DE1134071C2 - PROCESS FOR THE PRODUCTION OF NEW DERIVATIVES OF TETRACYCLIN-ANTIBIOTICA - Google Patents

Description

2 moles of formaldehyde and about 1 mole of lysine are converted. The products thus obtained are at parenteral and oral application effective.

The reaction between the three components mentioned above can be carried out in a one-step process by reacting all three components with one another at the same time. Man but can also proceed in such a way that the formaldehyde is reacted first with one of the other two components and then the remaining component ——- ^ adds.

Tetracycline in the form of the free base is preferably used as the tetracycline antibiotics. Dei Formaldehyde is preferably used in the form of aqueous or alcoholic solutions. Ar Instead of formaldehyde, one can also use compounds which formaldehyde under the reaction the ability to create conditions.

The reaction is carried out in the presence of solvents or diluents Organic solvents such as

The present invention relates to a methanol, or mixtures of water and organic drive for the production of new derivatives of the 55 solvents used. Water alone cannot be used aii tetracyeline antibiotics, which are readily soluble in water solvents. Are, besides, a nearly neutral _pH value yield the A-Stufcn reaction is conveniently carried out at 6kei and by the oral, intravenous and intramuscular arrival _eiwas elevated temperature, eg. B. 30 to 40 ° C, are effective by turning. led, wohei methanol or mixtures of methano

The term "Tetracyelin antibiotics" should, if ^6o and a little water are used as solvents used in the present specification, _{if you ar} behet _{in 2wei steps,} you can at Chlortetracychn, Oxytetracyciin, tetracycline, Des spielswcise'Lysin 1 MoI formaldehyde in wäßrig® methyltetracychn and such derivatives of this compound _or alcoholic solution or fertilize with 2 mol form include that are antibiotic. convert aldchyd in aqueous solution. So formed.

It is known that derivatives which are easily soluble in water ° ⁵ intermediate can be isolated from the solution, ii of the tetracycline can be obtained if tetra-methanol and a little water are dissolved and then to de cyclm with formaldehyde and certain amino solution of the tetracycline in Methanol at a Tempe connections, z. B. a dialkylamine, piperidine, _ratU r from about 30 to 40 ⁰ C are given.

When tetracycline and formaldehyde are reacted in the first stage, the reaction becomes convenient carried out by suspending tetracycline in a suitable diluent such as butanol, adding the formaldehyde at normal temperature and the mixture thus obtained is heated to boiling under reflux for about 30 minutes. The reaction product will precipitated and isolated by cooling. The isolated product and the equivalent amount of lysine are in "In" a suitable solvent such as methanol, dissolved, and as mentioned above, the solution is heated to a slightly elevated temperature

The chemical constitution of the reaction products has not yet been fully elucidated. Probably the reaction product is a lysinomethyltetracycline of the general formula

A —CH, -B,

wherein A is the remainder of a tetracycline antibiotic and B represent the remainder of lysine.

The analysis confirms the correctness of this assumption. The optical rotation of the reaction products is different from the values of the corresponding tetracycline anribiotic or lysine. The reaction

product is relatively stable to water. The aqueous solutions of the reaction products remain clear for about 24 hours at room temperature. This enables the reaction products to pass through Isolate freeze-drying of their aqueous solutions.

Regarding the therapeutic properties of the new products, the following was found:

The blood levels of patients who, in part, underwent a treatment with the prepared according to the invention L-lysinomethyltetracycline, partly with pyrrolidinomethyl-

i _S tetracycline (a product according to the German Auslegeschrift 1044806) and partly subjected to tetracycline hydrochloride are shown in Fig. 1. The products were administered intravenously. From Fig. 1 it can be seen that according to the invention

ao manufactured product leads to higher blood levels than the other two products. Each patient received 100 mg (calculated on a tetracycline base). This dose was administered all at once in each case.

Fig.

jugjmi M

0.5

L-lysinomethyltetracycline Pyrrolidinomethyl tetracycline Tetracycline hydrochloride

Microbiological determination on agar plate with Bacillus cei'eus varietas mycoides ATCC 9634

\ t -

hours

1 2

12th

From Fig. 2 it can be seen that this is the case when the products are administered intramuscularly. Invention manufactured product at one observation- 65 Each patient received 250 mg (calculated on TetracycHn-

duration up to 24 hours also higher blood levels gives base). In each case, this dose was taken all at once.

as pyrrolidinomethyltetracycline (a product delivered speaking of the German interpretative document 1044806),

Fig.

■ L'Lysinomcthyltetracydin PyrrolidinoracthyltetracycHn

Microbiological determination on agar plate with Bacillus, cereus varietas mycoides ATCC 9634

hours

/ 2

FIG. 3 shows the blood levels that result with the L-lysinomethyltetracycline produced according to the invention and tetracycline hydrochloride when the products are administered orally. Every patient received 150 mg (based on tetracycline) every 6 hours.

Fig. 3 shows the blood levels after 6, 12, 30 and 54 hours were obtained. The blood levels obtained with the product manufactured according to the invention were higher than the blood levels obtained with tetracycline hydrochloride.

3-

Fig.

L-Lysinomethyltetracycline Tetracycline Hydrochloride

Microbiological determination on agar plate with Bacillus cereus varietas mycoides ATCC 9634

Hours 54

Table 4 shows the toxicity towards mice of tetracycline hyiirochloride, pyrrolidinomethyltetracycline (a product produced according to the method of German Auslegeschrift 1044806) and L-lysinomethyltetracycline (a product produced according to the invention). The expression DL ₅₀ means the acute toxicity in mg / kg. The products were administered intravenously. The volume of each dose was 0.4-0.6 ml / 20 g. The doses were administered at a rate of 1 ml / minute. From Table 4 it can be seen that the toxicity of the product prepared according to the invention is considerably lower than the toxicities of the other two products.

soluble yellow powder obtained; it melts above 200 ^° C. with slow decomposition. Yield 5.8 g = 85 _^0/0 of the theory.

Analysis for Calculated found

C 57.79%, H 6.35%, N 9.25%; C 57.59%, H 6.82%, N 8.52%.

Table 4

DL ₆₀

DL _U , calculated on tetracycline hydrochloride

Tetracycline hydrochloride

220

220

Pyrrolidinomethyl tetracycline

125

115

L-lysino

methyl-

tetracycline

320

253

From Figs. 1 to 3 and Table 4, the possibility of the dose of L-lysenomethyltetracycline results to reduce in order to obtain the same blood levels and the same therapeutic effects as with Pyrrolidinomethyltetracycline or Tetracycline Hydrochloride are available.

example 1

5 g of tetracycline base are dissolved in 100 ml of methanol. 2.25 g of L-lysine hydrochloride in 7 ml of water and 1 ml of 38% formaldehyde are added to this solution.

The solution is left to stand at room temperature for 3 hours and then concentrated almost to dryness in vacuo at a low temperature. The residue is dissolved in 25 ml of distilled water and lyophilized. It is finally dried over P, O _{S in a vacuum.}

The light yellow substance thus obtained melts at 190 until 195C. Yield 6.8 g = 94.5 · /, of theory.

Analysis for C _M H _M N, O _n -HCl:

Calculated ... C 54.49 · / ,. H 5.99%, N 8.76%; found ... C 55.18%. H 6.71%, N 8.58%.

Moisture 7.78% (determined according to the Karl Fischer method).

The product can be converted into an acidic addition salt- * ίο <z. B. with inorganic acids such as hydrochloric acid, sulfuric acid, or with organic acids such as formic, acetic, oxalic, citric and ascorbic acid). The addition salts can be formed by dissolving the product in water and adding the equivalent amount of acid.

Example 3

2.4 ml of 38% aqueous formaldehyde are added to a solution of 6 g of tetracycline in 200 ml of methanol given at a temperature of 30 C.

2 g of L-lysine are dissolved in a mixture of 3.4 ml of water and 12 ml of methanol and added to the first solution. The process is carried out analogously to Example 2. The product is a yellow powder which is soluble in water and slowly begins to decompose above 190.degree. Yield 6g = 74% of theory. {%] $ ~ -195 ° (0.5% water).

Example 4

To a solution of 5 g of tetracycline base in 200 ml 1.78 g of L-hydroxymethyl lysine, dissolved in methanol, are slowly added with stirring at room temperature 1.5 ml of water and 2 ml of methanol are added.

The precipitate is filtered off after 90 minutes, washed with ether and dried ^{at 50 ° C. in vacuo.}

The product obtained in this way is a yellow powder that slowly decomposes ^{above 200 ° C.} Yield 5 g = 73.7% of theory,

Analysis for

Calculated found

Moisture: 6.7%.

C 57.79%, H 6.35%, N 9.25%; C 57.12%, H 6.33%, N 9.44%.

The L-hydroxymethyllysine can be obtained as follows be: I mole of L-lysine is dissolved in a small amount of white methanol. To the thus obtained Losing a solution of I moles of formaldehyde ia Methanol added. The reaction product is

SS precipitates, decelerates with a small amount of methanol washed.

Example p Example 2

I ml of 38% aqueous formaldehyde is converted into a solution of 5 g of tetracycline base "" 160 ml of methanol given: 1.75 g of nt-lysine is added at 30 ° C. while stirring ia 3 ml of water and 10 ml of methanol are added. To A precipitate forms a few minutes. Stirring is continued for about 2 hours. Of the Precipitation is removed and with a slight Amount of methanol-ether washed. After drying in vacuo at 60.degree. C. it becomes very light in water I ml of 38% aqueous formaldehyde becomes one Added a solution of 5 g of tetracycline in 160 ml of methanol. While stirring at 3OC, 1.75 g of L-Lysmhase in 3 ml of water and 10 ml of methanol are added. Precipitation begins after a few minutes. That Stirring continued for 2 hours. The precipitation is filtered off and washed with a small amount of methanol-ether. After drying in vacuo at 60 C it becomes yellow, easily soluble in water Polver received; it melts above 200X eater

509608/375

slow decomposition. The yield is 82%; ry] »200 ° (0.5% water).

Analysis for C ₂ , H _3l) N ₄ O, ₀ :

Calculated ... C 57.79%, H 6.35%, N 9.25%;
found ... C 57.30%. H 5.95%, N 9.02%.

Moisture: 10.49%.

Example 6

1.1 ml of 35% strength alcoholic formaldehyde au a solution of 5 g of tetracycline in 200 ml of methanol given. 1.65 g of i.-lysine are added at 35 ° C. with stirring Dissolved in a mixture of 5 ml of water and 7 ml of methanol and added to the first solution. The suspension is stirred for 90 minutes and then filtered ^ The precipitate is washed with a small amount of ether washed. The yield is 5.2 g of a yellow powder which begins to set above 200 ° C.

Corresponding products were made from Oxytetracycline, Dcsmethylchlortetracycline and Desmethyltetracycline produced when the above-mentioned antibiotics are used in place of the tetracycline used in Example 6 became.

The chemico-physical properties of the products obtained are as follows:

Oxytetracycline derivative

Decomposes slowly at 24O ⁰ C
[ft]? = -100 ° ± 4 (0.5% in water)

Desmethylchlorotetracycline derivative
Slowly decomposes at 190 ° C
{<*]? = -242 ° ± 4 (0.5% in water)

Desmethyltetracydine derivative
Decomposes slowly at 180 ° C
[ _Λ ] »= —238 ° ± 4 (0.5% in water)

Example 7

10.5 g of tetracycline base are suspended in 160 ml of tertiary butanol: to the mixture thus obtained 4.1 ml of 38% aqueous formaldehyde and a few crystals of sodium carbonate are under Stirring added.

The mixture is stirred for 15 minutes at room temperature, then heated to the boil and refluxed for 30 minutes. The solution is filtered while hot to remove traces of insoluble S material. The filtrate is cooled to room temperature, the precipitate thus obtained is filtered off and dried ^{at 50 ° C. in vacuo.}

The product is a light yellow powder that is easily soluble in water and will decompose above 190 ° C begins.

To a solution of 2 g of this substance in 60 ml of methanol, a solution of 0.618 g of i.-lysine in Added 1 ml of water and 4 ml of methanol.

Precipitation soon begins; stirring is continued for 1 hour. The precipitate is filtered off and dried ^{at 50 ° C. in vacuo.} The product is a yellow powder which is readily soluble in water and has the same characteristic features as the product described in Example 5. Yield 1.6 g so = 59% of theory. {%) ',> 200 ° (0.5% water).

Analysis for C ₂₉ H ₃₈ N ₄ O ₁₀ : H 6.35%. N 9.25 ° / Calculated ... C 57.79%. H 6.67%, N 9.47 ° / found ... C 57.42%, , ₅ moisture : 5.47%.

Example 8

0.55 ml of 38% aqueous formaldehyde and a solution of 1.15 g of L-lysine hydrochloride in 3 ml of water are added to a solution of 3 g of Chlortet raeyclin in 150 ml of methanol. The mixture is stirred for 4 hours at room temperature. The precipitate is filtered off and washed with methanol-ether. The product obtained after drying in vacuo at 60 C, a yellow powder, which begins to decompose above 190 ⁰ C. Yield 3.23 g = 80% of theory _v.

Analysis for C "H ₃₇ ClN ₄ O ₁₀ · HCl:

Calculated ... C 51.70%, H 5.68%, N 8.31%;

found ... C 52.04%, H 6.07%, N 7.58%.
Moisture: 0.99%.

Claims (3)

Morphoün or piperazine, converts the aminomethyl ·. group, which is thereby stirred into the tetracycline molecule, came to a carbon atom of the tetracycline ring system (German Auslegeschriften 5 1044806 and 1063598) or to the nitrogen atom of the carboxamide group of the tetracycline molecule (bri-Palentanspriiche: multiple patent specification 809585 and German patent 1088481 The aminomethyl derivatives obtained in this way are just as effective antibacterially as tetracycline when they are administered intravenously; t ends; however, they are much less effective than tetra- 1. Process for the production of new deriva- cydin, if it is used orally (E. N. E wald and ten of the tetracyeline antibiotics by implementation G. S. Red in. Antibiotics and Chemotherapy, I960, of 1 mole of a Tetiacycün aatibiotic with 1 to s. 419 to 421). Mol of formaldehyde and 1 mole of a third reactive »5 A number of amino acids are also not capable of ionization, an amino group-containing grain capable, with TetracycÜB antibiotic and formaldehyde components at temperatures of 30 to 40 ⁰ C in with the formation of water-soluble products that have a presence Solvents, such as methanol, have sufficient stability towards hydrolysis, characterized in that ^{reactive ao} _Z u containing an amino group can be used _{as the third vn d for parenteral purposes.} For example, the reaction component lysine is used. of 1 mole of tetracycline, 1 mole of formaldehyde and 1 mole 2. The method according to claim 1, characterized in that the glycocoll obtained product in aqueous solution is stable as a tetracycline antibiotic that the poorly soluble tetracycline is used as L-lysine after a short tetracycline and lysine. If you add ornithine or arginine to the. ^a 5 place of glycocolla used, PnSducts are 3. The method according to claim 1, characterized in that it also has poor stability indicates that the reaction is carried out in two stages by water. is performed by either the tetracycline £ _s has now been found that new and better understood Anübioticum or lysine initially with formaldehyde-bearing derivatives of the tetracyeline antibiotics, which are converted on their own and only the resulting 3 «> _{eme have} better water solubility and good stability is brought. \ Mole of a tetracycline antibiotic with about 1 to