chromatography, and r-filtration method. , dialysis method, precipitation method, etc.
They can be extracted and purified singly or in appropriate combinations. For example, dexylosylpenanomycin B in the reaction aqueous solution is absorbed by the synthetic adsorbent Diaion HP-20 (
It has been absorbed by several Mitsubishi Motors Corporation11) and others. To further purify dexylosylbenanomycin B, adsorbents such as silica glu (Wacoal C-300, Wako Tsumugi Kogyo Co., Ltd., etc.), alumina, Sephadex LH-20 (7 Almacia Co., Ltd.), etc. It is recommended to use clotgelography.

Dexylosyl pennanomycin B produced in the reaction solution in this manner can be separated in a free form, that is, as dexylosyl pennanomycin B itself. Also,
A solution containing dexylosylpenanomycin B or a concentrated solution thereof with an acid, for example, an inorganic acid such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, or an organic acid such as acetic acid, alkylsulfonic acid, etc., or a commonly available acid such as Bases, i.e. alkali metal compounds such as sodium hydroxide, potassium hydroxide, alkaline earth metal compounds such as calcium hydroxide, magnesium hydroxide, inorganic bases such as ammonium salts, ethanolamine, triethylamine, dicyclohexyl When treated with an organic base such as an amine during each step of purification, dexylosylbenanomycin B is converted into the form of its corresponding salts and separated.

Alternatively, the salts of dexylosylpenanomycin B thus produced can be converted into the free form, dexylosylpenanomycin B itself, by a conventional method. Furthermore, dexylosylbenanomycin B obtained in free form may be converted into the corresponding salt thereof using the above-mentioned acid or base in a conventional manner.

Accordingly, the above-mentioned salts thereof as well as depsilocylbenanomycin B are included within the scope of the present invention. Further, when dexylosylpenanomycin B is reacted with lower alkanols such as alcohols such as methanol and elunol, the corresponding ester at its calcium group is produced.

Reference examples and working examples of the present invention are shown below, but since the properties of texiclosil penanomycin B have been clarified by the present invention, the method for producing tekilosil penanomycin B was based on those properties. Various methods can be devised. Therefore, the present invention is not limited to the examples, and the properties of dexylosyl penanomycin B, which are clearly improved by the present invention, can be modified by known means to obtain dexyloyl penanomycin B. Silbenanomycin B wo ff,
Covers all extraction and purification methods.

Reference Example 1 Actinobacteria MHI93-16 F4 cultured on agar slant medium
Strain (Feikoken Bacillus No. 9529) was mixed with starch 1. Inoculated into a liquid medium containing o-1 soybean flour 3.0 ml (500 - 80-1 pre-sterilized pi-17.0 in Yosaka Lof flask) and 2g0c
The culture was cultured with shaking (135 rpm) for 3 days to obtain a type 1 culture. Each of these 31 Rt cultured plates was inoculated into the same medium as above and cultured with shaking under the same conditions for 3 days to obtain a second type culture. 2L of this seed culture was inoculated into a 100A culture tank containing 50L of the medium with the above composition that had been sterilized at 120°C for 15 minutes,
2 days at 28″C, air flow 50L 7 minutes, 200 rpm
A third type culture was obtained by aeration and agitation culture at x. 125' in advance
Glycerin 2.0 '1, Nissanmi) 1.51, K2HP0.0.0025, KH
2P0.0.1125 Sorry, CoCl 2.6) 120
0.0005 yen, KM 720.03 yen, 57 containing 300 L production medium consisting of Adekanol 0.01 1
01 volume culture tank was inoculated with 12t of the above three types of fang culture, and 2g''
'C for 7 days with aeration and stirring at an aeration rate of 150 t/min for the initial 24 hours and 300 L/min and 1300 rpm for the subsequent 24 hours. After the culture was completed, diatomaceous earth was added as a filtration medium and the mixture was filtered to obtain 250 tons of filtrate (pH 6.0).

Reference Example 2 The culture filtrate 2501 obtained in Reference Example 1 was treated with Diaion H.
After adsorption on P-2015t and washing with water + 00t and 50% methanol 451, the active substance Y70j was eluted with methanol 45L1 and then methanol qot to obtain the OI fraction (
53L), second fraction (38t) and third fraction (274
) was obtained. The Kaede fraction containing the active substance was concentrated under reduced pressure.
When the value is adjusted to 3.5 using dilute hydrochloric acid, a red precipitate is obtained. This precipitate was collected by filtration and dried under reduced pressure, resulting in 152 g of brown coarse powder containing mainly benanomycin.
was gotten. 150 g of this crude powder was dissolved in dimethylformamide at 600 dK and saturated with water vapor in a desiccator at room temperature for 3 days to precipitate a crystalline precipitate. This precipitate was collected by filtration and dried under reduced pressure to obtain 29 g of penanomycin A (dimethylformamide sorbet). The second fraction was also treated in the same manner as the Otori fraction to obtain penanomycin A (dimethylformamide sorbet) 14J9.

Dimethyl sulfoxide (5-
) and added dropwise to 300 methanol with stirring,
After further stirring for 10 minutes, a reddish brown precipitate was deposited. This precipitate was collected by filtration and dried under reduced pressure to obtain pure benanomycin A reddish brown powder 935 Cape.

Reference Example 3 The third fraction obtained in Reference Example 2 was concentrated under reduced pressure to 1.5L.
When - was adjusted to 3.5 using dilute hydrochloric acid, a red precipitate was obtained. This precipitate was collected by filtration and dried under reduced pressure to obtain 99 g of brown coarse powder containing benanomycin B. This coarse powder 11 was heated in dimethylformamide 10 (4
0'C)fi[L, L filled with dimethylformamide
It was applied to a column of IL (-20) and developed with dimethylformamide.

Fraction 46 containing the active substance and 72 CI fraction 6-) were collected and concentrated to dryness under reduced pressure to obtain 657 η of brown crude powder containing benanomycin B dimethylformamide turbate. 300q of this crude powder was dissolved in methanol + 00ml, IN salt a1- was added thereto, and then concentrated to dryness under reduced pressure.

The obtained brown coarse powder was dissolved in dimethyl sulfoxide 3-, and added dropwise to 200-200 ml of chloroform while stirring.
After stirring for 0 minutes, a reddish brown precipitate was deposited. This precipitate was filtered and dried under reduced pressure to obtain pure penanomycin B hydrochloride 25811q as a reddish brown powder. In addition, the tube was sealed, and after reacting at 110°C for 12 hours, the precipitate deposited was filtered out. The obtained precipitate was extracted three times with dioxane IQ-,
Penanomycin (adalicon of benanomycin B) 47
．． I got 54. The residue was analyzed using preparative TLC (Merck Art
, 5744, developing system; butanol:acetic acid:pyridine:
Water = 6:l:4:3) to obtain fraction 1 containing dexlocurvenanomycin 7B and fraction 2 containing benanomycinone. Fraction - further/Iyaion HP-2
After adsorption at 0.20 mK and washing with water (60 m), elution was carried out four times with 40 methanol. The eluate was concentrated to dryness, and the residue was dissolved in 3 dK of water, and then adjusted to -2 with O,IN hydrochloric acid. When this solution is concentrated to dryness, pure dexylosylpenanomycin l hydrochloride+? , gW were obtained. Similarly, when fraction 2 was purified using Diaio7 HP-20, 9.5q of benanomycinone was obtained.

Example 2 lN MCI of Penanomycin B Hydrochloride 21719
-Methanol 40- The solution was sealed in a sealed tube and reacted at 90°C for 12 hours. Thereafter, the reaction solution was concentrated to dryness, the residue was adsorbed on Diamond Ion HP-20100tK, and after washing with water (300
mg) and extracted four times with 200 d of methanol. The extract was concentrated to dryness to obtain dexylosylpenanomycin B methyl ester 12519. The obtained ester is water 2
K I N NaOH 5- was added thereto and reacted at room temperature for 10 minutes. Then add IN hydrochloric acid 7 to the reaction solution.
- was added and concentrated to dryness. The residue is dissolved in water at 20 dK.
It was adsorbed on a column filled with 100- of Diaion MP-20, and after washing with water (300), it was washed with 200- of methanol.
Extracted twice. After concentrating the extract to dryness, Sephadex LH-
It was packed into a column packed with 20 parts of 6504 and developed with methanol. The active fraction was concentrated to dryness, and the remaining liquid was dissolved in 109I of water.
After dissolving in #t, it was adjusted to pt12 with IN hydrochloric acid. The solution was concentrated to dryness to yield 109.5 caps of pure dexylosylbenanomycin B hydrochloride as a reddish-brown powder.

(Effects of the Invention) As explained in detail above, dexylosylbenanomycin B or its salts and esters are obtained by chemical conversion of penanomycin B, and have antifungal activity to inhibit the growth of various molds.

[Brief explanation of the drawing]

f1 diagram: Dexylosylpenanomycin B hydrochloride in methanol (20 μf/ad) (shown by C solid line), 0, I
N-hydrochloric acid in methanol (20μt/g>C shown by broken line)
or o, +N*ell potassium in methanol (20
µf/seg) (indicated by the dashed line). Figure 2: Shows the infrared absorption spectrum of dexylosylbenanomycin B hydrochloride in potassium bromide tablets. Figure 3: Shows the 400 MHz hydrogen nuclear magnetic resonance spectrum of dexylosylbenanomycin B hydrochloride in deuterated dimethyl sulfoxide solution. FIG. 4: Shows the (F) IOQ MEz carbon nuclear magnetic resonance spectrum of dexlosylbenanomycin B hydrochloride in deuterated dimethyl sulfoxide solution. Procedures...Authentic (self-motivated) February 16, 1989

Claims (1)

[Claims] 1. Dexylosylbenanomycin B, an antifungal antibiotic, represented by the following formula ▲ Numerical formula, chemical formula, table, etc. ▼ (I), and its salt or ester. 2. A method for producing the anti-fungal antibiotic dexylosyl benanomycin B, which is characterized in that the anti-fungal antibiotic dexylosyl benanomycin B is produced by chemically converting the antibiotic benanomycin B.