N- And carboxyl derivatives of thienamycin

Abstract

Disclosed are N-acyl and carboxyl derivatives of the antibiotic thienamycin, which has the following structure: ##STR1## Such derivatives are useful as antibiotics. Also disclosed are processes for the preparation of such derivatives, pharmaceutical compositions comprising such derivatives and methods of treatment comprising administering such derivatives and compositions when an antibiotic effect is indicated.

Description

BACKGROUND OF THE INVENTION

This invention relates to certain N-acyl and carboxyl derivatives of the new antibiotic thienamycin. Such derivatives are useful as antibiotics. This invention also relates to processes for the preparation of such compounds, pharmaceutical compositions comprising such compounds, and methods of treatment comprising administering such compounds and compositions when an antibiotic effect is indicated.

Thienamycin (I), is disclosed and claimed in co-pending, commonly assigned U.S. patent application Ser. No. 526,992, filed Nov. 25, 1974 (now U.S. Pat. No. 3,950,357, issued Apr. 13, 1976), which patent is incorporated herein by reference since thienamycin may serve as a starting material for the compounds of the present invention. ##STR2##

Thienamycin and all of its isomers (in pure form and as mixtures) are also obtainable by the total synthesis disclosed and claimed in co-pending, commonly assigned U.S. patent application Ser. No. 833,210 (Sept. 15, 1977). This application is incorporated herein by reference to the extent that it makes available all isomers of I as starting materials in the preparation of the compounds of the present invention.

The N-acylated, carboxyl derivatives of thienamycin of the present invention may be depicted by the following generic structural formula (II): ##STR3## or, more conveniently, by the symbol: ##STR4## wherein "Th" symbolizes the bicyclic nucleus of Thienamycin and the OH, amino and carboxyl groups of thienamycin are illustrated.

R.sup.1 and R.sup.2 are independently selected from the group consisting of hydrogen (R.sup.1 and R.sup.2 are not both hydrogen) or an acyl group. The most preferred compounds of this invention are those wherein R.sup.1 is hydrogen and R.sup.2 is acyl. The term "acyl" is by definition inclusive of the alkanoyls including derivatives and analogues thereof such as thio analogues wherein the carbonyl oxygen is replaced by sulphur, diacyl radicals wherein R.sup.1 and R.sup.2 are joined together; as well as the sulphur and phosphorous acyl analogues such as substituted sulfonyl-, sulfinyl- and sulfenyl-radicals, and substituted P (III and V) radicals such as the substituted phosphorous-, phosphoric-, phosphonous- and phosphonic-radicals, respectively. Such acyl radicals of the present invention are further defined below.

X is oxygen, sulphur or NR' (R'.dbd.H or R); and R is, inter alia, representatively selected from the group consisting of conventional blocking groups such as trialkylsilyl, substituted and unsubstituted benzyl, acyl and the pharmaceutically acceptable salt, ester and amide moieties known in the bicylic .beta.-lactam antibiotic art; the definition of R is given in greater detail below.

There is a continuing need for new antibiotics for unfortuantely there is no static effectiveness of a given antibiotic because continued wide scale usage of any such antibiotic selectively gives rise to resistant strains of pathogens. In addition, the known antibiotics suffer from the disadvantage of being effective only against certain types of microorganisms. Accordingly, the search for new antibiotics continues.

Unexpectedly, it has been found that the compounds of the present invention are broad spectrum antibiotics, which are useful in animal and human therapy and in inanimate systems.

Thus, it is an object of the present invention to provide a novel class of antibiotics which possess the basic nuclear structure of the antibiotic thienamycin but which are characterized as N-acylated, carboxyl derivatives thereof. These antibiotics are active against a broad range of pathogens which representatively include both gram positive bacteria such as S. Aureus, B. subtilis, and Strept. pyogenes and gram negative bacteria such as E. coli, and Salmonella. Further objects of this invention are to provide chemical processes for the preparation of such antibiotics and their non-toxic pharmaceutically acceptable salts; pharmaceutical compositions comprising such antibiotics; and to provide methods of treatment comprising administering such antibiotics and compositions when an antibiotic effect is indicated.

DETAILED DESCRIPTION OF THE INVENTION

The compounds of the present invention (II) are conveniently prepared in the following sequence: first thienamycin (I) is N-acylated to form the N-acyl intermediate (1) which is then esterified to form the desired product (II): ##STR5## wherein all symbolism is as previously defined. The above preparation is described in greater detail below. However, it is to be recognized that the two steps may be conducted in reverse order, or may be effected simultaneously.

In the generic representation of the compounds of the present invention (II, above), the acyl radical represented by either R.sup.1 or R.sup.2 can, inter alia, be substituted or unsubstituted aliphatic, aromatic or heterocyclic, araliphatic or heterocylylaliphatic carboxylic acid radical, a substituted or unsubstituted carbamyl radical or a carbothioic acid radical. One group of acyl radicals can be represented by the general formula: ##STR6## wherein X is O or S and R" represents hydrogen; amino, substituted amino such as alkyl- and dialkylamino wherein the alkyl radical comprises 1 to about 6 carbon atoms; substituted or unsubstituted: straight or branched chain alkyl wherein the alkyl radical comprises 1 to about 6 carbon atoms; mercapto such as alkylthio, typically comprising 1 to 6 carbon atoms; arylthio, typically comprising 6 to 10 carbon atoms; hydroxy such as alkoxy, typically comprising 1 to 6 carbon atoms; aryloxy, typically comprising 6 to 10 carbon atoms; alkenyl, or alkynyl groups typically comprising 2 to 6 carbon atoms; aryl such as phenyl; aralkyl such as benzyl; cycloalkyl, typically comprising 3 to 6 carbon atoms; or a heteroaryl or heteroaralkyl group (mono- and bicyclic) wherein the alkyl moiety typically comprises 1 to 3 carbon atoms and the heterocyclic ring comprises typically 4-10 atoms and the hetero atom or atoms are selected from O,N and S; such above-listed groups can be unsubstituted or can be substituted by radicals such as OH, SH, SR (R is loweralkyl or aryl such as phenyl), alkyl or alkoxy groups having 1 to about 6 carbon atoms, halo, such as Cl, Br, F and I, cyano, carboxy, sulfamino, carbamoyl, sulfonyl, azido, amino, substituted amino such as alkylamino including quaternary ammonium wherein the alkyl group comprises 1 to 6 carbon atoms, haloalkyl such as trifluoromethyl, carboxyalkyl, carbamoylalkyl, N-substituted carbamoylalkyl, wherein the alkyl moiety of the foregoing four radicals comprises 1 to about 6 carbon atoms, amidino, guanidino, N-substituted guanidino, guanidino lower alkyl and the like. Representative examples of such acyl groups that might be mentioned are those wherein R" is benzyl, p-hydroxybenzyl, 4-amino-4-carboxybutyl, methyl, cyanomethyl, 2-pentenyl, n-amyl, n-heptyl, ethyl, 3- or 4-nitrobenzyl, phenethyl, .beta.,.beta.-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-carboxybutyl, D-4-N-benzoylamino-4-carboxy-n-butyl, p-aminobenzyl, o-aminobenzyl, m-aminobenzyl, p-dimethylaminobenzyl, (3-pyridyl)methyl, 2-ethoxy-1-napthyl, 3-carboxy-2-quinoxalinyl, 3-(2,6-dichlorophenyl)-5-(2-furyl)-4-isoxazolyl, 3-phenyl-4-isoxazolyl, 5-methyl-3-(4-guanidinophenyl)-4-isoxazolyl, 4-guanidinomethylphenyl, 4-guanidinomethylbenzyl, 4-guanidinobenzyl, 4-guanidinophenyl, 2,6-dimethoxy-4-guanidino, o-sulfobenzyl, p-carboxymethylbenzyl, p-carbamoylmethylbenzyl, m-fluorobenzyl, m-bromobenzyl, p-chlorobenzyl, p-methoxybenzyl, 1-naphthylmethyl, 3-isothiazolylmethyl, 4-isothiazolylmethyl, 5-isothiazolylmethyl, guanylthiomethyl, 4-pyridylmethyl, 5-isoxazolylmethyl, 4-methoxy-5-isoxazolylmethyl, 4-methyl-5-isoxazolylmethyl, 1-imidazolylmethyl, 2-benzofuranylmethyl, 2-indolylmethyl, 2-phenylvinyl, 2-phenylethynyl, 1-aminocyclohexyl, 2- and 3-thienylaminomethyl, 2-(5-nitrofuranyl)vinyl, phenyl, o-methoxyphenyl, o-chlorophenyl, o-phenylphenyl, p-aminomethylbenzyl, 1-(5-cyanotrizolyl)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-(methoxythienyl)methyl, 2- or 3-(4-chlorothienyl)-methyl, 2- or 3-(5-sulfothienyl)methyl, 2- or 3-(5-carboxythienyl)methyl, 3-(1,2,5-thiadiazolyl)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.

The acyl group can also be a radical of the formula: ##STR7## wherein X is O or S and n is 0-4, Z represents oxygen, sulfur, carbonyl or nitrogen and R" is defined as above. Representative members of the substituent

--(CH.sub.2).sub.n ZR" that might be mentioned are allylthiomethyl, phenylthiomethyl, butylmercaptomethyl, .alpha.-chlorocrotylmercaptomethyl, phenoxymethyl, phenoxyethyl, phenoxybutyl, phenoxybenzyl, diphenoxymethyl, dimethylmethoxymethyl, dimethylbutoxymethyl, dimethylphenoxymethyl, 4-guanidinophenoxymethyl, 4-pyridylthiomethyl, p-(carboxymethyl)phenoxymethyl, p-(carboxymethyl)phenylthiomethyl, 2-thiazolylthiomethyl, p-(sulfo)phenoxymethyl, p-(carboxymethyl)phenylthiomethyl, 2-pyrimidinylthiomethyl, phenethylthiomethyl, 1-(5,6,7,8-tetrahydronaphthyl)oxomethyl, N-methyl-4-pyridylthio, benzyloxy, methoxy, ethoxy, phenoxy, phenylthio, amino, methylamino, dimethylamino, pyridinium methyl, trimethylammonium-methyl, cyanomethylthiomethyl, trifluoromethylthiomethyl, 4-pyridylethyl, 4-pyridylpropyl, 4-pyridylbutyl, 3-imidazolylethyl, 3-imidazolylpropyl, 3-imidazolylbutyl, 1-pyrroloethyl, 1-pyrrolopropyl and 1-pyrrolobutyl.

Alternatively, the acyl group can be a radical of the formula: ##STR8## wherein R" is defined as above and R"' is a radical such as amino, hydroxy, azido, carbamoyl, guanidino, amidino, acyloxy, halo, such as Cl, F, Br, I, sulfamino, tetrazolyl, sulfo, carboxy, carbalkoxy, phosphono and the like. Representative members of the substituent ##STR9## that might be mentioned are .alpha.-aminobenzyl, .alpha.-amino-(2-thienyl)methyl, .alpha.-(methylamino)benzyl, .alpha.-amino-methylmercaptopropyl, .alpha.-amino-3- or 4-chlorobenzyl, .alpha.-amino-3- or 4-hydroxybenzyl, .alpha.-amino-2,4-dichlorobenzyl, .alpha.-amino-3,4-dichlorobenzyl, D(-)-.alpha.-hydroxybenzyl, .alpha.-carboxybenzyl, .alpha.-amino-(3-thienyl)methyl D-(-)-.alpha.-amino-3-chloro-4-hydroxybenzyl, .alpha.-amino(cyclohexyl)methyl, .alpha.-(5-tetrazolyl)-benzyl, 2-thienyl-carboxymethyl, 3-thienyl-carboxymethyl, 2-furylcarboxymethyl, 3-furyl-carboxymethyl, .alpha.-sulfaminobenzyl, 3-thienyl-sulfaminomethyl, .alpha.-(N-methylsulfamino)-benzyl, D(-)-2-thienyl-guanidinomethyl, D(-)-.alpha.-guanidinobenzyl, .alpha.-guanylureidobenzyl, .alpha.-hydroxybenzyl, .alpha.-azidobenzyl, .alpha.-fluorobenzyl, 4-(5-methoxy-1,3-oxadiazolyl)-aminomethyl, 4-(5-methoxy-1,3-oxadiazolyl)-hydroxymethyl, 4-(5-methoxy-1,3-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)-aminomethyl, 2-(1,4-thiazolyl)-hydroxymethyl, 2-(1,4-thiazolyl)-carboxymethyl, 2-benzothienylaminomethyl, 2-benzothienylhydroxymethyl, 2-benzothienylcarboxymethyl, .alpha.-sulfobenzyl, .alpha.-phosphonobenzyl, .alpha.-diethylphosphono and .alpha.-monoethylphosphono. Further acyl radicals of interest in this class when X=oxygen are: ##STR10## wherein R.sup.3 and R.sup.4 are as defined below. R.sup.3 represents hydrogen, halo, such as chloro, fluoro, bromo, iodo, amino, guanidino, phosphono, hydroxy, tetrazolyl, carboxy, sulfo, or sulfamino and R.sup.4 represents phenyl, substituted phenyl, a mono- or bicyclic heterocyclyl containing one or more oxygen, sulfur or nitrogen atoms in the ring, such as furyl, quinoxalyl, thienyl, quinolyl, quinazolyl, thiazolyl, isothiazolyl, tetrazolyl, oxadiazolyl, thiadiazolyl and the like substituted heterocycles, phenylthio, phenyloxy, lower alkyl of 1-6 carbon atoms, heterocyclic or substituted heterocyclic thio groups; or cyano. The substituents on the moieties, R.sup.3 and R.sup.4, can be halo, carboxymethyl, guanidino, guanidinomethyl, carboxamidomethyl, aminoethyl, nitro, methoxy or methyl. When R.sup.3 is selected from the group consisting of hydrogen, hydroxy, amino or carboxy and R.sup.4 is selected from the group consisting of phenyl, or a 5- or 6-membered heterocyclic ring having one or two sulfur, oxygen or nitrogen hetero atom such as tetrazolyl, thienyl, furyl and phenyl, the following acyl radicals are representative: phenylacetyl, 3-bromophenylacetyl, p-aminomethylphenylacetyl, 4-carboxymethylphenylacetyl, 4-carboxyamidomethylphenylacetyl, 2-furylacetyl, 5-nitro-2-furylacetyl, 3-furylacetyl, 2-thienylacetyl, 5-chloro-2-thienylacetyl, 5-methoxy-2-thienylacetyl, .alpha.-guanidino-2-thienylacetyl, 3-thienylacetyl, 2-(4-methylthienyl)acetyl, 3-isothiazolylacetyl, 4-methoxy-3-isothiazolylacetyl, 4-isothiazolylacetyl, 3-methyl-4-isothiazolylacetyl, 5-isothiazolylacetyl, 3-chloro-5-isothiazolylacetyl, 3-methyl-1,2,5-oxadiazolylacetyl, 1,2,5-thiadiazolyl-4-acetyl, 3-methyl-1,2,5-thiadiazolylacetyl, 3-chloro-1,2,5-thiadiazolylacetyl, 3-methoxy-1,2,5-thiadiazolylacetyl, phenylthioacetyl, 4-pyridylthioacetyl, cyanoacetyl, 1-tetrazolylacetyl, .alpha.-fluorophenylacetyl, D-phenylglycyl, 4-hydroxy-D-phenylglycyl, 2-thienylglycyl, 3-thienylglycyl, phenylmalonyl, 3-chlorophenylmalonyl, 2-thienylmalonyl, 3-thienylmalonyl, .alpha.-phosphonophenylacetyl, .alpha.-amino cyclohexadienylacetyl, .alpha.-sulfaminophenylacetyl, .alpha.-hydroxyphenylacetyl, .alpha.-tetrazolylphenylacetyl and .alpha.-sulfophenylacetyl.

The acyl substituent R.sup.1 and R.sup.2 may also be selected from sulphur (1) and phosphorous (2) radicals: ##STR11## wherein with respect to (1), m and n are integers selected from 0 or 1 and Y=O.sup..crclbar. M.sup..sym., --N(R").sub.2, and R"; wherein M.sup..sym. is selected from hydrogen alkali metal cations and organic bases; and R" is as defined above, e.g., alkyl, alkenyl, aryl and heteroaryl. With respect to (2) X=O or S; n=0 or 1; and Y' and Y" are selected from the group consisting of O.sup..crclbar. M.sup..sym., --N(R").sub.2, R" and ZR" wherein all symbolism is as defined above, e.g., R" and ZR" are representatively: alkyl, alkenyl, aryl, heteroaryloxy; Y' and Y", including R" moieties, can be joined together to form cyclic ester, ester-amide and amide functions. Illustrative examples of (1) are N-(methylsulphonyl)thienamycin, N-(o-nitrophenylsulphonyl)thienamycin, N-(p-chlorophenylsulphinyl)thienamycin, N-(o-nitrophenylsulphenyl)thienamycin, N-sulphamoylthienamycin, N-dimethylsulphamoylthienamycin and thienamycin N-sulphonic acid sodium salt. Illustrative examples of (2) are N-(dimethoxyphosphino)thienamycin, N-(dibenzyloxyphosphino)thienamycin, N-(dihydroxyphosphino)thienamycin disodium salt, N-dimethoxyphosphinyl)thienamycin, N-(dimethoxyphosphinothioyl)thienamycin, N-(dibenzyloxyphosphinyl)thienamycin, N-(dihydroxyphosphinyl)thienamycin disodium salt.

An acyl class of particular interest is those acyl radicals, R.sup.1 and R.sup.2 of Structure II, above, which are selected from the group consisting of conventionally known N-acyl blocking or protective groups such as carbobenzyloxy, ring-substituted carbobenzyloxy such as o- and p-nitrocarbobenzyloxy, p-methoxycarbobenzyloxy, chloroacetyl, bromoacetyl, phenylacetyl, t-butoxycarbonyl, trifluoroacetyl, bromoethoxycarbonyl, 9-fluorenylmethoxycarbonyl, dichloroacetyl, o-nitrophenylsulfenyl, 2,2,2-trichloroethoxycarbonyl, bromo-t-butoxycarbonyl, phenoxyacetyl; non-acyl protective groups such as triloweralkylsilyl, for example, trimethylsilyl and t-butyldimethylsilyl are also of interest.

The following radicals, according to the foregoing definition of acyl (radicals R.sup.1 and R.sup.2 of Structure II, above), are preferred: formyl, propionyl, butyryl, chloroacetyl, methoxyacetyl, aminoacetyl, methoxycarbonyl, ethoxycarbonyl, methylcarbamoyl, ethylcarbamoyl, phenylthiocarbonyl, 3-aminopropionyl, 4-aminobutyryl, N-methylaminoacetyl, N,N-dimethylaminoacetyl, N,N,N-trimethylaminoacetyl, 3-(N,N-dimethyl)aminopropionyl, 3-(N,N,N-trimethyl)amino propionyl, N,N,N-triethylaminoacetyl, pyridiniumacetyl, guanylthioacetyl, guanidinoacetyl, 3-guanidinopropionyl, N.sup.3 -methylguanidinopropionyl, hydroxyacetyl, 3-hydroxypropionyl, acryloyl, propynoyl, malonyl, phenoxycarbonyl, amidinoacetyl, acetamidinoacetyl, amidinopropionyl, acetamidinopropionyl, guanylureidoacetyl, guanylcarbamoyl, carboxymethylaminoacetyl, sulfoacetylaminoacetyl, phosphonoacetylaminoacetyl, N.sup.3 -dimethylaminoacetamidinopropionyl, ureidocarbonyl, dimethylaminoguanylthioacetyl, 3-(1-methyl-4-pyridinium)propionyl, 3-(5-aminoimidazol-1-yl)propionyl, 3-methyl-1-imidazoliumacetyl, 3-sydnonylacetyl, o-aminomethylbenzoyl, o-aminobenzoyl, ##STR12##

An especially preferred class of acyl radicals (R.sup.1 and R.sup.2 of structure II, above) are terminally substituted acyls wherein the substituent is a basic group such as substituted and unsubstituted: amino, amidino, guanidino, guanyl and nitrogen-containing mono- and bicyclic heterocycles (aromatic and non-aromatic) wherein the hetero atom or atoms, in addition to nitrogen, are selected from oxygen and sulphur. Such preferred substituted acyls may be represented by the following formula: ##STR13## wherein m, and n are integers selected from 0 to 5; A is O, NR' (R' is hydrogen or loweralkyl having 1-6 carbon atoms), S or A represents a single bond; and Y is selected from the following group:

(1) amino or substituted amino:

--N(R).sub.2 and --N.sup.+ (R).sub.3 wherein the values for R are independently selected from: hydrogen; N(R').sub.2 (R' is hydrogen or loweralkyl having 1-6 carbon atoms); loweralkyl and loweralkoxyl having from 1 to 6 carbon atoms; loweralkoxyloweralkyl wherein the alkoxyl moiety comprises 1 to 6 carbon atoms and the alkyl moiety comprises 2-6 carbon atoms; cycloalkyl and cycloalkylalkyl wherein the cycloalkyl moiety comprises 3-6 carbon atoms and the alkyl moiety comprises 1-3 carbon atoms; two R groups may be joined together with the N atom to which they are attached to form a ring having 3-6 atoms.

(2) amidino and substituted amidino: ##STR14## wherein the value of R is independently selected from the group consisting of: hydrogen; N(R').sub.2 (R' is hydrogen or loweralkyl having 1-6 carbon atoms); loweralkyl and loweralkoxyl having from 1 to 6 carbon atoms; loweralkoxyloweralkyl wherein the alkoxyl moiety comprises 1 to 6 carbon atoms and the alkyl moiety comprises 2 to 6 carbon atoms (when the loweralkoxyloweralkyl radical is attached to carbon the alkyl moiety comprises 1 to 6 carbon atoms); cycloalkyl and cycloalkylalkyl wherein the alkyl moiety comprises 1 to 3 carbon atoms; two R groups may be joined together with the atoms to which they are attached to form a ring having 3 to 6 atoms;

(3) guanidino and substituted guanidino: ##STR15## wherein R is as defined in 2 (above).

(4) guanyl and substituted guanyl: ##STR16## wherein R is as defined in 2 (above).

(5) nitrogen-containing mono- and bicylic heterocycles (aromatic and non-aromatic) having 4 to 10 nuclear atoms wherein the hetero atom or atoms, in addition to nitrogen, are selected from oxygen and sulphur. Such heterocycles are representatively illustrated by the following list of radicals (R' is H or loweralkyl having 1-6 carbon atom): ##STR17##

The following specific acyl radicals falling within this class are additionally representative and are preferred: ##STR18##

However, it is to be understood that any acyl radical may be employed in the practice of the invention and is to be considered within the scope of the invention.

The N-acylated intermediate (1, above) is prepared by treating thienamycin (I) with an acylating agent, for example, an acyl halide or acyl anhydride such as an aliphatic, aromatic, heterocyclic, araliphatic or heterocyclic aliphatic carboxylic acid halide or anhydride. Other acylating agents may also be employed, for example, mixed carboxylic acid anhydrides and particularly lower alkyl esters of mixed carboxylic-carbonic anhydrides; also, carboxylic acids in the presence of a carbodiimide such as 1,3-dicyclohexylcarbodiimide, and an activated ester of a carboxylic acid such as the p-nitrophenyl ester.

The acylation reaction may be conducted at a temperature in the range of from about -20.degree. C. to about 100.degree. C. but is preferably conducted at a temperature in the range of from -8.degree. C. to 25.degree. C. Any solvent in which the reactants are soluble and substantially inert may be employed, for example, polar solvents such as water, alcohols and polar organic solvents in general such as dimethylformamide (DMF), hexamethyl phosphoramide (HMPA), acetone, dioxane, tetrahydrofuran (THF), acetonitrile, heterocyclic amines such as pyridine, ethylacetate, aqueous mixtures of the above, as well as halogenated solvents such as methylene chloride and chloroform. The reaction is conducted for a period of time of from about five minutes to a maximum of three hours, but in general, a reaction time of about 0.5 to about one hour is sufficient. The following equation illustrates this process employing a carboxylic acid halide; however, it is to be understood that by substituting a carboxylic acid anhydride or other functionally equivalent acylating agent similar products may be obtained. ##STR19##

Generally when the above-described acylating reaction employs an acid halide (suitable halides are chloro, iodo or bromo) or anhydride the reaction is conducted in water or an aqueous mixture of a polar organic solvent such as acetone, dioxane, THF, DMF, acetonitrile or the like in the presence of a suitable acceptor base such as NaHCO.sub.3, MgO, NaOH, K.sub.2 HPO.sub.4 and the like.

In carrying out the reactions described herein, it is generally not necessary to protect the 2-carboxy group or the 1'-hydroxy group; however, in cases where the acylating reagent is exceedingly water sensitive it is sometimes advantageous to perform the acylation in a non aqueous solvent system. Triorganosilyl (or tin) derivatives of thienamycin are suitable for this purpose. Silylation of thienamycin proceeds rapidly to give the tris-triorganosilyl derivative, for example tris-trimethylsilyl thienamycin, Th(TMS).sub.3 : ##STR20## Such derivatives, which are readily soluble in organic solvents, are conveniently prepared by treating Thienamycin with an excess of hexamethyldisilazane and a stoichiometric amount of trimethylchlorosilane at 25.degree. C. with vigorous stirring under a N.sub.2 atmosphere. The resulting NH.sub.4 Cl is removed by centrifugation and the solvent is removed by evaporation to provide the desired silyl derivative.

In the generic representation of the compounds of the present invention (II, above), the radical represented by --COXR, is, inter alia, all radicals known to be effective as pharmaceutically acceptable ester, anhydride (R is acyl) and amide radicals in the bicyclic .beta.-lactam antibiotic art, such as the cephalosporins and penicillins and the nuclear analogues thereof.

Suitable radicals (R) include conventional protecting or carboxyl blocking groups. The term "blocking group" as utilized herein is employed in the same manner and in accordance with the teaching of U.S. Pat. No. 3,697,515, which is incorporated herein by reference. Pharmaceutically acceptable Thienamycin derivatives of the present invention falling in this class are given below. Suitable blocking esters thus include those selected from the following list which is representative and not intended to be an exhaustive list of possible ester groups, wherein X=O and R is given:

(i) R=CR.sup.a R.sup.b R.sup.c wherein at least one of R.sup.a, R.sup.b and R.sup.c is an electron-donor, e.g., p-methoxyphenyl, 2,4,6-trimethylphenyl, 9-anthryl, methoxy, CH.sub.2 SCH.sub.3, tetrahydrofur-2-yl, tetrahydropyran-2-yl or fur-2-yl. The remaining R.sup.a, R.sup.b and R.sup.c groups may be hydrogen or organic substituting groups. Suitable ester groups of this type include L-methoxybenzyloxycarbonyl and 2,4,6-trimethylbenzyloxycarbonyl.

(ii) R=CR.sup.a R.sup.b R.sup.c wherein at least one of R.sup.a, R.sup.b and R.sup.c is an electron-attracting group, e.g., benzoyl, p-nitrophenyl, 4-pyridyl, trichloromethyl, tribromomethyl, iodomethyl, cyanomethyl, ethoxycarbonylmethyl, arylsulphonylmethyl, 2-dimethylsulphoniumethyl, o-nitrophenyl or cyano. Suitable esters of this type include benzoylmethoxycarbonyl, p-nitrobenzyloxycarbonyl, 4-pyridylmethoxycarbonyl, 2,2,2-trichloromethoxycarbonyl and 2,2,2-tribromoethoxycarbonyl.

(iii) R=CR.sup.a R.sup.b R.sup.c wherein at least two of R.sup.a, R.sup.b and R.sup.c are hydrocarbon such as alkyl, e.g. methyl or ethyl, or aryl e.g., phenyl and the remaining R.sup.a, R.sup.b and R.sup.c group, if there is one, is hydrogen. Suitable esters of this type include t-butyloxycarbonyl, t-amyloxycarbonyl, diphenylmethoxycarbonyl and triphenylmethoxycarbonyl.

(iv) R=R.sup.d, wherein R.sup.d is adamantyl, 2-benzyloxyphenyl, 4-methylthiophenyl or tetrahydropyran-2-yl.

Silyl esters, under this category of blocking groups, may conveniently be prepared from a halosilane or a silazane of the formula: R.sup.4.sub.3 SiX'; R.sup.4.sub.2 Six'.sub.2 ; R.sup.4.sub.3 Si.NR.sup.4.sub.2 ; R.sup.4.sub.3 Si.NH.COR.sup.4 ; R.sup.4.sub.3 Si.NH.CO.NH.SiR.sup.4.sub.3 ; R.sup.4 NH.CO.NR.sup.4.SiR.sup.4.sub.3 ; or R.sup.4 C(OSiR.sup.4.sub.3); HN(SiR.sup.4.sub.3).sub.2 wherein X' is a halogen such as chloro or bromo and the various groups R.sup.4, which can be the same of different, represent hydrogen atoms or alkyl, e.g., methyl, ethyl, n-propyl, iso-propyl; aryl; e.g., phenyl; or aralkyl, e.g., benzyl, groups.

More generally stated, pharmaceutically acceptable carboxyl derivatives of the present invention are those derived by reacting I or the N-acylated thienamycin (1) with alcohols, phenols, mercaptans, thiophenols, acylating reagents and the like which yield compounds II (above) which are bio-labile to conversion to the N-acyl thienamycin (1) which are also active as antibiotic or to thienamycin itself.

For example, esters and amides of interest are the compounds of the formula II (above) having the following group at the 2-position: --COXR wherein X is oxygen, sulfur or NR' (R' is as defined above), and R is alkyl having 1-10 carbon atoms, straight or branched, such as methyl, ethyl, t-butyl, pentyl, decyl and the like; carbonylmethyl including phenacyl, p-bromophenacyl, p-t-butylphenacyl, acetoxyacetylmethyl, pivaloxyacetylmethyl, carboxymethyl and its alkyl and aryl esters, .alpha.-carboxy-.alpha.-isopropyl; aminoalkyl including 2-methylaminoethyl, 2-diethylaminoethyl, 2-acetamidoethyl, phthalimidomethyl, succinimidomethyl; alkoxyalkyl wherein the alkoxy portion has 1-10 and preferably 1-6 carbon atoms, but can be branched, straight or cyclic, and the alkyl portion has 1-6 carbon atoms, such as methoxymethyl, ethoxymethyl, isopropoxymethyl, decyloxymethyl, ethoxypropyl, decyloxypentyl, cyclohexyloxymethyl and the like; alkanoyloxyalkyl wherein the alkanoyloxy portion is straight or branched and has 1-6 carbon atoms and the alkyl portion has 16 carbon atoms, such as acetoxymethyl, pivaloyloxymethyl, acetoxyethyl, propionyloxyethyl, acetoxypropyl, and the like; haloalkyl wherein halo is chloro, bromo, fluoro, or iodo, and the alkyl portion is straight or branched having 1-6 carbon atoms, e.g., 2,2,2-trichloroethyl, trifluoroethyl, 2-bromopropyl, diiodomethyl, 2-chloroethyl, 2-bromoethyl, and the like; alkenyl having 1-10 carbon atoms, either straight or branched, e.g., allyl, 2-propenyl, 3-butenyl, 4-butenyl, 4-pentenyl, 2-butenyl, 3-pentenyl, 3-methyl-3-butenyl, metallyl, 1,4-cyclohexadien-1-yl-methyl, and the like alkynyl having 1-10 carbon atoms, either straight or branched, e.g., 3-pentynyl, propargyl, ethynyl, 3-butyn-1-yl, and the like; alkanoyl, either straight or branched, having 1-10 carbon atoms, such as pivaloyl, acetyl, propionyl, and the like; aralkyl or heteroaralkyl wherein alkyl has 1-3 carbon atoms, and hetero means 1-4 hetero atoms being selected from the group consisting of O, S, or N, such as benzyl, benzhydryl, and substituted benzyl, benzhydryl, or e.g., benzyl or benzhydryl substituted with 1-3 substituents such as benzyl, phenoxy, halo, loweralkyl, loweralkanoyloxy of 1-5 carbon atoms, lower alkoxy, hydroxy, nitro, blocked carboxy, or combinations thereof, e.g., p-chlorobenzyl, o-nitrobenzyl, 3,5-dinitrobenzyl, p-methoxybenzyl, m-benzoylbenzyl, p-t-butylbenzyl, m-phenoxybenzyl, p-benzoylbenzyl, p-nitrobenzyl, 3,5-dichloro-4-hydroxybenzyl, p-methoxycarbonylbenzyl, p-methoxybenzhydryl, p-carboxybenzyl, the latter being either the free acid, ester or the sodium salt, 2,4,6-trimethylbenzyl, p-pivaloyloxybenzyl, p-t-butoxycarbonyl benzyl, p-methylbenzyl, p-benzoyloxybenzyl, p-acetoxybenzyl, p-2-ethylhexanoylbenzoyl, p-ethoxycarbonylbenzyl, p-benzoylthiobenzyl, p-benzamidobenzyl, o-pivaloxyloxybenzyl, m-pivaloyloxybenzyl, p-isopropoxybenzyl, p-t-butoxybenzyl, as well as cyclic analogues thereof, 2,2-dimethyl-5-coumaranmethyl, 5-indanylmethyl, p-trimethylsilylbenzyl, 3,5-bis-t-butoxy-4-hydroxybenzyl; 2-thienylmethyl, 2-furylmethyl, 3-t-butyl-5-isothiazolmethyl, 6-pivaloyloxy-3-pyridazinylethyl, 5-phenylthio-1-tetrazolylmethyl, or the like (the use of the terms lower alkyl or lower alkoxy in this context means 1-4 carbon atom chain); or phthalidyl; or phenylethyl, 2-(p-methylphenyl)ethyl, and the arylthioalkyl analogues, aryloxyalkyl wherein aryl is preferably a phenyl ring having 0-3 substituents, preferably 0 or 1 substituents in the ortho or para positions and alkyl is 1-6 carbon atoms, e.g., (4-methoxy)phenoxymethyl, phenoxymethyl, (4-chloro)phenoxymethyl, (4-nitro)phenoxymethyl, (4-benzyloxy)phenoxymethyl, (4-methyl)phenoxymethyl, (4-benzyloxy)phenoxymethyl, (4-methyl)phenoxymethyl, (2-methoxy)phenoxymethyl, (1-phenoxy)ethyl, (4-amino)phenoxymethyl, (4-methoxy)phenylthiomethyl, (4-chloro)phenylthiomethyl, phenylthioethyl, aryl wherein aryl is phenyl, 5-indanyl, or substituted phenyl having 0-3 substituents, preferably 0 or 1 substituent in the ortho or para position, e.g., (4-methyl)phenyl, (4-hydroxy)phenyl, (4-t-butyl)phenyl, p-nitrophenyl, 3,5-dinitrophenyl or p-carboxyphenyl, the latter having either the free acid or the sodium salt form; aralkenyl wherein aryl is phenyl and alkenyl has 1-6 carbon atoms, such as 3-phenyl-2-propenyl; aralkoxyalkyl wherein aralkoxy is benzyloxy, and alkyl has 1-3 carbon atoms, such as benzyloxymethyl, (4-nitro)benzyloxymethyl, (4-chloro)benzyloxymethyl; alkylthioalkyl wherein the alkylthio portion has 1-10 and preferably 1-6 carbon atoms, but can be branched, straight, or cyclic, and the alkyl portion has 1-6 carbon atoms, such as methylthioethyl, ethylthioethyl, cyclohexylthiomethyl, decylthiobutyl, methylthiopropyl, isopropylthioethyl, methylthiobutyl and the like; alkoxycarbonyloxymethyl, dialkylaminoacetoxymethyl, and alkanoylamidomethyl, wherein the alkyl moieties of the last three mentioned radicals each comprise 1-6 carbon atoms.

In addition to the esters (and thio esters) listed above, amides are also embraced by the present invention, i.e., wherein X is the ##STR21## Representative of such amides, Th-CONR'R, are those wherein R' is selected from the group consisting of hydrogen, methyl, ethyl, phenyl, p-methoxyphenyl, benzyl, carboxymethyl, methylthioethyl, and heteroaryl; also embraced by --COXR are anhydrides wherein R is benzyloxycarbonyl, ethoxycarbonyl, benzoyl, and pivaloyl.

Particularly preferred esters are those wherein X is oxygen, sulphur or NR' (R' is hydrogen or lower alkyl having 1-6 carbon atoms) and R is aralkyl, aryloxyalkyl, aralkoxyalkyl, alkylthioalkyl, haloalkyl and alkenyl.

The most preferred compounds of the present invention are those wherein (relative to structure II, above) X is oxygen, and R is selected from the group consisting of: lower alkyl, lower alkenyl, such as methallyl, 3-methylbutenyl, 3-butenyl and the like; methylthio ethyl, benzyl and substituted benzyl, such as p-t-butylbenzyl, m-phenoxybenzyl, p-pivaloyloxybenzyl, p-nitrobenzyl and the like; pivaloyloxymethyl, 3-phthalidyl, acetoxymethyl, propionyloxymethyl, acetylthiomethyl, pivaloylthiomethyl, allyl, 4-pentenyl, 2-butenyl, 3-methyl-2-butenyl, phenacyl, acetoxyacetylmethyl, pivaloyacetylmethyl, diethylaminoethyl, dimethylaminoethyl, methoxymethyl, p-acetoxybenzyl, p-pivaloyloxybenzyl, p-isopropoxybenzyl, 5-indanylmethyl, 5-indanyl, benzyloxymethyl, etylthioethyl, methylthiopropyl, methoxycarbonyloxymethyl, ethoxycarbonyloxymethyl, dimethylaminoacetoxymethyl, crotonolacton-3-yl, and acetamidomethyl.

The preferred N-blocking groups for the starting material 1 are: carbobenzyloxy, ring-substituted carbobenzyloxy such as o- and p-nitrocarbobenzyloxy, p-methoxycarbobenzyloxy, chloroacetyl, bromoacetyl, phenylacetyl, t-butoxycarbonyl, trifluoroacetyl, bromoethoxycarbonyl, 9-fluorenylmethoxycarbonyl, dichloroacetyl, o-nitrophenylsulfenyl, 2,2,2-trichloroethoxycarbonyl, bromo-t-butoxycarbonyl, phenoxyacetyl; non-acyl protective groups such as alkylidenes, for example, benzylidene and salicylidene are also of interest.

As noted above, the compounds of the present invention (II,above) are prepared according to the following scheme: ##STR22## wherein all symbolism is as previously defined and the N-acylated starting material 1 is fully described above and in co-pending U.S. patent application Ser. No. 634,291, filed Nov. 21, 1975, now abandoned in favor of U.S. patent application Ser. No. 733,653, filed Oct. 18, 1976, now abandoned in favor of U.S. patent application Ser. No. 861,247, filed Dec. 16, 1977, now abandoned in favor of U.S. patent application Ser. No. 160,718, filed June 18, 1980, now abandoned in favor of U.S. patent application Ser. No. 321,496, filed Nov. 16, 1981; these applications are incorporated herein by reference for their disclosure relative to the preparation of 1.

In general, the transformation (1.fwdarw.II) of the present invention is accomplished by conventional procedures known in the art. Such procedures include:

(1) Reaction of 1, with a diazoalkane such as diazomethane, phenyldiazomethane, diphenyldiazomethane, and the like, in a solvent such as dioxane, ethylacetate, acetonitrile and the like at a temperature of from 0.degree. C. to reflux for from a few minutes to 2 hours.

(2) Reaction of an alkali metal salt of 1 with an activated alkyl halide such as methyl iodide, benzyl bromide, or m-phenoxybenzyl bromide, p-t-butylbenzyl bromide, pivaloyloxymethyl chloride, and the like. Suitable reaction conditions include solvents such as hexamethylphosphoramide and the like at a temperature of from 0.degree. C., to 60.degree. C., for from a few minutes to 4 hours.

(3) Reaction of 1 with an alcohol such as methanol, ethanol, benzyl alcohol, and the like. This reaction may be conducted in the presence of a carbodiimide condensing agent such as dicyclohexylcarbodiimide or the like. Suitable solvents, at a temperature of from 0.degree. C. to reflux for from 15 minutes to 18 hours, include CHCl.sub.3, CH.sub.3 CH, CH.sub.2 Cl.sub.2 and the like.

(4) Reaction of an N-acylated acid anhydride of 1 prepared by reacting the free acid 1 with an acid chloride such as ethylchloroformate, benzylchloroformate and the like, with an alcohol such as those listed in (3) under the same conditions of reaction as given above for (3). The anhydride is prepared by reacting III and the acid chloride in a solvent such as tetrahydrofuran (THF), CH.sub.2 Cl.sub.2 and the like at a temperature of from 25.degree. C., to reflux for from 15 minutes to 10 hours.

(5) Reaction of labile esters of thienamycin, I, such as trimethylsilyl ester, dimethyl-t-butylsilyl ester or the like with RX' whereix X' is halogen such as bromo and chloro and R is as defined, in a solvent such as THF, CH.sub.2 Cl.sub.2 and the like at a temperature of from 0.degree. C. to reflux for from 15 minutes to 16 hours. For example according to the following scheme: ##STR23## wherein TMS is triorganosilyl such as trimethylsilyl and all other symbolism is as previously defined.

The amides of the present invention are most conveniently prepared by reacting the acid anhydride of II (X.dbd.O, R.dbd.acyl) with ammonia or with the amine of choice, e.g, the alkyl-, dialkyl-, aralkyl- or hetero cyclic amines listed above.

The above-recited schemes of esterification are well known in the related bicyclic .beta.-lactam antibiotic art and indeed in all of general organic synthesis and it is to be noted that there is no undue criticality of reaction parameters in the preparation of the N-acylated, carboxyl derivatives (II) of the present invention.

It is again to be noted that the compounds of the present invention may be prepared by establishing the N-acyl substituent upon the preformed ester: ##STR24##

The compounds of the present invention are also useful for the preparation of other thienamycin derivatives 3 and 4, which are useful as antibiotics: ##STR25## wherein R.sup.3 is an ether (R.sup.3 is R or is defined by acyl radicals R.sup.1 and R.sup.2 wherein the carbonyl moiety ##STR26## or more generally ##STR27## is deleted) or an ester (R.sup.3 is defined by acyl radicals R.sup.1 and R.sup.2) moiety (see above definition of R.sup.1, R.sup.2 and R relative to the compounds of the present invention II); and all other symbolism is as previously defined. The compounds 3 are fully described in co-pending U.S. patent application Ser. No. 634,006, filed Nov. 21, 1975, now abandoned in favor of U.S. patent application Ser. No. 733,655, filed Oct. 18, 1976, now abandoned in favor of U.S. patent application Ser. No. 861,234, filed Dec. 12, 1977, now U.S. Pat. No. 4,208,330, issued June 17, 1980. These applications are incorporated herein by reference as they relate to the utilization of the compounds of the present invention as intermediates in the preparation of 3 above. The compounds 4 are fully described in co-pending U.S. patent application Ser. No. 634,294, filed Nov. 21, 1975, now abandoned in favor of U.S. patent application Ser. No. 733,652, filed Oct. 18, 1976, now abandoned in favor of U.S. patent application Ser. No. 861,246, filed Dec. 16, 1977, now U.S. Pat. No. 4,226,870, issued Oct. 17, 1981. These applications are incorporated by reference as they relate to the utilization of the compounds of the present invention in the preparation of 4, above.

Compounds 3 and 4 are conveniently prepared from II, above, according to the following scheme: ##STR28## wherein X is preferably oxygen and either R.sup.1 or R.sup.2 is hydrogen. When it is desired to prepare 3 R is chosen from one of the above-enumerated, carboxyl blocking or protective groups and R.sup.1 and R.sup.2 are chosen from easily removable N-blocking or protective group such as carbobenzyloxy, ring-substituted carbobenzyloxy such as o- and p-nitrocarbobenzyloxy, p-methoxycarbobenzyloxy, chloroacetyl, bromoacetyl, phenylacetyl, t-butoxycarbonyl, trifluoroacetyl, bromoethoxycarbonyl, 9-fluorenylmethoxycarbonyl, dichloroacetyl, o-nitrophenylsulfenyl, 2,2,2-trichloroethoxycarbonyl, bromo-t-butoxycarbonyl, phenoxyacetyl; non-acyl protective groups such as triloweralkylsilyl, for example, trimethylsilyl and t-butyldimethylsilyl are also of interest. In this connection, preferred carboxyl blocking groups R include the sub-generic groups defined above as aralkyl, haloalkyl, alkanoyloxyalkyl, alkoxyalkyl, alkenyl, substituted alkyl, or aralkoxyalkyl, and also including alkylsilyl, wherein alkyl has 1-10 carbon atoms. For example, suitable R "blocking groups" include benzyl, phenacyl, p-nitrobenzyl, methoxymethyl, trichloroethyl, trimethylsilyl, tributyltin, p-methoxybenzyl, benzhydryl. These blocking groups are preferred since they are generally recognized as easily-removable blocking groups in the cephalosporin and penicillin art.

Deblocking 4.fwdarw.3 is preferably conducted as a single step according to procedures well known in the art; however, frequently, depending on the identity of R.sup.1 /R.sup.2 and R, it is desirable to deblock in separate steps. The most commonly preferred deblocking procedure is hydrogenation; wherein the species to be deblocked, in a solvent such as a lower alkanol, is hydrogenated in the presence of hydrogenation catalysts such as Pt, Pd or oxides thereof.

In general, the compounds 3 and 4 are prepared by any of a variety of well-known esterification or etherification reactions (II.fwdarw.4) upon the secondary alcoholic group of thienamycin in its protected form, II. Such procedures (II.fwdarw.4) include:

(1) For the preparation of ether embodiments, the acid catalized reaction of II with a diazoalkane such as diazomethane, phenyldiazomethane, diphenyldiazomethane and the like in an inert solvent such as dioxane, tetrahydrofuran (THF), halohydrocarbons such as CH.sub.2 Cl.sub.2, ethylacetate and the like in the presence of a catalytic amount of a strong acid or Lewis acid such as toluenesulfonic acid, trifluoroacetic acid, fluoboric acid, boron trifluoride, and the like at a temperature of from -78.degree. C. to 25.degree. C. for from a few minutes to 2 hours.

(2) For the preparation of ether embodiments, the reaction of II with an alkylating agent such as active halides for example methyliodide, benzylbromide, m-phenoxybenzylbromide and the like and alkylsulphates such as dimethylsulphate, diethylsulphate, methylfluorosulphonate and the like in the presence of a strong base capable of forming the alcoholate anion of II. Suitable bases include alkali and alkaline earth metal oxides and hydrous oxides, alkali metal alkoxides such as potassium tertiary-butoxide, tertiary amines such as triethylamine, alkali metal alkyls and aryls such as phenyllithium, and alkali metal amides such as sodium amide. Suitable solvents include any inert anhydrous solvent such as t-butanol, dimethylformamide (DMF), THF, hexamethylphosphoramide (HMPA) dioxane and the like at a temperature of from -78.degree. C. to 25.degree. C., for from a few minutes to 4 hours.

(3) For the preparation of ester embodiments, the reaction of II with any of the above-listed acyl radicals in their acid form. This reaction may be conducted in the presence of a carbodiimide condensing agent such as cyclohexylcarbodiimide or the like. Suitable solvents include any inert solvent such as CHCl.sub.3, CH.sub.2 Cl.sub.2, DMF, HMPA, acetone, dioxane and the like at a temperature of from 0.degree. C. to 60.degree. C. for from 15 minutes to 12 hours.

(4) For the preparation of ester embodiments, the reaction of II with an acyl halide or an acid anhydride wherein the acyl moiety is described above. Generally, when the above-described acylating reaction employs an acid halide (suitable halides are chloro, iodo or bromo or acid anhydride) the reaction is conducted in an anhydrous organic solvent such as acetone, dioxane, methylenechloride, chloroform, DMF, or the like in the presence of a suitable acceptor base such as NaHCO.sub.3, MgO, triethylene, pyridine, and the like at a temperature of from 0.degree. C. to 40.degree. C. for from 1 to 4 hours.

Suitable acyl halides and anhydrides include: acetic anhydride, bromoacetic anhydride, propionic anhydride, benzoylchloride, phenylacetyl chloride, azidoacetyl chloride, 2-thienylacetyl chloride, 2-, 3- and 4-nicotinyl chloride, p-nitrobenzoyl chloride, 2,6-dimethoxybenzoyl chloride, 4-guanidinophenylacetyl chloride, hydrochloride, methanesulfonyl chloride, dibenzylphosphorochloridate, dimethylthiophosphorochloridate, 2-furoyl ethyl, 2-furoyl ethyl carbonic anhydride, methylchloroformate, bis-(p-nitrobenzyl)phosphorochloridate and the like.

(5) For the preparation of ester embodiments, the reaction of II with a suitably substituted ketene or isocyanate such as ketene, dimethyl ketene, methylisocyanate, methylisothiocyanate, chlorosulfonyl isocyanate and the like. Suitable solvents include dioxane, tetrahydrofuran, chloroform and the like at a temperature of from -70.degree. C. to 60.degree. C. for from 15 minutes to 18 hours.

The compounds of the present invention are valuable antibiotics active against various gram-positive and gram-negative bacteria and, accordingly, find utility in human and veterinary medicine and in inanimate systems. The compounds of this invention can therefore be used as antibacterial drugs for treating infections caused by gram-positive or gram-negative bacteria, for example against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Bacillus substilis, Salmonella typhosa, Pseudomonas and Bacterium proteus. The antibacterial compounds of the invention may further be utilized as additives to animal feedingstuffs, for preserving foodstuffs and as disinfectants. For example, they may be employed in aqueous compositions in concentrations ranging from 0.1 to 100 parts of antibiotic per million parts of solution in order to destroy and inhibit the growth of harmful bacteria on medical and dental equipment and as bactericides in industrial applications, for example in waterbased paints and in the white water of paper mills to inhibit the growth of harmful bacteria.

The products of this invention may be used alone or in combination as an active ingredient in any one of a variety of pharmaceutical preparations: These antibiotics and their corresponding pharmaceutically acceptable salt ester and amide derivatives may be employed in capsule form or as tablets, powders or liquid solutions or as suspensions or elixirs. They may be administered orally, intravenously or intramuscularly. Such pharmaceutically acceptable forms are prepared according to procedures well-known in the art.

The compositions are preferably presented in a form suitable for absorption by the gastro-intestinal tract. Tablets and capsules for oral administration may be in unit dose presentation form, and may contain conventional excipients such as binding agents, for example, syrup, acacia, gelatin, sorbitol, tragacanth, or polyvinylpyrrolidone; fillers for example, lactose, sugar, maizestarch, calcium phosphate, sorbitol or glycine; lubricants, for example, magnesium stearate, talc, polyethylene glycol, silica; disintegrants, for example, potato starch or acceptable wetting agents such as sodium lauryl sulphate. The tablets may be coated according to methods well known in the art. Oral liquid preparation may be in the form of aqueous or oily suspension, solution, emulsions, syrups, elixirs, etc. or may be presented as a dry product, for reconstitution with water or other suitable vehicles before use. Such liquid preparations may contain conventional additives such as suspending agents, for example, sorbitol syrup, methylcellulose, glucose/sugar syrup, gelatin, hydroxyethylcellulose, carboxymethyl cellulose, aluminum stearate gel or hydrogenated edible oils, for example almond oil, fractionated coconut oil, oily esters, propylene glycol, or ethyl alcohol; preservatives, for example methyl or propyl p-hydroxybenzoates or sorbic acid. Suppositories will contain conventional suppository bases, e.g., cocoa butter or other glycerides.

Compositions for injection may be presented in unit dose form in ampules or in multidose containers with an added preservative. The compositions may take such forms as suspensions, solutions, emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents. Alternatively, the active ingredient may be in powder form for reconstitution with a suitable vehicle, e.g., sterile, pyrogen-free water, before use.

The compositions may also be prepared in suitable forms for absorption through the mucous membranes of the nose and throat or bronchial tissues and may conveniently take the form of powder or liquid sprays or inhalants, lozenges, throat paints, etc. For medication of the eyes or ears, the preparations may be presented as individual capsules, in liquid or semi-solid form, or may be used as drops. Topical applications may be formulated in hydrophobic or hydrophilic bases as ointments, creams, lotions, paints, or powders.

Also, in addition to a carrier, the instant compositions may include other ingredients such as stabilizers, binders, antioxidants, preservatives, lubricators, suspending agents, viscosity agents or flavoring agents and the like. In addition, there may also be included in the composition other active ingredients to provide a broader spectrum of antibiotic activity.

For veterinary medicine the composition may, for example, be formulated as an intramammary preparation in either long acting or quick-release bases.

The dosage to be administered depends to a large extent upon the condition of the subject being treated and the weight of the host, the route and frequency of administration, the parenteral route being preferred for generalized infections and the oral route for intestinal infections. In general, a daily oral dosage consists of from about 15 to about 600 mg. of active ingredient per kg. of body weight of the subject in one or more applications per day. A preferred daily dosage for adult humans lies in the range of from about 80 to 120 mg. of active ingredient per kg. of body weight.

The instant compositions may be administered in several unit dosage forms as, for example, in solid or liquid orally ingestible dosage form. The compositions per unit dosage, whether liquid or solid may contain from 0.1 to about 99% of active material, the preferred range being from about 10-60%. The compositions will generally contain from about 15 mg. to about 1500 mg. of the active ingredient; however, in general, it is preferable to employ a dosage amount in the range of from about 250 mg. to 1000 mg. In parenteral administration the unit dosage is usually the pure compounds in a slightly acidified sterile water solution or as the form of a soluble powder intended for solution.

In the following Examples, which further illustrate but do not limit the product, process, compositional or method of treatment aspects of the present invention, the compounds of the present invention will be designated by the previously introduced symbol: ##STR29## wherein the three functional groups are illustrated.

EXAMPLE 1 ##STR30##

Preparation of N-(O-formyl)-D-mandeloylthienamycin Sodium Salt

To thienamycin (40 mg.) in 10 ml. water is added successively at 0.degree. C., 124 mg. NaHCO.sub.3, 8 ml. dioxane and then with stirring 1.2 equivalent N-(O-formyl)-1-mandelolyl chloride over a period of one minute. After six minutes total reaction time, the mixture is extracted three times with cold ethyl ether. Electrophoresis of an aqueous portion (0.05 M, pH 7, aqueous phosphate buffer, 50 V/cm., 20 minutes) shows 67% conversion to desired product. The pH is adjusted to 2.2 with 1 M H.sub.3 PO.sub.4 solution and the solution is extracted three times with ethyl acetate. The ethyl acetate (EtOAc) solution is dried over MgSO.sub.4 and extracted twice with two equivalents of NaHCO.sub.3 solution. The aqueous extract, lyophilized, contains 164 optical density units (ODU), at 302 nm by uv analysis at pH 7.0, of which 95% is extinguished after treatment with hydroxylamine for one hour. The yield is 53%. Electrophoresis as before shows one spot by bioautograph, 4 cm towards the anode. NMR (.delta.D.sub.2 O) 1.30 (d, J=6 Hz, CH.sub.3 CH); 2.8-3.7 (m, CH.sub.2), 4.0-4.5 (m, CH .beta.-lactam), 4.73, HDO; 5.97 (s, C.sub.6 H.sub.5 CHOCHO), 7.53 (s, C.sub.6 H.sub.5), 8.30 (s, C.sub.6 H.sub.5 CHOCHO).

EXAMPLE 2 ##STR31##

Preparation of N-D-Mandeloylthienamycin Sodium Salt

The title compound is made following the procedure of Example 1, but before EtOAc extraction the aqueous extract was allowed to stand at 25.degree. C. for one hour. Electrophoresis (50 V/cm., 20 min., pH 7, aqueous phosphate, 0.05 M) shows one spot by bioautograph, 4 cm. towards the anode, NMR (.delta., D.sub.2 O) 1.50 (d, J=6 Hz, CH.sub.3 CH); 2.8-3.8 (m, CH.sub.2), 4.2-4.6 (m, CH .beta.-lactam); 4.96 (s, HDO); ##STR32##

EXAMPLE 3

N-Propionylthienamycin Sodium Salt

To Thienamycin, (25 mg. in 6 ml. water at 0.degree. C.) is successively added 38.6 mg. NaHCO.sub.3, 5 ml. dioxane and then with stirring one equivalent of propionic anhydride over a period of 3 min. After 10 min., the mixture is extracted three times with cold ethyl ether. Electrophoresis of the aqueous (aq.) extract (0.05 M, pH 7, phosphate buffer, 50 V/cm., 20 min.) shows no free Thienamycin present. The aq. extract is adjusted to pH 6.8 and contains 600 ODU at 302 nm by uv analysis which is 95% extinguished after treatment with hydroxylamine for one hour. NMR (.delta., D.sub.2 O) 1.42 (m, CH.sub.2 CH.sub.3, CH.sub.3 CH); 2.48 (q, CH.sub.2 CH.sub.3); 2.86-2.90 (m, CH.sub.2), 4.30-4.70 (m, CH .beta.-lactam), 4.86 (HDO).

EXAMPLE 4

N-(Methoxyacetyl)thienamycin Sodium Salt

To thienamycin, (55 mg. in 6 ml. water at 0.degree. C.), is added successively 68 mg. NaHCO.sub.3, 6 ml. dioxane and with stirring 1.1 equivalents methoxyacetylchloride over a period of 1.5 minutes. The mixture is stirred an additional 10 minutes. The mixture is extracted three times with cold ethyl ether. Electrophoresis of aq. extract (0.05 M, pH 7 phosphate buffer, 50 V/cm., 20 min.) shows no free Thienamycin present. The aq. extract, adjusted to pH 6.8, contains 105 ODU at 302 nm. by uv analysis, which is 95% extinguished after treatment with hydroxylamine for one hour. NMR (.delta., D.sub.2 O) 1.56 (m, CHCF.sub.3), 2.84-3.60 (CH.sub.2), 3.72 (s, OCH.sub.3), 4.29 (s, OCH.sub.2), 4.98 (s HDO).

EXAMPLE 5

N-(p-Nitrobenzyloxycarbonylthienamycin Lithium Salt

To Thienamycin (220 mg. in 60 ml. water at 0.degree. C.), is added successively, 679 mg. NaHCO.sub.3, 60 ml. dioxane and then with stirring 1.1 equivalents p-nitrobenzylchloroformate over a period of 1.5 minutes. The mixture is allowed to react 10 minutes, and is then extracted three times with cold ethyl ether. Electrophoresis (0.05 M, pH 7, phosphate buffer, 50 V/cm., 20 minutes) shows no free Thienamycin present. The aq. extract is adjusted to pH 2.2 with 1 M H.sub.3 PO.sub.4 solution and extracted three times with EtOAc. The EtOAc extract is dried over MgSO.sub.4, filtered and reextracted 0.1 N LiOH, to pH 8.2. The final pH is adjusted to 7.0 with 1 M H.sub.3 PO.sub.4 and the sample lyophilized. The yield is 205 mg. (54%).

EXAMPLE 6

Preparation of N-(p-nitrobenzyloxycarbonyl)-Thienamycin p-t-butyl benzyl Ester

N-(p-nitrobenzyloxycarbonyl)Thienamycin Li salt, the product of Example 5 (205 mg) in 2 ml. hexamethylphosphoramide (HMPA) is treated for 2.5 hrs. with 0.1625 ml. p-t-butylbenzyl bromide. The starting material is insoluble in HMPA but goes into solution after 30 minutes.

The reaction mixture is diluted with ethyl acetate (EtOAc), washed successively with water, aqueous K.sub.2 HPO.sub.4, water, saturated aqueous NaCl, dried over MgSO.sub.4, filtered evaporated and subjected to preparative thin layer chromatography on silica gel; eluting with 1:2 CHCl.sub.3 :EtOAc. Yield 160 mg (58%), Rf 0.38, IR (.mu.film) 2.98 NH and OH 5.63, .beta.-lactam; 5.86 broad easter and urethane; NMR (.delta., CDCl.sub.3), 1.24 (s, CHCH.sub.3, t-butyl) 2.59-3.27 (m, CH.sub.2) 3.83-4.47 (m, CH .beta.-lactam), 5.15 (s OCH.sub.2 C.sub.6 H.sub.4 NO.sub.2), 5.22 (s OCH.sub.2 C.sub.6 H.sub.4 t-butyl (7.45 and 8.12 (AB quartet, J=8 Hz C.sub.6 H.sub.4 NO.sub.2)

EXAMPLE 7

Preparation of N-(p-Nitrobenzyloxycarbonyl)Thienamycin m-Phenoxybenzyl Ester

Following the procedure of Example 6, the title compound is prepared when an equivalent amount of m-phenoxybenzyl bromide is substituted for the p-t-butylbenzyl bromide of Example 6, Yield 11%, IR (.mu.film) 3.0 NH.sub.2, and OH 5.63 .beta.-lactam; 5.86 broad peak ester and urethane; nmr (.delta.CHCl.sub.3) 1.33 (d, CHCH.sub.3 J=6); 2.60-3.62 (m, CH.sub.2), 7.45 and 8.13 (AB quartet, J=8, C.sub.6 H.sub.4 NO.sub.2); 7.26 (s C.sub.6 H.sub.4 OC.sub.6 H.sub.5) M.S. m/e 589, 559, 547, 183.

EXAMPLE 8

Preparation of:

(1) N-(O-Formyl-D-mandeloyl Thienamycin p-t-butylbenzyl ester (2) N-(O-Formyl)-D-mandeloyl Thienamycin m-phenoxybenzyl ester (3) N-D-Mandeloyl Thienamycin p-t-butylbenzyl ester (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.

Following the procedure of Examples 6 and 7, title compounds 1., 3., 5., 7 and 2., 4., 6., 8 are prepared, respectively, when the appropriate N-acyl thienamycin starting material from Examples 1-4 replaces, in equivalent amount the N-(p-nitrobenzyloxycarbonyl) thienamycin starting material of Examples 6 and 7, respectively.

EXAMPLE 9

Preparation of N-(p-Methoxybenzyloxycarbonyl)thienamycin p-t-Butylbenzyl Ester

Step A: N-(p-methoxybenzyloxycarbonyl)thienamycin Sodium salt (I) and Lithium salt (II)

To Thienamycin (20 mg.) in 5 ml. water at 0.degree. C.: is added 105 mg. NaHCO.sub.3 (20 equivalents), 5 ml. dioxane, and then, dropwise with stirring over 1 min. ten equivalents of p-methoxybenzyl chloroformate. After 15 min. the pH is adjusted to 7.5 with 1 M H.sub.3 PO.sub.4 and the solution extracted 3X with ether. The aqueous portion is then adjusted to pH 2.2 at 0.degree. C. and extracted 3X with ethylacetate (EtOAc). The EtOAc is dried quickly with MgSO.sub.4, filtered and extracted with a few ml. water containing 6.3 mg. NaHCO.sub.3. The aqueous extract, lyophilized, contains 172 ODU at 303 nm by UV analysis in H.sub.2 O at pH 7.0, which is 95% extinguished after treatment with hydroxlamine for one hour. The yield is 16 mg. Electrophoresis (50 v/cm, 20 min., pH 7 aqueous phosphate, 0.05 M) shows one spot by bioautograph, 4 cm towards the anode. NMR (.delta., D.sub.2 O): 1.49 (d, J=6 Hz CH.sub.3 CH); 2.8-3.7 (m,CH.sub.2); 3.99 (s, OMe); 4.0-4.6 (m, .beta.-lactam CH); 4.92 (s, HDO); 5.20 (s, OCH.sub.2); 7.13 (d, J=8 Hz C.sub.6 H.sub.4).

The lithium salt II is made in the same way, but extracting the EtOAc solution with 0.1 N LiOH to pH 7.8 (instead of aqueous NaHCO.sub.3), and lyophilizing. The spectral and electrophoretic properties of II are the same as those of I.

Step B: N-(p-methoxybenzyloxycarbonyl)thienamycin-p-t-benzyl ester

The lithium salt of Step A (37 mg.) in 0.4 ml. hexamethylphosphoramide (HMPA), is treated for 2 1/5 hours with 0.033 ml. p-t-butylbenzyl bromide. The lithium salt is insoluble in HMPA but goes into solution after 15 minutes reaction time.

The reaction mixture is diluted with EtOAc, washed successively with water twice, aqueous K.sub.2 HPO.sub.4, water and brine, dried with MgSO.sub.4, filtered, evaporated and subjected to preparative layer chromatography on silica gel, eluting with 1:2 CHCl.sub.3 -EtOAc, affording 47 mg. pure II, Rf=0.3. IR (.mu., film): 3.0, NH; 5.63, .beta.-lactam; 5.87 broad, ester and urethan. NMR (.delta., CDCl.sub.3): 1.21 (s, Me and t-butyl); 2.6-3.6 (m, CH.sub.2); 3.72 (s, OMe); 3.8-4.4 (m, .beta.-lactam CH); ##STR33## 6.84 and 7.20 (AB quartet, C.sub.6 H.sub.4 OMe); 7.32 (S, C.sub.6 H.sub.4 -t-Bu). MS: 582, 538, 496.

EXAMPLE 10

Preparation of N-(p-methoxybenzyloxycarbonyl)thienamycin Benzhydryl Ester

To Thienamycin (23.5 mg.) in 5 ml. water is added successively 4 ml. dioxane, 62 mg. NaHCO.sub.3, and then, in portions at 0.degree. C. with stirring, 4 equivalents p-methoxybenzyl chloroformate over 4 minutes. After ten minutes total reaction time, the pH is adjusted to 7.0 with 1 N H.sub.3 PO.sub.4 and the mixture extracted three times with ether. Electrophoresis of the aqueous portion (0.05 M pH 7 aqueous phosphate buffer, 50 V/cm, 20 minutes) shows 50% conversion to N-(p-methoxybenzyloxycarbonyl)thienamycin.

The aqueous solution is brought to pH 2.2 with 1 M H.sub.3 PO.sub.4 at 0.degree. C. and extracted 3.times. with EtOAc. The EtOAc solution is treated with 50 mg. diphenyldiazomethane, evaporated and taken up in CH.sub.3 CN. More diphenyldiazomethane is added to a persistent purple color. After 0.5 hour the solution is evaporated and chromatographed on silica gel, eluting with 1:2 CHCl.sub.3 -EtOAc, affording 10 mg. pure title compound Rf 0.25. IR (.mu., film): 3.0, NH; 5.63, .beta.-lactam; 5.85, 5.89, ester and urethan. NMR .delta., CDCl.sub.3); 1.23 (s, OH); 1.30 (d, J=6 Hz, CH.sub.3 CH); 2.6-3.6 (m, CH.sub.2); 3.78 (s, OMe); 5.02 (s OCH.sub.2); 3.8-4.4 (m, .beta.-lactam CH); 6.9 and 7.35 (AB quartet, J=9 Hz, C.sub.6 H.sub.4), 7.3s CHPh.sub.2.

EXAMPLE 11

Preparation of N-(o-Nitrobenzyloxycarbonyl)Thienamycin Benzyl Ester

Step A: N-(o-Nitrobenzyloxycarbonyl)thienamycin Sodium Salt

To Thienamycin (43 mg.) at 0.degree. C. is added 10 ml. 1:1 tetrahydrofuran (THF:Water). The mixture is rapidly stirred while 102 mg. NaHCO.sub.3 (10 equivalents) is added, and then, dropwise with stirring over 2 minutes, four equivalents of o-nitrobenzylchloroformate is added. After 30 minutes, the pH is adjusted to 7 with aqueous 25% H.sub.3 PO.sub.4 and the solution extracted three times with ether. The aqueous layer is evaporated at 25.degree. C., in vacuo and is then adjusted to pH 2.2 at 0.degree. C. Solid NaCl is added, and the cold acidic solution is extracted 3.times. with cold EtOAc. The EtOAc extracts are combined and quickly back-washed with cold brine; dried with MgSO.sub.4, filtered and back extracted with 10 ml. of water containing 1.75 equivalents of solid NaHCO.sub.3. The extract is lyophilized in vacuo at 25.degree. C. to provide the title compound.

Step B: N-(o-Nitrobenzyloxycarbonyl)Thienamycin Benzyl Ester

The product of Step A in 7.5 ml. EtOAc (from the pH 2.2 extraction) is treated with an excess of phenyldiazomethane (4 ml. of solution comprising 20 mg./ml. ether) at 4.degree. C. for 2.3 hours. The mixture is concentrated to wet residue at 20.degree. C. under reduced pressure. The desired compound is isolated by thin layer chromatography, EtOAc; ether (9:1) to afford 17.5 mg. of N-(o-Nitrobenzyloxycarbonyl)thienamycin benzyl ester.

EXAMPLE 12

Preparation of N-(o-Nitrobenzyloxycarbonyl)Thienamycin p-methoxy Benzyl Ester

To N-(o-Nitrobenzyloxycarbonyl)thienamycin (70 mg.) in 8 ml. of EtOAc is added 4 ml. of p-methoxyphenyldiazomethane (9 mg./ml. acetonitrile) at 4.degree. C. The mixture is stirred for 1.5 hours at 4.degree. C. and is then concentrated to a wet paste under reduced pressure at 20.degree. C. The title compound (42 mg.) is isolated by thin layer chromatography silica gel, eluting with EtOA:ether (9:1).

EXAMPLE 13

Preparation of N-(o-nitrobenzyloxycarbonyl)thienamycin p-Bromo-phenacyl Ester ##STR34##

To Th (TMS).sub.3, I, (24 mg.), which is prepared according to Example 17, in 0.8 ml. dry THF is added 23 mg. o-nitrocarbobenzyloxy chloride, followed by 0.015 ml. of triethylamine. After vibro-mixing for 30 minutes at 25.degree. C., the mixture is concentrated to a pasty residue in a stream of dry N.sub.2, and is washed 3.times. with petroleum ether. The residue is suspended in 1 ml. of dry THF and p-bromophenacylbromide (14 mg.) is added followed by 0.03 ml. triethylamine. After vibro-mixing for 30 minutes at 25.degree. C., the mixture is evaporated to dryness in vacuo at 20.degree. C. The residue is dissolved in EtOAc (2 ml) and shaken with 0.3 ml. of pH 4 buffer for 5 minutes. The organic layer is dried over MgSO.sub.4, filtered, evaporated to a pasty residue and the desired product is isolated (44 mg) by preparative thin layer chromatography on silica gel, eluting with EtoAc:CHCl.sub.3 (7:3).

EXAMPLE 14

Preparation of N-(Trichloroethoxycarbonyl)thienamycin Benzyl Ester

Step A: N-(Trichloroethoxycarbonyl)Lithium Salt

To Thienamycin (40 mg.) in 18 ml. 1:1 THF-H.sub.2 O at 0.degree. C. is added while stirring 255 mg. (15.2 equivalents) NaHCO.sub.3, and then, dropwise with stirring over 2 min., 1.8 equivalents of trichloroethylchloroformate dissolved in 0.6 ml. THF. After 6 minutes the pH is adjusted to 7.2 with aqueous 25% H.sub.3 PO.sub.4 and the solution extracted with ether. The aqueous portion after removing any entrained ether in vacuo is then brought to pH 2.5 at 0.degree. C. and extracted with cold EtOAc. The ethyl acetate extracts are combined, quickly backwashed with cold brine, dried with anhydrous MgSO.sub.4, filtered and back extracted with 0.01 m LiOH to pH 6.8. The aqueous extract is freed from any EtOAc in vacuo and lyophilized. The residual product contains 936 ODu (39.7%) by uv analysis at 302 nm which is 90% extinguished after treatment with hydroxylamine for one hour in 0.05 M phosphate buffer (pH 7). The yield is 32 mg. Electrophoresis (50 volts/cm. 20 min., pH 7 aq. phosphate 0.05 M) exhibits one zone by bioautograph (MB 108, staph. aureus), 2.4 cm toward the anode. Liquid chromatography C.sub.18 Bondapak (Waters Assoc.) in aqueous 10% THF exhibits one main peak free of any unreacted Thienamycin.

Step B: N-(Trichloroethoxycarbonyl)thienamycin Benzyl Ester

The compound of Step A (32 mg.) in 2 ml. dry distilled DMF containing 7% HMPA (dry, pH 6.3), is treated with 0.015 ml. benzyl bromide for 2 hours at 15.degree. C. (allowing the contents to warm up to 25.degree. C. during the course of the reaction). The reaction mixture is diluted with EtOAc, washed successively with cold H.sub.2 O, 1% aqueous NaHCO.sub.3, water and cold saturated aqueous NaCl, dried with MgSO.sub.4, filtered, evaporated and subjected to preparative thin layer chromatography on silica gel, eluting with 1% CH.sub.3 CN in EtOAc to afford 10 mg. of the title compound, Rf=0.63; IR (.mu. CHCl.sub.3) 5.63, .beta.-lactam; 5.78 and 5.88 broad ester and urethane. NMR (.delta. CDCl.sub.3) 1.35 (d, Me); 2.8-3.7 (m CH.sub.2); 3.51 and 4.27 (dd, J=6 Hz, .beta.-lactam CH); 4.79 (s, OCH.sub.2 CCl.sub.3) 5.42 s(OCH.sub.2 C.sub.6 H.sub.5); and 7.41 (m, C.sub.6 H.sub.5).

EXAMPLE 15

Preparation of N-Bromoacetyl thienamycin Methyl and Benzyl Esters

Step A: N-bromoacetyl thienamycin

To a cooled solution of thienamycin (28.8 mg.) and sodium bicarbonate (0.3 g.) in 10 ml. of water and 8 ml. of dioxane is added with stirring a solution of 0.25 g. of bromoacetic anhydride in 2 ml. dioxane over a period of 20 minutes. The pH is maintained at 8.0. The mixture is stirred for an additional 5 minutes then layered with 10 ml. of ether and the pH adjusted to 7 by the additional of 8% phosphoric acid. The ethereal layer is separated and the aqueous layer is extracted twice again with ether. The aqueous layer is evaporated under reduced pressure to 0.5 ml., diluted to 2 ml. with water and put on 50 ml. of XAD-2 resin.

The column is eluted with water. The first 80 ml. is discarded, then the next 100 ml. is collected. The solvent is changed to 10% THF and an additional 100 ml. collected. The combined eluates are adjusted to pH 7, evaporated to 5 ml. under reduced pressure, then freeze-dried to give the sodium salt of N-bromoacetyl thienamycin in 60% yield. UV .lambda..sub.max 302 m.mu..

Step B: N-Bromoacetyl thienamycin Methyl and Benzyl Esters

An aqueous solution of the sodium salt is layered with ethyl acetate at 0.degree. C. and adjusted to pH 2. The ethyl acetate phase is separated and the aqueous phase is extracted with ethyl acetate. The combined ethyl acetate solutions are dried over MgSO.sub.4 and then treated with a solution of diazomethane. The solvents are evaporated and the residue chromatographed on a silica gel plate. R.sub.f 0.11 in 2:1 ethyl acetate-chloroform. m.p. 118.degree.-120.degree. C. Mass spectrum shows M.sup.+ at m/e 406 and significant fragments at m/e 362, 320, 183 and 164.

The corresponding benzyl ester is prepared in a similar way from phenyldiazonemethane. m.p. 142.degree.-3.degree. C.

Ir: 5.65.mu., 5.89.mu. and 6.1.mu.. Mass spec. M.sup.+ m/e 482 also m/e 438, 396, 316, 259 and 164.

EXAMPLE 16

Preparation of N-(Guanylthioacetamido)thienamycin Methyl and Benzyl Esters ##STR35##

To a solution of 20 mg. of N-bromoacetyl thienamycin methyl ester in 5 ml. of aqueous 80% dioxane, 10 mg. of thiourea is added. The solution is kept at 4.degree. C. for five days and is then chromatographed on 80 ml. of XAD-2 resin. Elution with 400 ml. of water removes thiourea. Elution with 120 ml. of aqueous 10% tetrahydrofuran gives a solution containing the N-guanylthioacetacmidothienamycin methyl ester. High pressure liquid chromatographic (HPLC) analysis on Bondapak C.sub.18 porasil with aqueous 10% THF solvent shows a major peak of 90% purity. The solution is evaporated to 15 ml. (pH 5.5) and freeze-dried. The corresponding N-guanylthioacetamido thienamycin benzyl ester is prepared by following the same procedure except that the benzyl ester of Example 15 is substituted for the methyl ester.

EXAMPLE 17 ##STR36##

Preparation of Silylated-Th

Thienamycin (80.0 mg.) is suspended in 40 ml. tetrahydrofuran (THF) under a N.sub.2 atmosphere and is concentrated to 10 ml.; hexamethyldisilazane (1.0 ml.) and trimethylchlorosilane (300 .mu.l) is added. The mixture is reacted for 20 mins. at 25.degree. C. with vigorous stirring. The suspension is then centrifuged to remove ammonium chloride. The supernatant is evaporated to an oil under a nitrogen stream for future reaction.

EXAMPLE 18

Preparation of N-(Bromo-t-Butoxycarbonyl)thienamycin p-Bromophenacyl Ester

Step A: Preparation of N-Bromo-t-Butoxycarbonyl-O-TMS-Tienamycin-TMS Ester

Th(TMS).sub.3 (16 mg.) is dissolved in 0.4 ml. of dry tetrahydrofuran to which is added 20 .mu.l (28 mg., 0.13 mmol) of bromo-t-butylchloroformate (b.p. 35.degree./0.9 mm) and 8 .mu.l (5.67 mg., 0.057 mmole) of triethylamine (redistilled from BaO). The mixture is shaken at 25.degree. C. for 20 min. Evaporation of solvent and excess reagents gives crude desired product. UV .mu..sub.max.sup.CH.sbsp.3.sup.CO.sbsp.2.sup.CH.sbsp.2.sup.CH.sbsp.3 320 nm (E9,000).

Step B: Preparation of N-(Bromo-t-Butoxycarbonyl) thienamycin

The product of Step A (3 mg.) is dissolved in 0.5 ml. of ph 7 phosphate buffer and 0.1 ml. of tetrahydrofuran and the solution left at 25.degree. C., for 20 minutes. The solution is then passed down a column (5 ml.) of Dowex 50.times.8 (Na.sup.+ form) and the eluate fractions monitored by U.V. The correct fractions are combined and freeze dried to yield the desired product. UV .lambda..sub.max.sup.buffer 304 nm (.epsilon.=9,300); electrophoresis at 50 V/Cm. 20 min in pH 7.0 buffer shows a single bioactive zone which moves 31.5 mm towards the anode.

Step c: Preparation of N-(Bromo-t-Butoxycarbonyl) thienamycin p-Bromophenacyl Ester

The product of Step B (13 mg. 0.022 mmole) is dissolved in 0.4 ml. of tetrahydrofuran. To this solution is added p-bromophenacyl bromide (9.6 mg., 0.035 mmol) and 20 .mu.l (14.4 mg., 0.14 mmole) of triethylamine. The mixture is shaken at 25.degree. C. for 30 min. and then evaporated to dryness. Ten ml. of ether is added to the residue and the mixture treated with 0.2 ml. of 0.1 M pH 7.0 phosphate buffer.

The organic layer is separated, dried over sodium sulfate, concentrated to 0.5 ml. and applied to two 20.times.20 cm., 250.mu. silica gel GF tlc plates which are developed with 20% ethyl acetate in chloroform. (R.sub.f =0.65) the desired product (6.7 mg.) is isolated in 42% yield.

EXAMPLE 18a

Preparation of N-Bromo-t-butoxycarbonyl Thienamycin Sodium Salt

Method A:

Thienamycin (190 mg) dissolved in 15 ml 0.1 M pH 7.0 phosphate buffer and 15 ml dioxane is kept at 0.degree. C. The solution is adjusted and maintained between pH 8.5-9.0 with 1 N NaOH while 480 mg of bromo-t-butyl chloroformate is added to the solution during a period of 5 minutes. The mixture is stirred for 30 min., then is neutralized to pH 7.0 with 1 N HCl and extracted with ether. The aqueous layer is separated, concentrated to 10 ml and chromatographed on a Dowex-50.times.8 (Na form) column (1.5".times.10") which is eluted with H.sub.2 O to give 113 mg of the desired product. Lyophilization of the solution gives solid product.

Method B:

Thienamycin (95 mg) in 10 ml 0.1 M phosphate buffer and 10 ml dioxane is kept at 0.degree. C. The solution is adjusted and maintained between pH 8.5-9.0 while 240 mg of bromo-t-butyl chloroformate is added. The mixture is stirred for 30 minutes, then is acdified to pH 2.0 with H.sub.3 PO.sub.4. The acidified solution is extracted with 2.times.25 ml ethylacetate. The organic layer is separated and back extracted with 10 ml NaHCO.sub.3 solution which contains 30 mg of NaHCO.sub.3. The aqueous layer containing 30 mg of the desired product is lyophilized to give solid product. Nmr (60 MHz, D.sub.2 O): .delta.1.26(d), 1.60(s), 2.65-3.50(m), 3.70(s), and 3.90-4.20(m). UV .lambda..sub.max.sup.D.sbsp.2.sup.O 303 nm.

EXAMPLE 18b

Preparation of N-Bromo-t-butoxycarbonyl Thienamycin p-Nitrobenzyl Ester

The lyophilized N-bromo-t-butoxycarbonylthienamycin sodium salt (100 mg) is stirred at 25.degree. C., with p-nitrobenzyl bromide (300 mg) in 2 ml hexamethylphosphoramide for 1 hour. The mixture is diluted with 10 ml ethylacetate then is washed thoroughly with water. The organic layer is separated, dried over Na.sub.2 SO.sub.4 and chromatographed on two 250.mu. silica gel GF TLC plates using ethylacetate as solvent (R.sub.f 0.45) to give 50 mg of the desired product. IR (CDCl.sub.3): 1777 (.beta.-lactam) and 1711 cm.sup.-1 (ester): UV .lambda..sub.max.sup.EtOH 270 nm and 322 nm; NMR (CDCl.sub.3, 60 MHz): .delta.1.38(d), 1.58(s), 2.60-3.80(m), 3.78(s), 3.90-4.20(m), 5.30(s), 7.55(d) and 8.30 ppm (d).

EXAMPLE 18c

Preparation of N-(Dimethoxyphosphinothioyl)-Thienamycin Pivaloxymethyl Ester ##STR37##

To 32 mg of N-(dimethoxyphosphinothioyl)thienamycin sodium salt in 2 ml of dry hexamethylphosphoramide (HMPA) is added under N.sub.2 2.2 eq (25 mg) of chloroethyl pivalate in 0.3 ml HMPA. (Added in 3 portions over 15-20 minutes). The reaction mixture is stirred at 25.degree. C., for 2 hours, diluted with EtOAc (20 ml), washed 5.times.6 ml H.sub.2 O, 1.times.4 ml KH.sub.2 PO.sub.4, 2.times.6 ml H.sub.2 O and 1.times.7 ml cold brine and dried over MgSO.sub.4. After filtering the EtOAc extract is concentrated in vacuo to give 70 mg of an oil Preparative thin layer chromatography (1% MeOH-EtOAc) gives 14.5 mg (37%) of N-(dimethoxyphosphinothioyl)-thienamycin pivaloxymethyl ester. R.sub.f 0.67 UV maximum at 325 nm; IR(CHCl.sub.3 C.dbd.O 1785 (5.60) and P-OCH.sub.3 asymm. stretch at 970 cm.sup.-1 (10.3.mu.) NMR(CDCl.sub.3 100 MHz) exhibits a strong singlet 1.15 for D(CH.sub.3).sub.3 (9H) a doublet at 8.71 ppm for P-OCH.sub.3 (J=13H.sub.z).

EXAMPLE 18d

Preparation of N-p-Nitrobenzyloxycarbonyl Thienamycin Benzyl Ester

A solution of thienamycin (115 mg) in 2 ml water and 2 ml dioxane is cooled in an ice bath and titrated to pH 5 with 1.0 N H.sub.2 SO.sub.4. A solution of phenyldiazomethane (0.8 mmole) in 2 ml dioxane is added with rapid stirring during a period of 2 minutes to the thienamycin solution. The pH is kept at 5-5.5 with an automatic titrator. A solution of 100 ml NaHCO.sub.3 in 2 ml of water is added and the pH is adjusted to 8.2 with 1.0 N NaOH. A solution of 300 mg. p-nitrocarbobenzyloxy chloride in 2 ml dioxane is added during 2 minutes, while the pH is maintained at 8.2. After an additional 10 minutes, the reaction mixture is poured into 50 ml water and 50 ml ethylacetate. The ethylacetate layer is separated and washed with brine and dried over magnesium sulfate and evaporated. The residue is chromatographed on 3" .times.8" 1000.mu. silica gel plates which are developed with 1:1 ethylacetate/chloroform. The bands at 1.5-3.0 cm are eluted with ethylacetate. Evaporation of the ethylacetate yields crystalline N-p-nitrobenzyloxycarbonyl thienamycin benzyl ester. Yield 30 mg.

m.p. 165.degree.-170.degree. C.; TLC 1:1 ethyl/chloroform R.sub.f 0.16; U.V. .lambda..sub.max.sup.ethylacetate 270 m.mu. E%=174, .lambda..sub.min =294 m.mu. E%=146, .lambda..sub.max 319 m.mu. E%=174; I.R. 5.65.mu. (lactam).

EXAMPLE 19

Preparation of N-Azidoacetyl thienamycin Sodium salt (I) Lithium Salt (II)

I. Na Salt

Thienamycin (48 mg., 0.18 mmole) is dissolved in 10 ml. cold water and is kept at 0.degree. C. To the solution is added sodium bicarbonate (147 mg., 17.6 mmole) and dioxane (10 ml.). Azidoacetyl chloride (60 mg., 0.50 mmole) is added to the solution during a period of 2 min. The reaction mixture is stirred for 15 min. neutralized to pH 7.0 with 30% phosphoric acid and then transfered into a separatory funnel. The solution is extracted with 2.times.50 ml. ether. The aqueous layer is concentrated to 5 ml. and then is charged to a Dowex AG-50.times.8 (sodium form) ion exchange column. The desired fractions, monitored by U.V., are combined and lyophilized to give 21 mg. of the product. Electrophoresis (50 V/cm, 20 min., in pH 7 phosphate buffer) of the product shows a single bio-active band which moves 40 mm towards anode in pH 7.0 phosphate buffer. Uv .lambda..sub.max.sup. H.sbsp.2.sup.O 300 nm; Pmr (100 MHz, D.sub.2 O) 1.26 (d, CH.sub.3 CH), 2.92-3.43 (m, 3CH.sub.2 and C.sub.6 -H), 4.01 (s, CH.sub.2 N3) and 4.20 ppm (m, C.sub.5 -H and C.sub.7 -H).

II. Li Salt

Thienamycin (76.2 mg., 0.28 mmol) is dissolved in 10 ml. of cold water and is kept at 0.degree. C. To the solution is added 0.6 ml. of 1.0 N lithium hydroxide solution and 10 ml. dioxane. After stirring for 1 min., azidoacetyl chloride (33.6 mg., 0.28 mmol) is added during a period of 2 min. The reaction mixture is stirred for an additional one minute then is neutralized to pH 7.0 with 30% phosphoric acid. After extraction with ether, the aqueous solution is concentrated to 5 ml. and is charged to the Dowex AG-50wx8 (lithium form) ion exchange column. The desired fractions, monitored by U.V., are combined and lyophilized to give 38 mg. of the product. Uv .lambda..sub.max.sup.H.sbsp.2.sup.O 300 nm.

EXAMPLE 20 ##STR38##

Preparation of N-Glycyl thienamycin

Platinum oxide (60 mg.) and 2 ml. water are placed in a hydrogenation flask and stirred under 1 atm. hydrogen at 25.degree. C. for 20 minutes. To the flask is added N-azidoacetyl thienamycin sodium salt (6.0 mg., 0.02 mmole) in 4 ml. water. The reaction mixture is stirred at 25.degree. C. under 1 atm. hydrogen for 30 minutes. The resulted mixture (pH 8.7) is adjusted to pH 7.0 with 30% phosphoric acid then is filtered from the catalyst. The aqueous solution is concentrated to 2 ml. and then is charged to the Dowex AG-50wx8 (sodium form) ion exchange column. The desired fractions monitored by U.V. are combined and lyophilized to give 3.8 mg. of the product. Electrophoresis (50 V/cm, 20 min.) of the product in pH 7.0 phosphate buffer shows one single bio-active band at the origin. Uv.sub.max.sup.H.sbsp.2.sup.O 300 nm; Pmr (100 MHz, D.sub.2 O): 1.27 (d, CH.sub.3 CH), 2.96-3.37 (m, 3CH.sub.2 and C.sub.6 -H), 3.70 (s COCH.sub.2 NH.sub.2), and 4.20 ppm (m, C.sub.5 -H and C.sub.7 -H).

EXAMPLE 21

Preparation of N-Azidoacetyl thienamycin Benzyl Ester ##STR39##

N-Azidoacetyl thienamycin lithium salt (3.0 mg.) is stirred with hexamethylphosphoramide (HMPA) (1.0 ml.) and benzyl bromide (30 mg., 0.21 mmole) for 30 minutes at 25.degree. C. The reaction mixture is then diluted with ethyl acetate (5 ml.) and washed thoroughly with water. The organic layer is separated, and dried over sodium sulfate. The product (2.0 mg.) is isolated by silica gel TLC (R.sub.f =0.18 in ethyl acetate). IR (CHCl.sub.3): 2125 (N.sub.3), 1777 (.beta.-lactam) and 1687 cm.sup.-1 (ester and amide); Pmr (CDCl.sub.3, 100 MHz); 1.34 (d, CH.sub.3 CH), 2.80-3.60 (m, 3CH.sub.2 and C.sub.6 -H), 3.97 (s, CH.sub.2 N.sub.3), 4.21 (m, C.sub.5 -H and C.sub.7 -H), 5.19 and 5.35 (d, CH.sub.2 C.sub.6 H.sub.5), 6.63 (m, NH) and 7.32 (m, C.sub.6 H.sub.5). N-Azidoacetyl thienamycin sodium salt (30 mg., 0.08 mmole) is stirred with HMPA (3 ml.) and benzyl bromide (120 mg., 0.07 mmole) at 25.degree. C., for 30 min. The product (30 mg.) is isolated by the same procedure described above.

EXAMPLE 22

Preparation of Preparation of N-Azidoacetyl thienamycin 3-Methyl-2-butene-1-yl Ester

N-Azidoacetyl thienamycin sodium salt (11.0 mg., 0.029 mmole) is stirred with HMPA (1 ml.) and 1-bromo-3-methyl-2-butene (39 mg., 0.26 mmole) at 25.degree. C. for 30 min. The mixture is then diluted with 10 ml. ethyl acetate and washed thoroughly with water. The desired product (10 mg.) is isolated by silica gel TLC (R.sub.f =0.18 in ethyl acetate). Ir (CHCl.sub.3): 2121 (N.sub.3), 1777 (.beta.-lactam) and 1685 cm.sup.-1 (ester and amide); Pmr (CDCL.sub.3, 100 MHz): 1.34 (d, CH.sub.3 CH), 1.73 (s, .dbd.C(CH.sub.3).sub.2, 2.80-3.80 (m, 3CH.sub.2 and C.sub.6 -H), 3.98 (s, CH.sub.2 N.sub.3), 4.20 (m, C.sub.5 -H and C.sub.7 -H), 4.72 (d, CH.sub.2 CH.dbd.), 5.40 (t, CH2CH.dbd.), and 6.70 (broad, NH); Mass Spect (E. I.): m/e 423 (molecular ion), 405 (M.sup.+ -H.sub.2), 395 (M.sup.+ -N.sub.2), and 337 (M.sup.+ -86).

EXAMPLE 23

Preparation of N-Glycyl thienamycin 3-Methyl-2-butene 1-yl Ester ##STR40##

N-Azidoacetyl thienamycin 3-methyl-2-butene-1-yl ester (10 mg.) is dissolved in ethyl acetate (1 ml.). This solution is aded to the hydrogenation flask containing 10 mg. of palladium (from palladium oxide) and 0.5 ml. 50% methanol in ethyl acetate. The mixture is stirred under 1 atm hydrogen at 25.degree. C. for 1 hour, then the mixture is filtered from the catalyst. The desired product is isolated by TLC (R.sub.f =0.16, in 20% methanol/chloroform). Electrophoresis of the product at pH 7.0 buffer (50 V/cm, 20 min.) shows one bio-active band which moves 34 mm towards the cathode Uv .lambda..sub.max.sup.ethanol 315 nm, IR (CHCl.sub.3): 3300 (NH.sub.2), 1777 (.beta.-lactam), and 1672 cm.sup.-1 (ester and amide).

EXAMPLE 24

Preparation of N-Azidoacetyl thienamycin p-t-Butylbenzyl Ester

N-Azidoacetyl thienamycin sodium salt (136 mg., 0.36 mmole) stirred with HMPA (5 ml.) and p-t-butylbenzyl bromide (180 mg., 0.79 mmole) at 25.degree. C. for 30 min. The solution is diluted with 10 ml. ethyl acetate and washed thoroughly with water. The organic layer is separated and dried over sodium sulfate. The desired product (80 mg.) is isolated by silica gel TLC (R.sub.f =0.18 in ethyl acetate). IR (CHCl.sub.3): 2121(N.sub.3) 1777 (.beta.-lactam) and 1684 cm.sup.-1 (ester and amide); Pmr (CDCl.sub.3, 60 MHz): 1.32 (s, t-Bu), 1.34 (d, CH.sub.3 CH), 2.60-3.75 (m, 3CH.sub.2 and C.sub.6 -H), 3.90 (s, C.sub.2 N.sub.3), 4.30 (m, C.sub.5 -H and C.sub.7 -H), ##STR41## and 7.33 ppm (s, aromatic protons).

EXAMPLE 25

Preparation of N-Glycyl thienamycin p-t-Butylbenzyl Ester

N-Azidoacetyl thienamycin p-t-butylbenzyl ester (10 mg.) is dissolved in 0.5 ml. ethyl acetate. The solution is added to the hydrogenation flask containing 50 mg. of palladium (from palladium oxide) and 0.5 ml. ethyl acetate. The mixture is stirred under 1 atm hydrogen at 25.degree. C. for 10 minutes. TLC indicates (as in Example 24) that all the starting material is consumed. The reaction mixture is filtered from the catalyst and evaporated to dryness to give the crude product. IR (CHCl.sub.3): 1776 (.beta.-lactam) and 1675 (ester and amide); Uv .sub.max.sup.ethanol 320 nm and 275 nm; Pmr (CHCl.sub.3 60 MHz); 1.32 (s, t-Bu), 1.34 (d, CH.sub.3 CH), ##STR42## and 7.33 ppm (s, aromatic protons).

EXAMPLE 26

Preparation of N-Azidoacetyl thienamycin Pivaloxymethyl Ester ##STR43##

N-Azidoacetyl thienamycin sodium salt (11.0 mg., 0.04 mmole) is stirred with HMPA (1 ml.) and chloromethyl pivalate (36 mg., 0.24 mmole) at 25.degree. C. for 30 min. The mixture is diluted with ethyl acetate and washed with water. The desired product is isolated by silica gel TLC (R.sub.f =0.18 in ethyl acetate)IR (CHCl.sub.3): 2121 (N3), 1777 cm.sup.-1 (.beta.-lactam); Pmr (CDCl.sub.3, 60 MHz); 1.22 (s,t-Bu), 1.32 (d, CH.sub.3 CH), 3.98 (s, CH.sub.2 N.sub.3), and 5.83 ppm (dd, CO.sub.2 CH.sub.2 O-).

EXAMPLE 27

Preparation of N-Azidoacetyl thienamycin 2-Methyl-2-propene-1-yl Ester ##STR44##

N-Azidoacetyl thienamycin lithium salt (20 mg., 0.055 mmole) is stirred with HMPA (1 ml) and 3-chloro-2-methylpropene (27 mg. 0.30 mmole) for 30 min. at 25.degree.. The mixture is diluted with ethyl acetate and washed with water. The desired product is isolated by silica gel TLC (R.sub.f =0.18 in ethyl acetate). IR (CHCl.sub.3): 2121 (N.sub.3), 1777 (.beta.-lactam), and 1684 cm.sup.-1 (ester and amide).

EXAMPLE 28

Preparation of N-Glycyl thienamycin Esters (I-IV)

______________________________________ ##STR45## R______________________________________ ##STR46## II ##STR47## III ##STR48## IV ##STR49##______________________________________

Compounds I-IV are prepared respectively, when the corresponding starting material (Examples 22, 24, 26 and 27, respectively) are substituted in equivalent amount in the reduction process of Example 23, for the N-azidoacetyl thienamycin 3-methyl-2-butene-1-yl ester of Example 23.

EXAMPLE 29

Preparation of N-Benzyloxycarbonyl thienamycin and N-Benzyloxycarbonyl Benzylcarbonic Acid Anhdride ##STR50##

A solution of 16.6 mg. of Thienamycin in 4 ml. of 0.05 M pH 7 phosphate buffer and 2 ml of dioxane in a 3-necked flask fitted with a stirrer, thermometer, pH electrode and the delivery tip of an automatic titrator is cooled to -8.degree. C. in a methanol-ice bath. The pH is brought to 8.2 by the addition of 0.2 N sodium hydroxide in 50% aqueous dioxane and a solution of 0.015 ml of carbobenzyloxy chloride in 2 ml of chloroform is added. The mixture is stirred at -6.degree. C., pH 8.2, for ten minutes, then layered with ether and the pH adjusted to 7 by the addition of N hydrochloric acid. The layers are separated by centrifugation and the aqueous phase is extracted twice again with ether. The aqueous phase is layered with ethyl acetate and acidified to pH 2. The ethyl acetate is separated and the aqueous layer is extracted again with ethyl acetate. The combined ethyl acetate layer is washed with saturated sodium chloride solution, dried over magnesium sulfate and filtered. The filtrate is stirred with water and the pH brought to 7 by the addition of dilute sodium bicarbonate solution. The aqueous phase is separated and freeze-dried giving the sodium salt of N-benzyloxycarbonyl Th. Weight 10 mg (46%). The UV spectrum, .lambda..sub.max 303 m.mu., E% 147 (E 6,290) indicates about 80% purity. Electrophoresis at 50 V/cm. for 20 minutes at pH 7 followed by bioautograph on S. aureus gives a zone of inhibition at +2.5 cm.

The ethereal extracts of the reaction mixture contain the desired product N-benzyloxycarbonyl thienamycin benzyl carbonic acid anhydride. UV .lambda..sub.max 335 m.mu..

EXAMPLE 30

Preparation of N-Benzyloxycarbonyl thienamycin Benzyl Ester

The N-Benzyloxycarbonyl thienamycin [Example 29], in EtOAc is carried through the procedure of Example 29, except that an equivalent amount of phenyldiazomethane is added to the dried EtOAc solution from the pH 2 extraction and the solution left at 4.degree. for 2 hours. Evaporation to dryness yields crude N-benzyloxycarbonylthienamycin benzyl ester which is isolated by thin layer chromatography R.sub.f 0.24 3:1 ethyl acetate chloroform. It crystallizes from ether. IR 5.63.mu. (lactam carbonyl); shoulder 5.8.mu. (ester); 5.88.mu. (urethane carbonyl). UV, dioxane, .lambda..sub.max 318 m.mu., E% (.epsilon.=10,900) m/e M.sup.+ 496.

EXAMPLE 31

Preparation of N-Carbomethoxy thienamycin p-pivaloyloxybenzyl Ester

Step A: N-Carbomethoxy thienamycin

Thienamycin (59 mg., 148 .mu.mol) is dissolved in 0.05 M pH 7 phosphate buffer (14 ml.) 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 .mu.l, 600.mu. mol) in p-dioxane (580 .mu.l) is added at once to give a homogeneous solution. Subsequently, the pH is maintained at 8.2 using the automatic burette. After 10 min., the solution is adjusted to pH 7 using dilute phosphoric acid solution and washed three times with an equal volume of diethyl ether. The aqueous solution is then concentrated to 4.5 ml. and chromatographed on an XAD-2 resin column. The product is eluted (after water elution) with an aqueous 5% tetrahydrofuran solution and is freeze-dried to give 28.9 mg. of product. UV (pH 7 phosphate buffer 0.1 N). .lambda..sub.max 303 nm (e 6,450) Electrophoresis (20 min., 0.1 N pH7 phosphate buffer, 50 v/cm) mobility 3.5 cm to cathode.

Step B: N-Carbomethoxy thienamycin p-pivaloyloxybenzyl Ester

Following the procedure of Example 24 and substituting in equivalent amounts: N-carbomethoxy thienamycin (as its sodium salt) and p-pivaloyloxybenzyl bromide for the N-aziodoacetyl thienamycin sodium salt and p-t-butylbenzyl bromide, respectively, the title is obtained.

EXAMPLE 32

Preparation of N-Benzenesulfonyl thienamycin 2-Methyl)-2-propen-1-yl Ester ##STR51##

Step A: N-Benzenesulfonyl thienamycin

Thienamycin (52 mg. 148 .mu.mol) is dissolved in pH 7 0.1 N phosphate buffer (25 ml.) and magnetically stirred in an ice bath. The pH is adjusted to 8.2 with 2.5 N NaOH using an automatic dispensing burette and benzenesulfonyl chloride (227 .mu.l, 226 .mu.mol) in 500 ml. p-dioxane added at once. The pH is maintained at 8.2 (using the automatic burette) for 30 min. and then adjusted to pH 7.0 with 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 which elutes the product. The 10% aqueous tetrahydrofuran eluate is concentrated to 1/3 volume and freeze-dried to give 28 mg. Electrophoretic mobility of the product (50 v/cm, 20 min., pH 7 0.05 N phosphate buffer) is 3.5 cm towards the cathode. .lambda..sub.max 303 (3,650) in pH 7 0.1 N phosphate buffer).

Step B: N-Benzenesulfonyl thienamycin 2-Methyl-2-propen-1-yl Ester

Following the procedure of Example 24 and substituting in equivalent amounts N-Benzenesulfonyl thienamycin (as its sodium salt) and 2-methyl-2-propen-1-yl chloride for the N-azidoacetyl thienamycin sodium salt and p-t-butylbenzyl bromide respectively, the title compound is obtained.

EXAMPLE 33

Preparation of O-Methyl-Thienamycin ##STR52##

Step A

O-Methyl-N-(p-Nntrobenzyloxycarbonyl)-Thienamycin-(p-Nitrobenzyl)-Ester

To a solution of 135 mg. of N-p-nitrobenzyloxycarbonyl-Thienamycin-p-nitrobenzyl ester in 50 ml. of methylene chloride at 0.degree. C. is added with vigorous stirring 0.5 ml of 0.006 M fluoboric acid in ether-methylene chloride (3:1), immediately followed by 10 ml. of a cooled solution of 0.6 M diazomethane in methylene chloride. The diazomethane is decolorized in one minute. The solution is extracted with 10 ml. of 0.1 N pH 7 phosphate buffer, dried and evaporated to a small volume. The solution is applied to two 8".times.8" 1000.mu. silica gel plates which are developed with 3:1 ethylacetate-chloroform. The band at 3-4.5 cm yields 12 mg. of recovered starting material. The band at 6-8 cm yields 20 mg. of crystalline O-methyl-N-(p-nitrobenzyloxycarbonyl)-Thienamycin-(p-nitrobenzyl)ester. m.p. 172.degree.-174.degree. C. MS m/e 600 (M.sup.+), 568, 542, 500, 447, 389, 304 and 59.

Step B

O-Methyl-Thienamycin

A solution of 20 mg. of O-methyl-N-p-nitrobenzyloxycarbonyl-Thienamycin-p-nitrobenzyl ester in 2 ml. of tetrohydrofuran and 1 ml. of ethanol is hydrogenated at 50 psig, 23.degree. C. in the presence of 20 mg. of platinum oxide for 21/2 hours. The catalyst is filtered and 1 ml. of 0.1 N pH 7 phosphate buffer is added to the filtrate. The solution is evaporated under reduced pressure to 2 ml. and the mixture is taken up in 5 ml. of water and 5 ml. of ethylacetate and centrifuged. The ethylacetate layer is removed and the aqueous layer is extracted again with ethylacetate and with ether and then filtered through Celite. The aqueous solution is applied to a column (20 ml) of XAD-2 resin. The column is first eluted with water and then with 10% tetrahydrofuran. The tetrahydrofuran eluated is concentrated and lyophilized giving substantially pure O-methyl-Thienamycin.

EXAMPLE 34

Preparation of Thienamycin O-Phosphate Ester ##STR53##

Step A

To a solution of N-(p-nitrocarbobenzyloxy)-(Thienamycin-(p-nitrobenzyl)-Ester (50 mg) in 5 ml THF at 3.degree. C. is added 30 mg. of dibenzyl phosphorochloridate followed by 14 .mu.l of triethylamine. The mixture is stirred at 25.degree. C. for 2 hours. whereupon the THF is removed in vacuo. The residue is taken up in methylene chloride and washed with water. The methylene chloride solution is dried over magnesium sulfate and evaporated. The residue is chromatographed on silica gel yielding O-dibenzylphophoryl-N-benzyloxycarbonyl-Thienamycin benzyl ester.

Step B

A solution of the above product (20 mg.) in 10 ml. of 80% aqueous dioxane containing 8 mg. of NaHCO.sub.3 is hydrogenated in the presence of 20 mg. of 5% Pd on charcoal catalyst for 6 hours. The catalyst is removed by filtration and the filtrate evaporated to 2 ml. The solution is extracted twice with methylene chloride then concentrated and freeze dried, leaving the product Thienamycin O-phosphate disodium salt.

EXAMPLE 35

Preparation of O-(Methylcarbamoyl)-Thienamycin ##STR54##

A solution of N-(p-nitrobenzyloxycarbonyl)-Thienamycin-(p-nitrobenzyl) ester (20 mg.) and methylisocyanate (20 mg.) in methylene chloride (5 ml.) is stirred at 23.degree. C. for 18 hours. The solvent is evaporated and the residue extracted with hexane. The hexane insoluble residue is chromatographed on silica gel giving substantially pure O-(Methylcarbamoyl)-N-p-nitrobenzyloxycarbonyl-Theinamycin-p-nitrobenzyl ester. Following the hydrogenation procedure of Example 34, Step B, the product is obtained when the indicated substitution is made.

EXAMPLE 36

Preparation of O-(Methoxymethyl)-Thienamycin ##STR55##

Step A

A solution of 58 mg. of p-nitrobenzyloxycarbonyl-Thienamycin-p-nitrobenzyl ester in 5 ml of 1.0 ml of THF and NKPA is cooled to -78.degree. C. To this solution is added with stirring a 2 N solution of phenyllithium (0.1 ml.) immediately followed by the addition of 0.2 ml. of methylchloromethyl ether. The mixture is allowed to warm to 25.degree. C. during a period of one hour. Methylene chloride (25 ml.) is added and the solution is extracted with 0.1 N, pH 7, phosphate buffer (25 ml.), water 4.times.25 ml. The methylenechloride solution is evaporated and the residue is triturated with hexane. The hexane insoluble residue is chromatographed on silica gel yielding O-methoxymethyl-N-(p-nitrobenzyloxycarbonyl-Thienamycin-(p-nitrobenzyl)ester.

Step B

Following the hydrogenation procedure of Example 34, Step B, the title compound is obtained when the indicated substitution is made.

EXAMPLE 37

Following the procedures set out in the foregoing Examples and text, the following compounds of the present invention are obtained:

__________________________________________________________________________ ##STR56##Compound X R R.sup.1 R.sup.2__________________________________________________________________________(1.) O ##STR57## H ##STR58## (2.) O CH.sub.2 CH.sub.2 N(CH.sub.3).sub.2 H ##STR59## (3.) O ##STR60## H ##STR61## (4.) O ##STR62## H ##STR63## (5.) O ##STR64## H ##STR65##(6.) O ##STR66## H ##STR67## (7.) O ##STR68## H ##STR69## (8.) O CH.sub.2CH.sub.2CHCH.sub.2 H ##STR70## (9.) O CH.sub.2 CH.sub.2 N(C.sub.2 H.sub.5) H ##STR71## (10.) S CH.sub.2 CH.sub.3 H ##STR72## (11.) O CH.sub.2OCH.sub.3 H ##STR73## (12.) O ##STR74## H ##STR75## (13.) O SiC(CH.sub.3).sub. 3 H ##STR76## (14.) O CH.sub.2CHCH.sub.2 H SCF.sub.3 (15.) O CH.sub.2 CH.sub.2SCH.sub.3 H ##STR77## (16.) O ##STR78## H ##STR79## (17.) O ##STR80## H ##STR81## (18.) O ##STR82## H ##STR83## (19.) O ##STR84## H ##STR85## (20.) O ##STR86## H ##STR87## (21.) O ##STR88## H ##STR89## (22.) -- ##STR90## ##STR91##__________________________________________________________________________

EXAMPLE 38

Preparation of Pharmaceutical Compositions

One such unit dosage form consists in mixing 120 mg. of N-glycyl Thienamycin pivaloyloxymethyl ester with 20 mg. of lactose and 5 mg. of magnesium stearate and placing the 145 mg. mixture into a No. 3 gelatin capsule. Similarly, by employing more of the active ingredient and less lactose, other dosage forms can be put up in No. 3 gelatin capsules and should it be necessary to mix more than 145 mg. of ingredients together, larger capsules such as compressed tablets and pills can also be prepared. The following examples are illustrative of the preparation of pharmaceutical formulations:

______________________________________TABLET PER TABLET______________________________________N--glycyl Thienamycin pivaloyloxymethyl ester 125 mg.Cornstarch, U.S.P. 6 mg.Dicalcium Phosphate 192 mg.Lactose, U.S.P. 190 mg.______________________________________

The active ingredient is blended with the dicalcium phosphate, lactose and about half of the cornstarch. The mixture is then granulated with a 15% cornstarch paste (6 mg.) and rough-screened. It is dried at 45.degree. C. and screened again through No. 16 screens. The balance of the cornstarch and the magnesium stearate is added and the mixture is compressed into tablets, approximately 0.5 inch in diameter each weighing 800 mg.

PARENTERAL SOLUTION

Ampoule

N-glycyl Thienamycin pivaloyloxymethyl ester: 500 mg.

Diluent

Sterile Water for Injection: 2 cc.

OPTHALMIC SOLUTION

N-glycyl Thienamycin pivaloyloxymethyl ester: 100 mg. Hydroxypropylmethyl Cellulose: 5 mg. Sterile Water: to 1 ml.

OTIC SOLUTION

N-glycyl Thienamycin pivaloyloxymethyl ester: 100 mg. Benzalkonium Chloride: 0.1 mg. Sterile Water: to 1 ml.

TOPICAL OINTMENT

N-glycyl Thienamycin pivaloyloxymethyl ester: 100 mg. Polyethylene Glycol 4000 U.S.P.: 400 mg. Polyethylene Glycol 400 U.S.P.: 1.0 gram

The active ingredient in the above formulations may be administered alone or in combination with other biologically active ingredients as, for example, with other antibacterial agents such as lincomycin, a penicillin, streptomycin, novobiocin, gentamicin, neomycin, colistin and kanamycin, or with other therapeutic agents such as probenecid.

Claims

1. A compound having the structural formula: ##STR92## wherein R is a member selected from the group consisting of loweralkyl which may be substituted with loweralkoxy, mono- and diloweralkylamino, loweralkanoyloxy, loweralkanoylthio, loweralkanoylamino, loweralkylthio and pivaloxy; loweralkenyl; benzyl which may be substituted with loweralkyl, loweralkoxy, phenoxy, halo, nitro and pivaloxy; benzhydryl, phenyl, nitrophenyl, phenacyl and bromophenacyl; and R' is a member selected from the group consisting of: ##STR93## wherein R" is a member selected from the group consisting of hydrogen; loweralkyl (R" is not methyl) which may be substituted by hydroxy, loweralkoxy, phenyl, phenoxy, halo, amino, diloweralkylamino and guanidino; loweralkenyl, thionitrophenyl, benzyloxy, nitrobenzyloxy, loweralkoxybenzyloxy, loweralkoxy, haloloweralkoxy, loweralkylamino, loweralkylthio and thiophenyl.

2. The compound of claim 1 wherein R is pivaloyloxymethyl and R' is glycyl.

3. An antibiotic pharmaceutical composition consisting essentially of a therapeutically effective amount of a compound according to claim 1 and a pharmaceutical carrier therefor.

4. An antibiotic pharmaceutical composition consisting essentially of in unitary dosage form, a therapeutically effective amount of a compound according to claim 1 and a pharmaceutical carrier therefor.