BE670219A - - Google Patents

Description

  

   <Desc / Clms Page number 1>
 



    "New penicillins".



   The present invention relates to certain novel synthetic penicillins and their production methods, as well as to certain intermediates used in these methods.



   The invention relates in particular to adamantyl- (1) -penicillins substituted in position 3, of general formula (I);

 <Desc / Clms Page number 2>

 
 EMI2.1
 wherein each of R1 and R8 denotes a hydrogen atom or a lower alkyl radical containing 1 to 4 carbon atoms, and the invention also relates to the pharmaceutically acceptable salts with the readily hydrolyzable esters of these penicillins .



   In addition to the aforesaid free penicillins of formula (1) and their salts and esters, the present invention also encompasses penicillins in the form of their ampho-ions, pharmaceutical compositions and unit dosages containing these. penicillins.



   The penicillins of the present invention are broad spectrum antibiotics and are characterized in that they are resistant to the enzyme penicillinase, hereinafter referred to as "penase", and that they additionally possess activity. antibacterial which compares favorably with the activity of other penicillins resistant to penase.



   The compounds of the present invention are non-toxic penicillins and are well tolerated when used. They possess an acid stability which corresponds to that of penicillin G and are, therefore, suitably administered in flexible pharmaceutical dosage forms which are known and available. preferred for penicillin G.



   Penicillins resistant to penase are known,
 EMI2.2
 for example methicillin (2,5-dimethoxyphenylpenillin), which is widely used in the clinical treatment of infectious diseases caused by staphylococci, the resistance of which is

 <Desc / Clms Page number 3>

 this to penicillin is due to the formation of this anzyme.



   The only advantage of the previous methicillin is that it is resistant to penase because, in terms of its antibacterial activity, it is only one hundredth of that of penicillins G or V. These last penicillins are, therefore , those which are preferred and utilized by the practitioner in the routine treatment of patients suffering from infectious diseases, and their administration is only stopped if it appears to be unsuccessful, indicating that the disease may be caused. by microorganism resistant to penicillin and that methicillin or a corresponding penicillin should be prescribed.



   In such cases, a period of value processing is lost, which also occurs in addition also if, in order to choose the most advantageous penicillin, a prior determination of the susceptibility of the pathogenic microorganism vis -with various penicillins is carried out.



   An object of the present invention is to provide penicillins in which the very high antibacterial activity associated with, for example, penicillins G and V, is combined with the resistance to methicillin penase, so that the new penicillins can be advantageously applied in the routine treatment of infectious diseases without wasting treatment time.



   By combining certain adamantha- (1) -carboxylic acids substituted in position 3, with 6-amino-penicillinic acid, penicillins are obtained which, besides the advantage mentioned above, have been found to be particularly active against bacteria not taking the gram, among which, as is known, many pathogenic microorganisms less susceptible to previously known penicillins develop and some of which are penase producers.

 <Desc / Clms Page number 4>

 



   In Tables I and II below, the activity of one of the penicillins of the present invention has been illustrated in greater detail in the form of the results obtained in comparative serial dilution tests.



   In the tables, the activity of the penicillin involved is expressed by the negative logarithm of its concentration which produces a 50% inhibition of growth of the predicted microorganism.



   Apart from the exceptions marked with an asterisk in the tables, the culture medium used in the test was a solution of casein hydrolyzate / yeast extract with the addition of L-cysteine.



   The term WG 832 used in the tables denotes 3-amino-adamantyl - (1) -penicillin.



   TABLE 1
 EMI4.1
 
<tb> Penicillin <SEP> Organism <SEP> of <SEP> test
<tb> Staphylococcus <SEP> aureus, <SEP> Staphylococcus <SEP> aureus,
<tb>
<tb> no <SEP> producer <SEP> of <SEP> producer <SEP> of <SEP> penase
<tb>
 
 EMI4.2
 penase ¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯¯
 EMI4.3
 
<tb> Methicillin <SEP> 6.2 <SEP> 6.0X <SEP> 6.0
<tb>
 
 EMI4.4
 WG 832 6, 9 6.0 6.1
 EMI4.5
 
<tb> x <SEP> The <SEP> substrate <SEP> containing <SEP> 50% <SEP> of <SEP> serum.
<tb>
 



   It appears from Table I that 3-amino-adamantyl- (1) - penicillin is as active as methicilin when a penase-producing strain of Staphylococcus aureus constitutes the test organism, while it is also six times. active than methicillin when tested on a non-penase-producing strain of the same organism. It is further found that this higher activity is not affected by the serum present in the culture medium.



   The beneficial high activity of 3-amino-adamantyl- (1) -penicillin against non-gram-taking bacteria is illustrated in the following Table II.

 <Desc / Clms Page number 5>

 



  TABLE II
 EMI5.1
 
<tb> <SEP> organism of <SEP> test <SEP> not <SEP> taking <SEP> Penicillinē¯¯¯¯¯¯¯¯¯¯¯
<tb>
<tb>
<tb> not <SEP> the <SEP> gram <SEP> Methicillin <SEP> WG. <SEP> 832
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> Escherichia <SEP> coli <SEP> 4.3 <SEP> 5.3
<tb>
<tb>
<tb>
<tb>
<tb> Proteus <SEP> mirabillis <SEP> 4.4 <SEP> 5.5
<tb>
<tb>
<tb>
<tb>
<tb> Salmonella <SEP> schottmuelleri <SEP> 4 <SEP> 5.0
<tb>
<tb>
<tb>
<tb>
<tb> Salmonella <SEP> typhosa <SEP> 4 <SEP> 5.5
<tb>
<tb>
<tb>
<tb>
<tb> Shigella <SEP> dysenteriae <SEP> 4 <SEP> 5.7
<tb>
<tb>
<tb>
<tb> Neisseria <SEP> gororrhoeae <SEP> 6.3x <SEP> 7.2X
<tb>
<tb>
<tb> (with <SEP> susceptibility <SEP> reduced
<tb>
<tb> to <SEP> the <SEP> penicillin)
<tb>
 x Test medium:

  blood ascites substrate
By considering Table II, it appears that 3-amino-adamantyl- (1) -penicillin is on average ten times as active as methicillin with respect to the given bacteria, which should only be considered as a representative selection of various pathogenic bacteria against which the penicillins of the present invention are effective, and which serve to illustrate where this particular effect may be of importance in controlling disease. The organisms Escheria coli and Proteus .. mirabilis are, although not considered to be pathogenic in general, often involved in infections of the urinary system.



   The aforementioned advantageous properties also appear from the following tables in which 3-amino-adamantyl- (1) -penicillin is designated WG 832.



   TABLE III Therapeutic experiments against Staphylococus aureus Mice were infected intravenously with 0.2 ml of a 1/10 diluted solution of a 24 hour culture of Staphylococcus aureus producing penicillinesis, Treatment: 1 daily dose subcutaneously for 5 days starting on the day of infection.

 <Desc / Clms Page number 6>

 
 EMI6.1
 
<tb>



  Dose <SEP> Number <SEP> Animals <SEP> survived <SEP> Average life <SEP>
<tb>
<tb>
<tb> of animals <SEP> before <SEP> the <SEP> 34th <SEP> in <SEP> days
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> Witness <SEP> 22 <SEP> 0% <SEP> 3.5
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> Benzyl-
<tb>
<tb>
<tb> penicillin <SEP> 3 <SEP> mg <SEP> 20 <SEP> 20% <SEP> 10.0
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> 1 <SEP> mg <SEP> 20 <SEP> 5% <SEP> 6.0
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> 0.3 <SEP> mg <SEP> 20 <SEP> 0% <SEP> 6.6
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> Methicillin <SEP> 1 <SEP> mg <SEP> 20 <SEP> 35% <SEP> 19.3
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> 0.3 <SEP> mg <SEP> 20 '<SEP> 5% <SEP> 10.0
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> 0.1 <SEP> mg <SEP> 20 <SEP> 10% <SEP> 6,

  3
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> WG <SEP> 832 <SEP> 1 <SEP> mg <SEP> 20 <SEP> 65% <SEP> 32.4
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> 0.3 <SEP> mg- <SEP> 20 <SEP> 55% <SEP> 21.1
<tb>
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> 0.1 <SEP> mg <SEP> 20 <SEP> 15% <SEP> 7.9
<tb>
 
TABLE IV Therapeutic experiments against Staphylococcus aureus Mice with an average weight of 20 g were infected intravenously with 0.2 ml of a dilute solution 1/50 of a 24 hour culture of Staphylococcus aureus sensitive to penicillin .



  Treatment: 1 daily dose subcutaneously for 5 days starting on the day of infection.
 EMI6.2
 
<tb>



  Dose <SEP> Number <SEP> Animals <SEP> surviving <SEP> on <SEP>:
<tb>
<tb> of animals <SEP> 6th <SEP> day <SEP> the <SEP> 20th <SEP> day
<tb>
<tb>, Witness <SEP> 10 <SEP> 3 <SEP> 3
<tb>
<tb>
<tb>
<tb> Benzyl-
<tb>
<tb> penicillin <SEP> 0.1 <SEP> mg <SEP> 10 <SEP> 9 <SEP> 9
<tb>
<tb> 0.03 <SEP> mg <SEP> 10 <SEP> 10
<tb>
<tb>
<tb> 0.01 <SEP> mg <SEP> 10 <SEP> 8 <SEP> 6
<tb>
<tb>
<tb> Methicillin <SEP> 0.3 <SEP> mg <SEP> 10 <SEP> 6 <SEP> 4
<tb>
<tb>
<tb> 0.1 <SEP> mg <SEP> 10 <SEP> 6 <SEP> 5
<tb>
<tb>
<tb> 0.03 <SEP> mg <SEP> 10 <SEP> 6 <SEP> 2
<tb>
<tb>
<tb> WG <SEP> 832 <SEP> 0.3 <SEP> mg <SEP> 10 <SEP> 10 <SEP> 9
<tb>
<tb>
<tb> 0.1 <SEP> mg <SEP> 10 <SEP> 10 <SEP> 9
<tb>
<tb>
<tb> 0.03 <SEP> mg <SEP> 10 <SEP> 9 <SEP> 8
<tb>
 Considering Table III and IV, it appears!
 EMI6.3
 than 3-amino-adamantyl- (1)

  -pen3ci11W e has the same or a greater effect than the well-known methicillin against a

 <Desc / Clms Page number 7>

 infection caused by a penicillinase-producing microorganism, and that it has the much greater effect previously assumed against infections caused by microorganisms that do not produce penicillinase.



   The advantageously slow elimination from the body of the penicillins of the invention has been demonstrated both in animal experiments and in human experiments following administration of 3-amino-adamantyl- (1 ) - penicillin in the form of the ampho-ion; see the following Tables V and VI where prolonged excretion is estimated on the basis of concentrations established in the blood at suitable time intervals after the administration of 3-amino-adamantyl- (1) -penicillin, and respecrive / penicillin G and methicillin as reference compounds.



   TABLE V Serum Concentrations for Rats Receiving 20 mg (100 mg / Kg) of WG 832, Methicillin and Penicillin G-Na Sub-Cu / Year
 EMI7.1
 
<tb> The <SEP> results <SEP> are <SEP> the average <SEP> <SEP> of <SEP> those <SEP> obtained <SEP> for <SEP> 3 <SEP> rate.
<tb>
<tb>
<tb>



  Concentration <SEP> in <SEP> serum <SEP> (Ug / m1)
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> Substance <SEP> Hours <SEP> after <SEP> administration
<tb>
<tb>
<tb> 1/2 <SEP> 1 <SEP> 2 <SEP> 4 <SEP> 6
<tb>
<tb>
<tb>
<tb>
<tb> WG <SEP> 832 <SEP> 77 <SEP> 55 <SEP> 22 <SEP> 2.9 <SEP> 0.3
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> Pen. <SEP> G-Na <SEP> 27 <SEP> 3.9 <SEP> 0.35 <SEP> 0.02 <SEP> 0.03
<tb>
<tb>
<tb>
<tb>
<tb>
<tb> Methicillin <SEP> 44 <SEP> 23 <SEP> 2.5 <SEP> 0.24 <SEP> -
<tb>
<tb>
<tb>
<tb>
<tb> No <SEP> detectable <SEP> activity.
<tb>
<tb>
<tb>
 

 <Desc / Clms Page number 8>

 



   TABLE-VI Concentration in Serum for Humans Receiving 100 mg of WG 832 and Penicillin G-Na intramuscularly.



  The results are the average of those obtained on 2 subjects.
 EMI8.1
 
<tb>



  Concentration <SEP> in <SEP> serum (@ q / ml)
<tb>
<tb>
<tb> Substance <SEP> Hours ,, after <SEP> administration
<tb>
<tb> 1/2 <SEP> 1 <SEP> 2 <SEP> 3 <SEP> 4
<tb>
<tb>
<tb>
<tb>
<tb> WG <SEP> 832 <SEP> 3.8 <SEP> 3.4 <SEP> 1.6 <SEP> 1.0 <SEP> 0.41
<tb>
<tb>
<tb>
<tb>
<tb> Pen. <SEP> G-Na <SEP> 3.1 <SEP> 1.9 <SEP> 0.47 <SEP> 0.21 <SEP> 0.07
<tb>
 
Another object of the invention is to provide processes for the production of the penicillins of formula (I).



   In one embodiment of the invention, the process comprises reacting 6-amino-penicillanic acid with a functionally reactive derivative of formula (II):
 EMI8.2
 in which R1 and R have the meaning given above and COY denotes a radical capable of reacting with the amino group; with a view to forming the compound of general formula (I) and recovering the latter compound as such or in the form of one of its salts.



   It has been found, which appeared surprising, that the nitrogen atom attached to the positions of adamantane (1) -carboxylic acids substituted in position 3, applied in the process according to the invention, is so suitable for ionization that it is possible to carry out the reaction between the functionally reactive derivative of such adamant ne- (1) -carboxylic acid substituted in position 3 and 6-amino-penicillanic acid without self-condensation takes place under the formation of polymer derivatives of adamantane.,

 <Desc / Clms Page number 9>

 It has been suggested that the steric hindrance in the adamantane molecule results in suitable reactive functional derivatives of adamantane acid- (1)

  -3-substituted carboxylic acid are obtained without prior protection of the primary or secondary amino group attached to the 3-position of the adamantane molecule, since possible reaction products resulting from secondary reactions under the formation of the functionally reactive derivative of the 3-Substituted adamantane- (1) -carboxylic acid have been shown to be inferior and of importance to obtaining satisfactory yield and purity of the desired 3-substituted adamantyl- (1) -penicillin by reaction. between the functionally reactive derivative of 3-substituted adamantyl- (1) -carboxylic acid and 6-amino-penicillanic acid.



   Accordingly, it is not necessary to protect the amino group at position 3 of the adamantane molecule when carrying out the process for the production of the penicillins of the present invention. As an example, it turned out that the
3-amino-adamantyl- (1) -penicillin can be produced in good yield by simply reacting 3-amino-adamantane- (1) -carboxylic acid with, for example, thionyl chloride, and then allowing the resulting product to react with 6-amino-penicillanic acid in an aqueous medium and at a suitable pH value at which the amino group of 6-amino-penicillanic acid in all its essential characteristics is not ionized, for example at a pH value of about 5.5-7.0.



   Since the standard process for producing the penicillins of the general formula (1) in which R1 and / or R2 are hydrogen is believed to involve that the amino group attached to the adamantane molecule is protected during the reaction, for example by carbobenzoxylation, the foregoing characteristic implies a purely preparatory advantage making superfluous the final reductive cleavage of the carbobenzoxy group necessary for obtaining the desired penicillin of formula (I).

 <Desc / Clms Page number 10>

 



   Suitable functional derivatives of adamantap- (1) -carboxylic acid substituted in the 3-position include halides, such as carboxylic acid chloride, carboxylic acid anhydrides, mixed anhydrides, such as anhydride. mixed with a carbonic acid mono-ester or with a sulfonic acid, or else the imidazolide of the carboxylic acid.



   The process of the invention can be carried out following known procedures, however always taking into account the relative instability of the penicillin obtained.



   For example, it is desirable to carry out the reaction between the functional derivative of substituted admantane- (1) -carboxylic acid and 6-amino-penicillanic acid at a temperature of about 0 ° C. and preferably at a pH value close to the neutral point, for example in the presence of a suitable buffer.



  Under such conditions, the reaction will generally be complete within a period of 15 minutes to a few hours.



   The resulting penicillin can, in some cases, be isolated by methods generally used to isolate amphoteric compounds. Thus, the pH value of an aqueous solution of the compound can be adjusted to the isoelectric point of the compound in question, and the precipitated product, if any, can be collected and then purified by crystallization.



   Poorly soluble salts of the penicillins can, for example, be isolated also by a double decomposition carried out in water, or in mixtures of water and a suitable organic solvent, in a known manner.



   From its poorly soluble amino salts, penicillin can be suitably converted to its sodium salt, for example, by reaction with aqueous sodium bicarbonate in the presence of a water-immiscible solvent, so that the amine is separated. A sodium salt can then be isolated

 <Desc / Clms Page number 11>

 by freezing desiccation of the aqueous phase and it can then be subjected to further purification.



   The starting materials of formula (II) which are used in the above embodiment are heretofore unknown substances, which can be produced starting from readily available 3-bromo-adamantane- (1) -carboxylic acid. and which is described in Chemische Berichte 95,669 (1962), Stetter & Mayer.

   By treating this acid with acetonitrile in
 EMI11.1
 sulfuric acid, 3'-acetamino'-ad <nnantane- (l) - acid is obtained
Carboxylic unknown so far which, by saponification. is converted into the corresponding free amine and, if desired, the acid
 EMI11.2
 3 "amino-'adamantane- (1) - 'carboxylic acid can be mono- or dialkylated to produce 3-mono- or 3-dialkylamino <adamantane- (1) -carboxylic acids
According to another suitable embodiment of
 EMI11.3
 In the invention, the compound 3-amino-adamantyl- (1) -penicillin is prepared, starting from 3-azido-adamantane- (1) -carboxylic acid.

   hitherto unknown, in the form of one of its functionally reactive derivatives, for example the halide, which is reacted with 6-amino-penicillanic acid, 3-azido-adamantyl The resulting (1) -penicillin then being hydrogenated to form the desired 3-amino-adamantyl- (1) -penicillin.



   It has been found that it is most particularly suitable, in the latter process, to carry out the hydrogenation at a determined pH value, close to neutral, which can be obtained by successive neutralization of the strongly basic amino group formed under hydrogenation. , for example by automatic addition of acid.



   In this embodiment, 3-amino-adamantyl- (1) -penicillin is formed in yields and purity which are extremely good, since side reactions are avoided, and since hydrogenation can be made optimum by at the con-

 <Desc / Clms Page number 12>

   trOle of the process by the acid consumed in connection with the neutralization mentioned above.

   
 EMI12.1
 2-Aziclo-adamantane- (1) -carboxylic acid, used as a starting material in the latter embodiment, can be obtained from 3-bromo-adamantane- (1) -carboxylic acid , the latter compound being reacted with lithium azide in an inert and anhydrous solvent, for example dimethylformamide.
 EMI12.2
 



  3-arnino-adamantyl - (.} - pniciJ.line prepared as above has a melting point of 236 C (decomposition) and an [a] 20D value of + 2530 (in water), Sa solubility in water as an ampho-ion is about 4.5% (by weight), an elemental analysis of a product with a water content of 4.8% ( by weight), determined by the Karl Piocher method, giving the following results Found C "55.65; H: 7.15t N '10.10%
 EMI12.3
 Calculated for CxF273Q.i3C Ci 55.46? Is 7.10; Nt 10.21%
The present invention further relates to therapeutic compositions applicable in the treatment of infectious diseases.

   The composition of the invention contains, as a therapeutic component, at least one member of the class comprising the penicillins of general formula (I) and their salts with organic or inorganic bases, non-toxic, pharmaceutically acceptable, as a mixture. with auxiliary agents and solid or liquid pharmaceutical carriers.



   In this composition, the therapeutically active material can constitute 1 to 95% by weight of the composition.



   The composition in question can be converted into pharmaceutical presentation forms, for example tablets, pills, dragees or suppositories:., Or the composition can be placed in receptacles, such as capsules or ampules. , or again, in the case of mixtures

 <Desc / Clms Page number 13>

 or ointments, the composition can be placed in bottles, tubes or the like.



   To complete these compositions, solid or liquid, organic or inorganic, pharmaceutical carriers suitable for enteral, parenteral or local administration can be used. Water, gelatin, lactose, starches, magnesium stearate, talc, vegetable and animal oils and fats, benzyl alcohol, gums, polyalkylene glycols, petroleum jelly, butter cocoa, lanolin or other known carriers for drugs are all suitable as carriers, while stabilizers, wetting agents, or emulsifiers can be used as auxiliary agents,

   salts to vary the osmotic pressure or buffers to provide a pH value appropriate to the composition.



   Another object of the present invention is the selection of a suitable dose of the penicillins and penicillin salts of the invention which can be administered in such a way that the desired effect is achieved without side effects.



   It has been found that the penicillins or the salts of penicillins of the invention are suitably adminsitrated in unit dosages containing not less than 100 mg, preferably 250 to 2000 mg of the penicillin in question, this amount. being calculated for the free acid.



   By the term "unit dosage" is meant a unit dose, i.e. a single dose, which can be administered. to patients and which can be easily handled and packaged, while remaining a physically stable unit dose comprising either the active material alone or a mixture of this active material and pharmaceutical diluents or carriers, solid or liquid.

 <Desc / Clms Page number 14>

 



   If the composition is to be injected, a connected ampute, vial, or similar receptacle may be provided to contain a parenterally acceptable solution or dispersion of the active material in the aforementioned unit dosage. For enteral administration, the unit dosage may preferably consist of 250 to 500 mg of the penicillin concerned, for example in the form of tablets which should advantageously be coated with an enteric coating.



   According to a suitable embodiment, however, the penicillins of the invention are administered in the form of injectable preparations, for example an aqueous or oily dispersion of penicillin, these preparations possibly containing the penicillin in question in the form of ampho-ion or one of its non-toxic salts, or in the form of mixtures of these compounds, with a view to obtaining for example a high initial effect and a desired prolonged effect ;. For this purpose, the preparation can advantageously contain both a water soluble salt of penicillin and one of its water insoluble salts.



   Among the suitable salts of the penicillins of the invention, there may be mentioned alkali metal salts, alkaline earth metal salts, ammonium salt and non-toxic amine salts, for example trialkylemines, such as. as triethylamine, procatne, dibenzylamine, N, N-dibenzylethyenediamine and other amines used to form salts with known penicillins.



   For parenteral administration, the unit dosage according to the present invention may contain from 250 to 2000 mg, preferably 1000 mg, per dose, this amount being calculated for the free acid.



   In the form of the unit dosages mentioned above the penicillins can be administered once or more times a day at appropriate intervals, taking into account however

 <Desc / Clms Page number 15>

 always the patient's condition.



   In clinical practice, the 3-amino-ada mantyl- (1) -penicillin ampho-ion has been used with success, especially in the treatment of osteomyelitis.



   In the latter treatment, an aqueous dispersion of the penicillin amphoion is used and, administered intramuscularly in doses of 1000 mg by injection.



   In connection with the treatment, it has been established that the concentration of penicillin in the blood two hours after the injection amounts to 16 micrograms / ml, while it is 11 micrograms / ml 4 hours after the injection. injection and 1.5 microgram / ml 12 hours after injection, these figures being averages.



   A suitable high and constant concentration of penillin was obtained after 3 daily injections which were administered during the treatment period extending from a few weeks to several months in severe cases, until one had. noted recovery by clinical examination as well as by bacteriological examination.



  EXAMPLE 1
 EMI15.1
 3-Acetamino-adamantane- (1) -carboxyljque Acid A mixture of acid is refluxed for 20 hours.
 EMI15.2
 3-Bromo-adamantane- (1) -carboxylic (5.8 g), acetonitrile (45 ml) and concentrated sulfuric acid (9 ml).



   After cooling, the mixture is poured into water (250 ml) and the resulting suspension is concentrated in vacuo to remove most of the acetonitrile.



   An aqueous solution of caustic soda is added (33%) until the pH is 4/0 (approximately 24 ml).



   The precipitate is filtered off, washed with water and dried to give 4.4 g of the desired product, which has a melting point of 23-240 ° C. Two recrystallizations from metha-

 <Desc / Clms Page number 16>

 nol-acetonitrile raise the melting point to 254-258 C.
 EMI16.1
 3-Amino-adamantane- (1) -carboxylic acid
A solution of 3-acetamino-adamantane- (1) -carboxylic acid (3.0 g) in 4N caustic soda (40 ml) is subjected to reflux for 5 hours. After cooling; the pH of the solution is adjusted to 7 with acetic acid. The crystalline precipitate is filtered off, washed with ethanol and dried to give 2.20 g of the desired compound which has a melting point above 330 C.

   To purify the compound, 2.0 g of it is suspended in water (10 ml). 4N NaOH (2 ml) is added and the resulting solution is filtered through a known filter aid. under the brand "Dicalice". The filtrate is adjusted to a pH of 6.5 with acetic acid. The crystalline precipitate thus formed is separated by filtration, washed with a little water and then with alcohol, and dried to give 1.55 g of pure 3-amino-adamantan- (1) -carboxylic acid.



   Analysis: Calculated for: C11H17NO2:
C: 67.66; H: 8.78; N: 7.17%
Found: C, 67.49; H: 8.93; N: 7.24% EXAMPLE 2
 EMI16.2
 3-amino-adamantyl- 1) -penicillin
3-Amino-adamantane- (1) -carboxylic acid (1 g) is refluxed with thionyl chloride (4.6 ml) for
1 hour. The excess thionyl chloride is removed in vacuo. The residue is dissolved in benzene (3 ml) and the resulting solution is evaporated under reduced pressure to remove traces of thionyl chloride. This process is repeated to leave 1.3 g of a solid, almost colorless residue.

   Elemental analysis and infrared spectrum suggests that this product is 3-thionylimino-adamantane- (1) -carboxylic acid chloride. 500 mg of this product is dissolved in dry acetone (8 ml) and the resulting solution is added over 20 minutes with stirring.

 <Desc / Clms Page number 17>

 to a suspension of 6-amino-penicillanic acid (520 mg) in 50% aqueous acetone (20 ml), which had previously been adjusted to a pH of 7.0 with thriethylamine. During the process, a vapor of pH 7.0 p is maintained by the addition of an IN solution of triethylamine in 50% aqueous acetone from an automatic titration apparatus, at the end of the process. reaction, which is carried out at room temperature, gives a clear solution.

   The acetone is removed in vacuo and the resulting aqueous solution is tested for its antibacterial activity by the agar-cup method, using Sarcina lutea as the test organism. , and a pure sample of 3-amino-adamantyl- (1) -penicillin as a reference compound. According to this test, 70% of the theoretical amount of 3-amino-adamantyl- (1) pnicillin is formed. Paper chromatography reveals that only one antimicrobial compound is present and that this compound is identical to the authentic 3-amino-adamantyl- (1) -penicillin.



   The aqueous solution is concentrated in vacuo to a volume of 4 ml. Addition of acetone (40 ml) gives an oily precipitate which, after decomposition and addition of fresh acetone, gives 900 mg of a semi-crystalline solid which contains 50% of the theoretical amount of 3-amino-adamantyl- ( 1) -penicillin.



  EXAMPLE 3
 EMI17.1
 3-amino-adamantyl- L-penicillin
3-Amino-adamantyl- (1) -carboxylic acid (1.0 g) is refluxed with thionyl chloride (4.6 ml) for 1 hour. After cooling, the excess thionyl chloride is removed in vacuo. The residue is dissolved in dry benzene (3 ml) and the solution is evaporated in vacuo to separate the traces of thionyl chloride. The residue (1.3 g) is dissolved in dry benzene (20 ml) and dry hydrochloric gas is bubbled through the solution for 15 minutes to give a deposit of one.

 <Desc / Clms Page number 18>

 colorless crystalline material. The mixture is evaporated under reduced pressure to leave 1.3 g of 3-amino acid chloride.
 EMI18.1
 adanantane- (1) -carboxylic in the form of the hydrochloride.

   This material is added at room temperature in small portions to a stirred suspension of 6-amino-penicillanic acid (520 mg) in water (10 ml) which has previously been adjusted to a pH of 5. 5 with 1N aqueous NaOH. During this phase, a pH of 5.5 is maintained by the addition of 1N aqueous NaOH, this addition being controlled by an automatic titration apparatus.



   The resulting solution is tested for antibacterial activity by the agar-cup method using Sarcina lutea as the test organism, Following this test, about 30% of the theoretical amount of 3 -amino- adamantyl- (1) -penicillin were formed. Chromatography on paper reveals that only an antibacterial compound is present in the soluti and that this compound is identical to 3-amino-adaman-
 EMI18.2
 tyl- (1) -pënicill..ne authentic.



  EXAMPLE 4 3-Amino-adamantvl- (1) -penicillin a. A solution of 3-amino-adamantane- (1) -carboxylic acid (195 mg) in 2N caustic soda (1 ml) is cooled in ice water and carbobenzoxy chloride (0, 18 ml). The mixture is shaken thoroughly for 10 minutes. After acidification with hydrochloric acid, the mixture is extracted with ether. The ethereal phase is washed with water, dried and '' evaporated to dryness to give 200 mg of 3-carbobenzoic acid.
 EMI18.3
 xyamino-, adamantane -. (1) - carboxylic with a melting point of 112-1140C.

   The analytical sample, which is obtained after recrystallization from ether-hexene, has a melting point of 115-115.5 C. Analysis: Calculated for: c19H23NO4; C: 69.28; H: 7.04; N: 4.25%
Found: C: 69, d9; H: 7.08; N: 4.19%

 <Desc / Clms Page number 19>

 b, 3-Carbobenzoxyamino-adamantane- (1) -carboxylic acid (500 mg) is refluxed with thionyl chloride (4 ml) for 90 minutes. The excess thionyl chloride was removed in vacuo and the residue, which was crude 3-carbobenzoxyamino-adamantane- (1) -carboxylic acid chloride, was dissolved in acetone (7 ml).

   The resulting solution is added at 0 C to a solution of 6-amino-penicillanic acid (330 mg) in a 3% solution of sodium bicarbonate (10 ml) with stirring,
After stirring for 30 minutes, the mixture is extracted with ether. The aqueous phase is separated, adjusted to pH 2.5 with dilute sulfuric acid and then immediately extracted with ether. The ethereal extract is washed with water and
 EMI19.1
 dried, after which benxyl-p-phnylethylamine (0.2 ml) is added to precipitate the crystalline benz1-phenylethylamine salt of 3-carbobenzoxyamino-adamantyl- (1 () penieillin, which is collected, washed. with ether and dried to give a product of 89% purity c.

   Suspended in sodium bicarbonate
 EMI19.2
 3% aqueous (15 ml), 650 mg of the 3-carbobenzoxyamino-adamantyl- (1) -pdnicillin ben2yl-phenylethylamine salt as described above. Ethyl acetate (15 ml) is added and the mixture is stirred until two clear phases are obtained. The aqueous phase is separated and added to a prehydrogenated suspension of 5% palladium on calcium carbonate (5 mg) in water (15 ml), after which the mixture is shaken under a pressure of an atmosphere of. hydrogen.

   When the hydrogen consumption ceases (after 30 minutes), the catalyst is filtered off and washed with water, The combined filtrate and washings are adjusted to pH 6 with dilute sulfuric acid and dried. to give 400 mg of 3-amino-adamantyl- (1) -penicillin.

 <Desc / Clms Page number 20>

 



  EXAMPLE 5 3-Amino-adamantvl- (1) -penicillin a. A mixture of 3-bromo-adamantane- (1) -carboxylic acid (5.0 g) and lithium azotbhydride (10 g) in dimethylformamide (40 ml) is refluxed for 18 hours. . After cooling, the solution is concentrated in vacuo and the residue is dissolved in water. The solution (40 ml) is acidified with hydrochloric acid and extracted with ether (2 x 25 ml).



  The combined extracts are washed with water, dried and evaporated to give 2.5 g of a product which, in addition to the 3-azido-adamantyl- (1) -carboxylic acid compound, contains a certain amount of acid.
 EMI20.1
 3-hyeroxy-adamantane = (1) -cai #> oxyliqyxT This latter compound is insoluble in carbon tetrachloride and is separated by treatment with this solvent. This filtrate, which contains 3-azido-adamantyl- (1) -carboxylic acid, is evaporated to dryness and the residue is chromatographed on a 50 g column of silica gel. Elution with pentane-ether (85/15) gives 1.3 g of pure 3-azido-adamantyl- (1) -carboxylic acid, melting point 101-102 C.



   The infrared spectrum (KBr) shows a strong band at 2110 cm-1b. 3-Azido-adamantyl- (1) -carboxylic acid (1.55 g) is refluxed with thionyl chloride (3 ml) for 1 hour. The excess thionyl chloride is removed in vacuo and the residue, which is crude 3-azido-adamantyl- (1) -carboxylic acid chloride, is dissolved in acetone (25 ml). The resulting solution is added over a period of 15 minutes at 5 ° C to a solution of 6-amino-penicillanic acid (1.53 g) in 3% sodium bicarbonate solution (30 ml) with stirring.



  After stirring for 1 hour at 5 ° C., the mixture is washed twice with 25 ml of ether. The aqueous phase is covered with

 <Desc / Clms Page number 21>

 : layer of 50 ml of ether and it is adjusted to a pH of 2.1 with dilute sulfuric acid, the free penicillin passing into the organic phase. The layers are separated and the aqueous phase is extracted twice again with 5 ml portions of ether, water (1 ml) is added to the combined ethereal lines and then a concentrated aqueous solution of bicar is added. - potassium bonate with stirring until the pH of the aqueous phase is 7.0.

   The aqueous phase is separated, n-butanol (50 ml; and the mixture is distilled in vacuo to separate the water, additional amounts of n-butanol are added and the distillation is continued until all l. water is separated.



  During this process, the potassium salt of 3-azido-adamantyl- (1) - penicillin crystallizes. It is filtered off, washed with acetone and dried to give 2.1 g of a product of 95% purity. vs. A solution of the potassium salt of 3-azido-adamantyl- (1) -penicillin (400 mg) in water (10 ml) is added to a prehydrogenated suspension of 5% palladium on calcium carbonate ( 800 mg) in 5 ml of water. The mixture is shaken under a pressure of an atmosphere of hydrogen for 1 hour. The catalyst is filtered off and washed with water (2 x 10 ml). The combined filtrate and washings are adjusted to pH 6 with dilute hydrochloric acid and dried to give 330 mg of 3-amino-adamantyl- (1) -penicillin having a purity of 72%.



   The infrared spectrum shows that the strong azido band has 2110 cm-1 in the spectrum of the starting material disappeared, while the 1775 cm-1 band due to the O-lactam nucleus is still present.



   A paper chromatograph (solvent water / ethyl acetate Nature!) "Whatman" paper n 1 buffered with 10% sodium citrate at a pH of 5.6) is placed on an agar plate seeded with staphylococcus aureus. After incubation at 37 ° C, a single well-defined zone of inhibition is observed near the

 <Desc / Clms Page number 22>

 starting place, while no other zone of inhibition can be detached ("Whatman" is a registered trademark).



  EXAMPLE
 EMI22.1
 3 m3.no-.adamant 1-. 1 Sn.a3. not
A 10% prehydrogenated palladium suspension of clacium carbonate (12 g) in water (250 ml) is adjusted to a pH value of 8.5. A solution of 22.8 gr (50 mmol) of the potassium salt of 3-azido-adamantyl- (1) -penicillin in 100 ml of water is added slowly with stirring, while a stream of hydrogen is bubbled through. through the suspension. During the process, a pH value of 8.5 is maintained by addition of 2N acetic acid, the addition being monitored by an automatic titration apparatus.

   When the hydrogenation is complete (i.e. when the acid consumption has ceased), the catalyst is removed by filtration and washed on the filter with 3 portions of
25 ml of water, the combined filtrates and washings then being treated to pH 6.50 with acetic acid. Normal butanol (750 mL) was added and the resulting mixture was concentrated in vacuo, the water being azeotroped off. the residual butanol solution, water (3 ml) is added and crystallization is initiated by scraping or seeding. After standing overnight in a refrigerator, crystals which form a hydrated form of the desired product are collected, washed with normal butanol, then with acetone, and finally they are dried.

   The product thus obtained is recrystallized in the following way from a solution of 10 g (100 ml), acetone (600 ml) is added slowly with stirring. After standing, the crystals are separated by filtration, washed with acetone and dried to give 8.0 g of the desired penicillin in the form of its monhydrate. The melting point is 235 c (decomposition) and the optical rotation [a] 20 is +253 (in water).

 <Desc / Clms Page number 23>

 



  Analysis: Calculated for C19H27N3O4S.H2O
C: 55.46; H: 7 ', 10; N: '10.21%
Found: C: 55, 67; H. 7.15; N: 10.10%
The water content is determined by the karl Fischer method to be 4.8%. Pure penicillin has a water solubility of about 4.5% as the ampho-ion.



  EXAMPLE
 EMI23.1
 3-dimthylamino-adamaltyl- (1) - tenicillin a. 3dimdthy.amino-adamantane- (1) -carboxxlic acid
 EMI23.2
 To a solution of 3 amino-adamantanct (1) -caboxylic acid (10 gr) in an aqueous solution of 0.5 M caustic potassium hydroxide (100 ml) is added a 35% aqueous solution of formaldehyde (8 ml). and, after standing for 15 mls, a catalyst containing 10% palladium on calcium carbonate (6 g) is added.



   The mixture is shaken under an atmosphere of hydrogen at about 50 ° C. For 24 hours, 2.3 liters of hydrogen are consumed, then consumption ceases.



   After separation of the catalyst by filtration, the filtrate is adjusted to pH 7.0 with dilute hydrochloric acid, then it is evaporated to dryness. The residue is treated with 250 ml of hot ethanol and the mixture is separated. by filtration the insoluble material (KCl) and the filtrate evaporated in vacuo. The crystalline residue (12 g) is recrystallized from ethanol to give 8.8 g of the desired compound.



  Analysis Calculated for C13H21No2 1/2 H20
VS; 67.31: H: 9.54; Ni 6.04%
Found: c, 67.70; H: 9.46; Ni * 5.98% b, 3-dimethylamino-adamantyl- (1) -penicillin
3-Dimethylamino-adamantane- (1) -carboxylic acid (1.6 g) is refluxed with thionyl chloride (3 ml) for 1 hour, the excess thionyl is removed in vacuo and the residue which consists of 3-dimethylamino-ada- acid chloride

 <Desc / Clms Page number 24>

 
 EMI24.1
 crude mantan- (1) -carboxylic as the hydrochloride, ¯ is suspended in dry acetone (30 ml) and the suspension is added over a period of 20 minutes with stirring to an acid solution 6 -amino-penicillanic acid (1.53 gr) in a 3% aqueous solution of potassium bicarbonate- (50 ml).

   After stirring for 1 hour at 0-5 c, the mixture is washed twice with 50 ml of ether. The aqueous phase is adjusted to a pH value of 6.5 with 2-ethylhexanoic acid and concentrated in vacuo to a volume of 20 ml.



  Addition of acetone (80 ml) gives the desired penicillin as a white amorphous powder having a purity of 85%.



   By substituting the acid 3-methyl-ethylamino-adamantane-
 EMI24.2
 (l) -carboxylic and 3-diethylamïno-adamantane - (1) -carboxylic acid with 3-dimethylamino-adamantane- (1) -carboxylic acid in the preceding process, 3-'methyl is obtained -ethylamino-adamantane- (1) -penicillin. and 3-diethylamino-adamantane- (1) -penicillin.



  EXAMPLE 8
 EMI24.3
 3-Amino-adamantyl- (1) -penicillin triethylamine salt To a suspension of 3-amino-adamantyl- (1) -penicillin (410 mr) in water (3 ml) is added triethylamine ( 0.15 ml). Addition of acetone (15 mL) to the resulting clear solution causes the desired compound to crystallize as colorless crystals which are collected, washed with acetone and dried in vacuo.
In the same way, the salts with tri-methylamine, methyldiethylamine, triethanolamine and ethyl-piperidine are prepared.



  EXAMPLE 9
 EMI24.4
 Sodium salt of 3-amino = adamantyl- (11-nenicillin To a suspension of 3-amino-adamantyl- (1) -penicillin (410 gr) in water (3 ml) is added an aqueous solution of soda caustic lN (1.0 ml), and this dropwise with stirring,

 <Desc / Clms Page number 25>

 Addition of acetone (20 ml) precipitates the desired sodium salt as an amorphous powder which is separated by filtration, washed with acetone and dried in vacuo at room temperature.



   In a similar fashion, the salts of lithium, potassium and clacium are prepared.



   CLAIMS 1, Penicillin derivatives of the general formula
 EMI25.1
 wherein each of R1 and R2 is a hydrogen atom or a lower alkyl radical containing from 1 to 4 carbon atoms, and the readily hydrolyzable esters or the pharmaceutically acceptable salts of these penicillins.



   2. 3-Amino-adamantyl- (1) -penicillin and its readily hydrolyzable esters or pharmaceutically acceptable salts.



   3. 3-Dimethylamino-adamantyl- (1) -penicillin and its readily hydrolyzable esters or pharmaceutically acceptable salts.
 EMI25.2
 



  4. 3-Methyl ethylamino-adamantyl- (1) -penicillin and its readily hydrolyzable esters or pharmaceutically acceptable salts.

** ATTENTION ** end of DESC field can contain start of CLMS **.

 

Claims (1)

5. The sodium, potassium, magnesium, ammonium and calcium salts of 3-amino-adamantyl- (1) -penicillin, EMI25.3 6. Salts of 3-amino-adamantyl- (1j-penicillin with pharmaceutically acceptable organic bases. <Desc / Clms Page number 26> 7. The crystalline monohydrate of the 3-amino-adamantyl- (1) -penicillin ampho-ion. 8. A process for the production of penicillins according to claim 1 which comprises reacting 6-amino-penicillanic acid with a functionally reactive derivative of the formula. EMI26.1 in which R1 and R2 have the meaning given above and COY denotes a radical capable of reacting with the amino group to ensure the formation of the compound of general formula (I), and the recovery of the latter compound as it is or in the form one of its salts. 9. A process according to claim 7, wherein: the compound of general formula (II) is a carboxylic acid halide. EMI26.2 10. A process for the production of 3-amino-adamantyl- (I) - penicillin, which comprises hydrogenating a member of the class comprising Ia. 3-azido-adamantyl- (l) -penicillin and its salts, and EMI26.3 recovering the 3-amino-adamantyl- (1) -penicillin formed as such in the form of one of its salts. 11. As an intermediate, 3-acetamino-adamantane- (1) -carboxylic acid. 12. As intermediates, 3-amino-adamantal- (1) -carboxylic acid and its halides, and their halohydrates. 13. As intermediates, 3-carbobenzoxy-amino-adamantane- (1) -carboxylic acid and its halides. 14. As intermediates, 3-carbobenzoxyamino-adamantyl- (1) -penicillin and its salts. 15. As intermediates, 3-azido-adamantyl- (1) -carboxylic acid and its halides. <Desc / Clms Page number 27> As intermediates, 3-azido-adamantyl- (1) -penicillin and its salts. 17. A therapeutic composition for use in the treatment of infectious diseases comprising, as a therapeutically active compound, a member of the class comprising the penicillins of claim 1 and their pharmaceutically acceptable salts and readily hydrolyzed esters. bles, in admixture with a material of the class comprising pharmaceutical carriers and auxiliary agents, solid and liquid, the proportion of the therapeutically active material of said class being between 1 and 95% of the total composition. 18. For the treatment of infectious diseases, a readily acceptable unit dosage containing at least one member selected from the groups comprising the penicillins of claim 1 and their readily hydrolyzable esters and pharmaceutically acceptable salts, the unit dosage having a content of. this active ingredient by antibiotic route from 100 to 2000 mg in total. 19. A unit dosage according to claim 18, wherein the relevant group member is admixed with a vehicle. 20. An assay according to claims 18 or 19, wherein the member of the group involved is 3-amino-adamantyl- (1) -penicillin, the dosage unit having a content of this penicillin which is from 250 to. 1000 mg, preferably in the form of its ampho-ion. 21. Penicillins and their derivatives, as described above, in particular in the examples given. 22, The production of penicillins, as described above, in particular in the examples given.