CH633798A5 - Process for the preparation of ether derivatives of thienamycin - Google Patents

Description

The present invention relates to a method for producing ether derivatives of thienamycin, namely oxygen derivatives of the secondary alcohol group of antibiotic thienamycin.

Such derivatives are useful antibiotics.

The starting material for the preparation of the compounds obtainable according to the invention is thienamycin. The compound mentioned was described in U.S. Patent No. 3,950,357. This patent specifies the production and isolation of thienamycin. Thienamycin has the following structural formula:

sch2ch2nh2

cooh m

The compounds which can be prepared according to the invention have the following formula

SCH2CH, ÏÎH2 COOK

^ CCOR *

io in which R1 and R2 represent easily removable protective groups, correspondingly etherified, the protective groups removed, and that compounds obtained are converted, if appropriate, into the corresponding salts.

The compounds which can be prepared according to the invention can easily be represented by the following formula

20th

Th r or ■

-nh.

_COCM

ii A

25th

(II)

where R4 represents an ether residue.

The process according to the invention for the preparation of the ethers is characterized in that compounds of the formula are prepared.

In this formula, «Th» represents the bicyclic core of thienamycin and the hydroxy, amino and carboxyl groups of thienamycin are listed. M is hydrogen 30 or can also be a salt cation selected from the group of alkali or alkaline earth metals or an amine salt and R4 is an ether radical, R4 means alkyl, aryl, aralkyl and similar radicals, as are defined in more detail below that the group OR4 can be called ether 35.

There is an ongoing need for new antibiotics because, unfortunately, there is no static effectiveness for a known antibiotic due to its widespread use and resistant strains of pathogens have formed. In addition, the known antibiotics have the disadvantage that they are only effective against a certain type of microorganism. For this reason, the search for new antibiotics continues.

Unexpectedly, ether compounds 45 have now been found which have a broad antibiotic spectrum and can be used both in the treatment of animals and humans and for inanimate systems.

The object of the invention is to provide a method for producing a new class of antibiotics which have the basic nuclear structure of the antibiotic thienamycin but are characterized by the corresponding oxygen derivatives of thienamycin, namely the ethers. These antibiotics are highly effective in inhibiting the growth of a wide range of pathogens that are representative of both gram-positive bacteria, e.g. S. aureus, S. pyogenes and B. subtilis. The present invention also relates to the preparation of the non-toxic pharmaceutically acceptable salts of said Verbin-60 fertilizers.

R4 is preferably a substituted or unsubstituted straight-chain or branched alkyl radical having 1 to 6 carbon atoms, alkenyl or alkynyl each preferably having 2 to 6 carbon atoms, aryl, such as e.g. Phenyl, aralkyl, e.g. 65 benzyl, cycloalkyl with preferably 3 to 6 carbon atoms or a heteroaryl or heteroaralkyl radical (mono- or bicyclic), in which the alkyl part preferably has 1 to 3 carbon atoms and the heterocyclic ring ins

633798

has particular 4 to 10 atoms and the heteroatoms are selected from the group consisting of oxygen, nitrogen and sulfur; the groups mentioned above may be unsubstituted or substituted by radicals such as e.g. OH, SH, SR3 (R3 is lower alkyl or aryl such as phenyl), alkyl or alkoxy with 1 to 6 carbon atoms, halogen, e.g. Chlorine, bromine, fluorine and iodine, cyano, carboxy, sulfamino, carbamoyl, sulfonyl, azido, amino, substituted amino such as e.g. Alkylamino, including quaternary ammonium, wherein the alkyl radical has 1 to 6 carbon atoms, halo-alkyl, e.g. Trifluoromethyl, carboxyalkyl, carbamoyl-alkyl, nitrogen-substituted carbamoylalkyl, the aforementioned alkyl parts preferably having 1 to 6 carbon atoms in all radicals, amidino, guanidino, nitrogen-substituted guanidino, guanidino-lower alkyl and the like. Preferred examples of such ether radicals can be those in which R4 has the following meaning: benzyl, p-hydroxybenzyl, 4-amino-4-carboxybutyl, methyl, cyanomethyl, 2-pentenyl, β, β-diphenylethyl, methyldiphenylmethyl, triphenylmethyl, 2-methoxyphenyl, 2,6-dimethoxyphenyl, 2,4,6-trimethoxyphenyl, 3,5-dimethyl-4-isoxazolyl, 3-butyl-5-methyl-4-isoxazolyl, 5-methyl-3- phenyl-4-isoxazolyl, 3- (2-chlorophenyl) -5-methyl-4-isoxazolyl, 3- (2,6-dichlorophenyl) -5-methyl-4-isoxazolyl, D-4-amino-4-carbo- xybutyl, D-4-N-benzoylamino-4-carboxy-n-butyl, p-amino-benzyl, o-aminobenzyl, m-aminobenzyl, p-dimethylamino-benzyl, (3-pyridyl) methyl, 2-ethoxy l-naphthyl, 3-car-boxy-2-quinoxalinyl, 3- (2,6-dichlorophenyl) -5- (2-furyl) -4-iso-xazolyl, 3-phenyl-4-isoxazolyl, 5-methyl- 3- (4-guanidino-phenyl) -4-isoxazolyl, 4-guanidinomethylphenyl, 4-guanidi-nomethylbenzyl, 4-guanidinophenyl, o-sulfobenzyl, p-car-boxymethylbenzyl, p-carbamoylmethylbenzyl, m-fluorobenzyl, m-bromobenzyl, p-chlorobenzyl, p-methoxybenzyl,

1-naphthylmethyl, 3-isothiazolylmethyl, 4-isothiazolyl-methyl, 5-isothiazolylmethyl, guanylthiomethyl, 4-pyridyl-methyl, 5-isoxazolylmethyl, 4-methoxy-5-isoxazolylmethyl, 4-methyl-5-isoxazolylazolmethyl, 1-imyl 2-benzofuranylmethyl, 2-indolylmethyl, 2-phenylvinyl, 2-phenylethynyl, 1-aminocyclohexyl, 2- and 3-thienylaminomethyl,

2- (5-nitrofuranyl) vinyl, phenyl, o-methoxyphenyl,

0-chlorophenyl, o-phenylphenyl, p-aminomethylbenzyl,

1- (5-cyanotrizolyl) methyl, difluoromethyl, dichloromethyl, dibromomethyl, l- (3-methylimidazolyl) methyl, 2- or 3- (5-carboxymethylthienyl) methyl, 2 or 3- (4-carbamoylt-hienyl) -methyl, 2- or 3- (5-methylthienyl) methyl, 2- or

3- (methoxythienyl) methyl, 2- or 3- (4-chlorothienyl) methyl, 2- or 3- (5-sulfothienyl) methyl, 2- or 3- (5-car-boxythienyl) methyl, 3 - (l, 2,5-thiadiazolyl) methyl, 3- (4-methoxy-l, 2,5-thiadiazolyl) methyl, 2-furylmethyl, 2- (5-nitrofuryl) methyl, 3-furylmethyl, 2 -Thienylmethyl, 3-thienylmethyl, tetrazolylmethyl, benzamidinomethyl and cyclohexylamidinomethyl.

The ether group can also represent a radical of the following formula:

- (CH2) "ZR"

where n is 1-4, Z is oxygen, sulfur, carbonyl or nitrogen X and R "has the following meaning: hydrogen, amino, mercapto, hydroxy, alkylamino, dialkylamino, alkyl, alkylthio, anylthino, alkoxy, anylony, alkenyl, alkynyl , Aryl, aralkyl, acylcycloalkyl, heteroaryl or heteroaralkyl.

The ether group can also mean a radical of the following formula:

-CHR '"R" "

wherein R '"denotes hydrogen, amino, mercapto, hydroxy, alkylamino, dialkylamino, alkyl, alkylthio, arylthio, alkoxy, aryloxy, alkenyl, alkynyl, aryl, aralkyl, cycloalkyl, heteroaryl or heteroaralkyl and R" "denotes amino, hydroxy, azido, Carbamoyl, guanidino, acyloxy, halogen, sulfamino, tetrazolyl, sulfo, carboxy, carbalkoxy, phosphono, alkoxy or akylthio.

Examples of this group are:

a-aminobenzyl, a-amino- (2-thienyl) -methyl, a- (methyl-mino) -benzyl, a-amino-methylmercaptopropyl, a-amino-3-or 4-chlorobenzyl, a-amino-3- or 4-hydroxybenzyl, a-amino-2,4-dichlorobenzyl, a-amino-3,4-dichlorobenzyl, D (-) - a-hydroxybenzyl, a-carboxybenzyl, a-amino- (3-thienyl) methyl, D - (-) - a-Amino-3-chloro-4-hydroxybenzyl, a-amino- (cyclohexyl) -methyl, a- (5-tetrazolyl) -benzyl, 2-thienyl-carboxymethyl, 3-thienyl-carboxymethyl,

2-furylcarboxymethyl, 3-furylcarboxymethyl, a-sulfamino-benzyl, 3-thienyl-sulfaminomethyl, a- (N-methylsulfamino) -benzyl, D (-) - 2-thienylguanidinomethyl, D (-) - a-guanidino-benzyl, a-guanylureidobenzyl, a-hydroxybenzyl, a-azido-benzyl, a-fluorobenzyl, 4- (5-methoxy-l, 3-oxadiazolyl) ami-nomethyl, 4- (5-methoxy-1,3-oxadiazolyl) - hydroxymethyl, 4- (5-methoxy-l, 3-sulfadiazolyl) hydroxymethyl, 4- (5-chlorothienyl) aminomethyl, 2- (5-chlorothienyl) hydroxymethyl, 2- (5-chlorothienyl) carboxy- methyl, 3- (1,2-thiazolyl) aminomethyl, 3- (1,2-thiazolyl) hydroxymethyl,

3- (1,2-thiazolyl) carboxymethyl, 2- (1,4-thiazolyl) amino-methyl, 2- (1,4-thiazolyl) hydroxymethyl, 2- (1,4-thiazolyl) carboxymethyl , 2-benzothienyl-aminomethyl, 2-benzothienyl-hydroxymethyl, 2-benzothienyl-carboxymethyl, a-sulfo-benzyl, a-phosphonobenzyl, a-diethyl-phosphono and a-monoethylphosphono.

A particularly preferred class of ether residues are terminally substituted groups in which the substituent is a basic residue which is substituted or unsubstituted, e.g. Amino, amidino, guanidino, guanyl and nitrogen-containing mono- and bicyclic heterocycles (aromatic and non-aromatic), in which the heteroatom or the heteroatoms can also be oxygen and sulfur in addition to the nitrogen. Such preferred substituted ether residues can be represented by the following formula.

- (CH2) m-A- (CH2) n-Y

wherein m and n are 0 or an integer from 1 to 5, A is oxygen, NR '(R' is hydrogen or a lower alkyl radical having 1 to 6 carbon atoms) or sulfur or A is a simple bond, and Y has one of the meanings below can:

1.) Amino or substituted amino:

©

-N (R) 2 and -N (R) 3

wherein the values for R are independently selected from the following group: hydrogen; N (R ') 2 (R' is hydrogen or lower alkyl of 1 to 6 carbon atoms); Lower alkyl and lower alkoxy of 1 to 6 carbon atoms; Lower alkoxy lower alkyl, wherein the alkoxy part has 1 to 6 carbon atoms and the alkyl part has 2 to 6 carbon atoms; Cycloalkyl and cycloalkylalkyl, wherein the cycloalkyl ring has 3 to 6 carbon atoms and the alkyl part has 1 to 3 carbon atoms; two groups R together with the nitrogen atom to which they are attached can form a ring with 3 to 6 atoms.

4th

s io

15

20th

25th

30th

35

40

45

50

55

60

65

5

633798

2.) Amidino and substituted amidino:

-N = C-N (R) 2

I.

R s wherein the value for the substituent R is independently selected from the following group: hydrogen; N (R ') i (R' is hydrogen or lower alkyl of 1 to 6 carbon atoms); Lower alkyl or lower alkoxy, each with 1 to 6 carbon atoms; Lower alkoxy lower alkyl, in which the alkoxy part has 1 to 6 carbon atoms and the alkyl part has 2 to 6 carbon atoms (if the lower alkoxy lower alkyl radical is bonded to the carbon atom, the alkyl part has 1 to 6 carbon atoms); Cycloalkyl and cycloalkyl-is alkyl, wherein the alkyl portion has 1 to 3 carbon atoms; two groups R together with the atoms to which they are attached can form a ring with 3 to 6 atoms.

/ ^ i w

3.) Guanidino and substituted guanidino: -NH-C-N (R) 2

II

NO

wherein the substituent R is defined above under 2.).

4.) Guanyl and substituted guanyl:

-C = NO

I.

NO. 2

wherein the substituent R is defined above under 2.).

5.) Nitrogen-containing mono- and bicyclic heterocycles (aromatic and non-aromatic) with 4 to 10 core atoms, in which the heteroatom or the heteroatoms can also be oxygen or sulfur in addition to nitrogen. Such heterocycles, which are preferred, are listed below (R 'is hydrogen or lower alkyl having 1 to 6 carbon atoms:

N (R ') £ 2

• R '

20th

25th

â \.

\\ ./

■ O

35

40 The ethers which can be prepared according to the invention are encompassed by structural formula II given above, and preferred ether derivatives of thienamycin are those which have a relatively low molecular weight and are also hydrophilic. For example, in relation to the ethers obtainable according to the invention, the following radicals are preferred and representative: methoxymethyl, hydroxymethyl, methoxyethyl, dimethylaminomethyl, dimethylaminoethyl, methylthioethyl, amidinoethyl, guanidinoethyl and the like.

In general, the antibiotics obtainable according to the invention are prepared from thienamycin starting compounds of the formula III in which all residues are protected by protective groups:

f o '

_jsch2ch2nr1r2

-n-

.COCR-

iii

"OR TMR1R2

-COOR * purple r-OR4

Th-h-NH2 ICOGH

IIa

In these compounds, R1 and R2 are common nitrogen protecting groups (usually R1 or R2 mean hydrogen) and R3 is a well known carboxyl protecting group. The preparation of the compounds of the formula III which contain protective groups on the nitrogen atom and on the acid group is described in detail below and also in the preparations. The only limitation for protection groups R1, R2 and R3 is that

633 798

6

they do not adversely affect the desired reaction and that they are ultimately easily removable to produce compounds of the formula I. The definitions of "protecting groups" used here are the same as those described in detail in U.S. Patent No. 3,697,515.

The preferred nitrogen protecting groups in the starting material of formula III are in particular: carbobenzyloxy, ring-substituted carbobenzyloxy, such as e.g. o- and p-nitrocarbobenzyloxy, p-methoxycarbobenzyloxy, chloroacetyl, bromoacetyl, phenylacetyl, t-butoxycarbonyl, tri-fluoroacetyl, bromoethoxycarbonyl, 9-fluorenylmethoxycarbonyl, dichloroacetyl, o-nitrophenylsulfenyl, 2,2,2-trichcarbonloro , Bromo-t-butoxycarbonyl, phenoxyacetyl; Protecting groups for other than acyl groups are e.g. Alkylidene such as benzylidene, salicylidene and the like.

Preferred examples of suitable blocking esters R3 for the starting materials of the formula III are listed below:

(i) R3 = CRaRbRc wherein at least one of the substituents Ra, Rb and Rc represents an electron donor, e.g. p-methoxyphenyl, 2,4,6-trimethylphenyl, 9-anthryl, methoxy, CH2SCH3, tetrahydrofur-2-yl, tetrahydropyran-2-yl or fur-2-yl. The remaining substituents Ra, Rb and Rc can be hydrogen or an organic substituent. Suitable ester groups of this type are e.g. p-methoxybenzyloxycarbonyl and 2,4,6-trimethylbenzyloxycarbonyl.

(ii) R3 = CRaRbRc, wherein at least one of the substituents Ra, Rb and Rc represents an electron attractive group, e.g. Benzene, p-nitrophenyl, 4-pyridyl, trichloromethyl, tribromomethyl, iodomethyl, cyanomethyl, ethoxycarbonylmethyl, arylsulphonylmethyl, 2-dimethylsulfoniumethyl, o-nitrophenyl or cyano. Suitable esters of this type include benzoylmethoxycarbonyl, p-nitrobenzyloxycarbonyl, 4-pyridylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl and 2,2,2-tribromoethoxycarbonyl.

(iii) R3 = CR'RbRc, wherein at least two of the substituents Ra, Rb and Rc represent a hydrocarbon radical, such as alkyl, e.g. Methyl or ethyl or aryl, e.g.

Phenyl, and the remaining substituent Ra, Rb and Rc, if present, is hydrogen. Suitable esters of this type include e.g. t-butyloxycarbonyl, t-amyloxycarbonyl, diphenylmethoxycarbonyl and triphenylmethoxycarbonyl.

(iv) R3 = Rd, where Rd is adamantyl, 2-benzyloxyphenyl, 4-methylthiophenyl or tetrahydropyran-2-yl.

Silyl esters belonging to this category of protective groups can usually be prepared from a halosilane or a silazane of the following formula R43SIX R4aSIX R43Si.NR42; R43Si.NH.COR4; r43Sì.NH.CO.NH.SìR43; R4NH.CO.NR4. SiR4s; or R4C (OSiR43); HN (SiR43) 2 wherein X 'represents a halogen atom, such as chlorine or bromine, and the various R4 radicals, which are the same or different, represent hydrogen or alkyl, such as e.g. Methyl, ethyl, n-propyl, isopropyl, aryl such as phenyl or aralkyl, e.g. Benzyl residues.

In this context, it is found that preferred protecting groups R3 include those groups which fall under the definitions given above, such as e.g. Aralkyl, haloalkyl, alkanoyloxyalkyl, alkoxyalkyl, alkenyl, substituted alkyl or aralkoxyalkyl as well

Alkylsilyl, wherein alkyl has 1 to 10 carbon atoms. For example, preferred «blocking groups» R3 benzyl, phenacyl, p-nitrobenzyl, methoxymethyl, trichloroethyl. Trimethylsilyl, tributyltin, p-methoxybenzyl or Benz-5 hydryl. These protecting groups are generally known to be easily removed from cephalosporin and penicillin compounds.

In general, the compounds of the formula III are first reacted with N-acylating agents in order to protect the nitrogen-containing group and only then is the protective group for the carboxylic acid radical introduced; however, it is also possible to introduce the substituents R '/ R2 and R3 simultaneously by a common reaction, if possible. The removal of the protective groups is preferably carried out in a single step which is generally well known, but depending on the nature of the substituents R '/ R2 and R3, the protective groups can also be removed by carrying out several steps.

The introduction of the desired protective groups can be carried out by reacting the free thienamycin of the formula I accordingly or by using an already protected derivative of thienamycin, e.g. Tris-silylate-thienamycin

25th

Th-

30th

-OTMS

-nhtms converts i-co2tms, in which TMS represents a triorganosilyl radical, such as e.g. Trimethylsilyl; the preparation of such compounds is described in detail below.

The preferred nitrogen protecting groups R1 and R2 are those in which R1 is hydrogen and R2 is a substituted or an unsubstituted carbobenzyloxy radical of the formula

40 - ^ 2 R =

-OCH,

45 means where n is 0, 1 or 2 (when n is 0, R6 is hydrogen) and R6 is lower alkoxy or nitro. Another particularly preferred nitrogen protecting group is the bromo-t-butoxycarbonyl radical.

A preferred acid protecting group R3 is the benzyl radical or substituted benzyl groups, e.g. p-nitrobenzyl:

TT = -CH.

^ (R «) n

55

where n is 0.1 or 2 (n is 0 when R 'is hydrogen) and R6 is lower alkoxy or nitro.

A preferred starting material of formula III can be indicated by the following formula:

60

Th

65

raw

tNHR ^

co "c

C02CH2

(R6) n

(m)

7

633798

2 9

R = -C-O-CH

i2 ^ 0 <

r2 =

u CH3

C-0-Ó-CK-.E2 ÓH, 2

(R ') n, or

In general, a compound of formula III can be prepared by reacting thienamycin in water or in a mixture of water and a polar organic solvent, e.g. Oxane, treated with an acyl halide in the presence of a base, e.g. Carbobenzyloxy chloride. The N-acyl-thienamycin derivative obtained can be isolated by known methods. The acid protecting group is usually introduced in a second step, reacting the N-blocked thienamycin with an activated halide, e.g. Benzyl bromide, in a solvent such as hexamethyl phosphoramide or the like in a temperature range from 0 to 60 ° C, the reaction may take a few minutes to 4 hours. The compounds of the formula III obtained which have both N and acid protecting groups are then converted into the intermediate compounds of the formula purple; this procedure will be described later. The protective groups in the intermediate compounds of the formula purple are subsequently removed, the compounds of the formula II according to the invention being obtained by well-known processes, including hydrolysis and hydrogenation. When using preferred N and acid protecting groups, the preferred method for removing the protecting groups is based on hydrogenation; the intermediate compounds of the formula lilac, which are in a solvent such as a alkanol, is hydrogenated in the presence of a hydrogenation catalyst, e.g. Platinum or palladium or their oxides.

When preparing compounds of the formula II, the N-protected compounds of the following formula (1) can first be prepared as follows:

Th.

raw

, nh2 Lcooh raw

Th--

1 2 ■ no r

Lcooh

1

/ V

In general, the N-protected compounds of formula 1 above, wherein R1 and R2 are hydrogen or acyl, are prepared by reacting thienamycin of formula I with an acylating agent, e.g. an acyl halide or acid anhydride, such as aliphatic, aromatic, heterocyclic, araliphatic or heterocyclic aliphatic carboxylic acid halides or the corresponding anhydrides. Other acylating agents can also be used, e.g. mixed carboxylic acid anhydrides and especially lower alkyl esters of mixed carboxylic carboxylic acid anhydrides; carbocyclic acids can also be used in the presence of carbodiimide, e.g. 1,3-dicyclohexylcarbodiimide, and an activated ester, and an activated ester of a carboxylic acid, e.g. p-nitrophenyl ester. The acylation is usually carried out in a temperature range from approximately -20 ° C. to approximately 100 ° C., in particular in a range from −8 ° C. to 25 ° C. You can use any solvent in which the

35

40

45

50

Starting materials are soluble and are inert to the reaction, e.g. polar solvents such as water, alcohols and polar organic solvents generally such as methylformamide (DMF), hexamethylphophoramide (HMPA), acetone such as oxane, tetrahydrofuran (THF), acetonitrile, heterocyclic amines such as e.g. Pyridine, ethyl acetate, aqueous mixtures of the solvents mentioned and also halogenated solvents, such as Methylene chloride and chloroform, the reaction may take from about 5 minutes to a maximum of 3 hours, but generally a reaction time of from about half an hour to an hour is sufficient. The following equation shows this procedure using a carboxylic acid halide; it is to be understood, of course, that the halide mentioned can be replaced by a carboxylic anhydride or by another functionally equivalent acylating agent to give the same product.

OH

ch2ch2nh2 cooh

Acyl halide ch2ch2nr1r2

cooh

In general, if the acylating agents mentioned above are used, namely acid halides (suitable halides are derived from chlorine, iodine or bromine) or correspond to the anhydrides, the reaction is carried out in water or an aqueous mixture of a polar organic solvent, e.g. Acetone, dioxane, THF, DMF, acetonitrile or the like in the presence of a suitable base acceptor such as e.g. NaHCOs, MgO, NaOH, K2HPO4 and the like.

60 In the event that the acylating agent is sensitive to water, it is sometimes advantageous to carry out the acylation in a non-aqueous solvent. Triorganosilyl or tin derivatives of thienamycin are often suitable for this purpose. The silylation of thienamycin proceeds quickly and tris-triorganosilyl derivatives, e.g. Tris-trimethylsilyl-thiena-mycin Th (TMS) 3:

633798

Th

■ OTMS -NHTMS uCOOTMS

Such derivatives, which are readily soluble in organic solvents, are expediently reacted by reacting thienamycin with an excess of hexamethyl disilazane and a stoichiometric amount of tri-methylchlorosilane at a temperature of 25 ° C., stirring vigorously in a nitrogen atmosphere. The resulting ammonium chloride is then removed by centrifugation, and the solvent is removed by evaporation to give the desired silyl derivative. In general, the starting material of the formula III can be prepared from nitrogen-protected compounds of the formula L according to the following reaction scheme: 20

Th r oh

1 2

-NR R

l cooh

->

Th

■ coor-

2.) Reaction of an alkali metal salt of a compound of formula X with an activated alkyl halide, e.g. Methyl iodide, benzyl bromide or m-phenoxybenzyl bromide, p-t-butylbenzyl bromide, pivaloyloxymethyl chloride and the like. Suitable reaction conditions include solvents such as e.g. Hexamethylphosphoramid and the like, and one works in a temperature range from 0 to 60 ° C for a few minutes up to 4 hours.

3.) reaction of the compound of formula JL with an alcohol, such as e.g. Methanol, ethanol, benzyl alcohol and the like. The reaction can be carried out in the presence of a carbodimide as a condensing agent, e.g. Cyclohexylcarbodiimide and the like. Suitable solvents include CHCb, CH3CH, CH2CI2 and the like and one works at temperatures from 0 ° C to the reflux temperature for 15 minutes up to 18 hours.

rOK

1 2 ■ NR R

25th

1

III

wherein all substituents are defined above. 30th

Usually the conversion of 1 - III is carried out according to known methods. Such methods include the following methods:

1.) Reaction of a compound of formula JL with a 35 diazoalkane, e.g. Diazomethane, phenyldiazomethane, diphenyldiazomethane and the like in a solvent such as e.g. Dioxane, ethyl acetate, acetonitrile and the like at a temperature from 0 to the reflux temperature of 2 minutes to 2 hours. 40

4.) Reaction of a nitrogen acylated acid anhydride of L prepared by the reaction of the free acid of formula III with an acid chloride, e.g. Ethyl chloroformate, benzyl chloroformate and the like, with an alcohol as specified under 3.), using the same reaction conditions as mentioned under 3.). The anhydride is made by combining a compound of formula III with the acid chloride in a solvent such as e.g. Tetrahydrofuran (THF), CH2CI2 and the like at a temperature from 25 ° C to the reflux temperature for 15 minutes to 10 hours.

5.) Implementation of labile esters of thienamycin of formula (I) such as Trimethylsilyl ester, dimethyl-t-butylsilyl ester or the like with RX ', where X' is halogen, e.g. Bromine or chlorine and R is defined above in a solvent such as e.g. THF, CH2CI2 and the like at a temperature from 0 ° C to the reflux temperature for 15 minutes to 16 hours. The implementation can e.g. proceed according to the following scheme:

Th-

• OTMS

N-acylation -NHTMS - * Th-

rOTMS JTOTMS

i. "2 _, -_ esterification, j 2 ■ NR TMS -> Th-f-NR TMS

lcootm3

m: ootms cooh *

mild hydrolysis ^

Th

■ oh nhr2

Lcoor3

wherein TMS is a triorganosilyl group, e.g. Trimethylsilyl and the other substituents are already defined above.

The reaction schemes listed above are well known for the related bicyclic antibiotic β-lactams, and in all organic syntheses to be carried out there are generally no critical reaction parameters in the preparation of N-acyl- and carboxylthie-namycin (starting materials of the formula III).

In general, the compounds which can be prepared according to the invention are prepared using well-known etherification

ions (III-III a) on the secondary alcohol group of the thienamycin of the formula III, which has protective groups

60 points performed. (The removal of the protecting groups (Illa — II) then gives the compounds mentioned, which were discussed above.) Such methods (III — purple) include:

ss 1.) For the preparation of the corresponding ether, an acid-catalyzed reaction of compounds of the formula III in a diazoalkane, such as e.g. Diazomethane, phenyldiazomethane, diphenyldiazome

9

633798

than and the like in an inert solvent such as e.g. Dioxane, tetrahydrofuran (THF), halogenated hydrocarbons, e.g. CH2CI2, ethyl acetate and the like in the presence of a catalytic amount of a strong acid or a Lewis acid, e.g. Toluene sulfonic acid, trifluoroacetic acid, fluoroboric acid, boron trifluoride and the like, at a temperature of -78 ° to 25 ° C for a few minutes up to 2 hours.

2.) To prepare the corresponding ether mentioned, the reaction of compounds of formula III is preferably carried out with an alkylating agent, such as e.g. an active halide such as methyl iodide, benzyl bromide, m-phenoxybenzyl bromide and the like, alkyl sulphonates such as e.g. Dimethyl sulfate, diethyl sulfate, methyl fluorosulfonate and the like in the presence of a strong base capable of forming the alcoholate of a strong base capable of forming the alcoholate anion of a compound of Formula III. Suitable bases include alkali and alkaline earth metal oxides and hydroxides, alkali metal alkoxides such as e.g. Potassium tertiary butoxide, tertiary amines such as e.g. Triethylamine, alkali metal alkyl and aryl compounds such as e.g. Phenyllithium, as well as alkali metal amides, e.g. Sodium amide. Suitable solvents are any inert anhydrous solvent such as e.g. t-butanol, dimethylformamide (DMF), THF, hexamethylphosphoramide, (HMPA), dioxane and the like, and the procedure is carried out at a temperature of -78 to 25 ° C for a few minutes to 4 hours.

The compounds of the formula II obtained according to the invention can form a wide variety of pharmacologically acceptable salts with inorganic and also organic bases; these include e.g. Metal salts derived from alkali metal or alkali earth metal hydroxides, carbonates or bicarbonates and salts derived from primary, secondary or tertiary amines, e.g. Monoalkylamines, dialkylamines, trialkylamines, lower alkanolamines, di-lower alkanolamines, lower alkylenediamines, N, N-diaralkyl- or -alkylenediamines, aralkylamines, amino-substituted lower alkanols, lower alkanols substituted with N, N-di-lower alkylamino, with amino-, polyamino- and guanidino-substituted lower alkanoic acids and also nitrogen-containing heterocyclic amines. Representative examples are salts derived from sodium hydroxide, sodium carbonate, sodium bicarbonate, potassium carbonate, potassium hydroxide, calcium carbonate, trimethylamine, triethylamine, piperidine, morpholine, quinine, lysine, protamine, arginine, procain, ethanolamine, morphine, benzylamine, ethylenediamine Derive N, N'-dibenzyl-ethylenediamine, diethanolamine, piperazine, dimethylamino-ethanol, 2-amino-2-methyl-1-propanol, theophylline, N-methylglucamine and the like. Acid addition salts can also be prepared which are also pharmacologically active, e.g. Salts with hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, toluene-p-sulfonic acid and methanesulfonic acid.

The salts can be mono salts, e.g. the mononetrium salt, which is generally obtained by reacting 1 equivalent of sodium hydroxide with one equivalent of the compound of the formula II. However, mixed salts can also be obtained by treating 1 equivalent of a mono salt with 1 equivalent of a different base. However, there is also the possibility of preparing salts by adding 1 equivalent of a base containing a divalent cation, e.g. Calcium hydroxide, to be treated with one equivalent of the product of formula II. In addition, mixed salts and esters which could be obtained by reacting the compound of formula II with one equivalent of sodium hydroxide and then with one equivalent of lactic acid fall within the scope of the present invention. The salts obtainable according to the invention are all pharmacologically acceptable, non-toxic derivatives which can be used as active additives in suitable unit dosages of a pharmaceutical type. The compounds obtainable according to the invention can also be administered together with other active compounds in order to obtain a further spectrum of activity. In addition, the present salts and also the corresponding ester and amide derivatives can serve as intermediates in the preparation of the carboxylic acid compound of the formula II mentioned above.

The thienamycin derivatives obtained according to the invention are valuable antimicrobial substances which are different from a wide variety of gram-positive and gram-negative pathogens, such as e.g. Bacillus subsilis, Salmonella schottmuelleri and Proteus vulgaris, behave actively. For example, the free acid and especially its salts, e.g. Amine and metal salts, preferably the alkali metal and alkaline earth metal salts, useful bactericides and they can be used to remove sensitive pathogens from dental and medical devices, for the separation of microorganisms and for therapeutic use in humans and animals. For the latter purpose, it is possible to use pharmacologically acceptable salts with inorganic and organic bases, as are generally known for the administration of penicillins and cephalosporins. So e.g. salts of alkali metals and alkaline earth metals are preferably used and also salts of primary, secondary and tertiary amines. These salts can be combined with pharmaceutically acceptable liquid and solid carriers into suitable dosage units in the form of pills, tablets, suppositories, syrups, elixirs and the like, all of which are prepared by generally known methods.

The new compounds obtained according to the invention are valuable antibiotics which are active towards a wide variety of gram-positive and gram-negative bacteria and can therefore be used in human therapy and also in animals. The compounds mentioned can therefore be used as antibacterial drugs for the treatment of infections caused by gram-positive or gram-negative bacteria, such as e.g. against Staphyloccus aureus, Escherichia coli, Klebsiella pneumoniae, Bacillus subtilis, Salmonella typhosa, Pseudomonas and Bacterium proteus. The antibacterial agents obtained according to the invention can also be used as additives for animal feed, as preservatives for food and as disinfectants. For example, be used in aqueous agents in concentrations of 0.1 to 100 parts of antibiotic per million parts of solution to destroy or inhibit the growth of harmful bacteria on medical and dental equipment; furthermore such agents can be used as bactericides for industrial applications, e.g. in water-based paints or in the screen washer of paper mills, to suppress the growth of harmful bacteria.

The products obtained according to the invention can be used alone or in combination with one or more other active ingredients in pharmaceutical preparations. These antibiotics can be used in the form of capsules, tablets, powders, in liquid solutions or in the form of suspensions or elixirs. They can be administered orally, intravenously or intramuscularly.

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The pharmaceutical preparations are preferably in such a form that they are suitable for absorption in the gastrointestinal system. Tablets and capsules for oral administration may be in unit dosage form and may contain conventional excipients such as e.g. Binders e.g. Syrup, acacia resin, gelatin, sorbitol, tra-gacanth resin or polyvinylpyrrolidone; Fillers such as Lactose, sugar, corn starch, calcium phosphate, sorbitol or glycerin; Lubricants such as Magnesium stearate, talc, polyethylene glycol and silica; decay accelerating agents such as Potato starch; or also acceptable wetting agents, e.g. Sodium lauryl sulfate. The tablets can be coated in a manner known per se. Oral liquid preparations may be in the form of aqueous suspensions, oil suspensions, solutions, emulsions, syrups, elixirs, etc., or may be provided as a dry product for mixing with water or other suitable carriers before use. Such liquid preparations can be conventional additives such as e.g. Suspending agents such as sorbitol syrup, methyl cellulose, glucose sugar syrup, gelatin, hydroxyethyl cellulose, car-boxymethyl cellulose, aluminum stearate gel or hydrogenated edible fats, e.g. Contain almond oil, fractionated coconut oil, oily esters, propylene glycol or ethyl alcohol; further they can contain preservatives, e.g. Methyl or propyl p-hydroxybenzoate or sorbic acid. Suppositories usually contain common suppository bases, such as Cocoa butter or other glycerides.

Means for injections can be provided in unit dosage form in ampoules or in containers containing several doses with the addition of a preservative. The agents can be present as suspensions, solutions, emulsions in oily or aqueous carrier materials and they can contain formulation agents, such as suspending, stabilizing and / or dispersing agents. The active ingredient may also be in the form of a powder which may be coated with a suitable carrier, e.g. sterile, pyrogen-free water, is turned on before use.

The compositions can also be prepared in a form suitable for absorption through the mucous membranes of the nose or throat or the bronchial tissue, as a rule being provided as powdered or liquid spray or inhalation agents, pastilles, neck brush compositions, etc. For the treatment of eyes and ears, the agents can be provided in the form of individual capsules, in liquid or semi-solid form or drops, etc. can also be used. For local use, they can be formulated in hydrophobic or hydrophilic raw materials as ointments, creams, solutions, colors, powders, etc.

In addition to a carrier, the compounds obtained according to the invention can contain other constituents, such as stabilizers, binders, antioxidants, preservatives, lubricants, suspending agents, thickeners or flavoring agents and the like. In addition, other active ingredients may also be present in the agents to achieve a broader spectrum of antibiotic activity.

In veterinary medicine, e.g. as preparations to be injected into the breast are manufactured either with long-term effects or with quick release. The dosage to be administered depends largely on the condition and weight of the animal to be treated, the nature of the infection, the route and the frequency of administration, with parenteral administration for general infections and oral administration for intestinal infections being preferred. In general, a daily oral dosage amount is about 15 to 600 mg of the active ingredient per kg of body weight, preferably dividing the doses and administered one or more times a day. A preferred daily dosage amount for adult humans is in the range of about 80 to 120 mg of the active ingredient per kg of body weight.

The present preparations can be used in various unit dosage forms, e.g. in solid or liquid oral digestible dosage form. The preparations per unit of dosage, whether solid or liquid, can contain 0.1% to 99% of the active material, with a range of about 10 to 60% being preferred. The preparations will generally have from about 15 mg to about 1500 mg of the active ingredient, although it is generally preferred to use a dosage amount that is in the range of about 250 mg to 1000 mg. In parenteral administration, the 20 unit dose is usually the pure compound in a slightly acidified sterile water solution or in the form of a soluble powder intended for a solution.

In the examples below, preferred embodiments of the method according to the invention are described. As already mentioned above, the compounds which can be prepared according to the invention are represented by the following formula

30th

Th r

_NH2 L-COOH

in which formula the three functional groups are given.

40

IT

4S Th -hîH-C-ÇH

50

LcOONa

Preparation of the sodium salt of N- (0-formyl) -D-mandeloylthienamycin

124 mg of sodium bicarbonate, 8 ml of dioxane are added in succession to 40 mg of thienamycin in 10 ml of water at a temperature of 0 ° C. Then 1.2 equivalents of N- (O-formyl) are added to this mixture for one minute. -1 -mandeloyl chloride stirred. After 6 minutes of the total reaction time, the mixture obtained is extracted three times with ethyl ether which is so cold. Electrophoresis of an aqueous portion (0.05M, pH 7, aqueous phosphate buffer, 50 V / cm., 20 minutes shows 67% preservation to the desired product. Then the pH is adjusted by adding a 1 molar phosphoric acid solution is set to 2.2 and the solution is extracted three times with ethyl acetate, the ethyl acetate solution (EtO Ac) is dried over magnesium sulfate and extracted twice with 2 equivalents of a sodium bicarbonate solution

11

633798

after lyophilization contains 164 optical density units (ODU) at 302 nm by UV analysis at pH 7.0, 95% of which are extinguished after one hour after treatment with hydroxylamine. The yield is 53%. As before, electrophoresis shows a spot by bioautography, 4 cm in the direction of the anode.

NMR (8D2O) 1.30 (d, J = 6Hz, CHsCH); 2.8-3.7 (m, CH2), 4.0-4.5 (m, CH β-lactam), 4.73, HDO; 5.97 (s, CsHsCHOCHO), 7.53 (s, CaHs), 8.30 (s, CaHsCHOCHO).

Preparation 2

"OH H

Th 4-NC-CH-

11 1

O OH L-COONa

Preparation of the sodium salt of N-D-Mandeloylthie-namycin

The compound mentioned in the title is prepared by the method of preparation 1, but before the extraction with ethyl acetate, the aqueous extract is left to stand at a temperature of 25 ° C. for one hour. Electrophoresis (50 V / cm., 20 min., PH 7, aqueous phosphate, 0.05 M) shows a spot by bioautography, 4 cm towards the anode.

NMR (5D20) 1.50 (d, J = 6 Hz, CHaCH; 2.8-3.8 (m, CH2), 4.2-4.6 (m, CH ß-lactam); 4.96 ( s, HDO); 5.40 (s, CsHsg ^).

Preparation 3

Sodium salt of N-propionylthienamycin To 25 mg of thienamycin in 6 ml of water at 0 ° C., 38.6 mg of sodium bicarbonate, 5 ml of oxane are added in succession and then 1 equivalent of propionic anhydride over a period of 3 minutes with stirring. After 10 minutes the mixture is extracted 3 times with cold ethyl ether. Electrophoresis of the aqueous extract gives (0.05M, pH 7, phosphate buffer, 50 V / cm., 20 min.) And shows that no free thienamycin is present. The aqueous extract is then adjusted to a pH of 6.8 and contains 600 ODU at 302 nm by UV analysis, 95% of which are extinguished after one hour after treatment with hydroxylamine.

NMR (8, D2O) 1.42 (m, CH2CH3, CHsCH); 2.48 (q,

CH2CH3); 2.86-2.90 (m, CH2), 4.30-4.70 (m, CH β-lactam), 4.86 (HDO).

Preparation 4

Sodium salt of N- (methoxyacetyl) thienamycin To 55 mg of thienamycin in 6 ml of water at 0 ° C., 68 mg of sodium bicarbonate, 6 ml of dioxane and 1.1 equivalents of methoxyacetyl chloride are added in succession over a period of V / 2 minutes. The mixture is then stirred for an additional 10 minutes. The mixture is extracted three times in cold ethyl ether. Electrophoresis of the aqueous extract gives (0.05 M, pH 7 phosphate * buffer, 50 V / cm., 20 min.) And shows that no free thienamycin is present. The aqueous extract is adjusted to a pH of 6.8 and contains 105 ODU at 302 nm by UV analysis, 95% of which are extinguished for 1 hour after treatment with hydroxylamine.

NMR (8, D2O) 1.56 (m, CHCHs), 2.84-3.60 (CH2), 3.72 (s, OCH3), 4.29 (s, OCH2), 4.98 (s, HDO).

Preparation 5

5 lithium salt of N- (p-nitrobenzyloxycarbonylthiena-mycin

To 220 mg of thienamycin in 60 ml of water at 0 ° C., 679 mg of sodium bicarbonate, 60 ml of dioxane and then 1.1 equivalents of p-nitrobenzylchloro-10 formate are added in succession over a period of 1 minute. The mixture is then left to react for 10 minutes and finally extracted three times with cold ethyl ether. Electrophoresis (0.05 M, pH 7, phosphate buffer, 50 V / cm., 20 minutes) proves that no free thienamycin ls is present. The aqueous extract is adjusted to a pH of 2.2 by adding a 1 molar phosphoric acid solution and then extracted three times with ethyl acetate. The ethyl acetate extract is dried over magnesium sulfate, filtered and 0.1 normal lithium hydroxide is added to a pH of 8.2. The final pH is then adjusted to 7.0 with 1 molar phosphoric acid and the sample is lyophilized. The yield is 205 mg (54%).

Preparation 6

25 Preparation of N- (p-nitrobenzyloxycarbonyl) -thiena-mycin p-t-butylbenzyl ester

205 mg of the lithium salt of N- (p-nitrobenzyloxycar-bonyl) thienamycin in 2 ml of hexamethylphosphoramide are reacted with 0.1625 ml of p-t-butylbenzyl-30 bromide for 2 hours. The starting product is insoluble in hexamethylphosphoramide, but dissolves after 30 minutes.

The reaction mixture is diluted with ethyl acetate, washed successively with water, aqueous K2HPO4, 35 water, saturated aqueous sodium chloride, dried over magnesium sulfate, filtered, evaporated, subjected to preparative thin layer chromatography on silica gel; elute with 1: 2 CHCb: EtOAc.

40 Yield 160 mg (58%), Rf 0.38, IR (n film) 2.98 NH and OH 5.63, β-lactam; 5.86 wide ester and urethane; NMR (8, CDCh), 1.24 (s, CHCHs, t-butyl), 2.59-3.27 (m, CHz) 3.83-4.47 (m, CH ß-lactam), 5, 15 (SOCH2C6H4NO2), 5.22 (s OCH2C6H4 t-butyl), 7.45 and 8.12 (AB quartet, J = 8 Hz 45 C6H4NO2).

Preparation 7

Preparation of N- (p-nitrobenzyloxycarbonyl) -Thiena-mycin m-phenoxybenzyl ester 50 Following the procedure of preparation 6, the compound mentioned in the title is prepared, but an equivalent amount of m-phenoxybenzyl bromide is used instead of pt-butylbenzyl bromide, as in Example 6.

55 The yield is 11%, IR (u film) 3.0 NH2 and OH 5.63 β-lactam; 5.86 wide peak ester and urethane; nmr (8 CHCb) 1.33 (d CHCHa J = 6); 2.60-3.62 (m, CH2), 7.45 and 8.13 (AB quartet, J = 8, C6H4NO2); 7.26 (s CsmOCôHs) M.S. m / e589,559,547,183.

60

Production of:

Preparation 8

1.) N- (0-Formyl-D-mandeloyl-thienamycin-p-t-butyl-65 benzyl ester

2.) N- (0-formyl) -D-mandeloyl-thienamycin-m-phenoxybenzyl ester

3.) N-D-Mandeloyl-thienamycin-p-t-butylbenzyl ester

633798

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4.) N-D-mandeloyl-thienamycin-m-phenoxybenzyl ester

5.) N-propionyl-thienamycin-p-t-butylbenzyl ester

6.) N-propionyl-thienamycin-m-phenoxybenzyl ester

7.) N-methoxyacetyl-thienamycin-p-t-butylbenzyl ester

8.) N-methoxyacetyl-thienamycin-m-phenoxybenzyl ester

Compounds 1, 3, 5, 7, 2, 4, 6 and 8 can be prepared according to the method of preparations 6 and 7 if the corresponding N-acyl-thienamycin starting material of preparations 1 to 4 is equivalent to the starting material N - (p-Nitrobenzyloxycarbonyl) -thienamycin of preparations 6 and 7 replaced.

Preparation 9

Preparation of N- (p-methoxybenzyloxycarbonyl) -thie-namycin-p-t-butylbenzyl ester

Step A: sodium salt of N- (p-methoxybenzyloxycarbonyl) thienamycin (I) and lithium salt (II)

To 20 mg of thienamycin in 5 ml of water at 0 ° C., 105 mg of sodium carbonate (20 equivalents), 5 ml of dioxane and then 10 equivalents of p-methoxybenzyl chloroformate are added dropwise with stirring for one minute. To

For 15 minutes the pH is adjusted to 7.5 by adding 1 molar phosphoric acid and the solution is extracted three times with ether. In the aqueous part, the pH is adjusted to 2.2 at 0 ° C. and extracted three times with ethyl acetate. The ethyl acetate extract is quickly dried over magnesium sulfate, filtered and extracted with a few ml of water containing 6.3 mg of sodium bicarbonate. The aqueous extract, which was lyophilized, contains 172 ODU at 303 nm by UV analysis in H2O at pH 7.0, 95% being extinguished for one hour after treatment with hydroxylamine. The yield is

16 mg. Electrophoresis (50 V / cm, 20 min., PH 7 aqueous phosphate, 0.05M) shows a spot by bioautography 4 cm in the direction of the anode.

NMR (6, D2O): 1.49 (d, J = 6Hz CHjCH); 2.8-3.7 (m, CH2); 3.99 (s, OMe): 4.0-4.6 (m, β-lactam CH); 4.92 (s, HDO); 5.20 (s, OCH2); 7.13 (d, J = 8Hz CôH »),

The lithium salt II is prepared in the same way, but the ethyl acetate solution is extracted with 0.1 normal lithium hydroxide to a pH of 7.8 (instead of aqueous bicarbonate) and then lyophilized. The spectral and electrophoretic properties of II are the same as compound I.

Step B: N- (p-methoxybenz.yloxycarbonyl) thienamycin-p-t-benzyl ester

37 mg of the lithium salt from stage A in 0.4 ml of hexamethylphosphoramide are reacted with 0.033 ml of p-t-butylbenzyl bromide for 2½ hours. The lithium salt is insoluble in hexamethylphosphoramide, but goes into solution after 15 reaction times.

The reaction mixture is diluted with ethyl acetate, washed successively twice with water, aqueous K2HPO4, water and concentrated saline, dried over magnesium sulfate, filtered, evaporated and subjected to preparative thin layer chromatography on silica gel, eluting with 1: 2 CHCb-EtOAc, giving 47 mg of the receives pure compound II, Rf = 0.3.TR (n, film): 3.0, NH; 5.63, β-lactam; 5.87 wide, ester and urethane. NMR (5, CDCl.i): 1.21 (s, Me and t-butyl); 2.6-3.6 (m, CH2); 3.72 (s, OMe); 3.8-4.4 (m, β-lactam CH);

MS: 582,538,496.

Preparation 10

ls production of N- (p-methoxybenzyloxycarbonyl) thie-namycinbenzylhydryl ester

To 23.5 g of thienamycin in 5 ml of water are added 4 ml of dioxane, 62 mg of sodium bicarbonate, and then in portions at 0 ° C. with stirring, 4 equivalents of 20 p-methoxybenzyl chloroformate over 4 minutes. After a total reaction time of 10 minutes, the pH is adjusted to 7.0 by adding 1 molar phosphoric acid and the mixture is extracted three times with ether. Electrophoresis of the aqueous portion (0.05 M pH 7 aqueous phosphate buffer, 25 50 V / cm, 20 minutes shows 50% conversion to N- (p-methoxybenzyloxycarbonyl) thienamycin.

By adding 1 molar phosphoric acid at 0 ° C., the aqueous solution is adjusted to a pH of 2.2 and extracted three times with ethyl acetate. The ethyl acetate solution is treated with 50 mg diphenyldiazomethane, evaporated and taken up in CH3CN. More diphenyldiazomethane is added to achieve a permanent purple color. After half an hour the solution is evaporated and chromatographed on silica gel and eluted with 1: 2 35 CHCb-EtOAc. 10 mg of the pure title compound Rf 0.25 are obtained. IR (jx, film): 3.0, NH; 5.63, β-lactam; 5.85, 5.89, ester and urethane.

NMR (8, CDCb); 1.23 (s, OH); 1.30 (d, J = 6Hz, CH3CH); 40 2.6-3.6 (m, CH2); 3.78 (s, OMe); 5.02 (s OCH2); 3.8-4.4 (m, β-lactam CH); 6.9 and 7.35 (AB Quartet, J = 9Hz, CsH *), 7.3s CHPhj.

Preparation 11

45 Production of N- (o-nitrobenzyloxycarbonyl) thienamy cinbenzyl ester

Stage A: sodium salt of N- (o-nitrobenzyloxycarbonyl) thienamycin so 43 mg of thienamycin are added to 10 ml of 1: 1-tetrahydrofuran (tetrahydrofuran: water) at 0 ° C. The mixture is then stirred rapidly while adding 102 mg of sodium bicarbonate (10 equivalents) and then 4 equivalents of ss o-nitrobenzyl chloroformate are added dropwise with stirring over 2 minutes. After 30 minutes, the pH is adjusted to 7 by adding aqueous 25% phosphoric acid and the solution is extracted three times with ether. The aqueous layer is evaporated in vacuo at 25 ° C. and then adjusted to a pH of 2.2 at 0 ° C. Solid sodium chloride is added and then the cold acidic solution is extracted three times with cold methyl acetate. The ethyl acetate extracts are combined and quickly washed again with cold saturated saline, dried over magnesium sulfate, filtered and extracted with 65 corn with 10 ml of water which contains 1.75 equivalents of solid sodium bicarbonate. The extract is lyophilized in vacuo at 25 ° C., giving the compound mentioned in the title *.

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Step B: N- (o-nitrobenzyloxycarbonyl) thienamycin benzyl ester

The product from stage A in 7.5 ml of ethyl acetate (from the pH 2.2 extraction) is then treated with an excess of phenyldiazomethane (4 ml of the solution containing 20 mg / ml of ether) at 4 ° C. for 2.3 hours treated long. The mixture is concentrated at 20 ° C. under reduced pressure to give a wet residue. The desired compound is isolated by thin layer chromatography, ethyl acetate: ether (9: 1) and 17.5 mg of N- (o-nitrobenzyloxycarbonyl) - io thienamycin-benzyl ester are obtained.

Preparation 12

Preparation of N- (o-nitrobenzyloxycarbonyl) thiena-

mycin-p-methoxybenzyl ester

To 70 mg of N- (o-nitrobenzyloxycarbonyl) thienamycin in 8 ml of ethyl acetate are added 4 ml of p-methoxyphenyldiazomethane (9 mg / ml acetonitrile) at 4 ° C. The mixture is stirred at 4 ° C for VA hours and then concentrated to a wet paste under reduced pressure at a temperature of 20 ° C. 42 mg of the compound mentioned in the title are obtained by thin layer chromatography on silica gel and by elution with ethyl acetate: ether (9: 1).

Preparation 13

Preparation of N- (o-nitrobenzyloxycarbonyl) -thiena-mycin-p-bromo-phenacyl ester

--0 (TMS) Th4-NH (TMS)

L COO (TMS)

- ^

OH

Th - NHC-O-CH

NO.

C-O-CH II 0

rO

2 "if"

'\ -Br

II

where TMS means the trimethylsilyl radical.

To 24 mg of Th (TMS) 3, compound of formula I, which is prepared by the method of preparation 17, in 0.8 ml of dry tetrahydrofuran are added 23 mg of o-nitrocarbo-benzyloxychloride, followed by 0.015 ml of triethylamine. After the vibration mixing for 30 minutes at 25 ° C, the mixture obtained is concentrated with a dry nitrogen stream to a paste-like residue and then washed three times with petroleum ether. The residue is suspended in 1 ml of dry tetrahydrofuran and 14 mg of p-bromophenacyl bromide are added, followed by the addition of 0.03 ml of triethylamine. After vibration mixing for 30 minutes at 25 ° C, the mixture obtained is evaporated to dryness at 20 ° C in vacuo. The residue is dissolved in 2 ml of ethyl acetate and shaken with 0.3 ml of a pH-4 buffer for 5 minutes. The organic layer is dried over magnesium sulfate, filtered, evaporated to a paste-like residue and then the desired product (44 mg) is isolated by preparative thin layer chromatography on silica gel and eluted with ethyl acetate: chloroform (7: 3).

Preparation 14

Production of N- (TrichloräthoxycarbonyI) thienamy cinbenzylester

Step A: Lithium salt of N- (trichloroethoxycarbonyl) thienamyein

225 mg of sodium carbonate (15.2 equivalents) are added to 40 mg of thienamycin in 18 ml of 1: 1 tetrahydrofuran water at 0 ° C with stirring, and 1.8 equivalents of trichloroethyl chloroformate, dissolved in 0, are then added dropwise with stirring over a period of 2 minutes , 6 ml of tetrahydrofuran. After 6 minutes the pH is adjusted to 7.2 by adding aqueous 25% phosphoric acid and then the solution is extracted with ether. The aqueous part, after removal of ether with traces of ether in vacuo, is then adjusted to a pH of 2.5 at 0 ° C. and extracted with cold ethyl acetate. The ethyl acetate extracts are combined

quickly washed again in cold saline, dried over anhydrous magnesium sulfate, filtered and extracted again with 0.01 normal lithium hydroxide to a pH of 6.8. The aqueous extract is freed of ethyl acetate and also traces of ethyl acetate in vacuo and lyophilized. The residue contains 936 ODu (39.7%) UV analysis at 302 nm, which is extinguished after treatment with hydroxylamine for one hour in a 0.05 molar phosphate buffer (pH 7). The yield is 32 mg. Electrophoresis (50 volts / cm. 20 min., PH 7, 35 aqueous phosphate 0.05M) shows a zone by autobiography (MB 108, staph. Aureus), 2.4 cm in the direction of the anode. Liquid chromatography Cis Bondapak (Waters Assoc.) In aqueous 10% tetrahydrofuran gives a main maximum that is free of unreacted Thiena-40 myein.

Step B: N- (trichloroethoxycarbonyl) thienamycin benzyl ester

32 mg of the compound obtained in stage A in 2 ml of dry-45 kenem distilled dimethylformamide, which contains 7% hexamethylphosphoramide (dry, pH 6.3), are reacted with 0.015 ml of benzyl bromide at 15 ° C. for 2 hours, allowing the reaction mixture to warm up to a temperature of 25 ° C in the course of the reaction. The reaction mixture is then diluted with ethyl acetate, washed successively with cold water, 1% aqueous sodium bicarbonate, water and cold saturated aqueous sodium chloride solution, dried over magnesium sulfate, filtered, evaporated and subjected to preparative thin layer chromatography on silica gel and then with 1% CH3CN eluted in EtOAc to give 10 mg of the title compound, RF = 0.63; IR (a CHCh) 5.63, β-lactam; 5.78 and 5.88 wide ester and urethane. NMR (5 CDCb) 1.35 (d, Me); 2.8-3.7 (m CH2); 3.51 and 4.27 "o (dd, J = 6Hz, β-lactam CH); 4.79 (s, OCHaCCh) 5.42 s (OCH2CóHs); and 7.41 (m, C6H5).

Preparation 15

Preparation of the methyl and benzyl esters of N-bromace-65 tylthienamycin

Step A: N-bromoacetyl-thienamycin

To a chilled solution of 28.8 mg thienamycin and

633 798

14

> ">. ï g natriî'nicarbonate in 10 ml water and 8 ml dioxane indicates ■ ri. with stirring a solution of 0.25 g bromoacetic anhydride in 2 ml dioxane over a period of 20 minutes. The pH value is at 8 , 0. The mixture is then stirred for an additional 5 minutes and then divided into layers by adding 10 ml of ether and finally the pH is adjusted to 7 by adding 8% of the phosphoric acid The aqueous layer is extracted twice again with ether. The aqueous layer is then evaporated to 0.5 ml under reduced pressure, diluted with 2 ml of water and added to 50 ml of XAD-2 resin. The column is eluted with water first 80 ml and then collect the next 100 ml. Change the solvent to 10% tetrahydrofuran and collect an additional 100 ml. The combined eluates are adjusted to pH 7, evaporated to 5 ml under reduced pressure, then freeze dried to obtain the sodium salt of N-bromoacetyl-thienamycin in 60% yield.

UV / .n: ax302 m Li.

Step B: methyl and benzyl esters of N-bromoacetylthie-namycin

The aqueous solution of the sodium salt is divided into layers by adding ethyl acetate at 0 ° C. and adjusted to a pH of 2. The ethyl acetate phase is separated off and the aqueous phase is extracted with ethyl acetate. The combined ethyl acetate solutions are dried over magnesium sulfate and then treated with a solution of diazomethane. The solvents are evaporated off and the rest is chromatographed on silica gel plates. Rf = 0.11 in 2: 1 ethyl acetate-chloroform. The product has a melting point of 118 to 120 ° C. The mass spectrum shows the following properties: M + at m / e 406 and significant fragments at m / e 362,320,183 and 164.

The corresponding benzyl ester is prepared in a similar manner from phenyldiazomethane and has a melting point of 142 to 143 ° C.

IR spectrum: 5.65} i, 5.89 [j. and 6.1 p .. mass spectrum M + m / e 482 also m / e 438.316.259 and 164.

Preparation 16

Preparation of N- (Bromo-t-butoxycarbonyl) thiena-mycin-p-bromophenacyl ester

Step A: Preparation of the N-bromo-t-butoxycarbonyl-O-TMS-thienamycin-TMS-ester

16 mg of Th (TMS) 3 are dissolved in 0.4 ml of dry tetrahydrofuran, to which 20] il (28 mg, 0.13 mmol) of bromo-t-butylchloroformate had been added (boiling point 35 ° C./0.9 mm) and 8 µ (5.67 mg, 0.057 mmol) of triethylamine (distilled off from barium oxide again). The mixture is shaken at a temperature of 25 ° C for 20 minutes. Then the solvent and the excess of starting materials are evaporated off and the crude desired product is obtained. UVA.CH3CO2CH2CH3 320 nm (E9,000).

Step B: Preparation of N- (bromo-t-butoxycarbonyl) thienamycin

3 mg of the product from stage A are dissolved in 0.5 ml of a phosphate buffer with a pH of 7 and 0.1 ml of tetrahydrofuran and then the solution is left to stand at 25 ° C. for 20 minutes. The solution is then placed in a column (5 ml) of Dowes 50x8 (Na + form) and the eluate fractions are registered by U.V. The desired and correct fractions are cleaned and freeze-dried to obtain the desired product.

UVXS2T 304 nm (e = 9,300); Electrophoresis at 50 V / cm for 20 min in a pH 7.0 buffer shows a single bioactive zone that moves 31.5 mm towards the anode.

Step C: Preparation of N- (bromo-t-butoxycarbonyl) thienamycin-p-bromophenacyl ester

The product from Step B (13 mg, 0.022 mmol) is dissolved in 0.4 ml of tetrahydrofuran. To this solution are added p-bromophenacyl bromide (9.6 mg, 0.035 mmol) and 20 ul (14.4 mg, 0.14 mmol) triethylamine. The mixture is shaken at a temperature of 25 ° C for 30 minutes and then evaporated to dryness. 10 ml of ether are added to the rest and then the mixture is reacted with 0.2 ml of a 0.1 molar phosphate buffer with a pH of 7.0.

The organic layer is separated, dried over sodium sulfate, concentrated up to 0.5 ml and then applied to two 20 × 20 cm, 250 μ silica gel GF thin-layer chromatography plates, which are developed with 20% ethyl acetate in chloroform. The desired product is isolated in an amount of 6.7 mg, ie a yield of 42%, and it has one (Rf = 0.65).

Preparation 17

Production of N-benzyloxycarbonyl-thienamycin and N-benzyloxycarbonyl-thienamycin-benzylcarboxylic acid anhydride r ° H

Th —j-J? -0-CH2—

Lcoor fp == -Jf

IK g ü _2

A solution of 16.6 mg thienamycin in 4 ml of a 0.05M phosphate buffer with a pH of 7 and 2 ml of dioxane, which is located in a three-necked flask, which is equipped with a stirrer, a thermometer, a pH electrode and with a Withdrawal opening of an automatic titrator is cooled in a methanol ice bath to a temperature of -8 ° C. The pH is adjusted to 8.2 by adding 0.2 normal sodium hydroxide in 50% aqueous dioxane and then a solution of 0.015 ml of carbobenzyloxychloride in 2 ml of chloroform is added to the solution. The mixture is stirred at a temperature of -6 ° C. and a pH of 8.2 for 10 minutes, then it is divided into layers by adding ether and the pH is adjusted to 7 by adding normal hydrochloric acid . The layers are separated by centrifugation and the aqueous phase is extracted twice with ether. The aqueous phase is separated off by adding ethyl acetate and then acidified to a pH of 2. The ethyl acetate is separated off and then the aqueous layer is extracted again with ethyl acetate. The combined ethyl acetate layers are washed with a saturated sodium chloride solution, dried over magnesium sulfate and then filtered. The filtrate is stirred with water and the pH is adjusted to 7 by adding a dilute sodium bicarbonate solution. The aqueous phase is separated off and freeze-dried, the sodium salt of N-benzyloxycarbonyl-thiena-

s

10th

15

20th

25th

30th

35

40

45

50

55

60

65

15

633798

mycin receives (10 mg, 46%). Due to the UV spectrum,

X max303 mji, E% 147 (E 6.290) a purity of about 80% is confirmed. Electrophoresis at 50 V / cm for 20 minutes at pH 7, followed by bioautography on S. aureus reveals an inhibition zone at +2.5 cm.

The ethereal extracts of the reaction mixture contain the desired product, namely the N-benzyloxy-carbonyl-thienamycin-benzylcarboxylic anhydride.

UVA, max335 m | i.

Preparation 18

Preparation of the benzyl ester of N-benzyloxycarbonyl-thienamycin

The N-benzyloxycarbonyl-thienamycin (Preparation 17) in ethyl acetate is treated in the same manner as described in Preparation 17, except that an equivalent amount of phenyldiazomethane is added to the dry ethyl acetate solution from extraction at one pH is 2, added, and then the solution is left at 4 ° C for 2 hours. Evaporation to dryness gives the crude N-benzyloxycarbonyl-thienamycin benzyl ester, which can be isolated by thin layer chromatography and has a value of Rf = 0.24, 3: 1 ethyl acetate: chloroform. The product obtained is crystallized from ether.

IR 5.63 (lactam carbonyl); Shoulder 5.8 ji (ester); 5.88 ij. (Urethane carbonyl). UV, dioxane, A, max318 mp, E% (e = 10,900) m / e M + 496.

Preparation 19

Preparation of N-carbomethoxy-thienamycin-p-piva-loyloxybenzyl ester

Step A: N-carbomethoxythienamycin

Thienamycin (49 mg, 148 a mol) is dissolved in 14 ml of a 0.05M phosphate buffer with a pH of 7 and cooled in an ice bath. With stirring, the pH is adjusted to 8.2 using an automatic burette. A solution of methyl chloroformate (46, ul, 600 | i, mol) in p-dioxane (580 ul) is added all at once to the homogeneous solution. The pH is then kept at 8.2 using an automatic burette. After 10 minutes the solution is adjusted to a pH of 7 using a dilute phosphoric acid solution and finally washed three times with equal volumes of diethyl ether. The aqueous solution is then concentrated to 4.5 ml and then chromatographed on an XAD-2 resin column. The product is eluted with an aqueous 5% tetrahydrofuran solution (after elution with water) and finally freeze-dried, and 28.9 mg of the product are obtained.

UV (pH 7 phosphate buffer 0.1 N). X max 303 nm (e 6.450) electrophoresis (20 min., 0, lNpH 7 phosphate buffer, 50V / cm mobility 3.5 cm to the cathode.

Th

OH 0! L

NH-S-

ii O

-COOK

10th

R = —O-CH, C = CH.

d \

CH "

Step A: N-benzenesulfonyl-thienamycin 15 thienamycin (52 mg 148 μmol) is dissolved in 25 ml of a 0.1 normal phosphate buffer with a pH of 7 and then stirred in a ice bath with a magnetic stirrer. The pH is adjusted to 8.2 by the addition of 2.5 normal sodium hydroxide using an automatic burette, and then benzenesulfonyl chloride (227 (i.1.226 µmol) in 500 ml p-dioxane is added all at once. The pH is kept at 8.2 for 30 minutes using an automatic burette and then this value is adjusted to 7.0 by adding dilute aqueous phosphoric acid, the reaction solution is concentrated to 15 ml and chromatographed on XAD-2 Resin (50 cc). The column is eluted with water, then with 10% aqueous tetrahydrofuran to elute the product. The 10% aqueous tetrahydrofuran-30 eluate is then concentrated to a third of the volume and freeze-dried using 28 The electro-phoretic mobility of the product (50 V / cm, 20 min., pH 7 0.05N phosphate buffer) is 3.5 cm in the direction of the cathode.

35

Àmax303 (3.650) in pH 7 0.1N phosphate buffer).

Step B: N-benzenesulfonyl-thienamycin-2-methyl-2-40 propen-1-yl ester

According to a known method and with the replacement of equivalent amounts of N-benzenesulfonyl-thienamycin (in the form of its sodium salt) and 2-methyl-2-propen-l-yl chloride for the sodium salt of N-azidoacetyl-thienamycin 45 or pt-butylbenzyl bromide you get the connection mentioned in the title.

50

55

Th-

Preparation 21

rOTMS ■ NHTMS or Th (TMS) COOTMS

Step B: N-carbomethoxy-thienamycin-p-pivaloyloxy-benzyl ester

The procedure of preparation 16, stage C and by substituting equivalent amounts of: N-carbomethoxy-thiena-mycin (in the form of its sodium salt) and p-pivaloyloxy-benzyl chloride bromide gives the N-carbomethoxy-thie-namycin-p-pivaloyloxybenzyl ester.

Preparation 20

Preparation of N-benzenesulfonyl-thienamycin-2-methyl-2-propen-1-yl ester

Preparation of silylated thienamycin fio 80.0 mg of thienamycin suspended in 40 ml of tetrahydrofuran in a nitrogen atmosphere are concentrated to a volume of 10 ml. Then 1.0 ml of hexamethyldi-silazane and 300 ml of trimethylchlorosilane are added. The mixture is reacted for 20 minutes at a temperature of 25 ° C with vigorous stirring. Finally, the suspension obtained is centrifuged to remove the ammonium chloride. The decanted liquid is evaporated to an oil in a stream of nitrogen for further reactions.

633798

Preparation 22

Preparation of N- (p-nitrobenzyloxycarbonyl) -thiena-mycin- (p-nitrobenzyl) ester

-Oh'

Th - nhr2 -coor3

r2 -ich2- ^ y no2 r3 ^ -ck ^ h ^^ - no,

A mixture of 295 mg of the p-nitrobenzyloxy-carbonyl-thienamycin lithium salt from Example 5 and 0.4 g of p-nitro-benzyl bromide in 3 ml of hexamethylphosphoramide is stirred at a temperature of 25 ° C. for 3 hours. This solution is diluted with 50 ml of ethyl acetate and extracted successively with water (three times), with a phosphate buffer which has a pH of 7 and with a saturated sodium chloride solution. The organic phase is dried over magnesium sulfate and evaporated to 5 ml, causing the product to crystallize. The crystals are collected and washed with ethyl acetate to give 160 mg of N-p-nitrobenzyloxycarbonyl-thiena-mycin-p-nitrobenzyl ester, which has a melting point of 168 to 170 ° C.

NMR (CHCLs) 1.35 [(d), 3 = 6 HZ], 4.0-4.3 and 2.8-3.5 (m), 5.16 [(s), (carbamate OCH2)] ; 5.24 (AB quartet, J = 14) 7.42 (d), 7.56 (d), 8.16 (d) (J = 9, aromatic) IR 5.64 / u (lactam C = 0) .

Preparation 22a

Preparation of N-bromo-t-butyloxycarbonyl-thiena-mycin

Method A: 190 mg of thienamycin are kept at a temperature of 0 ° C. in 15 ml of a 0.1 molar phosphate buffer with a pH of 7.0 and 15 ml of dioxane. The pH of the solution is adjusted to 8.5 to 9.0 by adding 1-normal sodium hydroxide while adding 480 mg of bromo-t-butyl chloroformate over a period of 5 minutes. The mixture is stirred for 30 minutes, then neutralized by adding 1 normal hydrochloric acid to a pH of 7.0 and finally extracted with ether. The aqueous layer is separated off, concentrated to a volume of 10 ml and chromatographed on a Dowex 50x8 (Na form) column (1.5 "x 10") which is eluted with water, 113 mg of the desired product. The solid product is obtained by lyophilizing the solution.

Method B: 95 mg of thienamycin are kept in 10 ml of a 0.1 molar phosphate buffer and 10 ml of dioxane at a temperature of 0 ° C. The solution is adjusted to a pH of 8.5 to 9.0 while adding 240 mg of bromo-t-butylchloroformate. The mixture is then stirred for 30 minutes and acidified to pH 2.0 by the addition of phosphoric acid. The acidified solution is extracted twice with 25 ml of ethyl acetate. The organic layer is separated off and extracted again.

16

corn with 10 ml sodium bicarbonate solution containing 30 mg sodium bicarbonate. The aqueous layer contains 30 mg of the desired product and is lyophilized to give the solid product.

s

NMR (60 MHz, D20) Al, 26 (d), 1.60 (s), 2.65-3.50 (m), 3.70 (s) and 3.90-4.20 (m). UVA. °! ° 303nm.

Preparation of N-bromo-t-butyloxycarbonyl-thiena-10 mycin-p-nitrobenzyl ester

100 mg of the lyophilized sodium salt of N-bromo-t-butyloxycarbonyl-thienamycin are stirred together with 300 mg of p-nitrobenzyl bromide in 2 ml of hexamethylphosphoramide at 25 ° C. for one hour. The mixture is diluted with 10 ml of ethyl acetate and then washed well with water. The organic layer is separated, dried over sodium sulfate and chromatographed on two 250 u silica gel GF thin layer chromatography plates using ethyl acetate as solvent (Rf 20 0.45) to give 50 mg of the desired product. IR (CDCh): 1777 (β-lactam) and 1711 cm- '(ester):

UVA. ™ "270 nm and 322 nm; NMR (CDCh, 60 MHz): ole, 38 (d), 1.58 (s), 2.60-3.80 (m), 3.78 (s), 3.90-4.20 (m), 5.30 (s), 25 7.55 (d) and 8.30 ppm (d).

example 1

Production of Q-methyl-thienamycin

Th-

35

■ OCH-3

-nh2

-COOH

Stage A: 0-methyl-N- (p-nitrobenzyloxycarbonyl) -thie-namycin- (p-nitrobenzyl) ester To a solution of 135 mg of Np-nitrobenzyloxycar-4o bonyl-thienamycin-p-nitrobenzyl ester in 50 ml of methylene chloride at 0 ° C, 0.5 ml of 0.006 molar borofluoric acid in ether methylene chloride (3: 1) is added with vigorous stirring, and 10 ml of a cooled solution of 0.6 molar diazomethane in methylene chloride are then immediately added. The diazomethane is removed within one minute. The solution is extracted with 10 ml of a 0.1 normal phosphate buffer with a pH of 7, dried and evaporated to a small volume. The solution is applied to two 8 "x8" 1000 ml silica gel plates, which are developed with 3: 1 50 ethyl acetate-chloroform. The band at 3 to 4.5 cm gives 12 mg of the recovered starting material. The band has 6 to 8 cm gives 20 mg of crystalline 0-methyl-N- (p-nitrobenzyloxycarbonyl) thienamycin (p-nitrobenzyl) ester, which has a melting point of 172 to ss 174 ° C.

MS m / e 600 (M +), 568,542,500,447,389,304 and 59.

Step B: O-methyl-thienamycin 60 A solution of 20 mg of O-methyl-Np-nitrobenzyloxycar-bonyl-thienamycin-p-nitrobenzyl ester in 2 ml of tetrahydrofuran and 1 ml of ethanol is made at a pressure of about 3.5 kg / cm 2 and at a temperature of 23 ° C in the presence of 20 mg of platinum oxide for 214 hours. The catalyst is filtered off and 1 ml of a 0.1 normal phosphate buffer with a pH of 7 is added to the filtrate. The solution is then evaporated to 2 ml under reduced pressure and the mixture is taken in 5 ml of water and

17th

633 798

5 ml of ethyl acetate and centrifuged. The ethyl acetate layer is removed and then the aqueous layer is extracted again with ethyl acetate and with ether and then filtered through cellite. The aqueous solution is then applied to a column (20 ml) of an XAD-2 resin. The column is eluted first with water and then with 10% tetrahydrofuran. The tetrahydrofuran eluate is then concentrated and lyophilized and essentially pure O-methyl-thienamycin is obtained.

Example 2

Preparation of 0- (methyoxymethyl) thienamycin

T-OCH "OCH, z -1

Th-

NH,

-COOH

Step A: A solution of 58 mg of p-nitrobenzyloxycar-bonyl-thienamycin-p-nitrobenzyl ester in 5 ml of tetrahydrofuran and 1 ml of hexamethylphosphoramide is cooled to -78 ° C. A 2-molar solution of phenyllithium (0.1 ml) and then immediately 0.2 ml of methylchloromethyl ether are added to this solution with stirring. The mixture warms to a temperature of 25 ° C. over an hour. 25 ml of methylene chloride are added and the solution is extracted with 25 ml of a 0.1 normal phosphate buffer which has a pH of 7 and then four times with 25 ml of water each. The methylene chloride solution is evaporated and the rest is triturated with hexane. The residue, which is insoluble in hexane, is chromatographed on silica gel and 0-methoxymethyl-N- (p-nitrobenzyloxycar bonyl-thienamycin (p-nitrobenzyl) ester is obtained.

Step B: After the hydrogenation process of preparation 4, step B, the compound mentioned in the title is obtained if the substitution indicated is carried out.

Example 3

Production of O-methyl-thienamycin "OCH-

Tjl— | ~ NH 2

COOH

Step A: O-methyl-N-benzyloxycarbonyl-thienamycin-benzyl ester

A solution of 5 mg of N-benzyloxycarbonyl-thiena-mycin-benzyl ester (preparation 18) in 0.3 ml of methylene chloride is cooled to 0 ° C. and then 0.1 ml of a 0.006 molar solution of borofluoric acid in 5: 1 ether is added. Methylene chloride was added, followed by the immediate addition of 0.5 ml of 0.1 molar diazomethane in methylene chloride. The solution discolors within a minute. The mixture is stirred with ether and a phosphate buffer which has a pH of 7 and the ethereal phase is evaporated. The residue is chromatographed twice on 8 "250 u silica gel plates in 35% ethyl acetate-chloro-10 form. The band at Rf 0.5 is eluted with ethyl acetate. The measurement of the UV absorption at 318 ml shows a recovery of the original optical density of 13% The mass spectral analysis shows the following results on m / e M + 510 and fragments at m / e 410,59,333 and 478, which are characteristic of O-methyl-N-benzyloxycarbonyl-thienamycin-benzyl-ester .

Alternative stage A: O-methyl-N-benzyloxycarbonyl-thienamycin-benzyl ester 20 A solution of 5 mg of N-benzyloxycarbonyl-thienamycin-benzene ester in 0.2 ml of dry tetrahydrofuran is cooled to -78 ° C. To this solution is added 5 µl of a phenyl lithium solution (2-normal), followed by the immediate addition of 1 µl 1 methyl fluorosulfonate. The mixture is stirred at -78 ° C for 25 minutes and then diluted with 2 ml of ether and extracted with 2 ml of a 0.01 normal phosphate buffer which has a pH of 7. The ethereal layer is evaporated and the residue is chromatographed on a 2 "x8" thin layer chromatography-30 silica gel plate in 1: 1 ethyl acetate-chloroform. The band at Rf = 0.63 is isolated and gives the desired product, namely O-methyl-N-benzyloxycarbonyl-thienamycin-ben-zylester.

35 Step B: O-methylthienamycin

A solution of 0.2 mg of the above-mentioned product in 0.5 ml of dioxane, which contains 0.3% acetic acid, is for 3 hours in the presence of 2 mg of a palladium oxide catalyst at a pressure of about 3.2 kg / cm2 and 40 a temperature of 25 ° C hydrogenated. The mixture is shaken with 4 ml of ether and 1.4 ml of water. The UV absorption of the ether layer (A.max318) shows 50% recovery and the UV absorption of the aqueous layer (Lnax285) 10% of the initial absorption. Electrophoresis of a sample 45 of 5 and 1 of the aqueous layer at a pH of 7.50 V / cm for 20 minutes followed by bioautography of plates inoculated with S. aureus shows two inhibitions: +0.5 cm (19 mm. diameter) and +4 cm (25 mmol). The slower moving spot is O-methyl-N-carbo-50 benzyloxy-thienamycin. Substantially pure O-methyl-thienamycin is obtained from the aqueous phase by chromatography or with X AD-2 resin, using 10% aqueous tetrahydrofuran as the eluting solvent.

B

Claims (3)

633798 2 PATENT CLAIMS 1. Process for the preparation of new compounds of the formula SCH2CH, NH2 COOH (II) and their non-toxic, pharmaceutically acceptable salts, in which R4 is an ether radical, characterized in that compounds of the formula OH X / \ yS = H2Ca2® 3- n ï -i KL 10 15 ✓ "CCOK ^ 25 in which R1 and R2 represent easily removable protective groups, correspondingly etherified, the protective groups removed, and that any compounds obtained are converted into the corresponding salts. 30 2. The method according to claim 1, characterized in that R4 is an optionally substituted alkyl radical, aryl, aralkyl, alkenyl, alkynyl, cycloalkyl, heteroaryl or heteroaralkyl radical 3. The method according to claim 1, characterized in 33 that R4 is a radical of the formula - ( CH2) nZR ", where n is an integer from 1 to 4, R" is hydrogen, amino, mercapto, hydroxy, alkylamino, dialkylamino, alkyl, alkylthio, arylthio, alkoxy, aryloxy, alkenyl, alkynyl, aryl, aralk yl, acylcycloalkyl, heteroaryl or heteroaralkyl and Z is oxygen, sulfur, nitrogen or carbonyl. 4. The method according to claim 1, characterized in that R4 is a radical of the formula -CHR "'R" ", wherein R'" is hydrogen, amino, mercapto, hydroxy, alkylamino, dialkylamino, alkyl, alkylthio, arylthio, alkoxy, aryloxy , Alkenyl, alkynyl, aryl, aralkyl, cycloalkyl, heteroaryl or heteroaralkyl and R "" denotes amino, hydroxy, azido, carbamoyl, guanidino, acyloxy, halogen, sulfamino, tetrazolyl, sulfo, carboxy, carbalkoxy, phosphono, alkoxy or alkylthio is. 5. The method according to claim 1, characterized in that R4 is a radical of the formula - (CH2) mA- (CH2) nY, wherein m and n are zero or 1 to 5, A is O, NR '(R' is hydrogen or lower alkyl with 1-6 C atoms) or S or A represents a simple bond and Y is selected from one of the following groups: 1-imidazolylmethyl, 2-benzofuranylmethyl, 2-indolyl-methyl, 2-phenylvinyl, 2-phenylethynyl, 1-aminocyclohexyl, 1.) Amino or substituted amino: © -N (R> 2 and -N (R> 3 wherein the values for R are independently selected from the following group: hydrogen, N (R ') 2 (R' is hydrogen or lower alkyl having 1-6 C atoms), lower alkyl or lower alkoxy each having 1 to 6 carbon atoms, lower alkoxy lower -alkyl in which the alkoxy part has 1 to 6 carbon atoms and the alkyl part contains 2 to 6 carbon atoms; Cycloalkyl and cycloalkylalkyl, wherein the cycloalkyl part has 3-6 carbon atoms and the alkyl part contains 1 to 3 carbon atoms; 2 groups R together with the nitrogen atom to which they are attached can form a ring with 3 to 6 atoms; 2.) Amidino and substituted amidino: -N = C-N (R) 2 R wherein R is independently selected from the following group: hydrogen; N (R ') 2 (R' is hydrogen or lower alkyl with 1-6 C atoms); Lower alkyl or lower alkoxy, each with 1-6 carbon atoms, lower alkoxy lower alkyl, the alkoxy part containing 1-6 carbon atoms and the alkyl part containing 2-6 carbon atoms (if the lower alkoxy lower alkyl radical is bonded to the carbon atom, the alkyl part contains 1 -6 carbon atoms); Cycloalkyl and cycloalkylalkyl, in which the alkyl part contains 1-3 carbon atoms, two groups R together with the atoms to which they are attached can form a ring with 3-6 atoms; 3.) Guanidino and substituted guanidino: -NH-C-N (R> II NO 40 wherein R is as defined above under 2.); 4.) Guanyl and substituted guanyl: 45 -C = NO I. N (R> wherein R is defined above under 2.); so 5.) Nitrogen-containing mono- and bicyclic heterocycles (aromatic and non-aromatic) with 4-10 core atoms, in which the heteroatom or the heteroatoms can also be oxygen and sulfur in addition to nitrogen. 55 6. The method according to claim 2, characterized in that R4 is one of the following radicals: Benzyl, p-hydroxybenzyl, 4-amino-4-carboxybutyI, 60 methyl, cyanomethyl, 2-pentenyl, n-amyl, n-heptyl, ethyl, 3- and 4-nitrobenzyl, phenethyl, ß, ß-diphenylethyl, methyl-diphenylmethyl, triphenylmethyl, 2-methoxyphenyl, 2,6-dimethoxyphenyl, 2,4,6-trimethoxyphenyl, 3,5-dimethyl-4-isoxazolyl, 3 Butyl-5-methyl-4-isoxazolyl, 65 5-methyl-3-phenyl-4-isoxazolyl, 3- (2-chlorophenyl) -5-methyl-4-isoxazolyl, 3- (2,6-dichlorophenyl) -5-methyl-4-isoxazolyl, D -4-amino-4-carboxybutyl, D-4-N-benzoylamino- 4-carboxy-n-butyl, p-aminobenzyl, o-aminobenzyl, m-ami- 633798 nobenzyl, p-dimethylaminobenzyl, (3-pyridyl) methyl, 2-ethoxy-l-naphthyl, 3-carboxy-2-quinoxalinyl, 3- (2,6-di-chlorophenyl) -5- (2-furyl) -4 -isoxazolyl, 3-phenyl-4-isoxazolyl, 5-methyl-3- (4-guanidinophenyl) -4-isoxazolyl, 4-guanidinomethylphenyl, 4-guanidinomethylbenzyl, 4-guanidi-nobenzyl, 4-guanidinophenyl, 2 , 6-dimethoxy-4-guanidino-phenyl, o-sulfobenzyl, p-carboxymethylbenzyl, p-carbamoylmethylbenzyl, m-fluorobenzyl, m-bromobenzyl, p-chlorobenzyl, p-methoxybenzyl, 1-naphthylmethyl, 3-isothiazolyl -methyl, 4-isothiazolylmethyl, 5-isothiazolylmethyl, guanyl-thiomethyl, 4-pyridylmethyl, 5-isoxazolylmethyl, 4-methoxy-5-isoxazolylmethyl, 4-methyl-5-isoxazolylmethyl, 2- and 3-thienylaminomethyl, 2- (5-nitrofuranyl) vinyl, phenyl, o-methoxyphenyl, o-chlorophenyl, o-phenylphenyl, p-aminomethylbenzyl, l- (5-cyanotriazolyl) methyl, difluoromethyl, dichloromethyl, dibromomethyl, 1- (3-methylimidazolyl) methyl, 2- or 3- (5-carboxymethylthienyl) methyl, 2- or 3- (4-carbamoylthienyl) methyl, 2- or 3- (5-methylthienyl) methyl, 2- or 3- (5-methoxythienyl) methyl, 2- or 3- (4-chlorothienyl) methyl, 2- or 3- (5-sulfothienyl) methyl, 2- or 3- (5- Carboxythienyl) methyl, 3- (l, 2,5-thia-diazolyl) methyl, 3- (4-methoxy-1,2,5-thiadiazolyl) methyl, 2-furylmethyl, 2- (5-nitrofuryl) -methyl, 3-furylmethyl, 2-thienylmethyl, 3-thienylmethyl, tetrazolylmethyl, benzamidinomethyl and cyclohexylamidinomethyl. oh l A / \ / CH2C; '2Ä ;?' 'Ï w - j