CS268156B2 - Process for preparing 1,1-dioxydes of 6-aminoalkylpenicillanic acids - Google Patents

Abstract

The title compds. I and II [ R=H; n=1; Q=benzyl, o-, m-, phydroxybenzyl, penethyl, 2-, 3-, 4-picolyl; R1=H, ester! were prepd. Thus, 108.73g benzyl-6,6-dibromo-penicillanate in 600 ml THF (-78≰C) was reacted with 83.5 ml 2.9 M-methylmagnesium in ether soln., mixed with 27 g benzyloxycarboxy amido methyl acetate in 200 ml THF for 10 min., and treated with 14.52 ml acetic acid to give 113 g benzyl-6-α-bromo-6-β-(benzyloxycarbonyl-amino methyl) penicillanate.

Description

The present invention relates to a process for the preparation of 1,1-dloxides of 6L- and 6B- (aminomethyl and 1-aminoethyl) -phenolioic acids, their pharmaceutically acceptable, in vivo hydrolysable esters, their bis-methanediol esters or their mixed methandiol esters of these inhibitors of β-lactamase and erulbaotam (1,1-penioic acid dioxide) · which methandiol esters are also hydrolyzable in vivo · Although some of the above compounds themselves show antibacterial activity · their essential importance lies in their activity As β-lactamase inhibitors. They are therefore useful in combination with conventional β-lactam antibiotics (penicillins and oephalosporins) against microorganisms resistant or partially resistant to β-lactam antibiotics as they produce the enzyme β-lactamase. The invention also provides pharmaceutical compositions comprising a compound of the invention. [beta] -lactamase inhibiting agents, and the known [beta] -lactamase-type antibiotic, the bis-methandiol esters of the [beta] -lactamase inhibitors according to the invention, either and ampioillin or amoxicillin, pharmaceutical compositions containing these bis-esters, ? compositions and compounds useful as intermediates in the preparation of the above compounds;

Related compounds, such as Lactic Acid 1,1-dioxide and the corresponding esters readily hydrolyzable in vivo (Barth, U.S. Pat. No. 4,234,579), eulbactam bis-methanediol-eeter (Bigham, U.S. Pat. No. 4,309,347) · various 6-O- (hydroxymethyl) penicillanic acid 1,1-dioxides and the corresponding esters (Kellogg, U.S. Pat. No. 4,287,181) and 60- (aminomethyl) foams linic acid (McCombie, U.S. Pat. No. 4,237,051) have been described as O-lactamase inhibitors useful in combination with β-lactam antibiotics for the treatment of bacterial infections. Antibacterial active methandiol bs-esters of penicillins and penicillinic acid 1,1-dioxide have also been described (Bigham, U.S. Pat. No. 4,244,951 to Godtfredsen et al., U.S. Pat. No. 4,342,772).

British Patent Application 2,053,220, published Feb. 4, 1921, discloses broadly disclosed β-lactamase inhibitory compounds corresponding to the general formula

C

The present meanings of the symbols R _a , R _b and R _c define practically an infinite number of compounds. These meanings are a suitable choice of the symbols R b, R _b and R _c may comprise a simple one 1,1-dioxide 6 <6 * L- and 6- (aminoalkyl) a foaming liánové acid according to the present invention. However, no specific method for the preparation of these compounds is disclosed in the aforementioned British patent application, nor is there any mention or even claim that of an indeterminate number of compounds described herein, the aminomethyl and 1-aminoethyl derivatives of the present invention are preferred substances exhibiting particularly high activity as Β-lactamase inhibitors, whose efficacy We have found in them.

The invention describes a process for the preparation of compounds of general formula I * and II

(II

And which is hydrogen or methyl; and.

has a value of 2 and R 1 is a group or has a value of 1 and R ¹ is a hydrogen atom; an ester-forming moiety hydrolysable under physiological conditions;

U) wherein the hydrogen atom is a hydroxyl group, a C 2 -C 7 alkanoyloxy group, a C 2 -C 7 alkoxycarbonyloxy group, a benzoyloxy group or a benzoyloxy group mono-substituted with a C 1 -C 4 alkyl group, a C 1 alkoxy group to 4 carbon atoms or halogen and their pharmaceutically acceptable acid addition salts and pharmaceutically-acceptable kationiokýoh salts of such compounds in which R ¹ is hydrogen, is further characterized in that, in an inert solvent in the presence of a catalyst based on a noble metal hydrogenolyzed the compound of formula III

R ₆ C ₆ H ₅ CH ₂ OCNHCH * or IV

And wherein R znamená is -CH2- or and R ^ is benzyl, an ester forming an hydrolysis physiological condition; a dioxopenicilancyloxymethyl group, and wherein

H *

(B) is hydrogen, benzyloxycarbonyloxy, C2-C7 alkanoyloxy, C2-C7 alkoxycarbonyloxy, benzoyloxy or benzoyloxy mono substituted with C1-C4 alkyl, C1-C4 alkoxy or alkoxy and

CS 26θ 156 B2

Z is azido or benzyloxycarbonylamino, and the resulting compound of formula (I) or (II) is optionally converted to its pharmaceutically acceptable acid addition salt or, when R ¹ is hydrogen, to its pharmaceutically acceptable cationic salt.

It will be understood that when R is methyl, the formulas I * and II * represent two different diastereomers (epimers). Depending on the absolute configuration, the side chains of these epimeric pairs are referred to as 1R-aminoethyl and 1S-aminoethyl. In each of these cases, the side chain has the R configuration in one eplmer and the S configuration in the other eplmer. The same is true for the compounds of Formulas III and IV, V and VI, and VII and VIU below. These pairs of isomers can generally be separated by column chromatography in a series of reaction steps during their preparation, as described in detail below.

Pharmaceutically acceptable acid addition salts include, but are not limited to, salts with hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, citric acid, maleic acid, succinic acid, benzenesulfonic acid, p-toluenesulfonic acid, 2-naphthaleneulfonic acid and methanesulfonic acid. Pharmaceutically acceptable cationic salts include, but are not limited to, sodium, potassium and calcium salts, salts with N, N-dibenzylethylenedlamine, N-methylglucamine (meglumine), and diethanolamine.

References to esters hydrolyzable under physiological conditions refer to those esters which are often referred to as prodrugs. These esters are well known recently and as common in the field of penicillin chemistry as pharmaceutically acceptable salts. Such esters are generally used to increase oral absorption however, they are readily hydrolyzed in vivo to the original free acid. Preferred esters do not tend to hydrogenolyze under the conditions preferably used to prepare them (see below). Preferred ester-forming residues are:

A N - butyrolacton-4-yl radical, a -CHR ² OCOR ³ radical and a -CHR ² OCOOR ³ radical, of 3 wherein R represents a hydrogen atom or a methyl group and Iv represents an alkyl group of 1 1 to 6 carbon atoms.

Most preferred ester-forming residues are pivaloyloxymethyl and 1-ethoxycarbonyloxyethyl.

When n is 1 and R ^{1 of the} formula represents 1,1-dioxopenicilanoyloxymethyl oo

and the compounds of formulas I * and II * are methandiol diesters. These esters are also hydrolyzable under physiological conditions to give the parent acid of formula I * or II *. wherein d is 1 and R ^ is hydrogen, and penicillanic acid 1,1-dioxide. The latter also acts as an β-lactamase inhibitor. If n is 2 and R ¹ is -CH ₂ - then the bis-ester is also hydrolyzed under physiological conditions, this time to give two molecules of the parent acid from each bis-ester molecule.

As the 6-β obeoného compounds of formula I * 6 «^ - compounds ^of formula II, regardless stereochemil side chain if R is methyl, are potent inhibitoxy0 -laktamaey. For oral administration, it is preferred that Ж is other than hydrogen. When R ¹ represents an ester moiety forming an ester moiety hydrolyzable in vivo, preferred residues of this kind are defined above. Especially? valuable compound

Of this group is pivaloyloxymethyl-6X- (aminomethyl) penioilane-1,1-dioxide p-toluenesulfonate because it is readily prepared in a highly pure crystalline state and the resulting product can be used as a pharmaceutically acceptable mammal salt. Also readily formulated and especially preferred for oral administration are the 6L- (aminomethyl) derivatives of formula (II) wherein D is 1 and R ¹ is 1,1-dioxopeniollanoyloxymethyl or where D is 2 and R ¹ is -CHg-.

Both 6 β-compounds of formula I * »and 6Z-compounds of formula II *», regardless of the side chain stereochemistry when R is methyl, are potent inhibitors of β-lactamase ·

The compounds of the formulas I * and II * »in which R ¹ is as defined above, with the exception of the remainder of formula A, hereinafter referred to as compounds of formula I and II, respectively. II, are useful as inhibitors of the enzyme β-lactamase. In this way, the compounds increase the activity of .beta.-lactam antibiotics (penioillins and cephalosporins), particularly against those microorganisms which are resistant or partially resistant to .beta.-lactam antibiotics by producing enzymes (.beta.-lactamases) which break down or They partially break down the β-lactam antibiotic · The presence of the compounds according to the invention thus leads to an increase in the activity of the β-lactam antibiotic · The lactam antibiotics form one of the most well known and widely used classes of antibacterial agents. a (-lactam) ring not fused to a thiazolidine or dlhydro-1,3-thiazine ring. When the core contains a thiazolidine ring, such compounds are commonly referred to as penicillins, whereas compounds having a core containing a dlhydrothiazine ring are referred to as cephalosporins. While the compounds of the present invention exhibit general activity in enhancing the activity of β-lactam antibiotics, they are preferably used in combination with penicillin or cephalosporin established in clinical practice such as amoxioillin, ampioillin, azlooillin, bao-ampioillin, carbenloline, carbenicillin-indanyl , carbenioillin-phenyl, cefaclor, oefadroxil, cefotexime, cefsaxin, cefsaxine, cefamoxole, cefamoxole, cefamoxole, cefamoxole, cefamoxole, cefamoxole, cefamoxin, cefotoxime, cefoxin, , cephaloglycine, cephaloridine, cephalothin, cepapirin, cepradine, cyclacline, epicillin, hetaoillin, levopropyleilin, meoilinam, meslooillin, penicillin G | penicillin V, phenethioillin, plperacillin, pirbenlcilin, plvampline, sarmoxicillin, sarpicillin, suncillin, talampicillin and ticarcillin, as well as pharmaceutically acceptable salts thereof. Preferred are combinations of the compounds of the invention in ampioillin or derivatives thereof, in amoxloillin or derivatives thereof, and in particular in cefoperazone.

While the compounds of the invention may be administered separately from the β-lactam antibiotic, combination dosage forms are nevertheless preferred. The pharmaceutical compositions, whether for oral or parenteral administration, contain both the .beta.-lactamase inhibitor of formula I or II and the .beta.-lactam antibiotic in a weight ratio of from 1: 3 to 3: 1, the total amounts of active ingredients being so high. to be sufficient for the successful treatment of bacterial infections in a mammal after a single, or more commonly, multiple administration.

The invention also encompasses compounds of the above general formulas (I) and (II) wherein R ¹ is a radical of formula (A) as defined above, which compounds are as follows:

(Ш)

o (IV)

CS 26a 156 В2

wherein it represents a hydrogen atom or a methyl group and represents a radical of formula k

wherein hydrogen, hydroxyl, C 2 -C 7 alkanoyloxy, C 2 -C 7 alkoxycarbonyloxy, benzoyloxy or benzoyloxy monosubstituted C 1 -C 4 alkyl, C 1 -C 4 alkoxy or halo; pharmaceutically acceptable acid addition mono- and disalts, suitable acids being described above *

These bis-methanediol esters of formulas III and IV are antibacterially effective because they are hydrolyzed in vivo to the corresponding O-lactamase inhibitors of formulas I and II and to the corresponding ampicillin, amoxicillin or amoxicillin substituted on phenolic oxygen. It should further be noted that these phenol esters are also generally hydrolyzed in vivo to form amoxioillin. The aforementioned bis-methanediol esters are formulated into pharmaceutical compositions suitable for either parenteral or oral administration in a single or (more usually) multiple dose for the treatment of bacterial infections in mammals.

Preferred compounds of formulas III and IV contain hydrogen or hydroxyl group, most preferred compounds contain hydrogen as Y.

The invention also includes intermediates of the following formulas (V),

VI, VII and VIII

R

CH.

(IN)

R

o (VI)

(VII)

in which

A represents a group

CS 2бе 156 B2

R

(VIII)

sulfur atom,

R represents a hydrogen atom or a methyl group;

Y * represents benzyloxycarbonylamino, amino, azido or trifluoromethanesulfonyloxy groups.

The compounds of the formulas I and II according to the invention are generally prepared from benzyl 6,6-dibromopeniltilate or from benzyl 6O-iodopenicilaeate.

A preferred process, in particular for the preparation of 6 β- (aminomethyl) derivatives, involves converting the dibromo derivative into a mixture of epimeric mono-Grignard reagents as a first reaction step. This conversion is expediently carried out by an exchange reaction using virtually one equivalent of methylmagnesium bromide in an ether solvent (ether, tetrahydrofuran, dimethoxyethane) at a low temperature (-50 to -100 ° C), conveniently at -78 ° C, which is the cooling bath temperature dry ice-acetone * · After a short reaction time (5 to 30 minutes) at such reduced temperature, the mono-Grignard reagents are contacted with virtually 0 _t 5 molar equivalents benzyloxykarboxamidomethyl-aoetátu which is usually diluted with the same etherlckým solvent and added к cold Grignard reagent at a rate to maintain a low reaction temperature. The reaction time does not play a decisive role. It is usually sufficient to operate from 0.5 to 2 hours at -50 to -100 ° C to complete the reaction. A mixture of epimers of the above formulas VII and VIII, wherein Y 'is a benzyloxycarbonylamino group, is readily obtained by the addition of acetic acid, concentration of the mixture and chromatography. This mixture of epimers can be used directly in the next step or optionally separated by further silica gel column chromatography.

The subsequent reaction step consists in the reductive cleavage of the bromine atom, which is usually carried out by an excess of tri-n-butyltin hydride, optionally in the presence of a small amount (less than 0.1 mol equivalents) of the free radical initiator, such as In the following, the term "inert solvent" refers to a solvent which does not react in the starting materials, reagents, intermediates or products in such a way as to cause a significant reduction in the yield of the desired product. In the present case, hydrocarbon solvents such as benzene or toluene are suitable. The reaction is to be carried out at an elevated temperature (60-100 ° C) to allow the reaction to proceed within a reasonable time, but not at an elevated temperature so as to cause undesirable thermal degradation. When this reaction step is carried out with a mixture of epimeric precursors, the 6β-epimer of formula V, where A is sulfur, is generally crystallized, and the 6β-epimer (VI, A «S) is optionally isolated from the mother liquors by evaporation and chromatography.

A second preferred process, in particular for the preparation of 6 &apos; - (aminomethyl) derivatives, consists in reacting a cold ethereal solution of a Grignard reagent prepared from benzyl-6-iodopenoicilate with benzyloxycarboxamidomethyl acetate under the conditions described above. It can be separated by column chromatography, but preferably this mixture is oxidized to 1,1-dioxides, which are then subjected to epimerization to Cg, as discussed in detail below to give pure epi7.

EN 156 B2 26θ burrow general formula VI wherein A represents a group S0 _second

For the production of the 1C and 16-oxides of the aforementioned formulas V and VI, in which A represents S group O or O, the abovementioned sulphides of the two or more of the formulas V and VI in which A represents sulfur are practically oxidized. mole equivalent of perkyeeline, suitably m-chloroperbenzoic acid, in an inert solvent such as ethyl acetate at a temperature of 0 to 50 ° C. When benzyl 6β- (benzyloxycarbonylaminomethyl) penicillanate (V, RH, A-S) is oxidized, the resulting d-oxide (V, R-H, A 0) is isolated by crystallization while the β-oxide (VI, R-H,

It is isolated from the mother liquors by evaporation. If desired, other standard reagents for preparing en1f oxides can also be used.

Optionally, the 10-benzyl-60- (benzyloxycarbonylaminomethyl) penicillanate (V, A-S-ag-O) 10 can be rearranged to the corresponding 6 (ε-epimer (VI, AS-β-O) by acting on the β-oxide. with an equivalent amount of 1,5-diazabioyklo [4,3.0] non-5-ane at 0 to 50 ° C, conveniently at 25 ° C, when the reaction is completed in a very short time (3 to 15 minutes). way six-dioxide (V, A _"2 S0) Transfer to GcG- corresponding epimer (VI, A - S0 _2). '

Oxidation of the sulfides of the formulas V and VI, where A is sulfur, or further oxidation of the above-mentioned sulfoxides, using an excess of peracid (otherwise under the conditions generally described above for the preparation of mono-oxides) ее yields the corresponding eulfones (1,1-dioxides) of the general in а formula VI in which A represents a group S0 _second If desired, other sulfone reagents such as potassium permanganate may be used.

Hydrogenolysis of the resulting benzyl-6 (C 1 or 4) - (benzyloxycarbonylaminoalkyl) -penicillanates (V and VI, A - SO ₂ ) to give the corresponding 6- (c * or 6) - (aminomethyl or 1-amino ethyl) -penicillanic acids (I and II) (R ¹ * H). Hydrogenolysis is carried out by methods well known in penioillin chemistry. The substrate is treated with hydrogen in an inert solvent in the presence of a noble metal catalyst such as palladium, platinum or rhodium, optionally in the form of an oxide or salt, or supported on a support such as activated carbon, alkali metal carbonate or alumina. The temperature is not critical and may range, for example, from 0 to 50 ° C, but is preferably operated at a temperature of 25 ° C or less to minimize thermal degradation. The pressure may be within a wide range (from subatmospheric pressure to 10 MPa), but is preferably operated at a pressure of from 0.1 to 0.7 MPa. As an inert solvent, a relatively low boiling solvent is preferably used which can be easily removed by concentrating the reaction mixture in vacuo. A particularly suitable solvent for this purpose is aqueous tetrahydrofuran. A preferred catalyst is palladium on carbon.

In order to prepare an in vivo hydrolysable ester, i.e. a compound of formula I or II in which R ^ represents an ester-forming moiety hydrolyzable under physiological conditions, the amino group of 6- (aroinoalkyl) 1,1-dioxide is first protected in a manner known per se using a benzyloxycarbonyl group. penicillanic acid. For example, benzyl chloroformate is slowly added to the amine in an inert solvent such as aqueous acetone or aqueous tetrahydrofuran, maintaining a pH of 8.0 at 0 to 35 ° C, preferably 0 to 20 ° C. In this way, compounds of the formulas (IX) and (X) below are those in which they are hydrogen. Alternatively, these compounds are obtained by the partial hydrogenolysis of compounds of formulas V а VI in which A represents a group S0 _2> In addition to limiting the consumption of hydrogen, the other reaction conditions employed as described above.

(X) (X)

CS 268 156 B2

These intermediates ее then convert to the desired esters corresponding to formulas IX and X in which R R represents an in vivo hydrolysable ester moiety by known methods conventional in penicillin chemistry (see, for example, U.S. Patent Nos. 4,234,579 and 4,287,181, and Preferred eether-forming moieties within the meaning of the symbol have been defined above, preferred methods for preparing such esters are described in detail in the examples below and in European Patent Publication No. 40,494.

The protected esters of formulas IX and X are converted to the desired esters of formulas I and II while maintaining the ether-forming residue by hydrogenolysis, preferably in the presence of a weakly acidic buffer consisting of equimolar amounts of a weak base amine such as pyridine and a strong acid such as a mineral acid. hydrochloric acid, nitric acid or sulfuric acid) or, preferably, sulfonic acids (such as methanesulfonic acid, benzenesulfonic acid or p-toluenesulfonic acid), otherwise using the methods described above, however, care must be taken to minimize exposure of the substrate to such conditions. (for example, the presence of water or lower alcohols, high acidity or alkali) which would lead to the hydrolysis of the ester or non-lactam moieties. It is preferable to isolate the ester directly from the reaction mixture in the form of an β acid addition salt where the acid is the same acid. A particularly preferred buffer is pyridine / p-toluenesulfonate, in which case the product is usually isolated as a salt with p-toluenesulfonic acid.

The above-mentioned pharmaceutically acceptable acid addition salts of a compound of the invention are readily prepared by standard procedures. For example, one equivalent of an acid is mixed with the corresponding compound in the form of a free amine, in an organic or aqueous organic solvent. The salt is isolated by concentrating and / or adding aero from the desert. In some cases, the salt is isolated directly from the reaction mixture without isolating the corresponding free amine.

The above-defined pharmaceutically acceptable cationic salts of those compounds of the invention that contain a free carboxyl group are also readily prepared by standard methods. For example, an equivalent amount of the respective hydroxide, carbonate, bicarbonate or amine is emitted by carboxylic acid in an organic or aqueous solvent, at a lower temperature (e.g. 0-5 ° C), the base is added slowly and the reaction mixture is vigorously stirred. The salt is isolated by concentration or addition of a non-solvent. In some cases, the salt is isolated directly from the reaction mixture without isolating the corresponding free acid.

Bis-methanediol esters of formulas III and IV as well as compounds of formulas I and II wherein n is 1 and R 1 is 1,1-dioxopenicilanoyloxymethyl, or wherein n is 2 and R 2 is also prepared from of amino-protected lineric acids corresponding to the formulas IX and X in which R ¹ represents a hydrogen atom. According to one alternative, the latter are first converted to the corresponding chloiraethylesters, which is preferably carried out by first converting the acid to its tetrabutylammonium salt, which is then reacted in an excess of chloromethyl iodide at a temperature of 0 to 50 ° C, preferably at a temperature of 25 ° C or less.

While the chloromethyl ester can be used directly in the next reaction step, it is nevertheless preferred to convert it first to the corresponding iodomethyl ester. This conversion is particularly advantageous in that the chloromethyl ester is not reacted with sodium iodide in acetone at 0 to 50 ° C until the reaction is complete.

CS 268 156 82

The iodomethyl ester 0 formed is then formed in an inert solvent at temperatures. 0 to 50 ° C is reacted with a salt of 1,1-dioxide of a penioilanoic acid, a salt of the same penioilanoic acid of the general formula XX or X, wherein R 6 represents a hydrogen protected, amino-protected, azidolefin p- (D-2-) salt phenyl-2-azidoacetamido) peniclian acid or with a salt of an ampicillin or amoxilcillin derivative of the general formula XI

O

I c ₂ c ₆ e ₅ (H) wherein

Y represents a hydrogen atom, a benzyloxycarbonyloxy group, an alkoxycarbonyloxy group having 2 to carbon atoms, a benzoyloxy group or a benzoyloxy group mono-substituted with a C1-C4 alkyl group, a C1-C4 alkoxy group or a halogen atom (fluorine, chlorine or bromine).

A preferred salt is the tetrabutylammonium salt, which reacts very rapidly with the iodomethyleater, thereby minimizing degradation of the molecule.

In accordance with the second alternative, the bis-methanediol-ethers of formulas III and IV, as well as esters of formulas I and II, wherein n is 1 and R ¹ is 1,1-dloxopenicilanoyloxyethyl, prepare the reaction of the above protected penicillanic acid salt protected. at an amino group corresponding to the general formula II or X in which R @ ¹ represents a hydrogen atom with a halomethyl ester (preferably iodomethyl ester) of penicillanic acid 1,1-dioxide, azidocillin or an ampicillin or amoxilcillin derivative of the above general formula XI.

In both of these alternatives, the resulting protected methanediol diester is then converted to the desired final product of formula (I) or (II).

1,1-dioxopenicilanoyloxymethyl or wherein n is 2 and R R is -CH ^-J, III or IV by hydro-genolysis using the methods discussed in more detail above, again minimizing the time the substrate is exposed to the conditions, which could lead to the cleavage of sensitive ester bonds of methanediol disetters. The pharmaceutically acceptable mono- and di-acid addition salts of these methanediol esters are prepared by the above-described methods using one or two equivalents of acid, as appropriate for the particular case.

A further preferred process leading to intermediates of formulas VII and VIII wherein Y * is a benzyl oxycarbonylamino group, especially when R is a methyl group, consists in the preparation thereof from the corresponding known benzyl-6 (E - (hydroxymethyl or 1-hydroxyethyl) 6β-bromopenicllanate and benzyl 6 (3- (hydroxymethyl or 1-hydroxyethyl) -6-

CS 268 156 B2 Bromoponyl aclanate (also prepared from benzyl 6,6-dibrompsnicilate) ·

In the first reaction step, the above-mentioned hydroxymethyl or 1-hydroxyethyl derivatives are converted to the corresponding trifluoromethanesulfonic acid esters (VII and VIII, where T * stands for trifluoromethanesulfonyloxyocapsule). This reaction is conveniently carried out at room temperature using trilluoromethanesulfonic anhydride as the reagent, in an inert solvent such as methylene chloride, in the presence of at least one equivalent of a tertiary amine such as pyridine or diieopropylethylamine.

In the second reaction step, the sulfonate moiety is replaced with an azide moiety, to form azidomethyl derivatives of formulas VII and VIII in which Y is azidoecupine. An excellent reagent for this purpose is tetramethylguanidine lumazide used in a slight excess. The reaction is carried out at a temperature of 0 to 25 ° C, preferably at a temperature of about 10 ° C, in an inert solvent such as chlorophone or methylene chloride.

In the third reaction step, the azidocycle is reduced to an amino group to give compounds of formulas VII and VIII in which Y is an amino group. A suitable reagent for this purpose is hydrogen sulfide in the presence of a tertiary amine such as triethylamine in an inert solvent such as chloroform. The reaction is carried out by introducing hydrogen sulphide gas into the reaction mixture at 0 to 50 ° C until the reduction is virtually complete, which takes about 3 to 4 hours at 25 ° C.

Finally, the amino and benzyloxy carbonyl groups are protected under standard conditions. As the reagent, for example, benzyl chloroformate is used in the presence of a tertiary amine, such as pyridine or opy, lam-diisopropylethylamine, and is operated in an inert solvent, such as methylene chloride, at 0 to 50 ° C, preferably at a lower temperature (0 to 50 ° C). The resulting compounds of formulas VII and VIII in which Y 'is benzyloxycarbonylamino are then subjected to further reactions according to the methods discussed in more detail above.

As mentioned above, some of the compounds of formulas I and II, generally those in which R @ 1 is hydrogen, have in vitro antibacterial activity. This efficacy is demonstrated by the measurement of minimum inhibitory concentrations of V / Ug / ml against a number of microorganisms. This measurement is performed according to the procedure recommended by the International Collaborative Study on Antibiotic Sensitivity Testing (Ericcon and Sherris, Acta). Pathologioa et Microbiologia Scandinav. Supp. 217 * Section B: 64-68 [1971]), using brain and heart infusion agar and re-inoculation devices. The contents of the tubes in which the respective cultures were grown overnight are 100-fold diluted and used as a standard inoculum. 20 ml of brain and heart infusion agar was added to each dish and 20,000 to 10,000 cells in 0.002 ml oca were coated on the surface of the agar. The test compound is used in twelve-fold dilutions, with an initial concentration of 200 µg / ml. Individual colonies are not taken into account when evaluating the plates after an eight hour incubation at 37 ° C. By sensitivity (minimum inhibitory concentration) of the test organism is meant the lowest concentration of the test compound capable of causing complete inhibition of microorganism growth when evaluated with the naked eye.

Those compounds of formulas I and II which exhibit this in vitro antibacterial activity are therefore useful as industrial antimicrobial agents, for example in water treatment, waste water and sludge disposal, paint protection and wood preservation, as well as disinfectants for topical applications. . When these compounds are used for topical application, it is often expedient to mix the active compound 8 with a non-toxic carrier, such as a vegetable or mineral oil or a softening cream. Similarly, the active ingredient may be dissolved or dispersed in liquid diluents or solvents such as water, alkanols, glycols or mixtures thereof. In most cases, compositions with an active compound concentration of about 0.1 to 10% by weight of the total composition are used.

As already mentioned above, the compounds of Formulas I and II are particularly valuable because they are potent inhibitors of microbial beta-lactam. This way these

CS 268 156 B2 compounds increase the antibacterial activity of β-lactam antibiotics (penicillins and cephalosporins) against numerous microorganisms, particularly O-lactam producing microorganismsBU · The ability of the above-mentioned compounds of formula I or II to increase the activity of β-lactam antibiotics can be demonstrated by wherein the minimum inhibitory konoentraoe one hand, the antibiotic alone, and secondly the actual compound of formula I or II (wherein R ¹ is hydrogen) · thus unmeasured MIC values are then compared with the minimum inhibitory concentrations found in use komblnaoa given antibiotic ee compound of pattern I or II, wherein R ¹ represents a hydrogen atom · When the antibacterial potency of the combination is significantly greater than would be expected based on knowledge of the individual potencies of the components occurs in the present case к increase efficiency · MIC values of the aforementioned combinations are measured using the method described by Barry and Sabath in Manual of Clinical Microbiology, ed. Lenette, Spauldlng and Truant, 2nd edition, 1974, American Society for Microbiology.

The compounds of Formulas I and II increase the antibacterial activity of β-lactam antibiotics in vivo, i.e., they reduce the amount of antibiotic required to protect the mouse from the otherwise lethal inoculum of a particular β-lactamase producing bacterium. To determine this potency, the mouse is induced by an acute experimental infection by intraperitoneal inoculation with a standardized culture of the test organism suspended in 5% mucin from the stomach of the pig · Infection intensity is standardized to give the mouse a lethal dose of the organism (lethal dose is minimal to reliably kill 100% of infected, not treated mice) · Groups of infected mice are then administered orally or intraperitoneally at different doses with the test compound in combination with an antibiotic · At the end of the test, the efficacy of the individual mixtures is evaluated by counting surviving animals from the group of animals . This efficacy is expressed as a percentage of surviving animals at a given dose or expressed as PD-θ, a dose protecting 50% of the test animals from infection. Compounds of formulas III and IV are tested for their in vivo activity in a similar manner except that they are administered alone and not in combination with other β-lactam antibiotics.

In order to determine whether a particular bacterial strain is susceptible to a particular compound of formula III or IV, it is not necessary to perform an in vivo test, but instead the minimum inhibitory concentration of a mixture of equal parts of a compound of formula I or II represents a hydrogen atom, and ampiollin or amoxicillin, as appropriate for each case.

Because of the ability of the compounds of Formula I and II to enhance the efficacy of β-lactam antibiotics against β-lactamase-producing bacteria, these compounds may advantageously be administered together with β-lactam antibiotics in the treatment of bacterial infections in mammals, particularly humans. In the treatment of a bacterial infection, a compound of formula (I) or (II) may be admixed with the .beta.-lactam antibiotic and both agents administered simultaneously. Alternatively, a compound of Formula I or II may be administered separately during treatment with a β-lactam antibiotic. In some cases, it is preferable to pre-medicate a patient with a compound of Formula I or II prior to initiating treatment with a beta-lactam antibiotic.

When using a compound of formula I or II to enhance the efficacy of a β-lactam antibiotic, the mixture of a compound of formula I or II in a β-lactam antibiotic is preferably administered in the form of a pharmaceutical composition which is included with standard pharmaceutical carriers or diluents. A pharmaceutical composition comprising a pharmaceutically acceptable carrier, a β-lactam antibiotic, and a compound of Formula I or II normally comprises about 5 to 80% by weight of a pharmaceutically acceptable carrier.

When the compounds of formula (I) or (II) are used in combination with a β-lactam antibiotic, these may be administered orally or parenterally, i.e., intramuscularly,

CS 268 156 B2 subcutaneously or intraperitoneally. Although the final decision on dosing in human medicine is a matter for the attending physician, the ratio of the daily doses of the compounds of Formula I or II to the β-lactam antibiotic will normally be in the range of about 1: 3 to 3: 1 (by weight). When using compounds of general formula I or II in combination with a ββ-lactam antibiotic, the daily dose of each component will normally range from about 10 to 200 mg per kilogram body weight by oral administration and about 10 to 40 mg per kilogram body weight by parenteral administration. In some cases, the attending physician may determine dosages outside these limits.

It is understood that some of the .alpha.-lactam compounds are effective when administered orally and parentally, while others are only effective when administered parenterally. If the compound of formula I or II is to be used concurrently (in admixture) with a .beta.-lactam antibiotic effective only If the compound of formula (I) or (II) is to be used concomitantly (in a mixture) with a .beta.-lactam antibiotic active in both oral and parental administration, compositions suitable for either oral or parenteral administration may be prepared. administration. In addition, the β-lactam antibiotic may be administered orally and parenterally, and the β-lactam antibiotic may be administered parenterally and orally.

Compounds of formula (III) or (IV) have the ability to hydrolyze and provide both a compound of formula (I) or (II) wherein R ^ * is hydrogen, as well as ampicillin or amoxicillin, thereby enhancing potency and broadening the spectrum of antibacterial activity thereof. ampicillin or amoxicillin, especially when administered orally.

When the antibacterially active compounds of formula III or IV according to the invention are used to combat bacterial infections in mammals, in particular humans, the compound is administered either alone or in admixture with pharmaceutically acceptable carriers or diluents. Said diluent or carrier is selected on the basis of the intended mode of administration. For oral administration, tablets, capsules, lozenges, troches, powders, syrups, elixirs, aqueous solutions and suspensions, etc. are used, in accordance with standard pharmaceutical practice. The ratio of the active ingredient to the carrier naturally depends on the chemical nature, solubility and stability of the active ingredient, as well as the dosage considered. In the case of tablets for oral administration, for example, lactose, sodium citrate and phosphoric acid salts are used as usual carriers. Various disintegrating agents, such as starch, and glidants, such as magnesium stearate, are usually used in the manufacture of tablets. Lactose and high molecular weight polyethylene glycols, for example polyethylene glycols having a molecular weight of from 2000 to 4000, are suitable diluents for the preparation of oral capsules. For the preparation of aqueous suspensions for oral administration, the active ingredient is combined with 8 emulsifiers or suspending agents. If desired, certain sweeteners and / or flavoring agents may also be added. For parenteral administration, including intra-muscular, intraperitoneal, subcutaneous, and intravenous administration, sterile solutions of the active ingredient are conveniently prepared, the pH of which is suitably adjusted and buffered. In the case of preparations for intravenous administration, the total concentration of all soluble fractions should be controlled so that the resulting preparation is isotonic ·

When using compounds of formula III or IV to combat bacterial infections, the daily doses will be within similar limits to other clinically used lactam antibiotics. Although the ultimate decision on the dosage used in human medicine will be made by the attending physician, the compounds of Formula III or IV will normally be used orally at doses ranging from about 20 to 100 mg per kilogram of body weight per day and parenterally at doses ranging from about 10 to 100 mg. 100 mg per kilogram of body weight per day, usually in divided doses. In some cases, the physician may determine dosages outside the above range as appropriate.

CS 26θ 156 B2

The invention is illustrated by the following non-limiting Examples, unless otherwise indicated, proton nuclear magnetic resonance spectra (PNMR) were measured at 60 MHz.

Example 1

Benzyl-6H-bromo-6A- (benzyloxycarbonylaminomethyl) -penioilanoate and -6β-bromo-β- (benzyloxycarbonylaminomethyl) penicilane ·

To a solution of 108.73 g (0.242 mol) of benzyl 6,6-dibromopeniailate in 600 ml of dry tetrahydrofuran cooled to -78 ° C was added 83.5 ml of a 2.9M ethereal methylmagnesium bromide solution. After fifteen Iminutovém stirring at -78 ° C was added over 10 minutes 27 E (0.121 mol) benzyloxykarboxamidomethyl aoetátu-dissolved in 200 ml of dry tetra _T dihydrofuran. After stirring at -78 ° C for 1 hour, the reaction was quenched by the addition of 14.52 ml of acetic acid, warmed to room temperature and the volatiles evaporated in vacuo below 35 ° C. The residue was dissolved in ethyl acetate, washed with 100 ml. ml of water, 100 ml of aqueous sodium bicarbonate solution and twice with 100 ml of water each, dried over sodium sulfate and concentrated in vacuo. 113 g of an oily product are obtained which is chromatographed on a column of 1.2 kg of silica gel. The column is eluted first with 6 liters of ore in equal parts of hexane and chloroform and then with chloroform. The first 6 liters of eluate are discarded and the next eluate is collected in 25 ml fractions. Fractions 181-190 are concentrated. The PNMR of the residue indicates the presence of benzyl-6C-bromo-6β- (benzyloxycarbonylaminomethyl) penicillanate · PNMR (deuterochlorophon, tetramethylsilane, i-values):

* 1.37 (3H, s), 1.57 (3H, s), 3.86 (2H, d, J = 6 Hz), 4.42 (1H, s), 5.06 (2H, s) ,

5.12 (2H, s), 5.52 (1H, s), 7.25 (10H, s),

Fractions # 201-249 ee thickened · PNMR spectrum of this residue indicating the presence of benzyl 60-bromo-6c - (benzyloxycarbonylaminomethyl) penicillanate · PNMR (deuterochloroform, tetramethylsilane, values (()):

1.36 (3H, s), 1.60 (3H, s), 3.90 (2H, d, J = 6.2 Hz), 4.47 (1H, s), 5.07 (2H, s), 5.14 (2H, s), 5 · 4Ο (1H, t, J = 6.2),

5.47 (1H, s), 7.28 (5H, s), 7.30 (5H, s).

The product from fractions Nos. 171-240 is combined and concentrated to give 22 g of a foamed material as used in Example 2 below.

Example 2

Benzyl-6β- (b enzyloxycarbony laminomethyl) penicilate To a solution of 22 g (0.0413 mol) of the title products) a mixture of epimers) in 100 ml benzene ee add 32.7 ml (0.124 mol) The reaction mixture was heated under reflux for 2 hours under nitrogen, then concentrated in vacuo and the oily residue triturated four times with 100 ml of hexane each time. The residual viscous oil was taken up in 70 ml of ether and crystallized within 1 hour. 8.1 g in two portions of the title compound · PNMR (deuterochloroform, tetramethylsilane, cT values):

1.37 (3H, s), 1.57 (3H, s), 3.58 (3H, m),

4.34 (1H, s), 5.04 (2H, s), 5.12 (2H, s),

5.33 (1H, d, J = 4Hz), 7.32 (1H, s).

Example 3 1'-Benzyl-60- (benzyloxycarbonylaminomethyl) -penicillanate and 1 (b-Benzyl-6β- (benzyloxycarbonylaminomethyl) -penicillanate).

CS 268 156 B2

To a solution of 4 * 54 s (0.01 mol) of the title compound in 70 ml of ethyl acetate was added 2.02 g (0.01 mol) of m-chloroperbenzoic acid in 30 ml of ethyl acetate. It is then washed once with 50 ml of saturated sodium bicarbonate solution and twice with 50 ml of water each, dried over sodium sulphate and concentrated in vacuo to a viscous oily residue. This oil is dissolved in 50 ml of ether and 10 ml of chloroform. Scratching the vessel walls with a glass rod gives crystallization of the title C oxidu oxidu oxide. The yield of the product melting at 123-124 ° C is 2.2 g.

PKMR (deuterochloroform, tetramethylellan, values ^) ’

1.22 (3H, e), 1.51 (3H, a), 3.7 (ЗН, в),

4.34 (1H, d), 4.63 (1H, d, J = 4Hz),

5.13 (2H, a), 5.22 (2H, a), 5.50 (1H, в),

7.34 (5H, e); 7.40 (5H, a).

Concentration of the mother liquor under vacuum to dryness afforded the title D -oxide as an oil in a yield of 2.5 g.

PNMR (deuterochloroform, tetramethylsilane, values of i):

1.05 (3H, s), 1.60 (3H, s), 3.8 (3H, m), 4.63 (1H, s), 4.73 (1H, d, J = 4 Hz),

5.13 (2H, s), 5.23 (2H, q), 5.70 (1H, m),

7.35 (5H, b) ₉ 7.39 (5H, s).

EXAMPLE 4 1 (Benzyloxybenzyl) -6- (benzyloxycarbony laminomethyl) -penicilane

To 2.3 g (4.9 mmol) of the title β-oxide in 100 ml of chloroform was added 0.607 g (4.9 mmol) of 1,5-diazabicyclo [4.3.0] non-5-ene . The reaction mixture is stirred at room temperature for 15 minutes, then diluted with 50 ml of 1N hydrochloric acid and the layers are separated. The organic layer is washed twice with 50 ml of water each time, dried over sodium sulfate and concentrated in vacuo. 2.3 g of an oily residue are obtained, which is chromatographed on a silica gel column. Elution is carried out with a 4: 1 mixture of chloroform and ethyl acetate, collecting 20 ml fractions. Frakoe Nos. 41 to 70 are combined and concentrated in vacuo. 0.9 g of the title compound is obtained in the form of a viscous oil,

PNMR (deuterochloroform, tetramethyleilane, values of i):

1.03 (3H, s), 1.60 (3H, s), 3.67 (3H, m), 4.46 (1H, s), 4.88 (1H, m), 5.08 (2H) , s), 5.17 (2H, q), _t 5.39 (1H, m), 7.32 (5H, s)

7.37 (5 H, s),

Example 5

Benzyl-όβ - (benzyloxycarbonylaminomethyl) -penicillanate 1,1-dioxide

To a solution of 1.0 g (0.0176 mol) of the title compound of Example 2 in 200 ml of ethyl acetate cooled to 0-5 ° C was added 10.68 g (0.0528 mol) of m-chloroperbenzoic acid cyeeline. The reaction mixture was warmed to room temperature, stirred for 6 hours, recooled to 0-5 ° C and diluted with 50 mL saturated sodium bicarbonate solution. The organic layer was separated, washed twice with 50 ml each saturated sodium bicarbonate solution and twice with 50 ml water each, dried over sodium sulfate and concentrated in vacuo. 6.6 g of a viscous oil are obtained which is chromatographed on 250 g of silica gel with ethyl acetate / chloroform 1: 19 as the eluent, collecting 25 ml fractions. Frakoe Nos. 44 to 150 are combined and concentrated in vacuo. This gave 7.6 g of the title compound as a white resin foam.

PNMR (deuterochloroform, tetramethylsilane, К values):

CS 268 156 B2

1,25 (3H, ^s )> 1.49 (3H, c), 3.98 (3H, 4.45 (Ш, 4.59 (1 H, d,. J = 4 Hz). 5.09 (2H, в), 5.19 (2H, q); 5.36 (1H, years signal), 7.36 (11H, s) • ] Ex klad 6

Benzyl-6 Cd - (benzyloxycarbonylaminomethyl) penicllanate.

Using the procedure described in Example 4, 3.3 g (6.79 mmol) of 1,1-dloxlde of the above title were converted to the title compound of this example. The crude product yield is 3.1 g. Purification is carried out by column chromatography on 150 g of silica gel, eluting with ethyl acetate / chloroform (1: 9). Fractions of 20 ml are collected. Fractions 26-37 are combined and concentrated in vacuo to give the purified title product as a viscous oil which crystallizes on standing. The yield of pure product melting at 112-113 ° C was 1.9 g.

PNMR (deuterochloroform, tetramethylsilane, values):

1.20 (3H, s), 1.49 (ЗП, s), 3.65 (3H, m),

4.32 (1H, s), 4.59 (III, m), 5.07 (2H, s), 5.14 (2H, q), 5.30 (1H, broad signal),

7.32 (10 H, s).

The above product was also obtained by further oxidizing the title compound of Example 4 with an excess of m-chloroperbenzoic acid according to the procedure described in Example 5.

Example 7

6β - (Amlnomethyl) foamylionic acid 1,1-dloxide

1.9 g of the title product 5, 40 ml of tetrahydrofuran, 40 ml of water and 1.9 g of 10% palladium on carbon are mixed and the mixture is hydrogenated for 1 hour under a pressure of 50 psi. The catalyst was filtered off and tetrahydrofuran was evaporated from the filtrate in vacuo.

The aqueous layer was washed with 30 mL of ethyl acetate and freeze-dried. A white powdery material was obtained which, by trituration and 5 ml of water, gave 0.26 g (first crop) of crystalline product. A second crop of 0.14 g of product crystallized in the mother liquor after addition of 10 ml of acetone and a third crop of 0.35 g was obtained by evaporating the second mother liquor to a volume of 2 ml and adding 50 ml of acetone. 0.75 g.

PNMR (250 ΜΠζ, deuterlumoxide, 2,2-dimethylsllapentane-5-sulfonate, values (Γ):

1.47 (3H, 6). 1.59, (311, s), 3.74 (2Π, m), 4.36 (1H, td,

J »4, 5.5 Hz), 4.45 (1H, s), 5.17 (III, d, J 4 Πζ).

Example 8

1.1-Dloxld 6 cC- (Amino-ethyl) -pentillanic acid

Using the procedure described in the previous example, 1.7 g of the title compound of Example 6 was converted to the title product, except that the crystalline product was obtained directly after extraction with ethyl acetate by concentration in vacuo. Yield 0.7 g.

PNMR (250 MHz, deuterlumoxide, 2,2-dlmethylsllapentane-5-sulfonate, J-values):

1.44 (3H, s), 1.59 (3Π, s), 3.63 (2H, d, J with 5.5 Hz), 4.07 (1Я, td, J »2.5, 5.5 Hz), 4.31 (III, s), 5.06 (1Π, d, J = 2).

CS 268 156 B2

Example 9

Bensyl-6β-bromo-6C-Trifluoromethanesulfonyloxymethyl-peniolanate

To a solution of 3.15 mL of trifluoromethanesulfonic anhydride in 20 mL of methylene chloride at room temperature was added a solution of 6.232 g (15.6 mmol) of benzyl 6/3-bromo-6α- (hydroxymethyl) -polyl lineate and 1.89 µl of pyridine in 20 mL. The methylene chloride was evaporated under reduced pressure and the residue was partitioned between ethyl acetate and water. The ethyl acetate phase was separated and the aqueous phase was extracted with further ethyl acetate. The combined ethyl acetate solutions were washed first with sodium bicarbonate pH 8.3 and then brine. After drying over anhydrous sodium sulfate, the resulting solution was evaporated under reduced pressure to dryness to give 8.296 g of the title compound as an orange solid. PNMR (deuterochloroform, tetramethylsilane) (d):

1.41 (s, 3H), 1.63 (s, 3H), 4.51 (s, 1H),

4.87 (s, 2H), 5.14 (β, 2H), 5.44 (s, 1H),

7.30 (s, 5H)

Example 10

Benzyl-6-azidomethyl-6? -Brompenloilaneate

To a solution of 8.296 g (15.6 mmol) of benzyl β-Bromo-6C-trifluoromethylsulfonyloxymethylpenioilate in 50 mL of chloroform at 10 ° C was added 2.96 g (18.7 mmol) of tetramethylgusidinidine azide. The reaction mixture is stirred for 1 hour, then concentrated to 1/3 of the original volume and filtered through a pad of silica gel. The silica gel pad is washed with 100 ml of 10% ethyl acetate in chloroform and the eluate is evaporated. amber oil ·

PNMR (deuterochloroform, tetramethylsilane, values):

1.38 (s, 3H), 1.61 (s, 3H), 3.96 (s, 2H),

4.53 (b, 1H), 5.17 (in, 2H), 5.40 (e, 1H),

7.34 (t, 5H).

Example 11

Benzyl dj -bromo-6β-trifluoromethanesulfonyloxymethyl-penicillanate

Using the procedure described in Example 9, a solution of 0.548 g (1.4 mmol) of benzyl 6-tert-bromo-6β-hydroxymethylpenicilate in 4 ml of methylene chloride containing 0.17 ml of pyridine is treated with a solution of 0.42 ml of trifluoromethanesulfonic anhydride in 3 ml. ml of methylene chloride to give 641 mg of the title compound as an amber oil.

PNMR (deutero-chloroform, tetramethylsilane, δ *) x

1.43 (β, 3H), 1.62 (s, 3H), 4.52 (в, 1Ή),

4.88 (q, 2H), 5.19 (in, 2H), 5.62 (β, 1H),

7.35 (в, 5H) ·

Example 12

Benzyl-6C-bromo-6β-azidomethylpenioilate

To a solution of 641 mg (1.2 mmol) of benzyl-bromo-6β-trifluoromethanesulfonyloxymethylpenioilanoate in 10 mL of chloroform at 10 ° C was added 229 mg (1 * 2 mmol) of tetramethylguanidiniumazide, stirred for 1 hour and then evaporated under reduced pressure ·

The oily residue is filtered through a pad of silica gel, which is then eluted with 10% ethyl acetate in chloroform. Evaporation of the eluate gives 420 mg of the title compound as an amber oil.

CS 268 156 B2

PHMR (deut.roohlorofoXBitetraB.thyleilan, dd values) i

1.0 (e, JH), 1.61 (·, 3H), 3.91 (·, 2H),

4.48 (β, 1H), 5.15 (·, 2H), 5.57 (β, 1H),

7.37 (t, 5H).

Example 13

Benzyl-6C- (endnomethyl) 76β-bromopenol aclanate

To a vigorously stirred solution of 541 mg (1.3 mmol) of benzyl 6cG-azidomethyl-6H-bromobenicenilate and 0.71 ml (4 equivalents) of triethylamine in 10 ml of chlorofoxra were added hydrogen sulfide gas for 1 hour. in vacuo to a red-colored oily residue which, according to NMR spectroscopy, is the desired product contaminated with triethyl amine »

PNMR (deut его chloroform, to tramyl thylsilane, values <T) p

1.39 (,, 3H), 1.64 (,, 3H), 3.35 (,, 2H), 4.51 (,, 1H), 5.16 (,, 2H), 5.35 (а) , 1H), 7.33 (α, 5H).

Example u

Benzyl-6'-bromo-6'-benzyloxycarbonylaminomethylpenioilate

To a solution of benzyl chlorphormiate т 5 ml methylene chloride and add a solution of 239 mg (0.6 mmol) of benzyl 6-C-aminomethyl-1-beta-bromobenzene cllanate and 0.14 ml of pyridine in 5 ml of methylene chloride over 5 minutes using a syringe. The mixture is stirred under ice-cooling for 75 minutes under nitrogen, then evaporated under vacuum and the residue is taken up in ethyl acetate / water. The pH is adjusted to 2.9 with dilute hydrochloric acid, the ethyl acetate phase is separated, extracted with dilute sodium bicarbonate solution. pH 8.1), washed with brine and dried over anhydrous sodium sulfate. After evaporation under reduced pressure, the residue (312 mg) is taken up in chloroform and chromatographed on a 15 g silica gel column (column dimensions 1.4 x 20 cm), elution with 5% ethyl acetate in chloroform · 4 ml fractions are collected · Fractions 14-14 to 27 are combined and evaporated under reduced pressure to give 168 g mg of the title product · PNMR (deutero-chloroform, tetramethylsilane, values <5 *) x The spectrum corresponds to the title compound and is identical to that of the same compound prepared in Example 1.

Example 15

Benzyl-6β - (benzyloxycarbonylaminomethyl) penicillanate

A solution of 0.25 ml of tri-n-butyltin hydride and 168 mg (0.31 mmol) of benzyl 6-bromo-6-D- (benzyloxycarbonylaminomethyl) penloilanoate in 4 ml of benzenes was heated at reflux for 2 hours, then benzene was added. evaporate in vacuo and the residue triturated three times with 2 ml of hexane each. The residue is then taken up in ethyl acetate in water, the ethyl acetate phase is separated, washed with brine and dried over anhydrous sodium sulphate. After evaporation in vacuo, 101 mg of oily material is obtained which is shown in column 4 g of silica gel (column dimensions). 1 x 11 cm), which is prepared in chloroform and washed with 5% ethyl acetate in chloroform. 4 ml fractions are collected. Frakoe 3 to 5 ee were combined and, after evaporation of the ee, 66 mg of the title product was obtained which, according to PNMR, was identical to the product prepared in Example 2.

Example 16

Bsnzyl-6? - (benzyloxycarbonylaminomethyl) penioilanate and benzyl-6? - (benzyloxycarbonylaminomethyl) penioilanate

CS 268 156 B2

The required Grigard reagent is prepared essentially according to the method described by DiNlnno et al., J. Org. Chem · 42 · pp. 2960 - 2965 (1977) · Dissolve benzyl 6-iodo-iodo-lalanate in 75 ml dry tetrahydrofuran »cool the solution from -78 ° 0 under dry nitrogen and add dropwise thereto 5» 6 ml of JM methylmagnesium bromide in ether. After stirring for another fifteen minutes, a solution of 1.87 g of benzyloxycarbonylaminomethylacetate in 25 µl of dry tetrahydrofuran was added in one portion, stirred again at -78 ° C for 15 minutes, then 2 ml of acetic acid was added thereto, the reaction mixture was warmed. to 0 ° C and evaporated in vacuo · The residue is shaken between 250 ml of ethyl acetate and 50 ml of water · The organic layer is separated, washed once with 100 ml of saturated sodium bicarbonate solution and twice with 100 ml of sodium chloride solution each time, dried over sodium sulfate. £. Nym and evaporated in vacuo to an oily residue weighing 7.3 g · The oil is chromatographed on 250 g of silica gel eluting with ethyl acetate-chloroform (1: 10) _t collecting fractions of 20 ml · Prakce no. 20 to 24 contain 1.3 g of oily by-product and fractions 25 to 34 contain 0.62 g of the mixture of the - and db-isomers (3 t 2) in the title (according to PNMR) · Prakoe No. 35 to 60 contain 2 , 2 g of a mixture of (3 t-1) isomers in the title.

Example 17

Benzyl-6'-bromo-6 (b - (1R-trifluoromethanesulfonyloxyethyl) penicllanate

20.28 g (0.0489 mol) of benzyl 4'-bromo-6β- (1R-hydroxyethyl) penicillanate (DlHlnno et al., J. Org. Chem. 42, p. 2960-2965, 1977) was dissolved in 400 g. ml of dichloromethane, the solution is cooled in a mixture of ice and acetone and then, while maintaining the temperature at &lt; 5 ° C, 7.91 ml (2 equivalents) of pyridine is added dropwise and then 11.58 ml (1.4 equivalents) of trifluoromethanesulfonic anhydride ·

The resulting mixture was stirred at 0 ° C for 30 min, then diluted with dichloromethane, washed successively with saturated sodium bicarbonate solution, water and saturated sodium chloride solution, evaporated in vacuo and the residue diluted 1: 1 with hexane. 18.71 g of the title compound. .

PNMR (deuterochloroform, tetramethylellan, values 0):

1.42 (3H, e), 1.55 (3H, d, J = 6Hz),

1.66 (3H, in), 4.58 (1H, in), 5.20 (2H, e),

5.34 (1H, q, J = 6Hz), 5.56 (1H, s),

7.38 (5H, a). .

Example 18

Benzyl-6 &apos; -bromo-6 &apos;-( 1S-azidoethyl) penicillanate

34.4 g (0.0628 mol) of the title compound are dissolved in 400 ml of dichloromethane, 25.0 g (1.4 equivalents) of tetrabutylammoniumazide in 100 ml of dichloromethane are added dropwise, and the mixture is stirred at room temperature for 1 hour. Deň: 22 ° C. The dichloromethane is evaporated in vacuo and the oily residue is filtered through 500 g of silica gel in a sintered glass funnel. The silica gel was eluted with 2 liters of dichloromethane and the eluate was evaporated to give 27 g of the title product as a light yellow solid.

Yellow Oil ·

PNMR (deuterochloroform, tetramethylslan, values) x

1.40 (3H, s); 1.53 (3H, s);

1.57 (3H, s); 4.10 (1H,

4.47 (1H, s), 5.19 (2H,

7.34 (5H, s) · d, J = 6.3 Hz), q, J = 6.3 Hz), s), 5.61 (1H, s).

Example 19

Benzyl-6β - (1S-aminoethyl) -6C-bromopeniollanate

27.0 g (0.0613 mol) of the title compound are dissolved in 300 ml of chloroform, 35 ml (4 equivalents) of triCS 268 156 B2 ethylamine are added and the gas is stirred for 3> 5 hours. The reaction mixture was purged with nitrogen for 0.5 hours, after which the chloroform was evaporated in vacuo. The oily residue was partitioned between ether and 1H hydrochloric acid (200 ml) and the ether layer was extracted three times with 200 ml of fresh 1N hydrochloric acid each. dilute the layers with ethyl acetate and adjust the pH of the two-phase system to 8.5. Partition and evaporate to give 6.34 8 of the title compound as an oil. The original ether layer is diluted with water and the pH is adjusted to 8.5. The ether layer is separated and evaporated to give a further residue. 6.0 g of the title compound ·

PNMR (deutero-chloroform, tetramethylsilane, У values) t

1,28 (зн, d, J 6j , 3 ha), 1.40 (3H, я), 1.58 (зн, e), 3.34 (1H, q, J = 6.3Hz), 4 »45 (1H, »), 5.16 (2H,?), 5.54 (1H,?). 7.33 (5H, E). Ex : к i and d 20

Benzyl-6β - (LS-benzyloxycarbonylamino-ethyl) -6-C-bromobenzicilate

Dissolve 5.10 mL (1.2 equivalents) of benzyl chloroformate in 20 mL of dichloromethane, cool the solution in ice-acetone, and add a solution of 12.34 g (0.02980 mol) of the title compound dropwise. of the preceding example in 50 ml of dichloromethane containing 7.78 ml (1.5 equivalents) of diisopropylethylamine. During the addition, the temperature of the reaction mixture was kept at 0-5 ° C by cooling in ice-water. The organic layer was separated, washed with water and saturated brine, and evaporated. 17 g of the title product are obtained in the form of a pale yellow oil.

PNMR (deuterochloroform, tetreunethylflilan, 0 values) ) « 1.35 (3H, e), 1.39 (3H, d)> 1.55 (3H, with) 4.26 (1 H, m), 4.45 (1H, e) _t 4.97 (1H, (d) 5.08 (2H, d), 5.14 (2H, E)" 5.53 (1H, E) 7.33 (M, N).

Example 21

Benzyl-4- (1S-benzyloxycarbonylaminoethyl) foaming ilate

16.33 g (0.0298 mol) of the title product are dissolved in 200 ml of benzene, 23.5 l (3 equivalents) of trin-butyl tin hydride are added and the mixture is refluxed for 3 hours. The benzene was evaporated in vacuo and hexane was added to the residue. The title compound crystallizes in 9> 26 g.

PNMR (deuterochloroform, tetramethylsilane, values of i):

1.39 (3H, d, J-6 Hz), 1.40 (3 H, e), 1.63 (3H, в), 3.62 (1H, dd, J »4, 11 Hz), 4.12 (1H, m), 4.39 (1H, я), 4.81 (1H, d, J - 8 Hz), 5.08 (2H, a), 5.17 (2H, e), 5.32 (1H, d, J-4 Hz), 7.32 (5H, s), 7.36 (5H, E). P г j í к 1 and d 22

Benzyl-6 (3- (1S-benzyloxycarbonylaminoethyl) penicillanate 1,1-dloxide

Using the procedure described in Example 50, the title compound of the previous example (4.0 g, 0.0085 mol) was converted to the title product of this example.

CS 268 156 B2

This product ее licks те thyme 4.25 g те white foam form.

PKMR (deutero-chlorofox, tetremethyleilane, δ values) «

1.21 (3H, a), 1.33 (3H, d, J = 6Hz),

1.48 (3H, a), 4.20 (1Я, в), 4.35 (1H, в),

4.40 (1Я, a), 4.53 (1H, d, J = 4.5 Hz), 5.08 (2H, a), 5.17 (2Я, q), 5.45 (1Я, d) J - 7 Hz), 7.30 (5H, a), 7.35 (5H, a).

Example 23

6,5- (13-Aminoethyl) penicillanic acid 1,1-dioxide

Using the procedure of Example 7, the title compound (175 mg) was converted to the title product using 180 mg of 5% palladium on carbon catalyst. After separation of the catalyst, the filtrate, along with aqueous tetrahydrofuran washings, was freed of liquid. tetrahydrofuran by evaporation in vacuo and the product isolated by lyophilization of the aqueous residue over noo.

IR (KBr) 1765 om t ^_1.

PNMR (deuterium oxide, 2,2-dimethylsilapentane-5-eulfonate, <f) t 1.47 (3H, a), 1.53 (3H, d),

1.60 (3H, a), 4.35 (2H, α), 4.37 (1H, a),

5.12 (1H, d, J = 4Hz),

Example 24 _l-f l-benzyl 6dL- dioxide (S-benzyloxycarbonylamino) penicilenátu

4.25 g (0.0085 mol) of the title compound of Example 22 are dissolved in 100 ml of dichloromethane, 0.96 ml (1 equivalent) of 1,5-diazabicyclo [4.3.0] non-5-ene is added dropwise. The mixture was stirred at 25 ° C for 3 minutes, then quenched by the addition of 1 mL (2 equivalents) of glacial acetic acid. The resulting mixture was diluted with dichloromethane, washed successively with dilute hydrochloric acid (pH 2.5), water and saturated sodium chloride solution, and evaporated in vacuo. The title compound as a white pin · Column chromatography on silica gel using dichloromethane / ethyl acetate (9: 1) as eluent gave 2.83 g of the purified title product as a white pin · PNMR (deuterochlorofoxm, tetramethylsilane) , values & ):

1.21 (3H, в), 1 · 35 (зн, d, J = 7 Hz), 1 51 (3H, ·). 3.78 (1Н, dd, J - 2.4 Hz) 4.27 (1 H, m), 4.36 (1Н, a), 4.56 (1H, d, J - 2 Hz) 5.09 (2H, a), 5.16 (2H, q) 7.3 (ЮН, в). P i i 1 and d 25

6 of - (1S-Aminoethyl) penicillanic acid 1,1-dioxide

4.5 g of 5% palladium on carbon are suspended in 50 ml of water and

It is prehydrogenated at 25 ° C / 0.35 MPa for 1.5 hours. 2.83 g of the title compound are dissolved in 50 ml of ethyl acetate, the solution is added to a prehydrogenated aqueous catalyst slurry and hydrogenated at 25 ° C / 0.3 MPa for 0.5 h. The catalyst is filtered off and the aqueous layer is separated . When the aqueous layer of the crystal is concentrated, the title product is present in three portions (1.1 g total weight).

PNMR (deuterium oxide, 2,2-dimethylsllapentane-5-sulfonate, t-values):

1.47 (3H, s), 1.52 (3H, d),

1.62 (3H, s), 4.05 (2H, m), 4.28 (1H,),

CS 268 156 B2

5.10 (1H, d) · δ (KBr) x 1787 ов

Example 26

Benzyl-6-bromo-6C- (1-trifluoromethylsulfonyloxyethyl) penicillanate

Using the procedure of Example 17, 8.90 g (0.0214 mol) of benzyl 4-β-bromo-6e (1-hydroxymethyl) penioilanoate (a mixture of ephneres on the side chain) of DiNinno and the above oitaoi were converted to the product. · A first portion of a 4.70 g solid product, which is predominantly IR-epimer on the side chain, crystallizes after a mixture of equal portions of hexane and ether (if desired, this product can be worked up according to the procedures of Examples 25-30). to form the title compound of Example 32) A second portion of the solid product of 2.40 and a third portion of 1.60 g (oily) are composed of a mixture of epimers of the title compound, containing predominantly the 1S-eplmer on the side chain,

PNMR (dmiterochloroform, tetramethylsilane, hodnoty values) x

1.41 (3H, s), 1.65 (3H, s), 1.78 (3H, d, J * 6.5 Hz), 4.56 (1H, s), 5.22 (2H, s) ), 5.37 (1H, q, J = 6.5 Hz), 5.48 (1H, s),

7.4 (5H, s).

Example 27

Benzyl- [1- (1-azidoethyl) -6 (1H) -brompenicilate

4.0 g (0.0073 mol) of the second and third portions of the title product (predominantly 1S-eplmer in the side chain) were converted to the title compound (predominantly 1R-eplmer in the side chain) by the procedure of Example 18. . This product was isolated as a pale yellow oil in a yield of 2.91 g. PNMR (deuterochloroform, tetramethylsilane, values (!):

1.40 (3H, s), 1.56 (3H, d, J = 6.5 Hz), 4.00 (1H, q, J = 6.5 Hz), 4.48 (1H, s), '

5.16 (2H, s), 5.34 (1H, s), 7.32 (5H, s).

Example 28

Benzyl-6c- (1R-aminoethyl) -60-bromo-penicillanate and benzyl-6R- (1S-aminoethyl) -6 (1-bromo-penicillanate). Following the procedure of Example 19, the title compound of the previous example (2.91 g, 0.0066 mol of predominantly the 1R-epimer on the side chain) was converted to the title products of this Example. After removal of the chloroform used as the reaction solvent, the residue was partitioned between ether and 1N hydrochloric acid (200 ml). The ether layer was separated and extracted once with 200 ml of 1N hydrochloric acid. The combined aqueous layers were overlaid with ethyl acetate and the pH adjusted to 8.5. The aqueous layer was separated and extracted with fresh ethyl acetate. The ethyl acetate layers were combined and evaporated to give 1.53 g of an oily mixture of the title products (predominant 1R-isomer). The isomers were separated by column chromatography on 200 g of silica gel using a mixture of equal parts of dichloromethane and ethyl acetate as eluant, The column was monitored by thin layer chromatography in the system dichlonoethan - ethyl acetate 3: 2 clean faster moving fraction (R *. ³ 0.52) were combined and evaporated to give 0.172 g of the title S-isomer, which was optionally converted by the procedures described in Examples 20 to 23 to the title compound of Example 25. 0.45 g was obtained from the medium chromatographic fractions. g a mixture of the title compounds, which may optionally be re-chromatographed to provide additional pure products. From the combined pure, slower-running fractions (Rf = 0.47), 0.674 g of the title side-chain 1R-epimer was obtained.

CS 268 156 B2

PKMR (deuterochloroform, ethyl silon tetras, values ^):

1.29 (3H, d, J 6 Hz), 1.38 (3H, s),

1.64 (3H, s), 3.29 (1H, q = 6 Hz),

4.51 (1H, s), 5.17 (2H, s), 5.38 (1H, s), 7.33 (5H, a).

Example 29

Benzyl-бЛ-отгот-ба-CR-benzyloxycarbonyleminoethyl) - foams or ropes

Using the procedure described in Example 20, 0.674 g (0.0016 mol) of the title R-isomer of the previous example was converted to 0.877 g of the title compound having an Rf of 0.85 (a mixture of equal parts of dichloromethane and ethyl acetate). .

Example 30

Benzyl-6- (1R-benzyloxycarbonylaminoethyl) foam cllanate

0.877 g (0.0016 mol) of the title compound was converted to the title product by the procedure of Example 21. After evaporation of the benzene used as the reaction solvent, the residue was triturated four times with 50 ml of hexane each to give 627 mg of crude product as an oil. This oil was chromatographed on silica gel using chloroform / ethyl acetate (19: 1) as eluent. 569 mg of purified title product is obtained.

PNMR (deuterochloroform, tetramethylsilane, $ values):

1.10 (3H, d, J ж 6, 5 Hz: ), 1.36 (3H, a] 1. 1.60 (3H, a), 3.41 (1H, dd, Jn 4, 11 Hz) 4.16 (1H, m), 4.43 C1H, s), 5.08 (2H, with)" 5.13 (2H, a), 5.32 (1H, d, J (4 Hz), 7.29 (5H, a), 7.33 (5H, with).

Example 31

Benzyl-6β- (1R-benzyloxycarbonylaminoethyl) penicillanate 1,1-dioxide

0.569 g (0.0012 mol) of the title compound was converted to 0.681 g of the title product by the procedure of Example 5.

PKMR (deuterochloroform, tetramethylsilane, ^ values):

1,22 (3H, with), 1.27 (3H, (J), 1.47 (3H, a), 3.94 (1H, dd, J «4, 12 Hz), 4.42 (1H, a) 4.48 (2H, m), 5.04 (2H, a), 5.13 (2H, q); 5.40 (1H, d, J ¹ «8) 7 25 (5H, a) . 7.32 (5H, with). Ex í к 1 and d 32

6β - (1R-Emino-thylenepentilicanoic acid) 1,1-dioxide

202 mg of the title compound of the previous example were used

400 mg of 5% palladium on carbon are converted to 50 mg of the title product. IC (KBr-technology) :: 1788 сш ¹ .

PNMR (deuterlumoxide, 2,2-dimethylsilapentene-5-sulfonate, ^ values):

1.45 (3H, a), 1.51 (3H, d),

1.57 (3H, [alpha]), 4.27 (1H, [alpha]), 4.33 (1H, [alpha]),

4.85 (1H, d), 515 (1H, d, J = 4 Hz).

Example 1 a d 33

1,1-Dioxide benzyl-6 &apos;-( 1H-benzyloxycarbonylaminoethyl) pentanilate

CS 268 156 B2

368 sg of the title compound of Example 31 were converted to 285 ng of the title product using Example 19 using chloroform / ethyl acetate (19: 1) as the eluent. PNMR (deuterochlorophons, tetramethylsilane, X-values):

1.23 (3H, s), 1.33 (3H, d, J * 6.5 Hz),

1.50 (3H, s), 3.61 (1H, dd, J * 2.9 Hz), 4.28 (IH, m), 4.34 (IH, a), 4.67 (IH, d) 1 J (2 Hz), 4.98 (1H, d), 5.07 (2H, s), 5.18 (2H, q), 7.30 (5H, s), 7.35 (5H, s).

Example 34

6? (1R-Aminoethyl) -caproic acid 1,1-dioxide

285 mg of the title compound was converted to 132 mg of the title product by the procedure of Example 32. IR (KBr technilce): 1768 cm ^1st

PNMR (deuterlunoxide, 2β-diethylsllapentene-S-sulfonate, сУ values):

1.47 (3H, s); 1.54 (3H, d);

1.61 (3H, a), 4.03 (2H, m), 4.44 (1H, s),

5.10 (1H, d, J = 2).

Example 35

60- (benzyloxycarbonylaminomethyl) penicillanic acid

Method A

II, 2 g of the title compound of Example 6 was partially hydrogenated in 70 ml of tetrahydrofuran and 50 ml of water for 30 minutes in the presence of 6 g of 10% palladium on carbon at 50 psi. The catalyst was filtered through a pad of diatomaceous earth, tetrahydrofuran was distilled off in vacuo, and the aqueous residue was extracted with 100 ml of ethyl acetate. The organic layer was separated, dried over sodium sulfate and evaporated. 3.0 g of the title product are obtained in the form of a foam.

PNMR (deuterochloroform, tetramethylsilane, values of 5):

1.40 (3H, s), 1.55 (3H, s), 3.70 (3H, m),

4.31 (1H, s), 4.58 (1H, m), 5.04 (2H, s),

7.24 (5 H, s).

Concentration of the aqueous layer yields 3.1 g of crystalline 1,1-dioxide - (aminomethyl) foamic acid, whose PNMR spectrum is identical to that of the fully hydrogenated product of Example 8.

Method В

3.0 g (11.45 mmol) of the product of the title compound of Example 8 are dissolved in 100 ml of a mixture of equal parts of water and methanol. The pH was adjusted to 8.3-8.7, and 1.79 g (12.59 mmol) of benzyl chloroformate was added dropwise over several minutes. After stirring briefly, the pH is adjusted to 6.0 with 1N hydrochloric acid and the tetrahydrofurene is distilled off in vacuo. The aqueous residue was extracted with 30 mL of ethyl acetate and the extract was discarded. After addition of 50 ml of fresh ethyl acetate, the pH is adjusted to 1.8 with 1N hydrochloric acid and the aqueous layer is extracted with 50 ml of fresh ethyl acetate. The organic layer was combined with the extract, washed once with 50 mL of saturated sodium chloride solution, dried over sodium sulfate and evaporated in vacuo. 3.7 g of the title compound are obtained in the form of a foam. The product has a PNMR spectrum Identical to the spectrum of the title compound prepared by the above Method A.

CS 268 156 B2

Example 36

Pivaloyloxymethyl-6 c * - (benzyloxycarbonylaininomethyl) penilcianate 1,1-dloxide

6.75 g (17 mmol) of the title compound a

3.34 ml (18.7 mmol) of N, Ν-diisopropylethylemine are dissolved in 50 ml of dimethylformamide, 2.72 ml (18.7 mmol) of chloroethyl pivalate are added and the mixture is stirred at room temperature for 20 hours. The reaction mixture is diluted with 300 ml of ethyl ether, washed twice with 100 ml of water each time, dried over sodium sulphate and evaporated in vacuo to an oily residue. The oily material was dissolved in ether (100 ml), washed with water (3.times.50 ml), dried (Na2 SO4) and concentrated in vacuo. 4.4 g of purified title product are obtained in the form of a viscous oil.

PNMR (deuterochloroform, tetramethylsilane, X-values):

1.20 (9H, a), 1.34 (ЗН, and)" 1.51 (3H, a), 3.64 <3H, m), 4.31 (1H, and), 4.60 (1H, d). 5.04 (2H, a), 5.71 (2H, q). 7.24 (5H, and). P ř í к 1 i 2 d 37

Pivaloyloxymethyl-6- (aminomethyl) -penicilate 1,1-dioxide salt with p-toluenesulfonic acid

1.8 g of the title compound (3.53 mmol) was hydrogenated in a mixture of 40 ml of tetrahydrofuran and 20 ml of water in the presence of 1.8 g of 10% palladium on carbon catalyst and in the presence of 1.77 g of 7.06 mmol. of pyridinium p-toluenesulfonate for 1.5 hours at 50 psi. The catalyst is filtered off with diatomaceous earth and tetrehydrofurene is distilled off from the filtrate in vacuo, 1.2 g of the title compound crystallizing, melting at 214 DEG-215 DEG C. with decomposition. PNMK (perdeuterodimethylsulfoxide, tetramethylsilane, hodnoty values);

1.16 (9H, s), 1.32 (3H, s), 1.48 (3H, s), 2.28 (3H, s), 3.34 (2H, m), 3.82 (1H) m), 4.60 (1H, a), 5.14 (1H, d, J 2 Hz), 5.75 (2H, ABq), 7.23 (4H, ABq).

For C ^ HH ^O ^N NS -C CH HSO SOH requires C, 48.16; H, 5.88; N, 5.11.

Found: C, 48.31; H, 6.11; N, 5.08.

Example 38 Chloromethyl-6- (D) - (2-azido-2-phenylacetemido) p-penicllenate

A solution of 12.0 g (0.03 mol) of sodium 6- [D- (2-azido-2-phenylacetamido) -penic acid in 25 ml of water was treated with 100 ml of methylene chloride and 10.17 g (0.03 mol). The mixture was adjusted to pH 7.5 with sodium bicarbonate, the organic layer was separated and the aqueous layer was extracted twice with 100 ml of methylene chloride each time. The combined organics were dried over sodium sulfate and the solvent was evaporated. The solid residue was triturated with 300 ml of ethyl acetate, the mixture was filtered, the filter cake was washed first with ethyl acetate and then with ethyl ether, and dried under nitrogen. 16.5 g (89%) of the tetrabutylemmonium salt are obtained.

12) 32 g (0.02 mol) of the above salt were mixed with 70 ml of chloroiodomethane and stirred at room temperature overnight. The reaction mixture is concentrated to dryness and the residue is purified by chromatography on 600 g of silica gel using a mixture of equal parts of ethyl acetate and hexane (v / v) as eluent. 8.1 g (95%) of the desired chloromethyl ester are obtained in the form of a pale yellow viscous oil.

PNMR (deuterochloroform values in ppm):

CS 268 156 B2

1.58 (з, ЗН), 1.68 (а, ЗН), 4.45 (s, W,

5.1 (α, 1 N), 5.5 - 5.9 (dd in, 4 N), 7.2 (d, 1 N), 7.4 (β, 5 N).

Example 39

Iodomethyl-6-CD- (2-azido-2-phenylaceteaido)] penicllanate

To a solution of 1.45 g (0.00342 mol) of the title compound in 30 ml of acetone was added nitrogen for 3 minutes, then 2.55 g (0.01714 mol) of sodium iodide was added and the resulting solution was stirred for 16 hours. The reaction mixture is clarified by filtration, the filtrate is concentrated in vacuo, the residue is taken up in 75 ml of chloroform and the solution is filtered. The chloroform filtrate was washed twice with 30 ml of saturated sodium chloride solution each time, dried over sodium sulfate and concentrated. This afforded 1.23% of the title compound as a foam.

PNMR (deuterochloroform, tetramethylsilane, ppm values):

1.53 (3H, a), 1.64 (ЗN, a), 4.37 (1H, s),

5.05 (1H, a), 5.56 (2H, m 4.11 Hz),

5.87 (2H, ABq), 7.31 (5H, s).

Example 40? - (Benzyloxycarbonylaminomethyl) -1,1-dioxopenicilanoyloxymethyl-6-? - (2-azido-2-phenylacetamido)} - penicillanate

0.56 g (1.43 mmol) of the title compound of Example 16 was dissolved in 50 ml of dichloromethane, added to 20 ml of water and adjusted to pH 8.6 with 1N sodium hydroxide. Sodium bicarbonate (0.121 g, 1.43 mmol) was added in portions, followed by tetrabutylammonium hydrogen sulphate (0.488 g, 1.43 mmol), maintaining the pH a with 1N sodium hydroxide in the range of 8.0 to 8.3 to almost at the end of the addition, when the pH was allowed to drop to 7.0. The reaction mixture was stirred for 15 minutes, then the layers were separated and the aqueous layer was extracted once with 30 mL of fresh dichloromethane. The organic layer was combined with the extract, dried over sodium sulfate and concentrated in vacuo to give a foamed 1,1-dioxide of tetrabutylammonium-Ctf (benzyloxycarbonylaminomethyl) penicillanate. This product was dissolved in 20 mL of acetone, added to a solution of 0.714 g (1.43 mmol) of the title compound in 15 mL of acetone, stirred at room temperature for 1 hour, and then concentrated in vacuo. The residue was stirred in 30 ml of ethyl acetate to give 0.42 g of crystalline tetrabutylammonium iodide. The filtrate was evaporated to give a 1.2 g of bovine residue which was chromatographed over 100 g of silica gel using 20% ethyl acetate in chloroform as eluent. Fractions of 20 ml are collected. Fractions containing pure product (Rf 0.22 by thin layer chromatography in the same solvent system) were combined and concentrated in vacuo. 0.61 g of purified title compound is obtained in the form of a foam.

PNMR (deuterochloroform, ppm values):

1.33 OH, s), 1.48 OH, s), 1.52 (ЗН, p), 1.59 (ЗН, в), 3.65 OK, n) 4.33 (1 H, s), 4.42 (1H,.), 4.61 (1H, W) , 5.05 (3H, 8) 5.53 (5H, ш), 7.24 (5H, «)» 7.32 (5H, s). P ř j [klad 41

N - (aminomethyl) -1,1-dioxopenicllanoyloxymethyl-6-CD-2-amino-2-phenylacetamido) -pentiliacanate-dipp-toluenesulfonate)

Prehydrogenate 2 g of 10% palladium on carbon in 20 ml of water, first add a solution of 0.96 g <1.226 mmol) of the title compound in 30 ml of tetrahydrofuran, and then 0.615 g (2.452 mmol) of pyridine / p-toluenesulfonate, and the mixture was hydrogenated at 50 psi for 1.5 hours. The catalyst was filtered through diatomaceous earth, which was washed with tetrahydrofuran and water. The filtrate is combined with the antifouling liquids and

Tetrahydrofurene was evaporated in vacuo. The aqueous residue was extracted three times with 30 ml of ethyl acetate each and then lyophilized to give 0.66 g of the title compound contaminated with the undiluted benzyloxycarbonyl compound. 1.0 g of fresh 10% palladium on carbon is prehydrogenated in 20 [mu] l of water, 0.5 g of the impure compound prepared above is dissolved in 30 ml of tetrahydrofuran and added to the pre-reduced catalyst slurry. Finally, 0.315 g of fresh pyridinium p-toluenesulfonate is added and the mixture is hydrogenated at 50 psi for 1.5 hours. The catalyst was separated and the purified title compound was isolated as described above. Yield 0.5 g.

PNMH (perdeuterodimethylsulfoxide, tetrsmethylsilane, 250 MHz, evaluation):

1.35 (6H, Wide ·), 1.47 (6H, 2.30 (6H, 3), 3.38 (2H, и) 3.94 (1H, m), 4.45 (1 H, »), 4.72 (1 H, with) 5.08 (1H, Broad., 5.31 (1H, broad) with) 5.45 (1H, d, J 4 Hx), 5.60 (1H, и), 5.93 (2H, a), 7.32 (ΘΗ, ABq), 7.48 (5H, and). Ex: t к 1 β d 42

1,1-dioxopenicilanoyloxymethyl-6- (benzyloxycarbonylaminomethyl) -penicillanate 1,1-dioxide

0.5 g (1.26 mmol) of the title compound of Example 16 is dissolved in 50 ml of dichloroethane, 10 ml of water are added and the pH is adjusted to 8.6 with 1N sodium hydroxide solution. After adding 0.106 g (1.26 mmol) of sodium bicarbonate first and then 0.428 g (1.26 mmol) of tetrabutylammonium hydrogen sulphate, the pH, which has fallen to 5.0, is adjusted to 7.5 with 1N sodium hydroxide, the mixture is stirred for 30 minutes After stirring at room temperature, the organic layer was separated, dried over sodium sulfate and concentrated in vacuo. The resultant tetrabutylammonium-6H- (benzyloxycarbonylaminomethyl) warbler 1,1-dloxide is dissolved in 20 ml of acetone. 0.47 g of iodomethyl-penicillanate (1,17-dioxide) prepared, for example, according to Godtfredsen et al., U.S. Pat. No. 4,342,772) in 15 ml of acetone is added, the mixture is stirred for 5 minutes and then concentrated in vacuo. The residue is stirred in 30 ml of ethyl acetate and 0.33 g of crystalline tetrabutylammonium iodide is isolated by filtration. .

The filtrate was concentrated in vacuo to give 0.82 g of the title compound as a foam.

PKMR (deuterochloroform, tetramethylsilane, values in ppm):

1.40 (3H, e), 1.42 (3H, s), 1.58 (6H, s), 3.41 (2H, m), 3.69 (3H, m), 4.40 (2H) , s), 4.58 (2H, from), 5.08 (2H, a),

5.59 (1H, m), 5.86 (2H, s), 7.29 (5H, s).

Example 1 a d 43 ·

1,1-dioxopenicllanoyloxymethyl-6 (t-eminomethyl) salt foamed with p-toluenesulfonic acid

1.2 g of 10% palladium on carbon are prehydrogenated in 10 ml of water, then 0.482 g (1.92 mmol) of pyridinium tosylate are added, followed by a solution of the title compound in 30 ml of tetrahydrofuran, and the resulting mixture is stirred for 1 hour. hydrogenates at a pressure of 50 psi. The catalyst was filtered off through diatomaceous earth, which was washed with tetrahydrofuran and water, the filtrate was combined with the washing liquid, and the tetrahydrofuran was removed by evaporation in vacuo. The aqueous residue was extracted once with 30 ml of ethyl acetate, the organic layer was dried over sodium sulfate and concentrated to give 60 mg of a solid residue. The aqueous layer was concentrated to 10 mL. Filtration gave 100 mg of the crystalline title compound, melting at 228-229 ° C with decomposition. PNMR (perdeuterodimethylsulfoxide, 250 MHz, ^ values):

1.35 (3H, s), 1.37 (3H, s), 1.49 (3H, s),

1.50 (3H, s), 2.29 (3H, s), 3.29 (1H,

CS 268 156 B2 dd, J = 1.7 »16.6 Hz), 3.39 (2H, o),

3.72 (1H, dd, J = 4.6, 16.6 Hz), 3.92 (1H, n), 4.60 (1H, s), 4.77 (1H, s), 5.21 (2H, α), 5.96 (2H, s), 7.31 (4H, ABq).

Analysis: calculated C, 44.17; H, 4.89; N, 6.18.

found 45.53% С »4.76% H, 6.10% N.

Lyophilization of the aqueous mother liquor afforded 160 mg of a solid which was suspended in a small amount of water, filtered, washed with a small amount of ethyl acetate and dried to give an additional 70 mg of purified title compound having the same melting point and PNMR spectrum as the above prepared product.

Example 44

Chloromethyl-бсС- (benzyloxycarbonylaminomethyl) foam

In analogy to Example 40, 0.396 g (1.0 mmol) of the title compound of Example 16 was converted to the tetrabutylemmonium salt. This salt was dissolved in 30 ml of bromochloromethane, stirred at room temperature for 18 hours, then concentrated in vacuo to a bovine residue and chromatographed on 50 g of silica gel using 20% ethyl acetate in chloroform as eluent. Fractions of 20 ml are collected. Fractions 6 to 10 are combined and concentrated in vacuo to a foamed product (0.25 g) which has Rf = 0.7 when thin layer chromatography in the same solvent system.

PNMR (deuterochloroform, tetrafluoromethylsilane, values (ppm in ppm):

1.37 (3H, 1.54 (3H, s), 3, 70 (3H, m) 4.38 (1H, with), 4.67 (1H, wide with) > 5.07 (2H, with), 5.66 (1H, d, J ³ 9 Hz), 5.70 (2H, ABq: 1, 7 ». 33 (5 H, s). P ř j í к 1 and d 45

Iodomethyl-6 <C - (benzyloxycarbonylamino ethyl) 1,1-dioxide foam

0.25 g (0.563 mmol) of the title product was dissolved in 15 mL of acetone and purged with nitrogen. After addition of 0.42 g (2.8 mmol) of sodium iodide, the resulting solution was stirred for 17 hours and then concentrated in vacuo. The solid residue was triturated with chloroform, the insolubles were filtered off and the filtrate was concentrated in vacuo to give 0.23 g of the title compound as a foam.

PNMR (deuterochloroform, tetramethylsilane, ppm):

1.39 (3H, s), 1.55 (3H, s), 3.64 (3H, tu), 4.23 (1H, s), 4.59 (1H, broad s), 5.04 (2H, s), 5.43 (1H, ra), 5.beta.3 (2H, ABq), 5.23 (5 H, s). Example 46

Methylene-bis [6 Λ - (benzyloxycarbonylaminomethyl) -1,1-dioxopenicilane]

0.17 g (0.429 mmol) of the title compound of Example 16 was converted to the tetrabutylammonium salt by the procedure of Example 40. This salt was dissolved in 10 mL of acetone and added to a solution of 0.23 g (0.429 mmol) of the product of the previous example in 10 mL of acetone. The mixture was stirred for 15 minutes, then concentrated in vacuo to a foamy residue which was stirred in 20 mL of ethyl acetate. Filtration of the suspension yielded 110 mg of tetrabutylemonlumide iodide in concentration of the filtrate in vacuo to give 0.28 g of the title compound as a foam.

PNMR (deuterochloroform, tetramethylsilane, J * values in ppm):

1, J6 (6H, a), 1.52 (6H, а), 3.73 (6H, η), 4.40 (2Η, в), 4.69 (2Н, wide в),

5.08 (4 N, N), 5.77 (4 N, N), 7.28 (4 N, N).

Р ř Í к 1. d 47 bla-β-toluenesulfonate) methylene-bla- [6 '- (aminomethyl) -1,1-dioxopenia targeted tu]

CS 268 I55 B2

The title compound ae was hydrogenated according to the procedure described in Example 43. After separation of the catalyst and evaporation of tetrahydrofuran, the aqueous residue was extracted with ethyl acetate (3 x 20 ml) and lyophilized. 0.19 g of the title product is obtained.

Claims (8)

A process for producing 1,1-dioxide of 6-aminoalkylpenic acid of the general formula I * or II * in which: R is hydrogen or methyl and n is 2 and represents the group -CH-- z. or a value of 1 and R ¹ represents a hydrogen atom, a moiety constituting an ether which is hydrolysable under physiological conditions, a 1,1-dloxopanallanoyloxymethyl group or a radical of formula (A) O CS 268 156 02 in them! j Y represents a hydrogen atom, a hydroxyl group, an alkenoyloxy group having from 2 to 7 carbon atoms, an alkoxycarbonyloxy group having from 2 to 7 carbon atoms, a benzoyloxy group or a benzoyloxy group with a monosubstituted C1-4 alkyl group, an alkoxycarbonyl group; and the pharmaceutically acceptable edible salts and acids thereof and the pharmaceutically acceptable cationic salts of the above compounds wherein R @ 1 is hydrogen, wherein the hydrogenolysis compound of formula III * or XV is hydrogenated in an inert solvent in the presence of a noble metal catalyst. * in which R is hydrogen or methyl and n is 2 and R ⁵ is -CH ₂ or n is 1 and R 8 is benzyl, the ester-forming moiety hydrolyzable from physiological conditions, 1,1-dloxopenitilanoyloxymethyl or the radical of formula VI in which Y * is hydrogen, benzyloxycarbonyloxy, C 2 -C 7 alkanoyloxy, C 2 -C 7 alkoxycarbonyloxy, benzoyloxy or benzoyloxy monosubstituted C 1 -C 4 alkyl, and C 1 -C 4 alkyl or C 1 -C 4 - ' Z is azidocappline or benzyloxycarbonylamino, and the resulting compound of formula (I *) or is optionally converted to its pharmaceutically acceptable acid addition salt or, when R 6 is hydrogen, to its pharmaceutically acceptable cationic salt. The method of item 1, wherein the corresponding starting materials of formula III * or IV * are used to form compounds of formula I * or II, * wherein R is as in 1, n has the value 1 в represents the moiety being an ester readily hydrolyzable under physiological conditions, selected from an zahrnující 2 3 2 3 'butyrolacton-4-yl moiety, and moieties of the formulas -CHR OCOR and -CHR OCOOR, wherein 2 3 R represents a hydrogen atom or a methyl group and R represents an alkyl group having 1 to 6 carbon atoms; CS 26Θ 156 B2 3. Method according to claim 2, characterized ее tlc That ее using the corresponding starting materials of formula XV III'nebo ₉ to give compounds of formula I * nsbo XI * wherein n is 1, R is not hydrogen and R · ^ represents a 1-ethoxycarbonyloxyethyl or pivaloyloxymethyl group; 4. A process according to claim 3, wherein the corresponding starting material of formula IV * _{f is used} to form a compound of formula XXf wherein R is as in 3 and R5 is pivaloyloxymethyl; 5. A process according to claim 1, wherein e is to use the corresponding starting materials of formula (XII) or (b) to form compounds of formula (1) and (b) XXf wherein n is 1, R is 1,1-dioxopenicilanoyloxymethyl and R is means hydrogen atom · 6. A method according to claim 5, wherein the corresponding starting material of the formula IV * is used to give the aluconines of the formula XI * in which n, R R and R have the meanings given in point 5. 7. The method of claim 1, wherein ea is to use the corresponding starting materials of formula III 'or IV * to form compounds of formula I * or II. * Wherein n is 2, R is -CH4- and R is hydrogen S «A method according to claim 7, characterized in that from using the corresponding starting material of Formula IV to afford a compound of formula XI * wherein n, R ¹ and R are defined as in point 7 · 9. A process according to claim 1, wherein the corresponding starting materials of the formula III * or IV * are used to form compounds of the formula X * or II *. where n has 1 * is 1, R is A and Y and R are hydrogen ·