FI78094C - Process for the preparation of antibiotically active carbapenem compounds - Google Patents

Description

1 78094

Method for preparing antibiotically active carbapenem derivatives

This invention relates to a process for the preparation of novel carbapenem-5 antibiotics of formula I.

R15 OH H g I ^ CHoCH --- S-A-C-N®-R5

10 3 1 I W

J-N- \ 2 U COOR ^ where c R is alkyl of 1 to 3 carbon atoms, carboxymethyl or allyl; A is straight or side chain C 1 -C 6 alkylene; R is hydrogen, an anionic charge or a common easily removable carboxyl protecting group, provided that when R is hydrogen or a protecting group, a counterion is also present; R * 5 is hydrogen or methyl; and -Οθ ~ 25 is a heteroaromatic group quaternized by R 1, which is a pyridinium, tetrazolium, imidazolium, triazolium, thiazolium or thiadiazolium group, and which may be substituted by 1 to 3 methyl, carboxymethyl or benzyl and has joined group A via a ring carbon atom. Numerous β-lactam derivatives with a carbapenem core have been described in the literature. These carbapenem derivatives have been reported to be useful as antibacterial agents and / or β-lactam. inhibitors.

The original carbapenem compounds were natural products such as thienamycin with the formula 5 ° H * '- | 1 — SCH2CH2NH2

10 -N-L

tr xCOOH

obtained by fermentation using Streptomyces cattleys (U.S. Patent 3,970,357). Thienamycin 15 is an exceptionally potent broad-spectrum antibiotic with remarkably potent activity against a variety of Pseudomo-nas species, organisms that have been remarkably resistant to β-lactam antibiotics.

Other natural products with a carbapenem core include olivic acid derivatives such as the antibiotic MM 13902 of the formula ch3 25 - / \ - SCH = CHNHCOCH o ho3so I il 0

CT N x COOH

30 described in U.S. Patent 4,113,856, an antibiotic MM 17880 having the formula ch 3 -S \ - SCH 2 CH 2 NHCOCH 2

J5 HO3SO Γ I M

“ΘΟΗ 3,78094 described in U.S. Patent 4,162,304, an antibiotic MM 4550A having the formula 5 CH3

I 3 O

S-CH = CHNHCOCH-, ho3s ° [JJ 3

0 XOOH

10 described in U.S. Patent 4,172,129 and the antibiotic 890Ag of the formula CH3 O ho3sc> I-f] - SCH = CHNHCCH3 cr N xooh 20 described in U.S. Patent 4,264,735.

The absolute configuration of naturally occurring thienamycin is 5R, 6S, 8R. This isomer, as well as the other seven isomers of thienamycin, can be obtained by the total synthesis disclosed in U.S. Patent 4,234,596. Methods for the total synthesis of thienamycin are also described, e.g., in U.S. Patent 4,287,123, 4,269,772, 4,282,148, 4,273. 709, 4,290,947 and European Patent Application 7973.

30 Due to the exceptional biological effect of thienamycin, a large number of derivatives have been synthetically prepared and described in the literature. Examples are compounds of the formula 78094 OR3 ^ —sch2ch2nr1r2 5 \ - n_y

v XCOOH

o 1

wherein R is aryl, alkyl, acyl or aralkyl and R and R 2 are independently members of the group consisting of H and acyl, including acyl of type O

11 1 1 1 -C-RA, wherein R may be, inter alia, alkyl substituted with a quaternary ammonium group, e.g.

And described in U.S. Patent 4,226,870, and compounds of the above formula wherein R 3 is H, acyl, or a monovalent optionally substituted hydrocarbon radical; R 1 is optionally substituted alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkenylalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl or heteroaralkyl, and R is acyl including acyl which is

II

is of the type -C-R, where R is alkyl having a quaternary ammonium group as a substituent, e.g.

And described in GB Patent 1,604,276 (see also U.S. Patent 4,235,917).

5,78094

Synthetically prepared derivatives of thienamycin have been further described e.g. GB Patent 1,604,275, U.S. Patent 4,235,920 and EP 40,408, 38,869, 24,832, 44,170 and 50,334.

Although, as indicated above, carbapenem derivatives having a 2-substituent which can be represented by the general formula -SA-Het, wherein A represents an alkylene group and Het represents a heterocyclic or heteroaromatic group, there has been no description of carbapenem having Het would be a radical of the formula - ^ \ © -R5, which is a characteristic structural feature of the carbapenems prepared according to this invention.

Despite the huge number of carbapenem derivatives described in the literature, there is still a need for new carbapenems that are more advantageous than known derivatives in terms of their spectrum of activity, efficacy, stability and / or toxic side effects.

The novel compounds of formula I prepared according to the present invention are effective antibacterial agents. They have been shown to have surprisingly low toxicity and a very broad spectrum of activity, as shown by the experimental results presented below.

As used in the definition of the compounds, the term "ordinary readily removable carboxyl protecting group" means a known ester group used to protect the carboxyl group during the chemical reaction steps described below and which can be removed, if desired, by methods that do not cause any significant molecular damage.

Decomposition of the remainder, e.g., by chemical or enzymatic hydrolysis, treatment with chemical reducing agents under mild conditions, irradiation with ultraviolet light, or catalytic hydrogenation. Examples of such ester protecting groups are benzhydryl, allyl, p-nitrobenzyl, 2-naphthylmethyl, benzyl, trichloroethyl, silyl such as trimethylsilyl, phenacyl, p-meotoxybenzyl, acetonyl, o-nitrobenzyl, 4-pyridylmethyl and C 1 -C 8 alkyl, such as ethyl 10 or t-butyl. Such protecting groups include groups which, by hydrolysis under physiological conditions such as pivaloyloxymethyl, acetoxymethyl, phthalidyl, indanyl and methoxymethyl. A particularly preferred carboxyl protecting group is p-nitrobenzyl, which is readily removed by catalytic hydrogenolysis.

The pharmaceutically acceptable salts mentioned above include non-toxic acid addition salts, e.g., salts with inorganic acids such as hydrogen chloride, hydrogen bromide, hydrogen iodide, phosphoric acid, sulfuric acid, etc., and salts with organic acids such as maleic acid, acetic acid, citric acid, succinic acid, succinic acid, succinic acid fumaric acid, mandelic acid, ascorbic acid, lactic

C

with acid, glucosic acid and malic acid. When the RJ group or the radical 25 O * 30 contains acidic groups, suitable base addition salts of these functional groups, e.g. metal salts (e.g. sodium, potassium, calcium and aluminum), ammonium salt and salts of non-toxic amines (e.g. with trialkylamines, procaine, dibenzyl-35-amine, 1-ephenamine, N-benzyl-N-phenethylamine, N, N'-dibenzylethyleneamine, etc.).

7 78094

In the compounds of formula I, the alkylene radical A may be straight-chain or side-chain and may have 1 to 6 carbon atoms. Preferably A is - (CH 2) n -, where n is 1 or 2 and particularly preferably A is 5. Particularly preferred are compounds of formula I having an absolute configuration of 5R, 6S, 8R.

The carbapenem derivatives of the formula I are prepared according to the invention in such a way that a) the compound of formula (II) 10 R15

? H H g 1 / N

CH 3 CH - A --- 11 15 -N- \ XOOR 2 wherein R 15 and A are as defined above and O is a heteroaromatic radical selected from pyridyl, tetrazolyl,

Imidazolyl, triazolyl, thiazolyl or thiadiazolyl and which may be substituted by 1-3 methyl, carboxymethyl or benzyl and which is attached to group A

n? via a ring carbon atom, and R is a common easily removable carboxyl protecting group, is reacted in an inert organic solvent with an alkylating agent of formula R5-X 'wherein R1 is as defined above and X' is a common leaving group such as halogen, so that a substituent on a compound of formula II

-O

35 8 78094 The ring type is quaternized with R5 to form a compound of formula (I ') R15

5 oh H s I

CHjCH - f / | TS — A Χ'θ cr N COOR2 '10 wherein R15, R2, A, —and X' are as defined above; and optionally replacing the counterion X 'with a different counterion and optionally removing the carboxyl protecting group 2 1 R to give the desired unprotected compound of formula I or a pharmaceutically acceptable salt thereof, or b) (1) a compound of formula (III) R 15

oh h a I

20 I I / V p

CH 3 CH - Y

1 111 cm 2 N 2 O 2 O 2 · 25 wherein RJ and R are as defined above are reacted in an inert organic solvent with a reagent capable of attaching a conventional leaving group L such as a diphenylphosphoryloxy group to the 2-position of a compound of formula III to give a compound of formula (IV) 30 R15? HH ιΛ,

C "3CH-f | T

\ 00R2 · 9 78094

o I

wherein L and are as defined above; (2) reacting the obtained compound of formula IV in an inert organic solvent and in the presence of a base with a mercaptan reagent of formula 5

HS-A- -O

wherein A and O are as defined above, and the resulting compound of formula II is converted as described in a) above to the desired compound of formula I or a pharmaceutically acceptable salt thereof, or c) a compound of formula (IV) 15 H f'5

OH H

CHoCH - Y

3 I | IV

--- N-k O ncoor ^ 20 ii IS.

wherein R *, RXJ and L are as defined above, is reacted with a thiol compound of formula (VII)

HS-A- £ - N®-R5 X® VII

wherein A and are as defined above and 5 ^ is the counter anion, in an inert solvent and in the presence of a base, to form a carbapenem product of formula (I ') R15 OH HHI / ""' N Λ 35 ch3ch - Γ | Γ ”δ— A - ^ _ y.N®-R5 x® ra N o »^ COOR ^ 10 78094 wherein R15, R2 ', A, (^ ^ J®-R5 and X® are as defined above and, if desired, the carboxyl protecting group R2 is removed to give the corresponding formula A deprotected compound or a pharmaceutically acceptable salt thereof.

In process variant b), the starting material III is reacted in an inert organic solvent such as methylene chloride, acetonitrile or dimethylformamide with about an equimolar amount of R 0 -L, such as p-toluenesulfonic anhydride, p-nitrobenzenesulfonic anhydride, p-nitrobenzenesulfonic anhydride isopropyylibentseenisulfonihappoanhydridin, methanesulfonic, trifluorimetaanisulfonihap-anhydride, diphenylchlorophosphate, toluenesulfonyl chloride, p-bromobenzenesulfonyl chloride or the like material, wherein L is the corresponding leaving group 15 such as toluenesulfonyloxy, p-nitrobenzenesulfonyl-oxy, difenoksifosfinyylioksi, and one of the other leaving groups, which are regularly used in conventional methods and are well known in the art. The reaction to secure the leaving group at the 2-position 20 of the starting material III is preferably carried out in the presence of a base such as diisopropylethylamine, triethylamine, 4-dimethylaminopyridine, or the like at a temperature of about -20 to + 40 ° C, most preferably about 0 ° C. The leaving group L of Intermediate IV may also be halogen, in which case such a group is placed by reacting Intermediate III with a halogenating agent such as O 3 ClCl 2 (oxalyl chloride or the like in a solvent such as CH 3 CN, THF or the like). in the presence of a base such as di-isopropylethylamine, triethylamine, 4-dimethylaminopyridine, etc. If desired, intermediate IV can be isolated, but is usually used in the next step without isolation or purification.

35 Intermediate IV is then converted to Intermediate II

according to the usual substitution reaction. Thus, intermediate 11 78094 IV can be reacted with approximately an equimolar amount of a heteroaralkyl mercaptan reagent of formula

O

c HS-A-μ- N

5 where A and

1. "O

are as defined above in an inert organic solvent such as dioxane, dimethylformamide, dimethylsulfoxide or acetonitrile and in the presence of a base such as diisopropylethylamine, triethylamine, sodium hydrogen carbonate, potassium carbonate or 4-dimethylaminopyridine. The temperature of the substitution reaction is not critical, but the preferred temperature range is from about -40 ° C to + 25 ° C. Most conveniently, the reaction is carried out using cooling, e.g., at about 0 ° C to -10 ° C.

The ring nitrogen quaternization of the heteroaralkyl group of Intermediate II is performed by reacting Intermediate II in an inert organic solvent with at least an equivalent amount (up to about 50% molar excess) of an alkylate of formula R5-X 'as defined above and X' is a common leaving group. such as halogen (chlorine, bromine or iodine, preferably iodine). Examples of suitable inert organic solvents are chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethyl sulfoxide and dimethylformamide. The temperature of the alkylation reaction is not critical and temperatures in the range of about 0 ° C to 35 ° C are preferred. Most conveniently, the reaction step is performed at room temperature.

12 78094

The resulting compound of formula 1 'has a counterion X' (e.g. a derivative of the alkylating agent used) associated therewith, and may be present at this stage or at a later stage, i. after the deprotection step, replace with a different counterion, e.g. a more pharmaceutically acceptable counterion, by conventional methods. Alternatively, my counterion can be removed later during the deprotection step.

The deprotection step to remove the carboxyl protecting group R of the compound 1 'is carried out by conventional methods such as solvolysis, chemical reduction or hydrogenation. When a protecting group such as p-nitrobenzyl, benzyl, benzhydryl or 2-naphthylmethyl which can be removed by catalytic hydrogenation is used, intermediate 1 'can be treated in a suitable solvent such as dioxane-water-ethanol, tetrahydrofuran-dipotassium hydrogen phosphate-isopropane-aqueous phosphate or in a similar solvent at a hydrogen pressure of 1 to 4 atmospheres, in the presence of a hydrogenation catalyst such as palladium on carbon, palladium hydroxide, platinum oxide or the like at a temperature of 0 to 50 ° C for about 0.24 to 4 hours. When R is a group such as o-nitrobenzyl, photolysis can also be used to deprotect. 2,2,2-trichloroethyl-like protecting groups can be removed using mild zinc reduction. The allyl protecting group can be removed with a catalyst comprising a mixture of a palladium compound and triphenylphosphine in an aprotic solvent such as tetrahydrofuran, diethyl ether or methylene chloride. Similarly, other common carboxyl protecting groups can be removed by methods known to those skilled in the art. Finally, as mentioned above, compounds of formula 1 'wherein R is a physiologically hydrolyzable ester such as acetoxymethyl, phthalidyl, indanyl, pivaloyloxymethyl, methoxymethyl-35, etc. may be administered directly to a patient without deprotection upon hydrolysis of such esters in vivo under physiological conditions.

i3 7 8094

In a variation of the above process, the carboxyl protecting group of Intermediate II may be removed prior to the quaternization step. Thus, the carboxyl protecting group is removed as described above to give the corresponding free carboxylic acid and the free acid is then quaternized with the alkylating agent R5-X 'to give the desired quaternized product of formula I. In the deprotection of deprotected intermediate II, the solvent may be water or an unreacted organic solvent, or a mixture thereof. Examples of suitable solvents are water, organic solvents such as chloroform, methylene chloride, tetrahydrofuran, dioxane, acetone, dimethyl sulfoxide and dimethylformamide, and mixtures of water and an organic solvent such as mixtures of water-acetone or water-dimethylformamide. The quaternization temperature of intermediate Ha is then not critical and temperatures in the range of about -40 ° C to about room temperature are convenient to use. Most preferably, the reaction is performed at about 0 ° C.

When the unprotected intermediate II is obtained as the carboxylate salt, it is desirable to add a strong acid such as toluenesulfonic acid to form the free carboxylic acid prior to quaternization. This has been found to greatly facilitate primary quaternization of ring nitrogen.

The process variation described above is particularly useful when the carboxyl protecting group is more easily removed from the non-quaternized intermediate II than from the quaternized compound I '. For example, in the preparation of a product of formula 30 9h3 0H H \ jG-v «Λ - SCH2 - \ _ / N ^ -coo® 35 14 78094 from an intermediate of formula

5, Λ-hr --O

Removal of the γ-n - oo coo 10 allyl protecting group prior to quaternization results in substantially better yields of the desired final product.

The starting material of formula IV used in process variant c) is described, for example, in European Patent Applications 38 869 and 54 917 and can be prepared by the general methods described therein. L represents a common leaving group such as chlorine, bromine, iodine, benzenesulfonyloxy, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy, methanesulfonyloxy, trifluoromethanesulfonyloxy, diphenoxyphosphinyloxy or di (trichloroethoxy) phosphinyloxy. The primary leaving group is diphenoxyphosphinyloxy. The preparation of compounds of formula IV is also described above in connection with process variant b).

Reaction of an intermediate of formula IV of formula VII

with a thiol compound according to the invention is carried out in an inert solvent such as acetonitrile, acetonitrile-dimethylformamide, tetrahydrofuran, tetrahydrofuran-H 2 O, acetonitrile-T 2 O or acetone in the presence of 30 bases. The nature of the base is not critical. Suitable bases include sodium hydroxide, diisopropylethylamine, 1,8-diazabicyclo [15.4] n-undec-7-ene, 1,5-diazabicyclo [3.3.3] n-non-5-ene and tri (C 1 -C 4) alkylamines such as triethylamine, tributylamine or tripropylamine. The reaction between intermediate IV and thiol VII can be carried out over a wide range of temperatures, e.g. -15 ° C to room temperature, but is preferably carried out at a temperature in the range of about -15 ° C to + 15 ° C, most preferably 0 ° C: around.

The carbapenem product prepared by the reaction of the quaternary amine thiol VII and intermediate IV is associated with a counter anion, e.g. (C 9 H 8 O 2) 2 ^ 2 ^ or an anion associated with a quaternary thiol, which can be replaced at this stage by conventional methods with a different anti-anion, e.g. an anion, which is more pharmaceutically acceptable. Alternatively, the counter anion can be removed later during the next deprotection step. When the quaternized carbapenem compound and the counter anion form an insoluble product, the product can be crystallized as it forms and recovered in pure form by filtration.

After formation of the desired carbapenem product, the carboxyl protecting group R of the compound 1 'can be removed, if desired, by conventional methods described above.

Thiol intermediates of formula VII can be prepared, for example, from the corresponding thiol acetate compound of formula

ch3cs-a — u N

25 where A and “Ό are as defined above. The thiolacetate compound is quaternized by reacting it in an inert organic solvent such as diethyl ether, dichloromethane, methylene chloride, dioxane, benzene, xylene, ie 78094 toluene or mixtures thereof with a suitable alkylating agent of formula R 5 -x defined and X 'is a common leaving group such as halogen (chlorine, bromine or iodine, most preferably iodine) or a sulfonate ester group such as mesylate, tosylate or triflate. The temperature of the alkylation reaction is not critical, and temperatures in the range of about 0 ° C to 40 ° C are preferred.

Prior to reaction with the carbapenem intermediate IV, the quaternized thiol acetate compound is treated with hydrolysis with an acid or base to prepare the quaternary thiol intermediate VII. This hydrolysis is performed primarily just prior to incorporation into compound IV to minimize the decomposition of the relatively unstable quaternary thiol compound VII.

By appropriately selecting the solvents, the reaction of intermediate III to the final product I can be carried out without isolating the various intermediates, i.e. using the "one pot" method. An example of such a method is illustrated in the following Example 22.

As with other β-lactam antibiotics, the compounds of general formula I can be converted by known methods into pharmaceutically acceptable salts, which are the main ones for the purposes of this invention! equivalent to unsalted compounds. Thus

, O

for example, a compound of formula I wherein R * is an anionic charge may be dissolved in a suitable inert solvent and then an equivalent amount of a pharmaceutically acceptable acid is added. The desired acid addition salt can be recovered by conventional methods, e.g., by solvent precipitation, freeze-drying, etc. When the compound of formula I has other basic or acidic reactive groups, pharmaceutically acceptable base addition salts and acid additions can be prepared in a similar manner by known methods.

i7 78094

It will be appreciated that certain products within the scope of formula I may be formed as optical isomers as well as their epimeric sexes. It is intended to include all such optical isomers and epimeric mixtures within the scope of this invention. For example, hydroxyethyl having 6 substituents may be in either the R or S configuration and the resulting isomers as well as their epimeric mixtures are included within the scope of this invention.

O

A compound of formula I wherein R * is hydrogen or an anionic charge, or a pharmaceutically acceptable salt thereof, may also be converted by conventional methods to the corresponding compound wherein R is a physiologically hydrolysable ester group, or a compound of formula I wherein R is a common carboxyl protecting group, can be converted to the corresponding compound in which R * is hydrogen, an anionic charge or a physiologically hydrolyzable ester group, or a pharmaceutically acceptable salt thereof.

The novel carbapenem derivatives of the general formula I in which R is hydrogen, an anionic charge or a physiologically hydrolyzable carboxyl protecting group, or their pharmaceutically acceptable salts, are effective antibiotics which act against various gram-positive and gram-negative bacteria. and can be used, e.g., as animal feed additives to promote growth, as nutrient preservatives, as bactericides in industrial applications, e.g. . However, they are particularly useful in the treatment of infectious diseases in humans and other living organisms caused by gram-positive or gram-negative bacteria.

These pharmaceutically active compounds can be used alone or in admixture as pharmaceutical compositions 78094 containing, in addition to the active carbapenem ingredient, a pharmaceutically acceptable carrier or diluent. The compounds can be administered in a variety of ways; essentially interesting routes are: oral, topical or parenteral (e.g. intravenous or intramuscular injection). The pharmaceutical combinations may be in solid form such as capsules, tablets, powders, etc., or in liquid form such as solutions, suspensions, or emulsions. Injectable compositions, the preferred route of administration, may be presented in unit dosage form in ampoules or in multi-dose containers, and may contain formulatory agents such as suspending, stabilizing and dispersing agents. The compositions may be in ready-to-use form or in powder form, which may be reconstituted with a suitable vehicle, such as sterile water, for administration at a time. The dosage to be administered depends to a large extent on the particular compound used, the particular mixture being respected, the route of administration, the vital functions and condition of the patient, and the particular site and body being treated. The choice of a particular primary dosage and route of administration is thus left to the discretion of the treating person. In general, however, the compounds can be administered parenterally or orally to mammalian patients in an amount of about 5 to 200 mg / kg / day. Administration is usually in divided doses, e.g., three to four times during the day.

To illustrate the potent broad-spectrum antibacterial activity of the carbapenems of this invention, both in vitro and in vivo, and the low toxicity of the compounds, the biological values for the currently preferred carbapenem compounds of the present invention are set forth below.

In vitro activity

The carbapenem compound samples prepared in Examples 1-2, after dissolving in water and diluting with nutrient broth, were found to give the following minimum inhibitory concentrations (MICs) in yug / ml against said 9 78094 and microorganisms as determined by incubation overnight at 37 ° C. In C by the tube dilution method.

N-formimidoylthienamycin is included in the following tables as a reference compound.

20 7 8 0 9 4

In vitro

Antibacterial activity of the carbapenem derivative of Example 1__ MIC (mcq / ml) 5 Organism New compound N-formimidoyl-thienamycin_ S. pneumoniae A-9585 0.25 0.004 S.pyogenes A-9604 0.06 0.001 S. aureus A- 9537 0.13 0.004 10 S. aureus + 50% Serum A-9537 0.03 0.016 S. aureus (Pen-res.) A-9606 0.06 0.008 S. aureus 15 (Meth-res.) A15097 4 0, 5 S. faecalis A20688 0.5 0.5 E. coli (10-4 dii.) A15119 0.06 0.016 E. coli 20 (ΙΟ-3) A15119 - 0.03 E. coli (ΙΟ-2) A15119 - 0.06 E. coli (ΙΟ "4) A20341-1 0.03 0.03 25 E. coli (ΙΟ-3) A20341-1 - 0.03 E. coli (ΙΟ-2) A20341-1 - 0, 13 21 78094

In vitro antibacterial activity of the carbapenem derivative of Example 1 - continued MIC (mcg / ml) N-formimidoyl 5 Organism New compound thienamycin No. K. pneumoniae A-9664 0.25 ^ t3-3 K. pneumoniae A20468 0.25 0 ^ 06 P. mirabilis A-9900 ° '13 ° t06 10 P. vulgaris A21559 0, 13 0 ^ 03 P. morganii A15153 0,13 0T13 P. rettgerx A22424 0,25 0 ^ 25 S. marcescens A20019 0,13 0, 03 S. cloacae A-9569 0.13 0.06 15 E. cloacae A-9656 0.13 0.06 P. aeruginosa A-9843A * ^?. aeruginosa A21213 ^ 0.25 H. influenzae A-9833 16 16 H. influenzae A20178 32 32 20 H. influenzae A21518 16 32 H. influenzae A21522 8 32 B. fraqilis A22862 °) 03 0.016 3. fraqilis A22053 ° t03 ° '06 B. fraqilis A22696 ° '25 ° T13 25 B. fraqilis A22863 ° T ° 3 1 22 78094

In vitro antibacterial activity of the carbapenem derivative of Example 2 MIC (mcg / ml)

Organism New compound N-formylddoyl-thiena- _mycin_ S. pneumoniae A-9585 0f001 0.002 S. pyogenes A-9604 0.001 0.002 S. aureus A-9537 0.004 0.004 10 S. aureus A-9537 0.016 0.016 + 50% Serum S. aureus A-9606 0.008 0.008 (Pen-res.) S. aureus A 15097 8 4 (Meth.-res.) S. faecalis A 20688 0.25 0.5 E. coli A 15119 0.016 0.016 E. coli A 20341-1 0.016 0.03 K. pneumoniae A9664 0.06 0.06 2q K. pneumoniae -A20468 0.13 0.13 P. mirabilis A9900 0.03 0.06 P, vulgaris A21559 0.016 0.03 P. moroanii A15153 0r06 0 , 13 P. rettoeri A22424 0.13 0.13 25 S. marcescens A20019 0.03 0.03 E. cloacae A9659 0.13 0.06 E. cloacae A9656 0.25 0.06 P. aeruginosa A9843A 8 1 p aeruginosa A21213 2 0.25 23 78094

In vivo activity

The in vivo therapeutic efficacy of the compound of Example 1 and N-formimidoyl-thienamycin after intramuscular administration to mice infected with various organisms for the experiment are shown in the following table. It indicates the PDc value (dose in mg / kg required to protect 50% of infected mice).

Protective effect in intramuscularly treated infected mice

Altisuts PD5Q / treatment (mgAg) (number of organisms) The N-formimidoyl

Organic compound Thienamycin 15 P. mirabilis A-9900 3.6 X 108 3.3 3/15 4 1 P. aeruginosa A-9843a 5.5 X 10 0.3 0.5 P. aeruginosa A-20481 5.4 X 10 ^ 0.63 0.4 P. aeruginosa A-20599 1.4 X 10 8 0.7 0.18 S. aureus A-9606 6.6 X 10® 0.09 0.07 20 s faecalis A- 20688 2.3 X 108 3.3 2.81 E. coli A-15119 6.2 X 106 0.6 2.51 K. pneumoniae A-9964 5.1 X 108 2.5 2.2

Actual Values 25 Treatment Schedule: With the exception of E. coli A15119 and K. pneumoniae A9964, mice were treated i.m. compounds 0 and 2 hours after infection. For E. coli and K. pneumoniae, the time of treatment was 30 and 3.5 hours after infection. In each experiment, 5 mice were used per dose.

24 78094

toxicity

The toxicity of the compound of Example 1 after intracranial administration to mice has been determined and is shown in the following table.

5 Toxicity to mice after intracranial administration_ * LD, -q Maximum dose (mg / kg), ,, i clinical toxicity-

Compound without img / signs

Compound of Example 1 14 5 N-formimidoylthienamycin 32 5 * - mean 25 / mouse / compound

Blood concentration in mice 15 after intramuscular administration

The blood concentrations and half-life of the compound of Example 1 in mice after intramuscular administration of 20 mg / kg are shown in the table below.

20 Blood concentration (μg / ml)

Compound 10 20 30 45 60 90 * teaspoon / 2 minutes after administration (min.) (/Ug.h/ml)

Compound 14 of Example 1 10.8 6.8 2.6 0.8 <0.6 10 6.3 2 5 N-fomimidoylthienamycin 12.6 9.9 7.3 2.6 2.6 0.7 <0.3_9_6

The compounds were dissolved in 0.1 mol. in phosphate buffer pH 7. The values are the values of one experiment; 4 mice were used per compound.

30 * teaspoon / 2 means half-life in minutes.

** AUC means the area under the curve.

25 '78094

Urinary Recovery The urinary recovery of the compound of Example 1 after intramuscular administration (20 mg / kg) to mice is shown in the following table.

5 Amount recovered from urine

Intramuscular dose to mice 20 mg / kg._

Percentage recovery from dose 10 0-3 3-6 6-24 0-24

Compound one hour after dosing

Compound of Example 1 26.1 0.5 0.1 37.7 + 6.7 N-formimidoylthienamycin 12.1 0.1 <0.1 12.2 + 3.6 The compounds were dissolved in 0.1 mol. . phosphate buffer pH 7. The values are the values of one experiment; 4 mice per compound.

Other Biological Values In Vitro Activity 20 The carbapenem compound samples listed below (identified by Example Number), after dissolution in water and dilution in broth, were found to show the following minimum inhibitory concentrations (MICs) in units / ug / ml of said microorganisms. as determined by incubation overnight at 37 ° C as a tube dilution. N-formimidoyl-thienazine is added to the following table as a reference compound.

26 78094 _MIC (/ uq / ml) _

Body Example 3 Example / 4 Example 5 Example 6 Example 7 MK 07871 £ i. pneuaoniae A-9585 0.002 0.002 0.00 «0.004 0.00« 0.002 L. pyogenea A-9604 0.002 0.001 0.00 «0.00« 0.00 «0.002 S. faecalis A20688 0.5 0.5 0.25 0.5 0.13 0.5 L. aureua A-9537 0.016 0.008 0.004 0.03 0.008 0.00 «L. aureua A-9S37 0.016 0.03 0.016 0.06 0.03 0.016 + S0! Serum S. aureua A-9606 0.016 0.016 0.008 0.03 0.016 0.008 (Pen-rea) aureua A1S097 4 4 8 4 2 4 (Meth-rea) L. eoli A15119 0.03 0.016 0.016 0.06 0.00 «0.016 L ·. coli A20341-1 0.016 0.016 0.016 0.06 0.004 0.03 X. pneumoniae A-9664 0.06 0.03 0.03 0.13 0.016 0.06 X. pneumoniae A20468 0.06 0 .03 0.06 0.25 0.016 0.13 E. cloacae A-9659 0.13 0.03 0.06 0.25 0.016 0.06 E. cloacae A-9656 0.13 0.06 0.13 0.25 0.016 0, 06 L. mrabllia A-9900 0.13 0.06 0.03 0.025 0.016 0.06 ^ vulcari1 A21559 0.016 0.016 0.016 0.06 0.008 0.03 iL °° rcanii A15153 0.13 0.016 0.06 0.13 0.016 0.13 P. rettgeri A22424 0.25 0.13 0.13 0.25 0.06 0.13 S. narceacens Λ20019 0.016 0.03 0.06 0.13 0.008 0.03 aerucinoaa A-9843a 22 4 8 1 0.25 aeruginoe A21213 0.5 0.13 2 1 0.25 0.25 H. influenzae A-9833 2 2 4> 32> 32 16 H. influenzae A21518 224> 32> 32 16 L. ragilia A22862 0 .25 0.06 0.03 0.03 0.016 0.06 L. iracilia A22696 0.5 0.5 0.25 0.25 0.25 0.13 L, N-farmimidoyl-thienamycin 78094 ----- MIC (jjcf / ml) _

Organism ^ - ^ - 8 9 Example 10 Example 11 MK 08871 £ neujnoiuae A-9585 0.002 0.001 01001 "7 ^" 5 ^ 5-

Lmi ^ A'9604 0.002 0.001 0.001 o 002 0.002 S ^ Uecali, A206B8 0.5 0.13 0 25 0 5 0.25 “ureus A-9537 0.008 0.00. 0 008 0004 0.002 S ^ iureu, a-9537 o, 03 0.008 0/0 <0.016 0.016 + S0% Serum 1. '' iiiureu, A-9606 0> 03 0.008 0.008 0.016 0.008 (P «nr« ») '' 'L ·« ureus A1509 7 - (Heth-res) L · £ 2li A15119 0.016 0.008 0 »16 0.016 0.016 - S2lL A20341-1 0.03 0.004 0.008 0.03 0.016 - P "Sy> oni« e A-9664 0.03 0.016 0.06 0.03 0.03 L · pneumoniae A20468 0.13 0.03 0 (13 0 , 13 o, 13

Ej cloacae 1-9659 0.13 0.03 o, 13 0 | 0ί 0.13 - Ξΐ21 £££. A-9656 0.06 0.03 0.13 0fl3 0.06

Li. nu_rabilis A-9900 0.13 0.016 0.11 0.03 0.03

Pj, vul A2 «A2II59 0.03 0.008 0.016 0.03 0.016 iL” ° rcanii A15153 0.13 0.03 0fX3 0.06 0.06

Pj reto A22424 0.13 0.06 0.13 0.13 0.13 S_. mareeseens A20019 0.06 0.016 0.06 0.06 0.03

Li aeruginosa A-984JA 1 2 32 0.5 1

Pj aeruginosa A21215 0.25 0.13 3 0.13 0.13 N-formimidylc-i-thienamycin 28 78094 ____MIC (pg / ml) _

Organism Example 12 Example 13 Example 14 MK 07871

Su. pneumonia A-958S 0 ^ 02 0.0005 0.0005 0.002 pvogenes Λ-9604 0.004 0.0005 0.0005 0.002! L 1 «· Cali» A20688 0.5 0.13 0.13 0.25 S. aureu » A-9537 0.008 0.008 0.008 0.002 aura A-9537 0.016 0.016 0.03 0.008 ♦ SOI serua aureu »A-9606 0.03 0.008 0.016 0.008 (Pen-rae) S. aureu» A1S097 - (Math-ra ») E. coli A15119 0.016 0.004 0.008 0.016 E. coli A20341-1 0.008 0.008 0.008 0.016 K. oneunoniae A-9664 0.03 0.03 0.03 0.03 K. pneunoniae A20468 0.06 0.13 0.03 0.06

Ei cloacae A-9659 0.06 0.06 0.03 0.06 E. cloacae A-96S6 0.03 0.03 0.03 0.06 P. mirabill »A-9900 0.03 0.016 0.016 0.016 £ i vulgari »A21559 0.016 0.008 0.016 0.016 M. morcanii A15153 0.06 0.016 0.03 0.06 P. rettcerl A22424 0.13 0.25 0.06 0 ^ 13 S. ffiarcescens A20019 0.03 0.016 0.016 0.03 P. aeruginosa A-9S43A 16 32 8 1 P. aeruginosa A21213 2 2 0.5 0.13 N-formimidoyl-thienamycin

Organism Ex.15 "A" Ex.15 "B" Ex.15 "C" MK 0787 * 29 78094 _MIC tyjg / ml) S · pneumoniae Λ-9585 0.0005 0.002 0.0005 0.0005 S · pvonenes A- 9604 0.0005 0.002 0.001 0.0003 li - «ecalis A2068B 0.13 0.25 0.5 0.5 aureu» A-9537 0.03 0.004 0j004 0.016 aureu »A-9537 0.03 0.008 0.016 0.06 * 501 »erum S ^ _ aureu» A-9606 0.004 0.008 0.008 0.03 CPen-re ») S. aureu» A15097 - - - - (Meth-re1)

Ei coli A15119 0.004 0.008 0.008 0.06 lu. ° C A20341-1 0.004 0.016 0.008 0.03 K. pneumoniae A-9664 0.008 0.03 0.03 0.06 K. pneumoniae A2046B 0.008 0.06 0.016 0.13 E. cloacae A-9659 0.016 0.13 0.016 0, 13 S. cloacae A-9656 0.016 0.13 0.03 ° | 13 P. rviraSili »A-9900 0.008 0.06 0.03 0.06 P. vMlaari» A21559 0.008 0.016 0.008 0.06 M. morcanli A15153 ° i03 ° i13 ° i ° 1 ° l25 L. rpppgari A22424 0.03 ° i13 ° 113 ° »25 S _; _ aarcescens A20019 0.008 0.016 0.016 0.13

Li ** ru ° ino »» A-9843A ° 1S 2 ®

P. aeruginosa A21213 0.03 ^ 0.13 0.13 ° 4S

N-fonrdJnidoyyli-täenainysiini

Organism_ Eg 16 Example 17 MK 0787 30 7 8 0 9 4 S. pneumoniae A-9585 0.002 0.016 0.001

Si pyooenes A-9604 0.002 0.016 0.001 li faecalli A20688 1 4 0.25 auraus A-9537 0.006 0 ^ 25 0.001 S. aureus A-9537 0.03 1 0.008 - 50% Serum S-aureus A-9606 0.016 0.5 0 ^ 002 (Pen-res) 5i aureus A15097 1 IKeth-res) £ i cc li _ A15119 0.016 0.6 0.008 coli A20341-1 0.016 0.6 0.008 * · oneumonia »A-9664 0.06 0.13 0 , 03 * · pneumonia A20468 0.06 0.5 0.06 E. cloacae A-9659 0.06 2 0.06 E. cloacae A-96S6 0.06 2 0.06 P sirabills A-9900 0.06 0 , 13 0.016 P- vulcaris A21559 0.03 0.13 0.008 M · aoranii A15153 0.13 0, S 0.06 P. rettoeri A22424 0j25 2 0.06 S. ir.arcescens A20019 0.06 0.13 0.016

Pj_ aerucinosa A-9843A 0.25> 63 0.5 P aerucinosa A21213 0.13 16 0.13 N-formimidoyl-thienamycin

Organism Example 18 Example 19 Example 20 * MK 0787 31 78094 ii pneunoniae A-9585 1 0.002 0.0 «0.001 L. Pitäne» A-9604 2 0.002 0.13 0 ^ 002 ii taecalis A20688 «3 0.5 16 0 25 L. aureu »A-9S37 32 0,004 0,5 0T002 L. 1ure <i» A-9537> «3 0,008 2 0,004 ♦ 504 Sarum S. aureu» A-9606 »125 0.01« »125 0.004 (Pen -re ») S. aureus A15097 - - (Meth-r« 1) £; _ eoli A1S119 1 «0f00B 1 0j01« E ;, coli A20341-1 11 0,008 2 0,01 «K-pneumoniae A-9« « 4 32 0 ^ 03 4 o, 03 pneumoniae A20468 13 0,0 «4 0,0« £ i cloacae A-9659 13 0,03 8 0,0 «E. cloacae λ-9656 125 0,03 1« 0, 0 «P. mirapjli» A-9900 52 0 ^ 03 4 0,016 P. vulcari »A21S59 32 0,01« 4 0,01 «aoranali A15153 32 0T06 8 0,0« L. rattaeri A22424 32 0,13 8 0, 13 S. narcaican »A20019 32 0.03 4 0.03 P. aeruclno» a A-9843A 13 0.5 32 0.5 fi aeruginosa A21213 «3 0.0« 1 «0 ^ 13 ^ formimidoyl-thienamycin s2 78094

In vivo activity

The in vivo therapeutic efficacy of certain compounds of this invention and N-formimidoyl-thienamycin (MK0787) after intramuscular administration to mice infected with various organisms for the experiment is shown below. In this case, the dose of PD ^ Q (dose mg / kg) required to protect 50% of infected mice is reported.

Protective effect in intramuscularly treated IQ-infected mice ____ PD ^ / treatment (mg / kg)

Example Example Example Example Example Example

6 8 9 12 14 15 (“A”) MK 0787 S. property A9609 * 0 ^ 21 - 0.1 »0.07 - - ° · * 07 15 L £ 2ϋ A15119 - 0.86 lt2 - - - 3 pneumonia A9664 - 1.9 1.9 - *. ~ “3 9. miraSilie A9900 - 1.4 7.1 _ -. - 9 aeruginoas A9843A 0.4 0.19 0.19 1.9 0 ^ 45 0.3 »0.99 1 pp teruginoe A24091 * 0.33 0.19 '

Ex. Ex. Ex.

20 _3_4_7 MK 0787 S. aureua A9606 0.07 0.1 0.2 0.07 E. eoli A1S119 1 0.4 0.2 3 X. pneumonia Λ9664 331 3 P. Mirabilia A9900 2 «2, <» P. aeruginoat A9943A 0.5 0j2 0.2 0.5 P. aeruginoat A24081 0.8 0.2 0.1 0 ^ 4 25

E.g.

_5_MK 0787 S. auraus »f3 ° ’07 E. eoli * 2> 2 K. pneumoniae 3 273 30 pp. Mirabili. 10 * ». aaruoinoat A9 84 3A '' '33 7 8 0 9 4

Concentrations in the Blood of Mice Following Intramuscular Administration The blood concentrations and half-lives of some compounds of this invention after intramuscular administration to mice at a dose of 20 mg / kg are shown below.

C 1 2AUC

max Tl / 2 (ug.h / ml)

Compound _ (min.) _

Example 1 14 10 6.3 10 Example 2 13.9 9 5.3

Example 3 14.5 10 6.9

Example 4 15.5 11 7.7

Example 5 - -

Example 6 17.7 9 8.5 15 Example 7 19.2 11 11.8

Example 8 18.8 11 10.5

Example 9 16.7 12 8.5

Example 10 20.1 11 9.5

Example 11 14.9 11 7.4 20 Example 13 14.8 11 6.4

Example 14 15.8 13 7.6

Example 15 "A" 16.7 12 9.5

Example 15 "B" 15.9 10 7.4

Example 15 "C" 15.1 10 7.3 25 MK 0787 14.6 10 6

Example 17 11 -8 3.4

Example 18 14.9 6 3.9

Example 19 27 16.7 15.1

Example 20 28.4 14 15.6 30 The compounds were dissolved in 0.1 mol. phosphate buffer, pH 7. Values are based on one experiment; 4 mice per compound.

* denotes the half-life in minutes.

35 AUC denotes the area under the blood concentration-time curve.

2

The following examples illustrate this invention.

34 78094

Example 1 1-Methyl-4- (2-carboxy-6H- [1 (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-3-thiomethane Preparation of style pyridinium hydroxide inner salt 5 A.

OH / -y.

^ "r ^^ | rSCHiwNe'CH3 / 7-N - ^ e 10 O co2ö

OH H

15 \ I -7

N CO 2 pNB

20 OH

Kis ^ sarf \ 1 | T \ = / s

CT CO, pNB

2 25 OH _

30 // N '^ CO-pNB

O Z

3 pNB = -CH 2 - ^ 2 -nO 2 i.

35 7 8 0 9 4

A solution of 673 mg (1.86 mmol) of p-nitrobenzyl-ε-C 1- [1- (R) -hydroxyethyl] -3,7-dioxo-1-aza-bicyclo [3.2.07] hept-2 -ene-2-carboxylate (1) in 10 ml of acetonitrile, cooled to -10 ° C under a nitrogen atmosphere. A solution of 245 mg (1.90 mmol) of diisopropylethylamine in 1 mL of acetonitrile was added followed by dropwise addition of 510 mg (1.90 mmol) of diphenyl chlorophosphate in 1 mL of acetonitrile over 2 minutes. The resulting solution was stirred at -10 ° C for 15 minutes to give p-nitrobenzyl-3- (diphenylphosphoryloxy) -6β- [1- (R) -hydroxyethyl] -7-oxo-1- azabicyclo ^ 3,2, q / hept-2-ene-2-carboxylic azodicarboxylate. To this solution was added a solution of 245 mg (1.90 mmol) of diisopropylethylamine in 0.5 mL of acetonitrile followed by a solution of 270 mg (2.16 mmol) of 4-mercaptomethylpyridine in 0.5 mL. acetonitrile. The reaction mixture was stirred at -10 ° C for 60 minutes and the resulting white precipitate was collected by filtration and washed with 5 ml of ice-cold acetonitrile to give 660 mg (yield 76%) of compound 2 as white crystals, m.p. 145 ° C.

NMR (DMSO-d 6) 1.20 (3H, d, J = 6.0 Hz), 3.2-3.4 (3H, m), 3.7-4.1 (2H, m), 4, Δ (2H, s), 5.05 (1H, d, J = 4.0 Hz), 5.25 (1H, d, J = 14.0 Hz), 5.48 (1H, 25 d, J = 14.0 Hz), 7.40 (2H, d, J = 5.5 Hz), 7.70 (2H, d, J = 8.5 Hz), 8.23 (2H, d, J = 8, 5 Hz) and 8.58 (2H, d, J = 5.5 Hz).

IR (KBr) ν max: 3400, 1790, 1695 and 1600 cm-1. Analysis calculated for ^ 22Η21Ν3 ^ 6δ: 30 C, 58.01; H, 4.56; N, 9.23; S, 7.04.

Found: C, 57.74; H, 4.56; N, 9.58; S, 7.21.

To a solution of 660 mg (1.41 mmol) of compound in 2,140 mL of acetone was added 5 mL of methyl iodide. The reaction solution was stirred for 8 hours at 25 ° C.

The solvent was evaporated in vacuo to give a slightly yellowish solid which was triturated with diethyl ether to give 779 mg (90% yield) of the title compound 3 as a white amorphous solid, m.p. 130 ° C (decomp.).

NMR (DMSO-d 6) 1.15 (3H, d, J = 6.0 Hz), 3.2-3.4 δ (3H, m), 3.7-4.1 (2H, m), δ , 25 (3H, s), 4.30 (2H, s), 5.25 (1H, d, J = 14.0 Hz), 5.50 (1H, d, J = 14.0 Hz), 7 .70 (2H, d, J = 9.0 Hz), 8.10 (2H, d, J = 7.0 Hz), 8.25 (2H, d, J = 9.0 Hz) and 8.90 (2H, d, J = 7.0 Hz).

IR (KBr) Ymax: 3400, 1770, 1690 and 1640 cm-1.

Analysis calculated for C 23 H 24 N 3 O 6 SJ, H 2 O: C, 44.39; H, 4.22; N, 6.82; S, 5.20.

Found: C, 44.66; H, 4.01; N, 6.84; S, 5.64.

B.

15 I® OH B -.

I 'W 3 Pd / c)

Jr -, - ^ o2PNB

20 2? H? J-V9 * '* ·, - ^ N-CH3 g N --CO2® 25 i

To a solution of 779 mg (1.27 mmol) of compound 3 in tetrahydrofuran-water-diethyl ether (80 mL to 80 mL - 100 mL) was added 140 mg (1.4 mmol) of potassium bicarbonate and 125 mg (0 mL) of .7 mmol) dibasic potassium phosphate. 700 mg of a catalyst containing 10% palladium on charcoal were then added and the mixture was hydrogenated at 276 kPa for 45 minutes on a Parr shaker. The mixture was then filtered and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with diethyl ether (150 mL) and then lyophilized to give a brown powder. This crude material was purified on a Cg BONDAPAK reverse phase column (30 g) (Waters Associates), eluting with water at 55 kPa. Each fraction (20 ml) was screened by high pressure liquid chromatography, and fractions with ultraviolet absorption at λ max = 300 nm were pooled and lyophilized to give 135 mg (32% yield) of the title compound 4 as a slightly yellow solid.

NMR (D 2 O) 1.25 (3H, d, J = 6.0 Hz), 2.7-3.2 (2H, m), 3.40 (1H, q, J = 9.0 and 2.5 Hz), 3.9-4.2 (2H, m), 4.40 (3H, s), 4.72 (2H, s), 8.10 (2H, d, J = 6.0 Hz), 7.72 (2H, d, J = 6.0 Hz).

IR (KBr) / max: 3400, 1755, 1640 and 1590 cm-1.

UV Λ. max (H 2 O): 296 nm (ε = 7782), 258 nm (ε = 6913).

Example 2 20 (R) \ | \ = - /} -N-e 25 0 CO2 38 7 8 0 9 4 OH N, (ft) _v __ΝΗ ^ 0 Ο ι

JH

'' C02PNB

2 OH _ J · * -. - S CH - CH \ π I 2 2 w

o * '' C02PNB

3 PNB = p-nitrobenzyl 39,78094

A suspension of 1.1 g (2.93 mmol) of the diazo compound 1 in 30 ml of dry benzene was purged with nitrogen at room temperature for 5 minutes. The suspension was treated with 25 mg of rhodium acetate dimer and the mixture was heated at reflux for 45 minutes. The warm solution was diluted with ethyl acetate (25 ml), filtered to remove the catalyst and evaporated to dryness to give the keto compound 2 as a white solid. This was dissolved in dry acetonitrile (10 ml) and cooled to -10 ° C. To this solution was added, under nitrogen protection, diisopropylethylamine (417 mg, 3.2 mmol) followed by 810 mg (3.0 mmol) of diphenyl chlorophosphate and the reaction mixture was stirred for 20 minutes at -10 ° C. The reaction mixture was then treated with diisopropylethylamine (420 mg, 3.2 mmol) and 2- (4-pyridyl) ethanethiol (560 mL; 4.03 mmol) in 2 mL of acetonitrile ζϊ. Org. Chem. 26:82 (1961) Ludwig Bauer and Libero A. Gardella Jr .J. The reaction mixture was stirred at -5 ° C to -10 ° C for 1 hour, then diluted with methylene chloride (100 mL) and washed sequentially with brine (1: 1), 4% H 2 O with 5% NaHCO 3, IVO and brine. The organic phase was dried (MgSO 4) and evaporated to dryness to give a white solid. This solid was washed with diethyl ether-hexane (1: 1) and dried under high vacuum to give 901 mg (63.9%) of compound 3.

IR (KBr) 1790, 1690 cm -1.

NMR (CDCl 3 / DMSO) δ: 1.20 (3H, d, J = 3.0 Hz, CH 3), 2.8-3.2 (7H, m) 3.9-4.4 (2H, m ), 5.1 (1H, d), 5.4 (2H, q), 7.3 (2H, d), 8.5 (2H, q), 7.76 (2H, d), 8.3 (2H, q).

78094 40 B.

0 ^ ~ N-S: o2pnb 3 10 OH _ e i-n -

15 "COjPNB

4

A suspension of carbapenem 3 (890 mg, 1.85 mmol) and 7 mL of iodomethane in 200 mL of dry acetone and 12 mL of methylene chloride was stirred at 25 ° C for 24 h. The reaction mixture became a clear solution after 18 hours. The solvent was removed in vacuo, then the residue was washed with diethyl ether to give 920 mg (1.48 mmol) (79.8%) of compound 3 as a foamy solid.

IR (KBr) 1765, 1690 cm -1.

NMR (DMSO) δ 1.3 (3H, d, J = 3.0 Hz), 3.1-3.7 (7H, m), 4.1 (3H, m), 4.3 (3H, s ), 5.38 (2H, q, J = 7.0 Hz), 8.1 (2H, d, J = 3.0 Hz), 8.9 (2H, d, J = 3.0 Hz) , 7.6 (2H, d, J = 4.0 Hz), 8.2 (2H, d, J = 4.0 Hz).

78094 41 j- | - (H) y

c / 7 N CO-PNB

5 0 2 4

OH

Π I w f ~ '"^« 2 * 15 i

Carbapenem 4 (920 mg, 1.47 mmol) was treated, dissolved in 90 mL of tetrahydrofuran, 90 with no diethyl ether and 90 with no water, with 265 mg of 20 (1.51 mmol) of dibasic potassium phosphate, with 190 mg of (1.9 mmol). potassium bicarbonate and 800 mg of 10% palladium-on-carbon catalyst. Hydrogenated at 310 kPn pressure for one hour. The catalyst was filtered off through Celite and the filtrate was washed with di-ethyl ether (3 x 25 mL). The aqueous layer was lyophilized to give a brownish substance, which was then purified * twice by chromatography on a 12 g C18 column (H 2 O) to give 55 mg of compound 5.

IR (KBr) 1750, 1640 cm -1.

NMR (D 2 O) δ 1.30 (3H, d, J = 3.0 Hz =, 3.0-3.5 (7H, m), 4.3 (3H, s), 4.0-4 Δ (3H, m), 7.90 (2H, d), 8.70 (2H, d).

* C18 BONDAPAK Reverse Phase Column (Waters Associates) 35 42 78094

OH

λΤΓν -O® 5 ° “f

Example 3 Preparation of 3- (N-methylpyridin-3-ylmethanethio) (R) -β-hydroxyethyl β-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate___

.O

15 1) N—

OH

TT \ o - C1 PC ° 03 2 - ° t. ^ O7

. I

"Λρ> · -ο

C02pNB

30β-Nitrobenzyl-3- (pyridin-3-yl-methanethio) -6e- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [2,7] hept-2-ene-2- carboxylate_ 43 780 9 4 To a cooled solution (0 ° C) of 925 mg (2.66 mmol) of the keto intermediate 5 in 14 mL of acetonitrile was added a solution of 377 mg (2.9 mmol) of diisopropylethylamine 1 acetonitrile 5 followed by 786 mg (2.9 mmol) of diphenyl chlorophosphonate in 1 ml of acetonitrile under a nitrogen atmosphere was stirred for 15 minutes at 0 ° C and then a solution of 377 mg ( 2.9 mmol) of 3-mercaptomethyl-pyridine prepared by the method described in Can. J.

Chem., 56, 3068 (1978) in 7 ml of acetonitrile. The reaction solution was stirred for 90 minutes at 0 ° C. The precipitate was collected by filtration and washed with 20 ml of ethyl acetate to give 950 mg (yield 60%) of the title compound as white crystals.

NMR (DMSO-d 6) δ f: 1.30 (3H, d, J = 6.0 Hz), 3.4-4.2 (5H, m), 4.25 (2H, s), 5.1 (1H, d, J = 4.5 Hz), 5.40 (2H, ABq, J = 14.4 Hz), 7.2-8.5 (8H, m).

IR (KBr) / * max. 3500, 1775 and 1580 cm-1.

Analysis calculated for C 22 H 21 J 3 O 6 S 1: C, 58.01; H, 4.65; N, 9.23; S, 7.04; Found: C, 57.19; H, 5.19; N, 8.76; S, 7.08.

44 78094

OH

A — r -— \ fAl ch i. 7

19 C ° 2PNB J

OH

> 'r-rv -O®

o I

CO, p N * B

20 15 Pd / C, H2

OH V

(AIO ^ CH3 20 ° ‘I Θ

COO

21 3- (N-Methylpyridin-3-yl-methanethio) (R) -25-hydroxyethyl] -7'-oxo-1-azabicyclo (3,2,0 </ hept-2-ene-2-carboxylate)

To a solution of 730 mg (1.56 mmol) of compound 19 in 120 ml of acetone was added 5 ml of methyl iodide, and the reaction mixture was stirred for 18 hours at room temperature. The precipitate was collected by filtration and washed with acetone (10 ml) to give 940 mg (100% yield) of quaternized pyridine as a slightly yellow powder.

45 7 8 0 9 4 NMR (DMSO-d 6) δ: 1.25 (3H, d, J = 5.8 Hz), 3.6-4.3 (5H, m), 4.20 (3H, s ), 4.25 (2H, s), 5.25 (1H, d, J = 7.2 Hz), 5.40 (2H, ABq, J = 12.16 Hz), and 7.6-9, 2 (9 H, m).

IR (KBr) λmax: 3300, 1765 and 1690 cm -1.

Analysis calculated for C23H24N3®6 ^ 1J1: C, 46.24; H, 4.05? N, 7.03; S, 5.37. Found: C, 45.82; H, 4.11; N, 6.87; S, 6.10.

To a solution of 933 mg (1.6 mmol) of compound 20 in 90 mL of tetrahydrofuran and 90 mL of ether-10 were added 200 mg of KHCO 3 and 349 mg of 1 H HPO in 90 mL of water and its then 1.0 g of palladium charcoal. The mixture was hydrogenated at 310 kPa on a Parr shaker for 45 minutes. The soes were filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow solid which was purified on a Cg BONDAPAK (Waters Associates) reverse phase column (8 g) eluting with 5% acetonitrile in water at 8 psi: n painees-20 sa. Each 15 ml fraction was analyzed using high performance liquid chromatography and the fractions with ultraviolet absorption at λ max. 300 nm, collected and lyophilized to give 230 mg (43% yield) of the title product as slightly yellow crystals, m.p. 130 ° C 25 (dec.).

NMR (D 2 O) δ. 1.25 (3H, d, J = 7.0 Hz), 3.12 (2H, dd, J = 7.9 Hz, 1.6 Hz), 3.42 (1H, q, J = 7.2) Hz, 1.6 Hz), 3.9-4.6 (3H, m), 4.25 (2H, s), 4.42 (3H, s) and 8.0-9.0 (4H, m). ).

IR (KBr) ν max: 3400, 1750 and 1580 cm-1.

UV Xmax. (H 2 O): 298 nm (ε = 8058).

Analysis calculated for c 16 H 18 N 2 O 4 S 1 * 2H 2 O! C, 51.87; H, 5.44; N, 7.56; Found: C, 51.95; H, 5.66; N, 7.56.

78094 46

OH

TQ - 9.

5 0 '® CH® CO 2 U 3

Example 4 Preparation of 3- (N-methylpyridin-2-yl-methanethio) (R) -hydroxyethyl-7- (oxo-1-azabicyclo [2.2.1] hept-2-ene-2-carboxylate) N— ° HO "C02pNB HS 'S j 25 /

OH

COIN'S

22 47 7 8 0 9 4 P-Nitrobenzyl-3- (pyridin-2-yl-methanethio) - (R) -hydroxyethyl-7-oxo-1-azabicyclo- (3,2,0) hept-2-ene- 2-Carboxylate To a cooled solution (0 ° C) of 925 mg of 5 (2.65 mmol) of keto intermediate 5 in 14 mL of acetonitrile was added a solution of 377 mg (2.92 mmol) of di- isopropylethylamine in 1 ml of acetonitrile followed by 786 mg (2.90 mmol) of diphenyl chlorophosphate in 1 ml of acetonitrile 10. The resulting solution was stirred for 15 minutes at 0 ° C and then a solution of 377 mg was added. (2.92 mmol) diisopropylethylamine in 1 ml acetonitrile followed by 350 mg (3.0 mmol) 2-mercaptomethylpyridine prepared by the method described in Can. J. Chem., 56, 3068 (1978) in 1 ml of acetonitrile. The reaction solution was stirred for 2 hours at -10 ° C. The precipitate was collected by filtration and washed with 20 ml of methylene chloride to give 650 mg (yield 54%) of the title product as a yellow powder.

NMR (DMSO-d 6) δ: 1.26 (3H, d, J = 7.0 Hz), 2.7-3.5 (4H, m), 3.9-4.3 (2H, m), 4.2 (2H, s), 5.42 (2H, ABq, J = 14.4 Hz) and 7.2-8.8 (8H, m).

IR (KBr) λmax: 3400, 1775 and 1690 cm-1.

Analysis calculated for 022 ^ 2 ^ 3 ^ 6 ^ 1 ^ C, 58.01; H, 4.65; N, 9.23; S, 7.04; Found: C, 57.56; H, 4.92; N, 8.94; S, 7.03.

48 78094

^ JT

Ί r \ .s ^ L · ϋ “v

Nasetone and CO_pNB / 22 2 / OH / xrry, X) 0 CO 2 pNB CH 3 I © 23 15

3- (N-methylpyridin-2-ylmethanethio) -6H-OH H2 (R) - Hydroxyethyl / -7-oxo-1-azabicyclo (3,2,0) -hept-2-ene-2-carboxylate ______

To a solution of 650 mg (1.39 mmol) of compound 22 in 100 mL of acetone was added 4 mL of methyl iodide. The reaction mixture was stirred for 3 days at room temperature. The precipitate was collected by filtration and washed with acetone (10 mL) to give 500 mg (60% yield) of quaternized pyridine 23 as a slightly yellow solid.

NMR (DMSO-d 6): 1.26 (3H, d, J = 7.0 Hz), 2.9-4.2 δ (2H, m), 4.4 (3H, s), 4.78 ( 2H, s), 5.2 (1H, d, J = 3.9

Hz), 5.50 (2H, ABq, J = 14 Hz) and 7.8-9.4 (8H, m).

IR (KBr) ν max: 3400, 1765 and 1690 cm -1.

Analysis calculated for C 23 H 24 N 3 O 6 • 1: 1: C, 46.24; H, 4.05; N, 7.03; S, 5.37; Found: C, 45.62; H, 4.27; N, 6.80; S, 5.30.

To a solution of 1.0 g (1.167 mmol) of compound 23 in 90 mL of tetrahydrofuran and 90 mL of ether was added 215 mg (2.15 mmol) of KHCO 3 and 374 mg (2.1 mmol) of K 2 HPO 2 in 90 mL: in water followed by 1.0 g of 10% palladium on carbon. The mixture was hydrogenated at 310 kPa on a Parr shaker for 45 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 200 mL) and lyophilized to give a yellow solid which was purified on a Cg BONDAPAK (Waters Associates) reverse phase column (10 g) eluting with 5% aqueous acetonitrile in 8 psi. at pressure. Each of the 15 ml fractions was analyzed using high performance liquid chromatography and the fractions with ultraviolet absorption at max. 300 nm were pooled and lyophilized to give 390 mg (44% yield) of the title product. Recrystallization of this material from water-acetone-ethanol gave fine needles, m.p. 194-196 ° C (dec.).

NMR (D 2 O) δ: 1.30 (3H, d, J = 6.2 Hz), 3.2 (2H, q, J = 9.0 Hz), 3.6 Hz), 3.46 (1H , q, J = 6.0 Hz, 2.7 Hz), 4.1-4.6 (3H, m), 4.60 (3H, s), and 7.9-8.9 (4H, m ).

IR (KBr) fmax: 3400, 1755 and 1590 cm-1.

UV A.max. (H2 =): 292 nm (ε = 8092).

35 50 7 8 0 9 4

Analysis calculated for C 28 H 28 N 2 O 4 S 1 · C: 51.87; H, 5.44; N, 7.56; Found: C, 51.37; H, 5.69; N, 7.37.

OH

ο I CH-

10 LQ

®2

Example 5 3- (N-Methylpyridin-2-yl-ethathio) -6- [4- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3,2,0) -15 hept-2- Preparation of ene-2-carboxylate (* s>, OLPNB LM -: 25/5 54 / OH /

J

C02PMB

53 35 51 78094 p-Nitrobenzyl-3- (pyridin-2-yl-ethanethio) -6H- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo- (3,2,0 / hept-2 -ene-2-carboxylate To a cooled solution of 1.78 g (5.0 to 5 mmol) of the keto intermediate 5 in 25 mL of acetonitrile was added 710 mg (5.5 mmol) of diisopropylethylamine in 1 mL under a nitrogen atmosphere. acetonitrile followed by 1.4 g (5.0 mmol) of diphenyl chlorophosphate in 1 ml of acetonitrile The resulting solution was stirred for 20 minutes at 0 ° C and then a solution of 710 mg (5.5 mmol) of diphenyl chlorophosphate was added. ) diisopropylethylamine in 1 ml of acetonitrile followed by a solution of 850 mg (6.1 mmol) of thiol 54 prepared by the method described in J. Org.

15 Chem., 26, 82 (1961) in 2 ml of acetonitrile. The reaction mixture was stirred for 60 minutes at 0 ° C. The precipitate was collected by filtration and washed with methylene chloride (20 mL) to give 1.3 g (57%) of the title product as a yellow solid.

NMR (CDCl 3) δ 1.25 (3H, d, J = 6.5 Hz), 2.6-3.4 (7H, m), 4.2-4.6 (2H, m), δ, 30 and 5.65 (1H, ABq, J = 14 Hz each) and 7.2-8.5 (8H, m).

IR (KBr) ν max: 3400, 1780 and 1680 cm -1.

52 78094 r ch3 'C02PNB / 53 0H /> · w-v n A /, _ C0, PNB CH- 1 15 Pd / C 2 3

Ho 2 55) / ··· »

OH

20 7 '._. ^ X

3- (N-methyl-pyridin-2-yl-ethanethio) (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept-2- ene-2-karboksylaatti_

To a suspended solution of 800 mg (1.7 mmol) of compound 53 in 50 mL of acetone was added 5 mL of methyl iodide. The reaction mixture was stirred for 48 hours at room temperature. The precipitate was collected by filtration and washed with acetonitrile (15 mL) to give 810 mg (76% yield) of quaternized pyridine 55 as a slightly yellow powder.

NMR (DMSO-d 6) δ f: 1.20 (3H, d, J = 5.6 Hz), 3.2-4.3 δ (9H, m), 4.20 (3H, s), δ, 26 and 5.55 (1H, ABq, J = 15 Hz each) and 7.8-9.2 (8H, m).

IR (KBr / * max: 3400, 1770 and 1690 cm-1

To a solution of 790 mg (1.27 mmol) of compound 55 in 100 mL of tetrahydrofuran and 100 mL of ether-10 blades was added 100 mL of pH = 7 buffer, followed by 1.0 g of 10% palladium on charcoal. The mixture was hydrogenated at 276 kPa on a Parr shaker for 40 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow powder which was purified on a Cg BONDAPAK (Waters Associates) column (30 g) eluting with 10% acetonitrile in water at 8 psi.

Each 15 ml fraction was analyzed by high performance liquid chromatography and the fractions with ultraviolet absorption at Amaks were pooled. 300 nm and lyophilized to give 65 mg (15% yield) of the title product as a yellow powder.

NMR (D 2 O) δ s 1.25 (3H, d, J = 6.2 Hz), 3.1-3.6 (7H, m), 4.0-4.3 (2H, m), 4 .32 (3H, s) and 7.8-8.9 (4H, m).

IR (KBr) fmax: 3400, 1750 and 1590 cm -1.

UV λmax (H 2 O): 300 nm (ε = 8108).

30 54 7 8 0 9 4

OH

, λρ ^ · ~ ο ~ CO ©

Example 6 3- (1-Propylpyridin-4-yl-methanethio) -6β- [1- (R) -10-hydroxyethyl] -7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2 preparation of carboxylate

OH 0H

15 Xx. Xs s ___. __,

P> -fQ-O

Z- C0, pNB

COjPNB 2

20 J5 *> I

p-Nitrobenzyl-3- (pyridin-4-yl-methanethio) -N- (6H-1- (R) -hydroxyethyl) -7-oxo-1-azabicyclo-0.2,0-hept-2-ene- 2-Carboxylate A solution of 673 mg (1.86 mmol) of p-nitrobenzyl-6H-1- (R) -hydroxyethyl] -3,7-dioxo-1-aza-bicyclo [3.2.07] hept. 2-ene-2-carboxylate (5) in 10 ml of acetonitrile, cooled to -10 ° C under a nitrogen atmosphere. A solution of 245 mg (1.90 mmol) of diisopropylethylamine in 1 mL of acetonitrile was added, followed by dropwise addition of 510 mg (1.90 mmol) of diphenyl chlorophosphate in 1 mL of acetonitrile over 2 minutes. The resulting solution was stirred at -10 ° C for 15 minutes to give p-nitrobenzyl-3- (diphenylphosphoryloxy) -6? - [1- (R) -hydroxyethyl] -7-oxo. -1-azabicyclo (3.2.0 (hept-2-ene-2-carboxylate) To this solution was added a solution of 245 mg (1.90 moles) of diisopropylethylamine in 5 ml. acetonitrile followed by a solution of 270 mg (2.16 mmol) of 4-mercaptomethylpyridine in 0.5 mL of acetonitrile The reaction mixture was stirred at -10 ° C for 60 minutes and the resulting white precipitate was collected by filtration and washed with 5 mL. : Ice-10 cold acetonitrile to give 660 mg (yield 76%) of compound 51 as white crystals, mp 145 ° C.

NMR (DMSO-d 6) δ: 1.20 (3H, d, J = 6.0 Hz), 3.2-3.4 (3H, m), 3.7-4.1 (2H, m), 4.25 (2H, s), 5.05 (1H, d, J = 4.0 Hz), 5.35 (1H, d, J = 14.0 Hz), 5.48 (1H, d, 15 J = 14.0 Hz), 7.40 (2H, d, J = 5.5 Hz), 7.70 (2H, d, J = 8.5)

Hz), 8.23 (2H, d, J = 8.5 Hz) and 8.58 (2H, d, J = 5.5 Hz).

IR (KBr) fmax: 3400, 1790 and 1695 cm -1.

Analysis calculated for C 22 H 21 N 3 O 6 S: C, 58.01; H, 4.56; N, 9.23; S, 7.04; Found: C, 57.74; H, 4.56; N, 9.58; S, 7.21.

OH

'25 J - \ - / / O | Na I / CO 2 pNB / 51 0H / 0 CO 2 pNB I '52 35 56 7 δ O 9 4 3- (1-Allylpyridin-4-ylmethanethio) -6? - (?) - (R) -hydroxyethyl,? - 7- oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate_

To a solution of 900 mg (2.13 mmol) of compound 51 in 150 mL of acetone was added 2 mL of allyl bromide and 380 mg of sodium iodide. The mixture was stirred for 48 hours at room temperature and the solvent was evaporated off in vacuo to give a yellow solid. This material was suspended in 120 of any acetonitrile, filtered and evaporated to dryness in vacuo to give 1.0 g (87% yield) of the title product as a yellow solid.

NMR (CD 3 CN) 1.20 (3H, d, J = 6.2 Hz), 3.0-3.4 (4H, m), 4.0-4.4 (4H, m), 5.1- 5.6 (4H, m) and 7.4-79 (8H, m).

IR (KBr) fmax: 3400, 1770 and 1690 cm -1.

Analysis calculated for C 25 H 26 N 3 O 6 S 1 J 1: C, 48.16? H, 4.21; N, 6.74; S, 5.15; Found: C, 48.55? H, 4.46; N, 6.69? S, 5.15.

20

OH

CO 2 pNB 1 "/ β-3'-1" s 53,357,78094 3- (1-propylpyridin-4-yl-methanethyl) -6 * 4 - [(1-) hydroxy-ethyl] -7- oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate_

To a solution of 1.27 g (2.15 nunols) of compound 52 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer followed by 1.0 g of 10% palladium. -puuhiiltä. The mixture was hydrogenated at 276 kPa on a Parr shaker for 40 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow powder which was purified on a Cg BONDAPAK (Waters Associates) column (40 g) eluting with 10% acetonitrile in water at 8 psi. . Each 15 ml fraction was analyzed and the fractions with ultraviolet absorption at Xmax. 300 nm, combined and lyophilized to give 48 mg (6% yield) of the title product as a yellow powder.

NMR (D 2 O) δ: 0.95 (3H, t, J-7.5 Hz), 1.25 (3H, d, 7.0 Hz), 2.05 (2H, sextet, J = 7.5 Hz), 3.10 (2H, dd, J = 10 Hz, 2.5 Hz), 3.35 (1H, dd, J = 6.5 Hz, 2.5 Hz), 4.0-4.8 (6H, m), 7.1 (2H, d, J = 6.0 Hz) and 7.80 (2H, d, J = 6.0 Hz).

IR (KBr) ν max: 3400, 1750 and 1590 cm -1.

Analysis calculated for C 18 H 22 N 2 O 4 S · 2H 2 O: C, 54.52; H, 6.10; N, 7.07; Found: C, 54.32; H, 6.03; N, 6.99.

58 78094

OH

, CO® «3

Example 7 Preparation of 3- (N-methyl-3-methylpyridine-2-methanethio) -10 (R) -hydroxyethyl-7- (oxo-1-azabicyclo-D, 2,0] hept-2-ene-2-carboxylate C &

2) NaOH

HCl 36 '37 20 3-Methyl-2-mercaptomethyl-pyridine

A solution of 2.45 g (17.0 mmol) of the chlorine compound 36 and 1.37 g (18.0 mmol) of thiourea in 60 ml of absolute ethanol was heated at reflux for 5 hours. Evaporation of the ethanol followed by addition of ether gave 3.08 g (72% yield) of the isothiouronium salt which was dissolved in 10 ml of water containing 1.44 g (26 mmol) of sodium hydroxide. The solution was then heated at 100 ° C for 5 minutes under a nitrogen atmosphere. The reaction mixture was cooled to 5 ° C, the pH was adjusted to 6.4 by the addition of acetic acid and extracted with ether (4 x 50 mL). The combined ether extracts were washed with 5% aqueous sodium bicarbonate and brine. Evaporation of the solvent from the dried solution (MgSO 4) gave 1.4 g (83% yield) of thiol 37 as a yellow to 35 oil which was used in the next step without further purification.

NMR (CDCl 3 δ: 2.20 OH, s), 2.5-2.7 (1H, broad s), 3.8 (2H, t, J = 6.5 Hz) and 6.9-8, 2 (3 H, m).

OH ^ X "ΓΤΛ = 0 C1P (0W *.

10 J

C ° 2pNB / s "'" V /

15 OH

X ^ "TVs ^ f \ \ = /

20 CO2pNB

38 25 p-Nitrobenzyl-3- [3-methylpyridin-2-yl-methanethiol] -6H- (R) -hydroxyethyl-4- (7-oxo-1-azabicyclo [3.2.1] hept-2-ene) -2-Carboxylate To a cooled (0 ° C) solution of 1.74 g (5.0 mmol) of Keto Intermediate 5 in 25 mL of aceto-30-nitrile was added 960 mg (5.8 mmol) of di- isopropylethylamine in 2 ml of acetonitrile followed by 1.4 g (5.8 mmol) of diphenyl chlorophosphate in 2 ml of acetonitrile The resulting solution was stirred for 20 minutes at 0 ° C and then a solution of 760 mg ( 5.8 mmol) of di-60 7 8 0 9 4 isopropylethylamine in 2 ml of acetonitrile followed by 810 mg of mercaptomethylpyridine 37 in 3 ml of acetonitrile The reaction mixture was stirred for 2 hours at 0 [deg.] C. The precipitate was collected by filtration and washed with acetone-5. to give 1.56 g (66% yield) of the title product as a white solid, mp 145 ° C.

NMR (DMSO-d 6) δ: 1.23 (3H, d, J = 6.5 Hz), 2.30 (3H, s), 3.1-4.3 (6H, m), 4.35 ( 2H, s), 5.20 and 4.45 (1H each, ABq, J = 15.0 Hz) and 7.3-8.4 (7H, m).

10 IR (KBr) λmax: 3400, 1767 and 1695 cm-1.

Analysis calculated for C 24 H 26 N 3 O 9 • 2F: C, 47.91; H, 4.69; N, 6.98; S, 10.66; Found: C, 47.72; H, 4.34; N, 6.72; S, 11.22.

15 OH

\ M

y-j ^ v _fso3ch3 20 0 CO2pNB / OH / ”A c% 25 \ —n - = - / ° CO2pNB CH37 37

I Pd / C H 0SO3F

ψ H2

OH

30 μl CO 2 ° CH 3 40 35 61 78094 3- (N-methyl-3-methylpyridin-2-ylmethanethio) -64- (1-) - (hydroxy) ethyl] -7-oxo-1-azabicyclo- (3, 2.0) hept-2-ene-2-karboksylftatti_

To a solution of 680 mg (1.45 mmol) of compound 38 in 120 mL of methylene chloride was added 270 mg (2.33 mmol) of methyl fluorosulfonate. The reaction mixture was stirred for 3 hours at room temperature. The precipitate was collected by filtration and washed with methylene chloride (5 mL) to give 840 mg (99% yield) of quaternized pyridine 39 as white crystals.

NMR (DMSO-d 6): 1.15 (3H, d, J = 5.8 Hz), 2.62 (3H, s), 3.2-4.4 (5H, m), 4.45 (3H , s), 4.60 and 4.82 (1H, ABq, J = 9.2 Hz each), 5.30 and 5.46 (1H, ABq, J = 12.8 Hz each) and 7.6- 8.9 (7 H, m).

IR (KBr) ν max: 3400, 1750 and 1590 cm-1

Analysis calculated for C 24 H 24 N 3 O 9 S 2 F: C, 49.14; H, 4.47; N, 7.13; S, 11.43; Found: C, 49.56; H, 4.16; N, 7.26; S, 11.03.

To a solution of 810 mg (1.39 mmol) of compound 39 in 100 mL of tetrahydrofuran and 100 mL of ether was added 100 mL of pH 7.0 buffer followed by 750 mg of 10% palladium on charcoal. The mixture was hydrogenated at 241 kPa on a Parr shaker for 60 minutes in a cold room (4-6 ° C). The mixture was filtered through a pad of Celite-25 and the catalyst was washed with ether (2 x 10 mL).

The combined filtrate and washings were extracted with ether (2 x 40 mL) and lyophilized to give a yellow solid which was purified on a Cg BONDAPAK (Waters Associates) column (20 g) eluting with 5% acetonitrile in water at 8 psi.

Each 15 ml fraction was analyzed using high performance liquid chromatography and fractions with ultraviolet absorption at Amaks were pooled. 300 nm, and lyophilized to give 141 mg (30% yield) of the title product 35 as a yellow solid.

62 7 8094 NMR (D 2 O) 1.24 (3H, d, δ = 1.0 Hz), 2.62 (3H, s), 3.2-3.5 (3H, m), 4.2-4 , 4 (2H, m), 4.45 (3H, s), 4.50 and 4.59 (1H, ABq, J = 12.6 Hz each), 7.82 (1H, q, 3 = 1, 0 Hz, 6.5 Hz), 8.35 (1H, d, J = 7.0 Hz) and δ 8.65 (1H, d, J = 6.5 Hz).

IR (KBr) ν max: 3400, 1750 and 1580 cm -1.

UV 7-tnaks. (H 2 O): 296 nm (ε = 8014).

Analysis calculated for C 17 H 20 N 2 O 4 S 1 • H 2 O 2: C, 57.85; H, 5.85; N, 7.94; Found: C, 58.60; H, 5.86; N, 7.87.

OH

15 \

0 (CO ® K

Example 8 3- (2-Methyl-N-methylthiazol-4-yl-methanethio) -N, O-1- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo- (3,2,0) heptene Preparation of -2-carboxylate 15> - £ - <

25 OH

-r ^ \ cip (o0) 2 / = ° ---> »s? -2) / T-s

0 Ξ HS I

30 ° CPNB 2 CH 3 5

X

C02pNB

9,6378094 p-Nitrobenzyl-3 - [(2-methylthiazol-4-yl) methanethiol-6- [1- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [2,2] hept-2-ene -2-carboxylate To a cooled (0 °) solution of 1.4 g of 5 (4.0 moles) of keto intermediate 5 in 12 mL of acetonitrile was added 0.83 mL (4.6 mmol) of di -isopropylethylamine followed by 1.16 g (4.3 mmol) of diphenyl chlorophosphate in 2 ml of acetonitrile and the resulting solution was stirred at 0 ° for 10 minutes to give p-nitrobenzyl-3- (diphenylphosphoryloxy) (R) -hydroxyethyl 7-7-oxo-1-azabicycloC3,2,0-hept-2-ene-2-carboxylate To this solution was added a solution of 0.83 ml (4.6 mmol) of diisopropylethylamine in 2 ml of acetonitrile. 15 liters followed by a solution of 0.62 g (4.2 mmol) of 2-methyl-4-mercaptomethylthiazole prepared by the method described in J. Amer. Chem. Soc., 71, 3570 (1949). In 3 ml of acetonitrile The reaction solution was stirred 40 minutes at 0 ° C. The precipitate was collected and washed with ether (30 mL) to give 943 mg of the title product as a white solid.

NMR (CDCl 3) δ 1.32 (3H, d, J = 7 Hz), 2.68 (3H, s), 3.20 (2H, m), 3.76 (1H, d, J = 5, 5 Hz), 4.16 (2H, s), 4.20 (1H, m), 5.40 (2H, q, J = 14 Hz), 7.06 (1H, s), 7.68 ( 2H, d, J = 8 Hz) and 8.24 (2H, d, J = 8 Hz).

IR (KBr) / max: 3500, 1770 and 1700 cm-1.

64 78094

OH

X ^ ^

I Γ \ _ // S CH OSO F

II A-S'X I -5_J_> 5 θ · ^ ”Ν T XN ^ CH3 ™ 2C12

CO pNB

9

OH

10 ^ rT ^ V-s I

0 ^ ^ 0 CH3 C ° 2 *> NB CH, - FSO_Ö 15 Pd / C, H2 -5

OH V

3- (2-Methyl-N-methyl-thiazol-4-yl-methanethio) - (R) -hydroxyethyl-7- (oxo-1-azabicyclo-2 5,2,2,7-hept-2- enene-2-carboxylate ___, _

To a solution of 525 mg (1.1 mmol) of Compound 9 in 20 mL of methylene chloride was added 0.27 mL (3.3 mmol) of methyl fluorosulfonate. The reaction mixture was stirred for 90 minutes at room temperature. The precipitate was collected by filtration and washed with methylene chloride (50 mL) to give 650 mg (100% yield) of quaternized thiazole 10, which was used in the next step without further purification.

Thus, to a solution of 10 in 100 ml of tetrahydrofuran and 100 ml of ether was added 100 ml of a pH 7.0 buffer solution, followed by 500 mg of 10% palladium on carbon. The mixture was hydrogenated at 241 kPa in a Parr shaker for 45 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow powder which was purified on a Cg BONDAPAK reverse phase column (8 g) Water Associates eluting with 5% acetonitrile in water at 8 psi. Each 15 ml fraction was analyzed using high pressure liquid chromatography and fractions with 10 ultraviolet absorption at Xmax. 300 nm, combined and lyophilized to give 145 mg (48% yield) of the title compound as a pale yellow powder.

NMR (CDCl 3 1.20 (3H, d, J = 7 Hz), 2.92 (3H, 15S), 3.08 (1H, d, J = 3.5 Hz), 3.20 (1H, d , J = 3 Hz), 3.44 (1H, dd, J = 1 Hz, J = 3.5 Hz), 4.00 (3H, 5), 4.20 (3H, m), 4.36 ( 2H, m) and 7.88 (1H, s).

IR (KBr) ν max: 3400, 1750 and 1585 cm-1.

UV Amaks. (H 2 O): 296 nm (ξ = 7500).

Analysis calculated for C 18 H 18 N 2 O 2 S 2 .2H 2 O: C, 46.15; H, 5.64; N, 7.17; S, 16.41. Found: C, 46.50; H, 5.26; N, 7.13; S, 16.20.

Example 9 3- (N, N'-Dimethylimidazol-2-yl-methanethio) -6H-1- (R) -hydroxyethyl-7-oxo-1- Preparation of azabicyclo (3,2,0) hept-2-ene-3-carboxylate 66,78094 CH ° ·}

I s I

A “uH

I ---> II // ^ SH

5 ^ —jj 2) EtOH, Δ HCl HCl

II II

2-Mercaptomethyl-N-methylimidazole To a solution of 10.4 g (58 mmol) of 2-chloromethyl-N-methylimidazole 31 was prepared by the method described in J. Amer. Chem. Soc., 71,383 (1949) in 7,200 ml of acetonitrile, 7.1 g (60 mmol) of N-acetylthiourea were added and the reaction mixture was heated at reflux for 90 minutes. The precipitate was filtered off and washed with acetonitrile (20 ml) to give the isothiouronium salt, which was then dissolved in 120 ml of ethanol and heated at reflux for 18 hours under a nitrogen atmosphere. The reaction mixture was cooled to 20 room temperature, condensed in vacuo to a volume of about 60 ml and the precipitate was removed by filtration. Evaporation of the filtrate to dryness in vacuo gave 2-mercapto-methyl-N-methylimidazole 32 as a yellow oil which was used in the next step without further purification.

NMR (D 2 O) 3.90 (3H, s), 4.10 (2H, s) and 7.25 (2H, S).

67 78094 1) -N—

OH

I o —r "" \ • Λ> ”vs-] MCI /

10 J

32 *

OH

I CH_

C02pNB

33 20 p-Nitrobenzyl-3- [N-methylimidazol-2-yl-methanethio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo 0.2.0-hept-2 -ene-2-carboxylate To a cooled (0 ° C) solution of 7.24 g of the keto intermediate 5 (35.3 mmol) in 35 mL of acetonitrile was added 2.8 g (21.3 mmol) of di- isopropylethylamine in 2 ml of acetonitrile followed by 5.5 g (20.4 mmol) of diphenyl chlorophosphate in 2 ml of acetonitrile. The resulting solution was stirred at 0 ° C for 15 minutes and then a solution of 4.1 g (3.0 mmol) of diisopropylethylamine in 2 mL of acetonitrile was added followed by 4.6 g (31.0 mmol) of thiol 32. Reaction mixture stirred for 60 minutes at 0 ° C. The white precipitate was collected by filtration and washed with methylene chloride (20 ml) to give 6.6 g (71% yield) of the title product as a white solid, m.p. 142 °.

NMR (DMSO-d 6) 1.32 (3H, d, J = 7.0 Hz), 3.2-4.5 δ (5H, m), 3.2 (2H, s), 3.9 (3H , s), 5.50 (2H, ABq, J = 14.0

Hz), 7.65 (2H, d, J = 6.5 Hz), 7.70 (2H, s) and 8.24 (2H, d, J = 6.6 Hz).

IR (KBr) Ymax: 3450, 1770 and 1690 cm-1.

Analysis calculated for C 21 H 20 N 4 O 6 S 1 N 2 O 2/2 10 C, 52.18; H, 4.79; N, 11.59; Found: C, 52.22; H, 4.91; N, 12.16.

OH Me “- ^ 7 CO2pNB /

,, OH / CH

20 “1 1

"TQ-Q

0 I | © 0 CO 2 pNB CH 3 25 li

Pd / C, H 2 OH V CH 2 CO 2 CH 3 35 35 69 78094 3- (N, N'-Dimethylimidazol-2-yl-methanethio) -6H-4- (R) -hydroxyethyl-7--oxo -1-azabicyclo- (3,2,0) hept-2-ene-2-carboxylate_

To a suspended solution of 1.34 g (3.0 mmol-5 L) of compound in 33,270 mL of acetone was added 20 mL of methyl iodide. The reaction mixture was stirred for 4 days at room temperature. The precipitate was collected by filtration and washed with acetone (20 ml) to give 1.70 g (yield 96%) of quaternized imidazole 34 as yellow crystals, m.p. 175-177 ° C.

NMR (DMSO-d 6) 1.10 (3H, d, J = 6.2 Hz), 3.30 (2H, s), 3.2-4.3 (6H, m), 3.95 (6H, s), 5.45 (2H, ABq, J = 14 Hz), 7.65 (2H, d, J = 6.0 Hz).

IR (KBr) δ max: 3400, 1750 and 1600 cm -1.

Analysis calculated for C 21 H 22 N 4 O 6 S 15 C, 43.08; H, 9.60; N, 5.48; Found: C, 43.02; H, 9.02; N, 5.44.

To a solution of 1.30 g (1.86 mmol) of compound 34 in 120 mL of tetrahydrofuran and 120 mL of ether-20 blades was added 120 mL of pH 7.0 buffer followed by 900 mg of 30% palladium on Celite. The mixture was hydrogenated at 276 kPas on a Parr shaker for 40 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 15 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow amorphous powder which was purified on a 0 x 3 BONDAPAK (Waters Associates) column (30 g) eluting with 10% acetonitrile in water at 55 kPa. . Each fraction of 20 ml was analyzed using oral liquid chromatography and fractions with ultraviolet absorption at max. 300 nm, combined and freeze-dried to give 220 mg (35% yield) of the title product as a yellow powder.

35 NMR (D 2 O) δ: 1.12 (3H, d, J = 7.0 Hz), 3.08 (1H, dd, J = 13.0 Hz, 6.4 Hz), 3.15 (1H, dd, J = 13.0 Hz, 6.4 Hz), 70 78094 3.45 (1H, dd, J = 3.2 Hz, 4.5 Hz), 3.85 (6H, s), 4.1 -4.3 (2H, m), 4.40 (1H, d, J = 13.5 Hz), 4.52 (1H, d, J = 13.5 Hz) and 7.40 (2H, s) .

IR (KBr) λmax: 3500, 1750 and 1590 cm-1.

5 UV max. (H 2 O): 296 nm (ε = 8.411).

Analysis calculated for C 18 H 18 N 2 O 2 S 2 O 2 S 2 O 2: C, 51.68; H, 5.67; N, 12.06; S, 9.50; Found: C, 49.93; H, 5.94; N, 11.46; S, 9.03.

10 oh f CH3V® / “s 15 CO®

Example 10 3- (2,3,4-Trimethyl-thiazol-6-yl-methanethio) - (R) -hydroxyethyl] -7-oxo-1-azabicyclo-20,2,3hept-2-ene-2-carboxylate manufacture. ! s -yi-.

"Ύ K · -A rH

CH 3 H 3 HCl 46 47 30 2,4-Dimethyl-5-mercaptomethyl-thiazole To a solution of 4.8 g (26.0 mmol) of the chlorine compound 46 was prepared by the method described in J. Amer. Chem. Soc., 104, 4461 (1982) ^ 78094 in 50 ml of absolute ethanol, 2.4 g (30 ramols) of thiourea were added. The reaction mixture was heated at reflux for 18 hours. The precipitate was collected by filtration and washed with ether (20 ml) to give the isothiouron-5 salt, which was dissolved in 22 ml of 1-norm. sodium hydroxide and heated for 4 minutes at 100 ° C under a nitrogen atmosphere. The reaction mixture was then cooled to room temperature, the pH was adjusted to 7.0 1-norm. hydrochloric acid and extracted with ether (3 x 50 ml). The combined ether phases were washed with water, brine and dried over MgSO 4.

Evaporation of the dried solution to dryness gave 780 mg (49% yield) of thiol 47 as a colorless oil, which was used in the next step without further purification.

NMR (DC13) δ: 2.05 (3H, s), 2.35 (3H, s) and 3.60 (2H, d, J = 6.5 Hz).

1) N - ^ 20 OH ^ ^

Arrv.

CO * pNB 7 CHS / - 1 30 OH * 'nrVs ^ T ^ 35 CO 2 pNB ^ 3 48 72 7 8 0 9 4 p-Nitrobenzyl-3- [2,4-dimethyl-thiazol-5-yl-methanethiol7- 6e6r / f- (R) -hydroxyethyl7-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate__ To a cooled (0 ° C) solution of 1.4 g of 5 (4 .0 mmol) of the keto intermediate 5 in 25 ml of acetonitrile, 610 mg (4.7 mmol) of diisopropylethylamine in 1 ml of acetonitrile were added under a nitrogen atmosphere, followed by 1.15 g (4.3 mmol) of diphenyl chlorophosphate in 1 ml of acetonitrile. The resulting solution 10 was stirred for 20 minutes at 0 ° C, and then a solution of 610 mg (4.7 mmol) of diisopropylethylamine in 1 ml of acetonitrile was added, followed by 750 mg (4.7 mmol) of thiol. 47 in 2 ml of acetonitrile. The reaction mixture was stirred for 3 hours at 0 ° C. Precipitate 15 was collected by filtration and washed with methylene chloride (20 mL) to give 1.14 g (61% yield) of the title product as a white solid.

NMR (DMSO-d 6) δ: 1.25 (3H, d, J = 6.4 Hz), 2.30 (3H, s), 2.65 (3H, s), 3.1-3.4 ( 3H, m), 4.10 (1H, broad 20 s), 4.0-4.5 (3H, m), 5.25 and 5.50 (1H each, ABq, J = 4 Hz), 7, 68 (2H, d, J = 8.5 Hz) and 8.25 (2H, d, J = 8.5 Hz).

IR (KBr) Vmax: 3500, 1770 and 1690 cm -1.

Analysis calculated for C 22 H 23 N 3 O 6 S 2: 25 C, 53.73; H, 4.71; N, 8.57; S, 13.44; Found: C, 53.97; H, 4.74; N, 8.58; S, 13.10.

78094

OH

JL

rv s ^ svch3 s C02pNB CH \ 48 \ OH \ l — r ^ \ / vc "5 J — P /

o I f CH

CO 2 fNB CH 3 O

SO, F

49 3 15 Pd / C, H2 OH v A / S ^ CH3 Γ /> - s Ά | I®

2o chx ^ f VNX

CO 2 CH 3 50 3- (2,3,4-Trimethyl-thiazol-5-yl-methanethio) -256U- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.2] hept- 2-ene-2-karboksylaatti_

To a solution of 1.97 g (4.0 mmol) of compound 48 in 180 mL of methylene chloride was added a solution of 0.98 mL (13 mmol) of methyl fluorosulfonate in 2 mL of methylene chloride. The reaction mixture was stirred for 70 minutes at room temperature. The reaction mixture was poured into a solution of ether (400 ml) and n-pentane (100 ml). The precipitate was collected by filtration and washed with ether (20 ml) to give 1.6 g of 35 (yield 65.5%) of quaternized thiazole 49 as a white amorphous powder.

74 7 8 0 9 4 NMR (DMSO-d 6) 1.25 (3H, s, J = 6.5 Hz), 2.45 (3H, s), 2.80 (3H, s), 3.2- 4.5 (6H, m), 3.90 (3H, s), 5.30 (2H, broad s), 7.60 and 8.2 (1H each, d, J = 8.5 Hz).

IR (KBr) max 3400, 1770 and 1690 cm-1.

5 Analysis calculated for the compound ^ -ji ^ gNgOgSgF.1 / 2 1 ^ 0; C, 45.09; H, 4.44; N, 6.86;

Found: C, 44.50; H, 4.38; N, 6.58.

To a solution of 1.0 g (1.72 iranol) of compound 49 in 100 mL of tetrahydrofuran and 100 mL of ether-10 was added 100 mL of pH 7.0 buffer followed by 1.0 g of 10% palladium. -puuhiiltä. The mixture was hydrogenated at 276 kPa on a Parr shaker for 40 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (3 x 100 mL) and lyophilized to give a yellow powder which was purified on a Cg BONDAPAK (Waters Associates) column (40 g) eluting with 10% acetonitrile in water at 8 psi.

Each 15 ml fraction was analyzed by high performance liquid chromatography and the fractions with ultraviolet absorption at Amaks. 300 nm, combined and lyophilized to give 315 mg (50% yield) of the title product as a yellow solid.

NMR (D 2 O) δ: 1.25 (3H, d, J = 7.0 Hz), 2.25 (3H, s), 2.90 (3H, s), 3.0-3.30 (3H , m), 3.90 (3H, s) and 4.1-4.4 (4H, m).

IR (KBr) δ max .: 3400, 1750 and 1580 cm -1 UV λmax: (H 2 O): 297 nm (ε = 8994).

Analysis calculated for C 18 H 18 N 2 O 2 S 2 S 2 O 2: C, 48.25; H, 6.09; N, 7.79;

Found: C, 47.96; H, 5.83; N, 7.89.

75 7 8 0 9 4 Γ ^ χ py — o ° I Θ © ^ ° 2 CH3

Example 11 Preparation of 3- [2- (N-methylthiazolium) methylthio] -6% {- (R) -hydroxyethyl.--7-oxo-1-azabicyclo [3.2.1] hept-2-ene-2-carboxylate_

O ^ 0H soq2 fsyp \ a vU

---- H Cl ---- HC1 20, 2 Ο'Ύ '· -► Cr ^ · m 2HCl.

- 30 3 4 2-mercaptomethyl-thiazole

To a solution of thionyl chloride (3.81 mL, 0.052 mol) in chloroform (30 mL) at room temperature was added 3.60 g (0.026 mol) of hydroxymethylthiazole 1 and then heated at 50 ° for 2 hours. . The chloroform was evaporated in vacuo to leave a brown solid which was dissolved in 30 ml of absolute ethanol. Then 2.04 g (0.026 mol) of thiourea was added. The mixture was then heated to reflux for 18 hours. The precipitate was collected by filtration, washed with ethanol and ether to give 3.4 g (yield 55%) of isothiouronium salt 10 3. Isothiouronium salt 3 was dissolved in 30 ml of water and rinsed for 11-20 minutes. 1.10 g (0.027 mol) of sodium hydroxide were added and the mixture was heated at 100 ° for two minutes. The pH of the cooled solution (0 °) was adjusted to 6.0 with acetic acid and then extracted with ethyl acetate (2 x 35 mL). The organic layer was dried (MgSO 4) and evaporated to dryness in vacuo to give 0.75 g (42% yield) of thiol 4 as a yellow oil which was used without further purification; NMR (CDCl 3) δ. 2.1 (1H, t), 4.0 (2H, d, J = 10 Hz), 7.27 (1H, d, J = 3.0 Hz) and 8.85 (1H, d, J = 3.0 Hz).

OH

^ 'nrv · ”25 ci> ρ (0φ) 2 C02PNB 3)

s ^ O

30 5 4 N "" "• m * ° j / 'ΓΤΛ, λ / s / —- f

t0, PNB

6 C

f * 77 7 8 0 9 4 p-Nitrobenzyl-3 - [(2-thiazole) methylthio] -6 ° C- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo- ¢ 3.2.07hept -2-ene-2-carboxylate To a cooled solution (0 °) of 1.4 g of 5 (4.0 mmol) of the keto intermediate 5 in 8 mL of acetonitrile was added 0.79 mL (4, 4 mmol) of diisopropylethylamine followed by 1.17 g (4.4 mmol) of diphenyl chlorophosphate. The resulting solution was stirred at 0 ° for 30 minutes to give p-nit-10-robenzyl-3- (diphenylphosphoryloxy) -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2, To this solution was added a solution of 0.79 ml (4.4 mmol) of diisopropylethylamine in 2 ml of acetonitrile, followed by a solution of 0.72 g of thiol. 4 in 2 mL of acetonitrile The reaction solution was stirred for 60 minutes at 0 ° and then diluted with 50 mL of ethyl acetate and washed with 30 mL of water, 20 mL of 20% aqueous H 3 PO 4 and 30 mL of brine. Evaporation of the solvent from the dried (MgSO 4) solution gave a crystalline solid which was triturated with ether to give 782 mg (42% yield) of the title product 6 as a white crystalline solid, mp 158-160 ° C.

NMR (CDCl 3) 1.32 (3H, d, J = 7.0 Hz), 3.28 (3H, m), 4.20 (2H, m), 4.36 (2H, s), δ, 40 (2H, q), 7.40 (1H, d, J = 4.0 Hz), 7.64 (2H, d, J = 8 Hz), 7.76 (1H, d, J = 4.0 Hz), Hz) and 8.24 (2H, d, J = 8 Hz).

IR (KBr) ν max: 3500, 1770 and 1700 cm -1 Analysis calculated for C 20 H 19 N 3 O 6 • 2: 30 ° C, 52.05; H, 4.15; N, 9.10; S, 13.89; Found: C, 52.35; H, 4.40; N, 8.72; S, 13.90.

78 78094

OH

= = £ CO 2pnb / OH / - J / 'ΓΓ> -Λ

/ π V

'<VNB | H® 3 ie Pd / C, H.

15 OH v 2 FSO3O

J; RQ-s0> - t (@ L3® 3- [2- (N-methylthiazolium) methylthio] -6- [1- (R) -25 hydroxyethyl] -7-oxo-1-azabicyclo [3.2 , 7hept-2-ene-2-carboxylate_

To a solution of 782 mg (1.36 mmol) of compound 6 in 55 mL of methylene chloride was added 0.5 mL of methyl fluorosulfonate and stirred for 90 minutes at 30 room temperature. The precipitate was collected by filtration and washed with methylene chloride (30 ml) and ether (20 ml) to give 630 mg of crude quaternized 79,78094 thiazole 7, which was used in the next step without further purification.

Thus, to a solution of compound 7 in 140 ml of tetrahydrofuran and 120 ml of ether was added 140 ml of a pH 7.0 buffer solution, followed by 650 mg of 10% palladium-on-charcoal. The mixture was hydrogenated at 207 kPa on a Parr shaker for 35 minutes. The mixture was then filtered and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 10 150 mL) and lyophilized to give a yellow powder. The crude yellow powder was purified on a Cg BONDAPAK reverse phase column (7 g) (Waters Associates) eluting with 5% acetonitrile in water at 55 kPa. Each 15 ml fraction was analyzed using high performance liquid chromatography, and fractions with ultraviolet absorption at Xmax. 300 nm, combined and lyophilized to give 23 mg (5% yield) of the title compound as a yellow amorphous solid.

NMR (D 2 O) δ: 1.28 (3H, d, J = 7.0 Hz), 3.12 (2H, d, J = 7.0 Hz), 3.44 (1H, dd, J = 1 .0 Hz and 3.0 Hz), 4.20 (3H, s), 4.24 (2H, m), 4.76 (3H, m), 8.12 (1H, d, J = 4 Hz) and 8.24 (1H, d, J Hz).

IR (KBr) / * max: 3400, 1740 and 1580 cm-1.

25 UV Amaks. (H 2 O) 292 nm (C = 7285).

Γ .. ^ • 'rrv.-C-ry-s / - "f w W * 80 7 8 0 9 4

Example 12 3- {1- (RS) -Methyl-N-methyl-pyridin-3-yl-methanethio-4 - [, N- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3.2 .0) Preparation of hept-2-ene-2-carboxylate 5___

OH

10 f 1) 0 r ^ o π "ρ (0φ) 2 ^ rn n 2) CH - \ CO2PNB 3 \ 20 5 II \ CO2pnb p-nitrobenzyl-3- [1- (R, S) methyl-pyridin-3 -yl 30-methanethiol (N) (R) -hydroxyethyl N, N-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate in a cooled solution (0 ° C) in which was 1.85 g (5.3 mmol) of the keto intermediate 5 in 20 ml of acetonitrile, 754 mg (5.8 35 mmol) of diisopropylethylamine in 1 ml of acetonitrile were added under a nitrogen atmosphere and then a solution of 1.57 g (5.84 mmol) of diphenyl chlorophosphate in 2 ml of acetonitrile. The resulting solution was stirred for 15 minutes at 0 ° C, and then a solution of 754 mg (5.8 to 5 mmol) of diisopropylethylamine in 1 mL of acetonitrile was added, followed by 814 mg (5.8 mmol) of thiol. in acetonitrile. The mixture was stirred at 0 ° for 3 hours, and then the reaction mixture was diluted with 200 mL of ethyl acetate, and washed with ice-cold brine (200 mL), water (200 mL), aqueous bicarbonate (100 mL), and brine (100 mL). Evaporation of the dried (MgSO 4) solution to dryness gave a yellow oil which was purified by column chromatography on silica gel eluting with 50% acetone to 50% methylene chloride to give 1.65 g of the title product as a yellow solid.

NMR (CDCl 3) δ: 1.22 and 1.25 (3H each d, J = 7.0

Hz), 1.46 and 1.50 (3H each d, J = 7.2 Hz), 2.4-3.3 (3H, m), 3.8-4.2 (3H, m), .35 (2H, ABq, J = 14.5 Hz) δ and 7.2-8.6 (8H, m).

IR (KBr) λ max: 3400, 1765 and 1690 cm -1

Analysis calculated for C 23 H 23 N 3 O 3 • 1: C, 58.83; H, 4.94; N, 8.95; S, 6.83; Found: C, 57.15; H, 5.04; N, 8.28; S, 6.78.

25 δη

Ύ * 1 \ / SH

SOCI, Js. 1) $ JL

-Z- * H-nA.NH2

L j TT-I

Z) KACh

HCl 25 £ S 27 82 78094 4- (11-mercaptoethyl) pyridine

To a solution of 25 g of 1- (4-peridyl) ethanol 25 was prepared by the method described in J. Chem. Soc., Perkin II, 1462 (1974) in 100 ml of chloroform, 50 g of thionyl chloride were added.

The mixture was boiled for 2 hours. Evaporation of the solvents in vacuo gave the chloro compound 26 as a semi-solid which was used in the next step without further purification. Thus, to a solution of 25 in 160 ml of ethanol was added a hot solution of 14.4 g of thiourea in 75 ml of ethanol. The reaction mixture was heated at reflux for 18 hours. The ethanol was evaporated off and the residue was dissolved in 100 ml of water and the pH was adjusted to 10 by adding 2-norm. NaOHsa. The mixture was stirred at room temperature for 90 minutes, the pH was adjusted to 6.0 by the addition of 6-norm. HCl and extracted with ether (2 x 200 mL). Evaporation of the dried (MgSO 4) solution to dryness gave a yellow oil which was distilled in vacuo at 5 mm Hg and the 20 fractions at 60-65 ° C were collected to give 11.0 g (38% yield) of pure thiol 27 as a colorless oil.

NMR (CDCl 3) δ: 1.70 (3H, d, J = 6.0 Hz), 2.05 (1H, d, J = 5.8 Hz), 4.20 (1H, t, J = 6 .0 Hz, 5.8 Hz), 7.20 (2H, d, J = 6.2 Hz) and 8.5 (2H, d, J = 6.2 Hz).

83 7 8 0 9 4

)B

CH, r ^ \ i • ch3!

sJ ~ 1 ~~ J

co2pnb / 28 /

OH J

XLPNB NS__ilv 2 \ CH- β 3 29

Pd / C OH H2 J [V 3 ^ N -> / + ^ .1 · 30 84 7 8 0 9 4 3- [1- (RS) -Methyl-N-methyl-pyridin-3-yl-methanethio [- (R) -Hydroxyethyl] -7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate To a solution of 1.1 g (2.34 mmol) of compound 28 In 100 ml of acetone, 10 ml of methyl iodide were added. The reaction mixture was stirred for 18 hours at room temperature. The precipitate was collected by filtration and washed with methylene chloride (10 mL) to give 1.4 g (100% yield) of quaternized pyridine 29 as a yellow powder.

N MR (DMSO-d 6): 1.10 (3H, d, J = 6.5 Hz), 1.62 (3H, d, J = 7.5 Hz), 2.6-4.2 (6H, m), 4.39 (3H, s), 5.42 (2H, ABq, J = 13.6 Hz) and 7.9-9.2 (8H, m).

IR (KBr) max: 3400, 1770 and 1190 cm-1.

Analysis calculated for C24H26N3O6S1J1: C, 47.14; H, 4.29; N, 6.87; S, 5.24; Found: C, 47.19; H, 4.78; N, 6.11; S, 5.41.

To a solution of 1.45 g (2.37 mmol) of compound 29 in 120 mL of tetrahydrofuran and 120 mL of ether-20 was added 120 mL of pH 7.0 buffer followed by 1.5 g of 10% palladium on charcoal. . The mixture was hydrogenated at 310 kPa on a Parr shaker for 60 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 15 mL). The combined filtrate and washings were extracted with ether (2 x 200 mL) and lyophilized to give a yellow solid which was purified on a Cg BONDAPAK (Waters Associates) reverse phase column (50 g) eluting with 5% acetonitrile in water at 55 kPa: n pressure.

Each 20 ml fraction was analyzed by high performance liquid chromatography and fractions with ultraviolet absorption at max. 300 nm and lyophilized to give 200 mg (24% yield) of the title product as a yellow amorphous solid. 35 NMR (D 2 O) δ 1.32 (3H, d, J = 7.0 Hz), 1.63 (3H, d, J = 7.2 Hz), 2.5-4.6 (6H, m ), 4.32 (3H, s) and 8.2-8.9 (4H, m).

85 78094 IR (KBr) ν max: 3400, 1750 and 1590 cm-1.

UV T ^ max. (H 2 O): 296 nm (ε = 7573).

Analysis calculated for c 17 H 20 N 2 O 4 Sl ** 1/2 H 2 O: C, 54.38; H, 5.77; N, 7.46; Found: C, 54.39; H, 5.98; N, 7.68.

)B

Example 13 3- (N-Methyl-N'-benzylimidazol-2-ylmethanethio) -6H- (1-) (R) -hydroxyethyl-7--oxo- Preparation of 1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate_20

S H

HCl 1) H2N - N-Ac CV- —Λ Cv 25 mm, Δ V / \ «C1 41 42 30 86 7 8 0 9 4 N-benzyl-2-mercaptomethylimidazole

To a solution of 3.23 g (13.0 mmol) of the chlorine compound 41 was prepared by the method described in J. Amer. Chem. Soc., 71, 383 (1949), in 80 ml of acetonitrile, 1.72 g (14.5 mmol) of N-acetylthiourea were added. The reaction mixture was heated at reflux for 3 hours. The precipitate was collected by filtration and washed with acetonitrile (10 ml) to give the isothiouronium salt, which was then dissolved in 80 ml of absolute ethanol and heated at reflux for 18 hours under a nitrogen atmosphere. The reaction mixture was cooled to room temperature, condensed in vacuo to a volume of about 30 ml and the precipitate was removed by filtration. Evaporation of the filtrate to dryness in vacuo gave 3.5 g (97% yield) of thiol 42 as a yellow thick syrup.

NMR (CDCl 3) δ: 2.1 (1H, t, J = 4.5 Hz), 3.80 (2H, s), 5.20 (2H, s) and 6.8-7.5 (7H, m).

„“ ”0 '' πΛ = ο ci-pW) 2

0 ÖLPNB

: »“ 1 ”} <*

35 COgPNB

43 • mm * 87 7 8 0 9 4 p-Nitrobenzyl-3- [N-benzylimidazol-2-yl-methanethiol] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo D 2.0J / hept-2-ene-2-carboxylate To a cooled solution (0 °) of 3.03 g of the keto intermediate 5 (8.5 mmol) in 70 ml of acetonitrile was added 1.17 g of nitrogen under protection. g (9.0 mmol) of diisopropylethylamine in 2 ml of acetone tril and then 2.4 g (9.0 mmol) of diphenyl chlorophosphate in 2 ml of acetonitrile. The resulting solution 10 was stirred for 20 minutes at 0 ° C and then a solution of 1.17 g (9.0 mmol) of diisopropylethylamine in 2 mL of acetonitrile was added followed by 4.8 g (15 mmol) of thiol 42 An additional 1.93 g (15 mmol) of diisopropylethylamine was added and the reaction mixture was stirred for 2 hours at 0 ° C. The precipitate was collected by filtration and washed with cold methylene chloride (20 mL) to give 2.5 g (55% yield) of the title product as a white solid.

NMR (DMSO-d 6) 1.23 (3H, d, J = 7.2 Hz), 2.5-4.1

Δ (6H, m), 4.25 (2H, s), 5.20 (2H, s), 5.20 and 5.45 (1H

each, d, J = 14.5 Hz) and 6.9-8.3 (11H, m).

IR (KBr) f'max. 3400, 1775 and 1690 cm-1.

78094 88 J r CO2PNB / "W -ό do2PNB 1+ tH3 44 15

Pd / C

h2 i ^ CO, "I + 2 <* 3 25 45 3- (N-methyl-N'-benzylimidazol-2-ylmethanethio) -6oC- (1-) (R) -hydroxyethyl7-7- oxo-1-aza-bicyclo (3,2,0) hept-2-ene-2-carboxylate

To a solution of 1.76 g (3.3 moles) of compound 43 in 1.1 liters of methylene chloride was added 1.15 mL (13.4 mmol) of methyl fluorosulfonate. The reaction mixture was stirred for 2 hours at room temperature. The reaction mixture 35 89 78094 was concentrated in vacuo to a volume of about 15 mL.

The precipitate was collected by filtration and washed with methylene chloride (10 mL) to give 1.58 g (74% yield) of quaternized imidazole 44 as a white solid.

Δ NMR (DMSO-d 6) δ: 1.15 (3H, d, J = 7.0 Hz), 3.2-4.4 (6H, m), 4.70 and 5.0 (1H each, ABq , J = 10.8 Hz), 5.24 and 5.46 (1H each, ABq, J = 14 Hz), 5.50 (2H, s), and 7.5-8.4 (11H, m) .

IR (KBr) fmax: 3500, 1770 and 1700 cm-1.

Analysis calculated for C 28 H 29 N 4 O 9 S 2 F: C, 51.48; H, 4.47; N, 8.67; S, 10.20; received; C, 51.84; H, 4.52; N, 8.65; S, 9.87.

To a solution of 1.11 g (1.71 mmol) of compound 44 in 100 mL of tetrahydrofuran and 100 mL of ether was added 120 mL of pH 7.0 buffer followed by 1.0 g of 10% palladium -puuhiiltä. The mixture was hydrogenated at 310 kPa on a Parr shaker for 45 minutes. The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 70 ml) and lyophilized to give a yellow powder which was purified on a Cg BONDAPAK (Waters Associates) column (40 g) eluting with 10% acetonitrile in water at 55 kPa: n pressure.

Each 15 ml fraction was analyzed using high performance liquid chromatography and fractions with ultraviolet absorption at 'Xmax' were collected. 300 nm and lyophilized to give 305 mg (43% yield) of the title product as a slightly yellow amorphous solid.

NMR (DMSO) 1.40 (3H, d, J = 7.0 Hz), 2.9-3.4 (3H, m), 3.98 (3H, s), 4.0-4.2 ( 2H, m), 4.23 (2H, broad s), 5.57 (2H, s) and 7.2-7.65 (7H, m).

IR (KBr) / max: 3400, 1760 and 1590 cm -1.

35 UV JV. max (H 2 O); 299 nm (= 8807).

9o 78094

Analysis calculated for C 21 H 23 N 3 O 4 S 1 • H 2 O 2: C, 57.25; H, 5.94; N, 9.54; S, 7.28; Found: C, 56.66; H, 5.70; N, 9.49; S, 8.30.

5 OH

^ prv * £ Q

10 o 3 T ©: o2 © cH3

Example 1/4 3- (2-Methyl-N-methylpyridin-3-yl-methanethio) -15,6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo- (3.2, 0) Preparation of hept-2-ene-2-carboxylate

mp Y'C ° 2 H L AH

L Jl —— * Γ - ^ - ci2> 20 ^ νΛ% YJYm 2? is "m mm

S

Dh.nAnh, rr ^ c1 —► yy ^ sh

N 2 CH, 2> NaOH

HCl 30 15 78094 2-Methyl-3-mercaptomethyl-pyridine

Ester 12 was prepared by the method described in J. Org. Chem., 21, 800 (1956). To a cooled suspended solution (0 °) of 2.86 g of lithium aluminum hydride in 50 ml of dry tetrahydrofuran was added dropwise over 15 minutes a solution of 6.23 g (0.038 mol) of ester 12 in 15 ml of tetrahydrofuran. The mixture was stirred for 60 minutes at 0 °, and then 50 ml of ethyl acetate was added. The precipitate was filtered off and washed with saturated aqueous ammonium chloride solution. The organic layer was dried over MgSO 4, filtered and evaporated to dryness in vacuo to give 3.2 g (70% yield) of hydroxymethylpyridine 13 as a yellow oil. 13 NMR (CDCl 3) δ: 2.46 (3H, S), 4.73 (2H, S), 5.1 (1H, broad), 7.2 (1H, dd, J = 8) Hz), 7.8 (1H, dd, J = 8 Hz, J = 1 Hz) and 8.3 (1H, dd, J = 7 Hz, J = 1 Hz).

To a cooled solution (0 °) of 4 ml of thionyl chloride in 10 ml of methylene chloride was added dropwise over 15 minutes, under a nitrogen atmosphere, a solution of 3.2 g (0.026 moles) of alcohol 13 in 10 ml of methylene chloride. The cooling bath was removed and the reaction mixture was stirred for 3 hours at room temperature. All solvents were evaporated off in vacuo to leave compound 14 as a brown solid which was used in the next step without purification. The crude brown solid was dissolved in 30 ml of absolute ethanol. 2.5 g (0.032 mol) of thiourea were then added and the mixture was heated at 65-70 ° C for 18 hours. The mixture was cooled to room temperature. The precipitate was collected by filtration and washed with ethanol (20 ml) and ether (50 ml) to give 30 g of the isothiouronium salt. This salt was dissolved in 10 ml of water, and 640 mg (0.016 mol) of sodium hydroxide in 10 ml of water were added under nitrogen. The reaction mixture was heated at 100 ° for 2 minutes and then cooled to 0 ° C, adjusted to pH 6.0 with acetic acid and extracted with chloroform (2 x 35 mL). Evaporation of the dried (MgSO 4) chloroform solution to dryness gave 941 mg (46% yield) of thiol as a yellow oil.

Δ Thiol NMR (COCl-j) δ: 1.8 (1H, t), 2.60 (3H, S), 3.73 (2H, d, J = 10Hz), 7.13 (1H, dd, J = 8 Hz), 7.57 (1H, dd, J = 8 Hz) and 8.43 (1H, dd, J = 8Hz, 3Hz).

- T J.

c ° 2pnb cVnb r 5 15 JL 16-uuj *** p-nitrobenzoyl-3- (2-methylpyridin-3-ylmethanethio) -6 * - [1- (R) -hydroxyethyl, 7-7 -oxo-1-20 azabicyclo (3,2,0) hept-2-ene-2-carboxylate To a cooled solution (0 °) of 1.52 g (4.37 mmol) of the keto intermediate 5 in 5 mL acetonitrile, 0.86 ml (4.80 mmol) of diisopropylethylamine was added under a nitrogen atmosphere, followed by a solution of 1.17 g (4.37 mmol) of diphenyl chlorophosphate in 3 ml of acetonitrile. The resulting solution was stirred for 30 minutes at 0 ° C to give p-nitro-benzyl-3- (diphenylphosphoryloxy) -6J- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3, 2.0) hept-2-carboxylate-30 thia. To this solution was added a solution of 0.86 mL (4.80 mmol) of diisopropylethylamine in 2 mL of acetonitrile followed by a solution of 940 mg (6.76 mmol) of thiol in 2 mL of acetonitrile. The reaction mixture was stirred for 60 minutes at 0 ° C. The precipitate was collected by filtration and washed with ether (30 ml), 93 7 8 0 9 4 to give 1.12 g (55% yield) of the title product as a pale yellow solid. Sp. 186-188 ° C (dec.).

NMR (DMSO-d 6): 1.20 (3H, d, J = 7 Hz), 2.60 δ (3H, S), 3.40 (ra, 2H), 4.16 (m, 2H), δ , 32 (2H, S), 5.16 (1H, d, J = 5 Hz), 5.44 (2H, q, J = 14 Hz), 7.32 (2H, m), 7.8 (2H , d, J = 8 Hz), 8.36 (2H, d, J = 8 Hz) and 8.48 (1H, dd, J = 5.5 Hz, 1.5 Hz).

IR (KBr) ν max: 3500, 1770 and 1750 cm -1 Analysis calculated for C 23 H 24 N 3 O 6 S: C, 58.83; H, 4.94; N, 8.94; S, 6.83; Found: C, 58.63; H, 4.99; N, 9.06; S, 6.58.

OH

15 J- Γ CH, N 1 1 Θ 20 0 CO2pNB tH3 FSO3 © 16 17 25 Pd / C, H2 OH ^

ci Ö c £ Y

1 © 18 CH3 35 94 780 9 4 3- (2-Methyl-N-methylpyridin-3-yl-methanethio) -6H --- (R) -hydroxyethyl-7--oxo-1-azabicyclo- (3.2.0 = hept-2-ene-2-karboksylaatti_

To a solution of 697 mg (1.19 mmol) of compound 16 in 100 mL of methylene chloride was added dropwise at 10 ° C over 0.5 min, 0.5 mL (6.18 mmol) of methyl fluorosulfonate. The mixture was stirred for 2.5 hours at room temperature. The precipitate was collected by filtration and washed with 30 mL of methylene chloride to give 777 mg (90%) of quaternized pyridine 17 as a yellow solid.

NMR for 17 (COCl 3) δ: 1.20 (3H, d, J = 7 Hz), 2.82 (3H, s), 4.36 (3H, s), 4.16 (2H, m), 4 , 60 (2H, s), 5.20 (1H, m), 5.42 (2H, q, J = 14 Hz), 7.80 (2H, d, J = 8 Hz), 8.04 ( 1H, dd, J = 7 Hz, 6.5 Hz), 8.32 (2H, d, J = 8 Hz), 8.64 (1H, d, J = 7.5 Hz) and 9.08 (1H , d, J = 7.5 Hz).

IR (KBr) λmax: 3500 and 1765 cm -1 Analysis calculated for C 24 H 26 FN 3 O 9 S: C, 48.91; H, 4.55; N, 7.23; S, 11.04; Found: C, 49.39; H, 3.97; N, 7.20; S, 10.98.

To a solution of 1.10 g (1.88 mmol) of compound 17 in 80 mL of tetrahydrofuran and 80 mL of ether was added 80 mL of pH 7.0 buffer followed by 800 mg of 10% palladium on charcoal. The mixture was hydrogenated at 25,207 kPa on a Parr shaker for 40 minutes.

The mixture was filtered through a pad of Celite and the catalyst was washed with water (2 x 10 mL). The combined filtrate and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow powder which was purified by HP-20 column chromatography eluting with water followed by 5% acetonitrile. Each 15 ml fraction was analyzed using high pressure liquid chromatography and fractions with ultraviolet absorption at Xmax. 300 nm, combined and lyophilized to give 614 mg (42% yield) of the title product as a slightly yellow powder.

95 78094 NMR (D 2 O) δ 1.28 (d, 3H, J = 7 Hz), 2.86 (3H, s), 3.20 (2H, dd, J = 10 Hz, 3.5 Hz), δ , 42 (1H, dd, J = 5.4 Hz, 3.5 Hz), 4.20 (3H, m), 4.32 (3H, s), 4.35 (2H, S), 9.88 (1H, dd, J = 7.2 Hz, 6.5 Hz), 8.5 (1H, d, J = 8 Hz) δ and 8.70 (1H, d, J = 8 Hz).

IR (KBr) ν max: 3400, 1760 and 1590 cm -1.

UV a.max. (H 2 O): 298 nm (€ = 8391).

Analysis calculated for C 17 H 20 N 2 O 3 S.H 2 O: C, 55.73; H, 5.46; N, 7.65; S, 8.74; Found: C, 55.50; H, 6.05; M, 7.74; S, 8.68.

Example 15 3- [4- (Ν, Ν-Dimethyl-1,2,3-triazolium) -methyl]? Preparation of 6 - [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo (3,2,0) hept-2-ene-2-carboxylate

OH

p / VvCH3

20 J - + ^ N

0 Λο, * A. Preparation of isomer A 25 AH?

y SKc a) MeOTf ^ 1 — Γ ^ \ ^ XX jT

\ b) Aq. NaOH \ I J, // T ^ \ Γ C OH 0 ^^ CO ® / m, N-h 2 / φ • i A. g> o Me

We

€ 0-PNB

35 2

d) H2 / Pd-C

96 7 8 0 9 4

Methyl trifluoromethanesulfonate (0.58 mL, 5.16 mmol) was added dropwise under ice-protection to an ice-cold, stirred solution of 4- (methanethiol acetate) -1-methyl-1,2,3-triazole (590 mg, 5 3.52 mmol) in dry methylene chloride (2 mL). After 0.5 h, the bath was removed and after 1 h, the solvent was removed by suction. The residual oil was dissolved in a few milliliters of water and this solution was frozen in an ice bath. A cold solution was then added if sodium hydroxide (305 mg, 7.59 mmol) was added to the other mass per milliliter of water and the reaction mixture was stirred for 0.75 hours. The solution was diluted to 25 ml with water and the pH was adjusted to 7.5 by the addition of solid sodium dihydrogen phosphate monohydrate. 14 mL of this solution (about 1.9 mmol of triazolium thiol) was then added to an ice-cooled, stirred solution of enol phosphate (1.0 g, 1.72 mmol) in tetrahydrofuran (THF) (10 mL). This was stirred for 0.75 hours (during this reaction some crystalline material precipitated, probably Na 2 HPO 4). The suspension was transferred to a pressure flask with the aid of some THF (20 mL) and water (20 mL). Ether (30 ml) and 10% palladium on charcoal (1.0 g) were added and the mixture was hydrogenated (276 kPa) for one hour. The organic phase was separated and washed with water (2 x 5 mL). The combined aqueous phases were filtered and the filtrate was concentrated under high vacuum (ca. 0.5 mm, 1.5 h). The yellow solution was then chromatographed (medium pressure reverse phase column, 35 x 90 mm, H 2 O as eluent) to give, after freeze-drying, 395 mg of carbepenem with some inorganic material as a minor impurity. The product was purified by HPLC (10 x 300 mm Waters Microbondapack C-18 column, multi-jet, H 2 O as eluent) to give 310 mg (57%) of isomer A 35 as a tan powder: δ 78094 1 HNMR (D 2 O) δ: 1.23 (3H, d, J = 6.4 Hz), 3.10 (2H, d, J = 9.1 Hz), 3.24 (1H, q, J = 2.7, 6.1 Hz), 4 3.04.71 (10H, m), 8.46 (1H, s); IR (nujol) 1760 cm-1; UV (phosphate buffer, pH 7.4, M = 0.05) λmax: 296 (¢. = 7,500).

B. Preparation of Isomer B and Isomer C.

OH

a) MeOTf XH H

10 ^ SAc b) aq. NaOH t \ ALLv C ° 2® (hypothetical structure I_ ^ Λ — Oj *, γ) O 0

C02PNB

d) H2 / Pd-C

Methyl trifluoromethanesulfonate (1.60 mL, 14.0 mmol) was added dropwise under ice-protection to an ice-cooled solution of 4- (methanethiol acetate) -2-methyl-1,2,3-triazole (1.20 g, 7, 02 mmol) in dry methylene chloride (6 mL). This was allowed to warm to room temperature and stirred for 16 hours. Additional methyl trifluoromethanesulfonate (0.40 mL, 3.56 mmol) was added and after 3 h at room temperature the solvent was removed in vacuo. The residual oil was triturated with ether and the resulting resin was dissolved in water (5 mL). This was cooled in an ice bath and a solution of sodium hydroxide (844 mg, 21.1 mmol) in water (5 mL) was added. After stirring for 0.75 hours, this solution was diluted to 60 ml with water and the pH was adjusted to 8 by the addition of solid potassium dihydrogen phosphate. 40 mL of this solution (about 4.7 mmol of a mixture of triazolium thiol isomers) was then added to an ice-cooled, stirred solution of 98 7 8 0 9 4 enol phosphate (2.00 g, 3.45 mmol) in THF (60 mL).

This mixture was stirred in an ice bath for 0.5 h, then transferred to a pressure flask containing a suspension of 10% palladium on charcoal (2.00 g) and ether (60 mL). The mixture was hydrogenated (276 kPa) for one hour.

The organic phase was separated and washed with water (2 x 10 mL). The combined aqueous phases were filtered and the filtrate was concentrated under high vacuum (ca. 0.5 mm, 1.5 h). The remaining solution was then chromatographed (medium pressure 10 reversed-phase column, 45 x 130 mm, 1 x 0 as eluent) to give, after freeze-drying, 595 mg of a mixture of carbapenem isomers with a small amount of inorganic impurity. These were separated and purified by HPLC (10 x 300 mm Waters 15 Microbondapack C-18 column, multiwell, eluent 1 → 0) to give, in the order of elution: isomer B: 153 mg (13%); 1 HNMR (D 2 O) 1.23 (3H, d, J = 6.4 Hz), 3.12 (2H, q, J = 1.4, 8.9 Hz), 3.39 (1H, q, 3 = 2.1, 6.0 Hz), 4.07-4.68 (10H, m), 8.19 (1H, s); IR (nujol) 1755 cm-1; UV (phosphate buffer, pH 7.4, M = 0.05) Amax. 296 nm (ε = 6'70 °)? and Isomer C: 284 mg (24%); 1 HNMR (D 2 O) δ: 1.23 (3H, d, H = 6.4 Hz), 3.15 (2H, q, 25 J = 3.7, 9.0 Hz), 3.37 (1H, q , J = 2.6, 6.0 Hz), 3.95-4.65 (10H, m), 8.62 (1H, s); IR (nujol) 1750 cm -1; UV (phosphate buffer, pH 7.4 M = 0.05) λmax 5 298 nm <ε = 7'60 °) · I: 99 7 8 0 9 4

Example 16 (5R, 6S) -6- (1R-Hydroxyethyl) -3- (2-methyl-1,2,3-thiadiazolium-4-ylmethylthio) -7-oxo-1-azabicyclo [5 # 2 # OJ7hept-2-ene-2-carboxylate__ 5

OH

Λπ-νι ΊΗ \ ®

10 0 VP

A. Ethyl 1,2,3-thiadiazol-4-ylcarboxylate. f -J ”»> r \ c »3 / Ng ° Et \ · ^ 20

A solution of ethyl N-carbethoxyhydrazonopropionate (31.2 g, 0.154 mol) in thionyl chloride (80 mL) was stirred at 23 ° C for 3 hours and heated at 70 ° C for 20 minutes. The thionyl chloride was evaporated and the residue was triturated with hexane (4 x 30 ml). The red solid was dissolved in dichloromethane (150 mL) and the solution was washed with saturated sodium bicarbonate solution and water. After drying over Na 2 SO 4, the solution was concentrated until the compound crystallized. After the mixture was stationary at 23 ° C for some time, the crystals were filtered off; 16.8 g, m.p. 86 ° C, 69%. The filtrate was concentrated and purified by silica gel column chromatography with dichloromethane as the eluting solvent to give 3.17 g of product, 35 m.p. 86 ° C, 13%; 100 78094 IR (KBr) δ max: 1720 (ester) cm -1; 1 Hmr (CDCl 3) δ: 1.52 (3H, t, J = 7.1 Hz, CH 3 CH 2 O), 4.57 (2H, q, J = 7, l

Hz, CH 2 -CH-O), 9.47 (1H, s, thiadiazoline H).

CD. Hurd and R.I. Mori, J. Am. Chem. Soc., 77, 5359 5 (1955).

B. 1,2,3-Thiadiazol-4-ylmethanol1

„.“ CH_OH

fiOEt _ / 2 Q -. 0

To a suspension of ethyl 1,2,3-thiadiazol-4-ylcarboxylate (18.35 g, 0.116 mol) in ether 15 (400 mL) was added lithium aluminum hydride (2.47 g, 0.065 mol) portionwise over 1 hour. The reaction mixture was stirred at 23 ° C for 7 hours and treated with lithium aluminum hydride (2.47 g, 0.065 mL). Stirring was continued for 24 hours before hydrogen (7 ml), 15% sodium hydroxide solution (7 ml) and water (21 ml) were added sequentially. After stirring for 15 minutes, the ether solution was decanted off and the resin was extracted with ether (5 x 100 mL). The ether extracts were combined, dried (MgSO 4) and concentrated (5.4 g). The crude material was purified on a silica gel column (120 g, 4 x 16 cm) using ether as the eluting solvent to give 1.3 g (7%) of ethyl 1,2,3-thiadiazol-4-ylcarboxylate and 2.45 g. g (18%) 1,2,3-thiadiazol-4-ylmethanol; IR (film) ν max: 3380 (OH) cm -1; 1 Hmr (CDCl 3) 2.31 (1H, s, OH), 5.22 (2H, s, CH 2 O), 8.50 (1H, s , thiadiazoline H).

1S.I. Rämsby, S.0. ögren, S.B. Ross and N.E. Stjernström, * Acta Pharm. Succica., 10, 285-96 (1973); C.A., 79, 35 137052W (1973).

101 78094 C. 1,2,3-Thiadiazol-4-ylmethanol methanesulfonate_ XH OH_ ^ HOXs rt i) 3 NEt3 5

A solution of 1,2,3-thiadiazol-4-ylmethanol-10 L (0.75 g, 6.5 mmol) in dichloromethane (20 mL) was cooled to 5 ° C under a nitrogen atmosphere and treated with triethylamine (1.018 mL). , 7.3 mmol) and methanesulfonyl chloride (0.565 mL, 7.3 mmol). After 15 minutes, the ice bath was removed and the reaction mixture was stirred for 2 hours.

The solution was washed with 1-norm. hydrochloric acid solution (2 x 2 mL) and water, dried (MgSO 4 + MgO) and concentrated. The residue was purified by chromatography (silica gel column 1.5 x 21 cm) using ether as the eluting solvent to give 0.90 g (71%) of 1,2,3-thiadiazol-4-ylmethanol methanesulfonate; and (film) λ max: 1350 (SO 2) cm -1, 1172 (SO 2) cm -1; 1 Hmr (CDCl 3) ε; 3.09 (3H, s, CH 3), 5.75 (2H, s, CH 2) 8.72 (1H, s, thiadiazole H); uv (CH 2 Cl 2 Xmax: ^ 1 25 (£ 1990); analysis calculated for C 9 H 9 N 2 O 3 S: C, 24.73; H, 3.11; N, 14.42; S Found: C, 24.78; H, 3.09; N, 14.66; S, 31.94; and 0.13 g (19%) of di- (1,2,3-thiadiazole); 4-ylmethyl) ether-30 blades; ir (film) λ max: 1272, 1242, 1200, 986, 805, 728 cm -1; 1 Hmr (CDCl 3) δ ·: 5.16 (s, 4H, CH 2), 8.42 (s, 2H, thiadiazoline Hs).

102 78094 D. 4-Acetylthiomethyl-1,2,3-thiadlazole 5 ^ CHOMs 9 „_x'CHsEch η" ·. Ιί x s' s

To a solution of 1,2,3-thiadiazol-4-ylmethyl-10-methanol methanesulfonate (0.90 g, 4.6 mmol) in tetrahydrofuran (9 mL) was added sodium thiolacetate prepared from thiolacetic acid (0.38 mL, 5.3 mmol) and aqueous sodium bicarbonate (0.445 g, 5.3 mmol) (2 mL). The resulting mixture was stirred at 23 ° C for 15 h and diluted with ether (75 mL). The organic solution was washed with water (3 x 3 mL), dried (MgSO 4) and concentrated. The crude mixture was purified by chromatography (silica gel column: 1.4 x 19 cm) using 50% ether in hexane as eluent to give 0.60 g (75%) of product; and (film) ν max: 1675 (C = O) cm -1; 1 HMr (CDCl 3) δ ·. 2.37 (3H, s, CH 3), 4.58 (2H, s, CH 2), 8.44 (1H, s, thiadiazoline H).

Analysis calculated for O 2 O 2 O 2 032: 25 C, 34.47; H, 3.47; N, 16.08; S, 36.80; Found: C, 34.48; H, 3.83; N, 16.28; S, 36.80.

i.

103 7 £ 0 94 E. 4-Acetylthiomethyl-2-methyl-1,2,3-thiadiazole-trifluoromethanesulfonate and 4-acetylthiomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate_5

CH s? CH / H S ^ CH

, / 2 3 cF3SO3Me K- ^ 10 - ^ <- ·%. / ^ 3 ^ 3 /

\ _ ^ CH sicH

Ö -Λ: 15

To a solution of 4-acetylthiomethyl-1,2,3-thiadiazole (0.60 g, 3.44 mmol) in a mixture of ether (4 mL) and dichloromethane (0.4 mL) was added a few crystals of the title compound and trifluoromethanesulfonate. (0.407 mL, 3.6 mmol) over 5 minutes. The reaction mixture was stirred at 23 ° C under a nitrogen atmosphere for 6 hours. The white solid, a mixture of the two title compounds, was filtered off and washed with ether; 1.05 g, 90%, 25 and (KBr) m.p. 1675 (C = O) cm -1 Hmr (DMSO, d-6) g: 2.43 (3H, s, Cl 2 COS), 3.33 (s, CH 2 in N-3), 4.57 ( s, in CH 3 N-2), 4.66 (2H, s, CH 2), 9.55 (H in thiadiazolium N-2, 9.66 (H in thiadiazolium N-3)).

Analysis calculated for C 18 H 9 N 2 O 2 S 2 F 3: C, 20.27; H, 2.38; N, 9.45; S, 32.46; Found: C, 24.61; H, 2.57; N, 8.47; S, 28.21.

104 78094 F. 4-Mercaptomethyl-2-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate and 4-mercaptomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate_ 5

XH S? CH

2 3> CH SH

^) - —cl * 6N> Me ^ + 1

CF SO »N U CF SO

10 XS 3 3

A solution of 4-acetylthiomethyl-2-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate and a mixture of 4-acetylthiomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate (1.05 g, 3, 1 arrow) 6-norm.

In 15 hydrochloric acid (10 ml) was heated at 65 ° C under a nitrogen atmosphere for 1.75 hours. The solvent was evaporated in vacuo to leave a yellow syrup of 0.91 g. This compound was used in the next step without purification.

20 105 78094 G. (5R, 6S) -6- (1R-hydroxyethyl) -3- (2-methyl-1,2,3-thiadiazolium-4-ylmethylthio) -7-oxo-1-azabicyclo [5.2 .0 / hept-2-ene-2-carboxylate

Λγ-λ,? V

J-T - 7> ίο ° Λοο, κβ

kf '0H

10% Pd / c "

ethers, Cr ^ 'Cr II V — M

15 HO * - ^

13 2 COO J

Cold solution (5 ° C) of (5R, 6S) -paranitro-benzyl-6- (1R-hydroxy-ethyl) -3- (diphenylphosphono) -7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate-20 (1.7 g, 2.92 mmol) in tetrahydrofuran (10 mL) was treated with a solution of crude 4-mercaptomethyl-2-methyl-1,2, A mixture of 3-thiadiazolium trifluoromethanesulfonate and 4-mercaptomethyl-3-methyl-1,2,3-thiadiazolium trifluoromethanesulfonate (0.9 g) in phosphate-25 buffer (pH 7.2, 0.5 mol), 15 ml) and tetrahydrofuran (5 ml). The reaction mixture was stirred for one hour and the pH was maintained at 7.2 2-norm. solution of sodium-chloride solution. Stirring was continued for an additional hour before ether (50 mL) and 10% palladium-on-carbon (1 g) were added. The resulting mixture was hydrogenated at 23 ° C under 310 kPa for 2 hours and filtered through a pad of Celite. The organic phase is separated, diluted with ether (50 ml) and phosphate buffer (pH 7.2, 0.3 mol, 20 ml) and hydrogenated (2 g of 10% palladium on charcoal) for 2 hours at 345 kPa. under. The aqueous phases were combined (from the first and second hydrogenolysis, washed with ether and purified by chromatography on PrepPak 500-C / 18 with water as eluent to give 0.22 g of crude material.

It was re-purified using hplc eluting with water to give 0.040 g (4%) of the title compound after freeze-drying; ir (KBr) Y> max: 3400 (br, OH), 1745 (C = O-lactam)), 1580 (carboxylate) cm -1 H 2 Hmr (D 2 O) 1.23 (3H, d, J = 6.3 Hz, CH 3 CHOH, 3.04, 3.05, 3.16 (2H, m, 10 H-4), 3.38 (1H, dd, J = 2.8 Hz, J = 6.0 Hz , H-6), 3.9-4.6 (2H, m, H-5, CH 3 CHOH), 4.51, 4.53 (2 "s", SCH 2), 4.61 (s, N + CH 3 uv (H 2 O) λ max>: 224 (E = 4.345), 262 (E = 4980), 296 (i = 6885), 18 ° (c 0.18, H 2 O); λ / 2 = 9.8 h (measured at a concentration of 10 .mu.mol.

In phosphate buffer pH 7.4 at 36.8 ° C).

107 7 8 0 9 4

Example 17

Potassium 3- [4- (1-carboxylate methyl-3-methyl-1,2,3-triazolium) -methanethio] - [4- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo] ^, 2,0 / hept-2-ene-2-5 carboxylate-co ^ SH HH 2 Λϊ-4-Λ ^ ^ 9 acs ^ Ntn ^ ΗΟά ^ Ν 1) 1AH 'nt,> - * S' | 2) MsCl / NEt ^ * kscoch3 «

BrCH ^ OEt CO © ^ © ^ Et0 * H H nT 2 AcS ^ Y »^ Br ^

<_r, _ V

J— nI / K0H I

0 β ^ n / OH

CO, J 1HH h o bl

COPNB

2

c) H2 / Pd-C

108 7 8094

Lithium aluminum hydride (2.83 g, 70.9 mmol) was added in small portions to a stirred suspension of 1-methyl-1,2,3-triazole-4-carboxylic acid (9.00 g, 70.9 mmol) in dry THF. (200 ml). Mixture 5 was stirred at room temperature for 15 hours, after which 20% aqueous sodium hydroxide solution (20 ml) was carefully added in about 1 ml portions. The resulting granular suspension was filtered and the solid was washed with added THF (5 x 75 mL). the combined THF solutions were dried (MgSO 4) and the solvent was removed. The residual yellow oil was flash chromatographed on a silica gel column (90 x 35 cm) eluting with 100 ml portions of hexane, ethyl acetate-hexane (1: 1) and (1: 3), and finally ethyl acetate-methanol. mixture (9: 1) ^. There was thus obtained 4-hydroxymethyl-1-methyl-1,2,3-triazole (3.18 g, 40%) as a colorless oil; 1 HNMR (CDCl 3) δ: 4.07 (3H, s), 4.73 (2H, d), 7.52 (1H, s); IR (alone) 3320 cm

Methanesulfonyl chloride (3.82 mL, 49.6 mmol) was added dropwise to an ice-cold stirred solution of alcohol (4.67 mL, 41.3 mmol) and triethylamine (7.47 mL, 53.7 mmol) in methylene chloride ( 20 ml). After 0.5 h, the solvent was removed and the remaining solid was dissolved in acetonitrile (25 mL). Potassium thiol acetate (7.06 g, 62.0 mmol) was then added and the suspension was stirred at room temperature for three hours. Additional potassium thiol acetate (3.0 g; 26.3 mmol) was added and the suspension was further stirred for 16 hours. The dark colored suspension was then concentrated and water (10 mL) was added. This mixture was extracted with methylene chloride (5 x 40 mL). The combined extracts were dried (MgSO 4) and the solvent removed. The residual oil was flash chromatographed on a silica gel column (90 x 36 mm) using 35 ° C. Pederson, Acta. Chem. Scand., 1959, 13, 888.

109 78094 with hexane as eluent followed by hexane-ethyl acetate (1: 1). This gave 4- (methane-thiolacetate) -1-methyl-1,2,3-triazole (5.95 g, 84%) as a pale pink solid: δ 1HNMR (CDCl3) δ: 2.40 (3H, s). , 4.10 (3H, s), 4.20 (2H, s), 7.53 (1H, s); IR (nujol rate) 1675 cm -1.

A solution of triazole (1.00 g, 5.85 mmol) and ethyl bromoacetate (1.48 mL, 13.3 mmol) in dry acetonitrile (10 mL) was heated at 60 ° C for 90 h under nitrogen. . The solvent was removed and the residual oil was triturated with ether (4 x 25 mL) to leave 1-methyl-3- (ethylcarboxymethyl) -4-methanethiol acetate-1,2,3-triazole-15 umbromide as a brownish resin which was used. as such.

To an ice-cooled stirred solution of triazolium bromide in water (20 mL) was added a cold solution of KOH (0.66 g, 12 mmol) in water 20 (5 mL). After 20 minutes, this was diluted to 35 ml and a sufficient amount of solid potassium dihydrogen phosphate was added to bring the pH of this solution to 8.0. This was then added to a stirred, ice-cooled solution of enol phosphate in THF (35 mL).

After 0.5 h, this mixture was transferred to a pressure flask containing ether (35 mL) and 10% palladium on charcoal (1.5 g). Hydrogenated at 276 kPa for 55 minutes. The organic phase was then separated and washed with water (2 x 5 mL). The combined aqueous phases were filtered and the filtrate was concentrated under high vacuum. The residue was chromatographed on a reverse phase column (35 x 120 ml) with water as eluent. Freeze-drying of the carbapenem-containing fractions left 1.20 g of a green-colored solid. This was rechromatographed on Waters Prep. 500 on an HPLC column (PrepPaK-500 / C ^ g- no 78094 column) with 2% acetonitrile in water as eluent. Fractions containing carbapenem were combined and lyophilized. This material was rechromatographed by HPLC (10 x 300 mm Waters 5 Microbondapack C-18 column) with water as eluent to give, after lyophilization, the pure title compound (190 mg, 17%) as a pale yellow solid: 1 HNMR (D 2 O): 1 , 24 (3H, d, J = 6.4 Hz), 3.07 (2H, d, 10 J = 9 Hz), 3.38 (1H, q, J = 2.7, 6.0 Hz), 4.02-4.30 (3H, m), 4.29 (3H, s), 5.23 (2H, s), 8.52 (1H, s); IR (nujol phase) 1750 cm UV (phosphate buffer, pH 7.4) λmax: 296 nm (ε = 7,520)

Example 18 Potassium 3- [4- (1-carboxylate methyl-3-methyl).

1,2,3-triatsolium) metaanitio7 "6 ^ - / L- (R) -hydroxy. Ethyl 7-oxo-1-azabicyclo [3.2.0] hept-2-ene> 2-carboxylate

„0H

C02 0 CO ^ 25 H0? - = H0C X .__ CO-Et HO '' ~ S) = \ ♦ - »O-7 -» Ow 'a) EtOCCl, NEt3 N ^ 2 **

30 EtoicH2N3 b KaBH

4> - £ »., 2 ΡΦ3 35“ P * ▼ ιη 78094 OH ru T H H, 3 V \ © ^ n /

c ° 2 J

L_ 10 a) MeOTf /

b) KOK OH

<=> CO PNB 2

15 d) H2 / Pd-C

A mixture of ethyl azidoacetate (30.0 g, 0.23 mol) and propiolic acid (14.3 ml, 0.23 mol) in toluene (75 ml) was stirred at room temperature.

The reaction remained slightly exothermic for 1.5 hours, after which it rapidly became strongly exothermic and cooling in an ice bath was necessary. At the end of this exothermic step, the reaction mixture was heated at reflux for 0.5 hours. After cooling in an ice bath, the crystalline material was collected by filtration and washed with a small amount of toluene. The crude material thus obtained (33.3 g, 72%) was one isomer 1 HNMR (DMSO-d 6) δ · 1.20 (3H, t, J = 7 Hz), 4.15 (2H, q, J = 7 Hz), 5.42 (2H, s), 8.67 (1H, 3) J, according to previous work, probably 1- (ethylcarboxymethyl) -1,2,3-triazole-4- carboxylic acid.

^ C. Pederson, Acta. Chem. Scand., 1959, 13, 888 112 78094

A solution of carboxylic acid (5.00 g, 25.1 mmol) and triethylamine (3.68 mL, 26.4 mmol) in dry methylene chloride (50 mL) was added to an ice-cooled, stirred solution of ethyl 5-chloroformate ( 2.52 mL, 26.4 mmol) in dry methylene chloride (50 mL). The purple solution was stirred for 0.5 h, then washed with water (10 mL), dried (MgSO 4) and the solvent removed. The crude mixed anhydride was dissolved in THF (50 mL) and added slowly to an ice-cold suspension of sodium borohydride (0.72 g, 18.9 mmol) in THF (50 mL). After stirring for 0.5 h, more sodium borohydride (0.30 g; 7.9 mmol) was added and the reaction mixture was left in an ice bath for 1 h. Vet (15 mL) was then added and after 10 minutes 10% aqueous HCl (3 mL) was added. After gas evolution ceased, solid potassium carbonate (2 g) was added with stirring. The organic phase was then removed and the remaining white paste was extracted with added THF. The combined organic phases were dried (MgSO 4) and the solvent removed. Flash chromatography on a silica gel column eluting with hexane, ethyl acetate-hexane, and finally ethyl acetate gave 1- (ethylcarboxymethyl) -4-hydroxymethyl-1,2,3-triazole-25 (2.04 g). , 44%) as a crystalline solid: 1 HNMR (CDCl 3) δ: 1.28 (3H, t, J = 7 Hz), 4.24 (2H, q, J = 7 Hz), 4.75 (2H, s ), 4.85 (2H, s), 7.73 (1H, s).

Diisopropylazodicarboxylate (4.11 mL, 20.8 mmol) was added dropwise to an ice-cold solution of triphenylphosphine (5.47 g, 20.8 mmol) in dry THF (100 mL) under nitrogen. After 0.5 h, an ice-cold solution of alcohol (1.93 g, 10.4 mmol) and thiolacetic acid (1.49 mL, 20.8 mmol) in dry THF 35 (50 mL) was added to this mixture under nitrogen. This mixture was allowed to stand for 2 hours in an ice bath and then for another 12 hours at room temperature; after which the solvent was removed. The reaction mixture was flash chromatographed on silica gel (40 g, eluting with 100 mL portions of ethyl acetate-hexane mixtures containing 5%, 10%, 15% ... 50% ethyl acetate).

Fractions containing thiol acetate were combined and rechromatographed on silica gel (60 g) / eluting with 200 mL portions of hexane, 5%, 10%, 15%, 20% ethyl acetate-hexane-10 ni mixtures and 22.5% , 25%, 27.5% to 35% ethyl acetate-hexane mixtures).

There was thus obtained 1.24 g (49%) of 1- (ethylcarboxymethyl) -4-methanethiol acetate-1,2,3-triazole as a crystalline solid. 1 HNMR S 1.28 (3H, t, J = 7 Hz), 2 , 37 (3H, s), 3.87 (2H, s), 3.90 (2H, q, J = 7 Hz), 5.12 (2H, s), 7.63 (1H, s); IR (nujol phase), 1735, 1780 cm and in addition 1.40 g of a substance containing triphenylphosphine oxide as an impurity.

Methyl trifluoromethanesulfonate (0.51 mL, 4.53 mmol) was added dropwise to an ice-cold, stirred solution of triazole (1.00 g, 4.12 mmol) in dry methylene chloride (5 mL). The bath was removed after 0.5 h and then, again after 0.5 h, the solvent was removed in vacuo. A white solid remained which was suspended in water (15 ml) and this stirred mixture was cooled in an ice bath. A solution of KHH (0.69 g, 12.4 mmol) in water (5 mL) was added and the reaction mixture was stirred for 1 h. The diluent was then taken up in 30 ml of water and solid potassium dihydrogen phosphate was added to adjust the pH to 8.0. An aliquot of this solution (22 mL, about 3.0 mmol of thiol carboxylate) was added to an ice-cold, stirred solution of enol phosphate (1.60 g, 2.76 mmol) in THF (30 mL). After 0.5 h, the reaction mixture was treated with 78094 114 high vacuum to remove THF. The yellow solution was then chromatographed on a reversed phase column (35 x 120 mm) eluting with water (300 ml) followed by 100 ml aliquots of 5, 10, 15-30% acetonitrile-5-acetonitrile-water mixtures. Freeze-drying the desired fractions gave p-nitrobenzyl ester as a yellow solid (930 mg). This was transferred to a pressure flask containing ether (25 mL), THF (25 mL) and phosphate buffer / 25 mL, prepared by dissolving potassium dihydrogen-10 phosphate (1.36 g, 0.01 mol) in water (100 mL). and adjusting the pH to 7.4 by adding 45% aqueous solution of Κ0Η and 10% palladium on charcoal (900 mg).

The hydrogenation was carried out at 276 kPa for 1 hour, after which the organic phase was separated and washed with water (2 x 5 ml). The combined aqueous phases were filtered and then concentrated under high vacuum. The remaining solution was chromatographed on a reversed phase column (35 x 120 mm) eluting with water. The carbapenem-containing fractions were combined and lyophilized to give 1.21 g of a light green solid. This was then purified by HPLC (10 x 300 mm water-microbondapack C-18 column, eluent H 2 O) to give pure title product, 480 mg (41%): 1 HNMR (D 2 O) g: 1.23 (3H, d, J = 6.4 Hz), 3.11 (2H, d, 25 J = 9 Hz), 3.37 (1H, q, J = 3.0, 6.1 Hz), 4.02 (7H, m ), 5.18 (2H, s), 8.53 (1H, s); IR (nujol phase) 1750 cm UV (phosphate buffer, pH 7'4) Λ max 205 nm <£ = 7.810).

Example 19 3- [5- (1,4-Dimethyl-1,2,4-triazolium) methane (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.1] Hept-2 -ene-2-carboxylate__ 78094 115

OH

JS

1-methyl-5-methanethiolacetate-1,2,4-triassilium__

10 r - N OH a) MsCl / NEt ^ ~ N \\ / AC

/ yj -3 — f \ N> - ^ b) CH3gSH / NEt3 |

Methanesulfonyl chloride (0.46 mL, 6.0 mmol) was added dropwise to an ice-cold stirred solution of 1-methyl-5-hydroxymethyl-1,2,4-triazole * (565 mg, 5.0 mmol) and triethylamine (0 , 91 mL, 6.5 mmol) in methylene chloride (5 mL). After 20 minutes, more triethylamine (1.05 mL, 7.5 mmol / 20 L) was added followed by thiolacetic acid (0.53 mL, 7.5 mmol) and stirring was continued for 45 minutes. The reaction mixture was then diluted with methylene chloride and washed with water. The aqueous phase was extracted with methylene chloride (3 x 5 mL) and the combined organic phases were dried (MgSO 4) and the solvent removed. Chromatography on a silica gel column gave pure 1-methyl-5-methanethiolacetate-1,2,4-triazole (570 mg) as a yellow oil, and the crude fraction (200 mg) was rechromatographed (preparative TLC, silica gel) to give an additional 100 g. mg of pure substance (total yield: 85%) /: 1 HNMR (CDCl 3) δ: 2.38 (3H, s), 3.90 (3H, s), 4.25 (3H, s), 7.80 ( 1H, s).

35 * R.G. Jones and C. Ainsworth, J. Amer. Chem. Soc., 1955, 77, 1938.

116 5 78094 B. 3- [5- (1,4-Dimethyl-1,2,4-triazolium) -methane] -6- [1- [1- (R) -hydroxyethyl] -7-oxo-1 -aza-bicyclo / ,23,2,0J7hept-2-ene-2-carboxylate_ a) MeOTf * Hh u (T “8 / Ac b> NaOH), - '-ÖH-> TTVs / yf

JlO “° - <C> 5.2 lo ©

10 cO PNB

d) H2 / Pd-C

Methyl trifluoromethanesulfonate (1.20 mL, 10.7 mmol) was added dropwise to an ice-cold solution of 1-methyl-5-methanethiol acetate-1,2,4-triazole (730 mg, 4.27 mmol) in methylene chloride (7 mL). The reaction mixture was allowed to slowly warm to room temperature over 3 hours, after which it was concentrated. The residual oil was triturated with ether to leave crude 1,4-dimethyl-5-methanethiol acetate-1,2,4-triazolium trifluoromethanesulfonate (1.46 g) which was used as such.

A solution of sodium hydroxide (512 mg, 12.8 25 mmol) in water (5 mL) was added to an ice-cold solution of the triazolium salt (1.45 g, 4.35 mmol) in water (5 mL). After 45 minutes, this was diluted to 25 ml with water and the pH was adjusted to 7.6 with solid potassium dihydrogen phosphate. This solution was then added to an ice-cold, stirred solution of enol phosphate (2.00 g, 3.45 mmol) in THF (25 mL). After 30 minutes, the reaction mixture was transferred to a pressure flask containing ether (40 mL) and 10% palladium on charcoal (2.0 g). This mixture was hydrogenated (310 kPa) for 1.25 hours. The reaction mixture was then diluted with ether (25 mL) and filtered.

117 7 8 0 9 4

The organic phase was separated and washed with water (2 x 5 mL). the combined aqueous phases were washed with ether (3 x 25 mL) and then concentrated in vacuo. Column chromatography (reverse phase, 45 x 130 mm, water as eluent), followed by freeze-drying of the carbapenem-containing fractions, gave 650 mg of crude product. This was rechromatographed to give pure title product (450 mg, 39%).

1 HNMR (D 2 O) δ: 1.24 (3H, d, J = 6.4 Hz), 3.19 (2H, q, 10 J = 2.6, 9.2 Hz), 3.45 (1H, q , J = 2.8, 6.0 Hz), 3.91 (3H, s), 4.06 (3H, s), 4.08-4.36 (2H, m), 4.54 (2H, d, J = 2.8 Hz), 8.71 (1H, s); IR (nujol phase) 1755 cm UV (phosphate buffer, pH 7.4) λ max: 294 nm (ε = 8.202); 15 Tl / 2 f phosphate buffer, pH 7.4, M = 0.067, T = 37 ° C), 9.1 hours.

Example 20 (1'R, 5R, 6S) -N- (1,3-dimethyl-5-tetrazolium) -methylthio [6- (1-hydroxyethyl) -7-oxo-1-azab-20-cyclo] 3,2,07hept-2-ene-2-karboksylaatti_

OH CH

25 \ o © A. 5-carbethoxy-2-methyltetrazole and 5-carbethoxy-1-methyltetrazole__ 30 N - N N ^ Nv N-e * Nv λ

Ϊ V ",» -full * | K + I

"'" C,' »,> Λ 0Et J" »W

35 3 118 78094 la. Methylation with diazomethane

A solution of 5-carbethoxytetrazole (9.17 g, 0.064 mmol) in ethyl ether (80 mL) was cooled to 0 ° C and treated by the dropwise addition of a solution of dazomethane (5 min, 15 min). 3 g, 0.071 mol) in ether (200 ml). The pale yellow solution was stirred for 30 minutes and the excess diazomethane was destroyed by the addition of acetic acid (1 mL). Evaporation of the solvent and distillation of the residue gave a clear oil: 10 kp. 95-100 ° C / 0.5 torr; 9.64 g (96%). 1 Hmr showed it to be a 6: 4 mixture of 1-methyl and 2-methyl isomers. Separation of the two isomers could not be performed by distillation or by hplc: ir (membrane) \) max: 1740 cm-· * · (ester C = 0); 15 Hmr (CDCl 3) 1.53 (3H, two overlapping t, J = 7.0, C 1 H 2 Cl 2), 4.46 and 4.53 (3H, 2S, 1-methyl and 2- CH 2 of methyltetrazoles, ratio 6: 4, methyl of the 2-isomer is in the lower field and is a minor part of the product), 4.5 ppm (2H, two overlapping q, C 2 CH 2) · 20 Ib. 5-carbethoxy-2-methyltetrazole CH3I f COjEi || Y_CO2EC + j ^ -COjEt 1J0.c 3 25 XCH3 CH3

A mixture of 5-carbethoxy-2-methyltetrazole and 5-carbethoxy-1-methyltetrazole (0.252 g, 1.61 mmol; the ratio of the two isomers was 1: 1) in iodomethane (0.5 mL) was sealed in a glass tube. and heated at 100 ° C for 15 hours and at 130 ° C for 6 hours. Distillation of the reaction mixture gave the title compound as a pale yellow oil: 0.139 g (55%); tr. 95-100 ° C / 0.5 torr (air bath temperature): 35 ^ D. Moderhack, Chem. Ber., 108, 887 (1975).

2 Using a mixture of ethanol and ether as the ratio of isomers, the same was obtained.

I; 78094 119 and (film) 0: 1740 cm -1 (ester C = O);

»ItldK, S I

XHmr (CDCl 3) δ: 1.46 (3H, t, J = 7.0, CH 3 CH 2), 4.53 (3H, s, CH 3-2), 4.5 (2H, q, J = 7.0, CH2CH3).

2. Methylation of dimethyl sulfate A solution of 5-carbethoxytetrazole (1.42 g, 0.01 mol) in dry acetone (20 ml) was treated with dry potassium carbonate (1.38 g, 0.01 mol) and dimethyl sulfate (1.26 mol). g, 0.01 mol). The mixture was heated at reflux for 12 hours. The carbonate was filtered off and the solvent was evaporated in vacuo. The residue was diluted with dichloromethane (30 ml), washed with saturated sodium hydrogen carbonate (10 ml), brine (10 ml) and dried over anhydrous sodium sulfate. Evaporation of the solvent and distillation in vacuo gave 15 clear oils: 1.45 g (93%); tr. 85-110 ° C / 0.5 torr.

^ Hmr showed two isomers in a 1: 1 ratio.

B. 5-Hydroxy imitated 1-2-methyl 3 tretazi + ubh <* £ / 1 1. By reduction of the mixture of esters 25 A mixture of 5-carbethoxy-1-methyltetrazole and 5-carbethoxy-2-methyltetrazole (6: 4) (7.60 g, 0.049 mol) in dry tetrahydrofuran (50 mL), cooled to 0 ° C and treated with lithium borohydride (1.06 g, 0.49 mol). mmol), which was added in small portions over 15 minutes. The mixture was kept at 10 ° C for another 30 minutes and then stirred at 20 ° C for 4 hours. The mixture was cooled to * 0 ° C and the excess hydride was carefully destroyed by the addition of 6-norm. HCl (pH 7 after no more gas evolved). The solution was concentrated in vacuo and the remaining 120 7 8 0 9 4 oil was diluted with dichloromethane (200 mL), washed with brine (10 mL) and finally dried over Na 2 O 4. Evaporation of the solvent and distillation of the residue in vacuo gave 1.83 g (33%) of a clear oil. The 5 Hmr of this material showed the product to be 5-hydroxymethyl-2-methyltetrazole.

2. By reduction of 5-carbethoxy-2-methyltetrazole

To a solution of 5-carbethoxy-2-methyltetrazole (0.139 g, 0.89 mmol, obtained by isomerization of a mixture of esters with methyl iodide) in dry tetrahydrofuran (1 mL) at 10 ° C was added solid lithium borohydride. (0.019 g, 0.87 mmol). The mixture was slowly warmed to room temperature and stirred for 4 hours. Excess borohydride was destroyed by the careful addition of -15 ti 6-norm. HCl at 0 ° C (pH 7). The solvent was evaporated and the residue was dissolved in dichloromethane (25 ml) and dried over anhydrous sodium sulfate. Evaporation of the solvent gave the title compound as a clear oil: 0.092 g (91%); tr. 90-120 ° C / 0.5 torr, with decomposition; ir (film) λ: 3350 cm -1 (broad, OH); 1 Hmr (CDCl 3) δ: 4.4 (2H, s, CH.j-2), 4.93 (2H, s, CH 2 O).

C. 5-Acetylmercaptomethyl-2-methyltetrazole 25

N X) hlSC1 'Et3N K ^ N

To a solution of 5-hydroxymethyl-2-methyltetrazole (1.83 g, 11.7 mmol) in dry dichloromethane (25 mL) at 0 ° C was added methanesulfonyl. chloride (1.47 g, 12.9 mmol), followed by the dropwise addition of triethylamine 35 (1.30 g, 12.9 mmol) over 5 minutes. The mixture was stirred at 0 ° C for

II

121,78094 n and then treated with a solution of potassium thioacetate (1.60 g, 14.0 mmol) in dry N, N-dimethylformamide (10 mL). The resulting gel was stirred at 0 ° C for 3 hours. The reaction mixture was diluted with dichloromethane (200 mL), washed with brine (20 mL) and dried over anhydrous sodium sulfate. Evaporation of the solvent in vacuo and chromatography of the resulting oil on silica gel (2 x 15 cm, eluting with dichloromethane and dichloromethane with 5% acetone) gave the title compound as a clear oil: 1.31 g (65%); and (film) max: 1696 cm -1 (thioester C = O); 1 Hmr (CDCl 3: 2.43 (3H, s, SAc), 4.36 (3H, s, 2-CH 3), 4.38 ppm (2H, s, 5-CH 2).

15 D. 5-Mercaptomethyl-1,3-dimethyltetrazolium trifluoromethanesulfonate_

CH

20 L ^ CB2 3 - - *

A solution of 5-acetylmercaptomethyl-2-methyltetrazole (0.400 g, 2.32 mmol) in dry di-. . In chloromethane (3 mL), treated with methyl triflate (0.76 g, 4.64 mmol) and stirred at 22 ° C for 16 h, evaporating the solvent in vacuo to give a red oil. This salt was dissolved in cold anhydrous water (5 mL) and treated with 4 mol. with sodium hydroxy-30 din (0.8 mL, 3.2 mmol). The mixture was stirred at 0 ° C for 40 minutes, diluted with water (7 mL), and the pH was adjusted to 7.3 with saturated K 2 PO 4. The resulting clear solution was kept under nitrogen and used immediately in the next step.

; ·. 35,122,78094 E. (1 ', R, 5R, 6S) -3- (1,3-dimethyl-5-tetrazolium) methylthio-6- (1-hydroxyethyl) -7-oxo-1-azabicyclo [3 , 2,07hept-2-ene-2-carboxylate 5 7 OH r ΛΧΟ- ^ '

CO PNB

CF SO® 3 3

20 OH

1 CH

h2p <2 / c v — rv -vTT 3

_i_>. I Vsch2V +) I

O. | 1 © CH, CO 2 3 25

A solution of enol phosphate (0.915 g, 1.58 mmol) in tetrahydrofuran (8 mL) was cooled to 0 ° C and treated by the dropwise addition of a solution of 5-mercaptomethyl-1,3-di-30-methyltetrazolium over 20 minutes. trifluoromethanesulfonate (2.32 mmol, prepared above). The pH of the reaction mixture remained stable at 6.5 throughout the addition. Another 20 mi-. . After 1 minute, the pH of the solution was adjusted to 7.0 with saturated sodium bicarbonate. The mixture was transferred to a hydrogenation flask, diluted with THF (10 mL), ether with 123,76094 (20 mL) and ice (20 g). Carbapenem was hydrogenated with 10% palladium on activated carbon at 45 psi, slowly raising the temperature to 22 ° C for 90 minutes. The catalyst was filtered off and washed with cold water (5 ml) and ether (20 ml). The aqueous phase was washed with ether (20 ml) and kept under vacuum for 20 minutes to remove residual organic solvent. Chromatography on PrePak 500-C / 18 and eluting with water gave the title compound as a white powder after freeze-drying 0.266 g (49%); + 13 ° (c 1.04, H 2 O); UV (H 2 O, pH 7.4) λ max: 294 nm (ε = 7,500); ir (KBr)> Jma · 1755-lactam C = 0), 1600 cm -1 (broad, carboxylate C = 0); 1 Hmr (D 2 O) δ: 1.24 (3H, d, J = 6.4 Hz, CH 3 CHOH), 3.0-3.3 (2H, m, H-4), 3.42 (1H, dd, J = 5.8, J = 2.9, H-6), 4-4.2 (2H, m, H-5 and CH 3 CHOH), 4.34 and 4.57 (2 x 3H, 2S, tetrazole CH 3 -1 and 3), 4.49 and 4.51 (2H, 2s, CH 2 S). The half-life of the product was 10.5 hours at 37 ° C (c 10 ~ 4 mol in pH 7.4 phosphate buffer).

20 Example 21

Alternative Method 3- (N-Methylpyridin-2-yl-methanethio) -6 * - [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.07] hept-2-ene-2-carboxy - to make a slab__ 25 ci_ 11 "CO © ®CHj 30 A.

To a 2 liter flask equipped with a magnetic stirrer and for distillation on a Vigreaux column, heating mantle and N 2, 4.0 moles (432 mL) of methyl acetoacetate and 8.0 moles (464.6 g of allyl-5 alcohol. The reaction mixture was distilled for 12 hours at 92 ° C.

136 ml (2.0 moles) of allyl alcohol were added and the mixture was distilled for 23 hours. 136 ml (2.0 moles) of allyl alcohol were then added thereto, and the mixture was distilled for 16 hours. The reaction mixture was then distilled in vacuo and the product 10 was collected at 105-110 ° C under a vacuum of 35 mm Hg.

414 g of allyl acetoacetate were obtained (yield 73%).

B.

0 ^ TsN, 9 15

To a solution of allylacetoacetate (226.5 g, 1.594 mol) in 3 liters of acetonitrile and triethylamine (243.4 mL, 1.753 mol) was added p-toluenesulfonyl azide (345.3 mL, 1.753 mol) over 1 hour while maintaining the temperature in a cooling bath. at 20 ° C. The reaction mixture turned yellow. The reaction mixture was then stirred at room temperature under nitrogen for 18 hours. The mixture was concentrated on a rotary evaporator. The residue was dissolved in diethyl ether (2.6 L) and 1 mol. To an aqueous solution of KOH (800 mL). The organic phase was washed five times with 1 mol. With KOH (500 mL) and once with brine (400 mL). After drying over MgSO 4 and concentration on a rotary evaporator (temp. <30 ° C), 260.2 g (97%) of the title product are obtained.

C.

0 ^ _ osi4- 35 2 -> N2 I: 125 7 80 9 4

To a stirred suspension of allyl diazoacetate (203 g, 1.195 moles) in 2 liters of methylene chloride and 199 mL (1.434 moles) of triethylamine at 5 ° C was added 302 mL (1.315 moles) of 5-t-butyldimethylsilyl triflate over 45 minutes. . The mixture was stirred for 1 hour at 5 ° C and then for another hour without cooling. The reaction mixture was washed 4 times with 500 mL of IVO and then once with 500 mL of brine. It was then dried over Na 2 SO 4 and concentrated to 344 grams of an orange oil. This oil was used as such in the next step.

D.

15 osi-f ^ OCOCH, CT H ZnCl2 ^ 20 I, OSi-f N, 25

To a mixture of (1'R, 3R, 4R) -3- (1'-tert-butyldimethylsilyloxyethyl) -4-acetoxyazetidin-2-one (28.7 g, 0.1 mol) and freshly melted ZnCl 2 (6.8 g, 0.05 mol) in dry CH 2 Cl 2 (700 mL) was added dropwise over 5 hours to a solution of allyl-2-diazo-3-tert-butyldimethylsilyloxy-3 butenoate (33.84 g, 0.12 mol) in CH 2 Cl 2 (50 mL). The mixture was stirred at room temperature for 2 hours during which time TLC (thin layer chromatographic analysis) indicated the small amount of starting material remaining. An additional portion of allyl 2-diazo-3-tert-butyl 126,78,09 4 dimethylsilyloxy-3-butenoate (4.23 g, 0.015 mol) in 10 mL of Cl 2 Cl 2 was added over 1 hour and stirring was continued at room temperature. at 10 hours. The reaction mixture was then diluted with ethyl acetate (750 mL), washed (2 x 300 with 5 mL saturated NaHCO 3, 300 mL brine), dried (MgSO 4) and evaporated to dryness to give 62.5 g of a dark orange oil which was dissolved in methanol (500 mL) and treated with 1N aqueous HCl (110 mL). The resulting mixture was stirred at 10 room temperature for 2 hours, after which time 10 ml of 1-norm was added. HCl and then stirred for an additional 2 hours. The reaction mixture was concentrated to half its volume and poured into a mixture of ethyl acetate (800 ml) and water (800 ml). The organic phase was separated, washed with water (800 ml) and the combined aqueous extracts washed with ethyl acetate (400 ml). The combined organic extracts were washed with brine (2 x 400 mL), dried (MgSO 4) and concentrated to a 32 g dark orange-red oil. Chromatography ("flash") gave 9.33 g (33% yield) of the title product as a golden yellow oil which solidified to a pale yellow solid.

1 H-nmr (CDCl 3) δ: 6.20-5.72 (m, 2H), 5.48-5.21 (m, 2H), 4.74 (dt, J = 5.8, J '= 1 , 2 Hz, 2H), 4.30-3.88 (m, 2H), 3.30-3.20 (m, 2H), 2.89 (dd, J = 7.3, J '= 2 (1H, 1H), 2.18 (s, 1H), 1.32 (d, J = 6.2, 3H).

E.

OSi-f- N2; "*

OH

; 35 127 7 8 0 9 4

A mixture of the oiaziazo ester prepared in Step D above (9.2 g, 32.7 mmol) and rhodium acetate ^ Rh 2 (OAc) 47 in benzene (1 L) was boiled for 1 h. The solution was treated with activated carbon 5 and filtered through a pad of Celite. The mattress was washed with 100 ml of hot benzene. Concentration of the filtrate gave 8.08 g (97% yield) of the title product as a light brown crystalline solid.

1 H-nmr (CDCl 3) 6.15-5.68 (m, 1H), 5.45-5.18 (m, 2H), 4.71-4.60 (m, 2H), 4.40- 4.05 (m, 2H), 3.17 (dd, J = 7.1, J1 = 2.0, 1H), 2.95 (dd, J = 6.9, J '= 18.9, 1H ), 2.42 (dd, J = 7.6, J '= 18.8, 1H), 1.88 (s, 1H), 1.39 (d, J = 6.3, 3H).

F.

OH

“^ Yct, -►

OH

To a solution of the ketoester from Step E (7.5 g, 0.03 mol) was added diisopropylamine (6.08 mL, 0.035 mol) at 0 ° C under atmospheric protection followed by diphenylphosphoryl chloride. After 15 minutes, TLC analysis indicated that no starting material remained. To the reaction mixture was added di-isopropylamine (6.26 mL, 0.036 mol) and a solution of freshly distilled 2-mercaptomethylpyridine (4.5 g, 0.036 mol) in 5 mL of acetonitrile. After stirring for 2 h at 0 ° C, the mixture was poured into ethyl acetate (1 L), washed with water (2 x 150 mL), saturated NaHCO 3 (150 mL), H 2 O (150 mL), and brine. la (200 ml). The organic phase was dried (MgSO 4) and concentrated to a dark orange-yellow resin. Chromatography ("flash") gave the product as a golden yellow oil. The product was dissolved in diethyl ether and cooled to 0 ° C. Filtration gave 4.8 g (yield to 44%) of the purified title product as cream crystals.

1 H-nmr (CDCl 3) δ: 8.6-8.4 (m, 1H), 7.85-7.15 (m, 3H), 6.20-5.74 (m, 1H), δ , 54-5.15 (m, 2H), 4.80-4.66 (m, 2H), 4.29-4.03 (m, 1H), 4.19 (s, 2H), 3, 69-2.85 (m, 1 H), 2.97 (s, 1 H), 1.32 (d, J = 6.2, 3 H).

G.

To a solution of the α-25 yl ester prepared in Step F (1.79 g, 4.97 mmol), tetrakistriphenylphosphine palladium (175 mg, 0.15 mmol) and triphenylphosphine (175 mg, 0.67 mmol) In CH 2 Cl 2 (25 mL), a solution of potassium 2-ethylhexanoate (1.085 g, 5.96 mmol) in ethyl acetate (12 mL) was added.

After stirring for 30 hours at room temperature, TLC analysis showed only traces of starting material. The reaction mixture was diluted with anhydrous diethyl ether (150 mL) and the precipitate was collected by filtration, washed with ethyl acetate and then ether to give a light brown solid. This solid was dissolved in H 2 O (10 mL) and purified by reverse phase chromatography to give 1.85 g of the title product as a cream solid. This material was further purified by slurrying in acetone to give 1.47 g (83%) of pure title product.

1 H-nmr (D 2 O) δ: 8.45-8.36 (in, 1H), 7.92-7.22 (m, 3H), 4.78-3.91 (m, 2H), 4.69 (s, 2H), 3.34-2.71 (m, 3H), 1.19 (d, J = 6.4, 3H).

B.

10

15 ° X

Toluenesulfonic acid (27.6 mg, 0.16 mmol) was added to a cooled (0 ° C) suspension of potassium 6-hydroxyethyl-2- (2-pyridylmethylthio) carbapenem. -3-carboxylate (53.8 mg, 0.15 mmol) in acetone (2 mL). The mixture was stirred at 0 ° C for 20 minutes and then treated with methyl triflate (0.02 mL). After stirring for 60 minutes at 0 ° C, LA-1 resin was added followed by hexane (6 mL). The mixture was extracted with water (4 x 0.5 mL) and the combined aqueous phases were purified by reverse phase HPLC to give 10 mg of the title product.

130 78094

Example 22 3- (N-Methylpyridin-2-yl-methanethio) -6H- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.2] hept-2-ene-2 preparation of carboxylate by pot "method" ohh h • tQyig) 10 2 OH CF-SO © T CH7 3 3 Λγ-τΛ © t ^ c'o2pnb 3

20 O

CH 3 CN ClUlCO 3 NaOH

0-0.45 min. EtN (iPr) 2 8.0, 0 · 0. 1 h.

25 and J,

OH

^ \ 0 ΪΗ3 CF SO ©. s;:? cr ·· 2 / pH 6.5-7.5 THF, Ho0 35 Ψ 2 131 78094 Λτ-κ.

5 CO2PNB

5 η2 pd-c ιο% 5eC, 2 h.

10 *

OH

T CH, 15 £ 2 A. Preparation of enol phosphate (2)

20 SH OH

X H H y H H O

Ji> -► J1> 0P <W2

0X '5. 0X

C02PMB co2pnb 25 1 2 An ice-cold solution of ketone 1 (3 g, 8.62 30 mmol) in acetonitrile (30 mL) was treated with ethyldiisopropylamine (9 mmol, 1.04 eq., 1.57 mL) (addition time ca. 2 minutes) and chlorodiphenyl phosphate (9 mmol, 1.04 equiv, 1.87 mL) (addition time about 2 minutes). The reaction mixture was stirred for 45 minutes to 35 minutes and TLC analysis (ethyl acetate, silica gel) showed that ketone 1 disappeared. The solution was diluted with ethyl acetate (60 mL), washed with cold water 132.

78094 (2 χ 50 mL) and brine, dried over sodium sulfate and concentrated (bath temperature below 20 ° C) to give a foam which was used as such.

5 B. Preparation of thiol (4)

Tl © © T3 ^ 11 NaOH / H2 ° „A e 'Ha“ 2) KH2PO4 2 4 15 An ice-cooled solution of room acetate 3 (3.31 g, 10 mmol) in water, which had been purged with nitrogen for 5 minutes , was treated by dropwise addition (over about 5 minutes) of a cooled solution of sodium hydroxide (1.75 equiv., 17.5 mmol-20 l, 0.7 g) in water (8 ml). The mixture turned yellow.

After 75 minutes under a nitrogen purge, the pH was adjusted to 7.4 with saturated aqueous Itt ^ PO ^ sn. The reaction mixture was diluted with water (15 mL).

This aqueous solution of thiol 4 (50 mL, 0.2 mmol / mL) was used as such.

133 78094 C. Wiring

5 0H A

^ rrvJw> 2 * LL / 5 pH 6'5-7'.5 °> o2pnb ° ·· 1 »·

2 £ H} β CH

10 "X © f3

C02PNB

An ice-cooled solution of compound 2 (crude, prepared at A, 8.62 mmol) in tetrahydrofuran (50 mL) was treated by the dropwise addition of an aqueous solution of thiol 4 prepared in B (5 mL of a solution every 5 minutes). During the reaction, the pH of the reaction mixture was maintained between about 6.5 and 7.5 (primarily at u) by the addition of cooled 2-norm. natriumhydrok-hydroxide solution. The reaction was monitored by TLC using (a) silica gel, ethyl acetate; (b) reverse phase Analtech-RPSF technique, CH 2 CN-pH 7 buffer (4: 6).

Finally, 1.15 equiv. thiol (50 ml of solution). The reaction was complete after 1 hour at 0 ° C and the mixture was used as such for hydrogenation after adjusting the pH to 7.

30 134 78094 D. Hydrogen §H J3 CH 310 5 A ^ ^ ^ ^]] H2 / Pd-C10 * C-) THF-H2O-ether ^ CO, PNB 0-C, 2 h.

- OH CH

10> Λ>. ^ © £ 3 6 15

The reaction mixture containing Compound 5 (prepared in C) was transferred to a Parr flask containing THF (10 mL), phosphate buffer (pH 7, 0.1 M) (10 mL), ether (75 mL) and 10% Pd. With -C catalyst (5 g) and hydrogenated at 310 kPa at 3-10 ° C for 2 hours. The catalyst was then filtered off, washed with water (3 x 10 ml) and the pH was carefully adjusted to 6.2 with cold 2-norm. Na0H, respectively. Ether was added and the aqueous phase was separated and washed again with ether. The aqueous phase was rinsed in vacuo free of organic solvent and then purified on a Bondapak-C-18 column (100 g, 4.5 x 13 cm) with cold distilled water. The light yellow fractions containing the product (checked by UV and TLC) were lyophilized to give 1.46 g of compound (50%) * as a yellow powder. Jk.293, δ = 9000, λ 271, £ = 11064.

35 * yield calculated from bicyclic ketone.

135 78094

Example 23 (5R, 6S) -3 - [(11,3-dimethylpyridinium-4-yl) methyl] thio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2.0] hept- 2-ene-2-carboxylate (23Ά) 5 j.

(4R, 5R, 6S) -3- [E (1,3-dimethylpyridinium-4-yl) methyl] -7-thio} -6- [1- (1H) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo Preparation of / / 3.2.Q7hept-2-ene-2-carboxylate (23B) 10 Γ 5 fH3

A'j-A

'Co | N: h3 15

23A R = H

23B R = CH3 A. Preparation of 4-hydroxymethyl-3-methylpyridine

CH-OH

CH, 2 (& "* -8¾—> (o / '25 ^ ir e 25 3) H30® The general method of Boekelheiden ^ was used to prepare hydroxymethylpyridines. Thus, a freshly distilled solution of 3,4-lutidine (46.0 g, 0.34 mol) in 120 ml of glacial acetic acid 30 was cooled to 0 [deg.] C. and then 64 ml of 30% H2O2 were added dropwise, and the resulting solution was heated at 75 [deg.] C. (oil bath temperature) for 3 hours. 30% H 2 O 2 was added and heated for 18 h.Finally, 20 mL of 30% H 2 O 2 was added again and the reaction was maintained at 75 ° C for another three hours.The solution was then concentrated to about 100 mL with a water jet. , 50 mL of H 2 O was added and the mixture was concentrated to about half volume, the resulting mixture was cooled (0-5 ° C) and basified, pH about 10, with cold 40% aqueous NaOH, then extracted with CH 2 Cl 2. (5x) and the extract was dried (Na 2 CO 3 + Na 2 SO 4) and concentrated by rotary vacuum to give a yellow solution 5. Dilute this solution with hexane. , a solid was collected which was collected by filtration and then dried in vacuo to give 3,4-lutidine N-oxide (48.0 g, 83%) as an off-white solid.

The N-oxide was gradually added to 60 ml of acetic anhydride, and the resulting dark orange solution was heated (water bath) at about 90 ° C for 1 hour. Excess acetic anhydride was then distilled off under reduced pressure and the material boiling at 90-120 ° C / 13 Pass (39.0 g) was collected. Chromatography of this oil (silica gel / ethyl acetate-pet.

Ether = 2: 3) gave pure 4-acetoxymethyl-3-methylpyridine (19.0 g, 30%) as an oil: infrared (undiluted 1745 cm 1).

The acetate and 100 mL of 10% aqueous HCl were then refluxed for 1 hour. The resulting solution was cooled to 0 ° C, basified with solid CH 2 Cl 2 and then extracted with CH 2 Cl 2 (3 x 100 mL). The organic extract was washed (brine), dried (Na 2 SO 4) and evaporated to give 11.0 of an off-white solid, mp 70-72 ° C. This solid was triturated with dry cold ether to give pure 4-hydroxymethyl-3-methylpyridine (9.5 g, 67%) as a white solid, mp 77-80 ° C (m.p. 81-82 ° C). C): 1 Hnmr (CDCl 3) 8.27, 7.41 (ABq, J = 5Hz, 2H), 8.18 (s, 1H), 5.63 (br s, -OH), 4.67 (s, CH2), 2.20 (s, CH3); infrared (nujol) 3170 cm "1.

30 1. V. Boekelheide, W.J. Linn, JACS, 76, 1286 (1954).

2. W.L.F. Armarego, B.A. Milloy, S.C. Sharma, JCS, 2485 (1972).

78094 137 B. Preparation of 4- (acetylthiomethyl) -3-methylpyridine

CHo0H _A

I 2 ch2sac • -> dr To an ice-cold, mechanically stirred solution of triphenylphosphine (17.04 g, 0.065 mol) in 250 mL of THF was added dropwise diisopropyl azodicarboxylate (12.8 mL, 0.065 mol), and the resulting suspension was stirred at 0 ° C for 1 hour. To this mixture was added dropwise a solution of 4-hydroxymethyl-3-methylpyridine (4.0 g, 0.0325 mol) in 100 mL of dry THF, followed by freshly distilled thioacetic acid (4.64 mL, 0.065 mol). ). The resulting mixture was stirred at 0 ° C for 1 hour to give an orange solution. The solution was concentrated (rotary evaporator) and then diluted with petroleum ether. The resulting mixture was filtered and the filtrate evaporated to give an orange 2q oil. This oil was chromatographed (silica gel / hexane, then 10% -50% ethyl acetate-hexane) to give pure product (6.0 g, 100%) as a yellow oil, m.p. (air bath temperature) 95-100 ° C / 13 Pa 1Hnmr (CDCl 3) 8.40, 7.20 (ABq, J = 5Hz, 2H), 8.37 (s, 1H), 4.08 (s, CH 2) , 2.35 (s, CH3), 2.32 (s, CH3); infrared (neat) 1695 cm -1 C. Preparation of 4- (acetylthiomethyl) -1,3-trimethylpyridinium triflate CH ~ SAc CHpSAc j 2 30 ch3 XXCH3 oi -> I oj θ CF3S ° 3 ch3 35 138 78094 To an ice-cold solution containing thioacetate (2.95 g, 0.016 mol) in 10 ml of methylene chloride, trifluoromethanesulfonate (4.60 ml, 0.04 mol) was added dropwise and the mixture was stirred at 0 ° C in the presence of nitrogen for one hour.

The reaction mixture was then evaporated to dryness and the residue triturated dry with ether. The resulting solid was collected by filtration and dried in vacuo to give the product (4.0 g, 72%) as a white solid: 1 Hnmr (CDCl 3) δ · 8.72 (s, 1H), 8.58, 7.87 (ABq , J = 6Hz, 2H), 4.39 (s, N-CH 3), 4.17 (s, CH 2), 2.53 (s, CH 3), 2.36 (s, CH 2); infrared (pure) 1700 cm D. (5R, 6S) -3 - [[(1,3-dimethylpyridinium-4-yl) methyl] thio] -3- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo -1 5 / 3.2.0 ._ / Preparation of hept-2-ene-2-carboxylate.

CHjSAe OK M · 20 O rP1 — ► ®L “i® CT SO® °“ V ™

3) H2. Pd / C

To an ice-cold, N 2 O-treated solution of NaOH (0.324 g, 0.008 mol) in 10 mL of H 2 O was added thioacetate (1.40 g, 0.004 mol) and the mixture was stirred at 0 ° C in the presence of nitrogen. for one hour. After adjusting the pH to 7.2-7.3 using 3% potassium dihydrogen phosphate, the resulting solution was added dropwise to an ice-cold solution of enol phosphate (1.45 g, 0.0025 mol) in 20 mL of THF. The mixture was stirred at 0 ° C for 1 hour and then transferred to a pressure flask. To this mixture were added 20 ml of ether, 25 ml of 0.1 M phosphate buffer solution (pH 7.4) and 1.4 g of 10% palladium on carbon. The mixture was then hydrogenated at 310 kPa for one hour. The reaction mixture was filtered through a diatomaceous earth filter, 78094 139 and the filter was washed with ether and phosphate buffer solution, pH 7.4. The aqueous phase was separated and the remaining solvents were removed in vacuo. The resulting aqueous solution was applied to a phase transfer column (C1t> BondaPak) eluting with 10 5 H 2 O and then 10% acetonitrile-H 2 O. Lyophilization of the appropriate fractions gave 0.9 g of an orange solid.% This material was rechromatographed using H 2 O and then 2% acetonitrile-H 2 O as eluent. Lyophilization gave pure 23A (0.25 g, 57%) as a yellow solid: 10 1Hnmr (ϋ20) <5 · 8.55 (s, 1H), 8.53, 7.96 (ABq, J = 6.8 Hz). , 2H), 4.30-3.99 (m, 2H), 4.27 (s, 5H), 3.35 (dd, J1 = 2.8 Hz, J2 = 6.0

Hz, 1H), 3.05 (d, J = 8.8 Hz, 2H), 2.50 (s, 3H), 1.23 (d, J = 6.3 _ 1

Hz, 3H); infrared (KBr) 1755, 1590 cm; uv (phosphate type solution, pH 7) 295 nm (7180).

E. (4R, 5R, 6S) -3- [Z- (1,3-dimethylpyridinium-4-yl) methyl] thio] -6- (R) -hydroxyethyl] -4-methyl-7-oxo- 1-azabicycloZ ~ 3.2. Preparation of Q7hept-2-ene-2-carboxylate.

20 “j ™ T © M ·

3) Hj. Pd / C

An ice-cold solution of the bicyclic ketone (0.906 g, 0.0025 mol) in 10 mL of acetonitrile was treated sequentially with diphenyl chlorophosphate (0.544 mL, 0.00263 mol), diisopropylethylamine (0.457 mL, 0.00263 mol) and 4-dimethyl aminopyridine (0.3 mg). After 50 minutes, the reaction mixture was diluted with cold ethyl acetate and then washed with cold water and salted. The organic phase was dried (Na 2 SO 4) and evaporated at room temperature to give enol phosphate as an off-white foam. This foam was treated with 20 mL of THF, cooled to -30 ° C in the presence of nitrogen, and then treated with aqueous thiolate / prepared as before from 0.3 g of NaOH 5 (7.5 mmol) and 1.3 g of g of thioacetate (3.76 mmol) in 10 ml of IVO. The reaction mixture was stirred at -30 ° C for 30 minutes, then at 0 ° C for 75 minutes and finally transferred to a pressure flask containing 20 ml of ether, 30 ml of 0.1 M phosphate buffer solution (pH 7.4) and 1.5 g of 10% palladium-on-carbon. The mixture was hydrogenated at 310 kPa for 1 hour, then filtered through diatomaceous earth and the aqueous phase separated and concentrated in vacuo. The resulting solution was applied to a phase transfer column (CH 2 BondaPak) eluting with H 2 O. Lyophilization of the appropriate fractions gave 1.2 g of a yellow solid. This material was rechromatographed (eluting with H 2 O and 4% acetonitrile-H 2 O) to give, after lyophilization, pure 23B (0.250 g, 28%) as a yellow solid: Hnmr (D 2 O) <18.53 ( s, 1H), 8.49, 7.81 (ABq, J = 6.2 Hz, 2H), 4.38-3.98 (m, 4H), 4.27 (s, 3H), 3.49. -20 3.18 (m, 2H), 2.51 (s, 3H), 1.25 (d, J = 6.7 Hz, 3H), 1.16 (d, J = 7.6 Hz, 3H ); infrared (KBr) 1750, 1595 cm-2; uv (phosphate buffer solution, pH 7) 292 nm (£ 7930).

Example 24 (5R, 6S) -3 - ((Cl, 1,2-dimethylpyridinium-4-yl) methyl] thio-6- (Z-1- (R) -hydroxyethyl) -7-oxo-1-azabicyclo [5.2 . Preparation of Oj-hept-2-ene-2-carboxylate (28A) 30 and (4R, 5R, 6S) -3- (2-dimethylpyridinium-4-yl) methylthio] -6- [1- (R) -hydroxyethyl] Preparation of -4-methyl-7-oxo-1-azabicyclo [3.3.0] hept-2-ene-2-carboxylate (28B) i 35 141 78094

OH R

X Λ CH0 JCC / s / YoT "_ le 5 CO® ® CH3

28A R = H

28B R = CH3 10 A. Preparation of 4-hydroxymethyl-2-methylpyridine

„W CH-OH

C09Me | 2 15 (oi - »(¾

To a suspension of 95% lithium aluminum hydride 20 (2.4 g, 0.06 mol) in 150 mL of anhydrous ether was added a solution of 2-methyl isonicotinate (14.0 g, 0.093 mol) in 50 mL of anhydrous ether. At 5 ° C in the presence of nitrogen. The resulting mixture was stirred at room temperature for 30 minutes and then refluxed for 2 hours. An additional 1.2 g (0.03 mol) of lithium aluminum hydride was gradually added and refluxing was continued for one hour. The reaction mixture was then cooled to 0 ° C and treated sequentially with 3.75 mL of Orta, 3.75 mL of 15% aqueous NaOH, and finally 11.25 mL of H 2 O. This suspension was then filtered and the filter cake was washed with ether and then ethyl acetate. The filtrate was evaporated to give a dark yellow oil which was treated with acetonitrile and then filtered through a silica gel filter (eluting with acetonitrile and then acetone). This gave product 35 (7.7 g, 67%) as a yellow oil: 1 Hnmr (CDCl 3) δ 8.30, 7.10 142 7 8 0 9 4 (ABq, J = 5 Hz, 2H), 7.17 (s , 1H), 5.42 (s, -OH), 4.70 (s, CH 2), 2.50 (s, CH 3).

O. Efimovsky, P. Rumpf, Bull. soc. chim. Fr., 648 (1954).

5 B. Preparation of 4- (acetylthiomethyl) -2-methylpyridine CH OH CH2SAc "& -> (§,

To a solution of triphenylphosphine (31.4 g, 0.12 mol) in 200 mL of dry THF was added dropwise diisopropyl azodicarboxylate (23.6 mL) at -5 ° C in the presence of nitrogen. , 0.12 mol), and the mixture was stirred at -5 ° C for 1 hour. To the resulting suspension was added over about 10 minutes a solution of 4-hydroxymethyl-2-methylpyridine (7.60 g, 0.062 mol) and freshly distilled thioacetic acid (8.60 mL, 0.12 mol) in 40 mL of dry THF: a. The reaction was stirred at 0 ° C for 30 minutes and then at room temperature for 1 hour. The resulting suspension was filtered and the filtrate was concentrated to give an orange-yellow liquid which was diluted with ether and filtered. The filtrate was evaporated and the residual oil was chromatographed (silica gel / ethyl acetate-hexane = 1: 1) to give thioacetate (8.87 g, 79%) as a yellow oil: ΊΗηπΐΓ (CDCl 3) 8.45, 7.03 (ABq, J = 5 Hz, 2H), 7.08 (s, 1H), 4.04 (s, CH 2), 2.55 (s, CH),

Z -1 J

2.39 (s, CH 3); infrared (undiluted) 1695 cm 143 78094 C. Preparation of 4-mercapto-2-methylpyridine

CH, SAc CH, SH

5 (° X - »(° 1

To 15 mL of ice-cold, nitrogen-treated 1N NaOH was added 4- (acetylthiomethyl) -2-methylpyridine (1.358 g, 0.0075 mol) all in one portion. The reaction mixture was stirred for 15 minutes at 0 ° C, then washed with ether (2 x 5 mL), neutralized with concentrated HCl and extracted with methylene chloride (3 x 10 mL). The methylene chloride solution was evaporated to give the thiol (0.89 g, 96%) as a pale yellow oil which gradually turned pink: 1Hnmr (CDCl 3) δ 8.43, 7.37 (ABq, J = 5 Hz, 2H), 7 , 43 (s, 1H), 3.63 (d, J = 7.5 Hz, CH-), 2.55 (s, CH_), 1.81 (t, J = 7.5 Hz, 20 SH) .

D. Allyl- (5R, 6S) -3-H- (2-methylpyridin-4-yl) methyl] thio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo [3.2 Preparation of j0hept-2-ene-2-carboxylate.

25

OH OH

—- xJZ> ^ Ίoίc ',

Ho, I

CO 2 To an ice-cold solution of bicyclic ketone (0.760 g, 0.003 mol) in 8 mL of acetonitrile was added sequentially diphenyl chlorophosphate (0.653 mL, 0.00315 mol), diisopropyl-35-ethylamine (0.550 mL, 0.00315 mol), and dimethylaminopyridine (0.8 mg). After stirring at 0 ° C for 144 7 8 0 9 4, the mixture was cooled to -20 ° C with 4-mercaptomethyl-2-methylpyridine (0.620 g, 0.00446 mol) followed by di- isopropylethylamine (0.550 mL, 0.0035 mol) was added. The reaction was stirred at -20 ° C for 1.5 hours and then allowed to warm to room temperature. The resulting mixture was diluted with 50 mL of ethyl acetate, washed with CH 2 Cl 2, saturated NaHCO 3, saturated NH 4 Cl), dried (Na 2 SO 4) and evaporated. The residue was chromatographed on silica gel (eluted with ethyl acetate then acetonitrile) to give a solid which was triturated with dry ether to give pure product (0.820 g, 73%) as a white solid: 1 Hnmr (CDCl 3). Δ 8.45, 7.09 (ABq, J = 5 Hz, 2H), 7.15 (s, 1H), 6.25-5.80 (m, 1H), 5.60-5.20 (m, 2H), 4.82-4.68 (m, 2H), 4.55-4.05 (m, 2H), 3.97 (s, 2H), 3.16-2.93 ( m, 3H), 2.55 (s, 3H), 1.84 (br s, 1H), 1.32 (d, J = 6 Hz, 3H); infrared (undiluted) 1777, 1695 cm.

E. (5R, 6S) -3- (C [1,2-dimethylpyridinium-4-yl) methyl] -thio] -6- [1- (R) -hydroxyethyl] -7-oxo-1-azabicyclo / 3.2.07- preparation of hept-2-ene-2-carboxylate.

fiH OH

v & er -

A solution of the allyl ester (0.350 g, 0.936 mol) in 30 mL of 6 mL of dry acetonitrile was cooled to -5 ° C and treated with methyl trifluoromethanesulfonate (0.111 mL, 0.983 mol). After 15 minutes, a solution of tetrakis (triphenylphosphine) palladium (0.027 g, 2.5 mol%) and triphenylphosphine (0.027 g) was added. After stirring the reaction mixture for 5 minutes, pyrrolidine (0.082 mL, 0.983 mol) was added dropwise. The solid slowly began to separate from the resulting brown solution. The mixture was stirred vigorously at 0 ° C for 20 minutes, then 15 mL of cold (0 ° C) acetone was added slowly, and stirring was continued at 0 ° C for 20 minutes. The resulting suspension was filtered and the residue was washed with cold acetone and then dried in vacuo to give 0.345 g of a beige powder. This material was treated with a small amount of phosphate buffer solution (0.05M) at pH 7 and applied to a short phase transfer column (C 2 g BondaPak). Elution was performed with H 2 O and the relevant fractions were lyophilized to give 0.255 g of a pale yellow solid. This material was rechromatographed as before to give (after lyophilization) pure 24A (0.195 g, 60%) as a pale yellow solid: 1Hnmr (D 2 O) <f8.58, 7.83 (ABq, J = 6.4 Hz, 2H) 7.87 (s, 1H), 4.32-3.95 (m, 2H), 4.22 (s, 2H), 4.17 (s, 3H), 3.32 (dd, J1 = 2.6 Hz, J2 = 6.1 Hz, 1H), 3.06-2.93 (m, 2H), 2.74 (s, 3H), 1.22 (d, J = 6.4) Hz, 3H); infrared (KBr) 1757, 1590 cm -1 S (phosphate buffer, pH 7.4) 296 nm (¢ 7446).

F. F. Allyl- (4R, 5R, 6S) -3 - [(2-methylpyridin-4-yl) methyl] thio-3- [1- (R) -hydroxyethyl] -4-methyl-7-oxo-1 -azabicyclo / 3 ♦ 2.07hept-2-ene-2-carboxylate preparation.

25 oh λ r p _ - * Ή- ^ r · 30

A solution of the diazo ester (1.50 g, 0.00508 mol) in 12 mL of ethyl acetate-hexane (3: 1) was gently heated to reflux in the presence of nitrogen, and then 0.020 g of rhodium octanoate was added all in one 35 times. Rapid evolution of N 2 was observed for about five minutes, and after refluxing was continued for another 10 minutes, the reaction was complete (road). The solvents 78094 146 were later removed under reduced pressure and the remaining gum was treated with 15 ml of acetonitrile. The solution was cooled to -5 ° C and treated with diphenyl chlorophosphate (1.10 mL, 0.00533 mol), diisopropylethylamine 5 (0.927 mL, 0.00533 mol) and 4-dimethylaminopyridine (0.6 mg, 0, 1 mol%). The reaction mixture was stirred at 0 ° C for 1 h and then cooled to -20 ° C and first treated with a solution of 4-mercaptomethyl-2-methylpyridine (0.656 g, 0.00533 mol) in 1 mL of acetonitrile, then 0.927 with ml (0.00533 mol) of diisopropylethylamine.

The resulting mixture was stirred at -10 ° C for 1.5 h and then further treated with thiol (0.066 g, 0.53 mmol) and diisopropylethylamine (0.093 mL, 0.53 mmol). The reaction was allowed to warm to about 10 ° C for 1 h and then diluted with 75 mL of cold ethyl acetate, washed (H 2 O, brine), dried (Na 2 SO 4) and evaporated (bath temperature <30 ° C). The resulting gum was chromatographed on silica gel. Elution with ethyl acetate removed the impurities, and further elution with acetonitrile gave the product (1.04 g, 53%) as a pale yellow foam: 1 Hnmr (CDCl 3) 8.43, 7.07 / ABq, J = 5 Hz, 2H). 7.10 (s, 1H), 6.20-5.75 (m, 1H), 5.51-5.29 (m, 2H), 4.81-4.69 (m, 2H), 3, 96 (s, 2H), 3.35-3.05 (m, 2H), 2.53 (s, 3H), 2.16 (br s, 1H), 1.33 (d, J = 6.3) Hz, 3H), 1.22 (d, J = 7.3 Hz, 3H); infrared (undiluted) 1770, 1705 cm -1.

25 G. (4R, 5R, 6S) -3 - ((1,2,2-dimethylpyridinium-4-yl) methyl] thio] -3- [6- (R) -hydroxyethyl] -4-methyl-7-oxo-1 -azabicyclo / 3.2.Q7hept-2-ene-2-carboxylate ♦ 30 OH CHj OH qj CO2 ^ tO® 35 147 7 8 0 9 4

A solution of allyl ester (0.582 g, 0.0015 mol) in 15 mL of dry acetonitrile was treated with methyl trifluoromethanesulfonate (0.178 mL, 1.575 mmol) at -5 ° C in the presence of nitrogen. After 15 minutes, a solution of tetrakis (triphenylphosphine) palladium (0.035 g 2 mol%) and triphenylphosphine (0.035 g) in one ml of methylene chloride was added first, then after 5 minutes 0.131 ml (1.575 mmol) of pyrrolidine. The resulting mixture was stirred at 0 ° C for 20 minutes and then 30 ml of cold (0 ° C) acetone was added. The mixture was stirred vigorously at 0 ° C for 15 minutes, and the precipitate was collected by filtration, washed with cold acetone and dried in vacuo to give 0.520 g of a beige powder. Dilution of the filtrate with ether gave an additional 0.041 g of crude product. The combined solids were dissolved in a small amount of phosphate buffer (0.05H, pH 7.4) and applied to a phase transfer column (CH 2 BondaPak), eluting with CH 2 Cl 2 and then 2% acetonitrile. obtained, after lyophilization, 24B (0.413 g 76%) as a yellow solid: 1Hnmr (D2O) ^ 8.55, 7.76 (ABq, J = 6.3 Hz, 2H), 7.81 (s, 1H) , 4.4-3.7 (m, 2H), 4.19 (s, 2H), 4.16 (s, 3H), 3.47-4.14 (m, 2H), 2.73 (s, 3H), 1.24 (d, J = 6.4 Hz, 3H), 1.6 (d, J = 7.3 Hz, 3H), infrared (KBr) -1 1750, 1595 cm -1, phosphate buffer, pH 7.4) 293 nm (£ 7170).

25

Example 25 (5R, 6S) 3- (R, 6-dimethyl-pyridinium-2-yl) methylthio] -6- (1R-hydroxyethyl) -4R-methyl-7-oxo-1-azabicyclo / 3.2.07-30 hept Preparation of 2-ene-2-carboxylate COO Θ 78094 148 un A. (1,6-Dimethylpyridinium-2-yl) methylthiol, trifluoromethanesulfonate salt.

3 k3 10

A solution of (6-methylpyridin-2-yl) methyl thioacetate (1.0 g, 5.52 mmol) in dry ether (5 mL) maintained under a nitrogen atmosphere was treated with methyl triflate (0.74 mL, 6.5 mmol) and stirred at 23 ° C for four hours. The ether was decanted and the white solid was washed twice with ether (2 mL) and dissolved in hydrochloric acid (15 mL, 6N, 90.0 mmol). The resulting solution was heated at 70 ° C for four hours under a nitrogen atmosphere and then concentrated in vacuo to a yellow syrup. The remaining hydrochloric acid was removed by co-distillation with water (2 x 10 mL). The crude material was purified by transfer column chromatography (2.2 x 13.0 cm PrepPak C-18) with water as the eluting solvent. The appropriate fractions were combined and lyophilized to give a white powder; 1.43 g, 85.4%; infrared (KBr) ^: 2565 (SH),

, c —1 maX

1626 (pyridinium), 1585 (pyridinium) cm; uv (H 2 O) ν max: 278 (7355); Anal. determined values for C 9 H 12 N 2 O 3 S 2 F 3: C 35.64, H 3.99, N 4.62, S 21.14; found values; C 35.49, H 4.05, N 4.56, S 20.99.

30 1 149 7 8 0 9 4 B. (5R, 6S) 3 - [(1, e-Dimethylpyridinium 4-yl] methylthio] - [1- (1R-hydroxyethyl) -4R-methyl-7-oxo-1-azabicyclo - / ~ 3 2. Q7hept-2-ene-2-carboxylate 5 o OH CH3 1- (PhO) 2PCl OH CHj jMj "'" i> M / c »<Sr \ _ JJ /„ 3- Thiol * Buffer- * 3 II 3

0 ^ - 4-HEt (iPr) 2 solution 7.0 0 <** - | COJ

coopnb Eetheri coö ^ ΤΗΡ

1C

To a cold solution (5 ° C) of (5R, 6S) paranitrobenzyl-3,7-dioxo-6- (1R-hydroxyethyl) -4RS-methyl-1-azabicyclo [1.2.0] heptane-2 R-carboxylate (1.11 g, 3.06 15 mmol, R / S: 86/14) in dry acetonitrile (90 mL) maintained under a nitrogen atmosphere was added simultaneously with diphenyl chlorophosphate (0.68 mL, 3.3 mmol) and diisopropylethylamine (0.57 mL, 3.3 mmol) over 10 minutes. The cold (5 ° C) mixture was stirred for 1 h, cooled to 20-30 ° C and treated simultaneously with a solution of (1,6-dimethylpyridinium-2-yl) methylthiol, trifluoromethanesulfonate salt (1.03 g, 3, 4 mmol) in dry acetonitrile (2 mL) and diisopropylethylamine (0.59 mL, 3.4 mmol) over 15 minutes. The resulting mixture was stirred for 0.5 h at -30 ° C, warmed to 0 ° C and stirred for 1.0 h before being diluted with cold water (35 mL). The resulting emulsion was poured onto the top of a phase transfer column (PrepPak C-18, 2.5 x 18 cm), and the column was eluted with a mixture of 25-50% acetonitrile in water. The appropriate fractions were lyophilized to give a sticky yellow solid, 1.69 g, which was dissolved in wet tetrahydrofuran (40 mL). To the resulting solution were added ether (70 ml), potassium dihydrogen phosphate-sodium hydroxide buffer solution (pH 7.0, 0.2M, 50 ml) and 10% palladium on carbon 35 (1.69 g), and the resulting mixture was hydrogenated at 290 kPa: at 23 ° C for two hours and then filtered through a diatomaceous earth filter.

150 7 8 0 9 4

The phases were separated and the aqueous phase was washed with ether (2 x 20 mL) and concentrated under high vacuum at <23 ° C to 15 mL placed on top of the phase transfer column (prepPak C-18). Elution with a mixture of 4% acetonitrile in water gave, after lyophilization of the appropriate fractions, 0.23 g of the title product mixed with potassium sodium diphenyl phosphate (24% in moles). Re-purification on a reverse phase column (2.5 x 14 cm, prepPak C-18) of water (400 ml) and a mixture of 10% acetonitrile in water (200 ml) as eluent gave lyophilization of the appropriate fractions. followed by a yellow powder, 0.17 g, 15.3%; infrared (KBr) Ό _ s 1750 (β-lactam C = 0), 1625 (pyridinium), ΠΙαΧ _ - m 1600 (carboxylate C = 0) cm; Hnmr (D 2 O) δ: 1.12 (d, J = 7.2 Hz, CH 3 in C-4), 1.24 (d, J = 6.4 Hz, CH 3 CHOH), 2.80 ( s, CH 3 in pyridinium C-6), 4.18 (CH 3 in pyridinium N), 4.41 (AB quartet center, CH 2 S), 7.5-8.4 (Hs in pyridinium); uv (buffer solution 0.05M, pH 278 (S 11504); [1,256.4 ° (c 0.22, Η2 ° ^; Ί ^ 1/2 = hours measured at 37 ° C in buffer solution (pH 7.4)) to a concentration of 10 ~ ^ M.

20

Example 26 (5R, 6S) -3- [1- (1,6-dimethylpyridin-2-yl) methylthio] -6- (1R-hydroxyethyl) -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-25 Preparation of 2-carboxylate CH3 30 'o

> j, ^ r i®J

cbo Θ 35 151 78094 A. (5R, 6S) -paranitrobenzyl 11-3 - [(1,6-dimethylpyridinium-2-yl) methylthio] -6- (1R-hydroxyethyl) -7-oxo-1-azabicyclo /3.2 Q7hept-2-ene-2-carboxylate, trifluoromethanesulfonate salt and diphenylphosphate salt.

5

MeOTi 1- NaOH vs.

"Eetterl>" Λ '. P

3 I

10 CT3 1- (PhO) 2POCl | H c »3 1t. 0 1 4- NEt (iPr) j] ^ (Ph0,2POp 1 5 COOPNB * C00PN8 2 2

To a cold (5 ° C) solution of (5R, 6S) -paranitro-20 ° benzyl 6- (1R-hydroxyethyl) -3,7-dioxo-1-azabicyclo [3.2.0] heptane-2R-carboxylate ( 2.14 g, 6.14 mmol) in dry acetonitrile (18 mL) and maintained under a nitrogen atmosphere, diphenyl chlorophosphate (1.37 mL, 6.6 mmol), diisopropylethylamine (1.15 mL, 6.6 mmol) was added. mmol) in such amounts that the temperature was maintained at 5 ° C (7-10 min) and 4-dimethylaminopyridine (6 mg, 0.05 mmol). The mixture was stirred for 1.5 hours at 5 ° C, and used as such; this mixture will be referred to as "solution A" later in the process. A solution of (6-methylpyridin-2-yl) methylthioacetate-30 (1.23 g, 6.8 mmol) in dry ether (10 mL) maintained under a nitrogen atmosphere was treated with methyl triflate (0.85 mL, 7.5 mmol) and stirred for 1.5 hours at 23 ° C. The ether was decanted and the white powder was washed twice with ether (2 x 10 mL) and dissolved in water (10 mL). The resulting aqueous solution was cooled to 0 ° C under an oxygen-free atmosphere and treated with sodium hydroxide (4N, 3.4 mL, 13.6 mmol).

152 78094

The mixture was stirred at 2 ° C for 1 hour, and then the pH was adjusted to 7.6 by the addition of potassium dihydrogen phosphate; this mixture will be referred to as "solution B" later in the process. Cold (5 ° C) "solution A" was treated with "solution B" for 0.5 to 5 hours, during which time the pH was maintained between 7.25 and 7.35 by dropwise addition of 4N sodium hydroxide solution. The mixture was stirred for 0.5 h and poured onto the top of a reversed phase column (4.0 x 18 cm, prePak C-18); the column was eluted with a mixture of 25-50% acetonitrile in water. Lyophil-10 ringing the appropriate fractions gave the title product as a yellow powder, 2.82 g (51% (PhO) 2 PO 2 -, 49%, CF 2 SO 4), 80%; infrared (KBr) ^: 3700-3000 (OH), 1772 ((β-lactam C = 0), max 1700 (ester C = 0), 1625 (pyridinium), 1590 (pyridinium) cm-1; 1Hnrar ( DMSO, d-6) δ 1.15 (d, J = 6.2 Hz, CH-jCHOH), 2.84 (s, CH 2 pyridinium in C-6), 4.16 (s, CH 2 (pyridinium in N), 4.79 (p. SCH 2), 6.6-7, S / 'iPhO) 2 PO 2 -7, 7.5-8.7 (pyridinium hydrogens and PNB ester hydrogens).

B. (5R, 6S) 3 - [(1,6-Dimethylpyridinium-2-yl) methyl-thio] -6- (1R-hydroxyethyl) -7-oxo-1-azabicyclo / 3.2.Q7hept-2-ene -2-carboxylate, COOPNB solution

COO

To a solution of (5R, 6S) paranitrobenzyl-3-Z- (1,6-dimethylpyridinium-2-yl) methylthio] -6- (1R-hydroxyethyl) -7-oxo-1-azabicyclo [2.2. Q7hept-2-ene-2-carboxylate, trifluoromethanesulfonate and diphenylphosphate salt (49:51) (0.87 g, 1.27 mmol) in wet tetrahydrofuran (50 mL) was added ether (50 mL), potassium dihydrogen phosphate-potassium hydroxide spray solution (0.1M, 40ml, pH 7.0) and 10% palladium on carbon (0.87 g). The mixture was hydrogenated at 248 kPa at 23 ° C for two hours and filtered through a diatomaceous earth filter.

The phases were separated and the aqueous phase was washed with ether (2 x 15 ml, concentrated under high vacuum to 30 ml and poured onto the top of 5 reverse phase columns (PrepPak C-18, 2.2 x 13 cm). The column was eluted with water. The appropriate fractions were combined and lyophilized, to give a yellow powder, 0.719 g, 40%, infrared Λ max! 1755 (β-lactam C = 0), 1628 (pyridinium), 1590 (carboxylate C = 0) cm -1 1Hnmr (D2O) <$: 1, 25 (d, J = 6.4 Hz, CH-jCHOH), 2.82 (s, CH 3 in pyridinium C-6), 3.12 ('dd', J = 9.2 Hz, J = 2.9 Hz, H-4), 3.39 (dd, J = 6.0 Hz, J = 2.8 Hz, H-6), 3.7-4.4 (CH-jCHOH, H-5 , CH3 in pyridinium N), 4.48 (s, C ^ S), 7.6-8.4 (pyridinium hydrogens), uv (HO) A: 279 (C 9628) with protrusion at 296; λ λ ft ΧιαΧ 15 £ <XJd 55.0 C (c 0.63, H 2 O); ^ j2 ~ "* 2.5 hours measured at 37 ° C in a buffer solution of pH 7.4 to a concentration of 10-4M.

Example 27 3- [2- (N-Methylpyridinium) methanethio] -6 ° C - [1- (R) -hydroxyethyl] -4H-methyl-7-oxo-1-azabicyclo(3.2.0)hept-2 Preparation of -ene-2-carboxylate.

OH cg ol2 Θ ch3 30 n2 8 Ä 1 3 4 / Λ / 35 154 78094 A. p-Nitrobenzyl 2-diazo-3-oxo-n-valerate (4).

A solution of 50 g (0.35M) of ethyl 3-oxo-n-valerate and 54 g (0.35M) of p-nitrobenzyl alcohol in 400 ml of toluene was heated at 130-140 ° C without reflux. 5 teachers for 18 hours. The solvents were evaporated to give a yellow crystalline solid which was recrystallized from Et 2 O-pentane to give 75 g (86% yield) of p-nitrobenzyl 3-oxo-n-valerate (3).

mp: 33-34 °. IR (KBr) 1740 and 1705 cm-1. NMR (CDCl 3) δ 1.20 (3H, t, H = 7.0Hz), 2.65 (2H, q, J = 7.0Hz), 3.60 (2H, s), 5.28 (2H , s), 7.45 (2H, d, J = 9.5Hz), and 8.18 (2H, d, J = 9.5Hz). To a solution of 55.5 g (0.22M) in 5003,500 mL of CH 2 CN was added

O

first at 0 ° C 45 g (0.44M) of TEA and then 50 g (0.22M) of p-carboxybenzenesulfonyl azide. The ice bath was removed and the mixture was allowed to stir for 90 minutes. The precipitate was filtered, washed with CH 2 Cl 2 and the filtrate was concentrated to about 100 mL and diluted with 800 mL of EtOAc. The organic solution was washed with aqueous NaHCO 3, salted and dried (MgSO 4). The dried solvents were evaporated to give 55 g (90% yield) of compound as yellowish crystals, m.p. 96-97 °. IR (KBr) V 2120 and 1710 cm-1. NMR (CDCl 3) δ 1.20 (3H, t, J = 7.0Hz), 2.85 (2H, q, J = 7.0Hz), 5.40 (2H, s), 7.50 (2H, d, J = 8.0Hz), and 8.15 (2H, d, J = 8.0Hz).

25 OSi

--- »\ ^ \ ^ C02PNB

30 N2 N2 4 5 35 155 78094 B. 1-p-Nitrobenzyloxycarbonyl-1-diazo-2-t-butyldimethylsilyloxy-2-butene (5).

To a cooled (0 ° C) solution of 54 g (0.2M) of Compound A in 400 mL of Cl 2 Cl 2 was added 41.4 g (0.4M) of TEA followed by 56 g (0 M) of TEA. , 21M) tert-butyldimethylsilyl chloride in 30 mL of CH 2 Cl 2 for 40 minutes. The solution was stirred for 120 minutes, then washed with ice water. The CH 2 Cl 2 was dried (MgSO 4), filtered and evaporated in vacuo to give 68 g (89% yield) of 5 as a yellow solid. mp: 54-55 °. IR (KBr) γ 2080 and 1695 cm. NMR of compound i showed that compound i> was obtained as an E / Z mixture in the olefin position in a ratio of 9: 1. NMR (major isomer of CDCl 3) δ 0.15 (6H, s), 0.90 (9H, s), 1.58 (3H, d, J = 7.0 Hz), 5.15 (2H, s), δ , 30- (2H, d, J = 9.0Hz) and 8.0 (2H, d, J = 9.0Hz).

15 I Ö5i + ° | S | i + OSi + ^ CH, N.

[? (2PSB -> 6 5 25 C. 4 - [(3-1-methyl-3-diazo-3-p-nitrobenzyloxycarbonyl-2-oxopropyl) -3- [- (R) -t- butyldimethylsilyloxyethyl 147-azetidin-2-one.

To a suspended solution of 12.5 g (0.1M) of anhydrous ZnCl 2 in 700 mL of CH 2 Cl 2 was added 60.4 g (0.21M) of E and stirred for 15 minutes at 23 ° C and then cooled to 0 ° C. A solution of 106 g (0.27M) of J5 in 200 ml of CH 2 Cl 2 was added dropwise to the above reaction solution over 90 minutes, then stirred for 120 minutes without a cooling jacket. The reaction mixture was washed with aqueous NaHCO 3 (4 x 150 mL), water, brine and dried (MgSO 4). Evaporation of the dried solvents gave a dark 156 7 8 0 9 4 oil which was purified on a SiO 2 column; column elution was performed with EtOAc-CH 2 Cl 2 (1: 9) to give 51.5 g (54%) of J. as a white crystalline solid. mp: 112-114 ° C. IR (KBr) Y 2130, 1760 and 1720 cm -1. The NMR of compound 760 at 5 MHz showed that compound 7 was obtained as a mixture in the 1-methyl position in a ratio of 2: 1. NMR (CDCl 3) δ 0.3-0, Δ (6H, 2s), 0.8Z (9H, 2s), 1.05-1.15 (6H, m), 2.68 (0.66H, q, J = 6.6 and 2.0Hz), 2.88 (0.34, q, J = 6.6 and 2.0Hz) 3.57 (1H, m), 3.84 (1H, m), 4.09 (1H, m), 517 (2H , two), 5.84 (0.66H, s), 5.95 (0.34H, s), 7.52 (2H, d, J = 8.5 at 10 Hz) and 8.23 (2H, d). , J = 8.5Hz).

»^ Jo.

✓ “« · Πη Ϊ C02PNB

20 D. 4/3- (1-Methyl-3-diazo-3-p-nitrobenzyloxycarbonyl-2-oxopropyl) -3 c * - [1- (R) -hydroxyethyl] -azetidin-2-one ( 8).

To a solution of 30 g (59.5 mmol) of the compound in 7,400 mL of MeOH was added 150 mL of 1N HCl at 23 ° C and stirred for 18 h. The reaction solution was concentrated to about 200 mL and extracted with EtOAc (3 x 200 mL). The combined EtOAc was washed with water, aqueous NaHCO 3 and salted. Evaporation of the dried (MgSO 4) solvents gave 22.3 g (96%) of compound 3 as a white crystalline solid, mp 147-148 ° C. IR (KBr) - 3400, 2135 and 1750 cm -1. 360 MHz nmr showed that compound j3 was obtained as a mixture in the 4-methyl position in a ratio of 2: 1. NMR (DMSO-d) 1.07-1.10 (6H, m), 2.75 (0.66H, q, J = 6.6 and 2.0Hz), 2.85 (0.34H, J = 6.6 and 2.0Hz), 3.55-3.90 (3H, m), 35.25 (2H, s), 7.70 (2H, d, J = 9.0 Hz), 8.05 (0.66H, s), 8.10 (0.34H, s) and 8.27 (2H, d, J = 9.0Hz).

li 157 78094

C02PNB

As As 10 E. p-Nitrobenzyl 6-β-β- (R) -hydroxyethyl] -4-methyl-3,7-dioxo-1-azabicyclo (3.2.07 heptane-2-carboxylate (9)).

A solution of 14.0 g (35.86 mmol) of 8 and 70 mg of rhodium (II) octanoate in ethyl acetate was heated at reflux for 20 minutes in the presence of nitrogen. The mixture was evaporated in vacuo to give compound 9. IRlCHCl 3 * 3400 and 1750 cm -1. The 360 MHz nmr of compound 5) showed that compound 9 was obtained as a mixture in the 4-methyl position in a ratio of 2: 1. Overhauser nuclear effects (NOE) were used as an aid in determining the 4-methyl configuration. When the H 1 of the main isomer was irradiated, an estimated 7% more signals of 4-methyl protons were detected, indicating a cis linkage between H 2 and 4T-methyl. On the other hand, when the H 2 of the side isomer was irradiated, no signal amplification for 4-methyl was observed, indicating a trans linkage between the H 1 of the side isomer and 4-methyl.

NMR (CDCl 3) for the major isomer δ 1.24 (3H, d, J = 7.35Hz), 1.40 (3H, d, J = 6.3Hz), 2.40 (1H, m), 3.24 (1H , q, J = 6.6 and 7.2Hz), 3.67 (1H, q, J = 8.0 and 2.2Hz), 4.18 (1H, m), 4.82 (1H, s) , 5.24 (1H, d, J = 6.3Hz), δ 6.18 (1H, d, J = 6.3Hz), 7.60 (1H, d, J = 8.5Hz) and 8.22 (1H, d, J = 8.5H).

NMR (CDCl 3) for side isomer S 1.0 (3H, d, H = 7.35Hz), 1.40 (3H, d, J = 6.3Hz), 2.83 (1H, m), 3.25 ( 1H, q, J = 6.6 and 1.50Hz), 4.14 (1H, q, J = 7.36 and 1.50Hz), 4.67 (1H, s), 5.24 (1H, d , J = 6.3Hz), 6.18 (1H, d, J = 6.3Hz) and 7.60 (1H, d, J = 8.5Hz) and 8.22 (1H, d, J = 8, 5Hz).

35 158 78094 5 0ίΧτ ^} J-Iy ^ -P (o0 ^

cb2PNB C02PNB

^ 10, 10 F. p-Nitrobenzyl-3-diphenoxyphosphinyl-6- [1- (R) -hydroxyethyl] 117-4 / 3-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene- 2-carboxylate (10).

To a cooled (0 ° C) solution of 20.0 g (55.2 mmol) of the keto intermediate in 9,150 mL of CH 2 CN was added 7.18 g of 15 6 (55 mmol) of diisopropylethylamine and its then 14.85 g (55 mmol) of diphenyl chlorophosphate in 20 ml of CH 2 Cl 2 for 5 minutes. The resulting solution was stirred for 60 minutes at 0 ° C, then diluted with 600 mL of EtOAc, washed with ice-cold 10% H 2 PO 4 and brine. Haih-20 addition of dried (MgSO 4) solvents gave an oil which was purified on a SiO 2 column; the column was eluted with a solution of 10% EtOAc in CH 2 Cl 2 to give 3.7 g (11.5%) of the phosphonate as a white form. IR (CHCl 3) Y 3400, 2790 and 1720 cm -1.

NMR (CDCl 3) δ 12.0 (3H, d, J = 7.2 Hz), 1.38 (3H, d, H = 7.3 Hz), 3.35 (111, J = 6.7 and 2.0Hz), 3.50 (111, m), 4.2-4.25 (2H, m), 5.20 (1H, d, J = 10.5Hz), 5.37 (1H, d) , J = 10.5Hz), 7.1-7.4 (1OH, m), 7.56 (1II, d, J = 9.0Hz) and 8.10 (1H, d, J = 9.0Hz) ). NOE was used to determine the 4-methyl configuration of compound H). When H 2 was irradiated for 30 s, no signal amplification was observed for 4-methyl, indicating trans-configuration of H 2 and 4-methyl.

159 78094 0Η / Ν * Λ ^ 'r-Λ s * S ^ Ö.

J3> O-P (O0) 2 ->

5 0 COjPNB

COjPNB

10 11 10 G. p-Nitrobenzyl-3- (pyridin-2-yl-methanethio) 6- [1- (R) -hydroxyethyl] -4-methyl-7-oxo-1-azabicyclo- (3.2.0) hept -2-ene-2-carboxylate (11).

To a cooled (-15 ° C) solution of 1.2 g (2 mmol) of phospho-1H-phosphate in 10 mL of CH 2 Cl 2 was added 390 mg (3 mmol) of diisopropylethylamine followed by 370 mg (3 mmol) of diisopropylethylamine. mmol) of 2-mercaptomethylpyridine in the presence of nitrogen. The reaction mixture was allowed to stir for 60 minutes at -15 ° C, then for another 60 minutes. 0 ° C. The reaction mixture was diluted with EtOAc, washed with ice water, salted and dried (MgSO 4). The solvents were evaporated in vacuo to give a yellow oil which was purified on a SiO 2 column; the column was eluted with a solution of 20% EtOAc in CH 2 Cl 2 to give 375 mg (40% yield) of _1_1_ as a white amorphous foam. IR (KBr) v * 3400, 1775 and 1710 cm-1. NMR (CDCl 3) δ 2.14 (3H, d, J = 6.7Hz), 2.19 (3H, --d, J = 6.7Hz), 3.14 (1H, q, J = 6.2 and 2.0Hz), 3.40 (1H, m), 4.0 (1H, d, -J = 7.6Hz), 4.12 (1H, d, J * 7.6Hz), 4.18 (1H, q, J = 6.7 and 2.0Hz), 4.25- (1H, m), 5.25 (1H, d, J = 11.3Hz), 5.40 (1H, d , J = 11.3 Hz), 7.15-8.2- (4H, m).

30 160 78094

OH

5 Λ ^^ Τ3 f CH3

o - * 'oV'O

do.PNB S [/ CO 2 CH 3® 10 H. 3- / 2- (N-Methylpyridinium) methanethio7-6 c- [1- (R) -hydroxyethyl] -4 / 3-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (12).

15

To a solution of 1.0 g (2 mmol) of the compound in 10 mL of CH 2 Cl 2 was added 450 mg (3.3 mmol) of methyl trifluoromethanesulfonate and stirred at 23 ° C for 90 minutes. Evaporation of CH 2 Cl 2 in vacuo gave carvenized pyridine-20 as a foam which was immediately hydrated without further purification. The crude pyridinium salt was dissolved in TMF-ether-PH 7 buffer solution (1: 1: 1, 100 ml each) and then 600 mg of 10% palladium on carbon was added. The mixture was hydrogenated at 240 kPa with pair stirrers for 45 min. The mixture was filtered through a pad of diatomaceous earth and the catalyst was washed with water (2 x 10 mL). The combined filtrates and washings were extracted with ether (2 x 100 mL) and lyophilized to give a yellow powder which was purified on a C 18 g BONDPAK reverse phase column (10 g), eluting with a solution of 5% -CH 2 CN in water, 55 at a pressure of kPa. Each 15 ml fraction was analyzed by high performance solution chromatography, and fractions with ultraviolet absorption at a maximum of 300 nm were collected and lyophilized to give 58 mg (11% yield) of the title product as a pale yellow powder. IR (KBr) λ max 3400, 1750 and 1590 cm -1 35% max (H 2 O) 292 nm (£ 7081). NMR (D 2 O) δ 1.1 (3H, d, J = 6.5Hz), 1.23 (3H, d, J = 6.5Hz), 3.18 (1H, m), 3.45 (1 H, q, J = 6.0 and 2.1Hz), 4.0-4.4 (4H, m), 4.65 (3H, s), 7.79 (2H, m), 8.30 ( 1H, m) and 8.60 (1H, m).

161 78094

Example 28 3- [2- (1,4-Dimethylpyridinium) methanoyl] -6 (N- (1-1 (R) -1-hydroxyethyl) -4H-methyl-7-oxo-1-azabicyclo (3.2.0) hepto- Preparation of 2-ene-2-carboxylate.

5 il OB C = 3 1) BS '/ V | 1 OB CH, f 10] χ ^ - ° -? <Οι9,2 a] x> s ^ 4 0 COjPHB -> ° C0j® 3) P «VH2 10 13 15 3- [2- (1,4-Dimethylpyridinium) methanethio] -6 <X. - [1- (R) -20 hydroxyethyl] -4 / 3-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (13).

This compound was obtained as a yellow powder in 17% yield from compound 10 in the same manner as in Example 27. IR Y 3400, 1755 and 1600 cm-1. UV λ max (H 2 O) 300 nm 25 (£ 7600). NMR (D 2 O) δ 1.20 (3H, d, J = 6.7Hz), 1.28 (3H, d, J = 6.7Hz), 2.60 (3H, s), 3.4-3, Δ (2H, m), 4.2-4.4 (4H, m), 4.52 (3H, s), 7.82 (1H, t, J = 6.5 and 4.2Hz), δ, 32 (1H, d, J = 6.5Hz) and 8.60 (1H, 6, J = 4.2Hz).

Example 29 3- [4- (1-Methylpyridinium) methanethio] -6 - (- [1- (R) -hydroxyethyl] -4H-methyl-7-oxo-1-azabicyclo (3.2.0) hept-2- preparation of ene-2-carboxylate.

35 78094 162 1) BS ΛΌ »α OH CH- 2) CF3SO3CH, 1 jr -. - "T 31 Pd / H2 co2®

COjPHB

10 10 14 14 3- [4- (1-methylpyridinium) methanethio] -6 cX- [1- (R) -hydroxyethyl] -4,3-methyl-7-oxo-1-azabicyclo (3.2.0) hept - 15 2-ene-2-carboxylate (14).

This compound was obtained as a yellow powder in 15% yield from compound H) in the same manner as in Example 27. IR (KBr) Y3410, 1750 and 1650 cm-1. UV λmax (H2 O) 293 nm (£ 7295). NMR (D_0) Δ 1.15 (3H, d, J = 6.5 Hz), 1.20 (3H, d, J = 6.5- ΟΛ ^

Hz), 3.20 (1H, m), 3.45 (1H, q, J = 6.0 and 2.0Hz), 4.11 (1H, q, J = 8.0 and 2.0Hz) , 4.20 (1H, m) and 4.35 (3H, s), 7.95 (2H, d, J = 5.2Hz) and 8.72- (2H, d, J = 5.2Hz).

Example 30 3-Z3- (1-Methylpyridinium) methanethio7-6- [1- (R) -hydroxyethyl] 47-4 / 3-methyl-7-oxo-1-azabicyclo (3.2.0) hept Preparation of -2-ene-2-carboxylate.

30 hsA ^ 3 CH

OH CH3 2) CF3SO3CH3

A Jl n V

-3) 0

° CO, PNB

35 2 15 rw 10 163 7 8 0 9 4 3- [3- (1-Methylpyridinium) methanethio] -6 ° C / 1- (R) -hydroxyethyl] -4,3-methyl-7-oxo-1-azabicyclo (3.2. 0) hept-2-ene-2-carboxylate (15).

This compound was obtained as a yellow powder in 27% yield-zero from compound 1j) in the same manner as in Example 27. IR (KBr) V 3420, 1750 and 1610 cm-1. UV λmax (H 2 O) 295 nm (ε 8750) NMR (D 2 <0) δ ≤ 10 (3H, d, J = 6.9Hz), 1.25 (3H, d, 9Hz), 1.27 ( 1H, m), 1.43 (1H, q, J = 6.2 and 1.8Hz), 4.1-4.35 (4H, m), 4.39- (3H, s), 8.0 (1H, t, J = 8.5 and 6.2Hz), 8.45 (1H, d, J = 8.5Hz), 8.70-10 (1H, d, J = 6.2Hz) and 8, 82 (1H, s).

Anal. Calculated values 2-20: C: 5190; H, 6.36; N, 7.12 Found: C, 51.92; H, 5.71; N, 6.88.

Example 31 3- [3- (1/2-Dimethylpyridinium) methanethio] -6 c- [1- (R) -hydroxyethyl] -4-4-methyl-7-oxo-1-azabicyclo (3.2.0) hept - Preparation of 2-ene-2-carbosylate.

20 1) HS ^ v ^ «ffl3 2) CP, SO3CH3 25 JL JL O V fr-x-V VT» / iä -3, Pa ^ °

0 COjPKB

16 30 10 * "* - 35 164 7 8 0 9 4 3- [3- (1,2-Dimethylpyridinium) methanethiol7-6 c * Z-1- (R) -hydroxyethyl] -4C-methyl-7- oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (16).

This compound was obtained as a yellow powder with 14% yield-zero from compound JU) in the same manner as in Example 27. IR (KBr) Y3410, 1750 and 1600 cm -1 UV λ max (H 2 O) 296 nm (£ 8500). NMR (D 2 O) δ 1.12 (3H, d, J = 6.5 Hz), 1.30 (3H, d, J = 6.5 Hz), 2.95 (3H, s), 3.40 ( 1H, m), 3.50 (1H, q, J = 6.2 and 1.8 Hz), 4.24-4.4 (4H, m), 4.35 (3H, s), 7.82 ( 1H, t, J = 8.5 and 6.3Hz), 8.40 (1H, d, J = 8.5Hz) and 8.72 (1H, d, J = 6.3Hz).

Example 32 / 3- (2,4-Dimethyl-1,2,4-triazolium) methanethiol-6- [1- (R) -hydroxyethyl] -4- (2'-methyl-7-oxo-1 Preparation of -azabicyclo- (3.2.0) hept-2-ene-2-carboxylate.

f3

20 ^ y-Λ, <? L

iH ^ T ° -p (0 | i> 2 21 = 3 ^ 3 = ¾ nQ ^^ jj “a» -> 0 I® i, 3) pä / a2 2 10 25 ~ 17 ^ / ~ 3- (2 , 4-Dimethyl-1,2,4-triazolium) methanethio-7- (β- (1-) - (R) -hydroxyethyl-7- [4- (4-methyl) -7-oxo-1-azabicyclo [3.2.1] hept- 2-ene-2-carboxylate (17).

This compound was obtained as a yellow powder in 9% yield from _1 () in the same manner as in Example 27. IR (KBr) / 3420, 1756 and 1605 cm-1. UV λ max (H 2 O) 291 nm (£ 7850). NMR (D_O) δ 1.15 (3H, d, J = 6.3 Hz), 1.22 (3H, d, J = 6.3 35 2 Hz), 3.35 (1H, m), 3.48 (1H, q, J = 6.0 and 1.8Hz), 3.90 (3H, s), 4.05- (3H, s), 4.2-4.4 (4H, m) and 8, 80 (1H, s). 1,165,78094

Example 33 3- [2- (1,3-Dimethylimidazolium methanethio] 6- [1- (R) -hydroxyethyl] -r3'-methyl-7-oxo-1-azabicyclo (3.2.0) hept-5 2-ene-2 carboxylate preparation T3 10 0H w ® jl ® 21 w ·

Cl I X o> A rL

CO 2 PNB -> 00 ^ “3 3) Pd / Hj 15 10 18 3- [2- (1,3-Dimethylimidazolium methanethyl) -6 cis - (R) -hydroxyethyl] -4,7-methyl-7- oxo-1-azabicyclo (3.2.0) hept-2-ene-2-carboxylate (18).

This compound was obtained as a yellow powder in 32% yield from 1J) in the same manner as in Example 27. IR (KBr) γ, 3400, 1758 and 1600 cm -1. UV λ max (H2O) 294 nm (E 7194). NMR <D 2 O) δ 1.10 (3H, d, J = 6.3 Hz), 1.25 (3H, d, J = 6.3 Hz), 3.30 (1H, m), 3.42 ( 1H, 1, J = 6.0 and 2.2 Hz), 3.85 (6H, s), 4.2-4.6 (4H, m) and 7.40 (2H, s) 166 78094

Example 34 (5R, 6S) -6- (1R-hydroxyethyl) -4R-methyl-3 - [(3-methyl-1,2,3-thiadiazolium-4-yl) methylthio] -7-oxo-1 -aza-bicyclo [3.2.0] hept-2-ene-2-carboxylate (BMY-26659) _ 5 OH ς

fvN

i-N | + 01-CH, to2 3 10 (5R, 6S) -paranitrobenzyl-6- (1R-hydroxyethyl) -4R-methyl-3 - [(1,2,3-thiazol-4-yl) methylthio] -7-oxo -1-azabicyclo | 3.2.0 | hept-2-ene-2-carboxylate_ HO ** 3 1 fj 15 7-o - [- λ.

oJT> -o?, o0,2 -► Λ "'/' 5 <3

I CO-PNB X V

co2pnb 2 (5R, 6S) -paranitrobenzyl-6- (1R-hydroxyethyl) -3-20 (diphenylphosphono) -4R-methyl-7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2- a cold solution of the carboxylate (3 mmol) (5 ° C) in acetonitrile (12 mL) was treated with 4 mercaptomethyl-1,2,3-thiadiazole (528 mg, 4 mmol) and diisopropylethylamine (426 mg, 3.3 mmol, 0.58 ml). After stirring for 25 L, the solution was diluted with ethyl acetate (35 mL) and washed twice with cold water. The organic solution was dried (MgSO 4) and evaporated and the crude product was passed through an ultra-pure SiO 2 layer (40 g). Elution was performed with diethyl ether (10 x 20 mL) followed by ethyl acetate 30 (10 x 20 mL). Evaporation of the relevant fractions gave 1.18 g (82.5%) of a pale yellow foam. IR (Cl 2 Cl 2) ν: 3600 (-OH), 1775 (β-lactam), 1712 cm -1 (ester].

ITlclX

1 H-NMR (CDCl 3) δ: 8.45 (1H, s, Ar-H), 8.21 (2H, d, Hm aro-J max aromatic), 7.63 (2H, d, Ho aromatic), 5.34 (2H, ABq, 35 H-benzyl), 4.54 (2H, ABq, -SCH 2 -), 4.4-4.0 (2H, m, H-1 ', H-5), 3 , 71 (1H, m, H1), 3.28 (1H, dd, J5_g = 2.5, Jlt_g = 6.7 Hz, H-6), 1.75 (1H, broad s, OH), 1, 35 (3H, d, J1_2 · = 6.1 Hz, H-2 '), 1.30 (3H, d, J1_CH3 = Hz (1-CH.j)).

5,167,78094 (5R, 6S) -6- (1R-hydroxyethyl) -4R-methyl-3 - [(3-methyl-1,2,3-thiadiazolium-4-yl) methylthio] -7-oxo-1 aza-bisykloCs. 2. Oj hept-2-ene-2-carboxylate_

Hp CH3 ho ch3 ho ch3 ^ λ — 0 / CH3 / H3 co2pnb CO2pnb S toP s 10 (5R, 6S) -paranitrobenzyl-6- (1R-hydroxyethyl) -4R-methyl-3-Q (1,2,3- thiadiazol-4-ylmethylthio} -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (2.25 g, 4.72 mmol) in CH 2 Cl 2 (14 mL) and diethyl ether (9 mL). Methyl trifluoromethanesulfonate (775 mg, 4.72 mmol) was added dropwise at room temperature, the solution rapidly clouded and an oil precipitated, the mixture was stirred for 2 hours, then the mother liquor was decanted off and the oil was triturated with diethyl ether to give a yellow powder (2, 08 g, 68.8%) The quaternized carbapenem (1.0 g, 1.56 mmol) was dissolved in THF (25 mL), diethyl ether (25 mL) and phosphate buffer (0.2 M, pH 7, 16 mL). and hydrogenated for 75 minutes in the presence of 10% Pd.C (1.0 g) at 310 kPa 25. The reaction mixture was then cooled on ice, filtered and the filtrate was extracted twice with diethyl ether. with donkey SiO 2 eluting with water and then with a 5% mixture of CH 2 Cl 2 and water. The above hydrogenation was repeated under the same conditions and to the same extent. The two batches were combined and the product was rechromatographed on reverse phase SiO 2. H 2 O (175 mL) and 5% CH 2 Cl 2 in H 2 O were used for elution. The appropriate fractions were combined and lyophilized to give 164 mg (14.8%) of carbapenem. IR (nujol)

35 V: 1750 (β-lactam), 1590 cm -1 (carboxylate). 1 HMR

(D 2 O) δ: 4.64 (3H, s, N -CH 2 -), 4.50 (2H, ABq, -S-CH 2 -), 4.29-4.11 (2H, m, H 1 ', H -5), 3.48 (1H, dd, J 5 -6 = 2.7, = 168 7 8 0 9 4 5.8 Hz, H-6), 3.35 (1H, m, H1), 1.29 ( 3H, d, = 6> 3

Hz, H-2 '), 1.20 (3H, d, J 1 -CH 3 = 7.1 Hz, 1-CH 3); UV H 2 O: 294 (7764).

Example 35 5 (5R, 6S) -6- (1R-hydroxyethyl) -4R-methyl-3 - [(1,3-dimethyl-1,2,3-triazolium-4-yl) methylthio] -7-oxo- 1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (BMY-26873) CH3 3 z CH3 15 (5R, 6S) -paranitrobenzyl-6- (1R-hydroxyethyl) -4R-methyl-3 - [(1-methyl-1,2,3-triazol-4-yl) methylthio] -7-oxo-1-azabicyclo [3.2.0] hept-2-ene-2-carboxylate (1.1 g, 2.32 mmol) ) to a solution in CH 2 Cl 2 (6 mL) and diethyl ether (5 ° F) was added methyl trifluoromethanesulfonate (419 mg, 2.55 mmol, 0.29 mL) dropwise at room temperature. The oil separated immediately. After stirring for one hour, the mother liquor was decanted and the oil was triturated with diethyl ether to give a slightly yellowish powder; yield 1.31 g (88.6%).

The quaternized compound (1.02 g, 1.6 mmol) was dissolved in a mixture of THF (25 mL), diethyl ether (25 mL) and phosphate buffer (1.2 M, pH 7.0, 16 mL) and hydrogenated for 60 minutes. in the presence of% Pd-C (1.0 g) at 276 kPa. The reaction mixture was then cooled on ice, filtered and the filtrate was extracted with diethyl ether (2 x 20 mL). The aqueous solution was chromatographed on reverse phase SiO 2 (20 g); cold (5 ° C) water (200 mL) and 5% CH 2 Cl 2 in H 2 O (100 mL) were used for elution. The relevant fractions were pooled and lyophilized. Yield 320 mg (56.9%). IR (nujol) v: 1 1 rnäx r 35 1751 (β-lactam), 1600 cm (carboxylate). MR (DMSOd) 8.90 (1H, s, Ar-H), 4.55-3.75 [4H, complex group, H-1 ', H-5, -S-CH2), 4.27 ( 6H, s, NCH 3) 2], 3.2-2.8 (2H, m, H1, i 169 78094 H-6), 1.14 (3H, d, J 1 (_2, = 6.2, H- 2 '), 0.96 (3H, d, J 1 -CH 3 = 6.9, 1-CH-J. UV Λ pH 7.4: 296 πιμ (7809).

J max (5R, 6S) -paranitrobenzyl-6- (1R-hydroxyethyl) -4R-methyl) -3- [(1-methyl-1,2,3-triazol-4-yl) methylthio] -5,7-oxo -1-azabicyclo [3.2.o] hept-2-ene-2-carboxylate_ HO CH3 HQ jH3

10 ... cOpPNe Y

CO2PW 2} μ3 (5R, 6S) -paranitrobenzyl-6- (1R-hydroxyethyl) -3- (diphenylphosphono) -4R-methyl-7-oxo-1-azabicyclo [3.2.0] -15 hept-2-ene- To a cold (5 ° C) solution of 2-carboxylate (4.0 mmol) in acetonitrile (20 mL) was added 4-mercaptomethyl-1-methyl-1,2,3-triazole (735 mg, 5.7 mmol) and diisopropyl -ethylamine (646 mg, 5 mmol, 0.87 mL). After stirring at 5 ° C for 15 minutes and at room temperature for 1 hour, the reaction mixture was diluted with ethyl acetate (75 ml), washed with cold water, dried (MgSO 4) and evaporated. The residual foam was chromatographed on ultrapure Mushrooms (50 g); eluted with cold (5 ° C) ethyl acetate (200 ml) and then acetonitrile (200 ml). Relevant fractions 2 5 ...

The solutions were combined and evaporated to leave a foamy solid. Yield 1.1 g (58.1%).

IR (CH-C1-) V: 3605 (-OH), 1773 (β-lactam), 1710 * (es- & max teri). 1 HMR (CDCl 3) δ: 8.21 (2H, d, H aromatic), 7.63 (2H, d, Ho aromatic), 7.45 (1H, s, Ar-H), 5.34 (2H, ABq, • (n-benzyl), 4.40-3.80 (2H, m, H-1 ', H-5), 4.03 (2H, ABq, -SCH2-), 4.06 (3H , s, N-CH 3), 3.76 (1H, m, H1), 3.27 (1H, dd, J 5-6 = 2.5, Jx, _g = 6.5 Hz, H-6), 1.34 (3H, d, J1-2. = 6.1, H-2 '), 1.29 (3H, d, = 7.1, 1 "CH3).

78094 170

Example 36 (5R, 6S-3 - [(3,4-dimethylthiazolium-5-yl) methylthio] -6- (1'R-hydroxyethyl) -4R-methyl-7-oxo-1-azabicyclo [3.2 .03hept-2-ene-2-carboxylate (BMY-27,061) _ 5 Λ CF ^ O, CH, sK s CH3 [~ and A_sCH2H (^ -—> C "3 LTa" SCH2 "C> 3y-n2 γ, n 2) pa 2 \ 2y_ {1 ° 10 / V CH 2 N 2 CO 2 CO 2 CH 2 O 3 (10R, 6S) -allyl-6- (1'R-hydroxyethyl) -4R-methyl-3 - [(4- A solution of methylthiazol-5-yl) methylthio] -7-oxo-1-azabicyclo [3.2.O] hept-2-ene-2-carboxylate (1.15 g, 3.24 mmol) in dry dichloromethane (25 mL) was cooled. To 0 ° C and methyl trifluoromethanesulfonate (0.53 g, 3.24 mmol) was added dropwise (over 2 minutes) and the mixture was kept at 0 ° C for 40 minutes, after which triphenylphosphine (0.080 g) and tetrakis were added. (triphenylphosphine-2Q) palladium (0.080 g) followed by a 0.5 molar solution of potassium 2-ethylhexanoate (6.7 mL, 3.35 mmol) in ethyl acetate after maintaining the mixture at 0 ° C. minutes, allowed to warm to 22 ° C and add palladium comp Lex and triphenylphosphine (0.080 g each) were added three times every 2 minutes. The mixture was kept at 22 ° C for 3 hours, after which the dark reaction mixture was concentrated under reduced pressure and the residue partitioned between water (15 ml) and ether (15 ml). The organic phase was re-extracted with water (5 ml) and the combined aqueous phases were purified on a reverse phase silica gel-30 column (2.5 x 12 cm) using water followed by a mixture of water and acetonitrile (9: 1) as eluent. After a second chromatography, the title compound was obtained after lyophilization as a white powder: 0.248 g (21%); [α] D -9 ° (c 1.0, H 2 O); HPLC retention time: 4.0 minutes on a Merck 25 lichrosorb RP-18 4x250 mm column, eluting with 1 O-CH 2 CN 10%, flow rate 1 ml / min, UV detector at 294 nm; UV (H 2 O, pH 7.4 phosphate buffer) λ 258 (7160), 294 nm (8350); IR (KBr) V: c max ^, max 1740 (lactam C = 0), 1590 cm-1 (carboxylate C = 0); 1 HMR (D 2 O) δ: 1.05 (3H, d, J = 7.23, CH 3 -4), 1.11 (3H, d, J = 6.33, 171,78094 CH 2 -1 ') »2, 33 (3H, s, thiazoline CH 2 -4), 3.05-3.2 (1H, m, CH-4), 3.33 (1H, dd, Jh6> h5 = 2'82 'Jh6 / H1. = 6.0, H-6), 3.92 (3H, s, thiazolium CH 3 3 3), 3.94 (1H, dd, JR ^ Rg = 2.82, JH 5 Hi '= 6.67' H- 5) 4.07 (1H 'm' H_1,) '4'14 {2 *' ASq, JAB = δ 15.9, Λ \> = 52.3, SCH - 3), 9.55 ppm (1H , s, thiazole H-2, Δ -4 replaced by deuterium to KHCO 3). A 10 molar solution of compound in pH 7.4 phosphate buffer was found to have a half-life of 19 hours at 37 ° C.

Example 37 10 (5R, 6S) -3- [(1,3-dimethylimidazolium-4-yl) methylthio] -6- (1R-hydroxyethyl) -4R-methyl-7-oxo-1-azabicyclo lo [3.2.Pj hept-2-ene-2-carboxylate (BMY-27463) _ OH CH3 ch3 is L, (1,3-dimethylimidazol-4-yl) methanethiol trifluoromethanesulfonate_ 20 ru CH3 i 3 C «3 25 To a solution of (1-methylimidazol-5-yl) methylthioacetate (20.43 g, 0.12 ~ mol) in acetonitrile-ether (1: 2, 180 ml) was added dropwise (over 0.5 hour) methyl triflate (15 (84 mL, 0.14 mol) The mixture was stirred at 23 ° C under nitrogen for 1 h, then diluted with ether (500 mL) and the liquid was decanted to leave a red syrup which was washed twice with ether (100 mL). hydrochloric acid (200 ml, 6 N) was added and the resulting aqueous solution was heated at 65 ° C for 4 hours under an open oxygen atmosphere and evaporated, and residual ve-35 hydrochloric acid was removed by co-distillation with water to leave a yellow syrup; 2.4 g, 92%. A portion of the syrup was purified on a reverse phase column using water as eluent; IR (neat) ν ': 2550 (SH) max 172 78094 cm-1; 1 HMR (D 2 O) δ: 3.83 (s, CH 2 attached to the N atoms), 3.85 (s, CH 2 S), 7.34 (s, imidazolium H-5), 8.59 (s, imidazolium H-2). Analysis for C 7 H 11 N 2 O 3 S 2 F 3 · ®> 75H 2 O: Calculated: C 27.49; H 4.12; N 9.16; S 20.97; Found: C, 27.78; H 4.30; N 9.55; S 20.76.

(5R, 6S) -3 - [(1,3-dimethylimidazolium-4-yl) methyl] -6- (1R-hydroxyethyl) -4R-methyl-7-oxo-1-azabicyclo Π.3.2. Ojhept-2-ene-2-carboxylate_ 10 oh CH3 1- (PhO) 2POCl 2H JH3 ch3 A __- Jv 2 'N £ t (Hrv) 2. 10 ° * Pd / C. H. CAnAs / y H3C cryO 3-thiol Buffer, THF L-N | f

0 * ~ 4- N £ t (iPr) 2 ether δ J. ^ J

C00PNB CH3 15 (5R, 6S) -paranitrobenzyl 3,7-dioxo-6- (1R-hydroxyethyl) -4R-methyl-1-azabicyclo [3.2.0] heptane-2R-carboxylate (34.78 g, 0.096 mol) to a cold (-20 ° C) solution in dry acetonitrile (150 ml) kept under nitrogen was added diphenyl chlorophosphate (20.73 ml, 0.10 mol), diisopropylethylamine (17.42 ml, 0.10 mol) over 5 minutes and 4-dimethylaminopyridine (0.040 g).

The cooling bath was replaced with an ice water bath. The mixture was stirred at 0 ° C for 1 hour, cooled to -20 ° C and (1,3-dimethylimidazolium-4-yl) -methanethiol (32.4 g, 0.11 mol) was added sequentially. solution in acetonitrile (130 mL) and diisopropylethylamine (19.16 mL, 0.11 mol) at such a rate that the temperature remained between -15 ° C and -20 ° C (about 10 minutes). The cooling bath was replaced with 30 ice water baths. The mixture was stirred at 3 ° C for 1.5 hours, then diluted with water (840 mL). The resulting mixture was treated with Dowex 1x8 50 Cl (200 mL) and poured onto a reversed phase column (14 x 14 cm). A mixture of acetonitrile (25-35%) and water was used as the elution solvent. Fractions containing pure 35 carbapenem were passed through a Dowex 1x8 50 Cl column (4 x 58 cm) at 25 ml / min. Lyophilization of the aqueous solution gave a yellow solid, 27.6 g (55%); this was solubilized in buffer solution (KH 2 PO 4 - 173 78094

NaOH, 0.3-N, 615 mL, pH 7.0). After addition of 10% palladium-on-carbon catalyst (20 g), tetrahydrofuran (400 ml) and ether (600 ml), the mixture was hydrogenated at 23 ° C under 310 kPa for 2 hours and filtered through a pad of Celite. through. The organic phase was separated and extracted with water (2 x 50 mL). The aqueous phase was washed with ether (3 x 70 mL), placed under high vacuum to remove any traces of organic solvents, and poured into a reverse phase layer (14 x 13 cm). Eluted with a mixture of acetonitrile (0-4%) and water to give a pale yellow powder after lyophilization of the relevant fractions; 14.6 g, 78.8%; IR (KBr) ^: 1750 (β-lactam C = 0), 1590 (carboxylate)

1 η ITlclX

cm; 1 HMR (D 2 O) δ: 1.18 (d, J * 7.52 Hz, CH 3 attached to C-4), 1.29 (d, J = 6.35 Hz, CH 3 CHOH), 3.34 ( m, H-4), 3.47 (dd, 15 J = 2.75 Hz, J = 6.04 Hz, H-6), 3.84 and 3.87 (2s, CH 2 attached to the N atoms) ), 7.36 (s, imidazolium H-5), 8.64 (s, imidazolium H-2); UV (buffer, pH 7.0) λ: 298 (t 6932); 5-30 r max [α] 25 D -10.3 ° (c 0.34, H 2 O), t1 ^ 2 = 22.2 h measured at 36.8 ° C in buffer pH 7.4 at a concentration of 10 * M.

Claims (7)

78094 174 A process for the preparation of antibiotically active carbapenem derivatives of formula (I) 5 R 15 OH | JL \ CH-jCH --- S-A-f N © -R5 I 10. W - N - X N COOR 2 wherein R 5 is alkyl of 1 to 3 carbon atoms, carboxymethyl or allyl; n A is straight or side chain C 1 -C 6 alkylene; R is hydrogen, an anionic charge or a common easily removable carboxyl protecting group, provided that when R is hydrogen or a protecting group, a counterion is also present; R 2 is hydrogen or methyl; and _Q®_ 25 is a heteroaromatic group quaternized by R 1, which is a pyridinium, tetrazolium, imidazolium, triazolium, thiazolium or thiadiazolium group, and which may be substituted by 1 to 3 methyl, carboxymethyl or benzyl and is attached to the group A via a ring carbon atom, characterized in that a) the compound of formula (II) R 15 OH H 6 I - S - A - (- N II CH3CH - γ or N2coor2 '175 78094 wherein and A are as defined above and O is a heteroaromatic radical which is pyridyl, tetrazolyl, imidazolyl, triazolyl, thiazolyl or thiadiazolyl and which may be substituted by 1-3 methyl , carboxy-5-methyl or benzyl and attached to the group A via a ring carbon atom, and R is a common readily removable carboxyl protecting group, is reacted in an inert organic solvent with an alkylating agent of formula R 5 -X 'wherein R 5 is as defined above and X 'is a common leaving group such as halogen, so that the 15-O ring type of the substituent of the compound of formula II quaternizes with the group R ** to form the compound of formula (I1) R15? H - A - ^ S \ i® -R5 Χ, Θ 25 CH3CH - γ | I · J N - ^ 2 'CT ^ COOR ^ wherein R 1, R 2', A, 0 0 'R 2 and X' are as defined above; And, if desired, the counterion X 'is replaced by a different counterion and, if desired, the carboxyl protecting group 2 1 R is removed to give the desired unprotected compound of formula I or a pharmaceutically acceptable salt thereof, or b) (1) a compound of formula (III) 176 7 8 0 9 4 R15 OH HSI ΟΗ3Ϊη-- 5. I HI ^ N \ 2 'CT COOR ^ 1C, Λ I wherein RAJ and R ^ are as defined above, is reacted in an inert organic solvent with a reagent capable of coupling a conventional leaving group L such as a diphenylphosphoryloxy group to a compound of formula III to give a compound of formula (IV) 15 R15 OH H 8 I CH-iCH - Γ I (f IV J-N-V 20 O 2 COOR 2 'O I wherein L and R are as defined above; (2) reacting the obtained compound of formula IV in an inert organic solvent and in the presence of a base with a mercaptan reagent of formula hs-a-wherein A and as defined above, and modifying the obtained compound of formula II as described in a) above. the desired compound of formula I or a pharmaceutically acceptable salt thereof, or c) a compound of formula (IV) 177 7 8 0 9 4 0H H - 1 CH-aCH - 7 »- ^ 5 J 5 J — N-V 2. O 'COORz Process according to Claim 1, characterized in that R 1 is hydrogen. 2 I 1 c wherein R, RAJ and L are as defined above are reacted with a thiol compound of formula (VII) HS-A - ^ ^ yi®-R5 X® VII 15 wherein A and OR ^ are as defined above and X® is a counter anion, in an inert solvent and in the presence of a base, to form a carbapenem product of formula (I1) 20 r15 CH3CH-- S— A— ^ M © -R5 X® JN-2 ' CT1 COOR * wherein R1, R2, A, N, N®-R5 and X® are as defined above and, if desired, the carboxyl protecting group R2 'is removed to give the corresponding deprotected compound of formula I or a pharmaceutically acceptable salt thereof. Process according to Claim 1, characterized in that r 1 is methyl in the / i configuration. 178 78094