BE879925A - NEW PROCESS FOR THE PREPARATION OF A SELECTIVELY PROTECTED N-ACYL DERIVATIVE OF AN AMINO-GLYCOSIDE ANTIBIOTIC - Google Patents

Description

       

  "New process for the preparation of an N-acyl6 derivative selectively protected from an aminoglycoside antibiotic" The present invention relates to a new process

  
  <EMI ID = 1.1>

  
ment protected or blocked by an acyl group. It therefore relates to a novel process for the selective protection of amino or nlkylamino groups which occupy particular positions in the aminoglycoside antibiotic. Its main application lies in the preparation of an N-ocylated derivative selectively protected from the antibiotic

  
  <EMI ID = 2.1>

  
tion is applicable can be defined more specifically as an aminoglycoside antibiotic

  
  <EMI ID = 3.1>

  
alkanoyl) -aminoglycosides, which are known as useful semi-synthetic antibacterial agents, active against resistant bacilli.

  
Aminoglycoside antibiotics such as ka-

  
  <EMI ID = 4.1>

  
amino and several hydroxyl functions having relatively high and diverse degrees of reactivity. Many kinds of aminoglycosidic antibiotics have been synthesized

  
  <EMI ID = 5.1>

  
is often necessary or preferable to cause certain amino groups and / or certain hydroxy groups to

  
  <EMI ID = 6.1> <EMI ID = 7.1>

  
and they do not pose a problem as far as

  
  <EMI ID = 8.1>

  
on the other hand, for the selective protection of certain particular amino groups in the large number of those which comprise the aminoglycosidic antibiotic, the methods which are currently available are difficult to implement.

  
  <EMI ID = 9.1>

  
In fact, all of the amino groups of the aminoglycoside antibiotic show little difference between them.

  
  <EMI ID = 10.1>

  
tion of this problem is provided by the 6'-amino group of kanamycin A. This amino or methylamino group, which is linked to a certain carbon atom itself linked to a single carbon atom in the aminoglycosidc molecule

  
  <EMI ID = 11.1>

  
same molecule. For this reason, the first amino or methylamino group mentioned can react much more preferentially with an acylating agent, comprising a

  
  <EMI ID = 12.1>

  
  <EMI ID = 13.1>

  
deny cited, whence it follows that the N-protected derivative in which the first cited group is preferentially blocked by the acyl group can be obtained with a yield much higher than that which is possible for N- derivatives protected in other positions. There is

  
several years ago, several of the authors of the present invention have found that when an amino group and a hydroxyl group are neighbors in a pair in the steric configuration of the molecule of the antibiotic aminoglycoside, these two groups, amino and hydroxyl, little - <EMI ID = 14.1>

  
chosen from the group comprising copper (II), nickel (II), cobalt (II) and cadmium (II) is reacted, in an inert organic solvent, with an antibiotic

  
  <EMI ID = 15.1>

  
6-0 (aminoglycosyl) -2-deoxystreptamines, represented by kanamycins, gentamycins and sisomycin, this bivalent transition metal cation is complexed with a pair of groups, one amino, the other hydroxyl, which exist in particular positions of "vicinal" relationship in the aminoglycoside molecule, thus forming

  
  <EMI ID = 16.1>

  
transitional tal (cf. Japanese patent application in 1st publication under n [deg.] Sho-52-153944 and U.S. patent application N [deg.] 697 297, for which a patent was

  
  <EMI ID = 17.1>

  
this aminoglycoside antibiotic-transition metal cation complex, the complexed amino group is blocked by the cation of the bivalent transition metal. When this complex is subsequently reacted with an acylating reagent having an acyl group, only the uncomplexed amino groups

  
of the complex, which are not blocked by the bivalent metal cation, can be largely acylated, so that selective N-protection by the acyl group is obtained. This is illustrated below by taking kanamycin A as an example.

  
When reacting with kanamycin A a bivalent transition metal cation (11 ++) chosen from the group comprising copper (II), nickel (II), cobalt (II) and cadmium ( II), the complexing reaction of the bivalent metal cation (M ++) occurs between the <EMI ID = 18.1>

  

  <EMI ID = 19.1>


  
In the above complexing reaction, it can therefore be seen that at least 2.moles of the transition metal salt are required for 1 mole of kanamycin A. Lea groups 1amino and 3-amino are blocked at the same time in the metal complex resulting. When the complex of formula (I) is treated with an acylation reagent having an acyl group available as an amino-protecting group known in the traditional synthesis of polypeptides,

  
  <EMI ID = 20.1>

  
5253-5254 (197817.

  
The authors of the present invention have recognized the fact which has just been reported, but they have carried out <EMI ID = 21.1>

  
semi-synthetic derivatives of aminoglycoside antibiotics. As a result of this research, they found that although the bivalent zinc cation behaves markedly different from those of the bivalent cations of the metals listed above, nickel, cobalt, copper and cadmium, it is ultimately capable of complexing

  
  <EMI ID = 22.1>

  
3uamino group (or 3 "-alkylamino) and block these groups in an aminoglycoside compound (such as kanamycin A, B or C) which has a deoxystreptamine moiety

  
  <EMI ID = 23.1>

  
noglycosyl is linked to the 6-hydroxyl group which it contains.

  
According to Nagabhushan et al., It can be expected that when a cation of nickel, cobalt, copper or cadmium

  
  <EMI ID = 24.1>

  
ple, a complex of kanamycin B and metal salt is formed having the formula (II) below:

  

  <EMI ID = 25.1>
 

  
  <EMI ID = 26.1>

  
pairs of vicinal amino and hydroxyl groups would form reversible complexes with bivalent transition metal cations, taking into account that kanamycin B contains three of these pairs, namely between positions 1- and 2 "-, between positions 2 '- and 3'- and between positions 2 "- and 3" - of the kanamycino B molecule.

  
  <EMI ID = 27.1>

  
found that when kanamycin B is reacted with a bivalent metal cation, the zinc cation, the kanamycin B and zinc salt complex that actually forms contains the 2'-amino group and the 3 'group -Free hydroxyl, not blocked by the zinc cation. Even if the complexing reaction of the zinc cation with the 2'-amino and 3-hydroxyl groups occurs, the complexing force is extremely reduced, so that in practice the 2'-amino and 3'-hydroxyl groups are not substantially not blocked. This is why, when the kanamycin B-cation zinc complex is then acylated by reacting it,

  
  <EMI ID = 28.1>

  
introduce the benzyloxycarbonyl group as an amino-protective acyl group, a tri-3,2 ', 6'- derivative occurs

  
  <EMI ID = 29.1>

  
have been acylated, with a higher yield than the other N-acylated derivatives, but we cannot then obtain

  
  <EMI ID = 30.1>

  
present brief). This experimental fact suggests that the zinc cation has a behavior different from that of the cations of the four aforementioned metals and of behavior differs in particular in that the zinc cation does not form

  
of complex with the pair of vicinal groups 2'-amino and 3 '-hydroxyl.

  
As another example, when kanamycin A is reacted with the zinc cation and then acylated with the benzyloxycarbonyl group (cf. formula (I) below) <EMI ID = 31.1>

  
mycino A is formed as the main encylation product when the proportion of zinc cation is expected to be slightly more than 1 mole per mole of kanamycin A. In this case, it should be noted that this acylntion reaction gives takes place simultaneously, to some extent,

  
  <EMI ID = 32.1>

  
bonyle of kanamycin A and that of initial kanamycin, not acylated, but that it does not actually cause

  
  <EMI ID = 33.1>

  
cation provided by Nagabhushan et al. of the reaction mechanism, that we should expect that the

  
  <EMI ID = 34.1>

  
Example 7 of this memo). In the specification

  
  <EMI ID = 35.1>

  
necessary to use a salt of a bivalent transition metal such as copper (I), nickel (II), cobalt (II), etc., in a total proportion of at least 2 moles per

  
  <EMI ID = 36.1>

  
kanamycinc A and transition metal salt plex, as shown in formula (I) above. Experiences

  
  <EMI ID = 37.1>

  
Unlike the cations of the above four transition metals, the zinc cation is capable of blocking the 1-amino and 3 "-amino groups of kanamycin A when used in a total proportion of at least 1 mole

  
per mole of kanamycin A. Their tests have shown that when a nickel salt is reacted in a

  
  <EMI ID = 38.1>

  
namycin A, then acylate with the bcnzyloxycarbonyl group the kanamycino A-nickel salt complex which re-

  
  <EMI ID = 39.1>

  
cine A with only a very low yield, whereas a considerable yield of this derivative would be obtained in acy- <EMI ID = 40.1>

  
the zinc aminoglycoside-cation complex has its own stability which is different from that of the amino-

  
  <EMI ID = 41.1>

  
copper (II) or cadmium (II). To complex the cation of zinc with the aminoglycoside antibiotic, we

  
  <EMI ID = 42.1>

  
which is advantageously of low cost and which presents little risk of pollution to the environment.

  
Consequently, the authors of the present invention

  
  <EMI ID = 43.1>

  
react in an inert organic solvent with an aminoglycoside antibiotic containing a deoxystreptaminc fraction in which the 6-hydroxyl group is linked

  
  <EMI ID = 44.1>

  
linked to an aminoglycosyl group, the zinc cation is complexed with amino-hydroxyl pairs which it blocks and whose position can vary according to the nature of the aminoglycosidic antibiotic, and which when reacted

  
  <EMI ID = 45.1>

  
thus formed with an acylation reagent having an acyl group which is traditionally used to introduce an amino-protecting group during the synthesis of polypeptides, this acyl reagent at least one of the groups mino of the antibiotic which no are not complexed with the zinc cation and therefore not blocked by this cation, so that the amino group thus acylated is protected, and finally only when the product resulting from acylation (c '

  
  <EMI ID = 46.1>

  
zinc cation having at least one acylated amino group) is treated with a suitable reagent which eliminates said pro- <EMI ID = 47.1>

  
In a first aspect, the invention therefore provides a process for the preparation of an N-protected derivative, selectively acylated, of an aminoglycoside antibiotic, the so-called

  
  <EMI ID = 48.1> which was formed by reacting said aminoglycoside antibiotic with a zinc salt in an inert organic solvent to produce a complex of zinc cations with

  
the N-acylated derivative selectively protected from the aminoglycoside antibiotic having the initially uncomplexed amino groups acylated, and <EMI ID = 49.1> N-acylated derivative of the aminoglycoside antibiotic with a reagent which removes zinc cations from

  
  <EMI ID = 50.1>

  
ment of the aminoglycosidic antibiotic.

  
The method according to this first aspect of the invention

  
  <EMI ID = 51.1>

  
acylated with an aminoglycoside antibiotic by acylation

  
of certain amino groups other than groups 1- and 3 "-

  
  <EMI ID = 52.1>

  
this selectively acylated N-protected derivative is useful in the chemical synthesis of 1-N-aminoacylated derivatives of aminoglycosidic antibiotics such as knnamycins, including

  
  <EMI ID = 53.1> <EMI ID = 54.1>

  
antibacterial vis - &#65533; - resistant bacilli against which aminoglycoside antibiotics paronts do not act, and their antibacterial activity against a wider variety of bacilli strains is also improved compared to said parent antibiotics.

  
The implementation of the method according to this first as-

  
  <EMI ID = 55.1>

  
The aminoglycoside antibiotic that should be used

  
  <EMI ID = 56.1>

  
of zinc (which could also be called zinc complex salt) according to the invention can be any antibiotic

  
  <EMI ID = 57.1>

  
be occasionally substituted with an aminoglycosyl group. More particularly, the aminoglycoside antibiotic available in the present invention

  
  <EMI ID = 58.1>

  
be defined as an antibiotic consisting of

  
  <EMI ID = 59.1>

  
ques of the category which one can have in the process according to the invention, one can quote the antibiotics of <EMI ID = 60.1>

  
ne C, gentamycins A, B and C, verdamycin, sisomycin and netilmycin (i.e. 1-N-ethylsisomycino), as well as the other known aminoglycosides. It goes without saying that the method according to this first aspect of the invention is applicable not only to a new aminoglycoside antibiotic, which is not yet known at present and will be discovered later, but also to

  
  <EMI ID = 61.1>

  
glycosides, which will be produced in the future by chemical transformation of known aminoglycoside antibiotics.

  
As typical examples of the aminoglycoside antibiotics to which the present invention is applicable, mention may be made of kanamycin A, kanamycin 0, kanamycin C and the deoxy derivatives of these kanamycins, as well as their 6'-N-alkyl derivatives, all represented by general formula (III) below:

  

  <EMI ID = 62.1>


  
in which R is a hydroxyl group or a group a -

  
  <EMI ID = 63.1>

  
hydroxyl group, and R4 is a hydroxyl group, an amino group or an alkylamino group in which the group

  
  <EMI ID = 64.1>

  
zinc dic-cations according to the invention by reacting the aminoglycosidic antibiotic with zinc cations, it is possible to dissolve or suspend in a suitable organic solvent, mixed or not with water, a particular aminoglycosidic antibiotic aoua the form of its free base or that of an acid addition salt of said antibiotic and add to the solution or suspension thus obtained an appropriate zinc salt in a proportion of at least 1 mole per mole of the antibiotic aminoglycoside used. Any ordinary organic solvent can be used for this purpose, as long as the zinc complex formed after the addition of <EMI ID = 65.1>

  
polar organic and in particular a greater volume of water, since the presence of the polar organic solvent and of water is capable of reducing the stability of the antibiotic aminoglycoside-zinc cation complex, which does not allow the reaction d further acylation for the introduction of the amino protecting group to give satisfactory results.

  
It is therefore desirable, for the solvent in which the zinc complex must be formed, to choose a strong organic solvent, such as dimethyl sulfoxide, but it is also possible to use aqueous dimethyl sulfoxide, dimethylformamide, aqueous dimethylformamide, a mixture of diina thylaul f oxide and dimethylformamide, tetrahydrofuran, aqueous tetrahydrofuran and even a lower alkanol such as methanol, ethanol and aqueous methanol.

  
The zinc cation can be introduced in the form of a zinc salt into the reaction system where the complex is formed. Any zinc salt formed by the reaction of a zinc cation with ordinary inorganic or organic acid can be used for the effects of the present invention. However, it is generally desirable to use a zinc salt of a weak acid, such as zinc acetate, since it is common that among metal complexes containing an amino group, a non-quaternary amino group complex with a metal salt is more stable than a complex of an ammonium type amine with a metal salt and because the use of the salt

  
  <EMI ID = 66.1>

  
formation of the relatively unstable metal complex which contains an ammonium type mine. When the zinc salt of a strong acid is used, for example

  
  <EMI ID = 67.1>

  
alkaline, like 1 * sodium acetate, in addition to zinc salt <EMI ID = 68.1>

  
Offset aminoglycoside is used in the form of its addition salt with a strong acid, such as hydrochloric acid. In this case, however, care must be taken to

  
  <EMI ID = 69.1>

  
neutralizing agent, otherwise precipitation of zinc hydroxide may occur which will disturb the formation

  
of the complex. For example, when using for the formation of the complex a tetrachlorhydrate of the aminoglycoside antibiotic, it is preferable to add 4 moles of sodium hydroxide to neutralize the reaction mixture.

  
The complexing reaction can continue as long as the total molar amount of the zinc salt

  
which is used is at least equal to the total molar amount of the aminoglycoside antibiotic. It is however preferable to use the zinc salt in a proportion substantially greater than 1 mole per mole of the aminoglycosidic antibiotic, so that the balance of

  
the complexing reaction is shifted in favor of the formation of the complex. An attractive zinc complex yield can be obtained by using the zinc salt in an amount of 2.3 to 6 moles per mole of the aminoglycoside, but it is much preferable in practice to use the zinc salt in a proportion between

  
4 and 5 moles per mole of aminoglycoside. The time required for a complete complexing reaction after the addition

  
  <EMI ID = 70.1>

  
organic vant used and it can be between "instantly" (when using an aqueous organic solvent) and 20 hours. The complexing reaction can be carried out

  
  <EMI ID = 71.1>

  
also reheat or cool.

  
In this way a solution or suapen-

  
  <EMI ID = 72.1> <EMI ID = 73.1>

  
lon the invention may be a common amino protective reagent, and it is used to cause the nmino free, uncomplexed groups in the aminoglycoside antibiotic complex-zinc cations obtained to be acylated and blocked by the acyl group of the reagent cited above. The acyl group can be an alkanoyl group, a group

  
  <EMI ID = 74.1>

  
sulfonyl, all of which are conventional amino protecting groups. The available aylating reagent &#65533; this effect can be either a carboxylic acid having the following general formula (IVa):

  

  <EMI ID = 75.1>


  
in which R5 is hydrogen, an alkyl group,

  
  <EMI ID = 76.1>

  
of carbon, an aryl group, in particular phenyl, or an aralkyl group, especially benzyl, cos groups being occasionally still substituted, or an acid halide, an acid anhydride or an active ester of said carboxylic acid (IVa ), or a chloroformate of general formula (IVb):

  

  <EMI ID = 77.1>


  
  <EMI ID = 78.1>

  

  <EMI ID = 79.1>


  
either an active ester of N-hydroxysuccinimide having the following general formula:

  

  <EMI ID = 80.1>


  
or again an azidoformiatc of formula (IVc):

  

  <EMI ID = 81.1>


  
  <EMI ID = 82.1>

  
finishes that in formula (IVa), finally an acid <EMI ID = 83.1>

  

  <EMI ID = 84.1>


  
  <EMI ID = 85.1>

  
core substitutes, or an acid halide, an acid anhydride or an active ester of said sulfonic acid. It is evident, therefore, that the acylation reaction for

  
  <EMI ID = 86.1>

  
  <EMI ID = 87.1>

  
tyle, propionic anhydride, ester of trifluoroacetic acid and p-nitrophenol, ester of trifluoroacetic acid-

  
  <EMI ID = 88.1>

  
carbonyl, tosyl chloride, mesyl chloride and others.

  
The acylation reagent can be added, either as

  
  <EMI ID = 89.1>

  
aminoglycoside-zinc antibiotic complex. The molar amount of added acylating reagent can usually be equal to or slightly greater than the number of non-complex amino groups with which the acylating product is to react. In some cases, however, the molar amount

  
  <EMI ID = 90.1> the number of uncomplexed amino groups. The acylation reagent can be added, either all at once or

  
  <EMI ID = 91.1>

  
hours, however it can usually be added to

  
  <EMI ID = 92.1>

  
acylation can be carried out at a temperature included

  
  <EMI ID = 93.1>

  
kill at a temperature between 0 * C and room temperature. In some cases the reaction temperature can be kept low at the time of addition

  
  <EMI ID = 94.1>

  
as acylation progresses. Normally, the acylation reaction may not take place in situ in the organic solvent in which the aminoglycoid antibiotic-zinc catfona complex has been formed. This acylation of the zinc complex produces the N-acylated zinc complex, it

  
  <EMI ID = 95.1>

  
selectively N-acylated from the aminoglycoside antibiotic.

  
In the process according to the first aspect of the invention, the step of acylation of the aminoglycoside antibiotic complex and zinc cations is followed by that of the elimination of the latter from the Nacylated complex (in particular the destruction of the zinc complex) to give the selectively protected N-acylated derivative of the aminoglycoside antibiotic, which is released from its zinc cations.

  
To remove the zinc cations from the N-acylated zinc complex, it is necessary to treat the said complex with an appropriate reagent which removes the cations. For this, we have a large number of methods. The first is to react a zinc precipitation agent, which is capable of converting zinc cations to a water-insoluble zinc compound, such as zinc sulfide, zinc hydroxide or zinc carbonate, while the N-acylated zinc complex always remains dissolved in the acylation reaction mixture, where the aminoglycoside-zinc cation antibiotic complex has been acylated, or after it has been transferred from said re-mixture

  
  <EMI ID = 96.1>

  
tout organic fresh.

  
Among the zinc precipitation agents available for the implementation of this first method, mention may be made of hydrogen sulfide, an alkali metal sulfide, such as sodium sulfide, ammonium sulfide, a metal sulfide. alkaline earth, such as calcium sulfide, an alkali metal carbonate, such as sodium carbonate, or ammonium hydroxide. In some cases, the removal of zinc cations from the N-acylated zinc complex can be accomplished by simply adding water. In this first method, the addition of the zinc precipitation agent to the solution of the Nacylated zinc complex causes comparatively rapid precipitation of the insoluble zinc compound formed with the zinc cations, and the precipitate can be removed by filtration.

   The N-acylated derivative of aminoglycosidic antibiotic which then remains in the filtered solution can be recovered by concentration of the solution or by extraction out of this solution and, if necessary, it can be purified later. For this purification, recourse may be had, for example, to chromatography on a column of silica gel.

   A second method consists in (a) concentrating, dry or not, by evaporation of the solvent or (b) diluting with a liquid diluent, the abovementioned acylation reaction mixture or the new solution obtained by transferring the zinc N complex -acylated in a volume of fresh organic solvent, so as to obtain a deposit, concentrated or oily or solid residue, after which the desired N-acylated derivative of the aminoglycoside antibiotic, derivative present in the deposit, is recovered in any way , the concentrate or the residue. The liquid diluent which can be used in this second method is water or an organic liquid in which the solubility of the N-acylated zinc complex taken as a whole or that of its fraction constituted by the N-acylated derivative of antibiotic that aminoglycoaidic is weak or zero.

  
According to this second method, the mixture of the acylation reaction containing the N-acylated zinc complex (or the new solution obtained by transferring the said complex into fresh organic solvent) is concentrated firstly, dry or not, to obtain the oily or solid deposit or reaid. When a hardly evaporable organic solvent, such as dimethyl sulfoxide, etc., is used as the reaction medium for the N-acylation of the zinc complex, it is possible to incorporate the acylation reaction mixture containing the zinc complex Nacylated a diluting organic liquid such as ordinary ether so that the said difficult-to-evaporate organic solvent chosen as medium dissolves in the diluent or is diluted by it, which causes the deposition of a solid or oily substance containing the complex N-acylated zinc.

   An oily or solid deposit or residue is obtained in these various ways, which is normally a compound mixture: (a) of the N-acylated zinc complex, that is to say the complex of zinc cations with the

  
  <EMI ID = 97.1>

  
béré by the destruction of the complexing association in

  
  <EMI ID = 98.1>

  
substantial absence of the organic solvent medium, (c) of

  
  <EMI ID = 99.1>

  
truction of the complexing association in the part of the N-acylated zinc complex, (d) an amount of the zinc salt which was initially added in excess and which did not react during the complexing reaction, and optionally (e) a certain rent of the organic solvent used during the preceding operations.

  
The aforementioned oily or solid deposit or residue (ci

  
  <EMI ID = 100.1>

  
hard (a), (b) and (c) below.

  
  <EMI ID = 101.1>

  
that polar, aqueous or not, alone or mixed with others, of the type of polar organic liquids which have as ef-

  
  <EMI ID = 102.1>

  
zinc in the N-acylated complex present in the said deposit or reacted and in which the quantities of zinc salt released and those initially present and which have not reacted are soluble, but which do not dissolve the desired N-acylated derivative of the aminoglycoside antibiotic. In this way, the N-acylated zinc complex is destroyed to release the cations of this metal and allow them

  
to dissolve in the form of zinc salt in water or the (a) organic solvent (.), aqueous or not, abovementioned, leaving the desired N-acylated derivative of antibiotic

  
  <EMI ID = 103.1>

  
recover. This residue can be optionally purified

  
by dissolving it again in an organic solvent. The polar organic solvents available in this procedure (a) are, for example, methanol, ethanol, ammonia, ethylamine and triethylamine. Consequently, these polar organic solvents and water serve as a reagent for the removal of zinc cations.

  
(b) As a variant, the deposit can be mixed or <EMI ID = 104.1>

  
type of polar organic solvent, either anhydrous or aqueous, which has the effect of destroying the complexing association of zinc cations in the N-acylated complex of this metal which is present in said deposit or residue, and in which the salt of released zinc is not soluble, while the desired N-acylated derivative of aminoglycoside antibiotic

  
  <EMI ID = 105.1>

  
acylated is destroyed to release the aforementioned derivative and allow it to dissolve in the said polar organic solvent and to be extracted therefrom and, by summoning, to be. 'by' zinc salt, which is released, but rent <EMI ID = 106.1>

  
air of aminoglycoside antibiotic in the polar organic solvent and, if desired, it can be purified,

  
  <EMI ID = 107.1>

  
purified to isolate the derivative in question.

  
(c) As another variant, it is possible to dissolve the oily or solid deposit or residue (the aforementioned mixture) obtained by the second method described above, as it is, in an appropriate organic solvent containing a proportion of water, if the all of the deposit or residue is soluble in water, at least to a substantial extent.

  
The solution thus obtained can be subjected to a chroma graphic treatment during which the liberated zinc salt and the liberated N-acylated derivative can be extracted separately from the solution. The authors of the present invention have found that various kinds of cationic resins, anionic resins, chelate exchange resins and water-insoluble high polymers which contain functional groups capable of being used for this chromatographic treatment. to combine with a metal, such as chitin or chitosan. Among the kinds of cationic resins which can be used for this purpose, mention may be made of those which contain carboxylic groups (-COOH) as active groups and those in which the functional groups d are provided by

  
  <EMI ID = 108.1>

  
active groups, the oily or solid deposit or residue (the aforementioned mixture) is dissolved in a suitable aqueous organic solvent, for example a mixture of water and

  
  <EMI ID = 109.1> <EMI ID = 110.1>

  
another amount of this solvent further containing a proportion of an acid or a base. In the case of an acid, a weak organic acid, such as acetic acid, or a dilute inorganic acid, such as

  
  <EMI ID = 111.1>

  
ammonium hydroxide can be used in most cases. The concentration of acid or base in the developing solvent (the eluent) can be understood

  
  <EMI ID = 112.1>

  
  <EMI ID = 113.1>

  
complexing zinc cations during the development process, because the cationic resin used has different adaorptive affinities with respect to the N-acylated aminoglycoaide and zinc cations, aorta that the force which pushes the first to * associate with the resin is different from that which tends to associate these

  
  <EMI ID = 114.1>

  
  <EMI ID = 115.1>

  
  <EMI ID = 116.1> <EMI ID = 117.1>

  
ion ion between them, so that the development of the column of anionic resin with a suitable aqueous organic solvent makes it possible to elute out of the column the

  
  <EMI ID = 118.1>

  
that the zinc cations remain in the last coat.

  
When the chromatographic process is carried out using a chelate exchange resin, which can combine with zinc cations thanks to the ability it

  
a to form a chelate with metals, we place a solution

  
  <EMI ID = 119.1>

  
tee) in an aqueous organic solvent in a column of the chelate exchange resin and then developed

  
  <EMI ID = 120.1>

  
  <EMI ID = 121.1>

  
ment at the bottom of the column, while the zinc cations remain attached to the resin. The water-insoluble high polymer, which contains functional groups capable of combining with a metal, such as chitin and chitosan for example, can be used in the same manner as when using the chelate exchange resin.

  
(d) It is also possible to use a third method, in which a cationic resin, anionic resin, chelate exchange resin or a water-insoluble high polymer containing functional groups capable of <EMI ID = 122.1>

  
acylation carried out to protect the amino groups, so that the N-acylated zinc complex is adsorbed by the resin or the high polymer. The column can then be washed with an aqueous organic solvent, if necessary, and then developed with an aqueous organic solvent containing or not an acid or a base, as in method (c), and the elimination is thus obtained. zinc cations of the N-acylated complex as well as recovery of the desired N-acylated derivative of aminoglycoside antibiotic.

  
(e) There is also a fourth method for re- <EMI ID = 123.1>

  
cosidique, method in which the reaction mixture containing the N-acylated zinc complex is treated immediately with water by incorporating it into water in the case where the said desired derivative itself is insoluble, at least substantially, in the latter. As an example, an N-acylated derivative insoluble in water can be cited.

  
  <EMI ID = 124.1>

  
zyloxycarbonyldibékacino. In this case, when the acylation reaction mixture containing the Nacylated zinc complex comprises an N-acylated derivative substantially insoluble in water of an aminoglycoside antibiotic is immediately incorporated into water, the complexing association of zinc in the N-acylated complex is destroyed to allow the insoluble N-acylated derivative to precipitate us in the form of a solid, while the zinc salt formed by the cations released from this metal remain in solution, so that the derivative N-acylated antibiotic desired

  
  <EMI ID = 125.1>

  
of zinc in the form of a practically pure product.

  
As explained above, N-acylation, i.e. the amino group protection reaction, is carried out with the zinc complex of the antibiotic

  
  <EMI ID = 126.1>

  
a complex in which the zinc cations used are associated in the form of a complex with the structure of the

  
  <EMI ID = 127.1>

  
highly soluble in water, a simple operation which consists in incorporating water into the mixture of the acylation reaction containing the N-acylated zinc complex causes the precipitation of the N-acylated derivative insoluble in water under the form of a solid, while the released zinc cations are eliminated by dissolution in water (com-

  
  <EMI ID = 128.1> <EMI ID = 129.1>

  
partially soluble in water and can therefore only be collected with a considerably reduced yield

  
  <EMI ID = 130.1>

  
Immediately add water to the acylate reaction mixture. For these reasons, better results are obtained.

  
  <EMI ID = 131.1>

  
res (b) and (c), in which the Nacylated zinc complex (i.e. the zinc cation complex with the N-acylated derivative of aminoglycoside antibiotic resulting from the acylation reaction) is all first separated from the reaction mixture and then dissolved in

  
  <EMI ID = 132.1>

  
lution thus obtained is treated to remove the zinc cations it contains. One of the simple methods of removing the obvious zinc cations for now is that in which hydrogen sulfide or an alkali sulfide is reacted as a precipitation agent with the zinc cations to precipitate the latter. in the form of zinc sulfide (form of implementation of the first method described in paragraph (a)). However, zinc sulfide often precipitates in the form of a colloidal deposit which is difficult to separate by filtration and hydrogen sulfide or alkaline sulfide also have an unpleasant odor and are not suitable for application of the process. on an industrial scale.

   The authors of the invention therefore carried out extensive research with a view to providing a practical method of removing zinc cations from the complex without resorting to the intervention of a sulfide and they therefore succeeded in developing the effical methods and easy to remove which consists in using the resins <EMI ID = 133.1>

  
case of procedures (c) and (d)). These procedures (c) and (d) are industrially very advantageous and advantageous, because they are easy to carry out, have a high efficiency from the point of view of the separation of zinc cations and provide a high yield of the N derivative. -desired acylated aminoglycosidic antibiotic.

  
The various methods and procedures for treating the N-acylated zinc complex with the elimination reagent which have been described above can be summarized as follows:
(a) The zinc cation complex with the derivative <EMI ID = 134.1>

  
  <EMI ID = 135.1>

  
  <EMI ID = 136.1>

  
tions of zinc it contains.

  
(b) The complex of zinc cations with the derivative <EMI ID = 137.1>

  
is separated from the acylation reaction mixture by extraction with an organic solvent, by evaporation of the organic solvent medium contained in the reaction mixture or by dilution of this mixture with an organic solvent

  
  <EMI ID = 138.1>

  
elimination of zinc cations.

  
(c) The complex of zinc cations with the derivative <EMI ID = 139.1>

  
eat, once separated. mixed with water or a polar organic solvent, anhydrous or aqueous, which serves as a reagent for removing zinc cations. This polar organic solvent is either a solvent in which the zinc salt is soluble but which does not dissolve the N-acyl derivative of aminoglycoside antibiotic, or a solvent in which the zinc salt is insoluble, but which dissolves the derivative N -acylated aminoglycosidic antibiotic.

  
(d) The complex of zinc cations with the N'acylated derivative of aminoglycosidic antibiotic is, once separated, again put to dissolve entirely in an inorganic solvent containing a proportion of water and the resulting solution is subjected chromatographic treatment using a cationic resin, a

  
  <EMI ID = 140.1>

  
water-insoluble high polymer containing functional groups capable of combining with a metal, which serve as a reaction for the elimination of zinc cations.

  
(e) The mixture of the acylation reaction is passed directly through a column of a cationic resin, an anionic resin, a chelate exchange resin or a water-insoluble high polymer. and containing functional groups capable of combining with a metal to obtain the adsorption of the zinc cation complex with the N-acylated derivative of antibiotic <EMI ID = 141.1>

  
aqueous organic solvent containing or not a proportion of an acid and a base, and the eluate is collected in fractions, operation followed by recovery of the fractions which contain the selectively N-acylated derivative

  
  <EMI ID = 142.1>

  
do not have zinc cations.

  
(f) When the desired N-acylated derivative of antibiotic <EMI ID = 143.1>

  
soluble in water, the mixture is immediately incorporated

  
  <EMI ID = 144.1>

  
  <EMI ID = 145.1>

  
both dissolved in water.

  
(g) The mixture of the acylation reaction is immediately treated with hydrogen sulfide, an alkali or alkaline earth metal sulfide which precipitates the zinc cations in the form of the sulfide of this "lethal, or with ammonium hydroxide, which precipitates them in the form of groin hydroxide.

  
In the zinc complex formed in the process according to the first aspect of the invention, the mine cations

  
  <EMI ID = 146.1> such that the N-acylation of the aminoglycoside antibiotic complex

  
  <EMI ID = 147.1>

  
cations, normally produces the N-acyl derivative in which amino and / or alkylamino groups other than those occupying

  
  <EMI ID = 148.1>

  
When the N-acylated derivative of aminoglycoaidic antibiotic

  
  <EMI ID = 149.1>

  
  <EMI ID = 150.1> example, operation followed by the elimination of the residual amino-protective groups from the N-acylated product which

  
  <EMI ID = 151.1>

  
1-N-acylated synthetic, which is known to be a useful antibacterial agent.

  
  <EMI ID = 152.1>

  
1-N-acylea is now described with reference to the use (non-limiting) of kanamycin A as a starting material. When, in the process according to the first aspect of the invention, kanamycin A is used as the

  
  <EMI ID = 153.1>

  
Kanamycin are initially blocked by the formation of a complex with zinc cations. Consequently, when the kanamycin A-zinc zinc complex is acylated with an appropriate acylation reagent according to the invention or with

  
  <EMI ID = 154.1>

  
of kanamycin A can be protected by the acyl group of the acylating reagent used or by the other type of blocking agent. After removal of the complexing zinc cations from the N-acylated kanamycin A complex with the zinc cations, the resulting N-acylated kanamycin A derivative is reacted with an acylating agent having

  
  <EMI ID = 155.1>

  
kanamycin A. At this point the 1-amino group is normal- <EMI ID = 156.1>

  
with a yield slightly higher than that of the 3 "-N-acylated derivative of this tiny kanamycin. The subsequent elimination of the amino-protecting groups of the 1-N-acylated derivative thus obtained gives kanamycin A 1-N-acylated as final product It is therefore easily understood that when the method according to the first aspect of the invention is used, the desired 1-N-acylated kanamycin A can be obtained in a higher yield than if the non-kanamycin A is reacted. protected or kanamycin A protected in 6 'immediately with an acylating agent to achieve

  
the 1-N-acylation of kanamycin A. If a non-amino-protected kanamycin is reacted with a 1-Nacylating agent, it is found that a mixture of N-acylea products is formed which contains a very small proportion (habi-

  
  <EMI ID = 157.1>

  
re.

  
In the case where the method according to the first aspect of the invention is applied to a kanamycin of general formula (III), some or all of the amino groups other than the 1- and 3 "-amino groups of this kanamycin are protected to give a derivative of kanamyci-

  
  <EMI ID = 158.1>
  <EMI ID = 159.1>
  <EMI ID = 160.1> <EMI ID = 161.1>

  
blocked. However, if the acyl group to be introduced as an amino-blocking group has a relatively large steric size, for example the t-butoxycarbonyl group, or has the molar amount of the acylation reagent used

  
  <EMI ID = 162.1>

  
necessary for acylor all the uncomplexed amino groups of the kanamycin molecule, even if the acyl group of the acylation reagent used has an ordinary size, or if the acylation reaction is stopped at an intermediate stage, a derivative is obtained N-protected from kanamycin in which the number of acylated amino groups in the kanamycin molecule is less than it is

  
  <EMI ID = 163.1>

  
is exclusively acylated, since this 6'amino or 6'-alkylamino group is more reactive than the other amino groups of the kanamycin molecule.

  
The N-acylated kanamycin derivative of general formula
(V) is an important intermediate product, useful in the semi-aynthetic preparation of various types of kanamycin derivatives. The compound of formula (V) has a greater value as an intermediate substance for chemical synthesis, for example, in particular when it takes part in a process for the preparation of semi-synthetic 1-N-acylated aminoglycosidic antibiotics, active against kanamycin-resistant bacilli, by acylation of

  
  <EMI ID = 164.1>

  
hydroxyl-u-aminoalkanotque, then elimination of the protective groups of the blocked amino and / or alkylamino groups of the 1-N-acylation product obtained.

  
For example, when the intermediate compound of formula (V) is to be acylated with an acyl group, such as <EMI ID = 165.1>

  
an a &#65533; butyric ido substituted in consequence or its derivative

  
  <EMI ID = 166.1>

  
N-hydroxyauccinimide ester, N-hydroxyphthalimide ester or p-nitrophenol ester, which has the effect of forming the product of 1-N-acylation. Then, the elimination of the benzyloycarbonyl group and of the protective group
(R) in formula (V) the 1-N-acylation product can be performed by applying a traditional technique

  
  <EMI ID = 167.1>

  
hydrolysis with an acid or a base, either by reduction by means of a reducing metal, or again by catalytic hydrogenolysis with hydrogen, or finally by radical reduction with sodium in liquid ammonia, to give a derivative semi-synthetic of kanamycino having a group (S) -4-amino-2-hydroxybutyryl linked to the 1-amino group of kanamycin, derivative which is active with regard to resistant bacilli and is represented by the general formula (VI ) below:

  

  <EMI ID = 168.1>
 

  
  <EMI ID = 169.1>

  
in which R, R, R, R each have the same definition as in formula (III). In the procedure below. above, we

  
  <EMI ID = 170.1>

  

  <EMI ID = 171.1>


  
  <EMI ID = 172.1>

  
  <EMI ID = 173.1>

  
kanamycin.

  
The invention further embraces a process for the preparation of an N-acylated derivative selectively protected from an aminoglycoside antibiotic comprising a 6-0- moiety

  
  <EMI ID = 174.1>

  
atreptamino possibly having a group 4-0- (aminoglycoaylo), derivative in which all the amino groups (including

  
  <EMI ID = 175.1>

  
identical or different acyls.

  
By the method according to the first aspect of the invention (sometimes called the "zinc complex" method in the remainder of this document), it is possible to prepare an N-acylated derivative, selectively but partially protected from the aminoglycoside antibiotic, derivative in which all amino groups other than the two 1-amino groups

  
  <EMI ID = 176.1>

  
reactive equivalent to carry out the 1-N-acylation described above, it is in fact that a mixture of acylation products is obtained comprising (a) the 1-N-acylated product in which only the 1-amino group of the aminoglyco- molecule

  
  <EMI ID = 177.1>
(b) the 3 "-N-acylated product, where only the 3" -amino group (or <EMI ID = 178.1>

  
has not been acylated. To obtain the desired final 1-N-acylation product from the above mixture, it is therefore always necessary to carry out an additional operation, in which the final 1-N-acylation product is isolated from the mixture by chromatography or by another isolation process. How happy the 1-amino group is-

  
  <EMI ID = 179.1>

  
amino and 3 "-amino (or 3" -alkylamino) have identical reactivity. However, even if the 1-N-acylation reaction is carried out under optimal conditions, it is inevitable that undesirable N-acylated products will also form and therefore an additional operation is always necessary.

  
  <EMI ID = 180.1>

  
  <EMI ID = 181.1>

  
mixed product by column chromatography.

  
To eliminate this drawback, it is obviously necessary to prepare a selective N-acyl derivative.

  
  <EMI ID = 182.1>

  
which all amino groups other than the 1-amino group have been protected. In order for this condition to be fulfilled, the authors of the invention have carried out other research with a view to providing a process which makes it possible to protect

  
  <EMI ID = 183.1>

  
the N-acylated and selectively but partially protected aminoglycoside antibiotic derivative which, when prepared according to the "zinc complex" process,

  
  <EMI ID = 184.1>

  
this protection without blocking the 1-amino group.

  
This research led the authors of the invention to discover that when the partial N-acylated derivative

  
  <EMI ID = 185.1>

  
the “zinc complex” process is reacted with an acylating agent chosen from esters of fornic acid,

  
  <EMI ID = 186.1>

  
can be preferably acylated for blocking purposes without acylation of the 1-amino group. This 3 "-N-selective protection process can be combined with the" zinc complex "process described above (that is to say the process according to the first aspect of the invention), which makes it possible to produce d conveniently and effectively the selectively protected N-acyl derivative of the antibiotic

  
  <EMI ID = 187.1>

  
selectively protected by identical or different acyl groups. This combination of the "zinc complex" process with the 3 "-N-.elective protection of- process

  
  <EMI ID = 188.1>

  
see more. When the abovementioned derivative is used for the 1-N-acylation of the aminoglycoside antibiotic, there is the additional advantage that it practically does not form undesirable N-acylated by-products, which considerably facilitates the separation and purification of the desired 1-N-acylation product "

  
According to the second aspect of the present invention, there is therefore provided a process for the preparation of an N-acylated derivative selectively protected from an aminogly-

  
  <EMI ID = 189.1> <EMI ID = 190.1>

  
by identical or different acyl groups, process which consists in reacting an alkanotque acid ester of formula (VIII):

  

  <EMI ID = 191.1>


  
in which R is a hydrogen atom or a group

  
  <EMI ID = 192.1>

  
  <EMI ID = 193.1>

  
6 carbon atoms, an aralkyloxy group, especially the benzyloxy group, an aryloxy group, especially the phenyloxy group, or an N-formylimidazole, as acylating agent, in an inert organic solvent, with an N-acylated derivative partially protected from aminogly antibiotic

  
  <EMI ID = 194.1>

  
3 "-alkylamino) are not protected, all other amino groups being protected by an acyl group as amino protecting group, to effect selective acylation of

  
  <EMI ID = 195.1>

  
acylating and thus give the desired derivative, entirely Nprotected jump in position 1, of the antibiotic aminoglycoaidique.

  
  <EMI ID = 196.1>

  
be used in the process according to the second aspect of the invention are the same as those which have been mentioned during the description of the process according to the first aspect of the invention.

  
Forms of mine implementing the method according to the second aspect of the invention are now described more fully.

  
The partially protected N-acylated derivative of antibiotic

  
  <EMI ID = 197.1>

  
  <EMI ID = 198.1>

  
invention and wherein all other amino groups

  
  <EMI ID = 199.1> <EMI ID = 200.1>

  
lon the first aspect of the invention. Consequently, the acyl group initially present in the N-acylated derivative

  
  <EMI ID = 201.1>

  
partially protected N-acylated derivative of aminoglycoside antibiotic used as starting material can also be a derivative prepared according to the Nagabhushan process

  
  <EMI ID = 202.1>

  
In the mine implementing the method according to the second aspect of the invention, the partially protected N-acylated derivative of aminoglycoside antibiotic in which the

  
  <EMI ID = 203.1>

  
derivative used. The inert organic solvent can be

  
preferably one of those with a high dissolving power with respect to the starting material, for example

  
  <EMI ID = 204.1>

  
n-butanol and t-butanol, as well as the aqueous forms <EMI ID = 205.1>

  
although they are not very suitable, giving a lower yield of the desired product. With the acylating agent of formula (VIII), Ra may preferably be a dihaloalkyl or trihaloalkyl group, in particular the

  
  <EMI ID = 206.1>

  
le, and R may preferably be an alkyloxy group, such as the methoxy or ethoxy group. When R is an aryloxy group, it can be the ph & noxy group. As examples of the acylating agent of formula (VIII), mention may in particular be made of methyl formate, ethyl formate,

  
  <EMI ID = 207.1>

  
phenyl, methyl dichloroacetate, methyl trichloroacetate, phenyl trichloroacetate, methyl trifluoracetate, ethyl tritluoracetate and

  
  <EMI ID = 208.1>

  
of acylating agent, it is possible to formylate, dichloracetyler, trichloracetyler or trifluoroacetyler preferentially

  
  <EMI ID = 209.1>

  
cile to be removed during the subsequent phase of N-deprotection by a conventional method of removing protective groups. If an ester of alkanol acid and alkanol of form is not used as the acylating agent

  
  <EMI ID = 210.1>

  
that or an active ester thereof, such as the ester of Nhydroxyauccinimide (which is not in accordance with the second aspect of the invention), acylation cannot be obtained

  
  <EMI ID = 211.1>

  
in particular a 1-N-acylated product and / or a mixed acylation product which contains the 1-N-acylated product. It is remarkable that one cannot obtain selective acylation rock-

  
  <EMI ID = 212.1>

  
cylant an anhydride or an active ester of the same acid al-canolquo.

  
The acylating agents of formula (VIII) which can be

  
  <EMI ID = 213.1>

  
The invention have various reactivities, ranging from a wide range between "strong" and "weak". When an acylating agent having a high reactivity is used, the acylation reaction can take place over a short period while cooling. On the other hand, when an acylating agent having a weak reactivity is used, the acylation reaction can be carried out either by heating, or over a longer period of time. In general, however, the suitable reaction temperature can be between -30 ° C and + 120 ° C and the suitable reaction time can be between 30 minutes and 24 hours, even 48 hours.

  
The desired selectively 3 "-N-acylated product can be isolated from the reaction mixture in a known manner, for example by evaporation of the solvent or by precipitation with addition of water, if necessary, followed by purification of the product.

  
  <EMI ID = 214.1>

  
Selective tion in the method according to this second aspect of the invention is not yet fully explained. One possible interpretation is that the ncylating agent of formula (VIII) first acylates a hydroxyl group from the starting product to form an ester as an intermediate product and then this 0-esterifying acyl group is displaced or emigrated to a group. amino (in this case-

  
  <EMI ID = 215.1>

  
is close to the intermediate esterified hydroxyl, which causes the acylation of this amino group. If this hypothesis is adopted, it is possible to understand why the 1-amino group, which does not border on any hydroxyl group, cannot be acylated in the process according to the second aspect of the invention. Moreover, it is a fact that this intermediate ester cannot be obtained when the trifluoroacetylation or the formylation is con- <EMI ID = 216.1>

  
O-formyl is unstable and that a quantity of this unstable acyl group which has not undergone group migration

  
  <EMI ID = 217.1>

  
tion and purification of the product of 3 "-N-acylation so as to restore the free hydroxyl group. The invention

  
  <EMI ID = 218.1>

  
  <EMI ID = 219.1>

  
in the process according to the second aspect of the invention, those which are most suitable are those which have an acyl group capable of giving a more unstable ester when it is transformed into O-acyl group by reaction with the hydroxyl group and thus gives an ester product. It is meanwhile very interesting to note that when the method according to the second aspect of the invention is implemented using instead of N-fortnylimidazo-

  
  <EMI ID = 220.1>

  
protected from the aminoglycoside antibiotic is not a-

  
  <EMI ID = 221.1>

  
N-alkanoylimidazole alkanoyl used To give an intermediate product of O-esterification. When this pro-

  
  <EMI ID = 222.1>

  
This product is then treated with an alkaline reagent, such as ammonium hydroxide, at room temperature.

  
  <EMI ID = 223.1> <EMI ID = 224.1>

  
N-acylated desired in the reaction mixture of the partially protected N-acylated derivative of the aminoglyoside antibiotic with an N-alkanoylimidazole, but it cannot be

  
  <EMI ID = 225.1>

  
made alkaline by treating it with an alkaline reagent such as ammonia (see Example 71).

  
As an interesting application of the methods according to the first and second aspect of the invention, it is possible to provide a method of high yield for the preparation of the aminoglycoside antibiotic 1-N-

  
  <EMI ID = 226.1>

  
noalcanoyl) aminoglycoside starting from the parent aminoglycoside antibiotic, a process which consists of a combination of the preparation, using the process according to the first aspect of the invention, known as the "zinc complex, a partially protected N-acylated derivative of the starting aminoglycoside antibiotic, in which the 1-amino and 3 "-amino or 3" -alkylamino groups are not protected and all the other amino groups are protected,

  
  <EMI ID = 227.1>

  
tive according to the second aspect of the invention, of the derivative

  
  <EMI ID = 228.1>

  
of the 1-amino group, of the acylation of the 1-amino group of

  
  <EMI ID = 229.1>

  
(isoserine) or 4-amino-2-hydroxybutyric acid, and finally the elimination of the protective groups in the 1-N-acylation product thus obtained.

  
  <EMI ID = 230.1>

  
invention provides a better method of preparing a

  
  <EMI ID = 231.1>

  
optionally having a 4-0- (aminoglycosyl) group, said process consisting in:
(a) reacting zinc cations with the starting aminoglycoside antibiotic in an inert organic solvent to produce the complex of these zinc cations with this aminoglycoside antibiotic;
(b) reacting an acylation reagent having an acyl group to be introduced as an amino-protective group <EMI ID = 232.1>

  
of zinc formed in the previous step (a), in situ in the inert organic solvent, to produce a complex of zinc cations with the selectively N-acylated derivative of the aminoglycoside antibiotic. derivative in which

  
  <EMI ID = 233.1>
(c) reacting the complex N-acylated derivative of the aminoglycoside antibiotic-zinc cation8 obtained during step (b) above with a reagent which removes zinc cations from the N-acylated zinc complex, for obtain a partially and selectively protected N-acylated derivative of the aminoglycoside antibiotic, which is free of zinc cations and in <EMI ID = 234.1>

  
are not protected, all other amino groups of the aminoglycoside molecule being protected by the acyl group;
(d) reacting the N-acylated derivative, partially <EMI ID = 235.1>

  
dique obtained during step (c) above with an alkanoic acid ester of formula (VIII):

  

  <EMI ID = 236.1>


  
in which R is a hydrogen atom or a dihaloalkyl or trihaloalkyl group containing from 1 to 6 carbon atoms and Rb is an alkyloxy group containing do 1 to

  
6 carbon atomea, an aralkyloxy group containing 1

  
with 6 carbon atoms in its alkyl fraction, in particular the benzyloxy group, or an aryloxy group, in particular the phenoxy group, or N-formylimidazole, such as <EMI ID = 237.1>

  
and thus give the derivative of the aminoglycoside antibiotic in which all the amino groups other than the 1-amino group are protected by the acyl group,
(e) reacting the fully N-protected derivative except in position 1 obtained during step (d) above <EMI ID = 238.1>

  

  <EMI ID = 239.1>


  
in which m is 1 or 2, or an equivalent reactive derivative, in which the amino group is protected or not, in order to acylate the 1-amino group of the abovementioned antibiotic derivative, and <EMI ID = 240.1> <EMI ID = 241.1>

  
previous step (e) by a traditional disposal method.

  
A more complete description is now given of how to implement the method according to this third aspect of the invention.

  
  <EMI ID = 242.1>

  
be used as starting antibiotic in the first step of this process are the same as those indicated in the description of the process according to the first aspect of the invention and the reaction which produces the cation complex

  
  <EMI ID = 243.1>

  
thus obtained in (a) can only be carried out in the second step (b) of the present method in the same way as that described for the method according to the first aspect of the invention. Elimination of zinc cations from the antibiotic complex

  
  <EMI ID = 244.1>

  
thus obtained can be carried out, during the third step (c) of the present process, in various ways already described and makes it possible to obtain an N-acylated derivative, partially and selectively protected, of the aminoglycosidic antibiotic. which is free from zinc cations and in which the 1-amino and 3 "-amino or 3" -alkylamino groups are not protected, but all other amino groups in the aminoglycoside molecule are blocked by the acyl group of the reagent acylation used in step (b) of the present process. This N-acylated derivative, partially and se-

  
  <EMI ID = 245.1>

  
is then reacted with an ester of alkanoic acid

  
  <EMI ID = 246.1>

  
step (d) of the present method in the same manner as that already described with regard to the method according to the second aspect of the invention, in order to obtain the selective 3 "-N-acylation of the partially N-protected derivative of the aminoglycoside antibiotic without acylation of the 1-amino group it contains.

  
During the fifth step (e) of the present process, the fully protected derivative except in position 1 obtained during the previous step (d) is reacted

  
  <EMI ID = 247.1>

  
droxybutyryl. This 1-N-acylation can be carried out in a general manner like that described in the patent.

  
  <EMI ID = 248.1>

  
before any known method of amide synthesis, by reaction of the protected derivative of the antibiotic a-

  
  <EMI ID = 249.1>

  
2-hydroxybutyric acid, either in its free acid form or in that of its reactive equivalent, such as an ester

  
  <EMI ID = 250.1>

  
its mixed anhydride, its azide, in an inert organic solvent, such as dioxane, dimethoxyethane, <EMI ID = 251.1>

  
aqueous of these solvents. Isoserine and L-4-amino-2-hydroxybutyric acid may have their amino group blocked by an amino protecting group. The amino protecting group suitable for this purpose may be the same or different from that used in the fully N- derivative.

  
  <EMI ID = 252.1>

  
preferred protector because it can be easily removed by treating with a dilute acid such as aqueous trifluoroacetic acid, aqueous acetic acid and dilute hydrochloric acid. The benzyloxycarbonyl group, which is eliminated by traditional catalytic hydrogenolysis using palladium or platinum oxide as catalyst, as well as the phthaloyl group, which is easily eliminated by hydrolysis with hydrazine, are perfectly suitable as amino protecting group for this purpose.

  
The acylation reaction in the 1-N-acylation step (e) of the process according to the third aspect of the invention can preferably be carried out in a solvent or-

  
  <EMI ID = 253.1>

  
remove the 1-N-acylation product obtained in step (e) from all of its amino-protecting groups. This elimination can only be done by applying a traditional method.

  
  <EMI ID = 254.1>

  
be removed by hydrolysis with an aqueous solution of <EMI ID = 255.1>

  
dilute acid solution, such as dilute hydrochloric acid. A residual amino-protecting group of aralkyloxycar- type

  
  <EMI ID = 256.1>

  
eliminated by traditional catalytic hydrogenolyae. When all of the residual amino protecting groups are

  
  <EMI ID = 257.1>

  
minoalcanoyl) -aminoglycoaidique prepared by the process according to this third aspect of the invention are listed. below:

  
(1) 1-N- (L-4-amino-2hydroxybutyryl) -kanamycin A

  
  <EMI ID = 258.1>

  
Another application of the methods according to the first and second aspect of the invention consists in pre-

  
  <EMI ID = 259.1>

  
that the 1-amino group is not protected, and as an example of this application, mention may be made of the preparation

  
  <EMI ID = 260.1>

  
laughing.

  
  <EMI ID = 261.1>

  
following examples, which in no way constitute a limitation.

Example 1

  
  <EMI ID = 262.1> <EMI ID = 263.1>

  
then stirred at room temperature until the reaction mixture forms a homogeneous solution. It takes about 4-5 hours for a zinc do kanamy complex to

  
  <EMI ID = 264.1>

  
The reaction solution is kept at room temperature for 4 hours, during which the zinc complex of kanamycin A undergoes a bonzyloxycarbonylation.
(acylation in accordance with the first aspect of the invention).

  
A sample taken from the reaction solution thus obtained is subjected to chromatography.

  
thin layer on silica gel using, as development solvent the lower liquid phase of a

  
  <EMI ID = 265.1>

  
to byproducts with higher back points.

  
(ii) The above reaction solution is poured into
500 ml of ethyl ether and the separated oil is washed several times with other quantities of ethyl ether to give 8.8 g of a thick syrupy material.
(iii) The zinc cation is removed from the syrupy material (essentially comprising the zinc complex) according to one or the other of the following methods.

  
  <EMI ID = 266.1> <EMI ID = 267.1>

  
ninhydrin is first removed from the column by elution and zinc acetate, sensitive to coloration by diphenylcarbnzide, is then removed by elution. The fractions containing the desired product are combined and concentrated to dryness. The residue is washed with ether

  
  <EMI ID = 268.1>

  
B - Processes using a weak cation exchange resin carrying a carboxylate group as functional group (sold under the name of "Amberlito CG 50" (form

  
  <EMI ID = 269.1>

  
the solution is passed through a column of 60 ml of Amberlito CG 50 resin (NH4 + form); it is subjected to an elution by linear gradients with water-dioxane
(1: 1) with ammonia concentrations ranging from 0

  
  <EMI ID = 270.1>

  
  <EMI ID = 271.1>

  
  <EMI ID = 272.1>

  
C - Process using a cation exchange resin

  
  <EMI ID = 273.1>

  
then an elution is made by linear gradients with water-dioxane (2: 1) with ammonia concentrations ranging from 0 to IN. The eluate fractions containing the

  
  <EMI ID = 274.1>

  
of a white solid identical to that obtained in Example 1 (iii) (B).

  
D - Another process using the Dowex 50W X 2.

  
Passed through a 30 ml column of Dowex

  
  <EMI ID = 275.1>

  
obtained in Example I (ii) in 20 ml of water-methanol
(3: 1). The column is washed well with water-methanol
(3: 1) then an elution is carried out by gradients with water-methanol with C1H concentration * of 0 to 6 N. The active fractions containing the desired 3,6'-di-N-benzyloxycarbonylkanamycin A are combined and mixed with a strongly basic anion exchange resin
(OH form) in sufficient quantity to make the mixture slightly acidic.

  
The mixture is filtered, and the filtrate concentrated

  
  <EMI ID = 276.1> E - Process using an anion exchange resin carrying a highly basic quaternary ammorium functional group (sold under the name "DOWEX 1 X 2" by Dow Chemical).

  
  <EMI ID = 277.1>

  
created in a column of 30 ml of Dowex 1 X 2 resin
(OH form) previously impregnated with water-dioxane
(1: 1) then the column is developed at relatively high speed with water-dioxane (1: 1). The eluate fractions containing the desired product are collected

  
  <EMI ID = 278.1>

  
colorless solid identical to that of Example 1 (iii) B. F - Process using an anion exchange resin carrying a weakly basic functional group (sold under the name "Dowex WGR" by Dow Chemical C *).

  
1 g of the syrupy material obtained in Example 1 (ii) is dissolved in 20 ml of water-dioxane (2: 1) and the solution is passed through a column of 50 ml of Dowex WGR (base form) beforehand saturated with dioxane water (2: 1) and then eluted with dioxane water (2: 1). The 3,6'-di-N-benzyloxycarbonylkanamycin A is eliminated by elution in a few fractions, with entrainment of a trace of zinc. These fractions are combined and concentrated to dryness to give 450 mg of a colorless solid. The solid can be used directly as a material

  
starting material for the production of l-N ((S) -4-amino-2-hydroxybutyryl) kanamycin A according to the 1-N acylation method of Example 31 below, in which the traces

  
of zinc cation remaining in the starting solid do not exert an adverse influence on the acylation reaction

  
  <EMI ID = 279.1>

  
G - Process using an exchange-chelate resin carrying a weakly acidic functional group (sold under the name

  
  <EMI ID = 280.1>

  
A solution of 1 g of the syrupy material, obtained in Example 1 (II), is introduced into water. <EMI ID = 281.1>

  
ées that in a later phase as effluent from the column are combined * and concentrated to dryness to give 272 mg (74%) of the desired product as a white solid.

  
H - Process using Chitosan (polymer insoluble in water containing functional groups susceptible

  
  <EMI ID = 282.1>

  
Carefully impregnate 100 ml of Chitosan with water-methanol (3: 1) and fill a column in which a solution of 1 g of the

  
  <EMI ID = 283.1>

  
fraction * sensitive * to the reaction to ninhydrin are

  
  <EMI ID = 284.1>

  
  <EMI ID = 285.1>

  
Example 1 (111) (B).

  
1 - Process using a high polymer carrying holes

  
  <EMI ID = 286.1> <EMI ID = 287.1>

  
Example 1 (iii) (B).

  
J - Process using hydrogen sulfide to precipitate zinc.

  
1 g of the syrupy material obtained in Example 1 (ii) is dissolved in 20 ml of water-methanol (1: 1),

  
aqueous ammonia is added thereto, then a sufficient quantity of hydrogen sulphide is introduced. The reaction mixture containing the zinc sulfide precipitate is filtered through a glass filter filled with the "Celite" filter aid and the filtrate is concentrated at reduced pressure to give a syrupy material which is well

  
  <EMI ID = 288.1>

  
  <EMI ID = 289.1>

  
and the solution is chromatographed on a column of
30 ml of Amberlite IRA 900 (forma OH, strongly ba-

  
  <EMI ID = 290.1>

  
him from Example 1 (iii) (B).

Example 2

  
  <EMI ID = 291.1> <EMI ID = 292.1>

  
840 mg (6.18 moles) sodium acetate trihydrate. After stirring the mixture at room temperature for 4 hours, a solution of 675 mg (2.27 moles) of N-benzyloxycarbonyloxyphthalimide is slowly added over about an hour to the mixture containing the kanamycin A-zinc complex formed over room temperature for 4 hours.

  

  <EMI ID = 293.1>


  
of dimethyl sulfoxide. The mixture obtained is left to stand at room temperature for 4 hours.

  
The reaction mixture is then treated with the ma-

  
  <EMI ID = 294.1>

  
cine A is a colorless solid.

Example 3

  
  <EMI ID = 295.1> 1 g (4.55 mole *) of zinc acetate dihydrate is added, followed by continuous stirring for an additional 5 hours. We add in 30 minutes to the mixture

  
  <EMI ID = 296.1>

  
hydroturan (1: 1). After the mixture obtained has been stirred overnight at room temperature,

  
  <EMI ID = 297.1>

  
tyle forms us of precipitate. The precipitate is then treated according to the process described in Example 1 (iii) (H) to

  
  <EMI ID = 298.1>

  
question.

Example 4

  
  <EMI ID = 299.1>

  
stand overnight at room temperature, a large amount of ethyl ether is added to the mixture, which

  
  <EMI ID = 300.1>

  
which is washed several times with ethyl ether to obtain a thick syrupy layer.

  
(ii) The syrupy material thus obtained is dissolved in

  
  <EMI ID = 301.1>

  
a 200 ml column of Chitosan. The column is eluted

  
with water-methanol (3: 1) and the eluate collected in fractions. Ninhydrin positive fractions are combined

  
  <EMI ID = 302.1>

  
  <EMI ID = 303.1>

  
colorless identical to that obtained in Example 1 (iii) (B).

Example 5

  
  <EMI ID = 304.1>

  
  <EMI ID = 305.1>

  
the reaction solution obtained reduces us pressure.

  
  <EMI ID = 306.1> mentions.

Example 6

  
  <EMI ID = 307.1>

  
(free base) in 15 ml of an oau-methanol mixture (1: 7) to which 1.5 g (6.8 moles) of acetate are then added

  
  <EMI ID = 308.1>

  
oxycarbonyloxyauccinimide in 7 ml of tetrahydrofuran. The roposor mixture is left at room temperature overnight and the solution is concentrated under reduced pressure.

  
  <EMI ID = 309.1>

  
a column of 200 ml of Chitosan and the effluent leaving the column is then treated as in Example 4

  
  <EMI ID = 310.1>

  
mentioned compound,

Example 7

  
Preparation of 3,6'-di-N- benzyloxycarbonylkanamycin A

  
500 mg (1.03 mole) of ka are suspended

  
  <EMI ID = 311.1>

  
and 272 mg (1.24 mole) of zinc acetate dihydrate is added to the suspension. The mixture is stirred at room temperature for 10 hours to form a practically transparent solution to which is then added, in small portions, for approximately 2 hours, 540 mg (2.27 moles) of N-benzyloxycarbonyloxyauccinimide. After allowing the resulting mixture to stand at room temperature overnight, add a large amount of ethyl ether, remove the separated oily material and wash it several times with ethyl ether to obtain a thick syrupy material.

  
A thin layer chromatography on silica gel of a sample taken from the syrupy material, using, as development solvent, chloro-

  
  <EMI ID = 312.1> benzyloxycarbonylkanamycin A (which is colored by spraying with sulfuric acid followed by heating};
- weak spot at Rf 0.28; <EMI ID = 313.1> bonylkanamycin A; and <EMI ID = 314.1>

  
Has not reacted.

  
There is practically no spot corresponding to tri-N-benzyloxycarbonylkanamycino A, spot

  
  <EMI ID = 315.1>

  
tion in a column of 100 ml of CM Sophadox C-25 resin (NH4 + form) previously wetted with dioxane water (1: 1). We then submit the column to the process

  
  <EMI ID = 316.1>

  
which removes the zinc cation and the desired product separates from the others to give 412 mg (51%) of the mentioned compound, as a colorless solid.

  
By way of comparison, the process mentioned above is repeated, but replacing the zinc acetate dihydrate with 308 mg (1.24 mole) of nickel (II) acetate tetrahydrate, with the result obtaining the 3.6'-

  
  <EMI ID = 317.1>

  
kanamycin A

  
500 mg (1.03 moles) of ka are suspended

  
  <EMI ID = 318.1>

  
1 g (4.55 moles) of zinc acetate dihydrate is added to the suspension. The mixture is stirred at room temperature until a homogeneous solution is formed, to which a solution of

  
  <EMI ID = 319.1> <EMI ID = 320.1>

  
at rest overnight at room temperature and then treated in the same way as in Example 1 (ii) and (iii)
(B) to give 722 mg (83%) of the mentioned compound, under <EMI ID = 321.1>

  
thylformamide 1: 2)

  
Elementary analysis

  
  <EMI ID = 322.1>

  
  <EMI ID = 323.1>

  
but by replacing the p-methoxycarbobenzoxy-pnitrophenyl ester with 220 mg (1.54 mole) of t-butoxycarbonylazide, the above compound is obtained in the form of so-

  
  <EMI ID = 324.1>

  
1 g (4.55 moles) of zinc acetate dihydrate are added

  
  <EMI ID = 325.1>

  
ambient until a homogeneous solution is formed to which a solution of 1.2 g (5.1 moles) of the p-nitrophenol ester of trifluoroacetic acid dissolved in 10 ml of dimethyl sulfoxide is then added. The mixture obtained is kept overnight at room temperature and then treated with ethyl ether as mentioned in Example 1 (ii). The syrupy material insoluble in ether is then treated as in Example 1 (iii) (A) to

  
  <EMI ID = 326.1>

  
1: 2). HERE) <EMI ID = 327.1>

  
(15 ml) and tetrahydrofuran (5 ml) and 1 g (4.55 moles) of zinc acetate dihydrate is added to the suspension,

  
  <EMI ID = 328.1>

  
in 3 ml of tetrahydrofuran. The reaction solution is brought to room temperature in one hour and then kept at this temperature for 3 hours. The reaction mixture is then treated with ethyl ether as in Example 1 (ii) and the syrupy material insoluble in ether is then treated by the method of Example

  
  <EMI ID = 329.1>

  
C 50.11; H 6.31; N 6.49%

  
  <EMI ID = 330.1>

Example 12

  
  <EMI ID = 331.1>

  
The reaction mixture obtained by the method of Example 8 but using 260 mg (2.6 moles) of anhy-

  
  <EMI ID = 332.1>

  
p-nitro-phenyl is treated as described in Example 1
(iii) (A). 525 mg (72%) of the men-

  
  <EMI ID = 333.1>

  
  <EMI ID = 334.1>

  
  <EMI ID = 335.1>

  
found C 44.20; H 7.07; N 7.85%

Example 13

  
  <EMI ID = 336.1>

  
  <EMI ID = 337.1>

  
  <EMI ID = 338.1>

  
  <EMI ID = 339.1>

  
  <EMI ID = 340.1>

  
  <EMI ID = 341.1>

  
tee as in Example 1 (il!) (H). The: posi-

  
  <EMI ID = 342.1>

  
carbon dioxide gas then concentrated to dryness. We

  
  <EMI ID = 343.1> <EMI ID = 344.1>

  
found C 46.31; H 5.98; N 6.31; S 6.55%

  
If the above reaction process is repeated but omitting the zinc acetate, practically no colorless solid is recovered.

Example 15

  
  <EMI ID = 345.1>

  
methylkanamycin A (free base) in 12 ml of dimethylaultoxide and 1 g (4.55 moles) of zinc acetate dihydrate is added to the suspension. The mixture is stirred at room temperature until a homogeneous solution is formed, to which a solution is then added over 30 minutes.

  
  <EMI ID = 346.1>

  
(1: 1). The mixture obtained is kept overnight at room temperature and then treated as in Example 1

  
  <EMI ID = 347.1>

  
(c 1, water-dimethylformamide, 1: 2).

  
The subsequent treatment of the mentioned compound according to a process similar to that described in the example

  
  <EMI ID = 348.1> <EMI ID = 349.1>

  
C 52.16; H 6.68; N 6.40 #

  
found C 51.99; H 6.75; N 6.20%

  
The subsequent processing of the mentioned compound is-

  
  <EMI ID = 350.1>

  
  <EMI ID = 351.1>

  
(80%) by repeating the process of Example 15, but

  
  <EMI ID = 352.1>

  
The subsequent treatment of the compound mentions according to a process similar to that indicated in the example.

  
  <EMI ID = 353.1>

  
born is obtained in the form of a colorless solid, with a

  
  <EMI ID = 354.1> <EMI ID = 355.1>

  
.

  
elementary hnalyae

  
  <EMI ID = 356.1>

  
to the suspension 1 g (4.55 moles) of zinc acetate dihydrate. The mixture is stirred at room temperature until

  
  <EMI ID = 357.1>

  
slowly add over an hour to the cooled solution,

  
  <EMI ID = 358.1>

  
The subsequent treatment of the mentioned compound according to a process similar to that described in the example

  
  <EMI ID = 359.1> <EMI ID = 360.1>

  
suspension. The mixture is stirred at room temperature for one hour to form a homogeneous solution to which a solution is then added for approximately one hour.

  
  <EMI ID = 361.1>

  
  <EMI ID = 362.1>

  
At room temperature, the reaction solution obtained is treated with a large volume of ethyl ether as mentioned in Example 1 (ii) to give a thick syrupy material.

  
The syrupy material is then treated in the same way as in Example 1 (iii) (A) but using

  
  <EMI ID = 363.1>

  
C 54.81; H 6.50; N 6.95%

  
found C 54.77; H 6.71; N 6.88%

  
The subsequent treatment of the compound mentioned according to a process similar to that described in Example 31 gives

  
  <EMI ID = 364.1>

  
Starting from 480 mg (1.03 mole) of 4'-deaoxykanamycin B, free base, (Cf "Bulletin of the Chemical

  
  <EMI ID = 365.1> holds this compound to form a colorless solid with a

  
  <EMI ID = 366.1>

  
followed by another agitation. A solution of 1.1 g (4.4 moles) of N-benzyloxycarbonyloxy- is slowly added over about an hour to the resulting solution.

  
  <EMI ID = 367.1>

  
the mixture stand overnight at room temperature. A large volume of ethyl ether is then mixed with the reaction solution in order to separate an oil deposit.

  
  <EMI ID = 368.1>

  
give a thick syrupy substance.

  
This syrupy material is washed several times with water, the N-acetylated zinc complex being destroyed by water and the released zinc cation eliminated at the same time as the excess of zinc acetate initially existing. We

  
  <EMI ID = 369.1>

  
containing N-acetylated dibekacin. The solid is subjected to thin layer chromatography on silica gel using chloroformethanol-aqueous ammonia 18% (1: 1: 1, lower phase) as the developing solvent.

  
  <EMI ID = 370.1>

  
The subsequent treatment of the compound mentioned according to the process similar to that described in the example <EMI ID = 371.1>

  
methyldibekine

  
500 mg (1.07 moles) of 6'-N-methyldibekine (free base) and 1.2 g (5.45 moles) of acetate are dissolved

  
  <EMI ID = 372.1>

  
the reaction solution stand overnight at room temperature and it is then treated as indicated in Example 23, thus obtaining 910 mg of the compound mentioned practically pure.

  
Subsequent treatment of the mentioned compound according to a process similar to that described in the example

  
  <EMI ID = 373.1>

  
dibekacin.

Example 25

  
  <EMI ID = 374.1>

  
This compound is obtained, in the form of a colored solid, with a yield of 730 mg (79%) by following the processes described in Example 1 (i), (ii) and (iii) A,

  
  <EMI ID = 375.1>

  
  <EMI ID = 376.1>

  
Subsequent treatment of the compound according to a

  
  <EMI ID = 377.1>

  
500 mg (1.03 mole) of kanamycin A (free base) are suspended in 20 ml of dimethyl sulfoxide and 0.5 g (2.3 moles) of zinc acetate dihydrato are

  
  <EMI ID = 378.1>

  
ambient until a homogeneous solution is formed, to which 283 mg (1.13 mole) of N- are then added.

  
  <EMI ID = 379.1>

  
mixture obtained overnight at room temperature and then treated as in Example 1 (ii) and (iii) (I) to obtain 556 mg of the compound mentioned in the form of

  
  <EMI ID = 380.1>

  
free (Cf "Bulletin of the Chemical Society of Japan", vol. 50, p. 1580-1583 (1977 ", we joust to the solution

  
  <EMI ID = 381.1>

  
N-benzyloxycarbonyloxysuccinimide. After allowing the mixture to stand overnight at room temperature, the reaction solution obtained is treated with a large volume of ethyl ether, as mentioned in Example 1 (ii) to obtain a thick syrupy material.

  
The syrupy material is then treated as in Example 1 (iii) (B) but using water-dioxane

  
  <EMI ID = 382.1>

Example 29

  
  <EMI ID = 383.1>

  
The compound mentioned, we form a colorless solid, is obtained with a yield of 780 mg following the process of Example 28 but starting with 500 mg (1.12 m

  
  <EMI ID = 384.1>

  
etc ...)

  
  <EMI ID = 385.1>

  
hydroxybutyric. The mixture is allowed to stand at room temperature for 10 hours. The reaction solution obtained is concentrated in small volume, and the concentrate is taken up in 4 ml of water-dioxane (1: 1). A small amount of acetic acid is added to the solution to make it weakly acidic, then subjected to a

  
  <EMI ID = 386.1>

  
under atmospheric pressure for one hour in the presence of palladium black (to remove the benzyloxycarbonyl group). The reaction solution obtained is filtered and concentrated and the concentrate is made to think in a

  
  <EMI ID = 387.1>

  
to elution by gradients with concentrations of am- <EMI ID = 388.1>

  
holding the desired product are combined and concentrated to dryness to give 24 mg (yield 60%) of the compound mentioned, we form a monocarbonate whose physical properties "and antibacterial activity are identical to that of an authentic sample.

  
Example 32 (Reference)

  
  <EMI ID = 389.1>

  
nyl-dibékacine prepared as in Example 23, are dis-

  
  <EMI ID = 390.1>

  
add 13 mg (0.12 mole) of N-hydroxyauccinimide ester

  
  <EMI ID = 391.1>

  
stand at room temperature and then it is treated

  
  <EMI ID = 392.1>

  
  <EMI ID = 393.1>

  
the physical properties and antibacterial activity are identical to those of an authentic sample.

Example 33

  
  <EMI ID = 394.1>

  
  <EMI ID = 395.1>

  
ambient ture. After addition to the reaction mixture of a

  
  <EMI ID = 396.1>

  
reaction reaction in a large volume of ethyl ether and the deposited oily material obtained is washed well

  
  <EMI ID = 397.1>

  
This material is well dried to give 640 mg of the compound

  
  <EMI ID = 398.1>

  
  <EMI ID = 399.1>

  
with 6 mg of phenyl trifluoroacetate and the mixture is left

  
  <EMI ID = 400.1>

  
The reaction mixture is then treated as in Example 33, which gives 24.8 mg of the mentioned compound which has

  
  <EMI ID = 401.1>

Example 35

  
  <EMI ID = 402.1>

  
Kanamycin A in 0.3 ml of hexamethylphoephoric triamide is mixed with 7 mg of ethyl trifluoroacetate and the mixture obtained is left to stand overnight at room temperature. The reaction solution is mixed

  
  <EMI ID = 403.1>

  
in a large volume of ethyl ether. The oily material deposited is washed well with ethyl ether and the solid substance dried to give 11.7 mg (yield

  
  <EMI ID = 404.1>

  
racetate, solid.

  
  <EMI ID = 405.1>

  
  <EMI ID = 406.1>

  
cetyl kanamycin A

  
7 mg of ethyl trifluoroacetate are mixed with a

  
  <EMI ID = 407.1>

  
  <EMI ID = 408.1>

  
Lange obtained stand overnight at room temperature. The homogeneous reaction solution thus obtained

  
  <EMI ID = 409.1>

  
then poured into a large volume of ethyl ether. The ma-

  
  <EMI ID = 410.1>

  
  <EMI ID = 411.1>

  
  <EMI ID = 412.1> <EMI ID = 413.1>

  
cetylkanamycin A

  
  <EMI ID = 414.1>

  
A in 0.35 ml of sulfolane and the mixture is stirred overnight at room temperature. We then process the

  
  <EMI ID = 415.1>

  
cine A in 0.8 ml of tetrahydrofuran and the mixture is stirred for 2 days. The homogeneous reaction solution obtained is mixed with 15 mg of ethyl trifluoroacetate and 8 mg of anhydrous sodium carbonate, stirred overnight then left to stand for 2 days. The reaction solution obtained is concentrated and the concentrate washed

  
  <EMI ID = 416.1>

  
the lide is suspended in a small volume of tetrahydrofuran together with a small amount of trifluoroacetic acid. The mixture thus obtained is stirred, then ethyl ether is added. The solid which precipitates

  
  <EMI ID = 417.1>

  
  <EMI ID = 418.1>

  
  <EMI ID = 419.1> of ethyl in 0.1 mg of tôtrnhydrofuranno; the mixture obtained is maintained at room temperature laying one day.

  
  <EMI ID = 420.1>

  
0.1 ml of tetrahydrofuran, and the mixture is left at room temperature for one day. The reaction solution

  
  <EMI ID = 421.1>

  
gives 2.3 mg (yield 18%) of the product mentioned under

  
  <EMI ID = 422.1>

  
add to the suspension obtained 1 g (4.55 moles) of zinc acetate dihydrate. The mixture is stirred at room temperature until a homogeneous solution is formed, to which 370 mg (2.59 moles) of t-butoxycarbonylazide are then added. The mixture obtained is left overnight at room temperature and then treated as described in Example 1 (ii) and (iii) (B) to obtain 590 mg
(80%) of the mentioned compound forms us a colorless solid.

  
  <EMI ID = 423.1>

  
  <EMI ID = 424.1>

  
ambient temperature.

  
The reaction solution is then treated as described in Example 33 which gives 76.8 mg (Yield

  
  <EMI ID = 425.1>

  
0.4 ml of dimethyl sulfoxide is mixed with 18 mg of ethyl trifluoroacetate and the mixture kept overnight at room temperature. The reaction solution is then treated as in Example 33, which gives

  
  <EMI ID = 426.1>

  
  <EMI ID = 427.1>

  
  <EMI ID = 428.1>

  
  <EMI ID = 429.1>

  
Example 10) and 12 mg of triethylamine are mixed with

  
0.6 ml of dimethyl sulfoxide then with 35 mg of trifluoro-

  
  <EMI ID = 430.1>

  
  <EMI ID = 431.1>

  
tional is then treated as in Example 33, which gives 94.2 mg (yield 96%) of the compound mentioned,

  
  <EMI ID = 432.1>

  
dimethylformamido, 1: 2).

  
Elementary analysis

  
  <EMI ID = 433.1>

  
found C 35.09; H 3.99; N 6.07%

Example 44

  
Production of 3, 6 '-di-N-phenoxycarbonyl-3 "-N-t.rifluoracetylkenamycin A

  
  <EMI ID = 434.1>

  
dipped at 23 mg of trifluoroacetato of methyl and the mixture is then treated as in Example 33 which gives

  
  <EMI ID = 435.1>

  
  <EMI ID = 436.1>

  
of dimethyl tallow oxide is heated at 100 ° C. for 12 hours in a sealed tube to carry out the 3 "-N-formylation. The reaction solution is mixed with a small amount of formic acid and then poured into a large volume of ethyl ether and treated as in example 33, this

  
  <EMI ID = 437.1>

  
solid form with positive reaction to ninhydrin.

  
  <EMI ID = 438.1>

  
Elementary analysis <EMI ID = 439.1>

  
N-methylkanamycin A (see Example 15) and triethylamine (11 mg), 30 mg of ethyl trifluoroacetate and 0.7 ml of dimethyl sulfoxide is treated as in Example 33, this

  
  <EMI ID = 440.1>

  
mine (11 mg) in 0.4 ml of dimethyl sulfoxide is mixed with 21 mg of ethyl trifluoroacetate and the mixture kept overnight at room temperature. The reaction solution is then treated as in Example 33, which gives 64.8 mg (yield 97%) of the compound mentioned, under

  
  <EMI ID = 441.1>

  
1: 2).

  
Elementary analysis

  
  <EMI ID = 442.1>

  
  <EMI ID = 443.1>

  
(c 1, water-dimethylformamide 1: 2).

Example 49

  
  <EMI ID = 444.1>

  
  <EMI ID = 445.1>

  
mixture is maintained overnight at room temperature. The

  
  <EMI ID = 446.1>

  
dichloroacetic acid then treated as in the example

  
  <EMI ID = 447.1>

  
thylformamide 1: 2). D

  
Elementary analysis

  
  <EMI ID = 448.1>

  
found C 46.58; H 5.33; N 5.62; Cl 14.28%

Example 50

  
  <EMI ID = 449.1>

  
it is mixed with a small volume of trichloroacetic acid and then treated as in Example 33, which gives

  
  <EMI ID = 450.1>

Example 51

  
  <EMI ID = 451.1>

  
  <EMI ID = 452.1>

  
thyl and the mixture is kept overnight at room temperature. The reaction solution is then treated

  
  <EMI ID = 453.1>

  
(12 mg) and 30 mg of ethyl trifluoroacetate in 1 ml of dimethyl sulfoxide is treated as in Example 33,

  
  <EMI ID = 454.1>

  
water-dimethylformamide, 1: 2).

Example 53

  
  <EMI ID = 455.1>

  
  <EMI ID = 456.1>

  
  <EMI ID = 457.1>

  
1: 2).

Example 55

  
  <EMI ID = 458.1>

  
and the mixture is kept overnight at room temperature. The reaction solution is mixed with a small amount of dichloroacetic acid and then treated

  
  <EMI ID = 459.1>

  
mono-dichloroacetate of the mentioned compound, in the form

  
  <EMI ID = 460.1>

  
  <EMI ID = 461.1>

  
  <EMI ID = 462.1>

  
1: 2).

Example 57

  
  <EMI ID = 463.1>

  
bonyl-tobramycin (see example 20) and triethylamine <EMI ID = 464.1>

  
  <EMI ID = 465.1>

  
as in Example 48: 86 mg is obtained (yield

  
  <EMI ID = 466.1>

  
  <EMI ID = 467.1>

  
C 55.98; H 6.09; N 7.42% found C 55.50 H 6.22; N 7.28%

Example 58

  
  <EMI ID = 468.1>

  
triethylamine (12 mg) in 1.2 ml of dimethyl sulfoxide is mixed with 30 mg of ethyl trifluoroacetate and the mixture is treated as in Example 33 i is obtained

  
  <EMI ID = 469.1>

  
  <EMI ID = 470.1>

  
dimethylformamide, 1: 2).

Example 59

  
  <EMI ID = 471.1>

  
  <EMI ID = 472.1>

  
and the mixture is treated as in Example 33; 100 mg (98% yield) of the mentioned compound are obtained, under

  
  <EMI ID = 473.1>

  
1: 2).

  
Elementary analysis

  
  <EMI ID = 474.1> <EMI ID = 475.1>

L

  
dimethylformamide, 1: 2).

Example 62

  
  <EMI ID = 476.1>

  
chloracetyl-dibekacin

  
An 84 mg solution of 3.2 ', 6'-tri-N- is made up

  
  <EMI ID = 477.1>

  
  <EMI ID = 478.1>

  
mono-dichloracetate of the desired compound, in solid form.

  
  <EMI ID = 479.1>

  
bonyl-6'-N-methyl dibékacine (see example 24) in 1 ml of dimethyl sulfoxide is mixed with 30 mg of ethyl trifluoroacetate and the mixture is treated as in Example 33;

  
  <EMI ID = 480.1> <EMI ID = 481.1>

  
thylsulfoxydo is mixed with 30 mg of ethyl trifluoracetato and the mixture is treated as in Example 33

  
  <EMI ID = 482.1>

  
(c 1, water-dimethylformamide, 1: 2).

Example 66

  
  <EMI ID = 483.1>

  
thylformamide, 1: 2).

Example 67

  
  <EMI ID = 484.1> <EMI ID = 485.1>

  
mamido, 1: 2).

Example 68

  
  <EMI ID = 486.1>

  
dimcthylformamido, 1: 2).

Example 69

  
  <EMI ID = 487.1>

  
thylsulfoxide is kept overnight at room temperature. The reaction solution is then treated as in

  
  <EMI ID = 488.1>

  
  <EMI ID = 489.1>

  
mixed with 13 mg of N-formylimidazole and the mixture is kept at room temperature overnight. The reaction solution is mixed with a small amount of formic acid and then treated with ethyl ether as in

  
  <EMI ID = 490.1>

  
formate of the mentioned compound in solid form.

  
  <EMI ID = 491.1>

  
thyl tallow oxide is. stirred, under ice cooling, laying 3 hours, then allowed to rest at room temperature overnight. The reaction solution is made alkaline by mixing with 0.3 ml of ammonia-

  
  <EMI ID = 492.1>

  
ambient. The resulting reaction mixture is treated with ethyl ether to obtain a syrup insoluble in ethor. The syrup is taken up in water and then passed through a column of CM-Sephadex C-25 (NH form).

  
  <EMI ID = 493.1>

  
column of resin is developed with 0.05 N ammonia. The fractions containing the desired product eliminated by elution are combined and concentrated to dryness. The concentrate is taken up in water, the aqueous solution neutralized with acetic acid and then again concentrated to

  
  <EMI ID = 494.1>

  
found C 45.22; H 7.20; # 8, it%

Example 72

  
  <EMI ID = 495.1>

  
xyde (50 ml) and tetrahydrofuran (20 ml) is mixed

  
  <EMI ID = 496.1>

  
and the mixture obtained is stirred at room temperature until the reaction mixture forms a homogeneous solution. It takes about 4-5 hours for kanamycin A to

  
  <EMI ID = 497.1>

  
zinc namycin A-cation. The solution obtained is

  
  <EMI ID = 498.1>

  
(9.5 millimoles) of N-benzyloxycarbonyloxysuccinimide in 40 ml of a mixed tetrahydrofuran-dimethylsulfoxide solvent (1: 1 by volume). The reaction solution is then kept for 4 hours at room temperature.

  
  <EMI ID = 499.1>

  
Kanamycin A is subjected to benzyloxycarbonylation. The reaction solution obtained is subjected to thin layer chromatography on silica gel using, as development solvent, the lower layer of chloroform-methanol-ammonia at 28% (1: 1: 1 by volume); it is then observed that the silica gel plate shows a main spot at Rf 0.23 and two or three slightly perceptible minor spots are placed above the main spot and are attributable to other roua-produced.

  
(ii) The reaction solution containing

  
  <EMI ID = 500.1>

  
ethyl ether; the oily precipitate is then washed

  
  <EMI ID = 501.1>

  
of a syrupy product containing the N-benzyloxycarbonylated complex.

  
(iii) The elimination of the zinc cation from the syrupy complex product is carried out as described below using a weakly acidic cation exchange resin containing carbon functions.

  
  <EMI ID = 502.1>

  
  <EMI ID = 503.1>

  
(form il +) with water-dioxane (2: 1 by volume). We fill a column with this resin and then we pass <EMI ID = 504.1>

  
positive for ninhydrin, first pass, then the fractions containing zinc acetate, positive for coloring with diphenylcarbazide, are collected. The first fractions containing the desired product are combined and concentrated to dryness; the concentrate is washed with ether

  
  <EMI ID = 505.1>

  
Elementary analysis

  
  <EMI ID = 506.1> zyloxycarbonyl-3 "-N-trifluoracetylkanamycin A

  
The product obtained in the above process (a) is

  
  <EMI ID = 507.1>

  
1.5 mol equivalent of triethylamine which gives the compound mentioned.

  
c) Preparation of l-N (L-4-amino-2-hydroxybutyryl) kanamycin A

  
  <EMI ID = 508.1>

  
held in the above process (b) in 1.5 ml of autetrahydrofuran (1: 1 by volume) is mixed with 7 mg of anhydrous sodium carbonate and then 23 mg of Nhydroxysuccinimide ester of L- acid are added thereto 4-benzyloxy-carbonylamino-2-hydroxybutyric, and the mixture is maintained 10

  
  <EMI ID = 509.1>

  
The reaction solution thus obtained is concentrated in small volume and mixed with water, giving a solid precipitate. The precipitate is taken up in 3 ml of 2N ammonia-tetrahydrofuran (5: 3 by volume) and the solution is kept overnight at room temperature to

  
  <EMI ID = 510.1>

  
xanne (1: 1) and the solution made weakly acidic by adding a very small amount of acetic acid and subjected to catalytic hydrogenolysis with hydrogen

  
  <EMI ID = 511.1>

  
black palladium catalyst for removing benzyloxycarbonyl groups. The reaction solution obtained is filtered and concentrated, and the concentrate is passed through a column of CM-Sephadex C-25 (form

  
  <EMI ID = 512.1>

  
is then developed by gradients with an ammonia concentration ranging from 0 to 0.5 N. The fractions containing the desired product are combined and concentrated to dryness to give 36 mg (yield 89%) of the monocarbonate of the mentioned compound. The physicochemical properties and anti-bacterial activities of this product were found to be perfectly identical to those of an authentic sample.

Example 73

  
  <EMI ID = 513.1>

  
Kanamycin A (free base) in 12 ml of dimethyl sulfoxide is mixed with 1 g (4.55 millimoles) of zinc acetate dihydrate, and the mixture obtained is stirred until a homogeneous solution is formed. We add to this solution

  
  <EMI ID = 514.1>

  
bonyloxysuccinimide in 5 ml of dimethylaulfoxide-tetra .. hydrofuran (1: 1 by volume) and the reaction solution is kept overnight at room temperature. The reaction solution is then treated as in example-

  
  <EMI ID = 515.1> <EMI ID = 516.1>

  
The product obtained in step a) is treated as in Example 47.

  
c) Preparation of l-N (L-4-amino-2-hydroxybutyryl) - <EMI ID = 517.1>

  
mycine A obtained in step b) above in 1.5 ml

  
  <EMI ID = 518.1>

  
6 mg of anhydrous carbonate of sodiur.i, then 20 mg of N-hydroxyauccinimide ester of L-4-benzyloxy acid are added thereto.

  
  <EMI ID = 519.1>

  
naked 8 hours at room temperature. The reaction solution is concentrated to a small volume and mixed with water, which produces a solid precipitate.

  
  <EMI ID = 520.1>

  
hydrofuran (1: 1 by volume) and the mixture is kept overnight at room temperature to effect the elimination of the 3 "-N-trifluoroacetyl group.

  
The reaction solution is concentrated to dryness to give a solid residue, and this residue is mixed with

  
  <EMI ID = 521.1>

  
weakly acidic due to the addition of a small amount of acetic acid and subjected to hydrogenolysis with hydrogen at atmospheric pressure for one hour on a black palladium catalyst to effect the elimination of the benzyloxycarbonyl groups. Then we treat the so-

  
  <EMI ID = 522.1>

  
72 (c) which gives 30 mg (87% yield) of the monocarbonate <EMI ID = 523.1>

  
lumo and we mix with water which gives a solid precipitate.

  
  <EMI ID = 524.1>

  
mix at room temperature overnight. The reaction solution is concentrated to dryness to give a solid residue. Lo residue is mixed with 6 ml

  
  <EMI ID = 525.1>

  
acid by addition of a small amount of acetic acid and then subjected to hydrogenolysis with hydrogen at atmospheric pressure for 1.5 hours on a black palladium catalyst. The reaction solution is then treated as in Example 72 (c) which gives
42 mg (yield 91 / *) of the monocarbonate of the compound men-

  
  <EMI ID = 526.1> tobramycin.

  
  <EMI ID = 527.1>

  
mycine (free base) in 12 ml of dimethylsulfoxydo is mixed with 1 g (4.55 millimolos) of zinc acetate dihydrate and the mixture is stirred for one hour. It is added dropwise, in about an hour, to the solution <EMI ID = 528.1>

  
the reaction mixture stand overnight at room temperature. The reaction solution obtained is treated with a large volume of ethyl ether as in the ex-

  
  <EMI ID = 529.1>

  
The syrupy complex product is then treated as in Example 72 (a) (iii) but the oau-dioxanno ratio (2: 1)

  
  <EMI ID = 530.1>

  
c 54, ni; H 6.50; N 6.95% found C 54.77; H 6.71; N 6.88% <EMI ID = 531.1>

  
The product obtained in n) is treated as in Example 57 to give the product mentioned.

  
  <EMI ID = 532.1>
12 mg of anhydrous sodium carbonate, then 40 mg of the N-hydroxysuccinimido ester of (L) -4-bonzyloxycarbonyl-amino-2-hydroxybutyriquo acid are added thereto. The mixture is kept at room temperature laying for 10 hours. The so-

  
  <EMI ID = 533.1>

  
  <EMI ID = 534.1>

  
solid precipitate.

  
The solid is suspended in 2 ml of a solution.

  
  <EMI ID = 535.1>

  
board is vigorously stirred at 60 ° C for 3 hours, then filtered to give a solid residue containing the

  
  <EMI ID = 536.1>

  
oau-dioxanno (1: 3), the solution is round, slightly acidic by the addition of a very small amount of acetic acid.

  
  <EMI ID = 537.1>

  
dium. The reaction solution is then treated as in Example 72 (c) and it is passed through the column

  
  <EMI ID = 538.1>

  
with ammonia concentrations ranging from 0 to 1 N.

  
  <EMI ID = 539.1>

  
This product coincides well with the authentic product.

Example 76

  
  <EMI ID = 540.1>

  
are mixed with stirring to 15 ml of dimethyl sulfoxide.

  
The solution is mixed with 1.4 g (6.4 millimoles) of zinc acetate dihydrate with stirring. A solution is added dropwise to the solution over about an hour.

  
  <EMI ID = 541.1>

  
the mixture stand overnight at room temperature. The reaction solution is then mixed with a large volume of ethyl ether to give an oily deposit containing mainly the compound mentioned, and dimethyl-

  
  <EMI ID = 542.1>

  
liquid and then washed with ethyl ether to give a thick syrupy product.

  
  <EMI ID = 543.1>

  
Thanks to this treatment, the initial zinc acetate oxcess is eliminated and the N-bonzyloxycarbonylated zinc complex is destroyed; 1.1 g of an insoluble solid residue are obtained <EMI ID = 544.1>

  
If pourtnn &#65533; the solid product is washed with a solu

  
  <EMI ID = 545.1>

  
The product obtained was treated a) above as in Example 59 to obtain the compound mentioned.

  
  <EMI ID = 546.1>

  
(1: 3) is mixed with 18 mg of anhydrous sodium carbonate, then 60 mg of N-hydroxysuccinimide ester is added

  
  <EMI ID = 547.1>

  
and the mixture is kept at room temperature for 9 hours. The reaction solution is concentrated at low volume and mixed with water to allow it to settle.

  
a solid precipitate.

  
The precipitate is mixed with 12 ml of 4 N-tetrahydrofuran ammonia (1: 3) and the mixture is left to stand overnight at room temperature. The reaction solution

  
  <EMI ID = 548.1>

  
solid sidu. The solid obtained is dissolved in 12 ml of water-dioxane (1: 3), the solution made weakly acid by the addition of a very small amount of acetic acid and subjected to hydrogenolysis at atmospheric pressure for 1.5 hours on black palladium. The reaction solution is then treated as in Example 75 (c) which

  
  <EMI ID = 549.1> <EMI ID = 550.1>

  
It can be observed that the physicochemical properties and the antibacterial activities of this product coincide with those of an authentic sample (Journal of Antibiotics, vol 26, p. 412 (1973).

Example 77

  
Synthesis of 1-N (DL-3-amino-2-hydroxypropionyl) -dibékacine, it is at. say 1-N-DL-isoseryldibekine

  
  <EMI ID = 551.1>

  
of Example 59 in 5 ml of water-tetrahydrofuran (1: 3) is mixed with 16 mg of anhydrous sodium carbonate, then 51 mg of N-hydroxy-succinimide ester of a- are added thereto

  
  <EMI ID = 552.1>

  
(or DL-3-benzyloxycarbonylisoserine). The mixture is kept at room temperature for 10 hours. The reaction solution is then treated as in the example

  
  <EMI ID = 553.1>

  
water).

  
The physicochemical properties and antibacterial activities of this product were found to be identical to that of an authentic sample.

Example 78

  
  <EMI ID = 554.1> xycarbonyldibékacine.

  
500 mg (11.1 mole) of dibeka are suspended

  
  <EMI ID = 555.1>

  
stir the suspension to form a solution to which

  
  <EMI ID = 556.1>

  
zinc dihydrate. A solution of 1.17 g is added dropwise to the resulting solution, over about 30 minutes.
(3.86 moles) of p-methoxy-carbobenzoxy-p-nitrophenyl ester dissolved in 20 ml of dimethylsulfoxide and the mixture is kept overnight at room temperature. The aolu-tion obtained is then treated as described in example-

  
  <EMI ID = 557.1>

  
  <EMI ID = 558.1>

  
b) Preparation of the trifluoroacetate of 3,2 ', 6'-triN-p-methoxybenzyloxycarbonyl-3 "-N-trifluoracétyldibékacine.

  
  <EMI ID = 559.1>

  
mamide 1: 2).

  
c) Preparation of l-N (L-4-amino-2-hydroxybutyryl) dibékacine

  
  <EMI ID = 560.1>

  
  <EMI ID = 561.1>

  
  <EMI ID = 562.1>

  
amino-2-hydroxybutyric, and the mixture is kept for 8 hours at room temperature. The reaction solution is concentrated to a small volume and mixed with water, leading to the deposition of a solid precipitate.

  
A solution of HCl in aqueous methanol (1: 3, 6 ml) is added to the solid and the mixture is heated at 60 ° C for 4 hours to remove the p-methoxybenzyloxy carhonyl group. Concentrate the solution at low volume, then

  
5N ammonia is added thereto until the pH of the solution is 10. The solution is kept overnight at room temperature and concentrated to obtain a residue. The residue is dissolved in water, the solution loaded into a column of CM-Sephadex C-25 (form

  
  <EMI ID = 563.1>

  
by gradients with ammonia concentrations ranging <EMI ID = 564.1>

  
  <EMI ID = 565.1>

  
bonate).

  

  <EMI ID = 566.1>
 

  
  <EMI ID = 567.1>

  
selectively acylated, of an aminoglycoside antibiotic, said antibiotic comprising a deoxystreptamine fraction having a 3-aminoglycosyl or 3-alkylaminoglycoayl group linked to its 6-hydroxy group and the N-protected derivative, selectively acylated, having certain of its amino groups selectively protected by an acyl group, characterized

  
in that it consists of
(a) reacting an acylation reagent having an acyl group to be introduced as an amino-protecting group with a <EMI ID = 568.1>

  
which was formed by reacting the aminoglycoside antibiotic with a zinc salt in an inert organic solvent, to produce a complex of zinc cations with

  
the selectively N-acylated derivative of the aminoglycosidic antibiotic, derivative in which the initially uncomplexed amino groups are acylated, and
(b) reacting the zinc cation complex with the selectively N-acylated derivative of the aminoglycoside antibiotic with a reagent which removes the zinc cations from said complex, to produce the N-protected derivative, selected <EMI ID = 569.1>


    

Claims (1)

<EMI ID = 570.1> 3.- Method according to claim 1, characterized in that the aminoglycoside antibiotic is kanamy- <EMI ID = 571.1> <EMI ID = 572.1> 4.- Method according to claim 1, characterized in that the formation of the antibiotic aminoglycosidiquo-cation do zinc complex is carried out by reacting acetate or zinc chloride to an amount <EMI ID = 573.1> nol, aqueous methanol, ethanol and aqueous ethanol, with or without the addition of sodium acetate. 5.- Method according to claim 1, characterized in that the acyl group of the acylation reagent used is an alkanoyl group, an aroylc group, an alkoxycarbonyl group, an aralkyloxycarbonyl group, an aryloxycarbonylc group, an alkylsulfonyl group, an aralkylaulfonyl group or an arylsulfonyl group known as an amino protecting group. 6.- Method according to claim 1, characterized in that the acylation reagent is used in a molar amount equal to or slightly greater than the number of amino to acylor groups in the antibiotic aminoglycoside-zinc cation complex. 7.- Method according to claim 1, characterized in that the complex of zinc cations with the derivative <EMI ID = 574.1> is first separated from the acylation reaction mixture before being reacted with the zinc cation removal reagent it contains. 8.- Method according to claim 1, characterized in that "the complex of zinc cations with the selectively N-acylated derivative of the aminoglycoside antibiotic is separated from the mixture of the acylation reaction by extraction with an organic solvent, by evaporation of the organic solvent medium contained in the acylation reaction medium or by dilution of the acylation reaction medium with a diluting organic solvent, before being reacted with a reagent for removing zinc cations. 9. A method according to claim 1, characterized in that the zinc cation cation complex with the selectively N-acylated derivative of the antibiotic aminoglycosidiquo, <EMI ID = 575.1> organic polar, either anhydrous or aqueous, which leaves the reagent for the elimination of zinc cations. 10.- Method according to claim 9, characterized in that the polar organic solvent is, either do those in which the zinc salt is soluble, but the derivative <EMI ID = 576.1> either of those in which the zinc salt is insoluble, but the N-acylated derivative of the aminoglycoside antibiotic is soluble. 11.- Method according to claim 1, characterized in that the complex of zinc cations with the N -acylated derivative of the aminoglycosidic antibiotic, once separated, is again dissolved entirely in an organic solvent containing a proportion of after which the new solution thus obtained is subjected to a chromatographic treatment using a cation exchange resin, an anion exchange resin, a chelate exchange resin or a polymer insoluble in water and containing functional groups capable to combine with a metal, which acts as a reagent for the elimination of zinc cations. 12.- Method according to claim 1, characterized in that the acylation reaction mixture is passed directly through a column of a cation exchange resin, an anion exchange resin, an exchange resin of chelates or of a polymer insoluble in <EMI ID = 577.1> metals to achieve adsorption of the cation complex <EMI ID = 578.1> cosidiquo, then we develop it. column with an aqueous organic solvent containing or not a proportion of a- <EMI ID = 579.1> after which the fractions which contain the desired selectively N-acylated derivative of the aminoglycoside antibiotic are recovered without containing zinc cations. 13.- Method according to claim 1, characterized in that when the desired N-acylated derivative of the aminoglycosidic antibiotic is insoluble or substantially insoluble in water, the reaction mixture is immediately incorporated with water, so that said derivative is precipitated by separating from the zinc salt which remains dissolved in water. 14.- Method according to claim 1, characterized in that the reaction acylation mixture is treated with hydrogen sulfide, an alkali metal sulfide <EMI ID = 580.1> zinc cations in the form of zinc sulfide, or with ammonium hydroxide, which precipitates zinc cations in the form of zinc hydroxide. 15.- Process for the preparation of an N-acylated derivative, selectively protected, of an aminoglycoside antibiotic <EMI ID = 581.1> 1-amino group do the deoxystreptaminc fraction is not protected, but all the other amino groups of the aminoglycoside molecule are protected by identical or different acyl groups, characterized in that it consists of a single operation which is (a) reacting an alkanoic acid ester of formula (VIII): <EMI ID = 582.1> <EMI ID = 583.1> di- or trihalogenated alkyl containing from 1 to 6 do atoms <EMI ID = 584.1> carbon atoms, an aralkyloxy group, especially the bonzyloxy group or an aryloxy group, especially the phenyloxy group, or an N-formylimidazolo, as acylating agent, in an inert organic solvent, with an N-acylated derivative, partially protected, of the amino antibiotic <EMI ID = 585.1> <EMI ID = 586.1> while all other amino groups are protected by an acyl group as a non-protective friend group. for <EMI ID = 587.1> dideoxykanamycin C, gentamycin A, gentamy- <EMI ID = 588.1> somycin or netilmycino. 17.- Method according to claim 15, characterized in that the alkanoic acid ester of formula (VIII) <EMI ID = 589.1> thyle, butyl formate, benzyl formate, <EMI ID = 590.1> ethyl dichloroacetate, methyl trichloroacetate, phenyl trichloracetate, methyl trifluoroacetate, ethyl trifluoroacetate or phenyl trifluoroacetate. 18.- Method according to claim 15, characterized in that N-formylimidazole is used as acylating agent. 19.- Method according to claim 15, character- <EMI ID = 591.1> temperature between -30 * C and + 120'C for a period of 30 minutes to 24 hours, even 48 hours, in a soil- <EMI ID = 592.1> nitromethane, sulfolane, dimethylacetamide, chloroform, dichloromethane, methanol, ethanol, <EMI ID = 593.1> sulfuric ether, either anhydrous or aqueous. 20.- Improved process for preparing a <EMI ID = 594.1> characterized in that it consists of; (a) reacting zinc cations with the antibiotic <EMI ID = 595.1> (b) reacting an acylating reagent having an acyl group to be introduced as an amino-protecting group with the com- <EMI ID = 596.1> to produce a zinc cation complex with the selectively N-acylated derivative of the aminoglycoside antibiotic. derivative in which the amino groups initially <EMI ID = 597.1> (c) react the selectively N-acylc derivative complex <EMI ID = 598.1> in (b) with a reagent which eliminates the zinc cations of the said complex in order to obtain an N-acylated derivative, partially and selectively protected from the aminoglycosidic antibiotic, which is free from zinc cations and in which the <EMI ID = 599.1> are not protected, all the other amino groups of the aminoglycoaide molecule being protected by the acyl group, (d) reacting the partially and selectively protected N-acylated derivative obtained in (c) with an alkanolate acid ester of formula (VIII): <EMI ID = 600.1> wherein R is a hydrogen atom or a di- or trihalogenated alkyl group containing from 1 to 6 carbon atoms, and R is an alkyloxy group containing from 1 to 6 carbon atoms, an aralkyloxy group containing from 1 to 6 atoms carbon or an aryloxy group, or N-formylimidazole, as acylating agent, in an organic solvent <EMI ID = 601.1> <EMI ID = 602.1> in which m is equal to 1 or 2, or an equivalent reactive derivative of this acid, the amino group of which is protected or not, to acylate the 1-amino group of the derivative obtained in (d), and (f) removing the residual amino protective rumps from the 1-N-acylation product obtained in (e) by a traditional "deprotection" process. <EMI ID = 603.1> dique, substantially as described above.