HUP0000288A2 - Azetidinone derivatives that inhibit the enzymatic activity of PSA, their intermediates and their production, as well as pharmaceutical preparations containing the compounds - Google Patents

Abstract

The subject of the present invention is new azetidinone derivatives of formula (I) - where R1 in the formula is an optionally substituted aryl or alkyl group or a heterocyclic group, R2 is an alkenyl, aryl or heterocyclic group, and R3 is a heteroring, an esterified carboxyl group or a thiocarbamoyl group -, certain intermediates products and the production processes of the above azetidinone derivatives, as well as pharmaceutical preparations containing the compounds. The azetidinone derivatives according to the present invention are suitable for inhibiting the enzymatic activity of prostate-specific antigen (Prostate-Specific Antigen, PSA), as well as prostate cancer (Pca), metastases of aPca, benign prostatic hyperplasia (BPH) and breast cancer (Bc) for treatment. HE

Description

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PSA ENZYMATIC ACTIVITY INSTRUMENTS

The present invention relates to novel azetidinone derivatives, certain intermediates, and processes for preparing said azetidinone derivatives, and pharmaceutical compositions containing said compounds. The present invention also relates to the use of said azetidinone derivatives for inhibiting the enzymatic activity of prostate-specific antigen (PSA) and for treating prostate cancer (PCa), PCa metastases, benign prostatic hyperplasia (BPH) and breast cancer (Be). Another aspect of the invention relates to the treatment of PCa, PCa metastases, BPH and Be with any PSA inhibitor compound.

Prostate cancer is the second most common cancer in American men. According to Wingo et al. [P. A. Wingo, et al., CA Cancer J, Clin., 41, (1), 8-30, (1995)], in the United States in 1995 there were 244,000 new cases diagnosed and 40,000 cancer-related deaths. This statistic represents 36% of all cancer cases in men and 13% of cancer-related deaths in men.

There is currently no known effective cure or preventive treatment for prostate cancer. When the cancer is in its early, hormone-dependent stages, it is usually treated by removal of the testicles or chemical sterilization, and sometimes the androgen flutamide is used. Prostate cancer is

In its later stages, the disease becomes hormone-independent and metastasizes widely, usually through the skeleton. In the advanced stages of the disease, radiation therapy is used to relieve pain and slow metastasis in the irradiated area, but there is no effective method to reverse the progression of prostate cancer in the late stages of the disease. Thus, prostate cancer is not only a relatively common disease, but its treatment is also ineffective once the disease has reached the hormone-independent stage.

Benign prostatic hyperplasia (BPH) is another disease of the prostate. BPH is a noncancerous condition characterized by excessive cell growth in the prostate gland. The enlarged prostate blocks the urethra, often causing symptoms such as difficulty urinating, a slow or interrupted stream of urine, frequent urination, and dribbling after urination. Other symptoms not directly related to urination may include hernias, hemorrhoids, changes in bowel habits, and other consequences of increased abdominal pressure and straining during urination. It is estimated that about 80% of men over the age of 50 have BPH, and 20% of men over the age of 80 require surgery to reduce the resulting symptoms. P. Narayan and R. Indudhara, The Western Journal of Medicine, 161, 495 (1994).

Breast cancer is another devastating disease for which better treatments are desperately needed. Breast cancer is a leading cause of death in women, as well as a major cause of disability, psychological distress, and economic loss. A large number of women with this disease eventually die from the disease, either directly or as a result of complications such as metastasis, deterioration of general health, or side effects of treatment-related interventions (such as surgery, radiation therapy, or chemotherapy). The long-term prognosis for breast cancer patients, even with the best combination of treatments (surgery, radiation therapy, and/or chemotherapy), is variable and is very poor in the presence of metastasis.

Prostate-specific antigen (PSA) is considered a causative factor in prostate cancer, its metastases, benign prostatic hyperplasia, breast cancer, and its metastases. PSA is an androgen-dependent, 28 kDa glycoprotein (protein containing sugar groups) produced almost exclusively in the endothelium of the prostate, and is most commonly found in seminal fluid. [A. F. Prestigiscomo et al. J. Urol., 152, 1515-9 (1994) and G. G. Klee et al. Urology, 44, 76-82 (1994)].

PSA is currently used as a serum marker (a substance that indicates a pathological condition and can be detected in the blood serum) in the diagnosis and monitoring of prostate cancer. In the last decade, a large amount of data has accumulated on the association between increased serum PSA concentrations and prostate cancer. In healthy men, 85-95% of the PSA present in the circulation is bound to either α-antichymotrypsin (ACT) or α- ₂ ^- macroglobulin. Bound PSA is enzymatically inactive or unable to interact with its physiological substrate. In patients with prostate enlargement and prostate cancer, the ratio of bound to free PSA in the blood serum varies. PSA has also been discovered in extracts from breast cancer and on this basis

It is proposed to be used as a breast cancer predictor. H. Yu et al., Cancer Research, 55, 2104-10 (1995).

In addition, PSA is thought to have a physiological role in male reproductive function, as this serine protease enzyme has activity in the liquefaction of seminal plasma. PSA has recently been shown to be expressed in tissues other than the prostate and to mediate proliferative and metastatic responses in cell cultures. A critical review of these recently published preclinical data supports a possible pathophysiological role for PSA in the promotion and metastasis of prostate cancer and prostatic hyperplasia as a growth factor mediator. There is also evidence that the serum concentrations of PSA and IGFBPs (insulin growth factor binding proteins) are correlated in patients with prostate cancer. H. Kanety et al., J. Clin. Endocrinol. Metab., 77, 229-33 (1993). PSA has been shown to degrade certain IGFB proteins, which may contribute to prostate cell growth, leading to prostate enlargement and prostate cancer. P. Cohen et al., J. Endocrinol,, 142, 407-415 (1994). PSA has also been shown to directly stimulate the growth of prostate endothelial cells in vitro (observed in a test tube). In addition, the ability of certain IGFB proteins to inhibit the growth of breast cancer cells is consistent with the hypothesis that PSA plays a similar role in the treatment of breast cancer as it does in the treatment of prostate cancer.

The present invention relates to a compound of general formula I, wherein the

The group of general formula R ¹ means aryl, aryl-(1-6C alkylene)-group, in which the aryl group, or the aryl-(1-

The ring of the alkylene group may be substituted with one or more groups selected from the group consisting of halide, alkyloxy, methoxycarbonyl, phenyl, alkyloxy, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene; phthalimido, or one of the groups of formula (a) to (e), wherein the

R ⁴ is a hydrogen atom, a C 1-6 alkyl, aryl, or heterocycle; wherein the aryl or heterocycle may be optionally substituted with one or two groups selected from the following: halide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene;

R ⁵ is a hydrogen atom, a C 1-4 alkyl, a C 3-7 cycloalkyl, a C 1-4 alkyloxycarbonyl, a phenyl, or a naphthyl group, wherein the phenyl or naphthyl group may be optionally substituted with one or two groups selected from the following: C 1-4 alkyl, C 1-4 alkyloxy, halide, hydroxyl, cyano, carbamoyl, amino, mono(C 1-4 alkylamino, di(C 14 alkylamino, C 1-4 alkylsulfonylamino, and nitro groups;

R ⁶ represents a hydrogen atom, a C 1-4 alkyl group optionally substituted with one or two groups selected from the following: hydroxyl, protected carboxyl, carbamoyl, benzylthio or C 1-4 alkylthio groups; or a phenyl group optionally substituted with one or two groups independently selected from the following:

with group: C1-4 alkyl group, C1-4 alkyloxy group, halide, hydroxyl, cyano, carbamoyl, amino, mono(C1-4 alkyl)amino, di(C1-4 alkyl)amino and nitro group, or C1-4 alkyloxycarbonyl group;

R ⁷ is a C 1-4 alkyloxycarbonyl, benzyloxycarbonyl, benzoyl group, wherein the phenyl group of the benzyloxycarbonyl or benzoyl group may be optionally substituted with one or two groups independently selected from the following: C 1-4 alkyl, C 1-4 alkyloxy, halide, cyano, nitro, amino, carbamoyl, hydroxyl, mono(C 1-4 alkyl)amino and di(C 1-4 alkyl)amino groups; the groups of general formula R ⁸ and R ⁹ independently represent a hydrogen atom, a C 1-5 alkanoyloxy, a benzoyloxy group, which may be substituted by one or two groups independently selected from the following: a C 1-4 alkyl, a C 1-4 alkyloxy, a halide, a cyano, a nitro, an amino, or a C 1-4 alkyloxycarbonyl group; a benzyloxy, a diphenylmethoxy, or a triphenylmethoxy group; with the proviso that only one of the groups of general formula R ⁸ and R ⁹ may represent a hydrogen atom;

R ² is a C 2-6 alkenyl, aryl, or heterocycle, wherein the aryl or heterocycle is optionally substituted with one or two groups independently selected from the group consisting of halide, n-oxide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxyethylene, and hydroxymethylene;

-CH ₂ CH ₂ R ¹⁰ is a group of general formula, in which the

R ¹⁰ is a carboxyl, aryl or heterocycle, wherein the aryl or heterocycle may be optionally substituted with one or two groups independently selected from the group consisting of halide, n-oxide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene and hydroxymethylene;

-CH = C(R ¹² )-R ¹¹ group, in which the

R ¹¹ is hydrogen or phenyl, and R ¹² is nitrile, aryl or heterocycle, wherein the aryl or heterocycle may be optionally substituted with one or two groups independently selected from halide, n-oxide, C1-6 alkyloxy, methoxycarbonyl, phenyl, C1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene; with the proviso that when R ¹¹ is phenyl, R ¹² is aryl; or

-C=CR ¹³ is a group of general formula, in which the

The group of general formula R ¹³ represents an aryl group or a heterocycle, where the aryl group or the heterocycle may be optionally substituted with one or two groups, which are independently selected from the group consisting of

selected from the following: halide, N-oxide, C1-6 alkyloxy, methoxycarbonyl, phenyl, C1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene; furthermore, the group of general formula R ³ represents a heterocyclic ring, CO ₂ R ¹⁴ , or a group of general formula (f), wherein the

R ¹⁴ is a C 2-6 alkenyl, C 1-6 alkyl, C 1-6 alkyl substituted with halogen, C 3-7 cycloalkyl, substituted C 3-7 cycloalkyl, aryl-substituted C 1-6 alkyl or heterocycle, wherein the aryl group or the heterocycle or the ring of the aryl-substituted C 1-6 alkylene group may be optionally substituted with one or two groups independently selected from the following: halide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene;

R ¹⁵ is an aryl group or a heterocyclic substituted C 1-6 alkylene group, wherein the ring of the aryl group or the heterocyclic substituted C 1-6 alkylene group may be optionally substituted with one or two groups independently selected from the group consisting of halide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, trifluoromethyl, and hydroxymethylene;

and salts and solvates of these compounds.

The present invention provides a method for inhibiting the proteolytic activity of prostate-specific antigen (PSA) using a compound of formula I.

The present invention further provides methods for treating prostate cancer, metastases thereof, benign prostatic hyperplasia, breast cancer and metastases thereof by administering to a mammal in need of said treatment a prostate-specific antigen inhibitor. Preferred prostate-specific antigen inhibitors are compounds of formula I. This aspect of the present invention is based on the discovery that PSA inhibitory agents are effective in treating the aforementioned diseases. Those skilled in the art can readily determine whether a compound has PSA inhibitory activity using known methods.

Another aspect of the present invention is processes and intermediates useful for the preparation of compounds of formula I, as well as pharmaceutical compositions containing compounds of formula I.

Unless otherwise indicated, the terms and abbreviations used in this specification have the usual meanings. For example, the term “°C” means degrees Celsius; “N” means normal solution or normality; “mmol” means millimole; “g” means gram; “ml” means milliliter; “M” means molar solution or molarity; “MS” means mass spectrometry; “IR” means infrared spectroscopy; “NMR” means nuclear magnetic resonance spectroscopy.

The term "aryl group" as used herein means a group of 6 to 14 carbon atoms, containing one ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl and anthracyl).

aryl group is an unsaturated aromatic cyclic hydrocarbon group. If the group is substituted, the substituents may be in any position on the aryl ring(s) that is spherically (sterically) possible and that results in a stable compound. The aryl group may be optionally substituted with one or more groups. Examples of substituted aryl groups include 4-fluorophenyl, 4-chlorophenyl, 4-iodophenyl, 4-nitrophenyl, 4-methylphenyl, 4-methoxyphenyl, 4-ethoxyphenyl, 1-naphthyl, 2-naphthyl, 2-chloronaphthyl, 2,3-dichloronaphthyl, 2-iodonaphthyl, 3-iodonaphthyl, 2-fluoronaphthyl, 3-fluoronaphthyl, 2-methylnaphthyl, and 3-methylnaphthyl.

The term "substituted C3-7 cycloalkyl" as used herein refers to a C3-7 cycloalkyl group which may be substituted with one or two groups selected from halide or C1-6 alkyl groups. Examples of substituted cycloalkyl groups include 2-isopropylcyclohexyl, 5-methylcyclohexyl, 2-isopropyl-5-methylcyclohexyl, 4-chlorocyclohexyl, 3-chlorocyclohexyl, 4-iodocyclohexyl, 3-iodocyclohexyl, 3-chlorocyclopentyl, and 3-chlorocycloheptyl.

The term "heterocyclic group" as used herein means a monovalent saturated or unsaturated group containing a single (5- or 6-membered) ring or a fused ring system consisting of multiple rings (9- or 10-membered), containing up to 4 nitrogen atoms and/or up to 2 oxygen atoms and/or up to 2 sulfur atoms, and which forms a sterically feasible, stable compound. Examples of heterocycles include 2-thienyl or 3-thienyl, 2-furyl or 3-furyl, pyrrolyl, pyridyl, pyrimidyl, imidazolyl, pyrrolidinyl, piperidinyl, azepidinyl, indolyl, isoindolyl, quinolinyl, isoquinolinyl, benzothiazolyl, benzoxazolyl, benzimidazolyl, benzoxadiazolyl,

and benzofurazanyl. The heterocycle may be optionally substituted with one or two groups selected from the group consisting of C1-4 alkyl, C1-4 alkyloxy, halide, hydroxyl, and oxo. Examples of substituted heterocycles include pyridin-N-oxid-3-yl, pyridin-N-oxid-4-yl, and 4-methylpyridin-3-yl, and the like.

The term "C1-6 alkyl" as used herein refers to a branched or straight chain monovalent alkyl group having from one to six carbon atoms. Typical C1-6 alkyl groups include methyl, ethyl, n-propyl, isopropyl, η-butyl, isobutyl, sec-butyl, tert-butyl, η-pentyl, isopentyl, n-hexyl, 2-methylpentyl, and the like. The term C1-6 alkyl group also includes groups defined by a narrower range, such as C1-4 alkyl and C2-5 alkyl.

The term "C2-C6 alkenyl group" as used herein means a straight or branched, monovalent, unsaturated, aliphatic chain containing from two to six carbon atoms and having one double bond. Typical C2-C6 alkenyl groups include ethenyl (also known as vinyl), 1-methylethenyl, 1-methyl-1-propenyl, 1-hexenyl, 2-methyl-2-propenyl, 1-propenyl, 1-butenyl, 2-pentenyl, and the like.

The term "C1-6 alkyloxy" as used herein refers to a straight or branched chain C1-6 alkyl group attached to an oxygen. The oxygen atom connects the alkyl group to the parent molecule. Typical C1-6 alkyloxy groups include methoxy, ethoxy, propyloxy, isopropyloxy, butoxy, tert-butyloxy, pentyloxy, and the like.

The term "halogenated alkyl group having 1-6 carbon atoms" as used herein refers to a straight or branched alkyl chain having from one to six carbon atoms to which one or more halogen atoms are attached. Typical halogenated alkyl groups include 3-chlorobutyl, 4-chlorobutyl, 3-iodobutyl, 4-iodobutyl, 3-fluorobutyl, 4-fluorobutyl, and the like.

The term "C1-6 alkylene group" as used herein means a straight carbon chain containing from one to six carbon atoms and having two free valences. Typical C1-6 alkylene groups include ethylene, trimethylene, tetramethylene, and the like.

The term “aryl-substituted C1-6 alkylene group” as used herein refers to a C1-6 alkylene substituent group having a product group, in which the alkylene group is a straight chain, such as phenylethylene group and the like. The group is attached to the parent molecule through the alkylene moiety. The ring of the aryl-substituted C1-6 alkylene group may be optionally substituted with one or more groups. The substituents may be at any free position on the aryl ring.

The term "halo" or "halide group" as used herein means a fluoride, chloride, bromide, or iodide group.

The term "C1-4 alkyloxycarbonyl" as used herein means a C1-4 alkyloxy group attached to the parent molecule through a carbonyl group [-C(O)-(C1-4 alkyloxy)].

The term "benzylthio group" as used herein refers to a benzyl group (-S-CH ₂ -phenyl) attached to a sulfur atom. The group is attached to the parent molecule by the sulfur atom.

The term "carbamoyl group" as used herein means a group of the formula NH ₂ CO-.

The term "hydroxymethyl group" as used herein means a group of the formula -CH ₂ OH.

The term "N-oxide" as used herein refers to a radical oxygen atom attached to a nitrogen in a molecule.

The term "C1-5 alkanoyloxy" as used herein means a group of the formula (C1-5 alkyl)-C(O)O-. Examples of C1-5 alkanoyloxy include acetoxy, pivaloyloxy, and the like.

The term "treatment" as used herein refers to the preventive treatment of a named physiological condition, or the delaying of the onset of said physiological condition, or, if the existence of said condition can be determined, the amelioration or elimination thereof.

The compounds of the present invention, commonly referred to as azetidine derivatives, represent a new class of compounds that inhibit the proteolytic activity of prostate-specific antigen (PSA) (hereinafter referred to as "PSA inhibitors"). The exemplary compounds of the present invention inhibit the serine protease activity of PSA with increased selectivity over other serine protease enzymes [such as HLE, Human Leukocyte Elastase, tPA, tissue plasminogen activator, and thrombin].

The compounds of the present invention form solvates in known manner with suitable solvents. Favorable solvents for the formation of solvates include water, alcohols, tetrahydrofuran (THF), dimethylformamide (DMF), and dimethylsulfoxide (DMSO). Favorable alcohols include methanol and ethanol. Other suitable solvents may be selected based on the size of the solvent molecule. The formation of a suitable solvate is facilitated if the solvent is of small molecular weight. Typically, the solvate is formed during recrystallization or during salt formation. A useful reference for dealing with solvates is Sykes, Peter, A Guide to Organic Chemical Mechanisms, 6, 56 (1986).

The compounds of the present invention can also form salts with a variety of inorganic and organic acids. Typical acids that can be used for this purpose include sulfuric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, hypophosphoric acid, hydroiodic acid, sulfamic acid, citric acid, acetic acid, maleic acid, malonic acid, succinic acid, tartaric acid, cinnamic acid, benzoic acid, ascorbic acid, mandelic acid, p-toluenesulfonic acid, benzenesulfonic acid, methanesulfonic acid, trifluoroacetic acid, hippuric acid, and the like. Pharmaceutically acceptable, favorable salts are those formed with hydrochloric acid or acetic acid.

Although the compounds of formula I are effective PSA inhibitors, certain groups of compounds of formula I are more advantageous for said use. Accordingly, preferred embodiments of the present invention are those compounds of formula I wherein the

R ¹ is a benzyl, phthalimido, or (g) group, wherein

R ⁵ represents a hydrogen atom, a C 1-4 alkyl group, or a phenyl group;

R ⁶ represents a hydrogen atom, an isopropyl group, or a phenyl group;

The group of general formula R ² means a phenyl-C 2-C 4 alkenyl group, -CH ₂ CH ₂ R ¹⁰ , -CH=C(R ¹¹ )-R ¹² , or a group of general formula C^CR ¹³ , in which the

R ¹⁰ is a carboxyl or phenyl group;

R ¹¹ represents a hydrogen atom or a phenyl group;

R ¹² is cyano, phenyl, naphthyl, 2-furanyl, or 3-furanyl, pyridinyl, pyrimidinyl, or quinolinyl; and wherein the phenyl group may be optionally substituted with a C 1-4 alkyl, C 1-4 alkyloxy, or nitro group, and wherein the pyridinyl group may have an N-oxide; R ¹³ is phenyl;

R ³ is a heterocyclic group, CO ₂ R ¹⁴ , or (f) a group, wherein the heterocyclic group is pyridinyl, pyrimidinyl, 1,3,5-triazinyl, quinazolinyl or benzoxazolyl, and wherein said heterocyclic group is optionally substituted independently with one or two nitro, trifluoromethyl, C 1-4 alkyloxy or phenyl groups;

R ¹⁴ is a C 2-4 alkenyl, C 1-4 alkyl, C 1-4 halogenated alkyl, C 4-7 cycloalkyl, 2-isopropyl-5-methylcyclohexanyl, benzyl or phenyl group; wherein the phenyl group may be optionally substituted with a halide, C 1-4 alkyloxy, carbomethoxy or nitro group;

R ¹⁵ is a phenyl, naphthyl, 2-furanylmethyl, or 3-furanylmethyl group; wherein the phenyl group may be optionally substituted with one to four groups, which may independently be halogen or trifluoromethyl groups; and pharmaceutically acceptable salts and solvates of the above compounds. The most preferred embodiment of the present invention is the compound of formula (h), wherein

R ³ is a heterocyclic group, CO ₂ R ¹⁴ or (f) a general group of the formula, wherein the heterocyclic group is pyridinyl, pyrimidinyl, 1,3,5-triazinyl, quinazolinyl, or benzoxazolyl, and wherein said heterocyclic group is optionally substituted independently with one or two nitro, trifluoromethyl, C 1-4 alkyloxy, or phenyl groups;

R ¹⁴ is a C 2-4 alkenyl, C 1-4 alkyl, C 1-4 halogenated alkyl, C 4-7 cycloalkyl, 2-isopropyl-5-methylcyclohexanyl, benzyl or phenyl group;

R ¹⁴ is a C 2-4 alkenyl, C 1-4 alkyl, C 1-4 halogenated alkyl, C 4-7 cycloalkyl, 2-isopropyl-5-methylcyclohexanyl, benzyl or phenyl group; wherein the phenyl group may be optionally substituted with a halide, C 1-4 alkyloxy, carbomethoxy or nitro group;

R ¹⁵ is phenyl, naphthyl, 2-furanylmethyl, or 3-furanylmethyl; wherein the phenyl group may be optionally substituted independently with one to four halide or trifluoromethyl groups; and pharmaceutically acceptable acid addition salts and solvates of said compounds.

The numbering system set forth below is used in the present specification. The compounds of the present invention consist of an azetidinone central skeleton, with asymmetric carbon atoms at the 3- and 4-positions of said skeleton as illustrated in general formula (i). Thus, the compounds of the present invention may exist in the form of a single diastereoisomeric compound, a mixture of diastereoisomers, or a racemic mixture, all of which are within the scope of the present invention. The individual enantiomers may be separated by well-known classical resolution methods. A particularly useful reference describing such methods is Jacques et al., Enantiomers, Racemates, and Resolutions (John Wiley and Sons 1981). Suitable resolution methods include direct crystallization, optically active complexation, and recrystallization from an optically active solvent. Chrisey, L.A. Heterocycles, 267, 30 (1990).

Compounds of formula (i) may exist in two geometric isomers, the cis and trans isomers, or mixtures of these isomers. "Trans" isomers are those in which the substituent R ¹ is opposite or trans to the substituent R ^2. "Cis" isomers are those in which the substituent R ¹ is the same or cis to the substituent R ² .

The present invention therefore includes both R and S configurations for the 3- and 4-position groups. In this specification, the terms "R" and "S" are used in the conventional sense of organic chemistry to denote the configuration of an optically active carbon atom. See R. T. Morrison and R. N. Boyd, Organic Chemistry, pp. 138-139 (4th ed., Allyn & Bacon, Inc., Boston), and Orchin et al. Dictionary of Organic Chemistry, p. 126 (John Wiley and Sons, Inc.).

In a preferred embodiment of the present invention, the R ¹ substituent is cis-positioned with respect to the R ² substituent and has the (i1) configuration.

Also within the scope of the present invention are racemic mixtures of cis isomers [compounds of formula (i1) and (i2), mixtures of the 3S,4R and 3R,4S configurations]. In another embodiment of the present invention, the substituent group of formula R ¹ is trans-located with respect to the substituent group of formula R ² and has a 3R,4R configuration [formula (i3)], or a 3R,4S configuration [formula (i4)]. Also within the scope of the present invention are racemic mixtures of trans isomers (both 3R,4R and 3S,4S configurations). A third embodiment is a mixture of cis and trans isomers.

In a most preferred embodiment, the azetidinone backbone configuration is trans, preferably trans-3R,4R.

Racemic mixtures of cis and trans isomers, as well as mixtures of cis and trans isomers, are included within the scope of the present invention.

Although all combinations of stereochemical purity are contemplated by the present invention, it is preferred that the optically active carbon atoms be characterized by a single, absolute configuration. It will be apparent to those skilled in the art that when the substituent group represented by the general formula R ¹ is an oxazolidin-2-on-3-yl group, the carbon atom at the 4-position of that ring is asymmetric.

The following group of compounds is typical of the scope of the present invention:

- [4-(fluoro)-phenoxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)-ethen-1-ylazetidinone; trans-1-phenoxycarbonyl-3(2-phthalimido)-4-[2-(phenyl)-ethen-1-ylazetidinone; trans-1-[4-(nitro)benzyloxycarbonyl-3-(2-phthalimido)-4-[2 (phenyl)-ethen-1-ylazetidinone; 1-[(4-chlorobutyl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)-ethen-1-ylazetidinone;

trans-1-[(4-chlorobutyl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone; trans-1-[(4-iodobutyl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone; 1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone; trans-1-benzyloxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone; 1[(2-propen-1-yl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone; 1-[(2-isopropyl-5-methyl-hexyloxycarbonyl]-3-(2-phthalimido)-4-[2(phenyl)ethen-1-ylazetidinone; trans-1-[4-(methoxycarbonyl)phenoxycarbonyl]-3-(2-phthalimido)-4-[2(phenyl)ethen-1-ylazetidinone; trans-1-(4-(methoxy)phenoxycarbonyl]-3-(2-phthalimido)4-[2(phenyl)ethen-1-ylazetidinone; trans-1-(4-(methoxy)phenoxycarbonyl]-3-(2-phthalimido)4-[2(phenyl)ethen-1-ylazetidinone; trans-1-(1-propylenoxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-ylazetidinone; 1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(3-pyridyl)ethen-1-ylazetidinone;

1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(3-pyridyl)ethen-1-ylazetidinone hydrochloride; cis-1-phenoxycarbonyl-3-(2-phthalimido)-4[2-(phenylethylazetidinone; trans-1-phenoxycarbonyl-3-(2-phthalimido)-4-[2carboxyethylazetidinone; 1-phenoxycarbonyl-3-(2-phthalimido)-4-[2(diphenylethylazetidinone; cis-1-[4-(methoxycarbonyl)phenoxycarbonyl]-3-(2-phthalimido)-4-[2-(phenylethylazetidinone; cis-1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(phenylethylazetidinone;

trans-1-[4-(chloro)phenoxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone; cis-1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)-ethen-1-yl]azetidinone; cis-1-phenoxycarbonyl-3-(2-phthalimido)-4[2-(phenyl)-ethen-1-yl]azetidinone; cis-1-phenoxycarbonyl-3-(2phthalimido)-4-phenyl-azetidinone; 1-phenoxycarbonyl-3-[5-phenyl-oxazolidin-2-on-3-yl]-4-[2-(3-pyridyl)-ethen-1-yl]azetidinone; 1-phenoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)-ethen-1-yl]azetidinone; 1 -phenoxycarbonyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[2-(3-pyridyl)ethen-1-ylazetidinone hydrochloride; 1-phenoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethene-1-yljazetidinone hydrochloride; 1-phenoxycarbonyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl-ethen-1-yljazetidinone hydrochloride; 1-phenoxycarbonyl-3S-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone hydrochloride; 1-phenoxycarbonyl-3R-[4S-phenyl-oxazole idi n-2-on-3-ylJ-4R-[2-(quinoline Jethen-1-yljazetidinone hydrochloride; 1-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yl]azetidinone; 1 -phenoxycarbonyl-3S-[4Sphenyl-oxazolidin-2-on-3-ylJ-4R-[2-(phenyl Jeten-1 -yl] azeti di i η ο n; 1 phenoxycarbonyl-3-benzyl-4-[2-(phenyl)ethen-1 -yl]azetidinone; cis-1 -phenoxycarbonyl-3-benzyl-4-[2-(3-pyridyl)ethen-1 -ylazetide ion hydrochloride; cis-1-phenoxycarbonyl-3-benzyl-4-[2-(cyano)ethen-1-yl]azetidinone; 1 -phenoxycarbonyl-3-(2-phthalimido)-4-buten-1-ylazetidinone; 1 -phenoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-ylazetidinone carboxylic acid; cis-1 -phenoxycarbonyl-3-[oxazolidin-2-on-3-yl]-4-[2-(phenyl)ethen-1-ylazetidinone;

-phenoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-pyridyl)ethen-1-yljazetidinone; trans-1-phenoxycarbonyl-3R-[5-(isopropyl)oxazolidin-2-on-3-yl]-4R-[2-(phenyljeten-1-yljazetidinone; cis-1-phenoxycarbonyl-3-[(5-isopropyl)oxazolidin-2-on-3-yl]-4-[2-(phenyljeten-1-yljazetidinone; 1 -phenoxycarbonyl-3R-[4R-phenyloxazo21

Idin-2-on-3-yl]-4S-[2-(pheni1)ethen-1-yljazetidinone; 1-phenoxycarbonyl I-3 R-[4S-phenyl-oxazolidin-2-one-3-yl IJ-4 R-[2(N-oxide-3-pyridyl)ethen-1-yljazetidinone; trans-1-phenoxycarbonyl-3-benzyl-4-[2-(phenyl)ethen-1-yl]azetidinone; 1-Phenoxycarbonyl-3R-[4S-phenyloxazolide in-2-on-3-yl]-4R-[2-(pyridin-4-yl) Iethen-1-yljazetidinone hydrochloride; 1-[(4-chlorobutyl)oxycarbonyl]-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yljazetidinone; 1 -phenoxycarbonyl-3R-[4R-phenyl-oxazolidin-2-on-3-ylJ-4S-[2-(phenyl-jeten-1-yljazetidinone; cis1-phenoxycarbonyl-3(2-phthalimido)-4-buten-1-yl-azetidinone; trans1-phenoxycarbonyl-3-[4S-phenyl-oxazolidin-2-on-3-ylJ-4-[2-(naphthyl I) ethene-1 -ylazetidinone; trans-1-phenoxycarbonyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[2-(naphthyl)ethen-1-ylazetidinone; trans-1-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3methoxyphenyl)ethen-1-yl]azetidinone; trans-1-phenoxycarbonyl-3R[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-nitrophenyl)ethen-1-ylazetidinone; trans-1-phenoxycarbonyl-3R-14S-phenyloxazolidin-2-on-3-yl]-4R-ethen-1-ylazetidinone; trans-1-phenoxycarbonyl-3R-14phenyl-oxazolidin-2-on-3-yl]-4R-[2-(2-thiophenyl)ethen-1-yl]azetidinone; trans-1-phenoxycarbonyl-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2 (3-furanil)ethen-1-yl]azetidinone; trans-1-phenoxycarbonyl-3R-14-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(5-pyrimidinyl)ethen-1-yl]azetidinone; trans-1-phenoxycarbonyl-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3,5-dimethylphenyl)ethen-1-yl]azetidinone; trans-1 -phenoxycarbonyl-3R-[4-phenyloxazolidin-2-on-3-yl]-4R-[2-(2-nitrophenyl)ethen-1-yljazetidinone; trans-1-phenoxycarbonyl-3R-f4-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(4-nitro-phenyl I ethene-1-yljazetidinone; trans-1-phenoxycarbonyl-3 R-[4-phenyloxazo I idin-2-on-3-yl-4R42(2-nitro-naphthyl)ethen-1-yljazetidinone] nil-3R[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2(4-nitronaphthy l Jethen-1-yljazetidinone; trans-1-phenoxycarbonyl-3R-f4-phenyloxazolid in-2-on-3-ylJ4R-[2-(2-quinolinyl)-ethen-1-yl]azetidinone; trans-1-phenoxycarbonyl22

R-[4-phenyl-oxazolidin-2-oη-3-i I]-4 R-[2-(4-kyno I i n i I )ethéη-1 -i I]azetidinone; trans-1-phenoxycarbonyl-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(7-quinolinyl)ethen-1-yl]azetidinone; trans-1-phenoxycarbonyl-3R-[4-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-isoquinolinyl)ethen-1-yl]azetidinone; trans-1-phenoxycarbonyl-3R-r4-phenyloxazolidin-2-on-3-yl]-4R-[2(8-isoquinolinyl)ethen-1-yl]azetidinone;

trans-1-phenoxycarbonyl-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2(2-benzothiazolyl)ethen-1-yl]azetidinone; and trans-1-phenoxycarbonyl-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2(2-benzoxazolyl)ethen-1-yl]azetidinone; 1-isopropyloxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone; trans-1-sec-butoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone; trans-1-p-nitrobenzyloxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-ylazetidinone; trans-1[2-(2-isopropyl-5-methyl)cyclohexyl]oxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone; trans-1-methoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone; trans-1-benzyloxycarbonyl-3R-14S-phenyl-[oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone;

trans-1-cyclohexyloxycarbonyl-3R-14S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone; trans-1-cyclobutoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen1-yl]azetidinone; trans-1-cycloDentoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone; trans-1 -a 11 i I -oxy - carbo n i I-3 R -[4S-phen i l-oxazolidin-2-on-3-i I]-4 R -[2-(3-pyridyl)ethen-1 -yl]azetidinone; trans-1-isobutoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone;

trans-1-vinyloxycarbonyl-3R-14S-phenyloxazolidin-2-one-3-ylII-4R-[2(3-pyridyl)ethen-1-yl]azetidinone; trans-1 -14-trifluoromethyl-2-pyrimidyl]-3R-[4S-phenyl-oxazolidin-2-on3-yl]-4R-[2-(3-pyridyl)ethene23

-i l]azetid i non; trans-1-[6-nitro-pyridin-2-yl]-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl Jeten-1-yljazetidinone; trans-1[2-phenylquinazol-4-yl]-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl Jeten-1-yljazetidinone; trans-1 -[benzoxazol-2-yl-3R-[4Sphenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethene-1-yl]azetidine; in-2-on3-ylJ-4R-[2-(3-pyrid i I Jetene-1-yljazetidinone; trans-1-benzylthioamido-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3pyridyl)ethene-1-yl]azetidinone; trans-1-[2-bromophenylthioamidol-3R-[4S-phenyloxazolidin-2-on-3-ylJ-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone; trans-1-f3-fluoro-phenyl-thioamidol-3R-F4S-phenyl-oxazolidin-2-o n-3-yl]4 R-[2-(3-pyrid i I )ethé n-1 - i I] a zeti di η ο n; trans-12-fluorophenylthioamidoJ-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethene-1 -ylazetidinone; trans-1-M-fluoro-phenyl-thioamidoloxycarbonyl-3R-(4S-phenyl-oxazolidin-2-on-3-yl)-4R-[2-(3-pyridyl)ethen1-ylazetidinone; trans-1-Γ2.3.5.6-tetrafluoro-phenyl-thioamidole-3R(4S-phenyl-oxazolidin-2-on-3-yl)-4R-[2-(3-pyridyl)ethen-1-ylazetidinone; trans-1-f3-furfuryl-thioamido-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-ylazetidinone; trans-1-naphthyl-thioamido-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-ylazetidinone; trans-1 -[4-trifluoromethylphenyl-thiolamindol-3R-14Sphenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl-Yethen-1-yljazetidinone).

The compounds of the present invention may be prepared by chemical methods known per se, as well as by further methods described below. The following Reaction Scheme I illustrates general methods for preparing the parent compound of Examples 1-46 of Formula I. In all Reaction Schemes and Examples, unless otherwise indicated, the racemic mixtures of the invention are illustrated by preparing a single isomer.

According to Scheme I, the preferred starting material is an aldehyde of formula (i) in which the group R ² is as defined above in Formula I, and p-anisidine of formula (ii). The reactive groups in the group R ² can be protected by methods known per se. Aldehydes that are not commercially available can be prepared by methods known per se, for example, as described in Method A below. The reaction of the aldehyde of formula (i) with the p-anisidine of formula (ii) produces the imine of formula (iii) [Step 1]. In a separate step, oxalyl chloride of formula (v) and acid of formula (iv) are reacted to produce the acid chloride of formula (vi) in which the group R ¹ is as defined above in Formula I [Step 2]. The reactive groups in the group R ¹ can be protected by methods known per se. The 1-(4-methoxy)phenylazetidinones of formula (vii) can be prepared by the 2+2 cycloaddition of the appropriately substituted reactive imine of formula (iii) and the acid chloride of formula (vi) [Step 3]. The preparation of the appropriate imines of formula (iii) and most of the necessary acid chlorides of formula (vi) is described in U.S. Patent Nos. 4,665,171 and 4,751,299, which are incorporated herein by reference. Those not described are commercially available or can be prepared by methods analogous to those described or by methods known per se.

The next step depends on whether a trans or cis compound is desired. In the case of trans compounds, the 1-(methoxy)phenylazetidinone derivatives of general formula (vii) are epimerized at the 3-position with Li-tert-butoxide to obtain the compounds in the trans configuration [4A. Reaction step]. If the compound subjected to epimerization is a racemic mixture, then racemic trans products are obtained. For example, if a cis compound is epimerized in which the group R ¹ of general formula is phthalimido, the resulting trans mixture consists of compounds in the 3(R), 4(R) and 3(S), 4(S) configurations. If a chiral (optically active) auxiliary is used, the corresponding trans enantiomer is obtained. The epimerized compound (viii) is then oxidized with CAN (ceric ammonium sulfate) to remove the jD-methoxyphenyl substituent [Step 5A]. The resulting compound (ix) is then acylated with the corresponding chloroformate (x) to give the trans-configured oxycarbonyl azetidinone derivative (xi) [Step 6A].

In ^the case of cis compounds, in another way, the 2-methoxyphenyl substituent is first removed from the 1-(methoxy)phenylazetidinone compound (vii) by oxidation with CAN reagent [Step 4B]. The resulting compound (xii) is then acylated with the appropriate chloroformate (x) to give the corresponding substituted oxycarbonylazetidinone derivative (xiii) in the cis configuration.

As mentioned above, the compounds prepared according to Reaction Scheme I of the preparation may be pure diastereoisomers, mixtures of diastereoisomers, or racemates. The exact stereochemical composition of the compound is determined by the given reaction conditions, the combination of substituent groups, and the stereochemical characteristics of the reagents used in Reaction Scheme I. It will be apparent to those skilled in the art that individual diastereoisomers can be recovered from diastereoisomeric mixtures by chromatography or fractional crystallization, if necessary.

Bases useful in Preparation Scheme I include, but are not limited to, lithium bis(trimethylsilyl)amide, aliphatic tertiary amines such as trimethylamine and triethylamine (TEA), dimethylaminopyridine (DMAP), cyclic tertiary amines such as N-methylpiperidine and N-methylmorpholine, aromatic amines such as pyridine and lutidine, and other organic bases such as 1,8-diazabicyclo[5.4.0]undec-7-ene (DBU).

Solvents useful in Preparation Scheme I include those which dissolve the reactants and do not interfere with the reaction. Solvents useful in Preparation Scheme I include, but are not limited to, dioxane, ethyl acetate, acetonitrile (CH ₃ CN), dimethyl sulfoxide, N,N-dimethylformamide, tetrahydrofuran (THF), ethyl formate, N,N-dimethylacetamide, hexane, diethyl ether, benzene, toluene, tert-butyl methyl ether, diisopropyl ether, ethylene glycol dimethyl ether, dichloromethane, chloroform, carbon tetrachloride, trichloroethylene, and 1,2-dichloroethane, either alone or as a mixed solvent.

To prepare acid addition salts of the compounds of the present invention, the above-mentioned acids are also added to the product after acylation.

The primary amine, the substituent or protecting group of the primary amine, the solvent used in the reaction, and the reaction temperature, among other parameters, influence whether the formation of one or the other isomer is more favorable.

The following is a more detailed description of the reaction steps described in Preparation Scheme I. Although the chemical methods described below can be used to prepare azetidinone derivatives bearing substituents represented by the general formulae R ¹ and R ² , for the sake of simplicity ^, the group represented by the general formula R ¹ is depicted as a phenyl oxazolidinone (or phthalimido) group, the group represented by the general formula R ² is depicted as a (2-phenyl)ethen-1-yl and (2-pyridin)ethen-1-yl group, and the group represented by the general formula R 3 is depicted as a phenyloxycarbonyl group. Those skilled in the art will recognize that the different groups represented by the general formula R can be replaced by other groups, if appropriate, without changing the overall chemical behavior of the compounds. The following descriptions and examples further illustrate the preparation of the compounds of the present invention, but are for illustrative purposes only and are not intended to limit the scope of the invention in any way. Other methods of preparing the compounds of the present invention, even if not illustrated, are considered to be within the scope of the present invention.

A. METHOD: Preparation of Aldehydes of Formula I As indicated above, most of the aldehydes used are commercially available. Aldehydes which are not commercially available can be prepared, for example, by methods known per se, such as oxidation of primary alcohols in the presence of pyridinium chlorochromate or reduction of acid chlorides with lithium tert-butoxyaluminum hydride at -78°C. The aldehydes are preferably prepared by the method described in Anqew. Chem., 87, 486 (1975), i.e., one equivalent of the appropriately substituted carboxyaldehyde is reacted with one equivalent of FMTTP [formylmethylenetriphenylphosphorane, (Ph) ₃ PCHCHO] in the presence of a solvent, which is preferably benzene.

or toluene. The reaction mixture is stirred at a temperature of about 75°C to about 85°C, preferably about 80°C, until all of the starting material has reacted as indicated by the NMR spectrum. If excess starting material is present, additional forml-methylenetriphenylphosphorane can be added to the reaction mixture, which reacts with the excess carboxyaldehyde. The final product can be recovered by conventional methods, such as filtration and/or removal of the organic solvents under reduced pressure.

In more detail, for example, aldehydes are prepared by the method according to Reaction (A):

Objective: Production of aldehyde on a large scale Procedure:

Materials Molecular mass Quantity Moth Equivalent pyridine 107.11 100g 0.9336 1.0 FMTPP 304.33 284.13 grams 0.9336 1.0

100 g of 3-pyridinecarboxaldehyde, 6 L of benzene (solvent), and 284.13 g of formylmethylenetriphenylphosphorane (FMTPP) are charged into 1-liter round-bottom flasks. The resulting suspension is heated to 80°C and stirred at this temperature overnight. The completion of the reaction is monitored by NMR. The NMR spectrum shows the presence of starting material. Another 20% portion of FMTPP is added to the reaction mixture and stirred overnight. At this time, the NMR analysis shows only a very small amount of starting material. The reaction mixture is cooled to room temperature and the organic solvents are removed under reduced pressure. 3 L of ice-cold diethyl ether are added to the residue and stirred. The solids are filtered off and the solvent is removed under reduced pressure. The residue (123 g) is obtained as a dark solid. Yield: 99.8%. MS = 132 «««<···· * · u,

REACTION STEP 1: Preparation of imines of formula (iii) The imines of the present invention are prepared by methods known per se, for example by charging one equivalent of the appropriate aldehyde to a flask with a solvent, which is advantageously dichloromethane. To the mixture is then added one equivalent of the primary amine (£>-anisidine). A drying agent, typically magnesium sulfate, sodium sulfate or silica gel, is added to the reaction mixture in an amount of approximately 1 g for every 3.8 g of amine. The reaction mixture is stirred overnight at room temperature until the reactants are completely consumed, as measured by any available method. The mixture is then filtered and the solvent is removed under reduced pressure to give the desired imine.

In more detail, for example, imines are prepared by the method according to Reaction (B):

Objective: Large-scale production of imine.

Proceedings:

Materials Molecular mass Quantity Moth Equivalent aldehyde 132.1 120g 0.9311 1.0 anisidine 123.2 114.7g 0.9311 1.0

A 1-liter round bottom flask was charged with approximately 120 g of aldehyde and 1 L of dichloromethane. To this mixture was added 114.7 g of ρ,-anisidine and 0.8 L of dichloromethane. Thirty (30) g of magnesium sulfate was then added as a drying agent. The reaction mixture was stirred overnight at room temperature. NMR indicated that the reaction was complete. The magnesium sulfate was removed by filtration and the solvent was removed under reduced pressure to give 210.65 g of imine. Yield = 94.98%. MS = 238

2. Reaction step: Preparation of acid chloride of general formula (vi)

The acid chlorides used in the present invention are also prepared by methods known per se, such as the process described in U.S. Patent Application No. 4,665,171, which is incorporated herein by reference. For example, to prepare the acid chloride, 1.8 molar equivalents of oxalyl chloride are reacted with one equivalent of the appropriately substituted acid in the presence of a solvent (preferably toluene or benzene). The reaction mixture is heated to about 60°C for about 3-4 hours under an inert gas blanket (i.e., nitrogen gas, helium gas, or argon gas). The reaction mixture is cooled to room temperature, and the organic solvent is removed under reduced pressure.

For example, acid chlorides are prepared according to Reaction (C).

Substances Molar mass Quantity mol Equivalent acid 132.1 120 g 0.9311 1.0 oxalyl chloride 123.2 114.7 g 0.9311 1.0

The acid was suspended in 2840 ml of toluene at room temperature. 103.6 g of oxalyl chloride was added in one portion. The solution was heated to 60°C for 4 hours while purging with nitrogen. The solution was then cooled to room temperature and the toluene was removed under reduced pressure to give an oil. The oil solidified on standing below 0°C, so

113 g of acid chloride are obtained.

REACTION STEP 3: 2+2 Cycloaddition

The 2 + 2 cycloaddition is a well-known process. A detailed description of the process can be found in G. I. Georg, Ed.; Organic Chemistry of β-Lactams, Verlag Chemie, New York, 1992

6. and in U.S. Patent Applications Nos. 4,751,299 and 4,665,171, which are incorporated herein by reference. In general, 1 equivalent of the appropriate acid chloride and dichloromethane are charged to a flask and the mixture is cooled to about -78°C. 1.5 equivalents of a tri(C1-4 alkyl)amine, typically triethylamine, are then added to the mixture, taking care not to exceed 70°C. The reaction mixture is stirred thoroughly. The imine dissolved in dichloromethane (10 mL dichloromethane/1 g imine) is slowly added while maintaining the temperature near -70°C. Although the time of addition is not critical, the best results are generally obtained when the imine is added dropwise over a period of about 1 to about 6 hours. The reaction mixture is then stirred for an additional period of about 2 to about 4 hours. The reaction is carried out at a temperature of about -78°C to about 25°C, preferably about -78°C to about 0°C, more preferably about 78°C to -75°C in a solvent in the presence of a tri(C1-4 alkyl)amine. The reaction mixture is then allowed to warm to room temperature and the reaction is quenched by the addition of a saturated aqueous ammonium chloride solution. The resulting mixture is separated and the solvent is removed under reduced pressure. The solid residue is stirred with ethyl acetate for 1 hour. The resulting wet suspension is washed with a large amount of ethyl acetate. Finally, the material is washed with ether. The resulting product was dried overnight under reduced pressure at 40°C.

In more detail, for example, the 2+2 cycloaddition is carried out according to the method of Reaction (D):

Objective: Large-scale production of β-lactam by 2+2 cycloaddition

Proceedings:

Materials Molecular mass Quantity mole Equivalent imin 238 210.65 grams 0.8851 1.0 acid chloride 239 210.9g 0.8825 1.0 TEA 101.2 133.96 grams 1,324 1.5

A 1-liter round-bottomed flask is charged with 210.9 g of acid chloride and 2 L of dichloromethane. The mixture is cooled to -78°C. To this pink/purple mixture is added 133.96 g of triethylamine, keeping the temperature near 70°C. The reaction mixture is stirred for 15 minutes. The imine dissolved in 2 L of dichloromethane is added slowly, keeping the temperature near -70°C. The addition time is approximately 5 hours. The reaction mixture is stirred for 2 hours at -75°C. The reaction mixture is then slowly warmed to room temperature. Thin layer chromatography (TLC) is performed at 0°C. The TLC performed at room temperature does not differ from the results of the study performed at 0°C. The reaction was quenched by adding 3 L of saturated ammonium chloride solution (about 500 g NH ₄ Cl), the product was separated, and the organic solvent was removed under reduced pressure. To the solid residue was added 1 L of ethyl acetate and stirred for 1 hour, the resulting wet material was washed with a large amount of ethyl acetate. The material was finally washed with diethyl ether. The resulting product was dried at 40°C under reduced pressure overnight. Total weight = 176 g. Yield = 46%. MS = 441.3

Analysis

Calcd (%): C 70.73 H 5.25 N 9.52

Found (%): C 70.60 H 5.14 N 9.43

REACTION STEPS 4A - 6A: Preparation of TRANS isomers REACTION STEP 4A: Epimerization

The product of the 2+2 cycloaddition reaction is epimerized with lithium tert-butoxide to give the trans-configuration isomers. Surprisingly, we discovered that when the epimerization is carried out with lithium tert-butoxide, higher yields and higher purity products are obtained than when other reagents, such as potassium tert-butoxide, are used.

The epimerization of the substituted cis azetidinone to the substituted trans azetidinone is easily accomplished by adding 1.1 equivalents of lithium tert-butoxide to one equivalent of the material [the substituted (methoxy)phenyl azetidinone dissolved in a solvent, preferably dry tetrahydrofuran, at about 0°C. The reaction is quenched with saturated ammonium chloride solution and the reaction mixture partitioned. The organic layer is treated with activated carbon (Darco™), dried over magnesium sulfate, filtered, and then dried under reduced pressure.

In more detail, for example, epimerization is carried out by the method according to Reaction (E):

Objective: Epimerization of the product of the 2+2 cycloaddition

with lithium tert-butoxide Procedure: Substances Molar mass Quantity mole Equivalent cis 2+2 441 50.0 g 0.1134 1.0 Li-EOBu 80.05 9.98 g 0.1247 1.0

50.0 g of 1-methoxyphenyl-3S-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (cis 2+2) and 2500 ml of dry tetrahydrofuran are charged into a 1-liter round-bottomed flask at room temperature.

The mixture was cooled to 0°C in an ice bath. 9.98 g of lithium tert-butoxide was added in one portion. The reaction mixture was stirred for approximately 20 minutes. The ice bath was removed and the mixture was allowed to warm slowly to room temperature. The intermediate product that had come out of solution redissolved at 2°C. The mixture was found to contain a small amount of cis material by NMR spectroscopy at room temperature. The mixture was stirred overnight at room temperature. The NMR spectroscopy indicated that it still contained a small amount of cis isomer. The reaction was quenched by adding 2 L of saturated ammonium chloride solution. The organic layer was treated with Darco™, dried over magnesium sulfate and filtered. The solvent was removed under reduced pressure. Total weight = 42.7 g. Yield = 85.4%. MS = 441 Analysis Calculated (%); C 70.74 H 5.25 N 9.52

Found (%): C 70.96 H 5.39 N 9.55

REACTION STEP 5A: Oxidation with cerium ammonium nitrate

Oxidation with cerium ammonium nitrate (CAN) is known, see for example J. Orq. Chem., 47, 2765-68 (1982). When a compound is reacted with CAN in the presence of acetonitrile under relatively mild conditions, the methoxyphenyl group is removed. In more detail, for example, the oxidation of the trans isomer can be carried out according to the following, Reaction (F):

t

Objective: Oxidation with cerium ammonium nitrate Procedure:

Materials Molecular mass Quantity mole Equivalent public production 441 35.78 grams 0.0811 1.0 CAN 548.2 177.9g 0.3245 1.0

A 1-liter flask was charged with 35.78 g of 1-(4-methoxyphenyl)-3R-[4phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (intermediate term) and 3.5 L of acetonitrile solvent. The mixture was cooled to -10°C, and at this temperature 177.9 g of cerium ammonium nitrate and 3.5 L of deionized water were added. The course of the reaction was monitored by thin-layer chromatography with ethyl acetate. Since the reaction was not complete, another 0.5 equivalent of CAN reagent was added to the mixture and stirred for 1 hour. At this time, the reaction was complete according to TLC. To the reaction mixture was added 7.0 L of sodium bicarbonate solution and 3.0 L of ethyl acetate, and the mixture was warmed to room temperature and filtered through a pad of Celite™. The layers were separated and the aqueous layer was extracted twice with 2 L of ethyl acetate. The combined organic layers were washed with 4 L of 10% sodium sulfite solution. The organic layer was then treated with Darco™ and dried over magnesium sulfate. The organic solvent was removed under reduced pressure. Total weight = 169 g. Yield = 62.36%. MS = 335.2 Analysis Calcd (%); C 68.05 H 5.11 N 12.53

Found (%): C 68.06 H 5.11 N 12.43

REACTION STEP 6A: Acylation

The acylation is carried out by dissolving the substituted azetidinone, preferably in dichloromethane. A base is then added to the reaction mixture, followed by the addition of the substituted chloroformate, and the reaction mixture is stirred until the reaction is complete. The reaction is then quenched with ammonium chloride, treated with Darco™, and dried on a desiccant. The resulting solid is then washed with solvent and dried. In more detail, for example, the acylation is generally carried out according to the method of Reaction (G).

Objective: Acylation of free amine. Equivalent Procedure: Quantity mole Materials Molar mass public production 334.0 15.0g 0.0449 1.0 Materials Molar mass Quantity mole Equivalent DMAP 122.2 catalytic TEA 101.2 9.09 ml 0.0898 2.0 PCF 156.6 14.06 ml 0.0898 2.0

A 1-liter round-bottomed flask was charged with 15.0 g of 3R-[4S-phenyloxazolidin-2-one 3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (intermediate) and 4690 ml of dichloromethane. Dimethylaminopyridine (DMAP) was added at room temperature and stirred for 10 minutes. TEA was added in one portion and the mixture was stirred for 10 minutes. Then phenyl chloroformate (PCF) was slowly added. After the addition was complete, TLC showed that the starting material remained in the mixture. The mixture was stirred at room temperature for approximately 2 hours. TLC showed that the ratio of the components had not changed. An additional 22 equivalents of TEA and PCF were added to the reaction mixture and stirred for 2 hours. TLC showed that there was no change. After adding another 2 equivalents of TEA and PCF and stirring for 1.5 hours, the TLC result showed little change. At this time, 4.0 equivalents of TEA and PCF and 0.1 equivalent of DMAP were added. The mixture was stirred at room temperature overnight. According to TLC, the ratio of the components had changed little. The reaction was quenched by adding 4 L of saturated ammonium chloride solution, the mixture was treated with Darco™ and dried over magnesium sulfate. After removing the solvent, a white solid was obtained. The solid was suspended in diethyl ether and filtered. The solid was insoluble in 4 L of ethyl acetate, so 4 L of dichloromethane was added to dissolve it. 72 mL of 1M ethereal hydrochloric acid was added to the mixture and stirred for one hour. Removal of the organic solvent under reduced pressure gave a white solid. The solid was suspended in ether and filtered. The product was dried at 60°C for a long time to remove residual solvents. Total weight - 6.2 g. Yield = 30.4%. MS = 455.5 Analysis

Calculated (%): C 63.48 H 4.51 N 8.54 Cl 7.21

Found (%): C 63.27 H 4.46 N 8.44 Cl 6.97

4B. - 5B- REACTION STEP: Preparation of cis isomers

The cis isomeric compounds of the present invention are prepared by acylating the product of the above 2+2 cycloaddition reaction after oxidation with CAN as follows.

REACTION STEP 4B: Oxidation with Cerium Ammonium Nitrate (CAN)

Oxidation with cerium ammonium nitrate (CAN) is known, see, for example, J. Orq. Chem., 47, 2765-68 (1982). The procedure is carried out in the same manner as above, in reaction step 5A. In more detail, for example, the oxidation can be carried out according to the following, Reaction (H):

Objective: Oxidation with cerium ammonium nitrate

Proceedings:

Materials Molecular mass Quantity mole Equivalent public production 441 35.78 grams 0.0811 1.0 CAN 548.2 177.9g 0.3245 1.0

The cis-1-(4-methoxyphenyl)-3S-(2-phthalimido)-4R-F2-(phenyl)ethen-1-ylazetidinone (intermediate term) was suspended in 600 ml of acetonitrile and cooled to 0°C. A solution of CAN in 600 ml of water was then added dropwise to the cloudy reaction mixture at 0°C. The mixture was left to stand covered for 30 minutes, then stirred at 0°C for a further 30 minutes and extracted with 500 ml of ethyl acetate. The aqueous phase was again extracted twice with 300 ml of ethyl acetate. The organic phase was then extracted with 1 x 500 ml of 5% sodium bicarbonate solution and then 3 x 500 ml of 10% sodium sulfite solution. The combined organics were suspended in 50 g of Darco™ for 30 min, then 100 g of magnesium sulfate was added. The mixture was stirred for an additional 15 min and then filtered through a pad of Celite. The solution was concentrated under reduced pressure to give an oil. The oil was dissolved in 60 mL of ethyl acetate and triturated with 250 mL of ether. The mixture was filtered to collect 2.70 g of pure product as a finely divided white solid. The filtrates were further purified by silica gel chromatography.

4.04 g of pure product is obtained. Total weight = 6.74 g. Yield =

44.99%. MS = 317.5

REACTION STEP 5B: Acylation

The acylation is carried out in the same manner as described in Reaction Step 6A. In more detail, for example, the acylation is carried out according to Reaction (11):

Objective: Acylation of free amine.

Proceedings:

Materials Molecular mass Quantity mole Equivalent public production 318 0.2g 0.629 1.0 PCF 156.6 0.16 ml 1.26 mmol 2 LHMDS 0.63ml 0.629 mmol 1.0 The 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone

(intermediate term.) 6 ml of dry tetrahydrofuran were added. The mixture was cooled to -75°C. 1.0 mol of lithium bis(trimethylsilyl)amide (LHMDS) was added dropwise from a syringe under a nitrogen blanket. The mixture was stirred at -75°C for 30 min. PCF was added in one portion and the mixture was stirred at room temperature for 1 h. The reaction was then quenched with ammonium chloride and extracted with ethyl acetate. The organic phase was purified on silica gel. 0.105 g of a white solid was obtained. MS = 438 Analysis Calcd (%): C 71.23 H 4.14 N 6.39

Found (%): C 71.53 H 4.27 N 6.49

Preparation of 1:1 cis:trans isomer mixtures

1:1 mixtures of cis:trans isomers of the compounds of the present invention can be prepared by a combination of steps according to Reaction (I2).

Epimerization and acylation

Materials Moítömeq Quantity mole Equivalent public production 318 2 grams 6.29 mmol 1 PCF 156.6 1.58ml 12.58 mmol 2 TEA 101.2 8.77 ml 62.90 mmol 10 DMAP 122.2 catalytic

The cis intermediate was dissolved in 100 ml of dichloromethane at room temperature. TEA was added in one portion. A solution of phenyl chloroformate in 20 ml of dichloromethane was added slowly dropwise at room temperature. The solution was stirred overnight at room temperature. The reaction was quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel using a 1:1 hexane/ethyl acetate solvent mixture. 1.69 g of a cis:trans isomer mixture was obtained. The mixture was dissolved in 100 ml of ethyl acetate, 300 ml of hexane was added, and crystals of the pure trans isomer were seeded. The mixture was allowed to stand overnight, then filtered and the finely divided white crystals were collected. 99.4% trans, 0.6% cis. The product obtained was 0.89 g of trans isomer. MS = 438 Analysis

Calcd (%): C 71.23 H 4.14 N 6.39

Found (%): C 71.46 H 4.35 N 6.18

The following examples further illustrate the compounds of the present invention and the methods for their preparation. The examples are not intended to limit the scope of the invention and should not be used for that purpose.

The starting materials for Examples 1-46 were prepared according to the general procedures outlined in Preparation Scheme I.

Example 1: trans-1-[4-(fluoro)phenoxycarbonyl)-3-(2-phthalimido)-4-[2(phenyl)ethen-1-yl]azetidinone [Compound of Formula P1]

Materials Molecular mass Quantity mole Equivalence Intermediate production 318 0.150g 0.4716 mmol 1 4-FPCF 174.56 0.124ml 0.9432 mmol 2 TEA 101.2 0.651 ml 4.716 mmol 10 DMAP 122.2 catalytic The 3-(2-phthalimidc >)-4-[2(phenyl)-ethen-1-ylazetidinone public-

The inner product was dissolved in 10 ml of dichloromethane at room temperature. DMAP (4-dimethylaminopyridine) and TEA were added in one portion. 4-fluorophenyl chloroformate (4-FPCF) dissolved in 10 ml of dichloromethane was added dropwise to the reaction mixture at room temperature under a nitrogen blanket. The solution was stirred at room temperature for 24 hours. The reaction was then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 106 mg of a white solid was obtained in 49.3% yield. MS=456

Example 2: trans-1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P2]

Materials Molar mass Quantity mo !_ Equivalent Intermediate production 318 2 grams 6.29 mmol 1 PCF 156.6 1.58ml 12.58 mmol 2 TEA 101.2 8.77 ml 62.90 mmol 10 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone was mixed with 50 ml of dichloromethane at room temperature. TEA and DMAP were added to the mixture in one portion. Phenyl chloroformate (PCF) dissolved in 20 ml of dichloromethane was slowly added dropwise to the reaction mixture at room temperature. The solution was stirred overnight at room temperature. The reaction was then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel with 100% hexane (1:1: hexane/ethyl acetate) as solvent. The resulting product (1.69 g) was a mixture of cis/trans isomers. The mixture is then dissolved in 100 ml of ethyl acetate, 300 ml of hexane are added, and the mixture is seeded with crystals of the pure trans isomer and left to stand overnight. The finely divided, white crystals that precipitate are collected by filtration. The material does not contain any cis isomer according to the ¹ H-NMR spectrum. Its composition by HPLC analysis is 99.337% trans and 0.553% cis isomer. The weight of the obtained trans isomer is 0.89 g. MS=438 Analysis Calculated (%): C 71.23 H 4.14 N 6.39

Found (%): C 71.47 H 4.35 N 6.18

Example 3: trans-1-[4-(nitro)benzyloxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P3]

Materials Molecular mass Quantity mole Equivalent Intermediate production 318 0.1g 0.3144 mmol 1 4-nitro 215.6 0.136 g 0.6288 mmol 2 TEA 101.2 0.438 ml 3.144 mmol 10 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2(phenyl)-ethen-1-yl]azetidinone was dissolved in 4 mL of dichloromethane at room temperature under a drying agent. TEA and DMAP were added in one portion. Then, a solution of 4-nitrobenzyl chloroformate (4-nitro) in 6 mL of dichloromethane was added dropwise at room temperature. The resulting mixture was stirred overnight and the reaction was quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 123 mg of a white solid was obtained in 79% yield. MS=497

4, Example

-[(4-chlorobutyl)oxycarbonyl]-3-(2-phthalimido)-4-[2(phenyl)ethene-1-

il]azetidinone [Compound with formula P4] Materials Molecular mass Quantity mole Equivalent public production 318 0.5g 1.57 mmol 1 TEA 101.1 2.1 8m1 15.70 mmol 10 4C 171.02 0.43m1 3.14 mmol 2 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2(phenyl)ethen-1-yl]azetidinone was dissolved in 10 mL of dichloromethane at room temperature. TEA and DMAP were added in one portion, followed by the dropwise addition of 4-chlorobutyl chloroformate (4C) in 10 mL of dichloromethane at 25°C. The reaction mixture was stirred at room temperature using a tube containing a drying agent for 72 h. The reaction was then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 204 mg of a 1:1 cis/trans isomer mixture was obtained. Yield = 57%. MS = 451.9

Example 5: trans-1-[(4-chlorobutyl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P5]

The trans isomer is obtained by the procedure of Example 4. The trans isomer is separated from the silica gel instead of the cis isomer. Yield = 57%. MS = 451.2

Example 6: trans-1-[(4-iodobutyl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P6]

Substances Molar mass Quantity mol Equivalent value 452 0.1 g 0.2212 mmol 1

Nal 150 0.166 g 1.106 mmol 5

The product obtained in Example 5 and sodium iodide were dissolved in 10 ml of acetone. The mixture was stirred overnight at room temperature and then filtered. The filtrate was concentrated under reduced pressure to give 73 mg of a yellow solid. Yield = 60.6%. MS = 544

Example 7

-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yljazetidinone

[Compound with formula P7] Quantity mole Equivalence Materials Molar mass Intermediate production 318 0.2 0.629 mmol 1 PCF 150.6 0.16 ml 1.26 mmol 2 TEA 101.2 0.88ml 6.19 mmol 10 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone was mixed with 5 mL of dichloromethane at room temperature. TEA and DMAP were added to the mixture in one portion. Phenyl chloroformate (PCF) was added dropwise. The solution was stirred at room temperature for 16 h. The reaction was then quenched with ammonium chloride solution and then ethyl acetate was added.

The organic phase was purified on silica gel to give 120 mg of a white solid as a 1:1 cis/trans isomeric mixture. Yield = 43.5 MS=438

Example 8: trans-1-benzyloxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)ethen-1yl]azetidinone [Compound of Formula P8]

Materials Molar mass Quantity mole Equivalent Intermediate production 318 0.150g 0.4716 mmol 1 BCF 110.6 0.135m1 0.9432 mmol 2 TEA 101.2 0.657m1 4.716 mmol 10 DMAP 122.2 catalytic The 3S-(2-ftá Ii m time) +4-[2-(phenyl)ethen-1-yl]azetidinone public-

The product was mixed with 5 ml of dichloromethane at room temperature. TEA and DMAP were added in one portion at room temperature, followed by dropwise addition of benzyl chloroformate (BCF). The reaction mixture was stirred at room temperature for 24 h, then the reaction was quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 160 mg of a white solid was obtained. Yield = 75% MS = 452.2 Analysis Calcd (%): C 71.67 H 4.45 N 6.19

Found (%): C 71.86 H 4.95 N 6.17

Example 9

-[(2-propen-1-yl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of formula P9]

Materials Molecular mass Quantity mole Equivalence Intermediate production 318 0.2g 0.6289 mmol 1 AC 120.54 0.13 ml 1.2578 mmol 2 TEA 101.2 0.88ml 6.289 mmol 10 □ MAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone was mixed with approximately 8 mL of dichloromethane at room temperature. TEA and DMAP were added in one portion. Allyl chloroformate (AC) was added dropwise at room temperature. The reaction mixture was stirred at room temperature overnight and quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 185 mg of a white solid was obtained. Yield = 73% MS = 402.4

Example 10

-[(2-isopropyl-5-methyl)cyclohexyl]oxycarbonyl-3-(2-phthalimido)-4-[2(phenyl)ethen-1-yl]azetide [Compound of formula P10]

Mothers Molecular mass Quantity mole Equivalent Intermediate production 315 0.2g 0.629 mmol 1 5MC 219 0.28g 1.26 mmol 2 TEA 101.2 0.88ml 6.29 mmol 10 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone was dissolved in 5 mL of dichloromethane at room temperature. TEA and DMAP were added in one portion, followed by 2-menthyl chloroformate (5MC) dropwise. The reaction mixture was stirred at room temperature for 24 h, then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 105 mg of a white solid was obtained. Yield = 33.4% MS=500

Example 11: trans-1-[4-(methoxycarbonyl)phenoxycarbonyl]-3-(2-phthalimido)-4[2(phenyl)ethen-1-yl]azetidinone [Compound of Formula P11]

Materials Molecular mass Quantity mole Equivalent Intermediate production 318 1g 3.14 mmol 1 TEA 101.2 4.38m1 31.45 mmol 10 MCPCF 214.6 1.35g 6.28 mmol 2 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1 yl azetidinone was dissolved in 25 mL of dichloromethane at room temperature in a flask sealed with a desiccant tube. TEA and DMAP were added in one portion, then 4-methoxycarbonylphenyl chloroformate (MCPCF·) dissolved in 10 mL of dichloromethane was slowly added dropwise while the reaction mixture was vigorously stirred. The reaction mixture was stirred at room temperature overnight, then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel with 100% hexane (1:1 hexane/ethyl acetate). 750 mg of a white solid was obtained.

Yield = 48.2% MS=496

Example 12: trans-1-[4-(methoxy)phenoxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P12]

Materials Möltömeq Quantity mole Equivalent Int. production TEA P-OCH3PCF DMAP The 3- 318 0.5 g 1.57 mmol 1 101.2 2.19 ml 15.72 mmol 10 186.6 0.467 ml 3.14 mmol 2 122.2 catalytic -(2-phthalimido)-4-[2(phenyl)-ethen-1-yl]azetidinone intermediate- dissolve the internal product at room temperature 25 ml dichloro-

in methane. TEA, DMAP, and then 4-ethoxyphenyl chloroformate (p-OCH ₃ PCF) were added dropwise. The reaction mixture was stirred at room temperature for 16 h, then quenched with 0 N hydrochloric acid. The organic phase was separated and purified on silica gel. 370 mg of a white solid was obtained. Yield = 50.4% MS=468

Analysis

Calcd (%): C 69.23 H 4.30 N 5.98

Found (%): C 69.11 H 4.28 N 5.94

Example 13: trans-1-[(2-propen-1-yl)oxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P13]

Materials Möltömeq Quantity mole Equivalent Intermediate production 318 0.150g 0.4716 mmol 1 chloroform 256.6 0.242g 0.9432 mmol 2 TEA 101.2 0.657 ml 4.716 mmol 10 DMAP 122.2 catalytic The 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone public- indoor product at room temperature in tetrahydrofuran sol-

Add TEA and DMAP in one portion. Add chloroform in tetrahydrofuran dropwise to the reaction mixture at room temperature. Stir overnight and partition between ethyl acetate and 1 N hydrochloric acid. Purify the organic phase on silica gel. 150 mg of a white solid is obtained. Yield = 61% MS = 522

Example 14

-phenoxy-carbonic acid-3-(2-phthalamide)-4-[2-(3-pyridyl)ethene-1-yl]azetidinone [compound with formula P14]

Materials Molar mass Quantity mole Equivalent value Intermediate production 319 100mg 0.3134 mmol 1 TEA 101.2 0.437 ml 3.134 mmol 10 PCF 156.6 0.074 ml 0.6268 mmol 2 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(3-pyridyl)ethen-1-yl]azetidinone was dissolved in dichloromethane at room temperature. TEA and DMAP were added in one portion. A solution of phenylchloroform (PCF) in dichloromethane was added dropwise to the reaction mixture via syringe. The operation was carried out in a flask sealed with a tube containing a drying agent. The reaction was then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 89 mg of a white solid was obtained. Yield = 64.5% MS=440

Example 15

-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(3-pyridyl)ethen-1-yl]azetidinone hydrochloride [Compound of formula P15]

50 mg of the compound obtained in Example 14 was dissolved in 2 ml of ethyl acetate. 0.5 ml of 1 N ethereal hydrochloric acid solution was added. The reaction mixture was stirred at room temperature for 1 hour. The white solid was separated by filtration to give 44 mg of product. Yield = 81.3% MS=440

Example 16: cis-1-phenoxycarbonyl-3-(2-phthalimido)-4-(2-phenyl)ethyl]azetidinone

[Compound with formula P16]

Materials Molar mass Quantity mole Equivalent Intermediate production 316 — 0.4716 mmol 1 PCF 156.6 0.118 ml1 0.9432 mmol 2 TEA 101.2 0.66 ml1 4.716 mmol 10 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4[2-(phenyl)ethyl]azetidinone, phenyl chloroformate (PCP), and DMAP were mixed in 10 mL of dichloromethane at room temperature under a nitrogen blanket. A solution of TEA in 10 mL of dichloromethane was added dropwise under a nitrogen blanket. The reaction mixture was stirred at room temperature until the reaction was complete. The reaction was then quenched with 1 N hydrochloric acid, the organic phase was separated, and the reaction mixture was concentrated in vacuo.

choose and on silica gel we clean. 130mg white solid material is obtained. Yield = Analysis 63% MS =440 Calculated (%): C 70.94 H 4.58 N 6.36 Score (%): C 70.64 H 4.53 N 6.36

Example 17. Trans-1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(carboxy)ethen-1-yl]azetidinone [Compound P17] mg of (±) trans-1-phenoxycarbonyl-3-(2-phthalimido)-4-(2-(carboxybenzyl)ethen-1-yl]azetidinone intermediate was dissolved in 3 ml of dichloromethane at room temperature and 10 mg of 5% Pd on charcoal was added. The reaction mixture was stirred under a hydrogen atmosphere. Thin layer chromatography revealed the appearance of a new spot. The mixture was stirred under a hydrogen atmosphere overnight. The organic phase was separated and purified on silica gel. 25.2 mg of a white solid was obtained. Yield = 62% MS=408

Example 18

-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(diphenyl)ethen-1-yl]-azetidinone [compound of formula P18]

Materials Molar mass Quantity mole Equivalent Intermediate production 394 125mg 0.3172 mmol 1 PCF 156.6 0.08 ml 0.6344 mmol 2 TEA 101.2 0.442 ml 3.172 mmol 10 DMAP 122.2 catalytic

5ml of the intermediate 3-(2-phthalimido)-4-[2-(diphenyl)ethen-1-yl]-azetidinone was dissolved in dichloromethane at room temperature. TEA and DMAP were added in one portion. Phenyl chloroformate (PCF) was added dropwise at room temperature and stirred overnight. The reaction was then quenched with 1N hydrochloric acid. The organic phase was separated and purified on silica gel. 104 mg of a white solid was obtained. Yield = 63.7% MS=514

Example 19: cis-1-[4-(methoxycarbonyl)phenoxycarbonyl]-3-(2-phthalimido)-4-[2(phenyl)ethen-1-yl]azetidinone [Compound of Formula P19]

Materials Molar mass Quantity mole Equivalent Intermediate production 318 0.1g 0.3144 mmol 1 MCP 152 96mg 0.6288 mmol 2 phosgene · 1.93M 0.326 ml 0.6288 mmol 2 pyridine 79.10 0.069 ml 0.6288 mmol 2

4-Methoxycarbonylphenol (MCP) was mixed with dichloromethane at room temperature. A 1.93 M phosgene solution was added dropwise from a syringe under a nitrogen blanket. The solution was cooled to -60°C. Pyridine was added dropwise from a syringe. The reaction mixture was stirred at -60°C for 15 min and then warmed to -10°C for 1 h. The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone was added in one portion from a syringe. No reaction was observed by TLC. At this time, 10 equivalents of RA were added in one portion to the reaction mixture and the reaction mixture was stirred at room temperature overnight. The reaction was then complete by TLC. The reaction was quenched with 1N hydrochloric acid. The organic phase was separated and purified on silica gel. 84 mg of a white solid was obtained. Yield = 54% MS=496

Example 20: cis-1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-ylazetidinone [Compound P20] mg of 3-(2-phthalimido)-4-[2-(phenylethen-1-ylazetidinone intermediate is dissolved in ethanol in a Parr hydrogenation apparatus. Lead-poisoned calcium carbonate containing 5% palladium (Lindlar catalyst, KÁT) is added. The reaction mixture is shaken at room temperature with 20 psig hydrogen pressure in the Parr apparatus for 2 hours. The solid is purified on silica gel. 16 mg of product is obtained. Yield = 79.6% MS=438

Example 21: trans-1-[4-(chloro)phenoxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P21]

Materials Molecular mass Quantity mole Equivalent Intermediate production 318 0.116 g 0.3647 mmol 1 TEA 101.2 0.508 ml 3.647 mmol 10 PCIPCF 190 0.1 ml 0.7194 mmol 2 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone was dissolved in 5 mL of dichloromethane under a nitrogen blanket. TEA and DMAP were added in one portion. A solution of 4-chlorophenylchloroform in 5 mL of dichloromethane was added dropwise at room temperature under a nitrogen blanket. The reaction mixture was stirred at room temperature for 24 h, then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel, overall yield 79%. 68 mg of the desired compound was obtained. MS=472.1 Analysis Calcd (%): C 66.04 H 3.62 N 5.92

Found (%): C 66.21 H 3.60 N 6.04

Example 22: cis-1-[4-(chloro)phenoxycarbonyl]-3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound of Formula P22]

The product was obtained by the procedure of Example 21, except that the trans isomer was dissolved from the silica gel. 68 mg of product was obtained. MS=472.1

Analysis

Calculated (%): C 66.04

Found (%): C 66.19

H 3.62 N 5.92

H 3.57 N 6.04

Example 23: Cis-1-phenoxycarbonyl-3-(2-phthalimido)-4-[2-(phenyl)-ethen-1-yl]azetidinone [Compound of Formula P23]

Materials Molar mass Quantity mole Equivalent Intermediate production 318 0.2g 0.629 mmol 1 PCF 156.6 0.16 ml 1.26 mmol 2 LHMDS 1.0M 0.63ml 0.629 mmol 1

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethen-1-yl]azetidinone was mixed with 6 mL of dry tetrahydrofuran. The mixture was cooled to -75°C. A 1.0 M solution of LHMDS (lithium bis(trimethylsilyl)amide) in tetrahydrofuran was added dropwise via syringe under a nitrogen blanket. The reaction mixture was stirred at -75°C for 30 min. Phenyl chloroformate (PCF) was then added in one portion and the reaction mixture was stirred at room temperature for 1 h. The reaction was then quenched with ammonium chloride solution and extracted with ethyl acetate. The organic phase was separated and purified on silica gel. 0.105 g of a white solid was obtained. Yield = 38.1% MS=438 Analysis Calcd (%): C 71.23 H 4.14 N 6.39

Found (%): C 71.53 H 4.27 N 6.49

Example 24: cis-1-phenoxycarbonyl-3-(2-phthalimido)-4[2-(phenyl-1-yl]azetidinone [Compound of Formula P24]

Materials Molecular mass Quantity mole Equivalence Intermediate production 316 0.1g 0.3164 mmol 1 PCF 156.6 0.08 ml 0.6328 mmol 2 TEA 101.2 0.441 ml 3.164 mmol 10 DMAP 122.2 catalytic

The intermediate 3-(2-phthalimido)-4-[2-(phenyl)ethyn-1-yl]azetidinone was dissolved in dichloromethane at room temperature. TEA and DMAP were added in one portion. A solution of phenyl chloroformate (PCF) in dichloromethane was added dropwise to the reaction mixture at room temperature under a nitrogen blanket. The reaction mixture was stirred at room temperature for 72 h and quenched with 1 N hydrochloric acid. The organic phase was purified on silica gel. 83 mg of a white solid was obtained. Yield = 60.1% MS=436

Example 25: cis-1-phenoxycarbonyl-3-(2-phthalimido)-4-phenylazetidinone [Compound of Formula P25]

Materials Molecular mass Quantity mole Equivalence public sector 296 0.1g 0.3424 mmol 1 TEA 101.2 0.48ml 3.424 mmol 10 PCF 156.6 0.085 ml 0.6848 mmol 2 DMAP A 122.2 3-(2-phthalic m time)- catalytic -4-phenylazetidinone intermediate

dissolved in 5 ml of dichloromethane at room temperature. TEA and DMAP were added in one portion. Phenyl chloroformate (PCF) was then added dropwise and the reaction mixture was stirred at room temperature overnight, then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 48 mg of a white solid was obtained. Yield = 33.9% MS=412 *

Example 26

-phenoxycarbonyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[2-(3-pyridyl)ethen-1-yl]azetidinone [compound of formula P26]

Materials Molar mass Quantity mole Equivalence Intermediate production 333 0.1g 0.6006 mmol 1 TEA 101.2 0.167 mmol 1,2012 mmol 2 PCF 156.6 0.150 m1 1,2012 mmol 2 DMAP 122.2 catalytic

The intermediate 3-[4S-phenyloxazolidin-2-on-3-yl]-4-[2-(3-pyridyl)ethen1-yl]azetidinone was dissolved in dichloromethane. The reaction mixture was cooled to -10°C under a nitrogen blanket. At this temperature, DMAP was added followed by TEA. The reaction mixture was stirred for 5 minutes and phenyl chloroformate was added dropwise via syringe. After 1 hour of addition, the reaction was complete by TLC. The reaction mixture was stirred for a further 1 hour at +10°C and quenched with saturated ammonium chloride solution. The organic phase was dried over magnesium sulfate and evaporated under reduced pressure. The residue was then suspended in ether at room temperature. The mixture was filtered and 87 mg of an almost white solid was collected. Analysis Calculated (%): C 68.56 H 4.65 N 9.23

Found (%): C 68.45 H 4.74 N 9.04

Example 27

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone hydrochloride [Compound of formula P27]

87 g of the free base of the product of Example 36 was dissolved in ethyl acetate. 5 equivalents of hydrochloric acid in 1N ether were added dropwise at room temperature. A precipitate formed immediately. The reaction mixture was stirred at room temperature for 30 minutes. The solvents were removed under reduced pressure to give 105 mg of a white solid. MS = 455.

Analysis

Calcd (%): C 63.48 H 4.51 N 8.54 Cl 7.21

Found (%): C 63.27 H 4.46 N 8.44 Cl 6.91

Example 28

-phenoxycarbonyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3pyridyl)ethen-1-yl]azetidinone hydrochloride [Compound of formula P28]

Materials Molar mass Quantity mole Equivalent Intermediate production 333 0.1g 0.3003 mmol 1 PCF 156.6 0.075 ml 0.6006 mmol 2 TEA 101.2 0.4186 ml 3.003 mmol 10 Materials Molecular mass Quantity mole Equivalent DMAP 122.2 catalytic The 3-[4S-phenyl -oxazolidine-2 1-on-3-yl]-4R-[2-(3-pyridyl)-

The intermediate [[(ethen-.1-yl]azetidinone]] was dissolved in dichloromethane at room temperature under a nitrogen blanket. TEA and DMAP were added in one portion. A solution of phenyl chloroformate (PCF) in dichloromethane was added dropwise to the reaction mixture under a nitrogen blanket. The reaction mixture was stirred at room temperature overnight and then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 43 mg of product was obtained as a 1:1 cis/trans mixture.

Example 29

1-phenoxycarbonyl-3S-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3pyridyl)ethen-1-yl]azetidinone hydrochloride [Compound P29]

The product was obtained by the procedure of Example 28 except that the cis isomer was separated. 30 mg of product was obtained. MS=491

Example 30

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[(2-quinolinyl)ethen-1-yl]azetidinone hydrochloride [Compound with formula P30]

Materials Intermediate prod. Molar mass Quantity mole Equivalent 335 0.157 g 0.4686 mmol 1 Materials PCF Molarityeq Quantity mol Equivalent 156.6 0.118 ml 0.9377 mmol 2 TEA 101.2 0.635 mL 4.686 mmol 10 DMAP 122.2 catalytic The 3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[(2-quinolinyl)-

The ethen-1-ylazetidinone intermediate was dissolved in 5 mL of dichloromethane at room temperature. TEA and DMAP were added in one portion, followed by phenylchloroformate (PCF) dropwise at room temperature. The reaction mixture was stirred at room temperature for 16 h, then quenched with 1 N hydrochloric acid. The organic phase was separated and 5 equivalents of 1 N ethereal hydrochloric acid were added. 43 mg of a white solid was obtained. Yield = 16.3% MS = 528

Example 31

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yl]azetidinone [Compound with formula P31]

Materials Molecular mass Quantity mole Equivalence Intermediate production 334 0.166g 0.497 mmol 1 TEA 101.2 0.416 ml 2.98 mmol 6 PCF 156.6 0.125 ml 0.994 mmol 2 DMAP 122.2 catalytic

The intermediate 3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethenyl]-azetidinone was dissolved in 5 mL of dichloromethane at room temperature. TEA and DMAP were added in one portion, followed by phenyl chloroformate (PCF) dropwise. The reaction mixture was stirred at room temperature for 24 h, then quenched with 1 N hydrochloric acid. The organics were separated and purified on silica gel. 60 mg of the trans isomer was obtained.

Example 32

-phenoxycarbonyl-3S-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yl]azetidinone [Compound of formula P32]

Following the procedure of Example 30, 55 mg of the cis isomer was dissolved from the silica gel.

Example 33

-phenoxycarbonyl-3-(2-phthalimido)-4-(3-buten-1-ylazetidinone [compound with formula P33]

Materials Molecular mass Quantity mole Equivalent Intermediate production 270 0.2 g 0.741 mmol 1 PCF 156.6 0.186 ml 1.48 mmol 2 TEA 101.2 1.03 ml 7.407 mmol 10 DMAP 122.2 catalytic The 3-(2-phthalim time) -4-(3-buten-1-ylazetidinone intermediate

product is mixed with 20 ml of dichloromethane at room temperature. TEA and DMAP are added in one portion. A solution of PCF in 10 ml of dichloromethane is added dropwise to the reaction mixture at room temperature under a nitrogen blanket. The reaction mixture is stirred for 24 hours, then the reaction is quenched with 1 N hydrochloric acid. The reaction mixture is purified on silica gel to give the pure product, 108 mg of a white solid (1:1 cis/trans mixture). Yield = 37% MS=390

34; Example

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone acetate [Compound of formula P34]

1 g of the azetidinone obtained in Example 26 was dissolved in 10 ml of ethyl acetate, followed by the addition of 1 ml of acetic acid. The reaction mixture was stirred at room temperature for 5 hours, filtered, and the white solid (0.75 g) was ignited. Yield = 66.2% MS=455

Example 35

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-pyridyl)ethen-1-ylazetidinone [Compound of formula P35]

Materials Molar mass Quantity mole Equivalent Intermediate production 333 0.1g 0.6006 mmol 1 TEA . 101.2 0.167 ml 1,2012 mmol 2 PCF 156.6 0.150 ml 1,2012 mmol 2 DMAP 122.2 catalytic

The intermediate 3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-pyridyl)ethen1-yl]azetidinone was dissolved in 5 mL of dichloromethane at room temperature. TEA and DMAP were added in one portion, followed by phenyl chloroformate dropwise. The reaction mixture was stirred at room temperature for 16 h, then quenched with 1 N hydrochloric acid. The organic phase was separated and 5 equivalents of 1 N ethereal hydrochloric acid were added. The mixture was filtered to give 63 mg of a white solid.

Yield = 21.4% MS=455.

Example 36

-phenoxycarbonyl-3R-[4R-phenyloxazolidin-2-on-3-yl]-4S-[2(phenyl)ethen-1-yl]azetidinone [compound of formula P36]

Materials Molar mass Quantity mole Equivalent Intermediate production 334 32.9 mg 0.0985 mmol 1 PCF 157 0.025 ml 0.197 mmol 2 TEA 101.2 0.137 ml 0.985 mmol 10 DMAP 122.2 catalytic

The intermediate 3R-[4R-phenyloxazolidin-2-on-3-yl]-4S-[2-(phenyl)ethen-1 yl]azetidinone was dissolved in 2 mL of dichloroethane at room temperature. The reaction mixture was protected with a tube containing a desiccant. DMAP and TEA were added in one portion, followed by PCF via syringe, and the reaction mixture was stirred at room temperature. TLC indicated that the reaction was complete within a few minutes. The reaction was quenched with ammonium chloride. The product was purified on silica gel to give 28 mg of pure trans product as a white solid. Yield = 63% MS = 454

Example 37 1-Phenoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3pyridyl-N-oxido)ethen-1-yl]azetidinone [Compound of formula P37] Substances Molar mass Quantity mol Equivalent Int. yield 455 0.138 g 0.3032 mmol 1

MCPBA 172.6 0.115 g 0.6670 mmol 2.2

The intermediate 3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl-Noxydo)ethen-1-yl]azetidinone was dissolved in 10 ml of dichloromethane at room temperature. MCPBA (3-chloroperoxybenzoic acid) was added in one portion and the reaction mixture was stirred at room temperature for 16 hours. The reaction was quenched with 10% sodium thiosulfate solution. The organic phase was separated and the resulting mixture was dried over magnesium sulfate and filtered. The organic phase was evaporated to give a white foam. ¹ H-NMR spectrum and TLC showed the presence of the N-oxide. The product was obtained as 105 mg of a white powder. Yield = 73.5% MS=471

Example 38

-phenoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(4-pyridyl)ethen-1-yl]azetidinone hydrochloride [Compound of formula P38]

Materials Molar mass Quantity mole Equivalent Intermediate production 333 606 mg 1.82 mmol 1 PCF 156.6 0.457 ml 3.64 mmol 2 TEA 101.2 0.507 ml 3.64 mmol 2 DMAP 122.2 catalytic The 3R-[4S-phenyl- -oxazolidin2-on-3-yl]-4R-[2-(4-pyridyl)-

The intermediate [[(ethen-1-yl]azetidinone]] was mixed with 10 mL of dichloromethane at room temperature. TEA and DMAP were then added in one portion. Phenyl chloroformate (PCF) in approximately 10 mL of dichloromethane was slowly added dropwise to the reaction mixture protected by a tube containing a drying agent at room temperature. The reaction was complete by TLC. The reaction mixture was applied directly to a silica gel column and purified with hexane (1:1 hexane/ethyl acetate). The solid was dissolved in ethyl acetate and 5 equivalents of 1N hydrochloric acid in ether were added dropwise. The reaction mixture was stirred at room temperature for 1 h and then filtered. 202 mg of an off-white solid was collected. MS = 455

Analysis

Calcd (%): C 68.56 H 4.65 N 9.23

Found (%): C 68.80 H 4.70 N 9.48

Example 39

1-[4-methoxycarbonylphenoxycarbonyl]-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yl]azetidinoneJP39 compound]

Materials Molecular mass Quantity mole Equivalent Intermediate production 334 248 mg 0.7425 mmol 1 MCPCF 214.6 175 mg 0.817 mmol 1.1 TEA 101.2 0.207 ml 1.485 mmol 2 DMAP 122.2 catalytic

The intermediate 3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1yl]azetidinone was dissolved in 5 mL of dichloromethane at room temperature. DMAP was added to the reaction mixture followed by TEA. A solution of 4-methoxycarbonylphenylchloroform (MCPCF) in 10 mL of dichloromethane was added dropwise at room temperature. The reaction mixture, protected by a tube containing a drying agent, was stirred at room temperature overnight and then quenched with 1 N hydrochloric acid. The organic phase was separated and purified on silica gel. 274 mg of a white material was obtained. Yield = 72% MS = 512

Example 40

-[(4-chlorobutyl)oxycarbonyl]-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yl]azetidinone [Compound with formula P40]

Materials Molar mass Quantity mole Equivalent Intermediate production 334 43.2 mg 0.1293 mmol 1 TEA 101.2 0.18 ml 1.293 mmol 10 4-BCF 171 0.035 ml 0.2586 mmol 2 DMAP 122.2 catalytic

The intermediate 3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1 yl]azetidinone was dissolved in dichloromethane at room temperature. DMAP and TEA were added via syringe, followed by chlorobutyl chloroformate (4-BCF) dropwise. The reaction mixture was stirred with dry ice at room temperature, then the reaction mixture was quenched with 1 N hydrochloric acid, and the organic phase was separated and purified on silica gel. 32.77 mg of product was obtained. Yield = 64% MS=468

41, Example

-phenoxycarbonyl-3-[4R-phenyloxazolidin-2-on-3-yl]-4S-[2(phenyl)ethen-1-yl]azetidinone [Compound with formula P41]

Materials Molecular mass Quantity mole Equivalent Intermediate production 334 0.1g 0.244 mmol 1 PCF 157 0.075 ml 0.599 mmol 2 TEA 101.2 0.42 ml 2.994 mmol 10 DMAP 112.2 catalytic The 3R-[4R-phenyl- oxazolidin-2-on-3-yl]-4S-[2(phenyl)ethene-1-

The intermediate [[(

Example 42

-phenoxycarbonyl-3-[4R-phenyloxazolidin-2-on-3-yl]-4S-[2(phenyl)ethen-1-yl]azetidinone [compound with formula P42]

In the process of Example 41, 23 mg of cis-configuration, optically active product is obtained from silica gel. MS=454

Example 43: cis-1-phenoxycarbonyl-3(2-phthalimido)-4-buten-1-yl-azetidinone

[Compound with formula P43]

In the procedure of Example 33, 15 mg of pure cis isomer is isolated from silica gel. MS=390

Example 44: cis-1-phenoxycarbonyl-3-[oxazolidin-2-on-3-yl]-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound P44]

Materials Molecular mass Quantity mole Equivalence Intermediate production 258 0.44g 0.0017 mole 1 LHMDS/THF 1 N 1.7ml 0.00187 moles 1.1 PCF 156.6 0.32 ml 0.00255 mole 1.5

The 3-(oxazolidin-2-on-3-yl)-4-[2-(phenyl)ethen-1-yl]acetic intermediate was dissolved in 17 mL of tetrahydrofuran. The reaction mixture was maintained at a temperature of about -73°C to about -78°C using a dry ice/acetone cooling mixture. A 1.7 M solution of LHDMS in tetrahydrofuran was added dropwise to the reaction mixture via syringe. 0.32 mL of PCF was added in one portion and the reaction mixture was stirred until all reactants were consumed. The reaction was then quenched with 20 mL of 0.5 N hydrochloric acid and extracted with 20 mL of diethyl ether and 30 mL of dichloromethane. A solid formed and was dissolved in 50 mL of dichloromethane. The combined organic layers were first

Wash with 10% sodium bicarbonate, then with 50 ml of 12.5% sodium chloride and dry over magnesium sulfate. The mixture is then filtered and evaporated. The residue is dissolved in 20 ml of dichloromethane and added to 20 ml of ice-cold diethyl ether. The white solid is filtered under reduced pressure, washed with 10 ml of diethyl ether and dried under reduced pressure. MS=378.38

Example 45: trans-1-phenoxycarbonyl-3R-[5-(isopropyl)oxazolidin-2-on-3-yl]4R-[2-(phenyl)ethen-1-yl]azetidinone [Compound P45]

Materials Molar mass Quantity mole Equivalence Intermediate production 300.36 0.11g 0.00036 moles 1 LHMDS/THF 1N 0.40 ml 0.0004 moles 1.1 PCF 156.6 0.07 ml 0.005 mole 1.5

The intermediate 3-[(5-isopropyl)oxazolidin-2-on-3-yl]-4-(2(phenyl)ethen-1-yl)azetidinone was dissolved in 10 mL of tetrahydrofuran. The temperature of the reaction mixture was maintained between about -73°C and about -78°C using a dry ice/acetone cooling mixture. 0.40 mL of a 1 N solution of LHMDS in tetrahydrofuran was added dropwise via syringe. 0.07 mL of PCF was added in one portion and the reaction mixture was stirred until all the starting material was consumed. The reaction mixture was removed from the ice bath and the reaction was quenched with 20 mL of 0.5 N hydrochloric acid followed by two 20 mL portions of dichloromethane. The dichloromethane portion was first extracted with 20 mL of 10% sodium bicarbonate solution and then with 20 mL of The mixture is washed with 12.5% sodium chloride solution and dried over magnesium sulfate. The mixture is then filtered and evaporated. Chromatography of the residue in dichloromethane on silica gel gives the product.

Example 46: cis-1-phenoxycarbonyl-3-[(5-isopropyl)oxazolidin-2-on-3-yl]-4-[2(phenyl)ethen-1-yl]azetidinone_[Compound of formula P46]

Materials Molecular mass Quantity mole Equivalent Intermediate production 300.36 0.1g 0.00033 mole 1 LFMDS/THF 1N 0.37ml 0.00036 mole 1.1 PCF 156.6 0.06 ml 0.005 mole 1.5

The intermediate 3-[(5-isopropyl)oxazolidin-2-on-3-yl]-4-[2(phenyl)ethen-1yl]azetidinone was dissolved in 10 mL of tetrahydrofuran. The temperature of the reaction mixture was maintained between about -73°C and about -78°C using a dry ice/acetone cooling mixture. 0.40 mL of a 1 N solution of LHMDS in tetrahydrofuran was added dropwise via syringe. 0.07 mL of PCF was added in one portion and the reaction mixture was stirred until all the starting material was consumed. The reaction mixture was removed from the ice bath and the reaction was quenched with 20 mL of 0.5 N hydrochloric acid followed by extraction with 20 mL of dichloromethane. The dichloromethane portion is washed first with 20 ml of 10% sodium bicarbonate solution, then with 20 ml of 12.5% sodium chloride solution and dried over magnesium sulfate. The mixture is then filtered and evaporated. The residue is dissolved in 20 ml of dichloromethane and added dropwise, in an ice bath, to a mixture of 10 ml of diethyl ether and 10 ml of hexane. A white solid is separated, which is filtered under reduced pressure. MS=420.3

Reaction Scheme II below illustrates the preparation of the compounds of Examples 47-50.

The bases and solvents used to carry out the reactions described in Preparation Scheme II are the same as those used in Preparation Scheme I.

The following is a more detailed description of the reaction steps depicted in Preparation Scheme II. It will be apparent to those skilled in the art that these reaction steps are chemically only minor modifications of the steps of Preparation Scheme I. The following descriptions and examples are intended to further illustrate the preparation of the compounds of the present invention and are not intended to limit the scope of the invention in any way. Other methods of preparing the compounds of the present invention, although not disclosed herein, are intended to be within the scope of the present invention.

The starting aldehyde of formula (i) is prepared as described in detail in Tet. Lett., 32 (6), 803-6 (1991). Acid chloride dissolved in solvent is added dropwise to a mixture of the appropriately substituted imine, solvent, and amine to give the starting aldehyde of formula (i). The reaction mixture is stirred at room temperature under an inert gas blanket until the reaction is complete. Then, 5% hydrochloric acid solution is added and the reaction mixture is stirred for about 1.5 hours. A solvent is added to the organic layer, the layer is washed with 5% hydrochloric acid, then with water and saturated sodium bicarbonate solution. The resulting product is dried and the solvent is removed under reduced pressure. The preferred solvent used in the reaction is toluene, and the preferred amine is triethylamine (TEA). The appropriately substituted olefin can then be prepared from the aldehyde of formula (i) by either the Wittig reaction or the HornerEmmons reaction, as described in Orq. React., 14, 270 (1965), and Acc. Chern. Res., 16, 411 (1983). [Step 1] Alkene derivatives are prepared from carbonyl compounds by the Wittig reaction, which involves reacting the starting compound with an alkyltriphenylphosphanium salt in the presence of a base, or by the WadsworthEmmons reaction (a Horner-Emmons modification of the Wittig reaction), in which phosphonate esters are reacted with carbonyl derivatives in the presence of a base to give the alkenes. Oxidation with CAN [Step 2] and acylation [Step 3] Reaction Step] is the same as Reaction Steps 5A and 6A, or 4B and 5B of Preparation Scheme I.

Example 47

1-phenoxycarbonyl-3-benzyl-4-[2-(phenyl)ethen-1-yl]azetidinone

[Compound with formula P47]

Materials Molecular mass Quantity mole Equivalent Intermediate production 263 223 mg 0.8479 mmol 1 LHMDS/THF 1.0 M 0.8479 ml 0.8479 mmol 1 PCF 156.6 0.212 ml 1.6953 mmol

The intermediate 3-benzyl-4-[2-(phenyl)ethen-1-yl)azetidinone was dissolved in 8 mL of dry tetrahydrofuran. The reaction mixture was cooled to -78°C and a 1.0 M solution of LHDMS in tetrahydrofuran was added dropwise via syringe under a nitrogen blanket. The reaction mixture was stirred at -78°C for 30 min. The PCF was added at -78°C. The reaction mixture was stirred for 30 min, the cooling bath was removed and the reaction was quenched with saturated ammonium chloride solution. The reaction mixture was stirred at room temperature and purified on silica gel. 237 mg of a white solid was obtained. Yield = 73%

%.···:·..»··..·

Analysis

Calculated (%): C 78.31 H 5.52

Found (%): C 78.05 H 5.68

N 3.65

N 3.46

Example 48: cis-1-phenoxycarbonyl-3-benzyl-4-[2-(3-pyridyl)ethen-1-yl]azetidinone hydrochloride [Compound P48]

Materials Molar mass Quantity mole Equivalent Intermediate production 264 0.1g 0.379 mmol 1 TEA 101.2 0.53 ml 3.787 mmol 10 PCF 156.6 0.095 ml 0.7574 mmol 2 DMAP 122.2 catalytic

The intermediate cis-1-benzyl-[2-(pyridin-3-yl)ethen-1-yl]azetidinone was dissolved in 5 mL of dichloromethane at room temperature. TEA and DMAP were added in one portion. Then a solution of phenyl chloroformate was added dropwise. The solution was stirred at room temperature overnight and the reaction was quenched with 1 N hydrochloric acid. The organic phase was separated and the resulting product was purified on silica gel. 78 mg of a white solid was obtained. Yield = 48.9% MS=421

Example 49: cis-1-phenoxycarbonyl-3-benzyl-4-[2-(cyano)ethen-1-yl]azetidinone [Compound of formula P49]

Materials Molar mass Quantity mole Equivalent Intermediate production 212 0.1 g 0.4716 mmol 1 PCF 156.6 0.118 ml 0.9432 mmol 2 LHMDS 1.0 M 0.4716 ml 0.4716 mmol 1 The 3-benzyl-4-[2- (cyano)ethen-1-yl]azetidinone intermediate

product is mixed with 4 ml of tetrahydrofuran under a nitrogen blanket at a temperature of about -70°C. A 1.0 M solution of LHDMS in tetrahydrofuran is added dropwise to the reaction mixture via syringe. The reaction mixture is stirred at a temperature of about -70°C for 30 minutes, then PCF is added via syringe. The reaction mixture is stirred at a temperature of about -70°C for an additional 30 minutes, then the reaction is quenched with 1N hydrochloric acid. The organic phase is separated and purified on silica gel. 105 mg of product is obtained. Yield = 67% MS=332 Analysis

Calcd (%): C 72.28 H 4.85 N 8.43

Found (%). C 72.02 H 5.05 N 8.38

Example 50: trans-1-phenoxycarbonyl-3-benzyl-4-[2-(phenyl)ethen-1-yl]azetidinone [Compound P50]

The trans isomer is isolated in Example 47.

The present invention provides the following new and advantageous method

I also provides for the preparation of certain azetidinone derivatives characterized by the general formula I, and also provides intermediates for the preparation of azetidinone derivatives. More specifically, the invention relates to the individual reaction steps and the overall process for preparing the compound of formula Ia, or its pharmaceutically acceptable salts and solvates, wherein the group of formula R ¹⁶ is phenyl, 2-nitrophenyl, 4-nitrophenyl, naphthyl, 3-nitro-1-naphthyl, 4-nitro-2-naphthyl, 2-quinolinyl, 4-quinolinyl, 7-quinolinyl, 1-isoquinolinyl, 3-isoquinolinyl, 8-isoquinolinyl, 2-benzothiazolyl, 2-benzoxazolyl, 2-thienyl, 3-furanyl, 3-pyridinyl, 3-pyridinyl-1-N-oxide, 5-pyrimidinyl, or 3,5-dimethylphenyl; and

R ¹⁷ is 1-phenoxycarbonyl, 4-chlorophenoxycarbonyl, 4-fluorophenoxycarbonyl, benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-chlorobutyloxycarbonyl, 4-methoxycarbonylphenoxycarbonyl, 4-methoxyphenoxycarbonyl or 1-propylenoxycarbonyl;

according to which process an aldehyde of general formula II, in which the group of general formula R ¹⁸ is a trialkylsilanyl group, and an amine of general formula III, protected on the nitrogen atom, in which the group of general formula R ¹⁹ is a nitrogen protecting group, are reacted to produce an imine of general formula IV, and then the reaction of the imine of general formula IV, protected on the nitrogen atom, and 4S-phenyl-oxazolidin-2-on-3-yl-acetyl chloride of formula V produces an azetidinone derivative of general formula VI, protected on the nitrogen atom; desilylation and epimerization of the nitrogen-protected azetidinone derivative of formula VI to give the desilylated azetidinone intermediate of formula VII substituted with an acetylene group at the 4-position, reduction of this intermediate and removal of the nitrogen-protecting group to give the 3-(4S-phenyloxazolidin-2-on-3-yl)-4-ethenyl azetidinone of formula IX; reaction of the compound of formula IX with an electrophilic reagent of formula XR ¹⁶ (where X is a leaving group) in the presence of a catalyst to give the alkenyl-substituted azetidinone derivative of formula XI, and finally acylation of the compound of formula XI to give the substituted oxycarbonyl azetidinone derivative of formula Ia.

Preferred intermediates of the present invention are compounds of formula VIII, wherein ^R19 is a nitrogen protecting group, and compounds of formula IX.

Scheme III further illustrates the preferred method for preparing certain azetidinone derivatives of the present invention. In Scheme III, R ¹⁶ , R ¹⁷ , R ¹⁸ , and R ¹⁹ are as previously defined.

The nitrogen-protected imine of the formula (IV) is prepared by methods known per se, for example, by reacting an aldehyde of the formula (II) and a primary amine of the formula (III) protected at the nitrogen atom, such as ρ,-anisidine, in the presence of a solvent, preferably methylene chloride or dichloromethane [Reaction step 1]. The aldehyde of the formula (II) can be prepared by methods known per se, for example, as described in Hauptman, H. and Mader, M., Synthesis, 307 (1978). Suitable R ¹⁸ groups are, for example, trimethylsilyl or other alkylsilyl groups. In addition, various known nitrogen-protecting groups of the formula R ¹⁹ can be used. A drying agent is advantageously added to the reaction mixture, which is typically magnesium sulfate, sodium sulfate, silica gel, or the like. The reaction mixture is stirred at room temperature overnight until the reaction is complete, as determined by a readily available method. The reaction mixture is then filtered and the solvent is removed under reduced pressure to give the desired nitrogen-protected imine of formula (IV).

The reaction of 4S-phenyloxazolidin-2-on-3-ylacetyl chloride of formula (V) with the nitrogen-protected imine of formula (IV) produces the nitrogen-protected azetidinone derivative of formula (VI), mainly in the form of the cis isomer [Reaction Step 2]. The 2+2 cycloaddition method is generally well known in the art. See: G.I. Georg, Ed.; Organic Chemistry of β-Lactams, Chapter 6, Verlag Chemie, New York, (1992); United States Patent Applications Nos. 4,751,299 and 4,665,171, which are incorporated herein by reference. One equivalent of 4S-phenyloxazolidin-2-on-3-ylacetyl chloride [compound (V)] and a solvent, such as dichloromethane, are charged to a flask and the mixture is cooled to about -78°C. A suitable base, such as a tris(C1-4 alkyl)amine, typically triethylamine, is then added to the mixture while maintaining the temperature of the mixture in the vicinity of -70°C. The reaction mixture is stirred thoroughly. The imine dissolved in the solvent is then slowly added at -70°C. Although the time of addition is not critical, best results are obtained if the addition is made dropwise over a period of about 1 to about 6 hours and the reaction mixture is stirred for about 2 to 4 hours. The reaction is carried out at a temperature of about -78°C to about 25°C, preferably about -78°C to about 0°C, more preferably about -78°C to -75°C, in a solvent in the presence of tris(C1-C4 alkyl)amine.

4S-Phenyl-oxazolidin-2-on-3-yl-acetyl chloride [compound (V)] can be prepared by methods known per se, such as the process described in U.S. Patent Application No. 4,665,171, which is incorporated herein by reference. For example, to prepare the acid chloride, 1.8 equivalents of oxalyl chloride are reacted with one equivalent of the appropriately substituted acid in the presence of a solvent, preferably toluene or benzene. The reaction mixture is preferably heated to about 60°C for about 3-4 hours under an inert gas blanket, such as nitrogen, helium or argon. The reaction mixture is cooled to room temperature and the solvent is removed under reduced pressure.

Then, by desilylation of the nitrogen-protected azetidinone derivative [compound of general formula (VI)] and epimerization at the 3-position, a 4:1 mixture of trans:cis isomers of the nitrogen-protected, desilylated, 4-acetylenically substituted azetidinone intermediate [Reaction step 3]. The desilylation and epimerization of the nitrogen-protected azetidinone derivative [compound of general formula (VI)] can be carried out by various methods known per se. However, in a favorable case, the desilylation and epimerization of the compound of general formula (VI) is carried out with tetrabutylammonium fluoride (TBAF) at a temperature below -50°C. A favorable solvent for the reaction is tetrahydrofuran.

Partial reduction of the desilylated, 4-acetylenically substituted trans-azetidinone intermediate [compound of formula (VII)] and removal of the nitrogen protecting group affords the trans-3-(4S-phenyl-oxazolidin-2-on-3-yl)-4-ethenyl-azetidinone β-lactam intermediate [compound of formula (IX), Reaction Steps 4 and 5]. The partial reduction is conveniently carried out by hydrogenation of compound (vii) with Lindlar catalyst and quinoline. The nitrogen protecting group is conveniently removed with cerium ammonium nitrate in acetonitrile, for example, according to the method described in J. Orq. Chem., 47, 276568 (1982). It will be apparent to those skilled in the art that the partial reduction and removal of the protecting group can be carried out in any order.

The resulting trans-3-(4S-phenyloxazolidin-2-on-3-yl)-4ethenylazetidinone [compound (IX)] is first functionalized by reacting with an electrophilic reagent of formula XR ¹⁶ (where X is a leaving group) in the presence of a solvent, and then acylating the resulting compound to obtain the substituted oxycarbonylazetidinone derivative of formula I [Reaction steps 6 and 7]. In formula XR ¹⁶ , X is a leaving group such as a halide, sulfonate, or the like, and R ¹⁶ is as defined above. The functionalization is most advantageously accomplished with the aid of a catalyst such as palladium acetate, preferably in the presence of potassium acetate dissolved in dimethylformamide and tetrabutylammonium chloride. See A. Satake, et al., Synlett, 839 (1994). Preferred ^R groups are 2-nitro-4phenyl, 4-nitro-2-phenyl, 3-nitro-1-naphthyl, 4-nitro-2-naphthyl, 2-quinolinyl, 4-quinolinyl, 7-quinolinyl, 1-isoquinolinyl, 3-isoquinolinyl, 8-isoquinolinyl, 2-benzothiazolyl, 2-benzoxazolyl. More preferred R groups are 2-thienyl, 3-furanyl, 5-pyrimidinyl, and 3,5-dimethylphenyl. It is obvious to those skilled in the art that the functionalization of the trans-3-(4S-phenyl-oxazolidin-2-on3-yl)-4-ethenyl-azetidinone of formula (IX) or the acylation of the compound can be carried out in any order, although the former reaction with the reagent of general formula XR ^1S , the product of which is the alkenyl-substituted azetidinone derivative of general formula (XI), advantageously precedes the acylation.

The acylation is advantageously carried out with a suitable chloroformate to give the substituted oxycarbonylazetidinone derivative of formula Ia. Suitable chloroformates include phenyl chloroformate, 4-chlorophenyl chloroformate, 4-fluorophenyl chloroformate, benzyl chloroformate, 4-nitrobenzyl chloroformate, 4-chlorobutyl chloroformate, 4-methoxycarbonylphenyl chloroformate, 4-methoxyphenyl chloroformate, and allyl chloroformate.

Scheme III illustrates the preferred sequence of reaction steps of the present invention. However, compounds of formula Ia may be prepared by altering the order of certain reaction steps. For example, compounds of formula Ia may be prepared by reducing an acetylenically substituted intermediate of formula VII to compound of formula VIII, followed by

1- by removing the protecting group from the compound of formula VIII to form a compound of formula IX, followed by reaction with the electrophilic reagent of formula XR ¹⁶ and acylation (in any order); or

2- reaction of compound VIII with electrophilic reagent XR ¹⁶ followed by deprotection and acylation. It is possible to first deprotect the azetidinone intermediate VII containing an acetylene group at the 4-position, then reduce to give compound IX followed by reaction with electrophilic reagent XR ¹⁶ followed by acylation (in any order).

The following Preparations and Examples further illustrate the preparation of the compounds of the present invention and are not intended to limit the scope of the invention in any way.

Preparation of compound of formula IVa [(J) Reaction]

4.89 g of the aldehyde of formula (Ha) are dissolved in 10 ml of methylene chloride. 9.43 g of anhydrous magnesium sulfate are added, followed by a solution of 4.77 g of p-anisidine in 20 ml of methylene chloride. The reaction mixture is stirred at room temperature for 90 minutes, then filtered, and a brown liquid is obtained by evaporating the solvent. The material is dissolved in hexane. The hexane solution is poured off the insoluble residue, then washed with 1% hydrochloric acid solution. After drying over magnesium sulfate, 7.36 g of the imine is obtained by evaporating the solvent, as an orange liquid.

TLC: R _f 0.74 (3/1 hexane/ethyl acetate);

¹ H-NMR spectrum spectrum (CDCI ₃ , δ, 500 MHz): 7.73 (s, 1H), 7.20 (d, 1H, J = 8.9 Hz), 6.91 (d, 1H, J = 8.9 Hz), 3.83 (s, 3H), 0.29 (s, 9H).

Preparation of compound of formula V [(K) Reaction] 4S-Phenyl-oxazolidin-2-on-3-yl-acetyl chloride

100 g of the acid of formula Va are suspended in toluene at room temperature. 103.6 g of oxalyl chloride are added in one portion. The solution is heated to 60°C for 4 hours while purging with nitrogen. The reaction mixture is then cooled to room temperature and the toluene is removed under reduced pressure to give an oil. The oil solidifies on standing below 0°C. 113 g of the acid chloride are thus obtained.

Preparation of the compound of formula Via [(L) Reaction]

-methoxyphenyl-3-[4S-phenyloxazolidin-2-on-3-yl]-(2-trimethylsilyl)ethyn-1-ylazetidinone

The 2+2 cycloaddition is carried out by dissolving 3.4 g of the acid chloride of formula V in 40 ml of dichloromethane and cooling the solution in a dry ice/acetone bath. 3.0 ml of triethylamine is added dropwise, which turns the solution violet. After 15 min, the imine of formula IVa (3.6 g in 24 ml of toluene) is added dropwise. After 15 min, the dry ice/acetone bath is replaced with an ice bath. After 3 h, the solution is warmed to room temperature. The reaction mixture is diluted with ethyl acetate, then washed with 10% hydrochloric acid solution (3x), saturated sodium chloride solution (1x), saturated sodium bicarbonate solution (2x), and finally saturated sodium chloride solution (1x). The organic solution was dried over sodium sulfate and concentrated on a rotary evaporator. The 5.5 g amber solid was recrystallized from 60% ethyl acetate/40% hexane to give 3.72 g of 1-methoxyphenyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(2-trimethylsilylethyl)-1-yl]azetidinone β-lactam.

TLC: R _f 0.25 (2/1 ethyl acetate/hexane);

¹ H-NMR spectrum (CDCl ₃ , 6, 500 MHz): 7.51-7.40 (m, 7H), 6.90-6.87 (m, 2H), 4.99 (t, 1H, J = 8.8 Hz), 4.72-4.68 (m, 2H), 4.42 (d, 1H, J = 5.0 Hz), 4.21 (t, 1H, J = 8.9 Hz), 3.81 (s, 3H), 0.26 (s, 9H).

Preparation of compound of formula VIIa [(M) Reaction]

-methoxy-phenyl-3-[4S-phenyl-oxazolidin-2-on-3-yl]-4-[(2-trimethylsilylethyl-1-yl]azetidinone

6.0 g of 1-methoxyphenyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(2trimethylsilyl)-1-yl]-azetidinone were dissolved in 150 ml of tetrahydrofuran and the solution was cooled in a dry ice/acetone bath. 4.34 g of tetrabutylammonium fluoride were added in three portions. The reaction mixture was allowed to warm slowly to room temperature overnight in the presence of the cold bath. The temperature of the reaction mixture was reduced to 2/3 by evaporation in a rotary flask distillation apparatus. The solution was diluted with ethyl acetate and washed with 10% hydrochloric acid solution (2x), saturated sodium chloride solution (1x), saturated sodium bicarbonate solution (2x), and saturated sodium chloride solution (1x). The organic solution was dried over sodium sulfate and concentrated by rotary evaporation to give 4.43 g of the desired 1-methoxyphenyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(2-trimethylsilyl)ethyn-1-yl]azetidinone β-lactam as a 4.3:1 trans:cis isomer mixture. This material was used in the next step without further purification.

TLC: R _f 0.46 (trans): 0.37 (cis) (1/1 ethyl acetate/hexane);

the spectrum data were obtained from a 4.3:1 trans:cis mixture, the integral values of the overlapping signals were corrected in some cases;

¹ H-NMR spectrum (CDCl ₃ , δ, 500 MHz): trans isomer 7.487.37 (m, 7 H), 6.87 (d, 2 H, J = 8.9 Hz), 5.03-4.99 (m, 1 H), 4.91 (t, 1 H, J = 2.1 Hz), 4.75 (t, 1 H, J = 8.8 Hz), 4.42 (d, 1 H, J = 2.5 Hz), 4.30-4.24 (m, 1 H), 3.78 (s, 3 H), 2.38 (d, 1 H, J = 1.6 Hz); distinct signals of the cis isomer: 4.53 (d, 1 H, J = 4.8 Hz), 2.67 (d, 1 H, J = 1.8 Hz).

Preparation of compound of formula Villa [(N) Reaction]

1-Methoxyphenyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(2-trimethylsilyl)ethen-1-yl]azetidinone

2.0 g of 1-methoxyphenyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(2-trimethylsilyl)ethyn-1-yl]azetidinone were dissolved in a mixture of 60 ml of methanol and 140 ml of dichloromethane. 0.55 ml of quinoline and 0.15 g of Lindlar catalyst were added. The mixture was stirred under hydrogen atmosphere for 5 hours, then filtered through a filter bed containing Celite and the solvent was removed using a rotary distillation apparatus. The residue was dissolved in ethyl acetate and washed with 10% hydrochloric acid solution (3x) and then with saturated sodium chloride solution (1x). The organic solution was dried over sodium sulfate and concentrated in a rotary evaporator to give 2.00 g of the desired 1-methoxyphenyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(2-trimethylsilyl)ethen-1-yl]azetidinone β-lactam as a 4.3:1 trans:cis isomer mixture. This material was used in the next step without further purification.

TLC: R _f 0.41 (trans 0.45 (cis) (1/1 ethyl acetate/hexane);

the spectrum data were obtained from a 4.3:1 trans:cis mixture, the integral values of the overlapping signals were corrected in some cases;

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): trans isomer: 7.457.30 (m, 7 H), 6.85 (m, 2 H), 5.57-5.47 (m, 1 H), 5.23 (d, 1 H, J = 17.1 Hz), 5.12 (d, 1 H, J = 10.3 Hz), 4.95-4.91 (m, 1 H), 4.76 (dd, 1 H, J = 2.3, 7.8 Hz), 4.69 (t, 1 H, J = 8.7 Hz), 4.26-4.19 (m, 1 H), 4.06 (d, 1 H, J = 2.5 Hz), 3.78 (s, 3 H); the distinct signals of the cis isomer are: 5.75-5.67 (m, 1H), 5.5 (d, 1H, J = 17 Hz), 5.39 (d, 1H, J = 10.4 Hz).

Preparation of the compound of formula IX [(O) Reaction] 3-(4S-phenyl-oxazolidin-2-on-3-yl]-4-ethenyl-azetidinone 0.92 g of 1-Methoxyphenyl-3-[4S-phenyl-oxazolidin-2-on-3-yl]-4-ethenyl-azetidinone is dissolved in 70 ml of acetone and the solution is cooled in an ice/acetonitrile bath. 5.50 g of cerium ammonium nitrate dissolved in 70 ml of water is added dropwise. 30 minutes after the addition is complete, 100 ml of ethyl acetate is added to the mixture. The pH of the reaction mixture is increased to 7 by adding saturated sodium bicarbonate solution. The reaction mixture is stirred for 1 hour and then filtered through a filter bed containing Celite. The solution is diluted with ethyl acetate and washed with 10% sodium sulfite solution. (3x), saturated sodium chloride solution (1x), saturated sodium bicarbonate solution (2x), and finally saturated sodium chloride solution (1x). The organic solution is dried over sodium sulfate and concentrated on a rotary evaporator to give the desired 3-(4S-phenyloxazolidin-2-on-3-yl]-4-ethenylazetidinone β-lactam as a 4.6:1 trans:cis isomer mixture.

Trituration with ethyl acetate/hexane gave 0.47 g of a yellow solid.

TLC: R _f 0.16 (4/1 CHCl ₃ /ethyl acetate);

the spectrum data were obtained from a 4.3:1 trans:cis mixture, the integral values of the overlapping signals were corrected in some cases;

¹ H-NMR spectrum (CDCl, δ, 500 MHz): trans isomer 7.47-7.38 (m, 5 H), 6.24 (m, 1 H), 5.49-5.43 (m, 1 H), 5.15 (d, 1 H, J = 17.1 Hz), 5.02 (d, 1 H, J = 10.3 Hz), 4.94-4.87 (m, 1 H), 4.74 (t, 1 H, J = 8.8 Hz), 4.47 (d, 1 H, J = 2.5 Hz), 4.30-4.20 (m, 1 H), 3.88 (d, 1 H, J = 2.7 Hz); distinct signals of the cis isomer: 6.37 (m, 1 H), 5.85-5.78 (m, 1 H), 5.33-5.27 (m, 2 H), 4.69 (t, 1 H, J = 8.9 Hz).

Preparation of 3-[4S-phenyl-oxazolidin-2-on-3-yl]-4-[2-(1 naphthyl)ethen-1-yl-azetidinone [(P) Reaction]

0.1625 g of 1-Iodonaphthalene, 0.019 g of palladium acetate, 0.125 g of potassium acetate and 0.142 g of tetrabutylammonium chloride hydrate are dissolved in 1 ml of dimethylformamide. 0.11 g of 3-[4S-phenyloxazolidin-2-on-3yl]-4-ethenylazetidinone in 2 ml of dimethylformamide are added. The solution is heated at 80°C for 2.5 hours. After cooling to room temperature, the reaction mixture is filtered through a pad of Celite and diluted with ethyl acetate. The organic solution is washed with saturated sodium chloride solution (1x), saturated ammonium chloride solution (3x), saturated sodium chloride solution (1x), and saturated sodium bicarbonate solution (2x). The organic solution was dried over sodium sulfate and evaporated using a rotary evaporator to give a black oil. The product was purified on a silica filter with chloroform to remove the remaining 1-iodo-naphthalene, and the product was dissolved in ethyl acetate. Further purification by column chromatography (chloroform, then 1/1 chloroform/ethyl acetate, then chloroform) gave 0.062 g of [3-[4S-phenyl-oxazolidin-2-on-3-yl]-4-[2-(1-naphthyl)ethen-1-ylazetidinone β-lactam, as a 1:1 trans:cis isomer mixture.

TLC: R _f 0.36 (1/1 ethyl acetate/hexane);

the spectrum data were obtained from a 10:1 trans:cis mixture, the integral values of the overlapping signals were corrected in some cases;

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): trans isomer 7.997.96 (m, 1 H), 7.87-7.85 (m, 1 H), 7.81-7.79 (m, 1 H), 7.58-7.52 (m, 2 H), 7.43-7.23 (m, 7 H), 6.75 (s, 1H), 6.50 (s, 1H), 5.825.76 (m, 1H), 4.96-4.92 (m, 1H), 4.79-4.70 (m, 2H), 4.30-4.25 (m, 1H), 4.154.13 (m, 1H); the distinct signals of the cis isomer are: 8.10-8.07 (m, 1 H1, 6.55 (s, 1 H), 6.15-6.10 (m, 1 H).

Example 51

1-phenoxycarbonyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[2-(1-naphthyl)ethen-1-yl]azetidinone [P51/1 Reaction]

0.062 g of 3-[4S-Phenyl-oxazolidin-2-on-3-yl]-4-[2-(1-naphthyl)ethen-1-yl-azetidinone is dissolved in 3 ml of tetrahydrofuran and the solution is cooled with a dry ice/acetone bath. 0.15 ml of a 1 M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran is added. After 15 minutes, 22 μΙ of phenyl chloroformate in 1 ml of tetrahydrofuran is added. After 20 minutes, saturated ammonium chloride solution is added to the reaction mixture. Allow to warm to room temperature, then shake twice with chloroform and wash the organic part with saturated sodium chloride solution (1 x). The organic solution was dried over sodium sulfate and concentrated under reduced pressure to give 0.11 g of the desired β-lactam as a trans/cis isomeric mixture. 0.11 g of the desired trans β-lactam was

Obtained by column chromatography with 8/1 chloroform/ethyl acetate solvent.

TLC: R _f 0.42 (1/1 ethyl acetate/hexane);

H-NMR spectrum (CDCl3, 5, 500 MHz):8 7.94-7.91 (m, 1 H), 7.99-7.96 (m, 1 H) 7.82 (d, 1 H, J=88.2 Hz), 7.56-7.47 (m, 3 H), 7.45-7.34 (m, 9H), 7.32-7.23 (m, 1 H) 7.19 (d, 2H, J=7.9Hz), 5.90 (dd, 1 H, J=8.2, 15.4 Hz), 5.23 (dd, 1 H, J=3.5, 8.2 Hz), 5.05-5.02 (m, 1 H), 4.82 (t, 1 H, J= 8.7 Hz), 4.38-4.34 (m, 1H), 4.25 (d, 1H, J=3.7).

Preparation of 3-[4S-phenyl-oxazolidin-2-on-3-yl]-4-[(3-methoxyphenyl)ethen-1-yl]azetidinone [P51/2 Reaction]

0.203g of 3-Iodoanisole, 0.026g of palladium acetate, 0.171g of potassium acetate and 0.244g of tetrabutylammonium chloride hydrate are dissolved in 1 ml of dimethylformamide. 0.15g of 3[4S-phenyloxazolidin-2-on-3-yl]-4-ethenylazetidinone in 2 ml of dimethylformamide are added. The solution is stirred at 80°C for 3 hours. After cooling to room temperature, the reaction mixture is filtered through a filter pad containing Celite and washed with ethyl acetate. The organic solution is washed with saturated sodium chloride solution (1x), saturated ammonium chloride (3x), saturated sodium chloride solution (1x), and saturated sodium bicarbonate solution (3x). The organic solution was dried over sodium sulfate and concentrated on a rotary evaporator to give a black oil. The product was purified by column chromatography (chloroform, then 1/1 chloroform/ethyl acetate, then ethyl acetate) to give 0.068 g of the desired β-lactam [3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(3-methoxyphenyl)ethen1-yl]azetidinone.

Example 52

-phenoxycarbonyl-3-[4S-phenyloxazolidin-2-on-3-yl]-4-[2-(1-naphthyl)ethen-1-yl]azetidinone [Compound with formula P52]

In the process of Example 51, 0.007 g of the compound of formula P52 in cis configuration is obtained.

TLC: R _f 0.3 (1/1 ethyl acetate/hexane);

¹ H-NMR spectrum (CDCl3, 500 MHz): & 8.11 (d, 1 H, J=7.71 Hz), 7.96-7.80 (m, 2H), 7.657.19 (m, 15 H), 6.32 (dd, 1 H, J=8.6, 15.7 Hz) 5.08 (dd, 1 H, J=6.2, 8.5 Hz), 4.93 (t, 1 H, J = 8.1 Hz), 4.73-4.65 (m, 2 H), 4.27-4.24 (m, 1 H).

Example 53

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-methoxyphenyl)ethen-1-yl]azetidinone [P53/1 Reaction]

0.068 g of 3-[4S-Phenyl-oxazolidin-2-on-3-yl]-4-[3-methoxyphenyl)ethen-1-yl]-azetidinone was dissolved in 3 ml of tetrahydrofuran and the solution was cooled with a dry ice/acetone bath. 0.19 ml of a 1 M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran was added. After 15 min, 27 μΙ of phenyl chloroformate in 1 ml of tetrahydrofuran was added. After 20 min, saturated ammonium chloride solution was added to the reaction mixture. It was allowed to warm to room temperature, then shaken twice with chloroform and the organic portion was washed with saturated sodium chloride solution (1x). The organic solution was dried over sodium sulfate and evaporated under reduced pressure to give 0.082 g of the desired β-lactam. Purification by column chromatography with 9/1 chloroform/ethyl acetate gave 0.0321 g of product.

TLC: R _f 0.38 ((9/1 chloroform/ethyl acetate);

¹ H-NMR spectrum (CDCl3, δ, 500 MHz): 7.38 - 7.29 (m, 7H), 7.25 - 7.21 (m, 2H), 7.18 - 7.16 (m, 2H), 6.85 - 6.80 (m, 2H), 6.78 (s, 1Η), 6.42 (d, 1H, J = 15.8 Hz), 5.85 (dd, 1 H, J=7.7, 15.8 Hz), 5.05-5.03 (m, 1 Η), 5.00-4.97 (m, 1 II), 4.80 (t, 1 H, J=8.8 Hz), 4.37-4.34 (m, 1 H), 4.20 (d, 1 H, J=3.5 Hz), 3.82 (s, 3 H).

Preparation of 3-[4S-phenyl-oxazolidin-2-on-3-yl]-4-[(3-nitro-phenyl)ethen-1-yl]azetidinone [P53/2 Reaction]

0.217g of 3-Iodoanisole, 0.026g of palladium acetate, 0.171g of potassium acetate and 0.244g of tetrabutylammonium chloride hydrate were dissolved in 1 ml of dimethylformamide. 0.15g of 3-[4S-phenyloxazolidin-2-on-3-yl]-4-ethenylazetidinone in 2 ml of dimethylformamide were added. The solution was stirred at 80°C for 3 hours. After cooling to room temperature, the reaction mixture was filtered through a pad of Celite and washed with ethyl acetate. The organic solution was washed with saturated sodium chloride solution (1x), saturated ammonium chloride (3x), saturated sodium chloride solution (1x) and saturated sodium bicarbonate solution (3x). The organic solution was dried over sodium sulfate and concentrated on a rotary evaporator to give a black oil. The product was purified by column chromatography (chloroform, then 1/1 chloroform/ethyl acetate, then ethyl acetate) to give 0.033 g of the desired β-lactam [3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(3-nitrophenyl)ethen-1yl]azetidinone].

TLC: R _f 0.28 (1/1 CHCl ₃ /ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): 8.1 3-8.11 (m, 1 H), 8.02 (s, 1 H), 7.49 (d, 2 H, J = 5.1 Hz), 7.46-7.33 (m, 5 H), 6.45 (d, 1 H, J = 15.9 Hz), 6.17 (s, 1 H), 5.75 (dd, 1 H, J = 6.6, 15.9 Hz), 4.94-4.89 (m, 1 H), 4.80 (t, 1 H, J = 8.8 Hz), 4.61 (dd, 1 H, J = 2.2, 6.5 Hz), 4.39 (dd, 1 H, J = 5.5, 8.9 Hz), 3.98 (d, 1 H, J = 2.6 Hz).

Example 54

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3nitrophenyl)ethen-1-yl]azetidinone [P54 Reaction]

0.033 g of 3-[4S-phenyloxazolidin-2-on-3-yl]-4-[(3-nitrophenylethen-1-yl]azetidinone is dissolved in 3 ml of tetrahydrofuran and the solution is cooled with a dry ice/acetone bath. 0.1 ml of a 1 M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran is added. After 15 minutes, 12.7 μΙ of phenyl chloroformate in 1 ml of tetrahydrofuran is added. After 20 minutes, saturated ammonium chloride solution is added to the reaction mixture. Allow to warm to room temperature, then shake twice with chloroform and wash the organic part with saturated sodium chloride solution (1x). The organic solution is dried over sodium sulfate and evaporated under reduced pressure to obtain the desired β-lactam. With chloroform Triturated and washed with water, 0.007 g of product is obtained.

TLC: Rf 0.52 (4/1 chloroform/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): 8.12-8.09 (m, 2H), 7.72 (d, 1 H, J=7.7 Hz), 7.63 (t, 1 H, J=7.9 Hz), 7.46-7.42 (m, 4H), 7.34-7.28 (m, 3 H), 7.21-7.17 (m, 3 H), 6.66 (d, 1 H, J = 15.9 Hz) 6.30'(dd, 1 H, J=8.3, 15.9 Hz), 5.13 (dd, 1 H, J = 5.6, 8.6 Hz), 4.93 (dd, 1 H, J= 3.4, 8.3 Hz), 4.86 (t, 1 H, J=8.8 Hz), 4.73 (d, 1 H, J=3.6 Hz), 4.32 (dd, 1 H, J=5.6, 8.9 Hz).

55, Example

1-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-ethen-1-yl-azetidinone [Compound with formula P55]

0.16g of [3-[4S-Phenyl-oxazolidin-2-on-3-yl)-4-ethenyl-azetidinone is dissolved in 10 ml of tetrahydrofuran and the solution is cooled with a dry ice/acetone bath. 0.62 ml of a 1M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran is added. After 15 minutes, 90 μΙ of phenyl chloroformate in 1 ml of tetrahydrofuran is added. After 15 minutes, saturated ammonium chloride solution is added to the reaction mixture. Allow to warm to room temperature, then shake twice with chloroform and wash the organic portion with saturated sodium chloride solution (1x). The organic solution is dried over sodium sulfate and evaporated under reduced pressure to give the desired β-lactam. Column chromatography with 9/1 chloroform/ethyl acetate followed by recrystallization from hexane/ethyl acetate gave 0.07 g of product (31%) TLC: R _f 0.42 (9/1 chloroform/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz):8 7.49-7.43 (m, 3H), 7.40-7.36 (m, 4H), 7.29-7.24 (m, 1H), 7.20-7.18 (m, 2H), 5.65-5.57 (m, 1H), 5.22 (d, 1 H, J = l7.OHz), 5.17 (d, 1 H, J = 10.5 Hz), 4.98-4.92 (m, 2H), 4.80 (t, 1 H, J=8.8 Hz), 4.34 (dd, 1 H, J= 6.7, 8.8 Hz), 4.07 (d, 1 H, J=3.5 Hz).

The present invention also provides a new and advantageous process for the preparation of azetidinones, as well as intermediates useful in the preparation of azetidinones. More particularly, the invention relates to the individual reaction steps and the process for the preparation of a compound of formula Ia, wherein R ¹⁶ and R ¹⁷ are as previously defined;

according to which process, an aldehyde of general formula II, in which the group of general formula R ¹⁸ is a trialkylsilanyl group, and an amine of general formula III, protected at the nitrogen atom, in which the group of general formula R ¹⁹ is a nitrogen protecting group, are reacted to produce an imine of general formula IV, protected at the nitrogen atom, and then a nitrogen-protected azetidinone derivative of general formula VI is formed by reacting the imine of general formula IV, protected at the nitrogen atom, with 4S-phenyloxazolidin-2-on-3-ylacetyl chloride of formula V; by desilylation and epimerization of the nitrogen-protected azetidinone derivative of formula VI, a desilylated azetidinone intermediate of formula VII, substituted at the 4-position with an acetylene group, is obtained, by reduction of this intermediate and removal of the nitrogen-protecting group, the 3-(4S-phenyl-oxazolidin-2-on-3-yl)-4-ethenyl-azetidinone of formula IX' is obtained, in which the group of formula R ²⁰ is a tributylstannyl group or a group of formula B(OH) ₂ ; by reacting the compound of formula IX' in the presence of a catalyst with an electrophilic reagent of formula XR ¹⁶ (the group of formula X is a leaving group), and by acylation of the compound of formula IX', the substituted oxycarbonyl-azetidinone derivative of formula Ia is prepared.

The preferred intermediates of the present invention are characterized by the general formula VI, VII, VIII, IX', IXa and IXc, wherein the groups r ¹⁶ _R 17 _R 18 _{( r} i9. _{and r} 20) are as previously defined.

The following Reaction Scheme IV illustrates the preferred sequence of the reaction steps of this process, which is an object of the present invention. The groups R ¹⁶ , R ¹⁷ , R ¹⁸ , R ¹⁹ , and R ²⁰ in the Reaction Scheme are as previously defined.

By reducing the desilylated intermediate of general formula (VII) prepared according to Reaction Scheme II, substituted with an acetylene group at the 4-position, and removing the nitrogen protecting group, we obtain the trans3-(4S-phenyl-oxazolidin-2-on-3-yl)-4-[2-ethen-1-yl]azetidinone β-lactam of general formula (IX) [Reaction Steps 4 and 5]. It is advantageous to first prepare the trans-3-(4S-phenyl-oxazolidin-2-on-3-yl)-4-ethenyl-azetidinone β-lactam of formula (VIII) from the desilylated azetidinone intermediate of formula (VII) substituted with an acetylene group at the 4-position by removing the nitrogen protecting group, followed by partial reduction to form the 3-(4S-phenyl-oxazolidin-2-on-3-yl)-4-[2-ethen-1-yl]azetidinone of formula (IX). The removal of the protecting group is advantageously carried out, for example, as described in J. Orq. Chem., 47, 2765-68 (1982).

Partial reduction of the desilylated intermediate (VII) containing an acetylene group in the 4-position, or of the trans-3-(4S-phenyloxazolidin-2-on-3-yl)-4- ^{ethenylazetidinone} (VIII), can be carried out, for example, with an organic group-containing tin chloride, such as tributylone chloride, to give the desired tin-substituted vinyl derivative as a mixture of isomers. A catalyst can also be used, such as a palladium catalyst, such as dichlorobis(triphenylphosphine)palladium. Alternatively, the hydrostannylation of the triple bond can be carried out by thermolysis, for example in the presence of 2,2'-bisazoisobutyronitrile (AIBN). Other reducing agents, such as catecholborane or similar reagents, can give the vinylboronic acid-containing intermediate. The isomers can be separated by methods known per se, for example, by chromatography on silica gel, to give the desired 3-(4S-phenyl-oxazolidin-2-on-3-yl)-4-[2-ethen-1 yl]azetidinone of formula (IX'). It will be apparent to those skilled in the art that the partial reduction and deprotection can be carried out in any order.

The resulting trans-3-(4S-phenyl-oxazolidin-2-on-3-yl)-4[ethen-1-yl]-azetidinone [compound (IX')] is first functionalized by reacting with an electrophilic reagent of formula XR ¹⁶ (where X is a leaving group) in the presence of a solvent, and then the resulting compound is acylated to obtain the substituted oxycarbonyl-azetidinone derivative of formula Ia [Reaction Steps 6 and 7]. In formula XR ¹⁶ , X is a leaving group such as a halide, sulfonate, or the like, and R ¹⁶ is as defined above. The formation of the functional group is most advantageously carried out with the aid of a catalyst, such as tetrakis(triphenylphosphine)palladium(0), advantageously in tetrahydrofuran in the presence of copper(I) iodide. The following groups of general formula R are advantageous: 2-nitro-4-phenyl-, 4-nitro-2-phenyl-, 3-nitro-1-naphthyl-, 4-nitro-2naphthyl-, 2-quinolinyl-, 4-quinolinyl-, 7-quinolinyl-, 1-isoquinolinyl-, 3-isoquinolinyl-, 8-isoquinolinyl-, 2-benzothiazolyl-, 2-benzoxazolyl-. Even more advantageous R groups are: 2-thienyl-, 3-furanyl-, 5-pyrimidinyl-, and 3,5-dimethylphenyl-. It will be apparent to those skilled in the art that the functionalization of the trans-3-(4S-phenyloxazolidin-2-on-3-yl)-4-[2-ethen-1-yl]azetidinone of formula (IX') or the acylation of the compound can be carried out in any order, although the former reaction with the reagent of formula XR ¹⁶ , the product of which is the alkenyl-substituted azetidinone derivative of formula (XI'), advantageously precedes the acylation.

The acylation is advantageously carried out with a suitable chloroformate to give the substituted azetidinone derivative of formula Ia. Suitable chloroformates include phenyl chloroformate, 4-chlorophenyl chloroformate, 4-fluorophenyl chloroformate, benzyl chloroformate, 4-nitrobenzyl chloroformate, 4-chlorobutyl chloroformate, 4-methoxycarbonylphenyl chloroformate, 4-methoxyphenyl chloroformate, and allyl chloroformate.

Scheme IV illustrates the preferred sequence of reaction steps of the present invention. However, compounds of formula Ia may be prepared by altering the order of certain reaction steps. For example, compounds of formula Ia may be prepared by reducing the 4-acetylenically substituted azetidinone intermediate of formula VII, followed by deprotection to form trans-3-(4S-phenyloxazolidin-2-on-3-yl)-4-[2-ethen-1-yl]azetidinone of formula (IX'), followed by reaction with the electrophilic reagent of formula XR ¹⁶ and acylation (in either order). Alternatively, the 4-acetylenically substituted azetidinone intermediate of formula VII is first deprotected and then reduced to form trans-3-(4S-phenyloxazolidin-2 _-one -3-yl)-4-[2-ethen-1-yl]azetidinone of formula (IX'), which is reacted with the electrophilic reagent of formula XR ¹⁶ , followed by deprotection and acylation. Alternatively, the 4-acetylenically substituted azetidinone intermediate of formula VII is first deprotected and then reduced to form 3-(4S-phenyloxazolidin-2-one-3-yl)-4-[2-ethen-1-yl]azetidinone of formula IX', followed by reaction with the electrophilic reagent of formula XR ¹⁶ , followed by acylation (in any order).

The following Preparations and Examples further illustrate the preparation of the compounds of the present invention and are not intended to limit the scope of the invention in any way.

Preparation of the compound of formula VIII [(Q) Reaction]

2.0 g (5.5 mmol) of the azetidinone derivative (Vila) was dissolved in 140 ml of acetonitrile and the solution was cooled to -15°C in an ice/acetone bath. 121.1 g (22.0 mmol) of cerium ammonium nitrate in 140 ml of water was added dropwise. Then 200 ml of ethyl acetate was added. The pH of the reaction mixture was adjusted to 7 with saturated sodium bicarbonate solution. After stirring for 1 hour, the reaction mixture was filtered under reduced pressure through a pad of Celite. The organic layer was separated and the aqueous layer was extracted with ethyl acetate. The combined organic phases were washed with 10% sodium sulfite solution, saturated sodium chloride solution, and saturated sodium bicarbonate solution. The organic solution was dried over anhydrous sodium sulfate and concentrated in a rotary evaporator. The yellow solid was triturated with ethyl acetate to give 0.55 g of compound (VIII). The residue was purified by column chromatography to give an additional 0.25 g of product, total yield: 0.80 g.

TLC: R _f 0.20 (4/1 chloroform/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): 7.49-7.44 (m, 3 H), 7.37-7.35 (m, 2 H), 6.05 (s, 1 H), 4.96 (dd, J = 6.9, 8.7 Hz, 1 H), 4.76 (t, J = 8.9 Hz, 1 H), 4.69 (t, J = 2.3 Hz, 1 H), 4.28 (m, 2 H), 2.29 (d, J = 2.1 Hz, 1 H).

Preparation of compounds of formula IXa and IXb [(R) Reaction]

1.78 g (7.0 mmol) of compound VIII was dissolved in 70 ml of tetrahydrofuran. 0.10 g (0.14 mmol) of dichlorobis(triphenylphosphine)palladium was added and the mixture was stirred under nitrogen. 2.3 ml (8.4 mmol) of tributyltin was then added dropwise. At the end of the addition, hydrogen evolution and slight warming were observed in the reaction mixture. After the addition was complete, the solvent was evaporated to give an orange oil. Column chromatography (3/1 -> 1/1 hexane/ethyl acetate) gave 2.54 g (66%) of the desired tin-containing intermediate (IXa), contaminated with about 7% of the tin derivative substituted at the α-position. The desired tin derivative of formula (IXa) is used in the next step without further purification.

TLC: R _f 0.38 (1/1 hexane/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): 7.40-7.33 (m, 5 H), 6.32 (s, 1 H), 6.12 (m, 1 H), 5.53 (m, 1 H), 4.91 (dd, J = 6.3, 8.6 Hz, 1 H), 4.73 (t, J = 8.8 Hz, 1 H), 4.43 (m, 1 H), 4.24 (dd, J = 6.2, 8.8 Hz, 1 H), 3.84 (d, J = 2.5 Hz, 1 H), 1.41 (m, 6 H), 1.27 (m, 6 H), 0.85 (m, 15 H).

Example 56 [ΓΡ56/1 Reaction]

0.198 g (0.362 mmol) of the tin intermediate IXa was dissolved in 5 ml of deoxygenated tetrahydrofuran (deoxygenation was performed by flushing the solvent vapor space with nitrogen for 1 h). To the solution were added 0.052 ml (0.471 mmol) of 2-iodothiophene, 0.042 g (0.036 mmol) of tetrakis(triphenylphosphine)palladium(0) and 0.0138 g (0.0724 mmol) of copper iodide. The reaction mixture was first stirred at room temperature for 1.25 h and then boiled under a nitrogen blanket for 1.5 h. 200 ml of ethyl acetate were added and the solution was washed with saturated sodium chloride solution, saturated sodium bicarbonate solution and water. The organic solution was dried over anhydrous sodium sulfate and concentrated by rotary evaporation. The residue was dissolved in acetonitrile and washed with hexane. Evaporation gave 0.17 g of a yellow solid which was purified by column chromatography to give 0.06 g of the desired product. TLC: R _f 0.24 (3/2 chloroform/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): 7.41-7.32 (m, 5 H), 7.17 (d, J = 5.0 Hz, 1 H), 6.96 (m, 1 H), 6.89 (d, J = 3.4 Hz, 1 H), 6.52 (d, J = 15.7 Hz, 1 H), 6.23 (s, 1 H), 5.50 (dd, J = 7.1, 15.7 Hz, 1 H), 4.90 (dd, J = 5.8, 8.6 Hz, 1 H), 4.76 (t, J = 8.8 Hz, 1 H), 4.54 (dd, J = 2.3, 7.1 Hz, 1 H), 4.33 (dd, J = 5.8, 8.9 Hz, 1 H), 3.96 (d, J=2.6 Hz, 1H).

-phenoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(2thienyl)ethen-1-yl]azetidinyl [P56/2 Reaction]

0.06 g (0.176 mmol) of the starting azetidinone derivative [P56/2 Reaction] was dissolved in 5 ml of tetrahydrofuran and the solution was cooled with a dry ice/acetone bath. 0.181 (0.18 mmol) of a 1 M solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran was added. After 15 min, 0.024 ml (0.15 mmol) of phenyl chloroformate in 1 ml of tetrahydrofuran was added. After 30 min, saturated ammonium chloride solution was added to the reaction mixture. Allow to warm to room temperature, then shake twice with chloroform and wash the organic portion with saturated sodium chloride solution (1x). The organic solution was dried over anhydrous sodium sulfate and concentrated under reduced pressure to give 0.09 g of a yellow solid. Column chromatography with 9/1 chloroform/ethyl acetate gave 0.055 g (68%) of trans β-lactam, contaminated with approximately 7% of cis β-lactam.

TLC: Rf 0.38 (9/1 chloroform/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): 7.33 (m, 11 H), 6.96 (m, 1 H), 6.90 (d, J = 3.3 Hz, 1 H), 6.53 (d, J = 15.6 Hz, 1 H), 5.65 (dd, J = 7.8, 15.6 Hz, 1 H), 4.97 (m, 2 H), 4.81 (t, J = 8.8 Hz, 1 H), 4.37 (dd, J = 6.3, 8.9 Hz, 1 H), 4.16 (d, J = 3.5 Hz, 1 H).

Example 57 [ΓΡ57/1 Reaction]

The product was prepared from 0.20 g (0.37 mmol) of the tin intermediate IXa by the procedure of Example 7A with 0.042 ml (0.475 mmol) of 3-bromofuran. Instead of tetrakis-(triphenylphosphine)palladium(0), 10 mol% tris(dibenzylideneacetone)dipalladium and 80 mol% tri-2-furylphosphine were used. The reaction mixture was first stirred at room temperature for 1 hour and then refluxed for 16 hours. After work-up, the material obtained was filtered through a filter pad containing silica gel (1/1 chloroform/ethyl acetate, then ethyl acetate). The collected product was dissolved in acetonitrile and washed with hexane. Evaporation gave 14 mg of a yellow solid.

TLC: Rf 0.23 (ethyl acetate);

¹ H-NMR spectrum (CDCl ₃ , δ, 500 MHz); 7.70 (m, 1 H), 7.35 (m, 6 H), 7.00 (m, 1 H), 6.55 (m, 1 H), 6.25 (m, 1 H), 5.81 (m, 1 H), 5.17 (m, 1 H), 4.88 (m, 1 H), 4.42 (m, 1 H), 4.25 (m, 1H), 3.88 (m, 1H).

-phenoxycarbonyl-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-furanyl)ethen-1-yl]azetidinone [P57/2 Reaction]

0.20 g (0.37 mmol) of the starting azetidinone derivative [P57/2 Reaction] was dissolved in 1 ml of methylene chloride. A catalytic amount of 4-dimethylaminopyridine was added, followed by 0.06 ml (0.43 mmol) of triethylamine. After 5 min, 0.022 ml (0.17 mmol) of phenyl chloroformate was added. After 30 min, chloroform was added to the solution and the mixture was washed with saturated ammonium chloride solution (3x) and then with saturated sodium chloride solution (1x). The organic solution was dried over anhydrous sodium sulfate and evaporated under reduced pressure to give 0.039 g of an orange solid. Triturated in hexane/ethyl acetate, then silica gel was added.

Filtration on a filter bed containing Kagel with a 1/2 hexane-ethyl acetate solvent mixture gave the desired compound (5.5 mg).

TLC: R _f 0.27 (2/1 chloroform/ethyl acetate);

1H-NMR spectrum (CDCl3, 500 MHz): 8 7.31 (m, 13 H), 5.87 (d, J = 14.1 Hz, 1 H), 5.33 (m, 1 H), 4.93 (m, 2 H), 4.79 (t, J = 8.8 Hz, 1 H), 4.35 (dd, J = 6.6, 8.5 Hz, 1 H), 4.07 (d, J = 3.3 Hz, 1 H).

Example 58 [ΓΡ58/1 Reaction]

The product of Reaction P58/1 was prepared from 0.20 g (0.55 mmol) of the tin intermediate IXa according to the procedure of Example 7A with 0.11 g (0.71 mmol) of 5-bromopyridine. Instead of tetrakis-(triphenylphosphine)palladium(0), 10 mol% tris(dibenzylideneacetone)dipalladium and 80 mol% triphenylrilarzine were used. The reaction mixture was refluxed for 4 hours. The resulting material was dissolved in ethyl acetate and washed with 1 N hydrochloric acid solution (4x). The pH of the aqueous portions was adjusted to 6 with 5 N sodium hydroxide solution and then to 8.5 with saturated sodium bicarbonate solution. The product was extracted from the basic portion with ethyl acetate (3x). The organic solution was dried over anhydrous sodium sulfate and concentrated on a rotary evaporator to give 28 g of the desired product as a white solid.

TLC: R _f 0.23 (ethyl acetate);

¹ H-NMR spectrum (CDCl ₃ , δ, 500 MHz): 9.09 (s, 1 H), 8.55 (s, 2 H), 7.35 (m, 5 H), 6.54 (s, 1 H), 6.31 (d, J = 16.1 Hz, 1 H),

5.80 (dd, J = 6.5, 16.0 Hz, 1H), 4.90 (dd, J = 5.4, 8.7 Hz, 1H),

4.79 (t, J = 8.9 Hz, 1 H), 4.59 (dd, J = 2.1, 6.6 Hz, 1 H), 4.37 (dd, J = 5.3, 8.9 Hz, 1 H), 3.98 (d, J = 2.7 Hz, 1 H).

-phenoxycarbonyl-3 R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(5-pyrimidinyl)ethen-1-yl]azetidinone [P58/2 Reaction]

The product of Reaction P58/2 was prepared from 28 mg of the azetidinone derivative (ix) by the procedure of Example 8B. Recrystallization from diethyl ether/ethyl acetate gave 18 mg of the desired compound.

TLC: R _f 0.52 (ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , Ö, 500 MHz): 9.15 (s, 1 H), 8.59 (s, 2 H), 7.35 (m, 8 H), 7.19 (d, J = 7.9 Hz, 1 H), 6.33 (d, J = 16.1

Hz, 1 H), 5.98 (dd, J = 7.5, 16.1 Hz, 1 H), 5.02 (dd, J = 3.5, 7.4

Hz, 1 H), 4.95 (dd, J = 5.8, 8.5 Hz, 1 H), 4.84 (t, J = 8.8 Hz, 1

H), 4.43 (dd, J = 5.8, 8.9 Hz, 1 H), 4.18 (d, J = 3.6 Hz, 1 H).

Example 59 [P59/1 Reaction]

The product of Reaction P58/1 was prepared from 0.20 g (0.37 mmol) of the tin intermediate IXa by the procedure of Example 7A with 0.11 g (0.48 mmol) of 5-iodo-m-xylene. Instead of tetrakis(triphenylphosphine)palladium(0), 10 mol% tris(dibenzylideneacetone)dipalladium and 80 mol% triphenylarsine were used. The reaction mixture was refluxed for 1.5 hours. The material obtained after workup was purified on a filter bed containing silica gel with a 1/1 chloroform/ethyl acetate solvent. The collected product was dissolved in acetonitrile and washed with hexane (4x). The 0.044 g of yellow material obtained by evaporation was further purified by radial chromatography (1/1 chloroform/ethyl acetate solvent mixture) to give a product with a purity of more than 90% according to the NMR spectrum.

TLC: R _f 0.20 (1/1 hexane/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , δ, 500 MHz): 7.35 (m, 5 H), 6.91 (s, 1 H), 6.82 (s, 2 H), 6.45 (s, 1 H), 6.35 (d, J = 15.8 Hz, 1 H), 5.66 (dd, J = 7.1, 15.8 Hz, 1 H), 4.90 (dd, J = 6.0, 8.5 Hz, 1 H), 4.75 (t, J = 8.8 Hz, 1 H), 4.57 (dd, J = 1.6, 6.9 Hz, 1 H), 4.29 (dd, J = 6.0, 8.8 Hz, 1 H), 3.98 (d, J = 2.4 Hz, 1 H).

-phenoxycarbonyl-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3,5-

-dimethyl-phenyl)ethen-1-yl]azetidinone [P59/2 Reaction]

The product of Reaction P59/2 was prepared from 0.044 g of the product of Reaction P59/1, according to the procedure of Example 8B. Recrystallization from diethyl ether/ethyl acetate gave 0.032 g of the desired compound.

TLC: R _f 0.65 (1/1 hexane/ethyl acetate);

¹ H-NMR spectrum (CDCI ₃ , Ö, 500 MHz): 7.38 (m, 7 H), 7.24 (m, 1 H), 7.18 (d, J = 7.9 Hz, 2 H), 6.93 (s, 1 H), 6.84 (s, 2 H), 6.43 (d, J = 15.8 Hz, 1 H), 5.85 (dd, J = 7.7, 15.8 Hz, 1 H), 5.07 (dd, J = 3.6, 7.6 Hz, 1 H), 4.98 (dd, J = 6.7, 8.5 Hz, 1 H), 4.81 (t, J = 8.8 Hz, 1 H), 4.36 (dd, J = 6.7, 8.8 Hz, 1 H), 4.16 (d, J = 3.6 Hz, 1 H).

Example 60: trans-z-1-isopropyloxycarbonyl-3R-F4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [P60 Reaction]

0.1 g (0.3003 mmol) of the unprotected trans azetidinone derivative of formula (vi) was dissolved in 5 ml of dichloromethane at room temperature. 0.084 ml (0.6006 mmol) of triethylamine and a catalytic amount of DMAP were added in one portion. 0.6 ml of a 1.0 M solution of isopropyl chloroformate in toluene was added dropwise to the reaction mixture at room temperature. The resulting solution was stirred at room temperature overnight and then purified on silica gel to give 73 mg of product.

Knee desorption MS = 420.9

100

Analysis

Calculated (%): C 65.55 H 5.50 N 9.97

Found (%): C 65.67 H 5.36 N 9.86

The compounds of Examples 61-73 were prepared by the procedure of Example 60 using 5 equivalents of the corresponding chloroform.

Example 61: trans-1-sec-butoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone

[Compound with formula P61]

Yield: 41 mg of white solid: MS=435

Example 62. Trans-1-D-nitrobenzyloxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone

[Compound with formula P62]

Yield: 75 mg of product, almost white solid: MS =

Analysis

Calcd (%): C 58.56 H 4.21 N 10.17

Found (%): C 59.09 H 3.96 N 10.14

Example 63: trans-1-ethoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [Compound of Formula P63]

Yield: 52 mg product; MS = 406

IOI

64, Example trans-1-[2-(2-isopropyl-5-methyl)-cyclohexyloxycarbonyl]-3R-[4-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone

[Compound with formula P64]

Yield: 34 mg of product; MS = 517.6

Example 65: trans-1-methoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R(2-(3-pyridyl)ethen-1-yl)azetidinone [Compound of Formula P65]

Yield: 61 mg of white solid; MS = 393.1

Example 66: trans-1-benzyloxycarbonyl-3-R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [Compound of Formula P66]

Yield: 69 mg solid; MS = 469

Example 67: trans-1-cyclohexyloxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [Compound of formula P67]

Yield: 24 mg solid; MS=461.5

Example 68: trans-1-cyclobutoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [Compound of Formula P68]

Yield: 31 mg of white solid; MS=433.4

Example 69: trans-1-cyclopentyloxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [Compound of formula P69] Yield: 27 mg of foam; MS = 447.5

102

Example 70: trans-1-allyloxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [Compound P70] Yield: 63 mg of white foam; MS = 419

Example 71: trans-1-n-butoxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (compound P71) Yield: 71 mg of yellow solid; MS=435.2

Example 72: trans-1-isobutoxycarbonyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [Compound P72] Yield: 50 mg of white solid; MS=435.2

Example 73: trans-1-vinyloxycarbonyl-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2(3-pyridyl)ethen-1-yl]azetidinone [Compound P73] Yield: 17 mg solid; MS=405.4

Example 74 [P74 Reaction] trans-1-[benzoxazol-2-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (vii)]

To a solution of 0.68 g (6.7 mmol) of the unprotected azetidinone derivative (vi) in 4 ml of dichloromethane were added 0.668 g (6.7 mmol) of triethylamine, a catalytic amount of dimethylaminopyridine (DMAP), and 0.21 g of 2-chlorobenzoxazole. The resulting reaction mixture was stirred for 5 hours and then diluted with ethyl acetate. The organic solution was extracted first with 0.5 N aqueous hydrochloric acid solution and then with 1 N aqueous hydrochloric acid solution.

103 zuk. The aqueous portions were combined and the pH of the solution was adjusted to approximately 7 by addition of saturated sodium bicarbonate solution. The mixture was extracted with ethyl acetate, the organic solution was dried over sodium sulfate, filtered, and concentrated by rotary evaporation. The residue was purified by chromatography on silica gel (ethyl acetate as solvent). The product was evaporated from chloroform twice and dried at 40°C to give 0.11 g of compound (vii) (34% yield).

The spectral data were collected from two similarly prepared samples containing approximately 0.5 equivalents of water.

TLC: _Rf (EtOAc) 0.44;

¹ H-NMR spectrum (CDCl ₃ , δ, 300 MHz); 8.5 (br s, 1H), 8.4 (br s, 1H), 7.57-7.21 (m, 11H), 6.45 (d, 1H, J = 15.9Hz), 5.94 (dd, 1H, J=7.5, 15.9Hz), 5.19 (dd, 1H, J=3.15, 7.5Hz), 4.97 (dd, 1H, J = 5.8, 8.6 Hz), 4.82 (t, 1H, J = 8.8 Hz), 4.40 (dd, 1H, J = 5.8, 8.8 Hz), 4.35 (d, 1H, J = 3.3 Hz);

IR spectrum (CHCl ₃ ): 1799, 1760, 1624, 1574 cm'¹;

MS (FD+) 452;

Analysis based on the formula C ₁₇ H ₁₅ N0 ₅ *1/2H ₂ 0

Calcd (%): C 67.67 H 4.59 N 12.14

Found (%): C 67.88 H 4.57 N 11.96

Example 75 [P75 Reaction] trans-1-[4,6-dimethoxy-1,3,5-triazin-2-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (viii)]

0.18 g (0.53 mmol) of the unprotected azetidinone derivative of formula (vi) and 0.19 g (1.1 mmol) of 2-chloro

104

To a solution of 4,6-dimethoxy-1,3,5-triaziη in 4 ml of dichloromethane was added 0.34 g (2.63 mmol) of Ν,Ν-diisopropylethylamine and a catalytic amount of dimethylaminopyridine (DMAP). The reaction mixture was stirred under a nitrogen blanket for 3 h, then diluted with dichloromethane and washed with saturated ammonium chloride solution. The organic layer was dried over sodium sulfate, filtered and evaporated. The residue was purified by chromatography on silica gel (gradient: 100% hexane -> 75% hexane containing ethyl acetate) to give 0.04 g of the desired compound of formula (viii) (16% yield).

TLC: _Rf (EtOAc) 0.26;

¹ H-NMR spectrum (CDCI ₃ , δ, 300 MHz): 8.4 (br s, 2H), 7.5-7.3 (m, 7H), 6.31 (d, 1H, J = 1 5.9 Hz), 5.90 (dd, 1H, J=7.2, 15.9 Hz), 5.08 (dd, 1H, J=2.5, 6.8 Hz), 4.95 (m, 1H), 4.80 (t, 1H, J=8.6 Hz), 4.37 (dd, 1H, J = 5.4, 8.6 Hz), 4.17 (d, 1H, J=3.1 Hz), 3.96 (s, 6H).

The compounds of Examples 76, 77, and 78 (trans-1-[4-trifluoromethyl-2-pyrimidyl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (ix)]; trans-1-[6-nitro-pyridin-2-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (x)]; trans-1-[2-phenyl-quinazol-4-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (xi)]) were prepared by adding 100 mg. (0.3003 mmol) of the unprotected azetidinone derivative of formula (vi) is dissolved in 5 ml of dichloromethane at room temperature. 0.21 ml (4.5 mmol) of triethylamine and a catalytic amount of DMAP are added in one portion and the reaction mixture is stirred. The appropriate chlorinated heterocyclic compound is added to the reaction mixture while stirring. ·..···:·...··..· .:.

letet (2-chloro-4-(trifluoromethyl)pyrimidine; 2-chloro-5-nitropyridine;

or 4-chloro-2-phenylquinazone) and stirred at room temperature overnight. Purification of the resulting residue on silica gel affords the desired product.

Example 76: trans-1-[4-trifluoromethyl-2-diimidyl-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (ix)]

Yield: 27 mg; MS=481.1

77. Example trans-1-[6-nitro-pyridin-2-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]4R-[2-(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (x)] Yield; 33 mg; MS=457.1

78. Example trans-1-[2-phenylquinazol-4-yl-3R-[4S-phenyloxazolidin-2-on-3-yl]14R-[2-(3-pyridyl)ethen-1-yl]azetidinone [compound of formula (xi)]

Yield: 18 mg; MS=539

Example 79: trans-1-[benzoxazol-2-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yl]azetidinone [Compound of formula P79]

0.134 g (0.40 mmol) of compound (vi) was dissolved in 10 ml of dichloromethane. 0.2 g of diisopropylethylamine was added, followed by 2-chlorobenzoxazole. The resulting reaction mixture was stirred overnight, then 0.098 g (0.8 mmol) of dimethylaminopyridine was added and stirring was continued for a further 4 hours. The reaction mixture was diluted with ethyl acetate and washed with saturated sodium chloride solution. The organic layer was dried over sodium sulfate, filtered and evaporated to give an oil. The residue was purified by chromatography on silica gel (1:1 hexane:ethyl/acetate). Further purification was carried out by recrystallization from acetonitrile to give 20 mg of the desired product.

¹ H-NMR (CDCI ₃ , δ, 300 MHz): 7.59 - 7.19 (m, 14 H), 6.54 (d, 1H, J = 15.9Hz), 5.88 (dd, 1H, J=7.7, J = 15.8 Hz), 5.22 (dd, 1H, J = 3.0, J = 7.6 Hz), 4.96 (dd, 1H, J=6.3, J=8.6 Hz), 4.80 (t, 1H, J=8.8Hz), 4.36 (dd, 1H, J=6.3, J = 8.9 Hz), 4.32 (d, 1H, J=3.3 Hz).

Example 80: cis-1-[benzoxazol-2-yl]-3R-[4S-phenyl-oxazolidin-2-on-3-yl]-4R-[2-(phenyl)ethen-1-yl]azetidinone [Compound P80]

0.4 g (1.2 mmol) of the unprotected 3,4-cis-plactam isomer was dissolved in 4 ml of dichloromethane and 1.2 g (11 mmol) of triethylamine was added, followed by 0.36 g (2.4 mmol) of 2-chlorobenzoxazole and about 0.05 g (0.4 mmol) of dimethylaminopyridine. The resulting reaction mixture was stirred overnight. The reaction mixture was diluted with ether and washed first with 0.5 N hydrochloric acid, then with saturated sodium bicarbonate solution and finally with saturated sodium chloride solution. The organic phase was dried over sodium sulfate, filtered and evaporated to give an oil. The residue was purified by chromatography on silica gel (1:1 hexane:ethyl acetate).

TLC: R _f (1:1 hexane:ethyl acetate) 0.64;

¹ H-NMR spectrum (CDCI ₃ , δ, 300 MHz): 7.56-7.20 (m, 14H), 6.90 (d, 1H, J = 16Hz), 6.31 (dd, 1H, J=8.6, J = 16Hz), 5.10 (dd, 1H, J=5.8, J = 8.7Hz), 4.90 (t, 1H, J = 8.4 Hz), 4.76 (d, 1H, J = 5.7 Hz), 4.68 (t, 1H, J = 8.9 Hz), 4.23 (t, 1H, J = 8.0 Hz).

The thioamido-substituted azetidinone derivatives of the present invention are prepared by various methods known per se. Scheme V (wherein R ²³ is as defined below) shows a preferred method for preparing certain preferred thioamido-azetidinone derivatives of formula Ib, and their pharmaceutically acceptable salts or solvates, wherein R ²³ is phenyl, benzyl, naphthyl, or furfuryl; and the aromatic ring of the phenyl, naphthyl, or benzyl may be substituted with one to four halogen, trifluoromethyl, or C 1-6 alkyl groups;

To prepare the compound of formula Ib, 250 mg (0.7462 mmol) of the unprotected trans azetidinone derivative (vi) was dissolved in dry tetrahydrofuran. The reaction mixture was cooled to about -78°C using an ice/acetone bath, and then 125 mg (0.7462 mmol) of lithium bis(triethylsilyl)amide in 0.75 mL of 1 M tetrahydrofuran was slowly added. The reaction mixture was stirred for 30 minutes, then the appropriate isothiocyanate (2 equivalents) was added and stirred for 30 minutes. The appropriate isothiocyanate was commercially available or could be prepared by methods known per se. The reaction mixture was allowed to warm to room temperature and then purified on silica gel to give the compound of formula Ie.

The following Examples are intended to further illustrate the preparation of the compounds of the present invention and are not intended to limit the scope of the invention.

iOS ’ ’ * '

Example 81 [P81 Reaction] trans-1-phenylthioamido-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[9(3-pyridyl)ethen-1-yl]azetidinone

250 g (0.7462 mmol) of the deprotected trans-azetidinone derivative (vi) was dissolved in 125 ml of dry tetrahydrofuran and cooled to about -78°C using an ice/acetone bath. 125 mg (0.7462 mmol) of lithium bis(triethylsilyl)amide in 0.75 ml of 1 M tetrahydrofuran was slowly added to the reaction mixture. The reaction mixture was stirred for 30 min, then 263 mg of phenylisothiocyanate was added and stirred for 30 min. The resulting solution was stirred overnight, allowed to warm to room temperature, and purified on silica gel to give the desired product. MS = 470

The compounds of Examples 82-89 were prepared by the method of Example 81 using 2 equivalents of the appropriate isothiocyanate.

Example 82: trans-1-[2-bromophenylthioamidol-3R-[4S-phenyloxazolidin-2-on-3yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (2-bromophenyl isothiocyanate) [Compound P82]

Yield: 266 mg of white solid: MS=549

Example 83. Trans-1-[3-fluorophenylthioamidol-3R-4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (3-fluorophenyl isothiocyanate) [Compound of formula P83]

Yield: 210 mg of white solid: MS=488

109

Example 84. Trans-[2-fluoro-phenylthioamidol-3R-[4S-phenyl-oxazolidin-2-on-3yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (2-fluoro-phenyl isothiocyanate) [Compound of formula P84]

Yield: 100 mg product; MS=488

Example 85: trans-1-[4-fluorophenylthioamidoloxycarbonyl-3R-(4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (4-fluorophenyl isothiocyanate) [Compound P85]

Yield: 40 mg of black solid; MS=488

Example 86: trans-1-[2,3,5,6-tetrafluorophenylthioamidol-3R-(4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (2,3,5,6-tetrafluorophenyl isothiocyanate) [Compound P86] Yield: 20 mg of white solid; MS=541.8

Example 87: trans-1-[3-furfurylthioamido-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (3-furfuryl isothiocyanate) [Compound P87]

Yield: 70 mg of black solid; MS=473.9

Example 88. trans-1-naphthylthioamido-3R-[4S-phenyloxazolidin-2-on-3-yl]-4R-[2(3-pyridyl)ethen-1-yl]azetidinone (2-naphthyl isothiocyanate)

[Compound with formula P88]

Yield: 55 mg of white solid; MS=520

10

Example 89: trans-1-[4-trifluoromethylphenylthioamido]-3R-(4S-phenyloxazolidin-2-on-3-yl]-4R-[2-(3-pyridyl)ethen-1-yl]azetidinone (4-trifluoromethylphenyl isothiocyanate) [Compound P89]

Yield: 30 mg of yellow, foamy substance; MS=538.0

The biological activity of the compounds of the present invention was evaluated by a preliminary screening assay that measures the inhibitory effect of the test compound on the proteolytic activity of PSA.

Colorimetric method for measuring PSA inhibitory activity

The IC ₅₀ value (concentration causing 50% inhibition) of the compounds as PSA inhibitors was determined after 2 hours of preincubation in the presence of PSA [prostate-specific antigen - protein concentration 0.72 mg/ml (25.3 prnol) stored at -20°C in 50% glycerol PBS (phosphate buffered saline); molecular weight = 28.433 g/mol; the specific activity of PSA was experimentally determined to be 155 mmol/hr/mg protein according to the Beer-Lambert law]. Two different PBS/BSA buffer solutions are used (BSA = bovine serum albumin): Buffer solution “A”, consisting of PBS + 0.7 mg BSA/ml, is used to dilute PSA, and Buffer solution “B”, consisting of PBS + 6.3 mg BSA/ml, is used in the reaction mixture to prevent non-specific binding of PSA to the microtiter plates. Both buffer solutions are stored at 4°C. Disposable, non-sterile, polystyrene (untreated), 96-well, flat-bottomed microtiter plates from Corning are used (catalog number: #25880-96). A Bio-Tek Instruments Inc. CERES UV900HDI microtiter plate reader is used to read the colorimetric changes. To estimate optical density changes, readings are taken at two wavelengths: 405 nm and a reference wavelength of 450 nm.

The peptide substrate (MeO-Suc-Arg-Pro-Tyr-pNA) was obtained from Pharmacia Hepar [Chromogenix S-2586 (Chymotrypsin Chromogenic Substrate; Catalog No. S-2586 (25 mg tubes); molecular weight = 705.3; store at room temperature (dry) until reconstitution; reconstitute in PBS at 10 mM (7.05 mg/ml) and store at 4°C as described in the product information]. PBS was Gibco BRL 1X phosphate buffered saline (D-PBS, Catalog No. 14040-026), which was stored at 4°C. BSA (Bovine Serum Albumin) was obtained from Sigma (Cat. No. A-7511) and stored at 4°C. we store it.

The PBS/BSA solutions and the substrate solution (3546 ml PBS/25 mg substrate) are removed from the refrigerator and kept at 37°C for at least 30 minutes. 1-5 mg of the material to be tested for PSA inhibition (solid or oil) are weighed and diluted with DMSO to prepare a 300 pmol inhibitor stock solution. Dilution series of the inhibitor stock solution are prepared in a 96-well microtiter plate with DMSO solvent, so that the final inhibitor concentrations in the reaction mixture are as follows: 100; 33; 10; 3; 1; 0.3; 0.1; 0.03; and 0.01 pmol. The outer wells of the plates are not used. Rows A - H and columns 1 - 12 are used on the plate. Hole B2 served as a blank in all experiments.

Add 10 μΙ of PBS Buffer Solution “B” to all reaction wells, including B2. Add 10 μΙ of DMSO to well B2 and to the “control without inhibitor” wells (B10, C10, D10, E10, F10, G10, H10).

Add 10 μΙ of the sample to be tested to each of the other wells (B1-9, C1-9, D1-9, E1-9, F1-9, G1-9 and H1-9).

μΙ PSA stock solution is diluted with 610 μΙ PBS Buffer Solution “A”. 1ΟμΙ PBS + 0.7 mg BSA/ml solution is added to wells B2. 10μΙ diluted PSA solution is added to the wells containing the inhibitor and to the “control wells without inhibitor” (40 nmol final PSA concentration). The microtiter plate is gently shaken by hand to mix the contents of the wells properly, the wells are sealed with parafilm and pretreated for 2 hours at 37°C. After pretreatment, 70μΙ substrate stock solution is added to each well (1 mmol final substrate concentration). The reaction progress is monitored for the next 10 minutes, i.e. the cleavage of the β,-nitroaniline (β-ΝΑ) moiety from the substrate by PSA, which results in the appearance of a yellow color. The reader reads twenty optical density values per well, which are stored in the device.

After data collection is complete, a printed curve of the reaction progress is obtained for each well. The reaction rates (slope of the curves in [thousandths of optical density units]/min) are plotted against inhibitor concentration to determine the concentration causing 50% inhibition (IC ₅₀ ).

IC ₅₀ values are determined by non-linear curve analysis. Compounds for which the inhibitory effect at the highest concentration (usually 10ΟμΓπόΙ/Ι) did not reach 50% are assigned an IC ₅₀ value (> the highest concentration). For compounds with an inhibitory effect greater than 50% at the highest inhibitor concentration, the IC ₅₀ value is determined by non-linear analysis.

Results of the colorimetric measurement:

Example# IC50 pmol/l Example# IC50 pmol/l Example# IC50 μπΊόΙ/Ι 1 0.20 31 0.34 60 11 2 0.24 32 0.95 61 20.32 3 0.30 33 13.03 62 27.95 4 0.69 34 0.13 63 5.4 5 0.39 35 1.27 64 >30 6 0.28 36 0.28 65 6.68 7 0.44 37 1.59 66 >30 8 0.52 38 1.04 67 12.3 9 0.61 39 >100 68 3.78 10 0.64 40 >100 69 19.4 11 0.68 41 36.61 70 7.48 12 1.71 42 >100 71 23.4 13 1.72 43 49.69 72 38.5 14 1.92 44 22.13 73 1.07 15 1.85 45 1.19 74 0.20 16 2.49 46 2.87 75 0.035-0.045 17 3.85 47 4.43 76 8.0 18 3.07 48 4.99 77 2.31 19 4.28 49 6.56 78 10.12 20 9.29 50 1.2 81 0.0583 21 0.21 51 0.62 82 0.0353 22 17.50 52 1.62 83 19.1 23 18.5 53 0.18 84 0.275 24 19.1 54 0.24 85 0.832 25 23.7 55 11.08 86 20.85 27 0.13 56 0.7 87 >22.6 28 0.21 57 15.47 88 >100 29 0.31 58 0.33 89 >100 30 0.14 59 0.15

HPLC (high-performance liquid chromatography) method for measuring PSA inhibitory activity

We determined the IC ₅₀ value (concentration causing 50% inhibition) of compounds with potential PSA inhibitory activity after 2 h pretreatment with PSA. The substrate used was a 10-amino acid peptide, which was synthesized on an ABI 433 peptide synthesizer using the FMOC (fluorenyl methoxycarbonyl protecting group) method with HBTU (O-benzotriazol-1-yl-N,N,N',N'tetramethyluronium hexafluorophosphate) activation. The standard synthetic steps of the instrument were used. Cleavage of the peptide from the resin and removal of the protecting groups were performed with 80% trifluoroacetic acid (TFA), 5% thioanisole, 7.5% phenol, and 2.5% ethanedithiol. The crude peptide was purified by C-18 reversed-phase HPLC and characterized by amino acid analysis and electrospray mass spectrometry. The peptide substrate amino acid sequence was used to model the experimentally suggested interaction between PSA and bovine milk casein. The peptide amino acid sequence is SGAWYYVPLG [sequence 1] (serine-glycine-alanine-tryptophan-tyrosine-tyrosine-valine-proline-leucine-glycine), with a molecular weight of 1111.1 g/mol. The substrate was stored as a lyophilized solid at -20°C before regeneration. The substrate was regenerated in phosphate buffered saline (PBS) at a concentration of 0.1 mg/ml before assay.

The lyophilized substrate and PBS solution are removed from the freezer and allowed to warm to room temperature for at least 30 minutes before use.

Weigh out 1-5 mg of the compounds to be tested (in solid or oil form) and dilute them with DMSO to prepare a primary stock solution with a concentration of 300 pmol/l. For each compound tested for PSA inhibition, 1:10 dilution series are prepared from the primary stock solution with DMSO, thus obtaining stock solutions with the following concentrations: 30, 3, 0.3 pmol/l. The primary stock solution is designated “A”, the other stock solutions are designated “B”, “C”, and “D”, which correspond to concentrations of 30, 3, and 0.3 pmol/l, respectively. The final concentrations of the compounds to be tested in the reaction tubes are 30, 3, 0.3, and 0.03 pmol/l, respectively.

The following table shows the composition of the tested samples, with the volume of each component, e.g. in units. Samples 11 - I8 contain the tested compound. Comparative samples C1 - C3 contain only enzyme and substrate, sample S1 contains only substrate, and sample P1 contains only enzyme.

Add 80 µl of substrate to all tubes except P1 (final concentration 72 pmol/l). Dilute 100 µl of prostate-specific antigen stock solution [PSA - protein concentration 0.72 mg/ml (25.3 pmol/l) in 50% PBS/glycerol stored at -20°C; molecular weight 28.433 g/mol; specific activity of PSA was experimentally determined to be 155 mmol/hr/mg protein] with 620 µl of 50% PBS/glycerol. Initiate the reaction by adding 10 µl of diluted PSA solution (final concentration 357 nmol/l) to all HPLC tubes except S1. Gently swirl the tubes to ensure proper mixing, cap them, and incubate at 37°C for 2 hours.

To freeze the reaction, 100 µl of 0.2% TFA/water (trifluoroacetic acid/water) was added to each tube.

116

300 30 3 0.3 μΜ μΜ μΜ μΜ The B C D DMSO sub. PSA volume 11 10 0 0 0 0 80 10 100 I2 6.6 0 0 0 3.4 80 10 100 I3 0 10 0 0 0 80 10 100 I4 0 6.4 0 0 3.4 80 10 100 I5 0 0 10 0 0 80 10 100 I6 0 0 6.4 0 3.4 80 10 100 I7 0 0 0 10 0 80 10 100 I8 0 0 0 6.4 3.4 80 10 100 C1 0 0 0 0 10 80 10 100 C2 0 0 0 0 10 80 10 100 C3 0 0 0 0 10 80 10 100 S1 0 0 0 0 10 80 0 100 P1 0 0 0 0 10 0 10 100

The samples are analyzed by HPLC method. The analysis is performed with the following equipment and experimental conditions: -Bechman #507 type automatic sample changer; #116 type programmable solvent unit; #168 type diode array detector; System Gold software version 7.0; injection volume: 100 ml; Column_ Vydac Coompany, Catalog number #214TP5415 (4.6 mmol X 250 mmol) C-4 solid phase for protein and peptide separations; Particle size: 5 μην, Pore diameter: 300 Å; Detector parameters: System Gold diode array scanner (#168 type), detection at 210 and 280 nm. Mobile phase:

Solvent A = 0.1% aqueous TFA solution

Solvent B = 0.1% acetonitrile TFA solution

117

The gradient release program is as follows:

Time %B 0 minutes 10% 5 minutes 10% 35 minutes 30% 36 minutes 10% 40 minutes 10%

Dissolution (retention) times:

Peak 1 (YVPLG): 8.5 ± 0.3 minutes

Peak 2 (SGAWY): 13 ± 0.3 minutes

Peak 3 (SGAWYY): 19 ± 0.3 minutes

Peak 4 (unchanged substrate, SGAWYYVPLG): 30 ± 0.3 min

See Figure 1 for the chromatogram.

Data processing

Each compound was first evaluated at final concentrations of 30, 10, and 1 pmol/L. Compounds with estimated IC ₅₀ values < 10 pmol/L were re-evaluated using an 8-point concentration-response curve (30, 10, 3, 1, 0.3, 0.2, 0.03, 0.01 μίτιόΙ/Ι). System Gold software integrates the chromatographic peaks and calculates the peak areas used for statistical evaluation. The results of the comparative measurements were checked for consistency within a single determination. The % inhibitory effect was plotted as a function of inhibitor concentration and the curve was processed by non-linear regression. Compounds that did not inhibit 50% at the highest concentration (usually 30 pmol/l) were assigned an IC ₅₀ value (> at the highest concentration). For compounds that inhibited >50% at the highest inhibitor concentration, the IC ₅₀ value was determined using the JMP™ [SAS Institute, Cary, NC] nonlinear regression method. Compounds that did not inhibit the cleavage of the substrate by PSA were designated as inactive (NA, not active).

Results of the determination by HPLC method:

Example # IC50 µmol/l 2 0.91 7 1.12 21 <30 22 0.45 23 20.10 26 0.70 27 0.84

Some of the compounds of the present invention were tested for their inhibitory activity against other serine protease enzymes. The colorimetric method described above was used to test for PSA inhibitory activity. A description of the method for measuring thrombin and tissue plasminogen activator (t-PA) inhibition can be found in the following publication: “Thrombin Inhibitory Compounds Related to Efegatran Containing Arginine Aldehyde”, Seminars in Thrombosis and Hemostasis, Vol. 22, No. 2, 1996.

Example # PSA thrombin t-PA

1.2 NA NA

I 19

As discussed above, the compounds of the present invention effectively inhibit the proteolytic activity of PSA, and thus, the present invention also provides a method of inhibiting the proteolytic activity of PSA in a mammal, comprising administering to said mammal in need thereof an amount of a compound of the present invention that inhibits the proteolytic activity of PSA.

The compounds of the present invention effectively inhibit the proteolytic activity of prostate-specific antigen and are therefore effective in treating prostate cancer in mammals. Preferably, the mammalian species treated by administering the compounds of the present invention is a human.

The measurement of serum PSA concentration is now widely used to monitor patients with prostate cancer as well as to detect or estimate the likelihood of prostate cancer. Serum PSA levels in healthy men are less than 4 ng/ml. A man is suspected of having prostate cancer if his PSA concentration is between about 4 ng/l and about 10 ng/l, or if his PSA concentration is increasing over time. A man with a PSA concentration greater than 10 ng/ml is almost certain to have prostate cancer. Accordingly, the compounds of the present invention can be used to prevent the development of prostate cancer, to delay the development of prostate cancer, or to reduce the likelihood of developing prostate cancer in men with elevated PSA levels, such as those with a family history of the disease.

The compounds of the present invention, when used alone, may also be an effective treatment for prostate cancer. In another embodiment of the present invention, the PSA inhibitors of the present invention are combined with other known treatment methods to provide effective treatment for prostate cancer.

Although the compound used in the methods of the present invention can be administered in its pure form, the compounds are typically administered in the form of pharmaceutical compositions comprising a pharmaceutically acceptable excipient and at least one active ingredient (the compound of the present invention). These pharmaceutical compositions contain from about 0.1% by weight to about 90.0% by weight of the compound of the present invention. These pharmaceutical compositions can be administered in a variety of ways, such as orally (by mouth), rectally (by rectum), topically, transdermally (by skin), buccally (by mouth), subcutaneously (by skin), intravenously (by vein), intramuscularly (by muscle), or intranasally (by nose). Many of the compounds used in the methods of the present invention are effective in either injectable, oral, or topical pharmaceutical compositions. Such pharmaceutical compositions may be prepared by methods well known to the pharmacist and contain at least one active compound. See, for example, Remington's Pharmaceutical Sciences, (16th Edition, 1980).

In preparing the pharmaceutical composition used in the method of the present invention, the active ingredient is usually mixed with or diluted with a filler or enclosed in a carrier, which may be a capsule, wafer, paper, or other shaped container. If the filler is also a diluent, it may be a solid, quasi-solid, or liquid material which serves as a carrier, vehicle, or medium for the active ingredient. Thus, the pharmaceutical compositions may be in the form of tablets, pills, powders, lozenges, wafers, cachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols (in solid form or in a liquid medium), ointments containing, for example, up to 10% by weight of the active ingredient, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

In the preparation of a pharmaceutical composition, it may be necessary to grind the active compound to a suitable particle size before mixing with other ingredients. If the active compound is substantially insoluble, it is usually ground to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size is usually adjusted by grinding so that the distribution of the compound throughout the pharmaceutical composition is substantially uniform.

Suitable carriers, fillers and diluents include lactose (milk sugar), dextrose, sucrose (cane sugar), sorbitol, mannitol, starches, acacia, calcium phosphate, alginates, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, alcohol, water, sugar syrup, and methylcellulose. The various dosage forms may also contain anti-caking agents such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl and propyl hydroxybenzoates; sweeteners; and flavoring agents. The pharmaceutical compositions of the present invention may be formulated by methods known per se in such a way that the absorption of the active ingredient after administration to the patient is rapid, slow, or delayed.

The compounds of the present invention can be administered transdermally using known transdermal delivery systems and excipients. Most preferably, the compound of the present invention is mixed with additives that enhance skin penetration, such as (but not limited to) propylene glycol, polyethylene glycol monolaurate, and aza-alkane-2-one derivatives, and the mixture is then applied to a patch or similar delivery system. The transdermal dosage form may also contain additional excipients, such as gelling agents, emulsifiers, and buffering agents, as desired.

For topical application, the compound of the present invention can ideally be mixed with a wide variety of excipients to form a high viscosity liquid or cream-like preparation.

For oral administration, the compound of the present invention can ideally be mixed with carriers and diluents and then compressed into tablets or filled into gelatin capsules.

The pharmaceutical compositions are preferably presented in unit dosage form, each dosage containing from about 0.05 to about 500 mg, usually from about 1.0 to about 100 mg, more often from about 1.0 to about 50 mg, of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable for unitary administration to a human or other mammal, each dosage unit containing a predetermined quantity of the active ingredient which, when mixed with a suitable pharmaceutical excipient, produces the desired therapeutic effect.

123

The active compounds are generally effective over a wide range of doses. Typical daily doses are, for example, about 0.01 to about 500 mg/kg, preferably about 0.01 to about 30 mg/kg, more preferably about 0.01 to about 10 mg/kg, based on body weight. In the treatment of adults, a preferred daily dose range is about 0.1 to about 5 mg/kg, administered in single or divided doses. It will be appreciated, however, that the amount of compound actually administered will be determined by the physician in the light of the relevant circumstances, including the nature of the condition being treated, the dosage form chosen, the compound or compounds actually administered, the age and weight of the individual being treated, the response to treatment, and the severity of the symptoms, and therefore the above dosage ranges are not intended to limit the scope of the invention in any way. There may be cases in which doses lower than the lower end of the aforementioned dosage range are more than adequate, while in other cases doses higher than those indicated may be used without adverse side effects if these larger doses are divided into several smaller doses administered throughout the day.

In order to more fully illustrate the implementation of the present invention, the following are examples of dosage forms. The examples are for illustrative purposes only and are not intended to limit the scope of the patent in any way.

The compositions may contain any of the compounds of the present invention as the active ingredient (compound).

124

1. Pharmaceutical preparation

We prepare hard gelatin capsules containing the following ingredients:

Active ingredient: starch kernel stearate

Quantity (mg/capsule)

10.0

325.0

5.0

The above ingredients are mixed and filled into hard gelatin capsules of 340 oregano.

2. Pharmaceutical preparation

We prepare tablets with the following composition:

Component Quantity (mg/tablet) active ingredient 100.0 microcrystalline cellulose 125.0 colloidal silica 10.0 stearic acid 5.0 The above ingredients mixed and weighed 240 mg

pressed into tablets.

3. Pharmaceutical preparation

We prepare a dry powder dosage form containing the following ingredients:

Ingredient weight% active ingredient 5 lactose 95

The active ingredient is mixed with lactose and the mixture is filled into a dry powder inhaler.

125

4, Pharmaceutical preparation

Tablets containing 30 mg of active ingredient each are prepared as follows:

Quantity

Ingredients (mg/capsule) active ingredient 30.0 starch 45.0 microcrystalline cellulose 125.0 polyvinylpyrrolidone (in 10% aqueous solution) 4.0 sodium carboxymethyl starch 4.5 magnesium stearate 0.5 talc 1.0 total 120

The active ingredient, starch and cellulose are screened through a 20 mesh US sieve and mixed thoroughly. The polyvinylpyrrolidone solution is mixed with the resulting powder, which is then passed through a 16 mesh US sieve. The resulting granules are dried at 50-60°C and passed through a 16 mesh US sieve. Sodium carboxymethyl starch, magnesium stearate and talc, previously screened through a 30 mesh US sieve, are added to the granules and the mixture is mixed and formed into tablets weighing 120 mg each on a tabletting machine.

5. Pharmaceutical preparation

Capsules containing 40 mg of active ingredient each are prepared as follows:

126

Ingredient active ingredient starch magnesium stearate total

Quantity (mg/capsule)

40.0

109.0

JLO

150.0

The active ingredient, cellulose, starch, and magnesium stearate are mixed, passed through a 20-mesh U.S. sieve, and filled into hard gelatin capsules in 150 mg portions.

6. Pharmaceutical preparation Ingredient Quantity active ingredient 25 mg supplemented with saturated fatty acid glycerides to 2000 mg The active ingredient is screened through a 60 mesh US sieve and suspended in a mixture of saturated fatty acid glycerides previously melted with as little heat as possible. The mixture is then poured into a cone mold of nominal capacity 2.0 g and allowed to cool.

7. Pharmaceutical preparation

Preparation of 5.0 ml suspensions containing 50 mg of active substance per dose:

Component Quantity active ingredient 50.0 mg xanthan gum 4.0 mg sodium carboxymethyl cellulose (11%) microcrystalline cellulose (89%) 50.0 mg sucrose 1.75 grams

127

sodium benzoate 10.0 mg fragrance and colorant optionally supplemented with purified water to 5 ml

The active ingredient, sucrose and xanthan gum are combined, passed through a 10-mesh US sieve, and mixed with the previously prepared aqueous solution of microcrystalline cellulose and sodium carboxymethyl cellulose. Then, while stirring, the sodium benzoate, fragrance and colorant diluted with a small amount of water are added. The mixture is then made up to volume with water.

8. Pharmaceutical preparation

Preparation of capsules containing 15 mg of active ingredient each:

Ingredient active ingredient starch magnesium stearate total

Quantity (mg/capsule)

40.0

109.0

1.0

150.0

The active ingredient, cellulose, starch, and magnesium stearate are blended, passed through a 20-mesh U.S. sieve, and filled into 425 mg hard gelatin capsules.

9. Pharmaceutical preparation

An intravenous dosage form can be prepared as follows:

128

Ingredient active ingredient isotonic saline solution

Quantity 250.0 g 1000 ml

10. Pharmaceutical preparation

A topical dosage form can be prepared as follows:

Ingredient active ingredient emulsifying wax liquid paraffin

Quantity 1 - 10 g 30 g supplemented with 20 g white soft paraffin to 100 g

The white soft paraffin is heated until melted. Then the liquid paraffin and the emulsifying wax are added and mixed until completely dissolved. The active ingredient is added and mixing is continued until it is completely distributed. The mixture is then cooled and solidified.

11, Pharmaceutical preparation

Sublingual (placed under the tongue) or buccal (dissolved in the mouth) tablets containing 10 mg of active ingredient each can be prepared as follows:

Component Quantity per tablet active ingredient 10.0 mg glycerin 210.5 mg water 143.0 mg sodium citrate 4.5 mg polyvinyl alcohol 26.5 mg po I i vin il-p írűl id ο n 15.5 mg

all

410.0 mg

129

The glyceryl, water, sodium citrate, polyvinyl alcohol, and polyvinyl pyrrolidone are mixed with constant stirring, while maintaining the temperature of the mixture at about 90°C. When the polymers have dissolved, the solution is cooled to a temperature of about 50-55°C and the active ingredient is slowly stirred in. The homogeneous mixture is poured into molds made of neutral material, thus obtaining a diffusion matrix (carrier) containing the active ingredient with a thickness of about 2-4 mm, from which tablets of appropriate size are then cut.

Another preferred dosage form for use in the methods of the present invention is a transdermal delivery device ("patch"). These transdermal patches can be used to provide controlled, continuous or intermittent delivery of the compounds of the present invention. Methods of making and using transdermal patches for the delivery of therapeutic agents are known in the art. See, for example, U.S. Patent Application Serial No. 5,023,252, published June 11, 1991, which is incorporated herein by reference. Such patches can be prepared for continuous, bolus or as-needed delivery of the therapeutic agents.

In the generally preferred indirect methods, the dosage form is usually formulated by converting the water-soluble active ingredient into a fat-soluble active ingredient, a so-called "prodrug" (an inactive derivative that is converted into the active ingredient when it enters the body), thereby allowing the absorption of the active ingredient to be delayed. The delay in absorption is usually achieved by providing the hydroxyl, carbonyl, sulfate, and primary amine groups in the active ingredient with protecting groups, making the compound more fat-soluble, which thus penetrates the blood-brain barrier more easily. According to another method, the delivery of water-soluble active ingredients into the body can be enhanced by intra-arterial infusion of hypertonic (higher salt concentration than blood) solutions, which temporarily opens the blood-brain barrier.

The type of dosage form used to administer the compounds used in the methods of the present invention may be determined by the structure of the compound employed, the pharmacokinetic behavior desired from the dosage form and compound(s), and the condition of the patient.

Claims (42)

Patent claims 1. A compound of formula I, and pharmaceutically acceptable salts and solvates thereof, wherein the R ¹ is an aryl, aryl-(C 1-6 alkylene) group, wherein the aryl group or the ring of the aryl-(C 1-6 alkylene) group may be substituted with one or more groups independently selected from the following: halide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene groups; phthalimido, or one of the groups of formulae (a) to (e), wherein the R ⁴ is a hydrogen atom, a C 1-6 alkyl, aryl, or heterocycle; wherein the aryl or heterocycle may be optionally substituted with one or two groups selected from the following: halide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene; R ⁵ is hydrogen, C 1-4 alkyl, C 3-7 cycloalkyl, C 1-4 alkyloxycarbonyl, phenyl, or naphthyl, wherein the phenyl or naphthyl group may be optionally substituted with one or two groups selected from the following: C 1-4 alkyl, C 1-4 alkyloxy, halide, hydroxyl, cyano, carbamoyl, amino, mono(C 1-4 alkyl)amino, di(C 14 alkylamino, C 1-4 alkylsulfonylamino, and nitro; R ⁶ is a hydrogen atom, a C 1-4 alkyl group optionally substituted with one or two groups selected from the following: hydroxyl, protected carboxyl, carbamoyl, benzylthio or C 1-4 alkylthio group; or a phenyl group optionally substituted with one or two groups selected independently from the following: C 1-4 alkyl group, C 1-4 alkyloxy group, halide, hydroxyl, cyano, carbamoyl, amino, mono(C 1-4 alkyl)amino, di(C 1-4 alkyl)amino and nitro group, or C 1-4 alkyloxycarbonyl group; R ⁷ is a C 1-4 alkyloxycarbonyl, benzyloxycarbonyl, benzoyl group, wherein the phenyl group of the benzyloxycarbonyl or benzoyl group may be optionally substituted with one or two groups independently selected from the following: C 1-4 alkyl, C 1-4 alkyloxy, halide, cyano, nitro, amino, carbamoyl, hydroxyl, mono(C 1-4 alkyl)amino and di(C 1-4 alkyl)amino groups; the groups of general formula R ⁸ and R ⁹ independently represent a hydrogen atom, a C 1-5 alkanoyloxy, a benzoyloxy group, which may be substituted by one or two groups independently selected from the following: a C 1-4 alkyl, a C 1-4 alkyloxy, a halide, a cyano, a nitro, an amino, or a C 1-4 alkyloxycarbonyl group; a benzyloxy, a diphenylmethoxy, or a triphenylmethoxy group; with the proviso that only one of the groups of general formula R ⁸ and R ⁹ may represent a hydrogen atom; R ² is a C 2-6 alkenyl, aryl, or heterocyclic group, wherein the aryl or heterocyclic group may be optionally substituted with one or two groups independently selected from the group consisting of halide, N-oxide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxyethylene, and hydroxymethylene; -CH ₂ CH ₂ R ¹⁰ is a group of general formula, in which the R ¹⁰ is carboxyl, aryl or heterocycle, wherein the aryl or heterocycle may be optionally substituted with one or two groups independently selected from the group consisting of halide, n-oxide, C1-6 alkyloxy, methoxycarbonyl, phenyl, C1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene and hydroxymethylene; -CH = C(R ¹² )-R ¹¹ group, in which the R ¹¹ is hydrogen or phenyl, and R ¹² is nitrile, aryl or heterocycle, wherein the aryl or heterocycle may be optionally substituted with one or two groups independently selected from halide, n-oxide, C1-6 alkyloxy, methoxycarbonyl, phenyl, C1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene134 group; with the proviso that when R ¹¹ is phenyl, R ¹² is aryl; or -CsC-R ¹³ , wherein R ¹³ is an aryl group or a heterocycle, wherein the aryl group or the heterocycle may be optionally substituted with one or two groups independently selected from the group consisting of halide, n-oxide, C1-6 alkyloxy, methoxycarbonyl, phenyl, C1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene; and R ³ is a heterocycle, CO ₂ R ¹⁴ , or (f) a group, wherein R ¹⁴ is a C 2-6 alkenyl, C 1-6 alkyl, C 1-6 alkyl substituted with halogen, C 3-7 cycloalkyl, substituted C 3-7 cycloalkyl, aryl-substituted C 1-6 alkyl or heterocycle, wherein the aryl group or the heterocycle or the ring of the aryl-substituted C 1-6 alkylene group may be optionally substituted with one or two groups independently selected from the following: halide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, and hydroxymethylene; R ¹⁵ is an aryl group or a heterocyclic substituted C 1-6 alkylene group, wherein the ring of the aryl group or the heterocyclic substituted C 1-6 alkylene group may be optionally substituted with one or two groups independently selected from the group consisting of halide, C 1-6 alkyloxy, methoxycarbonyl, phenyl, C 1-6 alkyl, carboxyl, nitro, acetyl, formyl, carboxymethylene, trifluoromethyl, and hydroxymethylene; 2. A compound according to claim 1, and pharmaceutically acceptable salts and solvates thereof, wherein the R ¹ is a benzyl, phthalimido, or (g) group, wherein R ⁵ represents a hydrogen atom, a C 1-4 alkyl group, or a phenyl group; R ⁶ represents a hydrogen atom, an isopropyl group, or a phenyl group; The group of general formula R ² means a phenyl-C 2-C 4 alkenyl group, -CH ₂ CH ₂ R ¹⁰ , -CH = C(R ¹¹ )-R ¹² , or a group of general formula C^CR ¹³ , in which the R ¹⁰ is a carboxyl or phenyl group; R ¹¹ represents a hydrogen atom or a phenyl group; R ¹² is cyano, phenyl, naphthyl, 2-furanyl, or 3-furanyl, pyridinyl, pyrimidinyl, or quinolinyl; and wherein the phenyl group may be optionally substituted with a C 1-4 alkyl, C 1-4 alkyloxy, or nitro group, and wherein the pyridinyl group may have an N-oxide; R ¹³ is a phenyl group; R ^{3 *} is a heterocyclic group, CO ₂ R ¹⁴ , or (f) a group, wherein the heterocyclic group is pyridinyl, pyrimidinyl, 1,3,5-triazinyl, quinazolinyl or benzoxazolyl, and wherein said heterocyclic group is optionally substituted independently with one or two nitro, trifluoromethyl, C 1-4 alkyloxy or phenyl groups; R ¹⁴ is a C 2-4 alkenyl, C 1-4 alkyl, C 1-4 halogenated alkyl, C 4-7 cycloalkyl, 2-isopropyl-5-methylcyclohexanyl, benzyl or phenyl group; wherein the phenyl group may be optionally substituted with a halide, C 1-4 alkyloxy, carbomethoxy or nitro group; R ¹⁵ is a phenyl, naphthyl, 2-furanylmethyl, or 3-furanylmethyl group, wherein the phenyl group may be optionally substituted with one to four groups, which may independently be halide or trifluoromethyl groups. 3. A compound of formula (H) according to claim 2, and pharmaceutically acceptable salts and solvates thereof, wherein R ³ represents a heterocyclic group, CO ₂ R ¹⁴ or (f) a group in which the heterocyclic group is pyridinyl, pyrimidinyl, 1,3,5-triazinyl, quinazolinyl, or benzoxazolyl, and in which the 137 said heterocyclic group is optionally substituted independently with one or two nitro, trifluoromethyl, C1-C4 alkyloxy, or phenyl groups; R ¹⁴ is a C 2-4 alkenyl, C 1-4 alkyl, C 1-4 halogenated alkyl, C 4-7 cycloalkyl, 2-isopropyl-5-methylcyclohexanyl, benzyl or phenyl group; R ¹⁴ is a C 2-4 alkenyl, C 1-4 alkyl, C 1-4 halogenated alkyl, C 4-7 cycloalkyl, 2-isopropyl-5-methylcyclohexanyl, benzyl or phenyl group; wherein the phenyl group may be optionally substituted with a halide, C 1-4 alkyloxy, carbomethoxy or nitro group; R ¹⁵ is phenyl, naphthyl, 2-furanylmethyl, or 3-furanylmethyl, wherein the phenyl group may be optionally substituted independently with one to four halide or trifluoromethyl groups. 4-methoxycarbonylphenoxycarbonyl, 4-methoxyphenoxycarbonyl or 1-propylenoxycarbonyl; which process consists in reacting a 3-(4S-phenyloxazolidin-2-on-3-yl)-4-ethenylazetidinone derivative of general formula IX, in which the The group of general formula R ²⁰ represents a tributylstannyl or B(OH) ₂ - group, in the presence of a catalyst, a functional group is introduced using the electrophilic reagent of general formula XR ¹⁶ , in which the group of general formula X represents a leaving group, and the substituted azetidinone derivative of general formula Ia is prepared by acylation of the thus formed compound of general formula XI. A therapeutically effective amount of a compound according to claim 4 is administered. 4. A compound of formula Ib, and pharmaceutically acceptable salts and solvates thereof, wherein The group of general formula R represents a naphthyl, benzyl, phenyl or furfuryl group; in which the aromatic ring of the naphthyl, phenyl or benzyl group may be substituted with one to four halide, trifluoromethyl or C1-C6 alkyl groups. ^{5 * *} 5. A method for inhibiting the proteolytic activity of prostate-specific antigen, which method comprises administering to a mammal in need of said inhibitory effect a pharmaceutically effective amount of a compound according to claim 1. 6. A method for inhibiting the proteolytic activity of prostate-specific antigen, which method comprises administering to a mammal in need of said inhibitory effect a pharmaceutically effective amount of a compound according to claim 2. 7. A method for inhibiting the proteolytic activity of prostate-specific antigen, which method comprises administering to a mammal in need of said inhibitory effect a pharmaceutically effective amount of a compound according to claim 3. 8. A method for inhibiting the proteolytic activity of prostate-specific antigen, which method comprises administering to a mammal in need of said inhibitory effect a pharmaceutically effective amount of a compound according to claim 4. 9. A method for treating prostate cancer, comprising administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 1. 10. A method for treating prostate cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 2. 11. A method for treating prostate cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 3. 12. A method for treating prostate cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 4. 13. A method for treating prostate cancer, comprising administering to a mammal in need of said treatment a compound that inhibits prostate-specific antigen. 14. A method for treating benign prostatic hyperplasia, said method comprising administering to a mammal in need of said treatment a compound that inhibits prostate-specific antigen. 15. A method for treating prostate cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 1. 16. A method for treating prostate cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 2. 140 ........... 17. A method for treating prostate cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 3. 18. A method for treating prostate cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 4. 19. A method for treating breast cancer, comprising administering to a mammal in need of said treatment a compound that inhibits prostate-specific antigen. 20. A method for treating breast cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 1. 21. A method for treating breast cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 2. 22. A method for treating breast cancer, which method comprises administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 3. 23. A method for treating breast cancer, the method comprising administering to a mammal in need of said treatment: 24. A method for treating prostate cancer metastases, comprising administering to a mammal in need of said treatment a compound that inhibits prostate-specific antigen. 25. A method for treating prostate cancer metastases, comprising administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 1. 26. A method for treating prostate cancer metastases, comprising administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 2. 27. A method for treating prostate cancer metastases, comprising administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 3. 28. A method for treating prostate cancer metastases, comprising administering to a mammal in need of said treatment a therapeutically effective amount of a compound according to claim 4. 29. A pharmaceutical composition comprising a compound of claim 1 in combination with one or more pharmaceutically acceptable carriers, diluents or excipients. (42 *··’ ***' ·«♦* *··* ·*· 30. A pharmaceutical composition comprising a compound according to claim 2 in combination with one or more pharmaceutically acceptable carriers, diluents or excipients. 31. A pharmaceutical composition comprising a compound according to claim 3 in combination with one or more pharmaceutically acceptable carriers, diluents or excipients. 32. A pharmaceutical composition comprising a compound according to claim 4 in combination with one or more pharmaceutically acceptable carriers, diluents or excipients. 33. A process for the preparation of a compound of formula Ia and its pharmaceutically acceptable salts and solvates, wherein the R ¹⁶ is a group of general formula: phenyl, 2-nitrophenyl, 4-nitrophenyl, naphthyl, 3-nitro-1-naphthyl, 4-nitro-1-naphthyl, 2-quinolinyl, 4-quinolinyl, 7-quinolinyl, 1-isoquinolinyl, 3-isoquinolinyl, 8-isoquinolinyl, 2-benzothiazolyl, 2-benzoxazolyl, 2-thienyl, 3-furanyl, 3-pyridinyl, N-oxy-3-pyridinyl, 5-pyrimidinyl, or 3,5-dimethylphenyl; and R ¹⁷ is 1-phenoxycarbonyl, 4-chlorophenoxycarbonyl, 4-fluorophenoxycarbonyl, benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-chlorobutyloxycarbonyl, 4-methoxycarbonylphenoxycarbonyl, 4-methoxyphenoxycarbonyl or 1-propylenoxycarbonyl; which process consists of reacting 3-(4S-phenyloxazolidin-2-on-3-yl)-4-ethenyl-azetidinone derivative of general formula IX, in which the R ²⁰ is a tributylstannyl or B(OH) ₂ - group, a functional group is introduced in the presence of a catalyst using the electrophilic reagent XR ¹⁶ , in which the group X is a leaving group, and the thus formed compound XI is acylated to produce the substituted azetidinone derivative Ia. 34. Compound of Formula IXa. 35. Compound of Formula IXc. 36. A compound of formula VI, wherein R ¹⁸ is a trialkylsilanyl group and R ¹⁹ is a nitrogen protecting group. 37. A compound of formula VII, wherein R ¹⁹ is a nitrogen protecting group. 38. Compound of Formula VIII. 39. A process for the preparation of a compound of formula Ia, and pharmaceutically acceptable salts and solvates thereof, wherein the R ¹⁶ is a group of general formula: phenyl, 2-nitrophenyl, 4-nitrophenyl, naphthyl, 3-nitro-1-naphthyl, 4-nitro-1-naphthyl, 2-quinolinyl, 4-quinolinyl, 7-quinolinyl, 1-isoquinolinyl, 3-isoquinolinyl, 8-isoquinolinyl, 2-benzothiazolyl, 2-benzoxazolyl, 2-thienyl, 3-furanyl, 3-pyridinyl, N-oxy-3-pyridiηyl, 5-pyrimidinyl, or 3,5-dimethylphenyl; and R ¹⁷ is 1-phenoxycarbonyl, 4-chlorophenoxycarbonyl, 4-fluorophenoxycarbonyl, benzyloxycarbonyl, 4-nitrobenzyloxycarbonyl, 4-chlorobutyloxycarbonyl, Γ IK / 40. Compound of Formula IX. 41. A compound of formula VIII, wherein R ¹⁹ is a nitrogen protecting group. 42. (vi) A compound of formula.