IE46873B1 - Thienamycin derivatives - Google Patents

Abstract

Therapeutically active N-alkyl-N-iminomethyl derivatives of thienamycin of the formula I are made available. The meaning of R<1>, R<2>, R<3>, R<5>, R<6> and COXR<4> can be seen from Claim 1. These compounds are suitable as antibiotics. Processes for their preparation are also described.

Description

ted tc thienamycin, their preparation and pharmaceutical compositions containing them.

There is a continuing need for new antibiotics, for 5 there is unfortunately 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, known antibiotics suffer from the disadvantage of being effective only against certain types of microorganisms. Accordingly the search for new antibiotics continues.

The present invention provides novel antibiotics generically represented b y the following structural formula (I): 15 R θ I ® 1 2 SCH«CHON - C=NR R 22 Ifi C-X R which is one canonical form of a single resonant structure: C-X a k6 Formula I may more conveniently be represented by the symbol: Th ίί5 where Th symbolizes the bicyclic nucleus of thienamycin and the OH, amino, and carboxyl groups of thienamycin are illustrated. 2 In the foregoing formulae each of R and R , indep endently of the other, is a radical of the type defined for R, a hydrogen atom, or a nitro, hydroxyl, Cj__6 alkoxyl amino, 0^_6 alkylamino,di(C1_g alkyl)amino or tri(C1_g alkylamino) radical, an extra anion being present in the 2 latter case; or R and R are joined together to form, together with the nitrogen atom to which they are attached, a substituted or unsubstituted monocyclic or bicyclic heteroaryl or heterocyclyl residue containing 4-10 ring atoms, one or more of which may be an additional hetero atom selected from oxygen, sulphur and nitrogen? R is a cyano group or a substituted or unsubstituted carbamoyl, carboxyl, (cj_q0 alkoxy) carbonyl, alkyl, C2-lo alkenyl, c2_io alk7n71' C3-10 « C4_i2 cl'cloal-J<Y~ lalkyl, c5_12 cycloalkylalkenyl, c3_10 cycloalkenyl, C5_^2 cycloalkenylalkenyl, C4_12 cycloalkenylalkyl, Cg_1Q aryl, C7-16 a*311'!'1» Cg-16 a^alkenyl, Cg_j_g aralkynyl or monocyclic or bicylic heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalkyl comprising 4 to 10 ring atoms one or more of which is a hetero atom selected from oxygen, sulphur and nitrogen and in which the alkyl residue of the heteroaralkyl or heterocyclylalkyl radical contains from C to 6 carbon atoms; the substituent or substituents on R, io 12 ra, R or on the ring formed by joining R and R are chlorine; bromine; iodine; fluorine; azido; C^_4 alkyl; mercapto; sulpho; phosphono; cyanothio (-SCN); nitro;cyano; amino; hydrazino; amino or hydrazine having up to three C^_galkyl substituents; hydroxy; C^_g alkoxy; G^_g alkylthio; carboxyl; oxo; (C^g alkoxy)carbonyl; C2_1Q acyloxy? carbamoyl; (C^ alkyl)carbamoyl or di(C1_4 alkyl)carbao moyl; R is a hydrogen atom, an acyl radical or a radical d 4 of the type defined for R ; R is alkyl; substituted methyl carbonyl. i(C1_galkoxy)-(c1_6 . , \ alkyl);(C3_g cycloalkoxy) -(C^g alkyl);C2_12 alkanoyloxyalkyl;partially or completely halogenated C^_g alkyl in which the halogen(s) is/are chlorine, bromine or fluorine; C2-lo ai^enyi'· C3-i4 alkoxycarbonylalkyl; Cg_2^ dialkylaminoaoetoxyalkyl; ^2-13 alkaMYlffitti-noaIkyl; ar-(C^_3 alkyl) in which the aryl residue contains from 6 to 10 carbon atoms; monocyclic or bicyclic heteroaralkyl or heterocyclylalkyl containing '4 to' 10 ring atoms and 1 to 6 carbon atoms in the 2U alkyl residue, the hetero atom or atoms being oxygen, sulphur and/or nitrogen; nuclear-substituted aralkyl or heteroaralkyl in which the substituent is chlorine, fluorine, bromine, iodine or C^_g alkyl? aryl or nuclear-substituted aryl containing 6 to 10 ring carbon atoms and in which any nuclear substituent is hydroxy, alkyl, chlorine, fluorine, bromine or C2^2 alkylthioalkyl? C4_^2 cycloalkyl thioalkyl or (c2_j_0 acylthio)-(C^_g alkyl); R5 is substituted or unsubstituted C1_1Q alkyl; C2_1Q alkenyl or alkynyl; ring substituted and unsubstituted cycloalkyl, cycloalkenyl cycloalkenylalkyl, and cycloalkylalkyl having 3-5 ring carbon atoms and up to 6 carbon atoms in any chain; Cg_^Q aryl; aralkyl having 5-10 ring carbon atoms and 1-6 carbon atoms in the alkyl chain; monocyclic or bicyclic heteroaryl or heteroaralkyl containing 4-10 ring atoms, one or more of which is oxygen, nitrogen or sulphur, and 1-6 carbon atoms in the alkyl chain; and the ring or chain substituent (s) is/ are chlorine, bromine, iodine, fluorine, azido, cyano, amino, C^_g alkylamino, di (Cj_6 alkyl) amino or tri(C1__6 alkylamino) radical, an extra anion being present in the latter case, hydroxy, C^_g alkoxy, C3_g alkylthioalkyl; carboxyl; oxo, (C^_g alkoxy)carbonyl; Cj-lO acyl°xY' carbamoyl; (C1_4 alkyl)carbamoyl; di(C1_4 alkyl)carbamoyl; £ cyanothio (-SCN) or nitro; R is hydrogen, hydroxy, merca12 12 pto, R,-OR, -SR or NR R , where R, R and R are as defined above; 4 4 X is hydroxy, mercapto, amino, -OR , -SR , -NHR , -N-R , -0M, -0Q or, when the compound is in zwitterionic & form, -0 , in which case A is absent; A, when the compound is not in zwitterionic form, is a counter ion; M is a pharmaceutically acceptable cation; and Q is a blocking group as hereinafter defined. g In one preferred group cf compounds, R is C^_g 1 2 alkyl, each of R and R , independently of the other, is hydrogen or substituted or unsubstituted Cj__g alkyl, Cj_g alkenyl, C3_g cycloaikyl, C4_7 cycloalkylalkyl, C4_g cycloalkenyl, C4_7 cycloalkenylalkyl, C7_1Q aralkyl, Cg_10 aralkenyl, or monocyclic heteroar(C^_3 alkyl) having 5-6 ring atoms, one or more of which is oxygen, sulphur, and/ or nitrogen; in which the ring and/or chain substituent or substituetnts is or are chlorine, fluorine, hydroxy, c1-3 alkoxy, di(C^_3 alkyl)amino, or C^_3 alkylthio; the values hydrogen, Cj._g alkyl, C3_g alkenyl and benzyl, and especially hydrogen, methyl, ethyl, propyl, isopropyl, butyl, tbutyl, allyl and benzyl, being particularly preferred; and R6 is hydrogen; 0χ_6 alkyl; C2_g alkenyl; C2_g alkoxyalkyl; C2-12 alkylaittinoalkyl; C3 _32 dialkylaminoalkyl; C^_g perfluoroalkyl; ^2-6 alkylttu-°alkyl; or a substituted or unsubstituted phenyl; benzyl; phenethyl; monocyclic heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalkyl radical in which the substituted or substituents is or are chlorir?.fluorine, hydroxy, C1_3 alkoxy, di(C^_3 alkyl) amino or C-j__3 alkylthio; the values hydrogen, methyl, ethyl, propyl, isopropyl, methoxymethyl, hydroxyethyl, hydroxymethyl, dimethylaminomethyl, methylaminomethyl, trifluoromethyl, methyl thiomethyl,' ethylthiomethyl, phenyl, benzyl, phenethyl, 2pyridyl, 3-pyridyl, 4-pyridyl, 2-thiazolyl or 4-thiazolyl being particularly preferred.

In another preferred group of compounds, R5 is C^_g alkyl, R® is NR^R^ and each of R^ and , independently of the others, is as defined in the preceding paragraph.

Among specific compounds of the present invention are those of formulae where A is a pharmaceutically acceptable anionic salt residue as well as those individually mentioned below. 3 When R is acyl, the group OR is classifiable as an ester; when R is a group such as alkyl, aryl or aral6 kyl, the group OR is classifiable as an ether. The term acyl includes the alkanoyls including derivatives and analogues thereof such as thio analogues in which the carbonyl oxygen is replaced by sulfur; as well as sulfur and phosphorus acyl analogues such as substituted sulfonyl-, sulfinyl-, and sulfenyl- radicals, and substituted P(III and V) radicals such as substituted phosphorous, phosphoric , phosphonous and phosphonic radicals.

The nature of the counter anion, A, is not critical. It is representatively selected to provide pharmaceutically acceptable salts such as halides (e.g. chlorides or bromides), sulfate, phosphate, citrate, acetate or benzoate. Compounds of the present invention in which X is hydroxy are best considered to be zwitterions of the following structural representation (I)s OR R,5 COO6 + Thienamycin, the parent compound which may be used as a starting material in the preparation of the compounds of the present invention, is disclosed and claimed in U.S. Patent No. 3,950,357. It has the following structural formula; OH COOH This compound (including all isomers and mixtures of isomers thereof) is also available by the total synthesis which is described and claimed in British Patent Specifi0 II r* cation No. 1589896. This application makes available all isomers, pure and as mixture, of XI which are suitable starting materials for the preparation of the compounds of the present invention. Other convenient starting mater5 ials for preparation of the compounds of the present invent ion are N-alkylated thiensmycins and their 0- and carboxyl derivatives (III): OR3 -SCH2CH2NHR III wherein R3, X and R3 are as defined above. The N-alkylated thienamycins (III) are disclosed and claimed in British Patent Specification No. 1570988.

Reference may also be made to Patent Specification No. 44433, which discloses and claims N-iminomethyl derivatives of thienamycin (IV): OR -SCHoCH^-N^-NR-'-R2 2 j • 6 IV 12 where R , X, R and R are as defined above and their preparation from thienamycin. It should be noted that the preparation of compound IV from compound II is exactly analogous to the preparation of the compounds of the present invention I from compound III.

Finally, reference is made to Patent Specification No. 46877 , which discloses N-alkyl-N-acyl derivatives of thienamycin: sch2ch2-nr5r γ where R , X and R are as defined above and R' is acyl. Compounds IVa are required in the preparation of compounds g I when R is OR or SR.

It has been unexpectedly found that compounds of the present invention, which are characterized as N-alkyl-Niminanethyl derivatives of the antibiotic thienamycin, are broad-spectrum antibiotics and are useful in animal and human therapy and in inanimate systems. They are active against a broad range of pathogens which representatively include both gram-positive bacteria such as S. aureus, Strep, pyogenes and B. subtllls and gram-negative bacteria such as E. coli, Proteus morganli, Serratia, Pseudomonas and Klebsiella.

The compounds most usually prepared are those in c which R is alkyl or C2_g alkenyl, such as methyl, ethyl, propyl or allyl; benzyl; nuclear-substituted benzyl such as p-t-butyl benzyl or a heteroaralkyl such as 4pyridylmethyl, 2-furylmethyl or 2-thienylmethyl, Especially preferred compounds are those in which, in addition, X is hydroxy or OM, and R3 is hydrogen. In the subsequent description, the balancing ion A is omitted.

The compounds of the present invention are most conveniently prepared by derivatizing a suitably substitu- In these formulae, is C^_g alkyl. The ester of formula: X and R are as defined above and R compound may be treated with an imide NR1 II „ R - C - X°R'' an imidohalide of formula RRN = C-X' I R X°R* or a compound of formula R’R' 2 where R and R are as defined above, each X is a halogen atom, X° is oxygen or sulfur and -X°R*1 forms a leaving group. Alternatively, a compound of formula: in which X' is an -OR or-SR leaving group, is treated with 1 2 an amine NHR R to produce a compound of formula (I) in 6 12 which R is Cj__g alkyl and R is -NR R . For compounds of the present invention in which R® is OR or SR and R® is C^_6 alkyl, starting material IVa (above), in which the acyl radical R' is a carbamoyl or thiocarbamoyl radical of formula NR^R2-C -, where X° is oxygen or sulfur, is reacted X° with an alkylating agent or analogous R-containing agent calculated to provide a compound of formula: 6 8 73 As mentioned above, the reaction (III—>1 or IVa— —I) is perfectly analogous to the reaction (II—/IV) described in Patent Specification No. 44433 . except that starting material II (or its 0- and/or carboxyl derivatives) is replaced by an equivalent amount of starting material III (or IVa).

The starting material III and its preparation are fully disclosed and claimed in British Patent Specification No. 1,570988.

Such N-monoalkyl thienamycin derivatives are prepared by reacting thienamycin or a suitable derivative thereof or a suitably protected thienamycin compound with an N-alkylating agent. There is no undue criticality in the process and any of a variety of well-known N-alkylation procedures may be used. The identity of the N-alkylating agent is a matter of choice within the limits set by the definition of lA. The N-alkylation may be conducted in any of a variety of solvent systems that are inert or substantially inert to the desired course of reaction. Suitable solvents include polar solvents such as water, C, , alkanols such as JL-0 ethanol, dioxane, tetrahydrofuran (THF), acetonitrile, hexamethylphosphoramide (HMPA), dimethylformamide (DMF) and mixtures (particularly aqueous mixtures) of the above; and non-polar solvents such as benzene and halohydrocarbons such as methylene chloride and chloroform. Typically the reaction is conducted at a temperature of from -40°C to 50°C for from a few minutes to 5 hours. Usually, the react11 ν'-»’ ion is conducted in the presence of an acid acceptor such as propylene oxide, magnesium oxide or potassium carbonate. The preferred N-alkylating agents include active halides, sulfate esters, and Michael addition reagents. The following reagents are representative of such alkylating agents: methyl iodide, allyl bromide, bromoaeetone, phenacyl bromide, benzyl bromide, ethyl chloroacetate, propargyl bromide 2-bromoethyl ethyl ether, dimethyl sulfate, methylfluorosulphonate, chloromethylthiocyanate, chloroethylmethylsulfide, brcraanethylcyclcnrapar^, 2,4-dinitrofluorobenzene, 2chloromethylpyridine, acrylonitrile, methyl methacrylate and nitroethylene.

The starting material III, may be prepared in any of a variety of ways. One convenient starting material is tris-trimethylsilyl thienamycin /Th(TMS)37 (see below). 4 3 4 When it is desired for R , R or R and R to be other than hydrogen, suitably derivatized starting materials such as Ia, lb and Ic (below), may be used.

OR-* SCH2CH2NH2 -cox IC where R^ and X are as defined above and further exemplified below. Starting material Ia, which is also useful as an antibiotic, is claimed in British Patent Specification Nos. 1569234 and 1570987. These applications describe useful starting materials for preparing III, and, additionally, analogous processes for converting the N-alkylated N-iminomethyl thienamycins of the present invention to carboxyl-, 0-; and carboxyl- and O-derivatized forms that are also embraced by the present invention, which are also useful as antibiotics.

As stated above, the N-alkylation reaction is carried out in any of the above-named solvents in the presence ς of the N-alkylating agent of choice, such as R X', where X' is a leaving group such as halogen atom or a sulphate ester. When using, for example, the above-mentioned Th( TMS)^ then the desired product is obtained by aqueous hydrolysis following the N-alkylation step. The following reaction diagram summarizes the process: Th.

-OTMS -NHTMS COOTMS i—OTMS R5X' Th-NR5TMS COOTMS hydrolysis Th I-OH NHR5 I—COOH where TMS is trimethylsilyl, and R5 and X' are as defined above.

A second scheme for the preparation of mono-alkyl thienamycins (III) involves the N-alkylation of an N-substltuted thienamycin where the substituent is an easily removable bulky group (R°) such as aralkyl group, for example substituted and unsubstituted benzyl, benzhydryl (-CH(CgH5)2) and trityl (-C(CgHg)3) where the ring substituent on the aralkyl may be halo, nitro, alkyl or C1_g alkoxy. The following reaction diagram summarizes this scheme: -OR3r0R3 Th- —NHR° R5X' Th· -NR°R5 H2/cat. Th- - -COX -COX 1 2 3 r—OR -COX Th- 3 -OR -NR°R° -OR3 R5X'^ Th·—NR°R°R5 H,/cat Th--> Ί Lcox L-COX la where all symbols are as defined.

Xn words relative to the above diagram, starting material 1, prepared for example from the reaction of thienamycin or derivatives thereof with an aralkyl halide, is reacted with the N-alkylating agent of choice R3X', as above-described, to provide the Ν,Ν-dialkyl intermediate 2. The aralkyl N-substituent R° is readily removable to provide 3 by hydrogenolysis. Suitable conditions for this final cleavage step involve hydrogenating 2 in a solvent such as ethanol under hydrogen (1 to 4 atmospheres) in the presence of a catalyst such as platinum, palladium, or oxides thereof. The ultimate product of this reaction is primarily 3, the N-monoalkyl compound. However, there is some copresence of Ν,Ν-dialkyl thienamycin. Such contaminating by-products may be separated by chromatographic methods and the magnitude of contamination may be minimized by using ς one equivalent or less of the alkylating agent R X'.

A third method for the preparation of N-monoalkyl compound, III, particularly N-(C^_^Q alkyl) compounds, is similar to the above described procedure except that the starting material la is an Ν,Ν-diaralkyl-thienamycin. The preparation of such starting materials is described below. The following reaction diagram summarizes this process: i—OR3 NR5 I—COX III where all symbols are as described above. It is to be noted that this scheme for the preparation of N-(C^_3o alkyl) thienamycins is not complicated by the co-preparation of N,N-di(C1_10 alkyl) thienamycins.

A fourth method which is particularly suitable for the preparation of Ν-(0^_3θ alkyl) thienamycins species (III) involves the N-alkylation of a Schiff's Base of thienamycin. The following diagram illustrates and summarizes the reaction.

—OR·5 f~ORJ ΦΗ hydrolysis or - N=C-0 RX'^ Th—| - N=C-0 hydrogenation Th· ! H Is - Θ 1—cox —COX X' —OR COX 6 or III where all symbols are as previously defined and in addit4 3 ion φ is phenyl- or nuclear substituted phenyl, R and R may be the trlmethylsilyi radical (TMS) and X may be oxygen. The preferred Schiff's base is that obtained by reacting thienamycin with benzaldehyde or nuclear-substituted benzaldehyde. The process for preparing such Schiff's bases is not critical. Their preparation is disclosed in British Patent Specification No. 1570987 This copending application describes the preparation of starting material 4. The reaction of 4 with the c alkylating reagent R X' provides intermediate compound 5 which upon aqueous hydrolysis or catalytic hydrogenolysis provides the desired N-(C1_1Q alkyl)-thienamycin 6.

A fifth method for preparing N-(Cj_^0 alkyl) thienamycins (III) involves the desulfurization of an N-thioacyl thienamycin in the presence of a hydrogenation catalyst such as Raney Nickel: r—OR r-OR ThNHCR I! S -COX Raney Nickel Thnhch2r8 •COX o where X is hydroxy and R is as previously defined but are o preferably hydrogen, and R is hydrogen, aryl or alkyl.

The N-thioacyl thienamycin starting materials are fully disclosed in British Patent Specification No. 1 570 986.

The above desulfurization is typically conducted in polar protic solvents such as water, C^_g alkanols such as ethanol, and aqueous mixtures thereof, at a temperature of from 0-50°C for from 2 minutes to 5 hours.

Identification of the Radical -COX In the generic representation Of the compounds of the present invention (I, above), the radical represented by -COX, is, inter alia, -COOH (X is hydroxy) and all radicals known to be effective as pharmaceutically accept4 4 able salt, ester, anhydride {X includes R and R is acyl) and amide radicals in the bicyclic 0-lactam antibiotic art, such as the cephalosporins and penicillins and the nuclear analogues thereof.

Suitable radicals (R or Q) include conventional protecting or carboxyl blocking groups. The term blocking group as herein defined, is used in the same manner and in accordance with the teaching of O.S. Patent 3,697,515. 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 oossible ester groups, wherein X=OQ and Q is given: (i) Q= CRaRbRc where at least one of Ra, Rb and Rc Is an electron-donor, e.g., p-methoxyphenyl, 2,4,6-tri16 methylphenyl, 9-anthryl, methoxy, CHjSCH-j, tetrahydrofur2-yl, tetrahydropyran-2-yl or fur-2-yl. The remaining Ra, R and R groups may be hydrogen or organic substituting groups. Suitable ester groups of this type include p-methoxybenzyloxycarbonyl and 2,4,6-trimethylbenzyloxycarbonyl. (ii) Q= CRaRbRc where at least one of Ra, Rb and Rc is an electron-attracting group, e.g., benzoyl, p-nitrophenyl, 4-pyridyl, trichloromethyl, tribromomethyl, iodomethyl, cyanomethyl, ethoxycarbonylmethyl, arylsulphonylmethyl, 2-dimethylsulphoniummethyl, o-nitrophenyl or cyano. Suitable esters of this type include benzoyimethoxvcarbonyl, p-nitrobenzyloxycarbonyl, 4-pyridylmethoxycarbonyl, 2,2,2-trichloroethoxycarbonyl and 2,2,2-tribromoethoxycarbonyl . (iii) Q=CRaRbRc where at least two of Ra, Rb and Rc are. hydrocarbon such as alkyl, e.g., methyl or ethyl, or aryl, e.g, phenyl and the remaining Ra, Rb and Rc group, if there is one, is hydrogen. Suitable esters of this type include t-butyloxycarbonyl, t-amyloxycarbonyl, diphenylmethoxycarbonyl and triphenylmethoxycarbonyl. (iv) Q=R^, wherein R^ is adamantyl, 2-benzyloxyphenyl, 4-methylthiophenyl or tetrahydropyran-2-yl.

Silyl esters, under this category or blocking groups, may conveniently be prepared from a halosilane or a silazane of the formula: 41 41 41 41 4« R* 3SiX',· R 2SiX'2,· R 3Si.NR*2? R jSi.NH.COR* j R4'3Si.NH.CO.NH.SiR4'3; R4'nh.CO.NH .SiR4'3; or R4'c(OSiR43); 41 HN(SiR 3)2 wherein X* is a halogen such as chloro or bromo 41 and each R , which can be the same or different from the others represents hydrogen, alkyl, e.g., methyl, ethyl, n-propyl, iso-propyl; aryl, e.g., phenyl; or aralkyl, e.g., benzyl.

More geiierally stated, pharmaceutically acceptable carboxyl derivatives of the present invention are those derived by reacting thienamycin or an N-protected thienamycin such as III or an N-acylated thienamycin or a compound of the present invention I with such compounds as alcohols, phenols, mercaptans, thiophenols and acylating reagents which then may be derivatized to establish the R group of the compounds of the present invention (I, above). For example, esters and amides of interest are the compounds of formula I (above) having the following group at the 2-position: ★ -COXR4 * 4 where X is oxygen, sulfur, or NR’ (R' is H or R ), and R is alkyl having 1-10 carbon atoms, straight or branched, such as methyl, ethyl, t-butyl, pentyl or decyl;substituted carbonyImethyl, including phenacyl, p-bromophenacyl, £-t-butylphenacyl, acetoxyacetyImethyl, pivaloxyacetylmethyl, carboxymethyl, and its alkyl and aryl esters, and d-carboxy-a-isopropyl; aminoalkyl including 2-methylaminoethyl, 2-diethylaminoethyl, 2-acetamidoethyl, phthalimidomethyl and succinimidomethyl; alkoxyalkyl wherein the alkoxy portion has 1-6 carbons atoms; but can be branched straight or cyclic, and the alkyl portion has 1-6 carbon atoms, such as methoxymethyl, ethoxymethyl, isopropoxymethyl, ethoxypropyl, or cyclohexyloxymethyljalkanoyloxyalkyl where the alkanoyloxy portion is straight or branched and has 1-6 carbon atoms and the alkyl portion has 1-6 carbon atoms, such as acetoxymethyl, pivaloyloxymetbyl, acetoxyethyl, propionyloxyethyl or acetoxypropyl, halogenated alkyl where 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, trifluoro18 6 8 7 3 ethyl, 2-bromopropyl, diiodomethyl, 2-chloroethyl or 2bromoethyl; alkenyl having 2-10 carbon atoms, either straight or branched, e.g., allyl, 2-propenyl, 3-butenyl, 4- butenyl, 4-pentenyl, 2-butenyl, 3-pentenyl, 3-methyl-2butenyl, methallyl or 1,4-cyclohexadien-l-yl-methyl; alkynyl having 2-10 carbon atoms, either straight or branched, e.g., propargyl, ethynyl or 3-butyn-l-yl; alkanoyl, either straight or branched, having 1-10 carbon atoms, such as pivaloyl, acetyl or propionyl; substituted or unsubstituted aralkyl or heteroaralkyl having 4 to 10 ring atoms where alkyl has 1-3 carbon atoms, and hetero means 1-4 0,S or N hetero atoms, such as benzyl, benzhydryl, and substituted benzyl, benzhydryl, e.g. benzyl or benzhydryl substituted with 1-3 substituents such as halo, alkyl, or combinations thereof, e.g., p-chlorobenzyl, p-t-butylbenzyl, 2,4,6-trimethylbenzyl, or p-methylbenzyl, 2,2-dimethyl-5coumaranmethyl, 5-indanyImethyl, 2-thienyImethyl, 2-furylmethy.1, 3-t-butyl-5-isothiazolemethyl, phthalidyl; phenylethyl or 2-(p-methylphenyl)ethyl; aryl where aryl is phenyl, - indanyl, or substituted phenyl having up to 3 substituents, and is particularly unsubstituted phenyl or ortho or para monosubstituted phenyl, e.g., (4-methyl)phenyl, (4-hydroxy) phenyl or (4-t-butyl)-phenyl, phenyl alkenyl in which alkenyl has 2-6 carbon atoms, such as 3-phenyl-2-propenyl; aralkoxyalkyl where aralkoxy is benzyloxy and alkyl has 1-3 carbon atoms, such as benzyloxymethyl, 4-nitrobenzyloxymethyl, 4-chlorobenzyloxymethyl; alkylthioalkyl where the alkylthio portion has 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, methylthiopropyl, isopropylthioethyl or methyIthiobutyl.

In addition to the esters and thio esters listed above, amides are also embraced by the present invention, i.e., where X* is the -I- group.

Representative of such amides are those where R' is hydrogen, methyl, ethyl, phenyl, benzyl, or methylthioethyl, also embraced by -COX are anhydrides where X is benzyloxycarbonyloxy, ethoxycarbonyloxy, benzoyloxy or pivaloyloxy.

Especially preferred -COX radicals of the present invention are those in which (relative to Formula I above) * 4 , -ie X is X R where X is oxygen, sulphur or NR' (R' is hydrogen or alkyl); and R^ is C^g alkyl, C2_g alkenyl, such as methallyl, 3-methylbutenyl or 3-butenyl; methyl15 thioethyl; benzyl and substituted benzyl such as £-tbutylbenzyl; pivaloyloxymethyl, 3-phthalidyl and acetoxymethyl, propionyloxymethyl, acetylthiomethy1, pivaloylthiomethyl, allyl, 4-butenyl, 2-butenyl, 3-methyl-2butenyl, phenacyl, acetoxyacetylmethyl, methoxymethyl, -indanylmethyl, 5-indanyl, benzyloxymethyl, ethylthioethyl, methylthiopropyl, dimethylaminoacetoxymethyl, or acetamidomethyl. Among these, the particularly preferred compounds are those in which X is oxygen and R is hydrogen, methyl, t-butyl, phenacyl, £-bromophenacyl; pivaloyloxy25 methyl, 2,2,2-trichloroethyl, allyl, 3-methyl-2-butenyl, 2-methyl-2-propenyl, benzyl, benzhydryl, j>-t-butylbenzyl, phthalidyl, phenyl, 5-indanyl, acetylthiomethyl, acetoxymethyl, propionyloxymethyl, methallyl, 3-butenyl, 4pentenyl, 2-butenyl, acetoxyacetylmethyl, pivaloylacetyl30 methyl, dimethylaminoethyl, methoxymethyl, 5-indanylmethyl, benzyloxymethyl, methylthioethyl, dimethylaminoacetoxymethyl, acetamidomethyl, acetylthioethyl, pivaloylthiomethyl or methylthiomethyl.

Identification of R3 In the generic representation of the present 3 invention, structure I (above), the radical R is, in addition to hydrogen, 1.) acyl (generically the group ·> 3 -OR Is classifiable as an ester); or 2.) R is, for exa3 inple, alkyl, aryl or aralkyl such that the group -OR is 3 classifiable as an ether. For the esters (1) R is selected from the following definition of acyl radicals (p=l).

In the so-called ethers (2.) of the present invention, R is selected from the same acyl radicals in which the carbonyl residue, II -C-, or mere generally is deleted (p=0): thus R3 is selected from the following radicals where all symbols are defined below. 2 Thus, relative to the definition of R and R , the acyl radical can, inter alia, be substituted or unsubstituted alipphatic, aromatic or heterocyclic, araliphatic or heterocyclylaliphatic carboxylic acid radical, a substituted or unsubstituted carbamoyl radical or a carbothioic acid radical. One group of acyl radicals can be represented by the general formula: -C-R'' where X is 0 or S and R'' represents hydrogen; amino; substituted amino such as C^g alkylamino and di (C^g alkyl)amino; substituted or vnsubstituted straight or branched5 chain C^g alkyl; meroapto; aryloxy, typically comprising 6 to 10 carbon atoms; alk ery1 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 (monocyclic or hicyclic) where the alkyl residue typically comprises from 1 to 3 carbon atoms and the heterocyclic ring comprises typically from 4 to 10 atoms and the hetero atom or atoms is/are 0, N and/or S; such above-listed groups can be unsubstituted or can be substituted by radi15 cals such as OH, SH, SR (R is alkyl or aryl such as phenyl), alkyl or alkoxy groups having 1 to 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) in which the alkyl group(s) comprise(s) from 1 to 6 carbon atoms, C^_g haloalkyl such as trifluoromethyl, carboxy(C^_g alkyl), carbamoyl(C^_g alkyl) and N-substituted carbamoyl(C^_g alkyl), amidino, guanidino, N-substituted guanidino or guanidino-(C^_g alkyl). Representative examples of such acyl groups that might be mentioned are those in which R'' is benzyl, £-hydroxybenzyl, 4-aroino-4-carboxybutyl, methyl cyanomethyl, 2-pentenyl, n-amyl, n-heptyl, ethyl 3- or 4nitrobenzyl, phenethyl, ¢,6-diphenylethyl, methyldiphenylmethyl, triphenylmethyl, 2-methoxyphenyl, 2,6-dimethoxy30 phenyl, 2,4,6-trimethoxyphenyl, 3,5-dimethyl-4-isoxazolyl, 3-butyl-5-methyl-4-isoxaz olyl, 5-methyl-3-phenyl-4-isoxazolyl, 3-(2-chlorophenyl)-5-methyl-4-isoxazolyl, 3-(2,622 6 8 7 3 dichlorophenyl)5-methyl-4-isoxazolyl, D-4-amino-4-carboxybutyl, D-4-N-benzoylamino-4-carboxv-n-butyl, p-aminobenzyl, o-aminobenzyl, m-aminobenzyl, £-dimethylaminobenzyl, (3pyridyl)methyl, 2-ethoxy-l-naphthyl, 3-carboxy-2~quinoxalinyl, 3-(2,6-dlchlorophenyl)-5-(2-furyl)-4-isoxazolyl, 3phenyl-4-isoxazolyl, 5-methyl-3- (4-guanidinophenyl)-4isoxazolyl, 4-guanidinomethylphenyl, 4-guanidinomethylbenzyl, 4-guanidinobenzyl, 4-guanidinophenyl, 2,6-dimethoxy4- guanidino, o-sulfobenzyl, p-carboxymethylbenzyl, p-carbamoylmethylbenzyl, m-fluorobenzyl, m-bromobenzyl, p-chloroben2yl, £-methoxybenzyl, 1-naphthylmetiiyl, 3-isothiazolvlmethyl, 4-isothiazolylmethyl, 5-isothiazolylmethyl, guanylthiomethyl, 4-pyridylmethyl, 5-isoxazolylmethyl, 4-methoxy5- isoxazolylmethyl, 4-methy1-5-isoxazolylmethyl, 1-imidazolylmethyl, 2-benzofuranylmethyl, 2-indolylmethyl, 2-phenylvinyl, 2-phenylethynyl, 1-amlnocyclohexyl, 2- and 3-thienylaminomethyl, 2-(5-nitrofuranyl)vinyl, phenyl, o-methoxyphenyl, o-chlorophenyl, o-phenylphenyl, £-aminomethylbenzyl, 1-(5-cyanotriazolyl)methyl, difluoromethyl, dichlcromethyl, dibrcmarethyl, 1-(3-methylimidazolyl)methyl, 2- or 3-(5-carboxymethylthienyl) methyl, 2- or 3-(4carbamoylthienyDmethyl, 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-(4methoxy-1,2,5-thiadiazolyl)methyl, 2-furylmethyl, 2-(5nitrofuryl)methyl, 3-furylmethyl, 2-thienylmethyl, 3thienylmethyl, tetrazolylmet’nyl, benzamidinomethyl and cyclohexylamid-inomethyl.

The acyl group can also be a radical of the formula: X ll -C(CH2)nZR'' 6 8 7 3 where X is 0 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 -(CH2)nZR that might be mentioned are allylthiomethyl, phenylthiomethyl, butylmercaptomethyl, α-chlorocrotylmercaptomethyl, phenoxymethyl, phenoxyethyl, phenoxybutyl, phenoxybenzyl, diphenoxymethyl, dimethylmethoxyethyl, dimethylbutoxymethyl, dimethylphenoxymethyl, 4-guanidinophenoxymethyl, 4-pyridyl10 thiomethyl, £-(carboxymethyl)phenoxymethyl, £-(carboxymethylphenylthiomethyl, 2-thiazolylthiomethyl, £-(sulfo) phenoxymethyl, £-(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, trifluoromethylthiornethyl, 4-pyridylethyl, 4-;pyridylpropyl, 4-pyridylbutyl, 3-imidazolylethyl, 3-imidazolylpropyl, 3-imidazolylbutyl, 1-pyrrolo20 ethyl, 1-pyrrolopropyl, and 1-pyrrolobutyl.

Alternatively, the acyl group can be a radical of the formula: X -C-CHR'' R ' where R is defined as above and R ’ is a radical such as amino, hydroxy, azido, carbamoyl, guanidino, amidino, acyloxy, halo, suoh as Cl, F, Br, I, sulfamino, tetrazolyl, sulfo, carboxy, carbalkoxy or phosphono. Representative members of the substituent that might be mentioned are α-aminobenzyl, a-amino-(2thienyl)methyl, a-(methylamino)benzyl, a-amino-methylmercaptopropyl, a-amino-3- or 4-ehlorobenzyl, ct.-amino-3- or 4-hydroxybenzyl, a-amino-2 ,4-dichlorobenzyl, a-amino-3,4dichlorobenzyl, D(-)-a-hydroxybenzyl, α-carboxybenzyl, aamino-(3-thienyl)methyl D(-)-a-amino-3-chloro-4-hydroxybenzyl, α-amino(cyclohexyl)methyl, a-(5-tetrazolyl)benzyl, 2- thienyl-carboxymethyl, 3-thienyl-carboxymethyl, 2-furylcarboxymethyl, 3-furyl-carboxymethyl, α-sulfaminobenzyl, 3- thienyl-suifaminomethyl, a-(N-methylsulfaminoJ-benzyl D(-) -2-thienyl-guanidinomethyl, D(-)-α-guanidinobenzyl aguanylureidobenzyl, α-hydroxybenzyl, a-azidobenzyl, afluorobenzyl, 4-(5-methoxy-l,3-oxadiazolyl)-aminarethyl, 4{5-methoxy-l,3-oxadiazolyl)-hydro3$methyl, 4- (5-methoxy-l,3sulfadiazolyl) -hydroxymethyl, 4-(5-chlorothienyl) -aminaiethyl, 2- (5-chlorothienyl) -hydroxymethyl, 2- (5-chlorothienyl)-carboxy-methyl, 3- (1,2-thiazolyl)-aminomethyl, 3-(1,2-thiazolyl)hydroxymethyl, 3- (l,2“thiazolyl)-cartooxymethyl, 2-(1,4thiazolyl)-amincmethyl, 2- (1,4-thiazolyl)-hydroxymethyl, 2(1,4-thiazolyl)carboxymethyl, 2-benzothienvlaminomethyl, 2-benzothienylhydroxymethyl, 2-benzothienylcarboxymethyl, α-sulfobenzyl, a-phosphonobenzyl, α-diethylphosphono, and amonoethylphosphono. Further acyl radicals of interest in this class when X = oxygen are: ii 3 ' 4 ' -CCHR R 31 41 31 where R and R are as defined below. R represents hydrogen, halo, such as chloro, fluoro, bromo or iodo, amino, guanidino, phosphono, hydroxy, tetrazolyl, carboxy, sulfo or sulfamino and R3 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 or thiadiazolyl; substituted heterocycles, phenyIthio, phenyloxy C1-g alkyl, (heterocyclic or substituted heterocyclic)-thio ‘ 31 41 groups; or cyano. The substituents on R and R , can be halo, carboxymethyl, guanidino, guanidinomethyl, carboxamidomethyl, aminomethyl, nitro, methoxy or methyl. When 3' . 4 · R is hydrogen., hydroxy, amino or carboxy and R is phenyl or a 57 or 6-membered heterocyclic ring having one or two sulfur, oxygen and/or nitrogen hetero atoms such as tetrazolyl, thienyl, furyl and phenyl, the following acyl radicals are representative: phenylacetyl 3-bromophenylacetyl, £-aminomethylphenylacetyl, 4-carboxymethylphenylacetyl, 4-carboxyamidomethylphenylacetyl, 2-furylacetyl, 515 nitro-2-furylacetyl, 3-furylacetyl, 2-thienylacetyl, 5chloro-2-thienylacetyl, 5-methoxy-2-thienylacetyl, aguanidino-2-thienylaeetyl, 3-thienylacetyl, 2-(4-methylthienyl)acetyl, 3-isothiazolylacetyl, 4-methoxy-3-isothiazolylacetyl, 4-isothiazolylacetyl, 3-methyl-4-isothiazolyl20 acetyl, 5-isothiazolylacetyl, 3-chloro-5-isothiazolylacetyl, 3-methy1-1,2,5-oxadiazolylacetyl, 1,2,5-thiadiazolyl4- acetyl, 3-methy1-1,2,5-thiadiazolylacetyl, 3-chloro-l,2, - thiadiazolylacetyl, 3-methoxy-l,2,5-thiadiazolylacetyl, phenylthioacetyl, 4-pyridylthioacetyl, cyanoacetyl, 1-tet25 razolylacetyl, α-fluorophenylacetyl, D-phenylglycyl, 4hydroxy-D-phenylglycyl, 2-thienylglycyl, 3-thienylglycyl, phenylmalonyl, 3-chlorophenylmalonyl, 2-thienylmalonyl, 3thienylmalonyl, a-phosphonophenylacetyl, α-amino cyclohexadienylacetyl, α-sulfaminophenylacetyl, a-hydroxyphenyl30 acetyl, α-tetrazolylphenylacetyl and α-sulfophenylacetyl.

The acyl radical may also be selected from sulphur (1) and phosphorus (2) radicals; (0)m II (X)n | II -S-Y II 1 -P-Y I II (0)n 1 Y' ' where with respect to 1, each of m and n is 0 or 1 and Y is Οθ M®, -N(R)2, or R' '; where M® is hydrogen, an alkali metal cation or an organic base; and R'' is as defined above, e.g., alkyl, alkenyl, aryl or heteroaryl. With respect to 2 X = 0 or S; n=0 or 1; and each of Y’ and Y is 0θ M®, -N(R'')2, R'' or ZR'’ where all symbols are as defined above, e.g., R'' and ZR'' are representatively alkyl, alkenyl, aryl or heteroaryloxy, Y' and Υ'1, including R'1 residues, can be joined together to form cyclic ester, esteramide, and amide functions. Illustrative examples of the radicals 1 are methylsulphonyl, pnitrophenylsulphonyl, p-chlorophenylsulphinyl, o-nitrophenylsulphenyl, sulfamoyl, dimethylsulphamoyl and sulpho. Illustrative examples of the radicals £ are: dimethoxyphosphino, dibenzyloxyphosphino, dihydroxyphosphino, dimethoxyphosphinyl, dimethoxyphosphinothioyl, dibenzyloxyphosphinyl, dihydroxyphosphinyl.

An acyl class of particular interest comprises 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-butoxyoarbonyl and phenoxyacetyl; non-acyl protective groups such as tri(C1_6 alkyl) silyl, for example, trimethylsilyl and jt-butyldimethylsilyl are also of interest. 6 8 7 3 ! ι ! ι The following radicals, according to the foregoing definition of acyl, are preferred; formyl, acetyl, propionyl, butyryl, chloroacetyl, methoxyacetyl, aminoacetyl, methoxycarbonyl, ethoxycarbonyl, methylcarbamoyl, ethylcarbamoyl, phenylthiocarbonyl, 3-aminopropionyl, 4aminobutyryl, N-methylaminoacetyl, N,N-dimethylaminoacetyl, an Ν,Ν,Ν- trimethylaminoacetyl salt, 3-(Ν,Ν-dimethyl)aminopropionyl, a 3-(Ν,Ν,Ν-trimethyl)aminopropionyl salt, an Ν,Ν,Ν-triethylaminoace tyl salt, a pyridiniumacetyl salt, guanyIthioacetyl, guanidinoacetyl, 3-guanidinopropionyl, N -methylguanidinopropionyl, hydroxyacetyl, 3-hydroxypropionyl, acryloyl, propynoyl, malonyl, phenoxycarbonyl, amidinoacetyl, acetamidinoacetyl, amidinopropionyl, acetamidinopropionyl, guanylureidoacetyl, guany lcarbamoy 1, carhoxymethylaminoacetyl, sulfoacetylaminoacetyl, phosph3 onoacetylaminoacetyl, N -dimethylaminoacetamidinopropionyl, ureidocarbonyl, dimethylaminoguanylthioacetyl, 3-(l-methyl4-pyridinium)propionyl, 3-(5-aminoimidazol-l-yl)propionyl, a 3-methyl-l-imidazoliumacetyl salt, 3-sydnonylacetyl, o-sminarethylbenzcyl, o-aminobenzoyl, S s -p(och3)2, ICH.

-P.

OM, e.g S ONa -Ρ-Ν(ΟΗ,),, e.g. -P-N(CH,)-, \ 3“ \ OM ONa I -pJF(ch3 )f72, s s -P-N(CH,)9, e.g.-P-N(CH-),, \ \ OM ONa I -S-OM or II -p.

OM where M is hydrogen or an alkali metal or alkaline-earth metal cation.

An especially preferred class of acyl radicals are terminally substituted acyls in which the substituent is a basic group such as substituted or unsubstituted amino, amidino, guanidino, guanyl and nitrogen-containing mono10 cyclic and bicyclic heterocycles (aromatic and non-aromatic) where the hetero atom car atoms, in addition bo nitrogen, can be oxygen and/or sulfur. Such preferred substituted acyls may be represented by the following formula: -C(CH2)g-(A)p-(CH2)r-Y where each of 3 and r is 0, 1, 2, 3, 4 or 5; £ is 0 or 1; A is -0-, -NH-, N(C1_6 alkyl)- or -S- and Y is (1) substituted or unsubstituted amino or ammonium group of formula -N(R)2 or -N(R)3; an amidino or substituted amidino radical of formula -N=C-N(R)2 ; I R a guanidino or substituted guanidino radical of formula -NH-C-N(R)- ; Ii 2 NR or a guanyl and substituted guanyl radical of formula -C=NR NCR)2 ; where each R, independently of the others, is hydrogen; N(R')2 (R' is hydrogen or C^_g alkyl); C^_g alkyl, C^_g alkoxyl, (C^_g alkoxy)-(C2_6 alkyl), C3_g cycloalkyl, (C3_6 cycloalkyl)-(C1-3 alkyl), or (but only when R is attached to carbon), (C^_g alkoxy)methyl; or two R groups are joined to form, together with the atom(s) to which they are attached, a ring having 3-6 atoms: or (2) a nitrogen-containing monocyclic or bicyclic aromatic or non-aromatic heterocycle having 4 to 10 nuclear atoms in which the hetero atom(s), in addition to nitro10 gen, if any, is/are oxygen and/or sulfur. Such heterocycles are representatively illustrated by the following list of radicals, where R' is H or C^g alkyl having 1-6 carbon atoms: The following specific acyl radicals, in the formulae of which Q is an anion, which fall within this class are additionally representative and are preferred: NH H II -CCH2CH2NHC-CH3 NH -CCH2CH2NHC-H NH II I -CCH2CH2NHC-NH2 -CCH2CH2N(CH3)2 . ji -CCH2CH2N(CH3)3.q Q NH Ϊ H -CCH2CH2C-NH2 UH -cck2ch2ch2nhc-ch3 II © -CCH2CH2CH2N(CH3)3 .Q -CCH2CH2CH2N(CH3)2 A'H -CCH-S-C ΧΝΗ„ -CCH2S-CH2· II ^NH -CCH.-O-CH-C 2 2 Ah, 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. Preparation of Starting Materials Ia, lb, and Ic The above-described starting materials are conveniently prepared from an N-protected thienamycin species such as an N-acylated thienamycin (1) rOH Th- -NHR' 2' ‘-COOH fv wherein R* is an easily removable blocking group such as: carbobensyloxy, ring-substituted carbobenzyloxy such as o- and j3-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 ss triloweralkylsilyl, for example, trimethylsilyl, and t-butyldimethylsilyl are also of interest. The most preferred N-blocking groups are the substituted and unsubstituted carbobenzyloxy radical: where n is Ο, 1 or 2 and R’ is Cj__g alkoxy or nitro; and bromo-t-butoxycarbonyl. - 34 46873 The ultimate N-deblocking procedure for the preparation of la, lb or Ic is accomplished by any of a variety of well-known procedures which include hydrolysis or hydrogenation,- when hydrogenation is employed suitable conditions involve a solvent such as a loweralkanoyl in the presence of a hydrogenation catalyst such as palladium, platinum or oxides thereof.

The N-acylated intermediate (1, above) is prepared by treating thienamycin (II) 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 g-nitrophenyl ester. Such JIacylated thienamycin starting materials are fully described in above-cited, British Patent Specification No. 1570936.

The acylation reaction may be conducted at a temperature in the range of from about -20° to about 100°C., but is preferably conducted at a temperature in the range of from -5°C. to 25®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 phsophoramide (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 maxinum 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.

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 or acetonitrile in the presence of a suitable acceptor base such as NaHCO., MgO, NaOH or K2HPO4· In carrying out the reactions described herein, it is generally not necessary to protect the 2-oarboxy group or the l’-hydroxy group; however, in oases where the acylating reagent is exceedingly wafer-sensitive it is sometimes advantageous to perform the acylation in a non·' aqueous solvent system. Triorganosilyl (or tin) derivatives - 36 46873 of thienamycin proceed . rapidly to give the tris-triorganosilyl derivative, for example tris-trimethylsilyl thienamycin Th(TMS)3: r OTMS Th.

NHTMS <— COOTMS 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 25eC., with vigorous stirring under a N2 atmosphere. The resulting NH^Cl is removed by centrifugation and the solvent, is removed by evaporation to provide the desired silyl derivative.

The intermediate starting materials lb are prepared according to the following scheme; however, it should be noted that direct esterification, without protection of the amir.o group, is also possible. rOH rOH rOH Th--NHR2 -COOH Th· •nhr2 ' •cox lb deblock Th--NH2 -COX lb wherein all symbols are as previously defined.

In general, the transformation U.—>lb} is accomplished by conventional procedures known in the art. Such procedures include: 1. ) Reaction of 1 (or II,thienamycin) with a dia2oalkane such as diazomethane, phenyldiazomethane or diphenyldiazomethane in a solvent such as dioxane, ethylacetate or acetonitrile at a temperature of from 0*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, 68 73 or m-phenoxybenzyl bromide, p-t-butylbenzyl bromide, or pivaloyloxymethyl chloride. Suitable reaction conditions include solvents such as hexamethylphosphoramide at a temperature of from 0°C. to 60°C for from a few minutes to 4 hours. 3. ) Reaction of 1 with an alcohol such as methanol, ethanol or benzyl alcohol. This reaction may be conducted In the presence of a carbodiimide condensing agent such as dicyclohexylcarbodiimide. Suitable solvents,, at a temperature of from 0°C to reflux for from 15 minutes to IS hours, include CHC13, CH3G1 and CH2C12· 4. ) Reaction of an N-acvlated acid anhvdride of 1 * “ prepared by reacting the free acid £ with an acid chloride such as ethyl ehloroformate or benzyl chloroformate 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 ^1 and the acid chloride in a solvent such as tetrahydrofuran (THF) or CH2C12 at a temperature of frdm 25°C., to reflux· for from 15 minutes to 10 hours.

.) Reaction of labile esters of 1 such as the trimethylsilyl ester or dimethyl-t-butylsilyl ester v/ith R X' wherein X' is halogen such as bromo and chloro andR4 is as defined, in a solvent such as THF or CHjCl^ at a temperature of from O°C to reflux for from 15 minutes to 16 hours. For example according to the following schema: OTMS rOTMS “OTMS esterrThj-NHTMS H-agylation χ Th+NR2 TKS Th--NR2*TMS -COOR4 tCOOTMS OTOTMS -OH mild hvdrolvciκ.

Tl) - NHR2' -CGOR4 wherein TMS is triorganosilyl such as trimethylsilyl and all other symbols are as previously defined.

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

The above-recited schemes of esterification are 10 well known in the related bicyclic β-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 lb useful as starting materials in the practice of the present invention.

Starting materials la and Ic are conveniently prepared by any of a variety of well-known esterification or etherification reactions upon the secondary alcoholic group of lb Such procedures includes -OH Th--NHR2' COX 1.) For the preparation of ethers Of the present invention, the acid-catalysed reaction of lb with a diazoalkane such as diazomethane, phenyldiazomethane, or diphenyldiazomethane in an inert solvent such as dioxane, tetrahydrofuran (THF), halohydrocarbons such as CHjClj and ethyl acetate in the presence of a catalytic amount of a strong acid or Lewis acid such as f toluenesulfonic acid, trifluoroacetic acid, fluoroboric acid, or boron trifluoride at a temperature of from -78eC to 254C for from a few minutes to 2 hours. 2.) For the preparation of ether embodiments of the present invention, the·reaction of lb with an alkylating agent such as active halides, for example methyl iodide, benzyl bromide or m-phenoxybenzyl bromide; alkylsulphates such as dimethylsulphate and diethylsulphate or methylfluorosulphonate in the presence of a strong base capable of forming the alcoholate anion of lb. Suitable bases include alkali and alkaline-earth metal oxides I and hydrous oxides, alkali metal alkoxides such as potassium, tertiarybutoxide, 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) and dioxane at a temperature of from -78°C to 2S4C., for from a few minutes to 4 hours. 3.) por the preparation of ester embodiments, of the present invention, the reaction of lb with any of /V* the above-listed acyl radicals in their acid form. This reaction may be conducted in the presence of a carbodiimide condensing agent such as dicyclohexylcarbodiimide .

Suitable solvents include any inert solvent such as - 40 46873 CHClj, CH2Cl?, DMF, HMPA, acetone and dioxane at a temperature of from 0°C to 60“C. for from 15 minutes to 12 hours. 4. ) For the preparation of ester embodiments of the present invention, the reaction of lb 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 or DMF, in the presence of a suitable acceptor base such as NaHCOj, MgO, triethylene or pyridine, at a temperature of from C“C. to 40°C for from 1 to 4 hours.

Suitable acyl halides and anhydrides include: acetic anhydride, bromoacetic anhydride, propionic anhydride, benzcylchloride, phenylacetyl chloride,azidoacetyl chloride, 2-thienylacetyl chloride, 2-, 3- and 4nicotinyi chloride, p-nitrobenzoyl chloride, 2,6dimethoxvbenzoyl chloride, 4-guanidinophenylacetyl chloride hydrochloride, methanesulfonyl chloride, dibenzylphosphorochloridate, dimethylthiophosohorochloridate, 2-furoyl ethyl carbonic anhydride, methyl chloroformate and bis(p-nitrobensyl)phosFhorochloridate.

, ) For the preparation of ester embodiments of the present invention, the reaction of lb with a suitably substituted ketene or isocyanate such as ketene, dimethyl ketene, methyl isocyanate, methyl isothiocyanate, or chlorosulfonyl isocyanate.- Suitable solvents include dioxane, tetrahydrofuran and chloroform at a temperature of from -70’C. to 60®C. for from 15 minutes to 18 hours.

The intermediate compound / is then N-deblocked as described above to provide starting material Ic.

From Ic,. Ia is prepared by deblocking the carboxyl group: [“OR3 -OR3 -OR3 Th· - NHR2'— -> Th- -NH2 -> Th- -nh2 -COX -COX' .COOH 2 Ic la /V Starting material Ia is conveniently and preferably obtained when 4 4 X xs OR , where R is a removable carboxyl -protecting or blocking group (see above). Starting material Ia is prepared by deblocking according to any of a variety of well-known procedures which include hydrolysis and hydrogenation . When the preferred carboxyl-blocking groups are employed (below), the preferred deblocking procedure is hydrogenation, wherein the intermediate species (Ic or 2) in a solvent such as a C^_g alkanol, is hydrogenated in the presence of a hydrogenation catalyst such as palladium, platinum or oxides thereof.

In this connection, it is noted that suitable blocking groupsinclude 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 blocking groups R^ include benzyl, phenacyl, p-nitrobenzyl, methoxymethyl, trichloroethyl, trimethylsilyl, tributyltin, p-methoxybenzyl, benzhydryl. These blocking groups are preferred since they are generally recognized easily-removable blocking groups in cephalosporin and penicillin art.

The preferred carboxyl blocking groups are benzyl and substituted benzyl: wherien n is 0-2 (n«=0, R’«H) and R' is loweralkoxyl or nitro.

Preparation The preparation of the compounds of the present invention I is conveniently described according to the definition of R®. There are three cases: 1.) Amidines (R6»H, or R) ; 2.) Guanidines (R6=NRXR2) ; and 3.) Substituted Pseudoureas (R6»OR, or SR); wherein all symbols are as defined above. 1.} Amidines. In general the compounds of Class 1.) may conveniently be prepared by reacting the desired N-alkyl thienamycin (III) with an imido ester (a.) or a substituted imido halide (b.): a.) b.) 2 wherein R , R , and R are as defined above? X' is halo such as chloro; and -OR is a leaving group where R is C^_g alkyl such as methyl or ethyl.

Suitable solvents for the preparation of the compounds of Class 1 according to the above reaction schemes, depending upon the identity of the thienamycin substrate and reagent, include water, dioxane, tetrahydro 5 furan (THF), dimethylformamide (DMF), chloroform, acetone, acetonitrile or mixtures thereof. The reaction is conducted at a temperature of from 0’ to about 25°C for from 1 to about 6 hours. There is no criticality as to the precise identity of the reaction solvent nor the 10 variables of reaction within the limits described above, provided only that the reaction solvent is inert or substantially inert to the intended course of reaction. Suitable reagents representatively include: a.) Imido Esters: rV ©» R-C-X’R Rl X = 0 or S Methyl formimidate, ethyl formimidate, methyl acetimidate ethyl acetimidate, methyl benzimidate, ethyl 4-pyridyl carboximidate, methyl phenylacetimidate, methyl 3-thienyl· carboximidate, methyl azidoacetimidate, methyl chloroacetimidate, methyl cyclohexylcarboxiaidate, methyl 244 4 S 8 7 3 furylcarboximidate, methyl p-nitrobenzimidate, methyl 2,4dimethoxybenzimidate, ethyl N-methyl formimidate, methyl umethyl formimidate and methyl N-isopropyl formimidate.

Such imido ester reagents (a.) are conveniently prepared by any of a variety of known procedures, such as: 1. ) The reaction of a nitrile, RCN, with a g alkanol in the presence of HC1 according to the well-known Pinner synthesis. 2. ) The reaction of a nitrile, RCN, with a C. , alkanol in the? presence of a base. Typically, the 1—6 reaction is conducted at 0-40°C in the presence of an excess of the alcohol with a catalytic amount of an alkali metal alkoxide for from 15 minutes to 4 hours. n b 3. ) The reaction of an amide, RCNHRA, with an alkyl chloroformate, such as methyl chloroformate at 254C-45°C for 1-4 hours. 4. ) The reaction of an N-substituted amide, 0 Q ’12 RCNHR or RCN2 R , with an equivalent of an alkylating agent such as triethyloxonium fluoroborate in an inert solvent such as ether or chloroform at O-23°C for from 10 minutes to 2 hours.

) The conversion of a readily available imido ester, kCNR' (R1 mav be hvdrogen), to OR a desired imido ester, RCNR·*· , by reaction of the firstOR mentioned with an alkylaaine, R'NH2, in a mixture of water and ar. immiscible solvent such as ether or chloroform at 0-23C for from 5 minutes to 1 hour. b.) ‘Substituted Imido Halides: Chloropiperidino methylium chloride, chlorodimethylforminium chloride and chlorodiethyl forminium chloride.

Such imido halide reagents (b.) are conveniently prepared by any of a variety of known procedures, such as: 1.) The reaction of an Ν,Ν-disubstituted amide, R^NR^-R^ , with a halogenating agent such as thionyl chloride, 1q phosgene or phosphorus pentachloride in an inert solvent such as chloroform or methylene chloride at 0-40*C for from 1-5 hours.

The reaction may be representatively involving the reagents (a.), shown by the following diagram: where OR is the leaving group of 'the imido ester reagent and R, R1, R3 and X are as defined above.

This reaction is particularly suitable for embodiments wherein R3 and R4 are hydrogen and X is oxygen.

The reaction involving the reagents, (b.), may representatively be shown by the following diagram; ^R2N+»CX'_] X'^ III__> ? © 12 SCH2CH2N=C-NP.Air COXR . τ® mild hvdrolvsisx pH 3-6 ' * /V where all symbols are as previously defined. When product 2, is desired, suitable values for R3 and X are trimethylsilyl and trimethylsilyloxy, respectively. 1 2.) Guanidines: In general, the compounds of Class 2. may conveniently be prepared by reacting III with: (a.) an -OR (e.g., O-alkyl, O-aryl) pseudourea or an S5 alkyl or S-aryl pseudothiourea,· or (b.) by reacting a compound of Class 3. (above) with ammonia or an amino compound such as an alkyl, aralkyl or heteroaralkyl amine.

Suitable solvents for such reactions include water and buffered agueous polar organic solvent mixtures at pH 7-3 or anhydrous polar organic solvents such as dimethylformamide or hexamethylphospnoramide at a temperature of from 0°C. to 40'C. for from 1 to 24 hours. Suitable reagents, (a.) and (b.), include: (a.) -OR psnudoureas and -SR pseudothioureas: Ο-Methyl pseudourea, S-Methylpseudothioursa, S-methylpseudothionitrourea, 0-2,4-dichlorophenyl pseudourea, S-ja-nitrophenyl pseuuothiourea and o-N,N-trimethyl pseudourea, (b.) Amino reagents: 2q Methylamine, ethylamine, 2-arninopyrimidine, dimethylamine, methyl benzylamine, 3-aminomethyl pyridine, 2- aminomethyl thiophene, ethanolamine, dimethylaminoethylamine, N-2-(aminoethyl)pyrrolidine, eyclohexylamine, nheptylamine, isopropylamine, 2-rnethylallylamine, 3-phonyI25 1-propylamine, 2-amino-4-picoline, 2-amino-pyridine, 3- amino-4-carbethoxypyrazcie, 2-aminothiazole, S-amino-3methyl isothiazole, and 3-amino-l,2,4-thiazole.

The reaction involving the reagents, (a.) , 12 wherein R , X , R and R are as defined above; X is 0 or S and R” is as defined and preferably is lower alkyl or aryl.

The reaction involving the may representatively be shown by the reagents, (b.), following diagram; κ R- + SCH-CH-N^C-NpAr2 2 2 , - COX X’R OR3 Compound of Class 3. Λ . , —/>,—GCHnCH-lfe'C-NR^R2 HNRXR2 wherein X” is 0 of S and all other symbols are as previously defined. 3.) Suastitutea Pseudoureas; In geheral, the compounds of Class 3. may conveniently be prepared by reacting a carbanpyl or thiocarbanoyl N-substituted Thienamycin species IVa, (see above) for example; with an alkylating agent (b.) such as an active alkyl 10 or aralkyl halide or sulfate ester. - 50 468 73 Suitable media for the above reaction include C-p_g alkanols, dioxane and acetonitrile. The temperature may be from 20°C. to 60°C. and the reaction period from 1 to 4 hours.

Suitable reagents IVa for above reaction scheme I 1 ’ wherein RJ, R and X are as defined above and R is acyl as defined above and preferably is -?-NR^R2 or -CNR1R2 (R^ and R2 are as defined above), such as: carbamoyl, methylcarbamoyl, ethylcarbamoyl, phenylcarbamoyl, g-bromophenylcarbamoyl, phenylthiocarbamoyl, methylthiocarbamoyl and dimethylcarbamoyl.

Suitable reagents (b.), alkylating agents, include: methyl iodide, benzyl bromide, dimethylsulfate, diethylsulfate, allyl bromide, 2-thienyl bromide, methallyl bromide, g-nitrobenzyl bromide and methyl chloromethyl ether.

The reaction involving the above reagents IVa and (b.) may representatively be shown by the following diagram: wherein X is 0 or S; X is halogen such as bromo, iodo or alkyl sulphate; RX° is the alkylating agent; and R1, R$, 3 3' R , R , X, XJ, B and R are as previously defined.

Tne products of tnis invention (*ϊ) form a wide variety of pharmacologically acceptable salts such as acid addition salts, e.g., with hydrochloric, hydrobromic, .sulfuric, nitric , g-toluenesulfonic and methanesulfonip acids. The salts of the compounds of this invention are pharmacologically acceptable non-toxic derivatives which can be used as the active ingredient in suitable unitdosage pharmaceutical forms. Also, they may be combined witn other drugs to provide compositions having a broad , spectrum of activity.

The novel compounds are valuable antibiotics active against various gram-positive and gram-negative bacteria and, accordingly, find utility in human and veterinary medicine. 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 coll, Klebsiella pneumoniae, Serratia, Salmonella typhosa, Pseudomonas and 3acterium proteus. The antibacterials of the invention may further be utilized as additives to animal feeding stuffs, for preserving foodstuffs and as disinfectants. Por example, they may be included in aqueous compositions in concentrations ranging from 0.1 to 100 parts of antibiotic per million parts of solution in order to destroy ana inhibit the growth of harmful bacteria on medical and dental equipment- and as bactericides in industrial applications, for example, in water-based paints and in tne white water of paper mills to inhibit the growth of harmful bacteria.

X 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. They will normally be presented in a composition that also contains a suitable pharmaceutical carrier, preferably in unit dosage form containing a therapeutically effective amount of the antibiotic. Thus, the antibiotics and their corresponding salts may be presented as capsules or pills, tablets, in powder form, as liquid solutions or as suspensions or elixirs. They may be administered orally, intravenously or intramuscularly. The compositions may also contain an antibiotic and/or therapeutic agent that is not a compound of Formula I.

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, maize-starch, 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 preparations may be in the form of aqueous or oily suspensions, solutions, emulsions, syrups or elixirs, or may be presented as a dry powder 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, methyl cellulose, glucose/sugar syrup, gelatin, hydroxyethylcellulose, carboxymethyl cellulose, aluminium 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 £-hydroxybenzoates or sorbic acid. Suppositories will contain conventional suppository bases, e.g., cocoa butter or other glyceride.

Compositions for injections may be presented in unit dose form in ampoules, or in multidose containers with an added preservative. The compositions may take such forms as suspensions, solutions, or 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 dry powder form for reconstitution with a suitable liquid, e.g., sterile pyrogen-free water, before use.

The composition 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 and throat paints. For medication of the eyes or ears, the preparations may be presented as individual ophthalmic or otic capsules, in liquid or semi-solid form, or may be used as ophthalmic or otic liquids, e.g., in the form of drops. Topical applications may be formulated in hydrophobic or hydrophilic bases as ointments, creams, lotions, paints, powders, and semisolids.

Also, in addition to a carrier, the compositions may include other ingredients such as stabilizers, binders, antioxidants, preservatives, lubricators, suspending agents, viscosity agents or flavoring agents. In addition, there may also be include d 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 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 2 to 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 15 to 150 mg, of active ingredient per kg. of body weight.

The 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 93% of active material, the preferred range being from about 10-60%. The composition will generally contain from 15 mg. to 1500 mg. of fchfi active Ingredient; however, in general, it is preferable that the dosage be in the range of from 100 mg. to 1000 mg. IW parenteral administration the unit dosage is usually the pure compound in a slightly acidified sterile water solution or in the form of a soluble powder intended for solution.

In the following examples which illustrate but do not limit the present invention, the words Celite, Dowex”, Nujol and Porasil are Trade Marks and temperatures are in °C.

EXAMPLE 1 Preparation of N-Methyl-N-Formimidoyl thienamycin NCE CH.

NH Π N-raethyl thienamycin (14 mg) is dissolved in pH 7 0.1N phosphate buffer (2.5 ml) and the solution adjusted to pH using an automatic burette dispensing IN NaOH. While maintaining a pH of 8.5, methyl formimidate hydrochloride (25 mg) is added to the magnetically stirred solution. After 20 min. additional methyl formimidate (25 mg) is added. The reaction is allowed to proceed another 25 min and the pH is adjusted to 7.0 with 2.5NHC1. The entire solution is then chromatographed on Dowex 50-X4 resin (26 cc, Na+ cycle, 200-400 mesh) eluted with water. The N-methyl-N-formimidoyl derivative elutes in 3-4 column volumes and is lyophilized to a white Solid (3 mg).

UV (pH 7 0.1N phosphate buffer) max 298 nm IR ( Nujol mull) 1760, 1710 cm”1 NMR (100 MHz, D2O) J* 1.32 id, J=SHz, CH3 -CH) 3.12 (s, NCH3), 7,86 (s, N-£=N) EXAMPLE 2 N-Formimidoyl-N-ethyl thienamycin A solution of N-ethyl thienamycin (5.6 mg) in 2.8 ml of 0.2N pH 7 phosphate buffer is adjusted to pH 8.5 and ethyl formimidate hydrochloride (50 mg) is added. The pH is maintained at 8.5 by the addition of N sodium hydroxide sctetion under the control of a pH stat.

The mixture is stirred at 23’ for 25 minutes and another portion (50 mg) of formimidate is added. After 25 minutes the reaction mixture is neutralized with dilute hydrochloric acid and chromatographed on 1,5 x 15 cm column of Dowex 50-X4 + resin (Na cycle, 200-400 mesh). The column is eluted with water at a flow rate of 0.5 cc/min and the effluent is monitored by HPLC (1/4 x l'C18 Porasil; solvent 0.01N NaHPO, solution, flow rate - 2 cc/min; N-ethyl thienamycin retention time = 5 min, N-formimidoyl- N-ethyl thienamycin til * 9.5 min) The combined product fractions(UV 42 ODU at 298 mu) are evaporated to 5 ml and freeze dried to a white powder.

EXAMPLE 3 Silylation of N-methyl Thienamycin To a suspension of N-methyl thienamycin (8 mg) in 1 ml of tetrahydrofuran under a stream of nitrogen is added hexamethyldisilazane (0.1 ml) and trimethylchlorosilane (20 μΐ). The mixture is vigorously stirred at 25® for 20 minutes then centrifuged to remove ammonium chloride. The supernatant is evaporated to an oil under a stream of nitrogen and the residual oil is used directly in further reactions.

EXAMPLE 4 Cl© rOH N-Methyl-thienamvcin —-> Preparation of Thienamycin N-Methyl-N-piperidin-l-yl Methylene Derivative; N-Methyl-thienamycin (57 mg) is silylated according to the procedure previously described. The silylated antibiotic, is dissolved in methylene chloride (6 cc) in a septum stoppered flask under positive nitrogen pressure and cooled in a dry ice-acetone bath. To the magnetically stirred solution is added a solution (180 μΐ) of triethylamine (644 yrnol) in methylene chloride. This is followed by the addition of a solution of chloropiperidinomethylium chloride (67 mg., 405 μιηοΐ) in methylene chloride (465 ul). After 1 hourih a dry ice bath, the reaction solution is rapidly added to a tetrahydrofuran - pH 7.0, IN phosphate buffer (1:1) solution (50 ml). The mixture is then concentrated under vacuum to 10 ml to give a homogeneous solution.

The solution is washed twice with ethyl acetate (2x5 ml) S and ether (2 x 5 ml) and briefly pumped under vacuum.

This aqueous solution is then chromatographed on an XAD-2 resin column (60 ml bed). The product is eluted in 10% aqueous tetrahydrofuran (following water elution) to give the product which is recovered by lyophilization.

EXAMPLE 5 rOH Preparation of N-Methyl-N-benzimidovl Thienamycin N-Methyl-thienamycin (59 mg.) is dissolved in a 33% Ν,Ν-dimethylformamide pH 7 phosphate buffer (0.05N) solution (4.5 ml.) and adjusted to pH 9,5 using 2.5N NaOH with an automatic dispensing burette. The solution is magnetically stirred at 25*c. and methylbenzimidate-HCl (340 mg) is added at once. After 30 min. the solution is extracted twice with an equal volume of chloroform and adjusted with dilute aqueous phosphoric acid to pH 7.0.

The buffered solution is chromatographed on XAD-2 resin (65 ml.). The column is first eluted with water followed by 10% aqueous tetrahydrofuran which elutes the product. This fraction is concentrated to one-half volume and freeze-dried to give the product.

EXAMPLE 6Thi—OH ?h3 NC -NH, L-COjCHj sc= c /CH3 ./ CH, Br Preparation of N-Methyl N-benzimidoyl Thienamycin, 3-MethyI-2· Buten-l-yl Ester N-Methyl-N- benzimidoyl thienamycin (5.9 mg.) is dissolved in hexamethylphosphoramide (100 μΐ.) containing 1-bromo3-methyl-2-butene (4.8 μΐ,) and triethylamine (0.5 μΐ.) and magnetically stirred at 22°C. After 1 hour the crude reaction product is chromatographed on a 250, μ thick silica gel plate developed in 8:2, chloroform, ethanol. The band of R, 0.1 - R. 0.3 is removed and eluted with ethanol. XX,' N-methyl-benzimidoyl thienamycin, 3-methy1-2-buten-l-yl ester hydrobromide is isolated as a solid after precipitation from an ethanol-chloroform solution with hexane.

EXAMPLE 7 r-OH NH Th-j-NO; NH, h:o2h Preparation of N-Methyl-N-guanyl thienamycin N-methyl-thienamycin (8.9 mg) is dissolved in pH 7 0.1N phosphate buffer (0.7 ml) and Ν,Ν-dimethylformamide (0.3 ml) and the solution brought to pH 9.5 by the addition of 2.5N sodium hydroxide solution. To the magnetically stirred solution is added O-methylisourea-hydrogen sulfate (43 mg.) causing a slight drop in pH. Additional sodium hydroxide solution is added to bring the pH back to 9.5 •1·° and the solution is stirred 30 minutes at 23 *C. The solution is then neutralized to pH 7.0. The solution containing a mixture of N-methyl thienamycin and N-methylN-guanyl thienamycin is chromatographed on a 20cc of Dowex 50X4 resin (Na+ cycle, 200-400 mesh) and the product 15 recovered by lyophilization.

EXAMPLE 8 Silylated N-Methyl (CH3)2N»CHC1 Thienamycin -- Preparation of N-Methyl-N-Pimethylaminomethylene Thienamycin N-Methyl-thienamycin {16.5 mg) is silylated with hexamethyldisilazane (200 μΐ) and trimethylchlorosilane (60 μ,Ι) in the usual manner. The silylated N-methyl thienamvcin is suspended in '(ethanol free) chloroform (1 ml.) with magnetic stirring under a nitrogen atmosphere. The mixture is cooled to -45’C. and a solution of triethylamihe (21 μΐ) in chloroform (21μ1) is added followed by a solution of (chloromethylene)-dimethylammonium chloride (11.5 rag) in chloroform (50 μΐ). The mixture is warmed to -256c. during 1 hour and 0.1N pH 7 phosphate buffer (5 ml) is added. The mixture is vigorously stirred 15 minutes. The aqueous phase is separated and contains N-methyl-N-dimethylaminomethylene thienamycin which is recovered after chromatography on Dowex 50X4 resin.

EXAMPLE 9.

Th · r- OH + ^NH2 NC CH3n CO2BrCH2OCC(CH3)3 Th•OH CH3 •Ac + -NH,Br X; ;-o-ch2occ(ch3)3 Preparation of N-formimidoyi-N-Methyl-Thienamycin Pivaloyloxy methyl Estar Hydrobrcmide N-Formimidoyl-N-methyl thienamycin (10 mg.) is dissolved in hexamethylphosphoramide (200 μΐ.) containing bromomethyl pivalate (10 μΐ.) and triethylamine (1 μΐ.) and magnetically stirred at 22 °C. After 2 hours the hexamethylphosphoramide solution is dissolved in 2 mi. methylene chloride and the product precipitated with a 50% hexane-ether solution. The precipitate is dissolved in an aqueous 10% tetrahydrofuran solution and chromatographed on an Dowex XAD-2 resin packed column. N-Formimidoyl-N-methyl thienamycin pivaloyloxymethyl ester is isolated as a solid after tetrahydrofuran elution of the column, and lyophilization. 46S73 EXAMPLE 10 Preparation of N-Ethyl-N-trifluoroacetlmidoyl Thienamycin N-ethyl-thienamycin (199 mg) is dissolved in pK 7 0.1N phosphate buffer (7 ml) and adjusted to pH 8.5 with IN sodium hydroxide solution. While maintaining this pH with an automatic burette, a solution of methyl trifluoroacetimidate (355 μΐ) in dioxane (2.5 ml) is added at once. After 30 minutes the pH is readjusted to 7.0 by the addition of IN hydrochloric acid. The solution is then chromatographed on Dowex 50 X 4 resin (200 oc, Na+ cycle, 200-400 mesh) and is eluted with water. The N-ethylN-trifluoroacetimidoyl thienamycin derivative elutes in the first half column volume. This eluate is rechromatographed in a similar manner on Dowex 50 X 4 (100 cc.· Na+cycle, 200-400 mesh) and. the first column volume concentrated and chromatographed on Dowex XAD-2 resin (30 cc). The N-ethyl-N trifluoroacetimidoyl thienamycin derivative elutes in 2.5 5.0 column volumes which is lyophilized to a white sclid. 68 73 EXAMPLE 11 Preparation of N-Allyl-N-Acetimidoyl Thienamycin CH2-CH=CK2 CH,N-C=NH 2 ι , OH N-Allyl-thienamycin (190 mg) is dissolved in pH 7 0.1N phosphate buffer (13 ml) and cooled in an ice bath with magnetic stirring. The solution is adjusted to pH S.5 using 2.5N sodium hydroxide solution dispensed from an automatic burette. While maintaining a pH of S.5, ethyl acetimidate hydrochloride (400 mg) is added portionwise over a few minutes. After an additional 40 minutes the solution is adjusted to pH 7.0 with 2.5N hydrochloric acid. The solution is then chromatographed on Dowex 5Q-X3 resin (250 cc, Na+ cycle, 100-200 mesh) and is eluted with water. The N-allyl-N-acetimidoyl derivative elutes in 1-2 column volumes (240-520 cc) and is lyophilized to a white solid.

V EXAMPLE I2 Preparation of N-ethyl-M- [(4-pyridyl) (.imino)methyl)thienamycin N-Ethyl-thienamycin (80 mg) is dissolved in aqueous sodium bicarbonate (24.7 mg., 0.294 mmole in 2.0 ml) at 25®C. Methyl isonicotinimidate (80 mg., 0.588 mmole) is dissolved in the solution and progress of the reaction is followed by timed aliquots using high performance liquid chromatography (HPLC)t Waters instrument? 0.2 x 61 cm.

Cjg Bondapak reverse phase column; 1.5 ml/min aqueous 10% THF; UV (254 nm) and R.l. monitors. The reaction is essentially complete in 40 minutes, and the reaction solution is chromatographed directly over an 18.4 x 270 mm XAD resin column, first eluting with deionised, distilled water, then changing to aqueous 10% THF. The eluate is monitored by UV and HPLC is used tc locate the pure product. Correct fractions are combined and lyophilized to yield the product as a colorless, fluffy powder. 4687 EXAMPLE 13 Following the procedure of Example 12 , but replacing the reagent with an equivalent amount of methyl 4thiazolecarboximidate, there is obtained;· N-ethyl-N[(4-thiazolyl)(imino)methyl] thienamycin.

EXAMPLE 14 Preparation of N-Allylformaraide A mixture of allylamine (5..00 g., 87.6 mmole) and methylformate (5.26 g., 87.6 mmole) is stirred at ’C., for 2 hours. At the end of this time, the reaction flask is fitted with a short path distillation head and the desired N-allylformamide is collected at 89-90’C./0.7 mm as a colorless oil. Yield 7.0 g. (94%). XR(CHCl3) 3380, 1680 cm nmr (CHClj) EXAMPLE 15..

Preparation of Ethyl N-Allvlformimidate-Hvdrochloride Ethyl chloroformate (2.66 g., 24.47 mmole) is added by syringe to N-allylformamide (2.08 g., 24.47 mmole) in a dry flask under N2. The resulting mixture is then stirred at 25*C., for 2 hours during which time C02 ·*·Ε rapidly evolved. The reaction mixture is then heated to 45°C. until no further evolution of gas is evident (2 hours) The viscous product is then cooled and held at a vacuum of 0.2 mm for 2 hours to remove all volatiles.

EXAMPLE 1 g Breparation of Methyl N-Dimethylaminoformimidate To a stirred solution of Ν,Ν-dimethylformhydra2ide (0,22 g) in 2.0 ml of chloroform, under nitrogen, is added methyl chloroformate (0.5 ml). The mixture is heated at 40*C. for three hours then evaporated under nitrogen.

The mixture is triturated with anhydrous ether. The supernatant solution is decanted and the residue dried in a stream of nitrogen.

Yield: 284 mg. nmr CDC^ei, 9.13 (CH) j 3.80 (OCH^) , 3.01(N(CH3)2).

EXAMPLE 17 Preparation of N-Cyclopropyl Formamide A mixture of cyelopropylamine (5.00 g, 87.6 mmole) and methylformate (5.26 g, 87.6 mmole) is stirred at 25®C., for 2 hours, (an initial exotherm is noted).

The mixture is then placed on the rotary evaporator to remove the MeOH formed in the reaction. The remaining material is distilled through a short path head to yield 2® 6.92 g (93%) of the desired N-cyclopropyl formamide as a colorless oil, n.m.r. (CLCi3) £/‘3.1 (IH, br S), 6.S-8.5 (IH, br), /'2.4-3.0 (lH, m), £ 0.4-1.0 (4H,m). - 6δ 4 6 8 7 3 EXAMPLE ig Preparation of Ethyl N-Cyclopropyl Formimidate Ethyl ohloroformate (4.078 g, 37.58 mmole) is added by syringe to N-cyclopropylformamide (3.194 g, 37.58 mmole) in a dry flask under N2· After an induction period of 30 sec., a rapid evolution of gas begins. The resulting reaction mixture is stirred at 25*C until no further evolution of gas can be detected («. 4 hr), then the viscous product is subjected to a vacuum of 0.5 mm for 1 hr to remove any unreacted ethyl ohloroformate. NMR analysis of the product shows the formyl proton at 9.37 as a broad singlet. (CDCl^ solution).

EXAMPLE 19 Preparation of N-Ethyl-N'-Dimethylamino-N-Formimidoyl Thienamvcxn r OH zch3 >0H3 Th N-N. c2hs I- COOH N-Ethyl-thienamycin (115 mg) is dissolved in pH 7 0.1N phosphate buffer (7 ml) and the pH of the solution is adjusted to 8.5 using an automatic burette dispensing IN NaOH. To this stirred solution is added methyl N-dimethylaminoformimidate hydrochloride (284 mg) while the pH is maintained at 8.5. After 20 minutes the pH of the solution is adjusted to 7.0 using 2.5 N HC1 and the solution is J, chromatographed on Dowex 50-X4 resin (53 cc, Na cycle, 200-400 mesh) eluted with deionised water. The chromtocraphy is carried out in a water jacketed column at 3*. The N-Ethyl Ν'-dimethylamino-N-formimidoyl derivative elutes in 2 column volumes and is lyophilized to a white solid.

EXAMPLE 20 Preparation of N,N'-Dimethyl-N-Formimidoyl Thienamycin •OH Th A'? ch3 *- COOH N-Methyl thienamycin (140 mg) is dissolved .in pH 7 0.1N phosphate buffer (10 ml) and the pH of the solution is adjusted to 8.5 using an automatic burette dispensing IN NaOH. To this solution is added methyl N-methyl formimidate hydrochloride (200'μΐ) while the pH is maintained at 8.5. After 40 minutes the pH is adjusted to 7.0 using 2.5N KC1 and the solution is chromatographed on Dowex 50-X4 resin ‘‘θ (72 cc, Na+ cycle, 200-400 mesh) eluted with deionized water. The Ν,Ν'-dimethyl-N-formimidovl derivative elutes in 2 column volumes and is lyophilized to a white solid. - 70 46873 EXAMPLE 21 Preparation of N-allyl - Ν'-Benzyl-N-Formimidoyl Thienamycin Th--0H NCH,0 II .NO Ph ch2-ch«ch2 Lcooh N-Allyl-thienamycin (110 mg.) is dissolved in pH 7 O,1N 5 phsophate buffer (7 ml.) and the pH of the solution is adjusted to 8.5 using an automatic burette dispensing IN NaOH. A solution of ethyl N-benzyl formimidate flucroborace (572 mg) in ja-dioxane (2 ml) is added to the buffered solution while the pH is maintained at 8.5. After 20 minutes the pH of the solution is adjusted to 7.0 using 2.5 N HCI and chromatographed on Dowex 50-X4 resin (53 cc, Na+ cycle, 200-400 mesh) eluted with deionized water. The chromatography is carried out in a water jacketed column at 3. The N-allyl-Ν'-benzyl-Nformimidoyl derivative elutes in 2 column volumes and is lyophilized to a white solid.

EXAMPLE 22 Preparation of N-ethyl-N1-isopropyl-N-formimldoyl thienamycin Thr-OH NC· nch(ch3)2 C2H5 J—COOH N-ethyl-thienamycin (110 mg) is dissolved in pH 7 0.1M phosphate buffer (7 ml.) and the pH of the solution is adjusted to 8.5 using an automatic burette dispensing 1 N NaOH.

A solution of methyl N-isopropyl formimidate hydrochloride (300 mg.) in p-dioxane (1 ml.) is added to the magnetically stirred buffered solution while the pH is maintained at 8.5. After 25 minutes the pH of the solution-is adjusted to 7.Q using 2.5 N NaOH and chromatographed on Dowex 50-X4 resin (53 cc, Na+ cycle, 200-400 mesh) eluted with deionized water.

The chromatography is carried out in a water jacketed column at 3°C. The N-ethyl-N'-isopropyl-N-formimidoyl derivative elutes in 2 column volumes and is lyophilized to a white solid. - 72 46873 EXAMPLE 23 Preparation of N-Methyl-M-CW'-Allyl-Formiraidoyl) Thienamycin rOH ,/ch2ch-ch2 Th-f-N-Cx.

CH3 *<00H To a prechilled sample of thienamycin (123 mg., 0.452 mmols) is added 13 ml. of cold 0.1N phosphate buffer.

The solution is adjusted to pH 9 with N sodium hydroxide.

To this basic solution at 2°C. is added all at once ethyl N-allyl-formimidate hydrochloride (0.3 g).

The pH drops to 7.3 and is brought back to S.5 with additional sodium hydroxide. The reaction mixture is stirred at 2*C for an additional 30 min.and the pH is adjusted to 7 with cold 0.1N sulfuric acid. The reaction mixture is chromatographed on a Dowex-50 X 4 column (60 ml., 200-40C mesh) eluting with water at a flow rate of 0.5 ml/cia of resin bed. After discarding the first 400 ml of eluate, the next 150 ml. is lyophilised to give the product.

EXAMPLE 24 Preparation of N-Methyl-N(3-azidopropionimidoyl)Thienamycin -OH Th--NCCH-CH.N, ! 2 2 3 CH, *-C00H To a solution of N-methyl-thienamycin (133 mg) in 10 ml . 0.1M pH .7.0 phosphate buffer is added 1.2 g of O-ethyl-3-azidopropionimidate’HCl while the solution is maintained at pH 8.5 with 2.5 N NauE. The mixture is stirred at 0°C for 0.5 hr., then is neutralized with 2.5N HCl to pH 7.0, concentrated to 5 ml. and chromatographed on a Dowex 50W X 8 (Na form) column (1.5M x 12) which is eluted with water to give the desired product.

EXAMPLE 25 Preparation of N-Methyl-N(3-aminopropionimidoyl)thienamycin Th OH - -N^CH2CH?N. CH3 -CO^a® H2/Pd rOH Th-. «Π © NCCH,CH-NK, I 2 2 3 CH, Lco2© (i) (II) - 74 46873 EXAMPLE 26 Preparation of N-Methyl-N-Nitroguanyl Thienamycin N-Methyl-thienamycin (131 mg) is dissolved in a solution of dimethyl sulfoxide ( 10 ml), tri-n-butylamine (0.30 ml), and 2-methyl-l-nitro-2-thiopseudourea (0.3g). The solution is heated in a water bath at 45’C.while a stream of nitrogen is vigorously bubbled into the solution.

After 50 min. the solution is concentrated under high vacuum to 1.0ml. and dissolved in 0.C5N pK7 phosphate « buffer (7 ml). The unreacted thiopseudourea is precipitated and removed by filtration. The solution is then chromatographed on Dowex 50-X4 resin (53 cm3, 200 400 mesh, Na+ cycle) and eluted with water. The N-nitroguanyl derivative elutes in the first column volume and is lyophilized to a solid .

EXAMPLE 27 Preparation of Ethyl N-Bensylformlmidate A solution of 690 mg (5.1 mmoles) of N-benzylformamide in 5 ml of methylene chloride is cooled in an ice-water bath and put under an argon blanket. Tha solution is stirred while 4.9 ml (4.9 mmoles) of 1M triethyloxonium flucrofccrate in methylene chloride is added dropwise. After a 45 minute reaction time, the mixture is concentrated to dryness under reduced pressure at room temperature, and the residue is dried under reduced pressure over P,O.. The nuclear magnetic resonance spectrum of the product in deuterochloroform is' fully in accord with the product being a fluoroborate etherate complex of ethyl N-benzylformimidate.

EXAMPLE 28 Preparation of N—isopropyl formamide Formartide (1.13 g, 0.98 ml) is dissolved in 10 ml of toluene, containing toluenesulfonic acid (4.7 g). To tl)e above mixture is added isopropylamine (2.-95 g,,4.25 ml). The mixture is refluxed overnight ί under a gentle stream of N^· The solution is filtered and the toluene is evaporated under reduced pressure.

The residual oil is· distilled at 59-62°C/.O7 mm to 10 give 1.0 g of the desired product.

EXAMPLE rg Ffaparation cf Methyl Ν-isopropyl formimidate Isopropyl formamide (535 mg) is treated with an equivalent-amount of ethyl chloroformate (44 0 μΐ) f0r 2-3 hours under N., at 40-45°C. The mixture is washed successively with petroleum ether anhydrous ether and benzene leaving the product as an oil. 4S873 EXAMPLE 30 Preparation of N-ethyl-N-[N'-cyclopropylformimidoyl] thienamycin rOH ThN-C.

*H Aooh N-Ethyl-thienamycin (100 tg) in 10 ml 0,1 M, pH7.0 phosphate buffer is adjusted and maintained at pH 8.5 - 9.0 while 300 mg of ethyl N-cyclopropylformimidate hydrochloride is added dropwise to the solution. The mixture is stirred at 23· for 40 minutes, then is neutralized, and chroma10 tographed on a Dowex - 50 X 8 (Na form) ion-exchange column (1.5 x 10). The column is eluted with water, collecting 6.5 ml fractions. Fractions 43-95 are combined, concentrated and freeze-dried to give the solid product.

V EXAMPLE 31 Preparation of 0, N-Dimethyl-N-(p-Nitrobenzyloxycarbonyl)Thienamycin-p-Nitrobenzyl Ester Th. -och3 -^3-0-<*2-ΌNO, COOCH, NO, Step A N-Methyl-N-(p-Nitrobenzyloxycarbonyl·)thienamycin Lithium Salt To N-methyl thienamycin (220 mg. in 60 ml. water at 0°C) is added successively, 679 mg. NaHCO^, 60 ml dioxane and then with stirring 1.1 equivalents p-nitrobenzyl chloroίθ formate 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, 50V/cm., 20 minutes) shows no free Nmethyl thienamycin present. The aq. extract is adjusted to pH 2.2 with 1M H^PO^ solution and extracted three times with EtOAc. The EtOAc extract is dried over MgSO4, filtered and reextracted with 0.1N LiOH, to pH 8.2. The final pH is adjusted to 7.0 with 1M H^PO,. and the sample lyophilized.

Step Β N-Methyl-N-(p-Nitrobenzyloxycarbonyl)Thienamycin(pnitrobenzyl)ester A mixture of p-nitrobenzyloxycarbcnyl-N-methyl-thienamycin lithium salt (295 mg) and 0.4g of p-nitrobenzyl bromide in 3 ml of hexamethyl phosphoramide is stirred for 3 hours at 25*C. The solution is diluted with 50 ml of ethyl acetate and extracted successively with water (3 portions), pH 7 phosphate buffer and saturated sodium chloride solution. The organic phase is dried over magnesium sulfate and evaporated to 5 ml causing the product to crystallise.

Tr.e crystals are collected and washed with ethyl acetate.

Step C 0,N-Dimethyi-N-(p-Nitrobenzyloxycasbonyl)-thienamycin(pNitrobenzyi Ester To a solution of 135 mg. of N-methyl-N-p-nitrobenzyloxycarbonyl thienamycin (p-nitrobenzyl) ester in 50 ml. of methylene chloride at 0®C. is added with vigorous stirring 0.5 ml of 0.006 M fluoroboric acid in ether-methylene chloride (3:1) immediately followed by 10 ml of a cooled solution of 0.6M diazometnane in methyl chloride. The diazomethane is decolorized in one minute. The solution is extracted with 10 ml of 0.1N phosphate buffer, dried and evaporated to a small volume. The solution is applied to two 8x8 1000p silica gel plates which are developed with 3:1 ethylacetate-chloroform. The band containing Ο,Ν-dimethyl N-p-nitrobenzyloxycarbonyl thienamycin p-nitrobenzylester is eluted with ethyl acetate and the product is recovered by evaporation of the eluate. 4«873 J . EXAMPLE 32 0, N-Dimethyl Thienamycin A solution of 20 mg. of 0, N-Dimethyl N-(pnitrobenzyloxycarbonyl)thienamycin p-nitrobenzyl ester in 2 ml of tetrahydrofuran and 1 ml of ethanol is hydrogenated at 50 psig, 23’C in the presence of 20 mg of platinum oxide for 2 1/2 hours. The catalyst is filtered and 1 ml of 0.1N 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 cf water and 5 ml cf 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 aid then with 10% tetrahydrofuran. The tetrahydrofuran eluate is concentrated and lyophilized giving substantially pure 0, N-dimethyl thienamycin. - 80 468*73 EXAMPLE 33 Preparation of 0, N-Dimethyl-N-Formimido^i Thienamycin eZ^/BCH ”O t NH CH. '3 co2h 0, N-Dimethyl thienamycin (14 mg) is dissolved in pH 7.0 5 IN phosphate buffer (2.5 ml) and the solution adjusted to pH 8.5 using an automatic burette dispensing IN NaOH.

While maintaining a pH of 8.5, methyl formimidate hydrochloride (25 mg) is added to the magnetically stirred solution. After 20 min. additional methyl formimidate (25 mg) is added. The reaction is allowed to proceed anothe 25 min and the pH is adjusted to 7.0 with 2.5NHC1. The entire solution is then chromatographed or. Dowex 50-X4 resin (26 cc, Na+ cycle, 200-400 mesh) eluted with water. The 0,N-aimethyl-N-formimidovl derivative eluates in 3-4 column volumes and is recovered by lyophilisation.

V EXAMPLE 34 Preparation of O-Acetyl-N-Methyl-N-(p-nitrobenzyloxycarbonyl)-thienamycin (p-nitrobenzyl)ester To a solution of 50 mg of N-methyl-N-(p-nitrobenzyloxycarbonyl) thienamycin g-nitrobenzyl ester in 0.5 ml of pyridine is added 0.16 ml of acetic anhydride. The mixture is allowed to react at 25°C., for threshours, then pumped to dryness under vacuum. The solid residue is dissolved in chloroform and chromatographed on an 8x8 ΙΟΟΟμ silica gel plate in 3:1 ethylacetatechloroform yielding o-acetyl-N-methyl-N-p-nitrobenzyloxy carbonyl thienamycin p-nitrobenzyl ester.

! ( - . EXAMPLE 35 O-Acetyl-N-methyl thienamycin A solution of 20 mg. of O-acetyl-N-methyl-N-(pnitrobenzyloxycarbonyl)thienamycin ρ-nitrobenzyl ester in 2 ml of tetrahydrofuran and 1 ml of ethanol is hydrogenated at 50 psig, 23’C in the presence of 20 mg of platinum oxide for 2 1/2 hours. The catalyst is filtered and 1 ml of 0.1N pH 7 phosphate buffer is added to the filtrate.

The solution is evaporated under reduced pressure to 2 ml 2·° 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 ani then with 10% tetrahydrofuran. The tetrahydrofuran elvate is concentrated and lyophilized giving substantially pure 0-acetyl-N-methyl thienamycin. >46873 EXAMPLE 36 Preparation of O-Acetvl-N-Methyl-N-Formimidovl· thienamycin OCOCH3 C02h 0-Acetyl-N-methyl thienamycin (14 mg) is dissolved in pH 7 0.1N phosphate buffer (2.5 ml) and the solution adjusted to pH 8.5 using an automatic burette dispensing IN NaOH.

While maintaining a pH of 8.5, methyl formimidate hydrochloride (25 mg) is added to the magnetically stirred solution. After 20 min. additional methyl formimidate (25 mg) is added. The reaction is allowed to proceed anothe min and the pH is adjusted to 7.0 with 2.5NHC1. Tha entire solution is then chromatographed on Dowex 50-X4 resin (26 cc, Na+ cycle, 200-400 mesh) eluted with water. The fractions containing O-acetyl-N-methyl-N15 formimidoyl thienamycin are combined and lyophilized.

EXAMPLE 37 Following the procedure set forth in the foregoing text and examples, the following compounds of the present invention are obtained. The reagents, imido ethers and imido halides, utilized in the reaction with thienamycin, or a derivative thereof, to provide the following compounds are either known, or may be prepared as described above. 1.) CHg ch3 H H 2.) CH3 CK3 ch3 CH3 3.) <*3 CH3 ch3 H 4.) CH3 N(CIi3)2 H H 5.) CH3CF3 H H 6.) ch3 o H H 7.) ch3 G jj tl 8.) ch3 G H H 9.) ce3 H H 10.) ch, H -ch2ch2-s-ch3 H 11.) Ch-3 H -CH(CH2)3 H 12.) CH3 H -ch(ch3)3 H Com- pound R R6' R1 R2 13.) ch-3 H -CH2CH3 B 14.) CH- 3 . H -CH2-CH«=CH2 H 15.)Cii3 H -CH2-C=»CH2 CHj H 16.) ch3 H -< H 17.)CH3 CH2CH3 H H 18.) ch3 B -ch2cf3 H 19.) ch3 H -CH2C00Na H 20.). ch3 H -och3 H 21.) ch3 H ch2ch=ch-ch3 H 22.) ch3. B H 23.) ch3 - H . -o B 24.) ch3 B Ό H 25.) ch3 . H Ό B 26.)CIi3 H O B 27.) ch3 B Ό H 23.) ch3 HCH2“O JJ 29.) ch3 H -^-och3 H 30.) CH- ύ B H 31.) CH3 a · ΓΛ H 32.) ch3 B H 33.) CK3 B -CH2CH2\_y° H 34.) ch3 H -ch2ch2-0 H Com- pound R R6 1 R2 35.) ch3 H H 36.) ch3 Η -ch2-O-sch3 H 37.) ch3 H -CH2-O’N(CH3)2 H 38.) ch3 H ch3C2H5 39.) ch3 HC2H5C2H5 40.)CH3 H CHj ck(ch3)2 41.) ch3 H ch3 0 42.) ch3 H ch3 CH2CH*CH2 43.) ch3 ~OCH- z H H 44.) 'ch3 Η H 45.) ck3 “C/° H H 46.) ch3 -_jr^-cH3 H H 47.5 48.) CH, 0 CH3 -Q _rvCK3 H H H H 49. ) 50. ) ch3 ch3 -ch2J^z H H H 51.) ch3 CH3 “C2H5 H 52.) ch3 CK3 -CH(CH3)2 H Com- pound R s6 R1 R2 53.) ch3 CH3 -ch3 ~C2H5 54.) CH. j ' CH3 -cwh3>3 H 55.) Η -CH-CSCH H 56.) -C2H5 ch3 Η H 57.) -c2h5 H -ch3 H 58.) -C2H5 K -ch3 ch3 59.) -C2H5 ch3 -ch3 60.) -ch20 H H H 61.) -CH20 H -ch3 H 62.) -ch3 • H -ch2ch2oh H 63.)CH3 H -CH2CK2N(CK3)2 H 64.) ch3 H -CH,CH2N(CH3)2 H 65.) ch3 nh2 ch3 H 66.) ch3 nh2 ch3 ch3 67.) ch3 nh-ch3 H H 68.) CH, NHCH, ch3 H 69.) ch3 nh2 nh2 H 70.) ch3 sch3 H H 71.)CH3 sch3 ch3 H 72.) ch3 H och3 H 73.) CK3 -CH2Br H H 74.) CK3 -ch2n3)2 H H Ο .0 Com- pound R R' Y A© RH X 75.) ch3 -CH2CH«C(CH3)2 H NH2 H Cl 76.) ch3 -ch20-°ch3 H NH2 H Cl 77.) CK3 -CH2OC-C(CH3)3 0 H NHCH3 H Cl 78.) ch3 -CH2OCC(CH3)3 0 H NH2 ch3 CH3COO 79.) ( 2H5 -CH2O-C-C(CH3)3 0 H Cl 80.) CK3 -ch2ch2 -s-ch3 H NH2 ch3 H2P°4 81.) ch3 -CH2-O-C-CH3 0 H NHCH3 H Cl 82.) ch3 -CK--C-0 i H 0 H NH2 H Cl 83.) CK3 5-indanyl H NH2 H Cl 84.) ch3 phthalidyl H NH2 H Cl 85.) ch3 Na © S03 NH2 H - 86.) ch3 Na P03H® NH, H - 87.) ch3 -CH2OC-C(CH3)3 0 nh2 H - 88.) ch3 -CH2O-C-C(CH3)3 po3h© nh2 H - ο EXAMPLE 38 Preparation of Pharmaceutical Compositions One unit dosage form is prepared by mixing 120 mg. of N-Methyl—N-Formimidoyl thienamycin 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 using 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., compressed tablets and pills can also be prepared. The following examples are illustrative of the preparation of pharmaceutical formulations: TAELET N-Methyl— N-Formimidoyl thienamycin Cornstarch, U.S.P.

Dicalcium Phosphate Lactose, U.S.P.

PER TABLET 125 mg. mg. 192 mg. 190 mg.

The active ingredient is blended with the dicalcium 20 phosphate, lactose and about half of the cornstarch. The mixture is then granulated with 15% cornstarch paste (6 mg) and rough-screened. It is dried at 45*C and screened again through No 16 Screens (US Standard mesh). The balance of the cornstarch and the magnesium scearate is added and the mixture is compressed into tablets, approximately 0.5 inch in diameter each weighing 800 mg.

PARENTERAL SOLUTION Ampoule; N-Methyl—N-Formiraidoyl thienamycin 500 mg. 4 6 8 7 3 CPTHALMIC SOLUTION N-Methyl—N-Formimidoyl thienamycin yoo mg. Hydroxypropylmethyl 5 mg. Sterile Water · to 1 ml.

OTIC SOLUTION N-Methyl-N-Formimidoyl thienamycin Ιθθ m9· Eenzylalkylammonium chloride 0.1 mg. Sterile water to 1 ml.

TOPICAL OINTMENT IO N-Methyl-N-Formimidoyl thienamycin Polyethylene Glycol 4000 U.S.P. Polyethylene Glycol 400 U.S.P. 100 mg. 400 mg. 1.0 gram The active ingredient in the above formulations may be administered alone or in combination with other biologically active ingredients, 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 (5)

CLAIM S;1. A compound having the structure in which each of R1 and R2, independently of the other, 5 is a radical of the type defined for R, a hydrogen atom, or a nitro,' hydroxyl, C^_g alkoxyl, amino, C^_g alkylamino, di.(C^_g alkyl) amino or tri(C1_g alkylamino) radical, an extra anion being present in the latter case; or R and R are joined together to form, together with 10 the nitrogen atom to which they are attached, a substituted or unsubstituted monocyclic or bicyclic heteroaryl or heterocyclyl residue containing 4-10 ring atoms, one or more of which may be an additional hetero atom selected fran oxygen, sulphur and nitrogen; R is a cyano group or a sub15 stituted or unsubstituted carbamoyl, carboxyl, alkoxy)carbonyl, C1_^o alkyl, alkenyl, C2_lo alkynyl, c3_j_q cycloalkyl, C4_12 cycloalkylalkyl, Cg_12 cycloalkylalkenyl, c3_]_q cycloalkenyl, Cg_^2 cycloalkenylalkenyl, C4_i2 cycloalkenylalkyl, cg_^0 aryl, Cy_^g 20 aralkyl, Cg_^g aralkenyl, Cg_^g aralkynyl or monocyclic or bicyclic heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalkyl comprising 4 to 10 ring atoms one or more of which is a hetero atom selected from oxygen, sulphur and nitrogen and in which the alkyl residue of 25 the heteroaralkyl or heterocyclylalkyl radical contains from 1 to 6 carbon atoms; the substituent or substituents on R, R , Rz or on the ring formed by joining Rx and R are chlorine; bromine; iodine; fluorine; azido; alkyl; mercapto; sulpho; phosphono; cyanothio (-SCN); 5 nitro; cyano; amino; hydr azino; amino or hydrazino having up to three C-^g alkyl substituents; hydroxy; C^_g alkoxy; C^_g alkylthio«carboxyl; oxo; (C^_g alkoxy)carbonyl; 02_^θ acyloxy; carbamoyl; (C1_4 alkyl)carbamoyl or di(C^_4 alkyl)carbamoyl; R is a hydrogen atom, an acyl radical 4 4 10 or a radical of the tyne defined for R ; R is cj__3_0 j alkyl; substituted methyl carbonyl (c^_g alkoxy)-(C^_g alkyl); (C3_g cycloalkoxy)-{C,_g alkyl); C2_12 alkanoyloxyalkyl; partially or completely halogenated C^_g alkyl in which the halogen(s) is/are chlorine, bromine or 15 fluorine;aminoalkyl; 02_^θ alkenyl; C2_^Q alkynyl; acyl; ^3-14 a&oHycaib'WiaWl; C 21dialkylaminoacetoxyalkyl; C2_13 alkanoylaminoalkyl; ar-(C^_3 alkyl) in which the aryl residue contains from 6 to 10 carbon atoms; monocyclic or bicyclic heteroaralkyl or heterocyclylalkyl containing 20 4 to 10 ring atoms, 1 to 3 carbon atoms in the alkyl residue, and 1 to 4 hetero atoms selected from oxygen, sulphur and/or nitrogen; nuclear-substituted aralkyl or heteroaralkyl in which the substituent is chlorine, fluorine, bromine, iodine or C^_g alkyl; aryl or nuclear25 substituted aryl containing 6 to 10 ring carbon atoms and in which any nuclear substituent is hydroxy, 01-6 alkyl, chlorine, fluorine or bromine, aralkoxyalkyl; C2_^2 alkylthioalkyl; C4_12 cycloalkylthioalkyl, (C2_1Q acylthio)-(C3_g alkyl); or phenylalkenyl in which alkenyl 30 has 2-6 carbon atoms; R8 is substituted or unsubstituted alkyl; Cf-lQ alhenyl or alkynyl; ring substituted and unsubstituted cycloalkyl, cycloalkenyl, cycloalkenylalkyl, and cyeloalkvlalkyl having 3-6 ring carbon atoms and up to 6 carbon atoms in any chain; 6-10 aryl; aralkyl having 6-10 ring carbon atoms and 1-6 carbon atoms in the alkyl chain; monocyclic or bicyclic heteroaryl or heteroaralkyl containing 4-10 ring atoms, one or more of which is oxygen, nitrogen or sulphur, 5 and 1-6 carbon atoms in the alkyl chain; and the ring or chain substituent(s) is/are chlorine, bromine, iodine, fluorine, azido, cyano, amino, C^g alkylamino, di(C^_g alkyl)amino or tri(C, . alkylamino) radical, an extra anion being present in the latter case, hydroxy, C^_g 10 alkoxy, G3_g alkylthioalkyl; carboxyl; oxo, (C^_g alkoxy)carbonyl; C2-10 acY1OxYf carbamoyl; (Ct_4 alkyl)carbamoyl; di(C^_4 alkyl)carbamoyl; cyanothio (-SCN) or nitro; R is 1 2 hydrogen, hydroxy, mercapto, R, -OR, -SR or NR R , where

1. 2 with an amine of formula NHR R in which X' is an -OR or -SR leaving group, R being as defined in Claim 1, R 1 , 1 2 in which R and R are as defined in Claim 1, each X' is 5 a halogen atom, X° is oxygen or sulfur and -X°R* 1 is a leaving group. 30. A process for preparing a compound according to Claim 3 comprising treating a compound of formula: 1 to 11, in which R is selected from the group consisting of: hydrogen, formyl, acetyl, propionyl, butyryl, chloroacetyl, methoxyacetyl, aminoacetyl, methoxycarbonyl 5 ethoxycarbonyl, methylcarbamoyl, ethylcarbamoyl, phenylthiocarbonyl, 3-aminopropionyl, 4-aminobutyryl, N-methylaminoacetyl, Ν,Ν-dimethylaminoacetyl, an Ν,Ν,Ν-trimethylaminoacetyl salt, 3-(Ν,Ν-dimethyl)aminopropionyl, a 3(Ν,Ν,Ν-trimethyl)amino propionyl salt, an Ν,Ν,Ν-triethyl10 aminoacetyl salt, a pyridiniumacetyl salt, guanylthio3 acetyl, guanidinoacetyl, 3-guanidinopropionyl, N -methylguanidinopropionyl, hydroxyacetyl, 3-hydroxypropionyl, acryloyl, propynoyl, malonyl, phenoxycarbonyl, amidinoacetyl, acetamidinoacetyl, amidinopropionyl, acetamidi15 nopropionyl, guanylureidoacetyl, guanylcarbamoyl, carboxy methylaminoacetyl, sulfoacetylaminoacetyl, phosphonoacety 3 laminoacetyl, N -dimethylaminoacetamidinopropionyl, ureidocarbonyl, dimethylaminoguanylthioacetyl, 3-(1methyl-4-pyridinium)propionyl, 3-(5-aminoimidazol-l-yl)20 propionyl, a 3-methyl-l-imidazoliumacetyl salt, 3-sydnonylacetyl, o-aminomethylbenzoyl, o-aminobenzoyl, -p(och 3 ) 2 . S II / 0Cf! ‘OM s ΌΜ -p/n(ch 3 )27 2 101 OM -P-N(CH,),, \ 3 2 OM -S-OM, OM where M is hydrogen or cation. an alkali or alkaline-earth metal 28. A compound as claimed in any one of claims 1-11 and 27 in which 5 X is OR 4 , where R 4 Is hydrogen, methyl, t-butyl, phenacyl, £-bromophenacyl; pivaloyloxymethyl, 2,2,2-trichloroethyl, allyl, 3-methyl-2-butenyl, 2-methyl-2-propenyl, benzyl, -j benzhydryl, p-t-butylbenzyl, phthalidyl, phenyl, 5indanyl, acetylthiomethyl, acetoxymethyl, propionyloxy10 methyl, methallyl, 3-butenyl, 4-pentenyl, 2-butenyl, aceto( xyacetyImethyl, pivaloylacetyImethyl, ' dimethylaminoethyl, methoxymethyl, S-indanylmethyl, benzyloxymethyl, methylthioethyl, dimethylaminoacetoxymethyl, acetamidomethyl, acetylthio15 ethyl, pivaloyIthiomethyl or methylthioinethyl. 29. A process for preparing a compound according to Claim 2 comprising treating a compound of formula: 5 3 in which R is Cj_g alkyl and R and X are as defined in 20 Claim 1 with an imido ester of formula: 102 1-2 R, R and R are as defined above; 15 X is hydroxy, mercapto, amino,acyloxy, -OR , -SR 4 , -NHR 4 , -N-R 4 , -0M, -OQ or, when the compound is In R 4 zwitterionic form, -0 , in which case A~ is absent; A, when the compound is not zwitterionic form, is a counter ion; 20 ; M is a pharmaceutically acceptable cation; and Q is a blocking group as herein defined. £ 2. 3 5 R and R are also as defined in Claim 1 and R is a 5 c i_6 alkyl group. 32. A process for preparing a compound according to Claim 4 comprising treating a compound of formula: j with an alkylating agent or analogous R-containing agent i 10 calculated to provide a compound of formula: 12 3 in which A,R,R ,R ,R and X are as defined in Claim 1, r5 is a C^_g alkyl group and X° is oxygen or sulfur. 33. A pharmaceutical composition comprising a 15 compound according to Claim 1 and a pharmaceutical 104 carrier therefor. 34. A pharmaceutical composition comprising, in unitary dosage form, a therapeutically effective amount of a compound according to Claim 1 and a pharmaceutical 5 carrier therefor. 35. A composition as claimed in Claim 33 or 34, in which the compound is a compound as claimed in any one of Claims 2 to 28, 36. A composition as claimed in any one of 10 Claims 33 to 35 that also contains an antibiotic and/or therapeutic agent that is not a compound as claimed in Claim 1. 37. A composition as claimed in any one of Claims 33 to 36 in orally ι administrable form, 15 38. A composition as claimed in any ope of Claims 33 to 36 in injectable form. 39. A composition as claimed in any one of Claims 33 to 36 in topical form. 40. A composition as claimed in any one of 20 Claims 33 to 36 in the form of a suppository. 41. A composition as claimed in Claim 37 in the form of a tablet or capsule. 42. A composition as claimed in Claim 38 in the form of an ampoule. 105 43. A composition as claimed in Claim 39 in the form of an ointment or an ophthalmic or otic liquid. 44. A composition as claimed in Claim 37 in the form of an aqueous or oily suspension, solution, emulsion, 5 syrup, elixir, dry powder for reconstitution with liquid , or a pill. 45. A composition as claimed in Claim 38 in the form of an aqueous or oily suspension, solution, emulsion, or a dry powder for reconstitution with sterile liquid. 10 46. A composition as claimed in Claim 39 in the for® of a cream, lotion, paint, powder, semisolid, ophthalmic or otic capsule or drops. 47. A composition as claimed in any one of Claims 33 to 36 in the form of a powder or liquid spray or 15 inhalant, a throat paint or a lozenge. 48. A composition as claimed in any one of Claims 33 to 36 in the form of an intramammary preparation for veterinary use. 49. A disinfectant containing as antibacterial 20 ingredient a compound as claimed in any one of Claims 1 to 28. 50. An animal feed or human food containing as antibacterial ingredient a compound as claimed in any one of Claims 1 to 28. 51. Water-based paint or paper-mill white water containing as antibacterial ingredient a compound as 106 claimed in any one of Claims 1 to 28. 52. A composition as claimed in Claim 33, substantially as hereinbefore defined with reference to Example 38.

2. A compound according to Claim 1, in which R is hydrogen or R, where R is as defined in Claim 1, and R® is C^_g alkyl. 25 3. A compound according to Claim 1, in which R® is -NR 1 R 2 and R 5 is C^g alkyl.

3. 5 10 in which R and X are as defined in Claim 1 and R is Cj_g alkyl with a compound of formula: X°R‘ ' r 1 r 2 n-c=n-r 1 in which R 1 and R 2 are as defined in Claim 1 and X° and -X°R'' are as defined in Claim 29. 15 31. A process for preparing a compound according to Claim 3 comprising treating a compound of formula: 103 4. 6 8 7 3 NR 1 R-C“X°R'’ or an imido halide of formula: R 1 R 2 N®=C-X' ,θ

4. A compound according to Claim 1, in which R® is hydroxy, mercapto, -OR or -SR, where R is as defined in Claim 1, and R® is C^_g alkyl. 30 5. A compound according to Claim 2, in which each of R 1 and R 2 , independently of the other, is hydrogen or substituted or unsubstituted C^_ g alkyl, C 3 _ g alkenyl, C 3 _ g cycloalkyl, C 4 _ 7 cycloalkylalkyl, C 4 _ g cycloalkenyl, C 4 _ 7 cycloalkenylalkyl, C 7 _ 1Q aralkyl, C g _ 10 aralkenyl, or monocyclic heteroar(C^-j alkyl) having 5-6 ring atoms, one or more of which is oxygen, sulphur, and/or nitrogen; in which the ring and/or chain substituent or substituents is or are chlorine, fluorine, hydroxy, C^_ 3 alkoxy, di(C^_ 3 alkyl)amino, or C^_ g alkylthio; and R^ is hydrogen; C 1-g alkyl, C 2 _ g alkenyl; C 2 _ g alkoxyalkyl; C 2 _| 2 alkylaminoalkyl; Cj_ 12 dialkylaminoalkyl; C 1-g perfluoroalkyl; C 2 _ g alkylthioalkyl; or a substituted or unsubstituted phenyl; benzyl; phenethyl; monocyclic heteroaryl, heteroaralkyl, heterocyclyl or heterocyclylalkyl radical in which the substituent or substituents is or are chlorine, fluorine, hydroxy, C 1-3 alkoxy, di(C^_ 3 alkyl) amino or alkylthio. 6. A compound according to Claim 3, in which 1 2 each of R and R , independently of the others, are as defined in Claim 5. 7. A compound according to Claim 5, in which 1 2 each of R and R , independently of the others, is hydrogen, C^_ g alkyl, C 3 _ g alkenyl or benzyl. 8. A compound according to Claim 6, in which each of R 1 and R 2 , independently of the others, is hydrogen, C^_ g alkyl, C 3 _ g alkenyl or benzyl. 9. A compound according to Claim 7, in which R® is hydrogen, methyl, ethyl, isopropyl, methoxymethyl, hydroxyethyl, hydroxymethyl, dimethylaminomethyl, methylaminomethyl, trifluoromethyl, methylthiomethyl, ethyl95 4687 3 thiomethyl, phenyl, benzyl, phenethyl, 2-pyridyl, 3pyridyl, 4-pyridyl, 2-thiazolyl or 4-thiazolyl; and 1 2 each of R and R , independently of the other, is hydrogen, methyl, ethyl, propyl, isopropyl, butyl, t5 butyl, allyl, or benzyl. 10. A compound according to Claim 7, in which T 2 6 each of R , R and R , independently of the others, is hydrogen, C^_g alkyl or C 3 _g alkenyl. 11.i A dompound according to Claim 6, in which 1 2 10 each of R and R , independently of the other, is hydro gen, Cj_g alkyl or θ 3 _ 6 alkenyl. 12. A compound according to Claim 7 having the structure: OH 15 13. A compound according to Claim 10 having the structure: / 14. A compound according to Claim 11 having the structure: sch 2 ch 2 n— C COOH CH n NH NH„ 15. A compound according to Claim 10 having the structure: CH, sch 2 ch 2 nC0 2 CH 2 0CC(CH 3 ) 3 OH wherein A is a pharmaceutically acceptable anionic salt residue. 16. A compound according to Claim 10 having the 10 structure: r J 46873 17. A compound according to Claim 7 having the structure: X NH wherein R is 3-pyridyl. 5 18. A compound according to Claim 7 having the structure: wherein R is 4-thiazolyl. 19. A compound according to Claim 10 having the 10 structure: 20. A compound according to Claim 10 having the structure: Oil f I! 3 sch 2 ch 2 n=c^ \ COO θ H 21. A compound according to Claim 9 having the structure: 22. A compound according to Claim 10 having the structure: sch 2 ch 2 nCOOH CH. N-CH 2 CH=CH 2 10 23. A compound according to Claim 10 having the structure: 24. A compound according to Claim 10 having the structure: OH CH, NCH 2 CH 3 SCH,CH,N— C COOH 5 25. A compound according to Claim 10 having the structure: 26. A compound according to Claim 10 having the structure: 100 27. A compound according to any one of Claims 3

5. 53. A compound as claimed in Claim 1, when prepared by a method substantially as hereinbefore described in any one of Examples 1 to 37.