DE1768971C - 11 a-H alogen-e-deoxy-e-desmethyl-emethylene tetracyclines and their acid addition salts - Google Patents

Description

in which E is hydrogen or a hydroxyl group, X is fluorine, chlorine or bromine, Z is hydrogen or is chlorine, but is only hydrogen if X is fluorine or E is a hydroxyl group, and their acid addition salts.

Features. As a numbering system, the d »Chemical Abstracts« used.

OH

OH

Among the biologically active members of this group are those soft the following remainder contain:

Substitutes

common name

The invention relates to new 1 la-halogen-6-deoxyö-desmethyl-o-methylene tetracyclines the general formula

N (CH ₃ J ₂

(Hl)

CONH,

35

OH

in which E is hydrogen or a hydroxyl group, X is fluorine, chlorine or bromine, Z is hydrogen or chlorine, but is only hydrogen when X is fluorine or E represents a hydroxyl group and their acid addition salts. They can be made by using a tetracycline compound of the general formula «

N (CH ₃ ) ₂ 4-N (CHj) ₂ , 6-OH, 6-CH3, 12a-OH 4-N (CHj) ₂ , 5-OH, 6-OH, 6-CH3, 12a-OH 4- N (CHj) ₂ , 6-OH, 6-CH3, 7-Cl, 12a-OH 4-N (CH ₃ I ₂ , 5-OH, 6-CH3, 12a-OH 4-N (CH ₃ J ₂ , 6-CH3, 12a-OH
4-N (CH _{3) 2,} 12a-OH

4-N (CH ₃ J ₂ , 6-OH, 6-CH ₃ , 7-Br, 12a-OH 4-N (CHj) ₂ , 6-OH, 7-Cl, 12a-OH 4-N (CH ₃ ) _2, 6-OH, 12-OH

Tetracycline

5-oxytetracycline

7-ChIortetracydin

6-deoxy-5-oxytetracycline

6-deoxytetracycline

o-Deoxy-ö-desmethyltetracycline

7-bromotetracycline

6-Desmethyl-7-chlorotracycline

o-desmethyl tetracycline

The New Tetracycline Compounds and Procedures their preparation is based on the reaction sequence and structural formulas given below explained. Here, X, Z and E have the above meanings.

The preparation of the new tetracycline compounds is not the subject of the invention.

N (CHj) ₂

OH

CONH,

OH O O O

55

in which E and Z have the above meanings, with a mild halogenating agent known per se in an inert solvent at temperatures converts to about 50'C, the resulting Ha-halo-tetracycline-6,12-hemiketal dehydrated in a manner known per se with a strong acid and optionally from the salt in a manner known per se Way produces the amphoteric compound or other salts.

The tetracycline antibiotics comprise a group of biologically active perhydronaphthacene compounds with the following essential structural OH O

O

Halogenation

N (CH ₃ ) ₂

«• 5

(II)

¹ H ι I II

OH O I OH

CONH ₂

Dehydration

(HI)

CONH,

OH O O 0

ίο

a) lla halogenation

The new 11 a-halogen tetracycUne of the general Formula II graze by reacting a suitable tetracycline of the general formula I with one known mild halogenating agents prepared in a solvent inert to the reaction. The structural formula clearly shows that the halogen atom X in the 11a position of the new Compounds in both cis and trans positions occurs to the hydrogen atom in the 5a pitch of the tetracycline nucleus. It is believed that those compounds in which the halogen atom is in the cis position, predominantly in the specified Structure occur, d. H. in the structure which forms an oxygen bridge between the 6 and 12-position of the tetracycline nucleus, which results from the formation of a hemiketal between the carbonyl group in the 12-position and the hydroxyl group in the 6-position. Shine in a crystalline state these compounds are predominantly in the form of the hemiketal. This «assumption is confirmed by the Absence of carbonyl absorption on ultra-red analysis (no bands in the 5 to 6 micron range) hardened. Of course, such structures exist in equilibrium with the uncatalyzed form of general formula IV

40

(IV)

45

CONH,

OH O O O

This is especially true when such forms are in solution. If in the following of the Hemiketal compounds are spoken of, this term is intended to include both cis-lla-halogen compounds in uncatalyzed form as well as hemiketal. The new epoxy compounds are 11 a - halogen -1,4,4a, 5,5a, 6,11,11 a, 12,12a - decahydro-3,10,12,12a - tetrahydroxy-1,11-dioxo-2-carboxamidonaphthacene-6,12-epoxides. For the sake of simplicity, the new halogen compounds are hereinafter referred to as Called tetracycline-6,12-hemiketalc.

As mentioned, the halogenation by reaction of a suitable tetracycline is the general Formula I carried out with a mild halogenating agent in an inert solvent. Suitable Tetracycline starting compounds are, for example, tetracycline. Oxytctracycline and chlortetracycline. the Tetracycline parent compound must, of course, have a hydroxyl group in the 6-position in order to achieve the To form hemiketal structure. For the production of the aeuen Ila-Halogentetracycline, in which the halogen Is chlorine or bromine, one can use a variety of halogenating agents in one opposite the Implementation use inert solvent. Suitable halogenating agents are, for example, bromine; Chlorine; Iodine chloride; Iodine bromide; low molecular weight alkanoic acid amides which are cbdorinated or brominated on the nitrogen atom, such as N-chloro- and N-bromoacetamide, chlorinated or brominated dicarboximides, such as N-chloro- or N-bromosuccinimide or -phthalimide, or low molecular weight N-alkanoylanilides, such as N-3romacetanilide or propionanilide; 3-chlorine, 3 -Bromo-, 3,5-Dicfclor- and 3,5-Dibromo-5,5-dimethyl-hydantoin; Pyridinium perbromide and perchloride hydrohalides, such as pyridinium perbromide hydrobromide or pyridinium perchloride hydrochloride; and low molecular weight alkyl hypochlorites, such as tertiary butyl hypochlorite. It is evident that in general each one commonly used in the art mild halogenating agents can be used. However, those mentioned above preferred. In general, 1 to 1.2 moles of the halogenating agent are preferably used each Moles of tetracycline starting compound.

An "inert solvent" is understood here to mean a solvent which is defined by the Reaction conditions do not work in an undesirable manner either with the starting compounds or with the end product reacts. A minimum of laboratory tests enables the selection of suitable ones Solvents for the process for the preparation of the compounds according to the invention. Examples such solvents are dioxane, tetrahydrofuran and the methyl ether of diethylene glycol and the methyl ether of ethylene glycol. In general, the connection of water is preferred, when the starting compound is a IIa-bromine or iodine compound, which is somewhat sensitive to Seem to be water. However, this is not essential. If water is present in the reaction mixture is, one should avoid prolonged exposure to the 1 la-bromine compounds in order to avoid a noticeable Prevent decrease in the yield of the desired product. The temperature seems at not to be critical of this implementation. Temperatures from 0 to 50 ° C have proven to be suitable. Temperatures above 50 ° C. are avoided in order to avoid the possible formation of 5a, 6-anhydro compounds to exclude, which reduce the effectiveness of the process. The selection of the best reaction conditions, such as temperature, solvent, halogenating agent, etc., is a matter of routine experimentation. A solvent system is preferably selected from which the Ila-halogen product crystallizes out, especially when it comes to the bromine compounds.

A solvent system made up of benzene and dimethyl ether of ethylene glycol (ratio 1: 1) is a preferred solvent system for IIIa bromination of oxytetracycline. The reaction product separates easily from the reaction mixture. If this product remains in the solution for a long time, there appears to be a noticeable degradation with it a substantial reduction in the product yield. As already mentioned, the Iia bromine compounds appear to be very sensitive to water. Solves

For example, if Ila-bromo-5-hydroxytetracycline-6,12-hemiketal-hydrobromide in water to a solution with a pH value of about 2, then after about I ¹ I ₂ hours the product is no longer detectable because it becomes a Substance that appears to be a lactone has been broken down. If the Ila halogen product is boiled with acetone, the starting compound 5-hydroxytetracycline is recovered in large quantities. On treatment with aqueous sodium hydrosulfite, the starting compound is also obtained from the 1 la-bromo hemiketal. The chloro-hemiketal compounds are generally preferably prepared in a water-miscible solvent, such as tetrahydrofuran, dioxane, acetone, low molecular weight dialkyl ethers of ethylene glycol or propylene glycol. The product is obtained by simply diluting the reaction mixture with water, which precipitates the 11 a-chlorotetracycline-6,12-hemiketal.

The 1 la chlorine compounds are apparently not as highly reactive as the corresponding 11 a-bromine compound, so that special measures are not required during manufacture. These compounds are actually in aqueous solutions quite stable, even at low pH values. For example, 11 shows a - chlorotetracycline - 6,12 - hemiketal hydrochloride if left to stand for 18 hours at a pH of about 1 and room temperature in Water no noticeable decomposition.

The manufacture of the lla-fluorotetracycline takes place by reacting the tetracycline starting compound with perchloryl fluoride in the presence of a strong base, preferably an alkali hydroxide or alcoholate. Implementation is customary carried out so that the starting compound in the selected solvent, preferably a low molecular weight alkanol, which is an alkane with up to 3 carbon atoms, whichever is the least contains a molar equivalent amount of the base, soluble and perchloryl fluoride, which at room temperature is gaseous, admits. As implementation progresses the pH of the solution falls from alkaline values to almost neutral values. The product begins usually deposit at a pH of close to »8. The crystalline product is in u licher way collected and dried.

Besides the 11 a - halo tetracycline - 6,12 - hemi ketalen, the process also leads to lla halogen tetracyclines are compounds in which hemiketal formation has occurred. It is believed, that in these connections the lla-permanent Halogen atom on the other side of the 1 la position Carbon atom stands like the halogen atom of the hemiketals. It is further assumed that the Stereochemical position of the 1 la halogen atom prevents the formation of the hemiketal structure. These compounds are generally obtained by evaporating the reaction mixture after precipitation of the 6,12 hemiketal compounds. The trans-1 Ia halogen compounds are represented by the following general formula in which Z. E and X have the meanings given above:

-OH

CONH ₂

OH O O O

b) dehydration

The 11 a-halogen hemiketals of the general formula II, in which the radical X is a chlorine or fluorine atom, are surprising in substances high in vivo biological activity against disease-causing microorganisms. When tested in vitro, the compounds show noticeable activity. The transformation is obtained by treating the starting compound of the general formula II with a strong acid, which has a dehydrating effect, accomplished. A number of acids are suitable for this purpose. Used with preference mineral acids such as sulfuric acid, phosphoric acid, polyphosphoric acid, perchloric acid, as well as other acids, such as glacial acetic acid (containing boron trifluoride), hydrogen fluoride (preferably in liquid form) and trihaloacetic acids such as trifluoroacetic acid. In addition, the mineral acids are preferred used in concentrated form, for example as at least 60% aqueous acids. If you use Mineral acids, particularly advantageous results are obtained with acids in the following concentrations:

Sulfuric acid 90 to 95%

Phosphoric acid 80 to 85%

Perchloric acid 60 to 70% ⁶ S

Response time and temperature do not appear to be critical. In carrying out this procedure, the starting compound is added to the selected acid and the mixture is allowed to stand for a relatively short time. For example, the starting compound is added to hydrogen fluoride, preferably liquid hydrogen fluoride, at 0 to 50 ^° C. and the mixture is left to stand for 5 to 7 minutes or even longer, for example up to several hours. The hydrogen fluoride is then allowed to evaporate. The residue is treated in the usual way to obtain a crystalline product such as the hydrofluoride salt, for example by stirring in a non-solvent and recrystallization from solvents such as low molecular weight alkanols. If sulfuric acid is used, a temperature below 20 ^° C. is preferably used. Using perchloric acid, so the reaction is preferably carried out at temperatures of about 50 ⁰ C and higher, preferably from about 60 to 70 ⁰ C is performed. More concentrated perchloric acid can be used in this reaction. However, these higher concentrations are not preferred because of the risk of explosion known to those skilled in the art. The reaction time does not appear to be critical as the reaction appears to be virtually instantaneous. Generally, are, for example, a reaction time of 5 to 15 minutes excellent results, if the reaction temperature is 60 to 70 ⁰ C. Longer

Although heating is not disadvantageous, it does not bring any noticeable advantage and becomes from this Basically not done. At lower temperatures, slightly longer conversion times may be necessary be to get good yields. If you use trifluoroacetic acid or acetic acid, so the reaction is preferably carried out at room temperature within about 24 hours. After the conversion has ended, the product is obtained by the usual measures. For example It is very useful to keep the reaction mixture simple Dilute with a nonsolvent, such as water, resulting in precipitation of the product as Acid salt leads. The acid salts obtained can be converted into the free base or into any salt convert in the usual way. If the starting compound for this reaction is an IIIa fluorine compound is used, the best results are obtained using perchloric acid.

The products of the dehydration reaction are 1 la-halo-tetracyclines of the general formula III, which as o-deoxy-o-desmethyl-ö-methylene-lla-halogen tetracycline are designated. The compound in which Z represents a hydrogen atom shows as Mineral acid salt on ultra-red analysis (KBr) cine carbonyl absorption at or near 5.70 microns. It was mentioned above that this absorption in the case of the 11 a-halo-hemiketal starting compound is missing. An interesting shift in the UV absorption maxima (measured in 0.01-n methanolic hydrochloric acid) from the range of 345 ΐτίμ (which is characteristic of the hemiketal starting compounds is) in the range of 375 ιτίμ (which is characteristic of the dehydrated products is) observed. The dehydrated products also show two additional UV absorption maxima, which are about 4.2 or 5 times stronger than the maxima at 375 πΐμ. These two additional Maxima are at or around 240 or 277 πΐμ. The above-mentioned characteristic maximum at about 375 πΐμ in compounds in which Z denotes a hydrogen atom is found in compounds in which Z is a chlorine or bromine atom, at about 380 to 390 ιημ.

The IIIa-halogeno-6-methylene-tetracyclines of the general formula III are valuable starting materials for further substitution reactions, for example for substitution reactions in ring D, ie for nitration and halogenation. Furthermore, in the case of compounds of the general formula III in which E denotes a hydrogen atom, substitution reactions in the 6-methylene group can also take place.

The compounds according to the invention are particularly suitable for the production of valuable 6-methylene tetracyclines by dehalogenation according to the following Reaction scheme;

1 la dehalogenation

Z CH ₂ OH N (CH ₃ J ₂

60 The dehalogenation can be carried out chemically and also catalytically by many of the methods customary for the removal of halogen. Such a catalytic reduction is carried out by treating the starting material with hydrogen gas in the presence of a noble metal catalyst, for example a palladium or rhodium catalyst, at pressures from atmospheric pressure to a pressure of 1000 atmospheres and temperatures from 0 to 100.degree. A variety of chemical methods can be used. A well-known method is the reduction with alkali hydrosulfites. In this case, the starting compound is added to an aqueous solution of the individually selected hydrosulfite and the reaction mixture is left to stand at room temperature, which has been found to be most expedient for this reaction. Further methods of removing the halogen atom in the lla position are:

a) treatment with dilute, aqueous hydriodic acid,

b) treatment with metallic zinc in the presence of a proton donor such as acetic acid,

c) heating the Ila bromine compound in acetone to simmer.

Surprisingly, it has been found that 1 la-halo-tetracyclines can be easily obtained by simultaneous treatment of the 11 a-halogen compound with zinc and sodium iodide in a solvent inert to the reaction (preferably in low molecular weight alkanones such as acetone) at a temperature between room temperature and re-nut temperature using equivalents Can easily dehalogenate up to excess reagent amounts of 1000 mole percent and more. At the reflux temperature, the reaction is complete in as short a time as 30 minutes. After the reaction has ended, the product is obtained by filtration and concentration of the reaction mixture. Under the conditions of reductive removal of the 1 la halogen atom described above, nitro groups are reduced to amino groups, so that the corresponding amino derivatives are obtained.

For the conversion of the 1 la-halogen-6-methylentetracydine i.i the 6-methylene-tetracycline are the 1 la-chloro-o-methylene-tetracycline preferred because this Compounds in general appear to be quite stable and easily dehalogenated. the 1 la-Fluorverbir fertilizers are less easily dehalogenated.

The 1 la-Deha logen compounds show a surprisingly high degree of aa activity when you use them in vitro against a variety of disease-causing

sacherid organisms. They are particularly effective against antibiotic-resistant strains of Microorganisms.

The following Table I summarizes the activity of 6-methylene-tetracycline against a variety of disease-causing agents Microorganisms including antibiotic-resistant strains. The minimum inhibitory concentration is determined by the known technique of serial dilution. The table also shows the corresponding values of 1 Ia-chloro-6-methylene-tetracycline and of 6-deoxytetracycline locked in. It turns out that the values for 6-methylene-tetracycline in general

Table I.

are lower than those for o-deoxy-tetracycline, indicating greater activity, particularly against Micrococcus pyogenes var. aureus400, a tetraeyclin resistant Organism.

1 = o-methylene-tetracycline.

2 = o-deoxy-tetracycline.

3 = 1 la-chloro-δ-methylene-tetracycline.

4 = 6-deoxy-6-desmethyl-6-methyli; n-

5-oxy-tetracycline.

5 = 7-chloro-6-deoxy-6-desmethyl-

o-methylene-S-oxy-tetracycline.

6 = o-deoxy-o-desmethyl-tetracycline.

organism

Micrococcus pyogenes var. Aureus 5

Streptococcus pyogenes
Streptococcus faecalis
Diplococcus pneumoniae
Erysipelothrix rhusiopathiae
Corynebacterium diphtheriae
Listeria monocytogenes
Bacillus subtilis
Lactobacillus casei

Bacterium ammoniagenes

Streptococcus pyogenes 98

Micrococcus pyogenes var. Aureus 209 (p) Aerobacter aerogenes

Escherichia coli

Proteus vulgaris

Pseudomonas aeruginosa

Salmonella gallinarium

Salmonella pullorum

Klebsiella pneumoniae

Neisseria gonorrhoeae

Hemophilus Influenzae

Shigella soniei

Bruceila bronchiseptica

Mallecmyces mallei

Vibrio comma

Fasteurefla multocida Streptococcus agalactiae Mycobacterium 607 Mycobacterium berolinense Candida albicans Sardna lutea

Antibiotic-resistant strains of Micrococcus pyogenes var. Aureus

Line 376 *) Stem 400 **)

*) Tetracycline resistance with a concentrate! «™ below 100 γ IvA. **) Tetracycline resistance at a concentration of outer SO y'm \ 4p / ~ 1 ffHi

Minimum concentration <0.19 3.12 2 0.78 - 6.3 - (r'ml) 50 - - - 6.25 4th 0.78 5 0.25 0.19 1.56 3.12 I. 1.56 6.3 0.19 3 (25 p) > 100 - 25th 1.56 146 <0.19 12.5 0.39 iOO 100 12.5 25th > 100 6.25 0.78 0.03 0.19 100 039 <0.19 25.0 0.78 100 100 25th 3.12 > 100 25th 0.39 0.6 0.19 100 <0.19 3.12 3.12 12.5 25th 100 > 100 12.5 0.39 0.06 0.19 <0.19 0.78 0.39 3.12 63 (5Op) > 100 0.78 0.39 0.125 0.19 0.78 _ 0.78 1.56 3.12 6.3 3.12 12.5 > 100 039 3.12 1.0 0.02 1.56 0.19 <0.19 6.25 0.78 100 (ρ) > 100 0.19 0.19 0.19 0.19 - <0.19 <0.19 0.01 0.09 > 100 108 0.09 0.01 0.04 0.09 0.78 3.12 25th 3.12 12.5 > 100 0.78 0.19 1.56 <0.19 0.19 6.25 50 0.19 <0.19 0.39 0.78 3.12 0.39 0.19 0.19 - <0.19 0.19 0.09 0.01 - - <0.19 039 0.25 0.25 - <0J9 6.3 1.56 <0,! 9 3.12 0.78 <0.19 50 100 3.12 <ai9 12.5 1.56 625 3.15 (p) 0.78 0.78 3.15 (p) 0.78 0.19 0.06 0.19 0.03 3.12 0.39 0.39 0.25 1.56 0.25 0.78 3.12 0.19 04 039 _ _ G, 7S - 100 100 50 100

When repeating these in vitro experiments in the presence similar results are observed for human serum. The following table summarizes the activity of the compounds listed in Table I when tested in 20% human serum:

Table II

Minimum hcmm 5 concentration (y / ml) 0.39 4th Mieroeoccus pyogenes var. 0.78 0.09 a u reu s 5 0.19 Streptococcus pyogenes 0.78 0.39 Streptococcus pyogenes 98 Mieroeoccus pyogenes var. aureus 209 ρ

It should be noted that 7-chloro-6-deoxy-6-desmethyl-6-methylene-5-oxy-tetracycline greater in vitro activity against large numbers of microorganisms as o-deoxy-o-desmethyl-o-methylene-S-oxytetracycline owns.

If o-deoxy-o-desmethyl-o-methylene-S-oxytctracycline is tested in vivo, both orally and parenterally, this compound shows greater activity than tetracycline or 5-oxy-tetracycline against microorganisms sensitive to tetracycline Infection. The ED ₅₀ (ED = protective dose) for the new compound when infected by Mieroeoccus pyogenes var. Aureus 5 is 3.2 mg / kg when administered orally and 0.34 mg / kg when administered parenterally. The corresponding ED ₅₀ for tetracycline is 6.4 mg / kg for oral administration and 0.78 mg / kg for parenteral administration.

The 6-methylene compounds can be administered in a variety of forms analogous to the parent compounds are processed. They are useful in human therapy, and they are therapeutically in Feedstuffs or as growth stimulants, useful in veterinary medicine and in agriculture.

For the conversion of the Ila-halo-tetracycline hemicetal in 11 a - halogen - 6 - methylene tetracycline the 11 a-chlorotetracycline hemiketals are preferred, since these connections appear to be fairly stable in general.

The following table 111 summarizes the activity of tla-halogen-o-methykn-tetracyclines against a variety of disease-causing microorganisms emschBeßtech antibiotic-resistant strains. The MiadestbemmzentratkMi (MIC) is determined from the known technique of the dilution grater

Table IH

Mkroooccus pyogenes var. Aurens Streptococcus pyogenes Streptococcus faecaEs Diplococcus pneomoniae

MIC

50 (25 p)

1Z5

25th

25th

organism MIC (; / ml) - 6.25 , Erysipelothrix rhusiopathiae 25th Corynebacterium diphtheriae > 100 6.25 Listeria monocytogenes 25th 25th Bacillus sublilis 3.12 12.5 Lactobacillus casei 100 (50 p) 0.78 Bacterium ammoniagenes 12.5 0.39 Aerobacter aerogenes > 100 > 100 Eschcrichia coli > 100 100 Proteus vulgaris > 100 ¹⁵ Pseudomonas aeruginosa > 100 Salmonella gallinarum > 100 100 Salmonella pullorum 100 (ρ) 50 Klebsiella pneumoniae > 100 ²⁰ Neisseria gonorrhoeae 12.5 Hemophilus influenzae 6.25 Shigella sonnei Bruceila bronchiseptica 25 Malleomyces mallei Vibrio comma Pasteurella multocida Streptococcus adalactiae 30 Mycobacterium 607 Mycobacterium berolinense Candida albicans Sarcina Iu tea 35 antibiotic-resistant strains of Mieroeoccus pyogenes var. Aureus 376 *) 400 **)

^. *) Telracyclinrcsistenl at a concentration below 100 / ml. ** "**) Tetracydine-resistant at a concentration below 50 y / ml.
(ρ) = partial inhibition.

The following experiment shows the antibiotic effectiveness of 11a-chloro-6-methylene-tetracycline hydrochloride in comparison to the known 6-desmethyl-6-deoxy-tetracycline hydrochloride. 165 mg of 1 l-chloro-6-methylene-tetracycline hydrochloride were administered orally to each of 10 test subjects and the blood level values were determined at certain time intervals. The same was done with 150 mg of the known 6 - desmethyl - 6 - deoxytetracycline hydrochloride. The doses administered are based on equal amounts of tetracycline hydrochloride The mean blood level values (r / ml)

SS can be seen from the following table IV:

TabeflelV

te (StBBdCO)

Ua-Orior-6-niethylewiracycfinfeydrD-ddorid

6-desmethyl-6-desaxy-

cnlond (known)

24

48

2.M4 1.4 " 1.406 0.408 O

1.293

1.1 Π 0.440
a.052

The advantageous properties of the inventive Compounds that can be produced 6-methylenetetracyclines are thus causal through the properties of the compounds according to the invention. This is also shown in the fact that one z. B. at Hydrogenation of a 1 la-Halogcn-o-deoxy-o-methylenetetracycline with elimination of hydrogen halide 6-deoxytetracycline is obtained, compared to a number of strains are less antibiotic than those from the compounds according to the invention 6-MethyIentetracycline producible (see the table I, experiments 1 and 2). Also the elimination of hydrogen halide from 1 la -halogen -6-deoxy-6-desmethyltetracyclines by catalytic hydrogenation leads to an o-deoxy-o-desmelhyltelracycIin, the shows inferior untibiotic properties to a number of strains than those from the 6-methylenetetracycline compounds according to the invention which can be prepared (cf. Table 1, Experiment 6).

The beneficial properties of 6-methylenetetracyclines are probably due to the 6-position methylene group, which are already present in the Connections is in place.

The following examples illustrate the invention.

example 1

a) 11 a-fluorotetracycline-6,12-hemikelal
(Source material)

30th

45 ml are added to a suspension of 20 g of tetracycline base in 8 (X) ml of water, cooled to 0 ° C. (2 equivalents) 2N sodium hydroxide solution. The tetracycline dissolves into a solution with a pH approximately 11. Perchloryl fluoride is left Bubble through the stirred solution, which is kept under nitrogen, until the pH is the Mixture is approximately 7. Heavy precipitation begins between a pH value of 8 and 8.5 to build. The excess perchloryl fluoride is flushed out with a stream of nitrogen and filtered the almost white, crystalline material, washes it with water and dries it in a vacuum at room temperature, 7.9 to 8.5 g of the product are obtained. The ultraviolet absorption spectrum shows Maxima at 267 and 340 ηΐμ; the ultrared absorption spectrum shows no carbonyl absorption below 6 microns. The biological test against Klebsiella pneumoniae shows an activity of 8 μg / mg, based on tetracycline. The elemental analysis gives the following values:

C ₂₂ H ₂₃ N ₂ O ₈ F

Calculated ..
found ...

H ₂ O:

. C 54.95. H 5.20, N 5.83%:
. C 54.97. H 5.19, N 5.85%.

Example 2

a) I la-fluoro-5-hydroxyietracycline-6,12-hemiketal (Source material)

It is added to a mixture of 6.9 g of anhydrous oxytetracycline base dissolved in 285 ml of methanol and cooled in an ice bath, 1 equivalent in sodium methylate-methanol solution. The yellow sodium salt was cancelled. Perchloryl fluoride is allowed to pass through the

■ ° mixture of pearls. The sodium salt dissolves again. When the mixture approaches the neutrality point a heavy precipitate begins to form. The excess chlorine fluoride is flushed out with a stream of nitrogen, the pro-

'5 duct filtered, washed with cold methanol and dried in vacuo at room temperature, 5.1 g of pale yellow crystals being obtained. The ultrared absorption spectrum shows no carbonyl absorption below 6 microns. The biological trial against Klebsiella pneumoniae gives an activity of 4 μg / mg, based on tetracycline. The ultraviolet absorption spectrum shows maxima at 265 and 336 μm: the elemental analysis shows after recrystallization of the product from water gives the following results:

C ₂₂ H ₂₃ O ₉ N ₂ F-2H ₂ O:

Calculated ... C 51.4, H 5.25, N 5.5%:
found .... C 51.2, H 5.3, N 5.7%.

b) Ila-fluoro-S-hydroxy-ö-dcsoxy-o-desmethyl-6-methyIentetracycline

b) Ila-F] uor-6-deoxy-6-desmethyl-6-methylenetetracycline

250 mg of sodium fluorotetracycline-6,12-hemiketal are stirred into 2 ml of 63% strength aqueous perchloric acid. The solid substance dissolves when heated to 60 to 65 ° C. for 15 minutes. The mixture is then cooled and water is added. 1 la-fluoro-6-deoxy-6-desmethyl-6-methylenetetraeydin is obtained as the perchlorate salt. and ultrared analysis the same absorption as the compound of Example 3 b).

6 g lla- fluorine - 5 - hydroxytetraeycline - 6.12 - hemiketal are dissolved in 45 ml of hydrogen fluoride and left to stand overnight at room temperature. The solvent wiird removed in a stream of nitrogen and the residue dissolved in methanol. The solution becomes Given ether and the precipitate filtered off and dried. 5.1 g of Ila-fluoro-5-hydroxy-

6 - deoxy - 6 - desmethyl - 6 - methylene - tetracycline with ultraviolet absorption maxima in 0.01-n methanolic HCl at 236, 268 and 365 ηΐμ and an ultraviolet absorption maximum in potassium bromide at 5.7 μ.

Example 3

a) 1 la-Chlortetracycline-o ^ -hemiketal
(Source material)

800 mg of N-chlorosuccinimide are added to a solution of XL g of anhydrous tetracycline in 25 ml of the DimethylätheisdesÄthylenglykoIs with stirring in order to dissolve the reagent. You KBt the mixture

Sieve for 7 minutes and then dilute with 25 ml of water. 873 mg of the product crystallize as white needles. The biological examination of the product shows a tetracydin activity of about 4 μg / mg against Klebsiella pneumoniae. The ultrared absorption spectrum shows no carbonyl bands between 5 and 6 p. The ultraviolet absorption spectrum shows maxima at 267 and 340 to 342 mju

The crystalline hydrochloride of this product is obtained by dissolving in excess aqueous Hydrochloric acid (pH approximately 1) and genier drying the mix.

b) 11 a-chloro-o-deoxy-o-desmetbylo-methjfcntetracycline

Dissolve lla-chlortetracycline-6,12-hemJketaI in liquid hydrogen fluoride (in a ratio of 2 g / 15 mL) at 0 ⁰ C. The mixture is maintained for 10 to 15 minutes at this temperature. The hydrogen fluoride is then evaporated off. To obtain the hydrofluoride salt of 1 la-chloro-6-deoxy-6-desmethyl-6-methylenetetracycline as a solid product, the residue is triturated with ether and the solid product is recrystallized from methanol.

On the other hand, 10 g of the crude hydrofluoride product can also be obtained by heating and stirring in Dissolve 350 ml of water. An equal volume of concentrated hydrochloric acid is added to the clear solution, whereupon the product crystallizes as the hydrochloride salt. Elemental analysis of the hydrochloride salt gives the following values:

C ₂₂ H ₂₂ O ₇ N ₂ Cl ₂ :

Calculated:

C 53.11, H 4.56, N 5.63, CP 7.13%;
found:

C 52.62, H 4.63, N 5.54, CP 6.84%. * 5

The ultra-red analysis of the product as the hydrochloride salt (KBr) in 1% concentration gives a carbonyl absorption at 5.70 µ and also the following essential maxima: 6.1; 6.23; 6.36; 6.45 (shoulder); 6.91; 7.85; 8.14; 8.55; 10.22; 10.55 and 10.89.

The biological examination with the product (Klebsiella pneumoniae) shows an oxytetracycline activity of 50 to 100 γ / mg. The ultraviolet analysis of the sample in 0.01 n-methanol-HCl shows maxima at 3.76, 278 and 242 ηΐμ. The product shows an R _r value of 0.02 to 0.3 in the following system:

40

Mohiic phase

Toluene pyridine (saturated
with a pH 4.2 buffer)
(20: 3)

Immobile phase

pH 4.2 buffer (aqueous)

45

When analyzing the paper chart with ultraviolet light, the stain of the product fluoresces not strong; however, it shows strong fluorescence after spraying with aqueous sodium hydrosulfite.

If the product is boiled in a mixture of methanol and concentrated hydrochloric acid, this becomes unchanged product won. Under the same conditions, the starting compound is converted into a Compound converted, which is probably 1 la-chloroisotetracycline, and which is used in the treatment with aqueous sodium hydrosulfite gives isotetracycline.

One can also use IIIa-chloro-o-deoxy-o-desmethyl-6-methylentetracycline hydiofluoride Dissolve in water and adjust the solution to pH 5. The precipitating amphoteric compound is filtered and dried. The amphoteric compound is dissolved in methanol, which is 1 molar equivalent of hydrogen chloride contains, and the hydrochloride salt is precipitated by adding ether. By replacing the hydrogen chloride other mineral acids can be used to obtain the corresponding other salts.

For example, hydrobromide, sulfate, hydroiodide, Nitrate and phosphate are produced this way.

You can also use organic carboxylic acids, such as tartaric acid, citric acid, malic acid, benzoic acid, Use glycolic acid, gluconic acid, gulonic acid, succinic acid, or acetic acid. The one with pharmaceutical Salts formed from perfect acids can be used for therapy; those with pharmaceutical Incorrect acids formed salts are for cleaning the products and for the production of pharmaceutically acceptable salts.

If, under the same conditions, 85% phosphoric acid is used instead of sulfuric acid, the phosphate salt of Ila-chloro-6-deoxy-o-desmethyl-ö-methylenetetracycline is obtained in this way.

Example 4

a) 7.1 la-D ^; chlortetracycline-6,12-hemiketal
(Source material)

A mixture of 2.4 f. Anhydrous 7-chlortetracycline, 800 mg of N-chlorosuccinimide and 25 ml of 1,2-dimethoxyethane 2 '/ ₂ minutes, after which 100 ml of ether are added and then 300 ml of hexane. The precipitate that forms is collected by filtration, washed with hexane and dried.

2.1 g of 7.1 la-dichlorotetracycline-o. ^ - hemiketal are obtained, which shows no activity against Klebsiella pneumoniae.

b) The dehydration of the hemiketal with the formation of 7,1a-dichloro-6-deoxy-6-desmethyl-6-methylene tetrachloride is carried out in the same way as in Example 3 b).

Example 5

a) 1 la-chloro-S-hydroxytetracycline-o. ^ - hemiketal
(Source material)

α) 10g 5 - Hydroxytetracycline - hydrochloric! are dissolved in 200 ml of 50% aqueous 1,2-dimethoxyethane, which has previously been cooled to 5 ° C., with the aid of 3.5 ml of triarhanolamine. Immediately thereafter, 4.4 g of powdered N-chlorosuccinimide are added. The temperature of the reaction mixture, cooled by an ice / water bath, increases to 12 ^° C. during the addition. The mixture, in which the product crystallizes out very quickly, is stirred for 10 minutes on an ice bath, filtered and washed with a sufficient amount of acetone to darken To remove impurities. The yield of 1 la-chloro-S-hydroxytetracycline-o ^ -hemiketal is 5.0 g (50%).

An analytically pure product is obtained by recrystallization from methanol / hydrochloric acid. The product shows ultraviolet absorption maxima in 0.01-n ethanolic HCl at 26: 5; 338 ηΐμ; log / 4.39 and 3.67.

C ₂₂ H ₂₃ O ₉ N ₂ Cl CH ₃ OH HCl:

Calculated:

C 49.0, H 5.0, N 5.0, Cl "6.3, Cl 12.6,

CH ₃ O 5.5%;
Findings:

C 49.0, H 5.0, N 4.8, Cl "6.3, Cl 12.7,

CH ₁ O 5.4%.

609 638/84

β) 10 g of S-hydroxytetracycline hydrochloride are dissolved in UOmI 70% aqueous acetone. The solution is cooled to 4 ° C., and 6 g of sodium p-toluenesulfonchloramide ("Chloramine T") in 40 ml of 70% strength AcetGn are added. The mixture is stirred cold for 20 minutes, filtered and the filter cake washed four times with 10 ml portions of 70% aqueous acetone to give 6.66 g of Ua-chloro-5-hydroxytetracycline-6.12-hemiketal, the same Ultraviolet absorption spectrum shows like the product according to Example 5 a) α).

γ) 20 g of S-hydroxytetracycline hydrochloride are dissolved in 148 ml of aqueous acetone 70%, cooled to -20 ⁰ C and then treated with 7.0 ml of triethylamine and then with 6.1 g of p-Toluolsulfoudiehloramid ( "dichloramine T") in 20 ml of 70% aqueous acetone, which was ^{precooled to -20 0} C, treated. After 5 minutes, 225 ml of water at 0 ° C are slowly added, and after 14 minutes the resulting paste is filtered and the filter cake is filled with two 20 -ml- portions of 70% aqueous acetone and three 20-ml portions of acetone. The residue is ^{dried at 35 °} C. in vacuo, giving 10.20 g of 1 l-chlorine-5-hydroxyletracycline-6,12-hemiketal, which shows the ultraviolet absorption maxima in 0.03-n methanolic HCl at 266 and 339 ηΐμ.

ö) 50 g S-hydroxytetracycline hydrochloride are dissolved in 370 ml of 70% aqueous acetone, and at - 25 C chilled. 17.5 ml of triethanolamine are added, followed by 9.6 g of trichlorocyanurtriamide ("trichloromelamine"), which is dissolved in 50 ml of 70% aqueous acetone precooled to 20 ° C. After 5 minutes, 562 ml of water are slowly added at OC , and after 14 minutes the slurry obtained is filtered at -8 C. The filter cake is washed with two 50 ml portions of 70% aqueous acetone and three 50 ml portions of dry acetone are dried in vacuo at 35 C, whereby 25 , 35 g of lla-chloro-5-hydroxytetracycline-6,12-hemiketal is obtained, which shows the ultraviolet absorption maxima in 0.03-n methanolic hydrochloric acid at 266 (E | l = 346) and 340 ηΐμ (E | * = 81).

If you work in the same procedure, but replace "trichloromelamine" with other halogenating agents, this gives the following results:

The product is obtained by vacuum filtration in an ice bath, washed with acetone and air-dried. The yield of crystalline Ila-chloro-o-deoxy-o-des-methvl-6-methylene-5-hydroxytetracycline hydnodide, which contains 2 mol of crystal acetone, is 3.4 g (47%). The product shows ultra-red absorption maxima at 570 (in KBr) and 5.86 μ (in acetone) and ultraviolet absorption maxima in OPl-ii methanohic HCl at 223, 271 and 374 πΐμ; log ε 4.47, 4.27 and 3.56.

C ₂₂ H ₂₁ N ₂ O ₈ Cl Hl 2CH ₃ COCH ₃ :

Calculated:

C 46.6, H 4.8, N 3.9, Cl 4.9, I neutralization equivalent 721;

found:

C 46.4, H 4.8, N 3.8, Cl 5.0 ^ I. Neutralization equivalent 706.

The product can also be obtained from liquid hydrofluoric acid as the p-toluenesulfonic acid salt Instead of the acetone and hydrogen iodide mixture, previously described, 50 ml of isopropyl alcohol, 50 ml of acetone and 5 g of p-toluenesulfonic acid are then used. After washing with acetone, 5.6 g (81%) of the air-dry material are then obtained

17.6%;

17.5%;

Halogenation agent

13.2 g of 1,3-dichloro-5,5-dimethylhydride

9.8 g of trichloroisocyanuric acid

Yield of UV absorption hemicctal maxima

(g) (mn)

22.35
22.75

265 and 340 268 and 360

b) 1 la-chloro-o-deoxy-o-desmethyl-o-methylene-5-hydroxytetnicycline

50

55

60

a) 5 g of 1 Ia-chloro-5-hydroxytetracycline-6,12-hemiketal are dissolved in 15 ml of liquid hydrogen fluoride. The reaction mixture is stirred at 0 to 5 ° C. for 3.5 hours and then poured into 100 ml of cold acetone which contains 7.5 mm of 7% strength aqueous hydriodic acid and a few seed crystals of the product. It is stirred for 45 minutes for a rf) 1Π ml (205 g) of concentrated sulfuric acid is cooled to 15 ⁰ C and diluted under stirring with 14 ml of water. After Widerabkühlen at 15 C solution of acid over 30 minutes in portions 51g are lla-chloro-S-hydroxytetracycline-6-hemiketal given 12 to 90%, while maintaining the temperature by cooling to 18 to 22 ^C C. The last clear, dark red solution is stirred for 1 hour at 20 to 25 C and then a mixture of 1070 ml of acetone and 1.00 g of ascorbic acid (antioxidant) and 75 ml of 47% hydroiodic acid and seed crystals of lla-chloro-S- hydroxy-o-deoxyo-desmethyl-o-methylene-tetracycline added. The addition lasts 20 minutes, during which the temperature is kept at 5 to 10 ° C. A little acetone is added and the rapidly cooled solution is stirred for 30 minutes until a crystal paste forms. The product is collected by filtration, washed with acetone. and dried under reduced pressure at 40 to 45 C, 37 g of Ila-chlorine

5 - hydroxy - 6 - deoxy - 6 - desmethyl - 6 - methylene tetracvcline obtained, which shows the same ultra-red absorption maxima as that obtained in Example 5b) α) Product obtained using hydrogen fluoride.

Example 6

7,11 a-Dichloro-S-hydroxy-o-deoxy-ö-desmethyl-6-methylene-tetracycline

Method a

Add to 5 g of lla-chloro-o-deoxy-o-desmethyl-

6 - methylene - 5 - hydroxytetracycline - hydroiodide in 15 ml of liquid hydrogen fluoride, cooled to ice bath temperatures, 1.5 g of N-chlorosuccimmide. The solution is stirred at ice bath temperature for IV2 hours. The crude product is precipitated by adding 500 ml of ether and recovered by filtration.

The crude product is taken up in methanol at room temperature and the insoluble material is filtered off the filtrate is treated with activated charcoal and filtered

and concentrated under reduced pressure. The residue obtained is dissolved in dilute hydrochloric acid added, from which the product crystallizes as the hydrochloride. The ultraviolet absorption analysis in 0.01 n-HCl in methanol gives the following values:

X ^ _x 239 mji, El * 352; J _-1 378 πΐμ, E | * 60;
Bend 2.58 πΐμ. EJ * 324.

The ultrared absorption analysis shows the major bands at 5.7, 6.0 and 6.9 microns.

IO

Method B.

5 g of IIIa-chloro-5-hydroxytctracycline-6,12-hemiketal are added to 15 ml of liquid hydrogen fluoride at ice barite temperature. After Z ¹ I ₂ hours of stirring at ice-bath are added the same amount by weight of N-chlorosuccinimide and the procedure following the procedure of Method A for the recovery of the product.

Another and somewhat simpler method of up-* ° processing is as follows: After removing the greatest amount of liquid hydrogen fluoride was added 100 ml of water and then 5 g / S-naphthalenesulfonic to. The product precipitates out as the salt of naphlhalinsulfonic acid and is collected by filtration.

Another work-up method involves diluting the original reaction mixture with 6 to 7 parts by volume of water and subsequent dropwise addition of concentrated acid to the Precipitation of the perchlorate and 0-naphthalene sulfonate salts, as described above. The crude perchlorate salt thus obtained crystallizes as long needles from isopropanol, which in the ultraviolet analysis maxima at 260 and 377 ηΐμ and a bend (inflection) at 260 ηΐμ show. The ultrared absorption analysis gives Peaks at 5.7, 6.0, 6.26, 6.55, 6.88, 7.2, 7.85 and 8.35 microns.

B e i s ρ i e 1 7

a) 11 a-Bromo-S-hydroxytetracycline-o, 12-hemiketal
(Source material)

250 ml of 5-hydroxytetracycline base are dissolved in 10.2 ml of a benzene: 1,2-dimethoxyethane mixture Dissolved 1: 1 to 35 ° C. This is done drop by drop within a solution of 0.51 ml of bromine for 1N minutes given in 10 ml of a benzene: 1,2-dimethoxyethane mixture 1: 1. A discolored one will soon divorce Precipitation from. The mixture is stirred for 30 minutes at 35 ° C., the reaction mixture is filtered and washed the filter cake with benzene / dimethoxyethane. The somewhat hygroscopic solid is dissolved in 10 ml of benzene / Dime1: slurried hoxyethane, obtained by filtration and dried in vacuo, whereby one 173 mg of crystalline Ila-bromo-5-hydroxytetracycline-6,12-hemiketal receives. Another 52 g are obtained by leaving the combined filtrates at overnight Let stand room temperature, filter off, wash the filter cake with ether and then in vacuo dries The product shows no ultrared absorption in the range of 5 to 6 μ (measured in potassium bromide) and iced ultraviolet absorption maxima in 0.01-N methanolic HCl at 272 and 420 πΐμ. (EJi = 184). If the product is boiled with acetone, it shows an activity against Klebsiella pneumoniae of 600 μg / mg, which indicates the conversion to 5-hydroxytetracycline.

b) 1 la-bromo-S-hydroxy-o-deoxy-o-desmethyl-6-methylene-tetracycline

The dehydration of the hemiketal to form the tetrafluoride of Ila-bromo-5-hydroxy-6 - Desoxy - 6 - desmethyl - 6 - methylentetracycline is made by adding 150 mg of the hemiketal Dissolve the hydrogen fluoride in 1 ml of hydrogen fluoride and let stand at 0 ° C. for 1 hour removed and the residue was washed with ether. The product shows an ultra-red absorption maximum in Potassium bromide at 5.7μ.

The hydrochloride salts of the above-described lla-halo-tetracycline-o. ^ - hemiketale are prepared by dissolving the free base in water, which contains an equimolecular amount of hydrogen chloride, and (the resulting solution is freeze-dried. Similarly, salts with sulfuric acid, hydrobromic acid and phosphoric acid.

Claims (1)

Claim: Compounds Αχ general formula CH ₂ EN (CH ₃ ), -OH CONH ₂