DD272466A5 - STREPTOGRAMINE B METAL COMPLEXES, THEIR PREPARATION AND THEIR USE IN ANIMAL FOOD - Google Patents

Abstract

According to the invention Streptogramin B metal complexes of the general formula a & SB! In which SB a streptogramin B antibiotic Me is a physiologically compatible metal cation such as copper, zinc, manganese, iron, cobalt, magnesium, calcium or aluminum, n2 or 3 anda number from 1 to n, prepared by adding a streptogramin B antibiotic and a physiologically acceptable metal salt act in geloester form each other, neutralized with an inorganic base and isolated from the reaction mixture, the complex according to known work-up methods. The Streptogramin B metal complexes prepared according to the invention are valuable performance enhancers in animal nutrition.

Description

a (Et) x Me ^{n +} x (na) OH

in which SB Etamycin (= Neoviridogrisein IV), characterized in that one allows a Streptogramin B antibiotic and a physiologically acceptable metal salt act in dissolved form on each other, neutralized with an inorganic base and isolated from the reaction mixture, the complex according to known work-up methods.

2. The method according to claim 1, characterized in that "Me" copper, zinc, manganese, iron, cobalt, magnesium, calcium or aluminum means.

3. Feed additive, characterized by a content of a streptogramin B metal complex according to claim 1.

4. Feed additive, characterized by a content of an etamycin-metal complex according to claim 2.

5. feed additive according to claim 3, characterized in that it consists of a mixture of different Streptogramin B metal complexes of the formula II.

6. Use of Streptogramin B metal complexes according to claim 1, characterized in that they are used as an additive to animal feed.

7. Use of etamycin-metal complexes according to claim 2, characterized in that they are used as an additive to animal feed.

Field of application of the invention

The invention relates to a process for the preparation of streptogramin B metal complexes and their use in animal nutrition.

Characteristic of the known state of the art

The streptogramin B-type antibiotics include etamycin (also called viridogrisin or neoviridogrisin IV (NVG IV)), neoviridogrisin I, II and III, and grividomycin I, II and III. They are characterized by the following formula:

CH ₉

CO-NH-CH-CO-NH-CH-CO-N CH-CH,

³

I co

CH-NH-CO-CH-NH-CO-CH

N-CH, I ³ CH ₂

I CO

NO,

CH-R ₂

CH-CH.

\\ CH ₃ I R4 ^CH 3 R H rj CH ₃ VIRIDOGHISEIN CH, etamycin OH CH, CH, CH, NEOVIRIDOGRISEINIV CH, CH, VIRICOGRISEINII OH CH ₃ CH ₃ CH, neoviridogriseini H CH ₃ C ₂ H ₆ CH ₃ neoviridogriseinii H CH ₃ CH ₃ neoviridogriseiniii OH C ₂ H ₈ H GRIVIDOMYCIN l / ll OH H GRIVIDOMYCINIII OH CH ₃

(Ref: Vazquez, Antibiotics III, Mechanism of action of antimicrobial and antitumcr agents, p. 521 et seq. (11) 75]).

They are formed - usually as a mixture of several antibiotics - in the fermentation of certain Streptomyces species. So

is produced by cultivating Streptomyces griseus NRRL 2426 etamycin in addition to griseoviridine (US Pat. No. 3,023,204) and '

Streptomyces sp. P 8648 (Form - P 3562) in addition to etamycin the NVGs WII; (U.S. Patent 4,536,397). Finally, according to the

European application 87101796.8 in the fermentation of Streptomyces sp. Y-8240155 (DSM 3840) except etamycin, among others still the Grividomycine I, II and III formed).

The streptogramin B antibiotics are effective against Gram-positive bacteria and mycoplasmas. Application in

in veterinary medicine and as feed additives for improving weight gain in farm animals (US Pat. Nos. 4,355,122 and US Pat

Object of the invention

The aim of the invention is the provision of new Streptogramin B antibiotics for use in animal nutrition, which have a nutrient utilization enhancing and thus growth-increasing effect.

Explanation of the essence of the invention

The invention has for its object to find new Streptogramin B antibiotics with the desired properties and processes for their preparation.

It has now been found that metal complexes of said compounds mixed with feed already in very small quantities lead to a highly significant and uniform weight gain of the animals, the higher ice is achieved when administering the free antibiotics.

The present invention thus metal complexes of streptogramin B-Antlblotlka, their preparation and their use as growth-enhancing and nutrient utilization enhancing agents in Tlerernfihrung. The metal complexes contain as central atoms physiologically acceptable divalent or trivalent cations of metals which are generally used as trace elements in animal nutrition, such as copper, zinc, manganese, iron, cobalt, magnesium, calcium and aluminum. The ratio NVQ: Metallkatlon is between 1: 1 and (depending on the valency of the cation) 2: 1 or 3: 1. So the general formula of the complexes is

a | SB) xMe ^nt x (na) OH Il

where SB is a streptogramin B antibiotic, Me is the metal cation, η is 2 or 3, and a is a number from 1 to η (η 2 a). Within the specified range, the composition of the complexes can vary as desired. It is therefore possible to choose the composition which appears most advantageous and suitable for the purpose of the application. The complexes of etamycin are preferably the antibiotics of the formula I, since this forms the main product in the fermentation of said streptomycetes. Also suitable are mixtures of complexes of two or more metals which may be stoichiometrically different in composition, e.g. Mixtures of the complexes of copper / zinc, manganese / cobalt, magnesium / iron, copper / manganese / magnesium, etc. Mixtures of complexes of different Streptogramin B antibiotics are also of practical importance. This offers the advantage that one can complex the obtained during the fermentation antibiotic mixtures without prior separation.

The complexes are prepared from the antibiotics of formula I and metal salts in solution. The former are soluble in pure form and as mixtures in many organic solvents such as, for example, methanol, ethanol, propanol, isopropanol, butanol, methylene chloride, ethyl ether, tetrahydrofuran, dioxane, dimethylformamide, ethyl acetate and ether. The solution of the metal-free antibiotics (pure or as a component mixture) is mixed with shaking or stirring with the amount of metal salt solution necessary for the desired composition; then an inorganic or organic base equivalent to the amount of metal salt is added.

The metal salts are derived from any physiologically compatible inorganic and organic acids. Suitable z. Chlorides, bromides, sulfates, phosphates and carboxylates of organic acids, e.g. Formates, acetates, sulfonates, phenates and enolates. The salts can be used both as hydrates and in anhydrous form. For implementation, they are at room temperature in water or a polar organic solvent such. As methanol or dissolved in mixtures of organic solvents with water.

The operations are preferably carried out at room temperature, but may also be carried out above and below, if this requires the solubility of the components or the volatility of the solvents. Complex formation is very rapid and occurs with the use of colored metal ions, e.g. Cu (II), Co (IIl or Fe (III) ions can also be recognized by a momentary change in color or a change in the UV and visible spectrum, and the addition of a base subsequently neutralizes the protons released during the formation of the complex. For this purpose, all organic and inorganic bases which do not strongly complex with the metal ions themselves may be used, Among the inorganic bases, caustic soda and potassium hydroxide are particularly suitable., Alcoholates such as sodium methylate or organic bases may also be used with the same success. Bases white triethylamine, triethylamine, N-methylpiperidine, pyridine, collidine and diisopropyl-ethylamine can be used.

The complexes are isolated by concentrating the reaction solution at room temperature in vacuo or at atmospheric pressure. From the residue, uncomplexed salts and water-soluble impurities are washed out with water. The filtered metal complexes may optionally after drying with less polar organic solvents. getting washed. This will allow you to remove non-complexed portions of Neoviridogriseine or any organic contaminants present. Suitable organic solvents are ethers such as diethyl, di-i-propyl, dipropyl and dibutyl ether.

The metal complexes of the invention are surprisingly stable compounds. They do not change in the air or in daylight and are not hygroscopic. In water, they are difficult to dissolve. They dissolve in contrast to the free antibiotics also difficult in little polar or apolar organic solvents wio ether; lighter in alcohols. They have higher decomposition points and can be more thermally stressed than the uncomplexed antibiotics. Due to their physical properties, they can be well distributed and mixed with other feed materials. Because of their antibiotic properties, very good compatibility and their physiochemical properties, the complexes according to the invention are very suitable as growth promoters in animal nutrition, especially in the feeding and rearing of poultry (chickens, laying hens, turkeys), piglets and fattening pigs and cattle and lambs. They may be co-administered with any other feed additive (e.g., antimicrobial agents, other antibiotics, inert fillers such as potassium carbonate or silica gel, and animal feed of various compositions). Particular mention may be made of mixtures of the complexes with their underlying complex-free antibiotics, which can be produced either by simple mixing or by applying the metal salt in the stoichiometric deficit during complexation.

A preferred application form is the addition to the feed in the form of a concentrate (Praemix). The concentrate can be prepared, for example, by mixing the active ingredient, the crude product or the active ingredient-containing mycelium with a physiologically compatible, solid or liquid carrier. As a solid carrier, for example, grain by-products such as wheat, wheat bran or de-oiled rice bran, but also maize meal, soybean meal, bolus alba or calcium carbonate and cellulose derivatives such as methyl and carboxymethyl be considered. As liquid lazy; For example, physiological salt solutions, distilled water and physiologically compatible organic solvents can be used. It is also possible to add suitable additives such as emulsifiers, dispersants, suspending agents, wetting agents or gelling agents. These concentrates may generally contain about 0.5 to about 5% of the active ingredient, and depending on the application, the active ingredient concentration can also be significantly exceeded or fallen below.

When administered with animal feeds, it is expedient to proceed by mixing a concentrate with the feed, for example by mixing, grinding, pouring and stirring.

\ Another form of administration is that the active ingredient as such or in the form of a concentrate or

suspendible powder is added to the drinking water or other beverage, such as milk substitute food. Depending on the indication and the mode of administration, the dosage may be about 0.02 to about 5.0, preferably about 0.2 to about 2.0 mg of active ingredient per kg of body weight and day. The diet results in concentrations of about O ₁ S-SOO, preferably about 2-100 g drug / ton.

AusfOhrungsbelsplel The invention is explained in more detail below with reference to some examples.

1) Etamycin-copper-IID-complexlEt) χ Cu (II) OH

A solution of 170.5 mg (1 mmol) of copper (II) chloride dihydrate in 5 ml of methanol is added to 879 mg (1 mmol) of etamycin dissolved in 10 ml of methanol. While stirring, 2 mM of methanolic NaOH are added dropwise. The methanol is distilled off in vacuo, the precipitate is rubbed with water, filtered off with suction and washed with water until the filtrate is free of Cl ions. Subsequently, the copper complex,; suspended in ether. The mixture is stirred for 30 minutes, filtered off with suction, washed with ether and dried in vacuo over P ₂ O ₁ . Yield: 907 mg of etamycin copper complex. C ₄₄ Hg] CuNeO ₁₂ (958.54)

boron. C 54.0% H 6.6% Cu 6.5%

gef. 54.8% 6.7% 6.0%

The calculated analytical values are based on a water content (determined according to Karl Fisc!.: R) of 2.0%. A _m ". (CH ₃ OH) = 340 nm

2) Etamycin copper dD complex fEt, x CU (III)

To a solution of 2.64 g (3 mmol) of etamycin in 30 ml of methanol is added a solution of 256 mg (1.5 mmol) of copper (l) chloride.

Added dlhydrate in 15 ml of methanol. The procedure is continued in Example 1 to obtain 2.51 g of etamycin copper complex.

C _M H _m CuN _ie OjJ (1819.53)

C 55.8% H 6.9% Cu 3.4%

gef. 55.5% 6.9% 3.6%

The calculated analysis values are based on a water content (determined according to Karl Fischer) of 4.0%. A _n ,.,. (CH ₃ OH) = 353 nm

3) Etamycin cobalt dD complex Et x Co (II) OH

4.40 g (5 mmol) of etamycin and 1.21 g (SmMoI) of cobalt D-chloride hexahydrate (98%) are dissolved in 75 ml of methanol and 10 ml of 1N-methanolic NaOH are added. The procedure is as described in Example 1 and receives 4.6 g of etamycin-cobalt complex.

Co. 5.6% (based on 7.0% water content, which after

gef. 6.6% Karl Fischer was determined)

On _1x (CH ₃ OH) = 347 nm

4) Etamycin-Zn (II) complex (Et ₂ χ Zn (II))

Solutions of 4.40 g (5 mmol) of etamycin in 50 mL of methanol and 348 mg (2.5 mmol) of zinc chloride (98%) in 25 mL of methanol

are combined and mixed with 5 ml of 1 N methanolic NaOH with stirring. The procedure is as described in Example 1 and receives 4.65 g of etamycin-zinc complex.

C ₈₈ Hi ₂₂ Nt ₆ O ₂₂ Zn (1821,35)

C 53.8% H 7.1% Zn 3.3%

gef. 54.3% 7.1% 3.3%

The calculated analysis values are based on a water content (determined by Karl Fischer) of 7.3%. A _{m <x} . (CH ₃ OH) = 350 nm

5) Etamycin iron complex Et ₃ x Fe (III)

Dissolve 5.27 g (6 mmol) of etamycin in 60 ml of methanol, add the solution of 546 mg (2 mmol) of iron (III) chloride hexahydrate (99%).

in 30 ml of methanol added and added with stirring with 6 ml of 1 N methanolic NaOH. Work-up as in Example 1. 5.4 g of etamycin-iron complex are obtained.

Ci ₃₂ H ₁₈₃ FeN ₂₄ O ₃₃ (2689.82)

C 57.2% H 7.0% Fe 2.0%

gef. 57.5% 7.1% 2.1%

The calculated analytical values are based on a water content (determined according to Karl Fischer) of 3.0%. A _m "i (CH ₃ OH) = 304 nm AWi (CH ₃ OH) = 340nm (shoulder)

6) Etamycln-aluminum complex-t ₃ x .1 (111)

A solution of 483 mg (2 mmol) of aluminum chloride hexahydrate in 30 ml of methanol is added to a solution of 6.27 (6 mmol). Etamycln Added to 60 ml of methanol. After addition of β ml of 1 N-methanolic NaOH is worked up according to Example 1. This gives 5.35 g Etamycln-Alumlnlkomplex.

C ₁₃₂ H ₁₁₃ AIN ₂₄ O ₃₃ (2660.96)

calc. C 57.5% H 7.1% Al 1.0%

gef. 57.3% 7.2% 1.0 · «.

The calculated analytical values are based on a water content (determined according to Karl Fischer) of 3.5%.

λη,., 1 (CH ₃ OH) - 314nm

AmM ₃ (CHjOH) - 334 nm (shoulder)

7) Etamycin calcium complex Et] χ Ca (II)

4.40 g (5 mmol) of etamycin and 553 mg (2.5 mmol) of calcium chloride hexahydrate (99% Ig) are dissolved in 75 ml of methanol. 5 ml of 1 N-methanolic NaOH are added and the reaction is carried out as described in Example 1. This gives 2.9 g of etamycin calcium complex.

C ₈ H ₁₂₂ CaNuO ₂₂ (1796.06)

C 56.0% H 7.1% Ca 2.1%

gef. 56.5% 7.3% 2.0%

The calculated analysis values are based on a water content (determined according to Karl Fischer) of 4.9%. A _ffl i (CH ₃ OH) = 304 nm

λη,., 2 (CH ₃ OH) = 345 nm (shoulder)

8) Etamycin magnesium complex Et ₂ χ Mg (II)

The solution of 8.79 g (10 mmol) of etamycin and 1.02 g (5 mmol) of magnesium chloride hexahydrate in 15OmI of methanol is added under stirring with 1Om11 n-methanolic NaOH. The mixture is worked up by the method of Example 1. 6.4 g of etamycin-magnesium complex are obtained.

C 56.3% H 7.1% Mg 1.3%

gef. 56.4% 7.2% 1.3%

The calculated analysis values are based on a water content (determined according to Karl Fischer) of 5.2%. A _m ", (CH ₃ OH) = 312 nm A _m " ₂ (CH ₃ OH) = 343 nm

9) Etanycine magnesium complex Et χ MgOH

1.76 g (2 mmol) of etamycin dissolved in 20 mmol of methanol are added successively with a solution of 407 mg (2 mmol)

Magnesium chloride hexahydrate in 16 ml of methanol and with 4 ml of 1 N methanolic NaOH. After that described in Example 1 Method gives 1.66g Etamycin magnesium complex.

C ₄₄ H ₆₂ MgN ₈ O ₁₂ (919.31)

Calc. Mg 2.5% (based on 4.9% water content, the after

gef. 2.6% Karl Fischer was determined)

A _m "i (CH ₃ OH) = 312 nm A _m "j (CH ₃ OH) = 344 nm

10) Etamycin manganese complex Et ₂ χ Mn (II)

A solution of 3.52 g (4 mmol) of etamycin and 400 mg (2 mmol) of manganese (II) chloride tetrahydrate in 60 ml of methanol is mixed with 4 ml of 1 N methanolic NaOH. The method described in Example 1 gives 3.4 g of etamycin-manganese complex.

C 55.1% H 7.0% Mn 2.9%

gef. 55.7% 7.1% 2.9%

The calculated analysis values are based on a water content (determined according to Karl Fischer) of 5.6%. Am (CH ₃ OH) = 350 nm

Biological examples Example I: A feeding trial with etamycin-Cu complex according to Preparation Example 2 was carried out with 120 male broilers in

3 groups of 40 chicks each kept in heated batteries (5 chicks at 8 reps).

The following preparations and dosages were used:

1st group: untreated control

2nd group: Etamycin-Cu complex

3rd group: Etamycin-Cu complex

mg / kg feed ppm

6 20

The forage had a commercial composition with 13.0 MJ / kg »3100 kcal / kg and a crude protein content of 22%, with a standardized amino acid, vitamin, mineral · and trace element content.

Composition in%

Fishmeal 6.00

Feed yeast 2.00 Fat (beef tallow) 4.70 Soybean meal 24.00 Lucerne green flour 1.00

Corn 44.89

Wheat 6.35

Wheat flour 8.50

phosphors. Feed lime 1.40

carbonate. Feed lime 0.06

Trace element mixture 0.04

Livestock salt 0.09

Methionine DL 0.07

The trial ran for 6 weeks. Food and drinking water were offered at will. The animals were weighed individually weekly, the live mass (LM) additions were determined, the feed intake was determined and from feed intake: LM increase in feed expenditure was calculated. The mortality was recorded and from all these parameters the production number (PZ) was calculated.

tflgt. LM increase (g) χ survival rate (%)

Feeding expenses x 10 LM increase % Rel. feed intake % Rel. Example I ing ing (6 weeks) 100 100 dose 1320 109.C 2531 '101.2 mg / kg feed 1439 · 109.2 2561 104.5 ppm 1442 · mortality 2644 control - % Rel. % production number Etamycin-Cu 5 10 complex 20 100 20 , 147 feed 92.7 7.5 154 expenditure 95.3 173 control 1.92 Etamycin-Cu 1.78 complex 1.83

'P 0.05 (P - probability of error)

Example II: In a fattening-chick experiment lasting 42 days, rt.it was given 144 male broilers (Lohmann), 3 χ 48 chicks

(8 repeats × 6 animals), the influence of etamycin-Cu complex according to Example I in dosages of 5 and 10 ppm compared to negative controls without added tested. Posture and feeding were the same as in example

The following groups were used:

1st group: untreated control

2nd group: Etamycin-Cu complex

3rd group: Etamycin-Cu complex

mg / kg feed (ppm)

5 10

dose LM increase % Rel. - 7 - 272 466 % Rel. production number mg / kg feed ing feed intake ppm 100 ing 100 173 - 1559 109.0 106.3 207 control 6 1699 » 107.3 3068 101.0 202 Etamycln Cu 10 1673 · mortality 3260 complex feed % Rel. % 3099 expenditure 8.3 1.97 100 2.1 control 1.92 97.5 6.3 Etamycln-Cu 1.85 93.9 complex

• PO ₁ OS

Example III: In another fattening chick experiment, the influence of etamycin-Zn complex according to Example 4 in a dosage of

10ppm compared to a negative control as in Example I tested. 36 broiler chicks (6 repeats χ 6 animals) were used, with a trial period of 6 weeks. Housing and feeding conditions were the same as in Example I.

The following groups were used:

mg / kg feed (ppm) % Rel. feed intake % Rel. 1st group: untreated control - ing 2nd group: etamycin-Zn complex 10 100 100 dose LM increase 109.4 2966 102.5 mg / kg feed ing mortality 3041 ppm % production number Control - 1698 2.8 Etamycin-Zn-10 1748 · 6.6 199 feed % Rel. 228 expenditure Control 1.86 100 Etamycin-Zn-1.74 93.5 complex

Example IV:

The influence of the etamycin-Mn complex according to Example 10 in doses of 5 and 10 ppm in 3 groups of 32 broilers (4 repeats x 8 animals) over 5 weeks was compared with a negative control (without addition) with 96 male broilers ) checked. The husbandry and feeding conditions as well as the evaluation were the same as in example I.

The following groups were used:

untreated control mg / kg feed (ppm) % Rel. feed intake % Rel. I.Gruppe: Etamycin-Mn complex - ing 2nd group: Etamycin-Mn complex 5 100 100 3rd group: 10 • 106.4 2334 100 Results dose • 108.3 2334 103.4 mg / kg feed mortality 2413 ppm % production number - 6.3 control 5 0 204 Etamycin-Mn 10 LM increase 0 222 complex feed ing 222 expenditure 1.78 1310 control 1.67 1394 Etamycin-Mn 1.70 1419 complex % Rel. 100 93.8 95.5

Claims (1)

1. A process for the preparation of streptogramin B metal complexes of the general formula a (SB) x Me ^{n +} x (na) OH II where SB is a streptogramin B antibiotic Me a physiologically compatible metal cation η 2 or 3 and a is a number from 1 to η
means, or
metal complexes