FI69845B - PROCEDURE FOR THE PREPARATION OF ANTIBACTERIAL ANTIBACTERIA - Google Patents

Description

Γ-i ^ n ....._ WHEN THE PUBLICATION IS.

B 11 UTLÄGGNINGSSKRIFT 6984 5 C (45) Patent granted '' p- 1 n *, 1 '1 - * rr o0 (51) Kv.ik // int.a.' C 07 D 499/00 FINLAND — FINLAND (21 ) Patent application - Patentansökning 793903 (22) Filing date - AnsÖkningsdag 1 3 · 1 2.79 (23) Starting date - Giltighctsdag 13.12 79

(41) Made public - Blivit offentlig ig Qg gQ

National Board of Patents and Registration Date of publication and publication. -

Patent and Registration Office in the United States of America and Public Utilities Publications 3i.i2.85 (32) (33) (31) Privilege claimed — Begärd priority 18.12.78 USA (US) 968663 (71) Bristol-Myers Company, 345 Park Avenue, New York, New York, USA (72) Marcel Menard, Canadiac, PQ, Alain Martel, Delson, PQ, Canada (CA) (74) Oy Koister Ab (54) Method for the preparation of new antibacterial penem derivatives - Förfarande för framstäl lning av nya antibakteriella penemderivat

The invention relates to a process for the preparation of novel antibacterial penem derivatives of the formula I and their pharmaceutically acceptable salts, Y -, -. s 1 1 κ I ', / (Alk) -A- (Alk') -NH-

L - f 2 I

Wherein "Alk is 2-alkylene, A is O, S or SO, Alk 'is C 2-6 alkylene and Y is hydrogen or lower alkyl which may be substituted by hydroxy.

69845 2

The formula for the penem ring system is 4 6 j 5! S.

- -4 Γ \ i I; \ - - 0 '7 1 \ 2 and its systematic name is 7-oxo-4-thia-1-azabicyclo / 3.2, ¢ 7-hept-2-ene. For simplicity, it is hereinafter referred to as "2-penem" and the numbering system used is as follows: 6j_5 ^, - S,! ; \ 2.

7 —N /; / 0 4 "^ <3

Various penem derivatives have been described in the prior art (see, e.g., DE-A-2,819,655, U.S. Patent No. 4,168,314, GB-A-2,013,674 and EP-A-0 003 892). However, the penem derivatives now described differ clearly from the known penem derivatives in terms of the 4-substituent of the azetidione ring.

Penem derivatives of formula I and their pharmaceutically acceptable salts can be prepared by reacting a compound of formula

O

Y- \ _S-C- (Alk) -A- (Alk 1) -N .. ti ri: - np \ ^ <Q> 3 i CO 2 R "where Y, Alk, A and Alk 'have the same meaning as above, R "is an easily cleavable ester-forming group and Q is phenyl or lower alkyl, cyclized in an inert organic solvent at a temperature slightly above room temperature and the reflux temperature of the solvent.

II

3,69845, after which the resulting compound is hydrogenated, and if desired, the resulting compound of formula I is converted into a pharmaceutically acceptable salt.

The compounds of the formula I can exist either as racemic mixtures (R, S-form) or as separate right- and left-turning (R- and S-form) optical isomers. Preferred are compounds having the 5-carbon atom configuration corresponding to natural penicillin (5R configuration). The substituents on the 5- and 6-positions of the 2,6-disubstituted penems may be in the cis or trans position relative to each other. The separation of the various optical and geometric isomers can be carried out by separation and mixing methods known per se.

Pharmaceutically acceptable salts include non-toxic carboxylic acid salts such as non-toxic metal salts, e.g. sodium, potassium, calcium, aluminum and magnesium salts, ammonium salt and salts of non-toxic amines such as N-dibenzylamines (triethylamine), procainamine, prochine. -phenethylamine, 1-ephenamine, N, N'-dibenzylethylenediamine, N-alkylpiperidine and other amines used to form salts of penicillins and cephalosporins. Pharmaceutically acceptable salts also include acid addition salts, e.g., salts with mineral acids such as hydrochloric, hydrobromic, hydroiodic, phosphoric or sulfuric acid, or with suitable organic carboxylic or sulfonic acids such as trifluoroacetic, p-toluenesulfonic, maleic, acetic with citric, oxalic, succinic, benzoic, tartaric, fumaric, almond, ascorbic and omic acids. The compounds of formula I may also be present in the form of internal salts, i.e. in the form of a zwitterion. The salts can be prepared by conventional methods for the preparation of salts of β-lactam antibiotics such as penicillins or cephalosporins.

The term "easily cleavable ester-forming group" has reached a certain meaning in β-lactam and peptide chemistry. Several such groups are known which are used to protect the carboxyl group in chemical reactions and which can subsequently be removed by standard methods to give the free carboxylic acid. Known ester protecting groups include 2,2,2-trichloroethyl, tertiary alkyl of 4 to 6 carbon atoms, tertiary alkenyl of 5 to 7 carbon atoms, tertiary alkynyl of 5 to 7 carbon atoms, alkoxymethyl, alkanoylmethyl of 2 to 7 carbon atoms, N-phthalimidomethyl, benzoylmethyl, halobenzoylmethyl, benzyl, p-nitrobenzyl, o-nitrobenzyl, benzhydryl, trityl, trimethylsilyl, triethylsilyl, b-trimethylsilylethyl, etc.

The choice of ester protecting group depends on the following reaction conditions for the group and the desired conditions for its removal. The expert is well able to select the appropriate group. The most preferred ester is p-nitrobenzyl ester, which is easily removed by catalytic hydrogenation.

Particularly preferred compounds of formula I are those wherein Y is hydrogen or β-hydroxyethyl, Alk is CH 2 / A is 0 and Alk is CH 2 CH 2.

The preparation of the compounds of formula I and the starting materials used can be described by the following reaction schemes.

Il 5

Reaction Scheme Ia 69845 V 0 1 s, _ ^ - 0 Ac \\ f XC-SNa Y-CH = CH-0Ac CSI ^ | - ^ - * J_ N pH 7.5 "O '' h O o v II y li _ ^ SC-X _ ^ ^ _ SC-X -> CHO I S0Cl2 V" 0 ^ -Νγ0Η C02R "0 n

V Π Y-v. CC Y

YV_.SC-X p0 - ^ SC-X Δ I —-> J-N p ^ Cl base A-N \ ^ P03 O ° co2r "co2r"

Y

^ __ Y g I - f \ v hydrogenation * Τ -, \ 1> -X __> I T V (Alk) -A- (Alk ·) -

Jr-N deprotection j___N *** 2

CO2R "° CO2H

__ g li AC = CH3C- 0 = c6h5- X = (Alk) -A- (Alk ·) -N3 6 69845

Reaction Scheme Ib 0 ^ S OAc ^ * Λ-CH-.C-SNa Y-CH = CH-OAc CSI „—---->

-> N

0 ^ H

Υ _ ^ _ ^ SAc _ \ YV_ ^ SAc - ^ CHO I SOCl2 0 ^ "N-H ^ R" 0 ^ “Ν ^ 0Η CO22"

Y

"V- ^ SAc P0 | -r" SAc MA

f -L-> -> J-N pj-01 base ^ -N \ ^ PO3 0 ° | co2r "co2r"

O

Y O v ^ '' Tr8 "x -“ - ®, VT * _i_ »0J — N ^ P03 ^ 0J — ΝΎ ^ Ρ03 C02Rm co2r" Y ^ r— - Γ \ _y hydrogenation ^ \ I // -> Γ | Alk) -A- (Alk ') -

X -— N '/' // NH

^ deprotection ^ -N 2 ° CO R "0 '

u 2 CO H

O 1

II

x-O-Θ = acylating agent MA = heavy metal salt X = (Alk) -A- (Alk ') - N3 li 7 69845

Reaction Scheme Ie Y - ^ OAc 0 CSNa Y-CH = CH-OAc CSI ^ —---> -> J_n eH 7 · 5

o ^ H

- <SC07 _-7> Υ'Κη-r ^ SC03 -> CHO I SOCl2 o ^ —nxh <V ο ^ ΝγΟΗ co2r "

Y \ -rSC0 P07 Y ^ 1-f5003 MA

Γ - ^ -> J-N -.Cl base ^ -N \ ^ PO3 0 ° CO2R "co2r"

y 0 v H

rrSM xg- ®, 'Vtsc'x o ^ -N ^ P03 * oJ_N ^ P03 C02R ”co2r" Y ^. s __l / - - [- f W, -A- „W) - s?' / deprotection i. _N _____> / 2 0 co_R ”o

2 CO2H

X = (Alk) -A- (Alk ') - N3 8 69845

In Reaction Schemes Ia, Ib and Ie, a vinyl ester containing the desired 6-substituent (Y = H or a radical as defined for Compounds I) is converted to the desired 1-substituted 4-acetoxy-2-azetidinone by cycloadditioning chlorosulfonyl isocyanate (CSI) and then reducing with an organic reducing agent. sodium sulfite. More conveniently, the CSI reaction is performed in an inert organic solvent such as diethyl ether at a temperature of 0 ° C or below. The reduction step can be performed in an aqueous or aqueous-organic reaction mixture at a temperature of 0 ° C or below at a slightly basic pH.

According to Reaction Scheme Ia, azetidinone is reacted with thiolic acid 0

II

X-C-SH or a salt thereof in a suitable solvent (e.g. water or an aqueous organic solvent). The displacement reaction is preferably carried out at or below room temperature at a slightly basic pH (about 7.5). When Y is not H, the cis and trans isomers of the azetidinone formed are preferably separated at this stage of the process (e.g. by chromatography). In Reaction Schemes Ib and Ie, 4-acetoxy-2-azetidinone is converted to 4-acetylthio-2-azetidinone and 4-tritylthio-2-azetidinone products, respectively, by nucleophilic substitution with thioacetic acid or triphenylmethyl mercaptan (or a salt thereof such as sodium salt), respectively.

Next, 4-thioazetidinone is reacted with glyoxylate ester 0

II

HC-CC 2 R ", where R" is an easily removable ester protecting group, or a reactive oxo derivative such as a hydrate in an inert organic solvent (e.g. benzene, toluene, xylene, etc.) and preferably at elevated temperature (e.g. 50 ° C: and at a maximum return temperature). When an ester hydrate is used, the water formed can be removed azeotropically or by molecular sieves. The hydroxy ester product is formed as an epimer mixture which, if desired, can be purified by chromatography or used directly in the next step.

69845

Conversion of the hydroxy ester to the corresponding chloroester is accomplished by reacting a chlorinating reagent (e.g., SOCl 2, POCl 3, PCI, etc.) in an inert organic solvent (e.g., tetrahydrofuran, diethyl ether, methylene chloride, dioxane, etc.) with a basic, preferably aliphatic, tertiary. in the presence of a triethylamine) or a heterocyclic tertiary amine (e.g. pyridine or collidine). The reaction is preferably carried out at a temperature between about -10 ° C and room temperature. The chloroester product is obtained as an epimer mixture which, if desired, can be purified before use in the next step.

The phosphorane intermediate can be obtained by reacting the chloroester with a suitable phosphine (preferably triphenylphosphine or trialempkylphosphine such as tri-n-butylphosphine) in an inert organic solvent such as dimethylformamide, dimethylsulfate, dimethyl sulfoxide, tetrahydrofuran, tetrahydrofuran, dimethoxyethane, dioxane, dioxane. in the presence of a base, preferably an organic tertiary amine such as triethylamine, pyridine or 2,6-lutidine. The reaction is preferably carried out at a temperature between room temperature and the reflux temperature of the solvent system.

In Reaction Scheme Ia (where the 2-substituent is already attached), the phosphorane intermediate is converted to the desired penem by thermal cyclization in an inert organic solvent at a temperature slightly above room temperature and the reflux temperature of the solvent system. Most conveniently, the cyclization is performed under reflux. Suitable inert organic solvents include aliphatic, cycloaliphatic or aromatic hydrocarbons (e.g. benzene, toluene, hexane, cyclohexane), halogenated hydrocarbons (e.g. methylene chloride, chloroform, carbon tetrachloride), ethers (e.g. diethyl ether), ethers (e.g. , dimethoxyethane), carboxylic acid amides (e.g. dimethylformamide), di-C 1 -C 4 alkyl sulfoxides (e.g. dimethyl sulfoxide) or C 1 -C 6 alkanol (e.g. methanol, ethanol, tert-butanol) or mixtures thereof.

10 69845

In Reaction Schemes Ib and Ie, phosphorane is converted to a heavy metal mercaptide of the formula "rs f"

] 1 | M III

4 - N \ ^ P (Q) 3! 0 1 i CO R "J x U— 2.

or, SH is C 1 (2) 3 CO R "wherein Q is phenyl or lower alkyl, x is 1 or 2 and M is Cu (II), Pb (II) or Hg (II) when x is 2, or Ag (I) when x is 1. The mercaptide is formed by reacting phosphorane with a salt of Hg (II), Pb (II), Cu (II) or Ag (I) or (methoxycarbonyl) mercury (II) with acetate in a methanolic solvent and with an organic or inorganic base such as aniline, pyridine, collidine, 2,6-lutidine, alkali metal carbonate, etc. The preferred base is pyridine The reaction may be carried out at room temperature or, if desired, under moderate cooling or heating. the anion (A) may be any anion which gives a soluble salt in the chosen solvent, e.g. able to connect

II

11 69845 0 0 o tl tl It part X-C-. The acylating agent (XC-Φ) may be an acid XC-® or a reactive functional derivative thereof such as an acid halide (preferably an acid chloride), an acid azide, an acid anhydride, a mixed acid anhydride, an active ester, an active thioester, etc. The acylation is performed in an inert solvent. such as methylene chloride or an ether such as dioxane, tetrahydrofuran or diethyl ether) and using an acid derivative in the presence of an acid acceptor such as tri-lower alkylamine (e.g. triethylamine) or a tertiary organic base such as pyridine, collidine or 2,6-lutidine. When free acid is used, the acylation is carried out in the presence of a suitable condensing agent, e.g. a carbodiimide such as N, N1-dicyclohexylcarbodiimide. The acylation of mercaptide can be performed over a wide temperature range, but -20 ° to + 25 ° C is recommended. The phosphorane formed after acylation is cyclized as described above to give the desired penemester.

It has been found that the preparation of phosphorane via the mercaptide intermediate (Reaction Schemes Ib and Ie) results in a much purer product than using the more conventional route described in Reaction Scheme Ia.

The hydrogenation in the last step can be carried out in the presence of a noble metal catalyst such as palladium or rhodium and also their derivatives such as oxides, hydroxides or halides. If desired, said catalyst is on a conventional support such as carbon or diatomaceous earth. A non-reducing inert aqueous or non-aqueous solvent such as water, ethanol, methanol, ethyl acetate, tetrahydrofuran, diethyl ether or dioxane is used. The hydrogenation can be carried out at normal or elevated pressure and suitably at room temperature for about 1 to 5 hours depending on the solvent and the catalyst. When an equivalent weight amount of a base such as an early metal or crude alkali metal hydroxide or amine is used in the hydrogenation, the product can be isolated in the form of a carboxylic acid salt.

69845

Reaction Scheme II

SC07 I ...... "f" 0 XH 0 \

B

Y

Hj, ___ SC0

_ * ^! I

emas! '

L_N

\ B

deprotection N. MA / base (/ ^

Y> sc0, "0_____SM

I - 0 *

(Γ \ OB

0 CHO X-C-0 CO-jR "v

II

13 69845 o Y '^. SC0-, ____ sc-x I!

, - N j OH J-N

N, ^ o B

C02R "deprotection SOC1 4 * 0 Y SC0t Y: sc-x> -! J -

C1 N

<y 0 \

B

CO 2 R "

P0 CHO

T

base CO-R "Ψ /

V

w 0

Λ ^ SC0 ^, y II

--- ^ J ^ ^ .sc-x

J P <^ 3 / - N OH

s' yO \ ~ l 0 0 ^ \ vA /> 'C02R' C02R "MÄ / base S0C1 V φ 14 69845 SM γ π - [^ SM ^ sc-x oJ-N- ^ PO3 oJZ] Cl

O

11 ^ P0, V / x-c- Θ 3 v. base

° V O

Y-V ςρ γ Y II

_ ^ SC-X S_ ^ sc-x O J — N \ ^ P03 oJ— 'c02R "C02R"' ΡΪ co2R "C02R" hydrogenation deprotection hydrogenation deprotection Ψ V / 0 o N ^ \

C ° 2H CO 2 H

B = ring nitrogen blocking group X = (Alk) -A- (Alk ') - N3

II

15 69845

According to Reaction Scheme II, the ring nitrogen of 4-tritylthio-2-azetidinone is protected with a conventional, easily removable blocking group such as triorganosilyl (e.g., trimethylsilyl or tert-butyldimethylsilyl), methoxymethyl, methoxyethoxymethyl, etc., halide, methoxyethoxymethyl the position is performed by reacting a suitable electrophile with an N-protected azetidinone in the presence of a strong base. At this point, the method branches into two pathways depending on the duration of azetidinone deprivation.

In the second route, the N-protected intermediate is deprotected by conventional methods (e.g., acid hydrolysis) and then converted to 2,6-penem via esterification, chlorinated to hydroxyester, converted to chloroester to phosphorane, converted to phosphorane to heavy metal mercaptide, acylated to the mercaptide, phosphorane to give the 2,6-penemester and removal of the carboxyl protecting group.

The steps of the alternative route are the conversion of the N-protected azetidinone to a heavy metal mercaptide, the acylation of the mercaptide O

II

with X-C-®, deprotection of N, deprotection of azetidinone and glyoxylate ester, chlorination, reaction of chloroester and phosphine to phosphorane, cyclization of phosphorane to penem ester, and deprotection of the carboxyl to give 2,6-penem. The reaction conditions of these steps are the same as above.

In the preparation of 2-penem or 2,6-penem compounds according to the above methods, the free functional groups of the substituent X or Y which do not participate in the reaction can be temporarily protected in a manner known per se, e.g. by etherification or esterification, mercapto groups by esterification and free carboxyl or sulpho groups, e.g. by esterification and also by silylation. Upon completion of the reaction, these groups may, if desired, be liberated individually or together in a manner known per se.

16 69845

The penem derivatives of formula I and their pharmaceutically acceptable salts have been found to be potent, broad-spectrum antibacterial agents useful in the treatment of infectious diseases caused by both gram-negative and gram-positive organisms in animals and humans.

The compounds of formula I were subjected to in vitro antibiotic screening for various microorganisms. After dissolving the samples in water and diluting with the nutrient solution, they were found to have the following minimum inhibitory concentrations (MEK) micrograms / ml for said microorganisms as determined by incubation overnight at 37 ° by the tube dilution method.

MEK micrograms / ml

Compound (Example No.) Organism 1_ 2 ^ 3 4_

Streptococcus pneumoniae A9585 .03 .13 .13 .5

Streptococcus pyogenes A9604 .03 .13 .13 .5

Staphylococcus aureus A9537 .06 .13 .13 .5

Staph aureus +50% serum A9537 .5 1 .5 4

Staphylococcus aureus A9606 .25 32 16 4

Staphylococcus aureus A15097 1 125 16 63

Streptococcus faecalis A20688 4 32 63> 63

Escherichia coli A15119 .5248

Escherichia coli A20341-1 .5 8 16 32

Klebsiella pneumoniae A15130 2 4 8 32

Klebsiella species A204 68 2> 125> 63> 63

Si 17 69845 MEK micrograms / ml

Compound (Example No.) Organism 1 2

Proteus mirabilis A9900 1 2 4 8

Proteus vulgaris A21559 1 8 16 16

Proteus morganii A15153 4 8 8 16

Proteus rettgeri A21203 2 2 8 16

Serratia marcescens A20019 2 2 8 32

Enterobacter cloacae A9659 8 8 32 32

Enterobacter cloacae A9656 2 4 32 63

Pseudomonas aeruginosa A9843A 16 63 63 63

Pseudomonas aeruginosa A21213 8 125> 63 63

Hemophilus influenzae A9833 - - -

Haemophilus influenzae A21522 - - -

Bacteroides fragilis A20931 - - -

Bacteroides fragilis A20929 - - -

The novel compounds may be formulated into pharmaceutical compositions which contain, in addition to the active ingredient, a pharmaceutically acceptable carrier or diluent. The compounds can be administered both orally and parenterally. The pharmaceutical preparations may be in solid form, e.g. capsules, 69845 18 tablets or granules, or in liquid form, e.g. solutions, suspensions or emulsions. In the treatment of bacterial infectious diseases in humans, the compounds of the formula I can be administered orally or parenterally in an amount of about 5 to 200 mg / kg / day and preferably about 5 to 20 mg / kg per day in separate doses, e.g. three or four times daily. They are administered in dosage units containing, for example, 125, 250 or 500 mg of active ingredient in association with suitable physiologically acceptable carriers or diluents.

The following are illustrative examples of the invention. All temperatures are in degrees Celsius. In the examples, e.g. the following abbreviations: CSI chlorosulfonyl isocyanate pet.ether petroleum ether b.p. boiling point NMR nuclear magnetic resonance spectroscopy h hour ether diethyl ether (unless otherwise stated)

Trademarks of Celite Johns-Manville Products Corporation PNB p-nitrobenzyl sp. melting point LAH lithium aluminum hydride n-BuLi n-butyllithium MIBK methyl isobutyl ketone

Et C2H5-

Tr -C <C6Hs> 3

Me CH3 THF tetrahydrofuran

Ph phenyl DMF dimethylformamide TEA triethylamine PNBG p-nitrobenzylglyoxylate THP tetrahydropyranyl TFA trifluoroacetic acid HMPT hexamethylphosphoric triamide (or HMPA) LDA lithium diisopropylamide

Ac CH3CO-

Ms CH 3 SO 2 - DMAP 4-dimethylaminopyridine

II

19 τ * μ- η Ί ·, 69845 Manufacture of starting materials

Example I

1- (p-Nitrobenzyloxycarbonylmethyltriphenylphosphoranyl) -4- (silver mercaptidyl) -2-azetidinone ______-SAg 0J ~

'STT

0AC CD2PNB _ ^ rf ^ Λ ί _ '# N, NaOMe 0 H 2 0 “

A suspension of 13.8 g (0.04 mmol) of triphenylmethyl mercaptan in 90 ml of methanol was purged of the gases for half an hour with a stream of nitrogen. The mixture was cooled to 0 ° and 2.4 g (0.5 mol, 50% oil dispersion) of sodium hydride was added portionwise. The resulting solution was stirred for five minutes and 7.7 g (0.059 mol) of 4-acetoxyazetidinone in 55 ml of water were added rapidly. 4-Triphenylmethyl mercaptoazetidinone (2) precipitated immediately. The mixture was stirred for four hours at room temperature. The solid was isolated by filtration, washed with water and dissolved in methylene chloride. The methylene chloride solution was washed with dilute hydrochloric acid, water, sodium bicarbonate solution, water and brine and dried over magnesium sulfate (89.8%, m.p. 146.5 ° -147.5 °).

Analysis for C 22 H 19 NOS calculated: C 76.49 H 5.54 N 4.05 S 9.28 found: C 7.54 H 5.60 N 4.00 S 9.36. 1 H (ppm, CDCl 3): 7.60-7.10 (15H, m, H-trityl), 4.62 (1H, broad, NH), 4.40 (1H, dd, J 4 -3 trans = 3'0 'J4-3 cis = 5' H-4> '3'24 (1H' ddd 'Jgem = 15' J3-4 cis "5 'J3-NH = 1'8' H-3) '2'81 <1H 'ddd> Jgem' 15 'J3-4 trans = 3'0' J3-NH '1'2' «-3> -V co <chc13>! 1760 '^ hH 3340 · STr P ° s_ ^ STr r — f co2pnb j-fSTr I j A, *.-Lv ”'Av

2 C02PNB 3 C02PNB

20,684 5 4.54 g (0.02 moles) of hydrated p-nitrobenzylglyoxylate and 6.90 g (0.02 moles) of azetidinone were heated under reflux in 24 benzene through a Dean-Stark condenser filled with a 3 A molecular sieve. . An additional 2 x 454 mg (2 mmol) of glyoxylate was added and heated under reflux for 18 h after each addition. The mixture was diluted with ether, washed with 5% aqueous hydrochloric acid, water, 5% aqueous sodium bicarbonate, water and brine. Dried over magnesium sulfate (12 g, quantitatively. A small portion of the epimer mixture was separated on a silica gel plate (CH 2 Cl 2: 6: 4). Isomer A:

Rf = 0.87, m.p. 170.5 ° -171.5 °, J (ppm CDCl 3): 8.07 (2H, d, J = 9, Hm aroma), 7.45 (part of d, Ho aroma), 7.40-7 .00 (L5H, m, trityl), 5.25 (2H, s, CH2-PNB), 4.75 (1H, s, HCO), 4.37 (1H, dd, J3_4 trans = 3, J3_4 cis = 4, H-3), 2.83 (1H, dd, J = 16, J. O. = 4, H-4), 2.10 (1H, dd, J = 16, J.-.

gem 4-3 cis (gem 4-3 trans 3, H-4), 1.42 (broad), OH). y> c = 0 (CHCl 3): 1760 (shoulder), 1525 'V'OH 3475 * 2

Isomer B:

Rf = 0.75, m.p. 152 ° -153 °, / (ppm, CDCl 3): 8.13 (2H, d, J = 9, Hm aroma), 7.47 (2H, d, J = 9 / Ho aroma), 7.40-7 .00 (15H, m, trityl, 5.30 (3H, s, CH 2 -PNB, HCO), 4.45 (1H, t, J = 3.5, H-4), 2.90-2.70 (2H, AB part of ABX, H-4), 1.55 (broad, 0H) '^ 0 = 0 <CHCl3): 767' 1755 (shoulder'Nno2 15 25, 0H 3500.

STr _ ^ STr soci j — r j -> _ C1: n ^ 0H pyridine / \, ·

0 I C02PNB

C02PNB

3 4 Under a nitrogen blanket, 12 g (21.7 mmol) of azetidinone 21 69845 were treated in 150 ml of cold (-15 °) THF (molecular sieve dried) with 1.9 g (24.1 mmol, 1.94 ml) of pyridine. and dropwise with 2.86 g (24 mmol, 1.88 mL) of thionyl chloride. The mixture was stirred for 45 minutes at -15 °. The precipitate was isolated by filtration and washed with benzene. Evaporation of the solvent gave a residue which was dissolved in benzene and treated with 11.7 g of activated carbon (94%, crystallized from chloroform), δ (ppm, CDCl 3): 8.17 (2H, d, J = 8,

Hm aromatic), 7.67-7.00 (17H, m, Ho aromatic »Tr-H), 5.80 (s, HC-Cl), 5.37, 5.33 (2s, HC-Cl, CH (PNB), 4.81 (1H, m, H-4), 3.27-2.40 (2H, m, H-3),>) c = Q (KBr film): 1785, 1770, NQ 1525.

STr STr, L N, C1 03P_v. J- N. P0 0 ^ 2,6-lutidine / | ' co2pnb co2pnb 4 5

A solution of 11.6 g (20.2 mmol) of chlorazetidinone in 4-100 mL of THF (distilled over LAH) was treated with 7.86 g (30.0 mmol) of triphenylphosphine and 2.36 g with (2.56 mL, 22.0 mmol) 2,6-lutidine. The mixture was heated under reflux for 72 h. The precipitate was isolated by filtration and washed with ether. The organic solution was washed with 2% aqueous hydrochloric acid and 5% aqueous bicarbonate and dried over magnesium sulfate. Evaporation of the solvent gave a residue which was purified on a 200 g layer of silica gel. The desired phosphorane was eluted with 30, 40 and 50% ether-benzene (11.4 g, 70.4%, mp 201 ° -202 °).

Analysis for C 49 H 40 N 2 O 5 SP: Calculated: C 73.57 H 5.04 N 3.50 S 4.01, Found: C 73.58 H 4.91 N 3.44 S 3.87.

>) (CHCl 3): 1740, phosphorane (1620, 1610), 1525.

c-o 3 _ ^ STr AgNO_, SAg I ^ —-—> i Γ 0> -γ »3 Pyridiln ^ tfJ_N ^ P03

C02PNB C02PNB

_5 6 22 6 9 8 4 5

1.6 g (2 mmol) of 4-trityl mercaptoazetidinone were dissolved in 5-20 ml of CH 2 Cl 2 and the solvent was returned at 55 ° -60 °.

Phosphorane at 5-55 ° -60 ° was dissolved in 32 parts of preheated (55 ° -60 °) methanol. Immediately after obtaining a solution of 5 methanol, it was treated with a preheated (55 ° -60 °) mixture of 16 ml of 0.15 m (1.2 equivalents) of silver nitrate in methanol and 174 mg (178 μΐ, 2.2 mmol) of , 1.1 equivalents) pyridine, the heating bath was immediately removed and the mixture was stirred for 2 h at room temperature and 1 h at 0. Silver mercaptide (> isolated by filtration, washed twice with cold (0 °) methanol and three times with ether, (1.12 g, 84.5%, mp 130 ° -135 °, dec.).

^ c = o (CHCl 3): 1795, 1725 (shoulder), phosphorane (1920, 1605), 0no2 1530 '

Example II

1- (p-Nitrobenzyloxycarbonylmethyltriphenylphosphoranyl) -4- (silver mercaptidyl) -2-azetidinone SCOCH βΑ <3/3 K, CQVA ^ °, _ (-f

_ N Me OH J — N

ο '^^, -. ΡΓΗ 0' -ypfhj

COOPUa COOPNB

7 i

A solution of 1.796 g (3.0 mmol) of phosphorane in 1 to 3 mL of chloroform was diluted with 90 mL of methanol, cooled to 0 ° under a nitrogen blanket, and treated sequentially with 0.51 g (3.0 mmol) of silver nitrate and With 33 g (2.4 mmol) of potassium carbonate. The reaction mixture (protected from light) was stirred for 15 minutes at 0 °, the cooling bath was removed and stirring was continued for 3 h. The reaction mixture was cooled to -10 °, stirred for 1 h, filtered, the silver mercaptide was washed successively with cold methanol and ether to give 1.91 g, m.p. . 138 ° -145 °, dec., 96%.

IR (nujol) cm -1: 1748, 1620 and 1605. Analytical sample 23,69845 was obtained

by preparative TLC (ethyl acetate), m.p. 140 ° -145 ° decomposes. Analysis for C 54 H 21 N 2 O 2 SPAg: 1 * step C 54.31 H 3.65 N 4.22 S

4.83 Found: C 54.11 H 3.48 N 3.92 S 4.62.

Example III

1- (p-Nitrobenzyloxycarbonylmethyltriphenylphosphoranyl) -4- (silver mercaptidyl) -2-azetidinone A. Use of aniline as a base _ ^ SCOCH3 AgNO3 -j ^ Ag 0 'C = PPh MeOHH θ' ^ C = PPh I 3 | 3

COOPNB COOPNB

7

A solution of 1.8 g (3.0 mmol) of phosphorane 7 in 4 mL of chloroform was diluted with 90 mL of methanol, cooled to -15 ° under nitrogen and treated sequentially with 0.56 g (3.3 mmol) of chloroform. ) silver nitrate and 1.5 ml (16.5 mmol) of aniline. The reaction mixture (protected from light) was stirred at -15 ° for half an hour, then the cooling bath was removed and stirring was continued for 24 h. The reaction mixture was cooled to -10 °, stirred for 1 h and filtered. The silver mercaptide was washed successively with cold methanol and ether to give 1.55 g, m.p. 114 ° -115 °, dec., 77.9%.

IR (nujol) cm same as in Example II.

Silver 1- (p-nitrobenzyl-22-triphenylphosphoranylidene-2'-acetate) -2-azetidinone-4-thiolate B. Use of 4-dimethylaminopyridine (DMAP) as a base - ^ ° CH3 AgNO3 / DMAP_ ^ SAg P4> 3 CH2C12 / CH3OH "O iN YP <^ 3 CO2PNB co2pnb η λ 69845

A solution of 17.96 g (30 mmol) of the above S-acetylphosphorane in 450 ml of 1: 2-methanol-dichloromethane was purged with nitrogen (5-10 minutes), cooled to 5 ° and treated successively with 5.35 g: (31.5 mmol) of silver nitrate and 3.85 g (31.5 mmol) of 4-dimethylaminopyridine. The ice bath was removed and the solution was heated vigorously for 2 h and then stirred at room temperature for 1 h. The colored reaction mixture was treated with charcoal, filtered and evaporated. The residue was redissolved in a minimum of dichloromethane and added dropwise with stirring to 300 ml of cold methanol. The precipitated silver salt was isolated by filtration, washed with ether, dried to give 18.1 g (91%), IR (CHCl 3) ν max: 1745 (β-lactam C = O) and 1607 cm -1 (ester C = 0).

Silver 1- (p-nitrobenzyl-21-triphenylphosphoranylidene-2 "-acetate) -2-azetidinone-4-thiolate C. Use of diazabicyclounaquene (DBU) as base _ SCOCH3 AgN03 ^ SAg 0 Y P03 DBU 'Me0H 0 Νγ 3

C02PNB C02PNB

36.0 g (0.060 mol) of the above S-acetylphosphorane were dissolved in 120 ml of methylene chloride. The solvent was evaporated to give an oil. This oily residue was dissolved in 240 ml of warm (35 °) methanol and treated rapidly with 10.68 g (0.0628 moles) of silver nitrate dissolved in 420 ml of methanol. The resulting solution (or suspension) was stirred for 5 minutes at room temperature, cooled (ice bath) and a solution of 8.96 ml (0.060 mol) of DBU in 20 ml of methanol was added over 5 minutes. The mixture was stirred for five minutes. The solid was isolated by filtration, washed once with cold (0 °) methanol and ether, dried in vacuo to give 37.0 g (93%), IR (nujol) λmax: (C = O) and 1600 cm-1 (phosphorane).

25 69845 D. Use of pyrrolidine as a base

Silver 1- (p-nitrobenzyl 2'-triphenylphosphoranylidene-2-acetate) -2-azetidinone-4-thiolate

SCOCH

—Y pyrrolidine _ ^ SAg „AgN0o I!

NxC-PPh_ I 3 C-PPh_ I i

COOPNB

COOPNB

To a cold (0 °) solution of 0.60 g (1.0 mmol) of 4-acetylthio-1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2 '' - acetate) -2-azetidinone in 2 mL of CU ClCl 2, 4 mL of MeOH, 7.86 mL of 0.14 M MeOH in silver nitrate (1.1 mmol) and a solution of 0.92 mL (1.1 mmol) of pyrrolidine in 2 mL were added. The cooling bath was removed, the reaction mixture was stirred for 1.75 h, cooled to -10 °, stirred for 0.25 h and filtered The solid was washed with cold MeOH, dried in vacuo to give 0.548 g, mp 115 °, 82.4% IR (nujol) λ max 1755 (C = O) and 1600 cm -1 (aromatics).

Example IV

1- (tert-butyldimethylsilyl) -4-tritylthio-2-azetidinone-1-rSO3 <r'-N h "</ o,»;

1.62 ml (11.6 mmol) of triethylamine were added dropwise over 5 minutes to a cooled (0 °) and stirred solution of 3.5 g (10.1 mmol) of 4-tritylthio-2-azetidinone and 1.68 g ( 12.7 mmol) of chloro-tert-butyldimethylsilane in 35 ml of DMF. The reaction mixture was stirred at room temperature for 18 h and diluted with 250 mL of water and 200 mL of ether. The organic phase was washed with 3 x 50 mL of water, dried, concentrated to give 4.33 g of an oil. Crystallization from pentane gave a total of 4.1 g (89%) of the title compound as a white solid, m.p. 113 ° -114 °. Δ (ppm, CDCl 3): 7.45 (15H, m, aromatics), 4.2 (1H, dd, H-4), 2.63 (1H, dd, J = 4, J = 16, H- 3), 2.13 (1H, dd, J = 2, J = 16, H-3), 1.0 (9H, s, t-Bu), 0.35 (6H, s, Me). V c = 0 1735 Analysis for C 12 H 12 NOSSi: calculated: C, 73.15, H 7.24 N 3.05 S 6.97%, found: C, 73.27, H 7.32 N 2.97 S 6.94%.

Example V

1- (tert-butyldimethylsilyl) -3- (1'-hydroxy-11-ethyl) -4-tritylthio-2-azetidinone (4 isomers)

OH

SC0. sc03 a) --f '3 ^ Cii3 tBu I - N \. s. .-

<^ - N t-BU O

0 '"sr" (He) 2

Me) 2

3.4 ml of 1,6-m-n-butyllithium (5.44 mmol) in 0.847 ml (6.23 mmol) of diisopropylamine in 30 ml of THF were added over 5 minutes. kept at -78 °. After half an hour a solution of 2.0 g (4.4 mmol) of 1- (t-butyldimethylsilyl) -4-tritylthio-2-azetidinone in 20 ml of THF. After 15 minutes, 10 ml of acetaldehyde were added in one portion and after a further 15 minutes, 100 ml of water. The mixture was acidified (pH 5-6) with dilute hydrochloric acid and extracted with 3 x 30 mL of ethyl acetate. The organic phases were washed with brine, dried, concentrated to give an oil which by TLC consisted of a mixture of four isomers (named isomers A, B, C and D in descending polarity order).

27 6 9 8 4 5

Crystallization of the oily residue from ethyl acetate-pentane gave isomers B and C as a white solid and A and D were in mother liquors. The four pure compounds were obtained by preparative chromatography (Waters 500) of the above solid and mother liquors. The relative proportions were 17% A, 32% B, 39% C and 12% D. When in the reaction described above the THF was replaced with ether and the reaction was stopped after one minute at -78 ° for A, B, C and D The relative shares of were 12.9, 30.5, 38.2 and 18.4%. When the reaction was allowed to reach ether at 20 ° in two hours before stopping, the proportions were 13.4, 24.6, 44 and 18%. When one molar equivalent of anhydrous magnesium bromide was added to the reaction mixture, the ratios changed to 19.2, 19.7, 30.1 and 31%.

Isomer A: This isomer has cis-stereochemistry at C 1 -C 4. It is a racemic mixture of (1 ', S, 3R, 4R) and (1'R, 3S, 3S) enantiomers. The compounds which are subsequently derived from compound A are referred to as "isomer A". They consist of an enantiomeric mixture and have the same configuration at and.

Compounds derived from Compound A by a reverse reaction of the configuration are referred to as "isomer D" if the translation occurs at C 1, and as "isomer C" when the translation occurs at C 3, m.p. 152 ° -153 °, 1 HMR (CDCl 3): δ 8.0-6.8 (15H, m, aromatics), 4.30 (1H, d, J = 5.5, H-4), 3, 78 (1H, m, H-1 '), 3.10 (1H, dd, J = 5.5, J = 10, H-3), 1.22 (3H, d, J = 6.5, CH 3 >, 0.95 (9H, s, Bu), 0.27 (6H, 2s, CH 3) Analysis for C 75 H 24 N 2 O 2 Si: calculated: C 75.52 H 7.40 N 2.78 S 6, 36%, found: C 71.28 H 7.41 N 2.48 S 6.19%.

Isomer B: This isomer has trans stereochemistry at C3-C4. It is a racemic mixture of (11, R, 3S, 4R) and (1'S, 3R, 4S) enantiomers. Compounds having the same configuration at C 1, C 3 and C 4 are referred to as “isomer B.” IR (CHCl 3, max: 1745 -1 (C = O), mp 158-159 °, 1 HMR (CDCl 3): ε 7.60-7.10 (15H, m. Aromatics), 4.02 (1H, d, J = 0.8, H-4), 3.32 (1H, dd, J = 3.0, J = 0.8, H-3), 3.55-3.15 (1H, m, H-1 '), 0.88 (12H, CH 3, t-Bu), 0.16 (6H, s, CH 3) .

Isomer C; This isomer has trans stereochemistry at C3-C4 · It is a racemate formed by the (1'S, 3S, 4R) and (11R, 3R, 4S) enantiomers 28 69845. Compounds having the same configuration at and referred to as "isomer C", m.p. 134-136 °, 1 HMR (CDCl 3). 7.60-7.10 (15H, m, aromatics), 4.32 (1H, d, J = 1.8, H-4), 3.02 (1H, dd, J = 2.7, J = 1.8, H-3), 3.0-2.5 (1H, dg, J = 2.7, J = 6 H-1 '), 1.02 (3H, d, J - 6, CH ), 0.95 (9H, s, t-Bu), 0.27 (6H, s, CH 3), IR (CHCl 3) ν max: 1735 cm -1 (C = O).

Isomer D; This isomer has cis stereochemistry at C3-C4. It is a racemate of the (1 ', R, 3R, 4R) and (1'S, 3S, 4S3 anantiomers. Compounds having the same configuration at C, C, and C are termed "Isomer D", mp 171 ° -172 °, η X ό *> HMR (CDCl 3). ^ 7.80-6.90 (15H, m, aromatics), 4.70 (1H, d, J = 4.5, H-4), 3.02 (1H, dd, J = 4.5, J = 0.5, H-3), 2.39 (1H, dg, J = 0.5, J = 6.5, H-1 ') , 1.0 (3H, d, J = 6.5, CH 3), 0.97 (9H, s, t-Bu), 0.32 (6H, s, CH 3). : Calculated: C 71.52 H 7.40 N 2.78 S 6.36%, Found: C 71.27 H 7.43 N 2.51 S 6.31%.

OH

A,: SC0 / SC ^ 3

, —K J 7 — I

BH! I

b) // ~ V ^ ^ ° S \ \

A solution of 1.0 g (2 mmol) of trans-3-acetyl- (tert-butyldimethylsilyl) -4-trityl-2-azetidinone in 30 mL of THF was added dropwise to a cooled (0 °) and stirred nitrogen blanket. to a suspension of 0.38 g (10 mmol) of sodium borohydride in 120 mL of THF. The ice bath was removed and the mixture was stirred at room temperature for 4 h. It was poured into ice-cold 1N hydrochloric acid (pH 6), stirred for 15 minutes and extracted three times with ether. The combined ether extracts were dried, concentrated to give 1.04 g of an oil which was crystallized from pentane and

II

29 6 9 8 4 5 gave the title compounds as a 70:30 mixture of C and B isomers, m.p. 1190-121 °, 84%.

oh c). · J-Nv.

° ^ KHj »j Vbu3 2 t-Bu

A suspension of 4.78 g (15 mmol) of cuproiodide in 50 ml of ether was cooled to 0 ° and treated under nitrogen with 26 ml of 1.9 M methyllithium solution (50 mmol). The brown solution was stirred at 0 ° for 10 minutes, then cooled to -60 ° and treated dropwise with 2.43 g (5.0 mmol) of trans-1- (tert-butyldimethylsilyl) -3-formyl-4-tritylthio. -2-acetidinone in a mixture of 10 ml of tetrahydrofuran and 40 ml of ether. Stirring was continued for 3 h. The solution was warmed to -40 ° and carefully treated with 1 M ammonium chloride solution. The mixture was filtered through Celite, the organic phase was washed with 3x5 ml of 1M ammonium chloride solution and then with brine and dried over sodium sulfate. Filtration and evaporation gave the alcohol, i.e. isomer B, which crystallized from pentane to give 1.6 g (65%), m.p. 160 ° -161 °, IR (CHCl 3) ν max: 1730 cm -1, 1 H MR (CDCl 3): δ 7.32 (15H, m), 4.05 (1H, s), 3.4 (1H, d, J = 3 Hz), 3.25-3.55 (1H, m), 1.6 (1H, s), 0.9 (12H, s) and 0.1 ppm (6H, s).

Note. a) tetrahydrofuran and ether were distilled off from LAH b) methyllithium was titrated with 1N hydrochloric acid c) copper (I) iodide was purified by continuous extraction with anhydrous tetrahydrofuran in a Soxhlet extractor for 18 h and then dried under vacuum in a desiccator for 18 h.

30 69845

OH

0 | 1 / SC0 1SC0

^ J CH, | J

> “N ^^ 2 ^ \ ς, · ^ o ^ Si 2 O Slx tBu tSu

0.1 ml (0.1 mmol) of methylmagnesium iodide was added dropwise to a cooled (0 °) and stirred solution of 25 mg (0.05 mmol) of trans-1- (tert-butyldimethylsilyl) -3-formyl-4- tritylthio-2-azetidinone in 2 mL of THF. The solution was stirred at 1.5 ° C, poured into anonium chloride solution, acidified with 1N hydrochloric acid solution and extracted with ether. Drying and concentration of the organic extracts gave an oil consisting of a starting material and a small amount of a mixture of two trans-title compounds with predominantly isomer B.

Example VI

(1'R, 3S, 4R and 11S, 3R, 4S) -11- (tert-butyldimethylsilyl) -3- (11-acetoxy-11-ethyl) -4-tritylthio-2-azetidinone (Isomer B )

OH

,! STr! . STr / '' --- ^ ....... '1 --- 1 Aco0

i / - ~ 7 -— r> X

N / pyrxdxxnx t \ Si 4- 0 x Si 4- \

\ X

A solution of 13.85 g (27.5 mmol) of (1'R, 3S, 4S and 1 ', S, 3R, 4S) -1- (tert-butyldimethylsilyl) -3- (11-hydroxy-11 -ethyl) -4-tritylthio-2-azetidinone in 75 ml of pyridine and 50 ml of acetic anhydride (prepared at 0 °), stirred at room temperature for 40 h. The reagents were removed by evaporation (the last residues were azeotroped three times with toluene). and an almost white solid remained. The crude derivative was crystallized from ether-petroleum ether to give the pure title compound (97.5%). 1 HMR (CDCl 3) {>: 7.64-7.03 (15H, m, H-aromatics), 4.60 (1H, m, J = 6, H-11), 3.92 (1H, d , 31 69845 J = 2, H-4), 3.55 (1H, dd, J = 2, J = 6, H-3), 1.79 (3H, s.

CH 3 CO), 0.98 (3H, d, J = 6, CH 3>, 0.88 (9H, s, t-butyl), 0.12 (6H, s, CH 3), IR (CHCl 3) max: 1775 , 1740 cm -1 (C = O).

Example VII

(1 ', R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-acetoxy-11-ethyl) -4-trityl-2-azetidinone (Isomer B) OAc OAc _ ^ Τγ NaN, _s STr

I --v. j I

-: N HMPT ^

0 ^ tBDMS

5.77 g (10.57 mmol) of pure derivative (1 ', R, 3S, 4R and 1'S, 3R, 4S) -1- (tert-butyldimethylsilyl) -3- (1'-acetoxy-1'- ethyl) -4-tritylthio-2-azetidinone in warm 60 ml HMPT and 10 no water. The solution was cooled to room temperature and 1.2 g of NaN 3 was added. After stirring for 45 minutes (the progress of the reaction was monitored by TLC) and 800 of any cold water was slowly poured. The mixture was stirred for 20 minutes. The crystalline material was isolated and washed with water. It was redissolved in its gels, washed twice with water and brine and dried over magnesium sulfate. Evaporation of the solvent gave a foam which was crystallized from ether-petroleum ether (4.90 g, 96.5%, mp 143 ° -144.5 °), IR (CH 2 Cl 2): 3395 (NH), 1772, 1738 cm -1 (C = 0), 1 H NMR (CDCl 3) δ: 7.9-6.8 (15H, m, H-aromatics), 5.12 (1H, dqtn center, J = 6.5, 7.5, H- 1 '), 4.33 (1H, d, J = 2.8, H-4), 4.20 (1H, broad, NH), 3.17 (1H, dd, J χ = 7.5, J3_4 = 2.8, 3-IgH = X 1 H (3)> 2.1 OH, s, CH 3 CO), 1.35 (3H, d, J = 6.5, CH 3).

Example VIII

(11, R, 3S, 4R and 1S, 3R, 4S) -3- (11-acetoxy-11-ethyl) -1- (p-nitrobenzyl-2 "-hydroxy-2" -acetate) -4-tritylthio- 2-acetidinone OAc OAc CHO \, STr .STr I r 1

- (C02PNB I

N TEA / 0 H co2pnb L.

69845 "Isomer B"

A solution of 1.82 g (30 mmol) of hydrated p-nitrobenzylglyoxylate (triturated in ether) in benzene was heated under reflux for 2 h through a Dean-Stark condenser filled with 3 Å molecular sieves. 10.88 g (25.2 mmol) of azetdinone- (1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-acetoxy-1'-ethyl) -4-tritylthio-2-azetidinone were added. and the mixture was heated under reflux for an additional 1 h. The solution was cooled to room temperature and 0.35 mL (2.5 mmol) of triethylamine was added. The mixture was then stirred for 2 h and the progress of the reaction was monitored by TLC. Evaporation of the solvent gave a white foam in quantitative yield (100%, epimer mixture). r * Alternatively, the solution may be washed with acid or base. IR (CH 2 Cl 2) νmax: 3520 (OH), 1775, 1745 cm -1 (C = O), 1 H NMR (CDCl 3): 8.2, 8.18 (2H, 2d, J = 8, Ho aromatic), 7.80-6.90 (17H, m, H-aromatic), 5.28, 5.17 (2H, 24, CH 2 -PNB), 4.89 (0.67H, d, J = 7.2, CHO), 4.80 (center of m, H-1 '), 4.38 (0.33H, 2d, J = 8.8, CHO), 4.22 (D, 33H, d , J 3 = 2.5, H-4), 4.09 (0.67H, d, J 4_3 = 2.1, H-4), 3.65 (D, 67H, dd, J χ = 5.8 J3_4 = 2.1, H-3), 3.47 (0.33H, dd, = 5.5, J3_4 = 2.5, H-3), 3.33 (0.33H, d, J = 8 , 8, OH), 3.23 (0.67H, d, J = 7.5, OH), 1.88, 1.86 (3H, 2s, CH 3 CD), 1.10, 1.06 (3H, 2d, J = 5.8, 6.3, CH 2.

Example IX

(1 ', R, 3S, 4R and 11S, 3R, 4S) -3- (1'-acetoxy-11-ethyl-1- (p-nitrobenzyl-2 "-chloro-2" -acetate) -4- tritylthio-2-acetidinone ° Ac OAc ^ STr STr —r socl2 i — Γ

N - ^ 0H pyridine ^ o N

O! υ! CO2PNB co2pnb "Isomer B" THF-Luo (distilled from LAH) containing (1 ', R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-acetoxy-1'-ethyl) - 1- (p-Nitrobenzyl-2 "-hydroxy-2" -acetate) -4-tritylthio-2-azetidinone (10.88 g of NH) was treated at -15 ° (ice-methanol bath) under nitrogen. 6 9 8 4 5 with 2.19 g (2.24 mL, 27.7 mmol) and 3.3 g (2.02 mL, 27.7 mmol) of thionyl chloride. The mixture was stirred for 20 minutes at -15 °. The salt was isolated by filtration and washed with benzene. Evaporation of the solvent (THF + benzene) gave a residue which was dissolved in warm benzene and treated with activated carbon. The suspension was filtered through a pad of Celite. The solvent was evaporated to give a foam. IR (CH 2 Cl 2) ν max: δ 80, 1740 cm -1 (C = O), 1 HMR (CDCl 3), δ: 8.17, 8.21 (2H, 2d, J = 8, Ho aromatic), 7, 76-6.88 (17H, m, H-aromatic), 5.31, 5.16, 5.12, 4.73 (3H, 4s, CH2-PNB, CHCl), 5.12-4.55 ( 1H, m, H1 '), 4.35-4.25 (1H, m, H-4), 3.80-3.45 (1H, m, H-3), 1.90 (3H, 3, CH 2 O), 1.12, 1.07 (3H, J = 6.5, CH 3).

Example X

(1'R, 3S, 4R and 11S, 3R, 4S) -3- (11-acetoxy-11-ethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio -2-azetidinone (isomer B) QAc OAc '"' W STr _? Al- ^) str 2,6-lutidine ^ --- ζ '^ V1 CO-PNB 1

2 0.02 PNB

A solution of impure (1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-acetoxy-1'-ethyl) -1- (p-nitrobenzyl-2 '-chloro-2'- --acetate) -4-tritylthio-2-azetidinone in 100 ml of dioxane (freshly distilled over LAH), treated with 2.97 g (3.23 ml,

27.72 mmol) with 2,6-lutidine and 9.91 g (37.8 mmol) of triphenylphosphine. The mixture was heated under reflux (oil bath, 130 °) for 18 h. The solvent was evaporated and the residue was redissolved in methylene chloride. The resulting solution was washed successively with dilute hydrochloric acid, water, dilute aqueous sodium bicarbonate solution, water and brine. Drying and evaporation of the solvent gave the title compound which was triturated with ether and isolated by filtration (14.6 g, 65.9%), IR

(CH_C1 ") \). : 1750 (C = O), and 1620, 1610 cm -1 (phosphorane).

22 max 69845 34

Example XI

(1 ', R, 3S, 4R and 11S, 3R, 4S) -3- (11-hydroxy-11-ethyl-1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio- 2-azetidinone (isomer B)

OAc OH

_ / STr NaOH N j - (/ STr i-Vp03 o '% -? I’--

6 C02PNB to2PNB

A solution of 4.43 g (5.00 mmol) of (1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-acetoxy-1'-ethyl) -1- (p-nitrobenzyl -2 "-triphenylphosphoranylidene-2" -acetate) -4-tritylthio-2-azetidinone in 10 ml of methanol and 60 ml of THF were treated at room temperature with 200 mg of 1% aqueous sodium hydroxide solution ( 1 equivalent in 20 ml of water), The reaction was monitored by TLC. The mixture was diluted with ether-ethyl acetate and washed with hydrochloric acid, water, aqueous sodium bicarbonate solution, water and brine. Evaporation of the solvent gave a residue which was crystallized from benzene ether to give 3.7 g (87.7%), m.p. 169.5 ° -170.5 °. IR (CH 2 Cl 2) δ: 1745 (C = O) and 1620 cm -1 1 2 max s (phosphorane).

Heating the mixture increased the reaction rate.

Example XII

(1'R, 3S, 4R and 1'S, 3R, 4S) -Silver-3- (11-hydroxy-11-ethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2- azetidinone 4-thiolate (isomer B)

oh 0H

/ AgNO3 v J J3Ag -r-STr C5H N = * ^ P * 3 O · "·",

O ν 'V

C02PNB C02PNB

II

35 6 9 8 4 5

A solution of + Y containing 1 g (1/19 mmol) of (1'R, 3S, 4R and 1'S, 3R, 4S) -3- (1'-hydroxy-1'-ethyl) -1- (p-nitrobenzyl- 2 "-triphenylphosphorylidene-2" -acetate) -4-tritylthio-2-azetidinone in 10 mL of methanol, treated with 124 μL (121.3 mg, 1.53 mmol) of pyridine and at 10 ° with 15 mL : 11a (2.25 mmol) of a 0.15 M methanolic solution of silver nitrate (or until silver mercaptide no longer precipitates). The mixture was stirred for 1 h and evaporated on a rotary evaporator (no bath) to a concentration of about 10%. The solvent was removed by filtration. The filter cake was washed once with methanol and three times with ether, sucked under high vacuum to give 954 mg (100%). IR

(nujol), 3500-3400 (O-H), 1752 (C = O), 1595 (phosphorane) mdK s and 1525 cm-1 (NO2> · +) The crystalline material was first dissolved in CH2Cl2. Preparation of final products Example 1 (1'R, 5R, 6S and 1'S, 5S, 6R) -6- (1'-hydroxyethyl) -2- (2-aminoethoxymethyl) -penem-3-carboxylic acid

OH

l ς j_s> ch2och2ch2nh2 co2h (1'R, 3S, 4R and 1'S, 3R, 4S) -4- (2-azidoethoxy) -acetylthio-3-11-hydroxyethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranyl -dene-2 "-acetate) -2-azetidinone OH' \ _ ^ SAg" '' r - f SSiMe-

/ | Me3SiCl / Et3N I J

0 <> · N- i P03 im?! HF ..... q ": '~‘ N ^ P03

_ CO „FNB

C02PNB OH 2

1} CV ^ ο ^ "Ν3 / ΡγΓ aS

--0- "j“ T Y vo ^ n3 2) TFA o Ν ^ γ ° ^ 3 co2pnb 36 69845

To a stirred solution of 820 mg (1.16 mmol) of (1'R, 3S, 4R and 1'S, 3R, 4S) -silver-3- (1'-hydroxyethyl) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone-4-thiolate in 20 ml of THF, 0.1648 ml (4.66 ml) were added successively at -15 ° (methanol-ice bath) under nitrogen. mmol, 4.02 equivalents) of triethylamine, 0.589 mL (4.64 mmol, 4.00 equivalents) of chlorotrimethylsilane and 81.2 mg (1.12 mmol) of imidazole. The mixture was stirred for 18 h (overnight) at room temperature and then cooled to -10 ° to 15 °. 0.220 mL (2.72 mmol) of pyridine was added followed by a solution of 372 mg (2.27 mmol, 1.96 eq) of 2-azidoethoxyacetyl chloride in 20 mL of CH 2 Cl 2. Stirred for 1 h at room temperature. After the precipitate was isolated by filtration, the filtrate diluted with EtOAc was washed successively with 1N hydrochloric acid, brine, saturated sodium bicarbonate solution and brine, dried over sodium sulfate, evaporated to give 748 mg of an oil.

This was dissolved in moist CH 2 Cl 2 (20 mL + 3 drops of water) and treated with 2 drops of trifluoroacetic acid for 30 minutes at room temperature. The mixture was washed with saturated sodium bicarbonate solution and then brine, dried over sodium sulfate, evaporated to give 695 mg of a crude oil.

This was purified by column chromatography (15 g SiO 2, eluent EtOAc-CH-ClO 1: 1) to give 538 mg (0.739 mmol, 63.7% yield) of the title compound as a yellowish oil, HMR (CDCl 3) δ: 1.22 (d , J = 6 Hz, CH 2 - (1), 5.6 (2d, H-4) and 7.3-8.4 ppm (aromatic H 5), IR (neat),: 3420 (OH), 2100 ( -N), 1750 (C = O) and 1690 cm -1 ITlClK 5 (thioester), R f = 0.20 (CH 2 Cl 2 -EtOAc 1: 1).

(1'R, 5R, 6S and 11S, 5S, 6R) -p-Nitrobenzyl 6- (11-hydroxyethyl) -2- (2-azidoethoxymethyl) penem-3-carboxylate (Isomer B)

OH N 0H

A, - / s O ^ toluene -T> S \ oANyp03 Δ 2 2 23

COjPNB C02PNB

II

69845

A solution of 49 mg (0.673 mmol) of (1'R, 3S, 4R and 1'S, 3R, 4S) -4- (2-azidoethoxy) -acetylthio-3- (1'-hydroxyethyl) -1- (p- nitrobenzyl 2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone in 80 ml of toluene, was gently heated under reflux for 3 h. Evaporation of the solvent in vacuo gave an oily residue which was purified by column chromatography (10 g SiO 2, eluent 5-10 CH 2 Cl 2). (CH 2 Cl 2) gave 202 mg (0.449 mmol, 66.8% yield) of the title compound as pale yellow crystals, HMR (CDCl 3): 1.35 (3H, d, J = 6.5 Hz). , CH 3-1 '), 2.18 (1H, broad, OH), 3.2-3.9 (5H, m, -CH 2 - and H-6), 3.9-4.5 (1H, m , H-1 '), 4.45-4.72-4.75-5.02 (2H, AB-type, CH2 ~ 2), 5.02-5.25-5.35-5.57 ( 2H, AB-type, -CH 2 Ar), 5.62 (1H, d, J = 1 Hz, H-5) and 7.42-7.65-8.13-8.28 ppm (4H, A2'B2 1, aromatic Hs), IR (nujol) ν max: 3460 (-OH), 2210 (-N 2), 1765 (β-lactam) and 1680 cm -1 (ester). An analytical sample was obtained by continuing crystallization, m.p. 107 ° -108 ° (CH 2 Cl 2 -ether), UV (EtOH) λ max: 264 (E = 1200 °) 3a 323 m; u (E = 9200), R f = 962 (CH 2 Cl 2 -EtOAc 1: 1), analysis C 18 H 19 N 5 O 7 S , calculated: C 48.10, H 4.26 N 15.88 S 7.13, found: C 47.81 H 4.18 N 15.00 S 7.16.

(1'R, 5R, 6S and 1 * S, 5S, 6R) -6- (1'-hydroxyethyl) -2- (2-aminoethoxymethyl) penem-3-carboxylic acid

^ OH ^ OH

JZfy ~ ™ 2 ° “2 ° * 2« 3 --- o JrO ^ CH2 ° CH2CH / H2

C02? MB C02H

A solution of 180 mg (0.400 mmol) of (1'R, 5R, 6S and 1'S, 5S, 6R) -p-nitrobenzyl 6- (1'-hydroxyethyl) -2- (2-azidoethoxypenem-3-carboxylate) was stirred In 18 ml of THF, 19 ml of ether, 18 ml of water and 180 mg of 10% palladium on carbon Hydrated (H, 3.85 kp / cm) for 2.5 h at room temperature Catalyst 38 69845 removed by filtration, the aqueous filtrate was washed with EtOAc, lyophilized to give 84.4 mg (0.293 mmol, crude yield 73.2%) of the title compound as an impure, yellowish powder.

UV (H 2 O) λ max: 305.5 (i = 4800.) and 255 mp (£ = 3800). This powder was purified by HPLC (Waters C1Q Micro Bondapack

Reverse Phase 30 cm x 10 mm, eluent 1% CH 2 Cl 2 in water) to give 44.7 mg (0.155 mmol, 38.8% yield) of the title compound as a white powder. 1 HMR (D 2 O) δ: 1.34 (3H, d, J = 6.4 Hz, CH 2 Cl 2), 3.26 (2H, m, -CH 2 N), 3.82 (2H, m, -OCH 2 CH 2) -), 3.94 (1H, dd, J6_1 = 6.2

Hz, J 1 = 1.4 Hz, H-6), 4.2-4.4 (1H, m, H-1 '), 4.52-4.70-4.84-b — b 5.02 ( 2H, AB-type, CH 2 22) and 5.71 ppm (1H, d, J = 1.3 Hz, H-5), IR (KBr tablet) J max: 3420 (OH), 3000-2600 ( broad, CO 2 H), 1765 (/ Wactam) and 1575 cm -1 (-CO 2 H), UV (H 2 O) λ max: 306 (5300) and 258 myi (i. = 3600).

Example 2 2- (2-Aminoethoxymethyl) -penem-3-carboxylic acid (via mercaptide intermediate) J-2 2 2 2 co2h

Ethyl 2-chloroethoxyacetate F tO Π VTEAB __ ^ 150-160 ° C o

II

A mixture of 24.5 g (0.200 moles) of ethyl chloroacetate, 8.80 g (0.300 moles) of ethylene oxide and 0.40 g (1.9 mmol, dried in vacuo) of tetraethylammonium bromide was heated in a reactor vessel 39,69845 for 6 h at 150 ° At -160 °. After cooling, the reaction mixture was distilled in vacuo to give 6.66 g (54.4 mmol, 27.2%) of ethyl chloroacetate, b.p. 22 ° -24 ° (0.5 mm Hg) and 8.39 (50.4 mmol, 25.2%) ethyl 2-chloroethoxyacetate as a colorless oil, b.p. 49 ° -53 ° (0.1 mm Hg), 1 H NMR (CDCl 3): 1.28 (3H, t, J = 7 Hz, -CH 3), 3.5-4.0 (4H, m, N -CH 2 CH 2 -Cl), 4.15 (2H, s, -COCH 2 O-), 4.25 ppm (2H, q, J = 7 Hz, -OCH (CH-) IR (neat) 0: 1740 cm -1 1 δ 3 max (C_0-ester). Method: D Klamann et al., Justus Liebig Ann., 710, 59 (1967) (reported: yield 42%, b.p. 55.5 ° / 0.35 mm Hg). Ethyl 2-azidoethoxyacetate r ·! .. »t EtO N_ 1 O DMF o o

A mixture of 7.71 g (36.3 mmol) of ethyl 2-chloroethoxyacetate and 3.31 g (50.9 mmol) of sodium azide in 100 ml of DMF was heated at 80 ° -90 ° C for 3.5 h. At which time TLC (hexane-ether 1: 1) indicated the reaction was complete. The cooled mixture was poured into 1 liter of water and extracted with 3 x 250 ml of ether. The extracts were washed twice with water and once with brine, dried over magnesium sulfate, evaporated to give 7.16 g (41.4 mmol, 89.4%) of ethyl 2-azidoethoxyacetate as a yellowish oil. 1 H NMR (CDCl 3%: 1.30 (3H, t, J = 7 Hz, -OCH 2 CH 3), 3.3-4.0 (4H, m, -OCH 2 CH 2 N 3), 4.13 (2H, s, -COCH 2 O-) , 4.23 ppm (2H, q, J = 7 Hz, -OCH_CH_), IR (neat),: 2100

^ j ITI cl S

(N3) and 1750 cm (C = O-ester). This material was used in the next step without further purification.

2-atsidoetoksietikkahappo

EtOAc, NaOH HO2N

HgO-MeOti * 3

o O

To a solution of 6.56 g (37.9 mmol) of ethyl 2-azido-40 6 9 8 4 5 ethoxyacetate in 80 ml of methanol was added 80 ml of 1N aqueous sodium hydroxide solution, and the mixture was stirred at room temperature overnight (17 h). . After removal of insoluble material, the methanol was evaporated in vacuo and the residue was saturated with sodium chloride and washed with 3 x 30 ml of ether. The aqueous layer was acidified with 30 mL of 3N HCl and extracted with 4 x 40 mL of ether.

The ether extracts were washed with brine, dried over magnesium sulfate, evaporated to give 4.25 g (29.3 mmol,

77.3%) 2-azidoethoxyacetic acid as a colorless oil. ^ HMR

(CDCl 3) δ: 3.3-4.0 (4H, m, -ΟΟΗ, β 2), 4.22 (2H, s, -COCH 2 O-), 9.52 ppm (1H, s, -CO (H, converted to DO), IR (neat), 2. zm ci K. s 2600-3300 (broad, -CO 2 H) 2100 (azide) and 1740 cm (C = O-CO 2 H).

This material was used in the next step without further purification.

2-Azidoethoxyacetyl chloride “γΛοΑ / ^ soci2 Ci

o O

A solution of 2.09 g (14.4 mmol) of 2-azidoethoxyacetic acid in 5 ml of thionyl chloride was stirred for 4 h at room temperature. Excess thionyl chloride was removed in vacuo from a water suction pump and the residue dissolved in 10 ml of benzene (dried on a molecular sieve) was evaporated in vacuo. The oil thus obtained was dried in vacuo (water suction pump) over NaOH to give 2.23 g (13.6 mmol, 94.4%) of 2-azidoethoxyacetyl chloride as a colorless oil. 1 HMR (CDCl 3) δ: 3.43 (2H, broad, t, J = 5 Hz, -CH 2 O-), 3.78 (2H, broad, t, J = 5 Hz, -CH 2 N 3) and 4, 50 ppm (2H, s, -COCH 2 O-), IR (neat) (max) 2100 (azide) and 1800 cm -1 (-COCl). This material was used in the next step without further purification.

Il 41 69845 4- (2'-Azidoethoxyacetylthio) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2w-acetate) -azetidinone _ / SA9 Pyr / CH Cl aY / si. I -2-u n s

C02PNB CO 2 PNB

To a stirred solution of 7.96 g (12.0 mmol) of silver 1- (p-nitrobenzyl) -2'-triphenylphosphoranylidene-2'-acetate) -2-azetidinone-4-thiolate in 100 mL of CH2Cl2 A solution of 2.23 g (13.6 mmol) of 2-azidoethoxyacetyl chloride in 20 ml of 1.94 ml (24.0 mmol) of pyridine was added at 0 ° -5 ° and under nitrogen. CH 2 Cl 2 and the mixture were stirred for 2 h at room temperature. After the precipitate was removed by filtration, the filtrate was evaporated, the residual oil was purified by column chromatography (160 g SiO 2 eluent EtOAc-CH 2 Cl 2 1: 1) to give 4.216 g (6.17 mmol, 51.4%) of the title phosphorane as a yellowish foam. This was used in the next step. An analytical sample was obtained by crystallization from CH 2 Cl 2 -ether (1: 9), m.p. 128 ° -129 ° (decomposes), IR (nujol) δ max: 2090 (-N 2), 1755 (β-lactam) and 1690 cm -1 (thioester) Analysis C 34 H 30 N 5 O 7 PS Calculated: C 59.74 H 4 , 42 N 10.26 S 4.69, found: C 59.33 H 4.49 N 9.69 S 5.19 TLC (EtOAc): Rf = 0.55.

p-Nitrobenzyl 2- (2-azidoethoxy) methylpenem-3-carboxylate _ / N.

3 s I-f v toluene -— \

ö ---- yb — CH OCH CH N

P * 3 4

I CO PNB

C02PNB 2 42 69845

A solution (cloudy) of 4.13 g (6.04 mmol) of the above phosphorane in 200 ml of toluene was heated under reflux for 1.5 h under nitrogen. After removal of the insoluble matter, the solvent was evaporated in vacuo, and the residual oil was purified by column chromatography (80 g SiO 2, eluent 5% ether in benzene) to give 2.44 g (6.02 mmol, 99.6%) of the title compound as a yellowish oil. This was used in the next step. Crystallization from CH 2 Cl 2 (1: 9) gave an analytical sample, m.p. 88 ° -89.5 °, 1 H NMR (CDCl 3) δ: 3.35 (2H, t, J = 5 Hz, -OCH 2 -), 3.47 (1H, dd, J = 16 Hz, J. = 2 Hz , C, -H), 3.67 (2H, t, J = gem trans 6 '5 Hz, -CH-N-,), 3.85 (1H, dd, J = 16 Hz, C -H), 4.73 (2H, ABq, J = 23 gem 6 16, 19, C2-CH2), 5.30 (2H, ABq, J = 13.5, 9, -OCH2Ar), 5.63 (1H, dd, J. = 2 Hz, J. = 3.5 Hz, C - H), 7.50-7.63-8.12-8.27 ppm (4H, A2'B2 ', aromatic Hs), IR ( nujoli) v) majcs: 2100 (-N2), 1785 (β-lactam) and 1695 cm-1 (ester), UV (EtOH) 263 ιημ (£ = 12,000), 320.5 myi (£ = 9600), TLC (benzene-ether 1: 1):

Rf = 0.60.

2- (2-Aminoethoxy) -methylpenem-3-carboxylic acid N-CH2CH2CH2N3- (CH2OCH2CH2NH2)

F M3 CO H

2 2

A solution of 1.62 g (4.00 mmol) of the above azido ester in 50 ml of dimethoxyethane was mixed with 50 ml of ether, 50 ml of water and 1.62 g of 10% palladium on carbon. carbon (Engelhard), and hydrogenated for 2.5 h at room temperature (H ~: 3.85 o Δ kp / cm). After the catalyst was removed by filtration of the aqueous layer, 69845 4 3 was washed with 2 x 50 mL of ether and then 1 x 50 mL of EtOAc. The aqueous layer was lyophilized to give 817 mg (3.34 mmol, 83.6%) of the title amino acid as a yellowish powder, UV (Η2 <3) λ max: 304 mY (£, = 5000). This material was purified by HPLC (Waters, C ? 3.88 (1H, dd, J = 16.8 Hz, J = 3.7.Hz, C 1 H), 4.52-4.70- gem cxs o 4.83-5.01 (2H , AB type, ¢, -01.0-) and 5.77 ppm (1H, dd, Jcls = 3.6

Hz-Jtrans (KBr tablet) max 1770 (β-lactam) and 1580 cm (-CO 2 H), UV (H 2 O) max: 304 μm (i = 5400), 256 μg (t = 3100).

Example 3 2- (2-Aminoethylthiomethyl) -penem-3-carboxylic acid (via the mercaptide intermediate) -rsv <^ L ^ ch * scw "'

COOH

2-atsidoetyylimetaanisulfonaatti

BrCH2CH2OH -► N3CH2CH2OMs

A solution of 7.5 g (60.0 mmol) of bromoethanol and 5.0 g (76.9 mmol) of sodium azide in 30 mL of HMPT was heated at 115 ° for 2.5 h. The reaction mixture was cooled to 23 ° and diluted with 100 mL of CH 2 Cl 2. The solid was removed by filtration, the CH 2 Cl 2 was evaporated on a rotary evaporator to leave a yellow liquid which was cooled to 0 ° and treated sequentially with 5.57 mL (72.0 mmol) of mesyl chloride and 10.0 mL (72.0 mmol) of triethylamine. The reaction mixture was stirred for 1 h at 0 ° and then for 6 h at 23 ° was poured into 300 ml of water. The aqueous solution was extracted with 1 x 200 mL, 4 x 100 mL of ether, the ether extracts were combined, washed with 1 N HCl, water, saturated 4 "6984 5 sodium bicarbonate solution and water, dried over anhydrous magnesium sulfate and evaporated on a rotary evaporator. under high vacuum, bp 95 ° -100 ° / 0.3 torr, 5.8 g, 58.5%, IR (neat) δ,: 2005, _ 1 ·, max (s, N 3), 1345, (s, SO 2 -O), 1175 (m, SC 2 -O) cm -1, HMR (CDCl 3) δ: 3.03 (s, 3H, OCH 3), 3.43-3.76 (m, 2H, H-2) and 4.2-4.46 ppm (m, 2 H, H1).

21-Azidoethylthioglycolic acid! Δ

N3CH2CH2OMs + NaSCH2COONa-β ^ N- ^ CH 2 H ^ CH 2 OOH

2. B

3.14 g (34.1 mmol) of thioglycolic acid were treated with 68 ml (68.0 mmol) of a 1N NaOH solution, the resulting solution was stirred for 0.5 h at 23 ° and treated with a solution of 5.3 g (32 g). , 1 mmol) of 2'-azidoethyl methanesulfonate in 15 ml of dimethoxyethane. The reaction mixture was stirred at 45 ° for 22 h, cooled to 23 °, washed with 3 x 20 mL of CH 2 Cl 2, acidified with 6 N hydrochloric acid, and extracted with 7 x 40 mL of CH 2 Cl 2. The CH 2 Cl 2 extracts were combined, dried over anhydrous sodium sulfate and evaporated on a rotary evaporator to an oil which was distilled under high vacuum, b.p. 117 ° -122 ° / 0.27 torr, 4.2 g, 81.2%. IR (neat) ν max = 2100 (s, Nj), 1708 (s, C = O) cm -1, 1 HMR (CDCl 3) δ: 2.7-3.07 (m, 2H, H-1 ') , 3.35 (s, 2H, H1), 3.30-3.73 (m, 2H, H-2 ') and 11.81 ppm (s, 1H, COOH).

2'-azidoethylthioacetyl chloride (COCl) ^ p-, pj, nnj, cnpl N3CH3CH3SCH3COOH ---> iN3 2 2 2 ^^

To a solution of 3.33 g (20.7 mmol) of 2-azidoethylthioglycolic acid in 50 mL of CH 2 Cl 2 was added 3.9 mL of oxalyl chloride and 1 drop of DMF. The reaction mixture was stirred at 23 ° for 1.5 h, the solvent was removed on a rotary evaporator to give a yellow solid.

liquid. IR (neat) δ: 2100 (s, N 1), 1785 (broad, C = O). ^ HMR

ms k s J

(CDCl 3) δ: 2.6-3.0 (m, 2H, H-1 '), 3.37-3.73 (m, 2H, H-2') and 3.82 ppm (s, 2H, hl).

45 69845 4- (21-Azidoethylthioacetylthio) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone

^ SAg 5CCH.SCH CH N

-S ClCCH_SCH_CH_N, -2 2 2 3

I _2_2 2 „J-► I

0 ^ j> pph3 σ Nj: = pph3

COOPNB COOPNB

A solution of 15.7 mmol of silver 1- (p-nitrobenzyl 1'-triphenylphosphoranylidene-1'-acetate) -2-azetidinone-4-thiolate and 1.6 mL (19.8 mmol) of pyridine 200 in 1 mL of CH 2 Cl 2 was treated dropwise with a solution of 3.64 g (20.3 mmol) of 2'-azidoethylthioacetyl chloride in 50 mL of CH 2 Cl 2. The reaction mixture was stirred for 1.5 h at 23 °, filtered and the solid was washed with CH 2 Cl 2. The filtrate and washings were combined and washed with 0.1 N HCl solution, water, saturated sodium bicarbonate solution and water, dried over anhydrous sodium sulfate and evaporated on a rotary evaporator to give an orange syrup. Column chromatography (300 g silica gel G-60, eluent EtOAc in CH 2 Cl 2, 0-40%) gave 7.7 g, 70% of the crude compound after evaporation of the solvent. An analytical sample was obtained by recrystallization from CH 2 Cl 2-ether-petroleum ether, m.p. 150 DEG-151 DEG C., decomposes. Analysis for C34H3QN2O2S2P Calculated: C 58.36 H 4.32 N 10.01 S 9.17, found: C 58.64 H 4.36 N 10.03 S 9.25, IR (KBr) ^ maJcs: 2100 (s »N 3), 1750 (s, β-lactam C = 0), 1675 (s, C = O), 1655 (s, C = O), 1610 (s, aromatics) and 1440 cm -1 (s, P -Ph).

p-Nitrobenzyl 2-azidoethylthiomethylpenem-3-carboxylate

s? ch2sch2ch2n3 r — rV

Π - ^ - V CH 2 SCH 2 CH 2 N 3 ° Y-PPh 3 0 ΛοΟΡΝΒ

COOPNB

46 69845

A suspension of 4.5 g (6.43 mmol) of 4- (2'-azidoethylthioacetylthio) -1- (p-nitrobenzyl-2 "-triphenylphosphoranylidene acetate) -2-azetidinone in 375 ml of toluene was stirred. 2.25 h at 110 ° under nitrogen, the reaction mixture was cooled to 23 ° and the solvent was evaporated on a rotary evaporator to give an orange syrup. The crude material was purified on a silica gel column (90 g of silica gel G-60, eluent ether-petroleum ether ether 1: 1-3). The pure material was obtained as a yellow syrup, 2.2 g, 81% IR (neat) 0 ma] <s: 2100 (s, N 2), 1785 (s, β-lactam C = 0), 1705 cm -1 1 (s, C = 0 of PNB), 1 H NMR (CDCl 3) δ: 2.53-2.90 (m, 2H, H1 "), 3.30-3.67 (m, 3H, H 2 O) ", H-6 trans), 3.98 (ABq, J 1 = 14.8 Hz, 2H, H-11), 5.32 (ABq, Ja_b = 13.0 Hz, 2H, CH 2 ~ PhNO 2), 5 , 66 (dd, JH_5 R_6 cis = 3.6 Hz, JH_5 / H_6 trans = Hz '1_H'H-5>' 7'58 <d 'JHo-Hm = 8.8 Hz, 2H, Ho, PNB) and 8.19 ppm (d, J = 8.8 Hz, 2H, Hm, PNB).

ηΓΠ “ηΟ 2-aminoethylthiomethylpenem-3-carboxylic acid I V-CH SCH ca N -> I Γ i-CH SCII CH tlH * J-νΎ 2223 coopnb co6 ^

To a solution of 45 mg (0.11 mmol) of p-nitrobenzyl 2-azidoethylthiomethylpenem-3-carboxylate in 5 mL of dimethoxyethane was added 5 mL of ether, 5 mL of water, and 45 mg (0.11 mmol) of 10% of palladium carbon. The reaction mixture was hydrogenated at 23 ° 2 under a hydrogen pressure of 3.15 kp / cm for 3.0 h and filtered through a pad of Celite. The layer was washed with water, the filtrate and washings were combined and diluted with ether. The aqueous phase was isolated, washed with ether and lyophilized. The crude product (20 mg) was purified by HPLC, 5 mg, 18%. IR (KBr) ν max: 1765 (C = 0.1600 cm -1 (broad, COO_), 1 HMR (D 2 O) δ: 2.70-3.00 (m, 2H, H1 "), 3.15-3 .45 (m, 2H, H-2 "), 3.49 (dd, Jgem = 16.8 Hz, J6 ^ trans = 1/7 Hz, H-6 trans), 3.85 (dd, J = 16 , 8 Hz, j, c. = 3/4 Hz, H-6 cis), 4.05 (ABq. Da_b = 14.6 Hz, 2H, H1) and 5.74 ppm (dd, cis = 3.4 Hz,

II

69845 J5-6 trans = Χ'7 Hz '1H' H "5) 'UV λ max: 307 (& = 4330)' 250 (i, = 3282).

Example 4 2- (2-Aminoethylsulfonylmethyl) -penem-3-carboxylic acid Ich ^ Ich ^ ch ^ nh ^ 2 2 2 2

COOH

p-Nitrobenzyl 2- (2-azidoethylsulfonylmethyl) penem-3-carboxylate

Γ \ —CH SCH CH N -MCPBA- CH JicH CH N

2 2 2 3 CH 2 = 2 23

COOPNB COOPNB

A solution of 0.36 g (0.85 mmol) of p-nitrobenzyl 2-azidoethylthiomethylpenem-3-carboxylate in 30 ml of CH 2 Cl 2 was cooled to -20 ° under nitrogen and treated dropwise with a solution of 0.147 g for 2 h. g (0.85 mmol) of m-chloroperbenzoic acid in 90 ml of CH 2 Cl 2. The reaction mixture was stirred at -20 ° for 0.5 h, warmed to room temperature and washed with saturated sodium bicarbonate solution and water. The organic solution was dried over anhydrous sodium sulfate and evaporated in vacuo to an orange residue which was chromatographed on silica gel (9 g silica gel G-60, eluent 25% EtOAc in CH 2 Cl 2, fraction volume 7 mL). Evaporation of the appropriate fractions gave 0.27 g of a white solid, m.p. 128 ° -131 °, 72.6%. Recrystallization from acetone-ether-petroleum ether gave an analytical sample, m.p. 142 °, decomp. Analysis Calculated: C 43.93 H 3.46 N 16.01 S 14.66, found: C 43.79 H 3.44 N 16.02 S 14.63, IR (KBr) + max: 2110 (N3 ), 1785 (β-lactam C = 0), 1690 (PNB ester C = 0), 1600, 1560 (C = O), 1520, 48 69845 1355 cm -1 (NO 2), uv-max 3: 265 (t (C = 12884) (33 = 8764), 1 H NMR (CDCl 3) δ: 2.95 (2H, m, CH 2 H 2), 3.58 (dd, JR_6_H_5 trans = 2.0 Hz, J = 16.6 Hz, H-6 trans), 4.33 (center of ABq, J = gem a, b 13.4 Hz, H-11), 4.32 (center of ABq, J, = 13.2 Hz , H-11) 5.33 3. f Ö (center of ABQ, = 13.7 Hz, 2H, CH of the PNB ester), 5.75 (dd, JH-5-H-6 cis = 3 '^ HZ / JH-5-H-6 trans = 2.1 Hz '1H' H_5) '/, 6 ° (d, J' '= 8.8 Hz, 2H, Ho of PNB ester) and 8.22 (d , JTT „- 8.8 rio-Hm Hm-Ho

Hz, 2H, Hm of the PNB ester).

2- (2-Aminoethylsulfonylmethyl) -penem-3-carboxylic acid __s ° 10% Pd / c, -γ sx ?, I> CH0SCH0CH0N4 - ^ T - r-7r „n -N ~ NH9 .-, / 7 2 2 2 3 DMe, Eto0, Ho0> —N // 2 2 2 2

^ \ X0PNB

To a solution of 57 mg (0.13 mmol) of p-nitrobenzyl 2- (2-ethylsulfonylmethyl) penem-3-carboxylate in 20 ml of dimethoxyethane were successively added 10 ml of E 2, 10 ml of water and 57 mg of % palladium on carbon. The reaction mixture was hydrated for 1.25 h at 3.85 kp / cm and filtered through a pad of Celite. The filtrate was diluted with Et 2 O, the organic phase was isolated, the aqueous solution was washed twice with Et 2 O and lyophilized. The crude orange powder (30 mg) was purified by HPLC. Lyophilization of the appropriate fractions gave the title compound as a white powder, 10.4 mg 29%, υνΛ; 313 (<= 4877), IR (KBr) v max: 1720 </; β-lactam C = O) and 1590 (carboxylate), 1 HMR (D 2 O) δ: 3.0-3.7 (5H, H-6 trans, CH 2 CH 2 NH 3 +), 3.90 (dd, JH_6_H_5 cis = 3.6 Hz, Jgem = 16.9 Hz, 1H, H-6 cis), 5.45 (center of ABq, J, = 13.6 Hz, H-1 '), 4.50 (center of ABq, J , = 13.6 Hz, H-1 ') and 5.8 (m, 1H, H-5).

cl / O

II

69845 49

Example 5 (1'R, RAS and 1'S, 5S, 6R) -6- (1'-hydroxyethyl) -2- (2-aminoethoxymethyl) -penem-3-carboxylic acid

OH

s \

CO 2 H

(1'R, 3S, 4R and 1'S, 3R, 4S) -4- (2-azidoethoxyacetylthio) -3-1'-hydroxyethyl) -1- (p-trimethylsilylethyl-2 "-triphenylphosphoranylidene-2" - acetate) -2-azetidinone OH 90 J—, P0, ^ 0> 'Η' ύ- · ° Y 3 0 I: «e

1.54 ml (11.8 mmol) of trimethylsilyl chloride were added to a stirred suspension of 2.48 g (3.34 mmol) of silver-3- (1'-hydroxyethyl) -1- (β-trimethylsilylethyl). 2 "-triphenylphosphoranylidene-2" -acetate) -2-azetidinone thiolate, 136 mg (2.0 mmol) of imidazole and 1.64 mL (11.8 mmol) of triethylamine in 60 mL of THF ° C. The mixture was stirred for 18 h at 23 °. 60 ml of methylene chloride were added, the mixture was cooled to -15 °, 1.32 ml (16.4 mmol) of pyridine and 1.43 g (8.70 mmol) of γ-azidoethoxy chloride were added and the mixture was stirred for 0.5 h at -15 °. C. 60 ml of ether, 60 ml of ethyl acetate, i.e. 69845, and 20 ml of 1M hydrochloric acid were added. The precipitate was removed by filtration and the organic phase was washed with 100 ml of 0.1 M hydrochloric acid, 100 ml of 1% sodium bicarbonate solution and saturated sodium chloride solution. Evaporation of the dried solution gave the crude title product as an oil, yield 85%. IR max: 1755 and 1695 cm-1.

(1'R, 5R, 6S and 1'S, 5S, 6R) - [3-Trimethylsilylethyl 2- [3-azidoethoxymethyl-6- (11-hydroxyethyl) -penem-3-carboxylate

OH

i_ ^ SCOC,! 2OCH2CH2 »:) A -s, -i ΡΦ -— Π / -ch2oc« 2o «2n3», Sl— pr-) y \ y * 3 ^ co2 / ^ / \ \

A solution of 1.3 g of the above phosphorane in 200 ml of toluene was heated under reflux for 3 h. Evaporation of the solvent on a rotary evaporator gave the crude title product. Chromatography on 40 g of silica gel eluting with increasing amounts of ether in hexane gave the crystalline title compound in 65% yield. IR 1 imgg: 1760 and 1700 cm -1, 1 HMR showed that the product was contaminated with the second isomer.

(1'R, 5R, 6S and 11S, 5S, 6R) -2- /% -Azidoethoxymethyl-6- (11-hydroxyethyl) -penem-3-carboxylic acid

OH

OH

A.-f-S \, 1

0 CO H

co2 / N / \ 2

A solution of 3 mL (1.5 mmol) of anhydrous tetrabutylammonium fluoride in THF was added to a solution of 155 mg (0.37 mmol) of the above ester in 2 mL of THF at 0 °. After 5 minutes at 0 °, 10 ml of water and 10 ml of ethyl acetate were added, the mixture was acidified to pH 3 with 1M hydrochloric acid and the phases were separated. The organic phase was extracted with 0.05 M sodium bicarbonate solution, the aqueous extracts were acidified to pH 3 with hydrochloric acid and extracted with ethyl acetate. The organic extracts were washed with saturated sodium chloride solution, dried, evaporated on a rotary evaporator, the residue triturated with ether to give 27 mg of the crude title compound as a solid, yield 28%. * R l? max: 3500, 1785, 1670 cm-1, 1 H NMR (CDCl 3), δz 1.30 (3H, d, J = 6.5, CH 3-1 '), 2.22 (1H, OH), 3.1- 3.9 (5H, m, CH 2 and H-6), 3.9-4.4 (1H, m, H-1 '), 5.60 (1H, d, J = 1, H-5).

(1'R, 5R, 6S and 11S, 5S, 6R) -6- (11-hydroxyethyl) -2- (2-aminoethoxymethyl) penem-3-carboxylic acid

oh QH

CO H CO H

2 * -

A solution of 150 mg of the above azido compound in 15 mL of THF, 15 mL of ether and 15 mL of water was hydrogenated.

In a Parr shaker with 150 mg of 10% palladium-carbon on an initial pressure of hydrogen 2 of 4.2 kp / cm. After 3 hours, the catalyst was removed by filtration through Celite, the aqueous phase was washed with ethyl acetate, lyophilized to give the crude title compound. Purification by HPLC (Waters, C 2 g Micro Bondapack Reverse Phase) gave 46.7 mg of pure title compound, which was the same as the previously prepared sample prepared by hydrogenation / hydrogenolysis of the corresponding azido-p-nitrobenzyl derivative.

52 69845

Example 6 6-Ethyl-2- (2-aminoethoxymethyl) -penem-4-carboxylic acid C2H5.

V CH OCH CH JH

J-N \ J /

0 XCO2H

^ OAc ^ OAc -'0Ac CSI, '"I f + Y ~ |

NaHCO 3 cT “NH + </ ~ m

Na 2 O 3

I II III

1-Butenyl acetate (a mixture of cis and trans isomers of about 1: 1) was prepared by P.Z. Bedoukian, J. Am. Chem. Soc., 66, 1325 (1944).

To 50 mL of cooled (-15 °) Compound I was added dropwise 10 mL (11 g, 78 mmol) of CSI. The mixture was allowed to gradually warm to 0 ° over 30 minutes. It was cooled to -20 ° and carefully poured into a mixture of 8 ml of water, 35 g of ice, 18.4 g of sodium bicarbonate and 6.4 g of Na 2 SO 3. The mixture was stirred vigorously for 30 minutes at 0 °, treated with 250 mL of petroleum ether and cooled to -40 °. The solvent was removed by decantation and the residue was treated in the same manner with 100 ml of petroleum ether. The combined petroleum ether extracts were washed with 30 ml of water and dried over sodium sulfate to recyclize compound I.

The aqueous phases were combined and extracted with 5 x 40 ml of ethyl acetate. The extract was dried over sodium sulfate, evaporated in vacuo to give 7.0 g (57%) of a mixture of 28% II and 72% III, b.p. 82 ° -85 ° (0.01 mm Hg), NMR δ (ppm, CDCl 3): 7.3 (1H, NH), 5.92 (0.72H, d, J = 4.4, II-H- 3), 3.3 (0.28H, d, J = 1.4 III-H-4), 3.3 (1H, m, H-3), 2.24 (3H, s), 2.72 (2H, two, Q, J = 7), 1.1 (3H, two t, J = 7), ^ c = o: 1775, 1755 cm Analysis: C ^ H ^ NO. ^, Calculated: C 53.49 H 7.05 N 8.91, found: C 53.12 H 6.93 N 8.85.

li 69845 53 .QCOCH ^ NaSCOCH COCl 3

__NH * J- NH

II + III IV

Sodium thioacetate was prepared by adding 0.8 mL (850 mg, 11.2 mmol) of 1 N sodium hydroxide solution in 11.2 mL of cooled (ice bath) under nitrogen. This was added to a cooled solution of 1.57 g (10 mmol) of compounds II and III in 5 ml of water under nitrogen. The mixture was stirred for 1 h at room temperature.

As the oil separated, 9 ml of acetone were added and stirring was continued for 1.5 h. The mixture was evaporated in vacuo to remove acetone and then extracted with methylene chloride. The extract was dried, evaporated in vacuo to give 1.65 g (95%) of a crude mixture of 85% trans-IV and 15% cis-IV, b.p. 105-110 ° (0.02 mm Hg), 7.1 (1H, NH), 5.53 (0.23H, d, J = 4.5, cis-H-4), 5.12 (0, 8H, d, J = 2.4, trans-H-4), 3.34 (1H, two t, J = 7), 2.48 (3H), 1.9 (1H, two q, J = 7) ), 1.15 (3H, two, t, J = 7) (c) Q: 1700, 1765 cm -1 Analysis for C7H11N 'calculated: c 48.53 H 6.40 N 8.07, found: C 48.18 H 6.47 N 7.77.

. SCOCH / ΊΛ A> Ac - (3 07N- \ Q / -CH2-CO2CH (OH) 2 | J "" NH o '^ ~ N \ choh

tv v I

C02PNB

A mixture of 1.25 g (7.2 mmol) of compound IV and 1.6 g (7.5 mmol) of p-nitrobenzylglyoxylate in 80 ml of benzene was heated under reflux for 20 h under a Dean-Stark water separator, then evaporated in vacuo to give 3.01 g of crude product. This was isolated by filtration through a small amount of silica gel to give 2.8 g (quantitative yield) of a pale yellow oil V containing a slight solvent, <f 7.9 (4H, m), 5.3 (4H, m) , 4.8 (1H, OH), 3.2 (1H, m), 3.37 and 3.33 (3H, two s), 1.8 (2H, m), 1.05 (3H, m) , l) c: 1765, 1700 cm This product was used in the next step without further purification.

^ ^ scoch3 soci? _ ^ Mv .- ^ coch3 NV Q N V'vHCl 1 ^ CHOH |

| CO PNB

CO PNB 2

V VI

To a cooled (ice bath) and stirred solution of 2.1 g (5.5 mmol) of compound V in 10 ml of dry benzene was added 3 ml of thionyl chloride, the mixture was kept at 5 ° for 2 h and evaporated in vacuo at room temperature. Excess thionyl chloride was removed by adding and evaporating benzene several times, the product was purified by filtration of the benzene solution through a small amount of silica gel to give, after evaporation in vacuo, 1.7 g (77%) of crude pale yellow oil VI, 7.9 (4H, m), 6.0 (1H, s), 5.3 (3H, m), 3.3 (1H, m), 2.7 and 2.3 (3H, two s), 1.75 (3H, m), 1.0 (3H, m), λ max = 0: 1700, 1775 cm The product was used without further purification in the next step.

/ V. / S-COCH

V Ml

I> J- N

"-CHCl 11 Utidine / T * 3

I CO PNB

CO PNB 2 2

VI

VII

II

55 6 9 8 4 5

A mixture of 1.7 g (4.2 mmol) of compound VI, 1.57 g (6.0 mmol) of triphenylphosphine and 5.35 mg (5 mmol) of 2,6-lutidine in 20 ml of dry dioxane , heated at 55 ° for 19 h and evaporated in vacuo. The dark red residue was chromatographed on a 35 g silica gel column. Elution with benzene ether gave 2.3 g (87%) of crude VII as a pink oil which was used without further purification in the next step.

^^ S-C0CH3 AgN03 /s._^,SAg ςΤ Β'-γ P «> 3 bases (Γ" y p03

COgPNB C02PNB

VII

X VIII

Mercaptide vm was prepared from VII according to the general procedure of Example III.

Cl \ N3 / Pyr ^ SAg 1) °

0 '' 'ys. P03 2) TFA

CVnb VIII 0 0 v »- ^ IX 'V» ΓΓ \ H2 / pd-c ^ J— N' - o OCHgCH2K3 x co2pnb ^^ Vch2och2ch2nh2

0 ^^ CO2H

XI

56 6 9 8 4 5

Reaction of mercaptide VIII and 2-azidoethoxyacetyl chloride according to the procedure of Example 1 gives intermediate IX, which can be cyclized and reduced as in Example 1 to give the title product.

Il

Claims (2)

57 6984 5 Claims approx A process for the preparation of antibacterial penem derivatives of the formula I and their pharmaceutically acceptable salts, is O, S or SO, Alk 'is C2-6 alkylene and Y is hydrogen or lower alkyl which may be substituted by hydroxy, characterized in that the compound of formula O -A- (Alk ') - N3 II CO.R "δ wherein Y, Alk, A and Alk' have the same meaning as above, R" is an easily cleavable ester-forming group and Q is phenyl or lower alkyl, cyclized in an inert organic solvent at a temperature slightly above room temperature and the reflux temperature of the solvent, after which the compound obtained is hydrogenated and, if desired, the compound of formula I obtained is converted into a pharmaceutically acceptable salt. Process for the preparation of 6- (1'-hydroxy-ethyl) -2- (2-aminoethoxymethyl) -penem-3-carboxylic acid according to Claim 1, characterized in that the starting material used is a compound of the formula II in which Y is CH 3 CH (OH) , Alk is CH2, A is O, Alk 'is CH2CH2, R "is p-nitrobenzyl and Q is phenyl.