JPS58116485A - Novel antibiotic carbapenem derivative and its preparation - Google Patents

Abstract

NEW MATERIAL:3-Hydroxymethylsulfinyl-6- ( 1-hydroxysulfonyloxyethyl ) -7-oxo- 1-azabicyclo[3,2,0]-hept-2-ene-2-carboxylic acid shown by the formula and its salt. USE:An antibiotic. Having a wide range of antibacterial spectrum and-lactamase inhibiting action. PROCESS:A bacterium capable of producing pluracidomycin C is subjected to liquid culture under aerobic conditions in a nutritive medium at 20-40 deg.C, preferably at 25-32 deg.C in pH of the medium of 5.5-8.5, and a compound shown by the formual or its salt is separated and collected from the culture after the cultivation is over. The compound or its salt is reduced with sodium boron hydride, etc. to give a compound having increased objects of effect.

Description

[Detailed description of the invention] Ru.

The antibiotic provided by the present invention has a broad antibacterial spectrum and β
- A new stable carbapenem antibiotic with lactamase inhibitory action.

3-Hydroxymethylsulfinyl-g-(l-hydroxysul-nyloxyethyl)-7-oxo-noazabicyclo[3.2, θ]-hebut-2-ene-λ-monocarboxylic acid. Its structural formula is shown below.

The present invention encompasses the two above-mentioned antibiotics and their pharmaceutically acceptable salts. Examples of the salt include salts with alkali metals (sodium, potassium, etc.), alkaline earth metals (calcium, barium, etc.), and the like.

Carbapenem antibiotics are produced by actinomycetes.

Various methods are known, but in equation (1), c6(
1'b-8-CH=CHNHCOCH, MM/711
0 (L=-8-CH,-CH,-NHCOCH
The compound is known in the literature.

9 Prior to the present invention, the inventors have developed the above-mentioned mother nucleus (CHO)
was found to have antibacterial and β-lactamase inhibitory effects (Patent Application Shoj, 4; -/701>27)
A-g17g6-c).

The present inventors have now completed the present invention by discovering that by reducing pullacidomycin C, a compound having antibacterial activity and an increased number of targets for β-lactamase inhibitory activity can be obtained.

In the antibiotic of the present invention, in formula (1), R is -S-CH
, OH, and is a novel antibiotic that is completely different from known carbapenem antibiotics in which R is a cystinoamine derivative.

The physical properties of this antibiotic sodium salt are shown below.

(a) Infrared absorption spectrum Br νmax 3320.21 0, /770, /1
,20. /! ;90°lda00. /230. /ko3θ,
lθ65. /θ20°93j, 93.773. Otsu 2J
,! ; and jα−′(b)′H-nuclear magnetic resonance vector (
In heavy water, external reference TMS) (J=Hz) δ 197C3H, d, J, , -Otsuj), 3.37C
/H,d-d.

J=/OJ and l and 3), 3.90(/H, d=
d, J=93 and /i,! ;), lA3-4/J4B
3H, m).

Hiro 9! ; (/H, m), JJg (/H, m) ppm Next, 9 methods for producing the antibiotic of the present invention will be shown. This is carried out by culturing burracidomycin C-producing bacteria in a nutrient medium under aerobic conditions, and after completion of the cultivation, burracidomycin C and its salts are separated and collected from the culture, and then reduced.

As for the culture medium composition, culture conditions, etc., those generally used in the production of antibiotics may be used. In principle, the medium is
Contains carbon sources, nitrogen sources, inorganic salts, etc. If necessary, vitamins, precursors, and others may be added to increase the production of plracidomycin C. Examples of carbon sources include glucose.

Starch, texturin, glycerin, molasses, organic acids, etc. may be used alone or in mixtures. Examples of nitrogen sources include soybean flour, corn steep liquor, meat extract,
Yeast extract, cottonseed flour, peptone, wheat germ, ammonium sulfate, ammonium nitrate, etc. are used alone or in mixtures.

Examples of inorganic salts include calcium carbonate, sodium chloride, potassium chloride, and magnesium sulfate.

Examples include cobalt chloride and various phosphates, which are added to the culture medium as necessary.

Cultivation may be carried out according to methods generally used for the production of antibiotics, and liquid culture, particularly deep aeration culture, is preferred when mass production is performed. The pH of the medium is preferably about 1j-13, and the culture temperature is about 20 to θ°C, especially 25 to 32°C.
is preferred. Although the culture time largely depends on the scale of fermentation, the culture time for mass production is approximately 20 to 0 hours.

After completion of the culture, to separate and collect Plracidomycin C and its salt from the culture, a conventional method for separating and collecting fermentation products is appropriately used. for example.

Filtration, centrifugation, adsorption and desorption using various ion exchange resins and other active adsorbents, and chromatography.

It is advisable to appropriately combine extraction with various organic solvents. Furthermore, in order to prevent decomposition of Plracidomycin C and its salts, it may be considered to use a suitable stabilizer as necessary during one separation step.

The reduction reaction may be carried out in the presence of a reducing agent, and examples of the reducing agent used at this time include lithium borohydride, sodium borohydride, potassium borohydride, etc. In particular, sodium borohydride is used. preferable. The reaction is carried out at room temperature for 7 to 2 hours at a pH of 6 to 7, preferably 7.
Let's do it. As a solvent, water or methanol,
Alcohols such as ethanol, tetrahydrofuran,
It is preferable to use a water-soluble solvent such as dioxane.

To separate and purify the target antibiotic after the reaction is complete.

Commonly used methods such as filtration, centrifugation, and chromatography may be used.

Strepto-myc ticus, a plurasidomycin C-producing bacterium.
es p, Iuracidomyceticus) is
It is an actinomycete that was isolated from a soil sample in 1970, and the standard strain was sent to the Microbial Technology Research Institute in Yatabe-cho, Tsukuba-gun, Ibaraki Prefecture from February 1st, 1981, under the name J9 Otsu. It has been deposited as. Its mycological properties are as follows:
It is described in θ621r.

In the present invention, the above-mentioned strains can of course be used.

All bacteria belonging to the genus Streptomyces and producing the antibiotic plurasidomycin C can be used.

The antibiotic of the present invention has a broad antibacterial spectrum, is effective against both Durum-positive bacteria and Durum-negative bacteria, and is effective against Enterobacter cloacanita 2 (Enterobacter cloacanita 2).
ter cloacae 9,2)
0 sporinase-type and penicillinase-type β-lactamases and Klebsiella sp. 363 (Klebsiella sp.
It has a strong inhibitory activity against penicillinase-type β-lactamase produced by S. siella Sp, 31J), and is therefore useful as a medicine, veterinary medicine, and disinfectant. The antibacterial spectrum of this antibiotic is shown below.

(See margin below) Therefore, the antibiotic of the present invention can be administered to humans or animals orally or parenterally. 9 tablets using commonly used excipients, stabilizers, preservatives, wetting agents, surfactants, etc.
It can be administered orally in capsules, powders, etc., as well as injections and liniments.

It can also be administered parenterally, such as in half.

Although the dosage varies depending on the therapeutic purpose, it is generally about 1/10 to several times the amount of cephalothin.9 For example, in the case of subcutaneous administration, the daily dose for an adult is about 0.1 to 30 g.

Note that since the two antibiotics have a β-lactamase inhibiting effect, the antibacterial activity of the β-lactam antibiotic against β-lactamase-producing bacteria can be increased synergistically. Accordingly, known β-lactam antibiotics 1, for example benzylpenicillin.

Phenoxymethylpenicillin, carbenicillin.

Penicillin antibiotics such as ampicillin and amoxicillin, cephaloridine, cephalothin, cefatholin, cephalexin, cefoxitin, cefacedryl, cefamandole, and cephapirin.

Cefrazine, cephaloglycin, ceftesol.

It may be used in combination with cephalosporin antibiotics such as cefatridine and cefmetazole.

Next, an example of producing the target compound of the present invention will be shown.

The following examples are not intended to limit the invention in any way.

Reference example (a) Fermentation process: soluble starch 05%, glucose 03%
, polypeptone θj% + beef extract θj% yeast extract θ
Streptomyces pluracidomyceteicus (
A seed culture slant of FERM-PJ-, 9 Og) was inoculated and cultured with shaking at Iθ rotations per minute for 2t'c and gr hours. Add 100ttl of this culture solution to 2.11% tomato paste, 2.4g% texturin, dry yeast/
Part 2, Cobalt chloride hydrate θθ0θOtchi, water (pH 7
0, before sterilization), was inoculated into a 3θl capacity jar fermenter containing 20A, and the aeration rate was λθl/min, and the internal pressure was 0.
2 kg/cyr'G.

Stirring number/30~3! ; Incubate in OrPm for 2f'C and 2S hours.

(b) Separation step: The culture solution obtained in the above step
), benzyldimethyl-cetyl ammonium chloride (
Methylene chloride (4Zθ) containing 72%) was added to transfer the active substance to the methylene chloride layer.

This methylene chloride layer was dissolved in an aqueous solution of sodium trithiiodide (3
By back-extracting in step 1 and freeze-drying, a crude sample of the active substance (O θg) is obtained. This crude specimen powder (〃g) was gel-filtered through 600 ml of Biogel P-2 (manufactured by Bio-Rad) using water as the mobile phase, and the fractions showing activity against E. coli were collected and freeze-dried. Product C1779) is obtained. This active fraction preparation (3 g) was added to QAE-Sephatex A.
-23 (manufactured by Pharmasha), when subjected to Gradendo C003%-03 of common salt with oOj ammonium chloride and adjusted to pH 7θ, C fraction (29θg/
), B fraction Cl30m/), A fraction (330ttr
l) will be eluted in this order. Fraction containing C (approximately 3θm
l) was desalted using a Biokel P-2 column.

The fractions containing the active substance are collected, concentrated at a pH of 2%, concentrated under reduced pressure, and lyophilized to obtain a powder of plracidomycin C sodium salt (/j■).

Example 003; M') The powder of Pluracidomycin C sodium salt obtained in the separation step C1011fl) was added to an acid buffer solution (pH 003;M').
70.3.1 ml) and stirred at room temperature. ; 611f, l/3 molar equivalent) in water (26 tμl) is added.

After 70 minutes, add and dissolve sodium chloride (ly),
Adjusted with an aqueous sodium chloride solution, the t-codia ion H
p-1-2oha <ioo ~ 2θ0 mesh) (manufactured by Mitsubishi Kasei Corporation) with a 2.0 resistant column) was eluted with an aqueous solution of IOS sodium chloride, and! Fractionate by 1 tnl.

Fraction f where activity is detected by high performance liquid chromatography
-Collect #; below 23;'C.

Concentrate to 3 ml under reduced pressure to remove precipitated sodium chloride. Sephadex G-10Ctθ~/2 for furnace liquid
Elute with water and desalt. Transfer the desalted fraction to a 300 t column of 0μ).

Concentration to approximately 3 ml under reduced pressure and lyophilization yields 3-hydroxymethylsulfinyl-(/-hydroxysulfonyloxyethyl)-7-oxo-l-azabicyclol120]- as a colorless amorphous powder <rv). Hebut-2-ene-2-carboxylic acid is obtained.

Patent Applicant: Shionogi & Co., Ltd. Procedural Amendment ζ Spontaneous) ] Otori ＼＼Showaman, September 7, 1949 Director General of the Patent Office / Case Indication 1982 Patent Application No. 2/24t3j;
No. 2 Name of the invention Novel antibiotic carbapenem derivatives and their manufacturing method 3 Relationship with the amended case Patent applicant address 3-12 Doshomachi, Higashi-ku, Osaka-shi, Osaka Prefecture Name (
/92) Shionogi & Co., Ltd. Representative Yoshitoshi
- Yuhiro Agent Address No. 12-q Sagisu J-chome, Fukushima-ku, Osaka Shionogi & Co., Ltd. Patent Department Column for detailed description of the invention in the specification to be amended.

6. Contents of amendment (1) FIA specification 4! R3 line “Cysteinoamine, J
is corrected to "cysteamine."

(2) Same book, same page/1st line r3!20.2♂ OJ [3
Corrected to Ko 30.2930J.

(3) Same book, same page/3rd line “773” as r 7 , r j
Correct to J.

(4) In the 3rd line of page 1 of the same book, "target compound substance" is corrected to "target compound."

(5) Ibid., p. 12, lines 3 to 3, “Benzyl dimethyl chloride.”
"Cetyl ammonium" is corrected to "Benzyl cetyl dimethyl ammonium chloride".

(6) On the same page of the same book /I-/line 2, "active substance standard" is corrected to "active fraction standard."

(7) In lines 7 to 2 from the bottom of page 13 of the same book, "active" is corrected to "reduced form."

that's all

Claims (1)

[Claims] A compound represented by: and a pharmaceutically acceptable salt thereof. A method for producing a compound represented by and a pharmaceutically acceptable salt thereof, which comprises subjecting the compound represented by or a salt thereof to a reduction reaction.