KR860001289B1 - Process for preparing b-lactam derivatives - Google Patents

Abstract

β-Lactams I [R1=acyl; R2=H, C1-4 alkoxy; R3, R4=H, alkyl, cycloalkyl, Ph, substituted Ph, alkenyl, etc.! were prepd. Thus, a mixt. contg. 100 ml dimethylformamide, (s)-3-amino-2-oxo-1- azetidinsulfonic acid in 2 ml propylene oxide, and 1.5 g tetrabutyl ammonium salt was cooled to 0≰C. This soln. was treated with 0-formyl mandelic chloride in 10 ml acetonitrile and evaptd. The resulting ext. was reacted with 2 g potassium sulfate perfluorobutane in 15 ml acetone and mixed with 200 ml ether to give 1.5 g [3S(±)!-3- [[(formyloxy)phenyl-acetyl!amino!-2-oxo-1-azetidine sulfonic acid potassium salt.

Description

Method for preparing β-lactam antibiotics

The present invention relates to a process for the preparation of novel β-lactam antibiotics.

It has been known that β-lactam nuclei can be biologically activated by sulfonate substituents attached to nitrogen atoms of β-lactam nuclei.

Β-lactams having sulfonate substituents including intramolecular salts in the 1-position and acylamino substituents in the 3-position show effects on Gram-negative and Gram-positive bacteria.

Among the novel β-lactam antibiotics of the invention, compounds of formula (I) below are more suitable.

In addition to the above-described β-lactam having a sulfonate substituent in the 1-position and an acylamino substituent in the 3-position, the present invention has a sulfonate substituent including an intramolecular salt in the 1-position and β- having an amino substituent in the 3-position. It also includes lactams.

Suitable compounds of this type have the general formula (Ia).

These compounds are useful intermediates for the preparation of the corresponding 3-acylamino compounds.

The definitions of symbols used in general formulas (I) and (Ia) and throughout the specification are as follows.

R ₁ is acyl,

R ₂ is hydrogen or alkoxy of 1 to 4 carbon atoms,

R ₃ and R ₄ are the same as or different from each other, and are each hydrogen, alkyl, cycloalkyl, phenyl or substituted phenyl, or either R ₃ or R ₄ is hydrogen, and the other is alkoxycarbonyl, alkene-1- 1, alkyn-1-yl, 2-phenylethenyl or 2-phenylethynyl,

는 수소 또는 양이온이다.M

Is hydrogen or cation.

"Alkyl" and "alkoxy" are straight and branched groups, groups having 1 to 10 carbon atoms are suitable.

"Cycloalkyl" and "cycloalkenyl" are 3, 4, 5, 6 or 7 cycloalkyl and cycloalkenyl groups.

"Alkenyl" is a straight and branched group, with 2 to 10 carbon atoms being suitable.

"Halogen" is fluorine, chlorine, bromine and iodine.

"Substituted phenyl" is a phenyl group substituted with one, two or three amino, halogen, hydroxyl, trifluoromethyl, lower alkyl or lower alkoxy groups.

"Protected carboxyl" is a carboxyl group esterified with a conventional ester protecting group. Such groups are known in the technical art of this field (see US Pat. No. 4,144,333, filed March 13, 1979). Suitable ones for protected carboxyl groups are benzyl, benzhydryl and t-butyl ester.

“Acyl” includes various organic radicals derived by the removal of hydroxyl groups from organic acids (ie carboxylic acids). Certain acyl groups are suitable, of course, but they should not be taken as limiting the scope of the invention. Representative acyl groups are acyl groups that have been used in the past for acylating β-lactam antibiotics, including 6-aminopheniclanic acid and derivatives thereof and 7-aminocephalosporanic acid and derivatives thereof. [Cephalosporins and Penicillins, published by Flynn, Academic Press in 1972, Federal Republic of Germany Publication No. 2716677, published October 10, 1978, Belgium, issued December 11, 1978. Patent No. 867994, United States Patent No. 4,422, May 1, 1979, United States Patent No. 3971778, July 27, 1976, United States Patent No. 472199, October 23, 1979, and March 27, 1974. British Patent No. 1348894]. The above documents describing various acyl groups are for reference only. The following acyl groups are intended to specifically illustrate the term "acyl" and are not intended to limit the scope of this term.

As a typical acyl group

의 지방족기(a) general formula

Aliphatic group

Wherein R ₆ is alkyl, cycloalkyl, alkoxy, alkenyl, cycloalkenyl, cyclohexadienyl, or substituted with one or more halogen, cyano, nitro, amino, mercapto, alkylthio or cyanomethylthio groups Alkyl or alkenyl)

(b) a carbocyclic aromatic group having the following general formula

(Wherein n is 0, 1, 2 or 3, and R ₆ , R ₇ and R ₈ are different, respectively, hydrogen, halogen, hydroxyl, nitro, amino, cyano, trifluoromethyl, carbon atom 1 ˜4 alkyl, alkoxy or aminomethyl of 1 to 4 carbon atoms,

R ₉ is amino hydroxyl, carboxyl salt, protected carboxyl, formyloxy, sulfo salt, sulfoamino salt, azido, halogen, hydrazino, alkylhydrazino, phenylhydrazino or [(alkylthio Thioxomethyl] thio.

Carbocyclic aromatic acyl groups having the following general formula are suitable.

(R₉은 카르복실염 또는 술포염이 적합하다)

(R ₉ is preferably carboxyl or sulfo salt)

(R₉은 카르복실염 또는 술포염이 적합하다)

(R ₉ is preferably carboxyl or sulfo salt)

(c) heteroaromatic groups of the general formula

(In the above formula,

n is 0, 1, 2 or 3

R ₉ is as defined above,

R ₁₀ is a substituted or unsubstituted 5-, 6- or 7-membered heterocyclic ring containing 1, 2, 3 or 4 (where 1 or 2 is suitable) nitrogen, oxygen and sulfur atoms)

Representative heterocyclic rings are thienyl, furyl, pyrrolyl, pyridinyl, pyrazinyl, thiazolyl, morpholinyl, pyrimidinyl and tetrazolyl. Representative substituents are halogen, hydroxyl, nitro, amino, cyano, trifluoromethyl, alkyl of 1 to 4 carbon atoms or alkoxy of 1 to 4 carbon atoms.

Wherein R ₁₀ is 2-amino-4-thiazolyl, 2-amino-5-halo-4-thiazolyl, 4-aminopyrimidin-2-yl, 5-amino-1, 2, 4-thiadia Heteroaromatic acyl groups that are sol-5-yl, 2-thienyl or 2-furanyl are suitable.

(d) [[(4-substituted-2, 3-dioxo-1-piperazinyl) carbonyl] amino] arylacetyl group of the following general formula

(In the above formula,

과 같은 탄소환식방향족 및 R₁₀의 정의에서 언급한 이종방향족을 비롯한 방향족기이고,R ₁₁ is a structural formula

Aromatic groups including carbocyclic aromatics such as and heteroaromatics mentioned in the definition of R ₁₀ ,

여기에서 R11은 위에서 정의한 바와 같다) 또는 알킬카르보닐 아미노이다)R ₁₂ is alkyl, substituted alkyl, wherein the alkyl group is substituted with one or more halogen, cyano, nitro, amino or mercapto groups, arylmethyleneamino (ie, -N = CH-R ₁₁ , wherein R ₁₁ is as defined above, arylcarbonylamino (ie

Where R 11 is as defined above) or alkylcarbonyl amino)

[[4-substituted-2, 3-dioxo-1-piperazinyl) carbonyl] amino] Suitable among the arylacetyl groups are those wherein R ₁₂ is ethyl phenylmethyleneamino or 2-furylmethyleneamino.

(e) (substituted oxyamino) arylacetyl groups of the general formula

(In the above formula,

R ₁₁ is as defined above,

여기에서 R₁₁은 위에서 정의한 바와 같다)이거나, 치환된 알킬(여기에서, 알킬기는 1이상의 할로겐, 시아노, 니트로, 아미노, 메르캅토, 알킬티오, R₁₁에서 정의된 방향족기, 카르복실(그 염도 포함), 아미도, 알콕시카르보닐, 페닐메톡시카르보닐, 디페닐메톡시 카르보닐, 히드록시알콕시포스피닐, 디히드록시포스피닐, 히드록시(페닐메톡시) 포스피닐, 또는 디알콕시 포스피닐 치환체로 치환됨)이다.R ₁₃ is hydrogen, alkyl, cycloalkyl, alkylaminocarbonyl, arylaminocarbonyl (i.e.

Wherein R ₁₁ is as defined above or substituted alkyl, wherein the alkyl group is at least one halogen, cyano, nitro, amino, mercapto, alkylthio, aromatic group as defined in R ₁₁ , carboxyl Salts), amido, alkoxycarbonyl, phenylmethoxycarbonyl, diphenylmethoxy carbonyl, hydroxyalkoxyphosphinyl, dihydroxyphosphinyl, hydroxy (phenylmethoxy) phosphinyl, or dialkoxy phosph Substituted with a finyl substituent).

Suitable among the (substituted oxyimino) aryl acetyl groups are groups wherein R ₁₁ is 2-amino-4-thiazolyl. Also suitable are groups in which R ₁₃ is methyl, ethyl, carboxymethyl or 2-carboxyisopropyl.

(f) (acylamino) arylacetyl group of the following general formula

(In the above formula,

R ₁₁ is as defined above,

아미노, 알킬아미노, (시아노알킬) 아미노, 아미도, 알킬아미도, (시아노알킬)아미도,R ₁₄ is

Amino, alkylamino, (cyanoalkyl) amino, amido, alkylamido, (cyanoalkyl) amido,

Suitable as the (acylamino) arylacetyl group of the general formula are those wherein R ₁₄ is amino or amido. Also suitable are groups in which R ₁₁ is phenyl or 2-thienyl.

(g) [[[3-substituted-2-oxo-1-imidazole ridinyl] carbonyl] amino] arylacetyl group of the following general formula

(In the above general formula

(여기에서, R₁₆은 수소,알킬 또는 할로겐으로 치환된 알킬이다), R₁₁에서 상기 정의한 바와 같은 방향족기, 알킬 또는 치환된 알킬(여기에서, 알킬기는 1 이상의 할로겐, 시아노, 니트로, 아미노 또는 메르캅토기로 치환됨)이다.R ₁₁ is as defined above, R ₁₅ is hydrogen, alkylsulfonyl, arylmethyleneamino (ie, -N = CH-R ₁₁ , where R ₁₁ is as defined above),

(Wherein R ₁₆ is hydrogen, alkyl or alkyl substituted with halogen), an aromatic group, alkyl or substituted alkyl as defined above in R ₁₁ , wherein the alkyl group is at least one halogen, cyano, nitro, amino Or a mercapto group).

Suitable among the [[[3-substituted-2-oxo-1-imidazolidinyl] carbonyl] amino] arylacetyl groups of the above general formula are those wherein R ₁₁ is phenyl or 2-thienyl.

Also suitable are groups in which R ₁₅ is hydrogen, methylsulfonyl, phenylmethyleneamino or 2-furylmethyleneamino.

₃M

"치환체는 모든 술폰산염을 포함한다. 물론, 기타염들도 본 발명의 최종 생성물을 정제하거나 약학적으로 허용되는 염을 제조하는데 중간체로서 유용하지만, 약학적으로 허용되는 염이 적합하다. 본 발명의 술폰산염의 양이온 부분은 유기나 무기염기로부터 얻을 수 있다. 이러한 양이온 부분으로는 암모늄, 예를들어 테트라-n-부틸암모늄(이후부터는 테트라부틸암모늄이라 함)같은 알킬암모늄처럼 치환된 암모늄, 리튬, 나트륨 및 칼륨 같은 알카리 금속, 칼슘 및 마그네슘과 같은 알카리 토금속, 피리디늄, 디시클토헥실암모늄, 히드라바미늄, 벤자티늄, N-메틸-D-글루카미늄 이온 등이 있으며, 이 양이온들은 위에 예시한 양이온들로 한정되지 않는다.The term "cationic" as used throughout the specification is any positively charged atom or group of atoms. "-SO of the nitrogen atom in the β-lactam of the present invention

₃ M

"Substituents include all sulfonates. Of course, other salts are also useful as intermediates to purify the final product of the present invention or to prepare pharmaceutically acceptable salts, but pharmaceutically acceptable salts are suitable. The cationic portion of the sulfonate of can be obtained from organic or inorganic bases, such as ammonium, for example substituted ammonium, lithium, such as alkylammonium such as tetra-n-butylammonium (hereinafter called tetrabutylammonium). Alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium, pyridinium, dicyclotohexylammonium, hydravaminium, benzatinium, N-methyl-D-glucanium ions, and the like cations are shown above. It is not limited to one cation.

는 수소일 수 있다.As described in general formula (I) and its definition, M

May be hydrogen.

As described below, the β-lactam of the present invention can be produced by a synthesis method. The nonalkoxylated 4-unsubstituted β-lactams of formula (I), ie compounds of formula (I) wherein R ₂ , R ₃ and R ₄ are hydrogen, are 6-amino-phenylsilane acids or 6 as starting materials. It may be prepared using acylaminophenic silane acid. Β-lactams of formula (I) wherein R ₂ is alkoxy can be prepared from the corresponding nonalkoxylated β-lactams. Some of the compounds of the present invention can be crystallized or recrystallized from a solvent comprising water. In such cases, hydrates may be produced. The present invention also encompasses compounds containing variable amounts of water that can be produced by processes such as stoichiometric hydrate and lyophilization.

Some of the β-lactams of formula (I) have been prepared by biological methods. Strains of microorganism Chromobacterium violaceum SC 11, 378 to produce salts of (R) -3- (acetyl amino) -3-methoxy-2-oxo-1-azetidinesulfonic acid It was. For example, various acetic acid bacteria such as Gluconobacter SC11, 435 were cultured to obtain (R) -3-[[N- (Dr-glutamyl) -D-alanyl] amino] -3-methoxy- A salt of 2-oxo-1-azetidinesulfonic acid was produced.

Β-lactams having sulfonate substituents at the 1-position and amino or acylamino substituents at the 3-position have at least one chiral center-carbon atom at the 3-position of the β-lactam nucleus, wherein the amino or acylamino substituents are Are combined. The present invention provides the above-described β-lactam, wherein β-lactam is produced, wherein the three-dimensional structure at the chiral center at the 3-position of the β-lactam nucleus is a natural penicillin (eg, penicillin G). Is the same as the stereo configuration of the carbon atom at the 6-position of and the stereo configuration of the carbon atom at the 7-position of natural sepamycin (eg, sepamycin C).

Suitable among the β-lactams of the general formulas (I) and (Ia) are those having a stereostructure at the chiral center in the 3-position. According to the nomenclature, the compound of formula (I) and (Ia) R ₂ is hydrogen are the compounds of formula (I) and (Ia) R ₂ is alkoxy have the S stereo arrangement has the R stereo arrangement.

Racemic mixtures comprising the above β-lactams are also included in the scope of the present invention.

β-lactams having sulfonate substituents at the 1-position of β-lactam nuclei and amino or acylamino substituents at the 3-position are effective against gram negative and positive groups. Sulfonate substituents are important for the potency of the compounds of the present invention. Compounds in which R ₃ and / or R ₄ are hydrogen or alkyl (particularly methyl) show particularly useful effects.

The compounds of the present invention can be used to treat bacterial infectious diseases including urinary tract infections and respiratory infections in mammals such as domestic animals (eg dog cats, cattle, horses, etc.) and humans.

To treat bacterial transmission of a mammalian animal, a compound of the present invention is administered at a dosage of about 1.4 mg / kg / day to about 350 mg / kg / day, suitably about 14 mg / kg / day to about 100 mg / kg / day. Can be administered to mammals. Dosing methods conventionally used to deliver penicillin and cephalosporin to the infectious site can also be used in the administration of the novel β-lactams of the invention. Such dosing methods include oral, intravenous, intramuscular injection and suppository medication.

The β-lactam product of the present invention is generally prepared by introducing a sulfonic acid substituent (sulfone group-SO ₃ −) to a nitrogen atom at the 1-position of the β-lactam nucleus. This sulfonation reaction is carried out by treating β-lactam with a sulfur trioxide complex or with an equivalent sulfonating agent such as chlorosulfonate.

The most commonly used sulfur trioxide complexes are pyridine-sulfur trioxide, lutidine-sulfur trioxide, dimethylformamide-sulfur trioxide and picoline-sulfur trioxide. Instead of using a previously prepared complex, this complex can be prepared using chlorosulfonyl-trimethylsilyl ester and pyridine as original reagents. Alternatively, the sulfonation reaction can be carried out by compound, for example by first silylating the nitrogen atom of β-lactam nucleus and then silylating the silylated compound with trimethylsilylchlorosulfonate or similar reagents. The reaction can be performed to sulfonate. Representative silylating agents are monosilyl trifluorotoacetamide, trimethylsilylchloride / triethylamine and bis-trimethylsilyl trifluorotoacetamide.

In general, the sulfonation reaction is carried out in the presence of an organic solvent such as pyridine or a mixture of organic solvents (a mixture of a polar solvent such as dimethylformamide and a halogenated hydrocarbon such as dichloromethane is suitable).

The compound produced initially in the sulfonation reaction is a salt of sulfonated β-lactam. When using pyridine-sulfur trioxide as the sulfonated complex, the initially produced compound is the β-lactam-sulfonated pyridinium salt of sulfonated β-lactam.

(Wherein M ⁺ is a pyridinium ion)

These complexes can be converted to other sulfonates using conventional methods such as ion exchange resins, crystallization or ion pair extraction. These conversion methods are also useful for the process of purifying the product. In particular, pyridine salts are converted to potassium salts using potassium phosphate or potassium potassium hexanoate, tetrabutylammonium salts using tetrabutyl ammonium hydrogen sulphate, or positive with M ⁺ being hydrogen using formic acid. It is advantageous to convert to ions.

The sulfonation reaction in which a sulfo group is introduced into the nitrogen atom of the beta -lactam nucleus can be performed by a multistage synthesis step performed before forming the beta -lactam nucleus as described below. The sulfonation reaction is usually carried out at room temperature in the presence of the solvent described above. When there is an amino group, it is suitable to protect an amino group.

As an example, the sulfonation reaction using a benzyloxycarbonyl protecting group can be represented as follows.

Other protecting groups can be used to protect the amine groups, for example with t-butyloxycarbonyl groups, single acyl groups such as acetyl or benzoyl of phenyl acetyl, triphenylmethyl groups or amino groups in azido form. Next, the desired acyl group R ₁ can be introduced by a conventional acylation reaction.

Exemplary acylation methods for converting a compound of formula (III) to a compound of formula (I) include reaction with a carboxylic acid (R ₁ -OH) or a corresponding carboxylic acid halide or carboxylic anhydride. . When R <_2> is alkoxy, acylation is best performed using an acid chloride or an acid bromide.

The reaction with the carboxylic acid can be most easily done in the presence of a carbodiimide such as dicyclohexyl carbodiimide and a substance capable of forming an active ester (eg N-hydroxybenzotriazole). If the acyl group (R ₁ ) comprises reactors such as amino or carboxyl groups, these reactors must first be protected first, then acylation is carried out and finally the protecting groups have to be removed from the resulting material. Alternatively, the sulfonation reaction can be carried out with an acyl group in place. In other words,

If R ₂ is lower alkoxy, additionally, conventional methods are used in which the R ₂ -lower alkoxy group is sulfonated and then introduced or the acylated nitrogen atom in the 3-position is chlorinated and then reacted with the lower alkoxide. Can be introduced prior to the sulfonation reaction.

Acyl groups in the reaction are groups that can be easily removed after the reaction to function as protecting groups and to produce a "deacylated" product (-NH ₂ ).

In addition, the β-lactam ring may be formed by a cyclization reaction, and the sulfonation reaction may be performed before or after the cyclization reaction. In other words,

The acyl group in this reaction is a group that functions as a protecting group and can be easily removed after producing the -NH ₂ product.

An azetidinone starting material wherein R ₂ is hydrogen and at least one of R ₃ and R ₄ is hydrogen can be prepared from amino acids of the general formula (XII).

(At least one of R ₃ and R ₄ is hydrogen.)

The amino group is first protected with a typical protecting group such as t-butoxycarbonyl (hereinafter referred to as "BOC"), and then the carboxyl group of the amino acid thus protected in the presence of carbodiimide of formula (XIII) Reaction with an amine salt affords a compound of formula (XIV).

_{^{YO-NH + 3 Cl - (}} XIII)

Wherein Y is alkyl or benzyl.

Wherein Y is as defined above and at least one of R ₃ and R ₄ is hydrogen.

The hydroxyl groups of the general formula (XIV) compounds are converted to free groups (V) by reaction with typical reagents such as methanesulfonyl chloride (hereinafter referred to as methanesulfonyl as "Ms"). Other free groups (V) that can be used in this reaction include benzenesulfonyl, toluene sulfonyl, chloro, bromo and iodo.

The fully protected compound of formula (XV) is cyclized by treatment with a base such as, for example, potassium carbonate.

Wherein Y is as defined above and at least one of R ₃ and R ₄ is hydrogen.

This reaction is suitably carried out under reflux conditions in an organic solvent such as acetone, and this reaction generates a molar moiety of the following general formula (XVI).

Wherein Y is as defined above and at least one of R ₃ and R ₄ is hydrogen.

Alternatively, the cyclization of the general formula (XIV) compound may be carried out without first converting the hydroxyl group to the free group. The general formula (XIV) compound is treated with triphenylphosphine and diethylazodicarboxylate to prepare a general formula (XVI) compound wherein at least one of R ₃ and R ₄ is hydrogen.

Removal of the protecting group at the 1-position of the azetidinone of the general formula (XVI) can be carried out by a sodium reduction reaction when Y is alkyl, whereby an intermediate compound of the general formula (XVII) is obtained.

Wherein at least one of R ₃ and R ₄ is hydrogen.

If Y is benzyl, the catalyst (eg, active base palladium) hydrogenation yields the corresponding N-hydroxy compound initially, followed by treatment with titanium tricholide to ensure that at least one of R ₃ and R ₄ is hydrogen. Obtain an intermediate compound of formula (XVII).

Ring closure synthesis reactions of the type described above reverse the carrier chemical arrangement of the R ₃ and R ₄ substituents.

As described above, the azetidinone may be sulfonated to prepare a compound of the following general formula (XVIII).

Wherein at least one of R ₃ and R ₄ is hydrogen.

In another method for preparing compounds of formula (I) wherein R ₂ is hydrogen and at least one of R ₃ and R ₄ is hydrogen, the amino acid amide of formula (XIX) is used as starting material.

Wherein at least one of R ₃ and R ₄ is hydrogen.

Compounds of formula (XX) are prepared by protecting an amino group with a representative protecting group such as benzyloxycarbonyl (hereinafter referred to as "Z") or Boc and converting the hydroxyl group to a free group (V) such as Ms. .

Wherein at least one of R ₃ and R ₄ is hydrogen and A is a protecting group.

Formula (XXI) compounds are prepared by sulfonating Formula (XXI) compounds.

Wherein at least one of R ₃ and R ₄ is hydrogen.

Cyclization of the general formula (XXI) compound is carried out by treatment with a base (eg potassium carbonate). This reaction is suitably carried out under reflux conditions in a mixture of water and an organic solvent (eg, halogenated hydrocarbons such as 1, 2-dichloroethane), and this reaction gives a compound of the general formula (XXII).

Wherein at least one of R ₃ and R ₄ is hydrogen.

The sulfonated azetidinone of general formula (XXII), in which A is a protecting group, and the aforementioned counterpart compound having an R ₂ -alkoxy group are deprotected by catalytic hydrogenation to prepare a compound of general formula (XXIII).

Wherein at least one of R ₃ and R ₄ is hydrogen and R ₂ is hydrogen or an alkoxy group which can be converted to the corresponding zwitterion of formula (XXIV) by treatment with an acid such as formic acid.

Wherein at least one of R ₃ and R ₄ is hydrogen.

The sulfonated azetidinone of formula (XXII), in which A is a Boc protecting group, was removed under acidic conditions (e.g., using formic acid) to obtain the corresponding zwitterion of formula (XIV).

Good sources of β-lactam starting materials are 6-aminophenicylic acid and 7-aminocephalosporanic acid, each of which may optionally have 6-alkoxy or 7-alkoxy substituents. The general formula of these compounds is as follows.

(Wherein R ₂ is hydrogen or alkoxy and R ₁ is hydrogen or acyl)

3-amino-2-azetidinone can be prepared by the method described in the literature, for example by Chem. Soc. Special Publication No. 28, 288 (1977), The Chemistry of Penicillins (257) and Synthesis 494 (1977), published by Princeton University.

The 6-amino-phenicylic acid or 7-aminocephatosporanic acid is initially recaptured by reduction with Raney-nickel. This reaction can be carried out in water under reflux conditions, whereby a compound of the general formula (XXXVI) is produced.

₃ of Formula (I), wherein R ₁ is hydrogen or acyl, R ₂ is hydrogen or lower alkoxy, and R ₃ and R ₄ are hydrogen -Amino-3-alkoxy-2-azetidinone was obtained. Treatment of the general formula (XXVI) compound with cupric acetate and lead tetraacetate in an organic solvent (eg acetonitrile) replaces the carboxyl group with an acetate group. The resulting compound can be hydrolyzed using potassium carbonate in the presence of sodium borohydride.

The reaction intermediate is reacted with a complex of dimethylformamide and sulfur trioxide to introduce a sulfo group at the 1-position of the 3-amino-3-alkoxy-2-azetidinone compound.

The 3-azido-2-azetidinone starting material is first reacted with an olefin of the general formula (XXVII) with a halosulfonyl isocyanate of the general formula (XXVIII) (chlorosulfonyl isocyanate is suitable). Prepared by obtaining azetidinone.

Reduction hydrolysis of azetidinone of general formula (XXIX) yielded N-unsubstituted β-lactam of general formula (XXX).

A more detailed description of the reaction can be found in, for example, Chem. Soc. Rev., 5, 181 (1976) and J. Org. See Chem., 35, 2043 (1970).

3- of the azetidinone (or its sulfonated counterpart) is reacted with an arylsulfonyl azide (e.g., toluenesulfonyl azide) by reacting an azetidinone (or its sulfonated counterpart) of formula (XXX) By introducing an azido group at the position, the starting material azetidinone of the general formula (XXXI) can be obtained.

This reaction initially protects the azetidinone nitrogen with a silyl moiety (e.g., t-butyl dimethylsilyl or t-butyldiphenylsilyl) and then reacts the nucleus with a strong organic base (e.g., lithium diisopropylamine) at low temperature. It is most suitable to form an anion at the 3-position and then treat this anion with toluene sulfonyl azide. The resulting intermediate was quenched with trimethylsilyl chloride, and then the N-protecting group was subjected to acid hydrolysis or fluorine solubilization to obtain a compound of formula (XXXI).

Alternatively, compounds of formula (XXXI) can be prepared by reacting a primary amine having the general formula with a aldehyde of formula R ₃ CH = 0 to obtain the corresponding Schiff base.

or

-Azido acetic acid was reacted with the active form of [2 + 2] cyclic addition to obtain 3-azido-2-azetidinone of the following general formula.

또는

이다).(Where Q is

or

to be).

Oxidative removal of the Q-substituent yields a general formula (XXXI) compound.

Prepare by reducing 3-azido-2-azetidinone of Formula (XXXI) to give the corresponding 3-amino-2-azetidinone and then acylating this 3-amino-2-azetidinone. Can be.

As described when R ₂ is lower alkoxy, this product can be prepared from the corresponding compound in which R ₂ is hydrogen. Amide nitrogen of the nonalkoxylated compound is chlorinated to prepare an intermediate of formula (XXXII).

Reagents and processes required for N-chlorinated amide are known in the art. Representative reagents are tert-butylhypochlorite, sodium hypochlorite and chlorine. This reaction can be carried out in an organic solvent (eg lower alkanol such as methanol) or in a biphasic solvent system (eg water / methylene chloride) in the presence of a base such as sodium borate decahydrate. It is preferable to perform this reaction at low temperature.

The intermediate of formula (XXXI) can be reacted with an alkoxylating agent (eg, alkali metal alkoxide) to produce the product of formula (I) wherein R ₂ is alkoxy and enantiomers thereof. This reaction can be carried out in an organic solvent such as a polar organic solvent (eg dimethylformamide) at low temperature.

Alternatively, the compound of formula (I) wherein R ₂ is alkoxy is initially an intermediate of formula (VI) wherein R ₁ NH is cartbamate (eg, R ₁ is benzyloxy carbonyl) and R ₂ is hydrogen Can be prepared by alkoxylation, followed by introduction of a sulfo group at the 1-position of the resulting compound.

The intermediate compound of the general formula (XXXIII) was prepared by chlorinating the compound of the general formula (VI) using the above-described method [method of chlorinating the non-alkoxylated compound of the general formula (I) to prepare a compound of the general formula (XXXII)]. Got it.

The above-mentioned alkoxylation method [method of converting the compound of formula (XXXII) to the product of formula (I)] is used, and then a reducing agent such as trimethylphosphite is added to form a compound of formula (XXXIII). ) And its enantiomers.

By this process, the product of formula (I) wherein R ₂ is alkoxy is obtained in the form of a racemic mixture. If necessary, the enantiomer having the R stereo configuration can be fractionally crystallized using a suitable salt with an optionally active organic amine or racemic using conventional methods such as ion pair chromatography using optionally active cations. It can be isolated from the mixture.

Biological Preparation of Antibiotic EM5117

In the American Type Culture Collection, ATCC No. (R) -3- (acetylamino) -3-methoxy-2-oxo-1-azetidinesulfonic acid by culturing the microorganism Chromobacterium violaceum SC 11, 378 strain deposited at 31532 Salts (in Formula I, wherein R ₁ is acetyl and R ₂ is a methoxy group, hereinafter referred to as EM 5117) can also be prepared.

microbe

The microorganism used to prepare EM5117 is the Chromobacterium bioraceum SC 11, 378 strain. The subculture of microorganisms can be obtained from strains permanently deposited in The American Type Culture Collection, Rockville, Maryland. The accession number of this strain is heading 31.322. In addition to the specific microorganisms described above, EM5117 can also be produced by culturing a mutant of the microorganism (for example, a mutant by X-rays, ultraviolet radiation or nitrogen mustards).

Chromobacterium bioraceum SC 11, 378 A.T.C.C. Heading 31.32 can be isolated from the moist sample soil containing this microorganism by first stamping the sample soil in a medium containing the following material.

Soil Extract 400ml Glucose 10.0g

Distilled Water 600ml Agar 17.5g

Yeast Extract 5.0g

The medium was adjusted to pH 6.0 and sterilized for 20 minutes in an autoclave at 121 ° C. Colonies of Chromobacterium violaseum SC 11, 378 are incubated at 25 ° C. for 24 to 72 hours and then isolated from plate soil. The isolated cells were then incubated in a medium containing the following material.

Yeast Extract 1g Glucose 10g

Beef Extract 1g Agar 15g

2 g of NZ Amine A to 1 l of distilled water

The medium was adjusted to pH 7.3 and autoclaved at 121 ° C. for 30 minutes.

Chromobacterium bioolaseum SC11, 378 is a Gram-negative rod that frequently exhibits lateral, bipolar straining and lipid inclusions. These microorganisms move into unipolar flagella and sometimes small sized side flagella.

In nutrient agar, Chromobacterium bioraceum SC 11, 378 produces purple colonies. The pigment becomes thicker in a medium containing enough tryptophan and yeast extract. In the nutrient solution, a purple ring is formed on the wall of the test tube, but no fusion film is formed. The purple pigment is soluble in ethanol but insoluble in water and chloroform.

Chromobacterium violaseum SC 11, 378 is a mesophilic bacterium, so it develops at 15 ° ~ 37 ℃ and does not grow at 4 ℃ or 37 ℃. This microorganism is an oxidase positive that hydrolyzes casein well. Glucose, fructose and trehalose are fermented in the presence of Chromobacterium bioraceum SC 11,378 (see Hugh & Leifson method, 1953). L-arabinose is not oxidized or fermented by this microorganism. This microorganism produces hydrogen cyanide and is negative for aesculin hydrolysis.

The main characteristic is the basis for distinguishing the Chromobacterium violaseum from Chromobacterium lividum, which is the only other genus in the Chromobacterium recognized by Bergey's Manual of Determinative Bacteriology 8th edition. Species.

EM5117 can be prepared by culturing other strains of Chromobacterium bioraceum.

Antibiotic

In order to obtain antibiotic EM5117, chromobacterium bioraceum SC 11, 378 A.T.C.C. No. 31,322 was incubated at about 25 DEG C while blowing air into a nutrient medium containing carbon and nitrogen sources capable of assimilation. Fermentation is usually carried out for about 18 to 24 hours (approximately 20 hours are suitable) until substantial antimicrobial activity is imparted to the medium.

EM5117 was purified from the fermentation broth by the following procedure. After fermentation, the cells were centrifuged or filtered to remove the mycelium from the fermentation broth. After removing the mycelium from the fermentation broth, fermentation of EM5117

It can extract from liquid. The fermentation broth is preferably extracted by ion pair extraction with cetyldimethyl benzylammonium chloride in methylene chloride at pH 5 and back extracted in aqueous sodium iodide (adjusted to pH 5 with acetic acid). After concentrating the sodium iodide extract in vacuo, the aqueous solution of the residue can be washed with butanol. The resulting aqueous solution was concentrated to dryness and the residue obtained was dissolved in methanol as much as possible. Insoluble matters were separated by centrifugation and the insolubles were washed with methanol and then discarded.

The methanol concentrate was dissolved in methanol / water (1: 1) and the antibiotics were purified using a column containing a chromatography column, such as Sephadex G-10, in the same solvent mixture. After eluting with the same solvent mixture, the effective fractions were combined and concentrated to dryness. The residue was mixed with methanol and the insoluble material was filtered off.

For further purification, the methanol soluble material was purified on a DEAE cellulose column. The column can be eluted with a gradient in 0.1 M sodium phosphate buffer at pH 5. The effective fractions were combined, concentrated and the methanol insolubles were removed.

To further purify EM5117, this material is dissolved in water and then run on a column of Sephadex LH-20, which is then eluted with water. The effective fractions were combined and concentrated. Further purification of EM5117 is performed by dissolving this material in water and then eluting it with water by developing this solution on a column of Diionion HP-20AG. The effective fractions were combined and concentrated. This concentrate was dissolved in water, passed through a potassium type Dowex 50W-X2 column and washed with water. The effluent was concentrated to give a relatively pure potassium salt of EM5117 as crystalline material.

By the above-described isolation and purification process, the potassium salt of EM5117 was produced. Other salts can be prepared according to the type of ion exchange resin used in the final purification step.

Biological Preparation of Antibiotic EM5210

Various acetic acid bacteria were cultivated and R _1-3 -D [N- (D-γ-glutamyl) of general formula (I) wherein R ₁ is D-γ-glutamyl-D-alanyl and R ₂ is methoxy. ) -D-alanyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid (hereinafter referred to as EM5210) salts can be prepared.

microbe

The microorganism used to prepare EM5210 is the Gluconobacter SC11,435 strain. The subculture of this microorganism can be obtained from a strain permanently deposited in the American Type Culture Collection, which is introduced in Rockville, Maryland. The accession number of this strain is A.T.C.C. No. 3158. In addition to the specific microorganisms described above, EM5210 can also be produced by culturing a mutant of the microorganism (for example, a mutant by X-ray, ultraviolet radiation or nitrogen mustard).

Gluconobacter SC11,435, A.T.C.C. No. 31581 can be isolated from the above ground moss by first incubating the above ground moss containing microorganisms in a 10% aqueous solution of pectin (pH2.5) at 25 ° C. for 7 days. This microorganism can be isolated by plating on a medium containing the following materials.

Extracts of the Spartina Parthen grass were prepared by adding 3 l of tap water to 500 g of chopped and dried grass, and boiled for 30 minutes. The medium was adjusted to pH 6.0 and sterilized for 20 minutes in an autoclave at 121 ° C. After incubation at 25 ° C. for about 18 hours, the cells of Gluconobacter SC11,435 were isolated from the platen pecton nutrient solution. The isolated colonies were then incubated in a medium containing the following materials.

The medium was adjusted to pH7.3 and autoclaved at 121 ° C. for 30 minutes.

Gluconobacter SC11,435 is a polymorphic Gram-negative rod that runs with one to three polar flagella. This bacterium is absolutely aerobic, catalase positive and oxidative. It is distinguished from Pseudomonas in that it is cytochrome oxidase negative and extremely acid resistant. These properties indicate that the microorganisms are more associated with acetic acid bacteria than true Pseudomonas.

Gluconobacter SC11,435 in BBL tryticase soy agar grows as a mixture of coarse and soft colonies. Coarse colonies are associated with a pale yellow soluble pigment, while soft colonies are spotted and colorless. The separation in two forms is due to the influence of medium, temperature and storage conditions. The activity of EM5210 tends to decrease in many cultures with coarse colonies.

When Glunobacter SC11,435 is incubated in BBL Watt agar (pH4.8), the sticky colonies of the spots grow lushly. Similar growths were obtained with malt-yeast extract agar (1% each) adjusted to pH4.5.

Sufficient acid was generated from glucose on a medium containing 1% yeast extract, 10% glucose, 3% calcium carbonate and 2.5% agar to form a portion of the calcium carbonate disappearance around the bacterial growth. This is the hallmark of Acetobacter and Gluconobacter.

Yeast extract-glycerol and yeast extract-calcium lactate in the presence of Gluconobacter SC11,435 shows a brown soluble pigment on agar plates, and glucose, fructose, galactose, mannose, xylose, mannitol and arabinose produce acid but Mixing lactose, sucrose or maltose in Hugh and Leifson medium neither grows nor produces acids.

Microorganisms capable of biologically producing EM5210 are not limited to Gluconobacter SC11,435 but can be used as the acetic acid bacteria. The following cultures can be used to prepare EM5210, respectively.

Acetobacter Pasteurians Subspecies Pasteurianium A.T.C.C. 6033

Acetobacter aceti subspecies Aceti A.T.C.C. 15973

Gluconobacter Oxidans Subspecies Oxidans A.T.C.C 19357

Gluconobacter oxydans subspecies suboxydans A.T.C.C. 23773

Gluconobacter Oxidans Subspecies Oxidans A.T.C.C. 15178

Acetobacter Aceti subspecies liquefaciens A.T.C.C. 23751

Acetobacter peroxidans A.T.C.C. 12874

Gluconobacter oxydans subspecies suboxydans A.T.C.C. 19441

Acetobacter spp. A.T.C.C. 21780

Gluconobacter Oxidans Subspecies Industrialis A.T.C.C. 11894

Antibiotic

Gluconobacter SC11,435 A.T.C.C. Antibiotic EN5210 may be prepared by incubating at No. 31581 or one of the above-mentioned microorganisms at about 25 ° C. while blowing air in a nutrient medium containing an anabolic carbon and a nitrogen source. Fermentation is usually carried out for about 16 hours 24 hours (approximately 20 hours are suitable) until substantial antimicrobial activity is imparted to the medium.

By the following process, EM5210 was isolated from the fermentation medium and purified. After the fermentation was finished, the fermentation broth was centrifuged to remove bacteria, and the supernatant of the fermentation broth was adsorbed on an anion exchange resin (eg, Dowex 1X-2) at pH 3.7 to separate antibiotics. This antibiotic was eluted from the resin with sodium chloride at pH 4 and the eluate was concentrated and passed through an activated carbon column. EM5210 was then eluted from methanol: water (1: 1) from activated carbon. The effective fractions were combined, dried and placed in an anion exchange column (eg Dowex 1-X2) and then eluted with sodium chloride buffered to pH4. The effective fractions were collected and desalted in macroreticular styrene-divinylbenzene copolymer resin (Dianion HP 20AG). The effective fraction eluted with water was concentrated and freeze dried to obtain the sodium salt of EM5210.

The sodium salt of EM5210 is produced by the above purification process. Other salts can be obtained by changing the salt used to elute the EM5210 in ion exchange chromatography. For example, potassium chloride is used to obtain potassium salt.

The following examples are intended to specifically illustrate the present invention.

Example 1

(S) -N- (2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide potassium salt

Method I

A) 1-[(1R) -carboxy-2-methyl (propyl)]-2-oxo (3S)-[phenyl [acetyl (amino)]] azetidine

Raney nickel was decanted with water for several hours until the pH of water (5-6 times the volume of Raney nickel) reached 7.6.

To a solution of 9.0 g of penicillin G (Na ⁺ ) in 500 ml of water, 54 g (90 ml) of Raney nickel was added. The flask equipped with the reflux condenser was immersed in a bath at 155 ° C. At the start of reflux, this reflux was continued for 15 minutes. The flask was quenched in an ice-water bath and filtered through Celite to remove Raney nickel. The pH was adjusted to 3 with dilute HCl, and the aqueous solution was concentrated to about 150 ml and then cooled. The oily layer was crystallized after scratching. After washing with water and drying in vacuo at 50 ° C. for 3 hours, 3.83 g of the target compound were obtained.

B) 1- [acetyloxy-2-methyl (propyl)]-2-oxo- (3S)-[phenyl- [acetyl (amino)]] azetidine

Nitrogen was injected while bubbling the solution for 15 minutes while stirring the solution of 608 mg (2 mmol) of the compound in 20 ml of anhydrous acetonitrile. All acids were dissolved for a few minutes in a water bath at 40 ° -45 ° C.

The water bath was removed and powdered copper diacetic acid dihydrate (182 mg, 1 mmol) was added, followed by stirring for 1 minute, followed by 886 mg (2 mmol) of lead tetraacetate. The mixture was stirred at room temperature for 20 minutes. The acetonitrile solution was poured over and the precipitated solids were washed with ethyl acetate. The combined acetonitrile-ethyl acetate solution was evaporated until a residue remained, and the residue was dissolved in ethyl acetate-water. The ethyl acetate layer was washed three times with water, once with aqueous sodium bicarbonate solution (pH7), and once with water. The ethyl acetate layer was dried over sodium sulfate, evaporated until a residue (515 mg) remained, and the residue was used without further purification in the next step of reaction.

C) 2-oxo- (3S)-[phenyl [acetyl (amino)]] azetidine

To a solution of 911 mg (2.86 mmol) of the compound in 21 ml of methanol was added 3.5 ml of water, followed by 383 mg of potassium carbonate (2.86 mmol).

The mixture was stirred for 1 minute in the presence of nitrogen and then 160 mg (4.30m mol) of sodium borohydride were added. The reaction was stirred at rt for 20 min. Methanol was removed in vacuo and the residue was dissolved in ethyl acetate and a small amount of water. The pH of this solution was adjusted to 2.5 degrees. The ethyl acetate layer was washed with an aqueous sodium bicarbonate solution at pH 7 and then with a small amount of water, finally dried over sodium sulfate and evaporated to give 493 mg of crude product. A small amount of ethyl acetate was added to give 250 mg of the desired crystalline product (yield 43%). By crystallization or chromatography, an extra amount of product can be obtained.

D) (S) -N- (2-oxo-1-sulfo-3-azetidinyl) -2-phenyl-acetamide, potassium salt

Pyridine SO ₂ complex (215 mg, 1.35 m mol) was added to a solution prepared by stirring 251 mg (1.23 m mol) of the above product in anhydrous dimethylformamide and 2 ml of anhydrous methylene chloride at room temperature in the presence of nitrogen. The mixture was stirred for 3 hours, then the solvent was removed in vacuo and the residue was dissolved in methylene chloride-water. The pH was adjusted to 6.5 with 2N potassium hydroxide and the aqueous layer was washed three times with methylene chloride and then evaporated until a residue was obtained. 20 ml of methanol was added to the residue, followed by stirring and filtration to remove potassium sulfate. 10-15 ml of methanol was added to the residue obtained by evaporation of the filtrate and stirred. The obtained solid was 49 mg of the target compounds of melting | fusing point 189 degreeC (decomposition).

This compound exhibited the same spectral properties as the product prepared by Method II.

Method II

A solution of 660 mg of (S) -2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid potassium salt in 13 ml of water was added together with 200 mg of 10% palladium based on activated carbon under hydrogen atmosphere. Stir for 2 hours. The catalyst was filtered off and the filtrate was diluted with the same volume of acetone and then cooled in an ice bath. After 30 minutes, phenylacetyl chloride (eight 40 μl each) and 10% potassium bicarbonate solution were added. (keep pH between 5.2 and 5.8). After 40 minutes, the solution was concentrated in vacuo to remove acetone and the solution was developed in a 200 ml HP-20 column. After eluting with water and then with water-acetone (9: 1), the Lydon-positive fraction was subjected to TLC test, pooled and evaporated to give 160 mg of crude product. Crystallization was carried out using methanol ether to obtain 101 mg of the target compound having a melting point of 210 ° C. (decomposition).

Method III

In a solution of 121 mg of (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) dissolved in anhydrous methylene chloride (3 ml), 40 mg of phenylacetic acid and dicyclohexylcarbodie 61 mg of mead was added. The mixture was stirred at rt for 48 h and then filtered to remove dicyclohexylurea. The solvent was removed in vacuo, the residual compound was dissolved in acetone, filtered, and 5 ml of acetone saturated with potassium iodide was added to precipitate the target compound. The supernatant was decanted off and the residue was washed three times with acetone and then dried to give 48 mg of product having the same spectral properties as the products of Methods I and II.

Method IV

A solution obtained by dissolving 2.83 g of (S) -2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and pyridine salt (see Example 2) in 36 ml of water was prepared from activated carbon. It was stirred under a hydrogen atmosphere with 707.5 mg% palladium (175 ml of hydrogen dissolved). After 2 hours, the slurry was filtered, the filtrate was cooled to 0 ° C. and then diluted with 46 ml of acetone (the initial pH was 4.25, but the pH was adjusted to 6.7 by addition of a cooled 10% potassium bicarbonate solution). A 2.4 ml solution of phenylacetyl chloride dissolved in 10 ml of acetone was added dropwise over 15 minutes. The cooled 10% potassium bicarbonate solution was added simultaneously to maintain the pH between 5.2 and 5.8. After 45 minutes, the slurry was diluted with 93 ml of 0.5 M potassium phosphate (pH = 4.2) and concentrated to remove acetone. The slurry was filtered and washed with water. The filtrate and washings were combined and eluted successively with 1 liter of 0.5 M potassium phosphate (pH = 4.2), 1 liter of water, and 2.5 liters of water-acetone (9: 1) using HP-20 column for 450 ml. 15 fractions were 200 ml and 16-21 fractions were 100 ml) to obtain 1.285 g of the target compound. The spectrum of the compound obtained by this method showed the same spectral data as the product obtained by the above method.

Example 2

(S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, pyridine (1: 1) salt

Method I

A) 1-[(1R) -carboxy-2-methyl (propyl)]-2-oxo- (3S)-[[(phenylmethoxy) carbonyl] amino] azetidine

5.18 g of sodium bicarbonate was added to 140 ml of water, and the slurry was added to 12.98 g (0.06 mol) of 6-aminophenic silane acid, which was stirred for about 10 minutes. Raney nickel suspension (washed with water to pH8.0, 260 ml of slurry is 130 g) was added in one portion. After 15 minutes the slurry was cooled, filtered and the filtrate was treated with a solution of 11.94 g (0.07 mol) of benzylchloro formate dissolved in 5.18 g of sodium bicarbonate and 12 ml of acetone. After 30 minutes, the solution was acidified to pH2.5 and extracted with methylene chloride. The organic layer was dried, evaporated and titrated with ether-hexane to give 6.83 g of the target compound.

B) 1-[(acetyloxy) -2-methyl (propyl)]-2-oxo- (3S)-[[(phenylmethoxy) carbonyl] amino] azetidine

The solution of 6.83 g (0.0213 mol) of acid dissolved in 213 ml of acetonitrile was treated with 1.95 g (0.0107 mol) of cupric acetate monohydrate and 9.5 g (0.0213 mol) of lead tetraacetate. The slurry thus obtained was immersed in an oil bath at 65 ° C., and the slurry was stirred while injecting nitrogen into the slurry while all starting materials reacted and consumed. The slurry was filtered and the solid obtained was washed with ethyl acetate. After the filtrate and the tax solution were combined and evaporated in vacuo, the residue was dissolved in 100 ml each of ethyl acetate and water, and the pH of this solution was adjusted to 7. The ethyl acetate layer was separated and dried evaporated to give 6.235 g of the target compound.

C) (S)-(2-oxo-3-azetidinyl) carbamic acid, phenylmethyl ester

A solution of 3.12 g (0.0093 mol) of the acetate dissolved in 70 ml of methanol and 7 ml of water was cooled to −15 ° C., followed by 1.33 g of potassium carbonate and 349 mg of sodium borohydride. The reaction mixture was stirred at -15 ° to 0 ° C. After completion of the reaction (about 2 hours), 2N HCl was added to the mixture to neutralize to pH7, and the mixture was then concentrated in vacuo. The concentrate was adjusted to pH5.8, saturated with salt, and extracted three times with ethyl acetate. After drying the organic layer, the residue produced by evaporation in vacuo was combined with the material obtained in a similar experiment and then titrated with ether to give 3.30 g of the target compound.

D) (S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, pyridine salt

Method I

The azetidi dissolved in 2 ml of anhydrous methylene chloride and 2 ml of anhydrous dimethylformamide was stirred with a 440 mg (0.002 mol) solution of 350 mg (0.00222 mol) of pyridine-3 sulfur oxide complex in the presence of nitrogen for 2 hours. Most of the solvent was removed in vacuo and the residue was titrated with ethyl acetate to give 758 mg of solid target compound.

Method II

Chlorosulfonyltrimethylsilyl ester (18.87 g) was added dropwise to 7.9 g of anhydrous pyridine with stirring at -20 ° C under a nitrogen atmosphere. After the addition was complete, the mixture was further stirred at room temperature for 30 minutes, and then trimethylchlorosilane was removed in vacuo. A solution of 20 g of the azetidinone (methods I and C) dissolved in 120 ml of dimethylformamide and 120 ml of methylene chloride was added and stirred at ambient temperature for 3.5 hours. The solvent was evaporated in vacuo to crystallize by adding ethyl acetate to the resulting oily residue, to give 31 g of the target compound. NMR data of this compound were identical to those of Method I.

Example 3

(S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

Method I

(S) -2-Corn- (3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, pyridan salt (see Example 2) 135 mg 0.5M potassium phosphate monobasic (2N potassium hydroxide) And pH eluted with 10 ml of buffer, 200 ml of water and 100 ml of acetone-water (1: 1) using a 25 ml HP-20AG column. This fraction was evaporated to give 80 mg of the desired compound having the same spectral data as the product of the following procedure.

Method II

600 mg of (S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid pyridine salt (see Example 2) was dissolved in 2 ml of water and monobasic at pH 5.5 Mix with 15 ml of potassium phosphate buffer. Once the solid had formed, the slurry was cooled to 0 ° C., filtered and washed successively with cold buffer, cold 50% ethanol, ethanol and ether to give 370 mg of the target compound (as a result of analysis, containing a large amount of potassium ions). Got it. A 280 mg solution of salt dissolved in 10 ml of water was eluted with 200 ml of water followed by water-acetone (9: 1) using an HP-20 column for 100 ml. Fractions (50 ml) were combined and this fraction 7 was evaporated to give a solid. After titration with acetone, filtration, and drying in vacuo, 164 mg of the target compound having a melting point of 193 ° C to 196 ° C was obtained.

Method III

20.0 g of (S)-(2-oxo-3-azetidinyl) carbamic acid and phenylmethyl ester (see Example 2C) are suspended in 200 ml of acetonitrile, to which monotrimethylsilyltrifluoroacetamide (25.3 g 21.6 ml) was added and the mixture was heated to 50 ° C. with stirring for 1 h. After cooling to 0 ° C. in an ice bath, 17.2 g of trimethylsilylchloro sulfonate was added dropwise and the solution was stirred at ambient temperature for 6 hours. To this solution was added 24.2 g of potassium hexanoate in 100 ml of butanol and further stirred for 1 hour. The slurry was poured into 1 l of anhydrous diethyl ether, and the precipitate was filtered off and dried in vacuo. The compound was dissolved in 500 ml of water, the pH was adjusted to 5.0 with potassium carbonate, and the insoluble material was filtered off. The mother liquor was freeze-dried to obtain 19.4 g of crude product. The compound contained a small amount of potassium chloride, but this potassium chloride was removed by chromatography. The obtained compound showed the same spectral data as the product of Method II.

Example 4

(S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt (1: 1)

Method I

34.3 g of (S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and pyridine salt (1: 1) (see Example 2) were dissolved in 800 ml of water. . This solution was purified with activated charcoal, to which 30.7 g of hydrogen tetrabutylammonium sulfate in 80 ml of water was added, and then the pH was adjusted to 5.5 with 1N potassium hydroxide. The solvent was removed in vacuo until the volume reached about 200 ml. The precipitated tetrabutylammonium salt was filtered off and dried in vacuo. The compound was recrystallized from water or dissolved in methylene chloride, filtered and precipitated by addition of tet. Yield 34.3 g, melting point 108-110 ° C.

Method II

20.2 g of (S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt (1: 1) (see Example 3) were dissolved in 500 ml of water After filtration, 20.3 g of tetrabutyl ammonium hydrogen sulfate in 100 ml of water were added. 1N potassium hydroxide was added thereto to adjust the pH to 5.5. Evaporate in vacuo until the volume is about 100 ml, and precipitated tetrabutylammonium salt is filtered off. This compound was dissolved in 30 ml of methylene chloride, filtered, and etet was added and precipitated to obtain 21 g of the target compound having a melting point of 109 to 111 ° C.

Example 5

(3S) -α-[[(2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] benzeneacetic acid, phenylmethyl ester potassium salt (1: 1)

A) (S) -3-amino-2-azetidinone

3 g of (S)-(2-oxo-3-azetidinyl) carbamic acid, phenylmethyl ester (see Example 2C) were hydrogenated in 100 ml of methanol in the presence of 1 g of palladium catalyst based on activated carbon. When the theoretical amount of hydrogen was absorbed, the catalyst was filtered off and the filtrate was evaporated and dried. This was left to obtain 1.1 g of the target compound as crystals.

B) (3S) -α-[[(2-oxo-3-azetidinyl) amino] carbonyl] benzeneacetic acid, phenylmethyl ester

The azetidinone (3.0 g) was dissolved in 100 ml of dimethylformamide, the solution was cooled to 0 ° C., and then 4.5 g of N-methyl morpholine was added (dropped), followed by α- (chloro in 50 ml of acetonitrile. Carbonyl) benzeneacetic acid and 10.8 g of phenylmethyl esters were added dropwise while stirring. The mixture was stirred at 5 ° C. for about 16 hours, the solvent was distilled in vacuo and then 100 ml of water was added to this residue. The obtained suspension aqueous solution was extracted twice with 100 ml of methylene chloride. The organic layers were combined, washed successively with sodium bicarbonate, 2N phosphoric acid and water, dried over sodium sulfate, filtered and evaporated to dryness. This residue was crystallized with ethyl acetate and petroleum ether to obtain 8.7 g of the target compound having a melting point of 164 ° C to 166 ° C.

C) (3S) -α-[[(2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] benzeneacetic acid, phenylmethyl ester potassium salt (1: 1)

The compound (6.9 g) was suspended in 150 ml of acetonitrile, to which monotrimethylsilyltrifluoroacetamide (5.7 g) was added, and then the solution was heated with stirring at 50 ° C. for 30 minutes. The solution was cooled to 0 ° C. and 3.9 g of trimethylsilylchlorosulfonate was added dropwise. After the addition was completed, the mixture was heated to 50 ° C. for 5 hours. After cooling to 20 ° C., 7.6 g of potassium hexanoate ethyl in 10 ml of butanol were added and stirred for 30 more minutes. When 300 ml of ether was added, the desired compound precipitated, which was filtered off. 100 ml of anhydrous acetonitrile was added to the crude product, stirred for 30 minutes, and filtered to obtain 4.5 g of the target compound having a melting point of 118 ° to 120 ° C. The crude product was further purified by HP20 chromatography, and then lyophilized to obtain a pure material having a melting point of 188 ° to 190 ° C.

Example 6

(S) -3-[[(2-amino-4-thiazolyl) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

A) (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

2 g of (S) -2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 4) were dissolved in 100 ml of dimethylformaldehyde as a catalyst. 1 g of activated carbon based palladium (10%) was hydrogenated for about 30 minutes. The catalyst was filtered off and dimethylformamide was removed to give an oily target compound.

NMR (CDCl ₃ )

3.82 (1H, t, J = 5.5)

4.05 (d, 1H, dd, J = 5.5,2.5cps)

B) (S) -3-[[(2-amino-4-thiazolyl) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

0.7 g solution of dicyclohexylcarbodiimide dissolved in 2 ml of the compound, 0.5 g of aminothiazole acetic acid and 0.4 g of hydroxybenzothiazole in 100 ml of anhydrous dimethylformamide, and then stirred at 0 ° C., in 10 ml of dimethylformamide. Was added drop wise. After addition, stirring was continued at 20 ° C. for 12 hours. The insoluble urea was filtered off, and the oily residue obtained by evaporation of the solvent in vacuo was treated with potassium sulfate perfluorobutane solution dissolved in 20 ml of acetone for 15 minutes at room temperature. After addition of 200 ml of dimethyl ether, the desired compound precipitated, which was filtered and evaporated and then purified through a 300 ml HP-20 chromatography column (using water as eluent). 850 mg of the target compound having a melting point of 300 ° C. or higher was obtained.

Example 7

[3S (±)]-3-[[(formyloxy) phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid potassium salt

1.5 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutyl ammonium salt (see Example 6A) in 100 ml of dimethylformamide and 2 ml of propylene oxide were cooled to 0 ° C. To this, a solution of 0-propylmandelic acid chloride dissolved in 10 ml of acetonitrile was added dropwise while stirring. After maintaining this temperature for 1 hour, the solvent was distilled off in vacuo. The oily residue was treated with a solution in 2 g of potassium sulfate perfluorobutane dissolved in 15 ml of acetone. After 20 ml of ether was added thereto, the target compound crystallized and this was filtered to obtain 1.5 g of the product. This product was purified by HP-20 chromatography.

Melting Point 180 ° ~ 185 ° C (Decomposition)

Example 8

[3S (+)]-3-[[(formyloxy) phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

The procedure of Example 7 was followed except that D-0-formyl mandelic acid chloride was used instead of only 0-formylmandelic acid chloride, and lyophilized to obtain the target compound having a melting point of 120 to 125 ° C.

Example 9

[3S (1)]-3-[[(formyloxy) phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

The procedure of Example 7 was followed except that LO-formylmandelic acid chloride was used instead of O-formylmandelic acid chloride to obtain a target compound having a melting point of 203 ° to 205 ° C containing 1 mole of water. . After carefully drying, a product having a melting point of 228 ° to 230 ° C. was obtained.

Example 10

(S) -2-oxo-3-[(1-oxooctyl) amino] -1-azetidinesulfonic acid, potassium salt

1.5 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) in 100 ml of dimethylformamide and 2 ml of propylene oxide were cooled to 0 ° C. At this temperature, a solution of 0.8 g of caprylic acid chloride dissolved in 20 ml of anhydrous acetone was added dropwise and stirred for 30 minutes. The solvent was evaporated in vacuo and the oily residue was treated with 2 g of potassium sulfate perfluorobutane in 15 ml of acetone. Acetone was distilled in vacuo, and the residue was dissolved in 5 ml of water, then eluted with 300 ml of HP-20 resin and water / acetone (9: 1) as eluent, melting point 173 ° -180 ° C. (after freeze drying). 0.9 g of the desired compound was obtained.

Example 11

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[[hydroxy (phenylmethoxy) phosphinyl] methoxy] imino] acetyl] amino] -2-oxo-1 Azetidinesulfonic acid, potassium salt

(S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) in 30 ml of dimethylformamide 0.8 g, (Z) -2-amino-α-[[[hydroxy 0.9 g of (phenylmethoxy) phosphinyl] methoxy] imino] -4-thiazole acetic acid, 0.3 g of hydroxybenzotriazole and 0.7 g of dicyclohexylcarbodiimide were stirred at room temperature for 24 hours. The precipitated urea was filtered off and the solvent was removed in vacuo. The remaining oil was treated with 1 equivalent of potassium sulfate perfluorobutane in 10 ml of acetone. The target compound was filtered and purified using HP-20 resin and water as eluent to obtain 500 mg of the target compound having a melting point of 210 ° to 215 ° C. (decomposition).

Example 12

[3S (Z) -3-[[(2-amino-4-thiazolyl) (ethoxyimino) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

(S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) 1.5 g, hydroxybenzothiazole 0.6 g, 1 g dicyclohexylcarbodiimide in 100 ml of dimethylformamide And 0.8 g of (Z) -2-amino-α- (ethoxyimino) -4-thiazole acetic acid were stirred at room temperature for 24 hours. The solvent was distilled off and the residue was dissolved in 30 ml of acetone. Urea was filtered off and the mother liquor was treated with a solution of 2 g of potassium sulfate pifluoro part dissolved in 20 ml of acetone. After 200 ml of ether was added thereto, the desired compound was precipitated, which was filtered off and dried. Purification by chromatography using HP-20 column and water as the eluent gave 1.1 g of the target compound having a melting point of 180 ° -185 ° C (decomposition).

Example 13

[3S (E)]-3-[[(2-amino-4-thiazolyl) (ethoxyimino) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

It is carried out except using (E) -2-amino-α- (ethoxyimino) -4-thiazole acetic acid instead of (Z) -2-amino-α- (ethoxyimino) -4-thiazole acetic acid. It carried out according to the method of Example 12 and obtained the target compound of melting | fusing point 160 degreeC-170 degreeC (after freezing drying).

Example 14

[3S (Z)-3-[[(2-amino-4-thiazolyl] (2,2,2-trifluoroethoxy) imino] acetyl] amino] -2-oxo-1-azetidine Sulfonic acid, potassium salt

(Z) -2-amino-α-[(2,2,2-trifluoroethoxy) imino] instead of (Z) -2-amino-α- (ethoxyimino) -4-thiazole acetic acid Except for using the 4-thiazole acetic acid, it carried out according to the method of Example 12, and obtained the target compound of melting | fusing point 160 degreeC-170 degreeC (after freezing drying).

Example 15

(S) -2-oxo-3-[(1-oxopropyl) amino] -1-azetidinesulfonic acid, potassium salt

Method I

1.5 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) in 100 ml of anhydrous dimethylformamide and 4 ml of propylene oxide were cooled to 0 ° C. under stirring. At this temperature, 0.5 g of propionic acid chloride in 10 ml of acetonitrile was added dropwise and the mixture was stirred for 2 hours. The solvent was distilled in vacuo and the oily residue was treated with 1 equivalent of potassium sulfate perfluorobutane in 5 ml of acetone. After adding the ether, the target compound was crystallized, and the resultant was filtered to obtain 0.8 g of the target compound having a melting point of 135 ° to 140 ° C. (after freeze drying).

Method II

In 100 ml of diglyme, 4 g of (S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidine sulfonic acid and tetrabutylammonium salt (see Example 4) was replaced with 1 g of palladium based on activated carbon. Added. After 2.5 hours, hydrogenation was terminated. The catalyst was filtered off, 2 ml of propylene oxide was added followed by cooling to 0 ° C. and 0.5 g of propionic acid chloride in 10 ml of anhydrous diglyme was added with stirring. After 30 minutes the solvent was removed in vacuo and the oily residue was treated with 1 equivalent of potassium sulfate perfluorobutane in 20 ml of acetone. After the addition of ether, the desired compound was crystallized, which was filtered and then recrystallized from water / acetone to give 0.9 g of a product having a melting point of 156 ° -160 ° C. (decomposition).

Example 16

[3S (±)]-3-[(hydroxyphenylacetyl) amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

1.5 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) in 100 ml of anhydrous dimethylformamide, 1.5 g of dicyclohexylcarbodiimide, hydroxybenzotriazole 0.5 g and 0.6 g mandelic acid were stirred for about 16 hours. The solvent was removed in vacuo and the residue was dissolved in 20 ml of acetone. The precipitated urea was filtered off and the mother liquor was treated with 1 equivalent of potassium sulfate fifluorobutane. After adding ether, the target compound precipitated, and the resultant was filtered to obtain 1.4 g of crude product. After recrystallization using water, a product having a melting point of 138 ° to 140 ° C. was obtained.

Example 17

(S) -3-[[[((cyanomethyl) thio] acetyl] amino-2-oxo-1-azetidinesulfonic acid, potassium salt

(S) -3-amino-2-oxo-1-azetidinesulfonic acid, 1.5 g of tetrabutylammonium salt (see Example 6A) and 0.72 g of [(cyanomethyl) thio] acetic acid were dissolved in 70 ml of acetonitrile, and To a solution of 1.04 g of dicyclohexylcarbodiimide dissolved in acetonitrile was added dropwise. The mixture was stirred at 0 ° C. for about 16 hours, the precipitated dicyclohexylurea was filtered off, the filtrate was evaporated and the oily residue was dissolved in acetone. To this, a saturated potassium iodide solution dissolved in acetone was added to precipitate the target compound to obtain 1.1 g of a product having a melting point of 150 ° to 155 ° C.

Example 18

(S) -2-oxo-3-[(1H-tetrazol-1-ylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

0.77 g of 1H-tetrazol-1-acetic acid in a 0.005 mole solution of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 6A) dissolved in 70 ml of dimethylformamide; And a solution of 1.13 g of dicyclohexylcarbodiimide dissolved in 5 ml of dimethylformamide were added. The mixture was stirred at rt for about 16 h and then the solvent was removed in vacuo. The remaining oily substance was dissolved in 20 ml of acetone and treated with a 0.006 mol solution of potassium sulfate perfluorobutane dissolved in acetone to obtain 1.5 g of the target compound having a melting point of 170 ° to 175 ° C (decomposition).

Example 19

(S) -2-oxo-3-[(2H-tetrazol-2-ylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

Except for using 2H-tetrazol-2-acetic acid instead of 1H-tetrazol-1-acetic acid, it carried out according to the method of Example 18, and obtained the target compound of melting | fusing point 175 degreeC-177 degreeC (decomposition).

Example 20

(S) -2-oxo-3-[(2-thienylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

A target compound having a melting point of 180 ° to 190 ° C. (decomposition) was obtained in accordance with the method of Example 18 except that 2-thiophenacetic acid was used instead of 1H-tetrazol-1-acetic acid.

Example 21

[3S (Z)]-3-[[(2-amino-4-thiazolyl) (methoxyimino) -acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

As described in Example 6A, (S) -3 prepared from 7.9 g of (S) -2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt -Amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt are cooled to 0 degreeC, and 3.53 g of (Z) -2-amino- (alpha)-(methoxyimino) -4-thiazole acetic acid are added here. Then, a solution of 3.27 g of dicyclohexyl-carbodiimide dissolved in 10 ml of dimethylformamide was added. The mixture was stirred at 5 ° C. for 16 h, filtered and the solvent was removed in vacuo. The residue was dissolved in acetone and filtered. After adding 60 ml of 10% potassium sulfate perfluorobutane solution dissolved in acetone, 4.7 g of crude product crystallized.

This crude product was purified by chromatography on HP-20 and 100-200 mesh to obtain 30 g of the target compound having a melting point of 235 ° C.

Example 22

(3S) -α-[[(2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] benzeneacetic acid, dipotassium salt

Dissolve 100 mg of (3S) -α-[[(2-oxo-1-sulfo-3-azetidinyl) amino] carbonyl] benzeneacetic acid, phenylmethyl ester, potassium salt (see Example 5) in 20 ml of anhydrous methanol. I was. To this was added 10% palladium (10 mg) based on activated carbon and the mixture was hydrogenated for 15 minutes. The catalyst was filtered off and methanol was evaporated in vacuo. The residue was dissolved in 5 ml of water and the pH was adjusted to 6 with 1N potassium hydroxide. The crude product obtained after freeze drying was chromatographed with HP20 resin (water was used as the eluent) to obtain 60 mg of the target compound having a melting point of 80 ° to 85 ° C.

Example 23

(S) -3- (acetylamino) -2-oxo-1-azetidinesulfonic acid, potassium salt

Method I

A) (S) -3-amino-2-oxo-1-azetidinesulfonic acid, potassium salt

169 mg of (S) -3- (benzyloxycarbonylamino) -2-oxo-1-azetidinesulfonic acid, potassium salt (see Example 3) was dissolved in 4.0 ml of water and 1 hour in the presence of 37 mg of 10% palladium based on activated carbon Hydrogenation for 40 minutes. The catalyst was filtered off and washed with 1 ml of 50% acetone aqueous solution.

B) (S) -3- (acetylamino) -2-oxo-1-azetidinesulfonic acid, potassium salt

The solution of free amine was diluted with 3.5 ml of acetone, which was stirred in an ice bath. Acetyl chloride (320 μl) was added in small portions over about 15 minutes, and the pH was adjusted to 6.5-7.2 by addition of potassium bicarbonate solid. After 30 minutes, silica gel thin layer chromatography (TLC) and aridone test in acetone: acetic acid (19: 1) showed that the reaction was largely terminated. 6 ml of 0.5M monobasic potassium phosphate buffer (pH5.5) was added and the solution was acidified to pH4.8 with 2N hydrochloric acid. Acetone was removed in vacuo, and the resulting aqueous solution was passed through a 50 ml HP-20AG column to obtain 2.197 g of a solid. This solid was dissolved in methanol to give 282 mg of extract, which still contained some salt. This product was further purified by passing through an IRC-50 column, then acidified to pH 3.8 and dried in vacuo. Titration with acetone afforded 64 mg of the target compound containing about 0.5 equivalents of inorganic potassium salt. Finally, it was passed through a 200 ml HP-20AG column, followed by freeze drying with 0.5 ml of water and drying for 2 hours at 50 ° C. in vacuo to yield 22 mg of amorphous powdery product. After softening at 100 degreeC, it became melting | fusing point 170 degrees-180 degreeC.

Analysis of C ₅ H ₇ O ₅ N ₂ SK

Theoretic value: C 24.38; H 2.87; Found: C 26.06; H 3.14;

N 11.37; K 15.90 N 9.96; K 18.04

Method II

A 2.0 g solution of (S) -2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and pyridine salt (see Example 2) dissolved in 25 ml of water was 10% based on activated carbon. Hydrogenated to 500 mg of palladium. After 2 hours, the solution was filtered, cooled to 0 ° C., and 40 ml of acetone were added. Acetyl chloride and cooled 10% potassium bicarbonate solution were added simultaneously to maintain the pH of the solution at 5.2-5.8. Acetyl chloride was added to adjust the pH of the solution to 4.2, and the solution was concentrated on a rotary evaporator to remove acetone. Chromatography was carried out on a 300 ml HP-20AG column (25 ml fraction of water eluting agent) to obtain 900 mg of the target compound containing some potassium acetate in fractions 13 and 14. Rechromatography was carried out with HP-20AG to obtain an analytical compound having a melting point of 205 ° to 210 ° C.

Analysis of C ₅ H ₇ N ₂ O ₅ SK

Theoretic value: C 24.38; H 2.86; Found: C 24.23; H 2.81;

N 11.38; S 13.02; N 11.25; S 12.86;

K 15.88 K 15.74

Example 24

(S) -2-oxo-3-[(phenoxyacetyl) amino] -1-azetidinesulfonic acid, potassium salt

1.5 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example) in 100 ml of anhydrous dimethylformamide was cooled to 0 ° C. After 2 ml of propylene oxide was added, 1 g of phenoxyacetyl chloride was added dropwise with stirring. This reaction was completed within 1 hour. The solvent was removed in vacuo and the residue was treated with 1 equivalent of potassium sulfate pifluorobutane in 20 ml of acetone. After adding ether thereto, the target compound (1 g) was crystallized, which was filtered and dried. After recrystallization in boiling water, the melting point of the target compound reached 176 ° -180 ° C.

Example 25

[3S (R ^* )]-3-[[[[[3-[(2- (furanylmethylene) amino] -2-oxo-1-imidazolidinyl] carbonyl] aminophenylacetyl] amino]- 2-oxo-1-azetidinesulfonic acid, potassium salt

(S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt in 100 ml of dimethylformamide in the presence of 1.5 g of activated carbon based palladium (see Example 4) ) 3 g were hydrogenated. After 0.5 hour, the catalyst was filtered off, and 1.8 g of dicyclohexylcarbodiimide and (R) -α-[[[3- (2-furanylmethylene) amino] -2-oxo-1-imide were added thereto. 2 g of dazolidinyl] carbonyl] amino] benzeneacetic acid and 0.9 g of hydroxybenzotriazole were added. After 3 hours, the reaction mixture was evaporated to dryness, dissolved in 50 ml of anhydrous acetone, and the precipitated urea was filtered off. After adding 1 equivalent of potassium sulfate perfluorobutane in 20 ml of acetone, the target compound precipitated. 200 ml of ether was added thereto to crystallize. It recrystallized in water after filtration, and 2g of target compounds of melting | fusing point 220 degrees-225 degreeC (decomposition) were obtained.

Example 26

[3S (R ^* )]-2-oxo-3-[[[[(2-oxo-1-imidazolidinyl) -carbonyl] amino] phenylacetyl] amino] -1-azetidinesulfonic acid, potassium salt

(S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and tetrabutylammonium salt in 100 ml of anhydrous dimethylformamide in the presence of 1.5 g of activated carbon based palladium (Example 4 3 g was hydrogenated. After 30 minutes, the catalyst was filtered off, 1.8 g of (R) -α-[[(2-oxo-1-imidazolidinyl) carbonyl] amino] benzeneacetic acid, 1.3 g of dicyclohexylcarbodiimide, and 0.9 g of hydroxybenzotriazole was added and the solution was then stirred for 2.5 hours. The solvent was removed in vacuo and the residue was dissolved in 50 ml of acetone. The precipitated urea was filtered off and the mother liquor was treated with 1 equivalent of potassium sulfate perfluorobutane in 20 ml of acetone. The desired compound was crystallized, and 200 ml of ether was added thereto, followed by filtration and recrystallization with water / acetone to obtain 1.8 g of a product having a melting point of 210 ° to 215 ° C.

Example 27

[3S (Z)]-3-[[((methoxyimino) phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

A) [3S (Z)]-3-[[(methoxyimino) phenylacetyl] amino] -2-azetidinone

After dissolving 3.58 g of (Z) -α- (methoxyimino) benzeneacetic acid in methylene chloride, it was cooled to 5 ° C and treated with a solution of 4.53 g of dicyclohexylcarbodiimide dissolved in 50 ml of methylene chloride. The mixture was stirred at 5 ° C. for 30 minutes, and thereto was added a solution of 1.72 g of 3-amino-2-azetidinone (see Example 5A) dissolved in 100 ml of methylene chloride. The reaction mixture was left at 5 ° C. for 1 hour and then at room temperature for 2 hours. Chlohexylurea was again removed by filtration, and the filtrate was evaporated to give 6.6 g of crude product. This material was purified by chromatography on 750 g of silica gel using a methylene chloride / ethyl acetate (7: 3) mixture as the eluent, to obtain 2.9 g of the product.

B) [3S (Z)]-3-[[(methoxyimino) phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

0.5 ml of pyridine was dissolved in 5 ml of anhydrous methylene chloride, then cooled to -30 DEG C, and a solution of 0.93 ml of trimethylsilyl chlorosulfonate dissolved in 5 ml of methylene chloride was added thereto. The mixture was stirred at rt for 30 min and evaporated to dryness in vacuo. The residue was dissolved in 10 ml of dimethylformamide, to which was added a solution of 1.23 g of azetidinone dissolved in 10 ml of dimethylformamide at room temperature. After stirring for 2 hours at room temperature, the solution was evaporated to dryness to give crude [3S (Z)]-3-[[(methoxyimino) phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, pyridine salt 2.1 g was obtained.

This pyridine salt was treated with tetrabutylammonium hydrogen sulfate to obtain the corresponding terabutylammonium salt, which was extracted with methylene chloride and evaporated to obtain an oily substance.

This terabutylammonium salt was treated with acetone in 1 equivalent of potassium sulfate fluorobutane, and after evaporation, the residue was treated with ether to obtain 1.6 g of potassium salt of the target compound. This was purified by HP-20 chromatography. It eluted with water / acetone (90:10) and the product of melting | fusing point 220 degreeC (decomposition) was obtained.

Example 28

[3S (Z) -3-[[(2-amino-4-thiazolyl) [[(2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] imino] acetyl] amino ] -2-oxo-1-azetidinesulfonic acid, potassium salt (1: 1)

(S) -3-amino-2-oxo-1-azetidinesulfonic acid, terabutylammonium salt (see Example 6A) dissolved in 60 ml of dimethylformamide, 0.005 mol, and (Z) -2-amino-α- [ A solution of 0.006 mol of [2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] imino] -4-thiazole acetic acid was added to 0.7 g of hydroxybenzotriazole and dicyclohexylcarbodiimide. Treated with 1.13 g. The mixture was stirred at rt for about 16 h, filtered and the filtrate was evaporated. The residue was dissolved in 30 ml of acetone, filtered and treated with 20 ml of 10% potassium sulfate perfluorobutane solution dissolved in acetone. After the addition of petroleum ether, the desired compound precipitated, which was treated with ether and filtered to give 3.8 g of product having a melting point of 190 ° C. (decomposition).

Example 29

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy] imino] acetyl] amino] -2-oxo-1-azetidinesulfonic acid Dipotassium salt

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] imino] acetyl] amino ] 2 g of oxo-1-azetidinesulfonic acid and potassium salt (see Example 28) were suspended in 5 ml of anisol, and 25 ml of trifluorotoacetic acid was added at -10 ° C. The mixture was stirred at −10 ° C. for 10 minutes, to which ether (100 ml) was slowly added at −10 ° C. followed by 50 ml of petroleum ether. The precipitate was filtered to give 1.6 g of trifluoroacetate, which was suspended in 20 ml of water at 0 ° C., and then diluted potassium hydroxide was added to adjust the pH to 5.5 and purified by HP-20 column. It eluted with water and obtained the target compound of melting | fusing point 225 degreeC (decomposition).

Example 30

[3S (Z) -3-[[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -2-oxoethoxy] imino] acetyl] amino] -2-oxo-1 Azetidinesulfonic acid, potassium salt

(Z) -2-amino-α-[[2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] imino] -4-thiazole acetic acid instead of (Z) -2- The procedure of Example 28 was repeated except that amino-α-[[2- (diphenylmethoxy) -2-oxoethoxy] imino] -4-thiazole acetic acid was used. The desired compound was obtained.

Example 31

[3S (±)]-3-[[((azidophenylacetyl) amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

Method I

A) (±, S) -α-azido-N- (2-oxo-3-azetidinyl) benzeneacetamide

2.15 g of (S) -3-amino-2-azetone (see Example 5A) and 2.1 g of sodium bicarbonate were dissolved in 50 ml of acetone / water (2: 1). 5 g of (±) -α-azidobenzeneacetyl chloride dissolved in 10 ml of acetone was added dropwise thereto, and the pH was adjusted to 6.8 by adding sodium bicarbonate while maintaining the temperature at 0 ° to 5 ° C. After stirring for 1 hour, acetone was distilled off and sodium carbonate was added to the remaining aqueous solution to adjust the pH of the aqueous solution to 8, and then extracted three times with 50 ml of methylene chloride. The organic layer was dried over sodium sulfate, titrated with ether and crystallized to obtain 3.6 g of an oily target compound. It recrystallized with methylene chloride / ether, and the target compound of 97 degreeC-100 degreeC of melting | fusing point was obtained.

B) [3S (±) [-3-[(azidophenylacetyl) amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

A solution of 2.45 g of azetidinone and 3 g of monosilyl trifluoroacetamide dissolved in 20 ml of acetonitrile was left at 40 ° C. for 1 hour, then cooled to 0 ° C., and the solution was trimethylsilyl chlorosulfonate 1.88. Treated with g and then stirred for 5 hours in the presence of argon. Finally, 6.12 ml of a 2N potassium-2-ethyl hexanoate solution in n-butanol was added and stirred for 45 minutes. This solution was poured into 300 ml of ether and the precipitate was filtered off. The solution obtained by filtering 1.2 g of precipitate in phosphate buffer (pH5.5) was chromatographed with 100 ml of HP-20. It was eluted with 20 ml of buffer, 200 ml of H ₂ O, 200 ml of water: acetone (9: 1), and 200 ml of water: acetone (3: 1). Elution was checked by TLC (SiO ₂ redon test). 25 ml fractions were taken and 280 mg of the desired compound were obtained from fractions 15 and 16. Secondary column chromatography was performed on this material to obtain 130 mg of the target compound having a melting point of 148 ° C. (decomposition).

Method II

2.03 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutyl ammonium salt (see Example 6A) and 0.9 g of (±) -α-azido-benzeneacetic acid were dissolved in 30 ml of acetonitrile. To this was added a solution of 1.03 g of dicyclohexylcarbodiimide dissolved in 10 ml of acetonitrile. It was left for 1 hour at 0 ° C. and 10 hours at room temperature. The resulting precipitate was filtered off and then the solvent was removed in vacuo and the oily residue was dissolved in 20 ml of acetone, which was then treated with 1.70 g of potassium sulfate perfluorobutane in acetone. 10 ml of ether was added thereto to crystallize the target compound at melting point of 149 DEG C (decomposition).

Method III

2.5 g of α-azidophenylacetyl chloride was added to a solution of 4.06 g of 3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) and 5 g of propylene oxide dissolved in 30 ml of acetonitrile. . After stirring for 2 hours the solvent was distilled in vacuo and the oily residue was treated with 1 equivalent of potassium sulfate perfluorobutane in acetone. After the ether was added, the desired compound crystallized out, which was filtered off. Melting Point 148 ℃ ~ 149 ℃ (Decomposition)

Example 32

[3S (D)]-3-[[[[(4-methoxyphenyl) methoxy] carbonyl] amino] phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A), dissolved in 50 ml of acetonitrile, 2.03 g and D-α-[[[(4-methoxyphenyl) methoxy] car A solution of 1.03 g of dicyclohexylcarbodiimide dissolved in 10 ml of acetonitrile was added dropwise to a solution of 1.58 g of carbonyl] amino] benzeneacetic acid. The temperature was kept at 5 ° C. for 1 hour at room temperature for 6 hours. The resulting dicyclohexylurea was isolated and then the solvent was distilled off to give the desired compound in the form of an oily residue. This oily substance was treated with potassium sulfate perfluorobutane and ether in acetone to obtain 2.4 g of a product having a melting point of 108 ° C to 111 ° C (decomposition).

Examples 33 to 37

(D) -α-[[[(4-methoxyphenyl) methoxycarbonyl] amino] benzeneacetic acid was carried out in accordance with the method of Example 32, except that the compounds described in Column I below each corresponded to the following columns The compound described in II was obtained.

Example 38

(3S) -3-[[[[(methylthio) thioxomethyl] thio] phenylacetyl] amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

1.03 g of (±) -α-[[(methylthio) thioxomethyl] thio] benzeneacetic acid and 3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutyl ammonium salt dissolved in 30 ml of acetonitrile (Example 6A) A solution of 1.63 g was treated with a solution of 0.8 g of dicyclotolsilcarbodiimide dissolved in 10 ml of acetonitrile at -5 ° C. After stirring for about 16 hours, the resulting dicyclohexylurea was filtered off and the mother liquor was evaporated. The obtained oily residue was chromatographed with a column of 400 g of silica, to obtain 1.3 g of the target compound [eluent = ethyl acetate / methanol / water (8.5: 1: 0.5)].

Example 39

3 (S) -3-[[[[(methylthio) thioxomethyl] thio] phenylacetyl] amino-2-oxo-1-azetidinesulfonic acid, potassium salt

1.3 g of (3S) -3-[[[[(methylthio) thioxomethyl] thio] phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 38) was added to acetone. It was dissolved and treated with 1 equivalent of potassium sulfate pifluorobutane. The target compound crystallized by adding ether was filtered to obtain 0.18 g of a product having a melting point of 157 占 폚 (decomposition).

Example 40

[3S (D)]-3-[[[[(4-ethyl-2,3-dioxo-1-piperazinyl) -carbonyl] amino] -2-thienylacetyl] amino] -2-oxo -1-azetidinesulfonic acid, potassium salt

(D) -α-[[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] -2-thiophenacetic acid 3.25 g, 3-amino-2-oxo-1- 4.20 g of azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) and 1.3 g of N-hydroxybenzotriazole were dissolved in 25 ml of acetonitrile. 2.06 g of dicyclo hexylcarbodiimide dissolved in 10 ml of acetonitrile was added dropwise at -10 ° C for 20 minutes and stirred for about 16 hours. The resulting urea was filtered off, the solvent was evaporated in vacuo, and the remaining oil was dissolved in 50 ml of acetone, which was then treated with 1 equivalent of potassium sulfate pifluorobutane to determine a target compound having a melting point of 185 ° to 187 ° C. (decomposition). Got as.

Example 41

[3S (±)]-3-[(bromophenylacetyl) amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

A solution of 1.4 g of α-bromophenylacetyl chloride dissolved in 10 ml of acetonitrile was dissolved in a solution of 5 mM 3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt and 3 g of propylene oxide in acetonitrile at 0 ° C. Added dropwise. After stirring for 3 hours, the solvent was distilled off and the remaining oily residue was dissolved in 30 ml of acetone. After adding 1 equivalent of potassium sulfate pifluorobutane dissolved in acetone to this, ether was added, and the target compound of melting | fusing point 135 degreeC-137 degreeC (decomposition) crystallized.

Example 42

[3S (±)]-3-[[[(aminocarbonyl) amino] -2-thienyl acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

Method I

(±) -2-amino-4- (2-thienyl) in a solution of (S) -3-amino-2-oxo-1-azetidinesulfonic acid, 2 g of tetrabutylammonium salt and 1.5 g of propylene oxide dissolved in acetonitrile 1.1 g of -5 (4H) -oxazolone, hydrochloride was added and stirred at 0 ° C. for 3 hours and at room temperature for 1 hour, after which the solvent was distilled off and the oily residue was dissolved in 30 ml of acetone. 1 equivalent of potassium sulfate pifluorobutane dissolved in acetone was added to determine the target compound, which was subjected to column chromatography with HP-20 using water as the eluent, to a melting point of 218 ° to 222 ° C. (decomposition). The compound was obtained.

Method II

2 g of (±) -α-[(aminocarbonyl) amino] -2-thiophenacetic acid suspended in 10 ml of acetonitrile, (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt 10 mM And 10 mM dicyclohexylcarbodiimide dissolved in 15 ml of acetonitrile at 0 ° C. in a suspension of 10 mM of N-hydroxybenzotriazole was added while stirring. After stirring at −15 ° C. for 1 hour and at room temperature for about 16 hours, the dicyclohexyl urea was filtered off and the solvent was evaporated to dissolve the resulting oily residue in 50 ml of acetone. After adding 1 equivalent of potassium sulfate pifluorobutane dissolved in acetone to it, a crystalline product was obtained. Column chromatography with HP-20 was carried out using water as the eluent to afford the purified desired compound, which had a melting point of 220 ° to 223 ° C.

Example 43

[3S (±)]-3-[[[((methylamino) carbonyl] amino] -2-thienylacetyl] amino] -2-oxo-1-azetidinesulfonic acid potassium salt

0.54 g of (±) -α [[(methylamino) carbonyl] amino] -2-thiophenacetic acid and (S) -3-amino-2-oxo-1-azetidinesulfonic acid tetrabutylammonium salt (see Example 6A 1.00 g) was dissolved in 20 ml of acetonitrile, to which 0.5 g of dicyclohexylcarbodiimide was added at 0 ° C. After stirring for 8 hours, dicyclohexylurea was filtered off, the solvent was distilled off, and the resulting oily residue was dissolved in 50 ml of acetone, to which 1 equivalent of potassium sulfate pifluorobutane was added, and The crystalline product thus obtained was filtered and then purified by HP-20 using water as eluent. The melting point of this product was 205 ° C. (decomposition).

Example 44

[3S (±)]-3-[[[(aminooxoacetyl) amino] -2-thienylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

Example 42, method except using (±) -α [(aminooxoacetyl) amino] -2-thiophenacetic acid instead of (±) -α [(aminocarbonyl) amino] -2-thiophenacetic acid It carried out according to the process of II, and obtained the target compound of melting | fusing point 218 degreeC-222 degreeC.

Example 45

[3S (R ^* )]-3-[[[(4-ethyl-2,3-dioxo-1-piperazinyl) -carbonyl] amino] phenylacetyl] amino-2-oxo-1-azetidine Sulfonic acid, potassium salt

(D) -α-[[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] -amino] -2-thiophenacetic acid instead of (R) -α-[[(4 Except for using -ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] benzeneacetic acid, it carried out in accordance with the method of Example 40, and the target compound of melting | fusing point 155 degreeC-157 degreeC (decomposition) was obtained. Got it.

Example 46

3- (acetylamino) -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

[Method I]

A) Sodium and potassium mixed salts of 3-[(N-acetyl-N-chloro) amino] -2-oxo-1-azetidinesulfonic acid

172 mg of 3- (acetylamino) -2-oxo-1-azetidinesulfonic acid, potassium salt (see Example 23) dissolved in 17 ml of methanol containing 4% sodium borate decahydrate cooled to -15 ° C to -10 ° C. 110 μl of t-butyl hypochlorite was added to the solution. The mixture was stirred for 1 h and 45 min in a cold place, poured into 50 ml of 0.5 M monobasic potassium phosphate solution, and the pH was lowered to 5.5. The solvent was removed in vacuo and the residue was dissolved in a minimum amount of water and then chromatographed with HP-20AG, 100-200 mesh (140 ml). Elution with water gave 63 mg of an oily substance, which was left to crystallize. This was titrated with methanol / ether and then with ether to obtain 53 mg of a solid having a melting point of 124 ° C (degrading slowly).

B) 3- (acetylamino) -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

37 mg of 3-[(N-acetyl-N-chloro) amino] -2-oxo-1-azetidinesulfonic acid mixed salt was dissolved in 1.5 ml of anhydrous dimethylformamide, and 50 ml of lithium methoxide dissolved in 1 ml of methanol. Solution was added at -78 ° C. After stirring for 15 minutes at −78 ° C., 10 ml of a 0.5 M monobasic potassium phosphate solution was added thereto, and 1N hydrochloric acid was added to the solution to acidify to pH 4.

After adding 70 mg of hydrogen tetrabutylammonium sulfate to this solution, this solution was extracted 4 times with methylene chloride. The extracts were combined, dried over sodium sulfate, and the solvent was removed in vacuo to give 55 mg of an oily substance. Chromatography was performed with 5.5 g of silica gel (eluent = 8% methanol: 92% methylene chloride) to obtain 41 mg of an oily substance in the form of tetrabutylammonium salt. 31 mg of this oily substance was dissolved in water, passed through an ion exchange column (AG50W-X2, 5ml, K ⁺ type, 200-400 mesh, 0.6meq / ml), and the oily substance was removed by vacuum removal. Crystallized with. Titration twice with ethylene gave 11 mg of a colorless powder product having a melting point of 182 ° to 183 ° C (decomposition).

[Method II]

A) 3-Amino-3-methoxy-2-oxo-1-azetininsulfonic acid, tetrabutylammonium salt

100 µl of a 4% sodium borate decahydrate solution dissolved in methanol was added to a suspension of 30 mg of activated carbon based 10% palladium suspended in 2 ml of methanol, and the mixture was stirred for 15 minutes under a hydrogen atmosphere. To this was added 60 mg of 3-methoxy-2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 49) in 2 ml of methanol. The mixture was stirred vigorously for 15 minutes under hydrogen atmosphere. The catalyst was removed by filtration with celite (0.5 mμ) of microfiltration paper, the solvent was removed from the filtrate in vacuo, and the residue was extracted with methylene chloride. The solvent was removed under reduced pressure to obtain 35 mg of an oily target compound.

B) 3- (acetylamino) -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

To a solution of 35 mg of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid and tetrabutyl ammonium salt dissolved in 10 ml of methylene chloride, 2 ml of propylene oxide and 74 µl of acetyl chloride were added at 0 ° C. After 2 hours, the solvent was removed under reduced pressure, and the remaining oily substance was chromatographed with 4 g of silica gel. Elution with 6-8% methanol in methylene chloride gave 18 mg of an oily substance, which was dissolved in water and passed through an ion exchange resin (AG50-X2 3 ml, K ⁺ type, 0.6 meq / ml). Water was removed from the eluate in vacuo to afford 10 mg of the target compound.

Example 47

N- (3-methoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide, tetrabutylammonium salt

A) N-chloro-N- (2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide potassium salt

(S) -N- (2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide, potassium dissolved in methanol containing 5 ml of 4% sodium borate decahydrate cooled in a -5 ° C bath To 50 mg of the salt (see Example 1) was added 20 μl of t-butylhypochlorite. After stirring for 32 minutes, the mixture was poured into 0.5 M potassium phosphate buffer (pH 5.5) at 0 ° C. The pH of the resulting solution (pH5.9) was adjusted to 4.5, concentrated in vacuo to remove methanol, and then chromatographed with HP-20AG, 100-200 mesh (100 ml). The column was washed with 100 ml of 0.5 M buffer (pH 5.5) and water, and the desired product was eluted with water: acetone (9: 1). This was concentrated in vacuo to yield 50 mg of the target compound.

B) N- (3-methoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide, tetrabutylammonium salt

N-chloro- (2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetate dissolved in 10 ml of anhydrous dimethylformamide in a solution of 160 mg of lithium methoxide dissolved in 5 ml of methanol cooled to -78 ° C. A solution of 149 mg of amide, potassium salt was added. After complete addition, the mixture was stirred at −78 ° C. for 15 minutes, poured into 100 ml of 0.5N monobasic potassium phosphate solution, and washed three times with methylene chloride. 213 mg of tetrabutylammonium disulfate was added to the aqueous layer, which was then extracted three times with methylene chloride. The combined extracts were dried over sodium sulfate and the solvent was removed in vacuo to give 271 mg of an oily substance. Chromatography with 25 g of silica gel eluted with 4% methanol: 96% methylene chloride to obtain 149 mg of an oily product.

Example 48

N- (3-methoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenylacetamide, potassium salt

91 mg of N- (3-methoxy-2-oxo-1-sulfo-3-azetidinylphenyl) -2-phenylacetamide, tetrabutylammonium salt (see Example 47) were dissolved in water, and an ion exchange column ( 10 ml of AG 50W-X2, type K ⁺ ). The eluate was concentrated in vacuo and the resulting oily residue was solidified by scratching with a methanol-acetone-ether mixture. After titration twice with ether, 53 mg of solid product was obtained.

Example 49

3-methoxy-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt

[Method I]

A) 2-oxo-3- [N-chloro-N-[(phenylmethoxy) carbonyl] -amino] -1-azetidinesulfonic acid, tetrabutylammonium salt

0.9 g of (S) -2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 4) dissolved in 80 ml of methylene chloride was dissolved in sodium borate 10 To a mixture (cool to 0-5 ° C.) of 3.17 g of hydrate and 11.8 ml of a 5.25% sodium hypochloride solution in 70 ml of water, the reaction mixture was stirred vigorously for 55 minutes while cooling in an ice bath. The mixture was diluted with 0.5 M monobasic potassium phosphate solution, and the product was extracted three times with 150 ml of methylene chloride. The extracts were combined, dried over sodium sulfate, and the solvent was removed in vacuo to obtain 0.94 g of an oily target compound.

B) 3-methoxy-2-oxo-3-[[(phenylmethoxy) carbonyl] -amino] -1-azetidinesulfonic acid, tetrabutylammonium salt

2-oxo-3- [N-chloro-N-[(phenylmethoxy) carbonyl] amino] -1- dissolved in 10 ml of anhydrous dimethylformamide in a high solution of 667 mg of lithium methoxide dissolved in 10 ml of anhydrous methanol. A solution of azetidinesulfonic acid, 0.94 g of tetrabutylammonium salt was added. The mixture was stirred at −78 ° C. for 1 hour, poured into a 0.5 M monobasic potassium phosphate solution, and extracted three times with 150 ml of methylene chloride. The extracts were combined and dried over sodium sulfate, and the solvent was removed in vacuo to give 0.83 g of an oily substance. This oily substance was chromatographed with 100 g of silica gel and eluted with 4-5% methanol in methylene chloride to obtain 513 mg of the target compound in the oily form.

[Method II]

In a solution of 400 mg of 3-benzyloxycarbonylamino-3-methoxy-2-oxo-1-azetidinesulfonic acid and potassium salt dissolved in water, 0.1M tetrabutylammonium disulfide solution (potassium hydroxide to pH 4.3 Control) 10.9 ml are added and the mixture is extracted three times with methylene chloride, and the extracts are combined and dried over Na ₂ SO ₄ , and the solvent is removed in vacuo to give similar spectral properties as the product of Method I. 625 mg of phase material was obtained.

Example 50

3-methoxy-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, calcare salt

[Method I]

A) 2-oxo-3- [N-chloro-N-] (phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

(S) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt dissolved in 90 ml of methanol containing 4% sodium borate decahydrate (see Example 3) 420 g of t-butyl hypochlorite were added at -10 ° C. After stirring for 2 hours at -10 DEG C, 100 ml of 0.5 M monobasic potassium phosphate solution was added thereto, followed by addition of 1N hydrochloric acid to adjust the pH to 6. The solvent was concentrated to 30 ml in vacuo and then chromatographed with 200 ml of HP-20AG, 100-200 mesh. A solution of 50 g of monobasic potassium phosphate dissolved in 1,000 ml of water was passed through, followed by 2,000 ml of water, and the product was eluted with 10% acetone to 90% water. The solvent was removed in vacuo and crystallized with water to give 530 mg of solid target compound having a melting point of 173 ° to 175 ° C.

B) 3-methoxy-2-oxo-3-[[(phenyl, methoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

In a solution of 874 mg of lithium methoxide dissolved in 10 ml of anhydrous methanol, 2-oxo-3- [N-chloro-N-[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid in 13 ml of anhydrous dimethylformamide, 857 mg potassium salt was added at -78 ° C. After holding this mixture at -78 ° C for 15 minutes, the mixture was poured into 200 ml of 0.5M monobasic potassium phosphate solution, and 1N hydrochloric acid was added thereto to adjust the pH to 5.5. The aqueous mixture was washed three times with 100 ml each of methylene chloride, and 1.169 g of tetrabutylammonium disulfide was added thereto. The product was extracted three times with 200 ml each of methylene chloride, dried over sodium sulfate, filtered and concentrated in vacuo. The residual oily substance was chromatographed with 150 g of silica gel, and the product was eluted with 2-4% methanol in methylene chloride to give 701 mg of tetrabutylammonium salt of the product as an oil phase. 51 mg of this oily substance was dissolved in water and passed through an ion exchange column (3 ml of AG 50W-X2, 200-400 mesh, K ⁺ type, 0.6 meq / ml). 30 mg of the oily substance produced by concentrating the eluate in vacuo was crystallized by scratching with acetone (melting point 196 ° -198 ° C.).

[Method II]

A) 1-chloro-3- [N-chloro-N-[(phenylmethoxy)] carbonyl-2-azetidinone

A solution of 440 mg of 3-[[((phenylmethoxy) carbonyl] amino] -2-azetidinone (see Example 2C) dissolved in 40 ml of 4% methanolic borax (see Example 2C) was cooled to 0 ° C. and t- 0.5 ml of butylhypochlorite was added. After holding at 0 ° C. for 30 minutes, the solution was poured into 200 ml of cold water, and extracted twice with 100 ml of ethyl acetate. The ethyl acetate layers were combined, washed with water, dried and evaporated in vacuo to afford 546 mg of the oily target compound.

B) 3-methoxy-3-[[phenylmethoxy) carbonyl] amino] -2-azetidinone

A solution of 730 mg (0.0025 mol) of 1-chloro-3- [N-chloro-N-[(phenylmethoxy) carbonyl] amino] -2-azetidinone dissolved in 5 ml of tetrahydrofuran was cooled to -78 ° C. To this was added 4 ml of methanol containing 285 mg of lithium methoxide. After 20 minutes at −78 ° C., 0.6 ml of acetic acid and 0.6 ml of trimethylphosphite were added thereto. The solution was stirred at −78 ° C. for 5 minutes, warmed to ambient temperature and then stirred for 30 minutes. The resulting solution was diluted with ethyl acetate and then washed with 5% sodium bicarbonate, water, 5% potassium disulfate, water, saturated salt solution and dehydrated. The oily material obtained by removing the solvent was developed with ethyl benzene-ethyl acetate (1: 1) using four 20 x 20 cm x 1 mm silica gel plates, and then the main UV-active band portion having an Rf value of 0.25 was isolated. 91 mg were obtained, which was crystallized with ether to give a solid. This was recrystallized with ether to obtain the target compound having a melting point of 112 ° to 114 ° C.

C) 3-methoxy-2-oxo-3-[[(phenylmethoxy) carbonyl] -amino] -1-azetidinesulfonic acid, potassium salt

55.4 mg of pyridine-sulfur trioxide complex was added to a solution of 25 mg of 3-methoxy-3-[[(phenylmethoxy) carbonyl] amino] -2-azetidinone dissolved in 0.175 ml of dichloromethane and dimethylformamide, respectively. And stirred for 24 hours. The resulting slurry was diluted with 5 ml of chilled 0.5 M monobasic potassium phosphate (adjusted to pH 4.5) and then extracted with ethyl acetate. The aqueous layer was eluted with buffer, water and water-acetone (9: 1) using a 40 ml HP-20AG column to afford the target compound in the oil phase, which slowly solidified. Crystallization with acetone afforded the target compound with a melting point of 196 ° to 198 ° C (decomposition).

Example 51

3-[[1,3-dioxo-2-phenyl-3- (phenylmethoxy) propyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

A) 3-[[1,3-dioxo-2-phenyl-3- (phenylmethoxy) propyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

431 mg of crude 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 74) containing borax was dissolved in 30 ml of anhydrous acetonitrile. 317 μl of anhydrous pyridine was added and the solution was stirred well at −10 ° C. under a dry nitrogen atmosphere, and then 568 mg of α- (benzyloxycarbonyl) phenyl acetyl chloride dissolved in 3 ml of anhydrous acetonitrile was added dropwise. The reaction was complete after 15 minutes by thin layer chromatography. To this was added 17 ml of 0.5 M potassium phosphate buffer (pH 5.5) and most of the acetonitrile was removed in vacuo. The residue was diluted with water and extracted three times with the same volume of methylene chloride. The extract was dried over anhydrous sodium sulfate and evaporated in vacuo to afford 1.032 g of crude product. This was dissolved in 3 ml of methylene chloride and subjected to silica gel chromatography using methylene chloride-methanol as the eluent to obtain 470 mg of the target compound.

B) 3-[[1,3-dioxo-2-phenyl-3- (phenylmethoxy) -propyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

30% of 470 mg of 3-[[1,3-dioxo-2-phenyl-3- (phenylmethoxy) -propyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt It was dissolved in 15 ml of acetone-water and passed through a Dowex 50W-X2 (K ⁺ type) column using the same solvent as eluent. All eluates were evaporated in vacuo to give 345 mg of an amorphous solid, which was freeze-dried to obtain an amorphous powder having a melting point of 100 ° to 120 ° C.

Analysis of C ₂₀ H ₁₉ O ₈ N ₂ SK

Theoretic value: C 49.37; H 3.94; Found: C 49.08; H 4.00;

N 5.76; S 6.59 N 5.58; S 6.29

Example 52

(±) -3-[[[((cyanomethyl) thio)] acetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

To a solution of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 74), 414 mg dissolved in 50 ml of anhydrous acetonitrile (210), 210 µl of diethylaniline and- 169 mg of nomethylthioacetyl chloride was added. After 10 minutes, the solvent was removed under reduced pressure, thereto was added potassium hydroxide to adjust the pH of 0.1M tetrabutyl ammonium sulfate (22.8 ml) to 4.3, and the obtained product was extracted three times with 50 ml of methylene chloride, and dried, Filtered and then concentrated under reduced pressure. The oily residue was purified with 60 g of silica gel and 294 mg of the product was eluted with 4% methanone-methylene chloride. The purified product was passed through an ion exchange resin (AG50W-X2 (K ⁺ type) 16ml 0.6meq / ml, 200-400 mesh). Water was removed to give 164 mg of partially purified product. This product was further purified with Dianion AG HP 20 (100 ml) using water as eluent. The solvent was removed under reduced pressure to obtain 126 mg of the product, which was titrated with ether to obtain 76 mg of the target compound having a melting point of 110 ° to 125 ° C.

Analysis of C ₈ H ₁₀ N ₃ S ₂ O ₆ K

Theoretic value: C 27.66; H 2.88; Found: C 27.25; H 3.00;

N 12.10; S 18.44 N 10.84; S 17.53

Example 57

[3S (±)]-3-[[2- (methylthio) -1-oxopropyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

(±) -2- (methylthio) -propanoic acid was used in place of (±) -α-azido-benzeneacetic acid and was carried out in accordance with the method of Example 31, Method II to achieve a melting point of 172 占 폚 (decomposition). The desired compound was obtained.

Example 58

[3S (R ^* )]-3-[[[[2,3-dioxo-4-[(phenylmethylene) amino] -1-piperazinyl] carbonyl] amino] phenylacetyl] amino] -2- Oxo-1-azetidinesulfonic acid, potassium salt

0.7 g of (R) -α [[[2,3-dioxo-4-[(phenylmethylene) amino] -1-piperazinyl] carbonyl] amino] benzeneacetic acid and (S) -3-amino-2 0.8 g of oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) was dissolved in 20 ml of acetonitrile. While stirring the solution, 0.4 g of dicyclohexylcarbodiimide was added dropwise to the solution at 0 ° C, and the stirring was continued for 18 hours. The solution was filtered, the solvent was distilled off, and the resulting oily residue was dissolved in acetone and treated with potassium sulfate perfluurobutane to precipitate the desired compound. Purification by column chromatography on HP-20 using water as the eluent gave a target compound having a melting point of 193 ° to 193 ° C.

Example 59

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(2-methoxy-2-oxoethoxy) imino] acetyl] amino] -2-oxo-1-azetidine Sulfonic acid, potassium salt

1.3 g of (Z) -2-amino-α [(2-methoxy-2-oxoethoxy) imino-4-thiazole acetic acid and (S) -3-amino-2-oxo-1-azetidinesulfonic acid 2.03 g of tetrabutylammonium salt (see Example 6A) was dissolved in 50 ml of acetonitrile, and 1.03 g of dicyclohexylcarbodiimide dissolved in 5 ml of acetonitrile was added dropwise at 0 占 폚. After stirring for 15 hours, dicyclohexylurea was filtered off and the solvent was distilled off. The oily residue was dissolved in acetone and treated with 1 equivalent of potassium sulfate perfluurobutane. The target compound was isolated and purified by column chromatography with HP-20 using water as the eluent to obtain the target compound having a melting point of 195 ° -198 ° C.

Example 60

[3S (R ^* )]-3-[[[[[3-[[(4-chlorophenyl) methylene] amino] -2-oxo-1-imidazolidinyl] carbonyl] amino] phenylacetyl] amino ] -1-azetidinesulfonic acid, potassium salt

(S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutyl ammonium salt dissolved in 100 ml of anhydrous diglyme (diethylene glycol dimethyl ether) (see Example 6A) 1.5 g, (R) -α [ 1.5 g of [[3-[[4-chlorophenyl) methylene] amino] -2-oxo-1-amidazolidinyl] carbonyl] amino] benzeneacetic acid, 1 equivalent of dicyclohexyl-carbodiimide and hydroxide 0.5 g of oxybenzotriazole was stirred together for 12 hours. The solvent was removed in vacuo and the residue was dissolved in 50 ml of acetone and filtered. Acetone was distilled off and the residue was dissolved in 200 ml of methylene chloride. The solution was washed with an aqueous sodium bicarbonate solution followed by an aqueous sodium chloride solution and then the methylene chloride layer was dried over sodium sulfate, evaporated to dryness and crystallized by addition of ether. This compound was recrystallized with acetone-ether, and the resulting white crystalline powder was dissolved in acetone and treated with 1 equivalent of potassium sulfate perfluorobutane dissolved in acetone. The desired compound precipitated was filtered to obtain 1.4 g of a product having a melting point of 217 ° to 222 ° C.

Example 61

[3S (R ^* )]-3-[[[[2-oxo-3-[(phenylmethylene) amino] -1-imidazolidinyl] carbonyl] amino] phenylacetyl] amino-1-azetidinesulfonic acid Potassium salt

1 equivalent of dicyclohexylcart in a solution of 2.25 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 6A) dissolved in 100 ml of anhydrous dimethylformamide (DMF). Bodyimide, (R) -α-[[[2-oxo-3-[(phenylmethylene) amino] -1-imidazolidinyl] carbonyl] amino] benzene acetate 2.5g and hydroxybenzotriazole 0.85g Was added and stirred at ambient temperature for 12 hours. The DMF was then removed in vacuo and the residue dissolved in 50 ml of acetone. The precipitated urea was filtered off and the mother liquor was treated with 1 equivalent of potassium sulfate perfluorobutane dissolved in 20 ml of acetone. After addition of 100 ml of ether, the precipitated target compound was filtered off. This compound was dissolved in DMF / acetone and then precipitated with water to give 1.5 g of purified product. Melting point 224 ° -226 ° C. (decomposition).

Example 62

[3S (R ^* )]-3-[[[[3- (methylsulfonyl) -2-oxo-1-imidazolidinyl] carbonyl] amino] phenylacetyl] amino] -2-oxo-1- Azetidinesulfonic acid, potassium salt

To a solution of (R) -α-[[[3-, (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 6A) dissolved in 60 ml of dimethylformamide. (Methylsulfonyl) -2-oxo-1-imidazolidinyl] carbonyl] amino] 1.9 g of benzeneacetic acid, 0.75 g of hydroxybenzotriazole and 2.3 g of dicyclohexylcarbodiimide were added to the plate for 12 hours. I was. The solvent was removed in vacuo and the residue was dissolved in 20 ml of acetone and filtered. The mother liquor was treated with 1.87 g of potassium sulfate perfluorobutane dissolved in 20 ml of acetone. After the addition of ether, 2.0 g of the target compound was precipitated. This compound was recrystallized from water to obtain a compound having a melting point of 240 ° to 245 ° C (decomposition).

Example 63

[3S (R ^* )]-3-[(hydroxyphenylacetyl) amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

1.5 g of dicyclohexylcarbodiimide and 1.5 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 6A) dissolved in 100 ml of anhydrous dimethylformamide 0.5 g of hydroxybenzotriazole and 0.6 g of R-α-hydroxybenzeneacetic acid were added and stirred for about 16 hours. The solvent was removed in vacuo and the residue was dissolved in 20 ml of acetone. The precipitated urea was filtered off and the mother liquor was treated with 1 equivalent of potassium sulfate perfluorobutane. After adding ether thereto, the precipitated title compound was filtered to give 1.3 g of crude product. The product was chromatographed and purified with HP-20 using water / acetone (9: 1) as the eluent, and the melting point of this product was 145 ° -150 ° C. (decomposition).

Example 64

[3S (S ^* )]-3-[(hydroxyphenylacetyl) amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

It carried out in accordance with the method of Example 63 using (S) -α-hydroxybenzene acetic acid instead of (R) -α-hydroxybenzene acetic acid, to obtain a target compound having a melting point of 195 ° to 197 ° C.

Example 65

[3S (±)]-2-oxo-3-[(phenylsulfoacetyl) amino] -1-azetidinesulfonic acid, potassium salt (1: 2)

2.4 g of triethylamine and 0.3 g of dimethylaminopyridine in a solution of 2.25 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 6A) dissolved in 100 ml of anhydrous diglyme. Was added and cooled to 0 ° C. To this was added dropwise 1.8 g of (R) -α- (chlorocarbonyl) -benzenemethanesulfonic acid dissolved in 20 ml of diglyme, and this temperature was maintained for 2 hours. The solvent was distilled in vacuo and the residue was dissolved in acetone. The insoluble precipitate was filtered off and the mother liquor was treated with 1 equivalent of potassium sulfate perfluorobutane. After adding ether thereto, the precipitated target compound was filtered to obtain 1.4 g of crude product. After recrystallization of this product with water / acetone, the melting point of this product was 240 ° -245 ° C. (decomposition).

Example 66

[3S (Z)]-3-[[[(2-amino-4-thiazolyl) [[(diethoxyspinyl) methoxy] imino] acetyl] amino] -2-oxo-1-azetidine Sulfonic acid, potassium salt

A solution of 2.25 g of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 6A) dissolved in 100 ml of anhydrous dimethylformamide was added to (Z) -2-amino-α-. It was stirred for 12 hours with 1.87 g of [[(diethoxy phosphinyl) methoxy] imino] -4-thiazole acetic acid, 0.75 g of hydroxybenzotriazole and 2.29 g of dicyclohexylcarbodiimide. The precipitated urea was filtered off and the solvent was removed in vacuo. The oily residue was treated with 1 equivalent of potassium sulfate perfluorobutane dissolved in 20 ml of acetone. Ether was added thereto and the precipitated target compound was filtered to give 2.77 g of crude product. This product was purified by column chromatography using HP-20 using water / acetone (9: 1) as the eluent to obtain the target compound having a melting point of 155 ° C. to 150 ° C. (decomposition).

Example 67

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[2- (1, 1-dimethylethoxy) -2-oxo-1-phenylethoxy] imino] acetyl] Amino] -2-oxo-1-azetidinesulfonic acid, potassium salt (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt dissolved in 60 ml of dimethylformamide (see Example 6A) 2.25 2.4 g of (Z) -2-amino-α-[[2- (1,1-dimethylethoxy) -2-oxo-1-phenylethoxy] imino] -4-thiazole acetic acid in a solution of g, 1 g of hydroxybenzotriazole and 1.5 g of dicyclohexylcarbodiimide were added and stirred at room temperature for 12 hours. The solvent was removed in vacuo and the residue was dissolved in 50 ml of acetone. The precipitated urea was filtered off and the mother liquor was treated with 1 equivalent of potassium sulfate perfluorobutane. After adding ether thereto, the crystallized target compound was filtered out. This compound was purified by HP-20 column chromatography using water / acetone (7: 3) as the eluent to obtain 1 g of a product having a melting point of 250 ° C. or higher (decomposition).

Example 68

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(1H-tetrazol-5-ylmethoxyamino] acetyl] amino-2-oxo-1-azetidinesulfonic acid, potassium salt

To a solution of (S) -3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 6A) dissolved in 60 ml of dimethylformamide (Z) -2-amino-α- [ 1.4 g of (1H-tetrazol-5-ylmethoxy) imino] -4-thiazole acetic acid, 0.7 g of hydroxybenzotriazole, and 1.4 g of dicyclohexylcarbodiimide were added to a plate for 24 hours. After the solvent was removed in vacuo, the residue was dissolved in acetone and the precipitated urea was filtered off. The mother liquor was treated with 1 equivalent of potassium sulfate perfluorobutane dissolved in 10 ml of acetone. To this was added 200 ml of ether to precipitate the target compound, which was quenched by HP-20 column chromatography using water as eluent to obtain 1.05 g of a product having a melting point of 250 ° C. (decomposition).

Example 69

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(phenylmethoxy) imino] acetyl] amino-2-oxo-1-azetidine sulfonic acid, potassium salt

(S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutyl ammonium salt (see Example 6A) 1.5 g, (Z) -2-amino-α-[(phenylmethoxy) imino]- 1.23 g of 4-thiazole acetic acid, 0.57 g of hydroxybenzotriazole and 1.14 g of dicyclohexylcarbodiimide were stirred in 60 ml of dimethylformamide at room temperature for 24 hours. The precipitated urea was filtered off, the solvent was removed and the residue was treated with 1 equivalent of potassium sulfate perfluorobutane dissolved in 10 ml of acetone. 200 ml of ether was added thereto to precipitate the desired compound, which was then purified by HP-20 column chromatography using water / acetone (9: 1) as the eluent to obtain 1 g of a substance having a melting point of 200 ° C. (decomposition). Got.

Example 70

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(carboxymethoxy) imino] acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -2-oxoethoxy] imino] acetyl] amino] -2-oxo- 1.3 g of 1-azetidinesulfonic acid, potassium salt (1: 1) (see Example 30) was mixed with 5 ml of aniazole, and 25 ml of trifluoroacetic acid was added at -15 ° C, followed by stirring for 10 minutes. I was. To this was slowly added 100 ml of ether at −10 ° C. followed by 50 ml of petroleum ether. The precipitate was suspended in 20 ml of water while cooling and the pH of this suspension was adjusted to 5.0 by addition of dilute potassium hydroxide. The product was purified by HP-20 column chromatography to obtain 3.0 g of the target compound at a melting point of 230 ° to 235 ° C (decomposition).

Example 71

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(2-oxo-2- (phenylmethoxy) ethoxy] imino] acetyl] amino] -2-oxo-1 Azetidinesulfonic acid, potassium salt

(Z) -2- instead of (Z) -2-amino-α-[[2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] imino] -4-thiazole acetic acid Melting point about 170 ° C. (decomposition) using amino-α-[[2-oxo-2- (phenylmethoxy) ethoxy] imino] -4-thiazoleacetic acid and carried out according to the method of Example 28 The compound was obtained.

Example 72

[3S (Z)]-3-[[(2-amino-2-oxoethoxy) imino] (2-amino-4-thiazolyl) acetyl] amino] -2-oxo-1-azetidinesulfonic acid, Potassium salt

(Z) -2- instead of (Z) -2-amino-α-[[2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] imino] -4-thiazole acetic acid Amino-α-[(2-amino-2-oxoethoxy) imino] -4-thiazole acetic acid was carried out according to the method of Example 28, and the target compound having a melting point of 205 ° C to 210 ° C (decomposition) Got.

Example 73

[3S (Z)]-3-[[(2-Amino-4-thiazolyl) (hydroxyimino) -acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

10 g of 4A molecular sieve was added to a solution of 0.6 g of 90% hydroxybenzotriazole dissolved in 100 ml of dimethylformamide, stirred for 1 hour, and filtered, and the filtrate was dissolved in difetylformamide (S) -3- Amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) participated in a 0.04 mole solution. 0.89 g of (Z) -2-amino-α- (hydroxyimino) -4-thiazole acetic acid was added thereto, followed by 0.91 g of dicyclohexylcarbodiimide. The mixture was stirred for about 16 hours, and the residue obtained by evaporation in vacuo was dissolved in 20 ml of acetone and filtered. This was added to a potassium sulfate perfluorobutane solution to precipitate the target compound. This compound was chromatographed with HP-20 resin to obtain 0.44 g of a product having a melting point of 240 ° C. or higher.

Example 74

3-methoxy-2-oxo-3-[(2-thienylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

A) 3-Amino-3-methoxy-2-oxo-1-azetidinesulfonic acid tetrabutylammonium salt

143 mg of (±) -3-methoxy-3-[[((fanylmethoxy) carbonyl] amino] -2-oxo-1-azetidinesulfonic acid tetrabutylammonium salt (see Example 49) were dissolved in 15 ml of anhydrous methanol. . 12 mg (0.1 equiv) of Na ₂ B ₄ O ₇ · 10H ₂ O was added thereto, followed by 72 mg of 10% palladium based on activated carbon. The mixture was hydrogenated at 1 atmosphere for 15 minutes, filtered to remove the catalyst, and the filtrate was evaporated in vacuo to afford 114 mg of the desired compound.

B) 3-methoxy-2-oxo-3-[(2-thienylacetyl) amino-1-azetidinesulfonic acid, tetrabutylammonium salt

102 mg of 3-methoxy-3-amino-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt were dissolved in 10 ml of anhydrous acetonitrile, and 56.5 µl of pyridine anhydride was added thereto, and then -10 ° C under a dry nitrogen atmosphere. Stir well at 44 µl of thienylacetyl chloride dissolved in 1 ml of anhydrous acetonitrile was added dropwise thereto. Thin layer chromatography revealed that the reaction was completed within 15 minutes. To this was added 4.2 ml of 0.5 M potassium phosphate buffer (pH 5.5) and 8.5 mg (0.1 equiv) of tetrabutylammonium sulfate, removing most of the acetonitrile in vacuo. The residue was diluted with water and extracted three times with 20 ml of methylene chloride. The extract was dried over anhydrous sodium sulfate and evaporated in vacuo to give 107 mg of gummy material. The obtained crude product was purified by silica gel chromatography using methylene chloride-methanol as the eluent to obtain 66 mg of the target compound.

C) 3-methoxy-2-oxo-3-[(2-thienylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

After dissolving 154 mg of 3-methoxy-2-oxo-3-[(2-thienylacetyl) amino] -1-azetidinesulfonic acid tetrabutylammonium salt in 3 ml of 30% acetone-water, the solution was added to Dowex 50W-X2. Passed through a (K ⁺ type) column and eluted with the same solvent. 95 mg of the product obtained by evaporating all the eluates in vacuo was lyophilized to obtain an amorphous powder having a melting point of 120 ° to 135 ° C.

Analysis of C ₁₀ H ₁₁ N ₂ O ₆ S ₂ K

Theoretic value: C 33.51; H 3.09; Found: C 33.46; H 3.08;

N 7.82; S 17.89 N 7.92; S 17.64

Example 75

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(carboxymethoxy) imino] acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[2-oxo-2- (phenylmethoxy) ethoxy] imino] acetyl] amino] -2-oxo-1 0.1 g of azetidinesulfonic acid, potassium salt (see Example 71) was dissolved in a mixture of 5 ml and 5 ml of ethanol and hydrogenated at room temperature in the presence of 0.2 g of 10% palladium based on activated carbon. After 2 hours, the catalyst was filtered off, and the remaining solution was lyophilized to obtain a target compound having a melting point of 235 占 폚 (decomposition).

Example 76

3-[[(S)-[(aminocarbonyl) amino] -2-thienylacetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid potassium salt, isomer A

71 μl (0.888 mmol) of pyridine in a solution of 277 mg of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 46, Method II, Part A) dissolved in 15 ml of anhydrous acetonitrile. ) And 166 mg of (D) 2-amino-4- (2-thienyl) -5- (4R) -oxazoline hydrochloride were added at -20 ° C. The mixture was stirred for 10 minutes, and 71 μl of pyridine (2nd aliquot) and 166 mg of oxazoline were added. After stirring for 10 minutes, the solvent was removed under reduced pressure, and the residue was dissolved in a water-acetone mixture, and then passed through an ion exchange resin (AG 50 W-X 2 20 ml, K ⁺ type, 200 to 400 mesh). Water was removed from the fractions 2 to 3 to obtain 248 mg of a mixture of single and right isomers.

The product was purified and the left and right isomers were separated by Dianion AG HP 20 (130 ml) and eluted with water. Isomer A was eluted at 30 mg in fractions 23-28 and 30 ml (8 mg fraction) in fractions 35-45. The intermediate fraction (10 mg) was combined with the other fractions to isolate 35 mg of isomer A and 59 mg of isomer B as a whole.

Analysis for Isomer A (melting point 158 ° to 165 ° C.)

Theoretic value: C 33.51; H 3.29; N; 13.18

Found: C 30.95; H 2.97; N; 12.98

Analysis for Isomer B (melting point 160 ° to 170 ° C. (decomposition))

Theoretic value: C 31.05; H 3.29 Found: C 31.17; H 3.09;

N 13.18; S 15.05 N 13.13; S 15.09

Example 77

3-methoxy-2-oxo-3-[(phenylsulfoacetyl) amino] -1-azetidinesulfonic acid, dipotassium salt

0.53 ml of anhydrous pyridine was added to a stirring solution of 366 mg of crude 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 74A) and 38 mg of borax dissolved in 35 ml of anhydrous acetonitrile. A solution of 348 mg of α-sulfophenylacetyl chloride monoetherate dissolved in 8 ml of acetonitrile was then added over 2 minutes. After 20 minutes, the solvent was removed in vacuo and the residue was treated with 35 ml of 0.5 M potassium phosphate buffer (pH 5.5). 383 mg of hydrogen tetrabutylammonium sulfate was added thereto, and the mixture was extracted three times with methylene chloride. The methylene chloride was dried over sodium sulfate and evaporated to give 685 mg of crude product.

This crude product was combined with 115 mg of the crude product obtained by the second run and the combined material was transferred to a SiliCAR CC-4 column using methylene chloride as eluent followed by 2, 4, 6, 8 and 10% methanol in methylene chloride. Purified. The product, a mixture of racemic single-sided isomers in the form of tetrabutylammonium (about 6: 1), was passed through a Dowex 50W-X2 (type K ⁺ ) resin using 20% acetone in water as solvent to convert to potassium salt. The product was lyophilized to obtain 171 mg of the target compound having a melting point of 205 ° to 210 ° C. (decomposition).

Analysis for _{C 12 H 12 N 2 O 9} S 2 K 2 · H 2 O

Theoretic value: C 29.51; H 2.89 Found: C 29.45; H 2.74;

N 5.74; S 13.10 N 5.51; S 12.82

Example 78

3-[(carboxyphenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, dipotassium salt

3-[[1,3-dioxo-2-phenyl-3- (phenylmethoxy) propyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid potassium salt (see Example 51) 39 mg Was dissolved in 5 ml of methanol. To this was added 3.9 mg of anhydrous potassium carbonate and 19 mg of 10% palladium based on activated carbon, and the mixture was hydrogenated at atmospheric pressure for 20 minutes. The catalyst was removed by filtration, and 34 mg of the clear residue obtained by evaporation of the filtrate in vacuo was lyophilized to give an amorphous powder having a melting point of 178 ° to 190 ° C. (decomposition).

Analysis for _{C 13 H 12 O 8 N 2} SK 2 · 0.5H 2 O

Theoretical: C 35.20; H 3.18 Found: C 35.51; H 2.96;

N 6.32; S 7.23 N 6.39; S 6.92

Example 79

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[2- (1,1-dimethylethoxy) -1- (methyltheo) -2-oxoethoxy] imino ] Acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

(Z) -2-amino-α-[[2- (1,1-dimethylethoxy) -1- (instead of (Z) -2-amino-α- (hydroxyimino) -4-thiazole acetic acid It carried out according to the method of Example 73 using methylthio) -2-oxoethoxy] imino] -4-thiazole acetic acid, and obtained the target compound of melting | fusing point 130 degreeC (decomposition).

Example 80

(±) -3-butoxy-3-[[(phenylmethoxy) carbonyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

185 mg of 2-oxo-3- [N-chloro-N-[(phenylmethoxy) carbonyl] amino] -1-azetidine sulfonic acid, tetrabutylammonium salt (see Example 49A) dissolved in 1 ml of dimethylformamide Was cooled to -78 ° C, and 3.8 ml of 0.73N lithium N-butoxide dissolved in n-butanol was added at -78 ° C. After 15 minutes, 0.5 M monobasic potassium phosphate buffer was added and the product was extracted three times with 40 ml of dichloromethane, then dried over sodium sulfate, filtered and concentrated in vacuo to give 179 mg of the corresponding tetrabutylammonium salt of the desired compound.

To a solution of 109 mg of tetrabutylammonium salt dissolved in acetone, 60 mg of perfluorobutyl sulfonic acid and potassium salt dissolved in acetone were added. The solvent was removed in vacuo and ethyl acetate was added thereto. The crystallized products were combined and dried to give 66 mg of the target compound having a melting point of 186.5 ° to 187.5 ° C (decomposition).

Example 81

[3 ± (E)]-3-methoxy-3-[[(methoxyamino) [2-[[(phenylmethoxy) carbonyl] amino] -4-thiazolyl] acetyl] amino-2-oxo -1-azetidinesulfonic acid, potassium salt

A suspension of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 46, Method II, Part A), 0.175 mmol and sodium borate 0.0175 mmol suspended in 2 ml of dichloromethane React with 28 μl of pyridine and 0.175 mmol of (E) -α- (methoxyimino) -2-[[(phenylmethoxy) carbonyl] amino] -4-thiazolylacetyl chloride at 0 ° C. and after 1 h this mixture Was diluted with dichloromethane and then quenched with water. The organic layer was washed with water, the saturated salts were dried and evaporated in vacuo. The residue was purified with 20 g of Mallinckrodt SiliCAR CC-4 silica gel to give the corresponding tetrabutylammonium salt of the desired compound.

43 mg of this tetrabutylammonium salt was dissolved in 0.5 ml of acetone, and 20 mg of potassium sulfate perfluorobutane dissolved in 0.5 ml of acetone was added thereto. 3 ml of ether was added thereto, and the resulting solid was collected and dried in vacuo to obtain 28 mg of the target compound having a melting point of 144 ° to 136 ° C. (decomposition).

Example 82

[3 ± (Z)]-3-methoxy-3-[[(methoxyimino) [2-[[(phenylmethoxy) carbonyl] amino] -4-thiazolyl] acetyl] amino] -2- Oxo-1-azetidinesulfonic acid, potassium salt

(E) -α- (methoxyimino) -2-[[(phenylmethoxy) carbonyl] amino] -4-thiazolylacetyl chloride instead of (Z) -α- (methoxyimino) -2- [ It carried out according to the method of Example 81 using [(phenylmethoxy) carbonyl] amino] -4-thiazolylacetyl chloride to obtain a target compound having a melting point of 168 ° to 136 ° C. (decomposition).

Example 83

3-[[(R) -α-[[(4-ethyl-2,3-dioxo-1-piperazinyl) -carbonyl] amino] phenylacetyl] amino] -3-methoxy-2-oxo -1-azetidinesulfonic acid, potassium salt

242 μl of anhydrous pyridine was added to a solution of 0.69 mmol of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 46, Method II, Part A) in 30 ml of anhydrous acetonitrile. A solution of 352 mg of (R) -α-[[(4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] amino] phenylacetyl chloride was added followed by dissolution in 4 ml of acetonitrile. After 1 hour, 84 μl of pyridine was added thereto, and 117 mg of the above-described acid chloride dissolved in 1 ml of acetonitrile was further added. The reaction was stirred for 20 minutes, diluted with 24 ml of 0.5M potassium phosphate monobasic buffer (pH5.5), and then concentrated in vacuo to remove acetonitrile. The aqueous residue was extracted three times with methylene chloride and the extracts were combined, dried over Na ₂ SO ₃ and evaporated to give 546 mg of residue. Two fractions (285 mg and 173 mg) containing the corresponding tetrabutylammonium salt of the desired compound by passing the residue through a silica gel column using methylene chloride as eluent followed by 2%, 4% and 6% methanol in methylene chloride. Got.

173 mg of the fraction was passed through 4.5 g of Dowex 50-X2 (K ⁺ ) resin using acetone-water as the eluent to obtain 119 mg of the target compound. 104 mg of this material was eluted sequentially with water, 5% acetone in water and 10% acetone in water as an eluent using HP20-AG resin column in water. Got it. 60 mg of the fractions were freeze-dried to obtain a solid having a melting point of 171 ° to 172 ° C. (decomposition).

Example 84

N- (3-butoxy-2-oxo-1-sulfo-3-azetidinyl) -2-phenyl-acetamide, tetrabutylammonium salt

2-oxo-3- [N-chloro-N [(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, instead of tetrabutylammonium salt, N-chloro-N- (2-oxo-1-sulfo- Using the 3-azetidinyl) -2-phenylacetamide and tetrabutylammonium salt (refer the corresponding potassium salt method in Example 47), it carried out according to the method of Example 80 and obtained the oily target compound.

NMR (CDCl ₃ )

3.62 (s, 2H, C ₆ H ₅ CH ₂ )

4.03 (ABq, 2H, ν = 7cps, C-4CH ₂ )

6.98 (s, 1 H, NH)

7.30 ppm (s, 5H, C ₆ H ₅ )

Example 85

(R) -3-methoxy-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

Trimethylsilyl chlorosulfonate was slowly added to dimethylformamide at 0 ° C. and then discharged at 0.1 mmHg for 30 minutes at 0 to 25 ° C. to prepare a 1 M solution of dimethylformamide-sulfur trioxide complex. Under argon atmosphere, 50 mg of (R) -N- (3-methoxy-2-oxo-1-azetidinylphenyl) -acetamide was dissolved in 0.2 ml of anhydrous dimethylformamide, and cooled to 0 ° C. To this was added 0.428 ml of cooled 1M dimethylformamide-antigen trioxide solution, and the mixture was stirred for 2 hours and poured into 15 ml of 0.5N monobasic potassium phosphate. The solution was extracted twice with dichloromethane and 73 mg of tetrabutylammonium disulfide was added thereto. Extracted three times with 10 ml each of dichloromethane, dried and evaporated in vacuo to give a viscous oily substance. Chromatography with Mallincrodt CC-4 silica gel (50: 1) using 2% methanol in dichloromethane as eluent gave (R) -3-methoxy-2-oxo-3-[(phenylacetyl) amino ] -1-azetidinesulfonic acid and 34 mg of tetrabutylammonium salts were obtained. Ion exchange reaction was carried out with Dowex 50-X2 (K ⁺ , 10 equivalents), and the aqueous eluate was lyophilized to obtain potassium salt of the target compound. Melting point 130 ° C. (decomposition), [α] _D = + 52 ° (C = 0.5, water)

Example 86

(S) -3-[[[[(1-ethyl-4-hydroxy-3-methyl-pyrazolo- [3,4-b] pyridin-5-yl) carbonyl] amino] phenylacetyl] amino] 2-oxo-1-azetidinesulfonic acid, potassium salt

Α-[[(1-ethyl-4-hydroxy-3-methyl-1H-pyrazolo- [3,4 instead of (Z) -2-amino-α- (hydroxyimino) -4-thiazole acetic acid -b] pyridin-5-yl) carbonyl] amino] benzeneacetic acid was carried out according to the method of Example 73 to obtain the target compound at melting point of 233 ° -236 ° C (decomposition).

Example 87

(R) -3-acetylamino-3-methoxy-2-oxo-1-azetidinesulfonic acid potassium salt

A) 3-acetylamino-1- [1-carboxy-2-methyl (propyl)]-(3R) -3-methoxy-2-oxoazetidine

(6R-cis) -7-acetylamino-7-methoxy-3-methyl-8-oxo-5-thia-1-azabicyclo [4,2,0] oct-2-ene-2 dissolved in water To a solution of 650 mg of carboxylic acid and 191 mg of sodium bicarbonate was added 11 ml of a slurry of conventional Raney nickel (0.6 g / ml) washed with water until neutral. The mixture was immersed in an oil bath preheated to 170 ° C. and refluxed for 2-3 minutes (at which time the temperature of the bath was maintained between 150 ° and 170 ° C.). After refluxing for 15 minutes, the reaction mass was quenched by cooling in an ice bath. The catalyst was removed by filtration through Celite and the pH was adjusted to 2 by adding 1N hydrochloric acid to the filtrate (pH11). The aqueous solution was extracted five times with ethyl acetate, the extracts were combined, dried over Na ₂ SO ₄ , and the solvent was removed in vacuo to give 487 mg of an oily substance. Chromatography with silica gel eluted with chloroform to give 381 mg of an oily product.

B) 3-acetylamino-1- [1- (acetyloxy) -2-methyl (propyl)]-(3R) -3-methoxy-2-oxoazetidine

464 mg of azetidinone was dissolved in 15 ml of anhydrous acetonitrile, and this solution was washed with argon for 15 minutes, and then 359 mg of copper acetate was added thereto, stirred for 1 minute to dissolve the salt, and 797 mg of lead tetraacetate Was added. The flask was immersed in an oil bath preheated and maintained at 55 ° to 65 ° C. for about 15 minutes while continuously injecting argon into the mixture to raise the reaction temperature. The mixture was cooled to room temperature, the celite was filtered off and the filtration pad was washed well with acetonitrile. The solvent was removed in vacuo from the combined filtrate and the wash. The residue was dissolved in water and extracted four times with ethyl acetate. The extracts were combined, dried over Na ₂ SO ₄ , and the solvent was removed in vacuo to give 382 mg of an oily target compound.

C) (R) -N- (3-methoxy-2-oxo-1-azetidinyl) acetamide

The oily substance was dissolved in a methanol (10 ml): water (1 ml) mixture and cooled in an ice-methanol bath at -10 ° to -15 ° C, then 194 mg of potassium carbonate was added thereto, followed by 53 mg of sodium borohydride. Added. It was stirred for 110 min at -15 [deg.] C. to -8 [deg.] C., then the solvent was removed in vacuo, the residue was dissolved in water and then adjusted to pH6 by addition of 1N hydrochloric acid. The mixture was extracted with ethyl acetate, dried over Na ₂ SO ₄ , and the solvent was removed in vacuo to yield 224 mg of an oily substance. This oily substance was chromatographed with silica gel, and eluted with 5% methanol: 95% methylene chloride to obtain 169 mg of the oily substance. This target compound was crystallized with ether-pentane to give 131 mg of a material having a melting point of 106 ° to 112 ° C. (sintered at 103.5 ° C.).

D) (R) -3-acetylamino-3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

Under argon atmosphere, 50 mg of (R) -3-acetylamino-3-methoxy-2-oxo-1-azetidine was placed in a flask and cooled to 0 ° C. To this was added 1M solution of dimethyl formamide-antioxidant complex dissolved in 0.95 ml of dimethylformamide, followed by stirring for 15 minutes. The contents in the flask were then poured into 40 ml of 0.5NK ₂ HPO ₄ solution and extracted twice with 10 ml of methylene chloride. Tetrabutylammonium sulfate (1.2 equivalents) was added to this aqueous solution, and the resulting mixture was extracted four times with 10 ml of methylene chloride. This extract was dried over Na ₂ S0 ₄ and concentrated to give 39 mg of product. The tetrabutylammonium salt was passed through a Dowex 50-X ² (K ⁺ ) column to convert to the potassium salt of the desired compound. The aqueous fraction was concentrated to give 19 mg of potassium salt having the same NMR spectrum as that prepared in Example 46Ib and isolated from natural source (Example 165).

Example 88

(±) -3-[(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

A) (±) -3-[(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

202 mg of 3-amino-3-methoxy-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 7A) were dissolved in 20 ml of anhydrous acetonitrile and stirred well at -20 ° C under a dry nitrogen atmosphere. To 167 μl of anhydrous pyridine and 96 μl of α-azidophenylacetyl chloride were added. After 20 minutes, 12 ml of 0.5 M monobasic potassium phosphate buffer (pH 5.5) was added and acetonitrile was removed in vacuo. The aqueous residue was extracted three times with methylene chloride, and the extract was dried over anhydrous sodium sulfate and then evaporated in vacuo to give 281 mg of crude rubbery product. This was chromatographed on a silica gel (30 g) column using a mixture of methylene chloride and methylene chloride-methanol to 6% methanol as eluent to obtain 231 mg of the target compound.

B) (±) -3-[(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

30% acetone: 231 mg of (±) -3-[(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt dissolved in 15 ml of water was added to Dowex 50W-X2 ( K ⁺ type, 3 ml) was passed through a column and eluted with water. Eluate AllCell was evaporated in vacuo to give 168 mg of a 1: 1 mixture of colorless glassy single-sided isomers. This material was passed through a 60 ml HP20-AG column and eluted with water and water: 10% acetone to give 71 mg of a 1: 1 mixture of racemic single-sided isomers and 70 mg of a 1: 1 mixture of racemic single-sided isomers. The 1: 1 mixture was lyophilized and evaporated in vacuo at 40 ° C. to give a dried product, in the form of hemihydrate, of melting point 130 ° C. (decomposition).

Analysis for _{C 12 H 12 N 5 O 6} · K · 0.5H 2 O

Theoretic value: C 35.90; H 3.26; Found: C 35.94; H 3.07;

N 17.45; S 7.98 N 17.24; S 8.02

Example 89

3-[(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt isomer A

Isomer A was crystallized by leaving a 1: 3 mixture of racemic single-sided isomers obtained in Example 88B in deuterated at room temperature. After freezing this, the mother liquor was removed and 28 mg of the crystals were dried in vacuo at 40 ° C. to give the desired compound in the form of mono-deuterium at melting point 130 ° C. (decomposition).

Example 91

[3S (Z)]-3 [[(methoxyimino) [2-[[(phenylmethoxy) carbonyl] amino] -4-thiazolyl] acetyl] amino] -2-oxo-1-azetidinesulfonic acid Potassium salt

In a solution of (S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) and 0.170 mmol of sodium borate dissolved in 2 ml of methylene chloride, 62 μl of pyridine at 0 ° C. and ( 0.51 mmol of Z) -α- (methoxyimino) -2-[[(phenylmethoxy) carbonyl] amino] -4-thiazoleacetyl chloride was added. After 40 minutes the reaction mixture was diluted with methylene chloride and water and then diluted with 5.1 ml of 0.1 M tetrabutylammonium sulfate buffered to pH4. The organic layer was separated, washed with water adjusted to pH 2, water adjusted to pH 7 and water saturated with sodium chloride, dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified with 10 g of Silic AR CC-4 silica gel and the product eluted with 10% methanol / methylene chloride.

The tetrabutylammonium salt was dissolved in acetone / water and then passed through 8 ml of ion exchange resin (AG 50 W-X 2, K ⁺ type, 100-200 mesh). Water was removed in vacuo from fractions 1 to 2 to obtain 40 mg of the target compound having a melting point of 172 ° to 174 ° C. (decomposition).

Analysis of C ₁₇ H ₁₆ N ₅ O ₈ S ₂ KH ₂ O

Theoretic value: C 37.84; H 3.33; Found: C 37.95; H 3.30;

N 12.99; S 11.87 N 12.73; S 11.53

Example 92

(±) -3-butoxy-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

A) 3- [chloro (phenylacetyl) amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

5.25% of a solution of 350 mg of (S) -2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidine sulfonic acid and tetrabutylammonium salt (see Example 4) dissolved in 3 ml of methylene chloride To a suspension of 1.27 g of sodium borate suspended in 4.72 ml of sodium hypochlorite solution and 20 ml of water was added at 0 ° C. After 1 hour, 25 ml of 0.5 M monobasic potassium phosphate was added thereto, and the mixture was extracted three times with 50 ml of methylene chloride. The organic extract was dried over sodium sulfate, filtered and concentrated in vacuo to give 344 mg of the desired compound.

B) (±) -3-butoxy-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, tetrabutylammonium salt

0.73 N n dissolved in 6 ml of n-butanol in 1 ml of dimethylformamide, a solution of 344 g of 3-[[chloro (phenylmethoxy) carbonyl] amino-1-azetidinesulfonic acid and tetrabutylammonium salt dissolved in 5 ml of dimethylformamide was dissolved. The lithium butoxide solution was added at -78 ° C under inert atmosphere. After 10 minutes, the mixture was diluted with 175 ml of 0.5 M monobasic potassium phosphate solution, extracted three times with methylene chloride, and then the organic extract was dried over sodium sulfate, filtered and the solvent was removed in vacuo. The residue was purified with 80 g of Silic AR CC-4 silica gel and then 130 mg of the target compound was eluted with 4-8% methanol in methylene chloride.

C) (±) -3-butoxy-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

43 mg of (±) -3-butoxy-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid and tetrabutylammonium salt were dissolved in a water-acetone mixture (9: 1), and cation exchange column (Dowex AGMP 50W-X2 100-200 mesh, 5 g, K ⁺ type). The product was eluted with water, and the eluate was concentrated in vacuo to give 20 mg of the target compound having a melting point of 122 ° to 125 ° C.

Theoretic value: C 44.66; H 4.96; Found: C 44.77; H 4.76;

N 6.95; S 7.94 N 6.76; S 7.75

Example 93

(±) -3-ethoxy-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

200 mg of 3- [chloro (phenylacetyl) amino] -2-oxo-1-azetidinesulfonic acid and potassium salt (see Example 92A) dissolved in 4 ml of dimethylformamide are dissolved in 12.20 ml of ethanol under an inert atmosphere at -78 ° C. To 0.5 N lithium ethoxide. After 10 minutes, the mixture was diluted with 15 ml of 0.5 M monobasic potassium phosphate solution. After extraction three times with methylene chloride, the organic layer was dried over sodium sulfate, filtered and the solvent was removed in vacuo. The residue was purified with 20 g of Silic ARCC-420 g silica gel and 40 mg of tetrabutylammonium salt of the product was eluted with 2% methanol in methylene chloride.

The tetrabutylammonium salt was dissolved in a water-acetone mixture (9: 1), passed through a cation exchange column (Dowex AGMP 50W-X2 100-200 mesh, 5 g, K ⁺ type) to elute the product with water, and the eluate was dried in vacuo. It concentrated and 25 mg of target compounds of melting | fusing point 94 degrees-96 degrees were obtained.

Analysis of C ₁₃ H ₁₅ N ₂ O ₆ SK

Theoretic value: C 42.62; H 4.10; Found: C 40.36; H 3.66;

N 7.65; S 8.74 N 6.77 S 8.44

Example 95

[3R (R ^* ) and 3S (S ^* )]-3-[[[(aminocarbonyl) amino] phenylacetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

A) (±) -3-[(aminophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, intramolecular salt

(±) -3-[(azidophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid and potassium salt (see Example 88) were dissolved in 60 ml of anhydrous methanol, and 0.6 ml of trifluoroacetic anhydride and 105 mg of 10% palladium based on activated carbon were added followed by hydrogenation of the mixture for 1 hour. The catalyst was filtered off and the filtrate was evaporated in vacuo to give 271 mg of crude product.

B) [3R (R ^* ) and 3S (S _* )]-3-[[[(aminocarbonyl) amino] phenylacetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

(±) -3-[(aminophenylacetyl) amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid and 271 mg of potassium salt were dissolved in 6.5 ml of water, and 87 mg of potassium cyanate was added thereto. This mixture was then stirred at room temperature for 3 hours. The solution was concentrated to about 2 ml in vacuo and chromatographed with 100 ml HP20-AG column using water as eluent. After freeze drying, 29 mg of isomer A with a melting point of 160 ° C. (decomposition) was isolated.

Analysis of C ₃ H ₁₅ N ₄ O ₇ SK Monohydrate

Theoretic value: C 36.44; H 3.09; Found: C 36.35; H 3.79

N 13.07; S 7.48 N 12.81; S 7.31

Example 96

[3R (S ^* ) and 3S (R ^* )]-3-[[[(aminocarbonyl) amino] phenylacetyl] amino] -3-methoxy-2-oxo-1-azetidinesulfonic acid, potassium salt

[3R (R ^* ) and 3S (S ^* )]-3-[[[(aminocarbonyl) amino] phenylacetyl] amino] -3-methoxy-2-oxo-1-ase prepared in Example 95 [3R (S *) and 3S (R ^* )]-3-[[[(aminocarbonyl) amino] phenylacetyl] amino] -3-methoxy-2-oxo-1- with thidinesulfonic acid and potassium salt Azetidinesulfonic acid and potassium salt were prepared by lyophilization. Melting point 160 ° C. (decomposition).

Analysis of C ₃ H ₁₅ N ₄ O ₆ SK Sesquihydrate

Theoretic value: C 35.69; H 4.14; Found: C 35.81; H 3.98

N 12.81; S 7.33 N 12.50; S 7.32

Example 97

[3 ± (S ^* )]-3-methoxy-3-[[[[[2-oxo-3- (phenylmethylene) amino] -1-imidazolidinyl] carbonyl] amino] -2-thier Nylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

306 mg of 3-amino-3-methoxy-2-oxo-azetidinesulfonic acid, tetrabutylammonium salt (crude containing 274 mg of organic material, prepared as described in Example 74) dissolved in 20 ml of anhydrous acetonitrile 0.30 ml of anhydrous pyridine (3.72 mmol) was added to the stirred solution, and then partially dissolved and suspended in 10 ml of anhydrous acetonitrile (S)-[[[2-oxo-3-phenylmethylene) amino] -1-imida 484 mg of zolidinyl] carbonyl] amino] -2-thienylacetylchloride were added. The reaction was stirred and then the temperature was raised to 0 ° C. over 1 hour.

The reaction was diluted with a large amount of methylene chloride and then 22 ml of 0.5 M potassium phosphate monobasic buffer (Ph 5.5). The aqueous layer was washed with methylene chloride, the combined methylene chloride extracts were washed with water, dried over Na ₂ SO ₄ and evaporated to give 508 mg of residue. This residue was chromatographed with Silic ARCC-450g using methylene chloride and 2% and 4% methanol in methylene chloride as eluent to give 251 mg of tetrabutylammonium salt of the target compound.

To 251 mg of this salt dissolved in acetone was added a solution of 107 mg of potassium fluorobutanesulfonic acid salt dissolved in a few ml of acetone. The precipitate formed by adding ethyl acetate was washed three times with ethyl acetate by centrifugation, which was dried for 2 hours at 40 ° C / 1 mm in vacuo to give 95 mg of the desired potassium salt at a melting point of 200 ° C (decomposition). About 1: 2 mixture of isomers obtained the form.

Analysis of C ₂₁ H ₂₁ N ₆ O ₈ S ₂ K

Theoretic value: C 42.85; H 3.60; Found: C 43.92; H 3.74;

N 14.28; S 10.87 N 13.94; S 10.71

Example 98

(± -cis) -4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

A) N-benzyloxy-t-boc ^* -allothreonineamide

A solution of free amine was added from 6.9 g of 1, t-boc-allothroonine and 5.3 g of O-benzylhydroxylamine HCl (about 0.033 mol, ethyl acetate-sodium bicarbonate glass), which was dissolved in 80 ml of tetrafuran and tetra- 4.82 g of hydroxybenzotriazole and 6.5 g of dicyclohexylcarbodiimide in 20 ml of tetrahydrofuran. After stirring at room temperature for about 16 hours, the slurry was filtered, concentrated in vacuo and then with a 400 ml silica gel column. I did kramotography. Elution with 5-10% ethyl acetate / chloroform gave 6.8 g of the target compound in fractions 7-22 (200 ml each).

^* "boc" is butoxycarbonyl.

B) (± -cis) -N-benzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone

A solution of 6.8 g of N-benzyloxy-t-boc-allothroonylamide dissolved in 200 ml of tetrahydrofuran was added by adding 5.24 g of triphenylphosphine and 3.2 ml of diethylazodicarboxylate, and stirring for about 16 hours. I broke it. The solvent was evaporated in vacuo and the residue was chromatographed on a 500 ml silica gel column. Elution with methylene chloride was followed by recrystallization with ether to give a total of 2.65 g of azetidinone. The mother liquor and mixed fractions were rechromatography to yield 0.6 g more. Crystallization twice with ether (-20 ° C) gave a sample for analysis of the target compound at a melting point of 140 ° to 142 ° C.

C) (± -cis) -3-t-butoxycarbonylamino-1-hydroxy-4-methylazetidinone

To a solution of 3.2 g of cis-N-benzyloxy-3-t-butoxy-carbonylamino-4-methylazetidinone dissolved in 200 ml of 95% ethane, 0.7 g of activated carbon based 10% palladium was added under a hydrogen atmosphere. And stirred. After 40 minutes, the slurry was filtered (249 ml of filtration), the filtrate was evaporated, and then titrated with ether to give 2.05 g of solid product having a melting point of 134 ° to 136 ° C. as two outputs.

D) (± -TLTM) -3-t-butoxycarbonylamino-4-methylazetidinone

In a solution of 2.05 g of cis-3-t-butyloxycarbonylamino-1-hydroxy-4-methylazetidinone dissolved in 60 ml of methane, a total of 90 ml of 4.5 M ammonium acetate (40, 20 and 30 ml each) and titanium Trichloride (20, 10 and 15 ml each) was treated with a total of 45 ml and the second and third additions were done after 15 and 120 minutes respectively. After 135 minutes, the solution was diluted with the same volume of 8% sodium chloride and extracted three times with 300 ml of ethyl acetate. The combined organic layers were washed with a mixture of 100 ml each of 5% sodium bicarbonate and saturated salt, dried and evaporated. This was titrated with ether to give 1.65 g of solid product as two outputs. The first product was recrystallized with ether to obtain an analytical sample having a melting point of 176 to 178.5 ° C.

E) (± -cis) -3-benzyloxycarbonylamino-4-methylazetidinone

A solution of 1.55 g of cis-3-t-butoxycarbonylamino-4-methylazetidinone dissolved in 4 ml of methylene chloride and anisol, respectively, was cooled to 0 ° C., and 50 ml of trifluoroacetic acid was added thereto. . After 90 minutes, the solvent was evaporated in vacuo (evaporated three times with the addition of benzene). The residue was dissolved in 25 ml of acetone, and 5% sodium bicarbonate was added to raise the initial pH (2.5) to 7, followed by 2 ml of benzylchloroformate. The solution was kept at 0 ° C. and pH 7 for 4 hours and then the slurry obtained by removing acetone in vacuo was filtered. The filtrate was saturated with salt and extracted with methylene chloride. The obtained solid was dissolved in methylene chloride, dried, the organic layers were combined and concentrated, and the residue was chromatographed with a 200 ml silica gel column. Elution with chloroform: ethyl acetate (3: 1) gave 850 mg of the target compound in fractions 4-11 (100 ml each). A small amount of sample was crystallized with ether and used as analytical sample. Melting point 165 ° to 166 ° C.

F) (± -cis) -4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

Cis-3-benzyloxycarbonyl amino-4- suspended in 7 ml each of dimethylformamide (dehydrated with 4A form and activated for 15 hours at 320 ° C. under argon) and methylene chloride (dried with Al ₂ O ₃ ). To a suspension of 0.75 g of methylazetidinone, 1.66 g of pyridine-sulfur trioxide complex was added. After stirring for 3 hours at room temperature in the presence of nitrogen, 1.66 g of pyridine-sulfur trioxide complex was further added. Subsequently, the reaction mixture was stirred at room temperature in the presence of nitrogen for about 16 hours, and then 4.6 g of the residue obtained by removing dimethylformamide in vacuo was dissolved in 300 ml of 0.5 M monobasic potassium phosphate solution at 40 ° C. for 10-15 minutes. . The solution was cooled and passed through a HP20 resin (3 cm x 60 cm) column using 400 ml dibasic water (400 ml) of 0.5 m monobasic potassium phosphate as eluent and water: acetone (14: 1) to give the product in fractions 13-26 (100 ml each). 280 mg was obtained. Crystallization with methanol: petroleum ether gave 757.5 mg of analytical sample. Melting point 214 ° to 215.5 ° C. (decomposition).

Analysis of C ₁₂ H ₁₃ N ₂ SO ₆ K

Theoretic value: C 40.90; H 3.72; Found: C 40.43; H 3.60;

N 7.95; S 9.10; N 7.89; S 8.69

K 11.10 K 10.82

Example 99

(3S-trans-4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

It carried out according to the method of Example 98 using 1-t-boc-threonine instead of d, l-t-boc-allothroonyl, and the target compound of melting | fusing point 133 degreeC-135 degreeC was obtained.

Analysis of C ₁₂ H ₁₃ N ₂ O ₆ SK

Theoretic value: C 40.90; H 3.72; Found: C 40.72; H 3.60

N 7.95; S 9.10; N 7.99; S 8.80;

K 11.10 K 10.82

Example 100

(3S-trans) -4-methyl-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

A) (3S) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

Dissolve 352.4 mg of (3S-trans) -4-methyl-2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and potassium salt (see Example 99) in 20 ml of distilled water. It was treated with 373.5 mg (1 mmol) of hydrogen tetrabutylammonium sulfate. After stirring for 10 minutes at room temperature, the solution was saturated with sodium chloride and then extracted three times with 10 ml each of methylene chloride. Methylene chloride was dried over sodium sulfate and 536 mg of tetrabutyl ammonium salt obtained by evaporation in vacuo was hydrogenated with 270 mg of 10% palladium based on activated carbon in 25 ml of dimethylformamide. The mixture was filtered through celite and washed twice with 2.5 ml of dimethylformamide to obtain the target compound in solution form.

B) (3S-trans) -4-methyl-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

206 mg of crude (3S) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (combined with the filtrate and the wash solution) prepared at the above part A at 0 ° C. , 153 mg of N-hydroxybenzotriazole and 138 mg of phenylacetic acid and the reaction mixture was stirred at 0 ° C. for 1 hour and then at room temperature for 2 hours. The resulting precipitate was filtered off, the filtrate was evaporated in vacuo and the residue was dissolved in 10 ml of acetone and filtered. The filtrate was treated with 25 ml of acetone saturated with potassium iodide, followed by 200 ml of ether. 752.7 mg of product solid was a mixture of potassium salt and tetra butylammonium salt of the desired compound. This solid was dissolved in 50 ml of 0.5 M monobasic potassium phosphate solution, and several fractions obtained by eluting with water followed by water-acetone using an HP-20 column were combined and evaporated to give a purified tetrabutylammonium salt. An aqueous solution of this material was passed through Dowex 50W-X2 (type K ⁺ ) to give 121.4 mg of potassium salt of the target compound. This was titrated with acetone-hexane to obtain 104.6 mg of the target compound having a melting point of 211 ° to 213 ° C.

Analysis of C ₁₂ H ₁₃ N ₂ O ₅ SK · ½H ₂ O

Theoretic value: C 41.72; H 4.09; Found: C 41.70; H 4.01;

N 8.11; S 9.28; N 8.07; S 9.01;

K 11.32 K 11.02

Example 101

(Cis) -4-methyl-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

(± -cis) -4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt (see Example 98) 320 mg of hydrogen tetrabutylammonium sulfate It was dissolved in 20 ml of water containing 483 mg, and then adjusted to pH5.5. The solution was extracted six times with 25 ml of methylene chloride to give 517.3 mg of an oily substance. To the solution dissolved in 15 ml of dimethylformamide, 400 mg of 10% palladium based on activated carbon was added under a hydrogen atmosphere and stirred for 90 minutes. The catalyst was filtered off, and 150 mg of phenyl acetate, 169 mg of N-hydroxybenzotriazole and 247 mg of dicyclohexylcarbodiimide were added to the filtrate and stirred for 7.5 hours. The solvent was removed in vacuo and the residue was dissolved in 20 ml of acetone and then filtered. The filtrate was treated with 25 ml of a 0.044 M potassium iodide solution in acetone. After dilution with an equal volume of ether, the resulting solids (330 mg) were eluted with 200 ml of water followed by acetone-water (1: 9) using a 50 ml HP-20 column in 20 ml of 0.05 M monobasic potassium phosphate solution. Ridon positive material was obtained from fractions 6-10 (50 ml). Fractions 7-9 were evaporated to yield 81 mg of solid. 46 mg of the target compound was obtained, which was recrystallized from acetonitrile-water and decomposed at 205 캜 or higher. Another 6 mg of output 2 was obtained in the filtrate, and fractions 6 and 10 were evaporated and recrystallized to give an additional 5 mg.

Analysis of C ₁₂ H ₁₃ N ₂ O ₅ SK

Theoretic value: C 42.84; H 3.89; Found: C 42.75; H 3.82;

N 8.33; S 9.53; N 8.32; S 9.26

K 11.62 K 11.63

Example 102

[3S- [3α (Z), 4β] -3-[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-methyl-2-oxo-L-azetidinesulfonic acid, potassium salt

A) (3S-trans) -4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt

Dissolve 352.4 mg of (3S-trans) -4-methyl-2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and potassium salt (see Example 99) in 20 ml of water To this was added 373.5 mg of hydrogen tetrabutylammonium sulfate. This aqueous solution was extracted three times with methylene chloride, and the extracts were combined and dried over sodium sulfate. The solvent was removed to obtain 534.6 mg of the target compound.

B) [3S- (3α (Z), 4β] -3-[[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-methyl-2-oxo-1-azetidine Sulfonic acid, potassium salt

A solution obtained by dissolving 534.6 mg of (3S-trans) -4-methyl-2-oxo-3-[[((phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and tetrabutyl ammonium salt in 20 ml of dimethylformamide was prepared. 26.3 ml of hydrogen was absorbed by hydrogenation with 220 mg of activated carbon based 10% palladium for 2.75 hours at atmospheric pressure. The mixture was filtered and washed twice with 2.5 ml of dimethylformamide. The filtrate and washings (about 25 ml total) were 161 mg of (Z) -α- (methoxyimino) -2-amino-4-thiazole acetic acid, 136 mg of N-hydroxybenzotriazole and dicyclohexylcarbodiimide in the presence of nitrogen Stir with 164.8 mg. The mixture was stirred for about 16 hours in the presence of nitrogen. In vacuo, dimethyl formamide was removed and the rubbery residue was dissolved in acetone and then filtered to remove urea. A solution in which 272 mg (0.8 mmol) of fifluorobutanesulfonic acid and potassium salt was dissolved in 0.8 ml of acetone was added to the filtrate. The slurry was diluted with the same volume of ether, filtered and 325.5 mg of the crude product obtained was purified by chromatography with 75 ml of HP-20AG. Eluent was eluted with 400 ml of water and 400 ml of water: acetone (9: 1) mixture (50 ml fractions) to give 335 mg of the product in fractions 3-10. Titration with acetone-hexane gave analytical samples in fractions 3-5. Fractions 6-10 were titrated in a similar manner to give 90.4 mg of solid product.

Analysis of C ₁₀ H ₁₂ N ₅ O ₆ S ₂ K

Theoretic value: C 29.92; H 3.01; Found: C 30.32; H 3.49;

N 17.45; S 15.97 N 15.82; S 13.95;

K 9.74 K 15.45

NMR (D ₂ O)

1.57 (3H, d, J = 7) 4.70 (1H, d, J =)

3.97 (3H, s) 6.95 (1H, s)

4.30 (1H, double q, J = 7.3)

Example 103

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl 2-oxo-1-azetidinesulfonic acid, dipotassium salt

A) N-benzyloxy-t-boc-threonineamide

N-hydroxybenzo dissolving a solution of this free amine in 100 ml of tetrahydrofuran from 8.76 g of t-boc-threonine and 6.4 g of O-benzylhydroxylamine HCl (ethyl acetate-sodium bicarbonate glass) 6.12 g of triazole and 8.24 g of dicyclohexylcarbodiimide. The mixture was stirred for 26 h in the presence of nitrogen, filtered and then evaporated in vacuo. The residue was chromatographed on 300 g of silica gel column, and eluted with chloroform and chloroform-ethyl acetate (3: 1) to obtain a compound 7.2 g. This compound was crystallized with ether-hexane to give 4.18 g of the target compound.

B) (3S-trans) -N-benzyloxy-3-t-buroxy carbonylamino-4-methylazetidinone

12.67 g of N-benzyloxy-t-boc-threonine amide, 11.5 g of triphenylphosphine and 6.23 ml of diethylazodicarboxylate were stirred for about 16 hours in the presence of nitrogen dissolved in 380 ml of tetrahydrofuran. This solution was evaporated and chromatographed on a 900 g silica gel column. 13.69 g of the compound obtained by eluting with chloroform-ethyl acetate (3: 1) was crystallized with ether-hexane to obtain 9.18 g of the target compound.

C) (3s-trans) -3-t-butoxycarbonylamino-1-hydroxy-4-methylazetidinone

A solution of 9.18 g of (3S-trans) -N-benzyloxy-3-t-butoxycarbonylamino-4-methylazetidinone in 300 ml of 95% ethanol was dissolved in 1.85 g of 10% palladium based on activated carbon under hydrogen atmosphere. Stir together. After 141 minutes, the slurry was filtered and evaporated in vacuo. The residue was recrystallized from ether-hexane to give 5.12 g of the target compound.

D) (3S-trans) -3-tbutoxycarbonylamino-4-methyl-azetidinone

A solution of 4.98 g of (3S-trans) -3-t-butoxycarbonylamino-1-hydroxy-4-methylazetidinone dissolved in 200 ml of methanone was treated with 132 ml of 4.5 M ammonium acetate followed by 1.5 M titanium Treatment with 66 ml of trichloride was followed by stirring for 4.5 hours. The aqueous solution was diluted with the same volume of 8% sodium chloride and extracted with ethyl acetate to give 3.48 g of crude product. This product was recrystallized from ether-hexane to give 3.3 g of the target compound.

E) (3S-trans) -3-benzyloxycarbonylamino-4-methylazetidinone

A solution of 3.3 g of (3S-trans) -3-t-butoxycarbonylamino-4-methyl azetidine dissolved in 10 ml of dichloromethane and anisol, respectively, was cooled to 0 ° C., followed by 112 ml of trifluoroacetic acid. Was added. The solution was stirred for 90 minutes and evaporated in vacuo (evaporated three times with the addition of benzene). The residue was dissolved in 70 ml of acetone and 5% sodium bicarbonate solution was added to this solution to adjust the pH of the solution to 7. 5.33 g of benzyl chloroformate at pH 6.5-7.5 were added over 1 hour. The mixture was stirred at pH 7 for 30 minutes, diluted with 100 ml of saturated salt and extracted with ethyl acetate (3 times 400 ml). The residue obtained by evaporation of the extract was chromatographed on a silica gel column for 1 L, and eluted with chloroform-ethyl acetate (4: 1) to obtain 2.19 g of a compound. Crystallization with ether-hexane afforded 1.125 g of the target compound.

F) (3S-trans) -4-methyl-2-oxo-3-[[(phenylmethoxy) -carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt

A solution of 600 mg of (3S-trans) -3-benzyloxycarbonylamino-4-methyl-azetidinone dissolved in 2 ml of dimethylformamide was cooled to 0 ° C., followed by 4 ml of 0.8 M sulfur trioxide solution in dimethylformamide. Was added. The solution was stirred for 1 hour at room temperature in the presence of nitrogen and poured into 80 ml of cooled 0.5 M monobasic potassium phosphate (adjusted to pH 5.5). The solution was extracted three times with 50 ml each of methylene chloride, and 868 mg of tetrabutylammonium disulfide was added thereto. The resulting solution was extracted four times with 75 ml of methylene chloride, and the organic layers were combined, washed with 8% aqueous sodium chloride solution, dried and evaporated in vacuo to give 1.54 g of the target compound.

G) [3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl)-[(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] Acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

A solution of 1.54 g of (3S-trans-4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid and tetrabutylammonium salt was dissolved in 45 ml of dimethylformamide. Under the atmosphere, the mixture was stirred for 2 hours with 800 mg of activated carbon based 10% palladium The catalyst was filtered off and the filtrate was filtered into (Z) -2-amino-α-[(1-diphenylmethoxycarbonyl-1-methylethoxy). 1.24 g amino] -4-thiazole acetic acid, 0.4 g N-hydroxybenzotriazole and 580 mg dicyclohexylcarbodiimide were added and stirred for about 16 hours The residue obtained by evaporation of the slurry in vacuo was acetone. Titrated to 20 ml, followed by filtration The combined filtrate and 2 ml of wash were treated with 868 mg of potassium perfluorobutanesulfonate dissolved in 3 ml of acetone, which was diluted with 75 ml of ether to give a solid, which was isolated by decanting the mother liquor. Then titrated with ether, filtered To obtain 0.91 g of the target compound, the mother liquor was further diluted with 100 ml of ether to obtain 0.45 g of the target compound as a second output.

H) [3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) amino) acetyl] amino] -4- Methyl-2-oxo-azetidinesulfonic acid, dipotassium salt

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] acetyl] amino ] A slurry of 140 mg of 4-methyl-2-oxo-1-azetidinesulfonic acid and potassium salt (first product) in 0.5 ml of anisole was stirred at -12 ° C. under a nitrogen atmosphere and then cooled therein ( -10 ° C.) trifluoroacetic acid was stirred and then warmed to room temperature. The solids were isolated by centrifugation and washed twice with ether. The pH of the solution dissolved in 5 ml of cold water was immediately adjusted to pH 5.5 by adding 0.4 N potassium hydroxide, eluted with water using an 80 ml HP-20AG column, and evaporated (addition of acetonitrile 3 Times), and then titrated with ether to give 72 mg of the target compound in fractions 7-11 (10 ml). Melting Point About 250 ° C (Decomposition)

Analysis of C ₁₃ H ₁₅ N ₅ O ₈ S ₂ K ₂

Theoretic value: C 30.51; H 2.95; Found: C 29.63; H 3.20;

N 13.69; S 12.53; N 12.96; S 11.94;

K 15.28 K 12.78

NMR (D ₂ O)

1.46 (s, 6H) 4.67 (1H, d, J = 2)

1.58 (1H, d, J = 7) 6.95 (s, 1H)

4.28 (1H, double doublet of doublets, J = 7, 2.5)

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] acetyl] amino 1.22 g of residue in -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt (first and second outputs), 4.2 ml of aniazole and trifluoro over 13 minutes at -15 ° C as described above. Treated with 16 ml roroacetic acid. Chromatography was carried out with a 300 ml HP-20AG column and the above treatment was carried out to obtain 694 mg of the target compound in fractions 6-9 (60 ml).

[Examples 104-133]

(Z) -2-amino-α-[[[[hydroxy (phenylmethoxy) phosphinyl] methoxy] imino-4-thiazoleacetic acid was used instead of the acid described in column I in the method of Example 11 Then, the compound described in column II was obtained.

[Examples 134 to 135]

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -2-oxoethoxy] imino] acetyl] amino] -2-oxo- The compound described in column II was obtained according to the method of Example 70 using 1-azetidinesulfonic acid, the compound described in column I, instead of the potassium salt.

Example 136

[3α (Z), 4α] -3-[[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

Soluble in (cis) -4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidisulfonic acid, 51.8 mg of potassium salt and 51 mg of tetra-n-butyl-ammonium disulfide The solution was extracted four times with 10 ml each of methylene chloride to obtain 81 mg of an oily substance. This material was stirred with 40 mg of activated carbon based 10% palladium in 4 ml of dimethylformamide under hydrogen atmosphere for 2 hours. The catalyst was filtered off and washed with 1 ml of dimethylformamide. Combine the filtrate and the wash and add 31 mg of (Z) -2-amino-α- (methoxyimino) -4-thiazole acetic acid, 27 mg of N-hydroxybenzotriazole and 31.5 mg of dicyclohexylcarbodiimide And stirred for about 16 hours. This solution was evaporated in vacuo and the residue was titrated with 3 ml of acetone. The resulting slurry was separated by centrifugation and treated with 51 mg of potassium perfluorobutanesulfonate. It was diluted with 5 ml of ether and filtered to give a solid. This was chromatographed with HP-20AG (40 ml) to obtain a lidon-positive substance in fractions 3-5 (20 ml) (eluted with water). The solvent was evaporated and titrated with ether to give 23 mg of the product in the form of an absorbent solid.

Analysis of C ₁₀ H ₁₂ N ₅ O ₆ S ₂ K

Theoretic value: C 29.91; H 3.01; Found: C 29.30; H 3.31;

N 17.44 N 16.66

NMR (D ₂ O)

1.40 (3H, d, J 7)

3.91 (3H, s)

4.46 (1H, Pentet J = 7)

5.37 (1H, doublet, J = 7)

6.97 ppm (1 H, s)

Example 137

(3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid

A) t-Boc-1-alorleonin

A suspension of 6.72 g of 1-allerleonin in 70 ml of 50% dioxane solution was treated with 9.45 ml of triethylamine and 18.1 g of t-butylpyrocarbonate. The mixture was stirred at rt for 4 h, diluted with 70 ml of water and then diluted with 140 ml of ethyl acetate. After shaking the mixture gently, the aqueous layer was separated, the organic layer was washed with 30 ml of water: salt (2: 1), and then combined to extract the aqueous layer again with 70 ml of ethyl acetate. The aqueous layer was cooled in an ice bath and the pH was adjusted to 2.3 by addition of 10% potassium disulfide solution. The acidified solution was extracted four times with 150 ml of ethyl acetate each time, and the combined organic layers were dried over anhydrous sodium sulfate, and the solvent was evaporated to remove 9.13 g of the target compound.

B) N-methoxy-t-boc-1-allothrooninamide

9.13 g of t-boc-1-allothroonine is dissolved in 85 ml and 4 ml of potassium hydroxide solution, and 5.22 g of methoxyamine hydrochloride and 8.67 g of 1-ethyl-3,3- (dimethylaminopropyl) carbodiimide HCl Was added. The mixture was stirred at rt for 4 h and then saturated with potassium tartrate. The resulting mixture was extracted four times with 150 ml of ethyl acetate each time, the organic layer was dried over anhydrous sodium sulfate, and the solvent was evaporated to remove 7.38 g of the target compound in the solid phase.

C) O-methanesulfonyl-N-methoxy-t-boc-1-allothroonineamide

7.32 g of N-methoxy-t-boc-1-allothroonine amide was dissolved in 40 ml of pyridine and cooled to −20 ° C. under a nitrogen atmosphere. 30 ml of methanesulfonyl chloride was added dropwise thereto over 5 minutes by syringe, and the resulting mixture was slowly warmed to 0 ° C. and stirred at this temperature for 3 hours. 500 ml of ethyl acetate was added thereto, and the solution was washed with 250 ml of 3NHCl solution cooled with ice, followed by 100 ml of 5% NaHCO ₃ solution. The ethyl acetate layer was dried over anhydrous sodium sulfate and the solvent was evaporated to yield 8.64 g of the desired compound as a white solid.

C) (3S-cis) -3-t-butoxycarbonylamino-1-methoxy-4-methylazetidinone

8.64 g of 0-methanesulfonyl-N-methoxy-t-boc-1-allorthreonine amide was dissolved in 530 ml of acetone, and 11 g of potassium carbonate solid was added thereto. The mixture was slowly heated to 65 ° C. in the presence of nitrogen and the mixture was stirred at this temperature for 1 hour. The reaction mixture was filtered through celite and the filter cake was washed with ethyl acetate. The residue obtained by concentrating the filtrate was dissolved in 250 ml of ethyl acetate. The ethyl acetate solution was washed with 100 ml of 1N hydrochloric acid solution and 100 ml of 9% sodium bicarbonate solution, the ethyl acetate layer was dried over anhydrous sodium sulfate, and the solvent was evaporated to give 6.63 g of crude product.

E) (3S-cis) -3-t-butoxycarbonylamino-4-methylazetidinone

1.35 g of sodium was dissolved in about 300 ml of ammonia water at -50 ° C, and dissolved in 35 ml of tetrahydrofuran (3S-cis) -3-t-butoxycarbonylamino-1-methoxy-4-methylazeti 5.87 g of dinone was added dropwise with a syringe. 10 ml of tetrahydrofuran was further added to wash. When the addition was almost complete, about 100 mg more sodium was added and the mixture was further stirred for 5 minutes, followed by quenching by adding 3.35 g of ammonium solids in one portion. Ammonia was blown off with a stream of nitrogen and 25 ml of ethyl acetate was added to the residue. After filtration, the solid was washed with ethyl acetate, the filtrates were combined, the solvent was evaporated to give 4.82 g of the target compound.

F) (3S-cis) -3-t-butoxycarbonylamino-4-methyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

4.98 g of [3S, 4R] -3-t-butoxycarbonylamino-4-methylazetidinone is dissolved in 30 ml of dimethylformamide, and 11.9 g of pyridine-sulfur trioxide complex is added thereto, and the mixture is nitrogen. Stir at room temperature in the presence. After stirring for 14 hours, 1.8 g of pyridine-sulfur trioxide complex was further added and stirred for 80 hours. The reaction mixture was poured into 700 ml of 0.5 M monobasic potassium phosphate solution, which was washed three times with 30 ml of methylene chloride. 8.45 g of tetra-n-butylammonium disulfide was added to the aqueous solution, and this was extracted four times with 300 ml of methylene chloride. The methylene chloride layer was collected, dried over anhydrous sodium sulfate, the solvent was removed, and 10.76 g of a rubbery target compound was obtained.

G) (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid

10.76 g of (3S-cis) -3-t-butoxycarbonylamino-4-methyl-2-oxo-1-azetidinesulfonic acid and tetrabutyl ammonium salt were dissolved in 50 ml of 95-97% formic acid, and in the presence of nitrogen, 4 Stir for hours. A small amount of product obtained in the previous step was added as a seed and the mixture was stirred for at least 1 hour. The mixture was stored in the freezer for about 16 hours and the frozen mixture was allowed to warm up to room temperature and then stirred for 1 hour more. The solid formed was filtered and then washed with methylene chloride to give 982 mg of the target compound. The filtrate was diluted with 1 L of methylene chloride and left for 4 hours at −20 ° C. The precipitate formed was recrystallized from water-methanol-acetone to give 167 mg of the desired compound.

NMR (D ₂ O)

1.63 (3H, doublet, J = 6.5 cps)

IR (new sol) 1,775cm ^-1

Example 138

[3- [3α (Z), 4α]]-3-[[2-amino-4-thiazolyl) -methoxyimino) acetyl] amino-4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

Dicyclohexylcarbodiimide 206 mg of a solution obtained by dissolving 201 mg of (Z) -2-amino-α- (methoxyimino-4-thiazole acetic acid and 153 mg of N-hydroxy-benzotriazole monohydrate in 3 ml of dimethylformamide And the mixture was stirred for 20 minutes at room temperature in the presence of nitrogen, followed by 180 mg of (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid and 0.14 ml of triethylamine Was added to a solution of 2 mg of dimethylformamide and washed with the addition of 1 ml of dimethylformamide The mixture was stirred for about 16 hours The slurry was evaporated in vacuo, titrated with 12 ml of acetone and then centrifuged. The liquid obtained by separation was treated with 338 mg of potassium perfluorobutanesulfonic acid, which was diluted with 10 ml of ether, filtered, and chromatographed with 200 ml of a solid product HP-20 resin obtained by filtration. After extraction, the fractions 18-30 (20 ml each) were collected and lyophilized to obtain 274 mg of the target compound of the absorbent solid.

Analysis of C ₁₀ H ₁₂ N ₅ O ₆ S ₂ K

Theoretic value: C 29.91; H 3.01; Found: C 30.03; H 3.21

N 17.44 N 17.06

NMR (D ₂ O)

1.40 (3H, d, J = 6.5) 5.36 (1H, d, J = 5.5)

3.98 (3H, s) 6.97 (1H, s)

4.48 (1H, dt, J = 6.4, 5.5)

Example 139

(3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid

A) threonine, methyl ester, hydrochloride

The flask containing 500 ml of methanol under nitrogen was cooled to −5 ° C. (ice / brine), and 130 ml (excess) of thionyl chloride was added thereto at a rate such that the temperature of the reaction was maintained between 0 ° C. and 10 ° C. After cooling back down to -5 [deg.] C., 59.5 g of 1-threonine were added and allowed to stand until the temperature of the mixture became room temperature and then stirred for 16 hours. The mixture was concentrated and vacuum evaporated at 10 ⁻¹ Torr for 2 hours to give a viscous oil. This material was used directly in the next step.

B) threonine amide

The crude product obtained in part A was dissolved in 2.5 L of methanol and cooled to -5 ° C (ice / brine). The solution was saturated with ammonia gas and then the cooling bath was removed and the reaction vessel was sealed and left for 3 days. Unreacted ammonia was removed with an aspirator, and then 100 g of sodium bicarbonate and 50 ml of water were added thereto, and the mixture was evaporated to concentration to give a viscous oil.

C) benzyloxycarbonylthreonine amide

The crude product obtained in part B (containing the required amount of sodium bicarbonate) is diluted with 1 liter of water, the solution is stirred quickly, and 94 g of benzyloxycarbonyl chloride (88 ml of 90% pure material) is added to 80 ml of tetrahydrofuran. The dissolved solution was added over 1 hour. The reaction mixture was further stirred for 16 h and then extracted with ethyl acetate (once with 500 ml, twice with 250 ml). The extracts were combined, dried over magnesium sulfate and then concentrated. The crystalline residue was dissolved in 250 ml of warmed ethyl acetate, 300 ml was added to it and then boiled until a clear solution. After cooling, the resulting crystalline material was filtered and dried to give 104 g of the target compound.

D) benzyloxycarbonylthreonineamide, O-mesylate

Under argon atmosphere, 100 g of benzyloxycarbonylthreonine amide was dissolved in 400 ml of anhydrous pyridine and cooled in an ice / salt bath. While stirring this solution, 36.8 ml (54.5 g) of methanesulfonyl chloride was added thereto over 15 minutes, further stirred for 2 hours, and then 0.3 equivalent of methanesulfonyl chloride was further added thereto. The reaction mixture was stirred for 1 hour and then poured into a mixture of 1.5 L ice and 2 L water. The resulting slurry was stirred for about 30 minutes and then filtered. The crude product was dried in a vacuum oven at 60 ° C. for 16 hours to afford the desired compound 109.

E) N-sulfonyl benzyloxycarbonylthreonineamide, O-mesylate, tetrabutylammonium salt

A solution of 17.8 ml of 2-picoline in 90 ml of methylene chloride was cooled to -5 deg. C (ice-brine), and 5.97 ml of chlorosulfonic acid were added thereto at a rate capable of keeping the internal reaction temperature below 5 deg. . The resulting solution was added to a 7.56 g suspension of benzyloxycarbonylthreonine amide, O-mesylate suspended in 120 ml of methylene chloride via cannula. The resulting heterogeneous mixture was refluxed for about 16 hours to give a clear solution. This solution was poured into 500 ml of 0.5 M phosphate buffer at pH 4.5 and further diluted with 120 ml of methylene chloride. The organic layer was separated, washed once with 100 ml of buffer, the aqueous layer was collected, treated with 10.2 g of tetra-n-butylammonium hydrogen sulfate, and extracted with methylene chloride (once with 300 ml and twice with 150 ml each). The organic extracts were combined, dried over sodium sulfate and the solution was concentrated to give 127 g of foamy material.

F) (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid

A mixture of 5.52 g of potassium carbonate dissolved in 20 ml of water and 160 ml of 1,2-dichloroethane was refluxed, and N-sulfonyl benzyloxycarbonylthreonine amide in 20 ml of 1,2-dichloroethane (20 ml for washing) was added thereto. , O-mesylate and 15.5 mmol of tetrabutylammonium salt were added. After refluxing for 30 minutes, the mixture was poured into a separatory funnel, diluted with 50 ml of water and 100 ml of methylene chloride, and the phases were separated. The resulting organic layer was dried over sodium sulfate and concentrated to give crude (3S-trans) -3-benzyl oxycarbonylamino-4-methyl-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt. Zoazetidinone was treated with 0.8 g of an activated carbon based 5% palladium catalyst in 250 ml of ethanol, and hydrogen was injected into the solution while generating bubbles. After 90 minutes the mixture was filtered through celite and washed with 50 ml of ethanol. 1.2 ml of formic acid was added to the solution to immediately precipitate the desired zwitterionic material, which was stirred for 1 hour, then filtered and dried at 10-11 lots for 1 hour to obtain 1.1 g of product. The filtrate was concentrated and then further formic acid was added to give 1.3 g of the desired zwitterionic material as a second output. Melting point 218 ° C. or higher (decomposition).

[α] ^D = -41.4 (c = 1, H ₂ O)

NMR (D ₂ O) 1.58 (3H, d, J = 7)

4.80 (2H, M)

[Examples 140 to 143]

(3S-trans) -3-amino-4-methyl-2-oxo-1-azetidyl sulfonic acid is used instead of (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid And using the acid described in Column I in place of (Z) -2-amino-α- (methoxyimino) -4-thiazoleacetic acid and following the method of Example 138 to obtain the compound described in Column II.

Example 144

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [[1,1-dimethyl-2-[(4-nitrophenyl) methoxy] -2- Oxoethoxy] imino] acetyl] amino] -4-petyl-2-oxo-1-azetidinesulfone, potassium salt

309 μl of triethylamine in 0.36 g of a slurry of (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (see Example 139) in 30 ml of anhydrous dimethylformamide was nitrogen atmosphere at 26 캜. Under, and after about 5 minutes a clear solution was obtained. 0.816 g of (Z) -2-amino-α-[[1,1-dimethyl-2-[(4-nitrophenyl) methoxy] -2-oxoethoxy] imino] -4-thiazole acetic acid After the addition, 0.334 g of N-hydroxybenzotriazole and 0.453 g of dicyclohexylcarbodiimide were added. The mixture was stirred at 26 ° C. for 12 hours, then the solvent was removed in vacuo and the residue was titrated with 30 ml of acetone. After stirring for 5 minutes, the solids were removed and the filtrate was treated with 3.680 g of potassium sulfate perfluorobutane dissolved in 5 ml of acetone. About 40 ml of ether was added thereto, and the precipitated material was collected and dried in vacuo to give 1.073 g of the first output, followed by 0.066 g of the output to obtain 1.14 g of all.

Analysis of C ₂₀ H ₂₁ N ₆ O ₁₀ S ₂ K · 1H ₂ O

Theoretic value: C 38.33; H 3.70; Found: C 38.30; H 3.63;

N 13.41; S 10.23; N 13.41; S 9.88;

K 6.25 K 5.98

Example 145

[3α (Z), 4α] -3-[[(2-amino-4-thiazolyl) ([1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl-2-oxo -1-azetidinesulfonic acid, potassium salt (1: 2)

A) [3α (Z), 4α] -3-[[2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] acetyl] amino] -4 -Methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

(Cis) -4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, tetrabutylammonium salt (as described in Example 136, corresponding to Example 98) 201 mg of dimethylformamide was dissolved in 5 ml of 10% palladium based on calcium carbonate under a water atmosphere for 2 hours. The slurry was filtered, and the filtrate example (Z) -2-amino-α- [1-diphenylmethoxycarbonyl-1-methylethoxy) imino) -4-thiazole acetic acid 146 mg, dicyclohexabodiimide 73 mg and 51 mg of N--hydroxybenzotriazole were added and then stirred for about 16 hours under a nitrogen atmosphere. The slurry was evaporated in vacuo and titrated with 4 ml of acetone. The solid water obtained by filtration of the slurry was washed twice with 2 ml of acetone, and the filtrate and wash were combined and treated with 113 mg of potassium sulfate perfluorobutane. The solid obtained by diluting it with 24 ml of ether was isolated by centrifugation and washed three times with ether to obtain 186 mg of the target compound.

B) [3α (Z), 4α] -3-[[(2-amino-4-thiazolyl) [1-carboxy-1-methylethoxy] imino] acetyl] amino] -2-methyl-4- Oxo-1-azetidinesulfonic acid, potassium salt (1: 2)

[3α (Z), 4α] -3-[[2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino) acetyl] amino] -4-methyl The slurry dissolved in 0.6 ml of 2-oxo-1-azetidinesulfonic acid and 186 mg of potassium salt distilled was cooled to -12 ° C, and 3.0 ml of difluorotrifluoroacetic acid was added thereto at -10 ° C. . The solution was stirred for 10 minutes, 12 ml of ether was added, followed by 6 ml of hexane. After 5 minutes at −10 ° C., stirring at ambient temperature for 15 minutes, the solids were isolated by centrifugation, washed four times with ether, and 141 mg of the material obtained by drying in vacuo, then obtained in powder form, It was dissolved in 5 ml of cold water and then adjusted to pH5.6 by immediate addition of 0.4N potassium hydroxide solution. This solution was eluted with water using a 100 ml HP-20AG column. Fractions 8-12 (10 ml) were combined and evaporated in vacuo (acetonitrile was added three times followed by evaporation). The residue was ether titrated to give 101.7 mg of absorbent solid.

Analysis of C ₁₃ H ₁₅ N ₅ O ₈ S ₂ · 2K

Theoretic value: C 30.51; H 2.95; Found: C 30.11; H 3.26;

N 13.69; S 12.53; N 13.35; S 12.12;

K 15.28 K 15.02

Example 146

[3S- [3α (Z), 4β]]-3 [[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl- 2-oxo-1-azetidinesulfonic acid

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl)-[(1-carboxy-1-methylethoxy] imino] acetyl] amino] -4- 87.3 mg of methyl-2-oxo-1-azetidinesulfonic acid, dipotassium salt (see Example 103) are dissolved in 1.38 ml of water, cooled to 0 ° C., and treated with 0.34 ml of 1N hydrochloric acid to centrifuge the crystals produced. The wet solid was dissolved in methanol, filtered, concentrated to about 0.5 ml and mixed with 1 ml of water to give 55.9 mg of the desired compound.

Example 147

[3S- [3α (Z) -4β]]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] aceral] amino] -4- Methyl-2-oxo-1-azetidinesulfonic acid, sodium salt

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl 99.7 mg of 2-oxo-1-azetidinesulfonic acid were mixed with 0.207 ml of 1N sodium hydroxide, and the mixture was slowly warmed to dissolve the remaining solid. Water was removed by azeotroping with acetonitrile and the residue was crystallized from a mixture of 0.5 ml of methanol (to dissolve the residue) and 1 ml of acetonitrile to give 81.8 mg of a solid. Second recrystallization with 0.8 ml of methanol gave 47.9 mg of 47.9 mg, 0.24 ml of methanol and 0.24 ml of anhydrous methanol, and 38.8 mg of the fourth with 0.225 ml of ethanol and 0.225 ml of anhydrous ethanol. This solid was dried at 20 ° C. and 0.01 mg Hg for 18 hours and then equilibrated with atmospheric moisture for 24 hours to give 40.9 mg of the target compound.

Example 148

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl 2-oxo-1-azetidinesulfonic acid, disodium salt

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl)-[(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4- 3.00 g of methyl-2-oxo-1-azetidinesulfonic acid (see Example 146) was suspended in 30 mg of water, followed by titration with 12.0 ml of 1N sodium hydroxide to obtain the disodium salt of the target compound. A small amount of Dowex 50W-X2 (H ⁺ ) was added to lower the pH to 6.5. The mixture was filtered and the filtrate was diluted with water to give 66.3 g. 6.63 g was separated and used for other purposes. The remaining filtrate was lyophilized to give 2.38 g of a solid. Partially equilibrated with moisture in the air for 24 hours to obtain 2.54 g of the target compound.

[Examples 149-151]

The compound described in Column I was used in accordance with the method of Example 138, using the compound described in Column I in place of (Z) -2-amino-α- (methoxyimino) -4-thiazole acetic acid.

Example 152

[3S- [3α (Z), 4α]]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-methyl 2-oxo-1-azetidinesulfonic acid, potassium salt (1: 2)

A) [3S- [3α (Z), 4α]]-3-[[(2-amino-4-thiazolyl)-[(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] Acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

440 mg of (Z) -2-amino-α-[(1-carboxy-1-methylethoxy) imino] -4-thiazole acetic acid and 153 mg of N-hydroxybenzotriazole monohydrate were dissolved in 3 ml of dimethylformamide. The resulting solution was treated with 206 mg of dicyclohexylcarbodiimide. This mixture was plated for 30 minutes at room temperature in the presence of nitrogen, to which (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (see Example 137) and triethylamine A solution of 0.14 ml dissolved in 2 ml of dimethylformamide was added (washed with an additional 1 ml of dimethylformamide) and the mixture was plated for about 16 hours. The slurry was evaporated in vacuo and titrated with 12 ml of acetone. The solid obtained by filtration of the slurry was washed twice with 3 ml of acetone, and the filtrate and washings were combined and treated with 338 mg of potassium sulfate perfluorobutane. The rubbery solid obtained by diluting this with 30 ml of ether was slowly solidified. This solid was filtered and washed with ether to give 656 mg of the target compound.

B) [3S- [3α (Z) -4α]]-3-[[(2-amino-4-thiazole) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4 Methyl-2-oxo-1-azetidinesulfonic acid, potassium salt (1: 2)

[3S- [3α (Z), 4α]]-3 [[(2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] acetyl] amino] Slurry of 4-methyl-2-oxo-1-azetidinesulfonic acid and potassium salt 656 mg in 2.3 ml of distilled anisole was cooled to -12 ° C, and trifluoroacetic acid (precooled to -10 ° C) was added. 11.5 ml of Sikkim were added, the solution was stirred for 15 minutes, to which 46 ml of ether followed by 23 ml of hexane, held at −10 ° C. for 5 minutes, then stirred at room temperature for 15 minutes, and then the resulting solid phase Water was filtered and washed with ether to give 457 mg of a very absorbent gummy material, which was dissolved in 6 ml of cold water and immediately added 0.4N potassium hydroxide solution to adjust the pH to 5.6. Eluted with water using HP-20 resin, fractions 7-11 (50 ml each) were combined and lyophilized to form a solid. 239 mg of the mixture was obtained.

Analysis of C ₁₃ H ₁₅ O ₉ N ₅ S ₂ K ₂ · ½H ₂ O

Theoretical: C 29.99; H 3.10; Found: C 29.94; H 3.30;

N 13.45; S 12.32 N 13.30; S 11.93

NMR (D ₂ O)

1.44 (3H, d, J = 75) 5.34 (1H, d, J = 5.5)

1.46 (6H, s) 6.96 ppm (1H, s)

4.48 (1H, dt, J = 75, 5.5)

Example 153

(±) -3-amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid

A) (±) -4,4-dimethyl-2-oxo-1-azetidine-t-butyldiphenylsilane

A solution obtained by dissolving 40.5 ml of t-butylchlorodiphenylsilane in 112 ml of dimethylformamide was cooled to 0 ° C., and 22 ml of triethyl amine was added thereto. A solution in which 12.87 g of 4,4-dimethyl-2-azetidinone was dissolved in 25 ml of dimethylformamide was added dropwise to the cooled triethylamine solution over 10 minutes. The resulting cloudy solution was stirred for 18 h at 5 ° C. in the presence of argon, and then the mixture was poured into 400 ml of ice water and extracted three times with 150 ml of ether: ethyl acetate (2: 1). The combined extracts were washed four times with 100 ml of 0.5 M monobasic potassium phosphate buffer, once with 150 ml of sodium bicarbonate solution, twice with 150 ml of water, and once with 150 ml of saturated sodium chloride solution. The solution was dried over sodium sulfate and concentrated in vacuo to give 33.0 g of the solid target compound.

B) (±) -3-azide-4,4-dimethyl-2-oxo-1-azetidine-t-butyldiphenylsilane

A solution of 4.25 ml of 1.6 Mt-butyllilium dissolved in hexane and 11 ml of anhydrous tetrahydrofuran was prepared at -50 ° C in the presence of argon in a three-necked flask for 100 ml. To this was added a solution of 0.083 g of triphenylmethane dissolved in 1 ml of tetrahydrofuran, the resulting solution was cooled to -60 ° C, and 1.0 ml of diisopropylamine was added dropwise thereto by syringe. The solution was stirred for 15 minutes and then cooled to -78 ° C. To this, a solution of 2.3 g of (±) -4,4-dimethyl-2-oxo-1-azetidine-t-butyldiphenylsilane dissolved in 8 ml of tetrahydrofuran was slowly added dropwise to the syringe. While the resulting solution was stirred at −78 ° C. for 20 minutes, severe precipitation occurred, making it difficult to stir evenly. To this, a solution of 1.33 g of p-toluenesulfonyl azide dissolved in 5 ml of tetrahydrofuran was added dropwise, stirred at −78 ° C. for 20 minutes, and then 2 ml of trimethylsilyl chloride was added dropwise. The reaction mixture was allowed to warm to ambient temperature and then stirred for 1 hour. The mixture was cooled to 0 ° C. and poured into 150 ml of ethyl acetate (0 ° C.). Sufficient 0.5M monobasic phosphate buffer was added to clear the aqueous and organic layers. The two layers were separated, and the organic layer was washed three times with 150 ml of 0.5 M monobasic potassium phosphate solution, once with 150 ml of sodium chloride solution and once with 150 ml of saturated sodium chloride solution, and then dried over sodium sulfate. 2.83 g of the oily substance obtained by concentrating this solution in vacuo was titrated with hexane to obtain 1.67 g of the target compound in the solid phase.

C) (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidine

In a 50 ml three-necked flask, 1.52 g of (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidine-t-butyldiphenylsilane was dissolved in 25 ml of acetonitrile. The solution was stirred and 0.25 ml of 48% hydrofluoric acid was added thereto. The solution was stirred at ambient temperature, to which a total of 3.25 ml of 48% hydrofluoric acid was added, with 48% hydrofluoric acid added 0.5 ml every 60 minutes for 6.5 hours. The reaction mixture was cooled to 0 ° C., neutralized with saturated sodium bicarbonate solution, and extracted with 120 ml of ethyl acetate. The organic layer was then washed with 100 ml of water and 100 ml of saturated sodium chloride solution and then dehydrated with sodium sulfate. This dehydration solution was concentrated in vacuo to give 1.34 g of an oily substance. The oily substance containing this impurity was chromatographed with 27 g of silica gel, eluted with tohexane and then with 33% ethyl acetate in hexane to obtain 0.358 g of the target compound in the solid phase.

D) (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

To 0.100 g of (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidine 2.8 ml of 0.5 M dimethyl formamide-trioxide sulfur complex was added in the presence of argon at 0 ° C. and the mixture was Warm to ambient temperature and then stir for 45 minutes. This solution was then poured into 20 ml of 0.5 M monobasic potassium phosphate buffer (pH5.5), washed three times with 20 ml of methylene chloride, and 0.237 g of hydrogen tetrabutylammonium sulfate was added to this aqueous solution. This was extracted four times with 20 ml of methylene chloride and the combined organic extracts were washed with 20 ml of 8% sodium chloride solution. The methylene chloride solution was dehydrated with sodium sulfate and concentrated in vacuo to yield 0.31 g of an oily substance, which was analyzed by NMR to give 50% of dimethylformamide and 50% of the target compound.

E) (±) -3-amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid

0.155 g of (±) -3-azido-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid tetrabutylammonium salt was dissolved in 0.6 ml of methanone and activated for 20 minutes at 1 atmosphere in the presence of 10% palladium based on activated carbon. Hydrogenated. The catalyst was filtered off, washed with methylene chloride and combined with methanol solution. This clear solution was treated with 0.123 ml of 97% formic acid. After acid addition, the solution was immediately clouded. After leaving this solution at 5 ° C for 1 hour, the solid was filtered to obtain 0.0664 g of the target compound having a melting point of 200 ° to 202 ° C (decomposition).

NMR (D ₂ O)

Example 154

[3 ± (Z)]-3-]] (2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4,4-dimethyl-2-oxo-1-azetidinesulfonic acid, potassium salt

50 mg of N-hydroxybenzothiazole hydrate and 0.323 mmol of (Z) -2-amino-α- (methoxyimino) -4-thiazole acetate were dissolved in 0.5 ml of dimethylformamide at ambient temperature in the presence of argon. Treatment with dicyclohexylcarbodiimide 67 mg. The mixture was stirred for 1 hour, to which 57 mg of (±) -3-amino-4,4-dimethyl-2-oxo-1-azetidinesulfonic acid (see Example 153) was added in the form of a solid, Then 0.05 ml of triethylamine was added dropwise. The reaction was stirred at ambient temperature for 16 hours. Dimethylformamide was removed at 30 ° C. and under high vacuum, and the residue was slurried with 4 ml of acetone and filtered. The filter cake was washed with 4 ml of acetone, and 85 mg of potassium sulfate perfluorobutane was added to the filtrate, followed by ether. The resulting rubbery substance was titrated with ether to give 40 mg of a tan solid, which was chromatographed with a 70 ml HP-20AG column, eluted with water, and fractions 16-40 (5 ml) were evaporated. Obtained and titrated with acetone-hexane (1: 1) and dried. Melting point 225 ° C. (decomposition).

Analysis for _{C 11 H 14 N 5 S 2} · K

Theoretic value: C 31.80; H 3.40 Found: C 29.47; H 3.48;

N 16.86; S 15.43 N 14.98; S 13.35

Example 155

(±) -4,4-dimethyl-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

To a solution of 45 mg of N-hydroxybenzotriazole hydrate and 40 mg of phenylacetic acid dissolved in 0.5 ml of dimethylformamide was added 61 mg of dicyclohexylcarbodiimide at ambient temperature in the presence of argon. The mixture is stirred for 1 hour, to which 52 mg of (±) -3-amino-4,4-dimethyl-2-oxo-1-acetidinesulfonic acid (see Example 153) is added in the solid state, Then 0.04 ml of triethylamine was added dropwise. The reaction was stirred at ambient temperature for 24 hours and then dimethylformamide was removed at high vacuum and 30 ° C., and the residue slurried in acetone and filtered. Potassium sulfate perfluorobutane was added to the filtrate, ether was added, and the mixture was cooled. The resulting solid was washed with acetone and hexane and then dried to obtain the target compound in powder form.

Analysis of C ₁₃ H ₁₅ N ₂ O ₅ SK

Theoretic value: C 44.55; H 4.32; Found: C 43.83; H 4.16

N 8.00; S 9.15 N 7.96; S 8.76;

K 11.16; K 11.43

NMR (D ₂ O)

1.33 (s, 3H) 4.70 (s, 1H)

1.58 (s, 3H) 7.56 ppm (broad s, 5H)

3.68 (s, 3H)

Example 156

(3S-trans) -3-[[(2-amino-4-thiazolyl) oxoacetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

To a solution of 1.85 g of diphenylphosphinyl chloride dissolved in 15 ml of anhydrous dimethylformamide cooled in an ice-methanol bath (-15 DEG C to -20 DEG C), (2-amino-4-thiazolyl) glyoxalic acid and triethylamine 2.14 g of salt was added. After stirring for 0.5 hour, 1.08 g of (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidine sulfonic acid (see Example 139) and 1.92 ml of triethylamine were added to 5 ml of anhydrous dimethylformamide. The solution dissolved in was added to the cooled mixed anhydride solution and the reaction mixture was stirred at 5 ° C. for 24 hours. The solvent was removed in vacuo, the dark oily residue was dissolved in water, chromatographed with 200 ml of Dowex 50 × 2-400 resin (K ⁺ type), eluted with water (15 ml of fractions), and fraction 13-. 3.37 g of crude product was obtained at 27. Chromatography with HP-20 resin (200 ml) and eluting with water (15 ml fractions) afforded the desired compound in fractions 18-26. Water was removed in vacuo to obtain an amorphous powdery target compound.

Analysis of C ₉ H ₉ N ₄ O ₆ S ₂ K (372.42)

Theoretic value: C 29.02; H 2.44; Found: C 28.87; H 2.62;

N 15.04; S 17.22 N 14.85; S 15.09;

K 10.50 K 10.81

Example 157

[3S (R ^* )]-3-[[[(aminoacetyl) amino] phenylacetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

Trifluoroacetate (1: 1)

[3S (R ^* )]-3-[[[[[[(4-methoxyphenyl) methoxy] carbonyl] amino] acetyl] amino] phenylacetyl] amino using trifluoroacetic acid and aniazole ] -2-oxo-1-acetidine sulfonic acid potassium salt (refer Example 172) was removed, and the target compound of melting | fusing point 165 degreeC (decomposition) was obtained.

Example 158

(3S-trans) -3-methoxy-4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

A) (3S-trans) -4-methyl-3-methoxy-2-oxo-4-[[(phenylmethoxy) carbonyl] amino) azetidine]

(3R-trans) -4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] azetidine (yield from d-threonine as described for racemicis isomers in Example 98C) 126%) was dissolved in 112 ml of 4% Borax in methanone and cooled to 0 ° C., to which 3.5 ml of t-butyl hypochloride was added. After 20 minutes, the solution was poured into 1 liter of cold water, extracted twice with 750 ml of cold ethyl acetate, and then the organic layer was washed twice with 750 ml of cold water, washed with saturated salt, dried and evaporated to give crude N, 3.05 g of N'-dichloroamide was obtained.

A solution of 426 mg of lithium methoxide dissolved in 20 ml of anhydrous methanone was cooled to -78 ° C, which was diluted with 40 ml of anhydrous tetrahydrofuran. The solution of the chloroamide dissolved in 20 ml of tetrahydrofuran at -78 ° C was added via syringe over 30 seconds. After 20 minutes at −78 ° C., acetic acid and trimethyl phosphite were added 2 ml each. After maintaining at room temperature for 40 minutes, the solution was poured into 500 ml of water and extracted twice with 300 ml of ethyl acetate. The organic layer was washed with water, dried and evaporated to give an oily substance. Chromatography was carried out on a 200 ml silica gel column, and eluted with chloroform-ethyl acetate (3: 1) to obtain 1.25 g of the target compound.

B) (3S-trans) -3-methoxy-4-methyl-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidinesulfonic acid, potassium salt

A solution of 800 mg (0.00303 mol) of (3S-trans) -4-methyl-3-methoxy-2-oxo-4-[[(phenylmethoxy) -carbonyl] amino] azetidine in 2 ml of dimethylformamide Was cooled to 0 ° C., and 4 ml of dimethylformamide-sulfur trioxide complex was added thereto. After holding at 0 ° C. for 1 hour and at room temperature for 4 hours, the solution was poured into 80 ml of 0.5 M monobasic potassium phosphate (adjusted to pH 5.5) and extracted twice with 50 ml of methylene chloride. The aqueous layer was treated with 1.04 g of tetrabutylammonium sulfate and then extracted with dichloromethane to give 1.42 g of an oily substance. This material was dissolved in acetone and treated with 0.04 g of potassium sulfate perfluorobutane in 10 ml of acetone. This was diluted with 250 ml of ether and the oily solid was titrated to give 584 mg of crude product. Chromatography with HP-20AG (200 ml) eluted with 1 liter of water followed by water-acetone (9: 1) to give 418 mg of purified product from fractions 13-16 (100 ml). This material, 114 mg, was titrated with ether to give 104 mg of analytical sample.

Analysis of C ₁₃ H ₁₄ N ₂ O ₇ SK · H ₂ O

Theoretic value: C 39.06; H 4.04 Found: C 38.91; H 3.62;

N 7.01; S 8.03; N 6.91; S 8.06

K 9.78 K 9.51

NMR (D ₂ O)

1.33 (3H, d, J = 7) 5.18 (2H, s)

3.46 (3H, s) 7.43ppm (5H, s)

4.22 (2H, dd, J = 6)

Example 159

(3S-trans) -3-methoxy-4-methyl-2-oxo-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

(3S-trans-3-methoxy-4-methyl-2-oxo-3-[[(phenylmethoxy) -carbonyl] amino] -1-azetidinesulfonic acid, potassium salt (see Example 158) (3S-trans) -3-amino-3-methoxy-4-methyl-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salts were prepared by converting to a thiammonium salt and then adding a hydrogenation catalyst. The procedure of Example 88 was repeated except that trans) -3-amino-3-methoxy-4-methyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt, and phenylacetyl chloride were used. The compound was obtained.

Analysis of C ₁₃ H ₁₅ N ₂ O ₆ SK

Theoretic value: C 42.61; H 4.31 Found: C 39.67; H 4.09;

N 7.65 N 7.30

NMR (D ₂₀ )

1.29 (BH, d, J = 7) 4.46 (2H, dd, J = 6)

3.45 (3H, s) 7.38 ppm (5H, s)

3.73 (2H, s)

Example 160

[3S- (3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -2-oxoethoxy] imino] acetyl] amino ] -2-methyl-4-oxo-1-acetidinesulfonic acid, potassium salt

(Z) -2-amino-α-[[2- (diphenylmethoxy) -2-oxoethoxy] instead of (Z) -2-amino-α- (methoxyamino) -4-thiazole acetic acid Amino] -4-thiazole acetic acid was used, except that initially (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid was treated with triethyl amine. It carried out according to the method of Example 138, and obtained the target compound of melting | fusing point 155 degreeC-160 degreeC (decomposition).

Example 161

[3S- [3α (Z), 4β]]-3-[[[((carboxymethoxy) imino] (2-amino-4-thiazolyl) acetyl] amino] -4-methyl-2-oxo-1 Azetidinesulfonic acid, dipotassium salt

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -2- using trifluoroacetic acid and anisole The desired compound was decomposed at 250 ° C. or higher by removing a protecting group from oxoethoxy] imino] acetyl] amino-2-methyl-4-oxo-1-azetidinesulfonic acid potassium salt (see Example 160).

Example 162

(S) -3-[[[((2,6-dichloro-4-pyridinyl) thio] acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

(S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) to [(2,6-dichloro-4-pyridinyl) rio] acetic acid, 4-nitrophenyl ester The mixture was acylated with potassium sulfate, and then treated with potassium sulfate perfluorobutane to obtain a target compound having a melting point of 212 to 214 캜.

Example 163

[3S (R ^* )]-3-[[[[(4-amino-2,3-dioxo-1-piperazinyl) -carbonyl] amino) phenylacetyl] amino] -4-methyl-2- Oxo-1-azetidinesulfonic acid, potassium salt

[3S (R ^* )]-[4 [[[2-[(4-methyl-2-oxo-1-sulfo-3-azetidinyl) amino] -2-oxo-1-phenylethyl] amino] carr Carbonyl] -2,3-dioxo-1-piperazinyl] carnamic acid, phenylmethyl ester, potassium salt (see Example 149), using 10% palladium based on activated carbon as a catalyst and hydrogen gas It added and obtained the target compound of melting | fusing point 165 degreeC (decomposition).

Example 164

[3S (Z)]-3-[[(2-amino-4-thiazolyl)] [(1-carboxy-1-methylethoxy) imino) acetyl] amino) -2-oxo-1-azetidine Sulfonic acid, sodium salt (1: 2)

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] imino] acetyl] amino ] -2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt (see Example 28) were removed using trifluoroacetic acid and anisol to remove the protecting group, and then, an aqueous sodium hydroxide solution was added to the sodium salt. After conversion, purification with HP-20 afforded the target compound at melting point of 185 ° C. (decomposition).

[Examples 165 to 168]

(Z) -2-amino-α-[[[[hydroxy (phenylmethoxy) phosphinyl] methoxy] iminothi] -4-azolacetic acid is used instead of the acid described in column I in the method of Example II Then, the compound described in column II was obtained.

Example 169

(Trans) -3-amino-4-ethyl-2-oxo-1-azetidinesulfonic acid

A) t-Boc-N-methoxy-β-threoethylserineamide

1.33 g of threo-D, L-β-ethylserine was dissolved in 10 ml of 2N potassium hydroxide and 5 ml of t-butanol, to which 2.46 g of di-t-butylpyrocarbonate was added, followed by a biphasic mixture. Was stirred at ambient temperature for 4 hours. 1.25 g of 0-methylhydroxyammonium chloride was added thereto, followed by addition of 1N hydrochloric acid to adjust the pH to 4. 1.92 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide hydrochloride was added thereto, and pH was adjusted to 4 again. After stirring for 1 hour, the reaction mixture was saturated with sodium chloride and extracted four times with 50 ml of ethyl acetate. The ethyl acetate extracts were combined and dried over MgSO ₄ . The solution was raised in vacuo to obtain 1 g of the target compound.

B) t-Boc-O-methanesulfonyl-N-methoxy-β-thropionamide

10.5 g of t-Boc-N-methoxy-β-threoethylserineamide was dissolved in 65 ml of pyridine, and 4.65 ml of methanesulfonyl chloride was added dropwise at 0 ° C. After stirring for 3 hours at ambient temperature, the reaction mixture was poured into 200 g of ice and 300 ml of 1N hydrochloric acid. Concentrated hydrochloric acid was added to adjust the pH to 4, extracted three times with 85 ml of ethyl acetate, and the combined extracts were carbohydrated with MgSO ₄ and concentrated in vacuo. The residue was treated with carbon tetrachloride and then concentrated again. Ether was added and stirred, followed by filtration to give 6.9 g of the target compound.

C) (trans) -3-t-butoxycarbonylamino-4-ethyl-1-methoxy-2-azetidinone

4.15 g of anhydrous potassium carbonate and 125 ml of anhydrous acetone were refluxed, and 3.4 g of t-Boc-O-methanesulfonyl-N-methoxy-β-treopionamide dissolved in 25 ml of acetone was added thereto. After 1 h, the reaction mixture was cooled down, filtered and the filtrate was concentrated in vacuo. Hexane was added to the oily residue and stirred to obtain 2.2 g of the target compound.

D) (trans) -3-t-butoxycarbonylamino-4-ethyl-2-azetidinone

3 g of (trans) -3-t-butoxycardylamino-4-ethyl-1-methoxy-2-azetidinone was added to 170 ml of ammonia water at -78 ° C under nitrogen atmosphere, and 1.68 g of sodium was added in 5 parts. The mixture was stirred and added over 5 minutes. After stirring for 30 minutes, ammonium chloride was slowly added until the blue color of the reaction mixture disappeared. Ammonia was removed under nitrogen atmosphere, and the solid was extracted twice with 100 ml of ethyl acetate. The solvent was removed and then dried in vacuo to yield 2.7 g of the target compound.

E) (trans) -3-t-butoxycarbonylamino-4-ethyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

3.7 ml of trimethylsilylsulfonyl chloride dissolved in 5 ml of anhydrous dichloromethane was added to 2 ml of anhydrous pyridine dissolved in 20 ml of anhydrous dichloromethane. This addition was performed over 10 minutes in -30 degreeC and nitrogen presence. After stirring for 30 minutes at ambient temperature, pyridine-sulfur trioxide complex was obtained in a flask. (2.67 g of trans-3-t-butoxycarbonylamino-4-ethyl-2-azetidinone and 20 ml of anhydrous pyridine were placed in a flask and the flask was placed in an oil bath preheated to 90 ° C. After 15 minutes, A clear solution was obtained, which was poured into 200 ml of 1 M potassium dibasic phosphate solution, and 27 g of potassium dibasic phosphate and 100 ml of water were added to obtain a clear solution, which was extracted twice with 60 ml of ethyl acetate. Hydrogen tetrabutylammonium sulfate was added, and this aqueous solution was extracted three times with 100 ml of dichloromethane, and then the organic layers were combined and dehydrated with MgSO ₄ .

Concentration in vacuo afforded 6.9 g of the target compound.

F) (trans) -3-amino-4-ethyl-2-oxo-1-acetidinesulfonic acid

6.75 g of (trans) -3-t-butoxycarbonylamino-4-ethyl-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt were added to 40 ml of 98% formic acid and stirred at ambient temperature for 3 hours. . To this was added 60 ml of dichloromethane and the mixture was frozen for about 16 hours. The resulting precipitate was filtered off and dried in vacuo to yield 0.85 g of the target compound at a melting point of 185 ° C. (decomposition).

Example 170

(Trans Z) -3-[[(2-amino-4-thiazolyl [(1-carboxy-1-methylethoxy) imino] acetyl] amino-4-ethyl-2-oxo-1-azetidinesulfonic acid Dipotassium salt

A) (trans, Z) -3-[[(2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] acetyl] amino] -4-ethyl 2-oxo-1-azetidinesulfonic acid, dipotassium salt

0.55 g of (trans) -3-amino-4-ethyl-2-oxo-1-azetidinesulfonic acid and 335 mg of triethylamine are dissolved in 50 ml of anhydrous dimethylformamide, and (Z) -2-amino-α- 1.14 g of [(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] -4-thiazole acetic acid was added with stirring at 0 ° C., followed by 450 mg of hydroxybenzotriazole and dicyclohexylcar 0.69 g of bodyimide was added. After stirring at 0 ° C. for about 16 hours, the flask was emptied. 25 ml of anhydrous acetone was stirred and added to the obtained solid. The mixture was filtered and 0.94 g of potassium sulfate perfluorobutane was added to the filtrate, followed by 100 ml of ether. After standing at 0 ° C. for 1 hour, the solid was filtered off, washed with ether and dried in vacuo to yield 1.58 g of the target compound.

B) (trans, Z) -3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-ethyl-2-oxo- 1-azetidinesulfonic acid, dipotassium salt

(Trans, Z) -3-[[(2-amino-4-thiazolyl)] (1-diphenylmethoxycarbonyl-1-methylethoxy) imino] acetyl] amino suspended in 10 ml of anisol To a suspension of 1.31 g of 4-ethyl-2-oxo-1-azetidinesulfonic acid and dipotassium salt, 5 ml of trifluoroacetic acid was added at -15 占 폚 for 10 minutes. After stirring for 2 h at -10 ° C, a clear solution was obtained. 80 ml of anhydrous ether was added thereto at -30 ° C, and the resulting precipitate was filtered and then treated with 5 ml of water. 1N potassium hydroxide was added at 0 ° C. to adjust the pH to 5.5, and then the mixture was filtered to remove unconverted starting material. The filtrate was chromatographed with HP-20 using water as eluent, and then freeze-dried to give 185 mg of the target compound having a melting point of 160 ° C. (decomposition).

Example 171

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [(4-hydroxy-4-oxobutoxy) imino] acetyl] amino) -2-oxo-1-azetidine Sulfonic acid, potassium salt

[3S (Z)]-3-[[(2-amino-4-thiazolyl) [[4- (diphenylmethoxy) -4-oxobutoxy) amino] acetyl] amino] -2-oxo-1 -Azetidinesulfonic acid and potassium salts (see Example 166) were removed with protecting groups with trifluoroacetic acid and anizole to obtain the target compound with a melting point of 200 캜 or higher.

Example 172

(S) -3-[[2- (aminomethyl) benzoyl] amino] -2-oxo-1-azetidinesulfonic acid, intramolecular salt

(S) -2-oxo-3-[[2-[[[(phenylmethoxy) carbonyl] amino) methyl] benzoyl] amino) -1-azetidinesulfonic acid, potassium salts with hydrogen gas, activated carbon based palladium and The protecting group was removed with hydrochloric acid to obtain a target compound having a melting point of 162 ° to 165 ° C.

Example 173

(S) -3-[[[2- (4-formyl) -1-piperazinyl (-5-hydroxy-pyrido [2,3-d] pyrimidin-6-yl] carbonyl] amino ) -2-oxo-1-azetidinesulfonic acid, potassium salt

(S) -3-amino-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt (see Example 6A) to 2- (4-formyl-1-pipepelinyl) -5-hydroxy-6- [ It was combined with (4-nitrophenoxy) carbonyl] pyrido [2,3-d] pyrimidine and treated with potassium sulfate perfluorobutane in acetone to obtain a target compound having a melting point of 290 ° C. (decomposition).

Example 174

(3S-trans) -α-[[(4-methyl-2-oxo-1-sulfo-3-azetidinyl) amino) carbonyl] benzeneacetic acid, dipotassium salt

(3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (see Example 139) was combined with α- (carboxy) benzene acetyl chloride followed by triethylamine and potassium sulfate Treatment with perfluorobutane gave the target compound at melting point of 147 占 폚 (decomposition).

Example 175

[3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid

A) α- (t-butoxycarbonylamino) -β-cyclohexyl-β-hydroxy-threo-propionic acid

15 g of β-cyclohexyl-α-amino-β-hydroxy-threo-propionic acid was suspended in 150 ml of acetonitrile and 70 ml of water. 17.8 g of triethylamine was added thereto, and the mixture was heated to 60 ° C while stirring. A clear solution was obtained at this temperature, to which 21.0 g of di-t-butylpyrocarbonate was added, followed by continued stirring at 60 ° C. for 1.5 hours. The solvent was removed in vacuo and 50 ml of water was added thereto. The aqueous layer was adjusted to pH 2 by addition of 3N HCl, extracted with ethyl acetate, the organic layer was separated, dehydrated with Na ₂ SO ₄ and evaporated to dryness. Petroleum ether was added to the remaining crystalline material and filtered to obtain 20.4 g of the target compound having a melting point of 113 ° to 115 ° C.

B) α- (t-butoxycarbonylamino) -β-cyclohexyl-β-hydroxy-N-methoxy-threo-tropionamide

20.2 g of α- (t-butoxycarbonylamino) -β-cyclohexyl-β-hydroxy-threo-propionic acid and 7.6 g of O-methylhydroxylamine hydrochloride were suspended in 350 ml of water and 175 ml of t-butanone. Potassium carbonate was added to adjust the pH of the mixture to 4, then 16.4 g of 1-ethyl-3- (3-dimethylaminopropyl) carbodiimide was added and stirred for 1.5 hours while maintaining the pH at 4. T-butanol was removed in vacuo, and the remaining aqueous solution was saturated with sodium chloride and extracted twice with 100 ml of ethyl acetate. The combined organic layers were dehydrated with Na ₂ SO ₄ and then evaporated to dryness. Petroleum ether was added to the remaining crystals, followed by filtration to obtain 18.6 g of the target compound having a melting point of 125 ° to 127 ° C.

C) α- (t-butoxycarbonylamino) -β-cyclohexyl-β- (methanesulfonyloxy) -N-methoxy-threo-propionamide

18.3 g of α- (t-butoxycarbonylamino) -β-cyclohexyl-β-hydroxy-N-methoxy-threo-propionamide was dissolved in 100 ml of anhydrous pyridine with stirring. The solution was cooled to 0 ° C. while stirring, and 9.3 g of methanesulfonyl chloride was added thereto. After holding at 0 ° C. for 1 hour, to this was further added 3.3 g of methanesulfonyl chloride and further stirred for 1 hour. The solution was poured into 300 ml of ice water, 200 ml of ethyl acetate was added, and then diluted sulfuric acid was adjusted to pH 3. The organic layer was separated, dehydrated with Na ₂ SO ₄ and then the solvent was removed in vacuo. The remaining solid was added with petroleum ether and then filtered to obtain 19.0 g of the target compound having a melting point of 150 ° C to 152 ° C.

D) [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-1-methoxy-2-azetadinone

18.7 g of α- (t-butoxycarbonylamino) -β-cyclohexyl-β- (methanesulfonyloxy) -N-methoxy-threo-propionamide is dissolved in 500 ml of anhydrous acetone, and potassium carbonate 9.8 g was added and then the suspension was heated to reflux for 5 hours with stirring. The insoluble inorganic material was filtered off, the solvent was removed in vacuo, and the remaining oily material was dissolved in 30 ml of ethyl acetate. The compound precipitated by addition of petroleum ether was filtered to obtain 12.9 g of the target compound having a melting point of 110 ° to 112 ° C.

E) [3S-trans] -3- (t-botoxycarbonylamino) -4-cyclohexyl-2-azetidinone

1 g of [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-1-methoxy-2-azetidinone is added to 50 ml of ammonia with stirring, and 0.154 g of sodium is added thereto. Divided into ~ 6 portions and added within 5 minutes. Then, 0.025 g more sodium was added and stirring continued for 5 minutes. 0.89 g of ammonium chloride was added thereto, and ammonia was removed. The residue was extracted with warm ethyl acetate, the organic extract was evaporated to dryness, and then petroleum ether was added to the resulting crystals and filtered to give 0.5 g of the target compound having a melting point of 130 ° to 1328.

F) [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid, pyridine salt

5.3 g of [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-2-azetidinone is dissolved in 20 ml of methylene chloride and 80 ml of dimethylformamide, and the solution contains pyridine sulfur trioxide complex 60 After adding mmol, the solution was stirred for 6 hours at room temperature. The solvent was removed in vacuo to give 11.3 g of an oily target compound.

G) [3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid, tetrabutylammonium salt

[3S-trans] -3- (t-butoxycarbonylamino) -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid and 11.3 g of pyridine salt are dissolved in 250 ml of water, and tetrabutyl hydrogen sulfate is added thereto. 9.0 g ammonium was added with stirring, and then the pH was adjusted to 6.5 by addition of 1N potassium hydroxide. The aqueous solution was extracted twice with 200 ml of methylene chloride, the oil layer was dehydrated with Na ₂ SO ₄ , filtered, and the solvent was distilled off to obtain 8 g of the target compound having a melting point of 135 ° C to 138 ° C.

H) [3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid

3.8 g of [3S-trans] -3- (t-butoxycarbonylamino] -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid and tetrabutylammonium salt were stirred in 20 ml of formic acid for 3 hours, and then 20 ml of methylene chloride was added to the mixture, and 1.0 g of the target compound precipitated was filtered out.

Example 176

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) -methoxyimino) acetyl] amino] -4-cyclohexyl-2-oxo-1-azetidine Sulfonic acid, potassium salt

0.25 g of [3S-trans-3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid was dissolved in 30 ml of anhydrous dimethylformamide and 0.12 g of triethylamine. When a clear solution was obtained, 0.2 g of (Z) -2-amino-α- (methoxyimino) -4-thiazole acetic acid, 0.16 g of hydroxybenzotriazole and 0.42 g of dicyclohexylcarbodiimide were added, And stirred for 48 hours at ambient temperature. The precipitated urene was filtered off and the solvent was removed in vacuo. The residue was dissolved in 10 ml of acetone and then 0.41 g of potassium sulfate perfluorobutane was added. After adding 50 ml of ether, the target compound was precipitated, and it was filtered and then column chromatographed with HP-20 using water / acetone (9: 1) as the eluent, and the melting point was 200 ° to 205 ° C. (after freeze drying). 0.36 g of

Example 177

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazole) [(1-carboxy-methylethoxy) imino] acetyl] amino] -4-cyclohexyl- 2-oxo-1-azetidinesulfonic acid, dipotassium salt

A) [3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-diphenylmethoxycarbonyl-4-methylethoxy) imino) acetyl ] Amino] -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid, potassium salt

0.2 g of 3S-trans-3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid (see Example 175) was dissolved in 30 ml of dimethyl formamide and 0.09 g of triethylamine. 0.12 g of hydroxybenzotriazole (Z) -2-amino-α-[(2-diphenylmethoxycarbonyl-1-methylethoxy) imino] -4-thiazole acetic acid 0.30 g and dicyclo 0.33 g of hexylcarbodiimide was added and then stirred at ambient temperature for 12 hours. The precipitated urea is filtered off and the mother liquor is evaporated to dryness. The oily residue was dissolved in 5 ml of acetone, treated with 0.3 g of potassium sulfate perfluorobutane and then poured into 100 ml of ether with stirring. The precipitate was filtered off to give [3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl)-(1-diphenylmethoxycarbonyl-1-methylethoxy) imino ] Acetyl] amino] -4-cyclohexyl-2-oxo-1-azetidinesulfonic acid and potassium salt 0.61g were obtained.

B) [3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) [(1-carboxy-1-methylethoxy) imino] acetyl] amino) -4 -Cyclohexyl-2-oxo-1-azetidinesulfonic acid, dipotassium salt

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) (1-diphenylmethoxycarbonyl-1-methylethoxy) imino] acetyl] amino] 0.61 g of 4-cyclohexyl-2-oxo-1-azetidinesulfonic acid and potassium salt were suspended in 6 ml of anisol, cooled to −15 ° C., and 5 ml of trifluoroacetic acid was added dropwise thereto under stirring. This temperature was maintained for 1 hour and then lowered to -30 ° C. To this, about 100 ml of anhydrous ether was added at a rate such that the temperature did not exceed -10 占 폚. The precipitated compound was filtrated and chromatographed with HP-20 using water / acetone (9: 1) as the eluent to obtain 0.3 g of the target compound (melting point 111 ° to 120 ° C. (decomposition) (after freeze drying). .

Example 178

[3S- [3α, 4β]]-4-cyclohexyl-3-[[[[[3- (furanylmethylene) amino] -2-oxo-1-imidazolidinyl] carbonyl] imino) phenyl Acetyl] amino-2-oxo-1-acetidinesulfonic acid, potassium salt

0.1 g of [3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetidinesulfonic acid (see Example 175) was dissolved in 30 ml of anhydrous dimethylformamide and 0.05 g of triethylamine while stirring. To this solution was added 0.14 g of [[[[(2-furanylmethylene) amino] -2-oxo-1-imidazolidinyl] carbonyl] amino) phenylacetic acid, 0.06 g of hydroxybenzotriazole and dicyclohexyl. 0.17 g of carbodiimide was added and the solution was stirred for 5 days at room temperature. The solvent was removed in vacuo, the residue was dissolved in 10 ml of acetone and the precipitate urea was filtered off. 0.15 g of potassium sulfate perfluoro butane was added to the mother liquor, stirred, and diluted with 50 ml of ether. The precipitate was filtered and chromatographed with HP-20 resin using water / acetone (9: 1) as the eluent (after freeze drying) to obtain 0.14 g of a product having a melting point of 195 ° to 200 ° C. (decomposition).

Example 179

[3S- [3α (R ^* ), 4β]]-4-cyclohexyl-3-[[3- (4-ethyl-2,3-dioxo-1-piperazinyl) -1,3-dioxo 2-phenylpropyl] amino) -2-oxo-1-azetidinesulfonic acid, potassium salt

0.1 g of [3S-trans] -3-amino-4-cyclohexyl-2-oxo-1-azetinylsulfonic acid (see Example 175) was dissolved in 30 ml of dimethylformamide and 0.5 g of triethylamine with stirring. (R) -α-[[4-ethyl-2,3-dioxo-1-piperazinyl) carbonyl] aminobenzeneacetic acid, 0.06 g of hydroxybenzotriazole and 0.17 g of dicyclohexylcarbodiimide are added And stirred at room temperature for about 16 hours. The solvent was distilled off in vacuo and the oily residue was dissolved in 10 ml of acetone. The precipitated urea was filtered off, 0.15 g of potassium sulfate perfluorobutane was added to the mother liquor, stirred, and diluted with 50 ml of ether. The precipitate was filtered, chromatographed with HP-20 resin using water / acetone (9: 1) as the eluent, and lyophilized to obtain 0.15 g of the target compound having a melting point of 175 ° C to 180 ° C.

Example 180

(Trans, Z) -3-[[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-ethyl-2-oxo-1-acetidinesulfonic acid, potassium salt

(Z) -2-amino-α- (meth instead of (Z) -2-amino-α-[(1-diphenylmethoxycarbonyl-1-methylethoxy) imino] -4-diazolacetic acid It carried out according to the method of Example 170 using the oxyimino) -4-thiazole acetic acid, and obtained the target compound of melting | fusing point 190 degreeC (decomposition).

Example 181

(±)-(trans) -3-amino-2-oxo-4-phenyl-1-azetidinesulfonic acid

A) (±)-(trans) -2-oxo-4-phenyl-1-azetidine-t-butyldiphenylsilane

A solution of 20.56 g of t-butylchlorodiphenylsilane in 45 ml of dimethylformamide was cooled to 0 ° C. in the presence of argon, followed by 10.4 ml of triethylamine, followed by (±) -2-oxo-4-phenyl-1 -Azetidine was added. After holding at 0 ° C. for several hours, 1 ml of triethylamine and 2.11 g of t-butylchlorodiphenylsilane were added to the resulting mixture and stirred at 5 ° C. for 65 hours. The reaction mixture was poured into 300 ml of ice water, and extracted three times with 125 ml of ether-ethyl acetate (3: 1). The organic extract was washed three times with 50 ml of phosphate buffer (pH4.5), once with 50 ml of saturated sodium bicarbonate solution, twice with 50 ml of water and with saturated sodium chloride solution, and then dehydrated with Na ₂ SO ₄ . After filtration, the solid obtained by concentration in vacuo was washed with hexane and dried in high vacuum to obtain 15 g of the target compound in the solid phase.

B) (±)-(trans) -3-azido-2-oxo-4-phenyl-1-azetidine-t-butyldiphenylsilane

A 500 ml flask equipped with a stirring rod, a gas inlet and a diaphragm was dried with a flame in the presence of argon, and then 0.65 ml of a 1.6 Mn-butyllithium solution dissolved in hexane was added thereto, cooled to -40 ° C, and then, tetra It was dissolved in 2 ml of hydrofuran. To this was added 0.16 ml of diisopropylamine dropwise and the resulting mixture was stirred for 30 minutes and then cooled to -78 ° C. To this was added dropwise a solution of 400 mg of (±)-(trans) -2-oxo-4-phenyl-1-azetidine-t-butyldiphenylsilane in 1.5 ml of tetrahydrofran over about 5 minutes. After further stirring for 20 minutes, the solution was treated with 204 mg of p-toluenesulfonylazide dissolved in 0.5 ml of tetrahydrofuran. The resulting mixture was stirred at −78 ° C. for 10 minutes and then 0.4 ml of chlorotrimethylsilane was added dropwise. After further stirring for 10 minutes, the cooling bath was removed and the reaction mixture was stirred at ambient temperature for 2.5 hours. 20 ml of ethyl acetate, and then 8 ml of phosphate buffer (pH4.5) were added to this mixture, cooling at 0 degreeC. The organic layer was washed twice with 8 ml of phosphate buffer, three times with 10 ml of 5% sodium bicarbonate solution, once with 10 ml of 50% sodium chloride solution, and once with 10 ml of saturated sodium chloride solution, followed by dehydration with Na ₂ SO ₄ . After filtration, 500 g of the oily substance obtained by concentration in vacuo was flash chromatographed with 5% ethyl acetate-hexane to obtain 253 mg of the target compound.

C) (±)-(trans) -3-azido-2-oxo-4-phenyl-1-azetidine

17 g of crude (±)-(trans) -3-azido-2-oxo-4-phenyl-1-azetidine-t-butyldiphenylsilane was dissolved in 240 ml of methanol, and 35 ml of concentrated hydrochloric acid was added dropwise at 0 ° C. did. The cooling bath was removed and the reaction stirred at ambient temperature for 1 hour, then cooled to 0 ° C. and saturated sodium bicarbonate solution was added until neutral. The mixture was extracted once with 300 ml of ethyl acetate and four times with 10 ml each. The organic extract was washed with 5% sodium bicarbonate, 50% sodium chloride (1: 1) solution, saturated sodium chloride solution and dehydrated with Na ₂ SO ₄ . . The mixture was filtered, and 15 g of heavy oil obtained by concentration in vacuo was chromatographed with 100 g of silica gel using 20% ethyl acetate-hexane to give 460 mg of the target compound.

D) (±)-(trans) -3-azido-4-phenyl-1-azetidinesulfonic acid, tetrabutylammonium salt

A solution of 300 mg of (±)-(trans) -3-azido-2-oxo-4-phenyl-1-azetidine in 3 ml of dimethylformamide was cooled to 0 ° C. in the presence of argon, and dimethylformamide was added thereto. And 4.78 ml of a complex of sulfur trioxide (0.5 M solution dissolved in dimethylformamide) was added dropwise. The cold bath was removed and the reaction mixture was stirred at ambient temperature for 2 hours and then poured into 80 ml of 0.5M potassium phosphate monobasic (pH5.5). The solution was extracted with dichloromethane, followed by tilting the dichloromethane layer and 514 mg of tetrabutylammonium disulfide was added. The mixture was extracted with dichloromethane and the organic extracts were washed with 10% sodium chloride solution and then dehydrated with Na ₂ SO ₄ . It was filtered and then concentrated in vacuo to yield 800 mg of oil, of which about 40% was the target compound and the remainder was dimethylformamide. This mixture was used in the next step without purification.

E) (±)-(trans) -3-amino-4-phenyl-1-azetidinesulfonic acid

A solution obtained by dissolving (±)-(trans) -3-amino-4-phenyl-1-azetidinesulfonic acid and tetrabutylammonium salt in 4 ml of methanol was hydrogenated in presence of 30 mg of platinum oxide at 1 atmosphere and room temperature. After 15 minutes the system was degassed and then fresh hydrogen was introduced. After 45 minutes the reaction was complete and the reaction system was filled with nitrogen. After several days in 200 ml of dichloromethane-methanol (4: 1) at room temperature, the catalyst was filtered off when it was completely coagulated. The filtrate was concentrated in vacuo until 18 ml and 0.2 ml of 97% formic acid was added thereto.

After cooling at 5 ° C. for several hours, the resulting solid was filtered, washed with dichloromethane and dried to give 150 g of the desired compound in the solid phase.

Analysis of C ₉ H ₁₉ N ₂ O ₄ S

Theoretic value: C 44.62; H 4.17; Found: C 43.36; H 4.31;

N 11.57; S 13.23 H 11.09; S 13.02

Example 182

(±)-(trans) -2-oxo-4-phenyl-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid potassium salt

A) (±)-(trans) -2-oxo-4-phenyl-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid tetrabutylammonium salt

A solution of 52 mg of N-hydroxybenzotriazole monohydrate and 46 mg of phenylacetic acid in 0.3 ml of dimethylformamide was treated with 70 mg of a dicyclohexylcarbodiimide solid at 0 ° C. in the presence of nitrogen. A cold bath was taken and the mixture was stirred at ambient temperature for 1 hour. Dilute this mixture with 0.3 ml of dimethylformamide, and then add 75 mg of (±)-(trans) -3amino-2-oxo-4-phenyl-1-azetidinesulfonic acid (see Example 181) to solid form. Then, 0.05 ml of triethylamine was added dropwise. The reaction was stirred at ambient temperature for 23 hours, then filtered, and the filter cake was washed with dimethylformamide. The filtrate is added to 20 ml of 0.5 M monobasic potassium phosphate (pH 4.5), and the mixture is washed three times with 8 ml of ethyl acetate, followed by an ethyl acetate layer, followed by tetrabutylammonium bisulfate (0.31 mmol). ) 105 mg was added. The mixture was extracted three times with 15 ml of dichloromethane, and the extract was washed twice with 15 ml of 10% sodium chloride solution, once with 10 ml of saturated sodium chloride solution, and then dehydrated with Na ₂ SO ₄ . It was filtered, concentrated in vacuo and heated at high vacuum at 32 ° C. to yield 165 mg of oil, of which about 40% was the target compound and 60% was dimethylformamide.

B) (±)-(trans) -2-oxo-4-phenyl-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

41 mg of potassium perfluorobutane sulfate in a solution of (±) -trans-2-oxo-4-phenyl-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid and tetrabutylammonium salt dissolved in 1.5 ml of acetone ( 0.121 mmol) was added, and the glassy material obtained by dilution with 12 ml of ether was titrated with ether to obtain 43 mg of a solid, and about 20% of the solid was an impurity containing a tetrabutylammonium component. This solid was dissolved in 50% acetone aqueous solution and passed through 1 ml of Dowex 50W-X2K ⁺ ion exchange resin. The solid obtained by removing the solvent was washed with acetone and hexane and dried in a high vacuum at 60 ° C. to obtain 15 mg of the target compound.

Analysis of C ₁₇ H ₁₅ N ₂ O ₅ SK

Theoretic value: C 31.23; H 3.80 Found: C 50.44; H 4.20

N 7.03; S 8.05; N 7.01; S 7.59

K 9.81 K 9.40

Example 183

(±)-(trans, Z) -3-[[2-amino-4-thiazolyl) (methoxyimino) acetyl] amino) -2-oxo-4-phenyl-1-azetidinesulfonic acid, potassium salt

In a solution of N-hydroxybenzotriazole, 52 mg of hydrate and 69 mg of (Z) -2-amino-α- (methoxyimino) -4-thiazole acetic acid dissolved in 0.3 ml of dimethylformamide at ambient temperature in the presence of argon 70 mg of dicyclohexylcarbodiimide solid was added. The mixture was stirred for 1 hour and 75 mg of (±)-(trans) -3-amino-2-oxo-4-phenyl-1-azetidinesulfonic acid (see Example 181) was added in the form of a solid, 0.05 ml of triethylamine was added dropwise. The reaction was stirred at ambient temperature for 23 hours, then dimethylformamide was removed at 30 ° C. in high vacuum, the residue was titrated with 2 ml of acetone and filtered. The filter cake was washed twice with 3 ml of acetone, and 86 mg of potassium sulfate perfluorobutane was added to the filtrate. The rubbery solid obtained by diluting with 10 ml of ether was titrated with acetone and hexane, then washed and dried to give 82 mg of the target compound as a solid.

Analysis of C ₁₅ H ₁₄ N ₅ O ₆ SK

Theoretic value: C 40.26; H 3.16 Found: C 38.60; H 3.19

N 15.65; S 14.33 N 15.07; S 13.87

K 8.74 K 7.5

Example 184

(Cis) -2-oxo-4-phenyl-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

A) N-benzylidene-2,4-dimethoxybenzylamine

12.0 g of 2,4-dimethoxybenzylamine hydrochloride was added to 100 ml of 1N sodium hydroxide solution, and the mixture was extracted with 125 ml of ethyl acetate. The organic layer was dehydrated with anhydrous sodium sulfate, and the solvent was evaporated to remove 10.2 g of oily 2,4-dimethoxybenzylamine. This amine was dissolved in 150 ml of benzene, and 6.47 g of benzaldehyde and 0.6 g of p-toluenesulfonic acid monohydrate were added thereto. The mixture was heated to reflux to remove water with a Dean-Stark separator, and within 1.1 hours the amount of 1.1 ml of water was separated. The mixture was cooled at room temperature and the benzene solution was further cooled to give some precipitate. Benzene was removed under reduced pressure and 60 ml of petroleum ether was added to the residue. The oil layer was further precipitated to separate, and 10 ml of benzene was added to make the layer homogeneous and the remaining precipitate was filtered off. The solvent was evaporated from the filtrate to obtain 14.2 g of an oily target compound.

B) (±)-(cis) -4-phenyl-1- (2,4-dimethoxybenzyl) -2-oxo-3-azidoazetidine

1.02 g (4.0 mmol) of imine N-benzylidene-2,4-dimethoxybenzylamine in which 1.62 g of azidoacetic acid was dissolved in 25 ml of methylene chloride in the presence of nitrogen and dissolved in 3.24 g of triethylamine and 10 ml of methylene chloride in this solution. ) Was added. The mixture was cooled in an ice bath, and slowly added 3.36 g of trifluoroacetic anhydride to it, and the solution turned dark color. The mixture was stirred for 1 hour in an ice bath, then allowed to warm to room temperature and further stirred for 15 minutes. The solution was washed twice with 50 ml of 5% NaHCO ₃ solution once with 60 ml of water followed by 60 ml of 1N HCl solution. The organic layer was dehydrated with anhydrous sodium sulfate, and the solvent was evaporated to remove 1.72 g of a dark colored rubbery crude product. This rubbery material was treated several times with activated carbon, and the resulting navy blue mixture was eluted with 40 g of silica gel with petroleum ether: ethyl acetate (1: 1) to perform chromatography. Fractions were combined and quenched in anhydrous ice-acetone bath to form crystals. This crystal was used as a seed and recrystallized from petroleum ether-ethyl acetate to obtain 817 mg of the target compound in the form of a needle, which was dissolved when warmed to room temperature.

C) (±)-(cis) -4-phenyl-2-oxo-3-azidoazetidine

737 mg of (±)-(cis) -4-phenyl-1- (2,4-dimethoxybenzyl) -2-oxo-3-azidoazetidine was dissolved in 25 ml of acetonitrile and then 80 ° C. in the presence of nitrogen. Heated to 83 ° C. To this solution, 943 mg of potassium persulfate and 570 mg of potassium monohydrogen phosphate dissolved in 25 ml of water were added over 1 hour.

The mixture was cooled and the pH was adjusted to 6-7 by the addition of solid potassium monohydrogen phosphate, then most of the acetonitrile was removed under reduced pressure, and the resulting mixture was extracted with 60 m of chloroform. The chloroform layer was washed with 60 ml of water, dehydrated with anhydrous sodium sulfate, and then the solvent was evaporated off to obtain an oily crude product.

Chromatography was performed by eluting with 40 g of silica gel with petroleum ether-ethyl acetate (1: 1), and the crystals obtained from the fractions were recrystallized with petroleum ether-ethyl acetate to obtain 267 mg of the target compound.

D) (±)-(cis) -4-phenyl-2-oxo-3-azido-1-azetidinesulfonic acid, tetrabutylammonium salt

162 mg of (±)-(cis) -4-phenyl-2-oxo-3-azidoazetidine was cooled to 0 ° C. under a nitrogen atmosphere and dissolved in dimethylformamide therein, about 0.5 M dimethylformamide-sulfuric acid 3.5 ml of the complex solution was added dropwise by syringe. The resulting clear solution was stirred at 0 ° C. for 15 minutes, then the mixture was poured into 50 ml of 0.5 M monobasic potassium phosphate solution and washed three times with 50 ml of methylene chloride. 292 mg of tetra-n-butylammonium disulfate was added to this aqueous solution, and the mixture was extracted 6 times with 50 ml of methylene chloride. The methylene chloride layers were combined, dehydrated with anhydrous sodium sulfate, and the solvent was evaporated to remove 272 mg of rubbery target compound.

E) (±)-(cis) -2-oxo-4-phenyl-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid, potassium salt

293 mg of (±)-(cis) -4-phenyl-2-oxo-azido-1-azetidinesulfonic acid and tetrabutylammonium were dissolved in 4 ml of ethanol, and hydrogenated with 80 mg of a platinum oxide catalyst at 1 atmosphere. After stirring for 1 hour, the catalyst was filtered through a Celite-treated Millipore filter where some catalyst particles passed through the filter and the filtrate became black. Ethanol was removed under reduced pressure, and the residue was dissolved in 4 ml of dimethylformamide. 81 mg of N-hydroxy benzotriazole monohydrate, 78 mg of phenylacetic acid and 117 mg of dicyclohexylcarbodiimide were added thereto, and the mixture was stirred under a nitrogen atmosphere for about 16 hours. The slurry was evaporated in vacuo and then the slurry obtained by titration with 10 ml of acetone was filtered through a Millifor filter treated with celite, and the brown filtrate obtained was treated with 193 mg of potassium sulfate perfluorobutane. After 20 ml of ether was added thereto, the rubbery material was separated. The liquid was removed and the gummy material was washed with ether. A gummy substance was dissolved in 10 ml of methanol and a small amount of precipitate formed after the addition of ether. The mixture was filtered and the colored filtrate was further treated with ether. The formed precipitate was filtered and recrystallized twice with ether-methanol to give 26 mg of the target compound.

Analysis of C ₁₇ H ₁₅ O ₅ N ₂ SK · 2H ₂ · O

Theoretic value: C 46.99; H 4.41; Found: C 47.24; H 4.19;

N 6.45 N 6.34

Example 185

(Cis, Z) -3-[[(2-amino-4-thiazolyl) (methoxy imino) acetyl] amino] -2-oxo-4-phenyl-1-azetidinesulfonic acid, potassium salt

Dissolve 560 mg of (cis) -2-oxo-4-phenyl-3-[(phenylacetyl) amino] -1-azetidinesulfonic acid and potassium salt (see Example 184, Part D) in 5 ml of ethanol and at 1 atm. Hydrogenated with 110 mg of platinum oxide catalyst. After stirring for 1 hour, the catalyst was filtered through a Millipore filter treated with celite, and the catalyst particles passed through the filter and the filtrate became black. Ethanol was removed under reduced pressure and the residue was dissolved in 4 ml of dimethylformamide. To this, 168 mg of N-hydroxybenzotriazole monohydrate, 221 mg of (Z) -2-amino-α- (methoxyimino) -4-thiazole acetic acid and 227 mg of dicyclohexylcarbodiimide were added and nitrogen was present. Under stirring for about 16 hours. The slurry was evaporated in vacuo and the slurry obtained by titration with 15 ml of acetone was filtered through a Millipore filter treated with Celite, and then 372 mg of potassium sulfate perfluorobutane was added to the filtrate. To this was added 15 ml of ether, after which the gummy material was separated out. The liquid was removed and the gummy material was washed with ether. The rubbery material was dissolved in 5 ml of water and eluted with water using 150 ml of HP-20 resin. The fractions 16-34 (each 30 ml each) were combined and lyophilized to obtain 201 mg of the target compound as a solid.

H₂O에 대한 분석C ₁₅ H ₁₄ O ₆ N ₅ S ₂ K

Assay for H ₂ O

Theoretical value: C 36.73; H 3.49; Found: C 36.65; H 3.00;

N 14.28; S 13.07 N 13.99; S 13.48

K 7.97 K 8.30

Example 186

(Cis) -3-amino-2-oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid

A) N- (3-phenyl-2-propenylidene) -4-methoxyaniline

12.32 g of p-anisidine was dissolved in 160 ml of methylene chloride, and 20 g of anhydrous magnesium sulfate was added thereto. The mixture was cooled in an ice bath and then 13.22 g of trans-cinnaaldehyde was added. The mixture was stirred in the presence of nitrogen for 2 hours, filtered and the filtrate was evaporated to give a solid. This crude product was recrystallized from methylene chloride-petroleum ether to give 20.96 g of the target compound in the solid phase.

B) (±)-(cis) -3-azido-1- (4-methoxyphenyl) -2-oxo-4- (2-phenylethenyl) azetidine

24.26 g of 2-azido acetate was dissolved in 100 ml of methylene chloride and the solution was cooled in an ice-ice bath. 48.57 g of triethylamine dissolved in 250 ml of methylene chloride and 14.24 g of N- (3-phenyl-2-propenylidene) -4-methoxyaniline were added to the solution. 50.41 g of trifluoro acetic anhydride was added dropwise to the solution over 1 hour, stirred in an ice bath for 1 hour, then warmed to room temperature and stirred for about 16 hours. The mixture was diluted with 250 ml of methylene chloride, washed once with 750 ml of water, twice with 750 ml of 5% sodium bicarbonate solution, and with 750 ml of 1N HCl solution. The organic layer was dehydrated with anhydrous sodium sulfate, and then the crude product (solid) obtained by evaporating off the solvent was recrystallized with ethyl acetate to obtain 111.39 g of the target compound in the solid phase.

C) (±)-(cis) -3-azido-2-oxo-4- (2-phenylethenyl) azetidine

(±)-(cis) -3-azido-1- (4-methoxyphenyl) -2-oxo-4 dissolved in 65 ml of acetonitrile at 0 ° C. in a solution of 10.22 g of ceric ammonium nitrate dissolved in 13 ml of water. 1.99 g of-(2-phenylethenyl) azetidine was added for 15 minutes. 10 ml of acetonitrile was further added and used for washing. The mixture was further stirred at 0 ° C. for 15 minutes, diluted with 750 ml of ethyl acetate, washed six times with 600 ml of water, dehydrated with anhydrous sodium sulfate, and the solvent was evaporated off to obtain an oily substance. The crude product was chromatographed with 90 g of silica gel and eluted first with 250 ml of 30% ethyl acetate / petroleum ether and then with 50% ethyl acetate / petroleum ether. Fractions 11-16 (50 ml each) were combined and evaporated to afford 802 mg of the target compound as an oil.

D) (±)-(cis) -3-azido-2-oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid, tetra-n-butylammonium salt

334 mg of (±)-(cis) -3-azido-2-oxo-4- (2-phenylethenyl) azetidine was dissolved in 3 ml of dimethylformamide, and 868 mg of pyridine-sulfur trioxide complex was added thereto. The mixture was stirred at room temperature in the presence of nitrogen, then poured into 200 ml of 0.5 M monobasic potassium phosphate solution and washed with 30 ml of methylene chloride. 530 mg of tetra-n-butyl ammonium bisulfate was added to the aqueous solution, and the mixture was extracted four times with 50 ml of methylene chloride. The organic layers were combined, backwashed twice with 100 ml of water, dehydrated with anhydrous magnesium sulfate, and the solvent was evaporated to remove 824 mg of the gummy target compound.

E) (±)-(cis) -3-azido-2-oxo-4- (2-phenylethenyl-1-azetidinesulfonic acid

300 mg of (±)-(cis) -3-azido-2-oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid and tetra-n-butylammonium salt were dissolved in 4 ml of tetrahydrofuran and rapidly Was stirred. To this mixture was added 600 mg of zinc dust, followed by 0.8 ml of 1N monobasic potassium phosphate solution, which was heated to 45 ° C. and then stirred for 3 hours at ionicity. The mixture was filtered, the filtrate was dissolved in 40 ml of methylene chloride and 10 ml of water, the aqueous layer was extracted three times with 40 ml of methylene chloride, and the methylene chloride layers were combined and the solvent was evaporated off to give 256 mg of foamy material. The crude water was dissolved in a small amount of about 30% acetone / water and eluted with 40 ml of water using 7.5 ml of Dowex (K ⁺ ) resin (0.7 meq./vnl). The eluate was evaporated to dissolve 151 mg of foamy material in 2 ml of water, and then adjusted to pH 2 by addition of 1N HCl solution. A small amount of acetonitrile was added thereto to dissolve the precipitate, and the solution was eluted with 150 ml of water followed by 10% acetone / water using 15 ml of HP-20 resin. Fractions 2 to 13 (15 ml each) were combined and evaporated to yield 101 mg of the target compound in the foam.

Example 187

(±)-(cis, Z) -3-[[2-amino-4-thiazolyl) (methoxyimino) -acetyl] amino] -2-oxo-4- (2-phenylethenyl) -1- Azetidinesulfonic acid, potassium salt

Dicyclohexylcarbodiimide 69 mg in a solution of 68 mg of (Z) -2-amino-α- (methoxyimino) -4-thiazole acetic acid and 51 mg of N-hydroxybenzotriazole monohydrate dissolved in 2 ml of dimethylformamide. Was added. The mixture was stirred for 30 minutes at room temperature in the presence of nitrogen, followed by (cis) -3-amino-2-oxo-4- (2-phenylethenyl) -1-azetidinesulfonic acid (see Example 186). 90 mg and 34 mg triethylamine were added and stirred for 20 hours in the presence of nitrogen. This slurry was evaporated in vacuo, titrated with 10 ml of acetone, filtered and 113 mg of potassium sulfate perfluorobutane was added to the filtrate. When diluted with 30 ml of ether, 169 mg of the solid product obtained by filtration was dissolved in a small amount of about 10% acetonitrile / water, and then eluted with 150 ml of water followed by 10% acetone / water using 34 ml of HP-20 resin. Fractions 16 to 19 (15 ml each) were combined and the solvent was evaporated to remove 110 mg of the target compound as a solid.

Analysis of C ₁₇ H ₁₆ O ₆ N ₆ S ₂ KH ₂ KH ₂ O

Theoretic value: C 40.23; H 3.57; Found: C 40.03; H 3.05;

N 13.80; S 12.63; N 13.61; S 12.31;

K 7.70 K 7.56

Example 188

(Cis) -3-amino-4- (methoxycarbonyl) -2-oxo-1-azetidinesulfonic acid

A) [(4-methoxyphenyl) amino] acetic acid, methyl ester

A 1 L three-necked flask equipped with a nitrogen inlet and a stirring rod was charged with 56.88 g of MgSO ₄ , followed by a solution of 19.43 g of recrystallized anizidine dissolved in 250 ml of dichloromethane. After cooling to 0 ° C., 19.92 g of methylglyoxalate hemiacetam dissolved in 250 ml of dichloromethane was added over 1.5 hours, further stirred at 0 ° C. for 20 minutes, and then the reaction mixture was suction filtered, and sodium sulfate Dehydrated, filtered and concentrated in vacuo to 1/4 volume. 300 ml of nucleic acid was added thereto, and the oily substance obtained by concentrating the solution was allowed to react by being allowed to stand at 5 deg.

B) (cis) -3- (1,3-dihydro-1,3-dioxo-2H-isoindol-2-yl) -4-methoxycarbonyl-2-oxo-1- (4-meth Oxyphenyl) azetidine

Into a dried 500 ml three-necked flask equipped with a stirring rod, an addition funnel, a diaphragm and a nitrogen inlet was filled with a solution of [(4-methoxyphenyl) imino] acetic acid and 21.09 g of methyl ester in 150 ml of dichloromethane. Cool to 0 ° C. To this was added 19.2 ml (0.14 mol) of triethylamine dropwise, and then a solution of 28.4 g of (N-phthalimide) acetyl acid chloride dissolved in 150 ml of dichloromethane was added dropwise over 1 hour. The reaction mixture was stirred at 0 ° C. for 1.5 h and then diluted with 2.5 L of dichloromethane. The organic solution was washed twice with 500 ml of monobasic potassium phosphate (pH 4.5), twice with 500 ml of 5% sodium bicarbonate, and once with 500 ml of saturated sodium chloride solution, and then dehydrated with sodium sulfate. This was filtered and concentrated in vacuo, and the resulting solid was washed with ethyl acetate, cold acetone and nucleic acid to give 18.65 g of product.

C) (cis) -4-methoxycarbonyl) -1- (4-methoxyphenyl) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] azetidine

In a 500 ml dry flask equipped with a nitrogen inlet, a stirring rod and a diaphragm was (cis) -3- (1,3-dioxo-2H-isoindol-2-yl) -4-methoxycarbonyl-2-oxo- 18.65 g of 1- (4-methoxyphenyl) azetidine and 325 ml of dichloromethane were charged. The resulting suspension was cooled to −30 ° C. and then 3.52 ml of methyl hydrazine was added dropwise. The reaction mixture was warmed to 0 ° C. and then stirred for 1 hour. To this was further added 0.4 ml of methyl hydrazine and the mixture was stirred for 10 minutes. This process was repeated several times to add 7.7 ml (2.9 equiv) of all methyl hydrazine. The solvent was removed in vacuo, then 200 ml of dichloromethane were added and the mixture was concentrated again. This process was repeated two more times, and the resulting foamy material was dried in high vacuum for 20 minutes, then dissolved in 225 ml of dichloromethane again and left at ambient temperature for about 16 hours. At this time, a considerable amount of solids precipitated. The mixture was filtered in the presence of nitrogen, the filtrate was cooled to 0 ° C. in the presence of nitrogen, and then 17 ml of diisophthophyllethylamine was added thereto, followed by dropwise addition of 7 ml of benzyl chloroformate. The reaction was stirred at 0 ° C. for 30 minutes and then at ambient temperature for 1.5 hours. The mixture was washed twice with 300 ml of monobasic potassium phosphate buffer (pH 4.5), twice with 300 ml of 5% sodium bicarbonate, and once with 300 ml of saturated potassium chloride solution, then dehydrated with sodium sulfate and filtered. Concentration in vacuo gave the resulting foamy material titrated with ether to give 9.9 g of the desired compound in the solid phase.

D) (cis) -4- (methoxycarbonyl (-2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidine

A solution of 8.59 g of ceric ammonium nitrate dissolved in 60 ml of acetonitrile-water (1: 1) was dissolved in (cis) -4- (methonoxycarbonyl) -1- (4-methoxyphenyl) -2- in 50 ml of acetonitrile. The mixture was treated with a slurry of 2 g of oxo-3-[[(phenylmethoxy) carbonyl] amino] azetidine over 10 minutes. The reaction mixture was further stirred at ambient temperature for 10 minutes and then diluted with 100 ml of ethyl acetate. The separated aqueous layer was washed three times with 40 ml of ethyl acetate, and the combined organic extracts were washed three times with 70 ml of 50% sodium bicarbonate. The basic washing solution was backwashed with 50 ml of ethyl acetate, and the combined organic extracts were washed once with aqueous sodium sulfite solution, twice with 100 ml of 5% sodium carbonate solution, twice with 100 ml of 5% sodium chloride solution, and twice with 50 ml of saturated sodium chloride solution. Darco G-60 activated carbon was added and stirred for 30 minutes. Sodium sulfate was added and the mixture was filtered and concentrated in vacuo, and the oily substance obtained was titrated with ether to give 685 mg of the solid target compound.

E) (cis) -4- (methoxycarbonyl) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidansulfonic acid, tetrabutylammonium salt

100 mg of (cis) -4- (methoxycarbonyl) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidine and 172 mg of pyridine-sulfur trioxide complex are dissolved in 1 ml of pyridine, Stir at 80 ° C. for 3 hours in the presence of argon. The reaction mixture was poured into 70 ml of 0.5 M monobasic potassium phosphate (pH 5.5) and extracted four times with 30 ml of dichloromethane. 122 mg of hydrogen tetrabutylammonium sulfate was added to the aqueous layer, and then extracted four times with 30 ml of dichloromethane. The organic extract was washed with 8% sodium chloride solution, then dehydrated with sodium sulfate and filtered. Concentration in vacuo gave 186 mg of the target compound of the pickled oil.

F) (cis) -3-amino-4- (methoxycarbonyl) -2-oxo-1-azetidinesulfonic acid

A solution of 186 mg of (cis) -4- (methoxycarbonyl) -2-oxo-3-[[(phenylmethoxy) carbonyl] amino] -1-azetidine sulfonic acid and tetrabutylammonium salt dissolved in 2 ml of methanol was prepared. Hydrogenation was carried out for 1.5 hours in the presence of 95 mg of activated carbon based 10% palladium at 1 atm. The catalyst was filtered off and washed with dichloromethane, then the filtrate was treated with 97% formic acid and then cooled to -50 ° C (in this step, the presence of seed crystals to induce crystallization is required). When crystallization started, the mixture was stirred at 10 ° C. for about 16 hours. The resulting solid was washed with dichloromethane, hexanes and dried in vacuo to afford 50 mg of the target compound.

Example 189

(Cis) -3-[[2-amino-4-thiazolyl) [[1- (diphenyl methoxycarbonyl) -1-methylethoxy] imino] acetyl] amino] -4- (methoxycarb Bonyl) -2-oxo-1-azetidinesulfonic acid, potassium salt

34 mg of N-hydroxybenzotriazole hydrate and 101 mg of 2-amino-α-[[1- (diphenylmethoxycarbonyl) -1-methylethoxy imino] -4-thiazole acetic acid 0.5 ml of dimethylformamide The solution dissolved in was treated with 45 mg of dicyclohexylcarbodiimide (solid), and then the mixture was stirred for 45 minutes at ambient temperature in the presence of argon. To this was added 45 mg of (cis) -3-amino-4-methoxycarbonyl) -2-oxo-1-azetidinesulfonic acid (see Example 188) in the form of a solid, followed by dropwise addition of 0.03 ml of triethylamine. did. The reaction was stirred at ambient temperature for about 16 hours, then dimethylformamide was removed at high vacuum and 30 ° C. and the residue was titrated with acetone. 67 mg of potassium sulfate perfluorobutane was added to the supernatant, and the solid obtained by dilution with ether was washed with ether, and then dried in vacuo to obtain 93 mg of the target compound.

Example 190

(Cis) -3-[[(2-amino-4-thiazoline) [(1-carboxy-1-methylethoxy) imino] acetyl] amino] -4-) methoxy carbonyl) -2-oxo -1-azetidinesulfonic acid, dipotassium salt

(Cis) -3-[[2-amino-4-thiazolyl)-[[1- (diphenyl methoxycarbonyl) -1-methylethoxy) imino] acetyl] amino] -4- (methoxy A slurry made by adding carbonyl) -2-oxo-1-azetidinesulfonic acid and potassium salt to 0.4 ml of anisol was stirred at -12 ° C in the presence of argon and then cooled (-10 ° C) trifluoro 0.9 ml of acetic acid was added. After 1.5 h, 4 ml of ether and 2 ml of hexane were added thereto and the slurry was stirred at -10 [deg.] C. for 15 minutes and at ambient temperature for 15 minutes. The resulting solid was isolated by centrifugation and washed with ether. The pH of the suspension suspended in 0.5 ml of cold water was adjusted to 6 by the addition of 1 N potassium hydroxide, eluted with water using a 30 ml HP-20AG column, acetonitrile was added twice, and then evaporated to the desired concentration. 30 mg of compound was obtained.

Analysis of C ₁₄ H ₁₅ K ₂ N ₅ O ₉ S ₂

Theoretic value: C 31.15; H 2.81; Found: C 29.08; H 3.03;

N 12.98 N 12.19

Example 191

(S)-(trans) -3-amino-4-ethynyl-2-oxo-1-azetidinesulfonic acid

A) 2- (trimethylsilyl) ethynyl magnesium bromide

To a 50 ml flask dried with flame under nitrogen constant pressure, 20 ml of anhydrous tetrahydrofuran, 2.20 ml of trimethylsilyl acetylene and 0.05 ml of a 3.06 M methylmagnesium bromide solution dissolved in ether were added. This mixture was stirred for 140 minutes to afford the desired compound.

B) (S)-(trans) -4- [2- (trimethylsilyl) ethynyl] -2-oxo-3-[(triphenylmethyl) amino] azetidine

To a 250 ml three-neck flask dried with flame was added 6.00 g of (S)-(cis) -4- (methylsulfonyl) -2-oxo-3-[(triphenylmethyl) amino] azetidine, The flask was filled with nitrogen and kept at nitrogen constant pressure. The reaction mixture was added dropwise by syringe with vigorous stirring of 4.65 ml of 3.06 M methylmagnesium bromide solution in which anhydrous ice isopropane was cooled in a bath and then dissolved in ether. The 2 = (trimethylsilyl) ethynyl magnesium bromide solution prepared in part A was added to a Teflon tube under nitrogen constant pressure (the flask containing the reaction was washed with 7 ml of tetrahydrofuran). After the addition, the cold bath was removed. After 45 minutes, a solution of 3.5 g of potassium disulfide dissolved in 20 ml of water was added thereto. Most of the tetrahydrofuran was removed in a rotary evaporator. The residue was placed in a separating funnel and partitioned between ether and water. The aqueous layer was separated and then extracted twice with ether. The combined ether layers were washed once with saturated sodium chloride solution, dehydrated with sodium sulfate and filtered. The foamed material obtained by removing the solvent was chromatographed by silica column. Eluted with 2 l of dichloromethane, 1 l of 1% ether / dichloromethane, 2 l of 2% ether / dichloromethane and 1.5 l of 10% ether / dichloromethane (fraction 1 = 1,000 ml, fraction 2,3 = 500 ml, fraction 4-final) = 250 ml), 1.30 g of the target compound in Fraction Zones 2-8, and 1.80 g of the corresponding trans-isomer in Fractions 12-19 were obtained. Fractions 9-11 contain 1.19 g of a mixture of cis and trans isomers.

C) (S)-(trans) -4-ethynyl-2-oxo-3-[(triphenylmethyl) amino] azetidine

2.97 g of (S)-(trans) -4- [2- (trimethylsilyl) ethynyl] -2-oxo-3-[(triphenylmethyl) amino] azetidine is dissolved in 30 ml of dichloromethane and fluorinated therein 330 mg of tetrabutylammonium (containing 20-25% of water) was added. After 20 minutes the solvent was removed in vacuo and the residue was dissolved in ethyl acetate and water. The organic layer was separated, washed once with water and once with saturated sodium chloride solution, dehydrated with sodium sulfate, and filtered. The oily substance obtained by removing the solvent was stirred in 60 ml of pentane for 15 minutes, and then dried in vacuo to give 2.35 g of a powdery target compound.

D) (S)-(trans) -3-amine-4-ethynyl-2-oxo-1-azetidinesulfonic acid

404 mg of (S)-(trans) -4-ethynyl-2-oxo-3-[(triphenylmethyl) amino] azetidine and 560 mg of pyridine-sulfur trioxide complex were placed in a 25 ml flask. After filling the flask with nitrogen, 4.0 ml of anhydrous pyridine was added and the mixture was heated at 80 ° C. to 85 ° C. for 3 hours. This mixture was added to a mixture obtained by rapidly stirring 4.0 ml of concentrated hydrochloric acid, 50 ml of water, and 50 ml of ethyl acetate. Sodium carbonate was added to the mixture to adjust the pH to 3.15, and the aqueous layer was separated and then extracted once with ethyl acetate. The combined organic layers were washed once with saturated sodium chloride solution, dehydrated with sodium sulfate and filtered. The solvent was removed in vacuo, and the resulting foamy substance was dissolved in 10 ml of dichloromethane, followed by addition of 8 ml of formic acid (98%), after 15 minutes the mixture was concentrated to 4 ml, and then 10 ml of dichloromethane was added thereto. The solid obtained was suspended in solution. This suspension was filtered to give 100 ml of the target compound in the solid phase. This solid was melted at 1800 ° C. or higher to be discolored.

Example 192

[3S- [3α (Z), 4β]]-3-[[(2-amino-4-thiazolyl) (methoxyimino) acetyl] amino] -4-ethynyl-2-oxo-1-azetidine Sulfonic acid, potassium salt

85 mg (Z) -2-amino-α- (methoxyimino) -4-thiazole acetic acid 100 mg N-hydroxybenzotriazole monohydrate and 113 mg of dicyclohexylcarbodiimide were placed in a 10 ml flask. The flask was filled with nitrogen and cooled in an ice water bath. Subsequently, 0.5 ml of dimethylformamide was further added thereto. Here, 95 mg of (S)-(trans) -3-amino-4-ethynyl-2-oxo-1-azetidinesulfonic acid (see Example 191) was added 1.0 mg of dimethylformamide and triethylamine in solid form. Added with 56 μl. The cooling bath was removed and the mixture was stirred for 22 hours. 3 ml of acetone were added, then the resulting solids were distilled off and washed with 4 ml of acetone. All solvents were removed in vacuo and the residue was dissolved in 5 ml of methanol, then 162 mg of potassium sulfate perfluorobutane was added and dissolved. After the solution was left to stand, the resulting solid was isolated by centrifugation to obtain 68 mg of the target compound having a melting point of 230 ° C. or higher.

Example 193

(S) -3-[[[((2,5-dichlorophenyl) thio] acetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

100 mg of 3-amino-2-oxo-1-azetidinesulfonic acid was dissolved in 2 ml of anhydrous dimethylformamide and 0.083 ml of triethylamine, followed by 123 mg (0602 mmol) of 2,5-dichlorophenylthioacetic acid, N-hydride 81 mg of oxybenzotriazole hydrate and 124 mg of disiplohexylcarbodiimide were added and stirred at room temperature for 2 hours and then at 5 ° C. for 2 days. The solvent was removed in vacuo, the residue was dissolved in water, filtered through celite and the filtrate was washed with ethyl acetate. The aqueous layer was mixed with dichloromethane, to which 612 mg of tetrabutylammonium disulfide was added, followed by the addition of 1N sodium potassium hydroxide solution to adjust the pH to 3. After extraction three times with dichloromethane all, the extracts were combined and dehydrated with Na ₂ SO ₄ , and the solvent was removed in vacuo to give an oily material. The solution obtained by dissolving this oily substance in acetone was added to a solution of 612 mg of potassium sulfate perfluorobutane dissolved in acetone to precipitate the product. A small amount of ether was added thereto, filtered to obtain a solid, which was washed several times with acetone and dried to give 206 mg of powder.

Analysis of C ₁₁ H ₉ N ₂ O ₅ S ₂ Cl ₂ K

Theoretical value: C 31.21; H 2.14; Found: C 27.90; H 2.11;

N 6.62; Cl 16.75 N 5.84; Cl 18.04

Example 194

(3S-trans) -3-[[[((2,5-dichlorophenyl) thio] acetyl] amino] -4-methyl-2-oxo-1-azetidinesulfonic acid, potassium salt

250 mg of (3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid (see Example 139) were dissolved in 2 ml of dimethyl formamide, together with 193 μl of triethylamine. To this, 285 mg of 2,5-dichlorophenylthio acetate, 213 mg of N-hydroxybenzotriazole hydrate, and 287 mg of dicyclohexylcarbodiimide were added. After stirring for about 16 hours at room temperature, the mixture is filtered and the solvent is removed in vacuo. The residue was dissolved in water, filtered and the filtrate was washed with ethyl acetate, layered by the addition of dichloromethane and tetrabutylammonium disulfide (4.2 mmol) was added. The mixture was extracted three times with dichloromethane, and the extracts were combined, dehydrated with Na ₂ SO ₄ , and the solvent was removed in vacuo to obtain 920 mg of an oily substance. 945 mg of potassium sulfate perfluorofuran dissolved in acetone was added to a solution of the oily substance dissolved in acetone. The solid precipitated slowly, which was combined and washed twice with ether and dried to give 306 mg of powder. Chromatography was performed with HP-20 resin (100 ml column) and eluted with 20% acetonitrile: 80% water to obtain the desired compound, which was crystallized after evaporation in a water: methanol mixture. The residue was titrated with acetone to give 233 mg of powder having a melting point of 212-213 ° C (decomposition).

Analysis of C ₁₂ H ₁₁ N ₂ O ₅ Cl ₂ S ₂ K

Theoretic value: C 32.95; H 2.54; Found: C 32.91; H 2.60;

N 6.41; Cl 16.21; N 6.42; Cl 16.50;

S 14.66 S 13.77

[Examples 195 ~ 196]

(3S-trans) -3-amino-4-methyl-2-oxo-1-azetidinesulfonic acid is used instead of (3S-cis) -3-amino-4-methyl-2-oxo-1-azetidine sulfonic acid An acid described in Column I was used instead of (Z) -2-amino-α- (methoxy imino) -4-thiazoleacetic acid and the compound described in Column II was obtained according to the method of Example 138.

Example 197

[3S (R ^* )]-3-[[[((aminooxoacetyl) amino] (4-hydroxyphenyl) azetyl] amino] -2-oxo-1-azetidinesulfonic acid, potassium salt

(R)-[(amino instead of (Z) -2-amino-α-[[2- (diphenylmethoxy) -1,1-dimethyl-2-oxoethoxy] amino] -4-thiazole acetic acid It carried out according to the method of Example 28 using oxoacetyl) amino] (4-hydroxyphenyl) acetic acid, and obtained the target compound of melting | fusing point 128 degreeC (decomposition).

Example 198

[3S] -3-[[2-amino-4-thiazolyl) [[[3- (2-furanylethylene) amino] -2-oxo-1-imidazolidinyl] carbonyl] amino] acetyl] Amino-2-oxo-1-azetidinesulfonic acid, potassium salt

(R) -2-amino-α-[[[3- (2-furanylmethylene) amino] -2-oxo-1-imidazolidinyl] carbonyl] amino] -4- instead of aminothiazole acetic acid It carried out according to the method of Example 6 using thiazole acetic acid, and obtained the target compound of 250 degreeC or more of melting | fusing point.

Example 199

Biological Manufacturing of EM5117

9ℓ fermentation

Crobacterium bioraceum SC11, 378 A.T.C.C No. 31,32 was incubated in the following sterile agar medium (A).

Before sterilization at 121 ° C. for 30 minutes, the pH was adjusted to 7.3.

The microbial surface growth rings were inoculated in three 500 ml Erlenmeyer flasks each containing 100 ml of the following sterile medium (B).

Before sterilization at 121 ° C. for 15 minutes, the pH was adjusted to 7.0. The flask was incubated at 25 ° C. for about 24 hours on a rotary shaker (300 rpm: 2 inch stroke).

After appropriate incubation as described above, 1% (volume / volume) transition was transferred from growth culture flasks to 100 500 mm Erlenmeyer flasks each containing 100 ml of the following sterile medium (C).

Prior to sterilization at 121 ° C. for 15 minutes, the pH was adjusted to 7.0.

After inoculation, the flasks were incubated at 25 ° C. for about 18-24 hours on a rotary shaker (300 rpm: 2 inches Stork). At this time, the contents of the flask was taken out, and the fermentation broth was centrifuged to obtain about 9 L of the supernatant fermentation broth.

250ℓ fermentation

Agar Slope Medium (A) on Chromobacterium Bioraceum SC11,378 A.T.C.C. The surface growth rings incubated in No. 31,322 were inoculated in five 500 ml Etlenmeyer flasks each containing 100 mmol of sterile medium (B). The contents of this flask were then incubated on a rotary shaker (300 rpm: 2 inch stroke) at 25 ° C. for about 24 hours. After appropriate incubation, the 1% (volume / volume) transition was transferred from the growth culture flasks to five 4 liter Erlenmeyer Prasks, each containing 1.5 liters of the following sterile medium B, as described above. After inoculation, the flasks were incubated at 25 ° C. for about 24 hours on a rotary shaker (300 rpm: 2 inch stroke). As described above, after moderate incubation, the 1% (volume / volume) transition was transferred to a stirrer equipped with a fermentation tank containing 250 ml of sterile medium (C). After inoculation, fermentation was continued under the following conditions (temperature 25 ° C., pressure 10 psig; aeration 10 cfm; stirring 155 rpm). It was necessary to add yukon as an antifoaming agent. Fermentation was terminated after about 18-24 hours. HCl was added to the fermentation broth to adjust the pH to 5.0, and the contents of the fermentation broth in the fermentation tank were centrifuged to give about 230 L of supernatant fermentation broth.

Isolation and Purification

The fermentation broth obtained from 250 L fermentation was adjusted to pH 5 by adding sulfuric acid, filtered using 3-5% diatomaceous earth (Celtite), and the fermentation filtrate was twice with 30 L of 0.005 M cetyldimethyl benzyl ammonium chloride in methylene chloride. Extracted.

The combined phases at the bottom were extracted with 6 l of 0.05 M sodium iodide (adjusted to pH 5 with acetic acid). The lower phase was discarded and the upper phase was concentrated to 500 ml in vacuo.

This concentrate was extracted with 400 ml of n-butanol. The upper phase was discarded and the lower phase was concentrated and dried in vacuo. The residue was dissolved as much as 150 ml of methanol as much as possible. The insoluble material was discarded and the methanol solution was concentrated to dryness in vacuo to give 28.6 g of crude antibiotic.

This crude product was dissolved in 10 ml of methanol-water (1: 1), chromatographed with a 500 ml column of dextrangel (Sephadex G-10) crosslinked in the same solvent mixture as above, and the rate of 2 ml / min was eluted. 20 ml fractions were collected. The active fractions (19-26) were combined and concentrated in vacuo. 5.23 g of the residue was mixed with 50 ml of methanol. Insoluble material was filtered off and the filtrate was concentrated in vacuo.

5.0 g of the residue was dissolved in 10 ml of 0.01 M sodium phosphate buffer (pH5) and filled with the same buffer to equilibrate a equilibrated DEAE cellulose (Whatman DE 52 cellulose) column at 0.01 ml sodium phosphate buffer (pH 5) at a rate of 5 ml / min. Elution was carried out with a gradient of 4 L and 4 L of 0.1 M sodium phosphate buffer (pH 5) followed by the collection of 20 ml fractions. The sulfur fractions (192-222) were combined and concentrated in vacuo, the methanol insolubles were removed and washed well with methanol. The solvent was removed to give 576 mg.

576 mg of the residue was dissolved in 4 ml of water, and about 1 ml of 0.1 N sodium hydroxide was added thereto to adjust the pH to 5. The solution was chromatographed with an alkylated and separated Tranxel (Sepadex LH-20) column in water, eluted at a rate of 1 ml / min, and the 10 ml fractions were collected. The active fractions (38-44) were combined and concentrated to give 459 mg of residue.

348 mg of the residue was dissolved in water and the solution was placed in a macroreticular styrene-divinylbenzene copolymer resin (Diaion HP 20AG) column. The column was first washed with methanolic potassium hydroxide, methanol, methanolic hydrogen chloride, methanol and water and then filled with rules. This column was eluted with water at a rate of 1 ml / min and 10 ml fractions were collected. The active fractions (36-43) were combined and then concentrated to give 186.4 mg of material. 100 mg of 459 mg of the remaining mule obtained from the previous step was taken again and chromatographed in the same manner to give 51.5 mg of the material. As revealed by TLC and NMR spectra, at this stage 186.4 mg of substance and 51.5 mg of substance were almost pure EM5117.

The 186.4 mg portion of EM5117 described above was dissolved in water, passed through an ion exchange resin (Dowex 50W-X2, 100-200 mesh, K ⁺ type) column and washed with two phase volume of water. The effluent was concentrated to give 189.0 mg of crystalline solid. This material was dissolved in 0.38 ml of water and then recrystallized by addition of 3.42 ml of methanol. The resulting mixture was cooled with ice and filtered to give 145 mg of crystals. In this manner, the mixture was recrystallized twice with water-methanol (1: 9) twice to obtain 95.9 mg of EM5117 potassium salt having a melting point of 194 ° C (decomposition).

The following fermentation broths are effective for the preparation of EM5117 and may be used in place of the medium (B) and (C) in the above examples.

Badge D

Badge E

Badge F

Badge G

Badge H

Badge I

Badge J

Badge K

Badge L

Badge M

Badge N

Example 200

Biological Preparation of EM5210

Gluconobacter SC11,435 A.T.C.C. No. 31581 was incubated in the following agar agar medium (A).

Before sterilization at 121 ° C. for 30 minutes, the pH was adjusted to 7.3.

Surface growth rings incubated with Gluconobacter SC11,435 in agar slope medium (A) were inoculated in three 500 ml Erlenmeyer flasks each containing 100 ml of the following sterile medium (B).

Before sterilization at 121 ° C. for 15 minutes, the pH was adjusted to 7.3.

After inoculation, the flasks were incubated at 25 ° C. for about 24 hours on a rotary shaker (300 rpm: 2 inch stroke). After appropriate incubation as described above, the 1% (volume / volume) transfer was transferred from growth culture flasks to 100 500 ml Erlenmeyer flasks each containing 100 ml of the following sterilized medium (C).

The medium was sterilized at 121 ° C. for 15 minutes.

After inoculation, the flasks were incubated at 25 ° C. for 18 hours on a rotary shaker (300 rpm: 2 inch stroke). At this time, the contents of the flask was taken out, and the fermentation broth was centrifuged to obtain about 9 L of the supernatant fermentation broth.

Isolation and Purification (Small)

The active material from 10 l of the fermentation broth was adsorbed onto 500 g of a strong basic anion exchange resin column treated with a quaternary ammonium group adsorbed on a styrene-divinyl benzene copolymer lattice (Dowex AGI = X2 (Cl ⁻ )), followed by washing with water. Eluted with 5% sodium chloride in an aqueous 0.01 M NaH ₂ PO ₄ solution, which was concentrated in vacuo.

The residue was adsorbed onto 250 g of activated carbon washed with water and eluted EM5210 with methanol-water (1: 1). The active fractions were combined and concentrated in vacuo to afford crude EM5210.

Division EM5210 1ℓ of water and 2M acetic acid pyridium (pH 4.5) strong base anion exchange resin with a linear gradient using 1ℓ (Bio Rad AG 1X2 (Cl -.)) Was subjected to chromatography Griffey with 280ml column. The active fractions were combined and concentrated in vacuo.

Partially purified EM5210 was further purified by gel filtration on a 500 ml column of dextrangel (Sephadex G-10) that had been subjected to water using eluent as the eluent. The active fractions were combined and concentrated to give 26 mg EM5210. The potassium salt of EM5210 was prepared by passing EM5210 through a potassium type cation exchange resin column (Dowex 50-X2).

Isolation and Purification (Large Scale)

Glucono bakteo SC11,435 of 250ℓ fermentation broth (pH 3.7) strong base anion exchange resin in the fermentation broth filtrate ^{(Dowex 1-X8 (Cl -} )) was adsorbed to 10.8kg. The resin was washed with water and then eluted with 5% sodium chloride in 0.01 M sodium dihydrogen phosphate. The active fractions were combined and concentrated to a small volume. The precipitated salts were removed and the filtrate was desalted by passing through activated carbon columns (20-40 mesh Darco 1.1 kg) in water. The column was washed with water and EM5210 eluted with methanol-water (1: 1). The active fractions were combined and concentrated. The residue was dissolved in 16g water, a strongly basic ion exchange resin (Bio Rad AG1-X2 (Cl -.)), 200 ~ 400 mesh) performing heart, 1ℓ of water and the 0.01M phosphate by column chromatography Griffey 600ml 2 hydrogen Eluted with a linear gradient using 1 L of 10% sodium chloride in sodium. The active fractions were combined and concentrated in vacuo until a small volume, then the precipitated salts were filtered off. This filtrate was eluted with water using a macroreticular styrene-thivinylbenzene copolymer resin column. The active fractions were combined and concentrated to a small volume and then lyophilized to give 120 mg of the sodium salt of EM5210.

In order to convert the sodium salt to lithium salt, an ion exchange resin (Dowex 50W-X2, lithium type) column was used. 100 mg of sodium salt was dissolved in 0.5 ml of water and eluted with water using a column. The active fractions were combined and directly lyophilized to obtain 95 mg of lithium salt of amorphous solid EN5210.

The free acid (molecular salt) of EM5210 was made into H ⁺ type by passing the base salt of EM5210 through the column of the weak acid ion exchange resin. For example, about 2.5 mg of lithium salt may be used in Bio. Rad Bio. Elution with water using a Rex 70 (H ⁺ ) column yielded 1.45 mg of free acid (intramolecular salt).

Chemical Properties of EM5210

1) ninhydrin positive

2) Positive of two major ninhydrins by paper chromatography (Whatman No. 1, butanol-acetic acid-mol (5: 1: 14)) by acid hydrolysis (hydrolysis for 16 hours with 6N HCl at 115 ° C.). Quenching with fluorescence excited by spot and UV showed one weak ninhydrin positive spot. The two main ninhydrin positive spots were D-glutamic acid and D-alanine.

Physical Properties of EM5210

1) UV spectrum of sodium salt in water: terminal absorption

2) Main peaks of lithium salt in IR-KBr: 1770, 1640, 1530, 1384, 1242 and 1051 cm ^-1 ,

3) Chemical variation of lithium salts in NMR-water (ppm downfield from TSP): 1.40 (d, J = 7 Hz)

2.14 (m) 3.73 (t, J = 6 Hz)

2.44 (m) 3.94 (s)

3.49 (s) 4.28 (m)

Beneficiation of free acid (intramolecular salt) in water at 24 ° C. (C = 0.15%, pH2.7):

λ (nm) [α] λ (nm) [α]

589 + 73 ° 436 + 159 °

578 + 79 ° 365 + 263 °

546 + 91 °

The following fermentation media are effective for preparing EM5210 and may be used in place of the mediums (B) and (C) described herein.

Badge D

The pH was adjusted to 7.0 before sterilization at 121 ° C. for 15 minutes.

Badge E

Sterilize at 121 ° C. for 15 minutes.

Badge F

Adjust pH to 6.0 before sterilization at 121 ° C. for 15 minutes.

Badge G

Sterilize at 121 ° C for 30 minutes.

Badge H

Sterilize at 121 ° C for 15 minutes

Badge I

Sterilize at 121 ° C. for 30 minutes.

Biological effect

The following method is to measure the super inhibitory concentration (hereinafter referred to as MIC) of β-lactam of the present invention.

The fermentation broth was inoculated in a ring in vitro of microorganisms obtained from BHI (Difco) agar slope medium to cultivate the microorganisms under test in about 15-20 ml of antibiotic assay fermentation broth (Difco). Inoculated test tubes were incubated at 37 ° C. for 18-20 hours. These cultures were estimated to contain 10 ⁹ colony forming units (hereinafter referred to as CFU) per ml. This culture was diluted to about 1:10 to obtain a final inoculation concentration of 10 ⁴ CFU and diluted with K-10 fermentation broth.

This compound was dissolved in a suitable diluent at a concentration of 1,000 μg / ml. Diluted two more times with K-10 fermentation broth to 1,000 μg / ml to 0.5 μg / ml. 1.5 ml of each dilution was placed in a square Petri dish, and 13.5 ml of K-10 agar was added thereto. The final concentration of the drug in the agar was 1,000 μg / ml to 0.05 μg / ml. Agar-containing microbial growth control plates were made and inoculated before and after the test plates. Microorganisms were inoculated on the agar surface of each plate with a Denley Multipoint Inoculator (inoculated with approximately 0.001 ml of microorganisms) to obtain a final inoculum concentration of 10 ⁴ CFU on the agar surface.

The plate was incubated at 37 ° C. for 18 hours and MIC was measured. MIC is the lowest concentration of compound that inhibits the growth of microorganisms.

K-10 fermentation broth is a yeast beef fermentation broth containing the following ingredients.

Beef Extract 1.5g Dextrose 1.0g

Yeast extract 3.0g distilled water up to 1ℓ

Peptone 6.0g

K-10 agar

Beef Extract 1.5g Dextrose 1.0g

Yeast Extract 3.0g Agar 15.0g

Peptone 6.0g

About 1ℓ of distilled water

The following tables tabulate the results of testing the β-lactam of the present invention against various bacteria. The numbers following each strain name are the microbial collection numbers of applicants E. R. Swim & Sons, Inc., Princeton, New Jersey, USA. The symbol (-) in the table means that the compound under test was ineffective against certain bacteria at 100 μg / ml. "N.T" means not tested.

Claims (1)

When the amino substituent R ¹ of the formula (II) is hydrogen or acyl, sulfonated the β-lactam of the formula (II), or when the amino substituent R ¹ is an amino protecting group, the β-lactam of the formula (II) A process for preparing the β-lactam derivative of the following formula (I) characterized by introducing acyl after sulfonation or sulfonating after acyl.

In each formula above, R ₁ is hydrogen or acyl group in formula (I), and hydrogen, acyl group or amino protecting group in formula (II), wherein the acyl group is selected from the group consisting of the following (a) to (g). (a) aliphatic groups of the formula

Wherein R ₅ is alkyl substituted with alkyl, alkylthylo or cyanomethyl groups. (b) a carbocyclic aromatic group of the formula

Wherein n is 0 or 1, R ₆ , R ₇ and R ₈ are each hydrogen, halogen, alkoxy having 1 to 4 carbon atoms, or aminomethyl, and R ₉ is hydroxyl, carboxyl salt, protected carbon Compound, sulfo salt, azido, halogen or [(alkylthio) thioxomethyl] thio. (c) heteroaromatic groups of the formula

Where n is 1 and R ₁₀ is

(d) the following structural formula

Where R ₁₁ is

or

And R ₁₂ is alkyl or amino. (e) the following structural formula:

Where R ₁₁ is

or

And, R ₁₃ is hydrogen, alkyl, cycloalkyl, alkylamino-carbonyl, or hydrogen, alkylthio, carboxyl, carboxylate salts, alkoxy carbonyl, phenyl, ethoxy, carbonyl, diphenyl methoxycarbonyl, hydroxy Alkyl substituted with either (phenylmethoxy) phosphinyl or dialkoxyphosphinyl substituents. (f) the following structural formula:

Where R ₁₁ is

R ₁₄ is amino; Alkylamino or

to be. (g) the group of the formula

Where R ₁₁ is

And R ₁₅ is hydrogen or alkylsulfonyl. R ₂ is hydrogen or C ₁ -C ₄ alkoxy. R ₃ and R ₄ are the same or different and are each hydrogen, alkyl, cycloalkyl, phenyl or substituted phenyl, or one of R ₃ and R ₄ is hydrogen and the other is alkoxycarbonyl, alken-1-yl, alkyne-1 -Yl, 2-phenylethenyl or 2-phenylethynyl, provided that when R ₃ and R ₄ are the same or different and each of them is substituted phenyl, then one of R ₃ and R ₄ is hydrogen and the other Not alken-1-yl or 2-phenylethynyl. M ⁺ is hydrogen or a cation.