AT226366B - Process for the production of new penicillin derivatives - Google Patents

Description

  

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  Process for the production of new penicillin derivatives
The invention relates to new types of penicillins which are derivatives of 6-aminopenicillanic acid: this has the following structural formula:
 EMI1.1
 
It has now been found in accordance with the invention that by using an acid chloride, bromide, anhydride or a mixed anhydride of a trisubstituted acetic acid, a particularly useful class of derivatives thereof can be obtained.



   These compounds are useful as antibacterial agents, as nutritional additives for animal feed, as agents for treating mastitis in cattle, and as therapeutic agents in poultry and other animals, as well as humans, in the treatment of infectious diseases particularly caused by gram-positive bacteria.



   Antibacterial agents such as benzylpenicillin have been shown to be highly effective in the therapy of infections from gram positive bacteria; However, these agents have the serious disadvantage that they are ineffective against many so-called resistant bacterial strains, for example penicillin-resistant strains of Staphylococcus aureus (Micrococcus pyogenes, var. aureus), which produce penicillinase. Many of the compounds according to the invention, in addition to their strong antibacterial activity, show resistance to destruction by penicillinase or are active against benzylpenicillin-resistant bacterial strains.



   The new penicillin derivatives of the invention have the following general formula:
 EMI1.2
 
 EMI1.3
 and each represent a halogen atom or an optionally substituted alkyl, alkenyl, aryl, aralkyl, cycloalkyl or heterocyclic group, or R and R together with the carbon atom,

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 to which they are bound, form an aromatic, cycloaliphatic or heterocyclic ring system and R is one of the groups listed above under Rl and R or the group OR4. or SR4. denotes, where R * denotes an alkyl, alkenyl, aryl, aralkyl or cycloalkyl group, as well as their therapeutically useful salts.



   Suitable salts are, for example, compatible metal salts such as sodium, potassium, calcium and aluminum, the ammonium salt and substituted ammonium salts, for example salts of such compatible amines, such as trialkylamines, including triethylamine, procaine, dibenzylamine, N-benzyl-ss-phenethylamine, 1 -Ephenamine, N, N'-dibenzylethylenediamine, dehydroabietylamine, N, N'- bis-dehydroabietylethylenediamine and other amines which were used to form salts with benzylpenicillin.



   A preferred class of novel penicillin derivatives according to the invention has the formula (II) given above, in which R is a substituted aromatic ring, at least one of the substituents being a hydroxyl group and R and R being identical or different and each being the same group as R or optionally one are substituted alkyl, aryl or aralkyl groups.



   Another class has the structural formula (II), where R is a substituted or unsubstituted benzene ring, R2 is an alkyl group or an alkoxy group, each containing 1-6 carbon atoms and R? means one of the groups defined under R and R2.



   Another group has the general formula:
 EMI2.1
 wherein X, Y and Z are identical or different and each have a hydrogen, nitrogen or halogen atom or an amino, alkylamino, dialkylamino, acylamino, alkanoylamino, alkyl, alkoxy, hydroxy, sulfamyl , Benzyl, cyclohexyl, cyclopentyl or trifluoromethyl group and where furthermore A and B are identical or different and each denotes an alkyl, phenyl or phenylalkyl group.



   According to the invention, a process for the production of new types of penicillin of the general formula (II) is provided, wherein 6-aminopenicillanic acid with an acid chloride, bromide, anhydride or with a mixed anhydride with another acid, from a trisubstituted acetic acid of the general formula:
 EMI2.2
 
 EMI2.3
 PH value of about 6 to 9 is implemented.



   Examples of carboxylic acids of the general formula (IV) are given below; the abbreviations "Ph", "Me", "Ät" are used for phenyl, methyl and ethyl groups:

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 EMI3.1
 
 EMI3.2
 
 EMI3.3
 
 EMI3.4
 Me H H Me Me Me Me CHMe2 CHMe2 Me
 EMI3.5
 

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 b) Diarylalkyl type
 EMI4.1
 c) Aryldialkyl type
 EMI4.2
 d) trialkyl type
 EMI4.3
 
 EMI4.4
 

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 EMI5.1
 f) Diaryl (thio-substituted) type
 EMI5.2
 g) Dialkyl (oxy-substituted) type
 EMI5.3
 h) Dialkyl (thio-substituted) type
 EMI5.4
 

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 EMI6.1
 i) Arylalkyl (oxy- or thio-substituted) type
 EMI6.2
 j) diarylheterocyclo-type
 EMI6.3
 k) 1-Arylcycloalkyl-Type
 EMI6.4
   l)

     9-substituted 9-fluorenyl-type
 EMI6.5
 

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 EMI7.1
 
 EMI7.2
 R = H or Me m) Miscellaneous
 EMI7.3
 
In cases where the acid contains a group, for example an amino or alkylamino group, which can react with an acylating agent, such group can be protected from the formation of the acid chloride, bromide, anhydride or mixed anhydride in the usual manner . The subsequent removal of the protective group to form the free amino-substituted or alkylamino-substituted penicillin can be effected by catalytic hydrogenation, for example with palladium or platinum on barium carbonate or carbon.

   Suitable protecting groups are the trityl group and groups of the general formula R "-O-CO-, where R" denotes an allyl, benzyl, substituted benzyl, phenol or substituted phenyl group. On the other hand, those penicillin derivatives in which the substituent is an amino group can also be prepared by first preparing the corresponding nitropenicillin and then hydrogenating it in the usual way to give the amino derivative.



   If an acid chloride, bromide or anhydride is used in the process according to the invention, this is prepared from the corresponding trisubstituted acetic acid by the process described in the literature.



   A process for the preparation of a compound provided by the invention with the aid of a mixed anhydride with ethoxy- or isobutoxycarboxylic acid consists in that an acid of the general formula (IV) with isobutyl chloroformate and a tert. Hydrocarbon or aliphatic amine such as triethylamine in an anhydrous, inert and preferably water-miscible solvent such as. B. p-dioxane and, if desired, pure dry acetone is mixed for about 30 minutes in the cold. A cooled solution is then added to this solution of the mixed anhydride.

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 Solution of 6-aminopenicillanic acid and tertiary amine, for example triethylamine, in a solvent such as water, for example.

   The reaction mixture is stirred for about 1 hour during which time the substituted ammonium salt of the desired product forms. The mixture can then, if desired, be extracted at alkaline pH (approximately at a pH of 8; for example, an aqueous bicarbonate solution can be used to adjust the pH if necessary), and the like. zw. With a water-immiscible solvent, such as. B. ether to remove unreacted starting material. The product in the aqueous phase is then converted into the free acid, preferably in the cold under an ethereal layer by adding dilute mineral acid, for example
Sulfuric acid, up to a pH of 2.

   The free acid is then extracted by a water-immiscible neutral organic solvent such as ether and the extract is, if desired, quickly washed in the cold with water and then dried, for example, with anhydrous sodium sulfate. The product contained in the essential extract in the form of the free acid is then through
Treatment with the appropriate base, for example a free amine such as procaine base or a
Solution of potassium 2-ethylhexanoate in dry n-butanol, in any desired amine salt or
Metal salt converted. These salts are usually insoluble in solvents such as ether and can be obtained in pure form by simple filtration.



   Another method of making an essential solution is the acidic form of a compound of the
The invention consists in first preparing an aqueous solution of 6-aminopenicillanic acid and sodium bicarbonate, to which an acid chloride of an acid of the general formula (IV) is added and shaken vigorously at room temperature, for example 20-60 min. The mixture is then extracted with ether to remove unreacted or hydrolyzed starting materials. The solution is then acidified (preferably in the cold) to a pH value of 2, for example with dilute sulfuric acid, and the corresponding free acid is extracted in ether.

   This essential extract is dried, for example with anhydrous sodium sulfate, the desiccant is then removed and a dry ethereal solution is obtained from which the product can easily be isolated, preferably in the form of an ether-insoluble salt such as the potassium salt. This method is used when the acid chloride reacts faster with a primary amine than with water, which can be determined by a simple experiment. In this process, the corresponding acid bromide or acid anhydride can be used instead of the acid chloride.



   Since some of the antibiotic substances obtainable by the process according to the invention are relatively unstable compounds which are easily subject to chemical changes and thus lose their antibiotic effectiveness, reaction conditions should be selected which are sufficiently mild to avoid the decomposition of the substances. These reaction conditions will, of course, depend in large part on the reactivity of the reagents used. In most cases a compromise has to be made between using very mild conditions for a long period of time and using more severe conditions for a shorter period of time with the possibility of some of the antibiotic substances degrading.



   The temperature during the preparation of the penicillanic acid derivatives should generally not exceed 30 ° C. and in many cases room temperature is suitable for this. Since the use of strongly acidic or alkaline conditions is to be avoided in the method according to the invention, the method can advantageously be carried out at a pH value of 6 to 9 and this pH range can advantageously be achieved by using a buffer, for example a solution of sodium bicarbonate or a sodium phosphate buffer.

   In addition to an aqueous reaction medium including filtered fermentation broths or aqueous solutions of crude 6-aminopenicillanic acid, organic solvents which do not contain any reactive hydrogen atoms can also be used. Examples of such inert solvents are dimethylformamide, dimethylacetamide, chloroform, acetone, methyl isobutyl ketone and dioxane. It is often very advantageous to add an aqueous solution of a salt of 6-aminopenicillanic acid to a solution of the acylating agent in an inert solvent, preferably in an inert solvent which is miscible with water, such as acetone or. Dimethylformamide.

   Vigorous stirring is of course advisable if more than one phase is present, for example a solid and a liquid or two liquid phases.



   At the end of the reaction, the products are isolated, if desired, according to the methods known for benzylpenicillin and phenoxymethylpenicillin. The product can be extracted with diethyl ether or n-butanol at acidic pH and then obtained by lyophilization or by conversion into a solvent insoluble salt, such as by neutralization with an n-butanol solution of potassium - 2-ethylhexanoate or the product can be obtained from aqueous Solution as a water-insoluble salt of an amine

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 EMI9.1
 
 EMI9.2
 
<tb>
<tb> Compound <SEP> Minimum <SEP> inhibitory concentration
<tb> more resistant <SEP> more resistant <SEP> more resistant
<tb> strain <SEP> 1 <SEP> strain <SEP> 2 <SEP> strain <SEP> 3
<tb>
<tb> Ph. <SEP> CH. <SEP> CO. <SEP> NH. <SEP> CH-CH <SEP> C. <SEP> Me <SEP> 1 <SEP>: <SEP> 4000 <SEP> 1 <SEP>:

   <SEP> 400000 <SEP> 1 <SEP>: <SEP> 1000000 <SEP>
<tb> I <SEP> I <SEP> I <SEP> (250 <SEP> Jlg / ml) <SEP> (2, <SEP> 5pg / ml) <SEP> (1 <SEP> pg / ml) < SEP>
<tb> CO-N ----- CH. <SEP> CO <SEP> H <SEP>
<tb> penicillin <SEP> G
<tb> \
<tb> PhC. <SEP> CO. <SEP> NH. <SEP> CH-CH <SEP> C. <SEP> Me <SEP> 1 <SEP>: <SEP> 3200000 <SEP> 1 <SEP>: <SEP> 6400000 <SEP> 1 <SEP>: <SEP> 6400000 <SEP>
<tb> 1 <SEP> 1 <SEP> 1 <SEP> (0, <SEP> 3 <SEP> p <SEP> glm <SEP> 1) <SEP> (0, <SEP> 16 <SEP> Mg / ml) <SEP> (0, <SEP> 16 <SEP> pglml) <SEP>
<tb> Co-N-Ch. <SEP> Co2H
<tb>
 

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The following examples are intended to explain the invention without restricting it thereto. (These examples are grouped according to the previously given classification system for trisubstituted acetic acids.) A) Triaryl type.



     Example 1: Triphenylmethylpenicillin.



   (i) Crude sodium salt.



   A solution of 0.8 g of triphenylacetyl chloride in 10 ml of chloroform became one over 5 minutes
Suspension of 0.5 g of 6-aminopenicillanic acid (72% pure) in 20 ml of chloroform and 1 ml of triethylamine are added. After stirring for one hour at room temperature, the solution was filtered and the solvent was evaporated in vacuo at low temperature. The semi-solid residue was stirred into 50 ml of n-butanol, 30 ml of water were added and the pH of the aqueous phase was brought to 2 with n-hydrochloric acid. It was then shaken, the aqueous layer was discarded and the butanol phase was washed two more times with very dilute hydrochloric acid.

   The penicillin was obtained from the butanol layer by shaking it with enough sodium bicarbonate solution to obtain a neutral aqueous phase (PH 7) and separating the latter. The water was evaporated off at low temperature and reduced pressure, leaving the crude sodium salt of triphenylmethylpenicillan as a pale yellow solid (0.9 g). The purity was 55 yes, determined by a colorimetric comparison with hydroxylamine against a benzylpenicillin standard. The product inhibited Staphylococcus aureus at a concentration of 0.3 g / ml, the benzylpenicillin-resistant staphylococcal strain 1 at 0.6 g / ml and the benzylpenicillin-resistant staphylococcal strain 2 at 0.6 ug / ml.



   (ii) benzylamine salt.



   150 mg of the crude sodium salt of triphenylmethylpenicillin, prepared according to (i), were suspended in 15 ml of water, covered with an equal volume of ether and acidified to pH 2 with dilute hydrochloric acid. This was followed by shaking, the free penicillic acid passing into the ethereal layer. The ether solution was separated off, washed with water, dried over anhydrous sodium sulfate, filtered and treated with 2 ml of a 2% solution of benzylamine in ether. The benzylamine salt of penicillin precipitated as an oil, which later slowly solidified, probably taking up crystal water from the atmosphere. The resulting white powder was collected, washed with ether and dried in a vacuum desiccator. Yield 65 mg, m.p. 98-1000 ° C. (decomposition).
 EMI10.1
 
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<tb>



  Calculates <SEP> for <SEP> CgOS, <SEP> 2 <SEP> Ego: <SEP> C <SEP> 66, <SEP> 70/0 <SEP>; <SEP> H <SEP> 6, <SEP> 2%; <SEP> N <SEP> 6, <SEP> 70 / o, <SEP> S <SEP> 5, <SEP> 1% <SEP>
<tb> Found <SEP>: <SEP> C <SEP> 66, <SEP> 9% <SEP> H <SEP> 6, <SEP> 2%; <SEP> N <SEP> 6, <SEP> 8%; <SEP> S <SEP> 4, <SEP> 5%
<tb> (iii) <SEP> cyclohexylamine salt.
<tb>
 



   A solution of triphenylmethylpenicillin in ether was prepared from 180 mg of the crude sodium salt as in (ii). Treatment with 2 ml of a solution of cyclohexylamine in ether gave the cyclohexylamine salt of penicillin as a weakly deliquescent white powder with an indefinite melting point. This was collected, washed with ether and dried in a vacuum desiccator. The yield was 90 mg.
 EMI10.2
 
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  Calculates <SEP> for <SEP> CNOS, <SEP> 2 <SEP> H2O: <SEP> C <SEP> 65, <SEP> ze <SEP> H <SEP> 7.0%, <SEP> N <SEP > 6.8%; <SEP> S <SEP> 5.1%
<tb> Found <SEP>: <SEP> C <SEP> 66.6%; <SEP> H <SEP> 7, <SEP> 40/0 <SEP>; <SEP> N <SEP> 6, <SEP> 9% <SEP> S <SEP> 4, <SEP> 6%
<tb> (iv) <SEP> N-methylmorpholine salt.
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   As described in (ii), a solution of triphenylmethylpenicillin in ether was prepared from 170 g of the crude sodium salt. By treatment with 2 ml of a 21% solution of N-methylmorpholine in ether, the N-methylmorpholine salt of penicillin was precipitated as a colorless oil, which slowly solidified on scratching. The white powder obtained was collected, washed with ether and dried in a vacuum desiccator. The yield was 60 mg, mp = 118-121 ° C. (decomposition).
 EMI10.3
 
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<tb>



  Calculates <SEP> for <SEP> C <SEP> H <SEP> OgN <SEP> S, <SEP> 0, <SEP> 5 <SEP> H2O: <SEP> C <SEP> 66.4%; <SEP> H <SEP> 6.4%; <SEP> N <SEP> 7, <SEP> 0% <SEP> S <SEP> 5.4%
<tb> Found <SEP>: <SEP> C <SEP> 66, <SEP> 60/0 <SEP>; <SEP> H <SEP> 6, <SEP> 30/0 <SEP>; <SEP> N <SEP> 6, <SEP> 9%; <SEP> S <SEP> 5.3%
<tb> (v) <SEP> Pure <SEP> sodium salt.
<tb>
 



   A solution of 18.4 g of pure triphenylacetyl chloride in 360 ml of dry acetone was added over 15 min to a stirred solution of 13 g of 6-aminopenicillanic acid (about 72% pure) in a mixture of 500 ml of a 3% aqueous sodium bicarbonate solution and 150 ml of acetone . The mixture was stirred for 3 hours at room temperature, then washed twice with 600 ml of ether. The aqueous layer was filtered to remove some of the suspended solid and then cooled to -60C, whereupon the product crystallized in glittering plates.

   It was separated off in a cooled centrifuge and then dissolved in 300 ml of tens of aqueous acetone at room temperature and thereafter

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 Cooling to -6 C recrystallized. The pure crystalline sodium salt was separated off as previously described and dried in a vacuum desiccator. 17.1 g of a white powder with a melting point of 100 ° to 1200 ° C. were obtained in this way, the majority of which was obviously present as a monohydrate. Further drying over phosphorus pentoxide at 80 ° C. and 0.5 mm gave the anhydrous salt; Mp 183 to 1900C (decomposition).

   This is hygroscopic and in air the melting point quickly drops to 100 to 1200C.
 EMI11.1
 
 EMI11.2
 
<tb>
<tb> Calculates <SEP> for <SEP> C28H25O4N2SNa: <SEP> C <SEP> 66, <SEP> ilo <SEP> H <SEP> 5, <SEP> 0% <SEP> N <SEP> 5, < SEP> 5% <SEP> S <SEP> 6, <SEP> 4%; <SEP> Na <SEP> 4, <SEP> 5%
<tb> Found <SEP>: <SEP> C <SEP> 65, <SEP> 9%; <SEP> H <SEP> 5, <SEP> 2%; <SEP> N <SEP> 5.2%; <SEP> S <SEP> 6, <SEP> 2lo <SEP> Na <SEP> 4, <SEP> 4% <SEP>
<tb>
 
 EMI11.3
 



   This was produced exactly as described under (v), only the equivalent amount of potassium bicarbonate was used instead of sodium bicarbonate. After isolating and drying as with the
 EMI11.4
 
 EMI11.5
 
<tb>
<tb> Calculates <SEP> for <SEP> CHOSK <SEP>: <SEP> C <SEP> 64, <SEP> 1%; <SEP> H <SEP> 4, <SEP> 8lo <SEP> N <SEP> 5, <SEP> 3% <SEP> S <SEP> 6, <SEP> 1%; <SEP> K <SEP> 7.4%
<tb> Found <SEP>: <SEP> C <SEP> 63, <SEP> 7lo <SEP> H <SEP> 4, <SEP> 8lo <SEP> N <SEP> 5, <SEP> Ilo <SEP> ; <SEP> S <SEP> 5, <SEP> 8%; <SEP> K <SEP> 7, <SEP> 4%
<tb>
 
 EMI11.6
 added for 7 min to a stirred solution of 4.84 g of 6-aminopenicillanic acid in a mixture of 150 ml of aqueous 8.8% sodium bicarbonate solution and 75 ml of acetone.

   The mixture was stirred at room temperature for 3 hours, diluted with 50 ml of water and extracted twice with 100 ml of ether. The aqueous solution thus obtained was covered with 80 ml of ether, acidified to a pH of 2 to 3 with 92 ml of n-hydrochloric acid, shaken and the ether layer separated. The aqueous layer was extracted with two further portions of 40 ml each of ether and the combined ether extracts (which contained the free penicillic acid) were washed with 20 ml of water and then neutralized with 13.5 ml of aqueous 8.4% sodium bicarbonate solution with vigorous shaking. The aqueous layer was separated and evaporated to dryness in vacuo at room temperature. The sodium salt of mono-p-hydroxytriphenylmethylpenicillin was thus obtained as a yellow solid (6.8 g).

   Colorimetric comparison of hydroxylamine versus a benzylpenicillin standard shows that it was approximately 65% pure.



   The product inhibited Staphylococcus Oxford in its concentration of 0.5 J. lg / ml, the Benzylpe-
 EMI11.7
   -p-hydroxytriphenylacetic acid with thionyl chloride in benzene solution with the addition of catalytic amounts of pyridine produced as a viscous oil. The reaction was effected by heating at 60-70 ° C. for one hour and the acid chloride was isolated by concentrating the reaction mixture. Its purity was ensured by model reactions with aniline and ammonia in the presence of water to form anilide and amide, respectively.



    Example 3: Mono-p-methoxytriphenylmethylpenicillin.



   A solution of 673 mg of mono-p-methoxytriphenylacetyl chloride in 5 ml of dry chloroform was added dropwise to a stirred mixture of 432 mg of 6-aminopenicillanic acid, 10 ml of chloroform and 0.84 ml of triethylamine. The mixture was stirred for 21/2 hours at room temperature and any residues which had not yet dissolved were then filtered off. The filtrate was
 EMI11.8
 aqueous phase had a pH of 2.



   The acidic layer was then separated and discarded and the chloroform solution was washed with 20 ml of water. Sufficient 3-day aqueous sodium bicarbonate solution was then added to the chloroform solution so that a neutral emulsion (PH 7) was obtained after vigorous shaking. This was concentrated at low temperature and reduced pressure and the hygroscopic solid residue that remained was dried in a vacuum desiccator. 817 mg of the sodium salt of mono-
 EMI11.9
 

 <Desc / Clms Page number 12>

 



   The mono-p-methoxytriphenylacetyl chloride was prepared from the corresponding acid by heating with thionyl chloride and a trace of pyridine in benzene, after which the solvent and the excess of the reagent was removed in vacuo and the residue was recrystallized from petroleum ether. M.p. 101-1030C.



    At pie 1 4: mono- p-methylthiotriphenylmethylpenicillin.



   A solution of 6.34 g of mono-p-methylthiotriphenylacetyl chloride in 108 ml of dry acetone was slowly added to a stirred solution of 3.89 g of 6-aminopenicillanic acid in 152 μg of 3% aqueous sodium bicarbonate solution and 45 ml of acetone. The mixture was stirred for 4 hours at room temperature, then washed with 2 portions of 200 ml of ether each time. The aqueous phase was covered with 50 ml of ether and brought to a pH of 2 by adding n-hydrochloric acid. After separating the layers, the aqueous phase was extracted with two further portions of 50 ml each of ether. The combined ether solutions containing the free penicillic acid were washed with water
 EMI12.1
 net. Yield 7.44 g, purity about 64%.



   The product inhibited Staphylococcus Oxford at a concentration of 0.025 / μg / ml, the staphylococcal strain 1 with 0.25 μg / ml and the staphylococcal strain 2 with 1.25 μg / ml.



   The mono-p-methylthiotriphenylacetyl chloride was prepared from the corresponding acid by heating with thionyl chloride and a trace of pyridine in benzene, removing the solvent and excess reagent in vacuo and crystallizing the residue from petroleum ether. Mp 103, 5-105 C.



    Example 5: Mono-p-methylsulfonyltriphenylmethylpenicillin.



     4.32 g of 6-aminopenicillanic acid were acylated with 7.68 g of mono-p-methylsulfonyltriphenylacetyl chloride, as described in Example 4, to give the sodium salt of mono-p-methylsulfonyltriphenylmethylpenicillin, an almost white solid. Yield 9.23g, purity about 51%.



   The product inhibited Staphylococcus Oxford at a concentration of 0.6 g / ml, the staphylococcal strain 1 with 2.5 μg / ml and the staphylococcal strain 2 with 2.5 g / ml.



   The mono-p-methylsulfonyltriphenylacetyl chloride was prepared from the corresponding acid by heating with thionyl chloride and a trace of pyridine in benzene, removing the solvent and excess reagent in vacuo and crystallizing the residue from petroleum ether. M.p. 146-1480C.
 EMI12.2
 acylated a white solid (9.7 g).



   The product inhibited Staphylococcus Oxford at a concentration of 12.5 μg / ml, the staphylococcal strain 1 with 25 μg / ml and the staphylococcal strain 2 with 25 μg / ml.



   The α- (4-methyl-1-naphthyl) -diphenylacetyl chloride, m.p. 176-177 C (decomposition) was prepared from the corresponding acid in benzene by heating with thionyl chloride and a trace of pyridine.



     Example 7: Tri- (p-methoxyphenyl) methylpenicillin.



   2.94 g of 6-aminopenicillanic acid were treated with crude tri- (p-methoxyphenyl) acetyl chloride (obtained by heating 5.14 g of the corresponding acid in benzene with thionyl chloride and then removing the solvent and excess reagent in vacuo) according to the method described in Example 4 acylated method described, only the solvent extractions were carried out with butanol instead of ether. 3.44 g of the crude sodium salt of tri (p-methoxyphenyl) methylpenicillin were obtained as a slightly hygroscopic white powder. The test with hydroxylamine showed a purity of approximately 73%.



   The product inhibited Staphylococcus Oxford at a concentration of 5 µg / ml, Staphylococcal strain 1 at 12.5 µg / ml and Staphylococcal strain 2 at 12.5 µg / ml.



     Tri- (p-methoxyphenyl) acetic acid, melting point 212-213 C, was prepared in good yield by condensation of anilinic acid with anisole in the presence of sulfuric acid and acetic acid. b) Diarylalkyl type.



    Example 8: α, α-Diphenylethylpenicillin.



   A solution of 0.49 g of α, α-diphenypropionyl chloride in 6 ml of dry chloroform was added dropwise to a stirred mixture of 0.43 g of 6-aminopenicillanic acid, 12 ml of chloroform and 0.84 ml of triethylamine. The mixture was stirred for 21/2 hours at room temperature and any residues which had not yet dissolved were then filtered off. The filtrate was successively with

 <Desc / Clms Page number 13>

 
 EMI13.1
 

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 c) aryl-dialkyl type.



   Example 13: α, α-Dimethylbenzylpenicillin.



   A solution of 5.2 g of α, α-dimethylphenyl chloride in 40 ml of dry acetone was added dropwise over 15 minutes to a stirred solution of 5.6 g of 6-aminopenicillanic acid and 6.5 g of sodium bicarbonate in 90 ml of water and 50 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 2 1/2 hours, then diluted with 50 ml of water and extracted with three portions of 180 ml each of ether. The aqueous solution extracted in this way was covered with 50 ml of ether and adjusted to a pu value of 2 to 3 by adding n-hydrochloric acid. After separating the layers, the aqueous phase was extracted with two portions of 25 ml each of ether, the pH being adjusted to 2-3 in each case.

   The combined ethereal solutions, which contained the free penicillic acid, were washed twice with 50 ml of water each time and then shaken with enough 8% sodium bicarbonate solution that a neutral aqueous phase (pH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution in each case to give the aqueous phase a pH of 7. Evaporation of the combined aqueous solutions at low temperature and reduced pressure gave the crude sodium salt of α, α-dimethylbenzylpenicillin, which was finally dried in a vacuum desiccator.

   The yield was 9.5 g, it was approximately 77% pure as determined by a colorimetric comparison of hydroxylamine against a benzylpenicillin standard.



   The product inhibited Staphylococcus Oxford at a concentration of 0.025 g / ml, the benzylpenicillin-resistant staphylococcal strain 1 at 25 µg / ml and the benzylpenicillin-resistant staphylococcal strain 2 at 25 µg / ml.



    Example 14: α, α-Diethylbenzylpenicillin.



     5.6 g of 6-aminopenicillanic acid were acylated with 6.0 g of α, α-diethylphenylacetyl chloride, as described in Example 13, to the sodium salt of α, α-diethylbenzylpenicillin (8.3 g, purity approximately 71 ml).
 EMI14.1
    Glml, 7.55 g of 6-aminopenicillanic acid were acylated with 8.35 g of α, α-di-n-propylphenylacetyl chloride as described in Example 13 to give the sodium salt of α, α-di-n-propylbenzylpenicilline (13.0 g, purity about 56vlo).
 EMI14.2
 Game 13, to the sodium salt of a,? -Di-?

  -butylbenzylpenicillin acylated (6.5 g, purity about 49%).
 EMI14.3
 
A solution of 3.8 g of ct, α-dicyclohexylphenylacetyl chloride in 20 ml of dry chloroform was added dropwise to a stirred mixture of 2.6 g of 6-aminopenicillanic acid, 50 ml of chloroform, 5 ml of triethylamine. The mixture was stirred for 2 1/2 hours and a clear solution resulted. It was treated successively with small amounts of 0.2N hydrochloric acid, with vigorous shaking in each case until the aqueous phase had a PEr value of 2. 1.84 g of unchanged 6-aminopenicillanic acid separated out, which were recovered. The acidic layer was then separated and discarded and the chloroform solution was washed with 60 ml of water.

   Sufficient 8% aqueous sodium bicarbonate solution was then added to the chloroform solution so that a neutral emulsion (PH 7) was obtained after vigorous shaking. This was concentrated at low temperature and reduced pressure and the remaining white solid residue was dried in a vacuum desiccator. There were thus obtained 3.57 g of the sodium salt of α, α-dicyclohexylbenzylpenicillin (purity about 16%).



   The product inhibited Staphylococcus Oxford at a concentration of 125 g / ml, the staphylococcal strain 1 with 62.5 g / ml and the staphylococcal strain 2 with 50 n g / ml.



     α, α-Dicyclohexylphenylacetyl chloride, m.p. 95-96 C, 'was obtained by heating the corresponding acid in benzene with thionyl chloride and a trace of pyridine.

 <Desc / Clms Page number 15>

 d) trialkyl type.



     Example 18: tert. Butyl penicillin.



   4.0 g of 6-aminopenicillanic acid (65% pure) was treated as described in Example 1 with 2.5 g of trimethylacetyl chloride. It was so the sodium salt of tert. Butylpenicillin as a yellow hygroscopic
 EMI15.1
 



   A solution of 0.98 g of triethylacetyl chloride in 15 ml of dry chloroform was added dropwise over 5 minutes to a stirred mixture of 1.3 g of 6-aminopenicillanic acid, 30 ml of chloroform, 2.5 ml
Triethylamine added. The mixture was stirred at room temperature for 21/2 hours and then filtered off. The filtrate was washed with 60 ml of 0.2N hydrochloric acid and then with 60 ml of water.



   Sufficient 3% aqueous sodium bicarbonate solution was then added to the chloroform solution that a neutral emulsion (PH 7) was obtained after vigorous shaking. This was concentrated at low temperature and reduced pressure and the residue that remained was dried in a vacuum desiccator. The sodium salt of triethylmethylpenicillin was thus obtained as a white solid (1.84 g). Colorimetric comparison of hydroxylamine against a benzylpenicillin standard showed that it was approximately 57% pure.



   The product inhibited Staphylococcus Oxford at a concentration of 0.05 µg / ml, the benzylpenicillin-resistant Staphylococcenstamml with 12.5 µg / ml and the benzylpenicillin-resistant Staphylococcal strain 2 with 12.5 µg / ml.



    Example 20: Tri-n-propylmethylpenicillin.



     3.89 g of 6-aminopenicillanic acid were acylated with 3.68 g of tripropylacetyl chloride, as described in Example 19, to give the sodium salt of tri-n-propylmethylpenicillin (5.98 g, purity about 60%).



   The product inhibited Staphylococcus Oxford at a concentration of 0.05 µg / ml, the benzylpenicillin-resistant staphylococcal strain 1 with 1.25 μg / ml and the benzylpenicillin-resistant staphylococcal strain 2 with 0.6 µg / ml.



   Example 21: Tri-n-butylmethylpenicillin.



   432 mg of 6-aminopenicillanic acid were acylated with 493 mg of tri-n-butyl acetyl chloride, as described in Example 19, to give the sodium salt of tri-n-butylmethylpenicillin, a colorless, deliquescent solid (628 mg, purity about 380/0).
 EMI15.2
 cocci strain 1 at 2.5 g / ml and staphylococcal strain 2 at 2.5 ug / ml. e) Oxysubstituted diaryl types.
 EMI15.3
 Fenwise to a stirred mixture of 0.43 g of 6-aminopenicillanic acid, 12 ml of chloroform and 0.84 ml of triethylamine added. The mixture was stirred for 2 hours at room temperature and any residues which had not yet dissolved were then filtered off.

   The filtrate was treated successively with small amounts of 0.2N hydrochloric acid, with vigorous shaking in each case until the aqueous phase had a pH of 2 to 3.



   The acidic layer was then separated and discarded and the chloroform solution was washed with 20 ml of water. Sufficient 3% aqueous sodium bicarbonate solution was then added to the chloroform solution so that a neutral emulsion (PH 7) was obtained after vigorous shaking. This was concentrated at low temperature and reduced pressure and the pale yellow solid residue that remained was dried in a vacuum desiccator. In this way, 0.90 g of the crude salt of methoxydiphenylmethylpenicillin were obtained. A colorimetric comparison of hydroxylamine against a benzylpenicillin standard showed it was approximately 50% pure.



   The product inhibited Staphylococcus aureus at a concentration of 0.01 µg / ml, the benzylpenicillin-resistant staphylococcal strain 1 with 5 µg / ml and the benzylpenicillin-resistant staphylococcal strain 2 with 5 µg / ml.



   The α-methoxydiphenylacetyl chloride was prepared as a crude oil by heating α-methoxydiphenylacetic acid with thionyl chloride in benzene and then removing the solvent and excess reagent in vacuo.



    Example 23: α-Ethoxydiphenylmethylpenicillin.



   2.6 g of 6-aminopenicillanic acid were acylated with 3.3 g of a-ethoxydiphenylacetyl chloride, as described in Example 22, to give the sodium salt of a-ethoxydiphenylpenicillin (2.8 g, purity about 59 each).

 <Desc / Clms Page number 16>

 



   The product inhibited Staphylococcus aureus at a concentration of 2.5 μg / ml, the staphylococcal strain 1 with 12.5 μg / ml and the staphylococcal strain 2 with 12.5 μg / ml.



   The a-ethoxydiphenylacetyl chloride was prepared as a crude oil by heating a-ethoxydiphenylacetic acid with thionyl chloride in benzene, after which the solvent and the excess of the reagent are removed in vacuo.
 EMI16.1
 Fenwise added to a stirred solution of 3.5 g of 6-aminopenicillanic acid and 4 g of sodium bicarbonate in 48 ml of water and 28 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 2 hours, then diluted with 60 ml of water and extracted with three portions of 100 ml each of ether. The aqueous solution extracted in this way was covered with 30 ml of ether and adjusted to a pH of 2 by adding n hydrochloric acid. After separating the layers, the aqueous phase was extracted with two portions of 15 ml each of ether.

   The combined ether solutions, which contained the free penicillic acid, were washed with 50 ml of water and then shaken with enough 8% sodium bicarbonate solution that a neutral aqueous phase (pH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution in each case to give the aqueous phase a pH of 7.



  The combined aqueous solutions were washed with 15 ml of ether and evaporated at low temperature and reduced pressure. The crude sodium salt of et-isopropoxydiphenylmethylpenicillin was thus obtained as a white solid, which was finally dried in a vacuum desiccator. The yield was 2.66 g, it was approximately 53% pure.



   The product inhibited Staphylococcus Oxford in a concentration of 0.6 μg / ml, the staphylococcal strain 1 with 12.5 μg / ml and the staphylococcal strain 2 with 12.5 μg / ml.



   The α-isopropoxydiphenylacetyl chloride was prepared as a crude oil by heating α-isopropoxydiphenylacetic acid with thionyl chloride in benzene and then removing the solvent and excess reagent in vacuo.



    Example 25: ex-butoxydiphenylmethylpenicillin.



   0.43 g of 6-aminopenicillanic acid were treated with Iga-butoxydiphenylacetyl chloride, as in Example 22
 EMI16.2
   44gO).



   The product inhibited Staphylococcus aureus in a concentration of 1.26 μg / ml, the Staphylococcal strain 1 with 2.5 μg. t g / ml and the staphylococcal strain 2 with 2.5 jug / ml.



   The butoxydiphenylacetyl chloride was prepared as a crude oil by heating a-butoxydiphenylacetic acid with thionyl chloride in benzene and then removing the solvent and excess reagent in vacuo. f) Thio-substituted diaryl types.
 EMI16.3
 added wise over 10 min to a stirred solution of 5.4 g of 6-aminopenicillanic acid and 6.3 g of sodium bicarbonate in 90 ml of water and 60 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 2 1/2 hours, then diluted with 50 ml of water and extracted with two portions of 100 ml each of ether. The aqueous solution extracted in this way was covered with 50 ml of ether and adjusted to a pH of 2 by adding n-hydrochloric acid.

   After separating the layers, the aqueous phase was extracted with two further portions of 25 ml each of ether. The combined ether solutions, which contained the free penicillic acid, were washed with 50 ml of water and then shaken with enough 80% sodium bicarbonate solution that a neutral aqueous phase (pH 7) was obtained.



  The layers were separated. and the ether phase was extracted with two portions of 5 ml of water each time, sufficient bicarbonate solution being added to give the aqueous phase a pH of 7. The combined aqueous solutions were washed with 25 ml of ether and then evaporated at low temperature and reduced pressure. The crude sodium salt of α-ethylthiodiphenylmethylpenicillin was thus obtained as a pale yellow solid, which was finally dried in a vacuum desiccator. The yield was 8.67 g, it was approximately 84% pure as indicated by a colorimetric comparison of hydroxylamine against a benzylpenicillin standard.



   The product inhibited Staphylococcus Oxford at a concentration of 0.025 µg / ml, the benzylpenicillin-resistant staphylococcal strain 1 at 1.25 µg / ml and the benzylpenicillin-resistant staphylococcal strain 2 at 2.5 µg / ml.

 <Desc / Clms Page number 17>

 



    The # -ethylthiodiphenylacetyl chloride was prepared as a crude oil by heating the appropriate acid with thionyl chloride in benzene and removing the solvent and excess reagent in vacuo. The α-ethylthiodiphenylacetic acid itself, mp = 131-132 ° C., was prepared by compensating benzilic acid with ethyl mercaptan according to the method described by Bakenbus and Brower (J. Amer. Chem. Soc., I 77 [1955], p. 579).



  Example 27: α-n-Propylthiodiphenylmethylphenicillin.



  6.25 g of 6-aminopenicillanic acid was acylated with 8.8 g of α-n-propylthiodiphenylacetyl chloride as described in Example 26 to give the sodium salt of α-n-propylthiodiphenylmethylpenicillin, a white powder (8.36 g, purity approximately 65 %).



  The product inhibited Staphylococcus Oxford in a concentration of 0.6 blglrnl, the staphylococcus strain 1 with 2.5 J, Lg / ml and the staphylococcus strain 2 with 1.25 g / ml. The an-propylthiodiphenylacety1chloride was produced as a crude oil by Warming the corresponding acid with thionyl chloride in benzene and removing the solvent and the excess of the
 EMI17.1
 
A solution of 0.64 g of α-n-butylthiodiphenylacetyl chloride in 6 ml of dry chloroform was added dropwise to a stirred mixture of 0.43 g of 6-aminopenicillanic acid, 12 ml of chloroform, 0.84 ml of triethylamine. The mixture was stirred at room temperature for 2½ hours and then filtered.

   The filtrate was treated successively with small amounts of 0.2N hydrochloric acid, with vigorous shaking in each case until the aqueous phase had a pH of 2.



   The acidic layer was then separated and discarded and the chloroform solution was washed with 15 ml of water. Sufficient 3% strength aqueous sodium bicarbonate solution was then added to the chloroform solution that, after vigorous shaking, a neutral emulsion (PH 7) was obtained. This was concentrated at low temperature and reduced pressure and the remaining white solid residue was dried in a vacuum desiccator. There was thus obtained 0.62 g of the sodium salt of a-n-butylthiodiphenylmethylpenicillin (purity about 46%).



   The product inhibited Staphylococcus aureus in a concentration of 1.25 I, Lg / ml, the staphylococcal strain 1 with 2.5 J. Lg / ml and the staphylococcal strain 2 with 2.5 g / ml.
 EMI17.2
 cocci stem 1 with 5 g / ml and the staphylococcal stem 2 with 5 pg / ml.



   The α-isopropylthiodiphenylacetyl chloride was prepared as a crude oil by heating the appropriate acid with thionyl chloride in benzene and removing the solvent and excess reagent in vacuo. The α-isopropylthiodiphenylacetic acid itself, m.p. 130-132 C, was prepared by compensating benzilic acid with isopropyl mercaptan according to the method described by Barkenbus and Brower (J. Amer.



  Chem. Soc., 77 [1955], p. 579).



     Example 30:? -Sec. Butylthiodiphenylmethylpenicillin.



     5.4 g of 6-aminopenicillanic acid were acylated with 9.6 g (x-sec. Butylthiodiphenylacetyl chloride, as described in Example 26, to the sodium salt of a-sec. Butylthiodiphenylmethylpenicillin, a pale yellow solid (7.87 g, purity about 590 / 0).



   The product inhibited Staphylococcus Oxford at a concentration of 0.6 μLg / ml, the staphylococcal strain 1 with 5 g / ml and the staphylococcal strain 2 with 5 μLfg / ml.



   The a-sec. Butylthiodiphenylacetyl chloride was prepared as a crude oil by heating the appropriate acid with thionyl chloride in benzene and removing the solvent and excess reagent in vacuo. The (x-sec. Butylthiodiphenylacetic acid itself, mp = 95-97 ° C., was prepared by compensating benzilic acid with sec. Butyl mercaptan according to the method described by Barkenbus and Brower (J.



  Amer. Chem. Soc. 77 [1955], p. 579).

 <Desc / Clms Page number 18>

 



    Example 31: a. -Benzylthiodiphenylmethylpenicillin.



   4.95 g of 6-aminopenicillanic acid were acylated with 8.85 g of α-benzylthiodiphenylacetyl chloride, as described in Example 26, to give the sodium salt of (x-benzylthiodiphenylmethylpenicillin (7.53 g, purity about 68%).



   The product inhibited Staphylococcus Oxford at a concentration of 0.125 μl / ml, staphylococcal strain 1 at 0.5 μlg / ml and staphylococcal strain 2 at 0.6 μg / ml.



   The α-benzylthiodiphenylacetyl chloride was prepared as a crude oil by heating the corresponding acid, which itself was prepared by the method of Barkenbus and Brower (J. Amer. Chem. Soc., 77 [1955], p. 579) with thionyl chloride in benzene and subsequent removal of the solvent and the solvent and the excess of the reagent in vacuo.
 EMI18.1
 game 28 described, acylated to the sodium salt of α-phenylthiodiphenylmethylpenicillin (0.34 g, purity about 49%).



   The product inhibited Staphylococcus aureus in a concentration of 0.5 Mglml, the staphylococcal strain 1 with 1.25 μg / ml and the staphylococcal strain 2 with 1.25 g / ml.



   The a-phenylthiodiphenylacetyl chloride, m.p. 67-69 ° C., was prepared by heating the corresponding acid in benzene, with thionyl chloride and a trace of pyridine, removing the solvent and the excess of the reagent in vacuo and crystallizing the residue from petroleum ether.
 EMI18.2
 
33: <x-p-Tolylthiodiphenylmethylpenicillin.spiel26 described, acylated to the sodium salt of (x-p-tolyldiphenylmethylpenicillin (7.5 g, purity about 58%).



   The product inhibited Staphylococcus Oxford at a concentration of 0.6 lg / ml, the staphylococcal strain 1 with 1.25 μg / ml and the staphylococcal strain 2 with 1.25 pg / ml.



   Slightly contaminated a-p -tolylthlodiphenylacetyl chloride was prepared by heating the appropriate acid in benzene with thionyl chloride and some pyridine. After removing the solvent and excess reagent in vacuo, the remaining oil was scraped off to initiate crystallization and the resulting colorless solid, mp = 70-74 ° C., was used without further purification. g) Oxy-substituted dialkyl types.



    Example 34: α-Phenoxyisopropylpenicillin.



   A solution of 5.77 g of a-phenoxyisobutyryl chloride in 40 ml of dry acetone was added dropwise over 15 minutes to a stirred solution of 5.63 g of 6-aminopenicillanic acid and 6.6 g of sodium bicarbonate in 90 ml of water and 50 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 2 hours, then diluted with 50 ml of water and finally washed with 2 portions of 90 ml each of ether. The aqueous phase was brought to a pH of 2 by adding 57 ml of n-hydrochloric acid and the released penicillic acid was extracted into three times 25 ml of ether. These ether extracts were washed twice with 20 ml of water and then shaken with enough sodium bicarbonate solution that a neutral aqueous phase (pH 7) was obtained.

   The layers were separated and the ether phase was washed twice with 5 ml of water, adding enough bicarbonate solution to each time that an aqueous phase with a pH of 7 was obtained. Evaporation of the combined aqueous solutions at low temperature and reduced pressure gave the crude salt of 6 - (α-phenoxy-α-methylpropionamino) -penicillanic acid as a white solid which was finally dried in a vacuum desiccator. The yield was 10.6 g. Colorimetric comparison of hydroxylamine against a benzylpenicillin standard indicated that the product was approximately 78% pure.
 EMI18.3
 Ber. 46 [1913], p. 1347.



   Example 35: A solution of 0.72 g of a-phenoxyisobutyric acid and 0.55 ml of triethylamine in 8 ml of dry acetone was stirred and cooled to 0.degree. A solution of 0.51 ml sec. Butyl chloroformate in 8 ml of dry acetone was added dropwise over 5 minutes, after which the mixture was stirred at 0 ° -50 ° C. for 1 hour. A solution of 0.86 g of 6-aminopenicillanic acid in 16 ml of 0.5N sodium bicarbonate solution was then added to the suspension and the resulting clear solution was stirred at room temperature for 2 hours. It was then washed twice with 15 ml of ether and the

 <Desc / Clms Page number 19>

 Ether was discarded.

   The aqueous phase was then acidified to a pH of 2.5 and the 6- (a-phenoxy- (x-methylpropionamide) -penicillanic acid, as described in Example 34, isolated. The yield of sodium salt was 1.29 g, its purity about 65%.



   Example 36: 0.1 mol of isobutyl chloroformate were added to a cooled solution of
 EMI19.1
 Water was added to the above acylation mixture and the mixture was stirred in the cold for about 1 hour. After adding a saturated solution of sodium bicarbonate, the alkaline reaction mixture was extracted with ether and the ethereal extract was discarded. Now the aqueous layer was covered with methyl isobutyl ketone, acidified with 6N hydrochloric acid to a pH of about 2 and then extracted twice with methyl isobutyl ketone. The combined extracts, which contained the 6- (a-phenoxy-ci-methylpropionamido) -penicillanic acid, were washed with water, dried over anhydrous sodium sulfate and filtered.

   The potassium salt of the above penicillanic acid was precipitated by adding 40 ml of a 40% strength butanol solution of potassium 2-ethylbexanoate. After treatment with ether, this potassium salt was dried in vacuo over phosphorus pentoxide. It is a water-soluble powder, weighing 20.8 g (0.05 mole), melts at 232-234 ° C. with decomposition and has a B-lactam structure as shown by infrared analysis. It inhibits Staphylococcus aureus Smith in one
 EMI19.2
 40 ml of dry acetone was added dropwise over 15 minutes to a stirred solution of 5.63 g
6-aminopenicillanic acid and 6.6 g of sodium bicarbonate in 90 ml of water and 50 ml of acetone were added.

   After the addition was complete, the mixture was stirred at room temperature for 2 hours, then diluted with 50 ml of water and extracted with two 90 ml portions of ether. The aqueous phase was brought to a pH value of 2 by adding 57 ml of n-hydrochloric acid and the penicillic acid released in this way was extracted into three times 25 ml of ether.



   These ether extracts were washed twice with 20 ml of water and then shaken with enough 8% sodium bicarbonate solution that a neutral aqueous phase (PH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution in each case to give the aqueous phase a pH of 7. Evaporation of the combined aqueous solutions at low temperature and reduced pressure gave the crude sodium salt of 6 - [α- (2,4-dichlorophenoxy) -α -benzyl-n-butylamidol- penicillanic acid as a white solid, which was finally converted into dried in a vacuum desiccator.



   The product inhibited Staphylococcus aureus Smith at a concentration of 0.001% by weight.



     Example 38: 0.1 mol isobutyl chloroformate was added to a cooled (0-3 C) solution of 0.1 mol of α- (4-hydroxyphenoxy) -α-methylisovaleric acid and 0.1 mol of triethylamine in 250 ml of acetone and 500 ml of dioxane was added and the resulting mixture was stirred in the cold for about 30 minutes. The mixture was then treated with a cooled solution of 0.1 mol of 6-aminopenicillanic acid and 0.1 mol of triethylamine in 500 ml of water and stirred in the cold for a further hour. The pH of the reaction mixture was adjusted to approximately 8 by the addition of a saturated solution of sodium bicarbonate solution and the mixture was then extracted with ether.

   Now the aqueous layer was covered with methyl isobutyl ketone, acidified with hydrochloric acid to a pH of about 2 and then extracted twice with methyl isobutyl ketone. The combined extracts containing the 6 [α- (4-hydroxyphenoxy) -α-methylisovalerianamido] penicillanic acid were washed with water, dried over anhydrous sodium sulfate and filtered. The potassium salt of the above penicillanic acid was precipitated by adding 40 ml of a sufficient butanol solution of potassium 2-ethylhexanoate. After treatment with ether, this potassium salt was dried in vacuo over phosphorus pentoxide. It is a water-soluble powder which inhibited Staphylococcus aureus Smith at a concentration of 0.001% by weight.



     At pie 1 3 9: 0.1 mole isobutyl chloroformate was added to a chilled (40C) solution of 0.1 mole of α- (2,4-dimethoxyphenoxy) -α-methyl-n-valeric acid and 0.1 mole of triethylamine in 250 ml
 EMI19.3
 

 <Desc / Clms Page number 20>

 Stirred in the cold for about 1 hour. The reaction mixture was then made slightly basic (PH 8) by adding a saturated solution of sodium bicarbonate and then extracted with ether. Now the aqueous layer was covered with methyl isobutyl ketone, acidified with 6N hydrochloric acid to a pH of about 2 and then extracted twice with methyl isobutyl ketone.

   The combined extracts containing the 6 - [α- (2,4-dimethoxyphenoxy) -α-methyl-n-valerylamido] penicillanic acid were washed with water, dried over anhydrous sodium sulfate and filtered. The potassium salt of the above penicillanic acid was precipitated by adding 40 ml of a 40% butanol solution of potassium 2-ethylhexanoate. After treatment with ether, this potassium salt was dried in vacuo over phosphorus pentoxide. It is a water-soluble powder which inhibits the growth of Staphylococcus aureus Smith in a concentration of 0.001% by weight.



     Example 40: A solution of 9.6 g <x-phenoxy-a-benzylphenylacetyl chloride in 40 ml dry acetone was added dropwise over 15 min to a stirred solution of 5.3 g 6-aminopenicillanic acid and 6.6 g sodium bicarbonate in 90 ml water and 50 ml of acetone added. After the addition was complete, the mixture was stirred at room temperature for 2 hours, then diluted with 50 ml of water and washed with two portions of 90 ml of ether each. The aqueous solution extracted in this way was brought to a pH value of 2 with 57 ml of n-hydrochloric acid and the released penicillic acid was extracted with three times 25 ml of ether.

   These ether extracts were washed twice with 20 ml of water and then shaken with enough 8% sodium bicarbonate solution that a neutral aqueous phase (PH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution in each case to give the aqueous phase a pH of 7. Evaporation of the combined aqueous solutions at low temperature and reduced pressure gave the crude sodium salt of 6 - (α-phenoxy-α-benzylphenylacetamide) - penicillanic acid as a white dye, which was finally dried in a vacuum desiccator.



   The product inhibited Staphylococcus aureus Smith at a concentration of 0.001% by weight.



   Example 41: In the processes described in Examples 34-36, the acid was replaced by the following acids:
 EMI20.1
 -Phenoxy- <x -benzylpropionic acid, 6 (α-phenoxy-α-benzylphenylbutyramido) -penicillanic acid, 6 [α- (4-nitrophenoxy) -α-methyl-n-butyramido] -penicillanic acid,
 EMI20.2
 - (4-Bromophenoxy) -a-phenyl-n -valerianamido] -penicillanic acid, 6 [α- (3-methoxyphenoxy) -α-methyl-decanamido] -penicillanic acid, 6 - [α- (2-iodophenoxy) -α-phenylpropionamido] -penicillanic acid, 6 [α- (3-diethylaminophenoxy) -α-methyl-n-butyramido] -penicillanic acid,

 <Desc / Clms Page number 21>

 
 EMI21.1
 h) Thio-substituted dialkyl type.



    Example 42: α-Phenylthioisopropylpenicillin.



   A solution of 6.24 g of α-phenylthioisobutyryl chloride in 30 ml of dry acetone was added dropwise over 10 minutes to a stirred solution of 5.63 g of 6-aminopenicllanic acid and 6.55 g of sodium bicarbonate in 90 ml of water and 60 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 2 hours, then extracted twice with 90 ml of ether. The aqueous solution extracted in this way was covered with 50 ml of ether and adjusted to a pH of 2 by adding n-hydrochloric acid. After separating the layers, the aqueous phase was extracted with two portions of 25 ml each of ether.

   The combined ether solutions, which contained the free penicillic acid, were washed with 50 ml of water and then shaken with enough sodium bicarbonate solution that a neutral aqueous phase (pH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution in each case to give the aqueous phase a pH of 7. The combined aqueous extracts were washed with 30 ml of ether and evaporated at low temperature and reduced pressure. The residue was dried in a vacuum desiccator. 8.7 g of the crude salt of α-phenylthioisopropylpenicillin were thus obtained as a white solid.

   Colorimetric comparison of hydroxylamine against a benzylpenicillin standard showed that it was approximately 60% pure.



   The product inhibited Staphylococcus Oxford at a concentration of 0.012 g / ml, the benzylpenicillin-resistant staphylococcal strain 1 with 26 g / ml and the benzylpenicillin-resistant staphylococcal strain 2 with 12.5 g / ml.



    Example 43: α-Ethyl-α-phenylthiopropylpeniclin,
6.49 g of 6-aminopenicillanic acid were mixed with 7.28 g of a-ethyl-a-phenylthiobutyryl chloride according to the
 EMI21.2
 



   The product inhibited Staphylococcus Oxford in a concentration of 0.1 g / ml, the staphylococcal strain 1 with 5 g / ml and the staphylococcal strain 2 with 2.5 f. ! g / ml.



     α-Ethyl-α-phenylthiobutyryl chloride, b.p. 86 C, was converted from the corresponding acid and thionyl
 EMI21.3
    s pie 1 44: a-Propyl-a-phenylthiobutylpenicillin. 4, 32 g of 6-aminopenicillanic acid were mixed with 5.42 g of ct-propyl-a-phenylthiovaleryl chloride according to the method of Example 42 to give the sodium salt of α-propyl-α- phenylthiobutylpenicillin acylated (8.6 g, approximately 52% pure).
 EMI21.4
 tronlauge produced. j) Heterocyclic diaryl type.



    Example 45: α, α-Diphenyl-2-thienylmethylpenicillin.



   A solution of 6.3 g of α-2-thienyldiphenylacetyl chloride in 40 ml of dry acetone was slowly added to a stirred solution of 4.33 g of 6-aminopenicillanic acid and 5 g of sodium bicarbonate in 100 ml of water and 60 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 3 hours and then filtered. The filtrate was extracted with twice 100 ml of ether and the extracts were discarded. The aqueous phase was covered with 60 ml of ether and brought to a pH of 3 by adding n-hydrochloric acid. After separating the layers, the aqueous phase was extracted with two further portions of 25 ml each of ether.

   The combined ether solutions, which contained the free penicillic acid, were washed with 50 ml of water and then with enough 8% sodium bicillate

 <Desc / Clms Page number 22>

 shaken carbonate solution that a neutral aqueous phase (PH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution to give the aqueous phase a pH of 7. The combined aqueous extracts were washed with ether and then evaporated at low temperature and reduced pressure. Further drying in a vacuum desiccator gave the crude salt of α-2-thienyldiphenylmethylpenicillin as an off-white solid (6.94 g).

   Colorimetric comparison of hydroxylamine against a benzylpenicillin standard showed that it was approximately 51% pure.



   The product inhibited Staphylococcus aureus at a concentration of 0.5 / lg / ml, the benzylpenicillin-resistant staphylococcal strain 1 with 0.6 μg / ml and the benzylpenicillin-resistant staphylococcal strain 2 with 0.6 / lg / ml.



     (x-2-Thienyldiphenylacetyl chloride, m.p. 76-79 C, was prepared by heating the corresponding acid in benzene with thionyl chloride and a trace of pyridine. k) 1-Arylcycloalkyl type.



   Example 46: 1-Phenylcyclopentylpenicillin.



   A solution of 6.0 g of 1-phenylcyclopentane-1-carbonyl chloride in 30 ml of dry acetone was gradually added to a stirred solution of 5.63 g of 6-aminopenicillanic acid and 6.55 g of sodium bicarbonate in 90 ml of water and 60 ml of acetone . After the addition was complete, the mixture was stirred at room temperature for 2 1/2 hours, then extracted twice with 90 ml of ether. The aqueous solution was covered with 50 ml of ether and adjusted to a pH of 2 by adding n-hydrochloric acid.



  After separating the layers, the aqueous phase was extracted with two portions of 25 ml each of ether.



  The combined ether solutions, which contained the free penicillic acid, were washed with 50 ml of water and then shaken with enough 8% strength sodium bicarbonate solution that a neutral aqueous phase (pH 7) was obtained. The layers were separated and the ether phase became two
 EMI22.1
 were washed with 15 ml of ether and then evaporated at low temperature and reduced pressure. The residue was finally dried in a vacuum desiccator. The crude salt of 1-phenylcyclopentylpenicillin was thus obtained as a light brown, hygroscopic solid (9.78 g). A colorimetric comparison of hydroxylamine against a benzylpenicillin standard showed it was approximately 74% pure.



   The product inhibited Staphylococcus Oxford at a concentration of 0.005 flg! ml, the benzylpenicillin-resistant staphylococcal strain 1 with 12.5 µg / ml and the benzylpenicillin-resistant staphylococcal strain 2 with 12.5 µg / ml.



    Example 47: 1-Phenylcyclohexylpenicillin.



     5.63 g of 6-aminopenicillanic acid were mixed with 6.4 g of 1-phenylcyclohexane-1-carbonyl chloride, as described in Example 46, to give the crude salt of 1-phenylcyclohexylpenicillin in the form of a white solid (7.8 g, approximately 69% pure) acylated.
 EMI22.2
 
A solution of 0.67 g of 9-p-methoxyphenylfluorene-9-carbonyl chloride in 8 ml of dry chloroform was added dropwise to a stirred mixture of 0.43 g of 6-aminopenicillanic acid, 12 ml of chloroform and 0.84 ml of triethylamine. The mixture was stirred for a further 2½ hours and then filtered. The filtrate was treated successively with small amounts of 0.2N hydrochloric acid, with vigorous shaking in each case until the aqueous phase had a pH of 3.



   The acidic layer was then separated and discarded and the chloroform solution was washed with 20 ml of water. Sufficient 30% aqueous sodium bicarbonate solution was then added to the chloroform solution that a neutral emulsion (PH 7) was obtained after vigorous shaking. This was concentrated at low temperature and reduced pressure and the yellow solid residue that remained was dried in a vacuum desiccator. 0.83 g of the sodium salt of 9-p-methoxyphenyl- 9-fluorenylpenicillin were thus obtained. Colorimetric comparison of hydroxylamine versus a benzylpenicillin standard showed it was approximately 53% pure.



   The product inhibited Staphylococcus aureus at a concentration of 1.25 years! g / ml, the benzylpenicillin-resistant staphylococcal strain 1 with 2.5 / lg! ml and the benzylpenicillin-resistant staphylococcal strain 2 with 2, 5! mol.

 <Desc / Clms Page number 23>

 



     9-p-Methoxyphenylfluorene-9-carbonyl chloride, m.p. 169-171 C, was prepared by heating 9-p-methoxyphenylfluorene-9-carboxylic acid in benzene with thionyl chloride and a trace of pyridine.



    Example 49: 9-Methoxy-9-fluorenylpenicillin.



     3.81 g of 9-methoxyfluorene-9-carboxylic acid, 1.1 ml of thionyl chloride and 10 drops of pyridine in 15 ml of benzene were refluxed for 75 minutes and then evaporated in vacuo. The remaining impure waxy acid chloride was dissolved in 20 ml of dry acetone and added dropwise to a stirred solution of 3.42 g of 6-aminopenicillanic acid and 4 g of sodium bicarbonate in 48 ml of water and 20 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 3 1/2 hours, then diluted with 50 ml of water and extracted twice with 50 ml of ether. The aqueous solution was covered with 50 ml of ether and adjusted to a pH of 2.5 to 3 by adding n-hydrochloric acid.



  After separating the layers, the aqueous phase was extracted with two further portions of 25 ml each of ether. The combined ether solutions, which contained the free penicillic acid, were washed with water and then shaken with enough pure sodium bicarbonate solution that a neutral aqueous phase (PH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution to give the aqueous phase a pH of 7.



   Evaporation of the combined aqueous solutions at low temperature and reduced pressure gave the crude salt of 9-methoxy-9-fluorenylpenicillin, which was finally dried in a vacuum desiccator. Yield 3.2g, about 53% pure.



   The product inhibited Staphylococcus Oxford in a concentration of 0.25 μg / ml, the Staphylococcus strain 1 with 25 μg / ml and the Staphylococcus 2 with 12.5 μg / ml.



   Example50: 9-ethoxy-9-fluorenylpenicillin,
 EMI23.1
 were converted to the crude acid chloride and the latter was obtained as a white solid (5.63 g, approx.), sodium salt of 9-ethoxy-9-fluorenylpenicillin
53% pure).



   The product inhibited Staphylococcus Oxford in a concentration of 1.25 J. lg (ml. M) Various types.



     Example 51: Trichloromethylpenicillin.



     3.0 g of 6-aminopenicillanic acid (65% pure) were reacted with 2.8 g of trichloroacetyl chloride, as described in Example 1, to give the sodium salt of trichloromethylpenicillin, a pale yellow hygroscopic solid (1.7 g, approximately 38% pure).



   The product inhibited Staphylococcus aureus in a concentration of 1.25 μg / ml.



    Example 52: α-Chlorine-α-cyclohexylbenzylpenicillin.



   A solution of 0.63 g of cx-chloro-cx-cyclohexyl-cx-phenylacetyl chloride in 5 ml of dry chloroform was added dropwise to a stirred mixture of 0.43 g of 6-aminopenicillanic acid, 12 ml of chloroform and 0.84 ml of triethylamine. The mixture was stirred for 2½ hours. The resulting solution was treated successively with small amounts of 0.2N hydrochloric acid, with vigorous shaking in each case until the aqueous phase had a pH of 2 to 3.



   The acidic layer was then separated and discarded and the chloroform solution was washed with 20 ml of water. Sufficient 3% aqueous sodium bicarbonate solution was then added to the chloroform solution so that a neutral emulsion (PH 7) was obtained after vigorous shaking. This was concentrated at low temperature and reduced pressure and the yellow solid residue that remained was dried in a vacuum desiccator. This gave 0.84 g of the sodium salt of α-chloro-α-cyclohexylbenzylpenicillin (approximately 55% pure).



   The product inhibited Staphylococcus aureus in a concentration of 0.25 μl / ml. Staphylococcus Oxford with 0.025 g / ml, the benzylpenicillin-resistant staphylococcal strain 1 with 12.5 g / ml and the benzylpenicillin-resistant staphylococcal strain 2 with 25 μg / ml.



    Example 53: α-Bromo-α-ethylpropylpenicillin.
 EMI23.2
 added over 5 min to a stirred solution of 5.85 g of 6-aminopenicillanic acid and 6.8 g of sodium bicarbonate in 80 ml of water and 50 ml of acetone. After the addition was complete, the mixture was stirred at room temperature for 3 hours and then diluted with 40 ml of water and extracted twice with 80 ml of ether.



  This aqueous solution was covered with 50 ml of ether and brought to a pH value of 2 by adding n-hydrochloric acid. After separating the layers, the aqueous phase was extracted with two further portions of 25 ml each of ether. The combined ethereal solutions which form the free penicillic acid

 <Desc / Clms Page number 24>

 were washed with 50 ml of water and then shaken with enough sodium bicarbonate solution that a neutral aqueous phase (PH 7) was obtained. The layers were separated and the ether phase was extracted with two 5 ml portions of water, adding enough bicarbonate solution to give the aqueous phase a pH of 7. The combined aqueous extracts were washed with 15 ml of ether and then evaporated at low temperature and reduced pressure.

   Further drying in a vacuum desiccator gave the crude salt of ex-bromo-o.-ethylpropylpenicillin as a white solid. Yield 9.26g (approximately 68% pure).



   The product inhibited Staphylococcus Oxford at a concentration of 0.12 μg / ml, the staphylococcal strain 1 with 25 / μg / ml and the staphylococcal strain 2 with 12.5 g / ml.



   The compounds according to the invention can be used in a mixture with suitable pharmaceutical carriers in various forms for therapeutic use. According to the invention, a mixture can therefore also be provided which consists of a pharmaceutical carrier and a compound of the general formula (II).



    PATENT CLAIMS:
1. Method zur'Herstellung of new penicillin derivatives and their therapeutically useful salts, characterized in that 6-aminopenicillanic acid with an acid chloride, bromide, anhydride or with a mixed anhydride with another acid, of a trisubstituted acetic acid of the general formula
 EMI24.1
 wherein Rl, R2 and R3 each have a maximum of 24 carbon atoms.

   Rl and R2 are identical or different and each represent a halogen atom or an optionally substituted alkyl, alkenyl, aryl, aralkyl, cycloalkyl or heterocyclic group, or R1 and R2 together with the carbon atom to which they are bonded are, form an aromatic, cycloaliphatic or heterocyclic ring system and R3 is one of the groups listed above under Rl and R2 or the group OR4 or SR4, where R is an alkyl, alkenyl, aryl, aralkyl or cycloalkyl group, with a Temperature from about 0 to 300C and at a pH of about 6 to 9 to compounds of the general formula
 EMI24.2
 in which Rl - R3 have the meaning given above, is reacted.

 

Claims (1)

2. The method according to claim 1, characterized in that Rl denotes a substituted aromatic ring, at least one of the substituents is a hydroxyl group and R2 and R3 are identical or different and the same group as R1 or an optionally substituted alkyl, aryl or aralkyl group mean. 3. The method according to claim 1, characterized in that R is a substituted or unsubstituted benzene ring, R2 is an alkyl group or an alkoxy group, both of which contain 1-6 carbon atoms and R3 is one of the groups specified under R1 and R2. 4. The method according to claim 1, characterized in that a trisubstituted acetic acid of the general formula <Desc / Clms Page number 25> EMI25.1 is used, wherein X, Y and Z are the same or different and are each hydrogen, nitrogen or a halogen atom or an amino, alkylamino, dialkylamino, acylamino, alkanoylamino, alkyl, alkoxy, hydroxy, sulfamyl, Benzyl, cyclohexyl, cyclopentyl or trifluoromethyl group and A and B are identical or different and each represent an alkyl, phenyl or phenylalkyl group.