DD247677A5 - PROCESS FOR THE PREPARATION OF 6- (SUBST.) - METHYLENE PENICILLAN- AND 6- (SUBST.) - HYDROXYMEHTYLPENICILLANIC ACID AND THEIR DERIVATIVES - Google Patents

Abstract

Process for the preparation of 6- (substituted) methylenpenicillan and 6- (subst.) Hydroxymethylpenicillanic acids and their derivatives. b-lactamase inhibiting compounds of the formulas or a pharmaceutically acceptable acid addition salt or carboxylate salt thereof, wherein n is 0, 1 or 2, X3 is H or Br, R1 is H, is the residue of certain carboxy protecting groups or the residue of an in vivo readily hydrolyzable ester group R12 and R13 is H and the other is vinyl, certain aryl, arylthio, alkylthio, alkylsulfonyl or certain heterocyclyl, aminomethyl, thiocarboxamido or amidino groups, one of R2 and R3 is H and the other such R18 is H or certain acyl groups, intermediates useful in their preparation, processes for their preparation and their use, and pharmaceutical compositions containing them. Formulas (I) and II)

Description

Process for the preparation of 6- (substituted) methylenpenicillan and 6- (substituted) hydroxymethylpenicillanic acid and their

derivatives

Field of application of the invention

The invention relates to novel 6- (substituted) -methylenepicillanic acids and novel 6- (substituted) hydroxymethylpenicillanic acids, certain esters and pharmaceutically acceptable salts thereof, pharmaceutical compositions containing them, processes for their preparation and their use as β-lactamase Inhibitors and intermediates for it.

Characteristic of the known technical solutions

One of the best known and most widely used classes of antibacterial agents is the class known as β-lactam antibiotics. These compounds are characterized by having a nucleus derived from a 2-azetidinone

20.1.83- 316U2 /,

(β-lactone) ring fused to either a thiazolidine or a dihydro-1,3-thiazine ring. When the nucleus contains a thiazolidine ring, the compounds are commonly referred to generically as penicillins, while when the nucleus is one Dihydrothiazine ring containing compounds called cephalosporins. Typical examples of penicillins commonly used in clinical practice are benzylpenicillin (penicillin G), phenoxymethylpenicillin (penicillin V), ampicillin and carbenicillin; Typical examples of common cephalosporins are cephalothin, cephalexin and cefazolin.

Object of the invention

However, despite the widespread use and widespread adoption of β-lactam antibiotics as valuable chemotherapeutic agents, they suffer the major drawback that certain anti-microorganism agents are not active. It is believed that in many instances this resistance of a particular microorganism to a given β-lactam antibiotic is due to the microorganism producing a β-lactamase. The latter substances are enzymes that cleave the β-lactam ring of penicillins and cephalosporins to products that have no antibacterial activity. However, certain substances have the ability to inhibit β-lactamase, and if a β-lactamase inhibitor is used in combination with a penicillin or cephalosporin, it may enhance or enhance the antibacterial efficacy of penicillin or cephalosporin against certain microorganisms. An enhancement of antibacterial activity is considered when the antibacterial activity of a combination of a β-lactamase inhibiting substance and a β-lactam antibiotic is substantially greater than the sum of the antibacterial activities of the individual components.

Explanation of the essence of the invention

Thus, according to the invention, novel compounds are provided which are 6- (substituted) - methylene penicillanic acids, their 1-oxides, 1,1-dioxides and in vivo readily hydrolyzable esters. These novel penicillanic acids and their in vivo readily hydrolyzable esters are potent inhibitors of microbial β-lactamases. Therefore, a method for enhancing the efficacy of β-lactam antibiotics using these new acids, their salts and certain readily hydrolyzable esters thereof is also provided.

Furthermore, derivatives of 6- (substituted) - methylenepenicillanic acids, their 1-oxides and 1,1-dioxides with a carboxy-protecting group are provided, these compounds being useful as chemical intermediates.

Still further provided are 6- (substituted) hydroxymethylpenicillanic acids, their 1-oxides, 1,1-dioxides, and salts and esters useful as both chemical intermediates and β-lactamase inhibitors.

European Patent Application No. 50,805 discloses compounds of the formula

⁽⁰⁾ n CH

</ ch ³

p ^CH 3 (III)

N _{/ y}

' ⁷ COOR ₃

wherein η is zero, 1 or 2, R is CN or certain carbonyl radicals, R _{2 is} hydrogen, lower alkyl or halogen and R _{3 is} hydrogen or a readily hydrolyzable group useful as β-lactamase inhibitors. The same reference discloses 6-oxopenicillanic acid esters, the corresponding sulfoxides and sulfones, and a process for their use.

316402

in the preparation of compounds of formula (III) by reaction with a phosphorane of the formula R 1 R ₂ C = P (C ₆ H ₅ J ₃ .

United Kingdom patent application GB 2 053 22OA discloses inter alia certain 6-methylene-1,1-dioxopenicillanic acids and esters of the above formula (III) wherein η is 2 and R and R independently are hydrogen, an optionally substituted alkyl, aryl -, optionally substituted cycloalkyl, an aralkyl or optionally substituted amino group or together with the carbon atom, y- .. 'to which they are attached, R ₁ and R- form a 3- to 7-membered carbocyclic or heterocyclic ring.

U.S. Patent 4,287,181 discloses certain 6-substituted penicillanic acid 1,1-dioxides and their esters wherein the 6-substituent OR ₃

R ₄ -CH

and R _{3 is} H or alkanoyl and R _{4 is} H, (C 1 -C ₄ ) alkyl, phenyl, benzyl or pyridyl, which are useful as β-lactamase inhibitors.

, The present invention provides new 6- (substituted) methylene

penicillanate of the formula

(0) "" _e

^CH3 (I) _f

O '''''' COOR ¹

wherein η is zero, 1 or 2, R a is ^a or R wherein R ^{a is} the residue of a carboxy-protecting group selected from tetrahydropyranyl, allyl, benzyl, 4-nitrobenzyl, benzhydryl, 2,2,2-trichloroethyl, t-butyl and phenacyl ; and R is hydrogen or the residue of an in vivo readily hydrolyzable ester group

is selected from 3-phthalidyl, 4-crotonolactonyl, γ-butyrolactone-4-yl ',

4 R ⁴ R ⁴ R ⁴

-CH -C = C-R

/ I, -COCOR, -COCOOR and -CHOCOR ¹⁴ ^ C- ⁰ R ⁵ . R ⁵

Il

and R and R are each hydrogen or CH ₃ , R is (C ₁ -C ₅ ) alkyl, and R ^{14 is}

CH,

or

where X "is the 3-substituent of a known cephalosporin

15 is β-lactam antibiotic, and R is the 6- or 7-substituent

a known penicillin or cephalosporin β-lactam

1 4 antibiotic is. Particularly preferred R are the above

1 5 penicillin residues, wherein R 2-phenylacetamido, 2-phenoxyacet-

amido, D-2-amino-2-phenylacetamido, D-2-amino-2- (4-hydroxyphenyl) acetamido, 2-carboxy-2-phenylacetamido, 2-carboxy-2- (2-thienyl) -acetamido ,. 2-carboxy-2- (3-thienylacetamido, D-2- (4-ethyl-2,3-dioxopiperazinocarbonylamino) -2-phenylacetamido, D-2- (4-ethyl-2,3-dioxopiperazinocarbonylamino) -2- (4-hydroxy-

phenyl) acetamido or 2,2-dimethyl-4-phenyl-5-imidazolidinone

2 is 3-yl; one of R and R is hydrogen and the other

Cl, CH ₂ OH, vinyl, (C, -C ₄₎ alkylthio, (C ₁ -C ₄₎ alkylsulfonyl, furyl, thienyl, N-methylpyrrolyl, N-Acetylpyrrolyl, RC, H, R ⁷ is C ^ -HS, - CH (R ⁴ JNR ¹⁶ R ¹⁷ , -CNR ⁸ R ⁹ , -CNR ⁸ R ⁹ ,

S NH

; ti ι. ο υ

3 -i 6 4 ü 2

⁴ JNR ¹⁶ R ¹⁷

R ⁷ C ₆ HS, -CH (R ⁴ JNR ¹⁶ R ¹⁷ , -CNR ⁸ R ⁹ , -CNR ⁸ R ⁹ ,

⁸ R ⁹

⁸ R ⁹

NH

(0)

N (O) 'P

CH.

(R ⁷ ) l ff

^NX n ^

N N

Il -h-

ν ^ n

• R

Ärtfc. , R ¹ Ip = ^ L, R ¹¹ s -ν

11 Ν "

and m is 2 or 3, ρ is zero or 1, t is zero, 1, or 2, X _{1 is} S, O or NR ¹¹ , R ^{7 is} hydrogen, (C ₁ -C ₄ ) alkyl, (C ₁ -C ₄ ) AIkoxy, allyloxy, hydroxyl, carboxyl, (C ₂ -C ₅ ) alkoxycarbonyl, (C ₁ -Ct) -alkylcarbonyl, phenyl, benzyl, naphthyl, pyridyl, NR R, CONR ⁰ R, NHCOR, NO ₉ , Cl , Br, CF ₃ or SR

on ^ J

R and R are each hydrogen, (C ₁ -C ₄ ) alkyl, phenyl or benzyl, R is (C ₁ -C ₄ ) alkyl, CF ₃ or phenyl,

• J ί.

-T-

R ^{11 is} hydrogen, CH ₃ , C ₃ H ₅ or CH ₃ CO, R and R are each H, (C.-C.) alkyl, (C ₃ -C.) Hydroxyalkyl, or taken together with the nitrogen atom on which they are hanging, R

and R are a 5- to 7-membered heterocyclic group, particularly preferred such heterocyclic groups are pyrrolidino, piperidino, morpholino, thiomorpholine or 4-methylpiperazino, or a pharmaceutically acceptable

2 3

an acid addition salt of the compound wherein R or R is a group containing a basic nitrogen atom or a pharmaceutically acceptable cationic salt of the compound wherein R is water; oxy! group included.

1 2 3

in which R is hydrogen or R or R is a carbohydrate

The above compounds wherein RR is ^a are useful as intermediates for the preparation of the compounds wherein R is R. The latter compounds are the active β-lactamase inhibitors of the invention.

12 13

The invention further provides 6- (R R -subst.) -Hydroxymethylpenicillanic acids, 1-oxides, 1,1-dioxides and esters thereof of the formula

- (II)

in which η and R are as described above for compounds of the formula (I)

* 12

X _{3 is} H or Br, one of the radicals R and R is hydrogen and the other vinyl, (C ₁ -C.) alkylsulfonyl, furyl, thienyl, N-methylpyrrolyl, N-acetylpyrrolyl,

9Ω Λ.85-

R ⁷ C, H ,, CH (R ⁴ JNR ¹⁶ R ¹⁷ , CNR ⁸ R ⁹ , CNR ⁸ R ⁹ ,

O ^, Il Il

S NH

R '

(Ο)

, R

(R ⁷ )

N.

Ν-N _R ll

"Ν-

N "11 S N" 11. 0

- -i- / λ 1- -Η- '"W- -j-,

N ^ N

or

12

provided that when R or R

N.

is and p

Zero, R has another meaning

18 ~

as H or CH ₃ , RH, (C ₃ -C ₅ ) alkanoyl, (C ₂ -C ₅ ) alkoxycarbonyl

yl, Pyraζincarbony1, benzoyl, CF ^ CO or C0NR ^ r9 is and

7 QQ Λ Λ Λ fi * Γ 7

m, ρ, t, R, R, R, R, R, R and X _{1 are} as defined above, or a pharmaceutically acceptable acid addition salt or carboxylate salt.

The compounds of formula (II) are all useful as chemical

· R

mixed intermediates for the preparation of the corresponding 6- (substituted) -methylerr-1, 1 -dxoxopenicillanate of the formula (I). In addition, however, the compounds of formula (II) wherein R 1 is hydrogen or the residue of an in vivo readily hydrolyzable ester group, R, as defined above, are useful because of their β-lactamase inhibitory activity, especially when used in conjunction with a β Lactam antibiotic used.

Particularly preferred compounds of the formula (I) are those

2 3 in which η is zero or 2 and one of the radicals R and R is hydrogen and the other furyl, thienyl, CH ₂ OH, phenyl, methylsulfonyl, N-methylpyrrolyl,

(R ⁷ ) HtT »(R ⁷ ) .H ^ l. _R Iin-? I,

, N

<* ⁷ > Ht if V

Particularly preferred compounds of the formula (II) are those in which η is zero or 2, one of the radicals R ₁₂ and R _{1 is} H and the other is vinyl, 2-furyl, 2-thienyl, N-methylpyrrol-2-yl, N Acetylpyrrol-2-yl, 3-hydroxy-2-pyridyl, 4-methoxy-2-pyridyl,

-ΙΟ

V-O3.

and R ^{18 is} H or CH ₃ CO.

Particularly preferred meanings for the carboxy-protecting group, R ^a , are allyl, benzyl, t-butyl and 2,2,2-trichloroethyl, and particularly preferred is allyl because of the relative ease with which it is selectively prepared and removed.

Particularly preferred as the residue of an in vivo readily hydrolyzable ester group, i. R are as defined above

_D 4 "4

-COCOR

or

-COCOOR ⁶

Il

4 5 and in particular those in which R and R are each hydrogen and R is as previously defined.

In addition to providing methods of preparation of the compounds of formulas (I) and (II), the invention further provides a method of treating a bacterial infection in a mammal, including humans, comprising administering an antibacterially effective amount of a compound of the formula (I ), where RR is as defined above

niert, to a sucker requiring such treatment comprises.

Also, pharmaceutical compositions are provided for treating a bacterial infection in mammals, including humans, which is an antibacterially effective amount.

effective amount of a compound of formula (I) wherein R is R

The compounds of formulas (I) and (II) wherein R is R as defined above are useful as inhibitors of β-lactamase enzymes. According to this mechanism, these compounds enhance the activity of β-lactam antibiotics (penicillins and cephalosporins), especially those microorganisms which are resistant to or resistant to the β-lactam antibiotic by the production of enzymes (β-lactamases) which otherwise destroy or partially destroy the β-lactam antibiotic. Thus, the spectrum of activity of the β-lactam antibiotic is increased.

Still further, the invention provides a method for treating a bacterial infection in a mammal, including a human, which comprises administering an antibacterially effective amount of a penicillin or cephalosporin, especially the above enumerated, and a β-lactamase inhibiting amount of a compound of the formula (FIG. I) or (II) to a mammal in need of such treatment.

While the compounds of the present invention are generally enhancing the activity of β-lactam antibiotics, their preferred use is in their combination with a penicillin or cephalosporin of established clinical utility, namely, amoxycillin, ampicillin, apalcillin, azlocillin, azthreonam, bacampicillin, carbenicillin, Carbenicillin indanyl, carbenicillin phenyl, cefaclor, cefadroxil, cefaloram, cefamandole, cefamandol nafate, cefaparol, cefatrizine, cefazolin, cefbuperazone, cefonicid,

ο Π 1 QS - ^ -i PA Γ ,?

£ 247 77

Cefmenoxime, Cefodizim, Cefoperazone, Ceforanide, Cefotaxime, Cefotiam, Cefoxitin, Cefpiramide, Cefpirome, Cefsulodin, Ceftazidime, Ceftizoxime, Ceftriazone, Cefuroxime, Cephacetril, Cephalexin, Cephaloglycine, Cephaloridine, Cephalothin, Cephapirin, Cephradine, Cyclacillin, Epicillin, Furazlocillin , Hetacillin, lenampicillin, levopropylcillin, mecillinam, mezlocillin, penicillin G, penicillin V, phenethicillin, piperacillin, pirbenicillin, pivampicillin, sarmoxicillin, sarpicillin, suncillin, talampicillin and ticarcillin, including their pharmaceutically acceptable salts. The terms used for these β-lactams are generally USAN, i. Terms used in the United States.

These also include combinations of the β-lactamase inhibitors of the invention with T- [1- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido., 3- (5,6-dihydro-4-pyrindenium) methyl-3 cephem-4-carboxylate (HR-810); 7 - / "2- (2-amino-4-thiazolyl) -2-methoxyiminoacetamido.7-3- (N-methylpyrrolidinium) methyl-3-cephem-4-carboxylate (BMY-28,142) and T- [D- ( 2 - / "4-carboxy-5-imidazolecarboxamido /) ^ -phenylacetamido. ZS - ^ - (2-sulfonato-ethyl) -pyridinum / 3-cephem-4-carboxylic acid.

Although the compounds of the invention may be administered separately from the β-lactam antibiotics, combination dosage forms are preferred. The pharmaceutical composition, whether for oral or parenteral use, comprises in a weight ratio of 1: 3 to 3: 1 a β-lactamase inhibitor of formula (I) or (II) and a β-lactam antibiotic in total amounts sufficient to successfully treat a bacterial infection in a nipple in a single or, more commonly, multiple doses.

The compounds of the formulas (I) and (II) according to the invention,

In which one of the groups R, R, R or R contains a basic nitrogen atom, acid addition salts are able to form.

3164G2

the. Such salts with pharmaceutically acceptable acids are within the scope of the invention. Examples of such acids are hydrochloric, hydrobromic, sulfuric, phosphoric, citric, maleic, tartaric, maleic, fumaric, gluconic, sugic, benzenesulfonic, p-toluenesulfonic, p-chlorobenzenesulfone and 2-naphthalenesulfonic acid.

Further, the compounds of formulas (I) and (II) wherein R is hydrogen form cationic salts and such salts with pharmaceutically acceptable cations are within the scope of the invention. Examples of such cations are sodium, potassium, ammonium, calcium, magnesium, zinc, and substituted ammonium salts formed with amines such as diethanolamine, choline, ethylenediamine, ethanolamine, N-methylglucamine and procaine.

The invention relates to derivatives of penicillanic acid by the following. Structural formula is reproduced:

In derivatives of penicillanic acid, a broken line bond (iJiHii) of a substituent on the bicyclic nucleus indicates that the substituent is below the level of the nucleus. Such a substituent is referred to as being in the <* configuration. Conversely, a broad line bond (- ^) of a substituent on the bicyclic nucleus indicates that the substituent is above the plane of the nucleus. This latter configuration is referred to as the β configuration. As

used here gives a solid line binding ()

of a substituent on the bicyclic nucleus, that the substituent may be either in the ε £ configuration or in the β configuration.

2 a 1.8 δ - 316432

The compounds of the invention are e.g. produced according to one or more of the following general methods.

Method A

-N.

.CH CH.

(IV)

'COOR

O O \ /

-N

(VII)

(VII)

= C ₆ H ₅

CH. CH.

^ COOR

-N-

CH ₃ CH ₃

⁷⁷ COOR ¹

(V)

3 ^ΘΘ RR ^J CP (0)

-N-

CH. CH.

COOR

(VI)

(VI)

methodB

0 \ / CH OH

"f ^ CH 1 -CH ₃ MgBr Ν L, 2.R ¹² R ¹³ C =

' ^R '

(Ix)

(VIII)

MethodeC

%, 2.R ^X 'R ^XJ C = O

'COOR

(II η = 2, X ₃ = H,

¹²

0 0

(II RVh, η = 2, X ₃ = R ¹⁸ = H)

0 0 ^Χ 'CH

CH ^

 Ν

(Viii)

br

O ^

(X)

'COOR

1 .CH ₃ MgBr or 13 tC ₄ H ₉ Li ^R OH I? (O) η CH ₃ .R ¹² R ¹³ C = O Br J 2 L I ** 3 ^ COOR ¹ -N-

, n = 0 or (XI) R ¹

18

OR R ¹³ -C.

R ¹²

CH.

-N-

ViH f <NC ™

(II, X ₃ = H)

^ COOR ¹

-Br -N

(XII)

(I) Λ Λ Λ

-1 U, / J ^ "

The required Wittiq salts of the formula R ² R ³ CHI ^ (CeH.-)., Cl ⁶ , which are used in the above method A, are either known compounds or are prepared from commercially available precursors by customary synthesis methods, as illustrated below ,

Production of Wittig salts

CH ₃ OH

^Na 2 ^S 2 ° 8

SOC1_P (0), £ -> i.

^ OH

(CH ₃ CO)

Cl CH ₃ Cl 1 CH ₃ .

't

P0 ₃ C1

Cl CH ₂ OCOCH ₃

Cl

0 ₃ P Cl

CH

Cl

The primary alcohols of the general formula R CH ₂ OH are converted to the corresponding chloromethyl compounds, typically by reacting the alcohol with an equimolar amount of thionyl chloride in the presence of a reaction inert solvent, eg chloroform or methylene chloride, at or about room temperature. The product is isolated, for example by neutralizing the reaction mixture and extracting.

The chloromethyl compound, R CH ₂ Cl, is then converted to the desired Wittig salt, for example by reaction with an equimolar amount of triphenylphosphine. Typically, this step occurs in a solvent such as toluene at elevated temperature, preferably at the reflux temperature. The desired product forms a precipitate, which is then collected by filtration.

6- O-hydroxypenicillanic acid is a known compound, see, for example, Hauser et al., HeIv. Chim. Acta, J50, 1327 (1967). The acid is converted to a carboxy-protected derivative of formula (IV). The identity of the carboxy-protecting group is not critical. The only requirements for the carboxy-protection. group R ^a are that (i) they must be stable to the oxidation conditions used to form the 6-0xopenicillanate ester (V) and the subsequent reaction with the Wittig reagent to form the 6- (subst.) Methylene penicillanate of the formula (VI, R = R ^a ), (ii) they are selective

1 a must be removable from the compound of formula (VI, R = R) using conditions under which both the β-lactam and the 6- (substituted) methylene groups remain virtually intact, (iii) they are resistant to oxidation the compound (VI, R = R ^a ) must be stable to the sulfones of the formula (VII) or the corresponding sulfoxide oxides. Such typical carboxy-protecting groups meeting the above requirements are the tetrahydropyranyl group, the benzyl group, the benzhydryl group, 2,2,2-trichloroethyl group, the allyl group, the t-butyl group and the phenacyl -Group. See also US-PS

3,632,850 and 3,197,466, British Patent 1,041,985, Woodward et al., Journal of the American Chemical Society, 8J5, 852 (1966), Chauvette, Journal of Organic Chemistry, 3, 1259 (1971), Sheehan et al., Journal of Organic Chemistry, 2_9, 2006 (1964), and "Cephalosporin and Penicillin, Chemistry and Biology", eds. HE Flynn, Academic Press, Inc., 1972. Such particularly preferred groups are allyl, benzyl, and 2, 2,2-trichloroethyl and especially preferred is allyl due to its ease of preparation and selective removal.

L. / The oxidation of the carboxy-protected 6-o-hydroxypenicillanate (IV) to the corresponding 6-oxo-penicillanate ester (X) is typically carried out with an approximately equimolar amount of trifluoroacetic anhydride and a molar excess of dimethyl sulfoxide in the presence of a reaction inert Solvent, eg Chloroform or methylene chloride. The reaction is preferably carried out at a temperature of about -80 to -70 ° C. The reaction mixture is neutralized, e.g. by adding a tertiary amine such as triethylamine, whereupon the mixture is isolated, e.g. by partitioning between water and a water-immiscible solvent and evaporating the organic layer.

The 6-Oxopenicillanatester of formula (V) is then with a

2 3 ^Θ ®

Wittig reagent of the formula RR CP (CgH ₅ ) ₃ implemented. This reaction is preferably carried out in the presence of a reaction-inert organic solvent, for example a hydrocarbon such as pentane, hexane, benzene, toluene or xylene, a halogenated hydrocarbon such as methylene chloride, chloroform, carbon tetrachloride, 1,2-dichloroethane, 1,2-dibromoethane or chlorobenzene , an ether such as tetrahydrofuran, dioxane, diethyl ether, 1,2-dimethoxyethane or t-butyl ethyl ether. While this reaction can be over a temperature range from about -100 to +50 ⁰ C., a preferred temperature ranging from about -78 to 25 C.

0 1.85- 3 "164Ul

9 0

The desired product of formula (VI, R = R ^a ) is isolated by known techniques, eg the reaction is quenched by the addition of aqueous ammonium chloride, extracted with a water-immiscible solvent and the solvent evaporated. The resulting product is purified, if desired, by conventional methods known to those skilled in the art, for example by column chromatography on silica gel.

1a a The esters of formula (VI, R = R), wherein R is a carboxy-protecting group as defined above, can then be converted to the corresponding acid or ester of formula (VI, R = R) wherein R is hydrogen or a is an in vivo easily hydrolyzable ester-forming radical. Typically, the carboxy-protecting group is cleaved from the intermediate compound of formula (VI, R = R ^a ) to form the corresponding carboxylic acid. The particular method chosen to remove the carboxy protecting group will depend on the exact nature of the ester residue R ^a , but a suitable method will be readily apparent to one skilled in the art.

As mentioned above, a particularly preferred carboxy-protecting group R is allyl. While this group can be cleaved by mildly acidic or alkaline hydrolysis procedures with satisfactory results, a particularly preferred method of their cleavage is the use of a soluble palladium (0) complex, tetrakis (triphenylphosphine) palladium (0) as a catalyst, previously described by Jeffrey and McCombie, J. Org. Chem., £ 7, 587-590 (1982). In a typical procedure, the allyl ester is dissolved in reaction inert solvent, e.g. Ethylene dichloride, chloroform, ethyl acetate, and a catalytic amount of tetrakis (triphenylphosphine) palladium (0), e.g. about 1 to 5 mol%, based on the allyl ester, and an approximately equal weight of tri-phenylphosphine brought together under a nitrogen atmosphere. For this purpose, a sodium or potassium salt of 2-ethylhexanoate is added in an equimolar amount to the starting allyl ester, and

20Λ8-5- S-i 643

Oji

the resulting mixture is stirred at room temperature until the desired salt, e.g. of the formula (VI), wherein R is Na or K, has completely precipitated. Usually, the reaction is practically complete in about two to twenty hours. The salt is then collected, e.g. by filtration.

When sulfoxides or sulfones according to the invention are desired, for example those of formula (I) wherein η is 1 or 2, the sulfides of formula (VI) are oxidized using any of a wide variety of oxidizing agents known in the art the oxidation of sulfoxides to sulfones is known is used. However, particularly convenient reagents are metal permanganates, such as the alkali metal permanganates and alkaline earth metal permanganates, and organic peroxyacids, such as organic peroxycarboxylic acids. Suitable single reagents are sodium permanganate, potassium permanganate, 3-chloroperbenzoic acid and peracetic acid.

A particularly preferred group of oxidizing agents are the organic peroxyacids, and a preferred organic peroxyacid is 3-chloroperbenzoic acid.

When the desired oxidation product is a sulfoxide of formula (I) wherein η is 1, approximately molar equivalents of the starting sulfide (n = 0) and the oxidizing agent are employed. When the desired product is a sulfone, e.g. of the formula (I) wherein η is 2, the sulfide is contacted with two or more mole equivalents of oxidizing agent. On the other hand, of course, the sulfoxides can serve as starting materials for the preparation of the corresponding sulfone, in which case at least about an equimolar amount of oxidizing agent is used.

For example, when a compound of the formula (VI> R = R ^a ) in which R ^{a is} as previously defined is oxidized to the corresponding compound of formula (VII) using an organic peroxy acid, eg a peroxycarboxylic acid, the

Reaction usually carried out by treating the compound of formula (VI, R = R) with about two to about four molar equivalents of the oxidizing agent in a reaction-inert organic solvent. Typical solvents are chlorinated hydrocarbons such as dichloromethane, chloroform and 1,2-dichloroethane, and ethers such as diethyl ether, tetrahydrofuran and 1,2-dimethoxyethane. The reaction is usually preferably carried out at a temperature of about -20 to about 50 ⁰ C and at about 25 ° C. At about 25 ° C, reaction times of about 2 to about 16 hours are usually used. The product is usually isolated by removing the solvent by evaporation in vacuo. The product can be purified by conventional methods well known in the art.

In the above-mentioned oxidation procedures, it is preferred to use a starting material in which the carboxy group is protected by the above-mentioned carboxy-protecting groups, R '. Cleavage of the carboxy-protecting group from the product sulfoxide or sulfone is normally effected for the particular protecting group used, e.g., as described above for the compounds (VI, R = R).

The compounds of the invention, e.g. of formula (I) or (II) wherein R is an in vivo readily hydrolyzable ester-forming group can be prepared directly from the corresponding compound wherein R is hydrogen by conventional esterification techniques. The particular method chosen will depend on the exact structure of the ester-forming moiety, but a suitable method will be readily selected by those skilled in the art. In the event that R is 3-phthalidyl, 4-crotonolactonyl, y-butyrolactone-4-yl and groups of the formulas

R ⁴ f

-COCOR ⁶ and -COCOOR ⁶

ο.-; c λ

247 67?

wherein R, R and R are as defined above, they may be prepared by alkylating the appropriate compound of the invention wherein R is hydrogen with a halide of the formula RQ, ie, a 3-phthalidyl halide, a 4-crotonolactonyl halide, a y- Butyrolactone-4-yl halide or a compound of the formula

4 4

R R

ιc ig

/ \, QCOCOR or QCOCOOR

V * ⁵

wherein Q is halogen and R, R and R are as previously defined. The terms "halide" and "halogen" are intended to mean derivatives of chlorine, bromine and iodine. The reaction is typically carried out by dissolving a salt of the compound e.g. of the formula (I) or (II) wherein R is hydrogen in a suitable polar organic solvent, e.g. Ν, Ν-dimethylformamide, and then adding about one molar equivalent of the appropriate halide (R Q). When the reaction is almost complete, the product is isolated by standard techniques. Often it is sufficient simply to dilute the reaction medium with an excess of water and then extract the product into a water-immiscible organic solvent and then recover it by solvent evaporation. Salts of the starting materials which are commonly used are alkali metal salts / such as sodium and potassium salts, tertiary amine salts such as triethylamine, N-ethylpiperidine, N, N-dimethylaniline and N-methylmorpholine salts and. quaternary ammonium salts, such as tetramethylammonium and tetrabutylammonium salts. The reaction is conducted at a temperature in the range of about 0 to 100 ° C and usually at about 25 ° C. The length of time needed to achieve full implementation varies with a variety of factors, such as

4 ** / W f #

Concentration of the reaction components and the reactivity of the reagents. Thus, considering the halogen compound, the iodide reacts faster than the bromide, which in turn reacts faster than the chloride. In fact, it is sometimes advantageous to add up to one mole equivalent of an alkali metal iodide when using a chlorine compound. This has the effect of speeding up the reaction. Taking full account of the above factors, reaction times of about 1 to about 24 hours are usually employed.

If method B, as stated above, for the preparation of the compounds of formula (II) according to the invention, wherein η 2 and

1 8 X, and R are each hydrogen, or compounds of formula (VIII) is employed, the required 6-oG-bromo-1,1-dioxopenicillanate ester starting material (IX) is converted into a Grignard reagent by reacting with an equimolar amount of a low molecular weight Grignard reagent, eg Ethyl magnesium bromide, ethyl magnesium chloride or n-butyl magnesium iodide, in ethereal solvent, preferably tetrahydrofuran or ethyl ether, at a temperature of -80 to 25 ° C, typically -78 ° C. After a few minutes of stirring, an equimolar amount of the appropriate aldehyde is added

12 13 of the formula R R C = O is added and stirring is continued until the reaction is virtually complete, usually for about 10 minutes to about 4 hours at the same temperature. The desired ester of formula (II), n = 2, is then isolated by standard methods. For example, the reaction is quenched with aqueous ammonium chloride and the product extracted with a water-immiscible solvent. The resulting product (II) is further purified, e.g. by silica gel chromatography.

The secondary alcohol of formula (II), n = 2, X ₃ = H, can then be dehydrated to yield the corresponding 6- (substituted) methylene-1, 1-dioxopenicillanate compound of formula (VIII). While a variety of known in the art for the dehydration of secondary alcohols to olefins

Methods can be successfully applied to carry out this step, a preferred method uses the conversion of the alcohol into an acetate by reaction with at least äguimolaren amounts of acetic anhydride and pyridine and then stirring at room temperature for 1 to 10 hours to form the olefin in substantial quantities. The reaction is usually quenched with water and the desired product (II), n = 2, is isolated by extraction methods and purified if desired.

The products of formula (II) or (VIII) obtained as described above are esters in which R is either a carboxy-protecting group, R ^a , as defined above, or the residue of an in vivo readily hydrolyzable ester group, R, as defined above, is. These esters, wherein ^A is RR are converted into the corresponding carboxylic acids (R = hydrogen) by methods described above. Of course, if desired, the carboxylic acids of formulas (II) and (VIII) are converted to a corresponding compound wherein r 'is the residue of an in vivo readily hydrolyzable ester group by methods also described above.

Typically, the starting 6- U- bromo-1,1-dioxopenicillanate esters (IX) are prepared from 6,6-dibromo-1,1-dioxopenicillanic acid by treatment with sodium bicarbonate and sodium bisulfite followed by acidification. The resulting 6-i-bromo-1, 1-dioxo-penicillanic acid is then converted into an ester of the formula (IX).

The starting esters of formula (X) employed in method C, as set forth above, are known compounds, see e.g. U.S. Patent 4,234,579. In a typical procedure carried out by this method, the starting ester (X) is dissolved in reaction inert solvent, e.g. Toluene, xylene, pentane, tetrahydrofuran, ethyl ether or mixtures thereof, at low temperature with an equimolar amount of alkyllithium reagent, e.g. n-butyllithium, t-butyl, lithium or methyl

lithium, brought together to form a lithiopenicillin intermediate. This is immediately measured with an equimolar

12 13 1213

aldehyde, R ^A R CO, in which RR are as defined above, and the mixture is stirred for about 1 to 4 hours at -100 to -50 ° C, preferably at -78 ° C. The reaction is then quenched and the bromohydrin intermediate of formula (XI) isolated, for example by partitioning between water and solvent and purifying the extract by column chromatography on silica gel or florisil (magnesium silicate).

Alternatively, the above starting dibromoester of formula (X) is reacted with an equimolar amount of a low molecular weight Grignard reagent using the same reagents and conditions described above for Method B to provide the bromohydrin of formula (XI).

The bromohydrin (XI) can be converted into the corresponding compound (XII)

1 8

be acylated, wherein R has a meaning other than hydrogen, as defined above. Typically, the acylation is accomplished by reacting equimolar amounts of acyl chloride, acyl bromide or the corresponding acid anhydride, the intermediate ene-bromohydrin of formula (XI) and a tertiary amine, e.g. Pyridine, N-methylmorpholine or the like, in the presence of a reaction-inert organic solvent, preferably methylene chloride, tetrahydrofuran or ethyl acetate, at or below room temperature. The desired diester of formula (XII) is then isolated by well-known methods such as extraction and evaporation of the solvent and, if desired, purified, e.g. by column chromatography.

The bromohydrin ester intermediate (XI) or bromo diester (XII) may then be subjected to hydrogenolysis conditions to cleave the bromine atom. This is done by employing any one of a variety of known reducing agents and conditions, e.g. by exposing the bromohydrin

M "1.86-" 3 ί 64-32

_ 26 -

Hydrogen in the presence of a noble catalyst or reduction by means of certain organotin hydrides.

Preferred organotin hydride reducing agents are the dialkyltin dihydrides, trialkyltin hydrides having from 1 to 6 carbon atoms in each of the alkyl groups, and the triaryltin hydrides wherein the aryl is phenyl or phenyl substituted by nitro or alkyl or alkoxy having from 1 to 3 carbon atoms. Particularly preferred are triphenyltin hydride and tri-n-butyltin hydride, the latter being particularly preferred for reasons of economy and performance.

The reaction carried out using the tin hydrides is usually carried out in the presence of a reaction-inert solvent. Suitable solvents for use with the organotin hydride reducing agents are those which substantially dissolve the starting compound of formula (XI) or (XII) but do not themselves react with the hydride reducing agent. Examples of such solvents include the aromatic hydrocarbons, such as benzene, toluene, xylene, chlorobenzene and naphthalene, and ethers, such as ethyl ether, isopropyl ether, tetrahydrofuran, dioxane and 1,2-dimethoxyethane. Particularly preferred solvents for reasons of economy and performance are benzene and toluene.

In carrying out the hydrogenolysis using organotin hydride reducing agents, theoretically equimolar amounts of bromohydrin (XI) or bromo diester (XII) and hydride are required. In practice, an excess of hydride, e.g. 5 to 50 mole percent excess applied to ensure complete reaction.

Hydrogenolysis by organotin hydrides is virtually complete under the preferred conditions disclosed above without the use of a catalyst. However, the reaction is accomplished by a source of free radicals, e.g. UV-

LT

Light, or a catalytic amount of azobisisobutyronitrile or peroxides, such as benzoyl peroxide accelerated. A catalytic amount of azobisisobutyronitrile is a preferred source of free radicals for this reaction.

Typically, the compound of formula (XI) or (XII) is dissolved in reaction inert solvents, the solution is maintained under an inert atmosphere, eg, a nitrogen or an argon atmosphere, and the appropriate amount of organotin hydride and optionally the free radical source, For example, azobisisobutyronitrile, added and the resulting mixture is stirred at a temperature in the preferred range of about ^{0 0} C to the boiling point of the solvent. The reaction is usually completed in a few minutes to about a few hours, for example, from 5 minutes at the benzene boiling point to about 2 hours at ⁰ ° C. The product of formula (II, X-, = H) is then isolated by methods known to those skilled in the art. For example, by evaporation of the solvent and silica gel chromatography of the residue.

The compounds of formula (II, X-, = H) formed by organotin hydride debromination as described above

12 13 are predominantly 6-β isomers, that is, the 6-R R -

1 8 C (OR) substituent is in β configuration.

When the hydrogenolysis is carried out using hydrogen in the presence of a noble metal catalyst, there is a convenient method of carrying out this transformation by stirring or shaking a solution of a compound of the formula (XI) or (XII) under an atmosphere of hydrogen or hydrogen in admixture with one inert diluents, such as nitrogen or argon, in the presence of a noble metal hydrogenolysis catalyst. Suitable solvents for this hydrogenolysis reaction are those which practically dissolve the starting compound of the formula (XI) or (XII) but which themselves do not undergo hydrogenation or hydrogenolysis. examples

2a Ι.8δ- 316432

Examples of such solvents include ethers such as diethyl ether, tetrahydrofuran, dioxane and 1,2-dimethoxyethane, low molecular weight esters such as ethyl acetate and butyl acetate, tertiary amides such as N, N-dimethylformamide, Ν, Ν-dimethylacetamide and N-methylpyrrolidone, water and Mixtures of this. In addition, it is often desirable to buffer the reaction mixture to operate at a pH in the range of about 4 to 9, and preferably about 6 to 8. Borate, bicarbonate and phosphate buffers are commonly used. The introduction of the hydrogen gas into the reaction medium is usually carried out by carrying out the reaction in a closed vessel containing the compound of the formula (XI) or (XII), the solvent, the catalyst and the hydrogen. The pressure in the reaction vessel may vary from about 1 to about 100 bar (kg / cm ² ). The preferred pressure range when the atmosphere in the reaction vessel is substantially pure hydrogen is about 2 to about 5 bar (kg / cm ² ). The hydrogenolysis is generally carried out at a temperature from about 0 to about 60 ° C, and preferably from about 25 to about 50 ° C. When using the preferred temperature and pressure values, the hydrogenolysis generally takes place in a few hours, for example about 2 to about 2 hours. The preferred noble metal catalysts used in this hydrogenolysis reaction are the type of agents known in the art for this type of conversion, eg, nickel, palladium, platinum and rhodium. Palladium is particularly preferred. The catalyst is usually present in an amount of from 0.1 to about 25 weight percent, and preferably from about 0.1 to about 10 weight percent, based on the compound of formula (XI). It is often convenient to suspend the catalyst on an inert support; a particularly convenient catalyst is palladium suspended on an inert support such as carbon.

When the hydrogenolysis is virtually complete, the desired product of formula (II, X ^ = H) is then isolated by standard methods, e.g. the catalyst is filtered by

2a 1.85- 3-i64ü2

The solvent is evaporated and the product, if desired, purified by known methods, such as crystallization or by chromatography.

When the starting compound of the formula (XI) or (XII) is a benzyl ester (R = R ^a = benzyl), the above catalytic hydrogenolysis work may also cause cleavage of the benzyl group / and the product is of the formula (II) wherein X is ₃ and R are each hydrogen.

The 6- (substituted) hydroxymethylpenicillanic acid or ester of formula (XII) or (II, Xo = H) wherein η is zero can be oxidized by any of the methods known for the conversion of sulfides to sulfoxides and sulfones, eg with the aid of 3-chloroperbenzoic acid as described above to afford the corresponding sulfoxide or sulfone of formula (XII) OR (II, X2 = H) and where in each η is 1 or 2, respectively. However, a preferred method for obtaining sulfones of formula (XI), (XII) or (II) is to use the appropriate 6,6-dibromo-1,1-dioxopenicillanate ester (X), where η = 2, as the starting material in the Use method C described above.

1213 12 13

The starting aldehydes of the formula R R CO, wherein R and R are as defined above, are either available from a commercial source or are readily prepared from available starting materials by methods well known in the art, e.g.

1. Oxidation of the corresponding primary alcohols provided above as Wittig reagent precursors using e.g. of oxidants, such as potassium dichromate, chromic acid / pyridine, catalytic oxidation in the presence of precious metals, manganese dioxide.

2. Reaction of the corresponding methyl-substituted aromatic hydrocarbon with e.g. Selenium dioxide.

3-1643 *

3. Metal hydride reduction of the corresponding Cj-C.-alkoxycarbonyl compound at low temperature in the presence of ethereal solvents. Examples of suitable metal hydrides are lithium aluminum hydride and diisobutylaluminum hydride (DIBAL-H).

4. Reaction of a suitable aromatic hydrocarbon precursor with n-butyllithium and dimethylformamide.

As indicated above, the compounds of formulas (I) or (II) wherein R is H and their salts in combination with β-lactam antibiotics exhibit synergistic activity in in vitro antibacterial assays. Such activity is demonstrated by measuring the minimum inhibitory concentrations (MIC) in μg / mL against a variety of microorganisms. The procedure followed is the brain heart infusion (BHI) recommended and used by the International Collaborative Study on Antibiotic Sensitivity Testing (Ericcson and Sherris, Acta Pathologica et Microbiologia Scandinav, Supp. 217, Section B: 64-68 (1971)). Agar and the inocula replicating device. Overnight growth tubes are diluted 100-fold for use as a standard inoculum (20,000 to 10,000 cells in approximately 0.002 ml are placed on the agar surface, 20 ml BHI agar / plate). 12 2-fold dilutions of the test compound are used, with the starting concentration of the test drug being 200 μg / ml. Single colonies are discarded when plates are read after 18h at 37 ° C. The susceptibility (MIC) of the test organism is assumed to be the lowest concentration of the test compound or combination of compounds capable of producing complete growth inhibition as judged by the naked eye.

Such compounds of formulas (I) and (II) wherein R is H, and their salts in combination with known β-lactam antibiotics are industrial antimicrobial agents

, 1 w o.?.

έ 4 / O / /

usable, e.g. in water treatment, sludge control, color preservation and wood preservation, as well as for topical use as a disinfectant. In the case of using these compounds for such an application, it is often convenient to mix the active ingredient with a non-toxic carrier, such as a vegetable or mineral oil, or a softening cream. Likewise, they can be dissolved or dispersed in liquid diluents or solvents, such as water, alkanols, glycols or mixtures thereof. In most cases, it is appropriate to use active ingredient concentrations of from about 0.1 to about 10 percent by weight, based on the total composition.

As also noted above, the compounds of formulas (I) and (II) wherein R is R are of particular value as potent inhibitors of m -robacterial β-lactamases. By this mechanism they enhance the antibacterial efficacy of β-lactam antibiotics (penicillins and cephalosporins) over many microorganisms, especially those that produce β-lactamase. The ability of the compounds of formulas (I) or (II) to increase the efficacy of a β-lactam antibiotic can be estimated by reference to experiments in which the MIC values of the antibiotic alone and a compound of formula (I) or (II) be determined with R = hydrogen alone. These MIC values are then compared with the MIC values obtained with a combination of the given antibiotic and the compound of formula (I) or (II) wherein R = hydrogen. If the antibacterial strength of the combination is substantially greater than would be predicted from the strengths of the individual compounds, this is considered to be an enhancement of activity. The MIC values of combinations are determined using Barry and Sabath in "Manual of Clinical Microbiolgy", edited by Lenette, Spaulding and Truant, 2nd edition, 1974, American

Society for Microbiology / method described.

The compounds of formulas (I) and (II) wherein R is hydrogen or the residue of an in vivo readily hydrolyzable ester group enhance the antibacterial activity of β-lactam antibiotics in vivo. That is, they lower the amount of antibiotic needed to protect mice against otherwise lethal inoculation with certain β-lactamase producing bacteria. In determining such activity, acute experimental infections in mice are caused by intraperitoneal inoculation of the mice with a standardized culture of the test organism suspended in 5% porcine stomach mucin. The severity of the infection is standardized so that the mice receive a lethal dose of the organism (the lethal dose is the minimum inoculum of the organism required to reliably kill 100% of the infected untreated control mice). The test compound in combination with antibiotic is administered in various dosage amounts, po or ip, to groups of infected mice. At the end of the test, the activity of the mixture is determined by counting the number of survivors among treated animals at a given dose. The activity is expressed as percentage of animals which survive at a given dose, or calculated as PD_ _n (dose which protects 50% of the animals from infection).

The. The ability of the compounds of formulas (I) and (II) to enhance the efficacy of a β-lactam antibiotic against β-lactamase producing bacteria makes them valuable for coadministration with β-lactam antibiotics in the treatment of bacterial infections in mammals, particularly humans. In the treatment of a bacterial infection, the compound of formula (I) or (II) may be mixed together with the β-lactam antibiotic, or in the case where R is CH (R) OCOR, where R and R ^{14 are} as defined above the β-lactam antibiotic to the compounds

sw »-» »'τ β ig

compound of formula (I) or (II) chemically bound, and the two agents are thereby administered simultaneously. Alternatively, the compound (I) or (II) may be administered as a separate agent in the course of treatment with a β-lactam antibiotic. In some cases, it is advantageous to pre-dose the patient with the compound of formula (I) or (II) before starting treatment with a β-lactam antibiotic.

When a compound of formula (I) or (II) wherein R is R, as defined above, is used to enhance the activity of a β-lactam antibiotic, a mixture of the compound with the β-lactam antibiotic or the

b 4 to the compound of the invention alone when R is CH (R) OCOR, preferably administered in formulation with standard pharmaceutical carriers or diluents. A pharmaceutical composition comprising a pharmaceutically acceptable carrier, _a: comprises ß-LactamantibiotLkum and / or the compound of formula (I) or (II) will normally contain from about 5 to about 80 wt .-% of the pharmaceutically acceptable carrier.

When the compounds of formula (I) or (II) are used in combination with another β-lactam antibiotic, the. Compounds oral or parenteral, i. be administered intramuscularly, subcutaneously or intraperitoneally. Although the prescribing physician will ultimately decide on the dosage to be applied to a human patient, the ratio of the daily dosages of the compound of formula (I) or (II) wherein R is R and the β-lactam antibiotic will normally be in the range of about 1: 3 up to 3: 1 on a weight basis. In addition, when the compounds of formula (I) or (II) are used in combination with a β-lactam antibiotic, the daily oral dosage of each component is usually in the range of about 10 to about 200 mg / kg body weight and the daily Parenteral dosage of each component will normally be from about 10 to about 40 mg / kg of body weight.

These daily doses are usually divided. In some cases, the prescriber will determine that dosages outside these limits are needed.

As those skilled in the art will recognize, some β-lactam compounds are effective when administered orally or parenterally while others are only effective when administered parenterally. When a compound of formula (I) or (II) is to be used concomitantly (i.e., mixed) with a β-lactam antibiotic which is effective only on parenteral administration, a combination formulation suitable for parenteral use is necessary. When a compound of formula (I) or (II) is to be used simultaneously (mixed together) with a β-lactam antibiotic which is orally or parenterally active, combinations suitable for either oral or parenteral administration can be prepared. In addition, it is possible to orally administer preparations of the active compounds of the formula (I) or (II), while at the same time a further β-lactam antibiotic is administered parenterally; and it is also possible to administer preparations of the compounds of formula (I) or (II) parenterally, while at the same time orally administering the further β-lactam antibiotic.

The present invention is illustrated by the following examples. It should be understood, however, that the invention is not limited to the specific details of these examples. Proton and C nuclear magnetic resonance spectra were measured at 60, 90, 250 or 300 MHz for solutions in deuterochloroform (CDCl ₃ ), · deuterium oxide (D-0), perdeuteroacetone (CD ₃ COCD ₃ ) or perdeuterodimethylsulfoxide (DMSO-d _g ) , and peak levels are expressed in parts per million (ppm) downfield of tetramethylsilane. The following abbreviations are used: s = singlet; d = doublet; dd = doublet of doublets; t = triplet; q = quadruplet;

m = multiplet; b = wide. EXAMPLES Example 1 6-pi-hydroxypenicillanate ester

A. Allyl ester

A solution of 85 g of 6- * 6-hydroxypenicillanic acid (prepared according to the method of Hauser et al., HeIv. Chim. Acta. 50 , 1327 (1967), 0.39 mol) in 300 ml of dimethylformamide was mixed with 34 ml (0 , 39 mol) of allyl bromide, 54 ml (0.39 mol) of triethylamine and 2 g of sodium bicarbonate and the mixture stirred for 15 h at room temperature. After quenching the reaction with water and extracting with ethyl ether, the combined ether layers were washed with saturated sodium bicarbonate solution, water, dried (over MgSO 4), and concentrated in vacuo to 43 g of crude product. The crude material was purified by silica gel column chromatography eluting with 9: 1 chloroform / ethyl acetate to give 22.75 g (23%) of the allyl ester. ¹ H-NMR _(CDCl3) ppm (£): 1.42 (s, 3H), 1.60 (s, 3H), 4.45 (s ,. 1H), 4.5-5.0 (m , 3H), 5.2-6.2 (m, 4H).

B. pivaloyloxymethyl ester

A mixture of 9 g (0.041 mol) of 6- "f -hydroxypenicillanic acid, 40 ml of dimethylformamide, 7.4 ml (0.041 mol) of diisopropylethylamine, 6 ml (0.041 mol) of chloromethyl pivalate, and 6.15 g (0.041 mol) of sodium iodide was added at room temperature stirred for 15h. Water was added, the mixture extracted with ethyl ether, the extracts dried and concentrated to give 9 g of crude ester, which was purified on a silica gel column eluting with chloroform / ethyl acetate (9: 1). The combined product fractions amounted to 4.384 g

(32%).

£ * i / QI £

C. Benzyl ester

To a mixture of 20 g (0.092 mol) 6-t / hydroxypenicillanic acid, 12.9 mL (0.092 mol) triethylamine, 1.105 g (0.013 mol) sodium bicarbonate and 200 mL dimethylformamide (DMF) was added 12.0 mL (0.110 mol ) Benzyl bromide. The mixture was stirred at room temperature for 20 h, partitioned between ethyl ether and water and the aqueous phase adjusted to pH 2.0 with 6N hydrochloric acid. The layers were separated, the aqueous layer extracted twice with ether, the combined ether layers washed with sodium bicarbonate solution, water, dried and the solvent evaporated. The residue was crystallized from hot chloroform / hexane to give 9.1 g of colorless crystals, m.p. 165-167 ° C.

D. (5-Methyl-2-oxo-1,3-dioxol-4-yl) methyl ester

A mixture of 1.5 g (0.078 mol) of (5-methyl-2-oxo-1,3-dioxol-4-yl) methyl bromide, 18.7 g (0.078 mol) of sodium 6- hydroxypenicillanate in 225 ml of DMF becomes 4 Stirred at room temperature, poured into ice and worked up as described above to provide the desired ester.

Example 2

6-Oxopenicillanatester

A. allyl 6-oxopenicillanate

A mixture of 2.84 ml of [0.04 mol] dimethylsulfoxide, 3.67 ml (0.026 mol) of trifluoroacetic anhydride and 50 ml of methylene chloride was stirred at -78 ° C for 10 min. A solution of 5.14 g 0.02 mol) of allyl 6-IKr hydroxypenicillanat in 10 ml of methylene chloride was added at -78 ⁰ C and the resulting mixture stirred for 40 min. Triethylamine (7.24 ml, 0.052 mol) was added at this temperature and the mixture was gradually warmed to room temperature and quenched with water. After this

Extract with methylene chloride, wash the combined organic layers with water (3x), dry, and evaporate the solvent in vacuo to give the title compound, a yellow oil, 5.1 g (100%). H-NMR _(CDCl3) ppm (£): 1.60 (s, 6H), 4.75 (m, 2H), 4.82 (s, 1H), 5.1 to 6.3 (m, 3H ), 5.82 (s, 1H).

B. pivaloyloxymethyl ester

A mixture of 0.36 ml (5.06 mmol) of dimethylsulfoxide, 0.47 ml (3.29 mmol) of trifluoroacetic anhydride, 839 mg (2.53 mmol) of pivaloyloxymethyl-6-hydroxybuticillanate and 5 ml of methylene chloride was added for 30 minutes Stirred to -78 ° C, and 0.92 ml (6.58 mmol) of triethylamine were added. Workup of the product as described in Part A, above, gave 788 mg (95%) of the desired ketone. ¹ H-NMR _(CDCl3) ppm (£): 1.3 (s, 9H), 1.65 (s, 6H), 4.85 (s, 1H), 5.8 (m, 3H).

Example 3 Allyl 6 (E) - (2-pyridyl) methylene penicillanate

A mixture of 2.64 g (6.8 mmol) of 2-picolyltriphenylphosphonium chloride and 0.265 g (6.8 mmol) of sodium amide in 6 mL of dry tetrahydrofuran (THF) was stirred for 30 minutes at room temperature. The resulting brown suspension was cooled to -78 ⁰ C, a solution of 1.8 g (7.0 mmol) of allyl 6-oxopenicillanat in 4 ml of dry THF was added in one portion and the mixture stirred at -78 ° C for 3 min touched. The reaction was quenched by addition of saturated sodium chloride solution, extracted with ethyl acetate and the combined organic layers were washed with water (3x), dried (over MgSO 4) and concentrated in vacuo to give 3.3 g crude oil. The oil was purified by chromatography on a silica gel column to give 1.35 g (60.7%) of the desired product as a yellow oil. H-NMR _(CDCl3)

2a 1.85- S-i £ 4u2

24 /

- j O - "

ppm (S): 1.50 (s, 3H), 1.58 (s, 3H), 4.57 (s, 1H), 4.65 (d, 2H), 5.15-6.15 fm, 3H), 6.17 (d, 1H, J = IHz), 6.87 (d, 1H, J = IHz), 7.2-7.4 (m, 2H), 7.60 (t of d, 1H) , 8.62 (d of d); ¹³ C-NMR (CDCl ₃₎ ppm {£): 26.04, 32.99, 62.77, 65.75, 70.01, 70.54, 119.10, 123.24, 124.02, 125 , 86, 131.06, 136.34, 144.66, 149.94, 152.13, 167.54, 168.73.

2a 1.8b-

Example 4

The application of the procedure of Example 3, but with the appropriate Wittig

3 θ reagent of the formula (CgH ₅ J ₃ PCH ₂ R Cl instead of 2-picolyl-triphenylphosphonium

chloride, provides the following compounds.

^ COOCH ₂ CH = CH ₂

R "

Elution% time, min Temp. ⁰ C Isgsrii

Cl

30

-78 to +25

13 (E) +3.2 (Z) H-NMR _(CDCl3) ppm (delta);

E-isomer ; 1.5 (s, 3H), 1.6 (s, 3H), 4.5 (s, IH), 4.65 (d, 2H), 5.1-6.2 (m, 311), 5.73 (d, IH), 6.82 (d, IH) , Z isomer ; 1.5 (s, 3H), 1.6 (s, 3H), 4.57 (s, IH), 4.7 (d, 2H), 5.15-6.24 (m, 3H), 5.77 (s, IH), 6.37 (s, IH) ,

Example 4 (continued)

Time, min Temp. ⁰ C

CH ₃ S

-78

(A)

10

25

Elutionslsgsm. *

% 4.

Yield ⁴ " ¹ H-NMr (CDClI ppm (delta);

16 (E) 2 (Z) 15 (E + Z)

32 (E)

Performed with formylmethylenetriphenylphosphorane in benzene.

Z isomer ; 1.48 (s, 311), 1.60

(s, 311), 2.58 (s, 3H), 4.42

(s, IH), 4.60 (d, 2H), 5.0-6.0

(m, 3H), 5.58 (s, IH), 6.42 (s, IH).

E-isomer : 1.45 (s, 3H), 1.60

(s, 3H), 2.40 (s, 3H), 4.60

(s, IH), 4.80 (d, 2H), 5.0-6.1

(m, 3H), 5.72 (d, IH), 6.90 (d, IH).

1.50 (s, 3H), 1.60 (s, 3H), 4.52

(s, IH), 4.62 (d, 2H), 5.1-6.0

(m, 3H), 5.90 (d, IH), 6.80 (d, IH), 9.73 (d, IH).

Example 4 (continued)

YY η

Elution R ³

Time, min Tenp. ° C lscrsm. * Yield ¹ H-NMR (CDCl ₃ ) ppm (delta):

2-quinolyl 3 -78 B 80 (Z) 1.5 (s, 3H), 1.62 (s _f 3H), 4.6

(s, IH), 4.7 (d, 2H), 5.1-6.2 (m, 3H), 6.35 (d, IH), 7.0 (d, IH) , 7.2-8.2 (m, 611).

C ₅ H ₅ S 1 -78 B 12 (E, Z) 1.5 (s, 2.2h), 1:55 (s, 0.8H),

1.65 (s, 2.2H), 1.7 (s, 0.8H), 4.5 (s, IH), 4.5-4.8 (m, 2H), 5.1-6.1 (m, 4H), 6.8 (s, 0.2611), 7.15 ( d, 0.74H), 7.4 (m, 5H).

A ^ 3 -78 B 20 (E) 1.45 (s, 3H), 1.55 (s, 3H), 4.55

Cl (s, IH), 4.65 (d, 2H), 5.2-6.1

(m, 3H), 6.15 (d, IH), 6.75 (d, III), 7.05-7.75 (m, 3H).

, R ³ Time, min TENP. ⁰ C Elution lsgsm. * ^% Yield ^C 6 ^H 5 5 -78 A 4 (Z)

Example 4 (continued)

¹ H-NMR (CDCl ₃ ) ppm (delta);

1.5 (s, 3H), 1.6 (s, 3H), 4.6 (s, IH), 4.68 (d, 2H), 5.1-6.1 (m, 3H), 5.77 (s, IH), 6.57 (s, IH) , 7.2-7.64 (ro, 3H), 7.64-8.16 (m, 2H)

 30 (E) 1.52 (s, 3H), 1.62 (s, 3H), 4.60,

(s, IH), 4.69 (d, 2H), 5.1-6.3 ^

, (m, 3H), 6.1 (d, IH), 7.03 (d,

IH), 7.4 (s, 5H). CH, CH,

^J 3-78 C 14 (E) 1.55 (s, 3H), 1.65 (s, 3H), 2.55

(s, 6H), 4.65 (s, IH), 4.75 (d, 2H), 5.1-6.1 (m, 3H), 6.25 (d, IH), 6.9 (s, IH), 7.0 (d, lH). 10 (E, Z)

Example 4 (continued)

Elution time, min Temp. ⁰ C lsgsm. *

Yield"

H-NMR (CDCl ₃ ) ppm (delta):

-78

-78

OCH ₀ CH = CH-,

^Δ 5 -78 A ¹

(E)

(E)

27

1.5 (s, 311), 1.6 (s, 311), 3.85 (s, 3H), 4.55 (s, IH), 4.7 (d, 2H), 5.1-6.1 (m, 3H), 6.2 (d, IH) , 7.0-7.5 (m, 3H), 8.2 (t, IH).

1.5 (s, 3H), 1.6 (s, 3H), 4.4-4.8 (m, 5H), 5.1-6.3 (m, 6H), 6.2 (d, IH), 7.15 (d, 2H), 7.4 (d, IH), 8.15 (t, IH).

E-tsomer ; 1.5 (s, 3H), 1.6 (s, 3H), 2.6 (s, 3H), 4.6 (s, IH), 4.65 (m, 2H), 5.1-6.2 (m, 3H), 6.2 (d, IH) , 6.85 (d, IH), 7.0-7.7 (m, 3H).

Example 4 (continued)

R "

Time, min Temp. ⁰ C

ft

OCH.

-78

-78

* A = hexane / ethyl acetate (9: 1), A ₁ = hexane / ethyl acetate (7: 3), B = chloroform,

C = chloroform / ethyl acetate (9: 1), D = chloroform / ethyl acetate (99: 1).

Ε isomer (R is anti to β-lactam) Z isomer (R is syn to β-lactam) Elutionslsqsm. *

Repeat ^ 1 H-NMR (CDCl ₃ ) ppm (delta) :

50

13

Ε isomer : 1.5 (s, 3H), 1.6 (s, 3H), 4.55 (s, IH), 4.67 (m, 2H), 5.0-6.2 (m, 3H), 6.15 (d, IH), 6.95 ( d, IH), 8.4-8.8 (m, 3H) \

E isomer : 1.5 (s, 3H), 1.7 (s, 3H), 4.55 (s, IH), 4.7 (m, 2H), 5.1-6.2 (m, 3H), 6.2 (s, IH), 6.5- 7.0 (m, 3H), 8.5 (d, IH).

24/6 //

Example 5 Sodium 6 (E) -fc-pyridyl) -methylenepicillanate

A mixture of 0.120 g (0.38 mmol) of allyl 6- (E) - (2-pyridyl) methylene penicillanate, 20 mg of tetrakis (triphenylphosphine) palladium (0) and 20 mg of triphenylphosphine was dissolved in 3 ml of ethyl acetate, and To this was added 0.76 ml (0.38 mmol) of 0.5 M sodium 2-ethylhexanoate in ethyl acetate under a nitrogen atmosphere. The mixture was stirred at room temperature for 2 h, the precipitate was collected by filtration, washed with ethyl acetate and dried in vacuo to give 57 mg (48%) of the title compound as a yellow solid. ¹ H NMR (D ₂ O) ppm (i): 1/55 (s, 6H), 4.33 (s, 1H), 6.17 (d, 1H, J = O, 5 Hz), 7, 03 (d, 1H, J = O, 5Hz), 7.17-8.07 (m, 3H), 8.57 (m, 1H); IR spectrum (KBr): 3433, 1756, 1605 cm " ¹ .

Example 6

Use of the appropriate starting material selected from the allyl esters provided in Example 4, in the procedure of Example 5, provides the following sodium salts as well.

R ' Cl

isomer

(E)

yield

95

Cl

CH ₃ S

(Z)

(E)

89

80

Physical properties

yellow solid, H-NMR (D_O) ppm (delta): 1.50 (s, 3H), 1.58 (s, 3H), 4.3 (s, IH), 5.83 (d, IH), 7.1 (d, IH) ; Infrared Spectrum (KBr) cm " ¹ : 1573, 1607, 1688, 1775, 3460.

Infrared Spectrum (KBr) cm " ¹ : 1580, 1609, 1679, 1753, 3491.

white solid 7H-NMR (D ₂ O) ppm (delta): 1.48 (s, 3H), 1.56 (s, 3H), 2.50 (s, 3H), 4.20 (s, IH), 5.88 (s, IH), 7.2 (s, IH); Infrared Spectrum (KBr) cm " ¹ : 1396, 1606, 1749, 2926, 2963, 3552.

ρ 1 jn » / A l.oU"

Example 6 (continued)

^C 6 ^H 5 ^C 6 ^H 5

Isomer (Z) (E)

Example 6A

yield

60

80

Physical properties light yellow powder

white powder, ¹ H NMR (D ₂ O / DMSO) ppm (delta): 1.5 (s, 6H) / 4.25 (s, IH), 6.1 (d, IH), 7.0 (d, IH), 7.4 (s , 5H); Infrared Spectrum

(KBr) cm " ¹ : 1626, 1642, 1655, 1742, 3434.

6-phenylthiomethylene-penicillanic

A mixture of 93 mg (0.26 mmol) of allyl-6-phenylthiomethylenepicillanate (mixture of isomers) and 10 mg each of tetrakis (triphenylphosphine) palladium (0) and triphenylphosphine was dissolved in 1 ml of ethyl acetate and 0.52 ml of 0.5 m Sodium 2-ethylhexanoate in ethyl acetate was added at room temperature and the resulting mixture was stirred under nitrogen for 10 h. Since very little salt precipitated, the mixture was quenched with water and extracted with methylene chloride. The aqueous layer was acidified (pH 3.5) and extracted with methylene chloride. The dried extracts were concentrated in vacuo to afford 63 mg (75%) of the free acid as an isomer mixture. H-NMR _(CDCl3) ppm (£): 1.5 (s, 2.1H), 1.55 (s, 0.9H), 1.6 (s, 2.1H), 1.65 (s , 0.9H), A ₁ A

(s, 0.7H), 4.5 (s, 0.3H), 5.38 (d, 0.7H), 5.7 (s, 0.3H), 6.7 (s, 0.3H ), 7.1 (d, 0.7H), 7.5 (m, 5H).

Example 7 Allyl-1,1-dioxo-6 (E) - (2-pyridyl) -methylenepicillanate

To a solution of 1.30 g (4.09 mmol) of allyl-6 (E) - (2-pyridyl) methylene penicillanate in 15 mL of methylene chloride was added 1.70 g (8.2 mmol) of 80-85% pure m.sup. Chlorperbenzoic acid was added and the mixture stirred under nitrogen for 3 h at room temperature. After quenching with saturated sodium thiosulfate solution and water, the mixture was extracted with methylene chloride, the organic layer adjusted to pH 7.5 with saturated sodium bicarbonate solution, washed with water, dried (over MgSO 4) and the solvent evaporated in vacuo to 1.4 g (98%) to give product as a yellow oil. The oil was purified by silica gel column chromatography eluting with 7: 3 hexane / ethyl acetate to give 0.78 g (55%) of the title sulfone as colorless crystals. ¹ H-NMR _(CDCl3) ppm (s): 1.48 (s, 3H), 1.63 (s, 3H), 4.45 (s, 1H), 4.73 (d, 2H), 5 , 1-6,2 (m, 3H), 5,77 (d, 1H, J = O, 5Hz), 7,27 (d, 1H, J = O, 5Hz), 7,1-8,1 ( m, 3H), 8.6 (m, 1H); ¹³ C-NMR _(CDCl3) ppm (S): 18,53, 20,43,63,18, 64,25,66,63, 72,04, 119.91, 124.64, 126, O3, 130 , 68, 132, 83, 136, 77, 150, 31, 166, 86, 168, 11. Infrared (KBr) cm " ¹ : 1323, 1586, 1759, 1783, 3437.

Example 8 Allyl-1, 1-dioxo-6 (E) - (2-hydroxyethylidene) penicillanate

To a solution of 0.190 g (0.61 mmol) of allyl-1,1-dioxo-6- (E) -formylmethylenepenicillanate in 4 mL of dry tetrahydrofuran at -78 ° C was added 0.61 mL (0.61 mmol) 1 m Diisobutylaluminum hydride in hexane. The mixture was stirred at -78 ⁰ C for 10 min, quenched with methanol at room temperature for 20 min and filtered. The filtrate was concentrated in vacuo to give 0.258 g of crude product which was mixed with water.

was extracted with chloroform and the organic layer was dried (over MgSO 4). Evaporation of the chloroform provided 160 mg of material which was further purified by silica gel column chromatography eluting with 4: 1 chloroform / ethyl acetate to afford 113 mg (60%) of the title compound. H-NMR (CDC ^) ppm (£): 1.40 (s, 3H), 1.60 (s, 3H), 2.60 (bs, 1H), 4.3 (m, 2H), 4, 4 (s, 1H), 4.7 (d, 2H), 5.1-6.0 (m, 3H), 5.25 (d, 1H), 6.38 (m, 1H).

Example 9

Starting from the appropriate 6-methylene penicillanate ester (n = O) of Example 4 as starting materials in the procedure of Example 7, the following compounds are obtained.

Isomer% yield

Silica gel eluting

Physical Properties ¹ H-NMR _(CDCl3) ppm (delta);

CHO

(E)

2-quinolyl *

(E)

  1.42 (s, 3H), 1.60 (s, 3H), 4.50

(s, IH), 4.70 (d, 2H), 5.1-6.0

(m, 3H), 5.32 (s, IH), 7.1 (m,

IH), 9.73 (d, IH).

CHCl ₃ 1.5 (s, 3H), 1.6 (s, 3H), 4.45

(s, IH), 4.7 (d, 2H), 5.1-6.1

(m, 3H), 5.8 (d, IH), 7.3-8.5

(m, 7H).

In a subsequent approach, a 29% yield of sulfon (n = 2) was obtained by elution from a silica gel column containing chloroform containing 2% ethyl acetate and a more polar fraction identified as the sulfoxide (n = 1) in 27% yield.

Example 9 (continued)

R "

6-Chloro-2-pyridyl

CH

CH-

IKN

isomer

(E)

t yield

34

(E)

(E)

8 (79%

raw)

Silica gel eluting

CIICl.

CHCl.

Hexane / ethyl acetate (1: 1)

Physical properties ¹ H-NMR (CDClO) ppm (delta):

1.5 (s, 3H), 1.6 (s, 3H), 4.45 (s _f IH), 4.7 (d, 2H), 5.1-6.3 (m, 3H), 5.75 (d, IH), 7.1-7.8 (m, 4H).

1.5 (s, 3H), 1.6 (s, 3H), 2.55 (s, 6H), 4.5 (s, IH), 4.7 (d, 2H), 5.0-5.9 (m, 3H), 5.7 (d, IH) , 6.9 (s, IH), 7.3 (d, IH).

1.4 (s, 3H), 1.6 (s, 3H), 3.8 (s, 3H), 4.4 (s, IH), 4.65 (d, 2H), 5.3-6.2 (m, 3H), 5.8 (d, IH) , 7.4 (s, IH), 7.8 (d, IH), 8.35 (t, IH).

Example 9 (continued)

OCH ₂ CH = CH ₂

CH.

isomer

(E)

(E)

(E)

% Yield

54

53

40

Silica gel eluting

Hexane / ethyl acetate (1: 1)

CHCl.

CH 2 Cl 2 / ethyl acetate (9: 1)

Physical properties 1 H-NMR (CDCl ₃ ) ppm (delta):

1.45 (s, 3H), 1.6 (s, 3H), 4.4-4.8 (ro, 5H), 5.1-6.3 (m, 6H) 5.65 (d, IH), 7.15 (d, 2H), (d, IH) , 8.15 (t, IH).

1.5 (s, 3H), 1.64 (s, 3H), 2.64 (s, 3H), 4.48 (s, IH), 4.74 (d of t, 2H), 5.3-5.5 (in, 2H), (d, IH ), 7.1-7.2 (m, 2H), 7.25 (d, IH), 7.6 (t, IH). Infrared Spectrum (KBr) cm 1588, 1688, 1728, 1756, 1795, 2931, 2976, 3433.

1.56 (s, 3H), 1.7 (s, 3H), 4.6 (s, IH), 4.85 (m, 2H), 5.2-6.2 (m, 3H), 5.8 (d, IH), 7.4 (d, IH) , 8.5-9.0 (m, 3H).

-1

Example 9 (continued)

Silica gel Physical properties

³ isomer% yield Eluent ¹ H-NMR (CDCl,) p pm (delta):

OCH

ty

no purification 1.5 ( _s , 3H), 1.7 (s, 3H), 3.9

required, _. , _. . ". . ",, -

(s, 3H), 4.5 fs, IH), 4.75 (m,

2H), 5.2-6.3 (m, 3H), 5.7 (d, IH), 6.7-7.0 (m, 2H), 7.2 (d, IH), 8.5 (d, IH).

247

Example 9A

Allyl-6 (E) - / "(1-oxoquinolin-2-yl) methylene / -1, 1- dioxopenicilanate

Allyl-6 (E) - / (quinolin-2-yl) methylene / -1-oxopenicillanate (as a by-product from the preparation of the corresponding sulfone, see previous example, asterisk) (124 mg, 0.313 mmol) was dissolved in 5 mL of methylene chloride and 195 mg (0.904 mmol) of 80% m-chloroperbenzoic acid were added. The mixture was stirred at room temperature for 48 h, washed with water.

Quenched and extracted with methylene chloride. The

Extracts were washed with saturated sodium bicarbonate solution, water, dried and concentrated in vacuo to a yellow oil. The oil was purified by silica gel column chromatography to give 45 mg (35%) of the titled N-oxide as a yellow solid. H-NMR _(CDCl3) ppm (<f): 1.45 (s, 3H), 1.6 (s, 3H), 4.45 (s, 1H), 4.7 (m, 2H), 5 , 0-6.0

(m, 3H), 5.85 (d, 1H), 7.3-8.0 (m, 7H).

Example 10 Sodium 1, 1-dioxo-6 (E) - (2-pyridyl) -methylenepicillanate

A mixture of allyl-1, 1-dioxo-6 (E) - (2-pyridyl) -methylenepicillanate (0.14 g, 0.4 mmol), 20 mg tetrakis (triphenylphosphine) palladium (O) and 20 mg triphenylphosphine were dissolved in 2 ml of ethyl acetate and under nitrogen, 0.8 ml (0.4 mmol) of a 0.5M solution of sodium 2-ethyl-hexanoate in ethyl acetate was added. The resulting mixture was stirred at room temperature for 5 minutes. The resulting precipitate was filtered, washed with ethyl acetate and dried to give 0.13 g (95%) of the sodium salt as a yellow solid. ¹ H-NMR (D ₂ O) pmm (delta): 1.50 (s, 3H), 1.60 (s, 3H), 4.23 (s, 1H), 5.90 (d, 1H, J = IHz), 7.1-8.0 (m, 4H), 8.57 (m, 1H). IR spectrum (KBr) cm " ¹ : 1590, 1621, 1770, 3454.

24? 677

Example 11 »

Allyl-1,1-dioxo-6 (E) - (I-oxo-2-pyridyl) -methylenepicillanate

A solution of allyl 1,1-dioxo-6 (E) - (2-pyridyl) methylene penicillanate (100 mg, 0.286 mmol) in 5 mL of methylene chloride was treated with m-chloroperbenzoic acid (120 mg, 0.59 mmol) and added Room temperature stirred for 3 days. The mixture was quenched with saturated sodium thiosulfate solution and extracted with ethylene chloride. The organic layer was neutralized with saturated sodium bicarbonate solution, washed with water, dried and concentrated to give 82 mg of yellow oil. The yellow oil was purified by silica gel column chromatography using ethyl acetate as the eluent to give 22 mg (21%) of the title compound and 14 mg (13%) of a by-product, 2,3-epoxypropanyl-1,1-dioxo-6 (E. ) - (1-oxo-2-pyridyl) methylene penicillanate. Allyl-1,1-dioxo-6 (E) - (1-oxo-2-pyridyl) methylene-penicillanate: ¹ H-NMR _(CDCl3) ppm (S): 1.5 (s, 3H), 1, 6 (s, 3H), 4.45 (d, 1H), 4.7 (d, 2H), 5.1-6.0 (m, 3H), 5.8 (s, 1H), 7.1 -8.4 (m, 5H).

example

The compounds of the following formula were also obtained by hydrolysis of the appropriate allyl ester by the method of Example 10.

R ³ CH

O "* '' 'COON a

The use of potassium 2-ethylhexanoate instead of sodium 2-ethyl-hexanoate J: provided the corresponding potassium salts of the above formula

R '

HIGH,

isomer

(E)

Issue 53

Physical Properties

¹ H NMR (D ₂ O) ppm (delta): 1.5 (s, 3H), 1.6 (s, 3H), 4.24 (s, IH), 4.30 (dd, 2H), 5.75 (s, IH), 7.20 (d, IH);

Infrared spectrum. (KBr) cm ": 1618, 1674, 1767, 3408, 3440.

Example 12 (continued)

R "

2-quinolyl

isomer

(E)

% Yield

6-chloro-2-pyridyl

(E)

Physical Properties

¹ H-NMR (250 MHz, DMSO) ppm (delta): 1.45 (s, 3H), 1.48 (s, 3H), 3.82 '(s, IH), 5.96 (s, IH), 7.55-8.55 (m, 7H);

Infrared Thermoelectricity (KBr) cm ": 1619, 1771, 3437.

¹ H-NMR (250 MHz, DMSO) ppm (delta):. 1.56 (s, 3H), 1.64 (s, 3H), 4.34 (s, IH), 6.18 (s, IH), 7.45-8.0 (m, 4H);

, Infrared spectrum (KBr) cm ": 1624, 1771, 3433.

Example 12 (continued)

CH ₃ CH ₃

OCH

Ι-ΟίΧθ-2-quinolyl

isomer

(E)

(E)

% Yield

75

55

(E)

56

Physcal properties

¹ H-NMR (250 MHz, D ₂ O) ppm (delta): 1.52 (s, 3H), 1.60 (s, 3H), 2.5 (s, 6H), 4.32 (s, IH), 6.16 (d, IH ), 7.35 (d, IH), 7.65-7.75 (m, IH);

Infrared spectrum (KBr) cm ": 1629, 1781, 3403.

¹ H-NMR (250 MHz, _CDCl3) ι ppm (delta) 1:42 (s, 3H), 1:48 (s, 3H), 3.85 (s, IH), 3.95 (s, 3H), 5.85 (d, IH) , 7.45 (d, IH), 7.45-8.0 (m, 2H), 8.22 (m, IH);

Infrared spectrum (KBr) cm ": 1617, 1767, 3454.

Infrared spectrum (KBr) cm ~:. 1553, 1622, 1769, 3413.

Example 12 (continued)

CX

(Kaliuitisalz) hygroscopic

, ₀ CH ₀ CH = CH,

isomer

(E + Z)

% Yield

99

Physical Properties

¹ H-NMR (DMSO) ppm (delta): 1.35 (s, 6H), 1.4 (s, 1.4H), 1.44 (s, 6H), 1.48 (s, 1.4H), _v 3.7 (s, IH), 4.55 (d, IH), 4.75 (d, IH), 5.15-5.5 (m, 2H), 5.67 (s, 0.6H), 5.8 (s, 0.4H), 5.8-6.0 (m, 0.5H), 6.0 -6.2 (m, 0.5H), 6.75-7.0 (m, 2H), 7.4-7.6 (m, IH), 8.2 (d, 0.5H). Infrared spectrum * (KBr) cm: 1613, 1760, 3429.

Example 12 (continued)

OCH.

isomer

(E)

% Yield

76 Physical Properties

H-HMR (D ₂ O) ppm (delta): 1.8 (s, 3H), 1.9 (s _f 3H), 4.56 (s, III, 7.2 (IH), 7.34 (IH), 7.62 (IH), 8.68 ( IH).

¹³ C-NMR (D ₂ O) ppm (delta): 20.9, 22.7, 58.7, 64.6, 68.7, 74.6, 113.9, 115.9, 134.0, 134.2, 154.6, 155.1, 169.4, 173.7 175.6.

Example 13 Pivaloyloxymethyl-6 (E) - (methylthio) methylene penicillanate

A mixture of 2.4 mmol of (methylthiomethyl) triphenylphosphonium chloride, 2.4 mmol of sodium amide in 5 ml of dry tetrahydrofuran (THF) was stirred at room temperature for 20 minutes. To the resulting yellow solution, a solution of 788 mg (2.4 mmol) was added pivaloyloxymethyl 6-oxopenicillanat in 10 ml of dry THF at -78 ⁰ C. The mixture was stirred at -78 ⁰ C 1 min, poured into saturated ammonium chloride solution and extracted with ethyl acetate. The organic layer was dried (over Na 2 SO 4) and the solvent removed in vacuo to afford 774 mg of crude product, which was purified by silica gel column chromatography eluting with chloroform to give 220 mg (24.5%) of pure product , ¹ H-NMR _(CDCl3) ppm (£): 1.25 (s, 9H), 1.50 (s, 3H), 1.65 (s, 3H), 2.45 (s, 3H), 4 , 45 (s, 1H), 6.85 (m, 3H), 7.0 (d, 1H).

Example 14

Pivaloyloxymethyl-6- (E) methylsulfonylmethylenepicillanate 1-oxide (A) and the corresponding 1,1-dioxide (B)

To a solution of 215 mg (0.58 mmol) of pivaloyloxymethyl 6- (E) - (methylthiomethyl) methylene-penicillanate in 5 mL of methylene chloride was added 375 mg (1.74 mmol, 3 equiv.) Of 80% m-chloroperbenzoic acid. The mixture was stirred at room temperature for 4h, quenched with water, saturated sodium thiosulfate solution, sodium bicarbonate and extracted with chloroform. The organic phase was washed three times with water, dried (over MgSO 4) and concentrated in vacuo to give 200 mg of product mixture. The crude mixture was purified by silica gel chromatography eluting with chloroform / ethyl acetate (9: 1) to give 25 mg of the 1-oxide (A).

and 45 mg of the 1,1-dioxide product (B).

(A): ¹ H-NMR _(CDCl3) ppm (S): 1.21 (s, 9H), 1.3 (s, 3H), 1.7 (s, 3H), 3.1 (s, 3H), 4.7 (s, 1H), 5.8 (AB quadruplet, 2H), 5.85 (d, 1H), 7.1 (d, 1H); IR spectrum (CHCl ₃ ) cm ^-1 : 1333, 1759, 1807, 2927, 296Ο.

(B): ¹ H-NMR _(CDCl3) ppm (£): 1.2 (s, 9H), 1.45 (s, 3H), 1.6 (s, 3H), 3.15 (s, 3H), 4.5 (s, 1H), 5.6 (d, 1H), 5.8 (AB quadruplet, 2H), 7.2 (d, 1H): IR spectrum (CHCl ₃ ) cm ^-1 : 1324, 1758, 1800, 2929, 2956.

Example 15

Allyl 6-oo (N-methylpyrrol-2-yl) hydroxymethyl-1, 1-dioxo penicillanate

Allyl-6-oi-bromo-1,1-dioxopenicillanate (520 mg, 1.48 mmol) was dissolved in 10 mL of dry tetrahydrofuran (THF) and cooled to -78 ° C. A solution of methyl magnesium bromide (0.52 ml, 2.85 M in THF) was added and the mixture is stirred at -78 ⁰ C Gemisch5min. N-methylpyrrole-2-carboxaldehyde (162 mg, 0.16 mmol) was added and stirring continued at -78 ° C for 20 min. The mixture was poured into saturated ammonium chloride solution, extracted with ethyl acetate and the organic layer dried over MgSO ₄ . Evaporation of the solvent in vacuo gave 466 mg of crude product, which was purified by silica gel chromatography eluting with chloroform / ethyl acetate (9: 1) to give 180 mg (32%) of the pure title compound. H NMR (CDCl ₃₎ ppm (i): 1.4 (s, 3H), 1.62 (s, 3H), 3.68 (s, 3H), 4.0-4.4 (m, 1H ), 4.42 (s, 1H), 4.5-4.8 (m, 3H), 5.0-6.0 (m, 4H), 6.0-6.7 (m, 3H).

Example 16

Allyl-6 (E) - (N-methylpyrrol-2-yl) methylene-1,1-dioxo penicillanate

Allyl-6 - - / - (N-methylpyrrol-2-yl) hydroxy-methyl 1-1,1-dioxopenicillanate (180 mg, 0.47 mmol) was dissolved in 3 mL tetrahydrofuran and 0.15 mL acetic anhydride and 0 , 2 ml of pyridine were added. The mixture was stirred for 1 h at room temperature. The reaction was quenched with water and extracted with ethyl acetate. The organic layer was dried and solvent removed in vacuo to give 162 mg of starting material. This was dissolved in methylene chloride (3 ml) and 0.15 ml of acetyl chloride and 0.2 ml of pyridine were added. The mixture was stirred for 2 h at room temperature and worked up as before to give 140 mg of crude product, which was purified by silica gel column chromatography to afford 72 mg (42%) of pure product. Recrystallization from ethyl acetate gave colorless needles. ¹ H-NMR _(CDCl3) ppm (£): 1.45 (s, 3H), 1.65 (s, 3H), 3.7 (s, 3H), 4.4 (s, 3H), 4 , 6-4.9 (m, 2H), 5.1-6.4 (m, 4H), 6.6-7.0 (m, 2H), 7.5 (dd, 1H).

Example 17

Sodium 6 (E) - (N-methylpyrrol-2-yl) methylene-1, 1-dioxo penicillanate

A solution of 46 mg of allyl-6 (E) - (N-methylpyrrol-2-yl) -methylene-1,1-dioxopenicillanate, 5 mg of tetrakis (triphenylphosphine) palladium (0), 4 mg of triphenylphosphine and 1 ml of methylene chloride was stirred under nitrogen for 5 min. The resulting mixture was diluted with 1 ml of ethyl acetate and 0.25 ml of sodium 2-ethyl-hexanoate in ethyl acetate was added. After stirring at room temperature for 1 h, the mixture was filtered and the precipitate was extracted with ethyl acetate and ethyl acetate.

ether to give 30 mg of yellow solid. ¹ H-NMR (D ₂ O) ppm (S.) ι 1/50 (s, 3H), 1.60 (s, 3H), 3.65 (s, 3H), 4.10 (s, 1H) , 5.4 (s, 1H), 6.1-6.5 (m, 1H), 7.0 (s, broad, 2H), 7.2-7.4 (m, 1H); IR spectrum (KBr) cm " ¹ : 1568, 1616, 1660, 1745, 3465.

Example 18

Use of the appropriate aldehyde of formula R CHO in the procedure of Example 15 provides the corresponding compounds of the following formula

OH 0 0

silica gel

• N-

, - Kieseigel- ₁

R% A eluent usbeute * H-NMR _(CDCl3) ppm ():

³⁰ Å "· 6-alpha isomer: 1.4 (s,

0.39H), 1.5 (s _f 2.61H),

1.62 (s, 0.39H), 1.7 (s,

2.61H), 3.0 (bs, IH),

4.0-4.4 (m, IH), 4.45

(s, IH), 4.5-4.9 (m,

2H), 5.1-6.1 (m, 4H),

6.3-6.6 (m, 2H), 7.45 (m, IH).

Il

Mixture of 6 - </, 8R- and

6- </, 8S-isomers

PH ^{38 A} 1.4 (s, 3H), 1.60 (s,

? "3H), 2.50 and 2.60 (s,

^C0CH 3 3H), 4.1-4.4 (m, Ih) -,

4.4 and 4.5 (s, IH), 4.6-5.0 (m, 3H), 5.Ι δ.Ο (m, 4H), 6.0-7.2 (m, 4H).

_ _bb _

Example 18 (continued)

, ο silica gel ₁

R% yield elutions m. * H-NMR _(CDCl3) ppm (delta)

^C 6 ^H 5 ^{78 A} 1.4 (s, 3H) -, 1.6 (s, 3H),

3.28 (bs, IH), 3.8-4.2 (m, IH), 4.35 (s, IH), 4.45-4.8 (m, 3H), 5.0-6.1 (m, 4H), 7.3 (s, 5H).

³³ B 1.3 (s, 3H), 1.55 (s,

3H), 4.04 (bs, IH), 4.35 (s, IH), 4.5-4.85 (m, 3H), 5.1-6.1 (m, 4H), 7.1-7.4 (m, IH), 7.6-8.0 (m, lH), 8.2-8.7 (m, 2H).

³⁵ B 1.3 (s, 3H), 1.55 (s, 3H),

4.0 (m, IH), 4.35 (s, IH), 4.4-6.8 (πι, 3Η), 5.1-6.2 (m, 4H), 7.2-7.5 (m, 2H), 8.2-8.6 (m, 2H).

⁵⁰ B 1.25 (s, 0.75H), 1.35 (s,

2.25H), 1.55 (s, 3H), 3.3 (bs, IH), 4.05 (dd, IH), 4.3 (s, IH), 4.3-4.8 (m, 3H), 5.0-6.2 (m, 4H), 6.8-7.4 (iu, 3H).

- b b -

Example 18 (continued)

Silica gel ..

% Yield eluent * H-NMR _(CDCl3) ppm ():

CH ₂ = CH 21 1.4 (s, 3H) ; 1.6 (s, 3H),

3.0 (bs, IH), 3.7-4.0 (m, IH), 4.4-4.8 (m, 4H), 5.0 ^ 6.3 (m, 6H). ,

25 D 1.36 Cs, 1.5H), 1.40 (s,

1.5H), 1.60 (s, 1.5H), CH ₃ 1.65 (s, 1.5H), 3.7 (s,

3H), 4.0-4.5 (m, 2H), 4.4 (s, IH), 4.5-4.8 (m, 2H), 5.0-6.0 (m, 4H), 6.7 (s, IH), 6.85 (s, IH) ,

N 52 D 6-alpha, 8S isomer: 1.4

(S, 3H), 1.6 (s, 3H), 4.38 (dd, IH), 4.43 (s, IH), 4.67-4.75 (m, 2H), 4.76 (d, IH), 5.3-5.5 (m, 2H ), 5.63 (d, IH), 5.85-6.05 (m, IH), 7.4 (d, IH), 7.8 (d, IH). 6-alpha, 8R isomer: 1.36 (s, 3H), 1.60 (s, 3H), 4.22 (d, IH), 4.4 (s, IH), 4.65 (m, 2H), 4.88 (s, IH), 5.25 -5.5 (m, 2H), 5.55 (d, IH), 5.8-6.0 (m, IH), 7.35 (d, IH), 7.75 (d, IH).

Example 18 (continued)

13 silica gel ₁

R% Yield Eluent * H-NMR (CDCl -,) ppm ();

C ₆ H ₅ -NN 42 E 6-alpha, 8S isomer: 1.44

N \ Cs, 3H), 1.62 (s, 3H),

3.68 (bs, IH), 4.31 fdd, IH), 4.5 Cs, IH), 4.74 (d, 2H), 4.86 (d, IH),

5.4 (m, 3H), 5.9 Cm, IH),

7.5 (m, 3H), 8.02 (s, IH), 8.14 (in, 2H), IR: 3482, 1802 cm ^-1 .

5 7 crude A (less polar polar 6-oCt 8S)

lares iso-9wp _ner) isomer: 1.41 (s, 3H),

ρ F (more polar i. _{6 (s,} 3H), 4.45 (s, IH),

Isomer)

, 4.4-4.8 (m, 4H), 5.2-

5.6 (m, 3H), 5.7-6.3 (m, IH), 7.35 (t, IH), 8.85 (d, 2H)

 'Polareric 6-alpha, 8R-iEsomer: 1.45 (s, 3H),

1.6 (s, 3H), 4.4 (s, IH), 4.45 (dd, IH), 4.7-4.9 (m, 2H), 4.95 (d, IH), 5.2-5.6 (m, 3H), 5.7-6.3 ( in, IH), 7.35 (t, IH), 8.85 (d, IH).

O / /

Example 18 (continued)

%Yield

Silica gel eluent *

25%, I.Frakt. (one isomer) 12%, 2nd fract. (Mixture of 2 isomers)

56

^70:30 Isonerenge

mixed

22

1: 2

isomeric

mixed

¹ H-NMR _(CDCl3) ppm ():

1st fraction: 1.4 Cs, 3H),

1.5 Cs, 3H), 4.2-4.4 (m, 2H), 4.5-5.0 (m, 3H), 5.1-6.1 Cm, 6H), 7.6 Cd, IH), 8.87 Cd, IH), 9.23 (s, IH) ,

1.4 Cs, 3H), 1.57 (s, 3H), 4.25 (m, IH), 4.37 Cs, 0.7H), 4.42 (s, 0.3H), 4.75 (m, 2H), 4.8 (d, 0.3H), 4.85 (d, 0.7H), 5.25-5.5 (m, 3H), 5.9 (m, IH), 8.52 (m, 2H), 8.84 (m, IH).

1.4 (s, IH), 1.46 (s, 2H),

1.6 (s, IH), 1.63 (s, 2H), 4.12 (m, IH), 4.22 (m, IH), 4.41 and 4.46 (s, IH), *. 4.6-4.8 (in, 2H), '4.95 (d, IH), 5.2-5.5 (m, 3H), 5.9 (m, IH), 6.95 and 7.05 (s, IH), 7.28 and 7.36 (s, IH) ,

The starting aldehyde used was 1-diethoxymethylimidazole-2-carboxaldehyde.

Example 18 (continued)

silica gel

_{R 13% yield} eluent * ¹ H-NMR _(CDCl3) ppm (£):

A - chloroform / ethyl acetate (9: 1)

B - ethyl acetate / chloroform (7: 3)

C - chloroform / ethyl acetate (19: 1) D - chloroform / ifethanol (19: 1) E - chloroform

F - chloroform / ethyl acetate (1: 1) G - ethyl acetate

35 G 1.38-1.40 (d, 3H), Iv56-

L ^X · ^{57 {d} ' ^3H) ' 4.20-4.40

(m, 2H), 4.59-4.72 (m, 2H) , 4.86-4.88 (d, 0.5H) 5.04-5.06 (d, 0.5H), 5.26-5.42 (m, 2H) , 5.50-5.62 (m, IH ), 5.82-6.00 (m, IH), 7.50-7.86 (m, IH), 7.90-8.08 (m, IH), 9.02-9.10 (m, IH), infrared spectrum: 1800 cm ".

Example 19

Allyl-6- (furan-2-yl) methylene-1, 1-dioxopenicillanate, (E) and (Z) isomers

To a solution of 310 mg (0.84 mmol) of allyl 6 - <* r- (furan-2-yl) -hydroxymethyl-1,1-dioxopenicillanate in 5 ml of methylene chloride was added 0.14 ml (1 mmol) of triethylamine and 0.1 ml (0.924 mmol) of trifluoromethylsulfonyl chloride and the mixture stirred under nitrogen at room temperature for 2 h. The reaction was quenched with water, extracted with methylene chloride, the extracts dried (over MgSO ₄ ), and solvent evaporated in vacuo to give 330 mg of crude product. Purification by silica gel column chromatography eluting with chloroform provided 130 mg of product which was held after high performance liquid chromatography as a 4: 1 mixture of (E) and (Z) isomers. ¹ H-NMR _(CDCl3) ppm (£): 1.47 (s, 3H), 1.61 (s, 3H), 4.47

(s, 1H), 4.75 (d, 2H), 5.1-6.2 (m, 4H), 5.52 (dd, 1H), 6.8

(m, 1H), 7.15 (d, 1H), 7.6 (d, 1H).

Example 20 Allyl-6- (E) - (N-acetylpyrrol-2-yl) methylene-1,1-dioxopenicillanate

A. Allyl-6- (N-acetylpyrrol-2-yl) acetoxymethyl-1,1-dioxo penicillanate

Allyl-6- (N-acetylpyrrol-2-yl) hydroxymethyl-1,1-dioxopenicillanate (210 mg, 0.51 mmol) was dissolved in 3 ml of tetrahydrofuran and 0.16 ml of acetic anhydride and 0.2 ml of pyridine were added, and the mixture was stirred at room temperature for 24 hours. The reaction was quenched with water, extracted with methylene chloride, the extracts dried and concentrated to give 171 mg (75%) of yellow crystals ¹ H-NMR _(CDCl3) ppm (i): 1.4 (s, 3H) , 1.6 (s, 3H), 2.15 (s, 3H), 2.55 (s, 3H), 4.15-4.3 (dd, 1H), 4.4 (s, 1H), 4.6-4.8 (m, 3H), 5.1-6.0 (m, 3H), 6.1-6.6 (m, 2H), 6.6-7.4 (m, 2H ).

B. The Ν, Ο-diacetate product of Part A, 170 mg (0.38 mmol),

was dissolved in methylene chloride, and 47 mg (0.38 mmol) of 1-S-diazabicyclo / 3: 3-pentyn-5-ene (DBN) was added. The mixture was stirred at room temperature for 1 h. Water was added and the mixture was extracted with methylene chloride. The extracts were dried and concentrated to give 158 mg of oil, which was purified by silica gel chromatography using 2% ethyl acetate in chloroform as eluent to give 108 mg of product as a pale yellow oil. ¹ H-NMR _(CDCl3) ppm (£): 1.5 (s, 3H), 1.6 (s, 3H), 2.55 (s, 3H), 4.4 (s, 1H), 4 , 65 (d, 2H), 5.0-6.0 (m, 4H), 6.3 (t *, 1H), 6.8 (dd, 1H) _e , 7.2 (m, 1H), 8.2 (d, 1H)

Example 21

Allyl 6 (E) -phenylmethylene-1,1-dioxopenicillanate and the corresponding (Z) isomer

A. Use of 1.98 mmol allyl-6-phenylhydroxymethyl

1,1-dioxopenicillanate, 4.2 mmol acetyl chloride and 0.4 ml pyridine in the method of Part A of the previous example gave: 0.7 g (84%) of allyl 6-phenylacetoxymethyl-i, 1-dioxopenicillanate as a light yellow resin , 'H-NMR (CDClο) ppm (£): 1.3 and 1 ₇ 4 (s, 3H), 1.62 (s, 3H), 2.08 and 2.2 (s, 3H), 4.2 (dd, 1H), 4.4 (s, 1H), 4.5 (d, 1H), 4.65 (d, 2H), 6 _# 25 (m, 1H), 7.3 (m, '5H ).

B. To a solution of the product of Part A (0.7 g, 1.66 mmol) in methylene chloride was added 0.25 mL (1.67 mmol).

1, S-diazabicyclo / S ^ .O./ unde.c-S-ene (DBU), and the mixture was stirred at room temperature for 10 minutes. Water and methylene chloride were added, the layers separated and the organic extracts washed with 0.1N hydrochloric acid, brine and water. The extracts were dried over Na 2 SO 4 and solvent was evaporated to give 660 mg of crude product, which was purified by column chromatography.

was purified on 100 g of silica gel eluting with chloroform to afford 68 mg (11%) of the (Z) -isomer. ¹ H-NMR _(CDCl3) ppm (S): 1.45 (s, 3H), 1.60 (s, 3H), 4.45 (s, 1H), 4.68 (d, 2H), 5 , 37 (d, 1H), 5.1-6.05 (m, 3H), 7.35 (d, 1H), 7.45 (s, 1H). Subsequent eluate fractions gave 100 mg (16.7%) of the (E) -isomer of the title compound as a colorless oil which formed crystals on standing. H-NMR _(CDCl3) ppm (S) '. 1.45 (s, 3H), 1.58 (s, 3H), 4.45 (s, 1H), 4.75 (d, 2H) 5.45 (d, 1H), 5.2-6, 2 (m, 3H), 7.36 (d, 1H), 7.45 (s, 5H).

Example 22

The following compounds were prepared from the appropriate hydroxy compound selected from those of Example 18, following the procedure of Example 19.

R ³ CH

O Ο CH

I '3

% Silica gel eluent yield

22 (E) 19 (Z)

ethyl ether

36

Ethyl acetate hexane (1: 1)

H-NI-IR (CDCl ₃ ) ppm (delta):

(E) isomer : 1.5 (s, 3H), 1.7 (s, 3H), 4.5 (s, IH), 4.7 (d, 2H), 5.5 (d, 1H), 5.1-6.2 (m, 3H), 7.3-7.5

(m, 2H), 7.7-8.0 (m, IH), 8.53-8.83 (m, 2H).

(Z) isomer : 1.48 (s, 3H),

1.6 (s, 3H), 4.5 (s, IH),

4.7 (d, 2H), 5.25 (s, IH), 5.1-6.2 (m, 3H), 6.88 (s, IH), 7.2-7.5 (m, IH), 8.43-9.0 (m, 3H).

Isomeric mixture : 1.45 (s, 3H), 1.62 (s, 3H), 4.5 (s, IH), 4.7 (d, 2H), 5.2-6.2 (m, 4H), 6.75 (s, 0.3H), 7.2-7.5 (m, 1.7H), 7.6-7.85 (m, IH), 8.5-8.83 (m, x 2H).

Λ J Λ

Example 22 (continued)

% Silica gel -

R ³ yield of eluent H-NMR (CDCl 3) ppm ( S) :

CHCl ₃ 1:45 (s, 3H), 1.6 ^(s,: -3H),

_{4 # 4 {Sf 3H) # 4> 7 (d} ^ 2H),

5.0-6.2 (m, 3H), 5.25 (d, IH), 7.0-7.65 (m, 4H).

CH ₂ = CH-18 CHCl ₃ 1.45 (s, 3H), 1.65 (s, 3H),

3.7-4.2 (m, IH), 4.45 (s, 0.6H), 4.5 (s, 0.4H), 4.75 (d, 2H), 5.1-6.4 (m, 7H).

example

Starting from the allyl-6-R ³ -methylene-1,1-dioxopenicillanate esters provided in Examples 19 to 22, the following sodium salts are obtained by the method of Example 17. The corresponding potassium salts are obtained when potassium 2-ethylhexanoate is used.

COCH.

(E)

R ³ CH ^ IR Σ 0 0 W / ^CH 3 O ^% yield 1621 1764 3402 -1 T> ^CH 3 NL ^ COONa j spectrum (KBr). isomer 47 , 1678, 2979, 3472 3: 1 (E): (Z)

cm

56

1622, 1728 1764, 3438 cm " ^X H NMR (D ₂ Q) ppm (delta);

(250 MHz spectrum): 1.5 (s, 3H), 1.6 (s, 3H), 4.25 (s, IH), 5.57 (s, 0.25H), 5.9 (s, 0.75H), 6.65 (m, HI) , 7.0 (d, IH), 7.17 (d, 0.25II), 7.36 (s, 0.75II), 7.78 (m, IH).

(60 MHz spectrum): 1.56 (s, 3H), 1.65 (s, 3H), 2.68 (s, 3H), 4.25 (s, IH), 5.88 (s, HI), 6.5, · (t, IH) , 7.0 (d, IH), 7.6-8.0 (m, IH), 8.2 (s, IH).

Example 23 (continued)

isomer

Yield IR spectrum (KBr)

^C 6 ^H 5 (Z) 91 1565 , 1619, 1667 , 1759 2950 cm , 3453 he (E) 72 1623 3462 , 1778, cm he ^ N I (Z) 91 1303 1761 cm , 1623, 3445 6 (E) ₊ (Z) 93 1623 3448 1778 cm

"H-NMR (D ₂ O) ppm (delta):

(60 MHz spectrum): 1.45 (s, 3H), 1.55 (s, 3H), 4.17 (s, IH), 5.5 (s, IH), 7.0 (s / IH), 7.27-7.6 (m, 311), 7.6-8.1 (m, 2H).

1.55 Js, 3H), 1.62 (s, 3H), 4.23 (s, IH), 6.05 (s, IH), 7.4-8.8 (m, 5H).

, 1.55 (s, 3H), 1.65 (s, 3H), 4.3 (s, IH), 5.6 (s, IH), 6.96 (s, IH), 7.1-9.0 (m, 4H).

1.5 (s, 3H), 1.6 (s, 3H), 4.3 (s, 0.3H), 5.55 (s, 0.3H), 6.0 (d, 0.711), 6.95 (s, 0.3H), 7.2 ^ 7.8 (m , 4.7H).

Example 23 (continued)

CH ₂ = CH-,

isomer

(E)

% Yield IR spectrum (KBr)

(E)

(Potassium salt)

1619, 1773, 3425 cm " ¹

1620, 1773, 3430 cm ^{-1 1} H-NMR (D ₂ O) ppm (delta);

1.55 (s, 3H), 1.60 (s, 3H), 4.2 (s, IH), 5.8 (s, IH), 7.0-7.9 (m, 4H).

1.59 (s, 3H), 1.60 (s, 3H), 4.22 (s, 0.6H), 4.3 (s, 0.4H), 5.53 (s, _v 0.4H) _f 5.75 (s, 0.6H), 5.5-6.2 (m, 3H).

1.5 (s, 3H), 1.6 (s, 3H), 2.5 (s, 3H), 4.28 (s, IH), 6.05 (d, IH), 7.2-7.35 (m, 2H), 7.4 (d, IH) , 7.67 (t, IH).

¹³ C-NMR (D ₂ O) ppm (delta):

20,719, 22,436, 26,032, 68,386, 68,533, 74,489, 120,794, 128,334, 132,843, 140,670, 152,549, 163,012, 174,041, 175,657.

Example 23 (continued)

ox

Isomer yield IR spectrum (KBr)

(E) 92 1614, 1770,

3406 cm " ¹

(Potassium salt) ¹ H-NMR (D ₂ O) ppm (delta);

(D ₂ O + DMSO): 1.5 (s, 3H), 1.6 (s, 3H), 4.1 (s, IH), 5.9 (d, IH), 7.5 (d, IH), 8.5-9.0 (m, 3H ).

Example 24

6 (E) -Phenylmethylene penicillanic acid 1,1-dioxide

To a solution of 0.1 g (0.28 mmol) of allyl-6 (E) -phenylmethylene-1,1-dioxopenicillanate, 20 mg of triphenylphosphine and 20 mg of tetrakis (triphenylphosphine) palladium (0) in 3 mL of ethyl acetate were added 0, 57 ml of 0.5 M sodium 2-ethylhexanoate and the mixture was stirred at room temperature for 1 h. After standing for 65 h in the refrigerator, no drop had formed. The mixture was diluted with ethyl acetate and water, the separated aqueous layer was adjusted to pH 1.8 with dilute hydrochloric acid, extracted with fresh ethyl acetate, the extracts dried (over Na ₂ SO ₄ ), and solvent removed in vacuo to give 62 mg ( 69%) to give product as yellow crystals (from acetone). ¹ H-NMR (CD ₃ COCD ₃₎ ppm (<f): 1.55 (s, 3H), 1.65 (s, 3H), 4.43 (s, 1H), 5.93 (d, 1H ), 7.3-7.9 (m, 6H), 8.7 (bs, 1H); IR spectrum (KBr) cm " ¹ : 1327, 1685, 1737, 1772, 2929, 2961, 3108, 3477.

Example 25

Potassium 6 (E) -i-methylimidazol-2-yl) -methylene-1,1-dioxo penicillanate

A. Allyl-6- (1-methylimidazol-2-yl) acetoxymethyl-1,1-dioxo penicillanate

6- (1-Methylimidazol-2-yl) hydroxymethyl-1,1-dioxopenicillanate (472 g, 1.23 mmol) was acetylated by the method of Example 20, Part A, to give 392 mg (75%) of the acetoxy compound to deliver as a mixture of two isomers. H-NMR _(CDCl3) ppm (£): 1.4 (s, 1.5H), 1.5 (s, 1.5H), 1.6 (s, 1.5H), 1.7 (s , 1H), 2.2 (s, 3H), 3.7 (s, 1.5H), 3.75 (s, 1.5H), 4.0-6.0 (m, 8H), 6, 3-6.5 (m, 1H), 6.8 (m, 1H), 7.0 (m, 1H).

B. allyl-6 (E) -1-methylimidazol-2-yl) methylene-1,1-dioxopenicillanate *

The product obtained in part A, above, (392 mg, 0.920 mmol), _f 5 mL of methylene chloride and 0.115 mL of 1, S-diazabicyclo / 3, oo non-5-ene (DBN) was prepared by the method of Example 20, part

B, converted to the title ester in 53% yield. H-NMR _(CDCl3) ppm (S): 1.5 (s, 3H), 1.6 (s, 3H), 3.7 (s, 3H), 4.35 (s, 1H), 4 , 7 (m, 2H), 5.0-6.1 (m, 3H), 5.7 (d, 1H), 6.9

(m, 1H), 7.1 (d, 1H), 7.2 (m, 1H).

C. The allyl ester (163 mg, 0.45 mmol) obtained in Part B was converted to the title potassium salt by the method of Example 17 but using potassium 2-ethylhexanoate in place of the corresponding sodium salt. The product obtained was 143 mg (87% yield) of the single (E) isomer. ¹ H-NMR (DMSO) ppm (S) ζ 1.38 (s, 3H), 1.45 (s, 3H), 3.8 (s, 4H), 5.68 (s, 1H), 7, 15 (s, 1H), 7.35 (m, 2H); ¹³ C-NMR (DMSO) ppm (lbs): 18.5, 20.2, 32.5, 64.4, 66.2, 70.55, 115.2, 124.6, 129.9, 130, 6, 141.2, 167.9, 169.0. IR spectrum (KBr) cm " ¹ : 1614, 1762, 3428.

Example 26

The procedure of Example 10 was repeated with 50 mg (0.12 mmol) of allyl 6- (3-allyloxy-2-pyridyl) methylene-1,1-dioxopenicillanate using 5.2 mg of triphenylphosphine, 5.2 mg of tetrakis ( triphenylphosphine) palladium (0) and 0.24 mmol of potassium 2-ethylhexanoate (2 molar equivalents of starting allyl ester) in 1.5 ml of ethyl acetate and stirring the resulting mixture for 18h. The ethyl acetate was taken with a pipette and the residue was washed twice with 1 ml portions of ethyl acetate to give a dark solid, 43 mg, which was the dipotassium salt of 6- (E) - (3-hydroxy-2-pyridyl ) methylene-1,1-dioxopeniccanoic acid. H-NMR (D ₂ O) ppm {)): 1.56 (s, 3H), 1.62 (s, 3H), 4.27 (s, 1H), 6.00 (d, 1H), 7 , 29 (m, 2H), 7.80 (d, 1H), 8.04 (d, 1H).

Example 27 Allyl-6- (E) -p-pyrimidinyl) methylenpenicillanate

A. 2-hydroxymethylpyrimidine

A slurry of sodium ethylate prepared from 7.44 g (323 mmol) of sodium metal and 300 ml of absolute ethanol was cooled to room temperature, 18 g (163 mmol) of hydroxyacetamidine hydrochloride and 8.06 ml (81 mmol) of 3-dimethylaminoacrolein were added, and Heated to reflux. Dimethylamine was slowly distilled off with periodic addition of ethanol to maintain the volume of solvent. After refluxing for 9 hours, the mixture was then stirred at room temperature for 18 hours. The solvent was removed under reduced pressure and the residue was placed on a silica gel column and eluted with 19: 1 ethyl acetate / methanol to give 4.1 g of product (46.6% yield).

^B * ^ -pyrimidinylmethyltriphenylphosphonium chloride

2-Hydroxymethylpyrimidine, 4.096 g (37.2 mmol), was dissolved in 80 mL of methylene chloride and 8.6 mL (37.2 mmol) of thionyl chloride was added dropwise (exothermic). The mixture was stirred for 15 min, neutralized with saturated sodium bicarbonate solution and extracted with methylene chloride. The extracts were dried and solvent was evaporated to give 3.67 g of 2-chloromethylpyrimidine. ¹ H-NMR (CDCl3) ppm (S) '. 4.82 (s, 2H), 7.3 (t, 1H), 8.9 (d, 2H).

A mixture of 3.565 g (27.7 mmol) of 2-chloromethylpyrimidine in 30 ml of toluene and 7.269 g (27.7 mmol) of triphenylphosphine was heated at reflux for 18 hours. The resulting precipitate was collected by filtration and dried to give 9.06 g (65% over 2 steps).

_ 82 -

C. A mixture of 2.187 g (5.6 mmol) of the Wittig reagent

The above Part B, 218.4 mg (5.32 mmol) of sodium amide and 30 ml of dry tetrahydrofuran (TKF) were stirred at room temperature for 1.5 h. A solution of 1.44 g (5.65 mmol) of allyl 6-oxopenicillanat in 10 ml of dry THF was added in one portion at -78 ⁰ C, the resulting mixture was stirred for 5 min, poured into saturated ammonium chloride solution and extracted with chloroform. The organic phase was washed with saturated ammonium chloride, brine, dried (over MgSO ₄ ) and the solvent removed in vacuo. The resulting oil was purified by column chromatography on silica gel eluting with 9: 1 chloroform / ethyl acetate to afford 560 mg of the pure product. H-NMR (CDCl3) ppm (£): 1.5 (s, 3HJ, 1.6 (s, 3H), 4.6 (s, 1H), 4.7 (m, 2H), 5.1- 6.3 (m, 3H), 6.2 (d _# 1H), 7.0 (d, 1H), 7.0-7.35 (m, 1H), 8.8 (d, 2H).

Example 28 _r

Allyl ester of 1,1-dioxo-6 (E) - (2-pyrimidinyl-methylene penicillanic acid and potassium salt

A. Allyl 6 (E) - (2-pyrimidinyl) methylene penicillanate (560 mg, 1.69 mmol) and 3-chloroperbenzoic acid, 730 mg (3.38 mmol) were dissolved in 10 mL of methylene chloride and stirred under a nitrogen atmosphere h stirred. The resulting mixture was quenched with water, extracted with methylene chloride, the extracts washed with sodium thiosulfate, neutralized with sodium bicarbonate, washed with brine, dried and concentrated in vacuo to 460 mg crude oil. The oil was purified by silica gel column chromatography eluting with 9: 1 chloroform / ethyl acetate to afford 180 mg of the desired allyl ester as a pale yellow solid. ¹ H-NMR _(CDCl3) ppm {£): 1.45 (s, 3H), 1.65 (s, 3H), 4.5 (s, 1H), 4.75 (m, 2H), 5 , 2-6.3 (m, 3H), .5.75 (s, 1H), 7.1-7.5 (m, 2H), 8.9 (d, 2H).

- Ö3 -

B. Reaction of the above allyl ester with triphenylphosphine, tetrakis (triphenylphosphine) palladium (O) and potassium 2-ethylhexanoate by the method of Example 10 gave the desired potassium salt in 8 9% yield as a pale pink solid. ¹ H NMR (D ₂ O) ppm (S): 1.6 (s, 3H), 1.68 (s, 3H), 4.4 (s, 1H), 6.1 (s, 1H), 7.48 (s, 1H), 7.54 (t, 1H), 8.88 (d, 2H); ¹³ C NMR (D ₂ O) ppm ()): 20.9, 22.7, 68.8, 68.9, 74.6, 124.5, 132.5, 139.9, 160.9, 633, 2, 172.5, 175.5. IR spectrum (KBr) cm " ¹ : 1560, 1615, 1771, 3439.

Example 29

A. To 1.0 g of 6- oc -hydroxypenicillanic acid in 25 ml of methylene chloride is added 50 mg of diisopropylcarbodiimide and 0.5 ml

Added 2,2,2-trichloroethanol. The mixture is stirred overnight, the solvent removed by evaporation in vacuo and the crude product purified by column chromatography on silica gel to provide 2,2,2-trichloroethyl-6- o- hydroxypenicillanate.

B. To 1.0 g of 6- o-C-hydroxypenicillanic acid in 50 ml of dioxane is added 0.5 g of p-toluenesulfonic acid and 0.4 g of dihydropyran

and the mixture warmed to 50 ° C, then stirred at room temperature overnight. Evaporation, solvent and purification by silica gel column chromatography yields 2-tetrahydropyranyl-6-txi-hydroxypenicillanate.

Example 30 Benzyl 6-oxopenicillanate

To a solution of 1.8 ml (0.025 mol) of dimethyl sulfoxide in 2 ml of methylene chloride at -60 ° C was added dropwise a solution of 2.12 ml (0.015 mol) of trifluoroacetic anhydride in 5 ml of methylene chloride. The mixture was stirred at -60 ° C for 20 minutes, a solution of 350 mg (1.14 mmol) of benzyl 6-eC-hydroxypenicillanate in 5 ml of methylene chloride added and further 60 min

stirred at -6O ⁰ C. Tri-ethylamine (0.50 ml) was added, the cooling bath removed, the mixture warmed to ⁰ ° C, poured into ice-water and extracted with methylene chloride. The organic layer was dried, concentrated in vacuo to a small volume which was diluted with benzene and washed three times with ice-water. The benzene layer was dried, solvent removed in vacuo to afford 230 mg (67%) of the product as yellow crystals. The proton NMR spectrum in CDCl ₃ showed the product to be very pure.

Example 31 Benzyl-6- (2-pyridyl) hydroxymethylpenicillanat

A. Benzyl 6-bromo-6 (2-pyridyl) hydroxymethyl penicillanate

A solution of 9.0 g (0.02 mol) of benzyl 6,6-dibrompenicillanat in 200 ml of freshly distilled toluene is cooled to -78 ⁰ C, and 9 ml of 2.2 m t-butyllithium in pentane were added dropwise. The resulting mixture was stirred for 30 minutes, 2.14 g (0.02 mole) of 2-pyridinecarboxaldehyde was added and stirred for a further 40 minutes. The reaction was quenched by the dropwise addition of acetic acid in toluene. After stirring for 1 h, the cooling bath was removed, the mixture warmed to -10 ° C, diluted with 200 mL of toluene, washed with water (5x) and dried (over Na ₂ SO ₄ ). The toluene solution was placed on a Florisil (1 kg) column and eluted with 2: 1 toluene / ethyl acetate. The product fractions were combined and concentrated in vacuo to a brown syrup, 4.2 g, which was used in the next step.

B. The brown syrup from Part A (4.2 g) was dissolved in 50 ml of benzene and 2.65 g of tributyltin hydride were added.

The mixture was heated at reflux for 2 h, additional tributyltin hydride (1.65 g) was added and heating to reflux continued overnight. The solvent was

stripped in vacuo / the residue was washed with hexane and placed on a column containing 5OO g of silica gel and eluted with 2: 1 toluene / ethyl acetate to obtain 425 mg of the title compound. H-NMR _(CDCl3) ppm (S "): 1.35 (s, 3H), 1.7 (s, 3H), 4.0 (dd, 1H), 4.5 (s, IH), 5 , 1 (s, 2H), 5.2 (d, 1H), 5.4 (d, 1H), 7.0-7.8 (m, 3H), 8.5 (m, 1H).

Example 32

6- (2-pyridyl) hydroxymethylpenicillansäure 1,1-dioxide

A. Benzyl 6- (2-pyridyl) hydroxymethyl-1,1-dioxo-penicillanate

To a solution of 0.40 g of benzyl 6- (2-pyridyl) -hydroxymethylpenicillanate in 5 mL of methylene chloride was added 0.20 g of m-chloroperbenzoic acid, and the mixture was stirred for one hour at room temperature. Thin layer chromatography indicated that the mixture contained some sulfoxide. An additional 0.2 g of m-chloroperbenzoic acid was added and the mixture was stirred overnight. The mixture was diluted with methylene chloride, washed again with saturated sodium thiosulfate solution, water, saturated sodium bicarbonate solution, and the organic layer was concentrated in vacuo. The residue was taken up in ethyl acetate, washed with sodium bicarbonate solution, water, brine and dried (over Na ₃ SO ₄ ). Evaporation of the solvent gave 330 mg of the desired benzyl ester as a brown oil, which was purified by silica gel column chromatography eluting with 11: 9 ethyl acetate / hexane to provide 60 mg of yellow oil. ¹ H-NMR _(CDCl3) ppm (S): 1.25 (s, 3H), 1.52 (s, 3H), 4.1 (dd, 1H), 4.5 (s, 1H), 4 , 72 (d, 1H), 5.5 (d, 2H), 5.8 (d, 1H), 7.1-8.0 (m, 3H), 8.5 (m, 1H).

B. A suspension of 118 mg of 10% Pd / C catalyst in 10 ml of tetrahydrofuran (THF) and 4 ml of water was added at 20 min

3 bar hydrogen pressure prehydrated. To this was added 130 mg of the benzyl ester obtained in part A above in 4 ml of the same

Given THF / water mixture. This was hydrogenated at 3.5 bar (50 psi) for 30 minutes. An additional 129 mg of 10% Pd / C was added and hydrogenation continued at 50 psi for 2 h. The catalyst was filtered off, the solvent removed in vacuo and the residue partitioned between water and ethyl acetate. The aqueous layer was freeze-dried to give 85 mg of the desired acid. H-NMR (DO) ppm (S): 1.3 (s, 3H), 1.5 (s, 3H), 4.4 (s, 1H), 5.0-5.35 (m, 2H) , 5.9 (d, 1H); IR spectrum (KBr) cm " ¹ : 1620, 1731, 3407.

C. When the above procedure is used but using a total of only 175 mg of m-chloroperbenzoic acid (an equimolar amount) in Part A, the isolated product is a mixture of the corresponding o £ and β sulfoxides.

Example 33

The following materials are mixed to obtain a powder of uniform composition in the parts by weight given below:

(a) Potassium (6-ei, 8R) -6- (thiazol-2-yl) -acetoxymethyl-1,1-dioxopenicillanate 1, 0

(b) ampicillin trihydrate 1.0

(c) lactose 0.5

(d) polyethylene glycol, avg. MG 4000 3.0

Mixture (1375 mg) is filled into hard gelatin capsules of suitable size to obtain 250 mg capsules of each active ingredient. Higher or lower strength capsules are made by appropriate adjustment of capsule size and fill weight. The relative weights of the active ingredients are adjusted to obtain capsules. in which the weight ratio of active ingredients is other than 1, e.g. the ingredients are in a weight ratio of 0.75, 1.5, 0.5

or 3.0, at a total weight of 1700 mg / capsule, to give 225 mg capsules of (a) and 450 mgs of (b).

Likewise, the other beta-lactamase inhibitors of the invention are formulated with other conventional beta-lactam antibiotics for oral use.

Alternatively, components (a) and (b) in the above formulation are replaced with two parts by weight of 6- / D- (2-amino-2-phenylacetamido-2-pyridyl) -anilino-methyl-o- (2-pyridyl) methylene-1, 1-dioxopenicillanate hydrochloride.

Example 34 Injectable preparation

Equal parts by weight Cefoperazone sodium and potassium 1,1-dioxo-6- (E) - (2-pyrazinyl) methylene penicillanate are combined with 20 parts by weight water. Using standard methods in the pharmaceutical field, the solution is sterile filtered, filled into ampoules, the ampoules are closed loosely with a rubber stopper and the ampoules are freeze-dried on trays. The fill volume is such that each freeze-dried ampule, now sealed under vacuum, contains 500 mg of each active ingredient. Before injection, each vial is filled by injecting 10 ml of sterile water for injection through the rubber stopper and shaking to dissolve. 1 to 10 ml of the solution to be injected are withdrawn through the rubber stopper using a hypodermic needle.

Example 35 , ·

Allyl-6- (2-thiazolyl) acetoxymethyl-1,1-dioxopenicillanat

Acetylating 0.5 g (1.29 mmol) of allyl 6- (2-thiazolyl) -hydroxymethyl-1,1-dioxopenicillanate (from Example 18) with 0.396 g (3.88 mmol) of acetic anhydride and 0.307 g (3, 88 mmol) of pyridine in 5 ml of tetrahydrofuran was carried out according to the method of Example 20, Part A, with stirring at room temperature for 4 hours. The mixture was then diluted with methylene chloride, washed with water until neutral (pH 6> 0-6.5), the organic phase dried over Na ₂ SO _4, and the solvent evaporated to give 0.688 g of the desired acetate deliver. ¹ H-NMR _(CDCl3) ppm (X): 1.52 (s, 3H), 1.70 (s, 3H), 2.35 (s, 3H), 4.4-4.6 (m, 2H), 4.6-5.0 (m, 3H), 5.2-6.4 (m, 3H), 6.65 (d, 1H), 7.4 (d, 1H), 7.8 (d, 1H).

Example 36

Allyl-6- (2-thiazolyl) methylene-1, 1-dioxopenicillanate and its hydrolytes to the potassium salt

A. The above acetoxyester (0.688 g, 1.29 mmol) was mixed with 0.16 g (1.29 mmol) of 1,5-diazabicyclo / 4,3,7-non-5-ene (DBN) and 5 mL of methylene chloride The resulting mixture was diluted with methylene chloride, washed with water (2x50 mL), dried over Na ₃ SO _4, and the solvent was evaporated to give an oil, which was eluted on a silica gel column 1. Chromatograph 1 ethyl acetate / hexane to give 0.189 g (39%) • light yellow oil to yield ¹ H-NMR _(CDCl3) ppm {£): 1.53 (s, 3H), 1.65 (s, 3H), 4.33 (s, 1H), 4.55 (d, 2H), 5.0-5.4 (m, 2H), 5.45 (s, 1H), 5.4-6, 0 (m, 1H), 7.1 (m, 1H), 7.75 (m, 1H), 7.65 (d, 1H).

B. Hydrolysis of the allyl ester obtained above by the method of Example 25 Part C yielded potassium 6- (2-thiazolyl) -ethylene-1/1-dioxopenicillanate. in 84.7% step yield as a yellow solid. ¹ H-NMR 25OMHz fDMSO-Dg) ppm ( £) : 1.40 (s, 3H), 1.45 (s, 3H), 3.80 (s, 1H), 5.83 (s, 1H), 7.66 (s, 1H), 8.04 (rn, 2H).

Example 37

Tetrabutylammonium 6- / t> - (2- / 1-methyl-1-methoxycarbonylvinyl amino / 2-phenylacetamido) -penicillanate

To 300 ml of chloroform was added 39.3 g of 6- / D- (2-amino-2-phenylacetamido) -penicillanic acid trihydrate, 50 ml of water was added and the pH of the mixture was adjusted to 8.5 by addition of 40% aqueous tetrabutylammonium hydroxide set. The layers were separated, the aqueous layer saturated with sodium sulfate and extracted with fresh chloroform. The extracts and the initially deeper layer were combined and the solvent was evaporated to a total volume of about 250 ml.

To this was added 150 ml of methyl acetoacetate and 30 g of anhydrous magnesium sulfate. The mixture was heated at reflux for 3 hours, the mixture was allowed to sit and the warm organic layer was decanted. The clear chloroform solution was allowed to cool to give crystals of the title compound in 52% yield, mp 182-184 ° C (dec.). ¹ H-NMR _(CDCl3) ppm [S): 0.8-2.0 (m, 4H), 1.88 (s, 3H), 3.1-3.6 (m, 8H), 3, 6 (s, 3H), 4.17 (s, 1H), 4.58 (s, 1H), 5.05 (d, 1H), 5.38-5.6 (m, 2H), 6.78 (d, 1H), 7.35 (s, 5H), 9.4 (d, 1H).

Example 38

Tetrabutylammonium 6- / b- (2-1 * -methyl-methoxycarbonylvinyl amino 7-2- / 4-hydroxypheriyl-7-acetamido) ./periicillariat

To 300 ml of dichloromethane are added 41.9 g of 6- (2-amino-2- / 4-hydroxy

phenyl-7-acetamido) penicillanic acid trihydrate and 50 ml of water, and then the pH is adjusted to 8.5 with 40% aqueous tetrabutylammonium hydroxide. Three layers are obtained. The upper layer is removed, saturated with sodium sulfate and then extracted with dichloromethane. The extracts are combined with the middle layer and lower layer, and the resulting mixture is evaporated in vacuo to an oil which crystallized on rubbing with acetone. This provides 44.6 g of tetrabutylammonium 6- (2-amino-2 - / * 4-hydroxyphenyl / acetamido) -penicillanate.

The above salt is added to 150 ml of methyl acetoacetate and the suspension is heated to about 65 ^° C. until a clear solution is obtained (8 minutes). The mixture is allowed to cool and then the solid is recovered by filtration. The. Solid is washed with methyl acetoacetate then with diethyl ether to give AS, 25 g of tetrabutylammonium 6- (2- / "1-methyl-2-methoxycarbonyl-vinylamino / -2- /" 4-hydroxyphenylacetamido) penicillanate crystals.

Example 3 9 _v

A. Potassium (6- R, 8 S ) -6-pyrimidin-2-yl) hydroxymethyl- 1,1-dioxopenicillanate

To a solution of 300 mg (0.79 mmol) of the first-eluted isomer of allyl-6-c- (pyrimidin-2-yl) hydroxymethyl-1,1-dioxopenicillanate (contained in Example 18) in 4 mL of ethyl acetate Added 30 mg of tetrakis (triphenylphosphine) palladium (0) and 30 mg of triphenylphosphine. The mixture was stirred under nitrogen to solvate the reagents (5-10 min) and 1.57 ml (0.79 mmol) of potassium 2-ethylhexanoate in ethyl acetate were added. After stirring for 20 min at room temperature, the mixture was filtered and the cake was washed with ethyl acetate and dried to afford 53 mg of yellow solid. The filtrate was treated with ethyl ether to precipitate a second crop, 152 mg; Total-

yield 69%. ¹ H-NMR, ^ 250 MHz, (DMSO-dg) ppm (S) '. 1.33 (s, 3H), 1.44 (s, 3H), 3.77 (s, 1H), 3.95 (d of d, J = 2, J = 6, 1H), 4.89 ( d, J = 2, 1H), 5.1 (d, J = 6, 1H), 6.33 (s, 1H), 7.48 (t, J = 4, 1H), 8.84 (d, J = 4, 2H).

For example, potassium (6-U, 8R) -6- (pyrimidin-2-yl) hydroxymethyl-1,1-dioxopenicillanate

A solution of 300 mg (0.79 mmol) of the second-eluted isomer of allyl-6- (* pyrimidin-2-yl) hydroxymethyl-1,1-dioxopenicillanate (obtained in Example 18) was prepared according to the above procedure in the potassium salt was converted to 236 mg (79%). ¹ H-NMR, 250 MHz (DMSO-dg) ppm (S) ι 1.30 (s, 3H), 1.42 (s, 3H), 3.65 (s, 1H), 4.60 (dd, J = 2, J = 8, 1H), 4.75 (d, J = 2, 1H), 5.15 (d, J = 8, 1H), 7.47 (t, J = 4, 1H), 8.85 (d, J = 4, 2H).

Example 40

A. Allyl (6-o, 8S) -6- (pyrimidin-2-yl) acetoxymethyl-1, 1- dioxopenicillanate

To a solution of 785 mg (2.1 mmol) of the first-eluted isomer of allyl-6-r- (pyrimidin-2-yl) -hydroxymethyl-1,1-dioxopenicillanate (obtained in Example 18) in 4 mL of methylene chloride 0.45 m (5.6 mmol) of pyridine and 0.53 ml (5.6 mmol) of acetic anhydride were added and the mixture was stirred for 2.5 h at room temperature. The mixture was diluted with 30 ml of methylene chloride, extracted with water (7x60 ml), dried over anhydrous magnesium sulfate and filtered. Evaporation in vacuo gave 813 mg (92%) of the title compound. ¹ H-NMR _(CDCl3) ppm (£): 1.4 (s, 3H), 1.6 (s, 3H), 2.2 (s, 3H), 4.45 (s, .3H) - , 4.45 (dd, 1H), 4.75 (m, 2H), 4.95 (d, 1H), 5.2-5.6 (m, 2H), 5.7-6.3 (m , 1H), 6.45 (d, 1H), 7.35 (t, 1H), 8.85 (d, 1H).

Allyl (6- ", 8R) -6-pyrimidin-2-yl) acetoxymethyl- 1,1-dioxopenicillana"

Acetylation of the second-eluted isomer of allyl 6-egg- (pyrimidin-2-yl) hydroxymethyl-1/1-dioxopenicillanate (obtained in Example 18) by the above method gave 88% yield of the title compound. H-NMR _(CDCl3) ppm (S): 1.4 (s, 3H), 1.6 (s, 3H), 4.45 (s, 1H), 4.50 (dd, J = 1, J = 8, 1H), 4.75 (m, 2H), 4.8 (d, J = I, 1H), 5.25-5.6 (m, 2H), 5.7-6.3 (m , 1H), 6.4 (d, J = 8, 1H), 7.35 (t, J = 6, 1H), 8.8 (d, J = 6, 1H).

Example 41

A. Potassium (6oi, 8S) -6- (pyrimidin-2-yl) acetoxymethyl-1,1- dioxopenicillanate

A solution of 789 mg (1.86 mmol) of allyl (6-ei, 8S) -6- (pyrimidin-2-yl) acetoxymethyl-1,1-dioxopenicillanate in 4 ml of ethyl acetate was reacted according to the procedure of Example 54, to give 342 mg (43%) of the desired potassium salt, which was purified by preparative medium pressure liquid chromatography eluting with 9: 1 water / acetonitrile to give 105 mg of product (85% pure by high pressure liquid chromatography analysis).

For example, potassium (6- *, 8R) -6- (pyrimidin-2-yl) acetoxymethyl-1, 1- dioxopenicillanate

A solution of 666 mg (1.57 mmol) of allyl (6- te, 8R) -6- (pyrimidin-2-yl) acetoxymethyl-1,1-dioxopenicillanate was reacted by the same procedure to give 339 mg (51%). ) To give crude product which was purified by preparative medium pressure liquid chromatography (MPLC) with 9: 1 water / acetonitrile to afford 162 mg of pure isomer. H NMR, 250 MHz, (DMSO-d ₆ ) ppm (£): 1.34 (s, 3H), 1.44 (s, 3H), 2.17 (s, 3H), 3.65 fs, 1H), 4.15 (dd, J = 2, J = 8, 1H), 4.97 (d, J = 2, 1H), 6.27 (d, J = 8, 1H), 7.50 ( t, J = 5, 1H), 8.85 (d, J = 5.2H).

Example 42

.13,

Using the appropriate starting 6-R CHOH-subst, -1,1-dioxopenicillanate ester of Example 18, the following acetate esters are prepared by the method of Examples 20 or 55.

OCOCH 13 ' ^X -CH ^ IN 0 0 \ / I / ^CH 3 --CH ₃ ^ COOCH ₂ CH = CH ₂

13

S 5 \

* "6 ^f ^ 8

stereochemistry

and 6 tf,

8R mixture

Yield 100

60:40

Mixture of 6-flf / 8S 6 - # <, 8R

69

H-NMR (CDCl ₃ ) ppm (delta);

1.43, (s, 3H), 1.63 (s,

3H), 2.25 (s, 3H), 4.51

(m, 2H), 4.79 (m, 2H),

5.43 (m, 2H), 5.98 (m,

IH), 6.65 (d, IH), 7.5

(m, 3H), 7.98 (s, IH), 8.20 (m, 2H).

1.4 (s, 1.8H), 1.43 (s, 1.2H), 1.56 (s, 1.2H), 1.62 (s, 1.8H), 2.2 (s, 1.2H), 2.3 (s, 1.8H), 4.35 ( m, IH), 4.4 (s, 0.6H), 4.43 (s, 0.4H), 4.78 (d, 0.6H), 4.8 (d, 0.4H), 5.3-5.5 (m, 2H), 5.8-6.05 ( m, IH), 6.3 (m, IH), 7.45 (d, IH), 8.82 (d, IH), 9.25 (m, IH) '.

Example 42 (continued)

^C 6 ' ^C 8 ~

ι 7 stereo% ι

Chemistry Yield " ¹ H-NMR (CDCl ₃ ) PPm (delta):

€ 1

1.4 (s, 1.8H), 1.5 (s, ^ _x mixture of 1.2H), 1.6 Cs, 1.8H),

^VN SU ₁ 8S 1.65 (s, 1.2H), 2.2 (s,

6-o, 8R

1.2H), 2.26 (s, 1.8H),

4.23 (dd, 0.4H), 4.35 (dd, 0.6H), 4.4 (s, 0.6H), 4.45 (s, 0.4H), 4.68 (m, 2H), 4.74 (d, 0.6H), 5.0 (i , 0.4H), 5.35 (m, 2H), 5.9 (m, IH), 6.45 (m, IH), 8.6 (m, 2H), 8.75 (m, IH).

Example 43

The e-acetoxy-S-carbonyloxyallyl esters of Examples 25A and 57 are converted to the potassium salt of the following formula by the method of Example 54.

OCOCH

ti

Stereo-

65:35 6-tf, 8S S-öi, 8R

\\ 6: 1

^N 6- *, 8S

• 6-o £, 8R

yield

57

(raw)

21

isomeric

64 (D ₂ Q) ppm (delta)

purified: ι ₄₅ f-1.58 (s, 3H), 1.25 (s,

4.25 (s, 0.65H), 4.28 (s, 0.35H), 4.37 (dd, 0.65H), 4.45 (dd, 0.35H), 5.15 (d, 0.65H), 5.2 (d, 0.35H), 6.25 ( d, 0.65H), 6.35 (d, 0.35H), 7.73 (m, IH), 8.85 (m, IH),

9.15 (m, IH).

¹ ^ ⁴⁴ is / 3H), 1.5 ( _{S /} 3H), 1.62 (s, 3H), (s, 0.4H), 2.24 (s, 2.6H), 3.8 (s, 3H), 4.27 (s, IH) .

4.4 (dd, IH), 4.96 (d, IH), 6.45 (d, 0.15H),

6.5 (d, 0.85H), 7.07 (s, 0.15H), 7.1 (d, 0.85H),

7.16 (s, 0.15H), 7.2 (d, 0: 85H).

IR (KBr): 3409, 1786,

1740, 1620 cm

" ¹

Example 43 (continued)

^C 6 ' ^C 8-

Stereo%

R chemistry Yield _ H-NMR (DMSO-d _g ) ppm (delta):

30:70 84, χ. 3 ( _s , 2.1H), 1.34 (s,

6-S 8R ^ 43, Q.9H), 1.42 (s, 3H), 2.13

chromate ( _Sf o.9H), 2.2 (s, 2.1H),

crraphie

, 3.66 (s, 0.7H), 3.7 (s,

0.3H), 4.1 (dd, 0.3H), 4.95 (d, 0.7H), 5.07 (d, 0.3H), 6.24 (d, 0.3H), 6.36 (d, 0.7H), 8.7 (s, 2H) , 8.8 (s, 0.7H), 8.83 (s, 0.3H).

IR (KBr): 3468, 1781, 1746, 1623 cm " ¹ .

* C ^ g column uses TC .. _g is monooctadecylsilicate)

Example 44

The 8-acetoxy esters provided above and below are converted to the corresponding 6-methylene compounds by the method of Example 20, part B, and the allyl group removed by the method of Example 54 to provide the potassium salt of the following formula wherein R Potassium is.

0 0

CH ₃ CH ₃

'COOR ¹ '

R = allyl

^RJ

Kaliura

yield

yellow

powder

¹ H-NMR (CDCl ₃ ) Ppm (delta);

1.50 (s, 1.5H), 1.54 (s, 1.5H), 1.65 (s, 3Hj, 4.58 (s, IH), 4.80 (d, 2H), 5.30 (s, IH), 5.50 (m, 2H), 6.00 (m, IH), 7.10 (s, IH), 7.5 (m, 3H), 8.02 (s, 0.5H), 8.25 (m, 2H), 8.87 (s, 0.5H). A 1: 1 Mixture of E and Z isomers.

% Yield

42, hur E isomer

''"11-HMR (D ₂ O or d _fi -Acetom) ppm (delta):

1.53 (s, 3H), 1.63 (s, 3H), 4.3 (s, IH), 5.7 (s, IH), 7.2 (s, IH), 7.4-8.0 (m, 5H), 8.7 (s ,, " IH).

Example 44 (continued)

R ¹ = Allvl

Exploit e

(Parent compound, Ex. 68B)

(Parent compound, Ex. 68B)

46

59

1.48 (s, 3H), 1.62 (s,

3H), 4.52 (s, IH), 4.60-

4.80 (s, 2H), 5.31-5.44

(ro, 2H), 5.74 (s, IH),

5.87-6.01 (m, IH), 7.59

(s, IH), 8.76 (s, IH).

2.46 (s, 3H) ₁ 2.60 (s, 3H), 4.46 (s, IH), 5.20-5.42 (m, 2H), 5.66 (s, IH), 5.80-6.02 (m, IH), 7.38-7.42 ( m, 2H), 8.71 (m, IH).

Infrared; 1795 cm

-1 R ¹ = potassium

% Yield

isolated as a by-product from corre sponding compound in Ex. 58

H-NMR (D ₂ O or

) Ppm (delta);

39 yellow solid

3.5: 1 ratio of p: wp isomers

_ 1.55 (s, 3H), 1.65 (s, 3H)

4.35 (s, IH), 5.12 (d, IH)

7.52 (d, IH), 7.68 (d, Ih)

8.85 (m, IH), 9.2 (m, IH).

1:38 (S, cm 3H), 1 .40 (S, 3H), 4.19 (S, IH) 5 .92 (S, IH) 7.76 (S, IH) 9 .13 (S, III). Infrared. (KBr) * • 1775 1620 •

1.40 (s, 3H), 1.48 (s, 3H), 4.20 (s, IH), 5.91 (s, 1H), 7.40-7.60 (m, 2H), 8.89 (m, IH). Infrared (KBr): 1760, 1610 cm " ¹ .

Example 44 (continued)

R ¹ = allyl

Yield e ¹ II-NMR (CDCl ₃ ) ppm (delta):

95

starting from the 8-hydroxy compound of Example 18

1..52 (s, 3H) ₁ 1.64 (s, 3H) 2.46 (s, 3H), 4.5 (s, IH), 4.66-4.82 (m, 2H), 5:34 to 5:46 (m, 2H), 5.65 (s, IH), 6.0 (m, IH), 7.16 (s, IH), 7.34 (s, IH). Infrared: 1790, 1760, 1680 cm ' ¹ .

1.49 (s, 3H), 1.63 (s,

3H), 4.52 (s, IH), 4.60-

4.78 (m, 2H), 5.20-5.45 (m, 2H), 5.96-6.02 (m,

2H), 7.39 (s, IH), 7.48-

7.58 (m, 2H), 9.12-9.21 (m, IH).

R ¹ = potassium

84

20

"H NMR (D ₂ O or

% Yield d ₆ -acetone) ppm (delta)

1.38 (s, 3H), 1.46 (s, 3H), 2.33 (s, 3H), 4.16 (s, IH), 5.89 (s, IH), 7.3 (s, IH), 7.46 (s, IH).

Infrared (KBr): 1773, 1683, 1619 cm " ¹ .

Infrared; 1780, 1625 cm

Example 44 (continued)

CH.

R ¹ = allyl

Exploit e

64

-oil

H-NMR (CDCl ₃ ) ppm (delta):

1.46 (s, 3H), 1.60 (s, 3H), 3.64 (s, 3H), 4.51 (s, IH), 4.60-4.80 (m, 211), 5.24-5.46 (m, 2H), 5.62 (s, IH), 5.80-6.02 (m, IH), 7.32 (s, IH). Infrared: 1805 cm ".

R ¹ = potassium

% Yield

H-NMR (D ₂ O or d _fi -acetone) ppm (delta);

1.42 (s, 3H), 1.48 (s, 3H), 2.52 (s, 3H), 4.25 (s / IH), 5.87 (s, IH), 7.18 (s, IH). Infrared (KBr): 1785 cm " ¹ .

Example 45 Potassium 6-6-imidazol-2-yl) hydroxymethyl-1/1-dioxopeniculanate

A mixture of 141 mg (0.38 mmol) of allyl-6- (imidazol-2-yl) hydroxymethyl-1, 1-dioxopenicülanate / "isomer mixture, obtained in Example 187/12 mg of tetrakis (triphenylphosphine) palladium (0). , 12 mg of triphenylphosphine, 0.76 ml (0.38 mmol), potassium 2-ethylhexanoate and 2 ml of ethyl acetate was stirred under nitrogen for 1 hour The precipitated product was collected by filtration to give 143 mg (100%) of a yellow solid which was found to contain two isomers of high pressure liquid chromatographic analysis IR (KBr): 3382, 1780, 1728 and 1615 cm ^-1 '

Example 46

A. Benzyl 6- (2-thiazolyl) hydroxymethyl-1,1-dioxopeniculanate

A solution of 17.79 g (44 mmol) of benzyl 6-o-bromo-1,1-dioxopeniculanate in 250 ml of dry tetrahydrofuran was reacted with an equimolar amount of methylmagnesium bromide at -78 ° C, then after stirring for one min Equimolar amount of thiazole-2-carboxaldehyde was added and stirred for a further 10 min. An equimolar amount of acetic acid was added, and after stirring for 5 minutes, the mixture was poured into 500 ml of water. Extraction with ethyl acetate, washing the extracts with water, drying (MgSO 4) and evaporation of the solvent in vacuo gave 16.93 g (89%) of crude product which showed two spots at TLC. The crude product was purified by silica gel / column chromatography eluting with chloroform / ethyl acetate, 96: 4, to give 4.72 g of a more polar isomer, 2.98 g of a less polar isomer, and 0.5 g of a mixture of isomers (43% overall yield) result.

/1-1.85-31643

More polar isomer: ¹ H-NMR _(CDCl3) ppm (S): 1.25 (s, 3H), 1.55 (s, 3H), 4.3 (dd, 1H), 4.45 (s, 1H ), 4.65 (bs, 1H), 4.9 (d, 1H), 5.2 (m, 2H), 5.55 (d, 1H), 7.35 (m, 6H), 7.75 (d, 1H).

Less polar isomer: ¹ H-NMR _(CDCl3) ppm (<5) 1.2 (s, 3H), 1.5 (s, 3H), 4.35 (m, 2H), 4.75 (d , 1H), 5.1 (m, 2H), 5.55 (d, 1H), 7.2 (m, 6H), 7.6 (d, 1H).

Diphenylmethyl 6- (2-thiazolyl) hydroxymethyl-1,1-dioxopenicillanate

Repeating the above procedure with diphenylmethyl 6- oc- bromo-1,1-dioxopenicillanate on a 20 mm scale and purifying by silica gel chromatography eluting with 9: 1 chloroform / ethyl acetate gave 2.4 64 g of a less polar (wp) isomer and 3.029 g of a more polar (p) isomer.

More polar isomer: ¹ H-NMR _(CDCl3) ppm (S): 1.06 (s, 3H), 1.52 (s, '3H), 4.1-4.3 (m, 1H), 4, 42 (s, 1H), 4.76 (d, 1H), 5.45 (d, 1H), 6.82 (s, 1H), 7.05-7.3 (m, 11H), 7.56 (d, 1H).

Less polar isomer: ¹ H-NMR _(CDCl3) ppm (£): 1.2 (s, 3H), 1.65 (s, 3H), 4.35 (dd, 1H), 4.55 (s, 1H), 4.83 (d, 1H), 5.65 (dd, 1H), 6.95 (s, 1H), 7.2-7.4 (m, 11H), 7.75 (d, 1H )

Example 47

8 Using the appropriate acid chloride of the formula (R Cl or the corresponding

The following compounds were prepared according to the method of Example 20, Part A, of the following acid anhydride and the appropriate 6-R CHOH-subst.-1,1-dioxopenicillanate ester prepared above.

OR ¹⁸ OO

R ¹³ -CH

S /

H-

CH

COOR

R ^a C ₆ H ₅ CH ₂ R ¹³ R ¹⁸ Stereo chemistry aw Cq (from the more polar 8-hydroxy compound of Example 61A) 0 2-thiazolyl ( Il "Ht-C £ 3 (R) t

H-NMR (CDCl ₃ ) ppm (delta);

1.3 (t, 3H), 1.4 (s, IH), 1.65 (s, IH), 2.55 (q, 211), 4.5 (s, 111), 4.4-4.6 (m, IH), 4.85 (d, III) , 5.27 (s, 2H), 6.65 (d, 111), 7.35 (m, 6H), 7.75 (d, IH).

Example 47 (continued)

R ^c

(C ₆ H ₅ J ₂ CH

(from the less polar isomer of Example 61B)

(C ₆ H ₅ ) ₂ CH

(from the more polar isomer of Beisp.61B)

allyl

13

2-T. Thiazolyl C ₂ II ₅ OC

2-thiazolyl

2-thiazolyl

0 C ₂ H ₅ OC

stereochemistry

on the C _Q

(S)

(R)

¹ H-NMR _(CDCl3) ppm (delta);

1.25 (s, 3H), 1.3 (t, 3H), 1.75 (s, 3H), 4.24 (q, 2H), 4.36-4.60 (m, III), 4.55 (s, IH), 5.02 (d, IH) , 6.43 (d, IH), 6.95 (s, IH), 7.1-7.5 (m, HH), 7.75 (d, IH).

Pink, solid foam

1.4 (s, 2.1H), 1.5 (s, 0.9H), 1.6 (s, 2.1H), 1.65 (s, 0.9H), 4.4 (s, 0.7H), 4.5 (0.3H), 4.5-5.0 (m, 4H), 5.1-6.2 (m, 3H), 6.9 (d, IH), 7.4 (m, IH), (d, IH).

Example 47 (continued)

Stereochemistry R ^ R ^XJ R ^xo at C ₈ H-NMR (CDCl ₃ ) ppm (delta);

0 allyl 2-thiazolyl (CH ₃ J ₂ CHC (S) 1.37 (m, 3H), 1.50 (m, 3H),

polar? * ¹ ^ ^{65 (s} ' ^3H) 'i · ^82. (s » ^3H ) ' 22% 2.70 (in, IH), 4.61 (s / IH),

Yield. _, "". , _, _ "

4.7 (ra, IH), 4.9 (nt, 2H] ₁

5.12 (d, IH), 5.41 (m, IH), 5.6 (m, IH), 5.8-6.5 (m, IH), 6.75 (d, IH), 7.6 (d, IH), 7.97 (d, IH) ,

^ _(R) 1.18 (rn, 3H), 1.28 (m, 3H),

polar, 1.33 (s, 3H), 1.52 (s, IH),

/ "Yield ² * ^{6 (m} ' ^1H) ' ⁴ ^ ³ " ⁴ - ⁷ ^> ^5H ) >

5.0-5.4 (m, 2H)., 5.45-6.2 (m,

, IH), 6.47 (d, IH), 7.23 (d, IH), 7.63 (d, IH).

Example 47 (continued)

R ^c

C ₆ H ₅ CH ₂

R ¹³ R ¹⁸ Stereo chemistry O 2-thiazolyl Il (S) less polar (R) more polar

H-NMR _(CDCl3) ppm (delta):

1.4 (s, 3H), 1.6 (s, 3H), 4.5 (s, IH), 4.4-4.6 (m, IH), 5.1 (d, IH), 5.2 (s, 2H), .6.7 (d, IH ), 7.2-8.2 (m, 12H)

 1.3 (s, 3H), 1.6 (s, 3H), 4.5 (s, IH), 4.6 (dd, IH), 4.86 (d, IH), 5.2 (s, 2H), 6.85 (d, IH), 7.3 -7.7 (m, 9H), 7.8 (d, IH), 8.15 (dd, 2H).

Example 48

A. (6-f **, 8R) - ^ - (thiazol-2-yl) propionyloxymethyl 1-1,1-dioxopenicylic acid

A mixture of 1.89 g of 10% Pd / C catalyst in 20 ml of a 9: 7 (v / v) mixture of tetrahydrofuran (THF) and water was saturated with hydrogen and a solution of 689 mg (1.4 mmol) Benzyl- (6 - <*, 8R) -6- (thiazol-2-yl) -propionyloxymethyl-1,1-dioxopenicillanate in 13 ml of THF and 7 ml of water was added. The resulting mixture was hydrogenated at 3 bar pressure for 20 min, the catalyst was then filtered off, the filtrate extracted with ethyl acetate (3x200 ml) and the extracts dried (over MgSO 4). Evaporation of the solvent in vacuo gave 330 mg of yellow solid.

B. (6 - <* :, 8R) -6- (thiazol-2-yl) benzoyloxymethyl-1,1-dioxo penicillanic acid

The title compound was obtained by the above procedure from the corresponding benzyl ester in 57% yield. ¹ H-NMR (D ₂ O) ppm (£): 1.38 (s, 3H), 1.55 (s, 3H), 4.25 (s, 1H), 4.44 (dd, 1H), 5.05 (d, 1H), 6.68 (d, 1H), 7.4 (t, 7H), 7.55 (t, 1H), 7.58 (d, 1H), 7.7 (i.e. , 1H), 7.95 (d, 1H). IR \ KBr): 3473, 1782, 1729, 1622 cm " ¹ ,

Example 4 9

A. (6- ", 8S) -6- (thiazol-2-yl) ethoxycarbonyloxymethyl- 1,1-dioxopenicillanic acid

To a solution of 557 mg (0.954 mmol) of diphenylmethyl- (6-cxf, 8S) -6- (thiazol-2-yl) ethoxycarbonyloxymethyl-1, 1-dioxopenicillanate in 5 mL of methylene chloride was added 0.62 mL (5.72 mmol ) Given anisole. The mixture was cooled to -5 ° and a mixture of 382 mg (2.86 mmol) of anhydrous aluminum chloride and 2 ml of nitromethane slowly over 15 minutes

added. The reaction mixture was diluted with 50 mL of ethyl acetate, water added, and the pH adjusted to 7.5. The aqueous layer was separated, acidified to pH 3 and extracted with ethyl acetate. Evaporation of the solvent gave a residual glass, which was dissolved in ethyl ether, filtered, and hexane was added to the filtrate to cause precipitation. After filtering to give the solid and drying, 211 mg (53%) of product was obtained. ¹ H-NMR, 300 MHz, (CDCl ₃ ) ppm (S): 1.40 (t, 3H), 1.53 (s, 3H), 1.67 (s, 3H), 4.28-4, 42 (m, 3H), 4.50 (s, 1H), 4.92 (s, 1H), 6.58 (d, 1H), 7.53 (d, 1H), 7.94 (d, 1H ). IR (KBr): 3443, 1797, 1754 cm " ¹ .

B. Use of the (6-, r, 8R) isomer of the starting diphenylmethyl ester prepared in Example 62 in the above procedure affords the corresponding (6- *, 8R) isomer of 6- (thiazol-2-yl) ethoxycarbonyloxymethyl 1,1-dioxopenicillanic acid. ¹ H-NMR, 300 MHz, (CDCl ₃ ) ppm (S): 1.34 (t, 3H), 1.53 (s, 3H), 1.65 (s, 3H), 4.2-4, 4 (m, 3H), 4.44 (s, 1H), 5.04 (s, 1H), 6.67 (d, 1H), 7.53 (d, 1H), 7.90 (d, 1H ). IR (KBr): 3418, 1803, 1750 cm " ^1. "

Example 50

Using the appropriate allyl ester starting material prepared in Example 62 as the starting material, the following potassium salts were obtained by the method of Example 60.

OR "R ¹³ -CH.

, CH. CH.

'"COOK

2-thiazolyl

stereochemistry

(R)

¹ H-NMR (D. ₂ O) ppm (delta)

1.46 (s, 3H), 1.65 (s, 3H), 4.35 (2s, IH), 5.2 (d, 111), 7.05 (d, IH) , 7.7-8.0 (m, 211), 8.7-9.5 (m, 3H).

Example 50 (continued)

18

2-thiazolyl

(CH ₃ J ₂ CHC

Stereochemistry at C,

-Q-

(S) 90%

yield

(R) 27%

yield

¹ H-NMR (DMSO-d ₆ ) Ppm (delta);

1.13 (d, 3H), 1.16 (d, 3H), 1:33 (s _f 3H), 1:45 (s, 3H), 2.65 (m, IH), 3.73 (s, IH), 4.23 (dd, IH), 4.93 (d, IHj, 6.50 (d, IH), 7.85 (m, 2H) Infrared (KBr): 3777, 3472, 3447, 3402, 3270, 1783, 1746 1621 cm ^-1 .

1.17 (d, 3H), 1.20 (d, 3H), 1.44 (s, 3H), 2.69 (m, IH), 3.68 (s, IH), 4.12 (dd, IH), 4.93 (d, IH), 6.59 (d, IH), 7.84 (m, 2H).

Infrared (KBr): 3543, 3498, 3437, 3415, 3348, 3270, 3119, 1917, 1783, 1745, 1718, 1619 cm " ¹ .

Example 51

A. Allyl 6-bromo-e- (2-thiazolyl) hydroxymethyl-1,1-dioxo penicillanate

A solution of 8.84 g (20 mmol) of allyl-6,6-dibromo-1,1-dioxopenicillanate in 100 ml of dry tetrahydrofuran was cooled to -78 ° C, 7.02 ml (20 mmol) of methylmagnesium bromide were added and the Mixture stirred for 5 min. A solution of 2.26 g (20 mmol) of thiazole-2-carboxaldehyde in 10 ml of the same solvent was added at -78 ° C and the resulting mixture was stirred for 20 min. Acetic acid (1.2 ml) was added, the mixture was poured into water and extracted with ethyl acetate and chloroform. The combined organic layers were dried (over Na 2 SO 4 ) and the solvent removed in vacuo to give 8.5 g of crude product as a glass. The crude glass was purified by column chromatography on silica gel eluting with 89:11 chloroform / ethyl acetate to afford 6.2 g (72%) of pure product, which proved to be a single isomer. ¹ H-NMR _(CDCl3) ppm (S) '. 1.4 (s, 3H), 1.6 (s, 3H), 4.0 (bs, 1H), 4.42 (s, 1H), 4.6 (d, 2H), 5.3 (s , 1H), (5.55 s, 1H), 5.1 to 6.3 (m, 3H), 7.35 (d, 1H), 7.75 (d _f 1 H).

B. Benzyl-6-bromo-6- (2-thiazolyl) hydroxymethyl-1, 1-dioxo penicillanat

Use of benzyl 6,6-dibromo-1,1-dioxopenicillanate in place of the allyl ester in the above procedure afforded the title compound as an orange-colored foam in quantitative yield. ¹ H ₁ -NRR (CDCl 3) ppm (<T): 1.32 (s, 3H), 1.60 (s, 3H), 4.5 (s, 2H), 5.2-5.8 ( m, 4H), 7.3 (d, 1H), 7.4 (s, 5H), 7.8 (d, 1H).

Example 51 (continued)

C. The following compounds were prepared as in the procedure of Part A.

OH

0 0

CH.

• Ν-

Y-3

COOR "

R ^a R ¹³

benzyl

benzyl

Benzyl N, N

.N.

Allyl S '_ ^

% Yield ¹ H-NMR (CDCl3) ppm (delta);

100

yellow foam 1.25 (s, 3H) ₇ 1.54 (s, 3H), 4.5 (s, IH), 5.02 (s, 2H), 5.43 (s, IH), 5.68 (s, IH), 7.32 (m, 7H) , 8.00 (m, 2H).

88

yellow. , , _

Solid ⁴ · ° < ^s ' ^lH >'

1.4 (2s, 3H), 1.6 (s, 3H),

.9 (2, IH), 5.18 (m, 2H), 5.5-5.8 (m, 2H), 7.5 (s, 5H), 8.0 and

8.5 (2s, IH).

1.25 (s, 3H), 1.52 (s, 3H), 4.45 (s, 0.75H), 4.52 (s, 0.25H), 5.1-5.4 (m, 4H), 7.3 (s, 5H), 8.6 (m, 2H), 9.0 (m, IH).

1.38 (s, 3H), 1.60 (s, 3H),

4.38-4.73 (m, 3H), 5.0-6.0

(m, 5H), 7.10 (s, IH) _r 8.51 (s, IH).

Example 52

A. Acylation of the compounds provided in the previous example by the method of Example 57 or 62 gave the following compounds in a similar manner.

OR

.IB "

-CH /

br

0 0

A. Where R ^{a is} allyl and R ^{18 is} CH ₃ CO:

13

2-thiazolyl

% Yield 49

colorless crystals

'''COOR ³

¹ H-NMR (CDCl ₃ ) ppm (delta):

1.4 (s, 3H), 1.6 (s, 3H), 2.25 (s, 3H), 4.45 (s, IH), 4.65 (m, 2H), 5.4 (s, IH), 5.2-6.3 (m, 3H) .

6.7 (s, IH), 7.4 (d, IH),

7.8 (d, IH).

¹³ C-NMR (CDCl ₃₎ ppm (delta); 18.4, 19.8, 20.4, 60.7, 63.1, 64.4, 66.5, 66.9, 73.3, 120.3, 120.4, 130.6, 143.6, 163.7, 165.5 166.2, 168.6.

(+ 21% isomer ^{1 '40 <s' 3H)'} 1.60- <s,

Product)

3H), 2.25 (s, 3H), 4.48-

4.70 (m, 3H), 5.2-6.2 (m,

5H), 6.92 (s, IH), 8.83 (s, IH).

Example 52 (continued) B. Where R ^{a is} benzyl and R ^{18 is} CH 3 CO:

13

% Yield

¹ H-NMR (CDCl I ppm (delta):

2% iazolyl 100 1.25 (s, 3H), 1.55 (s, 3HI,

⁰¹ ^ f ₁ - 4.50 (s, Q.9Hl, 4.55 (s,

Isomer mixture ο.IH), 5.2 (m, 2H), 5.40

(s, 0.9H), 5.57 (s, 0.1H), 6.42 (s, O.lH), 6.60 Cs, 0.9H), 7.3 (m, 6H), 7.75 (d, IH).

N, N

^ ^N ^

100

yellow-orange foam

1.3 (s, 3H), 1.52Cs, 3H), 2.25 (s, 3H), 4.54 (s, IH), 5.02 (s, 2H), 5.52 (s, IH),

6.8 (s, IH), 7.3 (m, 7H),

7.9 (m, 2H).

1.25 (s, 3H), 1.5 (s, 3H), 2.19 (s, 3H), 3.95 (s, 3H),

4.4 (s, IH), 5.15 (s, 2H), 5.45 (s, IH), 6.3 (s, IH), 7.35 (s, 5H), 7.73 (s, IH).

90 3: 1-1.25 (s, 2.25H), 1.35 (s,

Isomer mixture 0.75H), 1.4 (s, 2.25H), 1.42 (s, 0.75H), 2.2 (s, 2.25H), 2.3 (s, 0.75H), 4.45 (s, 0.75H), 4.55 (s, 0.25 H), 5.1-5.3 (m, 2H), (s, 0.75H), 5.6 (s, 0.25H), 6.25 (s, 0.25H), 6.4 (s, 0.75H), 7.3 (m, 5H), 8.67 (m, 2H), 9.67 (s, IH).

Example 52 (continued)

C. Alternatively, the compounds of the above formula are prepared by the method of Example 6 6 with acylation of the reaction mixture prior to isolating the product by the following general method:

To a solution of 1.0 equivalent of 6,6-dibromo-penicillanatester in tetrahydrofuran at -78 ⁰ C was added 1.3 equivalents of methylmagnesium bromide dissolved in the same solvent, and the mixture was stirred for 15 min. The appropriate aldehyde (R CHO), 1.3 equivalents, in the same solvent was added at -78 ⁰ C to -68 ⁰ C and the reaction stirred for 30 to 60 min. Then 1.3 equivalents of acetyl were! Chloride added with stirring at -78 ⁰ C for 10 min was continued, and the product was then isolated by poured into ice / water, extracted with ethyl acetate, dried and the solvent in vacuo was deducted.

Where R ^{18 is} CH CO: ^ a- "13

benzyl

allyl

^C 6 ^H 5

Or.

% Ausbeut ^X e H-NMR _(CDCl3) ppm (delta)

100

77

oil oil

pale yellow foam

1.35 (s, 3H), 1.56 (s,

3H), 2.22 (s, 3H), 4.42

(s, IH), 4.60-4.74 (m,

2H), 5.24-5.42 (m, 3H),

5.79-5.96 (m, IH), 6.52

(s, IH), 7.32-7.34 (d,

IH), 8.70-8.74 (d, IH). Infrared: 1810,

1760 cm

-1

Example 52 (continued)

R ^a R ¹³

allyl

benzyl

benzyl

CH.

N ^ S

CH.

% Yield e ¹ H-NMR (CDCl ₃ ) .ppm (delta)

1.42 (.s, 3H), 1.62 (s,

yellow oil _3H ), 2.28 (s, 3H), 2.48 (s, 3H), 4.5 fs, IH), 4.S-4.8 (m, 2H), 5.28-5.47 (m, 2H), 5.82-6.0 ( m, IH), 6.3 (s, IH), 6.97 (s, IH)

Infrared .: 1810, 1760, 1730 cm " ¹ .

1.6 (s, 3H), 1.8 (s,

3H), 2.2 (s, 3H), 2.3 (s, 6H), 4.4 (s, IH), 5.2 (d, 2H), 5.3 (s, IH), 6.4 (s, IH), 7.3 (s, 5H ).

1.22 (s, 3H), 1.5 (s,

3H), 2.18 (s, 3H), 2.42

(s, 3H), 4.5 (s, IH), 5.16-5.36 (m, 3H), 6.18

(s, IH), 6.48 (s, IH), 7.4 (s, 5H).

on -. ft.q.il i 6432

Example 52 (Continued)

R ^a R ¹³ % Yield ¹ H-NMR (CDCl ₃ ) .ppm (delta)

Benzyl N ^ S 44 1.26 (.s, 3H), 1.5 (s,

/ yellow foam ". _ _ _. ". _.

) ' ^J 3H), 2.2 Cs, 3Hl, 2.4

CH ₃ (S ₁ 3H), 4.4 (s, IH),

5.16 (d, 2H), 5.3 (s, IH), 6.6 (s, IH), 6.8 (s, IH), 7.3 (s, 5H).

Allyl N- ^ N 36 1.40 (s, 3H), 1.60 (s,

N ₀ Jf / foam

2.65 (s, 3H), 4.20-4.8 (m, 3H), 5.1-6.2 (m, 4H), 6.41 (s, IH)

Infrared: - 1815, 1760 cm " ¹ .

OQ- '' 'i h. 4-. 1 / _

Example 53

A. Benzyl-6-β- (thiazol-2-yl) acetoxymethyl-1,1-dioxopenicillanate

To a solution of 74.6 g (134 mmol) of benzyl 6-bromo-6- (thiazol-2-yl) acetoxymethyl-1,1-dioxopenicillanate in 850 ml of benzene was added 43.99 g (151.2 mmol) of tri -n-butyltin hydride. The mixture was heated at reflux for 5.5 hours and allowed to stand overnight. The solvent was removed in vacuo, the residue taken up in hexane and extracted with acetonitrile (2x250 ml). The acetonitrile layer was evaporated, the residue slurried in ethyl ether, filtered and the cake washed with ether to give 33.28 g of colorless crystals. An additional 2.8 g was obtained by evaporation of the filtrate to dryness. The residue was taken up in benzene and 10 g of tri-n-butyltin hydride were added. The mixture was refluxed for 1 h and worked up as in the first crop; combined yield 56.3%.

The above first crop was purified by column chromatography on silica gel, eluting with 9: 1 chloroform / ethyl acetate. The product fractions were concentrated, slurried with 4: 1 ethyl ether / ethyl acetate, filtered, washed with ether to afford 22.6 g of white solid. ¹ H-NMR _(CDCl3) ppm (£): 1.25 (s, 3H), 1.53 (s, 3H), 2.1 (s, 3H), 4.58 (s, 1H), 4 , 80 (d, 1H), 5.2 (dd, 1H), 5.22 (g, 2H), 6.75 (d, 1H), 7.35 (s, 5H), 7.4 (d, 1H), 7.8 (d, 1H). ¹³ C-NMR _(CDCl3) ppm (6): 17.7, 19.9, 20.5, 54.5, 63.06, 63.6, 63.8, 64.5, 68.1, 121 , 8, 128.8, 128.9, 134.3, 142.6, 164.6, 166.5, 169.2, 170.5.

Example 53 (continued)

B. The following compounds were similarly obtained by debromination of the remaining compounds of Example 6-7.

13

OCOCH-

- ^CH \

benzyl

benzyl

N-N

benzyl

% AusbB \ i te ¹ H-NMR _(CDCl3) ppm (delta);

39

72

461.3 (s, 3H), 1.55 (s, 3H), 2.19 (s, 3H), 4.5 (s, IH), 4.79 (d, IH), 5.2 (m, 3H), 6.79 (d, IH), 7.32 (m, 7H),

7.9 (m, 2H).

1.29 (s, 3H), 1.55. (s, 3H),

2.10 (s, 3H), 4.03 (s, 3H), 4.5 (s, IH), 4Λ78 (d, IH), 4.'87 (dd, IH), 5.25 (q, 2H), 6.53 (d, IH ), 7.38 (s, 5H), 7.89 (s, IH).

1.3 (s, 3H), 1.55 (s, 3H), yellow solid-4.45 _{(s # lH) f Am6} _ ₅ ^ _{ - «toff

2H), 5.2 (m, 2H), 6.6 (dd, IH), 7.35 (s, 5H), 8.6 (ro, 2H), 8.83 (m, IH).

Example 53 (continued)

13

allyl

% Yield

70 oil "H-NMR (CDCl" _{2 "} ) ppm (delta):

1.44 (s, 3H), 1.62 (s> 3H), 2.10 (s, 3H), 4.51 (s, IH), 4.60-4.80 (m, 2H), 4.89-4.91 (d, IH), 5.26-5.42 ( m, 3H), 5.86-5.99 (m, IH), 6.88-6.92 (d, IH), 8.82 (s, IH)

 Infrared: 1795, 1750.

allyl

51 (wp isomer)

11 (ρ isomer) v / p isomer: 5.84-6.00 (m, IH), 6.28-6.42 (m, 2H), 6.62-6.77 (d, IH), 7.38-7.48 (d, IH), 8.64-8.70 ( d, IH). White Crystals, Infrared (KBr): 1807, 1760 cm ^-1 .

allyl

CH

B enyl

31 yellow oil

Mp.

CH CH

^CH 3 ^P 3 195.5 ⁰ C 1.46 (s, 3H), 1.64 (s, 3H), 2.14 (s, 3H), 2.48 (s, 3H), 4.5 (s, IH), 4.6-4.9 (m, 3H), 5.2-5.26 (dd, IH), 5.3-5.6 (m, 2H), 5.86-6.1 (m, IH), 6.7 (d, IH), 7.0 (s, IH).

Infrared: 1810, 1760 cm.

1.3 (s, 3H), 1.58; (s, 3H), 2.12 (s, 3H), 2.36 (d, 6H), 4.5 (s, IH), 4.75 (d, IH), 5.2-5.4 (dd, 3H), 6.6 (d, IH), 7.4 (s> - 5H).

· R- ι.

Example 53 (continued)

13

benzyl

benzyl

CH.

allyl

% Yield ¹ H-NMR _(CDCl3) ppm (delta):

16 1.28 (s, 3H), 1.55 (s, ~ 3H),

colorless festive,,

² · ^{λ (s} ' ^3Η) ' ² · ^{4 Cs} ' ^3H >'

single 4.5 (s, IH), 4.8 (dd, IH),

5.25 (q, IH), 6.15 (s, IH), 6.5 (d, IH), 7.4 (s, 5H).

43

CH

44 3: 1

Isomeric mixture 1.3 (s, 3H), 1.58 (s, 3H), 2.14 (s, 3H), 2.48 (s, 3H), 4.52 (s, IH), 4.8 (d, IH), 5.16-5.36 (AB quadruplet and dd, 3H), 6.7 (d, IH), 7.0 (s, IH), 7.4 (s, 5H).

1.37-1.40 (d, 3H), 1.58-1.60 (d, 3H), 2.12-2.14 (d, 3H), 2.58 (s, 3H), 4.48 (s, IH), 4.58-4.88 (m, 4H), 5.24 x 5.46 (m, 2H), 5.82-6.00 (πι, IH), 6.64-6.67 (d, 0.75H), 7.05-7.08 (d, 0.25H).

Infrared: 1810, 1765 cm

-1

Example 54

A. The benzyl esters prepared above were converted to the corresponding carboxylic acids of the following formula wherein R ^{b is} H by hydrogenation over Pd / C catalyst according to the method of Example 63.

OR R ¹³ -CH.

CH. CH.

In front"

, R

13

2-thiazolyl

18

0 CH ₃ C

% Yield

white solid mp 145-155 ° C (decomp.)

H-NMR (DMSO-d ₆ or D ₂ O) ppm (delta):

1.45 (s, 3H), 1.53 (s, 3H), 2.10 (s, 3H), 4.48 (s, IH), 5.0 (dd, IH), 5.41 (d, IH), 6.59 (d, IH), 7.91 (m, 2H).

¹³ C NMR: 17.1036, 19.3736, 20.2771, 53.2954, 62.5971, 62.8347 63.3676, 64.5698, 122.6881, 142.4252, 164.3975, 167.8111, 168.7027, 170 * 8035.

Example 54 (continued)

13

q O-

CH _n C

CH ₃ C

% Yield ¹ H-NMR (DMSO-d ₆ or D ^ Q) ppm (delta);

1.53. (S, 3H), 1.65 (s, 3H), 2.17 (s, 3H), 4.54 (s, IH), 4.87 (d, IH), 5.13 (dd, IH), 6.82 (d, 2H), 7.88 (d, IH), 8.07 (d, IH), 9.20 (bs, IH).

¹³ C-NMR: 17.6, 20.09, 20.5, 54.5, 63.2, 63.6, 63.9, 65.5, 121.9,

123.4, 126.1, 126.6, 135.3, 151.6, 166.6, 169.2, 169.3, 170.6.

(D ₂ O) 1.45 - (s, 3H), 1.57 (s, 3H), 2.15 (s, 3H), 4.33 (s, IH), 5.24

(d, IH), 4.75-5.2 (-IH blocked by the D ₂ O-Peakl, 6.55 (d, IH), 8.65-8.8 (m, 2H) Infrared (KBr): 3416, 1785, 1618 cm ^{-1 13} C-NMR: 17.6, 19.8, 20 .. 4, 54.4, 64.1, 65.7, 68.1, 144.2, 145.2,

145.5, 151.2, 172.7, 172.8.

Example 54 (continued)

% Yield ¹ H-NMR (DMSO-d _G or D _n Q) ppm (delta):

  , ii.ii Ii Q * £

1.48 (s, 3H), 1.55 (s, 3H), 2.15

(s, 3H), 2.28 (s, 3H), 2.32 (s, 3H), 4.3 (s, IH), 4.85 (dd, IH), _and 5.2 (d, IH), 6.6 (d, IH). Infrared (KBr): 1787, 1626, 1618 cm " ¹ .

1.48 (s, 3H)., 1.6 (s, 3H), 2.2 (s,

3H), 2.46 (s, 3Hl, 4.35 (s, IH), 5.2 (d, IH), 6.4 (s, IH), 6.55 (d, IH) Infrared (KBr): 1791, 1692, 1631 Cm ' ¹ .

' S CH ₃ C 57 1.5 (s, 3H), 1.6 (s, 3H), 2.2 (s,

3H), 2.45 (s, 3H), 4.35 (s, IH),

CH ₃ 4.9 (dd, IH), 5.25 (d, IH), 6.7

: ... (d, IH), 7.3 (s, IH). Infrared

(KBr): 1787, 1657, 1626 cm " ¹ .

R ¹³ R ¹⁸ 0 • j 0 CH ₃ C Il S ^ N CH ₀ C H 3 CH ₃ CH ₃ CH ₃ H = J

Example 54 (continued)

B. 6-.beta .- (Thiazol-2-yl) acetoxymethyl-1,1-dioxopenicillanic acid obtained in Part A above was prepared by treating an aqueous slurry of the acid with an equimolar amount of potassium bicarbonate in water and purifying by medium pressure liquid chromatography on a C " Column (C. "= octadecylsilicate) with elution with 9: 1 water / acetonitrile to obtain the corresponding potassium salt in 60% yield. ¹ H-NMR (DMSO-dg) ppm (<£ *): 1.37 (s, 3H), 1.48 (s, 3H), 2.07 (s, 3H), 3.80 (s, 1H ), 4.92 (dd, 1H), 5.12 (d, 1H), 6.55 (d, 1H), 7.89 (m, 2H). IR (KBr): 3454, 1788, 1630 cm " ¹ .

Example 55

A. 6- (Benzothiazol-2-yl) methylene-1,1-dioxopenicillanic acid, mixture of E and Z isomers

To a solution of 400 mg (0.91 mmol) of 6- (benzothiazol-2-yl) acetoxymethyl-1/1-dioxopenicillanic acid in 5 ml of water was added a solution of 0.15 g (1.82 mmol) of sodium bicarbonate in 2 ml Water was added and the resulting mixture was stirred for 2 h. The reaction mixture (pH 7.55) was freeze-dried. The lyophilizate was taken up in 8 ml of water, adjusted to pH 3.5 with dilute hydrochloric acid, extracted with ethyl acetate, the organic layers dried (over MgSO 4) and the solvent evaporated in vacuo. The resulting product was found to be a 60:40 mixture of (E) and (Z) isomers according to its NMR spectrum. ¹ H-NMR _(CDCl3) (ί <) ppm: 1.55 (s, 1.2H), 1.6 (s, 1.8H), 1.65 (s, 1.2H), 1.67 (s, 1.8H), 4.55 (s, 0.6H), 4.58 (s, 0.4H), 5.38 (s, 0.4H), 5.74 (s, 0.6H ), 7.44 (s, 0.4H), 7.48 (s, 0.6H), 7.5 (m, 4H), 7.89 (d, 1H), 8.07 (d, 0, 4H), 8.15 (d, 0.6H).

B. Similar treatment of the appropriate 6-R-CH (OAc) -subst.- Λ, 1-dioxopenicillanic acid, prepared as above, provided the compound of the following formula as a mixture of (E) - and (Z) isomers. 0 °

R -CHv. ^ »- S · ^ / 3

O ^ '' COOH

13 ^ 1

Y Yield 1 H NMR (D _n O) PDm (delta);

⁸ * ^ · κ ¹ ^ ^{54 (S} ' ^3H) ' ¹ ^ ^{62 (s} ''

: 40-Genusch _nA .

(E) and ⁴ - ⁰⁴ < ³ ' ^3H K 4.35 (s, IH),

(Na-SaIz) (Z) isomer 5.7 (s, 0.6H) , 5.95 (s,

0.4H), 7.26 (s, 0.6H), 7.55 (s, 0.4H), 8.09 (s, 2H).

Example 56 *

A. Quinium 6-bromo-6- (thiazol-2-yl) acetoxymethyl-1,1- dioxopenicillanate

Reacting 96 mg (0/2 mmol) of allyl-o-bromo-e- (thiazol-2-yl) acetoxymethyl-1,1-dioxopenicillanate (prepared in Example 67) by the method of Example 60 for 10 min and working up, as described, 46 mg (48%) provided yellow, solid product. ¹ H NMR (D ₂ O) PPm (<£): 1.45 (s, 3H), 1.6 (s, 3H), 4.4 (s, 1H), 5.55 (s, 1H) , 6.85 (s, 1H), 7.72 (d, 1H), 7.86 (d, 1H).

For example, potassium 6-bromo-6- (thiazol-2-yl) hydroxymethyl-1,1- dioxopenicillanate

Similarly, reaction of 220 mg of allyl 6-bromo-6- (thiazol-2-yl) hydroxymethyl-1,1-dioxopenicillanate (prepared in Example 66) by the above method for 20 minutes gave a 52% yield of the title salt as pale yellow solid. ¹ H-NMR (DMSO-dg) ppm (£): 1.35 (s, 3H), 1.47 (s, 3H-), 3.75 (s, 0.4H), 3.83 (s, 0.6H), 5.3 (d, 0.4H), 5.32 (d, 0.6H), 5.45 (s, 0.6H), 5.5 (s, 0.4H), 7 , 6-8.0 (m, 2H). IR (KBr): 3442, 1794, 1633 cm " ¹ .

Example 57

Potassium (6-β, 8S) -6- (thiazol-2-yl) -hydroxymethylpenicillanate A. Allyl-6-bromo-6- (thiazol-2-yl) hydroxymethylpenicillanate

To a solution of 9.971 g (24.99 mmol) of allyl-6,6-dibromopenicillanate in 150 mL of dry tetrahydrofuran, cooled to -78 ° C under nitrogen, was added 8.77 mL (24.99 mmol) of methylmagnesium bromide in THF and the mixture stirred for 15 min. The solution of 2.824 g (24.99 mmol) of thiazole-2-carboxaldehyde in 5 ml THF was added and the mixture was stirred again for 20 min at -78 ⁰ C. The reaction was by addition of

Quenched 1.43 ml (24.99 mmol) of glacial acetic acid, the mixture stirred for min, then allowed to warm to room temperature. It was then poured into water, extracted with 2x250 ml of ethyl acetate, the extracts washed with water (2x250 ml), dried over MgSO4, and the solvent evaporated in vacuo to give 10.36 g of orange oil. The oil was purified by column chromatography on silica gel eluting with 9: 1 chloroform / ethyl acetate to give 4.54 g of yellow solid (mixture of isomers) and 0.443 g of yellow foam, only more polar isomer (overall yield 4-6%). For the yellow foam: ¹ H-NMR _(CDCl3) ppm [S): 1.56 (s, 3H), 1.76 (s, 3H), 4.60 (s, 1H), 4.7 (m , 2H), 4.9-6.4 (m, 6H), 7.45 (m, 1H), 7.8 (m, 1H).

B. allyl-6-β- (thiazol-2-yl) hydroxymethylpenicillanate

The more polar isomer of Part A, 200 mg (0.462 mmol), was dissolved in 1 mL of benzene and 0.183 mL (0.693 mmol, 1.5 eq.) Of tri-n-butyltin hydride in benzene was added. The mixture was heated at reflux for 3 hours and allowed to stand at room temperature overnight. The solvent was removed in vacuo, the residue taken up in acetonitrile, washed with hexane and evaporated to a small volume, which was applied to a silica gel column and eluted with chloroform to give 73 mg (45%) of the desired product. ¹ H-NMR _(CDCl3) ppm (S): 1.65 (s, 3H), 1.87 (s, 3H), 3.8-4.4 (m, 1H), 4.05 to 4, 3 (dd, 1H), 4.65 (s, 1H), 4.78 (m, 2H), 5.3-5.6 (m, 2H), 5.6-6.3 (m, 3H) , 7.45 (m, 1H), 7.85 (m, 1H).

C. The product obtained in Part B, 73 mg (0.206 mmol), was converted to the potassium salt by the method of Example 60 to provide 58 mg (80%) of the title compound as a yellow solid. ¹ H-NMR, 300 MHz, (D ₂ O) ppm (S): 1.36 (s, 3H), 1.55 (s, 3H), 4.13 (dd, 1H), 4.18 (s , 1H), 5.32 (d, 1H), 5.41 (d, 1H), 7.56 (d, 1H), 7.60 (d, 1H).

Example 58

.13.

A. Use of the appropriate aldehyde, R CHO, in place of thiazole-2-carboxaldehyde in the procedure of Example 72, part A, similarly provided the corresponding compounds of the following formula.

OH R ¹³ -th.

% Healed

20

Isomer A (white crystals)

13

Isomer B (yellow oil) ^ COOCH ₂ CH = CH ₂

H-NMR (CDCl ₃ ) ppm (delta):

Isomer A: 1.46 (s, 3H), 1.64 (s, 3H), 4.42 (d, IH), 4.50 (s, IH), 4.64 (m, 2H), 5.26-5.50 (m, 3H), 5.84 ( s, IH), 5.8-6.0 (m, IH), 8.58 (d, 2H), 8.9 (s, IH),

¹³ C-NMR (CDCl _3): 26.4, 32.5, 64.4, 66.1, 69.8, 70.9, 73.7, 74.9, 119.6, 131.0, 143..0,

144.3, 144.6, 152.1, 166.8, 167.8 ppm.

Isomer B: 1.45 (s, 3H), 1.63 (s, 3H), 4.56 (s, IH), 4.65 (m, 2H), 5.1-5.5 (m, 4H), 5.6-6.0 (m, IH), 5.91 (s, IH), 8.57 (m, 2H), 8.78 (m, IH). ¹³ C-NMR _(CDCl3): 26.27, 32.57, 64.30, 66.19, 69.94, .70.77, 72.38, 74.73, 119.70, 130.98, 143.30, 144.57, 144.70,

152.04, 166.99, 167.99 ppm.

Example 58 (continued)

13

"• Ν

Cf-Ης

, 6 5

N.N

% Yield

25

(Isomer A / 8S)

40

(Isomer B, 8R)

83

Foam ¹ H-NMR (CDCl ₃ ) ppm (delta):

1.52 (s, 3H), 1.70 (s, 3H),

4.64 (s, IH), 4.72 (m, 2H),

4.86 (d, IH), 5.34-5.48 (m,

3H), 5.91 (s, IH), 5.92-6.05

(m, IH), 7.39 (t, IH), 8.87 (d, 2H).

1.52 (s, 3H), 1.71 (s, 3H), 4.61 (s, IH), 4.72 (m, 2H), 4.98 (d, IH), 5.30-5.52 (m, 3H), 5.90-6.04 (m, IH), 6.10 (s, IH), 7.40 (t, IH) ,. 8.85 (d, 2H).

1.49 (s, 3H), 1.69 (s, 3H), 3.56 (d, 0.7H), 3.89 (d, 0.3H), 4.7 (m, 3H), 5.5 (m, 3H), (m, 2H), 7.53 (m, 3H), 8.14 (m, 3H).

97

(crude isomer mixture)

Example 58 (continued)

B. Debromination of the above compounds by the method of Example 72, Part B gave the following compounds. ·. ^: - ^y OH

R ¹³ - ^ H _V C ^CH -3 0 ¹ NL ^ OOCH ₂ CH = CH ₂ R ¹³ % Yield ¹ H NMR (CDCl ₀ ) ppm (delta): 57 1.5 (s, 3H), 1.7 (s, 3H), 4.05 pns isomer A ,. 8S) oil (dd, IH), 4.35 (s, IH), 4.5 (d, IH), 4.65 (m, 2H), 5.25- 5.45 (m, 3H), 5.55 (d, IH), 5.9-6.0 (m, IH), 8.6 (m, 2H),

in

(out

Isomer B, 8S)

Oi

(out

Isomer A, 8S)

( out

Isomer B, 8R)

59

35

oil.

8.85 (s, IH).

1.5 (s, 3H), 1.8 (s, 3H), 4.15 (dd, IH), 4.25 (bs, IH), 4.55 (s, IH), 4.7 (m, 2H), 5.2-6.2 (m, 5H) , 8.67 (bs, 2H), 9.00 (bs, IH).

1.51 (s, 3H), 1.74 (s, 3H), 4.01 (dd, IH), 4.44 (d, IH), 4.60 (s, IH), 4.70 (d, 2H), 5.40 (m, 3H), 5.60 (d, 1H), 5.96 (m, IH), 7.34 (t, IH), 8.80 (d, 2H).

1.52 (s, 3H), 1.78 (s, 3H), 4.23 (m, 2H), 4.64 (s, IH), 4.71 (d, 2H), 5.30-5.46 (m, 3H), 5.53 (d, IH) , 5.90-6.04 (m, IH), 7.34 .Ct, lH), 8.85

, (d, 2H).

Example 58 (continued)

13

Vf

% Yield

100 H-NMR _(CDCl3) ppm (delta);

1.47 (s _f 3H), 1.67 (s, 3H),

4.16 (m, 2H), 4.64 (π, 3Η),

5.47 (m, 4H), 5.94 (m, IH),

7.43 (m, 3H), 8.02 (m, 3H).

(8R)

\ _

/ ~ (no cleaning)

36

(after cleaning)

(8S)

1.42 (s, 3H), 1.7 (s, 3H), (d of d, J = 4 and 8, IH), (s, IH), 4.65 (m, 2H), 5.2-6.3 (m, 3H), 5.6 (d, J = 4, IH), (d, J = 8, IH), 7.2-7.6 (m, 2H), 7.8-8.1 (m, 2H).

1.45 (s, 3H), 1.55 (s, 3H), 4.15 (d of d, IH), 4.5 (s, IH), 4.65 (m, 2H), 5.2-6.4 (m, 5H), 7.2-7.6 ( m, 2H), 7.8-8.2 (m, 2H).

Example 58 (continued)

C. Reaction of the allyl esters prepared above by the method of Example 60 provided the following potassium salts.

OH R ¹³ -CH,

-N-

^CH

''COOK

13

Cl

'N (isomer A)

(Isomer A)

(Isomer B)

% Stereo yield _chemistry

(Isomer B)

8R

8S

8S

8R H-NMR (DMSO-d ₆ ) ppm (delta):

1.46 (s, 3H), 1.56 (s, 3H), 3.76 (s, IH), 4.13 (dd, "IH), 5.05 (d, IH), 5.4 (d, IH), 8.58 (d, 2H), 8.72 (s, IH) Infrared (KBr): 3478, 1761, 1607 cm " ¹ .

1.42 (s, 3H), 1.65 (s, 3H), 4.25 (S, 3H), 4.27 (dd, IH), 5.3 (d, IH), 5.35 (d, IH), 8.6-8.7 (m, 2H) , 8.8 (m, IH).

(D ₂ O): 1.32 (s, 3H), 1.54 (S, 3H), 4.18 (S, IH), 4.14-4.19 (m, IH), 5.19 (d, IH), 5.25 (d, IH), 7.45 (t, IH), 8.75 (d, 2H).

(D ₂ O): 1.44 (Ξ, "3H), 1.58 (s, 3H), 4.12 (s, IH), 4.18 (m, IH), 5.22 (d, IH), 5.49 (d, IH), 7.46 (t> .. IH), 8.75 (d, 2H). "-

Example 58 (continued)

13

6 ^H 5

% Stereo

Yield chemistry

97

8R

85

8 p

91

8R H-NMR (DMSO- _dc ) ppm (delta):

1.47 (s, 3H), 1.58 (s, 3H), 3.79 (s, IH), 4.13 (dd, IH), 5.2 (d, IH), 5.4 (d, IH), 5.95 (bs, IH), 7.45 (t, IH), 7.60 (t, 2H), 8.03 (d, 2H), 8.16 (s, IH).

1.48 (s, 3H), 1.67 (s, 3H), 4.27 (d of d, J = 4 and 10, IH), 4.3 (s, IH), 5.45 (d, J = 4, IH), 5.57 (i.e. , J = IO, IH), 7.45-7.65 (m, 2H), 8.04 (t, J = 8, 2H).

IR (KBr): 3424, 1765, 1746, 1592 cm " ¹ .

1.42 (s, 3H), 1.56 (s, 3H), 4.16 (s, IH), 4.25 (d of d, J = 4 and 8, IH), 5.46 (d, J = 4, IH), 5.5 (i.e. , J = 8, IH), 7.4-7.6 (m, 2H), 7.8-8.05 (m, 2H).

Preparation A e-Chloropyridine ^ -ylmethyltriphenylphosphonium chloride

(i) 6-Chloro-2-methylpyridine-1-oxide

To a solution of 5/1 g (40 mmol) of 6-chloro-2-picoline in 50 ml of methylene chloride was added 8.625 g (40 mmol) of 80% m-chloroperbenzoic acid, and the mixture was stirred at room temperature for 15 h. The reaction was quenched with 0.5 mL of saturated sodium thiosulfate and adjusted to pH 7.5 with sodium bicarbonate solution. The organic layer was washed with water, dried (over Na ₂ SO ₄ ), and solvent evaporated to give 4.84 g of N-oxide. ¹ H-NMR _(CDCl3) ppm (S) '- 2.6 (s, 3H), 7.0-8.0 (m, 3H).

(ii) 6-Chloro-2-acetoxymethylpyridine

A solution of 4.8 g (0.035 mol) of the above N-oxide in 15 mL of acetic anhydride was heated at 100 ° C for 1 h and distilled in vacuo to give 2.39 g of the desired product, b.p. 125-128 ° / o , 7 mm, to give as a colorless oil. ¹ H-NMR _(CDCl3) ppm {€): 2.1 (s, 3H), 5.1 * (s, 2H), 7.0-7.8 (m, 3H).

(iii) 6-chloro-2-pyridylmethanol

Hydrolysis of the product obtained in part (ii) in 10 ml of 2 N hydrochloric acid at 70 ° C for 1 h, followed by neutralization (K ₂ CO ₃ ), extraction with chloroform and evaporation of solvent from the dried extract gave 1.87 g of crude Alcohol, which was purified by column chromatography on silica gel to give 0.982 g of pure material. ¹ H-NMR _(CDCl3) ppm (£): 4.8 (s, 2H), 5.3 (bs, 1H), 7.0-7.8 (m, 3H).

Preparation A (continued) (iv) 6-Chloro-2-chloromethylpyridine

6-Chloro-2-pyridylmethanol, 0.982 g (6.84 mmol) in 10 mL of methylene chloride was treated with 0.814 g of thionyl chloride at room temperature for 1 h. The mixture was neutralized with saturated sodium bicarbonate solution, extracted with methylene chloride, the extracts dried and solvent evaporated to give 815 mg of product as colorless crystals. ¹ H-NMR _(CDCl3) ppm (S): 4.7 (s, 2H), 7.1-8.0 (m, 3H).

(v) The Wittig reagent was prepared by dissolving 815 mg (5 mmol) of the product obtained in part (iv) and 1.318 g (5 mmol) of triphenylphosphine in toluene (10 ml) and heating the mixture to reflux for 6 h. The precipitated product was collected by filtration to give 1.368 g (65%) of the title compound. H-NMR (DMSO) ppm [S] : 5.5 (s, 1H), 5.8 (s, 1H), 7.2-8.2 (m, 18H).

(vi) * 4-methoxypyridin-2-ylmethyltriphenylphosphonium chloride

Starting from 2-methyl-4-methoxypyridine-1-oxide (2.1 g) in the manner of part (ii), 2.5 g of 2-acetoxymethyl-4-methoxypyridine gave about 25% of 5-acetoxy-2 methyl-4-methoxypyridine was contaminated. This mixture was dissolved in methanol (10 ml) containing 1.118 g (20.7 mmol) of sodium methylate and stirred for one hour at reflux. The methanol was evaporated in vacuo, the residue diluted with water, neutralized with dilute hydrochloric acid and extracted with chloroform. The organic layer was washed with brine, dried over MgSO 4, and concentrated in vacuo to give 853 mg (41%) of 2-hydroxymethyl-4-methoxypyridine. ¹ H-NMR (CDCl ₃₎ pp (£): 3.9 (s, 3H), 4.72 (s, 2H), 5.35 (bs, 1H), 6.7 (dd, 1H), 6 , 95 (d, 1H), 8.3 (d, 1H).

The hydroxymethyl compound obtained above was converted into 2-chloromethyl-4-methoxypyridine by the method of the above part (vi) to give 0.895 g (5.68 mmol). This was reacted with an equimolar amount of triphenylphosphine in toluene (10 ml) at reflux for 20 h. The precipitated product was filtered off to give 860 mg of the title compound as a yellow solid.

Preparation B 2-Quinolinylmethyltriphenyl phosphonium chloride

A solution of 2-chloromethylquinoline, 6.26 g (0.035 mol) and 9.20 g (0.035 mol) of triphenylphosphine in 80 ml of toluene was heated at reflux for 3 hours. The precipitated solid was collected on a filter and dried in vacuo to give 3.5 g (23%) of product as a brown solid.

Production C

3 ~ allyloxy-2-pyridylmethyltriphenyl-phosphonium chloride (i) 3-allyloxy-2-hydroxymethylpyridine

To a sodium methylate / methanol mixture of 1.43 g (62 mmol) of sodium metal and 100 ml of methanol was added 5.9 g (31 mmol) of 3-hydroxy-2-hydroxymethylpyridine and the methanol was removed in vacuo. The resulting residue was dissolved in 80 ml of dimethyl sulfoxide (DMSO) and 3.0 ml (34.7 mmol) of allyl bromide in 20 ml of DMSO was added over 20 minutes at room temperature. The mixture was stirred for 2 h, the DMSO distilled in vacuo and the residue partitioned between chloroform / water. The aqueous phase was adjusted to pH 7.5 and extracted three times with chloroform. The combined organic layers were washed with water, brine, dried over Na 2 SO 4

and concentrated to give 3.4-8 g (68%) of the desired ether.

(ii) 3-Allyloxy-2-chloromethylpyridine

The product from Part A (3.43 g, 20.8 mmol) in 20 mL of methylene chloride was treated with 1.5 equivalents (2.5 mL) of thionyl chloride and the mixture stirred under nitrogen for 2 h. The volatiles were removed in vacuo and the residue partitioned between methylene chloride and water. The aqueous layer was adjusted to pH 7.5 and extracted with fresh methylene chloride. The combined extracts were washed with water, brine, dried (over 2SO ^ ₁ J) and the solvent removed in vacuo (86%) of the desired product to yield 3.28 g, which was used in the next step.

(iii) The product from part (ii) (3.28 g, 17.9 mmol) was dissolved in 30 mL of toluene and 4.69 g (17.9 mmol) of triphenylphosphine were added. The mixture was allowed to stand for 3 h Reflux ^ then stirred for 12 h at room temperature. The product was isolated by filtration washing with toluene to provide 3.89 g (49%) of the desired Wittig reagent.

Production D

Allyl 6-wf-bromo-i, 1-dioxopenicillanate (i) 6-o (-Brompenicillanic Acid 1,1-dioxide

A suspension of 20.26 g (0.0517 mol) of 6,6-dibromophenicillanic acid 1,1-dioxide in 80 ml of water was treated in portions with 13 g (0.155 mol) of solid sodium bicarbonate. The vigorous evolution of gas was controlled by the addition of ethyl acetate. Solid sodium bisulfite, 6.76 g (0.062

Mol) was then added in portions, the resulting mixture was stirred for 35 minutes and adjusted to pH 1.0 with concentrated hydrochloric acid. The acidified mixture was diluted with ethyl acetate, the organic phase washed with brine, dried (over Na 2 SO 4) and solvent removed in vacuo. The residue was triturated with chloroform and filtered to give 6.72 g of pale yellow solid. An additional 3.2 g of product was obtained by concentrating the filtrate and treating the residue with chloroform.

(ii) To 6.352 g of the material of the first crop from above in 20 ml of dimethylformamide was added 1.76 ml (20.3 mmol) of allyl bromide, 2.83 ml (20.3 mmol) of triethylamine and 0.2 g of sodium bicarbonate. and the mixture stirred for 15 h at room temperature under nitrogen. Water was added, the mixture extracted with ethyl ether, the extracts washed with water, dried over Na 2 SO, and the solvent removed in vacuo to give 4.60 g (64%) of the desired ester as an oil. ¹ H-NMR _(CDCl3) ppm {£): 1.4 (s, 3H), 1.6 (s, 3H), 4.4 (s, 1H), 4.6 (d, 1H), 4 , 7 (d, 2H), 5.15 (d, 1H), 5.1-5.95 (m, 3H).

(iii) allyl-6,6-dibromophenicillanate

Esterification of the 6,6-dibromophenicillanic acid by the above procedure on a 0.417 molar scale gave 140 g (84%) of allyl ester as an oil. ¹ H-NMR _(CDCl3) ppm (£) / 1,5 (s, 3H), 1.65 (s, 3H) 4.6 (s, 1H), 4.75 (m, 2H), 5 , 3-5.6 (m, 2H), 5.85 (s, 1H), 5.8-6.3 (m, 1H).

Production E

2-Formyl-1-methylimidazole 2-hydroxymethyl-1-methylimidazole

A mixture of 50 g of 1-methylimidazole and 100 ml of 37% formaldehyde (wt% 1.08) was placed in a bomb of rust

I IU

brought free steel (300 ml) and heated to 150 ° (bath temperature) for 17 h. The bomb was then cooled in ice and the mixture removed, concentrated in vacuo and the residue stored at 4 ^° C. overnight. The resulting mixture of crystals and oil was filtered, washing with ethyl acetate. The colorless crystals were dried in vacuo to yield 14.60 g of product. A second crop of 6.48 g was obtained by reprocessing the mother liquor. Total yield 21.08 g (31%).

(ii) To a solution of 4.96 g (43.9 mmol) of 2-hydroxymethyl-1-methylimidazole in 50 ml of dioxane was added 4.90 g (44.1 mmol) of selenium dioxide and the mixture was stirred at 85-90 for 5 h C., stirred at room temperature for 36 h, at 85 ° C. for 8 h and finally at room temperature for 15 h. The reaction mixture was filtered, the solvent removed in vacuo to give 4.81 g of crude aldehyde, which was distilled to give 2.11 g of product as colorless crystals, b.p. 65 ° C at 2.8 mm Hg.

Production F

6-methyl-2-pyridylmethyltriphenylphosphonium chloride

(i) 6-methyl-2-hydroxymethylpyridine

6-methylpyridine-2-carboxaldehyde (0.44 mmol) in 50 ml of methanol was reduced with 20.6 mmol of sodium borohydride at 0 to 5 ⁰ C. After the reduction was complete, the mixture was neutralized with 2N sulfuric acid (pH 7.5), filtered, the filtrate concentrated and partitioned between chloroform and water. Evaporation of the solvent from the organic layer gave 3.32 g of black-red oil, which was used in the next step.

(ii) 2-chloromethyl-6-methylpyridine

The above product, 3.32 g (0.27 mmol) in 20 ml of methylene

Chloride was treated with 1.94 ml (27 mmol) of thionyl chloride at room temperature for 1 h. The mixture was neutralized (with NaHCO 3) and extracted with chloroform. Evaporation of the solvent gave 3.22 g of product as an oil which was used in the next step.

(iii) A solution of 3.22 g of the oil from part (ii), 5.96 g of triphenylphosphine in 30 ml of toluene was heated at reflux for 4 hours. Filtration of the precipitate gave 3.93 g of Wittig reagent as a brown solid.

Production G

2-pyrazinylmethyltriphenylphosphonium chloride (i) 2-hydroxymethylpyrazine

To a solution of 11.29 g (79.2 mmol) of 2-pyrazinecarbonyl chloride (prepared by treating the corresponding 2-carboxylic acid with a molar excess of thionyl chloride at reflux temperature) in 100 ml of tetrahydrofuran at -78 ° C was added over 20 min 2, 0 g, 52.6 mmol, lithium aluminum hydride (95% pure) added in portions. The mixture was stirred for 10 minutes and then allowed to warm to room temperature. The reaction was quenched with 2N sodium hydroxide, filtered, washing with methanol. Concentration of the filtrate in vacuo gave 4.12 g of dark solid, which was used in the next step.

(ii) The above dark solid (4.12 g, 37.8 mmol) was dissolved in methylene chloride and 2.8 mL of thionyl chloride was added at ⁰ ° C. The mixture was warmed to room temperature, stirred for 30 minutes, water added, the mixture neutralized and extracted with methylene chloride to afford 2.29 g of yellow oil, which was used in the next step.

(iii) To the above oil, 2.29 g, in 40 ml of toluene was added 4.70 g of triphenylphosphine and the mixture was refluxed for 3 hours. Workup in the usual manner yielded T, 995 σ Wittig reagent as a brown solid.

Production. H 4-formylpyrimidine

A solution of 4-methylpyrimidine (10 g, 0.106 mol) in 100 ml of dioxane was treated with 11.8 g of selenium dioxide at room temperature and the mixture heated to 100 ° C for 15h. After addition of 2.5 g of selenium dioxide was further heated for 1 h, the mixture cooled, filtered and the cake washed with ethyl acetate. The filtrate and washings were evaporated to dryness in vacuo. The residual dark oil was taken up in methylene chloride, filtered and the solvent was evaporated. The residue was crystallized from a small amount of methylene chloride to afford the title aldehyde. ¹ H-NMR _(CDCl3) ppm (£): 7.87 (dd, 1H), 9.06 (d, 1H), 9.43 (d, 1H), 10.0 (s, 1H).

Production I Diphenylmethyl-6-oi-bromo-1,1-dioxopenicillanate

To a solution of 21.557 g (0.1 mol) of diphenyldiazomethane in 400 ml of dry tetrahydrofuran were added over 30 min 31.2 g (0.1 mol) of 6-oC-bromo-i, 1-dioxopenicillansäure in portions. The reaction was mildly exothermic. The mixture was stirred for 1 h, the solvent was evaporated and the residue was dissolved in ethyl acetate (50 ml). Ethyl ether (400 ml) was added and the resulting mixture stored at 4 ⁰ C. After 18 h, no crystals formed. The mixture was then concentrated in vacuo to give 51.2 g of yellow solid.

To provide material which was chromatographed on silica gel eluting with chloroform to afford 14.86 g of colorless product as a glass. ¹ H-NMR _(CDCl3) ppm (S): 1.26 (s, 3H), 1.57 (s, 3H), 4.55 (s, 1H), 4.70 (d, 1H), 5 , 13 (d, 1H), 6.9 (s, 1H), 7.27 (s, 10H).

° - π ι ft Β - 31643

Preparation J Benzothlazo-2-carboxaldehyde

A solution of freshly distilled (bp. 82 ° C / 2 torr) benzothiazole (10 g, 0.074 mol) in 250 ml tetrahydrofuran was cooled under nitrogen to -78 ⁰ C and stirred at this temperature for 15 min. To this was added dropwise over 15 minutes 29.6 ml (0.074 mol) of 2.5 M n-butyllithium, stirred for a further 25 min, and 8.6 ml (0.111 mol) of dimethylformamide were added and the reaction mixture was stirred for 1 h. The mixture was allowed to warm to room temperature, 200 g of ice were added and the solvent removed in vacuo, the aqueous residue was extracted with 5x100 mL of ethyl acetate, the extracts dried over MgSO ₄ , concentrated to a small volume and placed on a silica gel column eluting with 96: 4 chromatographed chloroform / ethyl acetate to give 8.4 g of the desired aldehyde, m.p. 74.5 to 75 ⁰ C. ¹ H-NMR _(CDCl3) ppm (S).:. 7.54 (m, 2H), 7.97 (m, 1H), 8.21 (m, 1H), 10.1 (s, 1H).

The following aldehydes were prepared similarly to the above procedure: Starting compound aldehyde% Yield H-NMR (CDCl - ) ppm (delta):

CH, CH ₁ CH ₁ CH ₇ 98

^J W r = \ yellow-brown oil

: l \ sdp. 120-122 ° C

at 25 torr

CHO

Production J (continued)

Starting compound aldehyde _% Yield H-NMR (CDCl ₃ ) ppm (delta);

N; -N N7-N 50 4.2 (s, 3H), 8.05 (s, IH),

^ N. _cho 10.00 (s, IH).

** , ν 82 1.2 (t, 6H), 3.6 (q, 2H),

(OC ₂ H ₅ J ₂ N _n NCH (OC ₂ H _{5) 2} 3.64 (q, 2H), 6.95 (s, IH),

CHO

7.21 (d, IH), 7.55 (d, IH),

8.23 (d, IH).

Prepared by reacting 1H-1,2,4-triazine with methyl iodide in ethanolic sodium nitrate in 37% yield as a solid.

** Prepared from imidazole and ethyl orthoformate by the method of Curtis et al., J. Org. Chem.

45, 4038 (1980).

-14b-

Production K Pyrazine-2-carboxaldehyde

Pyrazine-2-carboxylic acid was esterified with methanolic sulfuric acid for 5 h at reflux. The methanol was removed in vacuo / diluted with methylene chloride and neutralized with sodium bicarbonate (pH 7/5). The dried organic layer was concentrated / to afford methyl pyrazine-2-carboxylate as an off-white solid in 84% yield, which was recrystallized from isopropanol / ethyl ether to give yellow needles. ¹ H-NMR _(CDCl3) ppm U): 4.1 (s, 3H), 8.93 (m, 2H), 9.74 (m, 1H).

The methyl ester, 20 g (0.145 mol), was dissolved in 600 ml of dry tetrahydrofuran, cooled to -78 ° C, and a solution of 5.8 g of 98% lithium aluminum hydride in THF was added dropwise over 15 min. The mixture was stirred at -78 ° C for 30 minutes, 20 ml of acetic acid was added, and the mixture was concentrated under reduced pressure. The residue was partitioned between 2N hydrochloric acid (30 ml) and chloroform, the combined organic layers were washed with water, dried and evaporated to afford 8.53 g of crude product, which was purified by passing it through a silica gel column Elute with 4: 1 methylene chloride / ethyl acetate to give 5.02 g of light yellow needles. H-NMR _(CDCl3) ppm (6): 8.38 (m, 2H), 8.7 (bs, 1H), 9.7 (s, 1H).

Production L

2-phenyl-1,2,3-triazole-4-carboxaldehyde

(i) reacting acetonedicarboxylic 0.34 mol in 100 ml water with 0.58 mol of sodium nitrite at 0-1O ⁰ C followed by addition of dilute nitric acid to precipitate the product gave a 46% yield of the dioxime, 1,3-

Dioximino-2-oxopropane, as yellow-brown crystals.

(ii) The dioxime (0.158 mol) in ethanol (170 ml) was reacted with equimolar amounts of phenylhydrazine hydrochloride and sodium acetate at 70 ° C for 30 min. Water (170 mL) was added and the mixture warmed to 85 ° C, reduced in volume to 250 mL, and cooled to give 24.7 g of phenylhydrazone. ¹ H-NMR (CD ₃ COCD ₃ ) PPm {£) : 7.3 (m, 5H), 7.9 (s, 1H), 8.6 (s, 1H), 10.5 (bs, 1H) , 11.4 (bs, 1H), 12.3 (s, 1H).

(iii) The above dioximphenylhydrazone, 24.7 g (0.119 mol), was stirred at room temperature with 500 ml of acetic anhydride for 30 min, and the mixture was poured into water, stirred for 20 min, and filtered. The resulting crude solid was recrystallized from benzene / ethyl acetate to afford 15.47 g (52%) of the monoacetate, HON = CH-Ci = NNHC ₆ H ₅ ) CH = NOCOCH ₃ , as a yellow powder. ¹ H-NMR _(CDCl3, acetone): 1.85 (s, 3H), 7.09 (m, 5H), 7.95 (s, 1H), 8.3 (s, 1H), 10.9 (bs, 1H), 12.25 (s, 1H).

(iv) A solution of 15.40 g (0.062 mol) of the monoacetate obtained above and 22.16 g (0.068 mol of cesium carbonate in 400 mL of tetrahydrofuran) was stirred at room temperature for 1 h, filtered and the filtrate concentrated to give a yellow The residue was dissolved in 700 ml of hot 1: 2 isopropyl ether / cyclohexane and concentrated to 200 ml to give 9.41 g (80%) of 2-phenyl-1,2,3-triazole-4-carboxaldehyde oxime as a yellow To supply powder, which was used in the next stage.

(v) A mixture of the oxime, 9.41 g (0.0497 mole) of 4.48 g of s-trioxane and 300 ml of 2N hydrochloric acid was heated at reflux for 3.5 hours. The aqueous phase was extracted with ethyl ether, the combined ether layers washed with water, dried (over MgSO 4), and the ether stripped in vacuo to afford 6 g of crystalline aldehyde. H NMR (CDCl ₃₎ ppm (^). 7.6 (m, 3H), 8.25 (m, 3H), 10.25 (s, 1H), mp 66-67 ° C.

Production M /

S-methylisoxazole-S-carboxaldehyde

(i) Ethyl 5-methylisoxazole-3-carboxylate

A mixture of 0.16 mol of ethyl 2,4-dioxovalerate and O, 08 mol of hydroxylamine sulfate, 50 ml of ethanol and 70 ml of toluene was stirred at 40 ° C for 4 h. The mixture was cooled to 15 to 20 ⁰ C, 1.8 g of concentrated ammonium hydroxide was added and stirring continued for 60 h at room temperature. The mixture was poured into water / toluene, the aqueous layer extracted with toluene, the organic layers combined, washed with brine, and dried (over Na 2 SO 4). Evaporation of the solvent in vacuo gave a yellow liquid which was distilled to afford 14.68 g of product, b.p. 12O-124 ° C / 3O torr, which crystallized on standing.

(ii) The ethyl ester obtained above, 2.0 g (14.4 mmol) in 75 ml of toluene was reduced with an equimolar amount of 1 M diisobutyaluminum hydride in hexane under nitrogen at -75 to -70 ° C. After stirring for 30 minutes, the reaction was quenched with ammonium chloride solution, warmed to room temperature, and the solvent removed in vacuo. The residue was triturated with hot methanol, evaporated to afford 0.85 g of product as a colorless oil. ¹ H-NMR _(CDCl3) ppm (<f): 2.4 (s, 3H), 6.3 (d, 1H), 9.0 (s, 1H).

2υ. 1.83- 3it482

Claims (6)

invention claim 1 8
Formula (II ¹ ) / wherein R is acetyl, is deacetylated. -A - 1 ft 1 ft of the formula R Cl or R Br or a corresponding acid anhydride in the presence of an equimolar amount of tertiary
Amines and reacted in the presence of a reaction-inert organic solvent, and when compounds of formula (VIII) are desired, the first product is dehydrated or a compound of 1. A method for producing a compound of the invention O O (Viii) (H ¹ I wherein RR is ^a or R wherein R ^{a is} the residue of a carboxy-protecting group selected from tetrahydropyranyl, allyl, benzyl, 4-nitrobenzyl, benzhydryl, 2,2,2-trichloroethyl / t-butyl and phenacyl, and R is hydrogen or the radical an in vivo readily hydrolysable ester group selected from 3-phthalidyl, 4-crotonolactonyl, γ- butyrolactone-4-yl, -CH ₀ -C = CR ^{4 2} I Il -COCOR ⁶ Λ5 -COCOOR ⁶
15 R and -CHOCOR ¹⁴ a; 's2 2. The method according to item 1, characterized in that the reaction irfit aldehyde, R ¹² R ¹³ CO, at -78 ° C in the presence of tetrahydrofuran or ethyl ether is performed. 2 3
one of the radicals R and R is hydrogen and the other is Cl, CH ₉ OH, vinyl, (C ₁ -C.) alkylthio, (C ₁ -C.) alkylsulfonyl, furyl, thienyl, N-methylpyrrolyl, N-acetylpyrrolyl, R ⁷ C _C H, R ⁷ C _C HS, -CH (R ⁴ JWR ¹⁶ R ¹⁷ , -CWR ⁸ R ⁹ , -CNR ⁸ R ⁹ ,.: _ Ο 4 b 4 HH S NH (R , 11 \ _R ii 11 11 -N, N ll -JJ- 'i <J I _# (R and m is 2 or 3, ρ is zero or 1, t is zero, 1 or 2, X _{1 is} S, O or NR ¹¹ , R ^{7 is} hydrogen, (C ₁ -C ₄ ) Al] CyI, (C ₁ -C ₄ ) alkoxy, allyloxy, hydroxyl, carboxyl, (C ₂ -C ₅₎ alkoxycarbonyl, (C ₁ ~ C ₅₎ alkylcarbonyl _/ phenyl, benzyl, naphthyl, pyridyl, NR ⁸ R ^9, CONR ⁸ R ^9, NHCOR ^10, NO ₀ , Cl, Br, CF-, or SR ⁸ , 8 9 ^{Δ J} R and R are each hydrogen, (C ₁ -C ₄ ) alkyl, phenyl or benzyl, R is (C ₁ -C ₄ ) alkyl, CF ₃ or phenyl and R is hydrogen, CH ₃ , C ₃ H ₅ or CH ₃ CO is, R ¹⁶ and R ¹⁷ respectively H, (C ₁ -C ₄ ) alkyl, Hydroxyalkyl or together and taken with the nitrogen atom to which they are attached, R R is a pyrrolidino, piperidino, morpholino, thiomorpho form lino or 4-methylpiperazino group, and -10 - RH, (C ₂ -C ₅ ) alkanoyl, (C ₂ -C ₅ ) - Alkoxycarbonyl, pyrazinecarbonyl, benzoyl, CF ₃ CO or CONR ⁸ R ⁹ , one of the groups R and R is hydrogen and the other vinyl, (C ₁ -C ₄ ) alkylsulfonyl, furyl, thienyl, N-methylpyrrolyl, N-acetylpyrrolyl, CH (R ⁴ ) NR ¹⁶ R ¹⁷ , CNR ⁸ R ⁹ , CNR ⁸ R ⁹ , Il It , S NH R ' , R N (0) _p ' II - < ^R > t-f 11 .-1II N, V, (r " ^χ ί αϊν or (H ⁷ ) 12 provided that when R or R and ρ is zero, R has a different meaning as H or CH ^, and m is 2 or 3, ρ is zero or 1, 117 t is zero, 1 or 2, X _{1 is} S, O or NR, R is hydrogen, (C ₁ -C-) alkyl, (C ₁ -C.) Alkoxy, allyloxy, hydroxy, carboxyl, (C ₂ ^- C ₅₎ alkoxycarbonyl, (C.-C ₅₎ alkylcarbonyl, phenyl, benzyl, Naphthyl, pyridyl, NR ⁸ R ⁹ , CONR ⁸ R ⁹ , NHCOR ¹⁰ , NO ₂ , Cl, Br, CF ₃ 8 8 9 or SR is and R and R are each H, (C ₁ -C ₄ ) alkyl, phenyl or benzyl, R is (C ₁ -C ₄ ) alkyl, CF, or phenyl, and 11 1J R is H, CH ₃ , C ₃ H ₅ or CH ₃ CO, R and R are each H, (C ₁ -C ₄ ) alkyl, (C ₂ -C ₄ ) hydroxyalkyl
are or together form the nitrogen atom to which they are attached, R and R form a pyrrolidino, piperidino, morpholino, thiomorpholino or 4-methylpiperazino group, or a pharmaceutically acceptable one
Salt of the compound,
characterized in that a compound of the formula 0 0 (Ix) wherein R is other than H, with an equimolar amount of a (C .. -C.) Alkylmagnesiumhalogenids in an ethereal solvent at a temperature of -80 ° to 25 ° C to a first product of the formula OH 0 0 IT '\ / rw 1X1 Cf -N L. is reacted, and when compounds of the formula (II ¹ ), wherein R has a meaning other than H, the first product with an acyl chloride, acyl bromide are desired - 2 \ - AS wherein R and R are each hydrogen or CH, R '14 is and R 15 wherein R is 2-phenylacetamido, 2-phenoxyacetamido, D-2-amino-2-phenylacetamido, D-2-amino-2- (4-hydroxyphenyl) acetamido, 2-carboxy-2-phenylacetamido, 2-carboxy-2 - (2-thienyl) acetamido, 2-carboxy-2- (3-thienyl) acetamido, D-2- (4-ethyl-2,3-dioxopiperazinocarbonylamino) -2-phenylacetamido or 2,2-dimethyl-4 is phenyl-5-imidazolidinon-1-yl, 3. The method according to item 2, characterized in that 12 13
one of the groups R and RH and the other 2-thiazolyl, 3-isothiazolyl, 5-methyl-3-oxazolyl, 1,3,4-thiadiazol-2-yl, pyridazin-3-yl, 1 , 2 , 3-thiadiazine -4-yl, 1,2,4-0xadiazin-3-yl, 1, 2,4-oxadiazine-5-yl, 1,3-oxadiazine-2 _# 4-yl, 1-phenyl-5-methyl-1 , 2,3-triazin-4-yl, N-methyl-1,2,3,4-tetrazino-5-yl, 1-methyl-1,2-diazin-3-yl, 4-pyrimidinyl, 4,5 Dimethylthiazol-2-yl, benzothiazol-2-yl or 1-benzylbenzimidazol-2-yl. 4. The method according to item 3, characterized in that a compound of formula (X) wherein R is allyl or benzyl, with methylmagnesium bromide and an aldehyde 12 13 of the formula RR CO, and a product of formula (II) wherein X is H is obtained by hydrogenolysis with tri-n-butyltin hydride in benzene or toluene as solvent at a temperature of from ⁰ ° C to the boiling point of the solvent. 5. Method according to point 3, characterized in that- 12 1 the groups R and R or benzothiazol-2-yl. 12 13 one of the groups R and R H and the other 2-thiazolyl A process according to any of the preceding points, characterized in that in the starting compound R is allyl and the allyl group is converted to a corresponding compound wherein R is H, Na or K by reacting with catalytic amounts of tetrakis in the presence of a reaction-inert solvent ( triphenylphosphine) palladium (O) and triphenylphosphine and an equimolar amount of the sodium or potassium salt of 2-ethylhexanoic acid allyl ester product obtained.