JPS63162697A - Novel antibiotic substance sf-2415a3 and sf-2415b3 and production thereof - Google Patents

Abstract

NEW MATERIAL:Antibiotic substance SF-2415A3 having the following physical and chemical properties and its salt. Molecular weight, 520; molecular formula, C26H30N2O5Cl2; mass-spectrum (FD-MS), m/z=521(MH<+>); specific rotation, [alpha]D=+195 deg. (C=0.5, MeOH); solubility, soluble in methanol, chloroform, ethyl acetate, toluene and hexane and insoluble in water; color reaction, positive to molybdenum-sulfuric acid and 10% sulfuric acid reagent; appearance, red powder; etc. USE:An antibacterial agent. PREPARATION:A microbial strain belonging to Streptomyces genus and capable of producing SF-2415A3 and B3 substances [preferably Streptomyces sp.SF-2415 (FERM P-8801)] is cultured at 26-30 deg.C for 1-10 days preferably by submerged culture.

Description

DETAILED DESCRIPTION OF THE INVENTION Field of Industrial Application The present invention relates to novel antibiotic SF-2415A3 substance and S
This invention relates to F-2415B3 substances and their production methods.

Various antibiotics produced by microorganisms have been known in the past, but antibiotics similar to the antibiotics SF-2415A3 and SF-241583 according to the present invention include SF.
-2415A1, SF-2415A2. SF-2415
81 and SF-2415B2 [Patent Application 1986-19271
81. Napyranomycin
cin) 8, B1. B2. B3. C1 and C2 [
5hio et al.: J, ^ntibiotcs 39 (4
), 487'493 (1986) and J, ^ntib
iotics 39(4), 49j501 (1986)
]It has been known. However, the SF-241SA3 substance and the SF-241583 substance have different physical and chemical properties from these antibiotics, and are clearly distinguished from each other.

SUMMARY OF THE INVENTION The purpose of the present invention is to obtain a novel antibiotic useful for medical use.

As a result of our continued search for new useful antibiotics, the present inventors found a way to improve P.
The present invention was completed based on the discovery that novel antibiotic substances SF-2415A3 and SF-241583 having excellent antibacterial activity were produced in a culture of a certain strain belonging to the genus Streptomyces. That is, the present invention provides novel antibiotic substances SF-2415A3 and SF-2415B3, and methods for producing them.

Below are SF-2415A3 substances and SF-241583
The substances and their manufacturing methods will be specifically explained.

(1) Production bacteria and mycological properties of the production bacteria SF2415A3 substance and S used in the method of the present invention
The F-241583 substance production solution includes sufficient amounts of SF-2415A3 substance and SF to be collected into the culture.
-241533 substance may be used, but an example of such a strain is SF-2415 strain, which was newly isolated from the soil of Tottoriita by the present inventors. There is. The mycological properties of SF-2415 strain are as follows.

1. Morphology: The hyphae are long, well-branched, and do not divide under normal conditions. Aerial mycelium grows well on sucrose/nitrate agar, glycerol/asparagine agar, starch agar, etc., and spore formation is abundant.

The aerial hyphae are simply branched, and JJI pongee branching is not observed.

The spore chain at the tip of the aerial hyphae is mainly spiral-shaped.

When observed using an electron microscope, the spores are oval in shape and have a diameter of 0.8-1.
．． 2X1. It has a size of O-1,6μ-, and the surface is spiny or warty.
Spores usually form chains of 10 or more spores. No sporangia, sclerotia, or flagellum were observed.

(2) Growth status on various media The growth status of SF-2415 strain on various media is shown in the following table. Regarding the color description [The standard shown in 1 is Container/Corporate/Eve/America (Container Co., Ltd.)
tainer Corporation of＾mer
ica) Manufactured by Color Fist Harmony ◆Manual (C.

For llarmony Manual) J. Observations were made after culturing at 28°C for 14-21 days.

■Physiological properties (1) Growth temperature range: 15% on yeast/malt agar
Overcoat in the temperature range -37°C and lhy'r well at 26-30°C.

(2) Liquefaction of gelatin: Positive (3) Hydrolysis of starch: Positive (・t) Reduction of nitrate: Negative (5) Peptonization of skim milk: Negative Coagulation of skim milk: Negative (6) Salt tolerance: 3% It is grown in a medium containing 4% salt.
It does not grow north of this area.

(7) Melanin-like pigment production I&: negative ■, carbon source availability: (Pridba+i and (:oL
Llicb basal medium [heavy use] (1) Use: D-glucose, D-7 lactose, D-xylose, 1. -7 rabinose, D-mannitol, lafi7-se, L-ram7-su (2) Not used: i-i7 citol, shekrose■
, cell wall composition method of Becker et al. [^ppl, Mie
robio1.13e236, (1965)], the cell wall composition components No7mi7 and Melinic acid were of the LL type.

Based on the above properties 1), it is reasonable to believe that strain SF-2415 belongs to the genus Streptomyces among actinomycetes.

Therefore, the present inventors used the SF-2415 strain as Streptomyces sp.
yces sp, SF-2415).

Strain SF-2415 has been entrusted to the Institute of Microbial Technology, Agency of Industrial Science and Technology as Microtechnology Laboratory Moeyori No. 8801 (FEBN P-8801).

The properties of this bacterial strain, SF-2415 strain, tend to change as seen in the case of actinomycetes in ponds. for example,
The SF-2415A3 substance and the SF-2415A3 strain, even if it is a mutant strain (naturally occurring or induced), a phenozygote, or a genetically recombinant strain derived from the SF-2415 strain.
All Streptomyces bacteria capable of producing the 241583 substance can be used in the method of the present invention.

(2) Cultivation method In the method of the present invention, the above-mentioned bacteria are cultured in a medium containing nutrients that can be used by ordinary microorganisms. As a source of nutrients,
It can be made using glucose, starch syrup, dextrin, sucrose, starch, molasses, animal/vegetable oils, etc. Also, as a nitrogen source, soybean flour, wheat germ, cornstarch liquor, cottonseed waste, meat extract, peptone, yeast extract, ammonium sulfate, sodium nitrate, urea, etc. can be used. The pond, as necessary, sodium, potassium, calcium,
It is effective to add Kuroiso salts, which can generate magnesium, cobalt, chlorine, phosphoric acid, sulfuric acid, and other ions. Also helps seedling growth, SF2415
Materials and non-organic substances that promote the production of A3 substance and SF-241583 substance can be appropriately added.

As a culture method, a culture method under aerobic conditions, especially a deep culture method is most suitable. The appropriate temperature for culturing is 15-37℃.
But in many cases. Culture at around 26-30°C.

The production of SF-2415A3 substance and Sl:'-241583 substance varies depending on the medium and t5=1 conditions, but the accumulation usually reaches its maximum within 1 to 10 days in both shaking culture and tank culture. When the accumulation amount of SF-2415A3 substance and SF-241583 substance in the culture reaches the maximum, the culture is stopped, and the target substance is isolated and purified from the culture solution.

(3) Purification SF-2415A3 substance and S obtained by the present invention
To collect the F-241583 substance from a culture, use conventional separation methods that utilize its properties, such as solvent extraction, ion exchange resin, adsorption or distribution column chromatography, gel filtration, dialysis, and precipitation. These can be extracted and purified singly or in appropriate combination. For example, SF-
The 2415A3 substance and the SF-241583 substance are extracted from the cultured bacterial cells with acetone-water or methanol-water. Further, the SF-2415A3 substance and the SF-2415B3 substance accumulated in the culture solution are adsorbed to a synthetic adsorbent such as Diaion HP-20. In addition, if extracted with an organic solvent that does not mix with water, such as ethyl acetate or chloroform, SF-2415A3 substance and SF-241
The 583 substance is extracted into the growth solvent layer.

To further purify the SF-2415A3 substance and the SF-241583 substance, silica gel (Wako Gel C-30
0, manufactured by Wako Pure Chemical Industries, Ltd.), alumina, or Sephadex LH-20 (7 Furumashi 7 Company 91).

SF-2,11s produced in this way in culture
A3 Substance SF-241583 Substance can be isolated as a free form, and SF-2415A3 Substance and SF
-2415B3 substance-containing solution or its concentrate with a base, that is, an alkali metal compound such as sodium hydroxide, potassium hydroxide, etc., an alkaline earth metal compound such as calcium hydroxide, magnesium hydroxide, etc.
When treated with a Kuroiso base such as ammonia, for example, an organic base such as ethanolamine, triethylamine, dicyclohexylamine, etc. during the fabrication of each step, for example during each step of extraction, separation or purification, SF-24
The 15A3 substance and the SF-2415B 3 substance are converted into their corresponding salt forms and separated. In addition, the SF-2415A3 substance and SF produced in this way
-241583 substance can be prepared in free form by conventional methods, i.e., SF-2415A3 substance and SF-241583 substance.
It can be easily transformed into the substance itself.

Furthermore, the SF-2415A3 substance and the SF-241583 substance obtained in free form may be converted into the corresponding salts thereof using the base in a conventional manner.

Therefore, SF-2415A3 substance and SF-241583
The substances as well as their salts as described above are intended to be included within the scope of this invention.

The SF-2415A3 substance and SF-241583 substance obtained according to the above production method have the following physical properties.

1. Physicochemical properties of antibiotic SF-2415A3 substance ■Molecular weight and molecular formula % Formula % ■Infrared absorption spectrum (KBr method, am-') 342
0, 2980.2920.2850.2160.214
0°1690, 1650.1615.1515.143
0.1390°1370, 1270.1165.112
0.1090.800, 750 [5] Ultraviolet absorption spectrum Eλwax n+s (ε) 1254 (18300)
, 302 (18900).

376 (4900), 450 (3800) 252 (1
3400), 301 (19200).

375 (5200), 442 (3800)■'HNM
R spectrum (400MH2) in deuterated chloroform solution,
Measurements were made using TMS as a reference substance.

δ (ppm)', 11J6 (IL s) + 4.93
(IL mL4.77 (III, br dd), 4
．． 40(ill, dd).

2.75 (LH, br dd), 2.72 (11 (,
brdd).

2.64 (IH, dcl), 2.53 (III, d
d).

2.12 (311, s) = 1.74 (411, br
s).

1.64 (311, br s), 1.51 (6H, b
r 5) tl, 38 (311, br d), 1.19
(3H,s)■11eNMR vector (100MHz
) Measurement was carried out in a deuterated chloroform solution using TMS as a reference substance.

δ (ppm): 192.9 (s), 192.2 (s)
, 173.2(s).

159.8(s), 143.2(s), 133.9
(s).

131.7(s) = 123.1(d), 122.9
(8).

114.4(d), 112.8(s), 83.5(s
).

80.8(S), 79.1(S), 78.3(3)
, 58.2(d) 42.9(t), 41°7(t)
, 39jl(t), 28.8(q)26.3(t)
, 25.8(q), 22.5(q), 17.7(
q) 16.7 (q) = 9.3 (q) ■ Solubility Soluble in methanol, chloroform, ethyl acetate, toluene, hexane. Insoluble in water.

■ Color and properties of the substance Red powder [Phase] Color reaction Molybdenum-sulfuric acid, positive for 10% sulfuric acid reagent ■ Thin layer chromatography carrier Nijiri Cadel plate (manufactured by Merck & Co., Ltd.) Development) 8 medium system R1 value Toluene-ethyl acetate ( 2:1) 0067 Hexane-acetone (3:1) 0.53 Chloroform-Methyl (50:1) l') , 70 ■, Physicochemical properties of antibiotic SF-2415B3 substance ■ Molecular weight and molecular formula % Formula % ■Infrared absorption spectrum (KBr method, cm') 3400
, 2980.2920.2850.1685.1640
゜1600, 1500.1430.1390.1370
．． 1350°1305, 1,280.1120.108
0.930.800.770, 750 [5] Ultraviolet absorption human vector (λ1x stop (ε))” eO■: 205 (
21400>, 266 (20900>.

8 m a x 327 (7800), 360 (7400) AHe””L
: 205 (16600), 267 (21600).

+11aX 332 (7900), 356 (7600) ■' HN
MRS (400 MHz) Measurement was carried out in a deuterated chloroform solution using TMS as a reference substance.

δ(111111): 12,14(1111s),
7.33 (IL s) + 6.78 (1111br s)
+ 4.87 (111+ mL4.70 (IH, br
L), 4.43 (III, dd) 12.7H2H, b
r d), 2.48 (IH, dd).

2.43 (ltl, dd), 2.22. (3H,
5) -1,62(311,br s), 1.60(2
11, Zeng).

1.58 (2H, m), 1.51 (3fl, s).

1.48(3H,br d), 1.38(3H,br
d).

1, 18 (311, s) 13C NMR spectrum (100 MHz) Measured in a deuterated chloroform solution using TMS as a reference substance.

δ (ppm): 196.8 (sL 193.4 (s)
, 162.1(s).

161.3(s), 142.3(s), 131.5
(s).

131.4(s), 123.4(d), 120.3
(s).

114.6(d), 109.4(s), 107.0
(d).

83.4(s), 79.1(s), 78.7(s),
58.8(d)42.9D), 41°4(t),
39.8 (t), 28.9 (q) 26.1 m,
25.8(q), 22.4(q), 17.7(q
) 16.7(q), 8.6(q) ■ Solubility Soluble in methanol, chloroform, ethyl acetate, toluene, hexane. Insoluble in water.

■ Color and properties of the substance Yellow needle-like crystals [phase] Color reaction Molybdenum-sulfuric acid, positive for 10% sulfuric acid reagent ■ Thin layer chromatography carrier Nijiri Cadel plate (manufactured by Merck & Co.) IJ1 open solvent system Rf value Toluene-ethyl acetate (2:1) 0.65 hexane-77 tons (3:1) 0.45 chloroform-methanol (50:1) 0. SF-2415A3 (1) and SF-241 according to the physical properties of the first half of 51
The structure of the 583(n) substance was deduced as follows.

(1) (n) Examples of the present invention are shown below, but these are merely examples and do not limit the present invention.

Of course, many variations or modifications not exemplified here may be used.

Example 1゜Seed medium, starch 2.0%, glucose 1゜0%
, wheat embryo 0.6%, polypeptone 0.5%, yeast extract 0.3%, soybean flour 0.2%, calcium carbonate 0.
A medium containing 1% was used. In addition, as a production medium, starch 2.0%, soybean oil 1.0%, cottonseed meal 1.5%, corn gluten meal 0.7%, calcium carbonate 0.3%,
A medium containing 0.001% of ferrous sulfate (NL heptahydrate) was used. In addition, all pifs before sterilization were adjusted to P117.0 before use.

A 100 e+l triangular 7 rasp containing 20 volumes of the seed medium was sterilized at 120°C for 30 minutes, and Streptomyces sp. SF-2415 (FERN P
-8801) was inoculated with 1-2 platinum loops of slant culture, and 28
The cells were cultured with shaking at ℃ for 3 days to form the first type culture. Then, the 50-1 volume Erlenmeyer flask into which 80-mL of the seed medium was dispensed was heated to 120°C.
Sterilize for 30 minutes at
The cells were cultured with shaking for 2 days, and this was used as a second type culture. Furthermore, 7 5L triangular 5L volumes of seed medium IL were dispensed at 120°C.
Sterilize for 30 minutes, inoculate with 150 ml of No. 21 culture, and
The cells were cultured with shaking at 8° C. for 1 day, and this was designated as the 3rd iI+ culture. A 50L jar containing 35L of production medium previously sterilized for 30 minutes at 120°C and a 7-armenter was inoculated with 1IL of the above T53 seed culture, aerated at 28°C for 3 days (2017 minutes),
Stirring (initial 250rpm+, 400rpm after 41 hours)
pm) was cultured.

After culturing, diatomaceous earth is added as a filter aid and filtered.
It is not possible to obtain 30 L of filtrate and 10 L of compressed cells.

Example 2゜30L of the culture medium 11 filtrate obtained in Example 1 was adsorbed on 3L of Diaion HP-20 (Mitsubishi Kasei Corporation).
The active ingredient was dissolved in 0.5N ammonium hydroxide-acetone (1
:1) (30L). The eluate containing the active ingredient is concentrated under reduced pressure to a volume of 5 L. At the same time, 77 tons of water (1:
1) Extract the active ingredient using 30 L, filter the extract from the bacterial cells, and concentrate under reduced pressure to a volume of 10 L. 5 L of the concentrate obtained previously and 10 L of the bacterial cell extraction concentrate were combined to make 15 L, and the active ingredient was extracted with 15 L of ethyl acetate. The extracted ethyl acetate solution was concentrated under reduced pressure to obtain 38 g of an oily substance.

Example 3゜The oily substance 38Fi obtained in Example 2 was mixed with silica gel 50
After thoroughly drying under reduced pressure, apply a silica sol column 400
Filled to the top of ml. Developing solvent: toluene-ethyl acetate/l, (7s: 1)300a+1(fr, 1-2
), toluene-ethyl acetate (50:1) 340 1
(ft, 3-4), toluene-ethyl acetate (20:
1) 1000+al(fr, 5-10) and toluene-ethyl acetate (10:1) 11001Ill(
When expanded in the order of fr, 1l-16), the active area mainly containing the SF-2415B2 substance is r r,
SF-241581 and SF-2,115B3 on 6-7
The active zone mainly containing the substance is rr.

The active area mainly containing SF-2415B1 and SF-2415A2 substances is fr in 8, and SF''-241 in 9-11.
The active area is mainly 5A2 and SF''-2415A3 substances,
An active zone mainly containing SF-2415A1 substance was separated and obtained at fr, 13-15.

Example 4 Two fractions containing mainly the SF-2415B2 substance obtained in Example 3 were concentrated under reduced pressure to obtain 1.26 g of crude material.

The crude material was sprinkled onto 10 g of silica sol, placed on top of a 300 ml silica gel column prefilled with toluene, and eluted with a developing solvent of toluene-ethyl acetate (20:1). Of the elution fractions, both fractions containing the SF-241582 substance were collected and concentrated to dryness to yield 180 μ of SF-2415B.
Two substances were obtained.

Example 5゜SF-2415B1 and SF-24 obtained in Example 3
When both parts containing mainly 1583 substances are concentrated under reduced pressure, the result is 1.09
g of crude material was obtained. This crude material was dissolved in a small amount of a mixture of meta/-chloroform (1:9), and this was poured into a Sephadex L H-20 (S
The product was subjected to column chromatography using OOml), developed with a methanol-chloroform (1:9) mixture, and the active fractions were collected and concentrated under reduced pressure. Furthermore, the obtained crude substance is 0
．． The mixture was sprinkled with 5 g of silica sol and placed on a 50-millimeter silica gel column that had been filled with toluene in advance. Toluene-
The active substance was eluted with a developing solvent of ethyl acetate (75:1), and the two fractions containing the SF-2415B1 substance were collected and concentrated to dryness, yielding 280 g of the SF-241581 substance and the SF-2415B3 substance. When the two components contained were concentrated and dried, 25011 g of SF-2415B3 substance was obtained.

Example 6゜SF-2415A2 and SF-24 obtained in Example 3
When both parts containing mainly 15A3 substance are concentrated under reduced pressure, the result is 1.16
g of crude material was obtained. This crude material was dissolved in a small amount of a mixture of methanol and chloroform (1:9), and this was poured into a Sephadex L H-20 (
The product was subjected to column chromatography using SOOml) and developed with a mixture of meta-7-chloroform (1:9), and the active fractions were collected and concentrated under reduced pressure. Further, the obtained filtrate was added to 0. It was sprinkled on silica gel of SF1 and placed on top of a silica gel column 40×1 filled with toluene in advance. The active substance was eluted with a developing solvent of toluene-ethyl acetate (75:1) and then toluene-ethyl acetate (so:i), and both fractions containing the SF-241SA2 substance were collected and concentrated to dryness, yielding 54H SF-2. , 115A2 substance was obtained. Both fractions mainly containing the SF-2415A3 substance were concentrated under reduced pressure, and then subjected to preparative TLC (Merck ^rt, 5
744. Purification with R amount hexane-acetone (3:1) yielded 50 g of SF-2415A3 material.

Example 7 Two fractions containing mainly the SF-2415A1 substance obtained in Example 3 were concentrated under reduced pressure to obtain 1.14 g of crude material.

This crude material was mixed with a small amount of methanol-chloroform (1:9
), and this was subjected to column chromatography using Sephadex LH-20 (80 (LEL)) filled with the same mixture in advance, and developed with a mixture of meta/-chloroform (1:9). When the active fractions were collected and concentrated under reduced pressure, $1 containing mainly 31hg of SF-2415A1 substance was obtained.
Substance obtained. This crude material was sprinkled on 0.5 g of silica sol, and a silica gel column 60 was filled with toluene in advance.
Place it on top of the ml. Toluene-ethyl acetate (10:
Elute the active substance with the developing solvent of 1) and add it to SF-2415A.
When fractions containing one substance are collected and concentrated to dryness, 132H
SF-2415A1 substance was obtained.

Effects of the Invention The SF-2415 substance according to the present invention exhibits activity against Durham-positive bacteria, and is expected to be useful as an antibacterial agent. Table 1 shows the minimum inhibitory concentration for each test bacterium.

S. enLeritidis No. 11 > 1
00 >100

[Brief explanation of the drawing]

Figure 1: Shows the infrared absorption spectrum of SF-2415A3 substance in potassium bromide tablets. Figure 2: Shows the infrared absorption spectrum of SF-241583 substance in potassium bromide tablets. Figure 3: Shows the ultraviolet absorption spectrum of SF-2415^3 substance in methanol. Figure 4: Shows the ultraviolet absorption spectrum of SF-2415^3 substance in hydrochloric acid-methanol. Figure 5: Ultraviolet absorption spectrum of SF-241583 substance in methanol. Figure 6: SF-241583 substance hydrochloric acid-methanol:
The ultraviolet absorption spectrum in 8 liquids is shown. Figure 7: Shows the 400 MIIz hydrogen nuclear magnetic resonance spectrum of SF-2415^3 substance in deuterated chloroform. Figure 8: Shows the 400 MIIz hydrogen nuclear magnetic resonance spectrum of SF-241583 substance in deuterated chloroform. Figure 9: Shows the carbon nuclear magnetic resonance spectrum of SF-2415A3 substance at 100 MIIz in deuterated chloroform. Figure 10: Shows the carbon nuclear magnetic resonance spectrum of SF-241583 substance in deuterated chloroform at 100 MIIz.

Claims (4)

[Claims] (1) SF-, a novel antibiotic with the following physical and chemical properties
2415A3 substance and its salt. [1] Molecular weight and molecular formula 520, C_2_6H_3_0N_2O_5Cl_2 [2] Mass vector (FD-MS) m/z 521 (MH^+) [3] Specific optical rotation [α]_D+195° (c0.5, MeOH) [4] Infrared absorption Spectrum (KBr method, cm^-^1
) 3420, 2980, 2920, 2850, 2160,
2140, 1690, 1650, 1615, 1515,
1430, 1390, 1370, 1270, 1165,
1120, 1090, 800, 750 [5] Ultraviolet absorption spectrum [λmax nm (ε)] λ^M^e^O^H_m_a_x: 204 (19500
), 240 (16900, sh), 254 (18300
), 302 (18900), 376 (4900), 45
0 (3800) λ^M^e^O^H^-^H^C^I:_m_a_x2
04 (14900), 240 (17500, sh) 25
2 (18400), 301 (19200), 375 (5
200), 442 (3800) [6]^1H NMR spectrum (400 MHz) Measured in a deuterated chloroform solution using TMS as a reference substance. δ (ppm): 11.36 (1H, s), 4.93 (1
H, m), 4.77 (1H, br dd), 4.40 (
1H, dd), 2.75 (1H, br dd), 2.7
2 (1H, br dd) 2.64 (1H, dd), 2.
53 (1H, dd), 2.12 (3H, s), 1.74
(4H, br s), 1.64 (3H, br s), 1
．． 51 (6H, br s), 1.38 (3H, br d
), 1.19 (3H, s) [7]^1^3C NMR spectrum (100MHZ) Measured in deuterated chloroform solution using TMS as a reference substance. δ (ppm): 192.9 (s), 192.2 (s),
173.2 (s), 159.8 (s), 143.2 (s
), 133.9 (s), 131.7 (s), 123.1
(d), 122.9(s), 114.4(d), 112
．． 8(s), 83.5(s), 80.8(s), 79.
1(s), 78.3(s), 58.2(d), 42.9
(t), 41.7(t), 39.8(t), 28.8(
q), 26.3(t), 25.8(q), 22.5(q
), 17.7(q), 16.7(q), 9.3(q) [8] Soluble Soluble in methanol, chloroform, ethyl acetate, toluene, hexane. Insoluble in water. [9] Color and properties of the substance Red powder [10] Color reaction Molybdenum-sulfuric acid, positive for 10% sulfuric acid reagent [11] Thin layer chromatography carrier: Silica gel plate (manufactured by Merck & Co.) Developing solvent system Rf value Toluene-acetic acid Ethyl (2:1) 0.67 Hexane-acetone (3:1) 0.53 Chloroform-methanol (50:1) 0.70 (2) Novel antibiotic SF- with the following physical and chemical properties
2415B3 substance and its salt. [1] Molecular weight and molecular formula 494, C_2_6H_3_2O_5Cl_2 [2] Mass vector (FD-MS) m/z 494 (M^+) [3] Specific luminosity [α]_D+33° (c 0.5, MeOH) [4] Infrared rays Absorption spectrum (KBr method, cm^-^1
) 3400, 2980, 2920, 2850, 1685,
1640, 1600, 1500, 1430, 1390,
1370, 1350, 1305, 1280, 1120,
1080, 930, 800, 770, 710 [5] Ultraviolet absorption spectrum [λ max nm (ε)
] λ^M^e^O^H_m_a_x: 205 (21400
), 266 (20900), 327 (7800), 36
0 (7400) λ^M^e^O^H^-^H^C^l_m_a_x:2
05 (16600), 267 (21600), 332 (
7900), 356 (7600) [6]^1H NMR spectrum (400MHZ) Measured in deuterated chloroform solution using TMS as a reference substance. δ (ppm): 12.14 (1H, s), 7.33 (1
H, a), 6.78 (1H, br s), 4.87 (1
H, m), 4.70 (1H, br t), 4.43 (1
H, dd), 2.71 (2H, br d), 2.48 (
1H, dd), 2.43 (1H, dd), 2.22 (3
H, s), 1.62 (3H, br s), 1.60 (2
H, m), 1.58 (2H, m), 1.51 (3H, s
), 1.48 (3H, br d), 1.38 (3H, b
r d), 1.18 (3H, s) [7]^1^3C NMR spectrum (100 MHz) Measured in a deuterated chloroform solution using TMS as a reference substance. δ (ppm): 196.8 (s), 193.4 (s, 1
62.1(s), 161.3(s), 142.3(s)
, 131.5(s), 131.4(s), 123.4(
d), 120.3(s), 114.6(d), 109.
4(s), 107.0(d), 83.4(s), 79.
1(s), 78.7(s), 58.8(d), 42.9
(t), 41.4(t), 39.8(t), 28.9(
q), 26.1(t), 25.8(q), 22.4(q
), 17.7 (q), 16.7 (q) 8.6 (q) [8] Soluble Soluble in methanol, chloroform, ethyl acetate, toluene, hexane. Insoluble in water. [9] Color and properties of the substance Yellow needle crystals [10] Color reaction Molybdenum-sulfuric acid, positive for 10% sulfuric acid reagent [11] Thin layer chromatography carrier: Silica gel plate (manufactured by Merck & Co.) Developing solvent system Rf value toluene -Ethyl acetate (2:1) 0.65 Hexane-acetone (3:1) 0.45 Chloroform-methanol (50:1) 0.51 (3) Streptomyces
) SF-2415A3 substances belonging to the genus SF-241
SF-2415A3 substance and/or SF-2415B3, characterized in that 5B3 substance-producing soda is cultured in a medium and the compound or salt thereof according to claims 1 and 2 is collected from the culture. Method of manufacturing a substance. (4) Streptomyces
) The producing bacteria belonging to the genus Streptomyces sp.
The manufacturing method according to claim 3, which is F-2415 (Feikoken Kyoiku No. 8801).