TITLE OF THE INVENTION

CARBAPENEM COMPOUNDS. COMPOSITIONS AND METHODS OF TREATMENT

BACKGROUND OF THE INVENTION

The present invention relates to antibacterial agents of the carbapenem class in which the five membered pyrrolidine ring of the carbapenem nucleus is substituted by various cationic and neutral -S- heteroaryl substituents.

Thienamycin was an early carbapenem antibacterial agent having a broad spectrum; it has the following formula:

Later, N-formimidoyl thienamycin was discovered; it has the formula:

The carbapenems of the present invention are useful against gram positive microorganisms, especially methicillin resistant Staphylococcus aureus (MRS A), methicillin resistant Staphylococcus epidermidis (MRSE), and methicillin resistant coagulase negative Staphylococci (MRCNS). The antibacterial compounds of the present invention thus comprise an important contribution to therapy of these difficult to control pathogens. There is an increasing need for agents effective against such pathogens (MRSA/MRCNS) which are at the

same time safe, i.e., relatively free from undesirable side effects.

Also, certain carbapenems of the present invention have a relatively low level of inactivation by dehydropeptidase and penicillinase, two enzymes known to reduce the serum levels of conventional beta lactam antibiotics.

SUMMARY OF THE INVENTION

The present invention addresses a compound represented by the structural formula:

wherein:

R ¹ represents hydrogen or methyl; CO ₂ M represents a carboxylic acid, a carboxylate anion with or without a pharmaceutically acceptable counterion, a pharmaceutically acceptable ester group or a carboxylic acid protected by a protecting group;

P* represents hydrogen or a hydroxyl protecting group; Het represents a heterocyclic group which is uncharged or positively charged, with no more than three positive charged atoms, and is selected from the group consisting of:

aanndd

wherein:

- represents the point of attachment to S;

A represents O or S;

X, Y and Z independently represent CR, N or N ⁺ R ^a , provided that for any given compound at least one of X, Y and Z represents CR and no more than one of X, Y and Z represents N ⁺ R ^a : R represents a member selected from the group consisting of hydrogen; halo; -CN; -N0 ₂ ; -NR ^a R ^b ; -OR ^c ; -SR ^C ; -CONR aRb; -COORh; -SOR ^c ; -S( ₂ R ^C ; -Sθ ₂ NR ^a R ^b ; -NR ^a SQzR ^b ; -COR ^a ; -NR ^a COR*> -OCOR ^a ; -OCONR ^a R ^b ; -NR ^a CONR!>R ^c _; -NRaC0 ₂ R"; -OCQ ₂ R"; -C(NR«)NRbRc _; -NRaC(NH)NRbRc _; -NR ^a C(NR ^b )R ^c ; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R ^d groups; -C3-7 cycloalkyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R ^d groups; -C5-7 cycloalkenyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched-chain alkynyl, unsubstituted or substituted with one to four R ^d groups; -Q; -(CH ₂ ) _n Q where n = 1 -4; and -R*;

R ^a , R ^b and R ^c independently represent hydrogen, -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R ^d groups; -C3-7 cycloalkyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched-chain alkynyl, unsubstituted or substituted with one to four R ^d groups, -R* or -(CH2) _n Q where n = 1-3; or R ^a and R ^b taken together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by one or more of O, S, NR ^C , with R ^c as defined above, or -C(O)-, said ring being unsubstituted or substituted with one to four R' groups; or R ^b and R taken together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by one or more of O, S, NR ^a , with R ^a as defined above, or -C(O)-, said ring being unsubstituted or substituted with one to four R ¹ groups;

R represents halo; -CN; -N0 ₂ ; -NR^R ^f ; -OR*=; -SRϋ; -CONR ^e R ^f ; -COORg; -SORε; -S0 ₂ Rs; -S0 ₂ NR*Rf; -NR^SCW; -COR«;

-NR ^« COR ⁱ ; -OCOR ^e ; -OCONR ^e R-; -NR ^e CONKR*^; -NR 0 ₂ R ^h ; -OC0 ₂ R"; -C(NR< ^* )NR ^t R ^j -; -NR ^e C NH)N fR ; -NReC( NRf) ^ -R* or -Q;

R ^e , R ^f and R*. independently represent hydrogen; -Ci- straight- or branched-chain alkyl, either unsubstituted or substituted with one to four R> groups or -R*; C2-6 straight- or branched-chain alkyl, either unsubstituted or substituted with one to four R> groups or -R*; or

R ^e and R ^f taken together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by one or more of O, S, -C(O)- or NRs with R as defined above, said ring being unsubstituted or substituted with one to four R' groups;

R ^h represents hydrogen or a -Ci-6 straight or branched-chain alkyl group or phenyl;

R' represents halo; -OR ^h ; -CN; -NO2; phenyl, 2- imidazolyl, -NHSO2R ¹¹ ; -NH(Rh); -N(Rh) ₂ ; -N+(Rh) ; -C(0)NHR ^h ; -C(0)N(R ^h )2; -Sθ2N(R ^h )2; pyridyl; pyridinium; methyl-imidazolium; -Cθ2R ^h ; -C(0)R ^h ; guanidinyl; carbamimidoyl or ureido;

R* is a member selected from the group consisting of:

d represents -C(O)-, NR ^k , O, or S; E, G, e, g, x, y and z independently represent CR , N or N ⁺ R ^k , provided that no more than one of E, G, e, g, x, y and z represents N ⁺ R ^k ;

R ^k represents hydrogen; -Ci-6 straight- or branched- chain alkyl, unsubstituted or substituted with one to four R' groups and/or Q; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R ^» groups and/or Q; -C2-6 straight- or branched-chain alkynyl, unsubstituted or substituted with one to four R' groups and/or 0, or -(CH ₂ ) _n Q where n = 1-4;

R ^m represents a member selected from the group consisting of: hydrogen; halo; -CN; -N0 ₂ ; -NHR"; -NR ⁿ R°; -OR"; -SR"; -CONR ⁿ R ^o ; -COOR"; -SOR"; -S0 ₂ R"; -S0 ₂ NR"R°; -NR"SQ2 °; -COR ⁿ ; -NR"COR°; -OCOR"; -OCONR ⁿ R"; -NR ⁿ C0 ₂ R ^h ; -NR ^b C0 ₂ R"; -NR"CONRoRh; -OC0 ₂ R ^h ; -CNR"NRoRh _; -NRnC(NH)NR°R ^h ; -NR ⁿ C(NR°)R ^h ; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R- groups and/or Q; -C3-7 cycloalkyl, unsubstituted or substituted with one to four R' groups and/or Q; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R' groups and/or Q; -C5.7 cycloalkenyl, unsubstituted or substi¬ tuted with one to four R' groups and/or Q; -C2-6 straight- or branched- chain alkynyl, unsubstituted or substituted with one to four R- groups and/or Q; phenyl, unsubstituted or substituted with one to four R> groups and/or -(CH ₂ ) _n Q where n = 1 -4; -Q and -(CH ₂ ) _n Q where n = 1-4;

R ⁿ and R° independently represent hydrogen; phenyl, unsubstituted or substituted with one to four R' groups and/or -(CH ₂ ) _n Q where n = 1 -4; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups and/or Q; -C3-7 cycloalkyl, unsubstituted or substituted with one to four R' groups and/or Q; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R' groups and/or 0; -C5- ₇ cycloalkenyl, unsubstituted or substi-

tuted with one to four R ¹ groups and/or Q; -C2-6 straight- or branched- chain alkynyl, unsubstituted or substituted with one to four R> groups and/or Q; or -(CH2)nO where n = 1-4; or R ⁿ and R° taken together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by one or more of O, S, NR ^h , with R ^h as defined above, or -C(O)-, said ring being unsubstituted or substituted with one to four R' groups and/or Q;

0 represents a member selected from the group consisting of:

wherein:

^~ Y represents the point of attachment; a and b independently represent 1 , 2 or 3; α represents NR ^S , O or S; β, δ, λ, μ and σ independently represent CR ¹ , N or N ⁺ R ^s provided that no more than two of β, δ, λ, μ, and σ may be N ⁺ R ^s and that Q as a whole has at least one but not more than three positive charges;

R ^s represents hydrogen; phenyl; -NH ₂ ; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R> groups; -C2- ₆ straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R' groups; or -C2-6 straight- or branched- chain alkynyl, unsubstituted or substituted with one to four R' groups; R ^l represents hydrogen; halo; phenyl; -CN; -N0 ; -NHR"; -NR"R ^V : -OR": -SR": -CONR"R ^v ; -COOR ^h ; -SOR"; -S0 ₂ R";

-S0 ₂ NR"Rv; -NR"S0 ₂ R ; -COR"; -NR"COR ^v ; -OCOR"; -OCONR"R ^v ; -NR"C0 ₂ R ^v ; -NR ^u CONR ^v R ^w ; -OC0 ₂ R ^v ; pyridyl; pyridinium; methyl- pyridinium; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R- groups; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R' groups; -C2-6 straight- or branched-chain alkynyl, unsubstituted or substituted with one to four R' groups;

R ^u and R ^v independently represent hydrogen; -Ci- straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups; or R ^u and R ^v together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by one or more of O, S, NR ^W or -C(O)-, said ring being unsubstituted or substituted with one to four R- groups; R represents hydrogen or -C 1 -6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R- groups; and

R ,R ^y ,and R ^z independently represent hydrogen; phenyl; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups and optionally interrupted by O, S,

NRw, N ⁺ R ^h R ^w or -C(O)-; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R- groups; or -C2-6 straight- or branched-chain alkynyl, unsubstituted or substituted with one to four

R- groups; or R ^x and R together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by O, S, R ^w , N+RhRw or -C(O)-, and, when R ^x and Ry together represent a 4-6 membered ring as described above, R ^z is as described above or R ^z represents an additional saturated 4-6 membered ring fused to the ring represented by

R ^x and R taken together, optionally interrupted by O, S, NR or -C(O)-, said rings being unsubstituted or substituted with one to four R' groups; and

L" represents a pharmaceutically acceptable counterion .

Pharmaceutical compositions, intermediates, processes of manufacture and methods of treatment are also disclosed.

DETAILED DESCRIPTION OF THE INVENTION The invention is described herein in detail using the terms defined below unless otherwise specified. Carboxylic acid refers to -COOH. Carboxylate anion refers to a negatively charged group

-coo-. Carbamoyl and the carbamoyl portion of carbamoyloxy, when referring to a value of R, are defined as -C(0)NR ^a R ^b ; when referring to a value of R ^d , carbamoyl represents -C(0)NR ^e R ^f ; when referring to a value of Ri, carbamoyl is -C(0)NHR ^h or -C(0)N(Rh)2; when referring to a value of R ^m , carbamoyl is -C(0)NR ⁿ R°; and when referring to a value of R-, carbamoyl is -C(0)NR ^u R ^v .

Sulfinyl, when referring to a value of R, represents -S(0)R ^c ; when referring to a value of R ^d , sulfinyl is -S(0)R8; when referring to a value of R ^m , sulfinyl is -S(0)R ⁿ ; and when referring to a value of R-, sulfinyl is -S(0)R ^u . Sulfonyl, when referring to a value of R, represents

-Sθ2R ; when referring to a value of R ^d , sulfonyl is -SO2RS; when referring to a value of R ^m , sulfonyl is -Sθ2R ⁿ ; and when referring to a value of R-, sulfonyl is -SO2R ¹¹ .

Sulfamoyl, when referring to a value of R, represents -Sθ2NR ^a R ^D ; when referring to a value of R ^d , sulfamoyl is -Sθ2NR ^e R ^f ; when referring to a value of R sulfamoyl is -Sθ2N(R ^h )2; when referring to a value of R ^m , sulfamoyl is -Sθ2NR ⁿ R°; and when referring to a value of R ^l , sulfamoyl is -Sθ2NR ^u R . Sulfonamido, when referring to a value of R, represents

-NR ^a Sθ2R ^D ; when referring to a value of R ^d , sufonamido is -NR ^e S02Rf, when referring to a value of R*, sulfonamido is -NHS02R ^h ; when referring to a value of R ^m , sulfonamido is

-NR ⁿ Sθ2R°; and when referring to a value of R ^l , sulfonamido is -NR ^u Sθ2R ^v .

Acyl and the acyl portion of acylamino and acyloxy. when referring to a value of R, represents -C(0)R ; when referring to

5 a value of R ^d , acyl is -C(0)R ^e ; when referring to a value of R ^m , acyl is -C(0)R ⁿ ; and when referring to a value of R-, acyl is C(0)R ^u .

Amidino, when used in connection with the values of R ^d , refers to -C(NR ^e )NR ^f Rg; when used in connection with the values of R ^m , amidino refers to -C(NRn)NRoRh. l o Guanidino, when used in connection with the values of

R ^d , refers to -NR ^e C(NH)NR ^f RS; when used in connection with the values of Rm, guanidino refers to -NRnC(NH)NRoRh.

Carbamimidoyl, when used in connection with the values of R ^d , refers to -NR ^e C(NR ^f )RS; when used in connection with the

15 values of R ^m , carbamimidoyl refers to -NR ⁿ C(NR°)R ^h .

Ureido, when referring to a value of R, represents -NR ^a C(0)NR ^b R ^c ; when referring to a value of R ^d , ureido is -NR ^e C(0)NRfRg; when referring to a value of R ^m , ureido is -NR ⁿ C(0)NR°R ^h ; and when referring to a value of R-, ureido is

20 -NRUC(0)NRVRW.

When the group N ⁺ R ^a is present, such as in the definition of E, G, X, Y and Z, N ⁺ R ^a is a substituted (quatemized) nitrogen atom which has a formal positive charge due to the presence of substituent R .

25 The term "alkyl" refers to a monovalent alkane (hydro¬ carbon) derived radical containing from 1 to 10 carbon atoms unless otherwise defined. It may be straight, branched or cyclic. Preferred alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, t-butyl, cyclopentyl and cyclohexyl. When substituted, alkyl groups may be

30 substituted with up to four substituent groups, R ¹ , at any available point of attachment. When the alkyl group is said to be substituted with an alkyl group, this is used interchangeably with "branched alkyl group".

Cycloalkyl is a specie of alkyl containing from 3 to 15 carbon atoms, without alternating or resonating double bonds between carbon atoms. It may contain from 1 to 4 rings which are fused. The term "alkenyl" refers to a hydrocarbon radical straight, branched or cyclic containing from 2 to 10 carbon atoms and at least one carbon to carbon double bond. Preferred alkenyl groups include ethenyl, propenyl, butenyl and cyclohexenyl.

The term "alkynyl" refers to a hydrocarbon radical straight or branched, containing from 2 to 10 carbon atoms and at least one carbon to carbon triple bond. Preferred alkynyl groups include ethynyl, propynyl and butynyl.

Aryl refers to aromatic rings e.g., phenyl, substituted phenyl and the like, groups as well as rings which are fused, e.g., naphthyl, phenanthrenyl and the like. An aryl group thus contains at least one ring having at least 6 atoms, with up to five such rings being present, containing up to 22 atoms therein, with alternating (resonating) double bonds between adjacent carbon atoms or suitable heteroatoms. The preferred aryl groups are phenyl, naphthyl and phenanthrenyl. Aryl groups may likewise be substituted as defined. Preferred substituted aryls include phenyl and naphthyl.

The term "heteroaryl" refers to a monocyclic aromatic hydrocarbon group having 5 or 6 ring atoms, or a bicyclic aromatic group having 8 to 10 atoms, containing at least one heteroatom, O, S or N, in which a carbon or nitrogen atom is the point of attachment, and in which one or two additional carbon atoms are optionally replaced by a heteroatom selected from O or S, and in which from 1 to 3 additional carbon atoms are optionally replaced by nitrogen heteroatoms. said heteroaryl group being optionally substituted as described herein. Heteroaryl thus includes aromatic and partially aromatic groups which contain one or more heteroatoms. Examples of this type are pyrrole, pyridine, oxazole. thiazole and oxazine. Additional nitrogen atoms may be present together with the first nitrogen and oxygen or sulfur, giving, e.g., thiadiazole.

The term "heterocycloalkyl" refers to a cycloalkyl group (nonaromatic) in which one of the carbon atoms in the ring is replaced by a heteroatom selected from O, S or N, and in which up to three additional carbon atoms may be replaced by said hetero atoms. The term "quaternary nitrogen" refers to a tetravalent positively charged nitrogen atom including, e.g., the positively charged nitrogen in a tetraalkylammonium group (eg. tetramethyl- ammonium, N-methylpyridinium), the positively charged nitrogen in protonated ammonium species (eg. trimethylhydroammonium, N- hydropyridinium), the positively charged nitrogen in amine N-oxides (eg. N-methylmorpholine-N-oxide, pyridine-N-oxide), and the positively charged nitrogen in an N-amino-ammonium group (eg. N-aminopyridinium).

The term "heteroatom" means O, S or N, selected on an independent basis.

Halogen and "halo" refer to bromine, chlorine, fluorine and iodine.

Alkoxy refers to C \ -C4 alkyl-O-, with the alkyl group optionally substituted as described herein. An "alkoxy carbonyl" radical is represented by the formula: -C(0)OR ^h , where the R ^h group is a straight or branched Ci- alkyl group. When referring to R ^d , the group -COORS represents an alkoxycarbonyl group where RS is a C2-6 straight or branched alkyl group. When referring to R ^m and R-, the group -COOR ^h is an alkoxycarbonyl group.

When a group is termed "substituted", unless otherwise indicated, this means that the group contains from 1 to 4 substituents thereon. With respect to R, R ^a , R ^b and R ^c , the substituents available on the alkyl, alkenyl and alkynyl groups are selected from the values of R ^d . Many of the variable groups are optionally substituted with up to four R- groups. With respect to R ^e , R ^f and RS, when these variables represent substituted alkyl, the substituents available thereon are selected from the values of R>. With respect to R ^k , when R is a substituted alkyl, alkenyl or alkynyl group, the substituents available

thereon are selected from the values of R ¹ . With respect to R ^m , when R ^m represents a substituted straight or branched alkyl, cycloalkyl, alkenyl, cycloalkenyl or alkynyl group, the subtituents available thereon are selected from the values of RK When R ⁿ , R°, R ^u , R ^v or

5 R represents a substituted alkyl group, the substituents present thereon are selected from the values of R*. Lastly, with respect to R ^s , R-, R , Ry and R ^z , when these variables represent a substituted alkyl, alkenyl or alkynyl group, the substituents present thereon are selected from the values of R*. l o When a functional group is termed "protected", this means that the group is in modified form to preclude undesired side reactions at the protected site. Suitable protecting groups for the compounds of the present invention will be recognized from the present application taking into account the level of skill in the art, and

15 with reference to standard textbooks, such as Greene, T. W. et al. Protective Groups in Organic Synthesis Wiley, New York (1991 ). Examples of suitable protecting groups are contained throughout the specification.

A subset of compounds of the present invention includes

20 compounds with a positively charged (cationic) atom, balanced by a negatively charged carboxylate anion, and when necessary for charge balance, such as in bis-cationic compounds, an additional negatively charged counterion or counterions. It is noted that the overall molecule contains no more than three positively charged

25 atoms. Thus, when it is said that Het contains no more than three positively charged atoms, this includes positively charged substituent groups attached to Het.

Of the charged compound subset, one group of compounds is comprised of those which contain a positively charged

30 atom in the Het ring. In these compounds, R - and P* are as previous¬ ly defined; CO2M is a carboxylate anion (M represents a negative charge). Within the definition of -Het, one of X, Y and Z represents N ⁺ R ^a , with R ^a as previously defined.

Another group of compounds within this subset is comprised of compounds which contain a positively charged moiety on a group which is attached to the -Het group. For example, when X,

Y or Z represents CR, and R represents -R*, one of the values of e, g, 5 x, y and z is N+R ^k with R ^k as previously defined.

Yet another group of compounds within this subset is comprised of compounds wherein one of e, g, x, y and z represents CR ^m , and R ^m represents 0 or -(CH2)n-Q. In these compounds, one of the values of β, δ, λ, μ and σ is N ⁺ R ^s , and R ^s is as previously defined, l o Another subset of compounds of formula I is comprised of compounds in which -Het and groups attached thereto are uncharged. In these compounds, P*, CO2M and R ¹ are as previously defined with respect to formula I. Within Het, A represents O or S; X,

Y and Z represent CR or N. R is as previously defined except that it 15 does not represent a positively charged moiety. Thus, none of e, g, x, y, z, β, δ, λ, μ and σ represents a quaternary nitrogen atom N+R ^k or N+Rs.

Preferred compounds of the invention include those compounds of formula I wherein: 20 R ¹ represents hydrogen or methyl;

C0 ₂ M represents a carboxylic acid or a carboxylate anion with or without a pharmaceutically acceptable counterion;

P* represents hydrogen;

Het represents a heterocyclic group which is positively 25 charged, with no more than three positive charged atoms, and is selected from the group consisting of:

^■ k ' II

-Y

30 wherein:

^■ 5- represents the point of attachment to S; A represents S; Z represents N:

X and Y represent CR or N, provided that for any given compound at least one of X and Y represents CR;

R represents a member selected from the group consisting of hydrogen; halo; -CN; -N0 ₂ ; -CONR R ^b ; -COOR ^h ; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R ^d groups; -C3-7 cycloalkyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R ^d groups; -C5-7 cycloalkenyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched- chain alkynyl, unsubstituted or substituted with one to four R ^d groups; -(CH ₂ ) _n Q where n = 1 -4; and -R*;

R ^a and R ^b independently represent hydrogen, -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R ^d groups; -C3-7 cycloalkyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R ^d groups; -C2-6 straight- or branched- chain alkynyl, unsubstituted or substituted with one to four R ^d groups, -R* or -(CH ₂ ) _n Q where n = 1-3; or R ^a and R ^b taken together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by one or more of O, S, NR ^C , with R ^c as defined above, or -C(O)-, said ring being unsubstituted or substituted with one to four R' groups;

R ^d represents halo; -CN; -NReR ^f ; -OR ^g ; -CONR?R ^f ; -COOR ^g ; -COR ^e ; -R* or -Q; R ^e , R ^f and Rs independently represent hydrogen; -C 1 -6 straight- or branched-chain alkyl, either unsubstituted or substituted with one to four R' groups or -R*; C2-6 straight- or branched-chain alkyl, either unsubstituted or substituted with one to four R' groups or -R*; or R ^e and R ^f taken together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by one or more of O, S, -C(O)- or NRs with RS as defined above, said ring being unsubstituted or substituted with one to four R' groups;

R ^h represents hydrogen or a straight or branched-chain alkyl group or phenyl;

R- represents halo; -OR ^h ; -CN; -NO2; phenyl, 2- imidazolyl; -NHSO2R ¹¹ ; NH(R ^h ); N(Rh) ₂ ; N ⁺ (R ^h )3; C(0)NHR ^h ; C(0)N(R ^h )2; Sθ2N(R ^h )2; pyridyl; pyridinium; methyl-imidazolium; Cθ2R ^h ; C(0)R ^h ; guanidinyl; carbamimidoyl or ureido;

R* is a member selected from the group consisting of:

wherein:

^~ ^ ^~ represents the point of attachment;

d represents -C(O)-, O, or S; e, g, x, y and z independently represent CR ^m , N or N ⁺ R ^k , provided that no more than one of e, g, x, y and z represents N ⁺ R ^k ;

R ^k represents hydrogen; -Ci-6 straight- or branched- chain alkyl, unsubstituted or substituted with one to four R- groups and/or 0; or -(CH ₂ ) _n Q where n = 1-4;

Rm represents a member selected from the group consisting of: hydrogen; halo; -CN; -N0 ₂ ; -NHR"; -NR"R°; -OR"; -SR"; -CONR ⁿ R ^o ; -COOR"; -NR ⁿ S0 ₂ R ^» ; -COR"; -NR"COR°; -NR"C0 ₂ R ^h ; -NR ^b C0 ₂ R"; -NR"CONRoRh _; -CNR"NR°Rh; -NR"C(NH)NR°R ^b ; -NR ⁿ C(NR°)R ^h ; -Cj-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R- groups and/or Q; -C3.7 cycloalkyl, unsubstituted or substituted with one to four R' groups and/or Q; -C2-6 straight- or branched-chain alkenyl. unsubstituted or substituted with one to four R' groups and/or Q; -C5.7 cycloalkenyl, unsubstituted or substituted with one to four R' groups and/or Q; phenyl, unsubstituted or substituted with one to four R' groups and/or -(CH ₂ ) _n Q where n = 1 -4; -0 and -(CH ₂ )„Q where n = 1 -4;

R" and R° independently represent hydrogen; phenyl, unsubstituted or substituted with one to four R' groups and/or -(CH ₂ ) _n Q where n = 1-4; -Ci-6 straight- or branched-chain alkyl, unsubstituted or

substituted with one to four R ¹ groups and/or Q; -C3.7 cycloalkyl, unsubstituted or substituted with one to four R- groups and/or Q; or -(CH2)nO where n = 1-4; or R ⁿ and R° taken together with any intervening atoms represent a 5-6 membered saturated ring optionally interrupted by one or more of O, S, NR ^h , with R ^h as defined above, or -C(O)-, said ring being unsubstituted or substituted with one to three R ^» groups and/or Q;

0 represents a member selected from the group consisting of:

wherein:

-5- represents the point of attachment; a and b independently represent 2 or 3; α represents NR ^S , O or S; β, δ, λ, μ and σ independently represent CR ¹ , N or N ⁺ Rs provided that no more than two of β, δ, λ, μ, and σ may be N ⁺ R ^s and that Q as a whole has at least one but not more than three positive charges;

R ^s represents hydrogen; phenyl; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups; or -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R' groups;

R ^l represents hydrogen; halo; -CN; -N0 ₂ ; -NHR ^U ; -NR ^U R ^V ; -OR"; phenyl, pyridyl; pyridinium; methyl-pyridinium; -C _\ . , straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups;

R" and R ^v independently represent hydrogen: -Ci-f, straight- or branched-chain alkyl, unsubstituted or substituted with one to four R ⁱ groups;

Rw represents hydrogen or -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups;

R ^x ,Ry ,and R ^z independently represent hydrogen; phenyl; -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups and optionally interrupted by O, S, NR , N+RhRw or -C(O)-; -C2-6 straight- or branched-chain alkenyl, unsubstituted or substituted with one to four R- groups; or -C2-6 straight- or branched-chain alkynyl, unsubstituted or substituted with one to four R' groups; or R and R ^y together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by O, S, NR , N+RhRw or -C(O)-, and, when R ^x and Ry together represent a 4-6 membered ring as described above, R ^z is as described above or R ^z represents an additional saturated 4-6 membered ring fused to the ring represented by R and Ry taken together, optionally interrupted by O, S, NR ^W or -C(O)-, said rings being unsubstituted or substituted with one to four R' groups; and

L" represents a pharmaceutically acceptable counterion . More preferred compounds of the invention are those falling within formula I wherein: R ¹ represents methyl;

C0 ₂ M represents a carboxylic acid or a carboxylate anion with or without a pharmaceutically acceptable counterion; P* represents hydrogen;

Het represents a heterocyclic group which is positively charged, with no more than two positive charged atoms, and is selected from the group consisting of:

wherein:

^~ ^ ^~~ represents the point of attachment to S;

X and Y independently represent CR

R represents a member selected from the group consisting of hydrogen; halo; -CN; -N0 ₂ ; C1-3 straight- or branched- chain alkyl, unsubstituted or substituted with one R ^d group; and -R*;

R ^d represents -R*

R ^h represents hydrogen or a -Ci-6 straight or branched-chain alkyl group or phenyl;

R- represents halo: -OR ^h ; -CN; -NO2; phenyl, 2- imidazolyl; -NHS02R ^h ; NH(R ^h ); N(R ^h )2; N ⁺ (R ^h )3; C(0)NHRh; C(0)N(R ^h )2; Sθ2N(R ^h )2; pyridyl; pyridinium; methyl-imidazolium; CO2R ¹¹ ; C(0)R ^h ; guanidinyl; carbamimidoyl or ureido;

R* is a member selected from the group consisting of:

wherein

-5- represents the point of attachment;

d represents S; e, g, x, y and z independently represent CR ^m or N;

R ^m represents a member selected from the group consisting of: hydrogen; halo; -CN; -C ₁ -4 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R' groups and/or Q; and -(CH ₂ ) _n Q where n = 1 -4; 0 represents a member selected from the group consisting of:

wherein:

X-

^J represents the point of attachment;

R ^s represents hydrogen; or -C1-3 straight-chain alkyl, unsubstituted or substituted with one or two R* groups;

R represents hydrogen or -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R ¹ groups;

R ^x ,R ^y ,and R ^z independently represent hydrogen; phenyl; or -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to three R' groups and optionally interrupted by O, S, NRw, N+RhRw _or -C(O)-; or R and R ^y together with any intervening atoms represent a 4-6 membered saturated ring optionally interrupted by O, S, NR ^W , N+RhRw or -C(O)-, and, when R ^x and Ry together represent a 4-6 membered ring as described above, R ^z is as described above or R ^z represents an additional saturated 4-6 membered ring fused to the ring represented by R ^x and Ry taken together, optionally interrupted by O, S, NR or -C(O)-, said rings being unsubstituted or substituted with one to four R- groups; and

L" represents a pharmaceutically acceptable counterion .

The most preferred compounds of the invention include those compounds falling within formula I wherein:

R ¹ represents methyl;

C0 ₂ M represents a carboxylate anion with or without a pharmaceutically acceptable counterion;

P* represents hydrogen;

Het represents a heterocyclic group which is positively

charged, with one or two positive charged atoms, and is selected from the group consisting of:

wherein:

-* represents the point of attachment to S;

X and Y independently represent CR

R represents a member selected from the group consisting of hydrogen; halo; -CN; and -R*;

R ^h represents hydrogen or a -C1-3 straight-chain alkyl group;

R' represents halo; -OR ^h ; -CN; -NO2; phenyl, 2- imidazolyl; -NHS0 ₂ R ^h ; NH(R ^h ); N(Rh) ₂ : N+(Rh) ₃ ; C(0)NHRh _; C(0)N(R ^h )2; Sθ2N(R ^h )2; pyridyl; pyridinium; methyl-imidazolium; CO2R ¹¹ ; C(0)R ^h ; guanidinyl; carbamimidoyl or ureido;

R* is a member selected from the group consisting of:

wherein

represents the point of attachment;

e, g, x, y and z independently represent CR ^m ; R represents a member selected from the group consisting of: hydrogen; halo; -CN; and -(CH ₂ ) _n Q where n = 1-4;

0 represents a member selected from the group consisting of:

wherein:

^■ 5- represents the point of attachment;

R ^s represents hydrogen or C1-3 straight-chain alkyl, unsubstituted or substituted with one to two R- groups;

R ^w represents hydrogen or -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to four R ¹ groups;

R ^x ,R ^y ,and R ^z independently represent hydrogen; or -Ci-6 straight- or branched-chain alkyl, unsubstituted or substituted with one to three R' groups and optionally interrupted by O, S, NR , N ⁺ R ^h R ^w or -C(O)-; or R and R ^y together with any intervening atoms represent a 5-6 membered saturated ring optionally interrupted by O, S, NR ^W , N ⁺ RhRw or -C(O)-, and, when R ^x and Ry together represent a 4-6 membered ring as described above, R ^z is as described above or R ^z represents an additional saturated 4-6 membered ring fused to the ring represented by R ^x and R ^y taken together, optionally interrupted by O, S, NR or -C(O)-, said rings being unsubstituted or substituted with one to four R' groups; and

L" represents a pharmaceutically acceptable counterion . The compounds of the invention can be synthesized in accordance with the following general schemes and examples. The starting materials for the initial stage of the foregoing sequence are either known in the art or can be readily prepared by established methods.

The compounds of the present invention are prepared by reacting a suitably protected carbapen-2-em -3-carboxylate having

a suitable leaving group at the 2-position with a heterocyclic thiol (mercaptan) under basic conditions, modifying the thus-introduced side chain (if desired), and then removing any protecting groups which are present to afford the desired final product. The process is illustrated by the following generic scheme:

REACTION SCHEME A

p

With reference to Reaction Scheme A, P*, R- , Het, M, and Q are as defined with respect to the compounds of formula I.

P represents a carboxyl protecting group.

Het* represents a heterocyclic group selected from the group defined for Het above, but is modified in the reaction and no longer identical to the initial Het group.

LG represents a suitable leaving group such as trifluoro- methanesulfonate (triflate), diethyl phosphate, diphenyl phosphate, di-(p-chlorophenyl) phosphate, methanesulfonate (mesylate), benzene- sulfonate, p-toluenesulfonate, chloride, bromide, iodide and the like.

AR represents a suitable alkylating reagent, such as methyl iodide, methyl bromide, benzyl trichloroacetimidate, methyl trifluoro- methanesulfonate, triethyloxonium tetrafluoroborate and the like. In some of the carbapenem compounds of the present invention, M is a readily removable carboxyl protecting group. Such conventional groups consist of known groups which are used to protectively block the carboxyl group during the synthesis procedures described therein. These conventional blocking groups are readily removable, i.e., they can be removed, if desired, by procedures which will not cause cleavage or other disruption of the remaining portions of the molecule. Such procedures include chemical and enzymatic hydrolysis, treatment with chemical reducing or oxidizing agents under mild conditions, treatment with a transition metal catalyst and a nucleophile and catalytic hydrogenation. Examples of such ester forming protecting groups include benzhydryl, p-nitrobenzyl (PNB), 2-naphthylmethyl, allyl, benzyl, trichloroethyl, silyl such as trimethyl- silyl (TMS) or 2-trimethylsilylethyl, phenacyl, p-methoxybenzyl, acetonyl, o-nitrobenzyl, p-methoxyphenyl, 4-pyridylmethyl, and t-butyl. Furthermore, the C-6 hydroxyethyl group of the carbapenem is optionally protected with a hydroxyl protecting group such as trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldimethylsilyl (TBDMS), tert-butyldiphenylsilyl (TBDPS), t-butylmethoxyphenylsilyl, t-butoxydiphenylsilyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxy- carbonyl, benzyloxycarbonyl, t-butyloxycarbonyl, 2,2,2 -trichloroethyl- oxycarbonyl, allyloxycarbonyl, acetyl, 2-trimethylsilylethoxycarbonyl, and the like.

The heterocyclic thiols used in the synthesis of the compounds of the present invention are, in many cases, commercially available compounds. In other cases, the necessary thiols are compounds which have been previously described in the chemical literature; such compounds can be readily prepared by following the procedures described in the literature. In cases where the requisite thiol is neither commercially available nor known in the literature it is necessary to

synthesize the thiol by a newly developed synthesis. Since heterocyclic thiols are well known in the chemical literature, one skilled in the art can, in many cases, adapt a previously published synthesis of an analogous thiol to prepare the requisite thiol in a straightforward manner without undue experimentation. For example, 2-mercapto-benzothiazoles can be readily prepared from commercially available anilines or nitrobenzenes with the appropriate substitution (Comprehensive Heterocyclic Chemistry Volume 6; K.T. Potts, Ed: Pergamon Press, Oxford, 1984). Other mercapto-heterocycles, such as mercapto thiazolopyridines, mercapto- benzoxazoles, mercaptothiazolothiophenes, and the like can be prepared by similar methods well known to those skilled in the art.

One skilled in the art will realize that in compounds where the carbon bearing the mercaptan functionality is adjacent to a nitrogen atom, the compound may exist as an equilibrium mixture of "thiono" and "thiol" tautomers (as shown below) and may, in fact, exist predominantly in the thiono form. However, on treatment with base, the equilibrium will normally shift to favor the salt of the thiol form which can then react with the carbapenem as described below.

"thiono" "thiol"

The addition of the thiol HSHet to the carbapenem is accomplished by treating a solution of the thiol in a suitable solvent such as tetrahydrofuran (THF), ether, acetonitrile, dimethylfoπnamide (DMF), benzene, dimethylsulfoxide (DMSO), dimethoxyethyane, dioxane and the like, with a suitable base such as sodium hydride, sodium hydroxide, lithium hydride, lithium hydroxide, lithium trimethylsiloxane, lithium hexamethyldisilazide, potassium hydride, butyl lithium, methyl lithium, cesium hydroxide, and the like at a temperature between about -20°C and 35°C for about 1 to 90 minutes then combining the carbapenem, either as a solid or in solution, with the resulting mixture.

Alternatively, HSHet. base and carbapenem can be mixed together with a suitable solvent without pre-treatment of the thiol with base, although pre-treatment is preferred. Once the thiol, base, and carbapenem have been mixed, the reaction is allowed to proceed at a temperature between about -20°C and 95 °C for about 0.5 to 24 hours. The crude 2-heteroarylthio substituted carbapenem is puri¬ fied by crystallization or by chromatography on silica gel, and eluted with a suitable solvent or mixture of two or more solvents, such as hexane, ethyl acetate, ether, benzene, dichloromethane, chloroform, acetone, methanol and the like.

For those compounds where further modification of the side chain is not desired, generally those compounds which contain a neutral (uncharged) side chain, the synthesis is completed by removing any protecting groups from the carbapenem hydroxyethyl and carboxy groups, to produce A^ . The term "side chain" as used in this discussion refers to the heterocyclic group which is linked via a sulfur atom to the carbapenem nucleus. The deprotected final product is then purified, if necessary. For ease of handling, the final product, if not crystalline, may be lyophilized from water to afford an amorphous, easily handled solid. The purified product may be characterized structurally by standard techniques, such as nuclear magnetic resonance spectroscopy (NMR), infrared spectroscopy (IR), ultraviolet spectroscopy (UV) and mass spectrum (MS).

For compounds A5 where further modification of the heteroaryl side chain is desired, such as those compounds containing a charged side chain, modifications are best accomplished before removal of the protecting groups. For compounds which contain a hydroxyl group in the side chain, a positive charge may be introduced into the side chain by first activating the hydroxyl group by converting it to a suitable leaving group such as a triflate, mesylate, tosylate, iodide, chloride, bromide, and the like, and then displacing the resulting leaving group with a compound Q, such as N-methyl-imidazole, N-methyl- diazabicyclooctane, N-carbamoylmethyl-diazabicyclooctane, pyridine, N-methylmorpholine and the like which contains a nitrogen atom that

can act as a nucleophile.

In some cases, activation of the hydroxyl group and displacement by Q to produce A4 may be accomplished in a single step by taking advantage of the basic character of compound Q and using it as a base in the activation reaction.

The conversion of the hydroxyl group to a suitable leaving group is accomplished by treating the hydroxyl substituted compound in a suitable solvent with an activating reagent, such as trifluoromethane- sulfonic anhydride, methanesulfonic anhydride, toluene sulfonic anhydride, methanesulfonyl chloride, benzenesulfonyl chloride, toluenesulfonyl chloride, and the like in the presence of a suitable base such as triethylamine, tributylamine, diisopropylethylamine, pyridine, 2,6-lutidine and the like at a temperature between about -78°C and 0°C for about 15 to 120 minutes. The intermediate thus obtained contains a leaving group, which may be made still more reactive to displacement by conversion to the even better leaving group, iodide, by treating a solution of the intermediate in a suitable solvent such as acetone, methyl ethyl ketone and the like, at about -10°C to 50°C with an excess of sodium iodide for about 0.25 to 24 hours. In many cases, the iodide is obtained in sufficiently pure form that it may be used without further purification. For ease of handling, the iodide, if not crystalline, may be lyophilized from benzene to afford an amorphous, easily handled, solid.

The activated hydroxyl group or iodide is displaced by reacting the activated intermediate with reagent Q*. In cases where Q* is sufficiently reactive to displace an activated hydroxyl group (thus render¬ ing further conversion to the iodide unnecessary), activation and displace¬ ment of the hydroxyl group is accomplished in a single step. The activat¬ ing reagent is added to a solution of the hydroxyl substituted compound and a compound Q* (1.0 to 7.5 molar equivalents) in a suitable solvent such as THF, ether, DMF, benzene, acetonitrile, DMSO, and the like at a temperature between about -78 °C and 50 °C for about 15 to 240 minutes.

In cases where the displacement reaction is best accomplished by using the more reactive iodide, a solution of the iodide

is combined with compound Q* ( 1.0 to 7.5 molar equivalents). A silver salt of a non-nucleophilic acid, such as silver trifluoromethanesulfonate, silver tetrafluoroborate and the like is then added. The resulting mixture is then subjected to a standard work-up procedure familiar to those skilled in the art to afford a crude product which is purified, if necessary, by recrystallization or chromatography.

An alternative method for introducing a positive charge into the side chain may be applied to side chains that contain a nitrogen atom which may be quatemized by reaction with a suitable alkylating reagent AR, such as methyl iodide, methyl bromide, benzyl trichloroacetimidate, methyl trifluoromethanesulfonate, triethyloxonium tetrafluoroborate, and the like. Quaternization of the nitrogen atom in the side chain is effected by treating a solution of the compound with a slight excess (1.05 to 1.2 molar equivalents) of the alkylating reagent. The synthesis of the target compound is completed by removing any protecting groups which are present in the penultimate intermediate. The deprotected final product is then purified, as necessary.

The final product may be characterized structurally by standard techniques. For ease of handling, the final product, if not crystalline, may be lyophilized from water to afford an amorphous, easily handled solid.

The carbapenem compounds of the present invention are useful per se and in their pharmaceutically acceptable salt and ester forms in the treatment of bacterial infections in animal and human subjects. The term "pharmaceutically acceptable ester, salt or hydrate," refers to those salts, esters and hydrated forms of the compounds of the present invention which would be apparent to the pharmaceutical chemist, i.e., those which are substantially non-toxic and which may favorably affect the pharmacokinetic properties of said compounds, their palatability, absorption, distribution, metabolism and excretion. Other factors, more practical in nature, which are also important in the selection, are cost of the raw materials, ease of crystallization, yield, stability, hygroscopicity, and flowability of the

resulting bulk drug. Conveniently, pharmaceutical compositions may be prepared from the active ingredients in combination with pharma¬ ceutically acceptable carriers. Thus, the present invention is also concerned with pharmaceutical compositions and methods of treating bacterial infections utilizing as an active ingredient the novel carbapenem compounds.

The pharmaceutically acceptable salts referred to above may also include non-toxic acid addition salts. Thus, the Formula I compounds can be used in the form of salts derived from inorganic or organic acids. Included among such salts are the following: acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, cyclopentane- propionate, digluconate, dodecylsulfate, ethanesulfonate, fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate, hexa- noate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxyethane- sulfonate, lactate, maleate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenyl- propionate, picrate, pivalate, propionate, succinate, tartrate, thiocyanate, tosylate, and undecanoate. With respect to -CO2M, which is attached to the carbapenem nucleus at position 3, this represents a carboxylic acid group (M represents H), a carboxylate anion (M represents a negative charge), a pharmaceutically acceptable ester (M represents an ester forming group) or a carboxylic acid protected by a protecting group (M represents a carboxyl protecting group). The pharmaceutically acceptable salts referred to above may take the form -COOM, where M is a negative charge, which is balanced by a counterion, e.g., an alkali metal cation such as sodium or potassium. Other pharma¬ ceutically acceptable counterions may be calcium, magnesium, zinc, ammonium, or alkylammonium cations such as tetramethyl- ammonium, tetrabutylammonium, choline, triethylhydroammonium, meglumine, triethanolhydroammonium, etc.

The pharmaceutically acceptable esters of the present invention include, for example, those described in detail in U.S. Pat.

No. 4,309,438. Included within such pharmaceutically acceptable esters are those which are hydrolyzed under physiological conditions, such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl, and others described in detail in U.S. Pat. No. 4,479,947. These are also referred to as "biolabile esters".

Biolabile esters are biologically hydrolizable, and many are suitable for oral administration, due to good absorption through the stomach or intestinal mucosa, resistance to gastric acid degradation and other factors. Examples of biolabile esters include compounds in which M represents an alkoxyalkyl, cycloalkoxyalkyl, alkenyloxyalkyl, aryloxyalkyl, alkoxyaryl, alkylthioalkyl, cycloalkyl- thioalkyl, alkenylthioalkyl, arylthioalkyl or alkylthioaryl group. All of these groups can be substituted in the alkyl or aryl portions thereof with acyl or halo groups. The following M species are examples of biolabile ester forming moieties.: acetoxymethyl, 1-acetoxyethyl, 1 - acetoxypropyl, pivaloyloxymethyl, 1-isopropyloxycarbonyloxyethyl, 1-cyclohexyloxycarbonyloxyethyl, phthalidyl and (2-oxo-5 -methyl - 1 ,3-dioxolen-4-yl)methyl.

L- can be present or absent, as necessary to maintain the appropriate charge balance. When present, L" represents a pharma¬ ceutically acceptable counterion. Most anions derived from inorganic or organic acids are suitable. Representative examples of such counterions are the following: acetate, adipate, aminosalicylate, anhydromethylene- citrate, ascorbate, aspartate, benzoate, benzenesulfonate, bromide, citrate, camphorate, camphorsulfonate, chloride, estolate, ethanesulfonate, fumarate, glucoheptanoate, gluconate, glutamate, glycerophosphate, glycolate, 2-hydroxyethanesulfonate, iodide, lactate, lactobionate, malate, maleate, mandelate, methanesulfonate, pantothenate, pectinate, phosphate/diphosphate, polygalacturonate, propionate, salicylate, stearate, succinate, sulfate, tartrate, and tosylate. Other suitable anionic species will be apparent to the ordinarily skilled chemist.

Likewise, when L- represents a specie with more than one negative charge, such as malonate, tartrate or ethylenediaminetetraacetate (EDTA), an appropriate number of carbapenem molecules can be found

in association therewith to maintain the overall charge balance and neutrality.

The compounds of the present invention are valuable antibacterial agents active against various Gram-positive and to a lesser extent Gram-negative bacteria, and accordingly find utility in human and veterinary medicine.

Many of compounds of the present invention are biologically active against MRSA/MRCNS. In vitro antibacterial activity determined in accordance with a standard antibacterial protocol is predictive of in vivo activity, when the compounds are administered to a mammal infected with a susceptible bacterial organism.

Using standard susceptibility tests, the compounds of the invention are determined to be active against MRSA. The compounds of the invention can be formulated in pharmaceutical compositions by combining the compound with a pharmaceutically acceptable carrier. Examples of such carriers are set forth below.

The compounds may be employed in powder or crystalline form, in liquid solution, or in suspension. They may be administered by a variety of means; those of principal interest include: topically, orally and parenterally by injection (intravenously or intramuscularly).

Compositions for injection, a preferred route of delivery, may be prepared in unit dosage form in ampules, or in multidose containers. The injectable compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain various formulating agents. Alternatively, the active ingredient may be in powder (lyophillized or non-lyophillized) form for reconstitution at the time of delivery with a suitable vehicle, such as sterile water. In injectable compositions, the carrier is typically comprised of sterile water, saline or another injectable liquid, e.g., peanut oil for intramuscular injections. Also, various buffering agents, preservatives and the like can be included.

Topical applications may be formulated in carriers such as hydrophobic or hydrophilic bases to form ointments, creams, lotions, in aqueous, oleaginous or alcoholic liquids to form paints or in dry diluents to form powders. Oral compositions may take such forms as tablets, capsules, oral suspensions and oral solutions. The oral composions may utilize carriers such as conventional formulating agents, and may include sustained release properties as well as rapid delivery forms. The dosage to be administered depends to a large extent upon the condition and size of the subject being treated, the route and frequency of administration, the sensitivity of the pathogen to the particular compound selected, the virulence of the infection and other factors. Such matters, however, are left to the routine discretion of the physician according to principles of treatment well known in the anti- bacterial arts. Another factor influencing the precise dosage regimen, apart from the nature of the infection and peculiar identity of the individual being treated, is the molecular weight of the compound. The compositions for human delivery per unit dosage, whether liquid or solid, may contain from about 0.01 % to as high as about 99% of active material, the preferred range being from about 10-60%. The composition will generally contain from about 15 mg to about 2.5 g of the active ingredient; however, in general, it is prefer¬ able to employ dosage amounts in the range of from about 250 mg to 1000 mg. In parenteral administration, the unit dosage will typically include the pure compound in sterile water solution or in the form of a soluble powder intended for solution, which can be adjusted to neutral pH and isotonic.

The invention described herein also includes a method of treating a bacterial infection in a mammal in need of such treatment comprising administering to said mammal a compound of formula I in an amount effective to treat said infection.

The preferred methods of administration of the Formula I antibacterial compounds include oral and parenteral, e.g., i.v. infusion, i.v. bolus and i.m. injection.

For adults, about 5-50 mg of Formula I antibacterial compound per kg of body weight given one to four times daily is preferred. The preferred dosage is 250 mg to 1000 mg of the antibacterial given one to four times per day. More specifically, for mild infections a dose of about 250 mg one to three times daily is recommended. For moderate infections against highly susceptible gram positive organisms a dose of about 500 mg two to four times daily is recommended. For severe, life-threatening infections against organisms at the upper limits of sensitivity to the antibiotic, a dose of about 1000-2000 mg two to four times daily may be recommended. For children, a dose of about 5-25 mg/kg of body weight given 2, 3, or 4 times per day is preferred; a dose of 10 mg/kg is recommended.

The compounds of Formula I are of the broad class known as carbapenems. Many carbapenems are susceptible to attack by a renal enzyme known as dehydropeptidase (DHP). This attack or degradation may reduce the efficacy of the carbapenem antibacterial agent. Many of the compounds of the present invention, on the other hand, are less subject to such attack, and therefore may not require the use of a DHP inhibitor. However, such use is optional and contemplated to be part of the present invention. Inhibitors of DHP and their use with carbapenems are disclosed in, e.g., [European Patent Application Nos. 79102616.4, filed July 24, 1979 (Patent No. 0007 614); and 82107174.3, filed August 9, 1982 (Publication No. 0072 014)].

The compounds of the present invention may, where DHP inhibition is desired or necessary, be combined or used with the appropriate DHP inhibitor as described in the aforesaid patents and published application. The cited European Patent Applications define the procedure for determining DHP susceptibility of the present carbapenems and disclose suitable inhibitors, combination compositions and methods of treatment. A preferred weight ratio of Formula I compound: DHP inhibitor in the combination compositions is about 1 : 1.

A preferred DHP inhibitor is 7-(L-2-amino-2-carboxy- ethylthio)-2-(2,2-dimethylcyclopropanecarboxamide)-2-hepteno ic acid or a useful salt thereof.

The invention is further described in connection with the following non-limiting examples.

PREPARATIVE EXAMPLE 1 4-(4-hvdroxymethylphenvD-2-mercaptothiazole

Step 1

Ethyl 4-chloroacetylbenzoate

A solution of ethyl 4-acetylbenzoate (1.00 g, 5.20 mmol) in glacial acetic acid (5 mL) and concentrated hydrochloric acid (3 mL) was bubbled with chlorine gas at room temperature. After approximately 5 min, a white solid precipitate formed and chlorine addition was stopped.

The reaction mixture was immediately diluted with diethyl ether (50 mL) and washed with water (2 x 50 mL), I M pH7 phosphate buffer, brine, dried over magnesium sulfate, filtered, and evaporated under vacuum to give the title compound (1.09 g) as a white solid.

-H NMR (CDCI3) δ 1.43 (t, CH3), 4.43 (q, OCH2), 4.72 (s, CH2CI),

7.95-8.2 (m, ArH)

MS m/z 227.1 (M + 1 )

Step 2 4-(4-ethoxycarbonylphenylV2-mercaptothiazole

A solution of ethyl 4-chloroacetylbenzoate (500mg, 2.20 mmol) and ammonium dithiocarbamate (267 mg, 2.42 mmol) in water

(4 mL) and tetrahydrofuran (4 mL) was stirred at room temperature. After stirring 16 hrs a white precipitate formed. The solvent was decanted and the solid washed with water (2 x 5 mL), dissolved in ethyl acetate (40 mL) and washed with water (40 mL), brine, dried over magnesium sulfate, filtered, and evaporated under vacuum to give a white solid. The solid was mixed with ethyl acetate (5 mL), filtered, and vacuum dried to afford the title compound (225 mg) as a white solid. lH NMR (CDC13) δ 1.33 (t, CH3), 4 .31 (q, OCH2), 7.52 (s, thiazole H5), 7.87-8.05(m, ArH)

10

Step 3 4-(4-hvdroxymethylphenyD-2-mercaptothiazole

A solution of 4-(4-ethoxycarbonylphenyl)-2-mercapto- _{x 5} thiazole (200 mg, 0.754 mmol) in tetrahydrofuran (3 mL) was treated at room temperature with a solution of lithium aluminum hydride in tetrahydrofuran (IM, 750 μmL, 0.750 mmol) and heated in an oil bath at 60°C. After 30 min the reaction was removed from the bath and allowed to cool to room temperature. The reaction was treated with hydrochloric

₂₀ acid (2N, 0.75 mL), diluted with ethyl acetate (7 mL), and washed with brine (twice), dried over magnesium sulfate, filtered, and evaporated under vacuum to a white solid (168 mg). The crude product was crystallized from hot ethyl acetate (ca. 10 mL) to afford the title compound as an off-white solid (55 mg).

25 -H NMR (DMSOdό) δ 4.51(d, J=5.7 Hz, CH2), 5.28(t, J=5.7 Hz. OH), 7.27 (s, thiazole H5), 7.37 (d, J=8.2 Hz, ArH), 7.69 (d, 2 H, J = 8.2 Hz, ArH) MS m/z 224.0 (M + l )

30

PREPARATIVE EXAMPLE 2 4-(3-hydroxymethylphenylV2-mercaptothiazole

Step 1

Methyl 3-acetylbenzoate

A stirred solution of 3-acetylbenzoic acid (1.56 g, 9.5 mmol) in a 3: 1 solution of tetrahydrofuran - methanol (48 mL) was treated drop wise with a solution of trimethylsilyldiazomethane in hexane (2M, 5.0 mL, 10.0 mmol). After 15 min the solution was treated with glacial acetic acid (1.0 mL). After 15 min of stirring with acetic acid, the solution was diluted with ethyl acetate (50 mL) and washed with aqueous IM sodium hydroxide (40 mL), water (40 mL), IM hydrochloric acid (40 mL), water (40 mL), and brine. The organic phase was dried over magnesium sulfate, filtered, and evaporated under vacuum to give the title compound as a tan colored oil (1.10 g) which crystallized upon standing. lH NMR (CDCI3) δ 2.61 (s, COCH3), 3.91 (s, OCH3), 7.51 (t, J=7.8Hz, H-5), 7.49 - 8.21 (m, H-4, H-6), 8.55 (t, J=1.8Hz, H-2)

Step 2

Methyl 3-chloroacetylbenzoate

A stirred solution of methyl 3-acetylbenzoate (1.10 g. 6.17 mmol) in 3:2 glacial acetic acid - concentrated hydrochloric acid ( 10 mL) was bubbled with chlorine gas at room temperature. Gas bubbling was stopped after 5 min and the reaction flask was stoppered and stirred at room temperature for 5 min. The reaction mixture was then concentrated under vacuum to a white paste which was dissolved in diethyl ether (40

mL) and washed with water (40 mL), pH7 phosphate buffer (40 mL), brine, dried over magnesium sulfate, filtered and concentrated under vacuum to give the title compound as a white solid (1.25 g). - NMR (CDC13) δ 3.95 (s, OCH3), 4.73 (s, CH2C1), 7.58 (t, H-5), 8.1-8.8.3(m, H-4, H-6), 8.56 (t, H-2)

Step 3 2-Mercapto-4-(3-methoxycarbonylphenvπthiazole

A stirred aqueous suspension of ammonium dithiocarbamate (0.78 g, 7.06 mmol) was treated at room temeprature with a solution of methyl 3-chloroacetylbenzoate(1.25 g, 5.88 mmol) in tetrahydrofuran (8mL) and the mixture heated to 50°C in an oil bath. After 3 hr the reaction was removed from the bath and allowed to cool to room temperature. A two phase mixture resulted which was partitioned between ethyl acetate (25 mL) and water (25 mL). The organic phase was recovered, washed with brine, dried over magnesium sulfate, filtered and evaporated under vacuum to a semi-solid (1.49 g). The semi-solid product was mixed with dichloromethane (ca. 10 mL) and filtered to give the title compound as a white solid (321 mg).

-H NMR (DMSOdό) δ 3.38 (s, OCH3), 7.47 (s, thiazole H-5), 7.61 (t, J=7.8Hz, H-6), 7.97 (d, J=7.8Hz, Ar-H), 8.01 (d, J=7.8Hz, Ar=H), 8.34 (s, H-2), 13.84 (br s, NH)

13c NMR (DMSOdό) δ 52.76, 1 10.99, 126.87, 129.47, 129.87, 129.98, 130.79, 130.88, 140.96, 166.13, 190.16 MS (CI) (m/z) 252.1(M + l )

Step 4 2-Mercapto-4(3-hydroxymethylphenyDthiazole

A stirred solution of 2-mercapto-4-(3-methoxycarbonyl) thiazole (280 mg, 1.1 1 mmol) in tetrahydrofuran (5.4 mL) under a nitrogen atmosphere was treated with a solution of lithium aluminum hydride in tetrahydrofuran (I M, 1.1 mL, 1 ,1 mmol). The mixture was heated in an oil bath at 50°C and after 40 min the mixture was removed from the bath and allowed to cool to room temperature. The reaction was treated with 2N hydrochloric acid to give a solution of pH2 which was

extracted with ethyl acetate (10 mL). The extract was washed with brine, dried over magnesium sulfate, filtered, and evaporated under vacuum to a solid (237 mg). The crude product was flash silica gel column (2 x 15 cm) chromatographed eluting with 2:1 , ethyl acetate:hexane. Product containing fractions were combined and evaporated under vacuum to a semi-solid. The semi-solid was mixed with dichloromethane (10 mL) and filtered to afford the title compound (180 mg) as a white solid. -H NMR (DMSOdό) δ 4.51 (d, J=5Hz, CH20), 5.26 (T, J=5Hz, OH), 7.26 (s, thiazole H-5), 7.36 (t, J=7.5Hz, H-4), 7.39 (t, J=7.6Hz, H-5), 7.59 (d, J=7.6Hz, H-6), 7.67 (s, H-2)

13c NMR (DMSOd6) δ 63.01, 109.51, 124.32, 124.59, 127.60, 128.74, 129.12, 142.24, 143.78, 189.99 MS (CI) (m/z) 224.0(M + l)

PREPARATIVE EXAMPLE 3 4-phenyl-5-(hvdroxymethvD-2-mercapto-thiazole

Step l

4-phenyl-5-ethoxycarbonyl-2-bromothiazole A suspension of copper bromide (0.486g, 2.18mmol) in acetonitrile (lOmL) was cooled in an ice bath and treated with t-butyl nitrite (0.32mL, 2.7mmol). The mixture was stirred for 20 minutes whereupon 4-phenyl-5-ethoxycarbonyl-2-aminothiazole (0.45g, 1.81 mmol) was added and the suspension was removed from the ice bath.

After 15 minutes the mixture was partitioned between ethyl ether (40mL) and IN hydrochloric acid (30mL). The ether layer was washed with additional IN hydrochloric acid (30mL) and then brine (20mL). The ether layer was dried with magnesium sulfate, filtered and evaporated to give the title compound as an oil (500mg).

¹ H NMR (CDC1 , 500 MHz) δ 1.30 (t, CH ₂ CH ₃ ), 4.31 (q, CH2CH ₃ ).7.45

(m, 3ArH) and 7.76 (m, 2ArH).

¹³ C NMR (CDCI ₃ , 125 MHz) δ 14.0, 61.9, 127.8, 129.6, 129.9, 132.6,

139.8, 159.9 and 160.2.

Step 2

4-phenyl-5-(ethoxycarbonylV2-mercaptothiazole

Potassium hydrosulfide (0.4g, 5.5mmol) was added to a solution of 4-phenyl-5-(ethoxycarbonyl)-2-bromothiazole (0.5g, 1.6 mmol) in ethanol (6mL) and the mixture was stirred in a 80°C oil bath for

1 hour. After cooling to room temperature, the ethanol was evaporated and mixture was partitioned between ethyl acetate (50mL) and water

(30mL). The pH was made acidic and the ethyl acetate layer was washed with brine (20mL), dried with magnesium sulfate, filtered and evaporated to give the title compound as a light yellow solid (0.45g). NMR (DMSO-d ₆ , 500 MHz) δ 1.08 (t, CH2CH ₃ ), 4.07 (q, CH2CH ₃ ) and 7.4-7.6 (m, 5ArH).

¹³ C NMR (DMSO-dβ, 125 MHz) δ 14.2, 61.6, 1 13.6, 128.2, 128.2, 130.2,

130.7, 148.9, 159.0 and 189.4.

Step 3 4-phenyl-5-(hvdroxymethylV2-mercaptothiazole

A solution of 4-phenyl-5-(ethoxycarbonyl)-2-mercapto- thiazole (0.45g, 1.7mmol) in tetrahydrofuran (3mL) was treated with a tetrahydrofuran solution of lithium aluminum hydride (3.4 mL, 3.4 mmol). The mixture was stirred in a 60 °C oil bath for 10 minutes and after cooling to room temperature, was partitioned between ethyl acetate (20mL) and water ( lOmL). The pH was adjusted to 2 and the ethyl acetate layer was washed with brine ( lOmL), was dried with magnesium

sulfate, filtered and evaporated to give the title compound as a foam (400mg). ⁱ H NMR (2:1 CDC1 ₃ / CD ₃ OD, 500 MHz) δ 4.49 (s, CH ₂ OH) and 7.42 (m, 5 ArH). 13C NMR (2:1 CDCI ₃ / CD ₃ OD, 125 MHz) δ 55.9, 127.5, 128.1, 128.2, 128.8, 128.8, 129.2, 129.6, 138.3 and 188.4.

PREPARATIVE EXAMPLE 4 4-((5-hvdroxymethyl)thien-2-yl)-2-mercaptothiazole

Step 1

5-ethoxycarbonyl-2-(( 1 -trimethylsilyloxytethen- 1 -vPthiophene

A solution of 5-ethoxycarbonyl-2-acetyl-thiophene (0.2g, lmmol) in methylene chloride (3mL) was cooled in an ice bath under nitrogen. Triethylamine (0.188mL, 1.35mmol) was added, followed by the dropwise addition of trimethylsilyl trifluoromethanesulfonate (0.213mL, 1. lmmol) over 3 minutes. After 15 minutes, the mixture was partitioned between methylene chloride (lOmL) and 0.1N pH 7 phosphate buffer (lOmL). The methylene chloride layer was washed with additional buffer ( lxlOmL), aqueous sodium bicarbonate (lxlOmL) and was dried with magnesium sulfate, filtered and evaporated under vacuum to give a light yellow foam (265mg). Examination of the foam by HNMR showed it to be a 95:5 mixture of the title compound/ starting material. 'H NMR (CDC1 ₃ , 500 MHz) δ 0.29 (s, SiMe ₃ ), 1.38 (t, CH ₂ CH ₃ ), 4.34 (q, CH ₂ CH ₃₎ , 4.44 and 4.94 (s. 2CH), 7.15 and 7.65 (d, 2ArH).

¹³ C NMR (CDCI ₃ , 125 MHz) δ 0.06, 14.3, 61.1, 92.2, 123.8, 132.5, 133.5, 149.3, 150.2 and 162.3.

Step 2 5-ethoxycarbonyl-2-(2-bromoacetyl ^' )thiophene

A solution of 5-ethoxycarbonyl-2-((l-trimethylsilyloxy) ethen- 1 -yl)thiophene (0.265g, 0.98mmol) in anhydrous tetrahydrofuran (4mL) was cooled in an ice bath under nitrogen. N-bromosuccinimide (0.178g, lmmol) was added and the mixture was removed from the ice bath after 15minutes. After an additional 10 minutes, the mixture was partitioned between methylene chloride (lOmL) and aqueous sodium bicarbonate (20mL). The methylene chloride layer was dried with magnesium sulfate, filtered and evaporated under vacuum to give an oil (300mg, quantitative). Examination of the foam by HNMR showed it to be apprx. a 95:5 mixture of the title compound/ 5-ethoxycarbonyl- 2-acetyl-thiophene. ⁱ H NMR (CDCI ₃ , 500 MHz) δ 1.40 (t, CH2CH ₃ ), 4.37 (s, CH ₂ ), 4.39 (q, CH ₂ CH ₃₎ .7.75 and 7.79 (d, 2ArH). -- ^■ C NMR (CDCI ₃ , 125 MHz) δ 14.2, 30.3, 62.0, 132.8, 133.2, 141.0, 144.6, 161.3, and 184.6.

Step 3

4-((5-ethoxycarbonyDthien-2-yl)-2-mercaptothiazole

A solution of 5-ethoxycarbonyl-2-(2-bromoacetyl)thiophene (0.300g, lmmol) in tetrahydrofuran (4mL) was treated with a suspension of ammonium dithiocarbamate (0.136g, 1.23mmol) in water (2mL). The mixture was stiπed at room temperature for 4 hours and was then heated overnight in a 80°C oil bath. After cooling to room temperature, the mixture was partitioned between methylene chloride (lOmL) and water (6mL). The pH was adjusted to 4 and the aqueous layer was extracted with methylene chloride (lOmL). The combined extracts were dried with magnesium sulfate, filtered and evaporated to a foam. The foam was purified on preparative silica plates (EM Science, 2x1000, eluted with- 5% methanol in methylene chloride). The product band was removed.

eluted with 10% methanol in methylene chloride and evaporated to give the title compound (91mg). ^l H NMR (13:1 CDC1 ₃ / CD ₃ OD, 500 MHz) δ 1.34 (t, CH2CH ₃ ), 4.33 (q, CH ₂ CH ₃ ), 6.75 (s, ArH), 7.29 and 7.68 (d, 2ArH). ¹³ C NMR (13:1 CDCI ₃ / CD ₃ OD, 125 MHz) δ 14.1, 61.7, 110.0, 125.7, 133.8, 133.9, 135.3, 136.6, 161.8 and 190.2.

Step 4

4-((5-hvdroxymethvOthien-2-yD-2-mercaptothiazole A solution of 4-((5-ethoxycarbonyl)thien-2-yl)-2- mercaptothiazole (0.09 lg, 0.335mmol) in tetrahydrofuran (3mL) was treated with a tetrahydrofuran solution of lithium aluminum hydride (0.335mL, 0.335mmol). The mixture was stirred in a 80°C oil bath for 1 hour and after cooling to room temperature, was partitioned between ethyl acetate (20mL) and water (lOmL). The pH was adjusted to 2 and the ethyl acetate layer was washed with brine (lOmL), was dried with magnesium sulfate, filtered and evaporated to give the title compound as a foam (70mg).

^] H NMR (DMSO-d _ό , 500 MHz) δ 4.61 (s, CH ₂ OH), 5.60 (s, NH), 7.03 (s, ArH), 6.91 and 7.43 (d, 2ArH).

¹ C NMR (DMSO-d ₆ , 125 MHz) δ 58.7, 108.0, 124.8, 126.2, 129.8, 136.4, 148.5 and 189.8.

PREPARATIVE EXAMPLE 5 4-(5-(2-hvdroxyethvDthien-2-vD-2-mercaptothiazole

Ammonium dithiocarbamate

EtOH/ H ₂ 0

Step 1

Ethyl 2-(2-chloroacetyl thiophene-5-acetic acid

A suspension of 5-chloroacetylthiophene-2-acetic acid (0.2g, lmmol) in methylene chloride (3mL) was treated with oxalyl chloride (1.05mL, 12mmol) under nitrogen. Several drops of dimethylformamide were added and the mixture was stirred at room temperature for 1.5 hours. The solution was cooled in an ice bath and ice cold ethanol (lOmL) was added. After 0.5 hours the mixture was partitioned between methylene chloride (40mL) and saturated aqueous potassium carbonate (40mL). The aqueous layer was re-extracted with additional methylene chloride (lOmL) and the combined extracts were dried with magnesium sulfate, were filtered and evaporated to give the title compound as a dark oil (3g). -U NMR of the crude shows a 5: 1 mixture of the title compound/ diethyl oxalate. ⁱ H NMR (CDC1 ₃ , 500 MHz) δ 1.31 (t, CH ₂ CH ₃ ), 3.89 (s, CH ₂ C1), 4.23 (q, CH ₂ CH ₃₎ ,4.57 (s, CH ₂ Ar), 7.04 and 7.68 (d, 2ArH).

Step 2

4-(5-(ethoxycarbonylmethvDthien-2-vD-2-mercaptothiazole

A solution of ethyl 2-(2-chloroacetyl)thiophene-5-acetic acid (2.4g, 9.75mmol) in ethanol (20mL) was treated with ammonium dithiocarbamate (1.3g, 12mmol) and the mixture was stirred in a 80°C oil bath for 2 hours. After cooling to room temperature, the ethanol was evaporated and mixture was partitioned between ethyl acetate (50mL) and water (30mL). The ethyl acetate layer was washed with brine (20mL) and dried with magnesium sulfate, filtered and evaporated to an oil. The oil was extracted with ether (2x40mL) and gave 2,3g of an oil upon evaporation. The oil slowly solidified and was filtered with ethyl acetate (lOmL) to give the title compound as a light yellow solid (0.53g). iH NMR (13: 1 CDCI ₃ / CD ₃ OD, 500 MHz) δ 1.24 (t, CH ₂ CH ₃ ), 3.79 (s, CH ₂ Ar), 4.15 (q, Q CU^ 6.52 (s, ArH), 6.83 and 7.13 (d, 2ArH). 1 C NMR (13: 1 CDCI ₃ / CD ₃ OD, 125 MHz) δ 14.0, 35.4, 61.6, 107.4, 125.4, 127.7, 130.2, 136.0. 136.7. 170.2 and 189.8.

Step 3

4-((5-hydroxyethvDthien-2-ylV2-mercaptothiazole

A solution of 4-(5-(ethoxycarbonyl))-2-mercaptothiazole (0.4g, 1.4mmol) in tetrahydrofuran (2mL) was treated with a tetrahydro¬ furan solution of lithium aluminum hydride (2.8mL, 2.8mmol). The mixture was stirred in a 60°C oil bath for 1 hour and after cooling to room temperature, was partitioned between ethyl acetate (20mL) and water (lOmL). The pH was adjusted to 2 and the ethyl acetate layer was washed with brine (lOmL), dried with magnesium sulfate, filtered and evaporated to give the title compound as a foam (290mg). ⁱ H NMR (3:1 CDC1 ₃ / CD ₃ OD, 500 MHz) δ 2.99 (t, CH ₂ OH), 3.77 (t, CH ₂ Ar), 6.53 (s, ArH), 6.77 and 7.13 (d, 2ArH). ¹ C NMR (3:1 CDCI ₃ / CD ₃ OD, 125 MHz) δ 33.1 , 62.4, 107.0, 125.5, 126.0, 128.9, 136.6, 143.3, and 189.8.

PREPARATIVE EXAMPLE 6 4-(4-(2-hydroxyethyl)thien-2-vD-2-mercaptothiazole

Step 1

4-acetoxyethyl-2-acetylthiophene

Acetyl chloride ( 1.33mL. 18.73mmol) was added dropwise to ice cold 3-(2-hydroxyethyl)-thiophene ( I mL. 8.92mmol). After the exothermic reaction subsided, perchloric acid (0.05mL) was added.

Acetyl chloride (1.33mL, 18.73mmol) was added dropwise to ice cold 3-(2-hydroxyethyl)-thiophene (ImL, 8.92mmol). After the exothermic reaction subsided, perchloric acid (0.05mL) was added. The mixture was removed from the ice bath and stirred for 18 hours at room temperature. The dark solution was partitioned between methylene chloride (50mL) and saturated aqueous potassium carbonate (70mL). The methylene chloride layer was dried with magnesium sulfate, filtered and evaporated to an oil (1.93g). The oil was placed on a silica column (2x36cm EM science silica gel 60, eluted with 1 :1 ethyl ether/hexanes, collecting 8mL fractions) and fractions 8- 13 were combined to give 3-(2-acetoxyethyl)-2-acetylthiophene as a clear oil (550mg). Fractions 15-25 were combined to give the title compound as a clear oil (0.34g).

data for 3-(2-acetoxyethyl)-2-acetylthiophene; ⁱ H NMR (CDC1 ₃ , 500 MHz) δ 2.00 (s, OAc), 2.53 (s, Ac), 3.33 (t, ArCH ₂ ), 4.29 (t, CH ₂ OAc), 7.01 and 7.43 (d, 2ArH). ^!3 C NMR (CDCI ₃ , 125 MHz) δ 20.0, 29.4, 29.7, 63.7, 129.7, 131.8,

136.2, 145.0, 170.9 and 190.9. data for 4-(2-acetoxyethyl)-2-acetylthiophene; ⁱ H NMR (CDCI ₃ , 500 MHz) δ 2.05 (s, OAc), 2.53 (s, Ac), 2.95 (t, ArCH ₂ ), 4.28 (t, CH ₂ OAc), 7.34 and 7.56 (s, 2ArH). ¹ C NMR (CDCI ₃ , 125 MHz) δ 20.9, 26.8, 29.6, 63.8, 130.3, 133.3,

139.3, 144.4, 170.8 and 190.5.

Step 2

4-(4-(acetoxyethyl)thien-2-y0-2-mercaptothiazole

A solution of 4-(2-acetoxyethyl)-2-acetylthiophene (0.188g, 0.89mmol) in acetonitrile (5mL) was treated with hydroxy(tosyloxy) iodobenzene (HTIB) (0.52g, 1.33mmol) and the mixture was stirred in a 80°C oil bath for 2 hours. A suspension of ammonium dithiocarbamate (0.29g, 2.67mmol) in water (3mL) was added and the mixture was heated for an additional 18 hours. After cooling to room temperature, the mixture was partitioned between ethyl acetate (20mL) and water (lOmL). The pH was adjusted to 2 and the ethyl acetate layer was washed with

brine (20mL), dried with magnesium sulfate, filtered and evaporated to give a yellow solid. The solid was purified on preparative silica plates (3x1000, eluted with 10% methanol/ methylene chloride). The product band was removed, eluted with 10% methanol/ methylene chloride and was evaporated to give the title compound as a light yellow solid 0.163g). ⁱ H NMR (CDC1 ₃ 500 MHz) δ 2.05 (s, OAc), 2.54 (s, Ac), 2.95 (t, ArCH ₂ ), 4.28 (t, CH ₂ 0), 6.63 (s, ArH), 7.35 (s, ArH) and 7.56 (s, ArH).

Step 3 4-(4-(2-hydroxyethvDthien-2-vD-2-mercaptothiazole

IN sodium hydroxide (1.92mL, 1.92mmol) was added to a solution of 4-(4-(2-acetoxyethyl)thien-2-yl)-2-mercaptothiazole (0.163g, 0.77mmol) in ethanol (2mL). After 10 minutes, the mixture was partitioned between methylene chloride (8mL) and water (7mL). The pH was adjusted to 2 and the methylene chloride layer was dried with magnesium sulfate, filtered and evaporated to give the title compound as an oil (155mg). Examination of the oil by HNMR showed the presence of an equimolar amount of ethanol. »H NMR (CDCI ₃ , 500 MHz) δ 2.81 (t, ArCH ₂ ), 3.85 (t, CH ₂ 0), 6.51 (s, ArH), 7.00 (s, ArH) and 7.15 (s, ArH).

¹³ C NMR (CDCI ₃ , 125 MHz) δ 33.3, 62.2, 107.4, 122.8, 127.2, 130.4, 136.3, 140.3 and 189.5.

PREPARATIVE EXAMPLE 7

)-Nitrobenzyl ( 1 R.5R.6S)-2-(Trifluoromethylsulfonyl)oxy-6-r( 1 RV (triethylsil yloxy)ethvH- 1 -methylcarbapen-2-em-3-carbox ylate

A solution of (3S,4R)-4-[2(R)-4-diazo-4-((p-nitrobenzyl) oxy)carbonyl-3-oxo-but-2-yl]-3-[ l(R)-hydroxyethyl]-azetidin-2-one (2.00 g, 5.12 mmol) and rhodium(II) octanoate dimer (20 mg, 0.026 mmol) in anhydrous dichloromethane (20 mL) was heated at reflux and under a nitrogen atmosphere for 3.25 hours. After cooling to room temperature, the reaction mixture was placed in a dry ice-acetone bath and treated dropwise with triethylamine (0.786 mL, 5.64 mmol). The resulting solution was stirred -78°C and under a nitrogen atmosphere for 5 minutes, then treated dropwise with trifluoromethanesulfonic anhydride (0.904 mL, 5.38 mmol) and stirred at -78°C for an additional 35 minutes. More triethylamine ( 1.07 mL. 7.68 mmol) was added dropwise followed by the slow, dropwise addition of triethylsilyl trifluoromethanesulfonate ( 1.27 mL, 5.64 mmol). After stirring an additional 75 minutes at -78°C, the reaction mixture was removed from the cooling bath, diluted with dichloromethane (75 mL), and washed with water (3 x 100 mL). The organic phase was dried over magnesium sulfate, filtered and evaporated under vacuum to give an oil (3.34 g).

The crude product was purified by flash chromatography on a column of EM silica gel 60 (230-400 mesh, 3 x 19 cm, wet packed under dichloromethane). The column was eluted with dichloromethane, collecting ca. 15 mL fractions. Fractions 12-20 were combined and concentrated under vacuum to an oil that slowly deposited small, white crystalline islands on pumping under high vacuum. The oil-solid mixture (2.454 g) was treated with hexane (25 mL) and briefly solicited to give a copious crystalline precipitate. The mixture was diluted with more hexane (25 mL), then cooled in an ice bath for 20 minutes and filtered. The filter cake was washed with cold hexane (3 x 5 mL) and vacuum dried to provide the title compound (1.912 g) as a fluffy white solid. The filtrate and washings were concentrated under vacuum to ca. 10 mL, seeded, and stirred in an ice bath to provide additional product (0.1 7 g) as a white solid. IR (KBr) 2961, 2880, 1782, 1731 , 1521, 1438, 1341, 1290, 1217, 1 174, 1 144, 813, 738, and 609 cm- ⁱ . UV (dioxane) λ _ma χ 273 nm (ε 14,300).

-H NMR (CDC1 ₃ , 500 MHz) δ θ.61 (m, CH ₃ CH ₂ Si), 0.95 (t, CH ₃ CH ₂ Si), 1.25 (d, I -CH ₃ ), 1.30 (d, CH ₃ CHOH), 3.37 (dq, H-l), 3.40 (dd, H-6), 4.28 (p, CH ₃ CHOH), 4.35 (dd, H-5), 5.36 and 5.43 (two d's, CH ₂ C ₆ H N0 ₂ ), 7.63 and 8.23 (two m's, CH ₂ C ₆ H4N0 ₂ ).

EXAMPLE 1

SODIUM (5R.6S)-2-rTHIAZOL-2- YL1THIO-6-f(lR)-

HYDROXYETHYL1CARBAPEN-2-EM-3-CARBOXYLATE

H ₂ , 10% Pd/C

NaHC0 ₃

THF-EtOH-H ₂ 0

Step l p-Nitrobenzyl (5R.6SV2-rthiazol-2-vnthio-6-f( 1 R - hvdroxyethyllcarbapen-2-em-3-carboxylate

To 25 mg (0.21 mmol, 1.3 eq.) of 2-mercaptothiazole in 2 ml of anhydrous tetrahydrofuran at 0 °C was added 8.6 mg of 60% NaH oil dispersion (0.215 mmol). This mixture was stirred for 20 minutes until all of the sodium hydride was consumed, and a solution of 100 mg (0.172 mmol) of p-nitrobenzyl (5R,6S)-2-(diphenylphosphono)oxy-6- [(lR)-hydroxyethyl]carbapen-2-em-3-carboxylate in 2 ml of anhydrous tetrahydrofuran was added dropwise. The reaction was stiπed at room temperature for 45 minutes, after which time the solvent was removed under reduced pressure and the crude residue was chromatographed on EM silica gel 60 (230-400 mesh), eluting with 95 : 5 methylene chloride- methanol. In this manner 72 mg of the title compound was obtained as an oil. iH NMR (CDCI3, 400 MHz) δ 8.20 & 7.63 (two d's, CH2C6H4NO2), 7.89 & 7.50 (two d's, thiazole Ar-H), 5.50 & 5.26 (two d's,

CH2C6H4NO2), 4.25 (m, H-5 & CH3CHOH), 3.17 (dd, H-6), 2.92 (d, H-l), 1.28 (d, CH3CHOH).

Step 2

⁵ Sodium (5R.6S 2-rthiazol-2-yllthio-6-[( 1 R)-hvdroxyethyll carbapen-2-em-3-carboxylate

A solution was prepared of 56 mg (0.125 mmol) of p-nitro- benzyl (5R,6S)-2-[thiazol-2-yl]thio-6-[( 1 R)-hydroxyethyl]carbapen-2- em-3-carboxylate in 3.5 ml of tetrahydrofuran, 1.8 ml of ethanol, and 2.7 ι° ml of water containing 4.4 mg (0.052 mmol) of sodium bicarbonate. This solution was added to a vigorously stiπed, prehydrogenated mixture of 24 mg of 10% Pd/C in 1.8 ml of EtOH. This mixture was stirred briskly under a H2 atmosphere for 1 hour at room temperature then for 1 hour at 40 °C. The mixture was filtered through a Celite pad and the catalyst

¹⁵ washed with water (2 x 5 ml). The filtrate was washed with methylene chloride (2 x 5 ml) and ether (1 x 5 ml), centrifuging after each washing to break the emulsions, and then filtered through a 0.45 micron CR acrodisc, concentrated under vacuum to about 5 ml volume, and lyophilized to give the title compound (20 mg) as an amorphous white

²⁰ solid.

UV (H2O) λmax 300 nm (ε 5974). iH NMR (D20, 400 MHz) δ 7.90 & 7.77 (two d's, thiazole Ar-H), 4.17 (m, H-5 & CH3CHOH), 3.38 (dd, H-6), 2.90 & 2.70 (two dd's, H's-1 ), 1.21 (d, CH3CHOH).

²⁵ MS, m/z 357 (M+Na).

30

EXAMPLE 2

SODIUM ( lR.5S.6SV2-r(2-THIAZOLYLyTHI01-

6-\( 1 R)HYDROX YETHYL1- 1 -METHYLCARB APEN-

2-EM-3-CARBOXYLATE

H ₂ , 10% Pd/C

NaHC0 ₃ THF-EtOH-H ₂ 0

Step 1 p-Nitrobenzyl (lR.5S.6SV2-[(2-thiazolvnthiol-6-[(lR)- hydroxyethyll-l-methylcarbapen-2-em-3-carboxylate

To 25 mg (0.21 mmol, 1.3 eq.) of 2-mercaptothiazole in 2 ml of anhydrous tetrahydrofuran at 0 °C was added 8.4 mg of 60% NaH oil dispersion (0.21 mmol). This mixture was stirred for 20 minutes ₅ until all of the sodium hydride was consumed, and a solution of 100 mg (0.172 mmol) of p-nitrobenzyl (lR,5R,6S)-2-(diphenylphosphono)oxy- 6-[(lR)-hydroxyethyl]-l-methylcarbapen-2-em-3-carboxylate in 2 ml of anhydrous tetrahydrofuran was added dropwise. The reaction was stirred at room temperature for 2 hours, after which time the solvent was o removed under reduced pressure and the crude residue was chroma- tographed on EM silica gel 60 (230-400 mesh), eluting with 4 : 1 methylene chloride -ethyl acetate. In this manner 61 mg of the title compound was obtained as an oil. i H NMR (CDC13, 400 MHz) δ 8.20 & 7.64 (two d's. CH2C6H4NO2), 7.85 & 7.48 (two d's. thiazole Ar-H). 5.50 & 5.26 (two d's.

CH2C6H4NO2), 4.26 (dd, H-5), 4.22 (p, CH3CHOH), 3.48 (dq, H-l), 3.26 (dd, H-6), 1.29 (d, CH3CHOH), 1.05 (d, I-CH3).

Step 2 Sodium (lR.5S.6SV2- (2-thiazolvnthiol-6-[(lRVhvdroxyethyll-l- methylcarbapen-2-em-3-carboxylate

A solution was prepared of 31 mg (0.052 mmol) of p-nitro- benzyl (5S,6S)-2-[thiazol-2-yl]thio-6-[(lR)-hydroxyethyl]-l -methyl- carbapen-2-em-3-carboxylate in 3.5 ml of tetrahydrofuran, 1.8 ml of ethanol, and 2.7 ml of water containing 4.4 mg (0.052 mmol) of sodium bicarbonate. This solution was added to a vigorously stirred, prehydro- genated mixture of 24 mg of 10% Pd/C in 1.8 ml of EtOH. This mixture was stiπed briskly under a H2 atmosphere for 1 hour at room temperature (an additional 24 mg of 10% Pd/C was added after 30 minutes) then for 1 hour at 40 °C. The mixture was filtered through a Celite pad and the catalyst washed with water (2 x 5 ml). The filtrate was washed with methylene chloride (2 x 5 ml) and ether (1 x 5 ml), centrifuging after each washing to break the emulsions, and then filtered through a 0.45 micron CR acrodisc, concentrated under vacuum to about 5 ml volume, and lyophilized to give the title compound (1 1 mg) as an amorphous white solid.

UV (0.05M pH 7.0 MOPS buffer) λ, _ntιx 303 nm (ε 6273). iH NMR (D20, 400 MHz) δ 7.86 & 7.72 (two d's, thiazole Ar-H), 4.22 (m, H-5 & CH3CHOH), 3.45 (dd, H-6), 3.19 (dq, H-l ), 1.25 (d, CH3CHOH), 1.06 (d, 1 -CH3).

EXAMPLE 3

SODIUM (5R.6SV2-r2-THIENYLlTHIO-6-rf lRV

HYDROXYETHYL1CARBAPEN-2-EM-3-CARBOXYLATE

H ₂ , 10% Pd/C

NaHC0 ₃

THF-EtOH-H ₂ 0

Step 1 p-Nitrobenzyl (5R.6S)-2-[2-thienvnthio-6-[( 1 RVhvdroxyethyllcarbapen- 2-em-3-carboxylate

To 25 mg (0.21 mmol, 1.3 eq.) of 2-mercaptothiophene in 2 ml of anhydrous tetrahydrofuran at 0 °C was added 8.6 mg of 60% NaH oil dispersion (0.215 mmol). This mixture was stiπed for 10 minutes until all of the sodium hydride was consumed, and a solution of 100 mg (0.172 mmol) of p-nitrobenzyl (5R,6S)-2-(diphenylphosphono)oxy-6- [(lR)-hydroxyethyl]carbapen-2-em-3-carboxylate in 2 ml of anhydrous tetrahydrofuran was added dropwise. The reaction was stirred at 0°C for 45 minutes, after which time the solvent was removed under reduced pressure and the crude residue was chromatographed on EM silica gel 60 (230-400 mesh), eluting with 95 : 5 methylene chloride-methanol. In this manner 49 mg of the title compound was obtained as an oil. i H NMR (CDC13, 400 MHz) δ 8.20 & 7.64 (two d's, CH2C6H4NO2), 7.49 (d, thiophene Ar-H), 7.25 (d, thiophene Ar-H), 7.04 (dd, thiophene Ar-H), 5.50 & 5.27 (two d's, CH2C6H4NO2), 4.17 (br p, CH3CHOH), 4.1 1 (ddd, H-5). 3.13 (dd, H-6), 2.80 (d. H- 1 ), 1.28 (d, CH3CHOH).

Step 2

Sodium (5R.6S -2-12-thienyllthio-6-[(l R)-hvdroxyethyllcarbapen-2-em- 3-carboxylate

A solution was prepared of 49 mg (0.11 mmol) of p-nitrobenzyl (5R,6S)- 2-[2-thienyl]thio-6-[(lR)-hydroxyethyl]carbapen-2-em-3-carbo xylate in 7 ml of tetrahydrofuran, 3.6 ml of ethanol, and 2.75 ml of water containing 8.8 mg (0.104 mmol) of sodium bicarbonate. This solution was added to a vigorously stiπed, prehydrogenated mixture of 48 mg of 10% Pd/C in 3.5 ml of EtOH. This mixture was stirred briskly under a H2 atmosphere for 1 hour at room temperature. An additional 25 mg of 10% Pd/C was added and the resulting mixture was stiπed at 40 °C for 30 minutes. The mixture was filtered through a Celite pad and the catalyst washed with water (2 x 5 ml). The filtrate was washed with methylene chloride (2 x 5 ml) and ether (1 x 5 ml), centrifuging after each washing to break the emulsions, and then filtered through a 0.45 micron CR acrodisc, concen¬ trated under vacuum to about 5 ml volume, and lyophilized to give the title compound (20 mg) as an amorphous white solid. UV (H20) λmax 297 nm (ε 9598). H NMR (D20, 400 MHz) δ 7.64 (d, thiophene Ar-H), 7.36 (d, thiophene Ar-H), 7.12 (dd, thiophene Ar-H), 4.18 (p, CH3CHOH), 4.08 (ddd, H-5), 3.20 (dd, H-6), 2.89 & 2.70 (two dd's, H's-1 ), 1.21 (d, CH3CHOH).

MS, m/z 334 (M+H), 356 (M+Na).

EXAMPLE 4

SODIUM (lR.5S.6S -2-[(2-THIENYL THI01-6-[(l R)-

HYDROXYETHYL1-1-METHYLCARBAPEN-2-EM-3-

CARBOXYLATE

H ₂ , 10% Pd/C

NaHC0 ₃

THF-EtOH-H ₂ 0

Step 1 p-Nitrobenzyl (lR.5S.6SV2-[(2-thienvnthio1-6-f(lR)-hvdroxyethvn-l - methylcarbapen-2-em-3-carboxylate

To 25 mg (0.21 mmol, 1.3 eq.) of 2-mercaptothiophene in 2 ml of anhydrous tetrahydrofuran at 0 °C was added 6.9 mg of 60% NaH oil dispersion (0.17 mmol). This mixture was stiπed for 1 hour then a solution of 100 mg (0.172 mmol) of p-nitrobenzyl (1R,5R.6S)- 2-(diphenylphosphono)oxy-6-[( 1 R)-hydroxyethyl]- 1 -methylcarbapen- 2-em-3-carboxylate in 2 ml of anhydrous tetrahydrofuran was added dropwise. The reaction was stiπed at room temperature for 15 minutes, after which time the solvent was removed under reduced pressure and the crude residue was chromatographed on EM silica gel 60 (230-400 mesh), eluting with 6 : 1 methylene chloride-ethyl acetate. In this manner 66 mg of the title compound was obtained as an oil. iH NMR (CDC13, 400 MHz) δ 8.21 & 7.67 (two d's. CH2C6H4NO2), 7.50 (d, thiophene Ar-H), 7.29 (d, thiophene Ar-H), 7.05 (t, thiophene Ar-H). 5.52 & 5.27 (two d's. CH2C6H4NO2), 4.20 (p. CH3CHOH), 4.16

(dd, H-5), 3.20 (dd, H-6), 3.10 (dq, H-l ), 1.30 (d, CH3CHOH), 1.09 (d, I-CH3).

Step 2 Sodium (lR.5S.6SV2-[(2-thienvnthio1-6-[(lR -hvdroxyethyll- l-methylcarbapen-2-em-3-carboxylate

A solution was prepared of 55 mg (0.12 mmol) of p-nitro- benzyl (5S,6S)-2-[2-thieny l]thio-6-[( 1 R)-hydroxyethyl]- 1 -methyl- carbapen-2-em -3 -carboxylate in 7 ml of tetrahydrofuran, 3.6 ml of ethanol, and 2.75 ml of water containing 8.8 mg (0.104 mmol) of sodium bicarbonate. This solution was added to a vigorously stiπed, prehydro- genated mixture of 48 mg of 10% Pd/C in 3.5 ml of EtOH. This mixture was stiπed briskly under a H2 atmosphere for 1 hour at 4 °C. The mixture was filtered through a Celite pad and the catalyst washed with water (2 x 5 ml). The filtrate was washed with methylene chloride (2 x 5 ml) and ether (1 x 5 ml), centrifuging after each washing to break the emulsions, and then filtered through a 0.45 micron CR acrodisc, concentrated under vacuum to about 5 ml volume, and lyophilized to give the title compound (25 mg) as an amorphous white solid. UV (0.05M pH 7.0 MOPS buffer) λ _ma * 305 nm (ε 1 1299). iH NMR (D20, 400 MHz) δ 7.69 (d, thiophene Ar-H), 7.42 (d, thiophene Ar-H), 7.16 (t, thiophene Ar-H), 4.22 (p, CH3CHOH), 4.12 (dd, H-5), 3.39 (dd, H-6), 3.06 (dq, H-l), 1.27 (d, CH3CHOH), 1.10 (d, 1-CH3).

EXAMPLE 5

SODIUM (5R.6S -2-r4-PHENYLTHIAZOL-2-YL1THIO-6-r(lRV

HYDROXYETHYL1CARBAPEN-2-EM-3-CARBOXYLATE

H ₂ , 10% Pd/C

NaHC0 ₃

THF-EtOH-H ₂ 0

Step l p-Nitrobenzyl (5R.6S -2-r4-phenylthiazol-2-vnthio-6-[(l R)- hvdroxyethyllcarbapen-2-em-3-carboxylate

To 42 mg (0.21 mmol, 1.3 eq.) of 2-mercapto-4-phenyl- thiazole in 2 ml of anhydrous tetrahydrofuran at 0 °C was added 6.9 mg of 60% NaH oil dispersion (0.17 mmol). This mixture was stirred for 30 minutes, then a solution of 100 mg (0.172 mmol) of p-nitrobenzyl (5R,6S)-2-(diphenylphosphono)oxy-6-[(lR)-hydroxyethyl]carbap en- 2-em-3-carboxylate in 2 ml of anhydrous tetrahydrofuran was added dropwise. The reaction was stiπed at room temperature for 10 minutes, after which time the solvent was removed under reduced pressure and the crude residue was chromatographed on EM silica gel 60 (230-400 mesh), eluting with 7: 1 methylene chloride-ethyl acetate. In this manner 40 mg of the title compound was obtained as an oil. i H NMR (CDC13, 400 MHz) δ 8.21 & 7.64 (two d's, CH2C6H4NO2), 7.88 (d, o-Ph-H), 7.61 (s, thiazole Ar-H), 7.43 (t, m-Ph-H), 7.37 (t, p-Ph-

H), 5.52 & 5.29 (two d's, CH2C6H4NO2), 4.19 (m, H-5 & CH3CHOH), 3.18 (dd, H-6), 3.09 & 2.99 (two dd's, H-l ), 1.29 (d, CH3CHOH).

Step 2 Sodium (5R.6SV2-[4-phenylthiazol-2-yllthio-6-r(lR)-hvdroxyethyll carbapen-2-em-3-carboxylate

A solution was prepared of 40 mg (0.076 mmol) of p-nitro- benzyl (5R,6S)-2-[4-phenylthiazol-2-yl]thio-6-[(lR)-hydroxyethyl] carbapen-2-em-3-carboxylate in 4.8 ml of tetrahydrofuran, 2.5 ml of ethanol, and 1.9 ml of water containing 6.1 mg (0.072 mmol) of sodium bicarbonate. This solution was added to a vigorously stirred, prehydro- genated mixture of 33 mg of 10% Pd/C in 2.5 ml of EtOH. This mixture was stirred briskly under a H2 atmosphere for 15 minutes at room temperature then for 15 minutes at 40 °C. The mixture was filtered through a Celite pad and the catalyst washed with water (2 x 5 ml). The filtrate was washed with methylene chloride (2 x 5 ml) and ether (1 x 5 ml), centrifuging after each washing to break the emulsions, and then filtered through a 0.45 micron CR acrodisc, concentrated under vacuum to about 5 ml volume, and lyophilized to give the title compound (8 mg) as an amorphous white solid.

UV (H20) λmax 292 nm (ε 8100). iH NMR (D20, 400 MHz) δ 7.95 (s, thiazole Ar-H), 7.88 (d, o-Ph-H), 7.55 (t, m-Ph-H), 7.48 (t, p-Ph-H), 4.18 (m, H-5 & CH3CHOH), 3.29 (dd, H-6), 2.98 & 2.84 (two dd's, H's-1 ). 1.22 (d, CH3CHOH).

EXAMPLE 6

SODIUM (1R.5S.6S 2-[(4-PHENYL-2-THIAZOLYL THI01-6-[( 1 RV

HYDROXYETHYL1-1-METHYLCARBAPEN-2-EM-3-

CARBOXYLATE

H ₂ , 10% Pd/C

NaHC0 ₃

THF-EtOH-H ₂ 0

Step 1 p-Nitrobenzyl (lR.5S.6S -2-l(4-phenyl-2-thiazolvnthiol-6-r(lRV hydroxyethyll-l -methylcarbapen-2-em-3-carboxylate

To 42 mg (0.21 mmol, 1.3 eq.) of 2-mercapto-4-phenyl- thiazole in 2 ml of anhydrous tetrahydrofuran at 0 °C was added 6.8 mg of 60% NaH oil dispersion (0.17 mmol). This mixture was stirred for 20 minutes then a solution of 100 mg (0.172 mmol) of p-nitrobenzyl ( 1 R,5R,6S)-2-(diphenylphosphono)oxy-6-[ ( 1 R)-hydroxyethy 1] - 1 -methy 1- carbapen-2-em-3-carboxylate in 2 ml of anhydrous tetrahydrofuran was added dropwise. The reaction was stiπed at room temperature for 25 minutes, after which time the solvent was removed under reduced pressure and the crude residue was chromatographed on EM silica gel 60 (230-400 mesh), eluting with 7: 1 methylene chloride-ethyl acetate. In this manner 59 mg of the title compound was obtained as an oil. iH NMR (CDC13. 400 MHz) δ 8.21 & 7.67 (two d's, CH2C6H4NO2), 7.88 (d, o-Ph-H). 7.57 (s, thiazole Ar-H), 7.43 (t, m-Ph-H), 7.37 (t. p-Ph-

H), 5.52 & 5.28 (two d's, CH2C6H4NO2). 4.29 (dd, H-5), 4.23 (p, CH3CHOH), 3.62 (dq, H-l), 3.18 (dd, H-6), 1.30 (d, CH3CHOH), 1.1 1 (d, 1-CH3). Step 2

Sodium ( 1 R.5S.6S V2-lT2-thiazol vnthio1-6-[( 1 RVhvdroxyethyll- 1 - methylcarbapen-2-em-3-carboxylate

A solution was prepared of 59 mg (0.052 mmol) of p-nitro¬ benzyl (5S,6S)-2-[thiazol-2-yl]thio-6-[( 1 R)-hydroxyethyl]- 1 -methyl- carbapen-2-em-3-carboxylate in 7 ml of tetrahydrofuran, 3.6 ml of ethanol, and .2.75 ml of water containing 8.8 mg (0.104 mmol) of sodium bicarbonate. This solution was added to a vigorously stirred, prehydro- genated mixture of 48 mg of 10% Pd/C in 3.5 ml of EtOH. This mixture was stirred briskly under a H2 atmosphere for 1 hour at room temperature (an additional 40 mg of 10% Pd/C was added after 40 minutes). The mixture was filtered through a Celite pad and the catalyst washed with water (2 x 5 ml). The filtrate was washed with methylene chloride (2 x 5 ml) and ether (1 x 5 ml), centrifuging after each washing to break the emulsions, and then filtered through a 0.45 micron CR acrodisc, concentrated under vacuum to about 5 ml volume, and lyophilized to give the title compound (25 mg) as an amorphous white solid. UV (0.05M pH 7.0 MOPS buffer) λ _max 313 nm (ε 9867). H NMR (D20, 400 MHz) δ 7.89 (s, thiazole Ar-H), 7.82 (d, o-Ph-H), 7.52 (t, m-Ph-H), 7.46 (t, p-Ph-H), 4.22 (m, H-5 & CH3CHOH), 3.36 (dd, H-6), 3.30 (dq, H's-1), 1.24 (d, CH3CHOH), 1.06 (d, I -CH3). MS, m/z 425 (M+H), 447 (M+Na).

EXAMPLE7

SODIUM(1R.5S.6SV21(5-PHENYLTHIAZOL-2-YL)THI01-6-(lR-

HYDROXYETHYD-1-METHYLCARBAPEN-2-EM-3-

CARBOXYLATE

Step l

4-Nitrobenzyl (1R.5S. 6S)-2[(5-phenylthiazol-2-vnthiol-6-(lR- triethylsil yloxyeth- 1 -vP- 1 -methylcarbapen-2-em-3-carboxylate A stiπed solution of 2-mercapto-5-phenyl-thiazole (67 mg, 0.345 mmol) in dry tetrahydrofuran (3.4 mL) under a nitrogen atmosphere was cooled in an ice bath and the cold solution treated with a tetrahydrofuran solution of lithium bis(trimethylsilyl)amide (IM, 0.33 mL, 0.33 mmol). After 25 min the solution was treated with 4-nitro- benzyl (1R,5R, 6S)-2-trifluoromethanesulfonyloxy)-6-(lR-triethylsilyl- oxyeth-l-yl)-l -methylcarbapen-2-em-3-carboxylate (220 mg, 0.329 mmol) and the solution was removed from the ice bath to stir at room temeprature. After 70 min the reaction was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate (5 mL), water (5 mL), dried over magnesium sulfate, filtered, and concentrated under vacuum to an oil and lyophilized from benzene to give (237 mg). The crude product was flash silica gel column (2 x 18 cm) chromatographed

eluting with 1 :4, ethyl acetate :hexane. Product containing fractions were combined, evaporated under vacuum, and lyophilized from benzene to give the title compound as an amorphous white solid (101 mg). 1H NMR (CDC13) δ 0.58 (q, J=8Hz, S1CH2CH3), 0.92(t, J=8Hz, S1CH2CH3), 1.12(d, J=7.3Hz, I-CH3), 1.22(d, J=6.2Hz, C//3CHOH), 3.27(dd, J=2.9, 5.5Hz, H-6), 3.66(dq, J=7.3, 9.9Hz, H-l ), 4.26(~p, J=~6Hz, CH3C /OH), 4.31(dd, J=2.9, 9.8Hz, H-5), 5.31(d, J=13.8Hz, C02C// _α Hb), 5.50(d, J=13.8Hz, Cθ2CH _a b), 7.38-7.56 (m, Ar H), 7.58(s, thiazole H5), 7.68(d, J=8.8Hz, ArH), 8.23(d, J=8.8Hz, ArH) IR(KBr)3433,2956,2876, 1779, 1708, 1523, 1340cm"l MS(m/z)537.9(M+)

Step2

4-Nitrobenzyl (1R.5S. 6S -2r(5-phenylthiazol-2-vnthiol-6-(lR- hvdroxyethyP- 1 -methylcarbapen-2-em-3-carboxylate

A stiπed ice cold solution of 4-nitrobenzyl (1R,5S, 6S)- 2[(5-phenylthiazol-2-yl)thio]-6-( 1 R-triethylsilyloxyeth- 1 -y 1)- 1 -methy 1- carbapen-2-em-3-carboxylate (90 mg, 0.138 mmol) in tetrahydrofuran (2.0 mL) under a nitrogen atmosphere was treated with glacial acetic acid (0.47 mL, 0.828 mmol) and a solution of tetrabutylammonium fluoride in tetrahydrofuran (IM, 0.275 mL, 0.275 mmol). After 2 hr additional tetrabutylammonium fluoride solution (IM, 0.275 mL, 0.275 mmol) added. After 2.75 hr the reaction was diluted with ethyl acetate (-10 mL), and washed with water (2 x 5 mL), brine, dried over magnesium sulfate, filtered, and evaporated under vacuum to an oil (91 mg). The crude product was flash silica gel column (2 x 1 1 cm) chromatographed eluting with 4:6, ethyl acetate :hexane. Product containing fractions were combined, evaporated under vacuum, and lyophilized from benzene to give the title compound (66 mg). 1H NMR (CDC13) δ 1.14(d, J=7.3Hz, I -CH3), 1.32(d. J=6.2Hz,

C/ 3CHOH), 3.30(dd, J=2.9, 6.6Hz, H-6), 3.64(dq, J=7.3, 9.9Hz, H-l ), 4.24(~p, J=~6Hz, CH3C OH), 4.30(dd, J=2.9, 9.8Hz, H-5), 5.30(d, J=13.5Hz, C02C//øHb), 5.53(d, J=13.5Hz, Cθ2CH _a Hb\ 7.36-7.46(m, Ar H), 7.58(s, thiazole H5), 7.67(d J=8.5Hz. ArH), 7.9 l (d, J=8Hz, Ar H), 8.23(d. J=8.5Hz, ArH)

Step 3

Sodium (lR.5S. 6SV2[(5-phenylthiazol-2-vnthio1-6-(lR-hvdroxyethvn- l-methylcarbapen-2-em-3-carboxylate

A solution of 4-nitrobenzyl (1R,5S, 6S)-2[(5-phenylthiazol-2-yl)thio]-6-

5 ( 1 R-hydroxyethyl)- 1 -methylcarbapen-2-em-3-carboxylate (59 mg, 0.1 1 mmol) in tetrahydrofuran (7.5 mL), was mixed with a solution of sodium bicarbonate (9.2 mg, 0.1 1 mmol) in water (5.6 mL), and ethanol added (7.5 mL) followed by 10% palladium on carbon (59 mg). The mixture was purged with nitrogen and hydrogen and hydrogenated under a l o balloon of hydrogen at room temperature. After 70 min the reaction was nitrogen purged and the mixture filtered. The filtrate was washed with dichloromethane (2 x 50 mL) followed by diethyl ether (2 x 50 mL). The aqueous phase was concentrated under vacuum, filtered through a 0.45mm filter and lyophilized to give the title compound as an off-white

15 fluffy solid (38 mg). lH NMR (D2θ) δ 1.07(d, J=7.0Hz, I-CH3), 1.23(d, J=6.3Hz, O/jCHOH), 3.30(m H-l,) 3.45(dd, J=2.5 5.9Hz, H-6), 4.18-4.24(m, H- 5, H-8), 7.43-7.82(m, Ar H), 7.51 (s, thiazole H5), 7.82-9.17 (m, Ar H) IR(KBr) 3424 1754, 1601, 1397 cπr*

20 UV(0.1 M pH7 MOPS buffer) λmax 315 εl 0,500

25

30

EXAMPLE 8

( 1 R.5S.6S -2f 2- ( 3- ( 4-[CARB AMOYLMETHYL- 1.4-

DIAZOBICYCLθr2.2.21QCT-l-YL)METHYLlPHENYL)THIAZOL-2-

YL ) THIQ1-6-( 1 R-HYDROXYETHYL 1 -METHYLCARB APEN-2-

EM-3-CARBOXYLATE CHLORIDE

R = OH

MsCI. Et ₃ N /CH2CI2

Tf = SQ ₂ CF ₃ & Ms = SQ ₂ C H ₃ | R = OMs Z2 Nal / acetone

R =

Step l

4-Nitrobenzyl (1 R.5S. 6S)-2r(4-r4-hvdroxymethylphenvnthiazol-2- vπthio1-6-( 1 R-triethylsilyloxyeth- 1 -yl)- 1 -methylcarbapen-2-em-3- carboxylate

A solution of 4-(4-hydroxymethylphenyl)-2-mercapto- thiazole (81 mg, 0.36 mmol) in dry tetrahydrofuran (3.4 mL) at room temperature was treated with powdered lithium hydroxide monohydrate (18 mg, 0.43 mmol) to give a light yellow solution. The solution was treated with 4-nitrobenzyl (1R,5R. 6S)-2-trifluoromethanesuIfonyloxy)-

6-(lR-triethylsilyloxyeth-l-yl)-l-methylcarbapen-2-em-3 -carboxylate (220 mg, 0.36 mmol), stiπed 105 min, diluted with ethyl acetate (15 mL), and washed with water (2 x 10 mL), brine, dried over magnesium sulfate, filtered, and evaporated under vacuum to a foam (303 mg). The crude product was flash silica gel column (2 x 14 cm) chromatographed eluting with 1 : 1, ethyl acetate:hexane. Product containing fractions were combined, evaporated under vacuum, and lyophilized from benzene to give the title compound as an amorphous solid (1 5 mg). 1H NMR (CDC13) δ 0.59 (q, J=8.0Hz, S1CH2CH3), 0.93(t, J=8.0Hz, SJC//2CH3), 1.12(d, J=7.4Hz, I-CH3), 1.22(d, J=6.2Hz, C/ 5CHOH), 3.27(dd, J=3.0, 5.5Hz, H-6), 3.65(dq, J=7.4, 9.9Hz, H-l ), 4.26(p, J=6Hz, CH3C /OH), 4.31(dd, J=3.0, 9.9Hz, H-5), 4.75(s, Ar-CH2), 5.31 (d, J=14.0Hz, C02C / _α Hb), 5.50(d, J=14.0Hz, Cθ2CH _a / ), 7.44 (d, J=8.2Hz, ArH), 7.57(s, thiazole H5), 7.67(d, J=8.7Hz, ArH), 7.89 (d, J=8.2Hz, ArH), 8.22(d, J=8.7Hz, ArH)

IR(KBr) 3449, 2957,2876, 1777, 1523, 1340cm-1 MS m/z682.2(M+ 1)

Step2 4-Nitrobenzyl ( 1 R.5S.6S1-2-f(4-(4-methanesulfonyloxymethylphenvπ- thiazol-2vπthio1-6-(lR-triethylsilyloxyeth-l -yl )-l -methylcarbapen-2-em- 3-carboxylate

A solution of 4-nitrobenzyl (lR,5S,6S)-2[(4-(4-hydroxymethylphenyl) thiazol-2-yl)thio]-6-(lR-triethylsilyloxyeth-l -yl)-l-methylcarbapen-2- em-3-carboxylate (182 mg, 0.273 mmol) in dry dichloromethane (3 mL) under a nitrogen atmosphere was cooled in an ice bath. The cold solution was treated with triethylamine (0.57 mL, 0.410 mmol) follwed by methanesulfonyl chloride (0.27 mL, 0.343 mmol). After 30 min the solution was diluted with dichloromethane (7 mL) and washed with water (5 mL), 0.5M HC1 (5 mL), and water (2 x 5 mL), dried over magnesium sulfate, filtered, and evaporated to afford the title compound as an oil (196 mg).

1H NMR (CDC13) δ 0.59 (q, J=7.8Hz, SiCH2CHjj), 0.92(t, J=7.8Hz, SiCH2CH3), 1.12(d, J=7.4Hz, I -CH3), 1.22(d, J=6.2Hz, CH3CHOH), 2.96(s, CH3SO2), 3.28(dd, J=2.9. 5.4Hz, NCH), 3.64(dq, J=7.4, 10.0Hz,

H- l), 4.26(~p, J=~6Hz, CH3C OH), 4.32(dd, J=2.9, 10.0Hz, H-5), 5.28(s, Ar-CH2), 5.3 l(d, J=13.8Hz, Cθ2C _α Hb), 5.50(d, J=13.8Hz, C02CH _a Hb), 7.50(d, J=8.4Hz, ArH), 7.62(s, thiazole H5), 7.68(d, J=8.8Hz, ArH), 7.93(d, J=8.3Hz, ArH), 8.23(d, J=8.8Hz, ArH) IR(KBr) 3450, 2957, 1777, 1708, 1522, 1340 cm-1 MS m/z 614.1 (M-OSiEt3 + 2)

Step 3 4-Nitrobenzyl (lR5S.6SV2-[4-(4-iodomethylphenvnthiazol-2-vnthiol-6- (lR-triethylsilyloxyeth-l-vπ-l-methylcarbapen-2-em-3-carbox ylate

A stiπed solution of 4-nitrobenzyl (lR,5S,6S)-2-[(4-(4-methanesulfonyl- oxymethylphenyl)thiazol-2yl)thio]-6-(lR-triethylsilyloxyeth- l-yl)-l - methylcarbapen-2-em-3-carboxylate (190 mg, 0.250 mmol) in acetone (3 mL) was treated with sodium iodide (112 mg, 0.750 mmol) and stiπed at room temperature. After 30 min the reaction was diluted with ethyl acetate (15 mL) and washed with water (2 x 15 mL), brine, dried over magnesium sulfate, filtered, and evaporated under vacuum to an oil (206 mg).

The crude product was flash silica gel column (2 x 5 cm) chromatographed eluting with 2:1, hexane :ethyl acetate. Product containing fractions were combined and evaporated under vacuum to a partial solid and lyophilized from benzene to afford the title compound (184 mg) as a white solid. iH NMR (CDCI3) δ 0.59 (q, J=7.8Hz, S1CH2CH3), 0.93(t, J=7.8Hz, SiO/2CH3), 1.1 l(d J=7.3Hz, I-CH3), 1.22(d, J=6.2Hz, C//3CHOH), 3.26(dd, J=2.9, 5.4Hz, H-6), 3.64(dq, J=7.3, 10.0Hz, H-l) , 4.26(p, J=6Hz, CH3C OH), 4.31 (dd, J=2.9, 10.0Hz, H-5), 4.50(s, ArCH2), 5.31(d, J=13.9Hz, Cθ2C / _fl Hb), 5.50(d, J=13.9Hz, Cθ2CH _a tf/>), 7.45 (d, J=8.2Hz, ArH), 7.58(s, thiazole H5). 7.68(d, J=8.8Hz, ArH), 7.82 (d, J=8.2Hz, ArH), 8.23(d, J=8.8Hz, ArH) IR(KBr) 3449, 2956, 1778, 1522, 1340 cm-1 MS m/z 792.0 (M+l )

Step 4 4-Nitrobenzyl ( 1 R.5S.6S 2-1-3 ( -4-[carbamoylmethyl- 1.4- diazobicyclor2.2.21oct- 1 -yPmethyllphen yl ) thiazol-2-yl I thiol-6( 1 R-triethylsilyloxyeth- 1 -yP- 1 -methylcarbapen- 2-em-3-carboxylate bis trifluoromethylsulfonate

A stiπed solution of 4 nitrobenzyl-(lR,5S,6S)-2-[4-(4- iodomethylphenyl)thiazol-2-yl)thio]-6-(l R-triethylsilyloxyeth- 1-yl)- l-methylcarbapen-2-em-3-carboxylate (181 mg, 0.229 mmol) in acetonitrile (4.6 mL) under a nitrogen atmosphere was treated with 1- carbamoylmethyl-4-aza-l-azoniabicyclo[2.2.2]octane triflate (73 mg, 0.229 mmol) followed by a solution of silver trifluoromethanesulfonate in acetonitrile (0.986M, 0.231 mL, 0.228 mmol). The mixture was stirπed in the dark, and after 10 min sonicated in a bath for 10 min and stiπed at room temperature. After 20 min the mixture was filtered through a prewashed celite pad and the filtrate evaporated under vacuum to an oil. The oil was triturated with diethyl ether to give a solid which was pumped under vacuum to give the title compound (262 mg). iH NMR (DMSOdό) δ 0.52(q, J=8.0Hz, OSiCH2C//j), 0.86(t, J=8.0Hz, OSiC//2CH3), 1.03(d, J=7.3Hz, 1-CH3), 1.10(d, J=6.2Hz), 3.5-3.6(m, H-l, H-6), 3.85-3.88(m, DABCO CH2), 4.00-4.05(m, DABCO CH2), 4.22(dq, J=4.1 , 6.2Hz, H-8), 4.25(s, C 2CONH2), 4.33(dd, J=3.2, 10.3Hz, H-5), 4.75(s, ArCH2N), 5.37(d, J=14.2Hz, ArC/ _σ Hb), 5.48(d, J=14.2Hz, ArCH _a b), 7.57(d, J=8.2Hz, ArH), 7.73(d, J=8.7Hz, ArH), 7.8 l(s, CON/ _α Hb), 7.98(s, CONH _a Hb), 8.13(d, J=8.2Hz, ArH), 8.21 (d, J=8.7Hz, ArH), 8.49(s, thiazole H5) IR(KBr) 3422, 2958, 1780, 1706, 1523, 1273 cm-1

Step 5

( 1 R.5S.6S V2f2- ( 3- ( 4-lCarbamoylmethyl- 1.4-diazobicvclor2.2.21 oct-l-vPmethvnphenvπthiazol-2-yl }thiol-6-(l R-hvdroxyethvP-

1 -methylcarbapen-2-em-3-carboxylate chloride

A stiπed solution of 4-nitrobenzyl (l R,5S,6S)-2-[-3 | -4- [carbamoylmethyl- 1 ,4-diazobicyclo[2.2.2]oct- 1 -yl)methyl]phenyl } thiazol-2-yl ) thiol-6( 1 R-triethylsilyloxyeth- 1 -yl)- 1 -methylcarbapen-2-

em-3-carboxylate bis trifluoromethanesulfonate (258 mg, 0.229 mmol) in tetrahydrofuran (4.6 mL) and water (2.3 mL) at room temperature was adjusted using a pH meter to pH 2.3 using 2N hydrochloric acid and maintained at this pH. After 35 min the solution

₅ was adjusted to pH 6.75 using saturated aqueous sodium bicarbonate solution.

The O-desilylated product solution was mixed with n- butanol (4.6 mL), ethyl acetate (2.3 mL), water (4.6 mL), 0.5M pH7 phosphate buffer (2.3 mL), and 10% palladium on carbon (70 mg), _{j 0} nitrogen and hydrogen purged, and vigorously stirred under a balloon of hydrogen at room temperature. After 90 min, the reaction vessel was nitrogen purged and the contents filtered through a prewashed celite pad. The filtrate was allowed to separate into two phases and the aqueous phase was recovered and set aside. The remaining organic phase was

_{: 5} washed with water (2 x 10 mL) and the water washes and original aqueous phase were combined and washed with 1 :1, ethyl acetate :diethyl ether (40 mL). The resulting solution was concentrated under vacuum to approximately half volume and applied to a column (1 10 cm) of Tosohaas Amberchrom® CG-161 resin. The column was water washed

₂₀ (40 mL) and then washed with 40% acetonitrile water to elute the product. The aqueous acetonitrile solution containing product was concentrated under vacuum and treated with ammonium chloride (660 mg, 12.3 mmol) and chromatographed in two portions of a Tosohaas Amberchrom® CG-1000 column (1 x 6 cm) eluting with water. The

₂₅ product in aqueous solution was lyophilized to afford the title compound as an amorphous white fluffy solid (45.3 mg). lH NMR (D20) δ 0.87(d, J=7.4Hz, 1-CH3), 1.13(d, J=6.2Hz, C//JCHOH), 3.17(d, J=7.4, 9.1Hz, H-l,) 3.30(dd, J=2.7, 5.6Hz, H-6), 4.05-4.08(m, DABCO CH2, H-5), 4.1 l(~p, J=~6Hz, H-8), 4.23-4.26(m,

30 DABCO CH2), 4.38(s, CH2CONH2), 4.82(s, ArCH2N), 7.57(d, J=8.3Hz, ArH), 7.84(d, J=8.2Hz, ArH), 7.88(s, thiazole H5) IR(KBr) 3425, 1758, 1701 , 1597 cm-1 UV(0.1M pH7 MOPS buffer) λ _m ax 281nm εl8,400

EXAMPLE 9

(lR.5S.6S -2[2-( 3-(4-r(2-HYDROXYETHYL)-1.4-DIAZOBICYCLO

12.2.21QCT- 1 -YDMETHYL1PHENYL ) THIAZOL-2- YL ) THIOl-6-

( 1 R-HYDROXYETHYL - 1 -METHYLCARB APEN-2-EM-3-

CARBOXYLATE CHLORIDE

Step 1

4-Nitrobenzyl (1 R.5S.6S V2 -3 ( -4-l(2-hvdroxyethvn- 1.4- diazobicvclor2.2.21oct- 1 - vPrnethyllphenyl 1 thiazol-2-yl 1 thiol-6( 1 R-triethylsilylox veth- 1 - vP- 1 -methylcarbapen-2- em-3-carboxylate bistrifluoromethanesulfonate

A stiπed solution of 4 nitrobenzyl-(lR,5S,6S)-2-[4- (4-iodomethylphenyl)thiazol-2-yl)thio]-6-(lR-triethylsilylox yeth- l-yl)-l-methylcarbapen-2-em-3-carboxylate (200 mg, 0.253 mmol) in acetonitrile (3.5 mL) under a nitrogen atmosphere was treated with l-(2-hydroxyethyl)-4-aza-l-azoniabicyclo[2.2.2]octane triflate (77.4 mg, 0.253 mmol) followed by a solution of silver trifluoromethanesulfonate in acetonitrile (0.999M, 0.253 mL, 0.253 mmol). The mixture was stirrred

in the dark. After 35 min the mixture was filtered through a prewashed celite pad and the filtrate evaporated under vacuum to an oil. The oil was triturated with diethyl ether to give a solid which was pumped under vacuum to give the title compound (283 mg). iH NMR (DMSOdό) δ 0.52(q, J=8Hz, OSiCH2CH3), 0.86(t, J=8Hz, OSiCH2CH3), 1.02(d, J=7.4Hz, I-CH3), 1.09(d, J=6.1Hz), 3.5-3.6(m, H- 1, H-6), 3.78-3.95(m, DABCO CH2), 4.22 (dq, J=3.9, 6.1Hz, H-8), 4.33(dd, J=3.2, 10.3Hz, H-5), 4.78(s, ArCH2N), 5.37(d, J=14.1Hz, ArCH _a Hb), 5.48(d, J=14.1Hz, ArCH _a Hb), 7.60(d, J=8.2Hz, ArH), 7.73(d,J=8.5Hz, ArH), 8.12(d,J=8.2Hz,ArH), 8.20(d,J=8.5Hz, ArH), 8.48(s,thiazoleH5) IR(KBr)3450,2959,2877, 1779, 1702, 1523, 1340,1260, 1165 cm-1

Step2 (lR.5S.6SV2r2-(3-f4-r(2-hvdroxyethvP-1.4-diazohicvc or2.2.21 oct- 1 -vPmethvnphenvπthiazol-2-vπ thiol -6-(lR-hydrox vethvP- 1 -methylcarbapen-2-em-3-carboxylate chloride

A stiπed solution of 4-nitrobenzyl (lR,5S,6S)-2-[-3{-4-[(2- hydroxyethyl)-l ,4-diazobicyclo[2.2.2]oct- 1 -yl)methyl]phenyl Jthiazol- 2-y 1 } thio]-6( 1 R-triethylsilyloxyeth- 1 -y 1)- 1 -methylcarbapen-2-em-3- carboxylate bistrifluoromethanesulfonate (278 mg, 0.248 mmol) in tetrahydrofuran (5.0 mL) and water (2.5 mL) at room temperature was adjusted using a pH meter to pH2.3 using 2N hydrochloric acid and maintained at this pH. After 35 min the solution was adjusted to pH6.5 using saturated aqueous sodium bicarbonate solution.

The O-desilylated product solution was mixed with n- butanol (5.0 mL), ethyl acetate (2.5 mL), water (5.0 mL), 0.5M pH7 phosphate buffer (2.5mL), and 10% palladium on carbon (75 mg), nitrogen and hydrogen purged, and vigorously stiπed under a balloon of hydrogen at room temperature. After 90 min, the reaction vessel was nitrogen purged and the contents filtered through a prewashed celite pad. The filtrate was allowed to separate into two phases and the aqueous phase was recovered and set aside. The remaining organic phase was washed with water (2 x 10 mL) and the water washes and original aqueous phase were combined and washed with 1 : 1, ethyl acetate :diethyl

-70-

ether (40 mL). The resulting aqueous solution was concentrated under vacuum and applied to a column (1.5 x 11 cm) of Bio-Rad Macro-Prep CM and eluted with water. After ~6 column volumes of water, the column was washed with 5% aqueous sodium chloride which eluted the

5 product. The saline effluent containing product was applied to a

Tosohaas Amberchrom® CG-161 resin column (1 x 10 cm). The column was water washed (~75 mL) and then washed with 20% isopropanol- water to elute the product. The aqueous isopropanol solution containing product was concentrated under vacuum and lyophilized to afford the title l o compound as an amorphous white fluffy solid (71.2 mg). iH NMR (D20) δ 0.83(d, J=7.3Hz, I-CH3), 1.12(d, J=6.4Hz, CH3CHOH), 3.15(d, J=7.43, 9.4Hz, H-l,) 3.26(dd, J=2.6, 5.6Hz, H-6), 3.72-3.74 (m, CH2OH), 4 .04-4.1 l(m, DABCO CH2, H-5, H-8, NCH2), 4.81(s, ArCH2N), 7.57(d, J=8.1Hz, ArH), 7.83(d, J=8.1Hz, ArH), 7.87(s,

15 thiazole H5)

IR(KBr) 3422, 1758, 1597, 1478, 1390 cm"l UV(0.1M pH7 MOPS buffer) λ _max 281nm εl8,400

EXAMPLE 10 20 (lR.5S.6SV212-(3-(4-r3-METHYLIMIDAZOLIUM-l -

YLMETHYLlPHENYL)THIAZOL-2-YLlTHI01-6-riR- HYDROXYETHYLV 1 -METHYLCARB APEN-2-EM-3-

CARBOXYLATE

25

30

Step l

4-Nitrobenzyl (lR.5S.6SV2-r-3f -4-lcarbamoylmethyl-1.4- diazobicyclor2.2.21oct-l-vPmethvnphenvπthiazol-2-yll thio1-6(l R-triethylsilyloxyeth- 1 -vP- 1 -methylcarbapen- 2-em-3-carboxylate trifluoromethanesulfonate

A stiπed solution of 4 nitrobenzyl-(lR,5S,6S)-2-[4-(4- hydroxymethylphenyl)thiazol-2-yl)thio]-6-( 1 R-triethylsilyloxyeth- l-yl)-l-methylcarbapen-2-em-3-carboxylate (226 mg, 0.331 mmol) in dichloromethane (5.5 mL) under a nitrogen atmosphere was treated with 1-methylimidazole (0.066 mL, 0.829 mmol) and cooled in a dry ice - acetone bath. The cold solution was treated with trifluoromethane¬ sulfonic anhydride (0.070 mL, 0.414 mmol). After 17 min the reaction was removed from the dry ice bath and placed in an ice bath. After 10 min at 0°C the reaction was diluted with dichloromethane (5 mL) and washed with water (2 x 5 mL), dried over magnesium sulfate, filtered and evaporated under vacuum to a foam, which was triturated with benzene and pumped under vacuum to give the title compound (296 mg). iH NMR (CDC13) δ 0.57(q, J=8.0Hz. OSiCH2CH3), 0.91(t, J=8.0Hz, OSiCH2CH3), 1.12(d, J=7.3Hz. I -CH3), 1.20(d, J=6.2Hz), 3.28 (dd,

J=2.9, 4.9Hz, H-6), 3.56 (dq, J=7.3, 9.9Hz, H-l), 3.96 (s, NCH3), 4.26(~p, J=~6Hz, H-8), 4.32 (dd, J=2.9, 9.9Hz, H-5), 5.30(d, J=14Hz, ArCH _a Hb), 5.41(s, ArCH2N), 5.48(d, J=14Hz, ArCHaHb), 7.24 (t, J=2HZ, Im H), 7.27 (t, J=2Hz, Im H), 7.47(d, J=8.2Hz, ArH), 7.64(s, thiazole H5), 7.66(d, J=8.7Hz, ArH), 7.91(d, J=8.2Hz, ArH), 8.20(d, J=8.7Hz, ArH), IR(KBr) 3474, 2957, 2876, 1777, 1708, 1607, 1561, 1522, 1340cm-l

Step 2

(!R.5S.6S )-2[2-f 3-(4-r3-methylimidazolium-l-ylmethyllphenvnthiazol-

2-vπthio1-6-(lR-hvdroxyethyP-l-methylcarbapen-2-em-3-car boxylate

A stiπed solution of 4-nitrobenzyl (lR,5S,6S)-2-[-3{-4- [carbamoylmethyl-1 ,4-diazobicyclo[2.2.2]oct-l -yl)methyl]phenyl } thiazol-2-y 1 } thio]-6( 1 R-triethylsilyloxyeth- 1 -yl)- 1 -methy lcarbapen-2- em-3-carboxylate trifluoromethansulfonate (290 mg, 0.324 mmol) in tetrahydrofuran (4.4 mL) and water (2.2 mL) at room temperature was adjusted using a pH meter to pH2.3 using 2N hydrochloric acid and maintained at this pH. After 35 min the solution was adjusted to pH 6.6 using saturated aqueous sodium bicarbonate solution.

The O-desilylated product solution was mixed with n- butanol (6.4 mL), ethyl acetate (3.2 mL), water (6.4 mL), 0.5M pH7 phosphate buffer (3.2 mL), and 10% palladium on carbon (80 mg), nitrogen and hydrogen purged, and vigorously stiπed under a balloon of hydrogen at room temperature. After 90 min, the reaction vessel was nitrogen purged and the contents filtered through a prewashed celite pad. The filtrate was allowed to separate into two phases and the aqueous phase was recovered and set aside. The remaining organic phase was washed with water (2 x 10 mL) and the water washes and original aqueous phase were combined and washed with 1 :1, ethyl acetate :diethyl ether (40 mL). The resulting solution was concentrated under vacuum to approximately half volume and chromatographed on a Tosohaas Amberchrom® CG-lOOOsd column (1.5 x 1 1 cm) eluting first with water followed by a gradient elution of acetonitrile - water. The product eluted in 12-14% acetonitrile - water. The product fraction was concentrated

and the solution was lyophilized to afford the title compound as an amorphous white fluffy solid (75.4 mg). lH NMR (D2θ) δ 0.64(d, J=7.1Hz, I-CH3), 1.02(d, J=6.4Hz,

CH3CHOH), 2.0(dq, J=7.4, 9.1Hz, H-l,) 3.10(dd, J=2.5, 5.5Hz, H-6), 3.82 (s, NCH3), 3.89 (dd, J=2.5, 9.6Hz, H-5), 4.00(~p, J=~6Hz, H-8),

4.38(s, CH2CONH2), 5.32(s, ArCH2N), 7.32(d, J=8.1Hz, ArH), 7.38 (s,

Im H), 7.43 (s, Im H), 7.52(s, thiazole H5), 7.56(d, J=8.1Hz, ArH), 8.86

(s, Im H-2)

IR(KBr) 3416, 1756, 1599, 1386, 1280, 1160 cm-1 UV(0.1 M pH7 MOPS buffer) λmax 266nm εl7,500

EXAMPLE 11 (1R.5S.6S 2Γ2-( 3-( 3-ΓCARBAMOYLMETHYL-1.4- DIAZOBICYCLOΓ2.2.21QCT-1 -YL)METHYL1PHENYL)THIAZOL-2- YL 1 THIQ1-6-( 1 R-HYDROX YETHYL 1 -METHYLC ARB APEN-2-

EM-3-CARBOXYLATE CHLORIDE

R = OH ■

___ I MsCI. Et-jN /CH2CI2

R = OMs "*~~^ l / acetone

R = l ι Na

Step 1

4-Nitrobenzyl (lR.5S.6SV2-[4-(4-hvdroxymethylphenvP thiazol-2-yPthiol-6-(lR-triethylsilyloxyeth-l -vP-l- methylcarbapen-2-em-3-carboxylate

5 A suspension of powdered lithium hydroxide monohydrate

(36 mg, 0.86 mmol) in dry tetrahydrofuran (7.2 mL) at room temperature was treated with 4-(3-hydroxymethylphenyl)-2-mercaptothiazole (160 mg, 0.717 mmol) to give a light yellow solution. Solid 4-nitrobenzyl (1R,5R, 6S)-2-trifluoromethanesulfonyloxy)-6-(lR-triethylsilyloxyeth - l o l -yl). l -methylcarbapen-2-em-3-carboxylate (445 mg, 0.731 mmol) was added to the solution. After stirring 65 min the reaction was diluted with ethyl acetate (15 mL) and washed with water (15 mL), brine, dried over magnesium sulfate, filtered, and evaporated under vacuum to a yellow oil (647 mg). The crude product was flash silica gel column (2 x 16 cm)

15 chromatographed eluting with 1 :1, ethyl acetate.hexane. Product containing fractions were combined, evaporated under vacuum, and lyophilized from benzene to give the title compound as an amorphous white solid (412 mg). lH NMR (CDCl3) δ 0.58 (q, J=8.0Hz, S1CH2CH3)), 0.92(t, J=8.0Hz,

20 S1CH2CH3), 1.12(d, J=7.4Hz, I-CH3), 1.21(d, J=6.1Hz, C 3CHOH), 3.27(dd, J=3.0, 5.3Hz, H-6), 3.60(dq, J=7.4, 9.9Hz, H-l), 4.26(~p, J=6Hz, CH3C//OH), 4.31(dd, J=3.0, 9.9Hz, H-5), 5.3 l(d, J=13.7Hz, C02C// _α Hb), 5.50(d, J=13.7Hz, Cθ2CH _a //fc), 7.38(d, J=7.8Hz, H-4), 7.44(d, J=7.7Hz, H-5), 7.60 (s, thiazole H-5), 7.67(d, J=8.7Hz, ArH2),

25 7.81(d, J=7.7Hz, H-2), 8.22(d, J=8.7Hz, ArH2)

IR(KBr) 3449,2956, 1777, 1708, 1607, 1552, 1340cm-1 MS(CI)(m/z)682.0(M+l)

Step 2 ₃₀ 4-Nitrobenzyl (lR.5S.6SV2-f(4-(3-methanesulfonyloxy- methylphenyPthiazol-2yPthiol-6-( 1 R-triethylsilylox veth- 1 -yP- 1 -methylcarbapen-2-em-3-carboxylate

A solution of 4-nitrobenzyl (lR,5S,6S)-2[(4-(3-hydroxy- methylpheny l)thiazol-2-yl)thio]-6-( 1 R-triethylsilyloxyeth- 1 -yl)- 1 - methylcarbapen-2-em-3-carboxylate (405 mg, 0.594 mmol) in dry

dichloromethane (6 mL) under a nitrogen atmosphere was cooled in an ice bath. The cold solution was treated with triethylamine (0.124 mL, 0.891 mmol) follwed by methanesulfonyl chloride (0.057 mL, 0.742 mmol). After 30 min the solution was diluted with dichloromethane (4 mL) and washed with water (5 mL), 0.5M HCl (5 mL), and water (2 x 5 mL), dried over magnesium sulfate, filtered, and evaporated to afford the title compound as an oil (421 mg). lH NMR (CDCl3) δ 0.60 (q, J=8.0Hz, S1CH2CH3)), 0.92(t, J=8.0Hz, SiC 2CH3), 1.13 (J=7.3Hz,l-CH3), 1.22(d, J=6.1Hz, C//5CHOH), 0 2.98(s, CH3SO2), 3.28(dd, J=3.0., 5.3Hz, NCH), 3.60 (dq, J=7.3,10.0Hz, H-l), 4.27(~p, J=~6Hz, CH3C /OH), 4.33(dd, J=3.0, 10.0Hz, H-5), 5.30(s, Ar-CH2), 5.31(d, J=13.9Hz, Cθ2C _α Hb), 5.50(d, J=13.9Hz, C02CH _a //£), 7.43(d, J=7.8Hz, ArH), 7.49 (t, J=7.8Hz, H-5), 7.63(s, thiazole H5), 7.67(d, J=8.7Hz, ArH2), 7.92(d, J=8.3Hz, ArH), 7.94 (d, 5 J=7.8Hz, H-2), 8.22 (d, J=8.7Hz, ArH2)

IR(KBr)3448,2956,2876, 1778, 1708, 1523, 1340cm-1 MS(CI) /z 760.1 (M+l)

Step3 ₀ 4-Nitrobenzyl (lR5S.6SV2-r4-(3-iodomethylphenvPthiazol-2-vPthiol-6- ( 1 R-triethylsil yloxyeth- 1 -vP- 1 -methylcarbapen-2-em-3-carboxylate A stiπed solution of 4-nitrobenzyl (lR,5S,6S)-2-[(4-(3- methanesulfonyloxymethylphenyl)thiazol-2-yl)thio]-6-(lR-trie thylsilyl- oxyeth-l-yl)-l-methylcarbapen-2-em-3-carboxylate (418 mg, 0.550

₂₅ mmol) in acetone (5.5 mL) was treated with sodium iodide (247 mg, 1.65 mmol) and stiπed at room temperature. After 70 min the reaction mixture was partitioned between ethyl acetate (15 mL) and water (15 mL). The organic phasw was reovered and washed with water (15 mL), brine, dried over magnesium sulfate, filtered, and evaporated under

_ ₀ vacuum to a foam (432 mg). The crude product was flash silica gel column (2 x 13 cm) chromatographed eluting with 5:2, hexane:ethyl acetate. Product containing fractions were combined and evaporated under vacuum to a partial solid and lyophilized from benzene to afford the title compound (408 mg) as a white solid.

lH NMR (CDCl3) δ 0.58 (q, J=8.0Hz, S1CH2CH3), 0.92(t, J=8.0Hz, SiC/ 2CH3), 1.12(d, J=7.3Hz, I-CH3), 1.22(d, J=6.2Hz, C/ 5CHOH), 3.27(dd, J=2.8 5.4Hz, H-6), 3.61(dq, J=7.3, 9.9Hz, H-l ) , 4.26(~p, J=~6Hz, CH3C//OH), 4.32(dd, J=2.8, 9.9Hz, H-5), 4.51(s, ArCH2), 5.31(d, J=13.7Hz, Cθ2C// _α Hb), 5.50(d, J=13.7Hz, Cθ2CH _a /b), 7.36- 7.42(m, ArH), 7.60(s, thiazole H5), 7.67(d, J=8.7Hz, ArH2), 7.75(td, J=2, 7Hz, ArH), 8.23(d, J=8.7Hz, ArH2) IR(KBr) 3449, 2956, 2875, 1779,1708, 1522, 1340 cm-1 MS m/z 792.1 (M+l)

Step 4

4-Nitrobenzyl (lR.5S.6S)-2-l-4( -3-fcarbamoylmethyl-l .4- diazobicvclo[2.2.21oct- 1 -vPmethyllphenyl lthiazol-2-yl ) thiol-6( 1 R-triethylsil yloxyeth- 1 -vP- 1 -methylcarbapen- 2-em-3-carboxylate bistrifluoromethanesulfonate

A stiπed solution of 4 nitrobenzyl-(lR,5S,6S)-2-[4-(3- iodomethylphenyl)thiazol-2-yl)thio] -6-( 1 R-triethylsilyloxyeth- 1 - yl)-l-methylcarbapen-2-em-3-carboxylate (220 mg, 0.253 mmol) in acetonitrile (5.3 mL) under a nitrogen atmosphere was treated with 1- carbamoylmethyl-4-aza-l -azoniabicyclo[2.2.2]octane trifluorosulfonate (84 mg, 0.263 mmol) and sonicated (5 min) to give a solution. The solution was treated with a solution of silver trifluoromethanesulfonate in acetonitrile (0.999 M, 0.263 mL, 0.263 mmol) and the mixture stirπed in the dark. After 30 min the mixture was sonicated (5 min) and filtered through a prewashed celite pad and the filtrate evaporated under vacuum to give the title compound (296 mg) as a white solid.. iH NMR (DMSOdό) δ 0.52(q, J=8.0Hz, OSiCH2C//j), 0.86(t, J=8.0Hz, OSiC//2CH3), 1.03(d, J=7.1Hz, I-CH3), 1.09(d, J=6.2Hz), 3.45-3.54(m, H-l , H-6), 3.82-3.92(m, DABCO CH2), 9.98-4.06(m, DABCO CH2), 4.18-4.26(m H-8, H-5)), 4.24(s, CH2CONH2), 4.79(s, ArCH2N), 5.37(d, J=14Hz, ArC/ _fl Hb), 5.48(d, J=14Hz, ArCH _a Hb), 7.50(d, J=7.8Hz, ArH), 7.65(d, J=7.8Hz, ArH), 7.73(d, J=8.5Hz, ArH2), 7.81(s, CONH _a Rb), 7.97(s, CONH _a b), 8.10(s, ArH), 8.15(d, J=7.8Hz, ArH), 8.21 (d, J=8.5Hz, ArH2), 8.49(s, thiazole H5) IR(KBr) 3432, 2959, 2877, 1779, 1702, 1523, 1341 , 1275 cm-1

Step 5

(1 R.5S.6S V2f2- ( 3- ( 3-lCarbamoylmethyl- 1 ,4-diazobicvclo r2.2.21oct-l-vPmethvnphenyllthiazol-2-yl)thiol-6-(lR- hvdroxyethvP- 1 -methylcarbapen-2-em-3-carboxylate chloride A stiπed solution of 4-nitrobenzyl (1 R,5S,6S)-2-[-3 { -3

[carbamoylmethyl-1 ,4-diazobicyclo[2.2.2]oct- 1 -yl)methyl]phenyl } thiazol-2-yl } thio]-6(l R-triethylsilyloxyeth- 1 -yl)- 1 -methylcarbapen-2- em-3 -carboxylate bistrifluoromethanesulfonate (287 mg, 0.253 mmol) in tetrahydrofuran (5.0 mL) and water (2.5 mL) at room temperature was adjusted using a pH meter to pH2.3 using 2N hydrochloric acid and maintained at this pH. After 35 min the solution was adjusted to pH6.5 using saturated aqueous sodium bicarbonate solution.

The O-desilylated product solution was mixed with n-butanol (5.0 mL), ethyl acetate (2.5 mL), water (5.0 mL), 0.5M pH7 phosphate buffer (2.5 mL), and 10% palladium on carbon (80 mg), nitrogen and hydrogen purged, and vigorously stiπed under a balloon of hydrogen at room temperature. After 90 min, the reaction vessel was nitrogen purged and the contents filtered through a prewashed celite pad. The filtrate was allowed to separate into two phases and the aqueous phase was recovered and set aside. The remaining organic phase was washed with water (10 mL) and the water wash and original aqueous phase were combined and washed with 1 :1, ethyl acetate:diethyl ether (25 mL). The washed aqueous solution was concentrated under vacuum and applied to a column (2 x 5 cm) of Bio-Rad Macro-Prep CM resin. The column was water washed (ca. 50 mL) and then washed with 5% aqueous sodium chloride to elute the product. Product containing fractions were combined and concentrated under vacuum (ca. 10 mL) and chromatographed on a Tosohaas Amberchrom® CG-1000 column (1.5 x 11 cm) eluting first with water followed by a gradient elution of acetonitrile - water. The product eluted with 6% acetonitrile in water. The aqueous acetonitrile solution containing product was concentrated under vacuum and lyophilized to afford the title compound as an amorphous white fluffy solid (7 mg).

lH NMR (D2θ) δ 0.87(d, J=7.4Hz, I-CH3), 1.13(d, J=6.2Hz, O/jCHOH), 3.17(d, J=7.4, 9.1Hz, H-l,) 3.30(dd, J=2.7, 5.6Hz, H-6), 4.05-4.08(m, DABCO CH2, H-5), 4.1 l(~p, J=~6Hz, H-8), 4.23-4.26(m, DABCO CH2), 4.38(s, CH2CONH2), 4.82(s, ArCH2N), 7.57(d, J=8.3Hz, ArH), 7.84(d, J=8.2Hz, ArH), 7.88(s, thiazole H5) IR(KBr) 3425, 1758, 1701, 1597 cm-1 UV(0.1M pH7 MOPS buffer) λ _ma χ 317nm ε9,700

EXAMPLE 12 ( 1 R.5S.6SV 2f2- f 3- ( 3-r3-METHYLIMIDAZOLIUM- 1 - YDMETHYL1 PHENYL ) THIAZOL-2- YL I THIQ1-6-( 1 R-HYDROX YETHYLV 1 - METHYLCARBAPEN-2-EM-3-CARBOXYLATE

Step l

4-Nitrobenzyl ( 1 R.5S.6S)-2-f-4 ( -3-13-methylimidazolium- 1 -vP methyllphenvπ thiazol-2-yl 1 thiol -6( 1 R-triethylsilyloxyeth- 1 -vP- 1 -methylcarbapen-2-em-3-carboxylate trifluoromethanesulfonate

A stiπed solution of 4 nitrobenzyl-(lR,5S,6S)-2-[4-(3- iodomethylphenyl)thiazol-2-yl)thio]-6-( 1 R-triethylsilyloxyeth- 1 -yl)- l-methylcarbapen-2-em-3-carboxylate (192 mg, 0.242 mmol) in acetonitrile (4.8 mL) under a nitrogen atmosphere was treated with 1-methylimidazole (0.020 mL, 0.254 mmol) followed by a solution

of silver trifluoromethanesulfonate in acetonitrile (0.999 M, 0.241 mL, 0.241 mmol) and the mixture stirrred in the dark. After 30 min the mixture was sonicated (5 min) and filtered through a prewashed celite pad and the filtrate concentrated under vacuum and lyophilized from benzene to give the title compound (269 mg) as a solid. iH NMR (DMSOdό) δ 0.51(q, OS1CH2CH3), 0.86(t, OSiO/2CH3), 1.03(d, I-CH3), 1.09(d, C/ 5CHOH), 4.79(s, A1CH2N), 5.37(d, ArC/ _α Hb), 5.48(d, ArCH _a b), 7.73(d, ArH2), 8.10(s, ArH), 8.21 (d, ArH2), 9.20 (s, Im H-2)

Step 2

( 1 R.5S.6S V2I - (3-13-r3-methylimidazolium- 1 -vPmethyllphenyl ) thiazol-2-vπthio1-6-(lR-hvdroxyethvP-l-methylcarbapen-2-em- 3- carboxylate

A stiπed solution of 4-nitrobenzyl (lR,5S,6S)-2-[-3{- 3[3-methylimidazolium- 1 -yl)methyl]phenyl } thiazol-2-yl } thio]-6( 1 R- triethylsilyloxyeth- 1 -yl)- 1 -methylcarbapen-2-em-3-carboxylate trifluoro¬ methanesulfonate (216 mg, 0.242 mmol) in tetrahydrofuran (5.0 mL) and water (2.5 mL) at room temperature was adjusted using a pH meter to pH 2.3 using 2N hydrochloric acid and maintained at this pH. After 35 min the solution was adjusted to pH6.5 using saturated aqueous sodium bicarbonate solution.

The O-desilylated product solution was mixed with n- butanol (5.0 mL), ethyl acetate (2.5 mL), water (5.0 mL), 0.5M pH7 phosphate buffer (2.5 mL), and 10% palladium on carbon (55 mg), nitrogen and hydrogen purged, and vigorously stiπed under a balloon of hydrogen at room temperature. After 90 min, the reaction vessel was nitrogen purged and the contents filtered through a prewashed celite pad. The filtrate was allowed to separate into two phases and the aqueous phase was recovered and set aside. The remaining organic phase was washed with water (15 mL) and the water wash and original aqueous phase were combined and washed with 1 :1 , ethyl acetate:diethyl ether (40 mL). The resulting solution was concentrated under vacuum and chromatographed on a Tosohaas Amberchrom® CG-lOOOsd column (1.5 x 1 1 cm) eluting first with water. The column was gradiently eluted with

acetonitrile water and the product eluted in ca. 16% acetonitrile/water. The aqueous acetonitrile solution containing product was concentrated under vacuum and lyophilized to afford the title compound as an amorphous white fluffy solid (7.8 mg). !H NMR (D20) δ 1.01(d, J=7.3Hz, I-CH3), 1.19(d, J=6.4Hz,

C 3CHOH), 3.20(dq, J=7.3 9.7Hz, H-l,) 3.40(dd, J=2.9, 6.1Hz, H-6), 3.83 (s, NCH3), 4.15 (dd, J=2.6, 6.9Hz, H-5), 4.16(~p, J=~6Hz, H-8), 5.41(s, ArCH2N), 7.40 (d, J=8Hz, Ar H-6), 7.41 (t, J=2Hz, Im H), 7.46 (t, J=2Hz, Im-H), 7.51 (t, J=8Hz, Ar H-5), 7.77 (s, Ar H-2), 7.81 (d, J=8Hz, Ar H-4), 7.85(s, thiazole H5), 8.76 (s, Im H-2) IR(KBr) 3448, 1752, 1608, 1560, 1397 cm-1 UV(0.1M pH7 MOPS buffer) λ _max 312nm ε8,180

EXAMPLE 13 (lR.5S.6S 2-f(4-PHENYL-(5-fCARBAMOYLMETHYL-1.4-

DIAZABICYCLθr2.2.210CT-l-YL)METHYL^THIAZOLO-2-THI01

-6-r 1 (R HYDROXYETHYL1- 1 -METHYLCARB APEN-2-EM 3-

CARBOXYLATE CHLORIDE

Step l p-Nitrobenzyl (lR.5S.6SV2-[4-phenyl-(5-(hvdroxymethvP thiazolo-2-thio1-6- ( 1 R triethylsil ylox veth- 1 -yll - 1 - methylcarbapen-2-em-3-carboxylate

A mixture of 4-phenyl-(5-(hydroxymethyl)-2- mercaptothiazole (lOOmg, 0.448mmol), p-Nitrobenzyl (1R,5R,6S)- 2-(trifluoromethanesulfonyloxy)-6-[( 1 R)-triethylsilyloxyeth- 1 -yl]- l-methylcarbapen-2-em-3-carboxylate (272 mg, 0.448 mmol) and powdered lithium hydroxide monohydrate (24 mg, 0.57 mmol) in tetrahydrofuran (3 mL) was stiπed at room temperature for 75 minutes under nitrogen. The suspension was partitioned between ethyl acetate

(30 mL) and 5% aqueous sodium bicarbonate (20 mL). The ethyl acetate layer was washed with brine (20 mL), dried with magnesium sulfate, filtered and evaporated to give an oil (150 mg), which contained residual tetrahydrofuran as observed by NMR. The oil was purified on preparative TLC plates (2x1000 micron, eluted with 10% ethyl acetate/ methylene chloride) to give the title compound as a foam (220mg). -H NMR (CDC1 ₃ , 400 MHz) δ 0.61 (q, Si(CH2£H ₃ ) ₃ , 0.93 (t, Si(£H2CH ₃ ) ₃ , 1.12 (d, 1-CH ₃ ), 1.22 (d, CH3CHOH), 3.26 (dd, H-6), 3.68 (dq, H-l), 4.26 (p, CH ₃ CHOH), 4.34 (dd, H-5), 4.90 (ABq, CH ₂ OH), 5.31 and 5.50 (two d's, CH ₂ C ₆ H4NO ₂ ), 7.45 (m, 5ArH), 7.66 and 8.20

Step 2 p-Nitrobenzyl (lR.5S.6SV2-r4-phenyl-(5-(iodomethvPthiazolo-2-thiol-6- \( lRVtriethylsilyloxyeth-1 -yll- 1 -methylcarbapen-2-em-3-carboxylate A solution of p-Nitrobenzyl (lR,5S,6S)-2-[4-phenyl- (5-(hydroxymethyl)thiazolo-2-thio]-6-[(l R)-triethylsilyloxyeth- 1 -yl]- l-methylcarbapen-2-em-3-carboxylate (220 mg, 0.323 mmol) and triethylamine (0.063 mL, 0.45mmol) in methylene chloride (6 mL) was cooled in an ice bath under a nitrogen atmosphere. Methanesulfonyl chloride (0.031 mL, 0.404 mmol) was added dropwise over 1 minute. After 1 hour, the mixture was partitioned between methylene chloride (6 mL) and 0.1N pH7 phosphate buffer (12 mL). The aqueous layer was re-extracted with methylene chloride (2 x 10 mL) and the combined methylene chloride layers were dried with magnesium sulfate, filtered and evaporated to give the crude mesylate as an oil (216 mg). The oil was dissolved in acetone (4 mL) and sodium iodide (100 mg, 0.654 mmol) was added. After 55 minutes, the mixture was partitioned between ethyl acetate (30 mL) and water (20 mL). The ethyl acetate layer was washed with 5% aqueous sodium bisulfite (20 mL) and brine (20 mL) and was then dried with magnesium sulfate, filtered and evaporated to give the title compound as an oil (240 mg). ⁱ H NMR (CDCI ₃ , 500 MHz) δ 0.60 (q, Si(CH2CH3) ₃ , 0.96 (t, Si(CH ₂ CH ₃ ) ₃ , 1.16 (d, I -CH ₃ ), 1.23 (d, CH3CHOH), 3.25 (dd, H-6), 3.70

(dq, H-l),4.25 (p, CH ₃ CHOH),4.35 (dd,H-5),4.75 (s,CH ₂ I),5.30and 5.50(twod's,CH ₂ C ₆ H ₄ NO ₂ ),7.50(m,5ArH),7.67and8.22(twom's, CH2C6H4NO2).

p-Nitrobenzyl (lR.5S.6SV2-r(4-phenyl-5-(carbamoylmethyl-l .4- diazabicvclor2.2.21oct-l -vPmethvPthiazolo-2-thiol-6-[ 1 (RV triethylsilyloxyeth-l-yll-l-methylcarbapen-2-em 3-carboxylate bis trifluoromethanesulfonate A solution of p-Nitrobenzyl ( lR,5S,6S)-2-[4-phenyl-5-(iodomethyl) thiazolo-2-thio]-6-[(lR)-triethylsilyloxyeth-l -yl]-l -methylcarbapen- 2-em-3-carboxylate (210 mg, 0.265 mmol) and carbamoylmethyl- 1 ,4-diazabicyclo[2.2.2]octane ditrifluromethanesulfonate (85 mg, 0.265mmol) in acetonitrile (3 mL) was treated with an acetonitrile solution of silver trifluoromethane sulfonate (0.265 mL, 0.999 M in acetonitrile). After 30 minutes at room temperature, the suspension was filtered through pre-washed solka floe, the filtrate was evaporated and the residue was triturated with diethyl ether (10 mL). The resulting precipitate was filtered and provided the title compound as an amorphous solid (280 mg).

Step 4

( 1 R.5S.6S V2-r(4-phenyl-5-(carbamoylmethyl- 1.4-diazabicvclor2.2.21oct- 1 -vPmethvPthiazolo-2-thiol-6-r 1 (R)-hvdroxyethyll- 1 -methylcarbapen-2- em 3-carboxylate chloride

The pH of a solution of p-Nitrobenzyl (lR,5S,6S)-2-[(4- pheny l-5-(carbamoy lmethyl- 1 ,4-diazabicyclo[2.2.2]oct- 1 -yl)methy 1) thiazolo-2-thio]-6-[ 1 (R)-triethylsilyloxyeth- 1 -yl] - 1 -methylcarbapen-2- em 3-carboxylate bis trifluoromethanesulfonate (280 mg, 0.26 mmol) in a mixture of tetrahydrofuran (6 mL) and water (3 mL), was adjusted to 2.40 with IN trifluoromethanesulfonic acid. After stirring for 35 minutes at room temperature, the pH was adjusted to 5.5 with 5% aqueous sodium bicarbonate. Butanol (7 mL), ethyl acetate (5 mL), and 0.1N pH7 sodium phosphate buffer ( 16 mL) were added and the rapidly stiπed mixture was

hydrogenated (atmospheric pressure) in the presence of 10% palladium on carbon (90mg). After 2 hours, the mixture was filtered through a 0.45 micron acrodisc, the aqueous layer was removed and the organic layer was extracted with water (1 x 5 mL). The combined aqueous layers were sparged with nitrogen and the aqueous layer was loaded onto a weak cation exchange column (1x20 cm of Bio-rad macroprep cm support). The column was eluted with water (6x15 mL) and then with 5% aqueous sodium chloride. Fractions (15 mL each) were collected and the product eluted in fractions 1-4 of the 5% aqueous sodium chloride eluent. The combined fractions were loaded onto an amberchrom column (8 mL), and the column was washed with water (75 mL) and then eluted with 20% isopropanol/ water (30 mL). The isopropanol/ water eluent was evaporated to ca. ImL and was lyophilized to give the title compound as a white solid (18 mg). UV (water) λ _m ∞ 285 nm (ε 12,040).

-H NMR (D ₂ 0, 500 MHz) δ 1.15 (d, 1-CH ₃ ), 1.24 (d, CH ₃ CHOH), 3.46 (dq, H-l), 3.56 (dd, H-6), 3.80 and 4.10 (m, CH ₂ N), 4.22 (p, CH ₃ CHOH), 4.25 (dd, H-5), 5.13 (s, CH ₂ Ar) and 7.56 (m, 5 ArH).

EXAMPLE 14

(lR.5S.6SV2-r(4-((5-CARBAMOYLMETHYL-1.4-

DIAZABICYCLOΓ2.2.21QCT- 1 -YDMETHYL^THIEN-2-

YL)THIAZOLO-2-THI01-6-r 1 (R VHYDROX YETHYL1- 1 -

METHYLCARBAPEN-2-EM 3-CARBOXYLATE CHLORIDE

Step l p-Nitrobenzyl ( 1 R.5S.6S V 2-r4-(5-(hvdrox vmethvPthien-2-vPthiazolo-

2-thio]-6-[( 1 RVtriethylsilyloxyeth- 1 -yll- 1 -methylcarbapen-2-em-3- carboxylate

A mixture of 4-(5-(hydroxymethyl)thien-2-yl)-2- mercaptothiazole (70 mg, 0.305 mmol), p-Nitrobenzyl (1R,5R,6S)- 2-(trifluoromethanesulfono)oxy-6-[( 1 R)-triethylsilyloxyeth-l -yl]- l-methylcarbapen-2-em-3-carboxylate (186 mg, 0.305 mmol) and powdered lithium hydroxide monohydrate (15 mg, 0.366mmol) in tetrahydrofuran was stiπed at room temperature for 2 hours under nitrogen. The suspension was partitioned between methylene chloride

(10 mL) and 5% aqueous sodium bicarbonate (20 mL). The aqueous layer was re-extracted with more methylene chloride (2x lOmL), and the combined methylene chloride extracts were dried with magnesium sulfate, were filtered and evaporated to give the title compound as an oil (150 mg), which contained residual tetrahydrofuran as observed by NMR. ⁱ H NMR (CDC1 ₃ , 400 MHz) δ 0.60 (q, Si(CH2£H ₃ ) ₃ , 0.93 (t, Si(£H ₂ CH ₃ ) ₃ , 1.12 (d, I-CH ₃ ), 1.23 (d, CH ₃ CHOH), 3.28 (dd, H-6), 3.68 (dq, H-l), 4.27 (p, CH ₃ CHOH), 4.34 (dd, H-5), 4.83 (ABq, CH ₂ OH), 5.30 and 5.49 (two d's, CH2C ₆ H4NO2), 6.96 and 7.32 (d, 2ArH), 7.39 (s, ArH), 7.67 and 8.22 (two d,

Step 2 p-Nitrobenzyl (lR.5S.6SV2-r4-(5-(iodomethvPthien-2-yPthiazolo-2- thio1-6-[(lR)-triethylsilyloxyeth-l -yll-1 -methylcarbapen-2-em-3- carboxylate

A solution of p-Nitrobenzyl (lR,5S,6S)-2-[4-(5-(hydroxy- methyl)thien-2-yl)thiazolo-2-thio]-6-[(l R)-triethylsilyloxyeth- 1 -yl] - 1 - methylcarbapen-2-em-3-carboxylate (150 mg, 0.218mmol) and triethyl- amine (0.043 mL, 0.305 mmol) in methylene chloride (3 mL) was cooled in an ice bath under a nitrogen atmosphere. Methanesulfonyl chloride (0.021 mL, 0.273 mmol) was added dropwise over 1 minute. After 25 minutes, the mixture was partitioned between methylene chloride (6 mL) and 0.1N pH7 phosphate buffer (6 mL). The aqueous layer was re-extracted with methylene chloride (2 x 10 mL) and the combined methylene chloride layers were dried with magnesium sulfate, filtered and evaporated to give the crude mesylate as an oil (146 mg). The oil was dissolved in acetone (4 mL) and sodium iodide (100 mg, 0.654 mmol) was added. After 40 minutes, the mixture was partitioned between ethyl acetate (30mL) and water (20 mL). The ethyl acetate layer was washed with 5% aqueous sodium bisulfite (20 mL), brine (15 mL) and was dried with magnesium sulfate, filtered and evaporated to give the title compound as an oil (150 mg).

Η NMR (CDCI ₃ , 500 MHz) δ θ.61 (q, Si CH Qh , 0.94 (t, Si(CH ₂ CH ₃ ) ₃ , 1.12 (d, I-CH ₃ ), 1.24 (d, CH ₃ CHOH), 3.29 (dd, H-6), 3.69 (dq, H-l), 4.27 (p, CH ₃ CHOH), 4.32 (dd, H-5), 4.74 (s, CH ₂ I), 5.30 and 5.50 (two d's, CH ₂ C ₆ H ₄ NO ₂ ), 7.10 and 7.24 (d, CH ₂ Ar), 7.42 (s, ArH), 7.67 and 8.22 (two m's, CH ₂ C ₆ H4NO2).

Step3 p-Nitrobenzyl (lR.5S.6SV2-l(4-((5-carbamoylmethyl-l .4- diazabicvclor2.2.21oct-l-vPmethvPthien-2-vPthiazolo-2- thio1-6-ri(RVtriethylsilyloxyeth-l-yll-l-methylcarbapen- 2-em 3-carboxylate bis trifluoromethanesulfonate

A solution of p-Nitrobenzyl (lR,5S,6S)-2-[4-(5- (iodomethyl)thien-2-yl)thiazolo-2-thio]-6-[(lR)-triethylsily loxyeth- l-yl]-l-methylcarbapen-2-em-3-carboxylate (155 mg, 0.19 mmol) and carbamoylmethyl- 1 ,4-diazoniabicyclo[2.2.2]octane ditrifluromethane- sulfonate (60.8 mg, 0.19 mmol) in acetonitrile (2 mL) was treated with an acetonitrile solution of silver trifluoromethane sulfonate (0.19 mL, 0.999M in acetonitrile). After 1 hour at room temperature, the suspension was filtered through pre washed solka floe, the filtrate was evaporated and the residue was triturated with diethyl ether (10 mL). The resulting precipitate was filtered and provided the title compound as an amorphous solid (190 mg).

Step 4 (lR.5S.6SV2-r(4-((5-carbamoylmethyl-1.4-diazabicvclor2.2.21o ct-l - vPmethvPthien-2-vPthiazolo-2-thiol-6-r 1 (R)-hvdroxyethyll- 1 - methylcarbapen-2-em 3 -carboxylate chloride

The pH of a solution of p-Nitrobenzyl (lR,5S,6S)-2-[(4-((5- carbamoylmethyl- 1 ,4-diazoniabicyclo[2.2.2]oct- 1 -yl)methyl)thiophen-2- yl)thiazolo-2-thio]-6-[l(R)-triethylsilyloxyeth-l-yl]-l-meth ylcarbapen-2- em 3-carboxylate bis trifluoromethanesulfonate (190 mg, 0.167 mmol) in a mixture of tetrahydrofuran (3 mL) and water (2 mL), was adjusted to 2.40 with IN trifluoromethanesulfonic acid. After stirring for 35 minutes at room temperature, the pH was adjusted to 6.0 with 5% aqueous sodium bicarbonate. Butanol (4 mL), ethyl acetate (2 mL), and 0.1N pH7 sodium

phosphate buffer (9 mL) were added and the rapidly stirred mixture was hydrogenated (atmospheric pressure) in the presence of 10% palladium on carbon (50 mg). After 2 hours, the mixture was filtered through a 0.45 micron acrodisc, the aqueous layer was removed and the organic layer was extracted with water (1x5 mL). The combined aqueous layers were sparged with nitrogen and the aqueous layer was loaded onto a weak cation exchange column (18mL of Bio-rad macroprep cm support). The column was eluted with water (5x6mL) and then with 1 % aqueous sodium chloride. Fractions (6 mL each) were collected and the product eluted in fractions 3-10 of the 1 % aqueous sodium chloride eluent. The combined fractions were loaded onto an amberchrom column (6 mL), and the column was washed with water (5x6 mL) and then eluted with 25% methanol/ water (18 mL). The methanol/ water eluent was evaporated to ca. 1 mL and was lyophilized to give the title compound as a white solid (30 mg).

UV (water) λ _m a _x 302 nm (ε 21,600). iH NMR (D ₂ 0, 500 MHz) δ 0.99 (d, 1-CH ₃ ), 1.18 (d, CHjCHOH), 3.22 (dq, H-l), 3.39 (dd, H-6), 4.09 and 4.26 (m, CH ₂ N), 4.09 (p, CH ₃ CHOH), 4.16 (dd, H-5), 4.39 (s, CH ₂ Ar), 5.04 (s, CH ₂ C(0)NH2), 7.40 and 7.47 (d, ArH) and 7.83 (s, ArH).

EXAMPLE 15 (lR.5S.6SV2-r(4-((5-CARBAMOYLMETHYL-1.4-

DIAZABICYCLOΓ2.2.21OCT- 1 -YDETHYLYΓHIEN-2- YL^THI AZOLO-2-THIQ1-6-Γ 1 (R VHYDROX YETHYL1- 1 -

METHYLCARBAPEN-2-EM 3-CARBOXYLATE CHLORIDE

4) weakly acidic cation exchange

Sttp i p-Nitrobenzyl (lR.5S.6S)-2-r4-(5-fhvdroxyethvPthien-2-vPthiazolo- 2-thiol-6-f(lRVtriethylsilyloxyeth-l-yll-l -methylcarbapen-2-em-3- carboxylate

A mixture of 4-(5-(hydroxyethyl)thien-2-yl)-2- mercaptothiazole (110 mg, 0.448 mmol), p-Nitrobenzyl (1R,5R,6S)- 2-(trifluoromethanesulfono)oxy-6-[( 1 R)-triethylsiIy loxyeth- 1 -yl]- 1 - methylcarbapen-2-em-3-carboxylate (272 mg, 0.448 mmol) and powdered lithium hydroxide monohydrate (24 mg, 0.57 mmol) in tetrahydrofuran (3 mL) was stiπed at room temperature for 1 hour under nitrogen. The suspension was partitioned between ethyl acetate (30 mL) and 5% aqueous sodium bicarbonate (20 mL). The ethyl acetate layer

was washed with brine (15 mL), dried with magnesium sulfate, filtered and evaporated to give the title compound as an oil (350 mg), which contained residual tetrahydrofuran as observed by NMR. The foam was purified by preparative TLC plates (3x1000 micron, eluted with 10% ethyl acetate) to give the title compound as a foam (278 mg). -H NMR (CDC1 ₃ , 400 MHz) δ 0.61 (q, Si(CH2 i3)3, 0.94 (t, Si(CH2CH ₃ ) ₃ , 1.13 (d, I-CH ₃ ), 1.23 (d, CH3CHOH), 3.10 (t, CH ₂ Ar), 3.29 (dd, H-6), 3.63 (dq, H-l), 3.93 (t, CH ₂ OH), 4.27 (p, CH ₃ CHOH), 4.33 (dd, H-5), 5.30 and 5.50 (two d's, 7.00 and 7.28 (d, 2ArH), 7.49 (s, ArH), 7.68 and 8.23 (two d, CH ₂ C ₆ N0 ₂ ).

Step 2 p-Nitrobenzyl (lR.5S.6S)-2-r4-(5-(trifluoromethanesulfonatoethyPthien- 2-vPthiazolo-2-thiol-6-r(lR)-triethylsilyloxyeth-l-yll-l -methylcarbapen- 2-em-3-carboxylate

A solution of p-Nitrobenzyl (lR,5S,6S)-2-[4-(5-(hydroxy- ethyl)thien-2-yl)thiazolo-2-thio]-6-[( 1 R)-triethylsilyloxyeth- 1 -yl]- 1 - methylcarbapen-2-em-3-carboxylate (278 mg, 0.396 mmol) and 2,6- lutidine (0.11 mL, lmmol) in methylene chloride (5 mL) was cooled in an ice bath under a nitrogen atmosphere. Trifluoromethanesulfonic anhydride (0.084 mL, 0.5mmol) was added dropwise over 1 minute. After 40 minutes, the mixture was partitioned between methylene chloride (6 mL) and water (6 mL). The aqueous layer was re-extracted with methylene chloride (2 x 10 mL) and the combined methylene chloride layers were dried with magnesium sulfate, filtered and evaporated to give the title compound as an oil (318 mg). ⁱ H NMR (CDCI ₃ , 500 MHz) δ 0.61 (q, Si(CH2_CH ₃ ) ₃ , 0.95 (t, Si(£H2CH ₃ ) ₃ , 1.14 (d, I-CH ₃ ), 1.24 (d, CH3CHOH), 3.29 (dd, H-6), 3.37 (t, CH ₂ Ar), 3.69 (dq, H-l ), 4.28 (p, CH ₃ CHOH), 4.33 (dd, H-5), 4.73 (s, CH ₂ OTf), 5.32 and 5.50 (two d's, CH ₂ C ₆ H ₄ N0 ₂ ), 6.91 and 7.30 (d, 2ArH), 7.39 (s, ArH), 7.68 and 8.24 (two m's, CH^^NO; _? ).

Step3 p-Nitrobenzyl ( 1 R.5S.6SV2- (4-((5-carbamoylmethyl- 1.4- diazabicyclof2.2.21oct-l -yPethyPthien-2-yPthiazolo-2-thio1-

6-11 (R)-triethylsilyloxyeth- 1 -yll- 1 -methylcarbapen-2-em 3-carboxylate bis trifluoromethanesulfonate

A solution of p-Nitrobenzyl (lR,5S,6S)-2-[4-(5-(trifluoro- methanesulfonatoethy l)thien-2-y l)thiazolo-2-thio] -6- [( 1 R)-triethy 1- silyloxyeth- 1 -yl]- 1 -methylcarbapen-2-em-3-carboxylate (318 mg, 0.39 mmol) and carbamoylmethyl- l,4-diazoniabicyclo[2.2.2]octane bis trifluoromethanesulfonate (120 mg, 0.374 mmol) in acetonitrile (2 mL) was stirred for 1 hour at room temperature. The solution was evaporated and the residue was triturated with diethyl ether (10 mL). The resulting precipitate was filtered and provided the title compound as an amorphous solid (385 mg).

Step 4

(lR.5S.6SV2-l(4-((5-carbamoylmethyl-1.4-diazabicvclor2.2. 21oct-l- vPethyPthien-2-vPthiazolo-2-thio1-6-ri(RVhvdroxyethyll-l- methylcarbapen-2-em 3-carboxylate chloride

The pH of a solution of p-Nitrobenzyl (lR,5S,6S)-2- [(4-((5-carbamoylmethyl-l,4-diazabicyclo[2.2.2]oct-l-yl)ethy l)thien-2- yl)thiazolo-2-thio]-6-[ 1 (R)-triethylsilyloxyeth- 1 -yl]- 1 -methylcarbapen- 2-em 3-carboxylate bis trifluoromethanesulfonate (380 mg, 0.331 mmol) in a mixture of tetrahydrofuran (8 mL) and water (4 mL), was adjusted to 2.40 with IN trifluoromethanesulfonic acid. After stirring for 35 minutes at room temperature, the pH was adjusted to 6.0 with 5% aqueous sodium bicarbonate. Butanol (10 mL), ethyl acetate (6 mL), and 0.1N pH7 sodium phosphate buffer (22 mL) were added and the rapidly stirred mixture was hydrogenated (atmospheric pressure) in the presence of 10% palladium on carbon (120 mg). After 2 hours, the mixture was filtered through a 0.45 micron acrodisc, the aqueous layer was removed and the organic layer was extracted with water (1x10 mL). The combined aqueous layers were sparged with nitrogen and the aqueous layer was loaded onto a weak cation exchange column (16 mL of Bio-rad macroprep cm support). The column was eluted with water (9x 16 mL) and then with 5% aqueous sodium chloride. Fractions (15 mL each)

were collected and the product eluted in fractions 2-5 of the 5% aqueous sodium chloride eluent. The combined fractions were loaded onto an amberchrom column (8mL), and the column was washed with water (50mL) and then eluted with 20% isopropanol/ water collecting 5 mL fractions. Fractions 1-3 were combined and evaporated to ca. ImL and lyophilized to give the title compound as a white solid (50 mg). UV (water) λ _max 295nm (ε 21,650).

-H NMR (D ₂ 0, 500 MHz) δ 0.86 (d, 1-CH ₃ ), 1.13 (d, CH3CHOH), 3.31 (dq, H-l), 3.45 (dd, H-6), 3.42 (t, CH ₂ Ar), 3.93 (CH ₂ N), 4.18 and 4.75 (m, NCH ₂ CH ₂ N), 4.10 (dd, H-5), 4.18 (p, CH ₃ CHOH), 4.44 (s, CH ₂ C(0)NH2), 6.95 and 7.20 (d, ArH) and 7.55 (s, ArH).

EXAMPLE 16

(lR.5S.6SV2-r(4-((4-(CARBAMOYLMETHYL-1.4- PI AZABICYCLOr2.2.21QCT- 1 - YL^ETHYL^THIOPHEN-2-

YLyiΗIAZOLO-2-THIQl-6-r 1 ( R VHYDROXYETHYL1- 1 -

METHYLCARBAPEN-2-EM 3-CARBOXYLATE CHLORIDE

Step l p-Nitrobenzyl (1R.5S.6S 2-r4-r4-(hvdroxyethvPthiophen-2-vPthiazolo- 2-thiol-6-IY 1 R Vtriethylsilyloxyeth- 1 -yll- 1 -methylcarbapen-2-em-3- carboxvlate

A mixture of 4-(4-(hydroxyethyl)thiophen-2-yl)-2- mercaptothiazole (150 mg, 0.617 mmol), p-Nitrobenzyl (1R,5R,6S)- 2-(trifluoromethanesulfono)oxy-6-[(lR)-triethylsilyloxyeth-l -yl]- l-methylcarbapen-2-em-3-carboxylate (375 mg, 0.617 mmol) and powdered lithium hydroxide monohydrate (32 mg, 0.77 lmmol) in tetrahydrofuran (4 mL) was stiπed at room temperature for 2 hours under nitrogen. The suspension was partitioned between methylene chloride (lOmL) and 5% aqueous sodium bicarbonate (10 mL). The methylene chloride layer was dried with magnesium sulfate, filtered and evaporated

to give the title compound as an oil (350mg), which contained residual tetrahydrofuran as observed by NMR. The foam was purified by preparative TLC plates (3x1000 micron, eluted with 10% ethyl acetate) to give the title compound as a foam (235 mg). -H NMR (CDC1 ₃ , 400 MHz) δ 0.59 (q, Si(CH2£H3) ₃ , 0.95 (t, Si(£H ₂ CH ₃ ) ₃ , 1.12 (d, I-CH ₃ ), 1.23 (d, CH ₃ CHOH), 2.89 (t, CH ₂ Ar), 3.28 (dd, H-6), 3.67 (dq, H-l), 3.89 (t, CH ₂ 0), 4.27 (p, CH ₃ CHOH), 4.32 (dd, H-5), 5.31 and 5.50 (two d's, CH ₂ C ₆ H ₄ N0 ₂ ), 7.04 and 7.40 (d, 2ArH), 7.49 (s, ArH), 7.67 and 8.23 (two d, CH2C ₆ H4NO2).

Step 2 p-Nitrobenzyl ( 1 R.5S.6S V2-r4-(4-(trifluoromethanesulfonatoethyP thiophen-2-vPthiazolo-2-thiol-6-[(lR)-triethylsilyloxyeth-l- yll-l- methylcarbapen-2-em-3-carboxylate

A solution of p-Nitrobenzyl (lR,5S,6S)-2-[4-(4-(hydroxy- ethyl)thiophen-2-yl)thiazolo-2-thio]-6-[(lR)-triethylsilylox yeth-l-yl]- l-methylcarbapen-2-em-3-carboxylate (235mg, 0.335mmol) and 2,6- lutidine (0.093 mL, 0.837mmol) in methylene chloride (5 mL) was cooled in an ice bath under a nitrogen atmosphere. Trifluoromethane¬ sulfonic anhydride (0.070mL, 0.419mmol) was added dropwise over 1 minute. After 45 minutes, the mixture was partitioned between methylene chloride (6 mL) and 0.5N hydrochloric acid (10 mL). The methylene chloride was dried with magnesium sulfate, filtered and evaporated to give the title compound as an oil (260 mg). ⁱ H NMR (CDCI ₃ , 500 MHz) δθ.60 (q, Si(CH ₂ CH ₃ ) ₃ , 0.95 (t, Si(CH ₂ CH ₃ ) , 1.14 (d, I-CH ₃ ), 1.24 (d, CH ₃ CHOH), 3.18 (t, CH ₂ Ar), 3.30 (dd, H-6), 3.69 (dq, H-l), 4.28 (p, CH ₃ CHOH), 4.33 (dd, H-5), 4.73 (s, CH ₂ OTf), 5.32 and 5.50 (two d's, CH ₂ C ₆ H4NO ₂ ), 7.11 and 7.43 (d, ArH), 7.39 (s, ArH), 7.69 and 8.24 (two m's, 0^0^0 ₂ ).

Step3 p-Nitrobenzyl (lR.5S.6SV2-f(4-((4-carbamoylmethyl-l .4- diazoniabicyclor2.2.21oct-l -yPethyPthiophen-2-yPthiazolo-2-thiol-6- [ 1 (R)-triethylsilyloxyeth- 1 -yll- 1 -methylcarbapen-2-em 3-carboxylate bis trifluoromethanesulfonate

A solution of p-Nitrobenzyl (lR,5S,6S)-2-[4-(4-(trifluoro- methanesulfonatoethyl)thiophen-2-yl)thiazolo-2-thio]-6-[(lR) -triethyl- silyloxyeth-l-yl]-l-methylcarbapen-2-em-3 -carboxylate (260 mg, 0.3 lmmol) and carbamoylmethyl- l,4-diazoniabicyclo[2.2.2]octane bis trifluoromethanesulfonate (107 mg, 0.335 mmol) in acetonitrile (2 mL) was stirred for 5 hours at room temperature. The solution was evaporated and the residue was dried under vacuum to give the title compound as a glass (367 mg).

Step 4

(lR.5S.6S 2-r(4-((4-carbamoylmethyl-1.4-diazabicvclor2.2.21oct-l- vPethvPthiophen-2-vPthiazolo-2-thiol-6-l 1 (R hydroxyethyll- 1 - methylcarbapen-2-em 3-carboxylate chloride

The pH of a solution of p-Nitrobenzyl ( 1 R,5S,6S)-2-[(4- ((4-carbamoylmethyl- 1 ,4-diazabicyclo[2.2.2]oct- 1 -yl)ethyl)thiophen-2- yl)thiazolo-2-thio]-6-[ 1 (R)-triethylsilyloxyeth-l -yl]- 1 -methylcarbapen-2- em 3-carboxylate bis trifluoromethanesulfonate (367 mg, 0.3 lmmol) in a mixture of tetrahydrofuran (6 mL) and water (3 mL), was adjusted to 2.40 with IN trifluoromethanesulfonic acid. After stirring for 35 minutes at room temperature, the pH was adjusted to 6.0 with 5% aqueous sodium bicarbonate. Butanol (8 mL), ethyl acetate (5 mL), and 0.1N pH7 sodium phosphate buffer (15 mL) were added and the rapidly stiπed mixture was hydrogenated (atmospheric pressure) in the presence of 10% palladium on carbon (100 mg). After 2 hours, the mixture was filtered through a 0.45 micron acrodisc, the aqueous layer was removed and the organic layer was extracted with water (1x10 mL). The combined aqueous layers were sparged with nitrogen and the aqueous layer was loaded onto a weak cation exchange column (2x 19cm of Bio-rad macroprep cm support). The column was eluted with water (200 mL) and then with 5% aqueous sodium chloride. Fractions (8 mL each) were collected and the product eluted in fractions 5-13 of the 5% aqueous sodium chloride eluent. The combined fractions were loaded onto an amberchrom column (8 mL), and the column was washed with water (50 mL) and then eluted with 20% isopropanol/ water collecting 5 mL fractions. Fractions 1 -4

were combined and evaporated to ca. 1 mL and lyophilized to give the title compound as a white solid (60 mg).

UV (water) λmax 295 nm (ε 18,400).

-H NMR (D ₂ 0, 500 MHz) δ 0.86 (d, 1-CH ₃ ), 1.12 (d, CH3CHOH), 3.08

(dq, H-l), 3.23 (t, CH ₂ Ar), 3.29 (dd, H-6), 3.92 (t, CH ₂ N), 4.00 (dd, H-5),

4.08 (p, CH ₃ CHOH), 4.14 and 4.32 (m, NCH ₂ CH ₂ N), 4.44 (s,

CH ₂ C(0)NH2), 7.25 and 7.33 (s, ArH) and 7.61 (s, ArH).

EXAMPLE 17 Using the procedures set forth in Examples 1-6, the 2- (diphenylphosphono)oxy carbapenem intermediate A below is reacted with HSHet, with Het as defined in Table I, to produce Compound la. The values of R- , M and -Het of Compound la are set forth in Table I.

TABLE I

EXAMPLE 18 Using the procedures set forth in Examples 8-16, compound AL HSHet and Q* as set forth in Table II are reacted to produce a compound of formula ϋa in which -Het* is as defined in Table II.

TABLE π