EA037916B1 - Beta-lactamase inhibitor compounds - Google Patents

Abstract

The invention relates to compounds which are beta-lactamase inhibitors. The compounds and their pharmaceutically acceptable salts in combination with beta-lactam antibiotics are useful for the treatment of bacterial infections, including infections caused by drug resistant organisms, including multi-drug resistant organisms. The invention includes compounds of the following Formulae:or a pharmaceutically acceptable salt thereof, wherein the values of R1, R2, R3, R4, R5 and R6 are described herein.

Description

Related applications

This application claims priority based on U.S. Provisional Application No.

62 / 395,464, filed September 16, 2016, and US Provisional Application No. 62 / 456,423, filed February 8, 2017, the contents of each of which are incorporated herein by reference.

Technology area

The invention relates to novel oral beta-lactamase inhibitors, pharmaceutical compositions containing them and methods of using them. In addition, the present invention relates to therapeutic methods for treating bacterial infections, including overcoming bacterial antibiotic resistance.

State of the art

The international microbiological and infectious disease community continues to express grave concern that the continued development of antibacterial resistance could lead to bacterial strains against which currently available antibacterial drugs will be ineffective, leading to significant morbidity and mortality. In general, bacterial pathogens can be classified as gram-positive or gram-negative pathogens. Antibiotics effective against both gram-positive and gram-negative pathogens are generally considered to have a broad spectrum of activity.

Beta-lactam antibiotics are needed to fight bacterial infection. Beta-lactams are a wide class of drugs, all of which contain beta-lactams in their basic molecular structure and generally show efficacy against a wide range of gram-positive and gram-negative bacteria by inhibiting bacterial cell wall synthesis. Since the drug target has no eukaryotic counterparts, their toxicity is low and they are generally well tolerated. They remain some of the most widely prescribed, safe and effective medicines available to fight bacterial infection. However, their effectiveness is limited by highly resistant infectious strains such as methicillin-resistant Staphylococcus aureus (MRSA) strains and multidrug-resistant (MDR) strains of Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli, Klebsiella pneumoniae, and others. Such resistant bacteria are the main causes of morbidity and mortality in patients. Helfand, β-lactams Against Emerging 'Superbugs': Progress and Pitfalls, Expert Rev. Clin. Pharmacol. 1 (4): 559-571 (2008).

Beta-lactam antibiotics, alone and in combination with beta-lactamase inhibitors, continue to represent an important part of the antibacterial agents used to combat disease. Resistance to beta-lactam in gram-negative infections is mainly due to beta-lactamase activity, and significant dependence on beta-lactam antibiotics has led to a diversification and increased prevalence of beta-lactamases. These beta-lactamases cause resistance to even the newest beta-lactam antibiotics. Llarrull, et al., The Future of Beta-Lactams, Current Opinion in Microbiology, 13: 551-557 (2010).

The main threat to the effectiveness of these drugs is the growing prevalence of extended spectrum beta-lactamases (ESBLs). Beta-lactamases are enzymes that are produced by some bacteria and open the ring of the beta-lactam part of the beta-lactam antibiotic, thereby deactivating it. There are currently four classes of beta-lactamases, designated as class A, class B, class C and class D. Class A, class C and class D beta-lactamases are serine beta-lactamases, and class B beta-lactamases are metallo-beta-lactamase (MBL). Bush & Jacoby, Updated Functional Classification of β-Lactamases, Antimicrobial Agents and Chemotherapy, 54 (3): 969-976 (March 2010).

Several beta-lactamase inhibitors have been developed to increase the effectiveness of beta-lactam antibiotics. However, currently available beta-lactamase inhibitors are in many cases insufficient to counteract the ever-growing variety of beta-lactamases. The three most commonly used agents against serine beta-lactamases - clavulanic acid, tazobactam and sulbactam - are active only against certain class A enzymes, which seriously limits their practical value. In addition, new beta-lactamase inhibitors recently approved or currently in clinical trials, such as avibactam and MK-7655, are available for intravenous use only and act primarily on class A and C enzymes, with minimal efficacy against beta-lactamase. Class D. Lactamase Bebrone, et al., Current Challenges in Antimicrobial Chemotherapy: Focus on β-Lactamase Inhibition, Drugs, 70 (6): 651-679 (2010). While these agents represent a significant improvement over the currently available beta-lactamase inhibitors, agents effective against all three classes of beta-lactamase serine are desirable to combat the significant beta-lactam resistance currently observed, with the added benefit of available an orally effective dosage form for use outside the hospital. Does not currently exist

- 1,037916 approved oral beta-lactamase inhibitors that are effective against class C or class D beta-lactamases, while levels of resistance to common antibiotics continue to rise.

Compounds similar to those described herein, also having broad beta-lactamase inhibition profiles (effective against most class A, class C and class D beta-lactamases), have been described in WO 2013/150296. This patent application relates to compounds of the following formula:

a

While the compounds described in WO 2013/150296 represent a significant improvement in the range of beta-lactamase inhibitors currently available on the market or in clinical practice, the compounds described in this application can only be administered intravenously (i.v. c) because they are not orally bioavailable. Moreover, the compounds disclosed therein cannot be made orally bioavailable by using a prodrug at a sulfate activating group in the molecule. Consequently, these effective beta-lactamase inhibitors are limited to intravenous or parenteral administration, which usually only occurs in a hospital setting. Consequently, for patients with severe, persistent infections who are otherwise healthy and do not require hospitalization, or patients who could be discharged from the hospital but who would benefit from outpatient antibiotic treatment (also known as stepped therapy with switching to the oral route), there are only limited treatment options. The compounds described in WO 2013/150296 can provide patients with more effective and wider inhibition of beta-lactamases than any of the currently known drugs, but these compounds currently require intravenous administration in a hospital setting.

There is an urgent need for a new broad-spectrum oral beta-lactamase inhibitor that would provide significant benefit to patients infected with resistant pathogens who could be treated outside the hospital or hospitalized patients who may not have reliable venous access. These patients may have severe, complicated infections with pathogens producing one or more beta-lactamases but may not require hospital treatment or recover from infections that were initially successfully treated with a IV generation beta-lactam / beta-lactamase inhibitor combination but would benefit from further out-of-hospital beta-lactam / beta-lactamase inhibitor combination therapy, which would only be possible with an orally active broad-spectrum beta-lactamase inhibitor such as described herein.

In addition, for patients with persistent bacterial infections who require hospitalization for initial treatment, the compounds of the present invention, as described in formulas (Ia), (IIa), (IIIa), (IVa) and (Va), can be administered intravenously in hospital conditions until the patient is stable enough to continue treatment on an outpatient basis. After discharge, the patient can continue therapy with the same drug by administering a compound according to any of formulas (I), (II), (III), (IV) or (V) on an outpatient basis, while continuing continuous treatment until the bacterial infection will not be cured. Currently, there are no class A, C and D β-lactamase inhibitors that could be initially administered IV with the added benefit of oral administration when the patient feels well enough to be discharged from the hospital. The ability for the physician to tailor treatment to the patient's needs would allow earlier discharges of patients requiring hospitalization and significantly reduce treatment costs in general, avoiding long hospital stays.

There is an urgent need for new orally active beta-lactamase inhibitors effective against more than one class A, C and D beta-lactamase.

Brief description of the invention

The invention relates to compounds that are orally available inhibitors of beta-lactamases. The compounds and their pharmaceutically acceptable salts, in combination with beta-lactam antibiotics, are suitable for the treatment of bacterial infections, including those caused by drug-resistant organisms, including multidrug-resistant organisms.

More specifically, the invention relates to compounds of the following formulas:

- 2 037916

or their pharmaceutically acceptable salts, where R ^{1 is} selected from —CN, —CH2NH2, —C (O) NH2, where R ¹¹ is hydrogen or —C (O) NH _2; or

R ^{1 is} selected from —C (O) NR ⁷ R ⁸ , where R ⁷ is hydrogen and R ⁸ is:

1) phenyl optionally substituted with C ₁ -C ₃ -α-alkyl or C ₁ -C ₃ -alkyl-NH ₂ ,

2) C ₁ -C ₃ -alkyl or

3) C ₁ -C ₃ alkoxy, where each alkyl or alkoxy of R ^{8 is} optionally and independently substituted with C 3 -C 6 cycloalkyl, —CN, —OH, —NH ₂ , —SO ₂ NH ₂ , —NHSO ₂ NH ₂ , - C (O) NH ₂ , -NHC (O) (C ₁ -C ₃ -αalkyl), pyrazinyl, oxytanyl, oxazolyl or pyrrolidinyl optionally substituted with one or more carboxyl, fluorine or -C (O) O (C ₁ -C ₆ -alkyl);

R ² is C ₁ -C ₃ -alkyl;

R ³ is C ₁ -C ₃ -alkyl;

R4 and R ⁵ independently represent hydrogen, methyl or fluoro; and

R ⁶ is hydrogen, C ₁ -C ₁₂ -alkyl, C ₁ -C ₄ -alkyl-OC (O) - (NHC ₁ -C ₆ -alkyl) -C (O) C ₁ -C ₃ alkoxy, C ₁ -C ₄ -alkyl-OC (O) -C ₁ -C ₄ -alkyl or C ₁ -C ₄ -alkyl-OC (O) -C ₁ -C ₃ -alkoxy.

The present invention also relates to the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in combination with a beta-lactam antibiotic, in the treatment of a bacterial infection in a subject in need thereof.

Detailed description of the invention

In one aspect, the invention relates to a compound of formula (I) which is an oral beta-lactamase inhibitor:

or a pharmaceutically acceptable salt thereof, where

R ¹ is -C (O) NR ⁷ R ⁸ , -CN, phenyl, 5-7 membered heteroaryl, -C (O) NR'NR'C (O) R ⁹ , -C (O) NR'OR ¹⁰ or a C1-C6 alkyl group, where the alkyl group is substituted by 1-3 groups consisting of halogen, C1-C3 alkoxy, -OH, -CN, -NR ⁷ R ⁸ , -NR7COR ⁹ , 5-7 membered heteroaryl and 5-7 membered heterocyclyl, and where phenyl and heteroaryl, represented among R ¹ , are optionally and independently substituted by 1-3 groups selected from halogen, -OH, C1-C ₃ -αalkoxy, -CN, -NR7R ⁸ and - CONR7R ⁸ ;

R ² and R ^{3 are} independently selected from hydrogen, halogen, C ₁ -C ₃ -alkyl and C ₃ -C ₆ -cycloalkyl;

R ⁴ and R ^{5 are} independently selected from hydrogen, halogen, —CN, —CO2R ⁹ , C ₁ -C ₃ alkyl, and C ₁ -C ₃ haloalkyl;

R ⁶ is hydrogen, C1-C12-alkyl, C1-C4-alkyl-C1-C3-α-alkoxy- (NR'C1-C6-alkyl) -C1-C3 alkoxy, C1-C4-alkyl-C1-C3-alkoxy- C1-C3 alkoxy, C2-C12 alkenyl, C3-C10 cycloalkyl, 5-7 membered heteroaryl and 5-7 membered heterocyclyl, where alkyl, alkenyl, cycloalkyl, heteroaryl and heterocyclyl are optionally and independently substituted with 1-6 groups selected from carboxyl, halogen, C1-C6 alkoxy, C1-C6 alkyl and phenyl. Alternatively, R ⁶ represents a C1-C12 alkyl, C ₁ -C ₄ -alkyl-C ₁ -C ₃ - alkoxy-3 037 916 (NR'S ₁ -C ₆ -alkyl) -C ₁ -C ₃ - alkoxy, C ₁ -C ₄ -alkyl-C ₁ -C ₃ -alkoxy-C ₁ -C ₃ -alkoxy, C ₂ -C _12- alkenyl, C ₃ C ₁₀ -cycloalkyl, 5-7 membered heteroaryl and 5- 7-membered heterocyclyl wherein alkyl, alkenyl, cycloalkyl, heteroaryl, and heterocyclyl are optionally and independently substituted with 1-6 groups selected from carboxyl, halogen, C ₁ -C ₆ alkoxy, C ₁ -C ₆ alkyl, and phenyl;

each R ⁷ and R ^{8 is} independently hydrogen, C1-C3 alkyl, C1-C3 alkoxy, phenyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, or 5-7 membered heteroaryl, where alkyl, alkoxy, phenyl, cycloalkyl, heterocyclyl, or heteroaryl represented by R ⁷ or R ⁸ is optionally and independently substituted with 16 groups selected from 5-6 membered heterocyclyl optionally substituted with one or two —F, carboxyl or —CO (OC1-6alkyl), 5- 6-membered heteroaryl, -CN, -OH, O ₁ -C ₃ -alkyl, optionally substituted -NH ₂ or-OH, C ₁ -C ₃ -halogenoalkyl, C ₁ -C ₃ -haloalkoxy, C ₁ -C ₃ - alkoxy -NHCO (C1-Cs-alkyl), -NHCO (C1-Cs-alkoxy), -S (O) 2NRR, -NHS (O) 2NR'R, -NHS (O) 2 (C1-Cs-alkyl) , -NR'R and -C (O) NR'R; each R ⁹ is C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C 1 -C 6 haloalkoxy, or _{C 1 -C 6} alkoxy;

each R ¹⁰ represents a _C1-C3 alkyl optionally substituted with 1-6 groups selected from 5-6 membered heterocyclyl, optionally substituted by one or two atoms -F, carboxy or -CO (OC _1-6 alkyl), C ₃ -C ₆ -cycloalkyl, 5-6-membered heteroaryl, -CN, -OH, -NHCO (C ₁ -C ₃ alkyl), -NHCO (C1-Cz-alkoxy), -S (O) 2NR'R, -NHS ^ NRR, -NHS (O) 2 (C1-Cs-alkyl), -NR'R or -C (O) NR'R; and each R 'and R independently represents hydrogen, methyl, ethyl or propyl; or R 'and R together with the nitrogen atom to which they are attached form a 5-6 membered heterocyclyl; with the proviso that at least one of R ² and R ^{3 is} not hydrogen.

In yet another aspect of the invention, R ¹ in formula (I) is —C (O) NR ⁷ R ⁸ , —CN, phenyl, 56 membered heteroaryl, —C (O) NR'NR'C (O) R ⁹ , -C (O) NR'OR ¹⁰ or C1-C6-alkyl group, where the alkyl group is substituted by 1-3 groups consisting of halogen, C1-C3-alkoxy, -OH, -CN, -NR7R ⁸ , -NR ⁷ COR ^9, 56-membered heteroaryl, and 5-7-membered heterocyclyl, and wherein phenyl and heteroaryl are presented among R ¹ optionally and independently substituted by 1-3 groups selected from halogen, -OH, C1-C3 alkoxy, -CN, -NR7R ⁸ and -CONR ⁷ R ⁸ ;

R ⁶ is hydrogen, C1-C12-alkyl, C1-C4-alkyl-C1-C3-alkoxy- (NR'C1-C6-alkyl) -C1-C3 alkoxy, C1-C4-alkyl-C1-C3-alkoxy- C1-C3 alkoxy, C2-C12 alkenyl, C3-C10 cycloalkyl, 5-6 membered heteroaryl and 5-7 membered heterocyclyl, where alkyl, alkenyl, cycloalkyl, heteroaryl and heterocyclyl are optionally and independently substituted with 1-6 groups selected from carboxyl, halogen, C1-C6 alkoxy, C1-C6 alkyl and phenyl (alternatively, R ⁶ is C1-C12 alkyl, C ₁ -C ₄ alkyl-C ₁ -C ₃ alkoxy- (NR'C ₁ -C ₆ -alkyl) -C ₁ -C ₃ -alkoxy, C ₁ -C ₄ -alkyl-C ₁ -C _3- alkoxy-C ₁ -C ₃ -alkoxy, C ₂ -C ₁₂ - alkenyl, C ₃ C ₁₀ -cycloalkyl, 5-6 membered heteroaryl and 5-7 membered heterocyclyl, where alkyl, alkenyl, cycloalkyl, heteroaryl and heterocyclyl are optionally and independently substituted with 1-6 groups selected from carboxyl, halogen, C1- C ₆ alkoxy, C 1 -C 6 alkyl and phenyl); and each R ⁷ and R ^{8 is} independently hydrogen, C1-C3 alkyl, C1-C3 alkoxy, phenyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, where alkyl, alkoxy, phenyl , cycloalkyl, heterocyclyl or heteroaryl represented by R ⁷ or R ⁸ is optionally and independently substituted with 1-6 groups selected from 5-6 membered heterocyclyl optionally substituted with one or two —F, carboxyl or —CO (OC1-6alkyl) , 5-6 membered heteroaryl, -CN, -OH, _C1-C3 alkyl optionally substituted with -NH ₂ or -OH, C _r C ₃ haloalkyl, _C1-C3 -galogenalkoksi, _C1-C3 alkoxy - NHCO (C ₁ -C ₃ -alkyl), -NHCO (C ₁ -C ₃ -alkoxy), S (O) 2NR'R, -NHS (o) 2NR'R, -NHS (O) 2 (C1-Cz -alkyl), -NR'R and -C (O) NR'R; each R ⁹ is _C1-C6 alkyl, _C1-C6 haloalkyl, _C1-C6 -galogenalkoksi or _C1-C6 alkoxy;

In another aspect, the invention relates to a compound of formula (II)

o (P); or a pharmaceutically acceptable salt thereof, wherein the variables R ¹ , R ² , R ⁴ , R ⁵ and R ⁶ are as defined for formula (I).

In one aspect, the invention relates to a compound of formula (III)

o (w)

- 4037916 or a pharmaceutically acceptable salt thereof, wherein the variables R ¹ , R ³ , R4, R ⁵ and R ⁶ are as defined for formula (I).

In another aspect, the invention relates to a compound of formula (IV)

or a pharmaceutically acceptable salt thereof, wherein variables R4, R ⁵ and R ⁶ are as defined for formula (I).

In another aspect, the invention relates to a compound of formula (V)

or a pharmaceutically acceptable salt thereof, wherein variables R4, R ⁵ and R ⁶ are as defined for formula (I).

In another aspect, the invention relates to a compound of formula (Ia)

or a pharmaceutically acceptable salt thereof; where: R ¹ is -C (O) NR ⁷ R ⁸ , -CN, phenyl, 57-membered heteroaryl, -C (O) NR'NR'C (O) R ⁹ , -C (O) NR'OR ¹⁰ or a C1-C6 alkyl group, where the alkyl group is substituted by 1 to 3 groups consisting of halogen, C1-C3 alkoxy, -OH, -CN, -NR ⁷ R ⁸ , -NR7COR ⁹ , 57-membered heteroaryl and 5 -7-membered heterocyclyl, and wherein phenyl and heteroaryl represented among R ¹ are optionally and independently substituted with 1-3 groups selected from halogen, -OH, C1-C ₃ -alkoxy, -CN, -NR7R ⁸ and -CONR7R ⁸ ; R ² and R ^{3 are} independently selected from hydrogen, halogen, C1-C3-α-alkyl, and C3-C6 cycloalkyl; R4 and R ⁵ are independently selected from hydrogen, halogen, -CN, -CO2R ^9, _C1-C3 alkyl and C1-C ₃ haloalkyl; each R ⁷ and R ^{8 is} independently hydrogen, C1-C3 alkyl, C1-C3 alkoxy, phenyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, or 5-7 membered heteroaryl, where alkyl, alkoxy, phenyl, cycloalkyl, heterocyclyl, or heteroaryl represented by R ⁷ or R ⁸ is optionally and independently substituted with 1-6 groups selected from 5-6 membered heterocyclyl optionally substituted with one or two —F, carboxyl or —CO (OC1-6 alkyl), 5-6 membered heteroaryl, -CN, -OH, C ₁ -C ₃ alkyl optionally substituted with -NH ₂ or -OH, C1-C ₃ -haloalkal, C1-C ₃ -haloalkoxy, C ₁ -C ₃ alkoxy -NHCO (C ₁ -C ₃ -alkyl), -NHCO (C ₁ -C ₃ -alkoxy), -S (O) ₂ NR'R, -NHS (O) ₂ NR'R, -NHS (O ) ₂ (C ₁ -C ₃ alkyl), -NR'R and -C (O) NR'R; each R ⁹ is C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 haloalkoxy or C1-C6 alkoxy; each R ¹⁰ is C1-C3 alkyl optionally substituted with 1-6 groups selected from 5-6 membered heterocyclyl optionally substituted with one or two —F, carboxyl or —CO (OC _1-6 alkyl), C ₃ -C ₆ -cycloalkyl, 5-6-membered heteroaryl, -CN, -OH, -NHCO (C1-Cs-αalkyl), -NHCO (C1-Cs-alkoxy), -S (O) 2NR'R, -NHS (O) 2NRR, -NHS (O) 2 (Ci-C3 alkyl), -NR'R, or -C (O) NR'R; and each R 'and R independently represents hydrogen, methyl, ethyl or propyl; or R 'and R together with the nitrogen atom to which they are attached form a 5-6 membered heterocyclyl; with the proviso that at least one of R ² and R ^{3 is} not hydrogen.

In another aspect, the invention relates to a compound of formula (Ia)

or a pharmaceutically acceptable salt thereof; where: R ¹ is -C (O) NR ⁷ R ⁸ , -CN, phenyl, 56-membered heteroaryl, -C (O) NR'NR'C (O) R ⁹ , -C (O) NR'OR ¹⁰ or C ₁ -C ₆ -alkyl group, where alkyl

- 5 037916 the group is substituted by 1-3 groups consisting of halogen, C ₁ -C ₃ -alkoxy, -OH, -CN, -NR ⁷ R ⁸ , -NR ⁷ COR ⁹ , 56-membered heteroaryl and 5-7-membered heterocyclyl, and where phenyl and heteroaryl, represented among R ¹ , are optionally and independently substituted by 1 to 3 groups selected from halogen, —OH, C1-C3 alkoxy, —CN, —NR7R ⁸ and —CONR7R ⁸ ; R ² and R ^{3 are} independently selected from hydrogen, halogen, C ₁ -C ₃ -alkyl and C _{3 -C 6} cycloalkyl; R4 and R ^{5 are} independently selected from hydrogen, halogen, —CN, —CO2R ⁹ , C1-C3 alkyl, and C1-C3 haloalkyl; each R ⁷ and R ^{8 is} independently hydrogen, _{C 1} -C ₃ alkyl, C _{1 -C 3} alkoxy, phenyl, C ₃ -C ₆ cycloalkyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, where alkyl, alkoxy, phenyl, cycloalkyl, heterocyclyl or heteroaryl represented among R ⁷ or R ⁸ is optionally and independently substituted with 1-6 groups selected from 5-6 membered heterocyclyl optionally substituted with one or two atoms -F, carboxyl or -CO (OC1-6alkyl), 5-6 membered heteroaryl, -CN, -OH, C1-C3 alkyl optionally substituted with -NH2 or -OH, C1-C3 haloalkyl, C1-C3 haloalkoxy, C1C3 alkoxy -NHCO (C1-Cs-alkyl), -NHCO (C1-Cs-alkoxy), -S (O) 2NR'R, -NHS (O) 2NRR, -NHS (O) 2 (CrC3alkyl), -NR'R and -C (O) NR'R; each R ⁹ is _{C 1} -C ₆ alkyl, C _{1 -C 6} haloalkyl, C ₁ -C ₆ haloalkoxy or C 1 -C 6 alkoxy; each R ¹⁰ is C ₁ -C ₃ alkyl optionally substituted with 1-6 groups selected from 5-6 membered heterocyclyl, optionally substituted with one or two —F, carboxyl or —CO (OC _1-6 alkyl), C ₃ -C ₆ -cycloalkyl, 5-6-membered heteroaryl, -CN, -OH, -NHCO (C ₁ -C ₃ -alkyl), -NHCO (C ₁ -C ₃ -alkoxy), -S (O) ₂ NR'R, -NHS (O) ₂ NR'R, -NHS (O) ₂ (C1-C ₃ alkyl), -NR'R or -C (O) NR'R; and each R 'and R independently represents hydrogen, methyl, ethyl or propyl; or R 'and R together with the nitrogen atom to which they are attached form a 5-6 membered heterocyclyl; with the proviso that at least one of R ² and R ^{3 is} not hydrogen.

In another aspect, the invention relates to a compound of formula (IIa)

or a pharmaceutically acceptable salt thereof, wherein the variables R ¹ , R ² , R ⁴ and R ⁵ are as defined for formula (Ia).

In one aspect, the invention relates to a compound of formula (IIIa)

or a pharmaceutically acceptable salt thereof, wherein the variables R ¹ , R ³ , R ⁴ and R ⁵ are as defined for formula (Ia).

In another aspect, the invention relates to a compound of formula (IVa)

or a pharmaceutically acceptable salt thereof, wherein the variables R ⁴ and R ⁵ are as defined for formula (Ia).

In another aspect, the invention relates to a compound of formula (Va)

or a pharmaceutically acceptable salt thereof, where the variables R4 and R ⁵ are as defined for formula (Ia).

In one aspect of the inventions, for formulas (I), (Ia), (II) or (IIa), R ² is C1-C ₃ alkyl. In yet another aspect of the invention, for formulas (I), (Ia), (II) or (IIa), R ² is methyl.

In one aspect of the invention, for formula (I), (Ia), (III) and (IIIa), R ³ is C1-C3 alkyl. In yet another aspect of the invention, for formula (I), (Ia), (III) and (IIIa), R ³ is methyl.

- 6 037916

In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), R ^{1 is} selected from oxadiazole, —C (O) NHNHC (O) (C1- Cz-alkyl), -CH2NH2, -CH2NHCO (C1-Cz-α-alkoxy), -CH2NHCO (Ci-C3 alkyl) or -CH2NHCO (C1-C3-haloalkyl), where oxadiazole R ^{1 is} optionally substituted with -OH, C1-C ₃ alkoxy ,

-NR ⁷ R ⁸ or -CONR ⁷ R ⁸ . In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), R ^{1 is} selected from —CH2NH2,

In yet another aspect of the invention, for formula (I), (Ia), (IIa), (III) and (IIIa), R ¹ is CN,

where R ¹¹ represents hydrogen or -C (O) NH2. In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), R ¹ is —CN or —C (O) NH2. In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), R ¹ is —CN. In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), R ¹ is —C (O) NH2. In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), R ¹ is —C (O) NR ⁷ R ⁸ . In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), when R ¹ is —C (O) NR ⁷ R ⁸ , R ⁷ is is hydrogen and R ⁸ is 1) phenyl optionally substituted with C1-C3-αalkyl or C1-C3-alkyl-NH2, 2) C1-C3-alkyl or 3) C1-C3-alkoxy, where each is alkyl or alkoxy from and R ⁸ is optionally independently substituted with _C3-C6 cycloalkyl, -CN, -OH, -NH2, -SO2NH2, -NHSO2NH2, -C (O) NH2, -NHC (O) (C ₁ -C ₃ -alkyl), pyrazinyl, oxytanyl, oxazolyl, or pyrrolidinyl optionally substituted with one or more carboxyl, fluorine, or —C (O) O (C ₁ -C ₆ alkyl). In one aspect of the invention, for formula (I), (Ia), (II), (IIa), (III) and (IIIa), when R ¹ is —C (O) NR ⁷ R ⁸ , R ⁷ is is hydrogen and R ^{8 is} selected from the group consisting of

-CH2CN and -CH2OH.

In one aspect of the invention, for any of formulas (I), (II), (III), (IV), or (V), R ⁶ is C ₁ -C ₁₂ alkyl. In one aspect of the invention, for any of formulas (I), (II), (III), (IV) or (V), R ⁶ is ethyl, isopropyl, 2-butyl, or isopentyl. In one aspect of the invention, for any of formulas (I), (II), (III), (IV), or (V), R ⁶ is isopropyl. In one aspect of the invention, for any of formulas (I), (II), (III), (IV), or (V), R ⁶ is C ₁ -C ₄ -αalkyl-OC (O) - (NHC ₁ C ₆ -alkyl) -C (O) C ₁ -C ₃ -alkoxy, C ₁ -C ₄ -alkyl-OC (O) -C ₁ -C ₄ -alkyl or C ₁ -C ₄ -alkyl-OC (O ) -C ₁ -C ₃ -alkoxy. In one aspect of the invention, for any of formulas (I), (II), (III), (IV), or (V), R ^{6 is} selected from the group consisting of

- 7 037916

methyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, or n-nonyl.

In one aspect of the invention, for any of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), R ⁴ and R ⁵ independently represent H, methyl or fluoro. In yet another aspect of the invention, for any of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), one of R ⁴ and R ⁵ is hydrogen and the other is fluoro. In yet another aspect of the invention, for any of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), R ⁴ is fluorine and R ⁵ is hydrogen. In yet another aspect of the invention, for any of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), R ⁴ is hydrogen and R ⁵ is fluoro. In yet another aspect of the invention, for any of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), R ⁴ and R ^{5 are} both hydrogen. In yet another aspect of the invention, for any of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), R ⁴ and R ^{5 are} both fluoro.

Any embodiment described herein can be combined with any other suitable embodiment described herein to provide additional embodiments. For example, when one of the embodiments individually or collectively describes possible groups for R ¹ , and in another embodiment describes possible groups for R ² , it is contemplated that these embodiments can be combined to provide a further embodiment in which any of the possible groups for R ¹ with any of the possible groups for R ² . Likewise, the invention encompasses any embodiments relating individually to R ¹ , R ² , R ³ , R ⁴ , R ^5, and R ⁶ , in combination with any individual embodiments relating to each of the remaining variables.

The compounds of formulas (I), (II), (III), (IV) and (V) and their pharmaceutically acceptable salts have an effective spectrum of beta-lactamase inhibition and are suitable for oral administration. Compounds of formulas (Ia), (IIa), (IIIa), (IVa) and (Va) and their pharmaceutically acceptable salts have an effective spectrum of beta-lactamase inhibition and are suitable for intravenous, intraperitoneal, intramuscular or subcutaneous administration, for example, intravenous administration. As such, the compounds of formulas (Ia), (IIa), (IIIa), (IVa) and (Va) and their pharmaceutically acceptable salts can be successfully used when the patient cannot take antibiotics orally, for example, in a hospital environment, conditions of paragraph emergency care or nursing home conditions. Once the patient has improved enough to take oral antibiotics, treatment can be changed so that the compound of formulas (I), (II), (III), (IV) and (V) or a pharmaceutically acceptable salt thereof is orally administered to the patient. In addition, the compounds of formulas (Ia), (IIa), (IIIa), (IVa), (Va), (I), (II), (III), (IV) and (V) and their pharmaceutically acceptable salts can have beneficial, effective, metabolic, toxicological and / or pharmacodynamic properties.

One aspect of the invention includes a compound according to one of the examples, or a pharmaceutically acceptable salt thereof, namely:

Compound name (I) -ethyl- 2 - ((2 S, 5H) -2-carbamoyl-3 methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yloxy) -2-fluoroacetate ( 8) -ethyl- 2 - ((28.5H) -2-carbamoyl-3methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yloxy) -2-fluoroacetate (2S) - {[ (2 S, 5H) -2-carbamoyl-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3 en-6-yl] oxy} (fluoro) ethanoic acid lithium salt (2R) - { [(2 S, 5H) -2-carbamoyl-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3 en-6-yl] oxy} (fluoro) ethanic acid lithium salt

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five h ₂ n ^A A Μ F о {[(2 S, 5H) -2-carbamoyl-3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy} (fluoro) acetic acid lithium salt 6 7 eight nine 10 AND 12 T I I I I T T Μ Μ ΓΌ Μ ΓΌ Μ ΓΌ z z z z z z z About A o. ) = ° O. <= O o. <= ° o. Ao o. Ao o. Ao ϊί> ϊί> ϊί> tF o 'o' o 'o' o 'o' o ' 0 ^ 2 '^ s s k ethyl- {[(2 S, 5H) -2-carbamoyl-3-methyl7-oxo-1, 6-diazabicyclo [3.2.1] oct-3 en-6-yl] oxy} (fluoro) acetate ethyl- {[(2 S, 5H) -2-carbamoyl-3-methyl7-oxo-1, 6-diazabicyclo [3.2.1] oct-3 en-6-yl] oxy} (difluoro) acetate {[(2 S, 5H) -2-carbamoyl-3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy} (difluoro) acetic acid lithium salt ethyl- {[(2 S, 5H) -2-carbamoyl-3-methyl7-oxo-1, 6-diazabicyclo [3.2.1] oct-3 en-6-yl] oxy} acetate {[(2 S, 5H) -2-carbamoyl-3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy (acetic acid lithium salt 2- {[(2 S, 5H) -2-carbamoyl-3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy (-2-fluoropropane acid lithium salt propan-2-yl- (2I.) - [[(2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy} (fluoro) etanoate 13 A o ' Ο ^ Ή _ ^ o propan-2-yl- (28) - [[(2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy} (fluoro) ethanoate fourteen o I η ₂ ν ^Λ Α MF ^? Ay 2,4-dimethylpentan-3-yl- (28) - {[(2 S, 5H) -2-carbamoyl-3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en- 6-yl] oxy} (fluorine) etanoate fifteen I z About Ao ϊ> o ' 2,4-dimethylpentane-3-yl- (2H) - {[(2 S, 5H) -2-carbamoyl-3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy} (fluoro) ethanoate sixteen nLA M> —h, ^{F 0} ° ^ ° o tetrahydro-2H-pyran-4-yl - {[(28.5K) 2-carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy} (fluoro) acetate 17 h _! n ^A A MF J --- N / ^{0 0} IT ⁰ o ^{4 4} o ^ 2-methoxy- {[(2 S, 5H) -2-carbamoyl-3 methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-ene-6yl] oxy} (fluoro) acetate 18 h ₂ n ^A A MFA — N r ⁰ oA ₀ θ 'o 2-methoxy- (2K) - {[(28.5K) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy} (fluoro) ethanoate nineteen H ₂ N ^A A MF A — N ₄ J ° ⁰ ) A o 'O' 2-MeTOKCH- (2S) - {[(2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy} (fluoro) ethanoate twenty H ₂ N ^A A MF J --- N · ⁰ ° ^ Г ° o ' (2R) - (S) -BTop-6yTmi-2 - (((2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate

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21 aA М F J - NJ ⁰ ° / 7 ° 0} ⁴ (2S) - (S) -BTop-6yTHn-2 - (((2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 22 ⁰ 1 Αλ r ⁰ '°> ° ⁰ ) ⁴ (2R) - (R) -BTop-6yTmi-2 - (((2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 23 ⁰ 1 aA jA j ⁰ ° / Г ^{0 0} ) ⁴ (2S) - (R) -BTop-6yran-2 - (((2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 24 j p Pi О = / о z T (R) -neHTa _H -3 -yl-2 - ((2 S, 5R) -2carbamoyl-3 -methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yloxy) -2- fluoroacetate 25 JA Ή F A— ^N j ⁰ ° / Г ⁰ ο (S) -neHTaH-3-mi-2 - ((2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yloxy) -2-fluoroacetate 26 A _r A - ^N i ^{0 0} / r ⁰ ^ 0 3Tmi-2 - (((2S, 5R) -2- (2- acetylhydrazinecarbonyl) -3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl) oxy) -2-fluoroacetate 27 r A. A— ^N , / ⁰ 'O'y-OH 0 2 - (((2S, 5R) -2- (2- acetylhydrazinecarbonyl) -3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl) oxy) -2-fluoroacetic acid lithium salt 28 d TZ About Ao ID- o ' A ™ (R) -2 - ((2S, 5R) -2- (4- (aminomethyl) phenylcarbamoyl) -3 methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-ene-6yloxy) -2-fluoroacetic acid TFA salt 29 ⁰ 1 (PA ⁰ ° / G ⁰ ^ o ⁷ ethyl 2-fluoro-2 - (((2 S, 5R) -3 -methyl-7 oxo-2 - ((pyrazine-2ylmethyl) carbamoyl) -1.6 diazabicyclo [3.2.1] oct-3-en-6yl ) hydroxy) acetate thirty ⁰ 1 AG ^ A ^N J ⁰ oDri o ⁷ 2-fluoro-2 - (((2 S, 5R) -3-methyl-7-oxo-2 ((pyrazin-2-ylmethyl) carbamoyl) -1,6 diazabicyclo [3.2.1] oct-3-en-6yl ) hydroxy) acetic acid lithium salt 31 0 A η ₂ ν γ ηρ Μ F D — Ν? ° ° / Γ ⁰ ^ 0 (2R) -3thh- 2 - (((2 S, 5 R) -2-carbamoyl-4methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl) oxy) -2-fluoroacetate 32 0 l η ₂ ν γ ίΓ Μ F J --- Ν / ⁰ '° / Γ ⁰ ^ ο ⁷ (28) -ethyl- 2 - (((28,5R) -2-Kap6aMonn-4- methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl) oxy) -2-fluoroacetate 33 0 η ₂ ν γ γ Μ F J --- Ν _χ ⁰ θ '^ Ρ-ΟΗ ο ⁷ (2R) -2 - (((2S, 5R) -2- _K ap6aMomi-4methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetic acid lithium salt 34 0 Η ₂ Ν [Μ _F J --- Ν _χ J ⁰ ° ^ y-on ο ⁷ (2S) -2 - (((2S, 5R) -2- _K ap6aMomi-4methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetic acid lithium salt

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35 0 A η ₂ ν ρ ηρ Μ F J — ν ί- α ο ^ ο ° 7 ^ (2R) -H3onponmi-2 - (((2S, 5R) -2carbamoyl-4-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 36 0 l η ₂ ν ρ ηρ Μ F λ --- ^Ν / ° οΑ ο о о К (2 8) -isopropyl-2 - (((2 S, 5R) -2 carbamoyl-4-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 37 nD Μ F λ— Ν F ⁰ '° Ύο ο X (2R) -isoπpoπyl-2 - (((2S, 5R) -2carbamoyl-3-cyclopropyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-ene-6yl) oxy) -2-fluoroacetate 38 nL Μ F J --- Ν / ⁰ '° / Г ⁰ ο 7 ^ (2S) -isoπpoπyl-2 - (((2S, 5R) -2carbamoyl-3-cyclopropyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 39 ηΧΧ Μ F J --- Ν ⁰ ° Ίτ ° ^ 0 (2R) -ethyl- 2 - (((2S, 5R) -2-κapamoyl-3cyclopropyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 40 ηΧΧ Μ F Λ— ^Ν / ^{0 0} / Γ ⁰ ^ 0 (28) -ethyl- 2 - (((2S, 5R) -2-κapbamoyl-3cyclopropyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2-fluoroacetate 41 Л Μ F J --- Ν _χ ⁰ 0 (2R) -2 - (((2S, 5R) -2-κapbamoyl-3cyclopropyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-6yl) oxy) -2-fluoroacetic acid 42 J --- Ν _χ / ° ° ΧΟΗ Ο (2R) -2 - (((2S, 5R) -2- _K ap6aMo ™ -3-cyclopropyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2 -fluoroacetic acid lithium salt 43 l tX F ⁰ А-4у, ₀ Н Lithium salt 44 ⁰ 1 n ₂ ЛД> 1 ° ° / Г ° Л ο (1-isopropyl-2-methylpropyl) 2 [[(2 S, 5R) -2-κapbamoyl-3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy] - 2,2-d ifluoro-acetate 45 YES Μ F J ---- ^Ν 4 'ο ° Χ ^ ° \ __ 0 - <· \ oktyl- (2R) -2 - [[(2S, 5R) -2-κapbamoyl- 3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoro-acetate 46 ⁰ 1 Η ₂ Α Α dX g ⁰ ° Ατ ° \ 0 methyl- (2R) -2 - [[(2S, 5R) -2-κapbamoyl- 3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoro-acetate 47 ⁰ 1 dA dX? ⁰ ° Γ ° 0 allyl- (2R) -2- [[(2 S, 5R) -2-κapbamoyl3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoro-acetate 48 ⁰ 1 η ₂ αΑ Μ F A --- ^N s' ⁰ ° l ^ ° o πpoπyl- (2R) -2 - [[(2S, 5R) -2-κapbamoyl- 3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoro-acetate

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49 ⁰ 1 Η ₂ Ν ^Λ Α jA, г ^{0 0} 0 A H3o6yTnn- (2R) -2 - [[(2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy] -2-fluoroacetate fifty ⁰ 1 η / Λ jA f ⁰ о butyl- (2R) -2 - [[(2S, 5R) -2-κapbamoyl3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-ene-6yl] oxy] -2-fluoroacetate 51 n / a M F J ---- N ί- Ο O neHTHn- (2R) -2 - [[(2S, 5R) -2-Kap6aMonn- 3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoro-acetate 52 _H2 AA M F J --- N? ° ° Ίτ ° - \ o reKcnn- (2R) -2- [[(2 S, 5R) -2-κapbamoyl- 3-methyl-7-okco-l, 6- diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoro-acetate 53 η ₂ ν ^Λ Α Μ F A ~ N ₄ F ⁰ o- \ ₀ "Guy o / 1 -isopropoxycarbonyloxyethyl (2R) -2 - [[(2S, 5 R) -2-carbamoyl-3methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy] -2-fluoro -acetate 54 Η ₂ Ν ^Λ Α MF E ---- N _s .- ° ^O "^ T ° ^ A e o (2R) -6eH3Hn-2 - (((2S, 5R) -2-Kap6aMonn- 3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6- yl) oxy) -2-fluoroacetate 55 o H ₂ N— # Ao 1 ν-Λ MF Λ --- N ₄ J o 0 \ OH 0 2 - [[(2S, 5R) -2- (5-Kap6aMonn-l, 3,4oxadiazol-2-yl) -3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene- 6yl] oxy] -2-fluoroacetic acid lithium salt 56 ^{0 H} jA J ⁰ O'y-OH 0 (2S) -2XTop-2 - [[(2S, 5R) -3-Meran-7oxo-2- (2- sulfamoylethoxycarbamoyl) -1,6 diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt 57 O_ o I A — N ₄ J ^{0 O} ^ y-OH 0 (2S) -2XTop-2 - [[(2S, 5R) -3-Meran-7oxo-2- (2- sulfamoylethylcarbamoyl) -1,6 diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt 58 AA MF J --- NF ° ° Ar ° ^ o H 0 ⁰ A ^N yA _ 0 __ / o ^ \ 3Tmi-2 - [[(2R) -2 - [[(2S, 5R) -2carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy] -2 -fluorine- acetyl] oxymethoxycarbonylamino] 3-methylbutanoate 59 ⁰ 1 YX, A — N ₄ J ^{0 0} yO ^. O T _P er-6yTmi- (2S) -2 - ((((2S, 5R) -6 - ((S) -2ethoxy-1-fluoro-2-oxoethoxy) -3 methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-ene-2carboxamido) oxy) methyl) pyrrolidi n-1-carboxylate 60 A 0 I Αχ · A - nj ^{0 0} / Γ ° \ ^ 0 ethyl- (2 8) -2-fluoro-2 - (((2 S, 5R) -3 -methyl7-oxo-2 - ((((8) -pyrrolidin-2yl) methoxy) carbamoyl) -1,6 diazabicyclo [ 3.2.1] oct-3-ene-6yl) oxy) acetate TFA salt 61 A ", A ^ _r ⁰ ° ΑΆ 0 3THn- (2S) -2 - (((2S, 5R) -2 - ((((S) -4,4 difluoropyrrolidine-2- yl) methoxy) carbamoyl) -3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl) oxy) -2-fluoroacetate TFA salt

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62 o 1 A — N _s / ⁰ ° y-on 0 (28) -2-fluoro-2 - [[(28.5K) -3-methyl-7oxo-2 - [[(28) -pyrrolidine-2- yl] methoxycarbamoyl] -1,6 diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid TFA salt 63 64 65 66 67 Α θ, 'A · Α — Ν ₄ / Η ₂ Ν ^Α Α Μ F + --- Ν? ° / Ο Υ'ν η ₂ ν ^Α Α Μ F A — Ν? ⁰ ° ^ 7r ° ο Ul-l ⁰ O'-y-on ^{0 Η} W; ⁰ ° y-he 0 (2S) -2 - (((2S, 5R) -2 - ((((S) -4,4 difluoropyrrolidin-2yl) methoxy) carbamoyl) -3-methyl-7 oxo-1,6-diazabicyclo [3.2.1 ] oct-3-en6-yl) oxy) -2-fluoroacetic acid TFA salt [(2R) -2 - [[(2S, 5R) -2- _K ap6aMOwi-3methyl-7-oxo-1,6-diazabicyclo [ 3.2.1] oct-3-en-6yl] oxy] -2-fluoro-acetyl] oxymethyl2, 2-dimethylpropanoate indan-5-yl- (2R) -2 - [[(2S, 5R) -2carbamoyl-3 - methyl 7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl] oxy] -2-fluoroacetate (28) -2-fluoro-2 - [[(28 ^) - 3-methyl-2 (oxetan-3-ylcarbamoyl) -7-oxo-1,6 diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt (2S) -24top-2 - [[(2S, 5R) - 2- [2- (methanesulfonamido) ethylcarbamoyl] -3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt 68 ⁰ j ⁰ ° f ^OH 0 (28) -2-fluoro-2 - [[(28 ^) - 3-methyl-2- (oxazol-2-ylmethylcarbamoyl) -7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy] acetic acid lithium salt 69 ⁰ ° A ^ OH 0 (28) -2-fluoro-2 - [[(28 ^) - 3-methyl-7- oxo-2- (pyrazine-2- ilmethylcarbamoyl) -1.6- d isabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt 70 A ⁰ 1 ^ 'ά, A— n _s j ⁰ θ ^ Α ^ ΟΗ 0 (2S) -2 - [[(2S, 5R) -2- (cyclopropylmethoxycarbamoyl) -Zmethyl-7-oxo-1,6- d isabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoroacetic acid lithium salt 71 О O 1 A – n _x j о O ^ A ^ OH 0 (2S) -2 - [[(2S, 5R) -2 - [(3-aMHHO-3-OKCOpropyl) carbamoyl] -3 -methyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-ene -6yl] oxy] -2-fluoroacetic acid lithium salt 72 O = \ Jl О I A - n ₄ j ^{0 o} / r he 0 (2S) -2 ^ Top-2 - [[(2S, 5R) -3-MeTHn-7oxo-2 - [(5-oxopyrrolidine-2yl) methoxycarbamoyl] -1.6d isabicyclo [3.2.1] oct-3 - en-6yl] oxy] acetic acid lithium salt 73 o = \ Ί О I = A — n ₄ j ⁰ θ ^ Α ^ ΟΗ 0 (2S) -2 ^ Top-2 - [[(2S, 5R) -3-Me _T Hn-7oxo-2- [2- (5-oxopyrrolidin-2yl) ethylcarbamoyl] -1,6-d iazabicyclo [3.2. 1] oct-3-en-6yl] oxy] acetic acid lithium salt 74 A p O-U) IZ O. Ao M- o Οΐ - ^^ Ή o I (28) -2-fluoro-2 - [[(28 ^) - 3-methyl-7- oxo-2- (3- sulfamoylpropylcarbamoyl) -1.6d isabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt

- 13 037916

75 θΑ ^ ΛΑ ^H2f T n A A — ν ₄ j ⁰ O'y-OH 0 (2 8) -2-φτορ-2- [[(2 S, 5R) -3 -methyl-7okco-2- [2- (sulfamoylamino) ethylcarbamoyl] 1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt 76 77 78 79 80 81 7 7 g "> A Y, = <=. = /. = “S v- (Г Μ ϊ> A 1 AA θ 'O ^z O ^z _n Α-η A У. к km А О Я oA ^t ο-Α ^ ή oA ^m о ¹ V \ \ A i OOO d ( ( ¹ § (2S) -2 - [[(2S, 5R) -2 - [( _T peTbutoxycarbonylamino) methyl] -3 methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy ] -2-fluoroacetic acid lithium salt (2S) -2 - [[(2S, 5R) -2- (aminomethyl) -3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3- en-6yl] oxy] -2-fluoro-acetic acid (2 S) -2- [[(2 S, 5R) -2- (acetamide omethyl) 3-methyl-7-oxo-1,6-diazabicyclo [3.2 .1] oct-3-ene-6yl] oxy] -2-fluoroacetic acid lithium salt (2 8) -2-fluoro-2- [[(2 S, 5R) -3 -methyl-7oxo-2- [[(2,2,2-trifluoroacetyl) amino] methyl] -1,6 diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt of etyl- (2S) -2-φτop-2- [[(2S, 5R) -2 (cyanomethylcarbamoyl) -3 -methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy] acetate ethyl- (2R) -2-φτop -2 - [[(2S, 5R) -2 (cyanomethylcarbamoyl) -3-methyl-7 oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy] acetate 82 ⁰ 1 aAA J - N _s J o ° A0H 0 (2S) -2 ^ Top-2 - [[(2S, 5R) -2- (cyanomethylcarbamoyl) -3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy] acetic acid lithium salt 83 ° "aLA ^H2 % ⁿ A _F A — nj o __ o ethyl- (2S) -2-φτop-2 - [[(2S, 5R) -2 (cyanomethylcarbamoyl) -3 -methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy ]acetate 84 % άΛα ^H О _F A ~ ⁿ ^ a ° о ethyl- (2R) -2-φτop-2 - [[(2S, 5R) -2 (cyanomethylcarbamoyl) -3 -methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy ] acetate 85 ⁰ 1 HO ^ N ^ 'Al ^H P _F ^{0 0} rV 0 ethyl- (2S) -2-φτop-2 - [[(2S, 5R) -2 (hydroxymethylcarbamoyl) -3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3 en-6-yl] hydroxy] acetate 86 ⁰ 1 A _F A ^ ^N 4 - ' ⁰ 0 ethyl- (2R) -2-φτop-2 - [[(2S, 5R) -2 (hydroxymethylcarbamoyl) -3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3 en-6-yl] hydroxy] acetate 87 ⁰ 1 A _r A - N ₄ J ο Ο'γ-ΟΗ 0 (2S) -2 ^ Top-2 - [[(2S, 5R) -2- (hydroxymethylcarbamoyl) -3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3 en-6-yl] oxy] acetic acid lithium salt 88 0 0 I AMA ^{n n} 0 _F A – n ₄ j ο θ'γ-ΟΗ 0 (2S) -24top-2 - [[(2S, 5R) -2- (acetamidomethylcarbamoyl) -3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3 en-6-yl] oxy] acetic acid lithium salt

- 14 037916

89 % o and 1 H ₂ N ' ^S ' N ^ N ^ '''f4 ^{H N} Y F Y - m _h J ⁰ O (28) -2-fluoro-2 - [[(28.5K) -3-methyl-7oxo-2- [(sulfamoylamino) methylcarbamoyl] -1, 6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy] acetic acid lithium salt 90 О Η ₂ Ν М F _C 0 ethyl 2 - (((2 S, 5 R) -2-carbamoyl-4methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2,2-difluoroacetate 91 o H ₂ N _F o 2 - (((28.5H) -2-carbamoyl-4-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en- 6-yl) oxy) -2,2-difluoroacetic acid lithium salt 92 0 J --- N _ o / o ⁰ if 0 ethyl 2 - (((2 S, 5 R) -2-carbamoyl-4methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-ene-6yl) oxy) acetate 93 0 h ₂ n ο ^ ^Ν ο-γ ^ΟΗ 0 2 - (((28.5H) -2-carbamoyl-4-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl) oxy) acetic acid lithium salt 94 ΟΓ F E --- N J. o / o 0 ethyl- (2L) -2 - (((2 S, 5K) -2-cyano-4methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-ene-6yl) oxy) -2-fluoroacetate 95 ^Ы ГГГ F o ^ ~ ^N 'oV ^H 0 (2K) -2 - (((28.5K) -2-cyano-4-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl) oxy) -2-fluoroacetic acid lithium salt 96 0 J ^N s ^{0 0} iT] O 1 isopropyl-2 - (((2 S, 5H) -2-carbamoyl4-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-ene-6yl) oxy) acetate

Compounds of the invention include compounds of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), (Va) in free base form ( uncharged), as well as their pharmaceutically acceptable salts.

Alkyl — As used herein, the term alkyl refers to straight and branched chain saturated hydrocarbon radicals having the indicated number of carbon atoms. References to individual alkyl groups such as propyl characterize only the straight chain version, and references to individual branched chain alkyl groups such as isopropyl and 3-pentyl characterize only the branched chain version. In one aspect, alkyl is methyl.

Halogen — As used herein, the term halogen is intended to include fluorine, chlorine, bromine and iodine. In one aspect, halogen can refer to fluorine, chlorine and bromine. In yet another aspect, halogen can be fluorine or chlorine. In yet another aspect, the halo can be fluorine. In yet another aspect, halogen can refer to chlorine.

Haloalkyl - as used herein, is an alkyl moiety as defined above substituted with one or more halogen atoms. In one aspect, the haloalkyl can be —CHF2, —CH2F, or —CF ₃ .

Cycloalkyl — In one aspect, cycloalkyl refers to a saturated monocyclic carbon ring, one or more —CH2— groups of which may optionally be replaced with an appropriate number of —C (O) - groups. Illustrative examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, and cyclopentenyl. In one aspect, the 3-5 membered carbocyclyl can be cyclopropyl.

5-7 membered heterocyclyl - The term 5-7 membered heterocyclyl refers to a saturated or partially saturated non-aromatic monocyclic ring containing from 5 to 7 ring atoms, at least one of which is selected from nitrogen, sulfur and oxygen, and the -CH2 group - which may optionally be replaced by a -C (O) - group. Similarly, 5-6 membered heterocyclyl refers to a saturated or partially saturated non-aromatic monocyclic ring containing from 5 to 6 ring atoms, at least one of which is selected from nitrogen, sulfur and oxygen, and the group

- 15 037916

CH2- of which may optionally be replaced with a —C (O) - group. Unless otherwise indicated, 5-7 membered heterocyclyl and 5-6 membered heterocyclyl groups can be bonded with carbon or nitrogen. Ring nitrogen atoms can optionally be oxidized to form an N-oxide. Ring sulfur atoms can optionally be oxidized to form S-oxides or sulfones. Illustrative examples of the 5-7 membered heterocyclyl and the 5-6 membered heterocyclyl include, but are not limited to, azetidinyl, dioxidotetrahydrothiophenyl, 2,4-dioxoimidazolidinyl, 3,5-dioxopiperidinyl, furanyl, imidazolyl, oxothiazolyl, isofoxoliniyl, oxolidinyl , 3-oxo-1-piperazinyl, 2-oxopyrrolidinyl, 2-oxotetrahydrofuranyl, oxo1,3-thiazolidinyl, piperazinyl, piperidyl, 2H-pyranyl, pyrazolyl, pyridinyl, pyrrolyl, pyrrolidinyl, pyrimidinyl, pyrazinyl, pyrrolidinyl, pyrimidinyl, pyrazinyl, 4 , tetrahydrofuranyl, tetrahydropyranyl, thiazolyl, 1,3,4-thiadiazolyl, thiazolidinyl, thiomorpholinyl, thiophenyl, 4H-1,2,4-triazolyl, pyridine N-oxidyl, tetrazolyl, oxadiazolyl, triazolyl, pyrazinyl, triazinyl, and homopipiperidinyl. In one embodiment, the terms 5-7 membered heterocyclyl and 5-6 membered heterocyclyl include 5-7 or 5-6 membered siderophores containing at least one heteroatom.

5- or 6-membered heteroaryl - The term 5-6-membered heteroaryl refers to a monocyclic aromatic heterocyclyl ring containing 5 or 6 ring atoms, at least one of which is selected from nitrogen, sulfur and oxygen. Unless otherwise indicated, 5-6 membered heteroaryl groups can be bonded with carbon or nitrogen. Ring nitrogen atoms can optionally be oxidized to form an N-oxide. Ring sulfur atoms can optionally be oxidized to form S-oxides. Illustrative examples of 5-6 membered heteroaryl include furanyl, imidazolyl, isothiazolyl, isoxazole, oxazolyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, pyridinyl, pyrrolyl, tetrazolyl, thiadiazolyl, thiazolyl, thiophenyl, and triazolyl.

Optionally Substituted — As used herein, the expression optionally substituted indicates that the substitution is optional and, therefore, the specified group may be substituted or unsubstituted. If substitution is required, the corresponding number of hydrogen atoms in the specified group can be replaced by a selection of these substituents, provided that the normal valence of the atoms in a particular substituent is not exceeded and that the substitution results in a stable compound.

In one aspect, when a particular group is indicated as optionally substituted with one or more substituents, that particular group may be unsubstituted. In yet another aspect, a particular group may carry one substituent. In yet another aspect, a particular substituent may carry two substituents. In yet another aspect, a particular group may carry three substituents. In another aspect, a particular group may carry four substituents. In an additional aspect, a particular group may carry one or two substituents. In yet another additional aspect, a particular group may be unsubstituted or may carry one or two substituents.

Pharmaceutically acceptable - in the context of this document, the expression pharmaceutically acceptable refers to such compounds, substances, compositions and / or dosage forms that are medically suitable for use involving contact with human and animal tissues, without undue toxicity, irritation, allergic reaction or other problems or complications consistent with a reasonable risk / benefit ratio.

Prodrug — As used herein, the term prodrug refers to a chemically modified version of a pharmacologically active agent that undergoes in vivo conversion to release the active drug. (See Rautio, J., et al., Prodrugs: Design and Clinical Applications, Nature Rev., vol. 7, page 255 (March 2008)). In the present invention, prodrugs are used to render active beta-lactamase inhibitor compounds orally bioavailable after absorption from the gastrointestinal tract.

Effective Amount — As used herein, the expression “effective amount” means an amount of a compound or composition sufficient to significantly and positively alter the symptoms and / or conditions to be treated (eg, to provide a positive clinical response). The effective amount of active ingredient for use in the pharmaceutical composition will vary depending on the particular condition to be treated, the severity of the condition, the duration of treatment, the nature of the concomitant therapy, the particular active ingredient (s) used, the particular pharmaceutically acceptable excipient (s) / carrier (s) used. ) and similar factors within the knowledge and experience of the attending physician.

Compounds of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) and (Va) can form stable pharmaceutically acceptable salts with acids or bases, and in such cases it may be advantageous to administer the compound as a salt. Examples of acid addition salts include acetate, adipate, ascorbate, benzoate, benzenesulfonate, bicarbonate, bisulfate, butyrate, camphor, camphorsulfonate, choline, citrate, cyclohexylsulfamate, diethylenediamine, ethanesulfonate, fumarate, heptamhydone, glycolate hexanoate, hydrochloride, hydrobromide, hydroiodide, hydroxymaleate, lactate, malate, maleate, methanesulfonate, meglumine, 2-naphthalenesulfonate, nitrate, oxalate, pamoate, persulfate, phenylacetate, phosphate, diphosphate, picrate, pivalate, propionate,

- 16 037916 salicylate, stearate, succinate, sulfamate, sulfanilate, sulfate, tartrate, tosylate (p-toluenesulfonate), trifluoroacetate and undecanoate. Examples of basic salts include ammonium salts; alkali metal salts such as sodium, lithium and potassium salts; alkaline earth metal salts such as aluminum, calcium and magnesium salts; salts with organic bases such as salts of dicyclohexylamine and N-methyl-D-glucamine; and salts with amino acids such as arginine, lysine, ornithine, and so on. Also, basic nitrogen-containing groups can be quaternized with agents such as lower alkyl halides such as methyl, ethyl, propyl and butyl halides; dialkyl sulfates such as dimethyl, diethyl, dibutyl; diamyl sulfates; long chain halides such as decyl, lauryl, myristyl and stearyl halides; arylalkyl halides such as benzyl bromide, and others. Non-toxic, physiologically acceptable salts are preferred, although other salts may also find use, for example, in the isolation or purification of a product.

Salts can be prepared by conventional methods such as reacting the free base product with one or more equivalents of the corresponding acid in a solvent or medium in which the salt is insoluble, or in a solvent such as water, which is removed in vacuo or by lyophilization, or by exchanging the anions of the existing salt for another anion on a suitable ion exchange resin.

When a compound disclosed herein is described by a name or structure and has one or more chiral centers, and when the name or structure covers more than one stereoisomer, for example, the stereochemistry of one or more chiral centers is not indicated, the name or structure is intended to cover all such stereoisomers and mixtures thereof.

Synthesis of optically active forms can be carried out by standard organic chemistry methods well known in the art, for example, by synthesis from optically active starting materials or resolution of the racemic form. Racemates can be resolved into individual enantiomers using known methods (see, for example, Advanced Organic Chemistry: 3rd Edition: author J March, p104-107). A suitable method involves the formation of diastereomeric derivatives by reacting a racemic substance with a chiral excipient, followed by separation, for example chromatography, of the diastereomers and then elimination of the excipients. Likewise, the aforementioned activity can be assessed using standard laboratory techniques mentioned below.

Stereoisomers can be separated using conventional techniques such as chromatography or fractional crystallization. Enantiomers can be isolated by resolution of the racemate, for example, by fractional crystallization, separation, or HPLC. Diastereoisomers can be isolated by separation due to the different physical properties of the diastereoisomers, for example, by fractional crystallization, HPLC or flash chromatography. Alternatively, specific stereoisomers can be prepared by chiral synthesis from chiral starting materials under conditions that do not cause racemization or epimerization, or by derivatization with a chiral reagent.

When a particular stereoisomer is indicated, structurally or by name, it is preferably separated or substantially isolated from other stereoisomers of the same compound. In one aspect, a mixture comprising a particular stereoisomer of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) , may contain less than 30 wt.%, in particular less than 20 wt.% and in particular less than 10 wt.% other stereoisomers of the same compound. In another aspect, a mixture comprising a particular stereoisomer of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) , may contain less than 6 wt.%, in particular less than 3 wt.% and in particular less than 2 wt.% other stereoisomers of this compound. In another aspect, a mixture comprising a particular stereoisomer of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) , may contain less than 1 wt.%, in particular less than 0.5 wt.% and in particular less than 0.3 wt.%, in particular less than 0.1 wt.% other stereoisomers of this compound.

In one aspect, the terms infection and bacterial infection can refer to a gynecological infection. In yet another aspect, the terms infection and bacterial infection can refer to respiratory tract infection (IDI). In yet another aspect, the terms infection and bacterial infection can refer to a sexually transmitted disease. In yet another aspect, the terms infection and bacterial infection can refer to an uncomplicated urinary tract infection (UTI). In yet another aspect, the terms infection and bacterial infection can refer to a complicated urinary tract infection (UTI). In an additional aspect, the terms infection and bacterial infection may refer to an acute attack of chronic bronchitis (OPCB). In yet another additional aspect, the terms infection and bacterial infection can refer to acute otitis media of the middle ear. In one aspect, the terms infection and bacterial infection can refer to acute sinusitis. In yet another aspect, the terms infection and bacterial infection can refer to an infection caused by a drug-resistant bacteria. In yet another aspect, the terms infection and bacterial infection can refer to catheter-mediated sepsis. In yet another aspect, the terms infection and bacterial infection of mo

- 17 037916 gut refers to chancroid. In an additional aspect, the terms infection and bacterial infection can refer to chlamydia. In yet another additional aspect, the terms infection and bacterial infection can refer to community-acquired pneumonia (CAP). In yet another additional aspect, the terms infection and bacterial infection can refer to a complicated infection of the skin and skin structures. In one aspect, the terms infection and bacterial infection can refer to uncomplicated infection of the skin and dermal structures (ICS). In yet another aspect, the terms infection and bacterial infection can refer to endocarditis. In yet another aspect, the terms infection and bacterial infection can refer to febrile neutropenia. In yet another aspect, the terms infection and bacterial infection can refer to gonococcal cervicitis. In an additional aspect, the terms infection and bacterial infection can refer to gonococcal urethritis. In yet another additional aspect, the terms infection and bacterial infection can refer to nosocomial pneumonia (NP). In yet another aspect, the terms infection and bacterial infection can refer to osteomyelitis. In an additional aspect, the terms infection and bacterial infection can refer to sepsis. In yet another additional aspect, the terms infection and bacterial infection can refer to syphilis. In an additional aspect, the terms infection and bacterial infection can refer to intra-abdominal infection (IAI). In one aspect of the invention, the terms infection and bacterial infection may refer to an infection selected from the group consisting of complicated urinary tract infection, uncomplicated urinary tract infection, kidney infection, lower respiratory tract infection, nosocomial bacterial pneumonia, pneumonia, acute bacterial prostatitis, acute bacterial infection of the skin and soft tissues, sepsis, intra-abdominal infection and infection associated with diabetic foot.

In one embodiment, the terms infection and bacterial infection refer to an infection caused by gram-negative bacteria, also called gram-negative infection. In one aspect of this embodiment, the gram-negative infection is an infection that is resistant to one or more antibiotics. In one aspect of this embodiment, the gram-negative infection is a multi-drug resistant infection. In one aspect of this embodiment, the gram-negative infection is caused by one or more pathogens of Enterobacteriaceae spp. In one aspect of this embodiment, one or more pathogens of Enterobacteriaceae spp. include one or more of E. coli, K. pneumoniae, K. oxytoca, C. freundii, C. koseri, E. cloacae, P. mirabilis, M. morganii and / or S. marcescens. In one aspect of this embodiment, the one or more Enterobacteriaceae spp pathogens comprise one or more E. coli or K. pneumoniae pathogens. In yet another aspect of this embodiment, the gram-negative infection is caused by one or more biohazardous pathogens. In one aspect of this embodiment, one or more biohazardous pathogens are Burkholderia spp., Y. pestis and / or F. tularensis. In any of these aspects of an embodiment, one or more gram-negative pathogens may express one or more serine beta-lactamase enzymes. In one aspect of this embodiment, one or more serine beta-lactamase enzymes comprise one or more class A, class C and / or class D beta-lactamases.

All of the aforementioned infections can be caused by various bacteria that can potentially respond to treatment with the claimed agents, alone or in combination with penicillin binding protein inhibitors. One of the embodiments of the invention relates to a method of treating one or more of the above infections, comprising administering to a subject suffering from a bacterial infection, an effective amount of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa ), (IV), (IVa), (V) or (Va), or a pharmaceutically acceptable salt thereof in combination with an additional antibiotic. In one aspect of this embodiment, the additional antibiotic is a beta-lactam antibiotic. In one aspect of this embodiment, the additional antibiotic is a penicillin binding protein inhibitor.

In one aspect, there is provided the use of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or a pharmaceutically acceptable salt for the manufacture of a medicament for achieving an inhibitory effect on bacterial peptidoglycan, alone or in combination with a penicillin-binding protein inhibitor, in a warm-blooded animal such as a human.

In another aspect, there is provided the use of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or a pharmaceutically acceptable salt for the manufacture of a medicament for the treatment of a bacterial infection in a warm-blooded animal such as a human. In one aspect, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or a pharmaceutically acceptable salt thereof is administered in combination with an additional antibiotic such as a betalactam antibiotic. In one aspect of this embodiment, the additional antibiotic is a penicillin binding protein inhibitor. In one aspect of this embodiment, the additional antibiotic is a beta-lactam antibiotic. In one of

18 037916 aspects of this embodiment, the beta-lactam antibiotic is selected from cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibuten, cefditoren, pharopenem, ampoxycylidenem, cefditoren, ampopenem, ampoxycylidenem, cefditoren, ampopenem, tebicylidenem, cefditoren prodrugs. In one aspect of this embodiment, the betalactam antibiotic is cefpodoxime proxetil. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime axetil. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime or a prodrug thereof. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime or a prodrug thereof.

In another aspect, there is provided the use of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va), or a pharmaceutically acceptable salt for the manufacture of a medicinal product for the treatment of complicated urinary tract infection, uncomplicated urinary tract infection, kidney infection, lower respiratory tract infection, nosocomial bacterial pneumonia, pneumonia, acute bacterial prostatitis, acute bacterial infection of the skin and soft tissues, sepsis, intra-abdominal infection and infections associated with diabetic foot in a warm-blooded animal such as humans. One aspect of the invention relates to the use of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va), or a pharmaceutically acceptable salt for the manufacture of a medicament for the treatment of complicated urinary tract infections. In one aspect of the two preceding embodiments, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) is administered in combination with an additional antibiotic. In one aspect of this embodiment, the additional antibiotic is a penicillin binding protein inhibitor. In one aspect of this embodiment, the additional antibiotic is a beta-lactam antibiotic. In one aspect of this embodiment, the beta-lactam antibiotic is selected from cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibuten, cefditorin, carbenem, ceplipenem, ampoxicillin, and amoxicillin prodrugs. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime proxetil. In one aspect of this embodiment, the betalactam antibiotic is cefuroxime axetil. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime or a prodrug thereof. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime or a prodrug thereof.

In another aspect, there is provided a method of achieving an inhibitory effect on bacterial peptidoglycan, alone or in combination with an inhibitor of penicillin-binding protein, in a warm-blooded animal, such as a human, wherein said method comprises administering to said animal an effective amount of a compound of formulas (I), (Ia) , (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va), or a pharmaceutically acceptable salt thereof.

In an additional aspect, there is provided a method of treating a bacterial infection in a warm-blooded animal, such as a human, comprising administering to said animal an effective amount of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV ), (IVa), (V) or (Va), or a pharmaceutically acceptable salt thereof. In one aspect of this embodiment, the compound is as described for formulas (I), (II), (III), (IV), or (V), or a pharmaceutically acceptable salt thereof. In one aspect of this embodiment, for a compound of formulas (I), (II), (III), (IV), or (V), the compound is administered orally. In yet another aspect of this embodiment, a compound of formulas (I), (II), (III), (IV), or (V), or a pharmaceutically acceptable salt thereof, is administered in combination with an additional antibiotic. In one aspect of this embodiment, the additional antibiotic is a penicillin binding protein inhibitor. In one aspect, the additional antibiotic is a beta-lactam antibiotic. In one aspect of this embodiment, the beta-lactam antibiotic is selected from cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibuten, cefditorin, carbenem, ceplipenem, ampoxicillin, and amoxicillin prodrugs. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime proxetil. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime axetil. In one aspect of this embodiment, the betalactam antibiotic is cefpodoxime or a prodrug thereof. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime or a prodrug thereof.

In an additional embodiment, there is provided a method of treating a bacterial infection in a subject in need thereof, comprising administering to said subject an effective amount of a compound of formulas (Ia), (IIa), (IIIa), (IVa) or (Va) or a pharmaceutically acceptable salt thereof intravenously, intraperitoneally , intramuscularly or subcutaneously, preferably intravenously. As such, the compound of formulas (Ia), (IIa), (IIIa), (IVa) or (Va), or a pharmaceutically acceptable salt thereof, preimu

- 19 037916 are commonly used when the patient cannot take medications orally, for example, in a hospital setting (for example, in the intensive care unit, in the emergency department, in the cardiology department, etc.), in the conditions of an emergency room and conditions nursing home. In one aspect of this embodiment, a compound of formulas (Ia), (IIa), (IIIa), (IVa), or (Va), or a pharmaceutically acceptable salt thereof, is administered intravenously (iv). In one aspect of this embodiment, a compound of formulas (Ia), (IIa), (IIIa), (IVa), or (Va), or a pharmaceutically acceptable salt thereof, is administered until the patient can take medications orally, for example, in the duration of the stay in the hospital or emergency room, until the subject can be discharged from the hospital or emergency room, or until the patient has improved enough to be able to take medications by mouth, for example, until until the patient can take medications orally in a nursing home. In one aspect of this embodiment, a compound of formulas (Ia), (IIa), (IIIa), (IVa), or (Va), or a pharmaceutically acceptable salt thereof, is administered with a beta-lactam antibiotic. In one aspect of this embodiment, the beta-lactam antibiotic is selected from cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibuten, cefditorin, carbenem, ceplipenem, ampoxicillin, and amoxicillin prodrugs. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime or a prodrug thereof. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime or a prodrug thereof. In yet another aspect of this embodiment, the method further comprises administering to the subject an effective amount of a compound of formula (I), (II), (III), (IV), or (V), or a pharmaceutically acceptable salt thereof, on an outpatient basis, after being discharged from a hospital or hospital. emergency care, or when the patient's condition has improved enough to be able to take medications by mouth, for example, in a nursing home. In one aspect of this embodiment, a compound of formula (I), (II), (III), (IV), or (V), or a pharmaceutically acceptable salt thereof, is orally administered on an outpatient basis. In one aspect of this embodiment, the compound of formula (I), (II), (III), (IV), or (V), or a pharmaceutically acceptable salt thereof, is administered in combination with the same beta-lactam antibiotic with which the compound of formulas ( Ia), (IIa), (IIIa), (IVa) or (Va) in an inpatient, emergency room, or nursing home setting. In one aspect of this embodiment, after administration of a compound of formulas (Ia), (IIa), (IIIa), (IVa), or (Va), a compound of formula (I), (II), (III), (IV) is orally administered or (v) or a pharmaceutically acceptable salt thereof, as soon as the patient is able to take medications orally, for example, after the patient has been discharged from a hospital or emergency room and is on an outpatient basis, or as soon as the patient's condition in a nursing home has improved enough to be able to take medications orally. Preferably, the transition from intravenous, intraperitoneal, intramuscular or subcutaneous (compound of formulas (Ia), (IIa), (IIIa), (IVa) or (Va)) to oral administration (compound of formulas (I), (II), (III) , (Iv) or (v)) occurs without any interruption in the patient's treatment.

In yet another additional aspect, a method is provided for the treatment of complicated urinary tract infection, uncomplicated urinary tract infection, kidney infection, lower respiratory tract infection, nosocomial bacterial pneumonia (NBP), pneumonia, acute bacterial prostatitis, acute bacterial infection of the skin and soft tissues, sepsis, intra-abdominal infections and infections associated with a diabetic foot in a warm-blooded animal, such as a human, wherein said method comprises administering to said animal an effective amount of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa ), (IV), (IVa), (V) or (Va), or a pharmaceutically acceptable salt thereof. In yet another additional aspect, there is provided a method of treating complicated urinary tract infections in a warm-blooded animal, such as a human, comprising administering to said animal an effective amount of a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa) ), (IV), (IVa), (V) or (Va), or a pharmaceutically acceptable salt thereof. In one aspect of any of the preceding embodiments, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) or a pharmaceutically acceptable salt thereof is administered in combination with an additional antibiotic. In one aspect of this embodiment, the additional antibiotic is a penicillin binding protein inhibitor. In one aspect, the additional antibiotic is a beta-lactam antibiotic. In one aspect of this embodiment, the beta-lactam antibiotic is selected from cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibuten, cefditorin, carbenem, ceplipenem, ampoxicillin, and amoxicillin prodrugs. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime proxetil. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime axetil. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime or a prodrug thereof. In one aspect of this embodiment,

- 20 037916 The beta-lactam antibiotic is cefuroxime or its prodrug.

In yet another additional aspect, there is provided a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or a pharmaceutically acceptable salt for use to achieve an inhibitory effect on bacterial peptidoglycan, alone or in combination with a penicillin-binding protein inhibitor, in a warm-blooded animal such as a human. In one aspect, there is provided a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or a pharmaceutically acceptable salt for use in the treatment of gram-negative bacterial infections, alone or in combination with a beta-lactam antibiotic. In one aspect of this embodiment, the beta-lactam antibiotic is selected from cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibuten, cefditorin, carbenem, ceplipenem, ampoxicillin, and amoxicillin prodrugs. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime proxetil. In one aspect of this embodiment, the betalactam antibiotic is cefuroxime axetil. In one aspect of this embodiment, the beta-lactam antibiotic is cefpodoxime or a prodrug thereof. In one aspect of this embodiment, the beta-lactam antibiotic is cefuroxime or a prodrug thereof.

In one aspect, the invention provides a method of inhibiting one or more beta-lactamase enzymes, comprising administering a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa) ), (V) or (Va) or a pharmaceutically acceptable salt thereof to an animal in need thereof. In an additional aspect, one or more beta-lactamase enzymes are beta-lactamase serine enzymes. In a further aspect, one or more beta-lactamase enzymes are selected from the group consisting of class A, class C, and class D. In a further aspect, one or more beta-lactamase enzymes are a class A. In a further aspect, one or more beta-lactamase enzymes are is a class C enzyme. In a further aspect, one or more beta-lactamase enzymes are a class D enzyme. In a further aspect, one or more beta-lactamase enzymes are a class D enzyme and one or more class A and C enzymes. In a further aspect, one or more beta-lactamase enzymes are enzymes of all three classes A, C and D.

The beta-lactamase inhibitors of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) can be administered in combination with any beta-lactam antibiotic belonging to, but not limited to, the classes of clavams, carbapenems, monobactams, penems, penicillins and / or cephalosporins, or any other compound susceptible to serine beta-lactamases. In one aspect of the invention, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) is combined with one or more of penicillin, methicillin, oxacillin, nafcillin, cloxacillin, dicloxacillin, flucloxacillin, temotsillina, amoxicillin, ampicillin, koamoksiklava, azlocillin, carbenicillin, ticarcillin, mezlocillin, piperacillin, cephalexin, cephalothin, SKHA101, cefazolin, cefaclor, cefuroxime, tsefamandola, cefotetan, cefoxitin, ceftriaxone, cefotaxime, cefpodoxime, cefixime, ceftazidime, ceftobiprol medocaryl, cefepime, cefpirome, ceftaroline, imipenem, meropenem, ertapenem, faropenem, Sulopenem 601, BESTAMETAL PREZAL36 -1, tomopenem, tebipenem, aztreonam, tigemonam, nocardicin A, or tabtoxinin beta-lactam. In one aspect of the invention, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) is combined with cefpodoxime, cefuroxime, tigemonam, cefixime, or faropenem. In one aspect of the invention, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) is combined with an antibacterial compound from the group consisting of penicillin V, cloxacillin, dicloxacillin, nafcillin, oxacillin, amoxicillin, ampicillin, bacampicillin, amoxicillin-clavulanate, carbenicillin, cefadroxil, cefalexin, cefradin, cefacrozimar, cephorazylar , or a prodrug or salt thereof. In one aspect of the invention, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) is combined with an antibacterial compound from the group consisting of cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibutene, cefditoren, faropenem, tebipenem, amoxicillin, carbenicillin, and ceficidestv. In one aspect of the invention, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) is combined with cefpodoxime, or its prodrug such as cefpodoxime proxetil. In one aspect of the invention, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) is combined with cefuroxime, or its prodrug such as cefuroxime axetil.

In another aspect of the invention, the compound of formulas (I), (II), (III) or (IV) is administered in combination with a beta-lactam antibiotic and an additional antibiotic and / or an additional inhibitor of beta-lactamases. In one aspect of the invention, the additional antibiotic is selected from one of the classes of aminoglycosides, spectinomycins, macrolides, ketolides, streptogramins, oxazolidinones,

- 21 037916 tetracyclines, fluoroquinolones, quinolones, coumarin antibiotics, glycopeptides, lipoglycopeptides, nitroimidazoles, ansamycins, fenikols, mupirocin, fosfomycin, tobramycin, linezolid, daptomycin, ASIALT Ed: A. Bryskier (2005)).

In one aspect of the invention, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) is administered in combination with a beta-lactam antibiotic and a second agent designed to overcome beta-lactam resistance. In one aspect of the present invention, the second agent for overcoming beta-lactam resistance may be a metallo-beta-lactamase (MBL) inhibitor, also known as a class B inhibitor.

In one aspect, there is provided a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or pharmaceutically an acceptable salt for use in treating a bacterial infection in a warm-blooded animal, such as a human.

In another aspect, there is provided a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va), or a pharmaceutically acceptable salt for use in the treatment of complicated urinary tract infection, uncomplicated urinary tract infection, kidney infection, lower respiratory tract infection, nosocomial bacterial pneumonia, pneumonia, acute bacterial prostatitis, acute bacterial infection of the skin and soft tissues, sepsis, intra-abdominal infection and infections associated with a diabetic foot, in a warm-blooded animal such as a human. In another aspect, there is provided a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or pharmaceutically an acceptable salt for use in the treatment of complicated urinary tract infections in a warm-blooded animal such as a human.

In another aspect, there is provided a pharmaceutical composition comprising a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier, diluent or excipient. In one aspect of this embodiment, the pharmaceutical composition further comprises a betalactam antibiotic. In one aspect of this embodiment, the beta-lactam antibiotic is selected from cefpodoxime, cefuroxime, tigemonam, loracarbef, cefixime, cephalexin, cefadroxil, cefetamet, cefprozil, ceftibuten, cefditorin, carbenem, ceplipenem, ampoxicillin, and amoxicillin prodrugs.

The compositions according to the invention can be presented in a form suitable for oral administration (for example, tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use ( for example, in the form of creams, ointments, gels or aqueous or oily solutions or suspensions), for administration by inhalation (for example, as a finely divided powder or liquid aerosol), for administration by insufflation (for example, as a finely divided powder) or for parenteral administration (for example, as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular or intramuscular administration, or as a suppository for rectal administration). In one aspect of the invention, a compound of formulas (Ia), (IIa), (IIIa), (IVa) or (Va), or a pharmaceutically acceptable salt thereof, is administered intravenously. In yet another aspect of the invention, a compound of formulas (Ia), (IIa), (IIIa), (IVa) or (Va), or a pharmaceutically acceptable salt thereof, is administered intravenously in combination with one or more other antibacterial agents. In one aspect of the invention, a compound of formulas (I), (II), (III), (IV) or (V), or a pharmaceutically acceptable salt thereof, is administered orally. In yet another aspect of the invention, a compound of formulas (I), (II), (III), (IV) or (V), or a pharmaceutically acceptable salt thereof, is administered orally in combination with one or more other antibacterial agents. In one aspect of any of these embodiments, a compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va) or a pharmaceutically acceptable salt thereof is administered concurrently with one or more other antibacterial drugs. In yet another aspect of this embodiment, the compound of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V), or (Va), or the pharmaceutically acceptable salt is administered sequentially with one or more other antibacterial agents such as a beta-lactam antibiotic.

One of the embodiments of the invention relates to a method of treating a bacterial infection in a person in need thereof, comprising administering to said person an effective amount of a compound of one of formulas (Ia), (IIa), (IIIa), (IVa) or (Va) intravenously in combination with one or more additional antibacterial drugs on an outpatient basis, in an emergency room, or in a nursing home setting, followed by the administration to said person of an effective amount of a compound of one of formulas (I), (II), (III), (IV) or (V) orally in combinations with one or more additional antibacterial drugs outside, for example, an outpatient setting, an emergency room setting, or a nursing home setting, as soon as the patient is able to take oral drugs again.

One embodiment of the invention relates to a method of treating a bacterial infection in a person in need thereof, comprising orally administering to said person an effective amount of a compound of one of formulas (I), (II), (III), (IV) or (V ) in combination with one or more additional antibacterial drugs as a stepwise therapy with the transition to the oral route of administration after the administration to the specified person of an effective amount of one or more antibacterial drugs administered intravenously, intraperitoneally, intramuscularly or subcutaneously, for example, compounds of formulas (Ia), ( IIa), (IIIa), (IVa) or (Va) or a pharmaceutically acceptable salt thereof. The nature, dose and duration of antibiotic therapy and the timing of the transition from the intravenous, intraperitoneal, intramuscular or subcutaneous route of administration to an oral drug are usually selected by the physician and may depend on the patient's health status, his or her ability to receive oral treatment and the type of infections from which the specified Human. The patient can be switched from intravenous, intraperitoneal, intramuscular, or subcutaneous to oral treatment when the patient no longer shows symptoms, does not have a fever, or has a slightly fever (e.g., below 100.5 ° F (approximately 38 ° C) , 100 ° F (approximately 37.8 ° C), 99.5 ° F (approximately 37.5 ° C), etc.) has been removed from the ventilator or no longer requires intravenous fluids.

The compositions of the invention can be prepared by conventional methods using conventional pharmaceutical excipients well known in the art. Suitable pharmaceutically acceptable tabletting aids include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate; granulating and disintegrating agents such as corn starch or alginic acid; binding agents such as starch; lubricants such as magnesium stearate, stearic acid or talc; preservatives such as ethyl or propyl n-hydroxybenzoate; and antioxidants such as ascorbic acid. Tablet formulations may be uncoated or coated to alter their disintegration and subsequent absorption of the active ingredient in the gastrointestinal tract, or to improve their stability and / or appearance, in any case using conventional coating agents and techniques well known in the art.

The amount of active ingredient combined with one or more excipients to form a unit dosage form will vary as needed depending on the recipient to be treated and the particular route of administration. For example, a formulation intended for oral administration to humans will typically contain, for example, the active agent mixed with a suitable and convenient amount of excipients, which may range from about 5 to about 98% by weight based on the total composition. Dosage units usually contain from about 100 to about 4000 mg of the active ingredient. For oral administration, for example, a compound of formulas (I), (II), (III), (IV) or (V) or pharmaceutically acceptable salts thereof, 0.1 to 10 g-equiv of active ingredient per day is suitable; and for intravenous administration, for example, a compound of formulas (Ia), (IIa), (IIIa), (IVa) or (Va) or their pharmaceutically acceptable salts, from 0.5 to 8 g-equiv of the active ingredient per day is suitable.

In addition to the compounds of the present invention, the pharmaceutical composition of the present invention may also contain or be co-administered with (simultaneously, sequentially or separately) one or more known drugs selected from other clinically useful classes of antibacterial drugs (e.g. macrolides, quinolones, betalactams or aminoglycosides) and / or other anti-infective drugs (for example, antifungal triazole or amphotericin). These may include carbapenems such as meropenem or imipenem to enhance therapeutic efficacy. The compounds of the present invention may also contain or be co-administered with bactericidal / permeation-enhancing (BPI) protein products or efflux pump inhibitors to improve activity against gram-negative bacteria and antimicrobial resistant bacteria.

As indicated above, the size of the dose required for the therapeutic or prophylactic treatment of a particular pathological condition will vary as needed depending on the recipient to be treated, the route of administration, and the severity of the disease to be treated. Accordingly, the optimal dosage can be determined by the practitioner treating any particular patient.

Compounds of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) can be prepared in several ways. The processes shown below illustrate a method for synthesizing compounds of formula (I) (wherein R ¹ , R ² and R ³ , unless otherwise indicated, are as defined above). The reactions are carried out in solvents appropriate to the reagents and materials used and are suitable for the transformations to be carried out. In addition, when describing the synthesis methods given below, it is understood that all the proposed reaction conditions, including the choice of solvents, the atmosphere in which the reaction is carried out, the reaction temperature, the duration of the experiment and the processing methods, are selected as conditions standard for this reaction. that should be obvious to a person skilled in the art. One skilled in the art of organic synthesis understands that functional groups present in various parts of the molecule must be compatible with the disclosed reagents and reactions. Such restrictions on substituents compatible with the reaction conditions will be apparent to one skilled in the art, in which case alternative methods must be used. Schemes and processes should not cover an exhaustive list of methods for preparing compounds of formulas (I), (Ia), (II), (IIa), (III), (IIIa), (IV), (IVa), (V) or (Va) ); in contrast, additional techniques known to the skilled chemist can also be used to synthesize the compounds. The claims are not limited to the structures shown in the diagrams and processes.

It should also be understood that in some of the reactions shown in the schemes and in the processes mentioned herein, it may be necessary / desirable to protect any sensitive groups in the compounds. Those skilled in the art know when protection is necessary or desirable, as well as suitable methods for such protection. Conventional protecting groups can be used in accordance with standard practice (for example, see T. W. Greene, Protective Groups in Organic Synthesis, published by John Wiley and Sons, (1991)) and as described hereinabove.

The skilled chemist will be able to use and adapt the information contained in the above sources, as well as the information to which they are referenced and contained in the accompanying examples, as well as in the examples and schemes provided in this document, to obtain the necessary starting materials and products.

The required starting materials for methods such as those described herein, if they are not commercially available, can be prepared using methods selected from standard organic chemistry techniques, analogous to the synthesis of known, structurally similar compounds, or analogous techniques described. method or methods described in the examples.

It is noted that many of the starting materials for synthetic methods as described herein are commercially available and / or widely described in the scientific literature, or can be obtained from commercially available compounds using adaptation of the processes described in the scientific literature. Next, the reader is referred to Advanced Organic Chemistry source, 5 ^th Edition, for the author. Jerry March and Michael Smith published by John Wiley & Sons (2001) for general guidance on reaction conditions and reagents.

General techniques and schemes

In one aspect, compounds of formulas (I) and (Ia), or pharmaceutically acceptable salts thereof, can be prepared by the method described in Scheme 1. From Weinreb amide, the introduction of substituents at the R ³ position of formulas (I) and (Ia) can be carried out using Grignard reactions. The ester moieties can be introduced by palladium-catalyzed deallylation followed by alkylation with bromoacetates. Hydrolysis of esters produces acids. Alternatively, other R ¹ groups can be obtained by modifying the primary alcohol.

Scheme 1

[C ₃ H ₅ PdCI] ₂ ligand (R, R), O phthalimide,

Na ₂ CQ ₃ DXM,

TBSCI, TBSO ^ V%

ОМе г, TBSO ^ VA hydrazine, Br yjМеОЦббС TBSO ^ VA о - Boc ^N >

NH ₂ Boc2O, DMFN '° primary amine। Weinreb's amide

R ³ ^^^ MdBg THF, О С

catalyst ₃

Howeida-Grubbs TBSO ^/ ''' ^, r ^z ' ^^ generation, II CeCI ₃ NaBH ₄ toluene, 65 C Boc ^x ^^ ОМеОН, OC

AIIONH-Ns, PPh ₃ DIAD, toluene, room temp

СгО ₃ Н ₅ Ю ₆

ZnBr ₂

primary alcohol

PhSH

triphosgene

1. Pd [PPh ₃ ] ₄ MeOH

1,3-dimethylbarbituric acid base, DMF

O

UOH,

THF, H ₂ O

Alternative methods for the synthesis of compounds with substitution at R ³ are shown in Scheme 2, where the key steps are the Diels-Alder reaction and the nitrosoene reaction.

- 24 037916

Scheme 2

T (3 4 eq)

TMSOTf

Diels-Alder

O

EY

LiOH R ³ THF H ₂ O

CPI NH ₄ OAc _r

BocNHOH O ₂ О or air dSC r3

---------- ► H ₂ N 'g' ck 'nitroso-enic boc' ^N ^ ^x ^ ⁴ n '° ^H

Vos

TBSCI πμπ ^η 2 ^ν ''

Vos'

Vos

Acetate and ester moieties can be introduced in one or two steps according to Scheme 3.

Scheme 3

In yet another aspect, compounds of formulas (I) and (Ia), or pharmaceutically acceptable salts thereof, can be prepared by the method described in Scheme 4, wherein substitution at R ² can be introduced by Michael addition to the enone 1 followed by oxidation to give the enone 2, after which the synthesis is similar to that described in Scheme 1.

Scheme 4

Alternatively, amide R ¹ can be introduced after cyclization of urea according to scheme

five.

Scheme 5

Compounds with substitution at R ² can also be synthesized from Garner's aldehyde as shown in Scheme 6. The path from the primary amine to compounds of formulas (I) or (Ia) is similar to that of Scheme 1.

Scheme 6 oL //, o ^ \ / ° L // _D οΛ # J1 JI

-J + R-MgBr-L? \ DessMartin ^ Щ p “Wittita j 4 1) TsOH TBSO '^^' R ² ZnBr ₂ TBSO '^ V ^ R ² N ------- ► R ² ---------- ► / NR ² R —------- ► ιΓιιι ----- * r „,

Boc Boc Boc Boc 2) Boc ₂ O _B oc ' ^NH NH ₂

o) I Dobl

In addition, compounds with substitution at R ² can be synthesized according to the following scheme 7, where the key step is the nitroso-ene reaction. The amide can also be introduced at an earlier stage of the synthesis from the carboxylic acid and stored until the end of the synthesis.

- 25 037916

Scheme 7

Alkyl bromoacetates for the introduction of carboxylic acids and ester fragments can be obtained by transesterification according to Scheme 8.

Scheme 8

Chiral bromofluoroacetates for the introduction of R- and S-fluorocarboxylic acid and ester fragments can be obtained by recrystallization of bromofluoroacetic acid with chiral phenylethylamine followed by esterification according to Scheme 9.

Scheme 9

In any of the aforementioned pharmaceutical compositions, processes, methods, uses, medicaments and manufacturing features of the present invention, any alternative embodiments of the compounds of the invention described herein are also applicable. For example, additional details and method of carrying out the nitroso-ene reaction on various substrates are as described below in the examples section. These details and methods include, for example, a first method for preparing a compound of formula VI.

R ^{2 ri} ^. / K / ^r3

X, / PG 'PG N

I

OH (VI);

or its salt, where

R ¹ is -C (O) NR ⁷ R ⁸ , -C (O) OR ⁷ , -CH2OR7, -CN, phenyl, 5-7 membered heteroaryl, -C (O) NR'NR'C (O) R ⁹ , -C (O) NR'OR ¹⁰ or a C1-C6 alkyl group, where the alkyl group is substituted by 1-3 groups consisting of halogen, C1-C3 alkoxy, -OH, -CN, -NR ⁷ R ⁸ , -NR7COR ⁹ , 5-7 membered heteroaryl and 5-7 membered heterocyclyl, and wherein the phenyl and heteroaryl represented by R ¹ are optionally and independently substituted with 1-3 groups selected from halogen, -OH, C1-C ₃ - alkoxy, —CN, —NR7R ⁸ and —CONR7R ⁸ ;

each of R ² and R ^{3 is} independently selected from hydrogen, halogen, C ₁ -C ₃ -alkyl and C ₃ -C ₆ -cycloalkyl, with the proviso that at least one of R ² and R ^{3 is} not hydrogen;

each R ⁷ and R ^{8 is} independently hydrogen, C1-C3 alkyl, C1-C3 alkoxy, phenyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, where alkyl, alkoxy, phenyl, cycloalkyl, heterocyclyl or heteroaryl represented by R ⁷ or R ⁸ is optionally and independently substituted with 16 groups selected from 5-6 membered heterocyclyl optionally substituted with one or two —F, carboxyl or —CO (OC1-6alkyl), 5- 6-membered heteroaryl, -CN, -OH, C ₁ -C ₃ -alkyl optionally substituted with -NH2 or -OH, C ₁ -C ₃ -halogenoalkyl, C ₁ -C ₃ -haloalkoxy, C ₁ -C ₃ -alkoxy -NHCO (C1-Cs-alkyl), -NHCO (C1-Cs-alkoxy), -S (O) 2NR'R, -NHS (O) 2NR'R, -NHS (O) 2 (C1-Cs-αalkyl ), -NR'R and -C (O) NR'R;

each R ⁹ is C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ haloalkoxy, or C ₁ -C ₆ alkoxy;

each R 'and R is independently hydrogen, methyl, ethyl or propyl; or R 'and R inter

- 26 037916 STE with the nitrogen atom to which they are attached form a 5-6-membered heterocyclyl; and each of PG and PG 'independently represents an amino protecting group; where the method comprises reacting a compound of formula XI

or its salts with PG'NHOH in the presence of an oxidizing agent to form a compound of formula VI; Also provided is a second method for preparing a compound of formula VI

or its salt, where

R ¹ is -C (O) NR ⁷ R ⁸ , -C (O) OR ⁷ , -CH2OR7, -CN, phenyl, 5-7 membered heteroaryl, -C (O) NR'NR'C (O) R ⁹ , -C (O) NR'OR ¹⁰ or C1-C6-alkyl group, where the alkyl group is substituted by 1-3 groups consisting of halogen, C1-C3-alkoxy, -OH, -CN, -NR ⁷ R ⁸ , -NR7COR ⁹ , 5-7 membered heteroaryl and 5-7 membered heterocyclyl, and wherein the phenyl and heteroaryl represented by R ¹ are optionally and independently substituted with 1-3 groups selected from halogen, -OH, C1-C3-alkoxy , -CN, -NR ⁷ R ⁸ and -CONR7R ⁸ ;

each of R ² and R ^{3 is} independently selected from hydrogen, halogen, C ₁ -C ₃ -alkyl and C _{3 -C 6} -cycloalkyl, with the proviso that at least one of R ² and R ^{3 is} not hydrogen;

each R ⁷ and R ^{8 is} independently hydrogen, C1-C3 alkyl, C1-C3 alkoxy, phenyl, C3-C6 cycloalkyl, 4-7 membered heterocyclyl, or 5-6 membered heteroaryl, where alkyl, alkoxy, phenyl, cycloalkyl, heterocyclyl, or heteroaryl represented by R ⁷ or R ⁸ is optionally and independently substituted with 16 groups selected from 5-6 membered heterocyclyl optionally substituted with one or two —F, carboxyl or —CO (OC1-6 alkyl), 5- 6-membered heteroaryl, -CN, -OH, _C1-C3 alkyl optionally substituted with-NH ₂ or -OH, C 1 C ₃ -haloalkyl, C 1 C ₃ -galogenalkoksi, _C1-C3 alkoxy, -NHCO ( C1-Cs-alkyl), -NHCO (C1-Cs-alkoxy), -S (O) 2NR'R, -NHS (O) 2NR'R, -NHS (O) 2 (C1-Cs-alkyl), - NR'R and -C (O) NR'R;

each R ⁹ is _C1-C6 alkyl, _C1-C6 haloalkyl, _C1-C6 -galogenalkoksi or C1-C6alkoxy;

each R 'and R independently represents hydrogen, methyl, ethyl or propyl; or R 'and R together with the nitrogen atom to which they are attached form a 5-6 membered heterocyclyl; and each of PG and PG 'independently represents an amino protecting group;

where the method comprises reacting a compound of formula XI

or its salts with PG'N = O to form a compound of formula VI.

In a first aspect, the compound of formula XI in the first or second method has the formula

or its salt.

In a second aspect, the compound of formula VI in the first or second method has formula VII

or its salt.

In a third aspect, R ² in the first or second method or formula VII is C1-C3 alkyl, the remaining features being as described in the first or second method and the first or second aspect. Alternatively, R ² in the first or second method or formula VII is methyl, the remaining features being as described in the first or second method and the first or second aspect.

In a fourth aspect, the compound of formula VI in the first or second method has formula VIII

- 27 037916 ^r V /% / ^r3 , Ν. / L .PG '

PG N

I

OH (USH);

or a salt thereof, the remaining features being as described in the first or second method and the first or second aspect.

In a fifth aspect, R ³ in the first or second method or formula VII is C ₁ -C ₃ -alkyl, the remaining features being as described in the first or second method and the first or second aspect. Alternatively, R ³ in the first or second method or formula VIII is methyl, the remaining features being as described in the first or second method and the first or second aspect.

In a sixth aspect, R ¹ in the first or second method is selected from oxadiazole, -C (O) NHNHC (O) (C1-C3 alkyl), -CH2NH2, -CH2NHCO (C1-Cs-alkoxy), -CH2NHCO (C1-Cs-alkyl ) or -CH2NHCO (C1-C3-haloalkyl), where oxadiazole R ^{1 is} optionally substituted with -OH, C1-C ₃ alkoxy, -NR ⁷ R ⁸ or -CONR7R ⁸ ; and wherein the remaining features are as described in the first or second method and the first, second, third, fourth or fifth aspect.

In a seventh aspect, R ¹ in the first or second method is selected from —CH2NH2,

and wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth or sixth aspect.

In an eighth aspect, R ¹ in the first or second method is: -CN,

where R ¹¹ represents hydrogen or -C (O) NH ₂ ; and wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth or seventh aspect.

In a ninth aspect, R ¹ in the first or second method is —C (O) NR ⁷ R ⁸ , —C (O) OR ^7, or —CN; and wherein the remaining features are as described in the first or second method and in the first, second, third, fourth, fifth, sixth, seventh or eighth aspect. Alternatively, R ¹ in the first or second method is —C (O) NH2, —C (O) OH, —CN, or —C (O) OC1-C6 alkyl; while the remaining features are the same as described in the first or second method and in the first, second, third, fourth, fifth, sixth, seventh or eighth method. In yet another alternative, R ¹ in the first or second process is —CN or —C (O) NH2; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh or eighth aspect. In yet another alternative, R ¹ in the first or second method is —CN; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh or eighth aspect. In yet another alternative embodiment, R ¹ in the first or second method is —C (O) NR ⁷ R ⁸ ; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh or eighth aspect.

In a tenth aspect, R ⁷ and R ⁸ in the first or second method are both hydrogen; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth or ninth aspect. Alternatively, R ⁷ in the first or second method is hydrogen and R ⁸ is: 1) phenyl optionally substituted with C1-C3 alkyl or C1-C3 alkyl NH2, 2) C1-C3 alkyl or 3 ) C1-C3-alkoxy, where each alkyl or alkoxy of R ^{8 is} optionally and independently substituted with C3-C ₆ -cycloalkyl, -CN, -OH, -NH2, -SO2NH2, -NHSO2NH2, -C (O) NH2, - NHC (O) (C1 -C3 alkyl), pyrazinyl, oxytanyl, oxazolyl or pyrrolidinyl optionally substituted with one or more carboxyl, fluorine or —C (O) O (C ₁ -C ₆ alkyl); while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth or ninth aspect. In yet another alternative embodiment, R ⁷ in the first or second method is hydrogen, and R ^{8 is} selected from the group consisting of

- 28 037916

-CH2CN and -CH2OH;

while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth or ninth aspect.

In an eleventh aspect, the compound of formula VI in the first or second method has formula IX

I he (IX);

or its salt.

In a twelfth aspect, the compound of formula VI in the first or second method has the formula X o

I he (X);

or its salt.

In a thirteenth aspect, each of PG and PG 'in the first or second method together with the amine nitrogen atom which they protect independently forms a carbamate, amide or N-benzyl or N-aryl; wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh or twelfth aspect. Alternatively, each of PG and PG 'in the first or second method is independently selected from tert-butyloxycarbonyl (Boc), carboxybenzyl (Cbz), fluorenylmethyloxycarbonyl (Fmoc), 2,2,2trichloroethoxycarbonyl (Troc), CF3CO, acetyl (Ac) , paratoluenesulfonamide (Ts) and methylsulfonyl (Ms); while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh or twelfth aspect. In yet another alternative, each of PG and PG 'in the first or second method is the same; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh or twelfth aspect. In yet another alternative, each of PG and PG 'in the first or second method is tert-butoxycarbonyl; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh or twelfth aspect.

In a fourteenth aspect, the first method further comprises reacting a compound of Formula XI with PG'NHOH in the presence of a metal catalyst; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth or thirteenth aspect. Alternatively, the metal catalyst is selected from CuCl, CuBr, CuI, CuCN, CuSCN, CuBrMe ₂ S, Cu (OAc) _2, and CuOTf; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth or thirteenth aspect. In yet another alternative, the metal catalyst comprises a copper salt; while the remaining features are the same as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth,

- 29 037916 ninth, tenth, eleventh, twelfth or thirteenth aspect. In yet another alternative, the metal catalyst comprises a copper halide salt; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth or thirteenth aspect. In yet another alternative, the metal catalyst is CuCl or CuBrMe2S; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth or thirteenth aspect.

In a fifteenth aspect, the first method further comprises reacting a compound of Formula XI with PG'NHOH in the presence of an amine additive; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspect. In one aspect, the amine additive is pyridine, (1R, 2R) -cyclohexane-1,2-diamine, N, N'dimethylethane-1,2-diamine, 2,6-di-tert-butyl-4-methylpyridine, 1,10-phenanthroline, trans-cyclohexane-1,2-diamine, N1- (2- (diethylamino) ethyl) -N2, N2-diethylethane-1,2-diamine, cis-cyclohexane-1,2-diamine or N1, N1 , N2, N2-tetramethylethane-1,2-diamine; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspect. Alternatively, the amine is selected from pyridine, 2,6-lutidine, 4-dimethylaminopyridine, picoline, 1,8 diazabicyclo [5.4.0] undec-7-ene, and N, N-diisonropylethylamine; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspect. In yet another alternative, the amine is pyridine; the remaining features are as described in the first or second method and in the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth or fourteenth aspect.

In a sixteenth aspect, the oxidizing agent in the first or second method is O ₂ , air, FeCl ₃ , MnO ₂ , meta-chloroperoxybenzoic acid (mCPBA), NaIO ₄ , 2-iodoxybenzoic acid (IBX), (2,2,6,6 tetramethylpiperidine-1 -yl) oxyl (TEMPO), benzoyl peroxide (BPO), HIO3, urea-H ₂ O ₂ , I ₂ , N-chlorosuccinimide (NCS), Dess-Martin periodinane (DMP), H ₂ O ₂ or N-methylmorpholine N-oxide ( NMMO); wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth or fifteenth aspect. Alternatively, the oxidizing agent is urea-H ₂ O ₂ , H ₂ O _2, or O ₂ ; wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth or fifteenth aspect.

In a seventeenth aspect, the reaction in the first or second method is carried out in a polar solvent; wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth or sixteenth aspect. Alternatively, the reaction is carried out in DCM, THF, MTBE, EtOAc, iPrOAc, MeCN, H ₂ O, MeOH, EtOH, i-PrOH, t-BuOH, nBuOH, 2-methyl-2-butanol, DMF, DMSO, ethylene glycol , polyethylene glycol, sulfolane, sulfolane / H ₂ O mixtures, DMF / H 2 O, NMP / H 2 O, DCM / H 2 O, MeOH / H ₂ O, EtOH / H ₂ O, iPrOH / H ₂ O or n-BuOH / H ₂ O; wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth or sixteenth aspect. In yet another alternative, the reaction is carried out in methylene chloride or sulfolane; wherein the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth or sixteenth aspect.

In an eighteenth aspect, the reaction in the first or second method further comprises adding water; while the remaining features are as described in the first or second method and the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, fifteenth, sixteenth or seventeenth aspect.

Examples of

The invention will hereinafter be further described with reference to the following illustrative examples, in which, unless otherwise indicated:

(i) temperature is indicated in degrees Celsius (° C); operations are carried out at room temperature or ambient temperature, that is, in the range of 18-25 ° C;

(ii) organic solutions were dried over anhydrous magnesium sulfate;

the organic solvent was evaporated using a rotary evaporator

- 30 037916 at reduced pressure (4.5-30 mm Hg) and at a bath temperature of up to 60 ° C;

(iii) chromatography means flash chromatography on silica gel; thin layer chromatography (TLC) was performed on silica gel plates;

(iv) usually the progress of the reaction was monitored by TLC or liquid chromatography / mass spectroscopy (LC / MS), and the reaction time is given by way of example only;

(v) the end products have satisfactory proton nuclear magnetic resonance (NMR) spectra and / or mass spectral data;

(vi) the outputs are for example only and do not necessarily represent outputs that can be obtained by diligently evolving the process; the preparation was repeated if more starting materials were required;

(vii) NMR data, if indicated, are presented as delta values for the main protons used for identification, given in parts per million (ppm) relative to tetramethylsilane (TMS) as an internal standard, determined at 300 MHz in DMSO- d ₆ , unless otherwise indicated;

(viii) chemical symbols have their usual meanings; (ix) the ratio of solvents is indicated in units of volume: volume (v / v); (x) ISCO Combiflash refers to flash chromatography on silica gel using the Isco Combiflash® separation system: RediSep normal phase flash column, flow rate 30-40 ml / min;

(xi) the following abbreviations may be used:

ACN

BINAF

Vosgo kdi

DAST acetonitrile

2,2'-bis (diphenylphosphino) -1,1'-binaphthyl tert-butyloxycarbonyl anhydride

Ν, Ν-carbonyldiimidazole diethylaminosulfur trifluoride dxm

DIPEA / DIEA

DMAc

DMF DMAP dmso ei dichloromethane

A, A-diisopropylethylamine A / A-dimethylacetamide

BUT; A-dimethylformamide 4-dimethylaminopyridine dimethyl sulfoxide enantiomeric excess

EtOAc / EA ethyl acetate

Et ₂ O

GC

HATU diethyl ether gas chromatography

O- (7-azabenzotriazol-1-yl) -A, A, L ^p , Ν'-tetramethyluronium hexafluorophosphate at

Hex hexanes HPLC high performance liquid chromatography h / h h

KO'Vp

LCMS mpem-potassium butoxide liquid chromatography-mass spectrometry

LDA lithium diisopropylamide

MeCN acetonitrile

MeOH methanol

min minutes MTBE methyl tert-butyl ether in tech. nights during the night Pd ₂ (dba) ₃ PE tris (dibenzylideiacetone) dialladium (0) petroleum ether / RgON isopropyl alcohol rac. racemic

- 31 037916

TBAF tetra-n-butylammonium fluoride TEA triethylamine TFA trifluoroacetic acid THF tetrahydrofuran TMS trimethylsilyl Tozil, Ts para-toluenesulfonyl svezhh-ms ultra high performance liquid chromatography-mass spectrometry

Conditions for carrying out and testing the nitrosoene reaction

In some aspects, the nitrosoene reaction is used to introduce the desired functionality of the allyl hydroxylamine. (Waldemar Adam and Oliver Krebs, Chem. Rev., 2003, 103, 4131-4146; Charles P. Frazier, Jarred R. Engelking, and Javier Read de Alaniz, J. Am. Chem. Soc, 2011, 133 (27) , 10430-10433; Leoni I. Palmer, Charles P. Frazier, Javier Read de Alaniz, Synthesis 2014, 46, 269-280). In the presence of an oxidizing agent, hydroxylamines are oxidized to form highly reactive nitroso compounds that react with an allyl substrate, as shown in Scheme 10. The oxidizing agent can be an organic oxidizing agent or a combination of an oxidizing agent, a metal catalyst and, optionally, an amine additive. Nitroso compounds can be formed in situ (as in the reaction to form a compound of formula VI) from hydroxylamine, or prepared separately and then added to a substrate, for example, a compound of formula XI. In the case of a 1,2,3,6-tetrahydropyridine backbone with an N-carbamate functional group, it is believed that the 1,3-allyl stress in the N-Boc functional group causes the R ¹ substituent to assume a pseudoaxial orientation. It is believed to block the approach of the nitrosoreagent from this side of the double bond. Consequently, the nitrosoreagent reacts regio- and diastereoselectively from the opposite side of R ¹ with the formation of the target product with high diastereoselectivity and regioselectivity. Enantioselectivity was measured and was found to be high in all cases, over 99% ee, when analyzed by chiral HPLC.

Scheme 10 pg ₂

NHOH

Oxidizing agent, solvent or catalyst, additive, oxidizing agent, solvent

Substrate Ri Product

Reaction parameters such as oxidant, catalyst, ligand, solvent, stoichiometry of reactants, temperature and reaction time can influence the result of the reaction and have been tested and optimized. Some of the results are summarized in the following tables (N / A = undefined).

- 32 037916

Oxidizer check

Recording Substrate Reagents and conditions Oxidizing agent Conversion (as determined by% HPLC peak area) Product Ref. vba one PGi = PG2 = Boc Ri = CONH ₂ R ₂ = Me, R ₃ = H BocNHOH (1.5 eq.) CuCl (0.05 equiv.) Pyridine (0.013 equiv.) DXM (10vol.), 17-25 ° C, 20-60 h About ₂ , 1 atm 84.5 5.9 2 FeCl ₃ , 1.2 eq. 15.3 43.2 3 MnO ₂ , 1.2 eq. 48.0 10.28 4 NaIO4, 2.2 eq. 43.8 21.3 five w-SRVA, 2.2 equiv. 10.7 30.9 6 BPO, 1.0 eq. TEMPO, 1.5 equiv. 32.3 19.6 7 NYU ₃ , 1.2 equiv. 6.4 s, o eight 1 ₂ , 1.2 equiv. 5.8 24.6 nine H ₂ O ₂ , 3.0 eq. 78 8.9 10 MnO ₂ , 1.2 eq. H ₂ O ₂ , 1 drop 71.6 5.0 AND IBX, 1.2 equiv. BUT 12 PhI (OAc) ₂ , 1.2 eq. but 13 hydroperite, 1.2 eq. 63.9 12.1 fourteen NCS, 1.0 eq. 59.9 12.5 fifteen DMP, 1.0 eq. BUT sixteen 3% H ₂ O ₂ , 1.2 eq 70 3.1 17 1.5% H ₂ O ₂ , 1.2 eq 64.4 4D 18 6% H ₂ O ₂ , 1.2 eq 73.6 5.5 nineteen NMM0, 1.0 eq. 37.8 38.6 twenty PGi = PG ₂ = Boc Ri = CO ₂ Me R ₂ = CH ₂ OPMB, R ₃ = H BocNHOH (1.5 eq.) CuCl (0.05 equiv.) Pyridine (0.013 equiv.) DXM (10vol.), 17-25 ° C, 20-60 h About ₂ , 1 atm. 36% of isolated product 21 PGi = PG ₂ = Boc Ri = CONH ₂ R ₂ = H, R ₃ = Me BocNHOH (1.5 eq.) CuCl (0.05 equiv.) Pyridine (0.013 equiv.) DXM (10vol.), 20-30 ° C, 20-60 h About ₂ , 1 atm. 46.08 10.28 22 FeCl ₃ (1.2 eq.) 1.33 71.43 23 MnO ₂ (1.2 equiv.) 40.79 16.85 24 NaIO ₄ (1.2 equiv.) 15.2 35.58 25 w-CPVA (1.2 equiv.) 3.04 89.1 26 NYU ₄ (1.2 equiv.) 0.52 46.47 27 30% H ₂ O ₂ (1.2 eq.) 4.69 7.9 28 1 ₂ (1.2 equiv.) 1.39 52.85 29 CaO ₂ (1.2 equiv.) 19.66 33.73 thirty BocNHOH (3.0 eq.), CuCl (0.05 eq.), Sulfolane (5 vol.), H ₂ O (5 vol.), Py (0.13 eq.), 0-5 ° C, H ₂ O ₂ (3%, 3 eq.), 16 h H ₂ O ₂ (3%, 3 eq.), 96 h 54.3 38.8 30.5 0.24 31 BocNHOH (2.0 eq.), CuBrMe ₂ S (0.05 eq.), Sulfolane (5 vol.), H ₂ O (0.5 vol.), Py (0.13 eq.), 25-30 ° C Н ₂ О ₂ (3%, 2 equiv.), 64 h 70.0 4.1

- 33 037916

Solvent check

Recording Substrate Reagents and conditions Solvent, time Conversion (as determined by HPLC peak area%) Product Ref. vba one PGi = PG2 = Boc Ri = CONH ₂ R ₂ = Me, R ₃ = H BocNHOH (l, 5 equiv.) CuCl (0.05 equiv.) Pyridine (0.013 equiv.), 15 ° C, air 5% THF in DXM (10 vol.), 90 h 77.6 5.0 2 10% THF in DXM (10 vol.), 90 h 77.5 4.7 3 20% THF in DXM (10 vol.), 90 h 68.8 19.4 4 DXM (10 vol.), 68 h 65.8 20.4 five THF (10 vol.), 20 h 81.9 7.8 6 EYU As (10 rev.), 68 h 69.5 19.0 7 Acetone (10 vol.), 68 h 87.3 3.6 eight PGi = PG ₂ = Boc Ri = CONH ₂ R ₂ = H, R ₃ = Me BocNHOH (l, 5 eq.) CuCl (0.05 equiv.) Pyridine (0.013 equiv.), 20-30 ° C, air, 60 h DXM (10 vol.) 35.2 20.6 nine DXM / H ₂ O 1: 1 (10 vol.) 38.0 18.7 10 MeON (Yoob) 50.0 21.8 AND MeOH / H ₂ O 1: 1 (10 vol.) 45.7 28.3 12 EYUN (10ob.) 47.5 15.9 13 z-PrOH (10 vol.) 43.3 10.0 fourteen 2-methyl-2-butanol (10 vol.) 48.0 16.6 fifteen Ethylene glycol (10 vol.) 14.1 76.4 sixteen Polyethylene glycol 2000 / NYU (5 g / 5 vol.) 9.3 96.9 17 sulfolane / H ₂ O 1: 1 (10 vol.) 57.0 14.7 18 sulfolane / H ₂ O 9: 1 (10 vol.) 55.58 17.5 nineteen sulfolane / H ₂ O 3: 7 (10 vol.) 23.3% 66.7 twenty sulfolane / H ₂ O 1: 1 (5 vol.) 62.3 5.9 21 BocNHOH (l, 5 equiv.) CuCl (0.05 equiv.) Pyridine (0.013 equiv.), 15-30 ° C, О ₂ (1 atm.), 48 h dmfa / n ₂ about 5 about. / 5 about. 58.11 1.71 22 nmp / h ₂ o 5 rpm / 5 rpm 44.18 0.43

- 34 037916

Checking the catalyst

Recording Substrate Reagents and conditions Catalyst Conversion (as determined by HPLC peak area%) Product Ref. vba one PGi = PG ₂ = Boc Ri = CONH ₂ R ₂ = H, R ₃ = Me BocNHOH (l, 5 equiv.) CuCl (0.05 equiv.) Pyridine (0.013 equiv.) DCM (10 vol.), 20-30 ° C, О ₂ (1 atm), 65 h CuCl, 0.05 eq. 35.2 20.6 2 CuBr, 0.05 equiv. 30.7 25.2 3 Cui, 0.05 equiv. 21.5 43.1 4 Cu (OAc) ₂ , 0.05 eq 33.0 28.5 five sulfolane / H ₂ O 5 vol. / 5 vol., 24 h 58.49 1.30 6 sulfolane / H ₂ O 5 vol. / 5 vol., 96 h 38.83 0.24 7 HOAc / H ₂ O 5 vol. / 5 vol., 96 h 7.95 71.02 eight BocNHOH (l, 5 eq.), Pyridine (0.013 eq.), Sulfolane (5 vol.), H ₂ O (5 vol.), O ₂ , 20-30 ° C, CuCl, 0.05 equiv., 24 h 58.49 1.30 nine CuCl, 0.05 equiv., 96 h 38.83 0.24 10 CuBr, 0.05 equiv. 72 h 63.42 7.31 AND Cui, 0.05 equiv. 67.49 7.35 12 CuSCN (0.05 equiv.), 72 h 32.98 47.05 13 Cu (OAc) 2 (0.05 equiv.), 72 h 59.69 15.94 fourteen CuBr-SMe ₂ (0.05 equiv.), 72 h 68.71 3.36 fifteen CuCN (0.05 equiv.), 84 h 5.60 5.32 sixteen Cu (CH ₃ CN) ₄ pf ₆ (0.05 equiv.), 84 h 61.56 14.07 17 Cu (OT1) -toluene (0.05 eq.), 84 h 66.24 9.31 18 CuCl (0.05 equiv.) 2-ethyloxazoline (0.05 equiv.), 84 h 59.03 15.20 nineteen CuCl (0.05 equiv.) TBAI (0.05 equiv.), 89 h 49.9 26.26

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Checking additives

Recording Substrate Reagents and conditions Ligand Transformation (as defined by % HPLC peak area) Product Ref. in-va one PGi = PG ₂ = Boc Ri = CONH 2 r ₂ = h, R ₃ = Me BocNHO H (1.5 equiv.) CuCl (0.05 equiv.) DCM (10 vol.), 20-30 ° C, О ₂ (1 atm), 65 h Pyridine (0.013 equiv.) 35.2 20.6 2 Ethane-1,2-diamine (0.013 equiv.) 31.8 29.0 3 (17?, 27?) - cyclohexane-1,2-diamine (0.013 equiv.) 36.3 30.1 4 N1, A7-dimethylethane-1,2-diamine (0.013 equiv.) 35.0 21.5 five 2,6-di-tert-butyl-4-methylpyridine (0.013 equiv.) 31.5 34.0 6 1,10-o-phenanthroline (0.013 equiv.) but 7 Ligand free 24.8 47.4 eight BocNHO H (1.5 eq.), CuCl (0.05 eq.), Sulfolan n (5 vol.), H ₂ O (5 vol.), O ₂ (1 atm), 2030 ° C h ₂ n ^ \. _in ^NH 2,0,013 eq, 72 h 37.73 27.4 9 nine 1 0.013 eq, 72 h 5.60 5.32 10 1,10-o-phenanthroline (0.013 equiv.), 72 h 64.45 5.14 AND 1 I 0.013 equiv. 96 h 69.09 0.94 12 ΑΑ _Ν || ₂ trans - 0.013 eq. 96 h 69.11 0.93 13 Pyridine (0.013 equiv.), 24 h 58.49 1.30 fourteen Pyridine (0.013 equiv.), 96 h 38.83 0.24 fifteen Pyridine (0.065 equiv.), 84 h 74.1 7.1 sixteen Pyridine (0.13 equiv.), 24 h 73.80 5.57 17 ΑΑ _Ν || ₂ trans - 0.013 eq, 65 h 66.63 5.09 18 ΝΗ ₂ ΑΧχι ι ₂ trans - 0.13 equiv, 48 h 9.48 86.4 7 nineteen 1 ⁿ 0.013 eq 72 h 62.64 10.4 five

- 36 037916

twenty 0.013 equiv., 72 h 64.43 9.24 21 0.013 equiv., 72 h 66.21 7.52 22 Cis - 0.013 equiv., 72 h 65.09 6.79 23 0.013 equiv., 130 h 71.2 10.0 24 0.013 equiv., 130 h 66.3 12.0 25 0.013 equiv., 130 h 15.0 61.8 26 DMAP, 0.03 equiv., 60 h 70.88 5.91 0.013 equiv., 60 h 71.56 4.93 27 0.013 equiv., 89 h 57.34 I, 8 5 28 0.013 equiv., 89 h 61.04 14.6 5 29 18-crown-6 (0.001 equiv.), KC1 (0.05 equiv.), 36 h 73.67 5.53 thirty Sodium L-ascorbate (0.1 eq.), 36 h Next qty 98.7 2 31 18-crown-6 (0.001 equiv.), KC1 (0.05 equiv.), Py (0.013 equiv.), 86 h 75.8 4.8 32 0.013 eq., 18-crown-6 (0.001 eq.), KC1 (0.05 eq.), 86 h 74.0 10.6

- 37 037916

33 '-o 0.013 equiv., 18-crown -6 (0.001 equiv.), KC1 (0.05 equiv.), 86 h 60.6 22.5 34 BocNHO H (1.5 eq.), CuCl (0.05 eq.), Sulfolan n (5 vol.), H ₂ O (5 vol.), Air, 20-30 ° C Ru (0.013 eq.), 85 h 46.74 24.5 5 35 H H H ₂ N ^^ nh ₂ Η 0.013 equiv., 85 h 54.08 16.8 1 36 О О А А Η ₂ Ν Ν ΝΗ ₂ Η 0.013 equiv., 85 h 51.32 18.7 8 37 H Η 0.013 equiv., 85 h 53.35 17D 38 BUT VV 0.013 equiv., 85 h 51.11 18.2 one 39 A WV A A 0.013 equiv., 85 h 55.61 14.3 9 40 BocNHO H (1.5 eq.), CuBrMe ₂ S (0.05 eq.), Sulfol n (5 vol.), H ₂ O (5 vol.), O ₂ , 20-30 ° C Pyridine (0.013 equiv.), 72 h 68.71 3.36 41 DMAP (0.013 equiv.), 86 h 66.99 10.8 8 42 one 1 0.013 equiv., 86 g 3.01 96.5 7 43 nh ₂ AA _N || ₂ trans - 0.013 equiv., 86 h 71.53 6.53 44 BocNHO H (3 eq.), CuCl (0.05 eq.), Sulfolan n (5 vol.), H ₂ O (5 vol.), H ₂ O ₂ (3%, 3 NH ₂ | ₂ trans - 0.013 eq, 66 h 74.3 8.7 eq.), 20-30 ° C

After checking the best conditions for the substrate with PG1 = PG2 = Boc, R1 = CONH2, R2 = Me, R ₃ = H turned out to be: BocNHOH (1.5 equiv.), CuCl (0.05 equiv.), Py (0.013 equiv. ), DCM (10 vol), O2 (1 atm.), 15-25 ° C. For a substrate with PG1 = PG2 = Boc, R1 = CONH2, R ₂ = H, R ₃ = Me, the best conditions were: BocNHOH (1.5 equiv.), CuCl (0.05 equiv.), Py (0.013 equiv. ), sulfolane (5 vol.), H ₂ O (5 vol.), O ₂ (1 atm.), 15-25 ° C or BocNHOH (2 eq.), CuBr-Me ₂ S (0.05 eq. ), Py (0.13 equiv.), Sulfolane (5 vol.), H ₂ O (0.5 vol.), H ₂ O ₂ (3% in water, 2-3 equiv.), 1525 ° C.

The following experimental procedures are for illustrative purposes.

BocNHOH (2 equiv.),

CuBr-Me ₂ S (0.05 o θ eq.), Ru (0.13 eq.), A. / \ / _n sulfolane (5 vol.), Η ₂ Ν [

BoA H ₂ O (0.5 vol.) Vos

Н ₂ О ₂ (3% in water, 2 eq.), 15-25 ° C, 29 h

To a mixture of starting material (1 eq.), BocNHOH (2 eq.), CuBr ₂ -SMe ₂ (0.05 eq.) Was added sulfolane (5 vol.) And H2O (0.5 vol.), Pyridine (0, 13 equiv.). The mixture was stirred for 30-40 minutes at 15-25 ° C. 3% H ₂ O ₂ (2 eq.) Was added dropwise over 24-30 h. After the reaction was considered complete, a solution of EDTA-2Na (0.31-0.32 wt. Eq.) In water ( 3-3.2 times the mass) and MTBE (7.7 times the mass). The resulting mixture was stirred for 20-30 minutes and allowed to settle for 20-30 minutes. Two phases

- 38 037916

shared. The aqueous phase was extracted three times with MTBE (4 times the mass). The combined organic solution was dried over Na2SO4 and filtered, concentrated and analyzed by assay method. Yield 47.5%, purity 74.12% as determined by HPLC% peak area.

About I

L L

BocNHOH (1 5 eq.) CuCI (0.05 eq.) Η ₂ Ν | η * ^ N. As, Vos

Py (0.013 equiv.), О ₂ (g) Boc N

DXM OH

CuCl (0.05 equiv.), BocNHOH (1.5 equiv.) And Py (0.013 equiv.) Were added to the crude solution of the substrate from the previous step in DCM. The mixture was stirred under O2 at 20 ± 5 ° C until the amount of starting material was <5% as determined by HPLC% peak area. A solution of EDTA-2Na (5.0 vol.) Was loaded and the resulting mixture was stirred for at least 4 h at 25 ± 5 ° C. The two phases were separated. The aqueous phase was extracted twice with DCM (3.0 vol.). The organic phases were combined and washed once with water (5.0 vol.), Concentrated in vacuo at <40 ° C to about 3.0 original volumes. The reactor was charged with i-PrOAc (5.0 vol.) And the mixture was concentrated in vacuo at <40 ° C to about 4.0 original volumes. This process was repeated one more time. N-heptane (5.0 vol.) Was added to the reactor at 40 ± 5 ° C. The resulting mixture was gradually cooled to 20 ± 5 ° C. The solid was obtained by centrifugation and washed with i-PrOAc / n-heptane (1: 1.2 vol.) And dried under vacuum at 35 ± 5 ° C for at least 12 hours. Yield over 2 steps: 71.68 %, purity 99.7% as determined by HPLC% peak area.

Intermediate 1. (S) -2- (1-Hydroxybut-3-en-2-yl) isoindoline-1,3-dione

CaA O

A 2 L reaction flask containing a magnetic stir bar and sodium carbonate (1.981 g, 18.69 mmol) was placed under high vacuum and dried with an air jet dryer for 10 minutes. After cooling, the flask was filled with nitrogen. Allylpalladium chloride dimer (0.553 g, 1.53 mmol), (1R, 2R) - (+) - 1,2-diaminocyclohexane-N, N'-bis (2-diphenylphosphino-1-naphthoyl) (CAS 174810 -09-4) (3.36 g, 4.25 mmol) and phthalimide (50 g, 339.83 mmol). The flask was then purged with nitrogen for 10 minutes. Then 1.4 L of methylene chloride was added, which had undergone preliminary degassing with nitrogen for ten minutes. This suspension was placed under a nitrogen atmosphere; it was alternately stirred and sonicated for ten minutes to facilitate solvation. At this point, it was a yellow or light orange solution containing a white solid. To this mixture was added 2-vinyloxirane (24.06 g, 343.23 mmol). The resulting mixture was stirred under nitrogen at ambient temperature for approximately 48 hours. LCMS and TLC analysis (1: 1 hexanes: ethyl acetate) during this process indicated the development of the reaction, and final analyzes using these methods indicated complete conversion starting material into one main product. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The yellow viscous liquid was injected onto a 330 g silica gel column: a minimum volume of methylene chloride was used to dilute the crude material. Silica gel chromatography (15-75% ethyl acetate in hexanes, 40 min, 330 g column) was used to isolate the desired product as a viscous yellow liquid that became a pale yellowish white solid (69.6 g, 94%) within several hours under reduced pressure. Optical rotation: (2.02 g / 100 ml, methylene chloride) RR = -72.2, resulting value = -71.

Intermediate 2. (S) -2- (1- (tert-Butyldimethylsilyloxy) but-3-en-2-yl) isoindoline 1,3-dione τββο'Ά'Ά

To a stirred solution of (S) -2- (1-hydroxybut-3-en-2-yl) isoindoline-1,3-dione (intermediate 1, 69.4 g, 319.49 mmol) and imidazole (26.1 g, 383.39 mmol) in methylene chloride (160 mL) at ambient temperature under nitrogen, tert-butyldimethylchlorosilane (55.4 g, 367.41 mmol) was added as a solid. This addition was carried out for approximately ten minutes. Heating of the mixture was observed during this addition. After stirring for two hours, the solution was poured into saturated aqueous sodium bicarbonate solution (approximately 150 ml); this biphasic mixture was shaken and the organic layer was separated. The aqueous layer was back-extracted three times using 200 ml of methylene chloride each time. The organic layers were combined, dried over magnesium sulfate, filtered and concentrated in vacuo. The target product was obtained in the form of a pale yellow

39 037916 solid after drying overnight under high vacuum (107 g, 101%).

Intermediate 3. (S) -1- (tert-Butyldimethylsilyloxy) but-3-en-2-amine TBSO ^^ y ^ nh ₂

To a stirred solution of (S) -2- (1- (tert-butyldimethylsilyloxy) but-3-en-2-yl) isoindoline 1,3-dione (Intermediate 2, 108.28 g, 326.65 mmol) in methanol ( 1 L) hydrazine (35.9 mL, 1143.29 mmol) was added at ambient temperature under nitrogen. The yellow solution was heated to 65 ° C. Thirty minutes after reaching the reaction temperature, a white precipitate was observed in the reaction mixture; this solid quickly became the bulk of the mixture at which time water (about 150 ml) was added to the reaction mixture. Stirring of the reaction mixture was continued without interruption and the solid dissolved within a few minutes. After complete conversion, as determined by LCMS analysis (both starting material and product give strong UV signals and are easily identified by LCMS), heating was stopped and more water was added (total water 600 ml). The mixture was allowed to reach ambient temperature.

Methanol was removed in vacuo at 35 ° C (moderately low pressure); the vacuum was removed and the aqueous solution was heated to approximately 50 ° C, and then extracted with 4x200 ml of methylene chloride. The organic extracts were combined, washed with saturated sodium bicarbonate (aq), washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo at a temperature not exceeding 30 ° C. The target product was obtained as a yellow liquid (58.5 g, 94%).

Intermediate 4.2-Bromo-N-methoxy-N-methylacetamide

OMe about

A stirred solution of potassium carbonate (343 g, 2.48 mol) in water (about 800 ml) was prepared and cooled in an ice bath for 15 min under nitrogen. To this was added O, N-dimethylhydroxylamine hydrochloride (110 g, 1.13 mol) and diethyl ether (about 800 ml). Bromoacetyl bromide (273 g, 1.35 mol) was then added to this mixture using a dropping funnel over twenty minutes. The ice bath was removed and the mixture was stirred under nitrogen atmosphere for 2 hours. The layers were separated and the aqueous layer was extracted with ether (about 350 ml). The organic layers were combined, dried over magnesium sulfate, filtered and concentrated in vacuo. The target product was obtained as a yellow liquid (143 g, 70%).

Intermediate 5. (S) -tert-Butyl-1- (tert-butylgimethylsilyloxy) but-3-en-2-yl (2 (methoxy (methyl) amino) -2-oxoethyl) carbamate

TBSO ^ Y ^ k boc η

A suspension of (S) -1- (tert-butylgimethylsilyloxy) but-3-en-2-amine (Intermediate 3, 60.4 g, 300 mmol) and cesium carbonate (103 g, 315 mmol) in acetonitrile (about 700 ml) and water (about 120 ml) and it was stirred in an ice bath under nitrogen atmosphere for 5 minutes. The mixture was biphasic and remained so during the reaction. Then to this mixture was added 2-bromo-N-methoxy-Nmethylacetamide (Intermediate 4, 57.0 g, 285 mmol) via an addition funnel over 10 minutes. The mixture was stirred for two days while maintaining the temperature at about 0 ° C. The mixture was kept in a freezer overnight. Another 0.05 equiv of electrophile was added. To the mixture was added di-tert-butyl dicarbonate (165 ml, 2M solution in THF). The organic layer was separated from the aqueous (TLC showed no product remained in the aqueous layer) and the organic layer was concentrated in vacuo. After chromatography on silica gel (5-55% ethyl acetate in hexanes), divided into 3 batches, the desired product was obtained as a pale yellow oil (80 g, 66%).

Intermediate 6. (S) -tert-Butyl-1- (tert-butylgimethylsilyloxy) but-3-en-2-yl (2oxopent-3-yl) carbamate tbso ^ A ^

A boc

To a solution of (S) -tert-butyl-1 - (tert-butyldimethylsilyloxy) but-3-en-2-yl (2- (methoxy (methyl) amino) 2-oxoethyl) carbamate (Intermediate 5, 32.5 g , 80.73 mmol) in THF (400 ml) under nitrogen atmosphere at 0 ° C was added dropwise prop-1-enylmagnesium bromide (323 ml, 161.45 mmol). The reaction mixture was stirred at 0 ° C for 1 h, then the reaction was stopped by adding 400 ml of 10% citric acid, additionally diluted with 100 ml of water and extracted with ether. The organics were concentrated and the resulting oil was dissolved in ether and washed with water and brine. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (5-20% ethyl acetate / hexanes) gave the title product as a colorless oil (27 g, 87%).

MS: 384 ES + (C ₂₀ H ₃₇ NO ₄ Si). ¹ H NMR (300 MHz, CDCl ₃ ) δ: 0.05 (2, 6H); 0.88 (s, 9H); 1.39-1.47 (m, 9H); 1.90 (m, 3H); 3.80 (m, 2H); 4.05-4.18 (m, 2H); 4.43-4.76 (m, 1H); 5.22 (m, 2H); 5.86 (m, 1H); 6.21 (m, 1H); 6.91 (m, 1H).

Intermediate 7. (S) -tert-Butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -5-oxo-5,6 dihydropyridine-1 (2H) -carboxylate

(S) -tert-butyl-1 - (tert-butyldimethylsilyloxy) but-3-en-2-yl (2-oxopent-3-enyl) carbamate (Intermediate 6, 27.0 g, 70.39 mmol) was dissolved in toluene (650 ml). The solution was purged with nitrogen for 15 min before adding the 2nd generation Howeida-Grubbs catalyst (0.885 g, 1.41 mmol). The reaction mixture was heated under nitrogen atmosphere at 65 ° C. The reaction mixture was concentrated under reduced pressure. Chromatography on silica gel (10-35% ethyl acetate / hexanes) gave the title product as a solid (17.0 g, 70%). Optical rotation: 0.1 g / dl, methylene chloride = -175.

Intermediate 8. (6S) -tert-Butyl-6 - ((tert-butyldimethylsilyloxy) methyl) -5-methyl-3 (trimethylsilyloxy) -5,6-dihydropyridine-1 (2H) -carboxylate

To a suspension of copper (I) iodide (22.31 g, 117.12 mmol) in diethyl ether (250 ml) at 0 ° C, methyllithium (1.6 M in ether) (146 ml, 234.25 mmol ). The suspension was stirred for 45 min at 0 ° C. To the suspension was added dropwise a solution of (S) -tert-butyl-2 - (((tert-butyldimethylsilyl) oxy) methyl) -5-oxo-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 7, 20 g, 58.56 mmol) in diethyl ether (50 ml) at 0 ° C. After the addition was complete, the reaction mixture was stirred for 45 min at 0 ° C. Chlorotrimethylsilane (1M in THF) (117 ml, 117.12 mmol) was then added dropwise to the reaction mixture, followed by triethylamine (16.28 ml, 117.12 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 2 hours. The reaction mixture was then diluted with ethyl acetate and washed three times with ice-cold saturated sodium bicarbonate solution (added with great care) and then with brine. The organics were dried over sodium sulfate, filtered and concentrated to give a brown oil.

Intermediate 9. (S) -tert-butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -3-methyl-5oxo-5,6-dihydropyridine-1 (2H) -carboxylate

To a solution of (6S) -tert-butyl-6 - (((tert-butyldimethylsilyl) oxy) methyl) -5-methyl-3 - ((trimethylsilyl) oxy) -5,6-dihydropyridine-1 (2H) -carboxylate ( Intermediate 8, 24.1 g, 56.08 mmol) in acetonitrile (280 mL) was added palladium (II) acetate (12.59 g, 56.08 mmol) at room temperature. The reaction mixture was stirred at room temperature for about 40 hours, then diluted with ethyl acetate and filtered through celite. The filtrate was concentrated on silica gel. After chromatography on silica gel (0% -20% ethyl acetate / hexanes), (S) -tert-butyl-2 - (((tert-butyldimethylsilyl) oxy) methyl) -3-methyl-5-oxo-5,6-dihydropyridine- 1 (2H) -carboxylate (12.95 g, 65%) as a yellow solid.

Intermediate 10. (2S, 5S) -tert-butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -5hydroxy-3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

To a suspension of cerium (III) chloride (8.98 g, 36.42 mmol) and (S) -tert-butyl-2 - (((tert-butyldimethylsilyl) oxy) methyl) -3-methyl-5-oxo-5,6 α-dihydropyridine-1 (2H) -carboxylate (Intermediate 9, 12.95 g, 36.42 mmol) in methanol (200 ml) at 0 ° C. sodium borohydride (1.387 g, 36.42 mmol) was added in portions. After 15 minutes, the reaction mixture was diluted with saturated ammonium chloride (100 ml) and water (100 ml), then extracted twice with ether. The organic extracts were washed with brine, dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0-20% ethyl acetate / hexanes), (2S, 5S) -tert-butyl-2 - (((tert-41 037916 butyldimethylsilyl) oxy) methyl) -5-hydroxy-3-methyl-5 , 6-dihydropyridine-1 (2H) -carboxylate (9.79 g,

75%) as a colorless oil.

Intermediate 11. (2S, 5R) -tert-Butyl-5- (N- (αlyloxy) -2-nitrophenylsulfonamido) 2 - ((tert-butyldimethylsilyloxy) methyl) -3-methyl-5,6-dihydropyridine-1 ( 2H) -carboxylate

To a solution of (2S, 5S) -tert-butyl-2 - (((tert-butyldimethylsilyl) oxy) methyl) -5-hydroxy-3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 10, 9 , 79 g, 27.38 mmol) in toluene (100 ml) triphenylphosphine (8.58 g, 32.86 mmol), N- (αlyloxy) -2nitrobenzenesulfonamide (7.07 g, 27.38 mmol) were added at room temperature and diisopropyl azodicarboxylate (6.47 ml, 32.86 mmol). The reaction mixture was stirred overnight, then filtered and concentrated. The resulting oil was purified twice with hexanes and filtered. After chromatography on silica gel (0-20% ethyl acetate / hexanes), (2S, 5R) -tert-butyl-5- (N- (allyloxy) -2-nitrophenylsulfonamido) -2 - (((tert-butyldimethylsilyl) oxy) methyl) -3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate (10.75 g, 66%) as a light yellow foam.

Intermediate 12. (2S, 5R) -tert-Butyl 5- (N- (allyloxy) -2-nitrophenylsulfonamido) 2- (hydroxymethyl) -3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

To a solution of (2S, 5R) -tert-butyl-5- (N- (αlyloxy) -2-nitrophenylsulfonamido) -2 - (((tert-butyldimethylsilyl) oxy) methyl) -3-methyl-5,6-dihydropyridine- 1 (2H) -carboxylate (Intermediate 11, 10.75 g, 17.98 mmol) in THF (100 ml) at 0 ° C was added tetrabutylammonium fluoride (1M in THF) (23.38 ml, 23.38 mmol) ... The reaction mixture turned from yellow to greenish brown. The reaction mixture was stirred for approximately 2 hours, then concentrated on silica gel. After chromatography on silica gel (0-70% ethyl acetate / hexanes), (2S, 5R) -tert-butyl-5- (N (allyloxy) -2-nitrophenylsulfonamido) -2- (hydroxymethyl) -3-methyl-5, 6-dihydropyridine-1 (2H) carboxylate (7.72 g, 89%) as a brownish-beige foam.

Intermediate 13. (2S, 5R) -5- (N- (Allyloxy) -2-nitrophenylsulfonamido) -1- (tert-butoxycarbonyl) -3-methyl-1,2,5,6-tetrahydropyridine-2-carboxylic acid

To a solution of iodic acid (1.588 g, 8.27 mmol) in wet acetonitrile (20 ml) (0.75 vol% water) at room temperature was added chromium (VI) oxide (0.019 g, 0.19 mmol). The mixture was stirred until complete dissolution. To a solution of (2S, 5R) -tert-butyl 5- (N- (αlyloxy) -2-nitrophenylsulfonamido) -2 (hydroxymethyl) -3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 12, 2 g, 4.14 mmol) in wet acetonitrile (20 ml) (0.75 vol.%) At 0 ° С, the previously obtained iodic acid / chromium oxide solution (20 ml, 2 eq.) Was added dropwise and stirred within 15 minutes. The reaction mixture was diluted with DCM (100 ml) and washed with 10% citric acid (50 ml) and twice with brine. The organics were dried over sodium sulfate, filtered and concentrated to give a brownish beige foam (1.9 g, 92%).

Intermediate 14. (2S, 5R) -tert-Butyl 5- (N- (allyloxy) -2-nitrophenylsulfonamido) 2-carbamoyl-3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

To a solution of (2S, 5R) -5- (N- (αlyloxy) -2-nitrophenylsulfonamido) -1- (tert-butoxycarbonyl) -3methyl-1,2,5,6-tetrahydropyridine-2-carboxylic acid (Intermediate 13 , 1.9 g, 3.82 mmol) in DMF (9.5 ml) at 0 ° C was added dropwise HATU (2.178 g, 5.73 mmol), ammonium chloride (0.613 g, 11.46 mmol) and DIEA (2.67 ml, 15.28 mmol). The reaction mixture was warmed to room temperature and stirred for 15 minutes. The reaction mixture was diluted with ethyl acetate and washed with saturated sodium bicarbonate and a 1: 1 mixture of brine and water. After chromatography on silica gel (0-70% ethyl acetate / hexanes), (2S, 5R) -tert-butyl-5- (N- (allyloxy) -2nitrophenylsulfonamido) -2- (hydroxymethyl) -3-methyl-5,6 β-dihydropyridine-1 (2H) -carboxylate (1.270 g, 67%) as a light orange foam.

- 42 037916

Intermediate 15. (2S, 5R) -tert-Butyl-5 - ((allyloxy) amino) -2-carbamoyl-3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

To a solution of (2S, 5R) -tert-butyl 5- (N- (allyloxy) -2-nitrophenylsulfonamido) -2-carbamoyl-3methyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 14, 3 , 63 g, 7.31 mmol) in acetonitrile (100 ml) at room temperature were added potassium carbonate (5.05 g, 36.55 mmol) and thiophenol (3.75 ml, 36.55 mmol). The reaction mixture was stirred overnight at room temperature. The reaction mixture was concentrated and the resulting residue was purified by washing with DCM and filtered to remove solids. The filtrate was concentrated on silica and purified. After chromatography on silica gel (0-90% ethyl acetate / hexanes), (2S, 5R) -tert-butyl-5 - ((allyloxy) amino) -2-carbamoyl-3-methyl-5,6-dihydropyridine-1 (2H) β-carboxylate (1.49 g, 65%) as a yellow oil.

Intermediate 16. (2S, 5R) -tert-Butyl 5- (N- (allyloxy) -1H-imidazole-1carboxamido) -2-carbamoyl-3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

To a solution of (2S, 5R) -tert-butyl-5 - ((allyloxy) amino) -2-carbamoyl-3-methyl-5,6-dihydropyridine 1 (2H) -carboxylate (Intermediate 15, 1.49 g, 4 , 79 mmol) in THF (30 ml) at room temperature were added N, N-diisopropylethylamine (2.5 ml, 14.36 mmol) and N, N-carbonyldiimidazole (2.328 g, 14.36 mmol). The reaction mixture was stirred for about 2 hours at room temperature. Another equivalent of CDI was added and the reaction was stirred at room temperature for 1 h, then another equivalent of CDI was added and the reaction was stirred for an additional 1 h. The reaction mixture was diluted with DCM and washed four times with a 1: 1 mixture of brine and water, then dried over magnesium sulfate, filtered and concentrated to an off-white foam, 1.86 g.

Intermediate 17. (2S, 5R) -6- (Allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-en-2-carboxamide

To a solution of (2S, 5R) -tert-butyl 5- (N- (allyloxy) -1H-imidazole-1-carboxamido) -2-carbamoyl-3methyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate compound 16, 1.86 g, 4.59 mmol) in DCM (20 ml) at 0 ° C was added trifluoroacetic acid (3.53 ml, 45.88 mmol). The reaction mixture was allowed to slowly warm to room temperature and stirred overnight. The reaction mixture was concentrated. The oil was redissolved in DCM and washed with saturated sodium bicarbonate solution. The aqueous layer was extracted once with about 10% MeOH / DCM. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0-30% acetone / dichloromethane), (2S, 5R) -6- (allyloxy) -3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2 β-carboxamide (0.83 g, 76%) as a light yellow oil.

Example 1. (R) -Ethyl-2 - ((2S, n5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yloxy) -2 -fluoroacetate

Example 2. (S) -Ethyl-2 - ((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yloxy) -2 -fluoroacetate

Example 3. (2S) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy}

- 43 037916 (fluorine) ethanic acid lithium salt

Example 4. (2R) - {[(2 S, 5R) -2-Carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy} (fluoro ) ethanic acid lithium salt

Example 5. {[(2S, 5R) -2-Carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} (fluoro) acetic acid lithium salt (mixture of diastereomers)

Examples 1-2.

To a solution of (2S, 5R) -6- (allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxamide (Intermediate 17, 100 mg, 0 , 42 mmol) in methanol (3 ml) at room temperature were added 1,3-dimethylbarbituric acid (132 mg, 0.84 mmol) and tetrakis (triphenylphosphine) palladium (0) (48.7 mg, 0.04 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to give an orange film. The orange film was dissolved in DMF (3 ml) and potassium carbonate (175 mg, 1.26 mmol) and ethyl bromofluoroacetate (0.299 ml, 2.53 mmol) were added. The reaction mixture was stirred overnight at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0-70% ethyl acetate / hexanes), ethyl 2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct -3-en-6-yl) oxy) -2-fluoroacetate (97 mg, 76%) as a mixture of the diastereomers of Example 1 and Example 2, as an orange foam. MS: 198 ES + (C ₈ H _n N ₃ O ₃ ).

Examples 3-5.

To a solution of ethyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2 -fluoroacetate (examples 1-2, 97 mg, 0.32 mmol) in THF (3 ml) and water (1 ml) at -5 ° C was added lithium hydroxide (9.25 mg, 0.39 mmol) in the form of aqueous solution (0.5 ml). The mixture was stirred at 0 ° C for 60 min, allowed to warm to room temperature and stirred for 15 min. The pH of the reaction mixture was adjusted to pH 7 with 0.5 M HCl. THF was evaporated, after which the remaining aqueous phase was frozen and lyophilized.

Cleaning up examples 1-5. The mixture obtained from the reaction described in Examples 3-5 was purified using reverse phase HPLC (Synergi Polar RP 21.2 mm χ 100 mm, 4 μm, conjugated with YMC C30 20 mm χ 150 mm, 5 μm; 0 % -50% acetonitrile in water, 10 min; 20 ml / min) to obtain:

Example 1: ethyl - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} (fluoro) acetate (first ester eluted): 4.4 mg, 4.5%. MS: 302 ES + (C ₁₂ H ₁₆ FN ₃ O ₅ ). 1H NMR (300 MHz, DMSO-d6) δ: 1.21 (t, 3H); 1.62 (s, 3H); 3.05 (m, 1H); 3.75 (m, 1H); 4.03 (m, 1H); 4.20 (m, 3H); 6.01 (m, 1H); 6.13-6.31 (d 1H); 7.36 (bs, 1H); 7.81 (bs, 1H).

Example 2: (S) -ethyl-2 - ((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yloxy) -2 β-fluoroacetate (second eluted ester): 4.2 mg, 4.3% MS: 302 ES + (C ₁₂ H ₁₆ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-dg) δ: 1.26 (t, 3H); 1.63 (s, 3H); 3.08 (m, 1H); 3.75 (m, 1H); 3.94 (m, 1H); 4.20 (m, 1H); 4.27 (q, 2H); 6.03 (m, 1H); 6.24-6.50 (d, 1H); 7.37 (bs, 1H); 7.83 (bs, 1H).

Example 3: (2S) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} ( fluorine) ethanoic acid (first acid to elute): 7.7 mg, 8.8%. MS: 274 ES + (C ₁₀ H ₁₂ FN ₃ O ₅ ).

1H NMR (300 MHz. DMSO-dg) δ: 1.61 (s, 3H); 3.05 (m, 1H); 3.68 (m, 1H); 3.96 (m, 1H); 4.13 (m, 1H); 5.12-5.33 (d, 1H); 6.03 (m, 1H); 7.31 (bs, 1H); 7.80 (bs, 1H).

Example 4: (2R) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} ( fluoro) ethanoic acid (second eluted acid): 9.9 mg, 11% MS: 274 ES + (C ₁₀ H ₁₂ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-de) δ: 1.61 (s, 3H); 3.05 (m, 1H); 3.70 (m, 1H); 4.00 (m, 1H); 4.13 (m, 1H); 5.15-5.37 (d, 1H); 6.01 (m, 1H); 7.31 (bs, 1H); 7.78 (bs, 1H).

Example 5: {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy}

- 44 037916 (fluoro) acetic acid (mixture of diastereomers): 20.4 mg, 23% MS: 274 ES + (C ₁₀ H ₁₂ FN ₃ O ₅ ).

NMR (300 MHz, DMSO-d ₆ ) δ: 1.62 (s, 6H); 3.06 (m, 2H); 3.70 (m, 2H); 4.01 (m, 2H); 4.14 (m, 2H); 5.14-5.35 (d, 1H); 5.18-5.40 (d, 1H); 6.03 (m, 2H); 7.32 (bs, 2H); 7.80 (bs, 2H).

The absolute stereochemistry for all compounds was determined by characterizing the structure of the co-crystal of Example 4 complexed with AmpC. The second diastereomer, example 3, was assigned the opposite absolute stereochemistry at the fluoroacetate carbon atom. The stereochemistry of each of the esters was assigned by hydrolysis of each of the esters to the corresponding acid and comparing the retention times in HPLC with the retention times of Examples 3 and 4.

Example 6. Ethyl - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} (fluoro) acetate (mixture of diastereomers)

According to the procedure for examples and 2, (2S, 5R) -6- (allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo

[3.2.1] oct-3-ene-2-carboxamide (intermediate 17, 0.506 g, 2.13 mmol) was converted to 70 mg of Example 6 after HPLC (Synergi Polar RP 21.2 mm x 100 mm, 4 μm, conjugated with YMC C30 20 mm x 150 mm, 5 μm; 20% -60% acetonitrile in water, 10 min; 20 ml / min) and lyophilization. MS: 302 ES + (C12H16FN3O5).

Ή NMR (300 MHz, DMSO-d6) δ: 1.22 (t, 3H); 1.27 (t, 3H); 1.63 (s, 6H); 3.07 (m, 2H); 3.76 (m, 2H); 3.94 (m, 1H); 4.05 (m, 1H); 4.20 (m, 2H); 4.26 (m, 4H); 6.03 (m, 2H); 6.06-6.24 (d, 1H); 6.14-6.32 (d, 1H); 7.37 (bs, 1H); 7.83 (bs, 1H).

Example 7. Ethyl - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} (difluoro) acetate

(2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (interm. 17, 204.2 mg, 0 , 86 mmol) was converted following the procedure for Examples 1 and 2 using ethyl bromodifluoroacetate (0.441 mL, 3.44 mmol) to give 130 mg (47%) of the title compound as an orange foam. As a result of reverse phase chromatography at 40 mg (Synergi Polar RP 21.2 mm x 100 mm, 4 μm, conjugated with YMC C30 20 mm x 150 mm, 5 μm; 20-40% acetonitrile in water, 10 min; 20 ml / min) gave the title compound as a white solid (21 mg). MS: 320 ES + (C ₁₂ H ₁₅ F ₂ N ₃ O ₅ ).

Ή NMR (300 MHz, DMSO-d ₆ ) δ: 1.30 (t, 3H); 1.65 (s, 3H); 3.19 (m, 1H); 3.84 (m, 1H); 4.06 (m, 1H); 4.26 (m, 1H); 4.39 (q, 2H); 6.05 (m, 1H); 7.42 (bs, 1H); 7.87 (bs, 1H).

Example 8. {[(2S, 5R) -2-Carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} (difluoro) acetic acid lithium salt

Ethyl

2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2,2 difluoroacetate ( example 7, 113.7 mg, 0.36 mmol) was converted according to the procedure for examples 35. After HPLC (Synergi Polar RP 21.2 mm x 100 mm, 4 μm, conjugated with YMC C30 20 mm x 150 mm, 5 μm; 100% water, 10 min; 20 ml / min) and lyophilization gave a white solid, 36.7 mg. MS: 292 ES + (C ₁₀ H _n F ₂ N ₃ O ₅ ).

Ή NMR (300 MHz, DMSO-d6) δ: 1.63 (s, 3H); 3.13 (m, 1H); 3.75 (m, 1H); 3.95 (m, 1H); 4.18 (m, 1H); 6.04 (m, 1H); 7.34 (bs, 1H); 7.83 (bs, 1H).

Example 9. Ethyl - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} acetate

o (2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (intermediate

45 037916 exact substance 17, 0.198 g, 0.83 mmol) was converted according to the procedure for Examples 1-2 using ethyl bromoacetate (0.592 ml) to give 147 mg (62%) as an orange solid. As a result of reverse phase chromatography at 41.5 mg (Synergi Polar RP 21.2 mm χ 100 mm, 4 μm, conjugated with YMC C30 20 mm χ 150 mm, 5 μm; 10% -50% acetonitrile in water, 10 min ; 20 ml / min) gave the title compound as a white solid (32 mg). MS: 284 ES + (C12H17N3O5).

Ή NMR (300 MHz, DMSO-d6) δ: 1.22 (t, 3H); 1.61 (s, 3H); 3.00 (m, 1H); 3.68 (m, 1H); 4.05 (m, 1H); 4.13 (m, 1H); 4.16 (q, 2H); 4.37-4.65 (m, 2H); 6.05 (m, 1H); 7.33 (bs, 1H); 7.77 (bs, 1H).

Example 10. {[(2S, 5R) -2-Carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} acetic acid lithium salt

J --- N ₄

Oh Oh ^ \ ^ he

O

Ethyl 2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) acetate (example 9, 105.5 mg, 0.37 mmol) was hydrolyzed following the procedure for Examples 3-5 to give 34 mg (37%) of the title compound after HPLC (Synergi Polar RP 21.2 mm x 100 mm, 4 μm; 0-20% acetonitrile in water, 10 min; 20 ml / min) and lyophilization. MS: 256 ES + ^(C 10 ^H 13 ^N 3 ^O 5).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 1.60 (s, 3H); 2.96 (m, 1H); 3.58 (m, 1H); 3.87 (m, 2H); 4.05 (m, 1H); 4.27 (m, 1H); 6.08 (m, 1H); 7.27 (bs, 1H); 7.74 (bs, 1H).

Example 11 2 - {[(2S, 5R) -2-Carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy} -2fluoropropanoic acid lithium salt (mixture of diastereomers)

o (2S, 5R) -6- (Allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (intermediate 17, 0.101 g, 0, 43 mmol) was converted according to the procedure for Examples 1-2 using methyl 2-bromo-2-fluoropropanoate (0.315 g, 1.70 mmol) to give methyl 2 - (((2S, 5R) -2carbamoyl- 3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoropropanoate (0.105 g, 82%) as an orange foam. Reverse phase chromatography (Synergi Polar RP 21.2 mm χ 100 mm, 4 μm, coupled to YMC C30 20 mm χ 150 mm, 5 μm; 100% water, 10 min; 20 ml / min) gave the title indicated compound as a white solid (19.1 mg, 19%). MS: 302 ES + (C12H16FN3O5).

1H NMR (300 MHz, DMSO-d6) δ: 1.67 (m, 6H); 3.04 (m, 1H); 3.72 (m, 4H); 3.80 (m, 1H); 4.17 (m, 1H); 6.02 (m, 1H); 7.36 (bs, 1h); 7.81 (bs, 1H).

As a result of hydrolysis according to the procedure for Examples 3-5, 19.1 mg of Example 11 was obtained as a white solid after purification using reverse phase chromatography (Synergi Polar RP 21.2 mm χ 100 mm, 4 μm, conjugated with YMC C30 20 mm χ 150 mm, 5 microns; 100% water, 10 min; 20 ml / min). MS: 288 ES + (C _n H ₁₄ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.45 (m, 3H); 1.61 (s, 3H); 3.02 (m, 1H); 3.65 (m, 1H); 3.92-4.09 (m, 1H); 4.11 (m, 1H); 6.04 (m, 1H); 7.32 (m, 1H); 7.80 (m, 1H).

Intermediate 18. Isopropyl-2-bromo-2-fluoroacetate

To a solution of ethyl 2-bromo-2-fluoroacetate (0.639 ml, 5.41 mmol) in hexanes (70 ml) and isopropanol (7 ml) at 0 ° C was added potassium tert-butylate (0.091 g, 0.81 mmol) in two equal parts, with an interval of 5 minutes. The reaction mixture was stirred for 1 h at 0 ° C. The reaction was stopped by the addition of concentrated HCl (7 ml) and the layers were separated. The organics were washed twice with water, dried over magnesium sulfate, filtered and concentrated at 0 ° C to give a colorless oil (0.88 g, 4.42 mmol, 82%). The product containing trace amounts of hexanes was identified by NMR. The product was used in the next stage in its original form.

1H NMR (300 MHz, CDCl ₃ -d) δ: 1.34 (m, 6H); 5.18 (m, 1H); 6.45-6.62 (d, 1H). Link: Tet. Lett. (2000) 791.

Example 12. Propan-2-yl- (2R) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl ] oxy} (fluorine) etanoate

- 46 037916

Example 13 Propan-2-yl- (2S) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl ] oxy} (fluorine) etanoate

Examples 12-13.

(2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 17, 0.103 g, 0.43 mmol) was converted according to the procedure for Examples 1-2 using isopropyl-2-bromo-2-fluoroacetate (Intermediate 18, 0.518 g, 2.60 mmol). As a result of HPLC and lyophilization, 34 of each diastereomer were obtained (Synergi Polar RP 21.2 mm x 100 mm, 4 μm, conjugated with YMC C30 20 mm x 150 mm, 5 μm; 25% -50% acetonitrile in water, 10 min ; 20 ml / min). The stereochemistry of the ester was assigned by hydrolysis of each of the esters to the corresponding acid and comparing the retention times in HPLC with the retention times of Examples 3 and 4.

Example 12 (First Eluted Ester) 33.4 mg, 24%. MS: 316 ES + (C ₁₃ H ₁₈ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.22 (m, 6H); 1.63 (s, 3H); 3.03 (m, 1H); 3.75 (m, 1H); 4.01 (m, 1H); 4.19 (m, 1H); 5.00 (m, 1H); 6.01 (m, 1H); 6.11-6.29 (d, 1H); 7.37 (bs, 1H); 7.81 (bs, 1H).

Example 13 (Second Eluting Ester) 33.6 mg, 25%. MS: 316 ES + (C ₁₃ H ₁₈ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.27 (m, 6H); 1.63 (s, 3H); 3.07 (m, 1H); 3.75 (m, 1H); 3.94 (m, 1H); 4.20 (m, 1H); 5.05 (m, 1H); 6.03 (m, 1H); 6.02-6.21 (d, 1H); 7.37 (bs, 1H); 7.82 (bs, 1H).

Intermediate 19 2,4-Dimethylpentan-3-yl-2-bromo-2-fluoroacetate

By the method described for Intermediate 18 using ethyl 2-bromo-2-fluoroacetate (0.639 ml, 5.41 mmol) and 2,4-dimethyl-3-pentanol (7.05 ml, 50.27 mmol), the title compound as a colorless oil (0.988 g, 3.87 mmol, 71.6%). 1H NMR (300 MHz, CDCl ₃ -d) δ: 0.94 (m, 12H); 2.01 (m, 2H); 4.73 (m, 1H); 6.52-6.69 (d, 1H).

Example 14 2,4-Dimethylpentan-3-yl- (2S) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy} (fluorine) etanoate

Example 15 2,4-Dimethylpentan-3-yl- (2R) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy} (fluorine) etanoate

Examples 14-15.

(2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 17, 0.097 g, 0.41 mmol) was converted according to the procedure for Examples 1-2 using 2,4-dimethylpentan-3-yl-2-bromo-2-fluoroacetate (0.626 g, 2.45 mmol) to give 35 mg of each diastereomer (23 % for each, 46% in total) after HPLC (Synergi Polar RP 21.2 mm x 100 mm, 4 μm, conjugated with YMC C30 20 mm x 150 mm, 5 μm; 40% -70% acetonitrile in water, 10 min; 20 ml / min) and lyophilization. The stereochemistry of the ester was assigned by hydrolysis of each of the esters to the corresponding acid and comparing the retention times in HPLC with the retention times of Examples 3 and 4.

Example 14 (Second eluting peak) 34.5 mg, 23%. MS: 372 ES + (C ₁₇ H ₂₆ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.86 (m, 12H); 1.63 (s, 3H); 1.94 (m, 2H); 3.07 (m, 1H); 3.78 (m, 1H);

- 47 037916

3.98 (m, 1H); 4.23 (m, 1H); 4.62 (m, 1H); 6.04 (m, 1H); 6.15-6.34 (d, 1H); 7.38 (bs, 1H); 7.81 (bs, 1H).

Example 15 (First eluted peak) 35.2 mg, 23%. MS: 372 ES + (C ₁₇ H ₂₆ FN ₃ O ₅ ).

NMR (300 MHz, DMSO-d6) δ: 0.84 (m, 12H); 1.63 (s, 3H); 1.93 (m, 2H); 3.03 (m, 1H); 3.78 (m, 1H); 4.07 (m, 1H); 4.21 (m, 1H); 4.60 (m, 1H); 5.99 (m, 1H); 6.27-6.45 (d 1H); 7.37 (bs, 1H); 7.83 (bs, 1H).

Intermediate 20. Tetrahydro-2H-pyran-4-yl-2-bromo-2-fluoroacetate F

By the method described for Intermediate 18, using ethyl 2-bromo-2-fluoroacetate (0.319 ml, 2.70 mmol) and tetrahydro-2H-pyran-4-ol (2.333 ml, 24.33 mmol), the specified in the title compound as a colorless oil (0.15 g, 0.62 mmol, 29%). MS: ES + 241.2 for C ₇ H ₁₀ BrFO ₃ .

Ή NMR (300 MHz, chloroform-d) δ ppm: 1.47-1.81 (m, 2H) 1.84-2.13 (m, 2H) 3.39-3.68 (m, 2H) 3.754 , 07 (m, 2H) 4.37 (q, J = 7.18 Hz, 1H) 6.37-6.74 (m, 1H).

Example 16. Tetrahydro-2H-pyran-4-yl - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo

[3.2.1] oct-3-en-6-yl] oxy} (fluoro) acetate (mixture of diastereomers)

By the method described for Examples 1-2 using (2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 17.40 mg, 0.17 mmol) and tetrahydro-2H-pyran-4-yl-2-bromo-2-fluoroacetate (Intermediate 20, 312 mg, 1.29 mmol) gave the title compound after purification inversely. -phase ISCO (15.5 g C18 Gold, 0-80% acetonitrile / water) as a light orange solid as a mixture of diastereomers. (5.5 mg, 9%) MS: ES + 358.1 for C ₁₅ H ₂₀ FN ₃ O ₆ .

Ή NMR (300 MHz, chloroform-d) 5 ppm 1.15-1.42 (m, 2H) 1.60 (br. S., 2H) 1.92 (s, 3H) 3.14-3.45 (m, 2H) 3.56 (d, J = 11.52 Hz, 1H) 3.88 - 4.13 (m, 2H) 4.20-4.50 (m, 2H) 5.09 (br. s., 1H) 5.35-5.58 (m, 1H) 5.635.96 (m, 1H) 6.09 (br.s., 1H).

Intermediate 21.2-Methoxyethyl-2-bromo-2-fluoroacetate

F

Βγ ' ^Λ Υ °' - ^ Ό-

O

Following the method outlined for Intermediate 18 using ethyl 2-bromo-2-fluoroacetate (1.278 mL, 10.81 mmol) and 2-methoxyethanol (13.94 mL, 183.79 mmol) gave the title compound in the form of a colorless oil (1.04 g, 4.84 mmol, 44.7%). The NMR spectrum indicated a 3: 1 mixture of product and starting material which was used as starting material in the next step.

NMR (300 MHz, DMSO-d ₆ ) δ: 3.27 (s, 3H); 3.57 (m, 2H); 4.35 (m, 2H); 7.22-7.38 (d, 1H).

Example 17 2-Methoxyethyl - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy} (fluoro) acetate (mixture of diastereomers)

Example 18 2-Methoxyethyl- (2R) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-en-6-yl] oxy } (fluorine) ethanoate

Example 19 2-Methoxyethyl- (2S) - {[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-en-6-yl] oxy } (fluorine) ethanoate

- 48 037916

Example 17-19.

Following the procedure for Examples 1-2, (2S, 5R) -6- (allyloxy) -3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 17, 0.101 g, 0.43 mmol) was converted to the title compound using 2-methoxyethyl-2-bromo-2-fluoroacetate (Intermediate 21, 0.549 g, 2.55 mmol). As a result of HPLC (Synergi Polar RP 21.2 mm x 100 mm, 4 μm, conjugated with YMC C30 20 mm x 150 mm, 5 μm; 30% -50% acetonitrile in water, 10 min; 20 ml / min) and lyophilization gave the following products (where the stereochemistry of the esters was assigned by hydrolysis of each of the esters to the corresponding acid and comparing the retention times in UPLC with the retention times of examples 3 and 4):

Example 17 Mixture of diastereomers: 14.3 mg, 10.1% MS: 332 ES + (C ₁₃ H ₁₈ FN ₃ O ₅ ).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 1.63 (s, 6H); 3.07 (m, 2H); 3.26 (s, 3H); 3.27 (s, 3H); 3.54 (m, 2H); 3.59 (m, 2H); 3.73 (m, 1H); 3.77 (m, 1H); 3.98 (m, 1H); 4.03 (m, 1H); 4.19 (m, 1H); 4.32 (m, 4H); 6.02 (m, 2H); 6.11-6.30 (d, 1H); 6.17-6.35 (d, 1H); 7.37 (bs, 2H); 7.81 (bs, 2H).

Example 18 (First eluted ester): 22 mg, 15.6%, MS: 332 ES + (C ₁₃ H ₁₈ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 1.63 (s, 3H); 3.06 (m, 1H); 3.27 (s, 3H); 3.55 (m, 2H); 3.77 (m, 1H); 4.05 (m, 1H); 4.20 (m, 1H); 4.30 (m, 2H); 6.02 (m, 1H); 6.18-6.36 (d, 1H); 7.37 (bs, 1H); 7.82 (bs, 1H).

Example 19 (Second eluting ester): 22.8 mg, 16.1%, MS: 332 ES + (C ₁₃ H ₁₈ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 1.62 (s, 3H); 3.07 (m, 1H); 3.27 (s, 3H); 3.59 (m, 2H); 3.75 (m, 1H); 3.99 (m, 1H); 4.20 (m, 1H); 4.34 (m, 2H); 6.03 (m, 1H); 6.11-6.30 (d, 1H); 7.37 (bs, 1H); 7.83 (bs, 1H).

Intermediate 22. (S) -Se-butyl-2-bromo-2-fluoroacetate

F about ¹

To a solution of ethyl 2-bromo-2-fluoroacetate (0.319 ml, 2.70 mmol) and (S) -butan-2-ol (3.72 ml, 40.54 mmol) in hexane (35 ml) at 0 ° C was added potassium tert-butoxide (0.061 g, 0.54 mmol) in two equal portions with an interval of 5 min. The reaction mixture was stirred for 2 h at 0 ° C, then overnight at room temperature. The reaction was stopped by the addition of saturated ammonium chloride solution and the layers were separated. The organics were washed three times with water, dried over magnesium sulfate, filtered and concentrated at 0 ° C to give a colorless oil, 0.550 g, 96%.

1H NMR (300 MHz, DMSO-d6) δ: 0.88 (m, 3H); 1.25 (m, 3H); 1.61 (m, 2H); 4.91 (m, 1H); 7.16-7.33 (m, 1H).

Intermediate 23. (R) -Se-butyl-2-bromo-2-fluoroacetate F o ^z

To a solution of ethyl 2-bromo-2-fluoroacetate (0.319 ml, 2.70 mmol) and (R) -butan-2-ol (3.72 ml, 40.54 mmol) in hexane (35 ml) at 0 ° C was added potassium tert-butoxide (0.061 g, 0.54 mmol) in two equal portions with an interval of 5 min. The reaction mixture was stirred for 2 h at 0 ° C, then at room temperature for 2 h. Then (R) -butan-2-ol (3.72 ml, 40.54 mmol) was added again, followed by another 0, 1 eq. potassium tert-butylate. The reaction mixture was stirred at room temperature overnight. 0.1 eq. Was added every 2 hours over 6 hours. potassium tert-butylate. The reaction was stopped by the addition of saturated ammonium chloride solution and the layers were separated. The organics were washed four times with water, dried over magnesium sulfate, filtered and concentrated at 0 ° C to give a colorless oil, 422 mg, 73%.

NMR (300 MHz, DMSO-d ₆ ) δ: 0.88 (m, 3H); 1.25 (m, 3H); 1.61 (m, 2H); 4.91 (m, 1H); 7.16-7.33 (m, 1H).

Example 20. (2R) - (S) -Stor-butyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl) oxy) -2-fluoroacetate

YES

M F

J ---- N. ⁰ '°> °.

⁰ )

Example 21. (2S) - (S) -Seter-butyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl) oxy) -2-fluoroacetate

- 49 037916

Example 20-21.

Received in accordance with the procedure for examples 1-2. To a solution of (2S, 5R) -6- (αlyloxy) -3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxamide (Intermediate 17, 0.196 g, 0.83 mmol) in methanol (5 ml) at room temperature were added 1,3-dimethylbarbituric acid (0.258 g, 1.65 mmol) and tetrakis (triphenylphosphine) palladium (0) (0.095 g, 0.08 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated to give an orange film. The orange film was dissolved in DMF (5 ml) and potassium carbonate (0.343 g, 2.48 mmol) and (S) -s-butyl-2-bromo-2-fluoroacetate (Intermediate 22, 0.528 g, 2.48 mmol ). The reaction mixture was stirred for about 5 hours at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0% -65% ethyl acetate / hexanes), (S) -s-butyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetate (0.218 g, 80%) as a light yellow foam, a mixture of diastereomers with a ratio of about 1: 1. Separation of diastereomers was performed using reverse phase HPLC (Atlantis T3 19 mm x 150 mm, 30-50% acetonitrile in water, 20 ml / min, 15 min). The stereochemistry of the esters was assigned by hydrolysis of each of the esters to the corresponding acid and comparing the retention times in HPLC with the retention times of Examples 3 and 4.

Example 20 (First eluted peak) 84 mg, 31%, MS: 330 ES + (C ₁₄ H ₂₀ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.84 (t, 3H); 1.22 (d, 3H); 1.55 (m, 2H); 1.63 (s, 3H); 3.03 (m, 1H); 3.76 (d, 1H); 4.04 (m, 1H); 4.19 (s, 1H); 4.86 (m, 1H); 6.01 (m, 1H); 6.13-6.31 (m, 1H); 7.37 (bs, 1H); 7.82 (bs, 1H).

Example 21 (Second eluting peak) 85 mg, 31%. MS: 330 ES + (C ₁₄ H ₂₀ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 0.84 (t, 3H); 1.22 (d, 3H); 1.55 (m, 2H); 1.63 (s, 3H); 3.03 (m, 1H); 3.76 (d, 1H); 4.04 (m, 1H); 4.19 (s, 1H); 4.86 (m, 1H); 6.01 (m, 1H); 6.13-6.31 (m, 1H); 7.37 (bs, 1H); 7.82 (bs, 1H).

Example 22. (2R) - (R) -sec-Butyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl) oxy) -2-fluoroacetate ⁰ I

G ⁰

Example 23. (2S) - (R) -sec-Butyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl) oxy) -2-fluoroacetate

Example 22-23.

Examples 22-23 were prepared in accordance with the procedure for Examples 1-2. To a solution of (2S, 5R) -6 (allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxamide (Intermediate 17, 0.202 g, 0, 85 mmol) in methanol (5 ml) at room temperature were added 1,3-dimethylbarbituric acid (0.266 g, 1.70 mmol) and tetrakis (triphenylphosphine) palladium (0) (0.098 g, 0.09 mmol). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated to give an orange film. The orange film was dissolved in DMF (5 ml) and potassium carbonate (0.353 g, 2.55 mmol) and (R) -s-butyl-2-bromo-2-fluoroacetate (Intermediate 23, 0.421 g, 1.98 mmol ). The reaction mixture was stirred for 3 h at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0-65% ethyl acetate / hexanes), (R) -sec-butyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetate (0.220 g, 78%) as an orange foam, mixture of diastereomers in a 1: 1 ratio. Separation of diastereomers was performed using reverse phase HPLC (Atlantis T3 19 mm x 150 mm, 30-50% acetonitrile in water, 20 ml / min, 15 min). The stereochemistry of the esters was assigned by hydrolysis of each of the esters to the corresponding acid and comparing the retention times in HPLC with the retention times of Examples 3 and 4.

Example 22 (First eluted peak) 90 mg, 32%, MS: 330 ES + (C ₁₄ H ₂₀ FN ₃ O ₅ ).

¹ H NMR (300 MHz, DMSO-d6) δ: 0.87 (t, 3H); 1.17 (d, 3H); 1.59 (m, 2H); 1.63 (s, 3H); 3.03 (m, 1H); 3.76 (d, 1H); 4.02 (m, 1H); 4.19 (s, 1H); 4.86 (m, 1H); 6.02 (m, 1H); 6.16-6.34 (m, 1H); 7.37 (bs, 1H); 7.82 (bs, 1H).

- 50 037916

Example 23 (Second eluting peak) 87 mg, 31%, MS: 330 ES + (C ₁₄ H ₂₀ FN3O5).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.88 (t, 3H); 1.24 (d, 3H); 1.61 (m, 2H); 1.63 (s, 3H); 3.08 (m, 1H); 3.76 (d, 1H); 3.94 (m, 1H); 4.21 (s, 1H); 4.89 (m, 1H); 6.04 (m, 1H); 6.03-6.22 (m, 1H); 7.38 (bs, 1H); 7.82 (bs, 1H).

Intermediate 24. Pentan-3-yl-2-bromo-2-fluoroacetate

Ethyl 2-bromo-2-fluoroacetate (0.639 ml, 5.41 mmol) was added to a mixture of dry pentan-3-ol (4.68 ml, 43.25 mmol) and hexane (20 ml). The resulting mixture was cooled to 0 ° C. KOtBu (0.091 g, 0.81 mmol) was added and the mixture was left stirring for 16 h at 25 ° C. Then the reaction was stopped by the addition of 1 N hydrochloric acid. HCl (30 ml), washed with water (50 ml) and brine (50 ml), dried with Na ₂ SO ₄ and concentrated. The crude material was purified by flash chromatography (20 g silica gel, 0100% Et ₂ O in hexane, 25 min) to give pentan-3-yl-2-bromo-2-fluoroacetate (0.754 g, 61.4%) as colorless oil.

1H NMR (300 MHz, chloroform-d) δ ppm 0.96 (td, J = 7.46, 2.08 Hz, 6 H) 1.60-1.76 (m, 4H) 4.93 (dt, J = 12.28, 6.33 Hz, 1H) 6.50 (s, 0.5H) 6.67 (s, 0.5H).

Example 24 (R) -Pentan-3-yl-2 - ((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6- yloxy) -2-fluoroacetate

Example 25. (S) -Pentan-3-yl-2 - ((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6- yloxy) -2-fluoroacetate

Example 24-25.

Examples 24-25 were prepared according to the procedure for Examples 1-2. To a solution of (2S, 5R) -6 (allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 17, 272 mg, 1, 15 mmol) in methanol (5 ml) at room temperature were added 1,3-dimethylbarbituric acid (358 mg, 2.29 mmol) and Pd (Ph ₃ P) ₄ (132 mg, 0.11 mmol). The reaction mixture was stirred at room temperature for 2 hours. The weight of the target product was monitored using

UPLC, no starting material was detected. The reaction mixture was concentrated to give an orange film. The orange film was dissolved in DMF (5 mL) and K ₂ CO ₃ (475 mg, 3.44 mmol) and pentan-3-yl-2-bromo-2-fluoroacetate (Intermediate 24, 751 mg, 3.31 mmol ). The reaction mixture was stirred overnight at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0-70% ethyl acetate / hexanes), pentan-3-yl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2 .1] oct3-en-6-yl) oxy) -2-fluoroacetate (320 mg, 81%) as a light yellow foam. It was confirmed by HPLC and NMR to be a 1: 1 mixture of diastereomers. Diastereomers were separated using reverse phase HPLC (Atlantis T3 4.6 mm x 50 mm 5 μm column, 30-50% ACN in water over 5 min, flow rate 1 ml / min).

Example 24.

125 mg, 32% HPLC. LCMS retention time, method 2min_Acid_CV10, acidic conditions: 0.86 min, 344 (M + H) ⁺ .

1H NMR (300 MHz, chloroform-d) δ ppm 0.92 (td, J = 7.46, 3.21 Hz, 6H) 1.61-1.75 (m, 4H) 1.93 (s, 3H ) 3.21-3.35 (m, 2H) 4.07 (dd, J = 4.91, 2.64 Hz, 1H) 4.36 (s, 1 H) 4.91 (quin, J = 6 , 18 Hz, 1H) 5.47 (br. S., 1H) 5.77 (s, 0.5H) 5.94 (s, 0.5H) 6.07-6.12 (m, 1H) 6 , 59 (br. S., 1H).

Example 25.

125 mg, 32% HPLC LCMS retention time, method 2min_Acid_CV10, acidic conditions: 0.91 min, 344 (M + H) +.

1H NMR (300 MHz, chloroform-d) 5 ppm 0.93 (t, J = 7.46 Hz, 3H) 0.95 (t, J = 7.46 Hz, 3H) 1.63-1.74 ( m, 4H) 1.93 (s, 3H) 3.20 - 3.38 (m, 2H) 4.02 (dd, J = 5.00, 2.55 Hz, 1H) 4.35 (s, 1 H) 4.92 (quin, J = 6.18 Hz, 1H) 5.56

- 51 037916 (br. S., 1H) 5.69 (s, 0.5H) 5.89 (s, 0.5H) 6.07-6.14 (m, 1H) 6.64 (br. S ., 1H)

Intermediate 25. N - [(3R, 6S) -6 - [[(tert-butyldimethylsilyl) oxy] methyl] -5-methyl1,2,3,6-tetrahydropyridin-3-yl] -2-nitro-N- (prop-2-en-1-yloxy) benzene-1-sulfonamide

ITBSO ^^ 'p ^ l

Ns

A solution of tert-butyl- (3R, 6 S) -6- [[(tert-butyldimethylsilyl) -oxy] methyl] -5-methyl-3 - [ N (prop-2-en-1-yloxy) (2-nitrobenzene) sulfonamido] -1,2,3,6-tetrahydropyridine-1-carboxylate (Intermediate 11, 13.6 g, 22.75 mmol, 1 eq .) in dichloromethane (100 ml). _{Then ZnBr 2} (10.2 g, 45.29 mmol, 2 eq.) Was added in several batches. The resulting solution was stirred overnight at room temperature. The resulting solution was diluted with 500 ml of dichloromethane. The resulting mixture was washed with 2x200 ml sodium bicarbonate (aq) and 2x200 ml NH4Cl (aq). The mixture was dried over anhydrous sodium sulfate and concentrated in vacuo. This gave 12 g (crude product) of the title compound as a yellow oil. MS: 498 ES + (C ₂₂ H ₃₅ N ₃ O ₆ SSi).

Intermediate 26. (3R, 6S) -6 - [[(tert-Butyldimethylsilyl) oxy] methyl] -5-methyl-N (prop-2-en-1 -yloxy) -1,2,3,6-tetrahydropyridine -3-amine

TBS (Z-p4 hn ^ _n .o ^ Η

A solution of N - [(3R, 6S) -6 - [[(tert-butyldimethylsilyl) oxy] methyl] -5-methyl-1,2,3, 6-tetrahydropyridin-3-yl] -2-nitro-N- (prop-2-en-1 -yloxy) benzene-1-sulfonamide (intermediate 25, 12 g, 24.11 mmol, 1 eq.) In N , N-dimethylformamide (100 ml), 2-sulfanylacetic acid (4.4 g, 47.77 mmol, 2 eq.). Then LiOH (5.8 g, 242.17 mmol, 10 eq.) Was added in portions. The resulting solution was stirred for 2 h at room temperature, then diluted with 500 ml of water and extracted with 5x200 ml of ethyl acetate, and the organic layers were combined. The organic mixture was washed with 3x200 ml brine and 2x200 ml sodium bicarbonate (aq). The mixture was dried over anhydrous sodium sulfate and concentrated in vacuo. This gave 8.4 g (crude product) of the title compound as a yellow oil. MS: 313 ES + (C ₁₆ H ₃₂ N ₂ O ₂ Si).

Intermediate 27. (2S, 5R) -2 - [[(tert-Butyldimethylsilyl) oxy] methyl] -3-methyl-6- (prop2-en-1 -yloxy) -1,6-diazabicyclo [3.2.1] oct-3-en-7-one

TBSO ^{/ Z} '''^%

J— ^N o

A solution of (3R, 6S) -6 - [[(tert-butyldimethylsilyl) oxy] methyl] -5-methyl-N- (prop-2en-1 -yloxy) -1,2,3,6-tetrahydropyridin-3-amine (Intermediate 26, 8.4 g, 26.88 mmol, 1 eq.) in acetonitrile (1.6 L) and N, N-diisopropylethylamine (14.2 g, 109.87 mmol, 4 eq.). Then, a solution of ditrichloromethyl carbonate (2.9 g, 9.77 mmol, 0.4 eq.) In acetonitrile (100 ml) was added dropwise with stirring at -15 ° C for 3 hours. The resulting solution was stirred overnight at room temperature ... The resulting mixture was concentrated in vacuo. The crude product was diluted with 500 ml of ethyl acetate. The resulting mixture was washed with 2x400 ml NH ₄ Cl (aq) and 2x400 ml brine, then concentrated in vacuo. The residue was eluted on a silica gel column with ethyl acetate / petroleum ether (1:10). This gave 3.9 g (43%) of the title compound as a yellow oil. MS: 339 ES + (C _n H _{3 (} #O ^).

Intermediate 28. (2S, 5R) -2- (Hydroxymethyl) -3-methyl-6- (prop-2-en-1-yloxy) -1.6 diazabicyclo [3.2.1] oct-3-en-7- he

Tetrahydrofuran (30 ml) and (2S, 5R) -2 - [[(tert-butyldimethylsilyl) oxy] methyl] -3-methyl-6- (prop-2-en-1 -yloxy) -1 , 6-diazabicyclo [3.2.1] oct-3-en-7-one (Intermediate 27, 3.2 g, 9.45 mmol, 1 eq.) And the solution was cooled to 0 ° C, then TBAF was added dropwise (14.2 ml 1N in THF, 1.5 eq.). The reaction mixture was stirred for 1 h at 0 ° C in an ice / water bath. The resulting mixture was concentrated in vacuo. The residue was eluted on a silica gel column with ethyl acetate / petroleum ether (1: 5-1: 2). This gave 1.6 g (75%) of the title compound as a light yellow solid. MS: 225 ES + (CnH ₁₆ N ₂ O ₃ ).

- 52 037916

Ή NMR (300 MHz, CDCI3) δ 1.63 (3H, d), 3.20 (2H, d), 3.62-3.84 (2H, m), 3.85-3.90 (2H, m), 4.35-4.48 (2H, m), 5.28-5.39 (2H, m), 5.95-6.08 (2H, m).

Intermediate 29. (2S, 5R) -3-Methyl-7-oxo-6- (prop-2-en-1-yloxy) -1,6-diazabicyclo

[3.2.1] oct-3-ene-2-carboxylic acid

О I but ^А D

J - Ν

O

To a solution of H ₅ IO ₆ (12.93 g, 56.71 mmol) in wet CH ₃ CN (150 ml, 0.75 vol.% H ₂ O) at room temperature. pace. added CrO ₃ (128 mg, 1.28 mmol). The mixture was stirred until complete dissolution. Wet acetonitrile- (35 ml) and (2S, 5R) -2- (hydroxymethyl) -3-methyl-6 (prop-2-en-1-yloxy) -1,6-diazabicyclo [ 3.2.1] oct-3-en-7-one (Intermediate 28, 740 mg, 3.30 mmol, 1 eq.), And cooled to 0 ° C. Then, the above oxidizing agent solution (35 ml, 3 eq.) Was added dropwise over 30 min at 0 ° C. The resulting solution was stirred for 1 hour at 0 ° C. in an ice / water bath. Then the reaction mixture was diluted with 200 ml of chloroform and 50 ml of citric acid solution (25%). The organic layer was separated and then washed with 3x50 ml brine, dried over anhydrous sodium sulfate and concentrated in vacuo. This gave 0.70 g of the crude title compound as a yellow oil. MS: 239 ES + (CnH ₁₆ N ₂ O ₄ ).

Intermediate 30. (2S, 5R) -N'-Acetyl-6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carbohydrazide

To a solution of (2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 29, 195.7 mg, 0.82 mmol) in DCM (10 ml), 1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride (236 mg, 1.23 mmol), 1-hydroxybenzotriazole hydrate (219 mg, 1.23 mmol) and monoacetylhydrazine (101 mg, 1.23 mmol). The mixture was cooled to 0 ° C and DIEA (0.715 ml, 4.11 mmol) was added. The reaction mixture was allowed to warm up to room temperature. pace. and stirred at room temperature overnight, then diluted with ethyl acetate and washed twice with a 1: 1 mixture of brine and water. The aqueous wash solutions contained some product and were stripped twice with ethyl acetate and once with about 5% methanol in dichloromethane. The combined organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-5% methanol / dichloromethane) afforded the title compound as a white foam (45.5 mg, 19%, ca.50% purity). MS: 295 ES + (C13H18N4O4).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.62 (s, 3H); 1.86 (m, 3H); 2.88 (m, 1H); 3.40 (m, 1H); 3.81 (m, 1H); 3.95 (m, 1H); 4.38 (m, 2H); 5.32 (m, 2H); 5.94 (m, 1H); 6.11 (m, 1H); 9.86 (s, 1H); 10.24 (bs, 1H).

Example 26. Ethyl-2 - (((2S, 5R) -2- (2-acetylhydrazinecarbonyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl ) hydroxy) -2-fluoroacetate

O

To a solution of (2S, 5R) -N'-acetyl-6- (allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2carbohydrazide (Intermediate 30, 22 , 75 mg, 0.08 mmol,) in methanol (3 ml), 1,3-dimethylbarbituric acid (48.3 mg, 0.31 mmol) and tetrakis (triphenylphosphine) palladium (0) (17.87 mg, 0.02 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated to give an orange film. The orange film was dissolved in DMF (3 ml). Potassium chloride (64.1 mg, 0.46 mmol) and ethyl 2-bromo-2-fluoroacetate (0.055 ml, 0.46 mmol) were added. The reaction mixture was stirred for about 5 hours at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-65% ethyl acetate / hexanes) afforded the title compound as an orange foam (25.9 mg, 94%). The two diastereomers were present in a 1: 1 ratio. The material was dissolved in ethyl acetate and washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. Elute again on a silica gel column (0-30% acetone / dichloromethane) to give the pure title compound

- 53 037916 (16.6 mg, 60%). MS: 359 ES + (C ₁₄ H ₁₉ FN4O ₆ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.24 (m, 3H); 1.65 (s, 3H); 1.86 (m, 3H); 3.09 (m, 1H); 3.89 (d, 1H); 4.01 (m, 1H); 4.25 (m, 3H); 6.09 (m, 1H); 6.20 (m, 1H); 9.90 (s, 1H); 10.30 (d, 1H).

Example 27.2 - (((2S, 5R) -2- (2-Acetylhydrazinecarbonyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-en-6-yl) oxy) - 2-fluoroacetic acid lithium salt

To a solution of ethyl-2 - (((2S, 5R) -2- (2-acetylhydrazinecarbonyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) hydroxy) -2-fluoroacetate (example 26, 16.6 mg, 0.05 mmol) in THF (1 ml) and water (0.33 ml) at 0 ° C was added lithium hydroxide (1M) (0.05 ml, 0.05 mmol) and stirred for 10 min at 0 ° C. Added another 0.2 eq. lithium hydroxide. After 30 min, another 0.2 eq. lithium hydroxide. The reaction mixture was stirred for an additional 30 minutes. HCl (0.5M) (0.046 ml, 0.02 mmol) was added to adjust the pH to about 4-5. The reaction mixture was extracted twice with ethyl acetate. The aqueous layer was frozen and lyophilized. This gave 15 mg of the title compound as an orange solid. The two diastereomers were present in a 1: 1 ratio. MS: 331 ES + (C ₁₂ H ₁₅ FN ₄ O ₆ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.65 (s, 3H); 1.87 (m, 3H); 3.08 (m, 1H); 3.84 (m, 1H); 4.03 (m, 1H); 4.23 (s, 1H); 5.23 (m, 1H); 6.10 (m, 1H); 9.88 (s, 1H); 10.26 (bs, 1H).

Intermediate 31. tert-Butyl-4 - ((2S, 5R) -6- (allyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-encarboxamido) benzyl carbamate

To a solution of (2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxylic acid (Intermediate 29, 130.5 mg, 0.55 mmol) in DMF (5 ml), tert-butyl 4-aminobenzylcarbamate (146 mg, 0.66 mmol), Hunig's base (0.287 ml, 1.64 mmol) and cyclic anhydride 1- propane phosphonic acid (50 wt% in DMF) (0.326 ml, 1.10 mmol). The mixture was stirred at room temperature for 1 h. Water (10 ml) and 10% MeOH in DCM (20 ml) were added. The organic layer was separated and concentrated to give the crude product. Purification by flash chromatography (20 g silica gel, 0% -10% MeOH in DCM, 20 min) afforded the title compound (178 mg, 73.4% yield, ca. 50% purity) as a yellow oil. MS: 443 ES + (C ₂₃ H ₃₀ N ₄ O ₅ ).

1H NMR (300 MHz, chloroform-d) δ ppm 1.47 (m, 12H) 3.10-3.23 (m, 1H) 3.25-3.39 (m, 1H) 4.34-4, 54 (m, 3H) 4.36-4.49 (m, 3H) 5.25-5.45 (m, 2H) 6.03 (d, J = 6.61 Hz, 1H) 6.09-6 , 17 (m, 1H) 7.24-7.29 (m, 2H) 7.427.62 (m, 2H) 8.64 (s, 1H).

Intermediate 32. (R) -ethyl-2 - ((2S, 5R) -2- (4 - ((tert-butoxycarbonylamino) methyl) phenylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2 .1] oct-3-en-6-yloxy) -2-fluoroacetate

To a solution of tert-butyl 4 - ((2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2carboxamido) benzylcarbamate ( Intermediate 31, 178 mg, 0.40 mmol) in methanol (10 ml) was added Pd (Ph ₃ P) ₄ (465 mg, 0.40 mmol) at room temperature. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated to give the crude intermediate tert-butyl-4 - ((2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [ 3.2.1] oct-3-encarboxamido) benzyl carbamate in the form of an orange film. The crude material was dissolved in DMF (1 ml). K ₂ CO ₃ (167 mg, 1.21 mmol) and ethyl 2-bromo-2-fluoroacetate (0.052 ml, 0.44 mmol) were added. The reaction mixture was stirred overnight at room temperature. EtOAC (20 ml) was added and the reaction mixture was washed with water (10 ml). The organic layer was concentrated to give the crude product, which was purified by flash chromatography (12 g silica gel, 0-100% EtOAc in hexane, 20 min; then 5% MeOH in DCM, 10 min) to give the indicated 54 037916 the title compound (11 mg, 5.4% yield) as an orange solid. MS: 507 ES + (C24H31FN4O7).

Ή NMR (300 MHz, chloroform-d) δ ppm 1.36 (t, 3H) 1.48 (s, 9H) 2.00 (s, 3H) 3.21-3.27 (m, 1H) 3, 36 (d,

J = 1.70 Hz, 1H) 4.08 (dd, J = 5.00, 2.74 Hz, 1H) 4.24-4.42 (m, 5H) 4.47 (s, 1H) 5, 78-5.95 (m, 1H) 6.12-6.17 (m,

1H) 7.38-7.55 (m, 4H) 8.47 (s, 1H).

Intermediate 33. (R) -2 - ((2S, 5R) -2- (4 - ((tert-Butoxycarbonylamino) methyl) phenylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1 ] oct-3-en-6-yloxy) -2-fluoroacetic acid lithium salt

he

To a solution of (2R) -ethyl-2 - (((2S, 5R) -2 - ((4 - (((tert-butoxycarbonyl) amino) methyl) phenyl) carbamoyl) -3methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetate (Intermediate 32, 11 mg, 0.02 mmol) in THF (1 ml) and water (0.5 ml) at 0 ° C lithium hydroxide (1M) (0.02 ml, 0.02 mmol) was added. The mixture was stirred for 5 min at 0 ° C and DCM (10 ml) was added. Hydrochloric acid (0.5N) was added carefully to adjust the pH to about 5-6. The organic layer was separated and concentrated to give the title compound (8 mg, 77%) as a yellow solid. HPLC, acidic conditions, retention time: 0.80 min, 479 (M + H) +.

Ή NMR (300 MHz, chloroform-d) δ ppm 1.47 (s, 9H) 1.98 (s, 3H) 3.19-3.44 (m, 1H) 4.03-4.55 (m, 6H) 5.68-6.05 (m, 1H) 7.44-7.56 (m, 4H) 8.49 (br. S., 1H).

Example 28. (R) -2 - ((2S, 5R) -2- (4- (Aminomethyl) (phenylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3- en-6-yloxy) -2-fluoroacetic acid TFA salt

he

To a solution of lithium salt (2R) -2 - (((2S, 5R) -2 - ((4 - (((tert-butoxycarbonyl) amino) methyl) phenyl) carbamoyl) -3-methyl-7-oxo-1, 6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetic acid (Intermediate 33, 8 mg, 0.02 mmol) in DCM (2 ml) at 0 ° C was added TFA (0.128 ml, 1.67 mmol). The reaction mixture was stirred at 0 ° C for 6 h and the solvent was removed. Toluene (2x1 ml) was added and concentrated to remove excess TFA. Added Et ₂ O (2 ml) was added and the formed precipitate was filtered off, washed with Et ₂ O and dried to give the title compound (8 mg, 97%) as a TFA salt. MS: 379 ES + (C ₁₇ H ₁₉ FN ₄ O ₅ ).

Ή NMR (300 MHz, D2O) δ ppm 1.75 (s, 3H) 3.23-3.61 (m, 2H) 4.12-4.25 (m, 3H) 4.51-4.60 ( m, 1H) 5.53-5.98 (m, 1H) 6.29 (br. s., 1H) 7.45-7.57 (m, 4H).

Intermediate 34. (2S, 5R) -6- (Allyloxy) -3-methyl-7-oxo-N- (pyrazin-2-ylmethyl) -1.6 diazabicyclo [3.2.1] oct-3-ene-2- carboxamide

To a solution of (2S, 5R) -6- (αlyloxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 29, 199 mg, 0.84 mmol) in DMF (5 ml) at 0 ° С, pyrazin-2-ylmethanamine (91 mg, 0.84 mmol), O- (7-azabenzotriazol-1-yl) -N, N, N ', N'-tetramethyluronium hexafluorophosphate (635 mg, 1.67 mmol) and DIEA (0.582 ml, 3.34 mmol). The reaction mixture was stirred for 30 min at room temperature, then diluted with ethyl acetate and washed once with saturated sodium bicarbonate solution and three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-2.5% methanol / dichloromethane) gave the title compound (147 mg, 53.5%) as an orange oil. MS: 330 ES + (C ₁₆ H ₁₉ N ₅ O ₃ ).

NMR (300 MHz, DMSO-d ₆ ) δ: 1.60 (s, 3H); 3.14 (m, 2H); 3.64 (m, 1H); 3.94 (m, 1H); 4.35 (m, 2H); 4.46 (m, 2H); 5.29 (m, 2H); 5.96 (m, 1H); 6.07 (m, 1H); 8.56 (m, 3H); 9.05 (m, 1H).

Example 29. Ethyl-2-fluoro-2 - (((2S, 5R) -3-methyl-7-oxo-2 - ((pyrazin-2-ylmethyl) carbamoyl) -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) acetate

- 55 037916

To a solution of (2S, 5R) -6- (allyloxy) -3-methyl-7-oxo-N- (pyrazin-2-ylmethyl) -1,6-diazabicyclo [3.2.1] oct-3-en-2- carboxamide (Intermediate 34, 153.8 mg, 0.47 mmol) in methanol (3 ml), 1,3-dimethylbarbituric acid (146 mg, 0.93 mmol) and tetrakis (triphenylphosphine) palladium (0) were added at room temperature (54.0 mg, 0.05 mmol). The reaction mixture was stirred at room temperature for 2 hours. Tetrakis (triphenylphosphine) palladium (0) (54.0 mg, 0.05 mmol) was added again, as well as 2 ml of methanol, and the mixture was stirred at room temperature for another 2 hours. The reaction mixture was concentrated to give an orange oil. The oil was dissolved in DMF (3 ml) and potassium carbonate (194 mg, 1.40 mmol) and ethyl 2-bromo-2-fluoroacetate (0.166 ml, 1.40 mmol) were added. The reaction mixture was stirred overnight at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0% -100% ethyl acetate / hexanes), the title compound (95 mg, 52%) was obtained as a light orange foam. The compound was a 1: 1 mixture of diastereomers. MS: 394 ES + (C17H20FN5O5).

1H NMR (300 MHz, DMSO-a6) δ: 1.22 (m, 3H); 1.63 (s, 3H); 2.71 (m, 1H); 3.10 (m, 1H); 3.98 (d, 1H); 4.02 (m, 1H); 4.26 (m, 2H); 4.47 (m, 2H); 6.05 (m, 1H); 6.20 (m, 1H); 8.56 (m, 3H); 9.10 (m, 1H).

Example 30. 2-Fluoro-2 - (((2S, 5R) -3-methyl-7-oxo-2 - ((pyrazin-2-ylmethyl) carbamoyl) -1,6-diazabicyclo [3.2.1] oct- 3-en-6-yl) oxy) acetic acid lithium salt

To a solution of ethyl 2-fluoro-2 - (((2S, 5R) -3-methyl-7-oxo-2 - ((pyrazin-2-ylmethyl) carbamoyl) -1.6 diazabicyclo [3.2.1] oct-3 -en-6-yl) oxy) acetate (Example 29, 95.3 mg, 0.24 mmol) in THF (2 ml) and water (0.66 ml) at 0 ° C, lithium hydroxide (1M) (0.254 ml, 0.25 mmol) and stirred for 25 min at 0 ° C. Added another 0.1 eq. lithium hydroxide. After 15 minutes, hydrochloric acid (0.5N) (0.242 mL, 0.12 mmol) was added to adjust the pH to about 5-6. The reaction mixture was frozen and lyophilized. 90 mg of a light yellow solid was purified by reverse phase HPLC (YMC Carotenoid C30, 21.2 mm x 150 mm, 4 μm conjugated with Synergi Polar RP, 21.2 mm x 100 mm, 4 μm, 0% - 30% acetonitrile in water, 20 ml / min, 15 min). This gave 23.6 mg (27%) of the title compound as a white solid. The compound was a 1: 1 mixture of diastereomers. MS: 366 ES + (C ₁₅ H ₁₆ FN ₅ O5).

1H NMR (300 MHz, DMSO-d6) δ: 1.63 (s, 3H); 3.09 (m, 1H); 3.68 (m, 1H); 4.02 (m, 1H); 4.29 (s, 1H); 4.48 (d, 2H); 5.22 (m, 1H); 6.05 (m, 1H); 8.57 (m, 3H); 9.07 (m, 1H).

Intermediate 35. (S) -tert-Butyl-1- (tert-butyldimethylsilyloxy) but-3-en-2-yl (3methyl-2-oxobut-3-enyl) carbamate

TBSO ^^ Y ^

To -N boc

O

To a solution of (S) -tert-butyl-1 - (tert-butyldimethylsilyloxy) but-3-en-2-yl (2- (methoxy (methyl) amino) 2-oxoethyl) carbamate (Intermediate 5, 30.79 g , 76.48 mmol) in THF (200 ml) at 0 ° С was added prop-1-en-2-ylmagnesium bromide (0.5 M in THF) (300 ml, 149.90 mmol) and stirred at 0 ° С in for 1 h. The reaction was stopped by adding 200 ml of 10% citric acid, additionally diluted with 100 ml of water and extracted with simple ether. The organics were concentrated and the resulting oil was dissolved in ether and washed with water and brine. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0% -20% ethyl acetate / hexanes) gave the title product (26.2 g, 89%) as a colorless oil. MS: 384 ES + (C20H37NO4S1).

1H NMR (300 MHz, DMSO-d6) δ: 0.02 (d, 6H); 0.83 (s, 9H); 1.27-1.38 (m, 9H); 1.80 (m, 3H); 3.71 (m, 2H); 4.34 (m, 2H); 4.61 (m, 1H); 5.17 (m, 2H); 5.77 (m, 1H); 5.85 (m, 1H); 6.03 (m, 1H).

Intermediate 36. (S) -tert-Butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -4-methyl-5oxo-5,6-dihydropyridine-1 (2H) -carboxylate

- 56 037916 tbso γ boc О Solution of (S) -tert-butyl-1- (tert-butyldimethylsilyloxy) but-3-en-2-yl (3-methyl-2-oxobut-3-enyl) carbamate (Intermediate 35 , 26.18 g, 68.25 mmol) in toluene (600 mL) was purged with nitrogen for 15 min. Then (1,3-bis- (2,4,6-trimethylphenyl) -2-imidazolidinylidene) dichloro (oisopropoxyphenylmethylene) ruthenium (0.987 g, 1.57 mmol) was added. The reaction mixture was at 65 ° C for 1.5 h. The reaction mixture was concentrated on silica gel. Chromatography on silica gel (0% -15% ethyl acetate / hexanes) gave the title product (21.18 g, 87%) as a colorless oil. MS: 356 ES + (C18H33NO ₄ Si).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.01 (d, 6H); 0.81 (s, 9H); 1.42 (s, 9H); 1.75 (m, 3H); 3.74-3.89 (m, 3H); 4.04-4.32 (m, 1H); 4.67 (m, 1H); 6.88 (m, 1H).

Intermediate 37. (2S, 5S) -tert-Butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -5hydroxy-4-methyl-5,6-dihydropyridine-1 (2H) -carboxylate boc OH

To a solution of cerium (III) chloride (14.68 g, 59.57 mmol) and (S) -tert-butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -4-methyl-5-oxo-5,6- dihydropyridine-1 (2H) -carboxylate (Intermediate 36, 21.18 g, 59.57 mmol) in methanol (300 ml), sodium borohydride (2.254 g, 59.57 mmol) was added in portions at 0 ° C. After 15 minutes, the reaction mixture was diluted with saturated ammonium chloride solution (100 ml) and water (100 ml), then extracted twice with diethyl ether. The organic extracts were washed with brine, dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-20% ethyl acetate / hexanes) gave the title product (19.45 g, 91%) as a colorless oil. MS: 358 ES + (C ₁₈ H ₃₅ NO ₄ Si).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.02 (s, 6H); 0.86 (s, 9H); 1.39 (s, 9H); 1.69 (m, 3H); 2.63-2.72 (m, 1H); 3.59 (m, 2H); 3.82 (m, 1H); 4.03 (m, 1H); 4.21 (m, 1H); 5.04 (d, 1H); 5.38 (m, 1H).

Intermediate 38. N- (Allyloxy) -2-nitrobenzenesulfonamide

To a stirred solution of O-allylhydroxylamine hydrochloride (147.05 g, 1341.59 mmol) in DCM (2.5 L) at 0 ° C was added pyridine (318 ml, 3948 mmol), followed by 2-nitrobenzene-1sulfonyl chloride ( 250 g, 1128.05 mmol) in solid form. The reaction mixture was then stirred at the same temperature for 1 hour. The completion of the reaction was monitored by TLC. The reaction was stopped by the addition of 1.5 N. HCl (1 L). The organic layer was separated, washed with water (250 ml), brine (250 ml), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to give a residue. The crude material was purified by crystallization using a mixture of EtOAc and petroleum ether (1: 3) (800 ml) to obtain 202 g of the title compound as a light brown solid.

The mother liquor was concentrated and purified by column chromatography on silica gel (60-120 mesh) using a mixture of EtOAc and petroleum ether (7: 3) to give an additional 19.1 g of the title compound as a yellow solid. The overall yield was 76%. UPLC: 257 (M-1) for C ₉ H ₁₀ N ₂ O ₅ S.

Ή NMR (400 MHz, DMSO-d6): δ 4.36-4.38 (m, 2H), 5.22-5.32 (m, 2H), 5.84-5.91 (m, 1H) , 7.92-7.96 (m, 2H), 8.02-8.05 (m, 2H), 11.07 (s, 1H).

Intermediate 39. (2S, 5R) -tert-Butyl-5- (N- (allyloxy) -2-nitrophenylsulfonamido) 2 - ((tert-butyldimethylsilyloxy) methyl) -4-methyl-5,6-dihydropyridine-1 ( 2H) -carboxylate

To a solution of (2S, 5S) -tert-butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -5-hydroxy-4-methyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 37, 19.45 g , 54.40 mmol) in toluene (300 ml) at room temperature was added triphenylphosphine (17.06 g, 65.28 mmol), N- (allyloxy) 2-nitrobenzenesulfonamide (Intermediate 38, 14.05 g, 54.40 mmol) and diisopropyl azodicarboxylate (12.85 ml, 65.28 mmol). After 2 hours, the reaction mixture was concentrated on silica gel and purified. Chromatography on silica gel (0% -50% ethyl acetate / hexanes) gave the title product (25.2 g, 78%) as a yellow oil. MS: 598 ES + (C ₂₇ H ₄₃ N ₃ O ₈ SSi).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 0.00 (s, 6H); 0.83 (s, 9H); 1.31 (m, 9H); 1.34 (m, 3H); 3.10-3.25 (m, 1H); 3.59 (m, 2H); 3.99-4.41 (m, 5H); 5.17 (m, 2H); 5.72 (m, 2H); 7.93-8.16 (m, 4H).

Intermediate 40. (2S, 5R) -tert-Butyl-5- (N- (allyloxy) -2-nitrophenylsulfonamido) - 57 037916

2- (hydroxymethyl) -4-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

A XL boc N

Ns

To a solution of (2S, 5R) -tert-butyl-5- (N- (allyloxy) -2-nitrophenylsulfonamido) -2 - ((tert-butyldimethylsilyloxy) methyl) -4-methyl-5,6-dihydropyridine-1 (2H ) -carboxylate (Intermediate 39, 1 g, 1.67 mmol) in THF (11 ml) at 0 ° C was added tetrabutylammonium fluoride (1M in THF) (2.175 ml, 2.17 mmol). After 90 minutes, the reaction mixture was concentrated on silica gel. Chromatography on silica gel (0% -70% ethyl acetate / hexanes) gave the title product (0.732 g, 90%) as a brownish-beige foam. MS: 484 ES + (C ₂₁ H ₂₉ N ₃ O ₈ S).

1 H NMR (300 MHz, DMSO-d ₆ ) δ: 1.31 (m, 9H); 1.35 (m, 3H); 3.20 (m, 1H); 3.41 (m, 2H); 3.96-4.37 (m, 5H); 4.76 (m, 1H); 5.19 (m, 2H); 5.66-5.84 (m, 2H); 7.94-8.18 (m, 4H).

Intermediate 41. (2S, 5R) -5- (N- (Allyloxy) -2-nitrophenylsulfonamido) -1- (tert-butoxycarbonyl) -4-methyl-1,2,5,6-tetrahydropyridine-2-carboxylic acid o

L _ / no ru boX ^N

Ns

To a solution of iodic acid (6 g, 31.26 mmol) in wet acetonitrile (60 ml) (0.75 vol% water) at room temperature was added chromium (VI) oxide (10 mg, 0.10 mmol). The mixture was stirred until complete dissolution. To a solution of (2S, 5R) -tert-butyl 5- (N- (αlyloxy) -2-nitrophenylsulfonamido) -2 (hydroxymethyl) -4-methyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 40.5 g, 10.34 mmol) in wet acetonitrile (60 ml) (0.75 vol.%) At 0 ° C was added dropwise the previously prepared iodic acid / chromium oxide solution (60 ml, 3 eq.). After 30 min, the reaction mixture was diluted with ether and washed with 10% citric acid, saturated sodium bicarbonate solution and brine. The organics were dried over magnesium sulfate, filtered and concentrated to give an orange foam (4.16 g, 81%). MS: 498 ES + (C ₂₁ H ₂₇ N ₃ O ₉ S).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.26 (m, 9H); 1.31 (m, 3H); 3.02-3.25 (m, 1H); 3.90 (m, 1H); 4.17 (m, 3H); 4.65-4.77 (m, 1H); 5.12-5.21 (m, 2H); 5.68 (m, 1H); 5.88 (m, 1H); 7.92-8.17 (m, 4H).

Intermediate 42. (2S, 5R) -tert-Butyl-5- (N- (αlyloxy) -2-nitrophenylsulfonamido) 2-carbamoyl-4-methyl-5,6-dihydropyridine-1 (2H) -carboxylate o

A h ₂ n py .CL boc N

Ns

To a solution of (2S, 5R) -5- (N- (αlyloxy) -2-nitrophenylsulfonamido) -1- (tert-butoxycarbonyl) -4methyl-1,2,5,6-tetrahydropyridine-2-carboxylic acid (Intermediate 41 , 4.16 g, 8.36 mmol) in DMF (35 ml), ammonium chloride (0.895 g, 16.72 mmol), HATU (4.77 g, 12.54 mmol) and DIEA (5, 84 ml, 33.45 mmol). After 15 minutes, the reaction mixture was diluted with ethyl acetate, washed with saturated sodium bicarbonate solution and twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0% -80% ethyl acetate / hexanes) twice, the title product (2.16 g, 52%) was obtained as a yellow foam. MS: 497 ES + (C ₂₁ H ₂₈ N ₄ O ₈ S).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.26 (m, 9H); 1.37 (m, 3H); 3.12-3.35 (m, 1H); 3.80 (m, 1H); 4.18 (m, 3H); 4.64-4.79 (m, 1H); 5.13-5.22 (m, 2H); 5.68 (m, 1H); 5.88 (m, 1H); 7.04 (m, 1H); 7.45 (bs, 1H); 7.90-8.18 (m, 4H).

Intermediate 43. (2S, 5R) -5- (N- (Allyloxy) -2-nitrophenylsulfonamido) -4-methyl1,2,5,6-tetrahydropyridine-2-carboxamide

To a solution of (2S, 5R) -tert-butyl 5- (N- (αlyloxy) -2-nitrophenylsulfonamido) -2-carbamoyl-4methyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 42, 2 , 16 g, 4.35 mmol) in DCM (20 ml) was added zinc bromide (0.700 ml, 13.05 mmol) at room temperature. After stirring overnight at room temperature, the reaction mixture was diluted with dichloromethane and washed with saturated sodium bicarbonate solution and brine. The organics were dried over magnesium sulfate, filtered and concentrated to give the title product (1.450 g, 84%) as a yellow foam. MS: 397 ES + (C ₁₆ H ₂₀ N ₄ O ₆ S).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.65 (m, 3H); 2.71 (m, 3H); 3.76 (m, 1H); 3.95 (m, 1H); 4.18-4.42 (m, 2H); 5.23 (m, 2H); 5.82 (m, 1H); 6.02 (m, 1H); 7.05 (bs, 1H); 7.30 (bs, 1H); 7.93-8.18 (m, 4H).

- 58 037916

Intermediate 44. (2S, 5R) -5- (allyloxyamino) -4-methyl-1,2,5,6-tetrahydropyridine-2carboxamide and (2R, 5R) -5- (allyloxyamino) -4-methyl-1, 2,5,6-tetrahydropyridine-2-carboxamide

To a solution of (2S, 5R) -5- (N- (allyloxy) -2-nitrophenylsulfonamido) -4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide (Intermediate 43, 1.4 g, 3 , 53 mmol) and cesium carbonate (9.21 g, 28.25 mmol) in THF (100 ml) at room temperature was added PS-thiophenol (3- (3-mercaptophenyl) propanamidomethylpolystyrene) (1.55 mmol / g) ( 9.12 g, 14.13 mmol). After stirring overnight at room temperature, the reaction mixture was filtered through a sintered glass funnel and the resin was washed twice with DCM. The filtrate was concentrated to give a yellow oil. After chromatography on silica gel (0-5% methanol / dichloromethane), 3: 1 mixtures of trans and cis isomers (0.473 g, 63.4%) were obtained as a light yellow oil. The mixture was used further without separation. MS: 212 ES + (C ₁₀ H ₁₇ N ₃ O ₂ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.73 (m, 3H); 2.63 (m, 1H); 2.97 (m, 1H); 3.01 (m, 1H); 3.60 (m, 1H); 4.12 (m, 2H); 5.11-5.26 (m, 2H); 5.92 (m, 1H); 6.45 (m, 1H); 7.00 (m, 1H); 7.33 (bs, 1H).

Intermediate 45. (2S, 5R) -6- (Allyloxy) -4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-ene-2-carboxamide

To a solution of (2S, 5R) -5- (allyloxyamino) -4-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide and (2R, 5R) -5- (allyloxyamino) -4-methyl-1, 2,5,6-tetrahydropyridine-2-carboxamide (Intermediate 44, 0.429 g, 2.03 mmol) and DIEA (1.415 ml, 8.12 mmol) in acetonitrile (170 ml) triphosgene (0.241 g , 0.81 mmol) as a solution in acetonitrile (1.5 ml) at a rate of 0.1 ml / min. After the addition was complete, the reaction mixture was warmed to room temperature and stirred for two days. The reaction mixture was diluted with ethyl acetate, washed with saturated sodium bicarbonate solution and brine, dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0% -20% ethyl acetate / hexanes) gave the product (0.312 g, 64.8%) as a light yellow oil. MS: 238 ES + (C ₁₁ H ₁₅ N ₃ O ₃ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.79 (m, 3H); 3.19 (m, 2H); 3.81 (m, 1H); 4.12 (m, 1H); 4.36 (m, 2H); 5.24-5.45 (m, 3H); 5.89-6.00 (m, 1H); 7.28 (bs, 1H); 7.49 (bs, 1H).

Example 31. (2R) -Ethyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy ) -2-fluoroacetate

Example 32. (2S) -Ethyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy ) -2-fluoroacetate

Examples 31-32.

To a solution of (2S, 5R) -6- (allyloxy) -4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 45, 0.2972 g , 1.25 mmol) in methanol (6 ml) at room temperature were added 1,3-dimethylbarbituric acid (0.391 g, 2.51 mmol) and tetrakis (triphenylphosphine) palladium (0) (0.145 g, 0.13 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated to give an orange film. The orange film was dissolved in DMF (6 ml). Potassium carbonate (0.519 g, 3.76 mmol) and ethyl bromofluoroacetate (0.592 ml, 5.01 mmol) were added. The reaction mixture was stirred overnight at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0-65% ethyl acetate /

- 59 037916 hexanes) obtained a mixture of diastereomers in the ratio 1: 1, 372 mg, 99%. Separation of diastereomers was performed using reverse phase HPLC (Atlantis T3, 19 mm x 150 mm, 5 μm, 20-40% acetonitrile in water, 20 ml / min, 15 min). After lyophilization, both diastereomers were obtained as white solids. The following products were obtained.

Example 31 (First eluting diastereomer): 108.8 mg, 29% MS: 302 ES + (C ₁₂ H ₁₆ FN ₃ O ₅ ).

¹ H NMR (300 MHz, DMSO-d6) δ: 1.23 (t, 3H); 1.82 (m, 3H); 3.19 (m, 1H); 3.29 (m, 1H); 3.96 (m, 1H); 4.23 (m, 1H); 4.24 (q, 2H); 5.52 (m, 1H); 6.28 (m, 1H); 7.32 (br s, 1H); 7.56 (br s, 1H).

Example 32 (Second eluting diastereomer): 103.3 mg, 27% MS: 302 ES + (C ₁₂ H ₁₆ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 1.27 (t, 3H); 1.81 (m, 3H); 3.21 (m, 1H); 3.31 (m, 1H); 3.82 (m, 1H); 4.24 (m, 1H); 4.28 (q, 2H); 5.52 (m, 1H); 6.17 (m, 1H); 7.32 (br s, 1H); 7.55 (br s, 1H).

The stereochemistry of the two diastereomers was determined based on the order of their elution, as well as on the basis of the inhibitory activity of the corresponding carboxylic acids (examples 33 and 34): the more active acid corresponding to the first eluted diastereomer was designated as the R isomer.

Example 33. (2R) -2 - (((2S, 5R) -2-Carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy ) -2-fluoroacetic acid lithium salt

To a solution of (2R) -ethyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy ) -2-fluoroacetate (Example 31, 96.6 mg, 0.32 mmol) in THF (3 ml) and water (1 ml), lithium hydroxide (0.337 ml, 0.34 mmol) was added at 0 ° C. The reaction mixture was kept in an ice bath and stirred for 15 minutes. Added another 0.2 eq. lithium hydroxide. After 15 minutes, the pH of the reaction mixture was adjusted to pH = 7 with 0.5N hydrochloric acid. HCl. THF was evaporated and the remaining aqueous solution was frozen and lyophilized to give a pale yellow solid. After reverse phase HPLC and lyophilization (YMC Carotenoid C30, 19 mm x 150 mm, 5 μm, coupled to Synergi Polar RP 21.2 mm x 100 mm, 4 μm, 0-40% acetonitrile in water, 20 ml / min , 15 min) gave the title compound as a white solid, 34.8 mg, 40%. MS: 274 ES + (CioHi2FN30 ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 1.83 (m, 3H); 3.21 (m, 2H); 3.91 (m, 1H); 4.16 (m, 1H); 5.33 (m, 1H); 5.44 (m, 1H); 7.27 (br s, 1H); 7.53 (br s, 1H).

Example 34. (2S) -2 - (((2S, 5R) -2-Carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy ) -2-fluoroacetic acid lithium salt

The title compound was prepared from (2S) -ethyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-ene-6- yl) oxy) -2-fluoroacetate (Example 32, 91.8 mg, 0.30 mmol) according to the procedure for Example 33. Purification conditions were similar; after lyophilization, an off-white solid was obtained, 11.7 mg, 14%. MS: 274 ES + (C ₁₀ H ₁₂ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 1.81 (m, 3H); 3.21 (m, 2H); 3.87 (m, 1H); 4.16 (m, 1H); 5.25 (m, 1H); 5.45 (m, 1H); 7.28 (br s, 1H); 7.54 (br s, 1H).

Example 35. (2R) -Isopropyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl) oxy ) -2-fluoroacetate

Example 36. (2S) -Isopropyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl) oxy ) -2-fluoroacetate

To a solution of (2S, 5R) -6- (αlyloxy) -4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 45, 0.15 g , 0.63 mmol) in methanol (3 ml) at room temperature were added 1,3-dimethylbarbituric acid (0.197 g, 1.26 mmol) and tetrakis (triphenylphosphine) palladium (0) (0.073 g, 0, 06 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated to give an orange film. The orange film was dissolved in DMF (4 ml). Potassium carbonate (0.175 g, 1.26 mmol) and isopropyl 2-bromo-2-fluoroacetate (Intermediate 18, 0.377 g, 1.90 mmol) were added. The reaction mixture was stirred at room temperature for 4.5 h, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0-65% ethyl acetate / hexanes), a mixture of diastereomers in a ratio of 1: 1, 196 mg, 98% was obtained. Separation of diastereomers was carried out using reversed-phase HPLC (Atlantis T3, mm x 150 mm, 5 μm, 20-40% acetonitrile in water, 20 ml / min, 15 min). After lyophilization, both diastereomers were obtained as white solids.

Example 35 (First eluting diastereomer): 58.6 mg, 31%. MS: 316 ES + (Ci3Hi ₈ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.22 (m, 6H); 1.81 (m, 3H); 3.17 (m, 1H); 3.34 (m, 1H); 3.93 (m, 1H); 4.22 (m, 1H); 5.01 (q, 2H); 5.51 (m, 1H); 6.23 (m, 1H); 7.31 (br s, 1H); 7.54 (br s, 1H).

Example 36 (Second eluting diastereomer): 53.8 mg, 29%. MS: 316 ES + (C ₁₃ H ₁₈ FN3O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.28 (m, 6H); 1.81 (m, 3H); 3.19 (m, 1H); 3.29 (m, 1H); 3.82 (m, 1H); 4.24 (m, 1H); 5.06 (m, 1H); 5.52 (m, 1H); 6.14 (m, 1H); 7.32 (br s, 1H); 7.55 (br s, 1H).

Intermediates 46-50 were deliberately omitted.

Intermediate 51. (R) -tert-Butyl-4- (cyclopropyl (hydroxy) methyl) -2,2-dimethyloxazolidine-3-carboxylate boc

Cyclopropylmagnesium bromide was added to a solution of (R) -tert-butyl 4-formyl-2,2-dimethyloxαzolidine-3-carboxylate (Aldrich, 12.44 g, 54.26 mmol) in THF (150 ml) at -78 ° C (217 ml, 108.52 mmol) dropwise. The reaction mixture was allowed to warm to room temperature and was stirred overnight. The reaction was stopped by adding water and diluted with ethyl acetate and brine. The resulting emulsion was filtered through celite and the layers were separated. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-20% ethyl acetate / hexanes) afforded the title compound as a light yellow oil (12.47 g, 85%).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.16 (m, 2H); 0.37 (m, 2H); 0.82 (m, 1H); 1.45 (m, 15H); 2.87 (m, 1H); 3.86 (m, 2H); 3.97 (m, 1H); 4.74 (m, 1H).

Intermediate 52. (R) -tert-Butyl-4- (cyclopropanecarbonyl) -2,2-dimethyloxazolidine 3-carboxylate

Oud 'v boc

To a solution of (R) -tert-butyl 4- (cyclopropyl (hydroxy) methyl) -2,2-dimethyloxazolidine-3-carboxylate (Intermediate 51, 12.47 g, 45.95 mmol) in DCM (300 ml) Dess-Martin periodate (29.2 g, 68.93 mmol) was added at room temperature. The reaction mixture was stirred overnight, then diluted with ethyl acetate and washed with saturated sodium bicarbonate solution. An emulsion formed, which was filtered through celite. The layers were separated and the organics were washed with brine. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-20% ethyl acetate / hexanes) afforded the title compound as a colorless oil (11.15 g, 90%).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.90 (m, 4H); 1.38 (m, 12H); 1.54 (m, 3H); 2.12 (m, 1H); 3.94 (m, 1H); 4.18 (m, 1H); 4.56 (m, 1H).

Intermediate 53. (S) -tert-Butyl-4- (1-cyclopropylvinyl) -2,2-dimethyloxazolidine-3carboxylate

To a suspension of potassium tert-butylate (9.29 g, 82.80 mmol) in ether (250 mL) was added methyltriphenylphosphonium bromide (29.6 g, 82.80 mmol) at room temperature. The mixture turned a bright yellow color and was heated to 40 ° C for 1 hour.The mixture was cooled to room temperature and a solution of (R) -tert-butyl-4- (cyclopropαncarbonyl) -2,2-dimethyloxαzolidine-3-carboxylate was added (Intermediate 52, 11.15 g, 41.40 mmol) in ether (30 ml) and the reaction mixture was stirred for 2 h. The reaction was stopped by the addition of water (10 ml) and the layers were separated. The aqueous layer was extracted once with ether. The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-15% ethyl acetate / hexanes) afforded the title compound as a colorless

- 61 037916 oils (9.84 g, 89%). NMR (300 MHz, DMSO-d ₆ ) δ: 0.42 (m, 2H); 0.65 (m, 2H); 1.43 (m, 16H); 3.76 (m,

1H); 4.09 (m, 1H); 4.27 (m, 1H); 4.66 (m, 2H).

Intermediate 54. (S) -tert-Butyl-1- (tert-butyldimethylsilyloxy) -3-cyclopropylbut-3-en-2-ylcarbamate

HN, ^V boc

To a solution of (S) -tert-butyl 4- (1-cyclopropylvinyl) -2,2-dimethyloxazolidine-3-carboxylate (Intermediate 53, 8.25 g, 30.86 mmol) in methanol (100 ml) at room temperature, p-toluenesulfonic acid monohydrate (1.174 g, 6.17 mmol) was added. The reaction mixture was heated to 80 ° C overnight. Added another 0.2 eq. p-toluenesulfonic acid monohydrate was added and heated at 80 ° C for another 2 hours. The reaction mixture was cooled to room temperature. Triethylamine (4.29 ml, 30.86 mmol) and di-tert-butyl dicarbonate (3.37 g, 15.43 mmol) were added. The reaction mixture was stirred for two days and then concentrated. The residue was dissolved in ethyl acetate and washed once with saturated sodium bicarbonate solution. The combined organic extracts were dried over magnesium sulfate, filtered and concentrated. The resulting oil was dissolved in DCM (100 ml). Imidazole (2.73 g, 40.11 mmol), 4-dimethylaminopyridine (0.754 g, 6.17 mmol) and tert-butyldimethylsilyl chloride (4.65 g, 30.86 mmol) were added and the reaction mixture was stirred overnight at room temperature. The reaction mixture was filtered to remove solids and washed twice with brine. The organic layer was dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-10% ethyl acetate / hexanes) afforded the title compound as a colorless oil (6.77 g, 64%). MS: 342 ES + (C ₁₈ H ₃₅ NO ₃ Si) 1H NMR (300 MHz, DMSO-d6) δ: 0.04 (s, 6H); 0.39 (m, 2H); 0.63 (m, 2H); 0.85 (s, 9H); 1.32 (m, 1H); 1.37 (m, 9H); 3.55 (m, 1H); 3.67 (m, 1H); 3.99 (m, 1H); 4.63 (s, 1H); 4.78 (s, 1H); 6.80 (m, 1H).

Intermediate 55. (S) -1- (tert-Butylgimethylsilyloxy) -3-cyclopropylbut-3-en-2amine

TVZO ^ A ^ Ts-7 nh ₂ ^v

To a solution of (S) -tert-butyl-1 - (tert-butyldimethylsilyloxy) -3-cyclopropylbut-3-en-2-yl carbamate (Intermediate 54, 6.77 g, 19.82 mmol) in DCM (100 ml) zinc bromide (17.86 g, 79.28 mmol) was added at room temperature. The reaction mixture was stirred overnight at room temperature. Added another 1 eq. zinc bromide. After a few hours, the reaction mixture was filtered and washed with saturated sodium bicarbonate solution. The resulting emulsion was filtered through a nylon filter and the layers were separated. The organics were dried over magnesium sulfate, filtered and concentrated to give the title compound as a yellow oil (4.61 g, 96%).

1H NMR (300 MHz, DMSO-d6) δ: 0.04 (s, 6H); 0.39 (m, 2H); 0.63 (m, 2H); 0.87 (s, 9H); 1.35 (m, 1H); 1.81 (m, 2H); 3.33 (m, 1H); 3.45 (m, 1H); 3.67 (m, 1H); 4.59 (s, 1H); 4.83 (m, 1H).

Intermediate 56. (S) -tert-Butyl-1- (tert-butylgimethylsilyloxy) -3-cyclopropylbut-3-en-2-yl (2- (methoxy (methyl) amino) -2-oxoethyl) carbamate meso '^ V ^ y ^

Boc ^A >

I

The title compound was prepared from (S) -1- (tert-butylgimethylsilyloxy) -3-cyclopropylbut-3-en-2-amine (Intermediate 55, 4.61 g, 19.09 mmol) and 2-bromo-N α-methoxy-Nmethylacetamide (Intermediate 4, 3.16 g, 17.36 mmol) as described for Intermediate 5. The desired product was obtained as a light yellow oil (4.94 g, 64%). MS: 443 ES + (C22H42N2OsSi).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.03 (m, 6H); 0.35 (m, 1H); 0.48 (m, 1H); 0.61 (m, 2H); 0.83 (m, 9H); 1.35 (m, 9H); 3.07 (m, 3H); 3.65 (m, 3H); 3.84 (m, 2H); 4.02 (m, 2H); 4.54 (m, 1H); 4.83 (m, 2H).

Intermediate 57. (S) -tert-Butyl-1- (tert-butylgimethylsilyloxy) -3-cyclopropylbut-3-en-2-yl (2-oxopent-3-yl) carbamate

A suspension of cerium (III) chloride (27.8 g, 112.95 mmol) in THF (100 ml) was vigorously stirred at room temperature for 2 hours.The suspension was cooled to -78 ° C and (E) - 62 037916 was added dropwise prop-1-enylmagnesium bromide (0.5 M in THF) (226 ml, 112.95 mmol). The mixture was stirred at -78 ° C for 1.5 h. Then (S) -tert-butyl-1- (tert-butyldimethylsilyloxy) -3-cyclopropylbut-3-en-2-yl (2- (methoxy (methyl) amino) -2-oxoethyl) carbamate (Intermediate 56, 5 g, 11.30 mmol) in THF (20 ml) at -78 ° C. The reaction mixture was stirred at -78 ° C for 30 min and then warmed to 0 ° C over 15 min. The reaction was stopped by the addition of 10% citric acid, further diluted with water and extracted twice with ether. The organics were washed once with brine, dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0% -20% ethyl acetate / hexanes) afforded the title compound as a light yellow oil (4.0 g, 84%). MS: 424 ES + (C ₂₃ H ₄₁ NO ₄ Si).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.03 (m, 6H); 0.43 (m, 2H); 0.61 (m, 2H); 0.83 (m, 9H); 1.34 (m, 10H); 1.84 (m, 2H); 2.04 (m, 1H); 3.74 (m, 1H); 3.84 (m, 2H); 4.03 (m, 1H); 4.57 (m, 1H); 4.79 (m, 2H); 6.28 (m, 1H); 6.84 (m, 1H).

Intermediate 58. (S) -tert-Butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -3-cyclopropyl-5-oxo-5,6-dihydropyridine-1 (2H) -carboxylate

The title compound was prepared from (S) -tert-butyl-1- (tert-butyldimethylsilyloxy) -3cyclopropylbut-3-en-2-yl (2-oxopent-3-enyl) carbamate (Intermediate 57, 4g, 9 , 44 mmol) according to the procedure described for Intermediate 7, with the difference that the reaction mixture was heated at 110 ° C overnight. The desired product was obtained as a light brown oil (2.97 g, 82%). MS: 382 ES + (C ₂₀ H ₃₅ NO ₄ Si).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.01 (m, 6H); 0.62 (m, 1H); 0.80 (s, 9H); 1.00 (m, 3H); 1.42 (s, 9H); 1.61 (m, 1H); 3.80 (m, 1H); 3.95 (m, 2H); 4.19 (m, 1H); 4.75 (m, 1H); 5.72 (s, 1H).

Intermediate 59. (2S, 5S) -tert-butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -3cyclopropyl-5-hydroxy-5,6-dihydropyridine-1 (2H) -carboxylate

BUT

TBSO boc ^{/ Nx} ^ 'ΌΗ

The title compound was prepared from (S) -tert-butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -3-cyclopropyl-5-oxo-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 58, 2.97 g, 7.78 mmol) as described for Intermediate 10. The desired product was obtained as a brownish beige oil (2.74 g, 92%). MS: 384 ES + (C ₂₀ H ₃₇ NO ₄ Si).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.02 (m, 6H); 0.34 (m, 1H); 0.47 (m, 1H); 0.64 (m, 2H); 0.85 (m, 9H); 1.26 (m, 1H); 1.39 (s, 9H); 2.65 (m, 1H); 3.89 (m, 3H); 4.05 (m, 1H); 4.95 (m, 1H); 5.34 (m, 1H).

Intermediate 60. (2S, 5R) -tert-butyl-5- (N- (αlyloxy) -2-nitrophenylsulfonamido) 2 - ((tert-butyldimethylsilyloxy) methyl) -3-cyclopropyl-5,6-dihydropyridine-1 ( 2H) -carboxylate

L tbso '' (η

I

Ns

The title compound was prepared from (2S, 5S) -tert-butyl-2 - ((tert-butyldimethylsilyloxy) methyl) -3-cyclopropyl-5-hydroxy-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 59, 2.74 g, 7.14 mmol) and N- (αlyloxy) -2-nitrobenzenesulfonamide (1.85 g, 7.14 mmol) according to the procedure described for Intermediate 11. The target product was obtained as a light yellow oil ( 3.19 g, 71%). MS: 624 ES + (C ₂ cH ₄ 5N ₃ O ₈ SSi).

1H NMR (300 MHz, DMSO-d6) δ: 0.00 (m, 6H); 0.34 (m, 1H); 0.63 (m, 2H); 0.83 (m, 9H); 1.37 (m, 9H); 3.30 (m, 1H); 3.84 (m, 2H); 4.30 (m, 4H); 5.18 (m, 2H); 5.75 (m, 1H); 8.04 (m, 4H).

Intermediate 61. (2S, 5R) -tert-Butyl 5- (N- (αlyloxy) -2-nitrophenylsulfonamido) 3-cyclopropyl-2- (hydroxymethyl) -5,6-dihydropyridine-1 (2H) -carboxylate boc ^x 'X'n

Ns

The title compound was prepared from (2S, 5R) -tert-butyl-5- (N- (allyloxy) -2-nitrophenylsulfonamido) -2 - ((tert-butyldimethylsilyloxy) methyl) -3-cyclopropyl-5,6-dihydropyridine -1 (2H) carboxylate (Intermediate 60, 3.19 g, 5.11 mmol) as described for Intermediate 12. The desired product was obtained as a brownish-beige foam (2.35 g, 90%).

- 63 037916

MS: 510 ES + (C2 ₃ H ₃₁ N ₃ O8S).

1H NMR (300 MHz, DMSO-d6) δ: 0.32 (m, 2H); 0.62 (m, 2H); 1.35 (m, 9H); 3.30 (m, 1H); 3.67 (m, 2H);

4.27 (m, 4H); 4.71 (m, 1H); 5.19 (m, 2H); 5.71 (m, 1H); 8.04 (m, 4H).

Intermediate 62. (2S, 5R) -5- (N- (Allyloxy) -2-nitrophenylsulfonamido) -1- (tert-butoxycarbonyl) -3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxylic acid

-Oh

Ns

The title compound was prepared from (2S, 5R) -tert-butyl-5- (N- (allyloxy) -2-nitrophenylsulfonamido) -3-cyclopropyl-2- (hydroxymethyl) -5,6-dihydropyridine-1 (2H) β-carboxylate (Intermediate 61, 2.35 g, 4.61 mmol) as described for Intermediate 13. The desired product was obtained as an orange foam (2.28 g, 94%). MS: 524 ES + (C ₂₃ H ₂₉ N ₃ O ₉ S).

Intermediate 63. (2S, 5R) -tert-Butyl 5- (N- (allyloxy) -2-nitrophenylsulfonamido) 2-carbamoyl-3-cyclopropyl-5,6-dihydropyridine-1 (2H) -carboxylate

..Oh

Boc ^^ M ' ⁰ Ns

The title compound was prepared from (2S, 5R) -5- (N- (allyloxy) -2-nitrophenylsulfonamido) -1- (tert-butoxycarbonyl) -3-cyclopropyl-1,2,5,6-tetrahydropyridine-2- carboxylic acid (Intermediate 62, 2.28 g, 4.35 mmol) according to the procedure described for Intermediate 14. The desired product was obtained as an orange foam (1.07 g, 47%). MS: 523 ES + (C23H30N4O8S).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 0.23 (m, 2H); 0.59 (m, 2H); 1.35 (m, 9H); 3.58 (m, 1H); 4.23 (m, 3H); 4.72 (m, 1H); 5.19 (m, 2H); 5.71 (m, 1H); 7.18 (m, 1H); 7.59 (m, 1H); 8.04 (m, 4H).

Intermediate 64. (2S, 5R) -5- (N- (Allyloxy) -2-nitrophenylsulfonamido) -3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide d, aX

Ns

The title compound was prepared from (2S, 5R) -tert-butyl 5- (N- (allyloxy) -2nitrophenylsulfonamido) -2-carbamoyl-3-cyclopropyl-5,6-dihydropyridine-1 (2H) -carboxylate (Intermediate compound 63, 0.932 g, 1.78 mmol) as described for intermediate 25. The desired product was obtained as an orange foam (0.518 g, 68%). MS: 423 ES + (C ₁₈ H ₂₂ N ₄ O ₆ S).

¹ H NMR (300 MHz, DMSO-d6) δ: 0.18 (m, 2H); 0.53 (m, 2H); 1.29 (m, 1H); 2.30 (m, 1H); 2.58 (m, 1H); 2.95 (m, 1H); 3.72 (m, 1H); 4.22 (m, 1H); 4.36 (m, 2H); 4.96 (m, 1H); 5.24 (m, 2H); 5.80 (m, 1H); 7.07 (bs, 1H); 7.39 (bs, 1H); 8.04 (m, 4H).

Intermediate 65. (R) -5- (Allyloxyamino) -3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide

The title compound was prepared from (2S, 5R) -5- (N- (allyloxy) -2-nitrophenylsulfonamido) -3-cyclopropyl-1,2,5,6-tetrahydropyridine-2-carboxamide (Intermediate 64, 0.518 g , 1.23 mmol) as described for Intermediate 26. The desired product was obtained as a light yellow oil (0.171 g, 59%). The product was a mixture of diastereomers. MS: 238 ES + (C ^ H ^ N ^).

1H NMR (300 MHz, DMSO-d6) δ: 0.28 (m, 2H); 0.41 (m, 2H); 0.54 (m, 2H); 1.33 (m, 1H); 2.49 (m, 1H); 2.64 (m, 1H); 2.93 (m, 1H); 3.23 (m, 1H); 3.65 (m, 1H); 4.07 (m, 2H); 5.19 (m, 3H); 5.89 (m, 1H); 6.26 (m, 1H); 6.97 (bs, 1H); 7.34 (bs, 1H).

Intermediate 66. (2S, 5R) -6- (Allyloxy) -3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

- 64 037916

The title compound was prepared from (R) -5- (allyloxyamino) -3-cyclopropyl-1,2,5,6tetrahydropyridine-2-carboxamide (Intermediate 65, 0.316 g, 1.33 mmol) using the procedure outlined for Intermediate compounds 27. The target product was obtained as a colorless oil (0.261 g, 74%). MS: 264 ES + (C13H17N3O3).

¹ H NMR (300 MHz, DMSO-ds) δ: 0.37 (m, 2H); 0.60 (m, 2H); 1.20 (m, 1H); 2.98 (m, 1H); 3.79 (m, 1H); 3.92 (m, 1H); 4.20 (m, 1H); 4.33 (m, 2H); 5.28 (m, 2H); 5.93 (m, 2H); 7.30 (bs, 1H); 7.86 (bs, 1H).

Example 37. (2R) -Isopropyl-2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl ) oxy) -2-fluoro acetate

Example 38. (2S) -Isopropyl-2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl ) oxy) -2-fluoro acetate

To a solution of (2S, 5R) -6- (αlyloxy) -3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 66, 0.15 g , 0.57 mmol) in methanol (3 ml), 1,3-dimethylbarbituric acid (0.178 g, 1.14 mmol) and tetrakis (triphenylphosphine) palladium (0) (0.066 g, 0.06 mmol) were added at room temperature. The reaction mixture was stirred at room temperature for 2 hours, then concentrated to give an orange film. The orange film was dissolved in DMF (4 ml). Potassium carbonate (0.157 g, 1.14 mmol) and isopropyl 2-bromo-2-fluoroacetate (Intermediate 18, 0.340 g, 1.71 mmol) were added. The reaction mixture was stirred at room temperature overnight, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-65% ethyl acetate / hexanes) gave a 1: 1 mixture of diastereomers, 166.3 mg, 86%. Separation of diastereomers was performed using reverse phase HPLC (Atlantis T3, 19 mm x 150 mm, 5 μm, 20-40% acetonitrile in water, 20 ml / min, 15 min). After lyophilization, both diastereomers were obtained as white solids.

Example 37 (First eluting diastereomer): 47.3 mg, 24%. MS: 342 ES + (C ₁₅ H ₂₀ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 0.39 (m, 2H); 0.61 (m, 1H); 1.19 (d, 3H); 1.21 (m, 1H); 1.24 (d, 3H); 2.99 (m, 1H); 3.88 (m, 1H); 4.01 (m, 1H); 4.29 (m, 1H); 5.00 (m, 1H); 5.87 (m, 1H); 6.20 (m, 1H); 7.36 (br s, 1H); 7.90 (br s, 1H).

Example 38 (Second eluting diastereomer): 49.8 mg, 26%. MS: 342 ES + (C15H20FN3O5).

1H NMR (300 MHz, DMSO-d6) δ: 0.41 (m, 2H); 0.62 (m, 1H); 1.22 (m, 1H); 1.27 (d, 3H); 1.29 (d, 3H); 3.03 (m, 1H); 3.91 (m, 1H); 3.94 (m, 1H); 4.31 (m, 1H); 5.05 (m, 1H); 5.91 (m, 1H); 6.12 (m, 1H); 7.37 (br s, 1H); 7.93 (br s, 1H).

Example 39. (2R) -Ethyl-2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-en-6-yl) oxy ) -2-fluoroacetate

Example 40. (2S) -Ethyl-2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl) oxy ) -2-fluoroacetate

To a solution of (2S, 5R) -6- (αlyloxy) -3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-carbo- 65 037916 xamide (Intermediate 66, 0.2972 g, 1.13 mmol) in methanol (6 ml) at room temperature were added 1,3-dimethylbarbituric acid (0.352 g, 2.26 mmol) and tetrakis (triphenylphosphine) palladium (0) (0.130 g, 0, 11 mmol). The reaction mixture was stirred at room temperature for 2 hours, then concentrated to give an orange film. The orange film was dissolved in DMF (6 ml). Potassium carbonate (0.468 g, 3.39 mmol) and ethyl bromofluoroacetate (0.534 ml, 4.52 mmol) were added. The reaction mixture was stirred overnight at room temperature, then diluted with ethyl acetate and filtered through a 0.45 μm filter to remove solid potassium carbonate. The filtrate was washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (065% ethyl acetate / hexanes), a 1: 1 mixture of diastereomers, 303.7 mg, 82% was obtained. Separation of diastereomers was performed using reverse phase HPLC (Atlantis T3, 19 mm x 150 mm, 5 μm, 20% -40% acetonitrile in water, 20 ml / min, 15 min). After lyophilization, both diastereomers were obtained as white solids.

Example 39 (First eluted diastereomer): 107 mg, 29% MS: 328 ES + (C ₁₄ H ₁₈ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-db) δ: 0.39 (m, 2H); 0.61 (m, 2H); 1.21 (m, 4H); 3.01 (m, 1H); 3.89 (m, 1H); 4.02 (m, 1H); 4.19 (m, 2H); 4.29 (s, 1H); 5.88 (m, 1H); 6.22 (m, 1H); 7.36 (br s, 1H); 7.91 (br s, 1H).

Example 40 (Second eluting diastereomer): 110.9 mg, 30%. MS: 328 ES + (C ₁₄ H ₁₈ FN ₃ O ₅ ). NMR (300 MHz, DMSO-d ₆ ) δ: 0.40 (m, 2H); 0.61 (m, 2H); 1.26 (m, 4H); 3.04 (m, 1H); 3.90 (m, 1H);

3.94 (m, 1H); 4.26 (m, 3H); 5.90 (m, 1H); 6.24 (m, 1H); 7.36 (br s, 1H); 7.92 (br s, 1H).

Example 41. (2R) -2 - (((2S, 5R) -2-Carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy) - 2-fluoroacetic acid lithium salt

To a solution of (2R) -ethyl-2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl) oxy) -2-fluoroacetate (Example 39, 96.6 mg, 0.30 mmol) in THF (3 ml) and water (1 ml) at 0 ° C was added lithium hydroxide (1M) (0.310 ml, 0.31 mmol). The reaction mixture was kept in an ice bath and stirred for 15 minutes. Added another 0.2 eq. lithium hydroxide. After 15 minutes, the pH of the reaction mixture was adjusted to pH = 7 with 0.5N hydrochloric acid. HCl. The mixture was frozen and lyophilized to give a pale yellow solid, 90.4 mg. After reverse phase HPLC (YMC Carotenoid 30, 19 mm x 150 mm, 5 μm, coupled to Synergi Polar RP, 21.2 mm x 100 mm, 4 μm, 0% 25% acetonitrile in water, 20 ml / min, 5 min ) gave the title compound as a white solid, 45.9 mg, 52%. MS: 300 ES + (C ₁₂ H ₁₄ FN ₃ O ₅ ).

NMR (300 MHz, DMSO-d ₆ ) δ: 0.38 (m, 2H); 0.59 (m, 2H); 1.20 (m, 1H); 3.02 (m, 1H); 3.83 (m, 1H); 4.00 (m, 1H); 4.25 (m, 1H); 5.24 (m, 1H); 5.89 (m, 1H); 7.30 (br s, 1H); 7.88 (br s, 1H).

Example 42. (2R) -2 - (((2S, 5R) -2-Carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy) - 2-fluoroacetic acid lithium salt

To a solution of (2S) -2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy) -2 β-fluoroacetic acid (Example 40, 96.6 mg, 0.30 mmol) in THF (3 ml) and water (1) at 0 ° C was added lithium hydroxide (1M) (0.310 ml, 0.31 mmol). The reaction mixture was kept in an ice bath and stirred for 15 minutes. Added another 0.2 eq. lithium hydroxide. After 15 minutes, the pH of the reaction mixture was adjusted to pH 7 with 0.5N hydrochloric acid. HCl. The mixture was frozen and lyophilized to give a pale yellow solid, 90.6 mg. After reverse phase HPLC (YMC Carotenoid 30, 19 mmx150 mm, 5 μm, coupled with Synergi Polar RP, 21.2 mmx100 mm, 4 μm, 025% acetonitrile in water, 20 ml / min, 5 min) the title compound as a white solid, 41.2 mg, 45%. MS: 300 ES + (C12H1 ₄ FN ₃ O ₅ ) Ή NMR (300 MHz, DMSO-de) δ: 0.39 (m, 2H); 0.59 (m, 2H); 1.20 (m, 1H); 3.02 (m, 1H); 3.82 (m, 1H); 3.98 (m, 1H); 4.24 (m, 1H); 5.24 (m, 1H); 5.90 (m, 1H); 7.31 (br s, 1H); 7.90 (br s, 1H).

Example 43.2 - (((2S, 5R) -2-Carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2- fluoroacetic acid lithium salt

- 66 037916

(2R) -2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetic acid (example 41, 8 mg, 0.03 mmol) and (2S) -2 - (((2S, 5R) -2-carbamoyl-3-cyclopropyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl) oxy) -2-fluoroacetic acid (Example 42, 8 mg, 0.03 mmol) was combined in an amber tube. Water (1.5 ml) was added. The mixture was frozen and lyophilized to give a white solid, 16 mg. MS: 300 ES + (C ₁₂ H ₁₄ FN ₃ O ₅ ).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 0.39 (m, 2H); 0.58 (m, 2H); 1.19 (m, 1H); 3.00 (m, 1H); 3.81 (m, 1H); 3.98 (m, 1H); 4.24 (m, 1H); 5.19 (m, 1H); 5.89 (m, 1H); 7.29 (br s, 1H); 7.88 (br s, 1H).

Intermediate 67. (1-Isopropyl-2-methylpropyl) -2-bromo-2,2-difluoroacetate

To a solution of 2,4-dimethylpentan-3-ol (0.72 ml, 5.17 mmol) and N, N-diisopropylethylamine (1.81 ml, 10.34 mmol) in DCM (20 ml) at 0 ° С 2-bromo-2,2-difluoroacetyl chloride (0.49 mL, 5.17 mmol) was added dropwise. The reaction mixture was stirred at 35 ° C overnight. The reaction was stopped by adding 10 ml of 1N hydrochloric acid. of hydrochloric acid. The layers were separated. The organics were washed twice with water, once with brine, then dried over magnesium sulfate, filtered and concentrated to give the title compound as a dark orange oil, 1.95 g, quant. The crude material was used in the next step.

¹ H NMR (300 MHz, CDCl ₃ -d) δ: 0.95 (m, 12H); 2.06 (m, 2H); 4.74 (m, 1H).

Example 44. (1-Isopropyl-2-methylpropyl) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -6-yl] oxy] -2,2-difluoroacetate

To a solution of (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 150 mg, 0.76 mmol) in DMF (5 ml) at room temperature were added potassium carbonate (210.27 mg, 1.52 mmol) and (1-isopropyl-2-methylpropyl) 2-bromo-2,2-difluoroacetate (Intermediate 67, 623 , 28 mg, 2.28 mmol). The reaction mixture was stirred for 4 hours at room temperature. Two more equivalents of (1-isopropyl-2-methylpropyl) 2-bromo-2,2-difluoroacetate were added and the reaction was stirred overnight at room temperature. The reaction mixture was diluted with ethyl acetate and filtered to remove potassium carbonate. The filtrate was washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0% -15% acetone / dichloromethane) and lyophilization, the title compound was obtained as a light orange sticky film, 44.6 mg, 13%. MS: 390 eS + (C ₁₇ H ₂ 5FN ₃ O ₅ ).

¹ H NMR (300 MHz, DMSO-d6) δ: 0.86 (m, 12H); 1.64 (s, 3H); 1.99 (m, 2H); 3.15 (m, 1H); 3.85 (m, 1H); 4.07 (m, 1H); 4.28 (s, 1H); 4.69 (m, 1H); 6.03 (m, 1H); 7.42 (s, 1H); 7.85 (s, 1H).

Intermediate 68. Octyl- (2R) -2-bromo-2-fluoroacetate

To a suspension of (2R) -2-bromo-2-fluoroacetic acid; (1S) -1-phenylethanamine (Intermediate 168, 642.8 mg, 2.31 mmol) and 1-octanol (554 mg, 5.78 mmol) in DCM (9 ml) was added dropwise chlorotrimethylsilane (1 , 19 ml, 9.39 mmol). The suspension turned into a solution and was stirred overnight at room temperature. The reaction mixture was washed three times with water. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-10% ethyl acetate / hexanes) afforded the title compound as a colorless liquid, 554.9 mg, 89%.

1H NMR (300 MHz, CDCl ₃ -d) δ: 0.84 (m, 3H); 1.24 (m, 10H); 1.61 (m, 2H); 4.22 (m, 2H); 7.25 (d, 1H).

Example 45. Octyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy ] -2-fluoroacetate

- 67 037916

To a solution of (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 150 mg, 0.76 mmol) in 1,4-dioxane (4 ml) and DMF (0.5 ml) was added octyl- (2R) -2-bromo-2-fluoroacetate (Intermediate 68, 0.09 ml, 2.06 mmol). The reaction mixture was cooled to 0 ° C and DBU (0.11 mL, 0.76 mmol) was added dropwise. The reaction mixture was stirred for 10 min, then diluted with ethyl acetate and washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated to give an orange oil. Chromatography on silica gel (0-60% ethyl acetate / hexanes) afforded the title compound as a white solid, 247.9 mg, 84%. MS: 386 ES + (C18H28FN3O5).

1H NMR (300 MHz, DMSO-ds) δ: 0.86 (m, 3H); 1.25 (m, 10H); 1.59 (m, 5H); 3.04 (m, 1H); 3.78 (m, 1H); 4.04 (m, 1H); 4.16 (m, 3H); 6.00 (m, 1H); 6.24 (d, 1H); 7.37 (s, 1H); 7.81 (s, 1H).

Intermediate 69. Octyl- (2R) -2-bromo-2-fluoroacetate

Ά

O

The title compound was prepared from (2R) -2-bromo-2-fluoroacetic acid; (1S) -1phenylethanamine (Intermediate 168, 526.2 mg, 1.89 mmol) and methanol (260 mg, 5.68 mmol) according to the procedure for Intermediate 68 to give a white oil / solid, 260 mg, 80% ...

1 H NMR (300 MHz, CDCl ₃ -d) δ: 3.84 (s, 3H); 6.52 (d, 1H).

Example 46. Methyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy ] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 120 mg, 0 , 61 mmol) and methyl- (2R) -2-bromo-2-fluoroacetate (Intermediate 69, 0.09 ml, 1.52 mmol) according to the procedure for example 45 as a white foam, 81.2 mg, 46% ... MS: 288 ES + (C _n H ₁₄ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-ds) δ: 1.63 (s, 3H); 3.07 (m, 1H); 3.75 (m, 1H); 3.78 (s, 3H); 4.05 (m, 1H); 4.19 (s, 1H); 6.02 (m, 1H); 6.24 (m, 1H); 7.37 (s, 1H); 7.81 (s, 1H).

Intermediate 70. Allyl- (2R) -2-bromo-2-fluoroacetate

The title compound was prepared from (2R) -2-bromo-2-fluoroacetic acid; (1S) -1phenylethanamine (Intermediate 168, 510 mg, 1.83 mmol) and allyl alcohol (0.37 mL, 5.5 mmol) according to the procedure for Intermediate 68 to give a colorless liquid, 100 mg, 28%. 1H NMR (300 MHz. CDCl ₃ -d) δ: 4.71 (m, 2H); 5.31 (m, 2H); 5.86 (m, 1H); 6.55 (d, 1H).

Example 47. Allyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy ] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 50 mg, 0 , 25 mmol) and allyl- (2R) -2-bromo-2-fluoroacetate (Intermediate 70, 100 mg, 0.51 mmol) according to the procedure for example 45 to obtain a white foam, 61.3 mg, 77%. MS: 314 ES + (C ₁₃ H ₁₆ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.63 (s, 3H); 3.55 (m, 1H); 3.76 (m, 1H); 4.05 (m, 1H); 4.19 (s, 1H); 4.70 (m, 2H); 5.36 (m, 2H); 5.90 (m, 1H); 6.01 (m, 1H); 6.28 (d, 1H); 7.37 (s, 1H); 7.82 (s, 1H).

Intermediate 71. Propyl- (2R) -2-bromo-2-fluoroacetate

- 68 037916

The title compound was prepared from (2R) -2-bromo-2-fluoroacetic acid; (1S) -1phenylethanamine (Intermediate 168, 444.9 mg, 1.6 mmol) and 1-propanol (0.3 mL, 4 mmol) according to the procedure for Intermediate 68 to give a colorless liquid, 318 mg, 100%. Ή NMR (300 MHz, CDCl3-d) δ: 0.92 (t, 3H); 1.67 (m, 2H); 4.19 (m, 2H); 6.51 (d, 1H).

Example 48. Propyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy ] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 150 mg , 0.76 mmol) and propyl (2R) -2-bromo-2-fluoroacetate (Intermediate 71, 302.78 mg, 1.52 mmol) according to the procedure for example 45 to obtain a white sticky foam, 156 mg, 65% ... MS: 316 ES + (C ₁₃ H ₁₈ FN ₃ O ₅ ).

¹ H NMR (300 MHz, DMSO-d6) δ: 0.89 (t, 3H); 1.61 (m, 5H); 3.05 (m, 1H); 3.76 (m, 1H); 4.04 (m, 1H); 4.13 (m, 2H); 4.19 (m, 1H); 6.01 (m, 1H); 6.25 (d, 1H); 7.37 (s, 1H); 7.82 (s, 1H).

Intermediate 72. Isobutyl- (2R) -2-bromo-2-fluoroacetate

The title compound was prepared from (2R) -2-bromo-2-fluoroacetic acid; (1S) -1phenylethanamine (Intermediate 168, 487.5 mg, 1.75 mmol) and 2-methyl-1-propanol (0.4 mL, 4.38 mmol) according to the procedure for Intermediate 68 to give a colorless liquid, 373 mg, 100%. ¹ H NMR (300 MHz, CDCl ₃ -d) δ: 0.91 (d, 6H); 1.97 (m, 1H); 4.00 (m, 2H); 6.51 (d, 1H).

Example 49. Isobutyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl] oxy ] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 150 mg , 0.76 mmol) and isobutyl (2R) -2-bromo-2-fluoroacetate (Intermediate 72, 372.73 mg, 1.75 mmol) according to the procedure for example 45 to give a white sticky foam, 161 mg, 64% ... MS: 330 ES + (C ₁₄ H ₂₀ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 0.90 (d, 6H); 1.63 (s, 3H); 1.90 (m, 1H); 3.05 (m, 1H); 3.76 (m, 1H); 3.97 (m, 2H); 4.04 (m, 1H); 4.20 (s, 1H); 6.00 (m, 1H); 6.27 (d, 1H); 7.37 (s, 1H); 7.82 (s, 1H).

Intermediate 73. Butyl- (2R) -2-bromo-2-fluoroacetate

The title compound was prepared from (2R) -2-bromo-2-fluoroacetic acid; (1S) -1phenylethanamine (Intermediate 168, 400 mg, 1.44 mmol) and 1-butanol (0.33 mL, 3.6 mmol) according to the procedure for Intermediate 68 to give a colorless liquid, 322 mg, quant.

L NMR (300 MHz, CDCl ₃ -d) δ: 0.88 (t, 3H); 1.33 (m, 2H); 1.65 (m, 2H); 4.23 (m, 2H); 6.50 (d, 1H).

Example 50. Butyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy ] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 150 mg, 0 , 76 mmol) and butyl- (2R) -2-bromo-2-fluoroacetate (Intermediate 73, 322.49 mg, 1.51 mmol) according to the procedure for example 45 to obtain a white sticky foam, 198 mg, 79%. MS: 330 ES + (C ₁₄ H ₂₀ FN ₃ O ₅ ).

- 69 037916

1H NMR (300 MHz, DMSO-d6) δ: 0.88 (t, 3H); 1.35 (m, 2H); 1.59 (m, 2H); 1.63 (s, 3H); 3.06 (m, 1H);

3.77 (m, 1H); 4.04 (m, 1H); 4.20 (m, 2H); 4.21 (s, 1H); 6.00 (m, 1H); 6.24 (d, 1H); 7.37 (s, 1H); 7.82 (s, 1H).

Intermediate 74. Pentyl- (2R) -2-bromo-2-fluoroacetate

The title compound was prepared from (2R) -2-bromo-2-fluoroacetic acid; (1S) -1phenylethanamine (Intermediate 168, 474.2 mg, 1.71 mmol) and 1-pentanol (0.46 mL, 4.26 mmol) according to the procedure for Intermediate 68 to give a colorless liquid, 363 mg, 93 %.

1H NMR (300 MHz, CDCl3-d) δ: 0.88 (m, 3H); 1.28 (m, 4H); 1.60 (m, 2H); 4.17 (m, 2H); 6.45 (d, 1H).

Example 51. Pentyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy ] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 150 mg , 0.76 mmol) and pentyl (2R) -2-bromo-2-fluoroacetate (Intermediate 74, 362.73 mg, 1.6 mmol) according to the procedure for example 45 to give a sticky white foam, 225 mg, 86% ... MS: 344 ES + (C ₁₅ H ₂₂ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 0.87 (m, 3H); 1.30 (m, 4H); 1.58 (m, 2H); 1.63 (s, 3H); 3.04 (m, 1H); 3.77 (m, 1H); 4.04 (m, 1H); 4.20 (m, 2H); 4.21 (s, 1H); 6.00 (m, 1H); 6.24 (d, 1H); 7.37 (s, 1H); 7.82 (s, 1H).

Intermediate 75. Hexyl- (2R) -2-bromo-2-fluoroacetate

The title compound was prepared from (2R) -2-bromo-2-fluoroacetic acid; (1S) -1phenylethanamine (Intermediate 168, 400 mg, 1.44 mmol) and hexyl alcohol (0.45 mL, 3.6 mmol) according to the procedure for Intermediate 68 to give a colorless liquid, 313 mg, 90%.

1H NMR (300 MHz. CDCl ₃ -d) δ: 0.86 (m, 3H); 1.22 (m, 6H); 1.59 (m, 2H); 4.17 (m, 2H); 6.45 (d, 1H).

Example 52 Hexyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en6-yl] oxy ] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 193, 150 mg , 0.76 mmol) and hexyl (2R) -2-bromo-2-fluoroacetate (Intermediate 75, 313.62 mg, 1.3 mmol) according to the procedure for Example 45 to obtain a sticky white foam, 224 mg, 82% ... MS: 358 ES + (C ₁₆ H ₂₄ FN ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 0.87 (m, 3H); 1.30 (m, 6H); 1.57 (m, 2H); 1.62 (s, 3H); 3.04 (m, 1H); 3.77 (m, 1H); 4.04 (m, 1H); 4.19 (m, 2H); 4.20 (s, 1H); 6.00 (m, 1H); 6.24 (d, 1H); 7.37 (s, 1H); 7.82 (s, 1H).

Intermediate 76.1-Chloroethylisopropyl carbonate

ITI ¹ about ¹

To a solution of 2-propanol (0.64 ml, 8.39 mmol) and pyridine (0.79 ml, 9.79 mmol) at -78 ° C was added 1-chloroethyl chloroformate (0.76 ml, 6.99 mmol ). The reaction mixture was allowed to slowly warm to room temperature and stirred overnight. The reaction mixture turned into a solid white clot and was sonicated in dichloromethane. The resulting slurry was concentrated and the white solid was dissolved in ethyl acetate and washed with water and brine. The organics were dried over magnesium sulfate, filtered and concentrated to give the title compound as a colorless liquid, 1.29 g, 99%. 1H NMR (300 MHz, CDCl ₃ -d) δ: 1.35 (m, 6H); 1.84 (d, 3H); 4.96 (m, 1H); 4.44 (m, 1H).

Example 53 1-Isopropoxycarbonyloxyethyl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6-yl ] oxy] -2-fluoroacetate

- 70 037916

To a solution of (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetic acid (example 4, 194.46 mg, 0.71 mmol), N, N-diisopropylethylamine (0.12 ml, 0.71 mmol) and 1-chloroethyl isopropyl carbonate (Intermediate 76, 237.15 mg, 1.42 mmol) in DMF (5 ml) was added tetrabutyl ammonium chloride (162.22 mg, 0.71 mmol) at room temperature. The reaction mixture was heated at 40 ° C for about 4 hours, then diluted with ethyl acetate and washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated to give an orange oil. Chromatography on silica gel (0% -50% ethyl acetate / hexanes) afforded the title compound as a 1: 1 diastereomeric mixture as a white foam, 33.2 mg, 11%. MS: 404 ES + (C ₁₆ H ₂₂ FN ₃ O ₈ ).

L NMR (300 MHz, DMSO-d6) δ: 1.23 (m, 6H); 1.46 (m, 3H); 1.62 (m, 3H); 3.05 (m, 1H); 3.79 (m, 1H); 4.00 (m, 1H); 4.19 (s, 1H); 4.80 (m, 1H); 6.00 (m, 1H); 6.33 (dd, 1H); 7.37 (s, 1H); 7.80 (d, 1H).

Example 54. (2R) -Benzyl-2 - (((2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy ) -2-fluoroacetate

DBU (8.83 ml, 58.57 mmol) in DMF (30 ml) was added dropwise to a solution of (2S, 5R) -6-hydroxy-3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct -3-ene-2-carboxamide (Intermediate 193, 10.5 g, 53.25 mmol) and (R) -benzyl-2-bromo-2-fluoroacetate (Intermediate 171, 14.47 g, 58.57 mmol) in DMF (100 ml) at -40 ° C for 30 min under nitrogen atmosphere. The resulting solution was stirred at -40 ° C for 30 min, then the reaction was stopped by adding water (15 ml). The reaction mixture was extracted with ethyl acetate (3x20 ml). The combined organics were washed with brine (3x20 ml), dried over sodium sulfate, filtered and concentrated. Chromatography on silica gel (0-40% ethyl acetate / petroleum ether) afforded the title compound as a white solid, 8.8 g, 45%. MS: 364 ES + (C ₁₇ H ₁₈ FN ₃ O ₅ ).

L NMR (400 MHz, DMSO-d6) δ: 1.62 (s, 3H); 2.92 (d, 1H); 3.75 (d, 1H); 3.99 (m, 1H); 4.02 (s, 1H); 5.25 (s, 2H); 5.76 (s, 1H); 6.33 (d, 1H); 7.38 (m, 5H); 7.39 (s, 1H); 7.85 (s, 1H).

Intermediate 77. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid

To a solution of methyl (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo 1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxylate (Intermediate 185, 538.5 mg, 1.65 mmol) in DCE (3 ml) in a 20 ml microwave tube was added trimethyltin hydroxide (477.17 mg, 2.64 mmol). The reaction was carried out in a microwave oven for 2 h at 80 ° C. The solvent was removed and the resulting residue was dissolved in ethyl acetate and washed three times with 0.01N HCl. potassium bisulfate solution and once with brine. The organics were dried over sodium sulfate, filtered and concentrated to give an orange foam, 649 mg, 100%. MS: 313 ES + (C ₁₄ H ₂₄ N ₂ O ₄ Si).

L NMR (300 MHz, DMSO-d6) δ: 0.00 (s, 6H); 0.78 (s, 9H); 1.50 (s, 3H); 2.92 (m, 1H); 3.47 (m, 1H); 3.53 (m, 1H); 3.77 (m, 1H); 5.85 (m, 1H).

Intermediate 78.2- [2 - [(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-ene -2-carbonyl] hydrazino] -2-oxoacetamide

To a solution of (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-ene-2-carboxylic acid (Intermediate 77, 397.5 mg, 1.02 mmol) in DMF (6 ml) at 0 ° C, oxamic acid hydrazide (209.83 mg, 2.04 mmol), HATU (387 mg, 1.02 mmol) and N , N-diisopropylethylamine (0.57 ml, 3.26 mmol). The reaction mixture was stirred for 1 hour, then diluted with ethyl acetate and washed once with saturated sodium bicarbonate solution. Water

- 71 037916 the solution contained part of the product and was subjected to a single extraction with ethyl acetate. The combined organics were dried over magnesium sulfate, filtered and concentrated to give a yellow oil. Chromatography on silica gel (0-5% methanol) afforded the title compound as a light yellow solid, 142 mg, 35%. MS: 398 ES + (C16H27N5O5Si).

1H NMR (300 MHz, DMSO-d6) δ: 0.00 (s, 6H); 0.78 (s, 9H); 1.53 (s, 3H); 2.95 (m, 1H); 3.60 (m, 1H); 3.67 (m, 1H); 4.04 (s, 1H); 5.98 (m, 1H); 7.75 (s, 1H); 8.07 (s, 1H); 10.30 (s, 1H); 10.48 (s, 1H).

Intermediate 79.5 - [(2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-en-2- yl] -1,3,4-oxadiazole-2-carboxamide o h ₂ n - /

To a solution of 2- [2 - [(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carbonyl] hydrazino] -2-oxo-acetamide (Intermediate 78, 142 mg, 0.36 mmol) in DCM (6 ml) 4-nitrobenzenesulfonyl chloride (79.17 mg, 0.36 mmol) was added at room temperature and N, N-diisopropylethylamine (0.19 ml, 1.07 mmol). The reaction mixture turned yellow and was stirred for 30 min, then diluted with dichloromethane and washed with brine. The organics were dried over magnesium sulfate, filtered and concentrated to give an orange oil. Chromatography on silica gel (0-5% methanol / dichloromethane) afforded the title compound as an orange foam, 93.8 mg, 69%. MS: 380 ES + (C16H25N5O4Si).

¹ H NMR (300 MHz, DMSO) δ: 0.00 (s, 6H); 0.77 (s, 9H); 1.53 (s, 3H); 3.02 (m, 2H); 3.70 (m, 1H); 5.01 (s, 1H); 6.14 (m, 1H); 8.13 (s, 1H); 8.53 (s, 1H).

Intermediate 80.5 - [(2S, 5R) -6-Hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-en-2-yl] -1,3,4- oxadiazole-2-carboxamide .o h ₂ n ~ # Ι ^ Ο I vA

D - N ₄ O OH

To a solution of 5 - [(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2- yl] -1,3,4-oxadiazole-2-carboxamide (Intermediate 79, 267.6 mg, 0.71 mmol) in ethyl acetate (6 ml) was added HF-pyridine (0.04 ml, 1.41 mmol) ... The reaction mixture was stirred for 2.5 h, then another 2 eq. HF-pyridine and the reaction mixture was stirred for another hour. The reaction mixture was concentrated to give the title compound as a brownish beige solid, 245 mg, 99%. MS: 266 ES + (C10H11N5O4).

1H NMR (300 MHz, DMSO-d6) δ: 1.52 (s, 3H); 3.01 (m, 2H); 3.66 (m, 1H); 4.95 (s, 1H); 6.18 (m, 1H); 8.09 (s, 1H); 8.45 (m, 1H).

Intermediate 81. Ethyl- (2S) -2 - [[(2S, 5R) -2- (5-carbamoyl-1,3,4-oxadiazol-2-yl) -3methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetate

To a solution of 5 - [(2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-yl] -1,3,4- oxadiazole-2-carboxamide (Intermediate 80, 176.8 mg, 0.67 mmol) in DMF (5 ml) was added ethyl (2S) -2-bromo-2-fluoroacetate (0.16 ml, 1.33 mmol ) and potassium carbonate (276.39 mg, 2 mmol). The reaction mixture was stirred for 2 h, then diluted with ethyl acetate, filtered and washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated to give a yellow oil. Chromatography on silica gel (0-70% ethyl acetate / hexanes) afforded the title compound as a light yellow film, 66.8 mg, 27%. The compound was a 7: 3 mixture of diastereomers. MS: 370 ES + (C14H16FN5O6).

1H NMR (300 MHz, DMSO-f) δ: 1.25 (m, 3H); 1.72 (s, 3H); 3.20 (m, 2H); 4.19 (m, 1H); 5.30 (m, 1H); 6.25 (m, 1H); 6.26 (m, 1H); 8.26 (s, 1H); 8.65 (s, 1H).

Example 55.2 - [[(2S, 5R) -2- (5-Carbamoyl-1,3,4-oxadiazol-2-yl) -3-methyl-7-oxo-1,6-diazabicyclo

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[3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetic acid lithium salt

To a solution of ethyl- (2S) -2 - [[(2S, 5R) -2- (5-carbamoyl-1,3,4-oxadiazol-2-yl) -3-methyl-7-oxo-1,6- diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetate (Intermediate 81, 66.8 mg, 0.18 mmol) in THF (1 ml) and water (0.5 ml ) at 0 ° C was added lithium hydroxide (0.18 ml, 0.18 mmol). The reaction mixture was stirred for 10 minutes. Added another 0.5 equivalents of lithium hydroxide. After 10 minutes, the reaction mixture was treated with an additional 0.5 equivalents of lithium hydroxide. The reaction mixture was stirred for 20 min, neutralized by the addition of 0.5 N HCl. HCl, frozen and lyophilized to give a yellow solid. Reverse phase ISCO (50 g RediSep Gold C18, 100% water, 4 min; then 0% -50% acetonitrile / water) gave the title compound as an off-white solid, 26 mg, 36%. The compound was a 7: 3 mixture of diastereomers. MS: 342 ES + (C ₁₂ H ₁₂ FN ₅ O ₆ ).

NMR (300 MHz, DMSO-d ₆ ) δ: 1.70 (s, 3H); 3.20 (m, 2H); 4.15 (m, 1H); 5.20 (s, 1H); 5.28 (m, 1H); 6.27 (m, 1H); 8.25 (s, 1H); 8.65 (s, 1H).

Intermediate 82.2- (1,3-Dioxoisoindolin-2-yl) oxyethanesulfonamide ^h ^ ⁿ ^ nA ό Λ / Λ o \ _ /

To a suspension of 2-hydroxyethanesulfonamide (Enamine, 1.92 ml, 7.19 mmol), N-hydroxyphthalimide (1.41 g, 8.63 mmol) and triphenylphosphine (2.26 g, 8.63 mmol) at 0 ° С diisopropyl azodicarboxylate (1.7 ml, 8.63 mmol) was added dropwise. The reaction mixture turned dark orange and then pale yellow in color. After stirring for about 3 hours, the reaction mixture was concentrated to give a viscous pale yellow oil, which was purified by washing with ethyl acetate / hexanes. The white solid was collected by filtration and was the title compound, 1.8 g, 70%. MS: 271 ES + (C10H10N2O5S) NMR (300 MHz, DMSO ^) δ: 3.52 (m, 2H); 4.50 (m, 2H); 6.96 (s, 2H); 7.87 (s, 4H).

Intermediate 83.2-Aminoxyethanesulfonamide ^H2N '| ^ O' _N h ₂ o

To a solution of 2- (1,3-dioxoisoindolin-2-yl) oxyethanesulfonamide (Intermediate 82, 740 mg, 2.05 mmol) in DCM (20 ml) was added methyl hydrazine (0.11 ml, 2.05 mmol ). A precipitate formed immediately. The suspension was stirred at room temperature for about 2 hours, then concentrated. The solid was purified by washing with DCM and collected by filtration. The solid was purified by washing with methanol and the resulting white solid was filtered off. The NMR spectrum of the solid indicated that it was a by-product. The filtrate was concentrated to give the title compound as an off-white solid, 228.2 mg, 58%. The NMR spectrum showed that 73% of the target product was obtained. NMR (300 MHz, DMSO-d ₆ ) δ: 3.27 (m, 2H); 3.85 (m, 2H); 6.11 (bs, 2H); 6.79 (s, 2H).

Intermediate 84. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N- (2sulfamoylethoxy) -1,6-diazabicyclo [3.2.1] oct-3 -ene-2-carboxamide

To a solution of (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-ene-2-carboxylic acid (Intermediate 77, 465.74 mg, 1.19 mmol) in DMF (8 ml) at 0 ° C was added 2-aminooxyethanesulfonamide (Intermediate 83, 228.96 mg, 1.19 mmol), HATU (453.43 mg, 1 , 19 mmol) and N, N-diisopropylethylamine (0.57 ml, 3.26 mmol). The reaction mixture was then stirred for 1 h at 0 ° C, then diluted with ethyl acetate and washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated to give a yellow oil. Chromatography on silica gel (0% 80% ethyl acetate / hexanes) afforded the title compound as a colorless oil, 343 mg, 66%. MS: 435 ES + (C ^^ OeSiS).

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1H NMR (300 MHz, DMSO-d6) δ: 0.00 (s, 6H); 0.78 (s, 9H); 1.45 (s, 3H); 2.97 (m, 1H); 3.20 (m, 2H); 3.57 (m, 2H); 3.84 (m, 1H); 4.02 (m, 2H); 6.01 (m, 1H); 6.80 (s, 2H); 11.74 (s, 1H).

Intermediate 85: (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N- (2-sulfamoylethoxy) -1,6diazabicyclo [3.2.1] oct-3-en-2-carboxamide

To a solution of (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N- (2-sulfamoylethoxy) -1.6 diazabicyclo [3.2.1] oct-3-ene -2-carboxamide (Intermediate 84, 343 mg, 0.79 mmol) in ethyl acetate (4 ml) was added HF-pyridine (0.04 ml, 1.58 mmol). The reaction mixture was stirred for 30 minutes. Added another 2 eq. HF-pyridine and the reaction mixture was stirred for another hour. After 6 hours and a total of 7 eq. The HF-pyridine reaction was complete. The reaction mixture was filtered to collect an off-white solid. The solid became viscous and adhered to the filter. Ethyl acetate and a small amount of methanol were used to rinse the filter and transfer the solids to the flask. The solvent was removed in vacuo to give the title compound as an off-white solid, 337 mg, 100%. MS: 321 ES + (C ₁₀ H ₁₆ N ₄ O 6 S).

Intermediate 86. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (2-sulfamoylethoxycarbamoyl) -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N- (2-sulfamoylethoxy) -1,6-diazabicyclo [3.2.1] oct-3-en-2- carboxamide (Intermediate 85, 279.32 mg, 0.65 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.08 ml, 0.65 mmol) according to the procedure for example 45 to give a light yellow oil, 26.7 mg, 10%. MS: 425 ES + (C14H21FN4O8S).

1H NMR (300 MHz, DMSO-dg) δ: 1.25 (m, 3H); 1.60 (m, 3H); 3.15 (m, 1H); 3.36 (m, 1H); 3.78 (m, 1H); 4.02 (m, 3H); 4.07 (m, 2H); 4.28 (m, 2H); 6.13 (m, 1H); 6.16 (d, 1H); 6.95 (s, 2H) 11.91 (s, 1H).

Example 56 (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (2-sulfamoylethoxycarbamoyl) -1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetic acid lithium salt

To a solution of ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (2-sulfamoylethoxy-carbamoyl) -1.6 diazabicyclo [3.2.1] oct- 3-en-6-yl] oxy] acetate (Intermediate 86, 26.7 mg, 0.06 mmol) in THF (1 ml) and water (0.5 ml), LiOH (0.06 ml, 0.06 mmol). The reaction mixture was stirred for 15 minutes and another 0.5 equivalent of lithium hydroxide was added. After 15 minutes, another 0.5 eq. lithium hydroxide was added. After 30 minutes and after slight heating, the reaction was complete. The reaction mixture was neutralized by the addition of 0.5 N. HCl, frozen and lyophilized to give a yellow oil. The compound was purified by reverse phase ISCO (5.5 rRediSep Gold C18, 100% water). The title compound was obtained as an off-white solid, 11.3 mg, 29%. MS: 397 ES + (C ₁₂ H ₁₇ FN ₄ O ₈ S).

1H NMR (300 MHz, DMSO-dg) δ: 1.56 (s, 3H); 3.02 (m, 1H); 3.26 (m, 1H); 3.98 (m, 5H); 5.20 (d, 1H); 6.03 (m, 1H); 7.06 (s, 2H).

Intermediate 87. (2S, 5R) -6-Allyloxy-3-methyl-7-oxo-N- (2-sulfamoylethyl) -1,6diazabicyclo [3.2.1] oct-3-en-2-carboxamide

The title compound was prepared from (2S, 5R) -6-allyloxy-3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 29, 208.5 mg, 0.88 mmol) and 2-amino-ethylsulfonamide hydrochloride (281.1 mg, 1.75 mmol) according to the procedure for Intermediate 84 to give a pale yellow oil, 86.6, 29%. MS: 345 ES + (C ₁₃ H ₂₀ N ₄ O ₅ S).

- 74 037916

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.63 (s, 3H); 3.04 (m, 1H); 3.14 (m, 2H); 3.26 (m, 1H); 3.51 (m, 2H);

3.94 (m, 1H); 4.09 (m, 1H); 4.36 (m, 2H); 5.27 (m, 2H); 5.95 (m, 1H); 6.07 (m, 1H); 6.89 (s, 2H); 8.52 (m, 1H).

Intermediate 88. (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N- (2-sulfamoylethyl) -1,6diazabicyclo [3.2.1] oct-3-en-2-carboxamide

To a solution of (2S, 5R) -6-αlyloxy-3-methyl-7-oxo-N- (2-sulfamoylethyl) -1,6-diazabicyclo [3.2.1] oct3-en-2-carboxamide (Intermediate 87, 86.6 mg, 0.25 mmol) in methanol (3 ml), 1,3-dimethylbarbituric acid (78.53 mg, 0.5 mmol) and tetrakis (triphenylphosphine) palladium (0) (58.12 mg, 0.05 mmol). The reaction mixture was stirred for 1 hour at room temperature, then concentrated to give a dark orange oil, 76.5 mg, 100%. MS: 305 ES + (C10H16N4O5S).

Intermediate 89. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (2-sulfamoylethylcarbamoyl) -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate

To a solution of (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N- (2-sulfamoylethyl) -1,6-diazabicyclo [3.2.1] oct-3ene-2-carboxamide (Intermediate 88, 76.5 mg, 0.25 mmol) in 1,4-dioxane (2 ml) and DMF (0.25 ml) was added ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.09 ml, 0.75 mmol). The reaction mixture was cooled to 0 ° C and DBU (0.04 mL, 0.25 mmol) was added dropwise. Ethyl (2S) -2-bromo-2-fluoroacetate (0.09 ml, 0.75 mmol) was added again. Then DBU (0.04 ml, 0.25 mmol) was added again. After another 15 min at 0 ° C with periodic heating, another 0.5 eq. DBU. The reaction mixture was stirred for an additional 15 minutes at room temperature. The reaction mixture was diluted with ethyl acetate and washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated to give an orange oil. Chromatography on silica gel (0-25% acetone / dichloromethane) afforded the title compound with some triphenylphosphine oxide as an orange film, 77.3 mg, 75%. MS: 409 ES + (C14H21FN4O7S).

Example 57 (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (2-sulfamoylethylcarbamoyl) -1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetic acid lithium salt

To a solution of ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (2-sulfamoylethyl-carbamoyl) -1.6 diazabicyclo [3.2.1] oct- 3-en-6-yl] oxy] acetate (Intermediate 89, 72.3 mg, 0.18 mmol) in THF (2 ml) and water (1 ml), 1M lithium hydroxide (0.18 ml, 0.18 mmol). The reaction mixture was stirred for 10 min at 0 ° C, then neutralized by the addition of 0.5 N. hydrochloric acid, frozen and lyophilized to give a yellow solid. Reverse phase ISCO (100% water) gave the title compound as a light yellow solid, 14.3 mg, 21%. MS: 381 ES + (C ₁₂ H ₁₇ FN ₄ O ₇ S).

H NMR (300 MHz, DMSO-46) δ: 1.64 (s, 3H); 3.13 (m, 3H); 3.52 (m, 3H); 3.99 (m, 1H); 4.14 (m, 1H); 5.24 (d, 1H); 6.04 (m, 1H); 6.90 (m, 2H); 8.57 (m, 1H).

Example 58. Ethyl-2 - [[(2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en6 -yl] oxy] -2-fluoroacetyl] oxymethoxycarbonylamino] -3-methylbutanoate

To a solution of (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetic acid (example 4, 100 mg, 0.37 mmol), Hunig's base (0.06 ml, 0.37 mmol) and ethyl 2- (chloromethoxycarbonylamino) -3-methyl-butanoate (0.05 ml , 0.73 mmol) in DMF (1.5 ml) was added tetrabutyl ammonium chloride (83.42 mg, 0.37 mmol) at room temperature. Reaction mixture

- 75 037916 was stirred at 40 ° C for 2 hours. Then it was diluted with ethyl acetate and washed twice with brine / water (1: 1). The organics were dried over anhydrous magnesium sulfate, filtered and concentrated to give an orange oil. Chromatography on silica gel (0-80% ethyl acetate / hexanes) afforded the title compound (7 mg, 3.63%) as a white solid. MS: 475 ES + (C19H27FN4O9).

1H NMR (300 MHz, DMSO-d6) δ: 0.90 (m, 6H); 1.21 (m, 3H); 1.61 (s, 3H); 2.15 (m, 1H); 3.78 (m, 1H); 3.95 (m, 1H); 4.01 (m, 1H); 4.15 (m, 2H); 4.22 (m, 1H); 5.80 (m, 2H); 5.95 (m, 1H); 6.30 (d, 1H); 7.40 (s, 1H); 7.80 (s, 1H); 8.10 (d, 1H).

Intermediate 90. tert-Butyl- (2S) -2 - [(1,3-dioxoisoindolin-2-yl) oxymethyl] pyrrolidine-1-carboxylate

To a solution of diethyl azodicarboxylate (14.21 ml, 12.48 mmol) in THF (10 ml) at -10 ° C was added a solution of triphenylphosphine (3273.22 mg, 12.48 mmol) in THF (20 ml) dropwise. The suspension was stirred at -10 ° C. After 20 minutes, the suspension turned solid and another 40 ml of THF was added. After 1 h, a solution of Boc-L-prolinol (448.18 ml, 5.94 mmol) in THF (10 ml) was added, followed by a solution of N-hydroxyphthalimide (969.41 mg, 5.94 mmol) in THF (10 ml). The reaction mixture was allowed to warm to room temperature and was stirred overnight. It was concentrated and the resulting oil was purified by washing with ethyl acetate / hexanes. The residue was removed by filtration and the filtrate was concentrated on silica gel. Chromatography on silica gel (0% -40% EtOAc / Hexanes) afforded the title compound (2.37 g, quant.) As a light yellow solid. MS: 347 ES + (C18H22N2O5).

Intermediate 91. tert-Butyl- (2S) -2- (aminooxymethyl) pyrrolidine-1-carboxylate

To a solution of tert-butyl (2S) -2 - [(1,3-dioxoisoindolin-2-yl) oxymethyl] pyrrolidine-1-carboxylate (Intermediate 90, 2.06 g, 5.95 mmol) in DCM (20 ml) hydrazine monohydrate (2.14 ml, 17.84 mmol) was added at room temperature. A white precipitate formed immediately. The suspension was stirred at room temperature for 1 hour, then filtered through celite. The filtrate was washed twice with brine / water (1: 1), dried over anhydrous magnesium sulfate, filtered and concentrated to give the title compound (1.35 g, quant.) As a viscous oil.

¹ H NMR (300 MHz CDCla) δ: 1.50 (s, 9H); 1.85 (m, 4H); 3.32 (m, 2H); 3.63 (m, 1H); 4.25 (m, 2H).

Intermediate 92. tert-Butyl- (2S) -2 - [[[(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3methyl-7-oxo-1,6-diazabicyclo [3.2. 1] oct-3-ene-2-carbonyl] amino] oxymethyl] pyrrolidine-1-carboxylate

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) siluyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid acid (Intermediate 77, 1.85 g, 5.92 mmol) and tert-butyl- (2S) -2- (aminooxymethyl) pyrrolidine-1-carboxylate (Intermediate 91, 1.28 g, 5.92 mmol) according to the procedure for intermediate 84 to give (585 mg, 19%) as a white tacky foam. MS: 511 ES + (C ₂₄ H ₄₂ N ₄ O ₆ Si).

Intermediate 93. tert-Butyl- (2S) -2 - [[[(2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carbonyl] amino] oxymethyl] pyrrolidine-1-carboxylate

To a solution of tert-butyl- (2S) -2 - [[[(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carbonyl] amino] oxymethyl] pyrrolidine-1-carboxylate (Intermediate 92, 401 mg, 0.79 mmol) in ethyl acetate (20 ml) at 0 ° C under nitrogen atmosphere, HF pyridine (24 μl , 0.94 mmol). The reaction mixture was stirred at room temperature for

- 76,037916 h, then concentrated. The crude material was partitioned between DCM (100 ml) and brine (50 ml).

The organic layer was dried over anhydrous sodium sulfate and concentrated to give the title compound (309 mg, 84%) as a white sticky solid. MS: 396 ES + (C18H28N4O6).

Example 59 tert-Butyl- (2S) -2 - ((((2S, 5R) -6 - ((S) -2-ethoxy-1-fluoro-2-oxoethoxy) -3-methyl-7-oxo 1,6 diazabicyclo [3.2.1] oct-3-ene-2-carboxamido) oxy) methyl) pyrrolidine-1-carboxylate

To a suspension of tert-butyl- (2S) -2 - [[[(2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-2-carbonyl ] amino] oxymethyl] pyrrolidine-1-carboxylate (Intermediate 93, 309 mg, 0.78 mmol) and cesium carbonate (304.75 mg, 0.94 mmol) in THF (15 ml), ethyl (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.14 mL, 1.17 mmol). The reaction mixture was stirred at 0 ° C for 8 h. Water (50 ml) and EtOAc (100 ml) were added. The organic layer was dried over anhydrous sodium sulfate, concentrated, and purified by flash chromatography (20 g silica gel, 0-100% EtOAc / hexanes) to give the title compound (164 mg, 40%) as a sticky solid. MS: 501 ES + (C ₂₂ H ₃₃ FN ₄ O ₈ ).

¹ H NMR (300 MHz, CDCl ₃ ) δ: 1.38 (m, 3H); 1.45 (s, 9H); 1.80 (s, 3H); 1.95 (m, 4H); 3.35 (m, 3H); 3.60 (m, 2H); 3.95 (m, 1H); 4.01 (m, 1H); 4.28 (m, 2H); 4.35 (m, 2H); 5.75 (d, 1H); 6.10 (s, H).

Example 60. Ethyl- (2S) -2-fluoro-2 - (((2S, 5R) -3-methyl-7-oxo-2 - ((((S) -pyrrolidin-2-yl) methoxy) carbamoyl) -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) acetate TFA salt

To a solution of tert-butyl- (2S) -2 - ((((2S, 5R) -6 - ((S) -2-ethoxy-1-fluoro-2-oxoethoxy) -3-methyl-7-oxo-1 , 6diazabicyclo [3.2.1] oct-3-ene-2-carboxamido) oxy) methyl) pyrrolidine-1-carboxylate (example 59, 76 mg, 0.15 mmol) in DCM (10 ml) at 0 ° С dropwise TFA (0.58 ml, 7.59 mmol) was added. The reaction mixture was stirred at 0 ° C for 2 h, then concentrated. The residue was purified by washing with diethyl ether to give the title compound (53 mg, 83%) as a TFA salt. MS: 401 ES + (C17H25FN4O6).

1 H NMR (300 MHz, CDCl ₃ ) δ: 1.45 (m, 3H); 1.86 (m, 4H); 2.15 (m, 3H); 3.55 (m, 4H); 4.01 (m, 2H); 4.32 (m, 5H); 5.85 (d, 1H); 6.13 (s, 1H).

Intermediate 94. tert-Butyl- (S) -2 - (((1,3-dioxoisoindolin-2-yl) oxy) methyl) -4,4 difluoropyrrolidine-1-carboxylate o

f F Α-ΆΧ

AxA'VU

About about% /

The title compound was prepared from tert-butyl (S) -4,4-difluoro-2- (hydroxymethyl) pyrrolidine-1-carboxylate (2.8 g, 11.8 mmol) following the procedure for Intermediate 90 to give ( 4.3 g, 95%) of a pale yellow sticky solid. MS: 383 ES + (C ₁₈ H ₂₀ F ₂ N ₂ O ₅ ).

Intermediate 95: tert-butyl- (S) -2 - ((aminooxy) methyl) -4,4-difluoropyrrolidine-1carboxylate

The title compound was prepared from tert-butyl (S) -2 - (((1,3-dioxoisoindolin-2yl) oxy) methyl) -4,4-difluoropyrrolidine-1-carboxylate (Intermediate 94, 1.9 g , 4.97 mmol) according to the procedure for Intermediate 91 to give (1.25 g, 99%) as an orange viscous oil.

1 H NMR (300 MHz, CDCl ₃ ) δ: 1.51 (s, 9H); 2.42 (m, 2H); 3.63 (m, 2H); 3.85 (m, 2H); 4.97 (m, 1H).

- 77 037916

Intermediate 96. tert-Butyl- (2S) -2 - ((((2S, 5R) -6 - ((tert-butyldimethylsilyl) oxy) -3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamido) oxy) methyl) -4,4-difluoropyrrolidine-1 carboxylate

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 1.3 g, 4.16 mmol) and tert-butyl- (S) -2 - ((aminooxy) methyl) -4,4-difluoropyrrolidine-1-carboxylate (Intermediate 95, 1, 26 g, 4.99 mmol) according to the procedure for Intermediate 84 to give (489 mg, 18%) as a sticky white foam. MS: 547 ES + (C ₂₄ H ₄₀ F ₂ N ₄ O ₆ Si).

Intermediate 97. tert-Butyl- (2S) -4,4-difluoro-2 - ((((2S, 5R) -6-hydroxy-3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct -3-en-2-carboxamido) oxy) methyl) pyrrolidine-1 -carboxylate

The title compound was prepared from tert-butyl- (2S) -2 - ((((2S, 5R) -6 - ((tert-butyldimethylsilyl) oxy) -3-methyl-7-oxo-1,6-diazabicyclo [ 3.2.1] oct-3-ene-2-carboxamido) oxy) methyl) -4,4 difluoropyrrolidine-1-carboxylate (Intermediate 96, 489 mg, 0.89 mmol) according to the procedure for Intermediate 93 to give (330 mg , 72%) as a white sticky solid. MS: 433 ES + (C18H26F2N4O6).

Intermediate 98.tert-Butyl- (2S) -2 - ((((2S, 5R) -6 - ((S) -2-ethoxy-1-fluoro-2-oxoethoxy) 3-methyl-7-oxo 1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamido) oxy) methyl) -4,4-difluoropyrrolidine-1carboxylate

To a suspension of tert-butyl- (2S) -4,4-difluoro-2 - ((((2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct -3-ene-2-carboxamido) oxy) methyl) pyrrolidine-1-carboxylate (Intermediate 97, 300 mg, 0.69 mmol) and cesium carbonate (339 mg, 1.04 mmol) in ethyl acetate (15 ml) at Ethyl (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.1 mL, 0.83 mmol) was added at 0 ° C. The reaction mixture was stirred at 10 ° C for 1 hour. Water (50 ml) and EtOAc (100 ml) were added. The organic layer was separated, concentrated and purified by flash chromatography on silica gel (0-100%, EtOAc / hexane) to give the title compound (70 mg, 18%) as a sticky white foam. MS: 537 ES + (C22H ₃ 'F ₃ N4O ₈ ).

Example 61. Ethyl- (2S) -2 - ((((2S, 5R) -2 - ((((S) -4,4-difluoropyrrolidin-2-yl) methoxy) carbamoyl) -3methyl-7-oxo 1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetate TFA salt

The title compound was prepared from tert-butyl- (2S) -2 - ((((2S, 5R) -6 - ((S) -2-ethoxy-1-fluoro-2-oxoethoxy) -3-methyl-7-oxo -1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamido) oxy) methyl) -4,4-difluoropyrrolidine-1-carboxylate (Intermediate 98, 26 mg, 0.05 mmol) according to the procedure for example 60 to obtain (20 mg, 85%) as a TFA salt. MS: 437 ES + (C ₁₇ H ₂₃ F ₃ N ₄ O ₆ ).

1H NMR (300 MHz, CDCl3) δ: 1.42 (m, 3H); 1.80 (s, 3H); 2.65 (m, 2H); 3.45 (m, 2H); 3.82 (m, 2H); 4.10 (m, 1H); 4.38 (m, 6H); 5.80 (d, 1H); 6.18 (s, 1H).

Intermediate 99. (2S) -2 - [[(2S, 5R) -2 - [[(2S) -1-tert-butoxycarbonylpyrrolidin-2-yl] methoxycarbamoyl] -3-methyl-7-oxo-1,6 -diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetic acid

- 78 037916

To a solution of tert-butyl- (2S) -2 - ((((2S, 5R) -6 - ((S) -2-ethoxy-1-fluoro-2-oxoethoxy) -3-methyl-7-oxo-1 , 6diazabicyclo [3.2.1] oct-3-ene-2-carboxamido) oxy) methyl) pyrrolidine-1-carboxylate (example 59, 81 mg, 0.16 mmol) in THF (1 ml) and water (0.50 ml) at 0 ° C was added lithium hydroxide (1N, 0.01 ml, 0.40 mmol). The reaction mixture was stirred for 30 minutes. Dilute HCl solution (0.5N) was added to adjust the pH to 2. The reaction mixture was extracted with EtOAc (20 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated to give the title compound (50 mg, 65%) as a gum. MS: 471 ES- (C ₂₀ H ₂₉ FN ₄ O ₈ ).

Example 62. (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [[(2S) -pyrrolidin-2-yl] methoxycarbamoyl] 1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetic acid TFA salt

The title compound was prepared from (2S) -2 - [[(2S, 5R) -2 - [[(2S) -1-tert-butoxycarbonylpyrrolidin-2-yl] methoxycarbamoyl] -3-methyl-7-oxo-1 , 6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2fluoroacetic acid (Intermediate 99, 65 mg, 0.14 mmol) according to the procedure for Example 60 to obtain (40 mg, 70% ) as a TFA salt. MS: 473 ES + (C ₂₀ H ₂₉ FN ₄ O ₈ ).

¹ H NMR (300 MHz, D2 O) δ: 1.65 (s, 3H); 1.82 (m, 1H); 2.23 (m, 2H); 2.35 (m, 1H); 3.40 (m, 3H); 3.58 (m, 1H); 3.82 (m, 1H); 4.02 (m, 1H); 4.18 (m, 1H); 4.33 (m, 2H); 5.70 (d, 1H); 6.28 (s, 1H).

Intermediate 100. (2S) -2 - (((2S, 5R) -2 - ((((S) -1- (tert-Butoxycarbonyl) -4,4-difluoropyrrolidin-2-yl) methoxy) carbamoyl) - 3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2fluoroacetic acid

The title compound was prepared from tert-butyl- (2S) -2 - ((((2S, 5R) -6 - ((S) -2-ethoxy-1-fluoro-2-oxoethoxy) -3 -methyl-7-oxo -1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxamido) oxy) methyl) -4,4-difluoropyrrolidine-1-carboxylate (Intermediate 98, 37 mg, 0.07 mmol) according to the procedure for Intermediate 99 to give (35 mg, 90%) as a gum. MC: 507 ES- (C ₂₀ H ₂₇ F ₃ N ₄ O ₈ ).

Example 63. (2S) -2 - (((2S, 5R) -2 - ((((S) -4,4-Difluoropyrrolidin-2-yl) methoxy) carbamoyl) -3-methyl-7oxo-1,6 -diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetic acid TFA salt

The title compound was prepared from (2S) -2 - (((2S, 5R) -2 - ((((S) -1- (tert-butoxycarbonyl) 4,4-difluoropyrrolidin-2-yl) methoxy) carbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetic acid (Intermediate 100, 35 mg, 0.07 mmol) according to following the procedure for Example 60 to give (22 mg, 70%) as a TFA salt. MS: 409 ES + (C ₁₅ H ₁₉ F ₃ N ₄ O ₆ ).

1H NMR (300 MHz, D2O) δ: 1.78 (s, 3H); 2.62 (m, 1H); 2.95 (m, 1H); 3.13 (m, 1H); 3.42 (m, 1H); 3.58 (m, 1H); 3.96 (m, 2H); 4.61 (m, 1H); 4.40 (m, 3H); 5.82 (d, 1H); 6.33 (s, 1H).

Example 64 [(2R) -2 - [[(2S, 5R) -2-Carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] hydroxy] -2-fluoroacetyl] oxymethyl-2,2-dimethylpropanoate

- 79 037916

The title compound was prepared from (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6-yl] hydroxy] -2-fluoroacetic acid (example 4, 47 mg, 0.17 mmol) and chloromethyl pivalate (0.05 ml, 0.34 mmol) according to the procedure for example 53 to obtain (33.2 mg, 49.8%) as a white solid. MS: 388 ES + (C ₁₆ H ₂₂ FN ₃ O ₇ ).

'H NMR (300 MHz, DMSO-d6) δ: 1.12 (s, 9H); 1.62 (s, 3H); 3.01 (m, 1H); 3.82 (m, 1H); 4.02 (m, 1H); 4.19 (m, 1H); 5.76 (m, 1H); 5.87 (m, 1H); 5.91 (m, 1H); 6.25-6.43 (d, 1H); 7.41 (bs, 1H); 7.86 (bs, lH).

Example 65. Indan-5-yl- (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6 -yl] oxy] -2-fluoroacetate

To a solution of (2R) -2 - [[(2S, 5R) -2-carbamoyl-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetic acid (example 4, 100 mg, 0.37 mmol) and 5-indanol (58.9 mg, 0.44 mmol) in DCM (3 ml) and THF (3 ml) at 0 ° C was added Ν , '-Dicyclohexylcarbodiimide (113.3 mg, 0.55 mmol) and DMAP (5.0 mg). The reaction mixture was stirred at room temperature. pace. for 1 h. The reaction mixture was concentrated, then dissolved in EtOAC. The resulting white solid was filtered off. The filtrate was concentrated. After chromatography on silica gel (0-80% ethyl acetate / hexanes), the title compound (72 mg, 48%) was obtained as a white solid. MS: 390 ES + (C19H20FN3OX

1H NMR (300 MHz, DMSO-d6) δ: 1.64 (s, 3H); 2.05 (m, 2H); 2.85 (m, 4H); 3.10 (m, 1H); 3.79 (m, 1H); 4.06 (m, 1H); 4.22 (m, 1H); 6.06 (m, 1H); 6.44-6.51 (d, 1H); 6.86 (m, 1H); 6.97 (m, 1H); 7.28 (m, 1H); 7.43 (bs, 1H); 7.87 (bs, 1H).

Intermediate 101. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-N- (oxetane-3yl) -7-oxo 1,6-diazabicyclo [3.2.1] oct -3-en-2-carboxamide

To a solution of (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-ene-2-carboxylic acid (Intermediate 77, 100 mg, 0.32 mmol) in DMF (1.5 ml) at 0 ° C were added HATU (35 mg, 0.48 mmol) and Ν, Ν'-diisopropylethylamine (0.167 ml, 0.96 mmol). The reaction mixture was stirred for 30 min at room temperature, then diluted with ethyl acetate and washed once with saturated sodium bicarbonate solution and three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-2.5% methanol / dichloromethane) gave the title compound (80 mg, 68%) as a yellow oil. MS: 368 ES + (C ₁₇ H ₂₉ N ₃ O ₄ Si).

Intermediate 102. (2S, 5R) -6-Hydroxy-3-methyl-N- (oxetan-3-yl) -7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-2-carboxamide

(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3methyl-N- (oxetan-3-yl) -7-oxo-1,6-diazabicyclo [3.2. 1] oct-3-ene-2-carboxamide (Intermediate 101, 0.10 g, 0.22 mmol) in ethyl acetate (0.5 ml) at room temperature under nitrogen atmosphere. HF-pyridine (0.015 ml, 0.33 mmol) was added and the reaction mixture was stirred at room temperature for 1 h. The solvent was removed. MS: 254 ES + (C _n H ₁₅ N ₃ O ₄ ).

Intermediate 103. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-2- (oxetan-3-ylcarbamoyl) 7-oxo-1,6-diazabicyclo [3.2.1 ] oct-3-en-6-yl] oxy] acetate

To a solution of (2S, 5R) -6-hydroxy-3-methyl-N- (oxetane-3-yl) -7-oxo 1,6-diazabicyclo [3.2.1] oct-3-en-2carboxamide (Intermediate 102, 50 mg, 0.20 mmol) in 1,4-dioxane (2 ml) and DMF (0.25 ml) was added ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.047 ml, 0.39 mmol). The reaction mixture was cooled to 0 ° C and DBU (0.089 mL, 0.59 mmol) was added dropwise. The reaction mixture was stirred at 0 ° C for 15 min, then diluted with ethyl acetate and washed three times

- 80 037916 with a mixture of brine and water in a 1: 1 ratio. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-80% EtOAc / Hexane) afforded the title compound (58 mg, 82.2%) as a colorless oil. The compound was a mixture of diastereomers in a 2: 8 ratio. MS: 358 ES + (C ₁₅ H ₂₀ FN ₃ O ₆ ).

Example 66 (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-2- (oxetan-3-ylcarbamoyl) -7-oxo-1,6-diazabicyclo

[3.2.1] oct-3-en-6-yl] oxy] acetic acid lithium salt

To a solution of ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-2- (oxetan-3-ylcarbamoyl) -7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 103, 56.3 mg, 0.16 mmol) in THF (1.0 ml) and water (0.5 ml) hydroxide was added at 0 ° C lithium (1M) (0.50 ml, 0.50 mmol). The reaction mixture was stirred for 1 hour. HCl solution (1N) was added to adjust the pH to about 5-6. The solvent was removed. Elution with water (0% -5% ACN / water) on a sepabead column (saturated water first, then ACN, then water rinsed) and lyophilization gave the title compound (10 mg, 17.3%) as a white solid. MS: 330 ES + (C ₁₃ H ₁₆ FN ₃ O ₆ ).

¹ H NMR (300 MHz, D ₂ O) δ: 1.63 (s, 3H); 3.22 (m, 1H); 3.37 (m, 1H); 4.04 (m, 1H); 4.30 (s, 1H); 4.58 (m, 2H); 4.86 (m, 3H); 5.15-5.56 (d, 1H); 6.15 (m, 1H).

Intermediate 104. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-N- [2- (methanesulfonamido) ethyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2. 1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 300 mg, 0.96 mmol) in DMF (1.5 ml) and N- (2-aminoethyl) -methanesulfonamide hydrochloride (251.5 mg, 1.44 mmol) according to the procedure for Intermediate 101 to give (220 mg, 53%) as a white solid. MS: 433 ES + (C ₁₇ H ₃₂ N ₄ O ₅ SiS).

Intermediate 105. (2S, 5R) -6-Hydroxy-N- [2- (methanesulfonamido) ethyl] -3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-N- [2 (methanesulfonamido) ethyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2 .1] oct-3-ene-2-carboxamide (Intermediate 104, 0.12 g, 0.28 mmol) according to the procedure for Intermediate 102 to give a brownish-beige residue. MS: 319 ES + (C _n H ₁₈ N ₄ O ₅ S).

Intermediate 106. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- [2- (methanesulfonamido) ethylcarbamoyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2 .1] oct-3-en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -6-hydroxy-N- [2- (methanesulfonamido) ethyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carboxamide (Intermediate 105, 80 mg, 0.25 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.089 mL, 0.75 mmol) according to the procedure for Intermediate 103 to give (25 mg, 18.8%) a white solid. The compound was a mixture of diastereomers in a 1: 4 ratio. MS: 423 ES + (Ci ₅ H2 ₃ FN4O7S).

Example 67. (2S) -2-Fluoro-2 - [[(2S, 5R) -2- [2- (methanesulfonamido) ethylcarbamoyl] -3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct- 3-en-6-yl] oxy] acetic acid lithium salt

- 81 037916

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-2- (3methylsulfonylpropylcarbamoyl) -7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 106, 25 mg, 0.06 mmol) following the procedure for Example 66 to give (4.0 mg,

15.4%) after lyophilization of a white solid. MS: 395 ES + (C ₁₃ H ₁₉ FN ₄ O ₇ S).

1H NMR (300 MHz, D ₂ O) δ: 1.63 (s, 3H); 2.95 (s, 3H); 3.18-3.38 (m, 6H); 4.02 (m, 1H); 4.27 (s, 1H); 5.56-5.74 (d, 1H); 6.13 (m, 1H).

Intermediate 107. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-N- (oxazole-2ylmethyl) -7-oxo-1,6-diazabicyclo [3.2.1] oct -3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 400 mg, 1.28 mmol) and oxazol-2-yl-methylamine hydrochloride (258.4 mg, 1.92 mmol) according to the procedure for Intermediate 101 to give (120 mg, 23.9% ) a white solid. MS: 393 ES + (C18H28N4O4Si).

Intermediate 108. (2S, 5R) -6-Hydroxy-3-methyl-N- (oxazol-2-ylmethyl) -7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methylN- (oxazol-2-ylmethyl) -7-oxo-1,6-diazabicyclo [3.2.1 ] oct-3-ene-2-carboxamide (Intermediate 107, 0.12 g, 0.31 mmol) according to the procedure for Intermediate 102 to give a residue. MS: 279 ES + (C12H14N4O4).

Intermediate 109. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-2- (oxazol-2-ylmethylcarbamoyl) -7-oxo-1,6-diazabicyclo [3.2. 1] oct-3-en-6-yl] oxy] acetate

O

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-N- (oxazol-2-ylmethyl) 7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2 -carboxamide (Intermediate 108, 80 mg, 0.29 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.10 mL, 0.86 mmol) according to the procedure for Intermediate 103 to give (15 mg, 13.6%) a colorless oil. The compound was a mixture of diastereomers in the ratio 15:85. MS: 383 ES + (C ₁₆ H ₁₉ FN ₄ O ₆ ).

Example 68. (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-2- (oxazol-2-ylmethylcarbamoyl) -7-oxo-1,6-diazabicyclo [3.2.1] oct -3-en-6-yl] oxy] acetic acid lithium salt

O

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-2- (oxazole-2ylmethylcarbamoyl) -7-oxo-1,6-diazabicyclo [3.2. 1] oct-3-en-6-yl] oxy] acetate (Intermediate 109, 15 mg, 0.039 mmol) according to the procedure for Example 66 to give (4.0 mg, 25.9%) after lyophilization of a white solid. MS: 355 ES + (C ₁₄ H ₁₅ FN ₄ O ₆ ).

L NMR (300 MHz, D2O) δ: 1.62 (s, 3H); 3.28 (m, 2H); 4.03 (m, 1H); 4.36 (s, 1H); 4.47 (m, 2H); 5.565.74 (d, 1H); 6.13 (m, 1H); 7.01 (d, 1H); 7.71 (d, 1H).

Intermediate 110. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N (pyrazin-2-ylmethyl) -1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 200 mg, 0.64 mmol) and pyrazin-2-yl-methylamine oxalate (191.2 mg, 0.96 mmol) according to the procedure for Pro- 82 037916 liquid compound 101 to give (120 mg, 46.4%) of a white solid. MS: 404 ES + (C19H29N5O ₃ Si).

Intermediate 111. (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N- (pyrazin-2-ylmethyl) -1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-N- (pyrazin-2-ylmethyl) -1,6-diazabicyclo [3.2.1 ] oct-3-ene-2-carboxamide (Intermediate 110, 0.10 g, 0.26 mmol) according to the procedure for Intermediate 102 to give a residue. MS: 290 ES + (C13H15N5O3).

Intermediate 112. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (pyrazin-2-ylmethylcarbamoyl) -1,6-diazabicyclo [3.2. 1] oct-3-en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N- (pyrazine-2ylmethyl) -1,6-diazabicyclo [3.2.1] oct-3-ene-2- carboxamide (Intermediate 111, 70 mg, 0.24 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.086 ml, 0.73 mmol) according to the procedure for Intermediate 103 to give ( 50 mg, 52.5%) of a colorless oil. The compound was a mixture of diastereomers in the ratio 15:85. MS: 394 ES + (C17H20FN5O5).

Example 69. (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (pyrazin-2-ylmethylcarbamoyl) -1,6-diazabicyclo [3.2.1] oct -3-en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 (pyrazin-2-ylmethylcarbamoyl) -1,6-diazabicyclo [3.2 .1] oct-3-en-6-yl] oxy] acetate (Intermediate 112, 50 mg, 0.13 mmol) according to the procedure for Example 66 to obtain (4.0 mg, 8.2%) after lyophilization of white solid matter. MS: 366 ES + (C ₁₅ H ₁₆ FN ₅ O ₅ ).

1H NMR (300 MHz, D2O) δ: 1.62 (s, 3H); 3.28 (m, 2H); 4.03 (m, 1H); 4.36 (s, 1H); 4.53 (m, 2H); 5.565.75 (d, 1H); 6.13 (m, 1H); 8.49 (m, 3H).

Intermediate 113. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-N- (cyclopropylmethoxy) -3 -methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 200 mg, 0.64 mmol) and O- (cyclopropylmethyl) hydroxylamine (83.6 mg, 0.96 mmol) according to the procedure for Intermediate 101 to give (100 mg, 40.9%) a colorless oils. MS: 382 ES + (C18H31N3O4Si).

Intermediate 114. (2S, 5R) -N- (Cyclopropylmethoxy) -6-hydroxy-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-N (cyclopropylmethoxy) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct- 3-ene-2-carboxamide (Intermediate 113, 0.1 g, 0.26 mmol) according to the procedure for Intermediate 102 to give a residue. MS: 268 ES + (C12H17N3O4).

Intermediate 115. Ethyl- (2S) -2 - [[(2S, 5R) -2- (cyclopropylmethoxycarbamoyl) -3-methyl7-oxo-1, 6-diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoro acetate

- 83 037916

O

The title compound was prepared from (2S, 5R) -N- (cyclopropylmethoxy) -6-hydroxy-3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 114, 60 mg, 0.22 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.08 mL, 0.67 mmol) according to the procedure for Intermediate 103 to give (15 mg , 18%) of a colorless oil. The compound was a mixture of diastereomers in a 1: 9 ratio. MS: 372 ES + (C ₁₆ H ₂₂ FN ₃ O ₆ ).

Example 70. (2S) -2 - [[(2S, 5R) -2- (Cyclopropylmethoxycarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl ] hydroxy] -2-fluoroacetic acid lithium salt

O

The title compound was prepared from ethyl- (2S) -2 - [[(2S, 5R) -2- (cyclopropylmethoxycarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3- en-6-yl] oxy] -2-fluoroacetate (Intermediate 115, 15 mg, 0.04 mmol) following the procedure for Example 66 to give (4.5 mg, 32.5%) after lyophilization of a white solid. MS: 344 ES + (C ₁₄ H ₁₈ FN ₃ O ₆ ).

'And NMR (300 MHz, D2O) δ: 0.00 (m, 2H); 0.28 (m, 2H); 0.81 (m, 1H); 1.38 (s, 3H); 3.02 (m, 1H); 3.35 (m, 1H); 3.42 (m, 2H); 3.82 (m, 1H); 3.92 (s, 1H); 5.37-5.55 (d, 1H); 5.95 (m, 1H).

Intermediate 116. (2S, 5R) -N- (3-Amino-3-oxopropyl) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2 .1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 200 mg, 0.64 mmol) and alaninamide hydrochloride (119.6 mg, 0.96 mmol) according to the procedure for Intermediate 101 to give (140 mg, 57.2%) a white solid. MS: 383 ES + (C ₁₇ H ₃₀ N ₄ O ₄ Si).

Intermediate 117. (2S, 5R) -N- (3-amino-3-oxopropyl) -6-hydroxy-3-methyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-ene-2- carboxamide

The title compound was prepared from (2S, 5R) -N- (3-amino-3-oxopropyl) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 116, 140 mg, 0.37 mmol) according to the procedure for Intermediate 102 to give a residue. MS: 269 ES + (C _n H ₁₆ N ₄ O ₄ ).

Intermediate 118. Ethyl- (2S) -2 - [[(2S, 5R) -2 - [(3-amino-3-oxopropyl) carbamoyl] -3methyl-7-oxo-1,6-diazabicyclo [3.2.1 ] oct-3-en-6-yl] oxy] -2-fluoroacetate

The title compound was prepared from (2S, 5R) -N- (3-amino-3-oxopropyl) -6-hydroxy-3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene- 2-carboxamide (Intermediate 117, 90 mg, 0.34 mmol), K ₂ CO ₃ (139.1 mg, 1.01 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174 , 0.12 ml, 1.01 mmol) according to the procedure for Intermediate 103 to give (26.0 mg, 20.8%) a white solid. The compound was a mixture of diastereomers in a 1: 9 ratio. MS: 373 ES + (C15H21FN4O6).

Example 71 (2S) -2 - [[(2S, 5R) -2 - [(3-Amino-3-oxopropyl) carbamoyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetic acid lithium salt

- 84 037916

The title compound was prepared from ethyl- (2S) -2 - [[(2S, 5R) -2 - [(3-amino-3-oxopropyl) carbamoyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] -2-fluoroacetate (Intermediate 118, 25 mg, 0.07 mmol) according to the procedure for Example 66 to give (8 mg, 34.6%) after lyophilization of a white solid. MS: 345 ES + (C ₁₃ H ₁₇ FN ₄ O ₆ ).

¹ H NMR (300 MHz, D2 O) δ: 1.62 (s, 3H); 2.49 (m, 2H); 3.29 (M, 1H); 3.42 (m, 1H); 3.50 (m, 2H); 4.10 (m, 1H); 4.30 (s, 1H); 5.65-5.82 (d, 1H); 6.22 (m, 1H).

Intermediate 119: 2 - [[(2S) -5-oxopyrrolidin-2-yl] methoxy] isoindoline-1,3-dione

A solution of triphenylphosphine (3.83 g, 14.6 mmol) in THF (20 ml). The suspension was stirred at -10 ° C. After 1 h, a solution of (S) - (+) - 5- (hydroxymethyl) -2-pyrrolidinone) (0.80 g, 6.95 mmol) in THF (10 ml) was added dropwise, followed by a solution of N-hydroxyphthalimide ( 1.13 g, 6.95 mmol) in THF (10 ml). The reaction mixture was allowed to warm to room temperature and was stirred for 2 days. The reaction mixture was concentrated. Chromatography on silica gel (0-100% EtOAc / hexane, then 10% MeOH / DCM) afforded the title compound (1.5 g, 83%) as a pale yellow solid. MS: 261 ES + (C ₁₃ H ₁₂ N ₂ O ₄ ).

Intermediate 120. (5S) -5- (Aminooxymethyl) pyrrolidin-2-one

To a solution of 2 - [[(2S) -5-oxopyrrolidin-2-yl] methoxy] isoindoline-1,3-dione (Intermediate 119, 1.5 g, 5.76 mmol) in DCM (70 ml) at room temperature was added hydrazine monohydrate (0.84 ml, 17.3 mmol). The reaction mixture was stirred at room temperature for 1 hour, then washed with water (3x20 ml). The aqueous layer was concentrated. Elution on a sepabead column provided the title compound (0.60 g, 79.9%) as a white solid. MS: 163 ES + (C ₈ H ₆ N ₂ O ₂ ).

Intermediate 121. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N - [(5oxopyrrolidin-2-yl) methoxy] -1,6-diazabicyclo [ 3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 250 mg, 0.80 mmol) and 5- (aminooxymethyl) pyrrolidin-2-one (Intermediate 120, 156.2 mg, 1.2 mmol) according to the procedure for Intermediate 101 to give (55 mg, 16.2%) white solid. MS: 425 ES + (C ₁₉ H ₃₂ N ₄ O ₅ Si).

Intermediate 122. (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N - [(5-oxopyrrolidin-2-yl) methoxy] -1,6-diazabicyclo [3.2.1] oct-3 -en-2-car boxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-N - [[(2S) -5-oxopyrrolidin-2-yl] methoxy] - 1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 121, 55 mg, 0.13 mmol) according to the procedure for Intermediate 102 to give a light yellow gum. MS: 311 ES + (C ₁₃ H ₁₈ N ₄ O ₅ ).

Intermediate 123. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [(5-oxopyrrolidin-2-yl) methoxycarbamoyl1-1,6- diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate

- 85 037916

O

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N - [(5oxopyrrolidin-2-yl) methoxy] -1,6-diazabicyclo [3.2.1] oct-3 -ene-2-carboxamide (Intermediate 122, 35 mg, 0.11 mmol) and ethyl (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.04 mL, 0.34 mmol) according to the procedure for Intermediate 103 to give (15 mg, 32.1%) a sticky solid. The compound was a mixture of diastereomers in a 1: 4 ratio. MS: 415 ES + (C17H23FN4O7).

Example 72. (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [(5-oxopyrrolidin-2-yl) methoxycarbamoyl] 1,6-diazabicyclo [3.2 .1] oct-3-en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [(5oxopyrrolidin-2-yl) methoxycarbamoyl] -1.6 -diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 123, 15 mg, 0.036 mmol) according to the procedure for Example 66 to obtain (7.0 mg, 40%) after lyophilization of white solid matter. MS: 387 ES + (C ₁₅ H ₁₉ FN ₄ O ₇ ).

'And NMR (300 MHz, D2O) δ: 1.64 (s, 3H); 1.82 (m, 1H); 2.25 (m, 1H); 2.38 (m, 2H); 3.24 (m, 1H); 3.76 (m, 2H); 3.96 (m, 2H); 4.09 (m, 2H); 5.63-5.82 (d, 1H); 6.17 (m, 1H).

Intermediate 124. [(2S) -5-Oxopyrrolidin-2-yl] methyl 4-methylbenzenesulfonate o / = \ / O - ^f ° n

To a stirred solution of (S) - (+) - 5- (hydroxymethyl) -2-pyrrolinone (2.0 g, 17.4 mmol) and paratoluenesulfonyl chloride (4.17 g, 21.9 mmol) in CH ₂ Cl ₂ ( 50 ml) at 0 ° C were added dimethylaminopyridine (111.4 mg, 0.91 mmol) and triethylamine (3.05 ml, 21.9 mmol). The resulting mixture was allowed to warm to room temperature. pace. and stirred for 12 hours. Then the reaction was stopped by adding water and the aqueous layer was extracted with CH ₂ Cl ₂ . The combined organic extracts were washed with 1N HCl. HCl solution and dried over anhydrous Na ₂ SO ₄ . Removal of solvent under reduced pressure and flash chromatography (2.5% MeOH in DCM) afforded the title compound (4.58 g, 93.2%) as a white solid. MS: 270 ES + (C ₁₂ H ₁₅ NO ₄ S).

Intermediate 125.2 - [(2S) -5-Oxopyrrolidin-2-yl] acetonitrile

KCN (2.76 g, 41 , 8 mmol). The solution was heated at 85 ° C for 18 hours. The solution was then diluted with acetonitrile (200 ml), filtered through celite and concentrated in vacuo. The residue was purified by flash chromatography on silica gel (9: 1 DCM / MeOH) to afford the title compound (1.8 g, 86.8%) as a white solid. MS: 125 ES + (C ₆ H ₈ N ₂ O).

Intermediate 126. tert-Butyl N- [2 - [(2S) -5-oxopyrrolidin-2-yl] ethyl] carbamate

To a stirred solution of 2 - [(2S) -5-oxopyrrolidin-2-yl] acetonitrile (Intermediate 125, 300 mg, 2.42 mmol) in methanol (15 ml) at 0 ° C was added di-tert-butyl dicarbonate (1.05 g, 4.83 mmol) and NiCl ₂ , 6 H ₂ O (57.4 mg, 0.24 mmol). Then NaBH ₄ (0.64 g, 16.9 mmol) was added over 30 min. The reaction mixture was warmed up to room temperature. pace. and stirred for 1 h. Hunig base (0.42 mL, 2.42 mmol) was added, then stirred for 30 min. The solvent was removed. The residue was dissolved in EtOAc and washed with sat. NaHCO ₃ solution, brine, dried over MgSO ₄ , filtered and concentrated to give the title compound (0.50 g, 90.6%) as a white solid. MS: 229 ES + (C11H20N2O3).

- 86 037916

Intermediate 127. (5S) -5- (2-Aminoethyl) pyrrolidin-2-one ^ nh ₂ ° H

To a solution of tert-butyl N- [2 - [(2S) -5-oxopyrrolidin-2-yl] ethyl] carbamate (Intermediate 126, 500 mg, 2.19 mmol) in DCM (2.5 mL) was added trifluoroacetic acid acid (1.15 g, 10.9 mmol). Then the reaction mixture was stirred at room temperature. pace. within 2 hours. The solvent was removed to afford the title compound as a TFA salt. MS: 129 ES + (C ₆ H ₁₂ N ₂ O).

Intermediate 128. (2S, 5R) -6-tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N- [2- (5oxopyrrolidin-2-yl) ethyl] -1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 300 mg, 0.96 mmol) and the TFA salt of 5- (2-aminoethyl) pyrrolidin-2-one (Intermediate 127, 0.35 g, 1.44 mmol) according to the procedure for Intermediate 101 to give (120 mg, 29.6%) a white solid. MS: 423 ES + (C ₂₀ H ₃₄ N ₄ O ₄ Si).

Intermediate 129. (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N- [2- (5-oxopyrrolidin-2-yl) ethyl] -1,6-diazabicyclo [3.2.1] oct -3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-N- [2- (5-oxopyrrolidin-2-yl) ethyl] -1, 6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 128, 120 mg, 0.28 mmol) according to the procedure for Intermediate 102 to give a white gum. MS: 309 ES + (C ₁₄ H ₂₀ N4O4).

Intermediate 130. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [(5-oxopyrrolidin-2-yl) methoxycarbamoyl] - 1.6 -diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N- [2- (5-oxopyrrolidin-2-yl) ethyl] -1,6-diazabicyclo [3.2.1 ] oct-3-ene-2-carboxamide (Intermediate 129, 80 mg, 0.26 mmol), K ₂ CO ₃ (179.3 mg, 1.3 mmol) and ethyl- (2S) -2-bromo- 2-fluoroacetate (Intermediate 174, 0.12 mL, 0.78 mmol) according to the procedure for Intermediate 103 to give (30 mg, 28%) a white solid. The compound was a mixture of diastereomers in a 1: 4 ratio. MS: 413 ES + (C ₁₈ H ₂₅ FN ₄ O ₆ ).

Example 73. (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- [2- (5-oxopyrrolidin-2-yl) ethylcarbamoyl] 1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- [2- (5oxopyrrolidin-2-yl) ethylcarbamoyl] -1 , 6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 130, 30 mg, 0.07 mmol) according to the procedure for example 66 to obtain after lyophilization (8.0 mg, 25.8%) of a white solid. MS: 385 ES + (C ₁₆ H ₂₁ FN ₄ O ₆ ).

¹ H NMR (300 MHz, D2 O) δ: 1.69 (s, 3H); 1.77 (m, 3H); 2.36 (m, 2H); 3.30 (m, 3H); 3.44 (m, 1H); 3.76 (m, 2H); 4.11 (m, 1H); 4.31 (m, 1H); 5.63-5.82 (d, 1H); 6.23 (m, 1H).

Intermediate 131. ((2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N- (3sulfamoylpropyl) -1,6-diazabicyclo [3.2.1] oct- 3-ene-2-carboxamide

- 87 037916

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 174, 300 mg, 0.96 mmol) and 3-aminopropane-1-sulfonamide hydrochloride (251 mg, 1.44 mmol) according to the procedure for Intermediate 101 to give (198 mg, 47.7%) a white solid matter. MS: 433 ES + (C17H32N4O5S1S).

Intermediate 132. (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N- (3-sulfamoylpropyl) -1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-N- (3-sulfamoylpropyl) -1,6-diazabicyclo [3.2.1] oct -3-ene-2-carboxamide (Intermediate 131, 200 mg, 0.46 mmol) according to the procedure for Intermediate 102 to give a white gum. MS: 319 ES + (C ₁₁ H ₁₈ N ₄ O ₅ S).

Intermediate 133. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (3-sulfamoylpropylcarbamoyl) -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N- (3-sulfamoylpropyl) -1,6-diazabicyclo [3.2.1] oct-3-ene-2- carboxamide (Intermediate 132, 100 mg, 0.31 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.11 mL, 0.94 mmol) according to the procedure for Intermediate 103 c obtaining (22 mg, 16.7%) of a white solid. The compound was a mixture of diastereomers in a 1: 4 ratio. MS: 423 ES + (C15H23FN4O7S).

Example 74. (2S) -2- (Sec-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (3-sulfamoylpropylcarbamoyl) -1,6-diazabicyclo [3.2.1] oct- 3-en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- (3sulfamoylpropylcarbamoyl) -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 133, 22 mg, 0.05 mmol) according to the procedure for Example 66 to give a white solid after lyophilization (8.0 mg, 37%). MS: 395 ES + (C ₁₃ H ₁₉ FN ₄ O ₇ S).

1H NMR (300 MHz, D ₂ O) δ: 1.69 (s, 3H); 2.04 (m, 2H); 3.35 (m, 6H); 4.11 (m, 1H); 4.34 (m, 1H); 5.655.83 (d, 1H); 6.22 (m, 1H).

Intermediate 134. tert-Butyl N- [2- (sulfamoylamino) ethyl] carbamate

A solution of tert-butyl-N- (2-aminoethyl) carbamate (2.0 g, 12.5 mmol) and sulfamide (2.0 g, 24.9 mmol) in dioxane (10 ml) was stirred at 90 ° C for 5 h. Then, the mixture was filtered to remove insoluble matter, and the filtrate was concentrated under reduced pressure. The residue was then dissolved in EtOAc, washed three times with dilute HCl solution, then brine, dried over MgSO ₄ , filtered and concentrated to give the title compound (1.2 g, 40.2%) as a yellow oil.

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.39 (s, 9H); 2.92 (m, 2H); 3.04 (m, 6H); 6.50 (m, 3H); 6.75 (m, 1H).

Intermediate 135.1-Amino-2- (sulfamoylamino) ethane TFA salt n ⁿ

Η ₂ Ν Η nh ₂

- 88 037916

To a solution of tert-butyl N- [2- (sulfamoylamino) ethyl] carbamate (Intermediate 134,

1.2 g, 5.01 mmol) in DCM (5 ml) was added trifluoroacetic acid (5.72 g, 50.1 mmol). The reaction mixture was stirred at room temperature for 2 hours. The solvent was removed to afford the title compound as a yellow TFA salt. MS: 140 ES + (C ₂ H ₉ N ₃ O ₂ S).

Intermediate 136. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N- [2 (sulfamoylamino) ethyl] -1,6-diazabicyclo [3.2.1 ] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 300 mg, 0.96 mmol) and the TFA salt of 1-amino-2- (sulfamoylamino) ethane (Intermediate 135, 365 mg, 1.44 mmol) according to the procedure for Intermediate 101 to give (117 mg, 28.1%) of a white solid. MS: 434 ES + (C ₁₆ H ₃₁ N ₅ O ₅ SiS).

Intermediate 137. (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N- [2- (sulfamoylamino) ethyl] - 1,6-diazabicyclo [3.2.1] oct-3-en-2 -carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-N- [2- (sulfamoylamino) ethyl] -1,6-diazabicyclo [3.2. 1] oct-3-ene-2-carboxamide (Intermediate 136, 117 mg, 0.27 mmol) according to the procedure for Intermediate 102 to give the title compound as a white gum. MS: 320 ES + (C ₁₀ H ₁₇ N ₅ O ₅ S)

Intermediate 138. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- [2- (sulfamoylamino) ethylcarbamoyl] - 1,6-diazabicyclo [3.2 .1] oct-3-en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N- [2 (sulfamoylamino) ethyl] -1,6-diazabicyclo [3.2.1] oct-3-ene- 2-carboxamide (Intermediate 137, 80 mg, 0.25 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.089 mL, 0.75 mmol) according to the procedure for Intermediate 103 c obtaining (22.0 mg, 16.5%) as a white solid. The compound was a mixture of diastereomers in a 3: 7 ratio. MS: 424 ES + (C ₁₄ H ₂₂ FN ₅ O ₇ ).

Example 75 (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- [2- (sulfamoylamino) ethylcarbamoyl] -1.6 diazabicyclo [3.2.1] oct- 3-en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2- [2 (sulfamoylamino) ethylcarbamoyl] - 1,6-diazabicyclo [ 3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 138, 30 mg, 0.071 mmol) according to the procedure for Example 66 to give (13.0 mg, 44.1%) a white solid. MS: 396 ES + (C ₁₂ H ₁₈ FN ₅ O ₇ S).

¹ H NMR (300 MHz, D2 O) δ: 1.71 (s, 3H); 3.21-3.47 (m, 6H); 4.11 (m, 1H); 4.35 (m, 1H); 5.64-5.84 (d, 1H); 6.22 (m, 1H).

Intermediate 139. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-en-2-carbonitrile

To a solution of (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-ene-2-carboxamide (Intermediate 192 , 3.0 g, 9.63 mmol) in DCM (50 ml) at room temperature was added Burgess reagent (3.44 g, 14.4 mmol) in portions over 2 h. The reaction mixture was stirred for another 16 h, then washed twice with a mixture of brine and water in a ratio of 1: 1. The organics were dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-25% ethyl acetate / hexanes) afforded the title compound (2.3 g, 81.3%) as a white solid. MS: 294 ES + (C ₁₄ H ₂₃ N ₃ O ₂ Si).

Intermediate 140. tert-Butyl-N - [[(2S) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-yl] methyl] carbamate

To a stirred solution of (2S) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-carbonitrile (Intermediate 139, 1.2 g, 4.09 mmol) in methanol (100 ml) at 0 ° С, di-tert-butyl dicarbonate (1.78 g, 8.18 mmol) and NiCl ₂ , 6 H ₂ O ( 97.2 mg, 0.41 mmol). Then NaBH ₄ (1.08 g, 28.6 mmol) was added over 30 min. Then the reaction mixture was then warmed up to room temperature. pace. and stirred for 1 hour. Hunig's base (0.71 ml, 4.09 mmol) was added, then stirred for 30 minutes. The solvent was removed. The residue was dissolved in EtOAc and washed with sat. NaHCO ₃ solution, brine, dried over MgSO ₄ , filtered and concentrated. The residue was purified by silica gel chromatography (40 g, 0% -35% EtOAc / hexane) to give the title compound (0.58 g, 35.7%) as a white solid. MS: 398 ES + (C ₁₉ H ₃₅ N ₃ O ₄ Si).

Intermediate 141. tert-Butyl-N - [[(2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-yl] methyl] car bamat

The title compound was prepared from tert-butyl N - [[(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1 ] oct-3-en-2-yl] methyl] carbamate (Intermediate 140, 80 mg, 0.20 mmol) according to the procedure for Intermediate 102 to give a white gum. MS: 284 ES + (C ₁₃ H ₂₁ N ₃ O ₄ ).

Intermediate 142. Ethyl- (2S) -2 - (((2S, 5R) -2 - (((tert-butoxycarbonyl) amino) methyl) -3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetate

The title compound was prepared from tert-butyl N - [[(2S, 5R) -6-hydroxy-3-methyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-en-2-yl] methyl] carbamate (Intermediate 141, 50 mg, 0.18 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.063 mL, 0.53 mmol) according to the procedure for Intermediate 103 c obtaining (50 mg, 73.1%) of a white solid. The compound was a mixture of diastereomers in a 1: 9 ratio. MS: 388 ES + (C17H26FN3O6).

Example 76. (2S) -2 - [[(2S, 5R) -2 - [(tert-Butoxycarbonylamino) methyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3- en-6-yl] oxy] -2-fluoroacetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2 - (((2S, 5R) -2 - (((tert-butoxycarbonyl) amino) methyl) -3-methyl-7-oxo-1,6-diazabicyclo [ 3.2.1] oct-3-en-6-yl) oxy) -2-fluoroacetate (Intermediate 142, 50 mg, 0.13 mmol) according to the procedure for example 66 to obtain after lyophilization (35 mg, 67.9% ) a white solid. MS: 360 ES + (C ₁₅ H ₂₂ FN ₃ O ₆ ).

'H NMR (300 MHz, D ₂ O) δ: 1.43 (s, 9H); 1.65 (s, 3H); 3.17-3.36 (m, 3H); 3.49 (m, 1H); 3.74 (m, 1H); 4.05 (m, 1H); 5.62-5.81 (d, 1H); 6.08 (m, 1H).

Example 77. (2S) -2 - [[(2S, 5R) -2- (Aminomethyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl ] hydroxy] -2-fluoroacetic acid

- 90 037916

To a solution of (2S) -2 - [[(2S, 5R) -2 - [(tert-butoxycarbonylamino) methyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -6-yl] oxy] -2-fluoroacetic acid (Example 76, 25 mg, 0.07 mmol) in DCM (1 ml) at 0 ° C. was added trifluoroacetic acid (0.79 g, 6.96 mmol). The reaction mixture was stirred for 2 hours, then concentrated. The residue was dissolved in pH 7 buffer, then loaded onto a sepabead column (first saturated with water, then ACN, then washed with water) and eluted (0-2.5% ACN / water) to give, after lyophilization, the title compound (8 mg , 37.7%) as a white solid. MS: 260 ES + (C ₁₀ H ₁₄ FN ₃ O4).

'And NMR (300 MHz, D2O) δ: 1.64 (s, 3H); 3.19-3.30 (m, 3H); 3.44 (m, 1H); 3.96 (m, 1H); 4.09 (m, 1H); 5.64-5.84 (d, 1H); 6.16 (m, 1H).

Intermediate 143. (2S, 5R) -2- (Aminomethyl) -6- [tert-butyl (dimethyl) silyl] oxy-3methyl-1,6-diazabicyclo [3.2.1] oct-3-en-7-one

To a solution of tert-butyl-N - [[(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-2-yl] methyl] carbamate (Intermediate 140, 100 mg, 0.25 mmol) in DCM (5 ml), ZnBr ₂ (170 mg, 0.75 mmol) was added at 0 ° C. The reaction mixture was stirred at room temperature. pace. within 16 hours, then concentrated and used in the next step without purification. MS: 298 ES + (C14H27N3O2Si).

Intermediate 144. N - [[(2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-ene-2 -yl] methyl] acetamide

To a solution of (2S, 5R) -2- (aminomethyl) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-1,6-diazabicyclo [3.2.1] oct-3-en-7-one (Intermediate 143, 74 mg, 0.25 mmol) in pyridine (1 ml) at 0 ° C was added Ac ₂ O (168 mg, 0.75 mmol). The reaction mixture was stirred at room temperature. pace. for 30 min, then washed with water, sat. NaHCO ₃ solution, brine, dried over MgSO ₄ , filtered and concentrated. The residue was purified by silica gel chromatography (0-100% EtOAc / hexane) to afford the title compound (70 mg, 78.7%) as a white solid. MS: 340 ES + (C16H29N3O3Si).

Intermediate 145. N - [[(2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-yl] methyl] acetamide

Lo I

X λ --- ^N 4

OH OH

The title compound was prepared from N - [[(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-yl] methyl] acetamide (Intermediate 144, 70 mg, 0.21 mmol) according to the procedure for Intermediate 102 to give a white gum. MS: 226 ES + (C ₁₀ H ₁₅ N ₃ O ₃ ).

Intermediate 146. Ethyl- (2S) -2 - [[(2S, 5R) -2- (acetamidomethyl) -3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6- yl] oxy] -2-fluoroacetate ⁰ ° o ⁷

The title compound was prepared from N - [[(2S, 5R) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-yl] methyl] acetamide (Intermediate 145, 40 mg, 0.18 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.063 ml, 0.53 mmol) according to the procedure for Intermediate 103 to be obtained after lyophilization (15 mg, 25.6%) of a white solid. The compound was a mixture of diastereomers in the ratio 15:85. MS: 330 ES + (C ₁₄ H ₂₀ FN ₃ O ₅ ).

Example 78. (2S) -2 - [[(2S, 5R) -2- (Acetamidomethyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl] oxy ] -2-fluoroacetic acid lithium salt

- 91 037916

The title compound was prepared from ethyl- (2S) -2 - [[(2S, 5R) -2- (acetamidomethyl) -3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en- 6-yl] oxy] -2-fluoroacetate (Intermediate 146, 15 mg, 0.05 mmol) according to the procedure for Example 66 to give a white solid after lyophilization (10 mg, 65.6%). MS: 302 ES + (C ₁₂ H ₁₆ FN ₃ O ₅ ).

¹ H NMR (300 MHz, D ₂ O) δ: 1.64 (s, 3H); 1.99 (s, 3H); 3.19 (m, 1H); 3.34 (m, 2H); 3.64 (m, 1H); 3.79 (m, 1H); 4.05 (m, 1H); 5.63-5.81 (d, 1H); 6.09 (m, 1H).

Intermediate 147. Ethyl- (2S) -2 - [[(2S, 5R) -2- (aminomethyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene- 6-yl] oxy] -2-fluoroacetate TFA salt

To a solution of ethyl- (2S) -2 - [[(2S, 5R) -2 - [(tert-butoxycarbonylamino) methyl] -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] -2-fluoroacetate (Intermediate 142, 97.4 mg, 0.25 mmol) in DCM (2 ml) at 0 ° C was added trifluoroacetic acid (2.87 g, 25.2 mmol). The reaction mixture was stirred at 0 ° C for 2 h, then concentrated to give the title compound as a TFA salt. MS: 288 ES + (C ₁₂ H ₁₈ FN ₃ O ₄ ).

Intermediate 148.tert-Butyl-N-chlorosulfonylcarbamate

To a stirred solution of tert-butanol (1.9 ml, 20 mmol) in CH ₂ Cl ₂ (12 ml) at 0 ° C. was added chlorosulfonyl isocyanate (1.4 ml, 15 mmol) dropwise over 10 min. After stirring at 0 ° C for 5 min, the reaction mixture was warmed up to room temperature. pace. and stirred for 20 minutes. The reaction mixture was concentrated in vacuo to one third of the original volume. The flask was placed back in a bath at 0 ° C and the product crystallized out of solution. After 50 min, the product was collected by filtration and washed with hexanes to give the title compound (2.8 g, 80.7%) as a white solid.

Intermediate 149. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [[(2,2,2-trifluoroacetyl) amino] methyl] - 1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate

To a solution of TFA salt ethyl- (2S) -2 - [[(2S, 5R) -2- (aminomethyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -6-yl] oxy] -2-fluoroacetate (Intermediate 147, 72 mg, 0.25 mmol) in DCM (2 ml) at 0 ° C was added tert-butyl N-chlorosulfonylcarbamate (Intermediate 148, 54.0 mg, 0.25 mmol) and triethylamine (63.4 mg, 0.63 mmol). The reaction mixture was stirred at room temperature. pace. for 30 min, then diluted with DCM, washed with water, brine, dried over MgSO ₄ , filtered and concentrated. The residue was purified by flash chromatography (0-100% EtOAc / hexane) to afford the title compound (38 mg, 39.6%) as a white solid. MS: 384 ES + (C14H17F4N ₃ O5).

Example 79. (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [[(2,2,2-trifluoroacetyl) amino] methyl] -1.6 diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 [[(2,2,2-trifluoroacetyl) amino] methyl] -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 149, 38 mg, 0.10 mmol) according to the procedure for example 66 to obtain after lyophilization (25 mg, 60.3%) of a white solid. MS: 356 ES + (C ₁₂ H ₁₃ FN ₃ O ₅ ).

¹ H NMR (300 MHz, D ₂ O) δ: 1.66 (s, 3H); 3.22 (m, 1H); 3.36 (m, 1H); 3.59 (m, 1H); 3.73 (m, 1H); 3.86 (m, 1H); 4.08 (m, 1H); 5.62-5.83 (d, 1H); 6.11 (m, 1H).

Intermediate 150. (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-N- (cyanomethyl) -3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carboxamide

- 92 037916

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 700 mg,

2.24 mmol) and aminoacetonitrile hydrochloride (207 mg, 2.24 mmol) according to the procedure for Intermediate 101 to give (302 mg, 38.4%) a white solid. MS: 351 ES + (C16H26N4O3Si).

Intermediate 151. (2S, 5R) -N- (Cyanomethyl) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-N (cyanomethyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct- 3-ene-2-carboxamide (Intermediate 150, 300 mg, 0.86 mmol) according to the procedure for Intermediate 102 to give a residue. MS: 237 ES + (C10H12N4O3).

Example 80. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (cyanomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetate

To a solution of (2S, 5R) -N- (cyanomethyl) -6-hydroxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-2carboxamide (Intermediate 151, 239 mg , 1.01 mmol) in THF (3 ml) and DMF (0.3 ml) at 40 ° С, DBU (0.18 ml, 1.22 mmol) and ethyl- (2S) -2-bromo-2- fluoroacetate (Intermediate 174, 0.18 ml, 1.52 mmol). The reaction mixture was stirred at -40 ° C for 30 min, then diluted with ethyl acetate and washed three times with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. The residue was purified by flash chromatography (0-100% EtOAc / hexane) to give a 1: 4 mixture of diastereomers. The diastereomers were separated by reverse phase HPLC (T3 column, 20% -50% ACN / water, 10 min) to give Example 80 (64 mg, 18.2%) and Example 81 (8 mg, 2.3%) in the form of white solids. MS: 341 ES + (C ₁₄ H ₁₇ FN ₄ O ₅ ).

1H NMR (300 MHz, DMSO-de) δ: 1.27 (m, 3H); 1.64 (s, 3H); 3.13 (m, 1H); 3.61 (m, 1H); 3.98 (m, 1H); 4.21 (m, 5H); 6.06-6.26 (m, 1H); 6.10 (m, 1H); 9.18 (m, 1H).

Example 81. Ethyl- (2R) -2-fluoro-2 - [[(2S, 5R) -2- (cyanomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetate

MS: 341 ES + (C14H17FN4O5).

1H NMR (300 MHz, DMSO-de) δ: 1.21 (m, 3H); 1.65 (s, 3H); 3.12 (m, 1H); 3.60 (m, 1H); 4.07 (m, 1H); 4.22 (m, 5H); 6.08 (m, 1H); 6.15-6.33 (m, 1H); 9.16 (m, 1H).

Example 82 (2S) -2-Fluoro-2 - [[(2S, 5R) -2- (cyanomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (cyanomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Example 80, 50 mg, 0.15 mmol) according to the procedure for Example 66 to give a white solid after lyophilization (32 mg, 66.3%). MS 313 ES + (C ₁₂ H ₁₃ FN ₄ O ₅ ).

1 H NMR (300 MHz, D ₂ O) δ: 1.74 (s, 3H); 3.35 (m, 2H); 4.12 (m, 1H); 4.25 (m, 2H); 4.42 (m, 1H); 5.635.83 (d, 1H); 6.23 (m, 1H).

Example 83. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (cyanomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetate

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Intermediate 138 (230 mg) was separated using a reverse phase preparative

HPLC (T3 column, ACN / water 20-50% for 10 min) to give Example 83 (104 mg) and Example (7 mg) as white solids. MS: 424 ES + (C ₁₄ H ₂₂ FN ₅ O ₇ S).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 1.27 (m, 3H); 1.63 (s, 3H); 2.97 (m, 2H); 3.10 (m, 1H); 3.26 (m, 2H); 3.73 (m, 1H); 3.96 (m, 1H); 4.21 (m, 3H); 6.06-6.26 (m, 2H); 6.56 (m, 3H); 8.45 (m, 1H).

Example 84. Ethyl- (2R) -2-fluoro-2 - [[(2S, 5R) -2- (cyanomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetate

MS: 424 ES + (C ₁₄ H ₂₂ FN ₅ O ₇ S).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.21 (m, 3H); 1.62 (s, 3H); 2.95 (m, 2H); 3.07 (m, 1H); 3.24 (m, 2H); 3.71 (m, 1H); 4.06 (m, 1H); 4.21 (m, 3H); 6.03 (m, 1H); 6.14-6.31 (m, 1H); 6.55 (m, 3H); 8.42 (m, 1H).

Intermediate 152. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-N- (hydroxymethyl) -3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carboxamide

To a solution of (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-ene-2-carboxamide (Intermediate 192 , 1.0 g, 3.21 mmol) was added paraformaldehyde (1.45 g, 16.1 mmol) in 1,4-dioxane (10 ml). The reaction mixture was heated to 90 ° C for 16 h in a microwave oven. The solvent was removed. Chromatography on silica gel (0-80% EtOAc / Hexane) afforded the title compound (0.62 g, 56.5%) as a white solid. MS: 342 ES + (C ₁₅ H ₂₇ N ₃ O ₄ Si).

Intermediate 153. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (hydroxymethylcarbamoyl) -3methyl-7-oxo -1,6-diazabicyclo [3.2.1] oct-3 - en-6-yl] oxy] acetate

To a solution of (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-N- (hydroxymethyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carboxamide (Intermediate 152, 230 mg, 0.67 mmol) and cerium (III) chloride (166 mg, 0.67 mmol) in THF (3 ml) at -78 ° C was added TBAF (0.67 ml , 0.67 mmol) (1M in THF). The mixture was stirred for about 10 minutes. Ethyl (2S) -2-bromo-2fluoroacetate (Intermediate 174, 24.4 mg, 0.67 mmol) was added to the reaction mixture. After 5 min, the reaction mixture was partitioned between water and EtOAc. The organic layer was collected, washed with water, brine, dried over MgSO ₄ , filtered and concentrated. Chromatography on silica gel (0-80% EtOAc / hexane) gave a 1: 3 mixture of two diastereomers, 180 mg.

Example 85. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (hydroxymethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetate

Intermediate 153 (180 mg) was separated by reverse phase preparative HPLC (T3 column, ACN / water 20-50% for 10 min) to give Example 85 (43 mg) and Example 86 (2.6 mg) as white solids. MS: 332 ES + (C ₁₃ H ₁₈ FN ₃ O ₆ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.28 (m, 3H); 1.61 (s, 3H); 3.09 (m, 1H); 3.79 (m, 1H); 3.96 (m, 1H); 4.25 (m, 3H); 4.54 (m, 2H); 5.68 (m, 1H); 6.06-6.24 (m, 2H); 8.99 (m, 1H).

Example 86. Ethyl- (2R) -2-fluoro-2 - [[(2S, 5R) -2- (hydroxymethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetate

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MS: 332 ES + (C13H18FN3O6).

1H NMR (300 MHz, DMSO-d6) δ: 1.24 (m, 3H); 1.61 (s, 3H); 2.97 (m, 1H); 3.35 (m, 1H); 4.06 (m, 1H);

4.26 (m, 3H); 4.59 (m, 2H); 5.52 (m, 1H); 6.03 (m, 1H); 6.06-6.24 (m, 1H); 8.99 (m, 1H).

Example 87. (2S) -2-Fluoro-2 - [[(2S, 5R) -2- (hydroxymethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo

[3.2.1] oct-3-en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (hydroxymethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Example 85, 15 mg, 0.05 mmol) according to the procedure for Example 66 to give, after lyophilization (8 mg, 55.3%), a white solid.

MS: 304 ES + (C11H14FN3O6).

1H NMR (300 MHz, D2O) δ: 1.77 (s, 3H); 3.42 (m, 2H); 4.17 (m, 1H); 4.41 (m, 1H); 4.79 (m, 2H); 5.705.89 (d, 1H); 6.28 (m, 1H).

Intermediate 154.tert-Butyl-N- (isocyanatomethyl) carbamate

To a stirred solution of BOC-GLY-OH (10 g, 57.08 mmol) in THF (200 ml) at 0 ° C, methyl chloroformate (5.29 ml, 68.5 mmol) was added dropwise followed by triethylamine ( 9.55 ml, 68.5 mmol). A white precipitate formed immediately. The mixture was stirred for 45 min before adding NaN ₃ (5.58 g, 85.6 mmol) in water (10 ml) at 0 ° C. The resulting mixture was stirred at 0 ° C for 1 h, then diluted with water. The acyl azide was extracted four times with toluene (4x25 ml) and the combined organic extracts were sequentially washed with saturated sodium bicarbonate solution (2x30 ml) and water (50 ml). The organics were dried over MgSO ₄ at 0 ° C, filtered and then slowly heated with stirring until nitrogen gas evolution was observed, which continued at 59 ° C for 20 min. The temperature was raised and maintained at 64 ° C for 1.5 hours, then slowly raised to 70 ° C over 20 minutes. The solution was concentrated under reduced pressure to obtain the title compound (8.6 g, 87.5%) as a colorless oil.

Intermediate 155.tert-Butyl-N- (benzyloxycarbonylaminomethyl) carbamate

To a solution of tert-butyl N- (isocyanatomethyl) carbamate (Intermediate 154, 8.3 g, 48.2 mmol) in DCE (5 ml) at 0 ° C was added benzyl alcohol (7.48 ml, 72.3 mmol ) and triethylamine (0.67 ml, 4.82 mmol), dropwise. The reaction mixture was warmed up to room temperature. pace. and stirred for 30 minutes. The resulting white solid was collected by filtration and washed with hexane to give the title compound (5.2 g, 38.5%) as a white solid. MS: 303 ES + Na (C14H20N2O4).

Intermediate 156.tert-Butyl-N- (aminomethyl) carbamate

A solution of tert-butyl N- (benzyloxycarbonylaminomethyl) carbamate (Intermediate 155, 1.5 g, 5.35 mmol) in methanol (20 mL) was bubbled with nitrogen gas. Pd / C (10%) (150 mg) was added. The reaction mixture was degassed and then placed under a hydrogen atmosphere from a balloon for 30 minutes. The reaction mixture was filtered through celite. The filtrate was concentrated to give the title compound as a colorless oil. MS: 147 ES + (C ₆ H ₁₄ N ₂ O ₂ ).

Intermediate 157. tert-Butyl-N - [[(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3 -ene-2-carbonyl] amino] methyl] carbamate

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 350 mg, 1.12 mmol) and tert-butyl-N- (aminomethyl) carbamate (Intermediate 156, 246 mg, 1.68 mmol) according to the procedure for Intermediate 101 to give (170 mg, 34.4%) white

- 95 037916 solid. MS: 441 ES + (C ₂₀ H ₃₆ N ₄ O5Si).

Intermediate 158. (2S, 5R) -N- (aminomethyl) -6- [tert-butyl (dimethyl) sulyl] oxy-3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carboxamide

To a solution of tert-butyl-N - [[[(2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct- 3-en-2-carbonyl] amino] methyl] carbamate (Intermediate 157, 170 mg, 0.39 mmol) in DCM (3 ml) at 0 ° C was added ZnBr ₂ (261 mg, 1.16 mmol). The reaction mixture was stirred at room temperature. pace. over 16 h, then concentrated to give the title compound. MS: 341 ES + (C1 ₅ H2 ₈ N4O3Si).

Intermediate 159: (2S, 5R) -N- (acetamidomethyl) -6- [tert-butyl (dimethyl) silyl] oxy-3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carboxamide

To a solution of (2S, 5R) -N- (aminomethyl) -6- [tert-butyl (dimethyl) sulyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -2-carboxamide (Intermediate 158, 131 mg, 0.39 mmol) in pyridine (3 ml) at 0 ° C was added acetic anhydride (394 mg, 3.86 mmol). The reaction mixture was stirred at room temperature. pace. Within 30 min, then partitioned between EtOAc and water. The organic layer was washed with water, sat. NaHCO ₃ solution, brine, dried over MgSO ₄ , filtered and concentrated. Chromatography on silica gel (0-80% EtOAc / Hexane) afforded the title compound (75 mg, 50.8%) as a white solid. MS: 383 ES + (C ₁₇ H ₃₀ N ₄ O ₄ Si).

Intermediate 160. (2S, 5R) -N- (Acetamidomethyl) -6-hydroxy-3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -N- (acetamidomethyl) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct -3-ene-2-carboxamide (Intermediate 159, 75 mg, 0.20 mmol) according to the procedure for Intermediate 102 to give a white gum. MS: 269 ES + (C11H16N4O4).

Intermediate 161. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (acetamidomethylcarbamoyl) -3methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3- en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -N- (acetamidomethyl) -6-hydroxy-3-methyl-7oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 160, 50 mg, 0.19 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.067 mL, 0.56 mmol) according to the procedure for Intermediate 103 to give a white solid after elution (0-15% ACN / water) on a sepabead column (saturated with water first, then ACN, then eluted with water) and lyophilized, 21 mg, 27%. The compound was a mixture of diastereomers in a 1: 4 ratio. MS: 373 ES + (C ₁₅ H ₂₁ FN ₄ O ₆ ).

Example 88 (2S) -2-Fluoro-2 - [[(2S, 5R) -2- (acetamidomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene -6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -2- (acetamidomethylcarbamoyl) -3-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 161, 21 mg, 0.056 mmol) in water (1 ml) according to the procedure for example 66 to obtain after lyophilization (6.0 mg, 29.3%) white solid. MS: 345 ES + (C ₁₃ H ₁₇ FN ₄ O ₆ ).

¹ H NMR (300 MHz, D2 O) δ: 1.75 (s, 3H); 2.02 (s, 3H); 3.42 (m, 2H); 4.15 (m, 1H); 4.37 (m, 1H); 4.64 (m,

- 96 037916

2H); 5.70-5.89 (d, 1H); 6.27 (m, 1H).

Intermediate carbamate

162. tert-Butyl-N - [(tert-butoxycarbonylamino) methylsulfamoyl]

To a solution of tert-butyl N- (aminomethyl) carbamate (Intermediate 156, 700 mg, 4.79 mmol) in DCM (20 mL) at 0 ° C was added triethylamine (0.67 mL, 4.79 mmol). Then, tert-butyl-N-chlorosulfonylcarbamate (Intermediate 148, 1.03 g, 4.79 mmol) in DCM (5 ml) was added dropwise. The reaction mixture was stirred at 0 ° C for 30 min, then diluted with DCM, washed with water, brine, dried over MgSO ₄ , filtered and concentrated. The residue was purified by washing with DCM / hexane to give the title compound (650 mg, 41.7%) as a white solid. MS: 324 ES- (C11H ₂₃ N ₃ O ₆ S).

Intermediate 163. Amino (sulfamoylamino) methane TFA salt

H ₂ NN NH ₂ Η

To a solution of tert-butyl-N - [(tert-butoxycarbonylamino) methylsulfamoyl] carbamate (Intermediate 162, 650 mg, 2 mmol) in DCM (5 mL) at 0 ° C was added trifluoroacetic acid (2.28 g, 19.9 mmol). The reaction mixture was stirred at room temperature. pace. over 30 min then concentrated to give the title compound as a light yellow gum.

Intermediate 164. (2S, 5R) -6- [tert-Butyl (dimethyl) silyl] oxy-3-methyl-7-oxo-N [(sulfamoylamino) methyl-1,6-diazabicyclo [3.2.1] oct- 3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylic acid (Intermediate 77, 350 mg, 1.12 mmol) according to the procedure for Intermediate 101 to give (125 mg, 26.6%) a white solid. MS: 420 ES + (C ₁₅ H ₂₉ N ₅ O ₅ SiS).

Intermediate 165. (2S, 5R) -6-Hydroxy-3-methyl-7-oxo-N - [(sulfamoylamino) methyl] 1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

The title compound was prepared from (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl7-oxo-N - [(sulfamoylamino) methyl] -1,6-diazabicyclo [3.2.1] oct-3-en-2-carboxamide (Intermediate 164, 125 mg, 0.30 mmol) according to the procedure for Intermediate 102 to give a light yellow gum. MS: 306 ES + (C ₉ H ₁₅ N ₅ O ₅ S).

Intermediate 166. Ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [(sulfamoylamino) methylcarbamoyl] -1,6-diazabicyclo [3.2.1 ] oct-3-en-6-yl] oxy] acetate

The title compound was prepared from (2S, 5R) -6-hydroxy-3-methyl-7-oxo-N - [(sulfamoylamino) methyl] - 1,6-diazabicyclo [3.2.1] oct-3-en-2 -carboxamide.

(Intermediate 165, 88 mg, 0.29 mmol) and ethyl- (2S) -2-bromo-2-fluoroacetate (Intermediate 174, 0.10 ml, 0.86 mmol) according to the procedure for Intermediate 103 to give (18 mg, 12.2%) as a white solid. The compound was a mixture of diastereomers in a ratio of 17:83. MS: 410 ES + (C ₁₃ H ₂₀ FN ₅ O ₇ S).

Example 89 (2S) -2-Fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2 - [(sulfamoylamino) methylcarbamoyl] -1.6 diazabicyclo [3.2.1] oct-3 -en-6-yl] oxy] acetic acid lithium salt

The title compound was prepared from ethyl- (2S) -2-fluoro-2 - [[(2S, 5R) -3-methyl-7-oxo-2-97 037916

[(sulfamoylamino) methylcarbamoyl] -1,6-diazabicyclo [3.2.1] oct-3-en-6-yl] oxy] acetate (Intermediate 166, 18 mg, 0.040 mmol) according to the procedure for Example 66 to be obtained after lyophilization (5.0 mg, 25.3%) as a white solid. MS: 382 ES + (CnH ₁₆ F5 ₄ O ₇ S).

¹ H NMR (300 MHz, D2 O) δ: 1.76 (s, 3H); 3.40 (m, 2H); 4.16 (m, 1H); 4.41 (m, 1H); 4.63 (m, 2H); 5.705.89 (d, 1H); 6.28 (m, 1H).

Intermediate 167. Racemic 2-bromo-2-fluoroacetic acid

A solution of ethyl 2-bromo-2-fluoroacetate (3.5 kg) in tetrahydrofuran (7 liters, 2 vol.) And a solution of sodium hydroxide (830 g) in water (7 L, 2 vol.) for 1 hour. The resulting solution was stirred at 0-5 ° C for 1 hour. HCl (160 ml) was added dropwise at 0-5 ° C. Water and tetrahydrofuran were removed by concentration in vacuo. The residue was suspended in tetrahydrofuran (35 L, 10 vol.) And conc. HCl (1.57 L, 1.0 eq.). Anhydrous sodium sulfate was added and the resulting mixture was stirred for 2 hours. The solid was filtered off and washed with THF (1 Lx2). The filtrate was concentrated in vacuo to give 2-bromo-2-fluoroacetic acid (2.2 kg) as a yellow oil, which was combined with a previous batch prepared in the same way (940 g, purity: 72%) and subjected to vacuum distillation. (65-70 ° C 100 Pa) to obtain 2-bromo-2-fluoroacetic acid (2.55 kg, 67% overall yield) as a colorless oil.

1H NMR (400 MHz, CDCl ₃ ): δ 11.15 (s. 1H), 6.66 (d, 1H, J = 68 Hz).

Intermediate 168. (S) -1-Phenylethane-1-amine- (R) -2-bromo-2-fluoroacetate

A solution of 2-bromo-2-fluoroacetic acid (Intermediate 167, 2.0 kg) in 1 L of chloroform (1 vol.) Was loaded into a 10 L reactor at 0-5 ° C, to which solution (S) was added dropwise -1 phenylethanamine (1.39 kg) in 1 liter of chloroform (1 vol.). The mixture was stirred at room temperature overnight and the resulting white solid was collected by filtration to give (S) -1phenylethanamine 2-bromo-2-fluoroacetate salt (2.5 kg; ee: 6%), which was charged to a 10 L reactor, and then chloroform (5 L, 2 vol.) was added. The resulting mixture was stirred for 2 h at 50 ° C (the solid was partially dissolved in chloroform), cooled to 0 ° C and left for 2 h. The solid was collected by filtration and washed with chilled chloroform (500 ml, 0.2 vol). The recrystallization procedure was repeated 4 times to give 1.09 kg (97% ee) of the title compound as a white solid in 31% overall yield (2 steps).

Intermediate 169. Pentan-3-yl- (R) -2-bromo-2-fluoroacetate

(S) -1-phenylethanamine (R) -2-6pom-2fluoroacetate (Intermediate 168, 450 g), dichloromethane (900 ml, 2 vol.) And iPrOH (2.0 eq. .). Chlorotrimethylsilane (1.12 L) was added slowly and a white precipitate formed. The resulting mixture was stirred at room temperature overnight. The white precipitate was filtered off and the filter cake was washed with hexane (450 ml, 1 vol.). The combined filtrate was washed with water (3x100 ml). The organic solution was dried with anhydrous sodium sulfate, filtered, concentrated in vacuo. The residue was distilled (54-60 ° C, 100 Pa) to give the title compound as a colorless oil (290 g, 79% yield, 95% purity).

Ή NMR (CDCl ₃ , 400 MHz): δ 6.53 (d J = 51.2 Hz, 1H), 5.17 (m, 1H), 1.32 (m, 6H). Intermediate 170: Ethyl- (R) -2-bromo-2-fluoroacetate

In a three-necked round-bottom flask with a volume of 50 ml, purged and maintained in an inert atmosphere of nitrogen, was placed (1R) -1-phenylethan-1-amine; (2S) -2-bromo-2-fluoroacetic acid (Intermediate 168, 30 g, 107.9 mmol) and ethanol (34.7 g, 755.3 mmol). Chlorotrimethylsilane (82 g, 755.3 mmol) was then added dropwise with stirring at room temperature. The resulting solution was stirred for 4 hours at room temperature, then the reaction was stopped by adding 10 ml of water / ice. The resulting solution was extracted with 3x20 ml of petroleum ether (30-60 degrees) and the organic layers were combined. The resulting mixture was washed with 2x20 ml brine. The mixture was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was eluted with petroleum ether (30-60 degrees) on a silica gel column. This gave 10 g (50%) of the title compound as a colorless oil.

- 98 037916

1H NMR (300 MHz CDCI3): δ 6.58 (d, 1H, J = 51 Hz), 4.38 (q, 2H, J = 6 Hz), 1.38 (t, 3H, J = 6 Hz) ...

Intermediate 171. (R) -Benzyl-2-bromo-2-fluoroacetate

Chlorotrimethylsilane (60 ml, 719.12 mmol) was added portionwise to (S) -1-phenylethanamine (R) -2-6pom2-fluoroacetate (Intermediate 168, 20 g, 71.91 mmol) and phenylmethanol (60 ml, 71 , 91 mmol) at 25 ° С for 3 min under nitrogen atmosphere. The resulting solution was stirred at 25 ° C for 4 hours. The reaction mixture was diluted with heptane (500 ml), then washed with water and brine. The organics were dried over sodium sulfate, filtered and concentrated. Chromatography on silica gel (0% -10% ethyl acetate / petroleum ether) gave the title compound as a yellow oil, 17.5 g, 98%. 1H NMR (300 MHz, CDCl ₃ -d, 30 ° C) δ: 5.30 (s, 2H); 6.60 (d, 1H); 7.40 (m, 5H).

Intermediate 172. (1R) -1-Phenylethan-1-amine; (2S) -2-bromo-2-fluoroacetic acid

HE

A solution of (1R) -1-phenylethan-1-amine (107.7 g, 0.89 mol) in methanol (325 ml) was placed in a 100 ml round bottom flask. A solution of 2-bromo-2-fluoroacetic acid (Intermediate 167, 140 g, 0.89 mol) in methanol (420 ml) was then added dropwise with stirring at 0 ° C. for 30 minutes. The resulting solution was stirred overnight at room temperature, then concentrated in vacuo. The residue was diluted with CHCl ₃ (3 vol.). The solids were collected by filtration. The solid was dried in vacuo, then suspended in CHCl ₃ and heated to 60 ° C for 2 hours. The mixture was then cooled to 0 ° C and the solid was filtered off. This process was repeated 6 times. This gave 80 g (32%) of the title compound as a white solid.

1H NMR (300 MHz, d6-DMSO): δ 8.56 (brs, 3H), 7.49-7.46 (m, 2H), 7.42-7.38 (m, 2H), 7.36 -7.34 (m, 1H), 6.48 (d, 1H, J = 56 Hz), 4.39-4.34 (m, 1H), 1.48 (d, 3H, J = 6.8 Hz).

Intermediate 173. (S) -Isopropyl-2-bromo-2-fluoroacetate

(R) -1-phenylethanamine (S) -2-bromo-2-fluoroacetate (Intermediate 172, 32.0 g, 116 mmol) and isopropanol (13.9 g, 232 mmol) in DCM (64 ml). Chlorotrimethylsilane (56.4 g, 519 mmol) was then added dropwise with stirring at room temperature. The resulting solution was stirred overnight at room temperature, then the reaction was stopped by adding 100 ml of water / ice. The resulting solution was extracted with 3x100 ml of petroleum ether (30-60 degrees) and the organic layers were combined. The resulting mixture was washed with 3x70 ml brine. The mixture was dried over anhydrous sodium sulfate and concentrated. The residue was eluted with petroleum ether (30-60 degrees) on a silica gel column. This gave 19 g (83%) of the title compound as a colorless oil.

1H NMR (CDCl3, 400 MHz): δ 6.53 (d, J = 50.8 Hz, 1H), 5.17 (m, 1H), 1.32 (m, 6H).

Intermediate 174. Ethyl- (2S) -2-bromo-2-fluoroacetate

In a 50 ml three-necked round-bottom flask, purged and maintained under an inert nitrogen atmosphere, was placed (1R) -1-phenylethane-1-amine (2S) -2-bromo-2-fluoroacetic acid (Intermediate 172, 20 g, 72 mmol ) and ethanol (23.2 g, 504 mmol). Chlorotrimethylsilane (54.8 g, 504 mmol) was then added dropwise with stirring at room temperature. The resulting solution was stirred for 4 hours at room temperature, then the reaction was stopped by adding 10 ml of water / ice. The resulting solution was extracted with 3x20 ml of petroleum ether (30-60 degree) and the organic layers were combined. The resulting mixture was washed with 2x20 ml brine. The mixture was dried over anhydrous sodium sulfate, filtered and concentrated. The residue was eluted with petroleum ether (30-60 degrees) on a silica gel column. This gave 6 g (45%) of the title compound as a colorless oil.

1H NMR (300 MHz, d6-DMSO): δ 6.57 (d, 1H, J = 56 Hz), 4.37 (q, J = 7.2 Hz, 2H), 1.33 (t, 3H, J = 7.2

Hz).

Intermediate 175. (E) -but-2-Enylboronic acid one

CH OH

- 99 037916

To a solution of (E) -but-2-en-1-ol (59.5 g, 826 mmol) in MeOH (360 ml) at room temperature was added H ₂ PdCl ₄ (2.06 g, 8.34 mmol), _{then B 2} (OH) ₄ (81.8 g, 919 mmol) was added to the mixture in portions at

30-40 ° C. The resulting solution was stirred at 30-40 ° C for 20 min, then filtered through celite.

¹ H-NMR (DMSO-d6, 400 MHz): δ 5.39-5.49 (m, 1H), 5.16-5.26 (m, 1H), 3.38 (s, 2H), 1 , 38-1.58 (m, 5H).

Intermediate 176. (S, E) -2- (m-t-Butylsulfinylimino) acetic acid

To molecular sieves type 4A (500 g) in DCM (600 ml) were added (S) 2-methylpropane-2-sulfinamide (100 g, 819 mmol) and 2-oxoacetic acid hydrate (91.2 g, 991 mmol ). The resulting solution was stirred at room temperature overnight, then filtered through celite.

¹ H-NMR (DMSO-d ₆ , 400 MHz): δ 7.77 (s, 1H), 5.74 (s, 1H), 1.13 (s, 9H).

Intermediate 177. (2S, 3R) -2 - ((S) -1,1-Dimethylethylsulfinamido) -3-methylpent-4enoic acid

A solution of (E) -but-2-enylboronic acid ( Intermediate 175) in MeOH (360 ml). The resulting solution was stirred at 015 ° C for 1 hour. The molecular sieves were removed by filtration and washed with DCM. The filtrate was removed by vacuum distillation to give the crude product. H2O (600 ml), petroleum ether (240 ml) and methyl tert-butyl ether (120 ml) were added to the crude product, stirred at room temperature for 1 hour, then filtered and the solid was collected and dried in vacuo at 25 ° C to give the product (110 g, 58%) as a white solid.

¹ H-NMR (DMSO-d6, 400 MHz): δ 5.71-5.80 (m, 1H), 5.01-5.07 (m, 2H), 4.94 (d, J = 8Hz , 1H), 3.58-3.62 (m, 1H), 2.56-2.61 (m, 1H), 1.15 (s, 9H), 0.975 (d, J = 4 Hz, 3H) ...

Intermediate 178. (2S, 3R) -Methyl-2-amino-3-methylpent-4-enoate

To a solution of (2S, 3R) -2 - ((S) -1,1-dimethylethylsulfinamido) -3-methylpent-4-enoic acid (Intermediate 177, 44 g, 189 mmol) in MeOH (200 ml) was added dropwise SOCl ₂ (68.6 ml, 944 mmol) at 0 ° C. The reaction mixture was stirred at 0 ° C for 1 h, then warmed to 70 ° C and stirred overnight. The solvent was removed and the residue was diluted with water. _{NaHCO 3 was} added to the aqueous phase to neutralize the pH to about 8, extracted with DCM, dried over Na ₂ SO ₄ , filtered and concentrated to give 23 g of the crude product as a yellow liquid.

¹ H-NMR (DMSO-d ₆ , 400 MHz): δ 5.71-5.82 (m, 1H), 4.97-5.05 (m, 2H), 3.61 (s, 3H), 3.27 (d, J = 8 Hz, 1H), 2.33-2.45 (m, 1H), 1.79 (s, 2H), 0.96 (d, J = 8 Hz, 3H).

LCMS: retention time = 0.682, [M + H] + 144.2.

Intermediate 179. (2S, 3R) -Methyl-2- (allylamino) -3-methylpent-4-enoate

LiOH (4.248 g, 177 mmol ). The mixture was stirred at 0 ° C for 30 minutes. Then a solution of 4-bromobut-1-ene (17.5 g, 145 mmol) in DMF (15 ml) was added dropwise. The reaction mixture was stirred at -7 ° C for 20 min, then slowly warmed to room temperature and stirred overnight. The reaction was stopped by adding water, extracted with EtOAc. The organic layer was washed with water, brine, dried over Na ₂ SO ₄ , filtered and concentrated to give 28 g of the crude product as a yellow liquid.

¹ H-NMR (DMSO-d6, 400 MHz): δ 5.65-5.83 (m, 2H), 4.93-5.16 (m, 4H), 3.60 (s, 3H), 3 , 14-3.22 (m, 1H), 3.06 (d, J = 8 Hz, 1H), 2.93-3.01 (m, 1H), 2.34-2.41 (m, 1H ), 0.99 (d, J = 8 Hz, 3H)

LCMS: retention time = 0.461, m / z: 184 [M + H].

Intermediate 180. (2S, 3R) -Methyl-2- (allyl (tert-butoxycarbonyl) amino) -3methylpent-4-enoate

- 100 037916

(Boc) ₂ О (33, 4 g, 153 mmol). The reaction mixture was stirred at room temperature for 10 min, then warmed to 90 ° C and stirred overnight. The reaction mixture was concentrated. The crude product was purified by flash chromatography on silica gel with a gradient eluent of 0 to 6% EtOAc in petroleum ether. The pure fractions were evaporated to give the product (24 g, 56%) as a light yellow liquid.

1 H-NMR (DMSO-d ₆ , 400 MHz): δ 5.71-5.84 (m, 2H), 5.00-5.11 (m, 4H), 3.66-4.43 (m, 3H), 3.58 (s, 3H), 2.81 (s, 1H), 1.39 (s, 9H), 0.93 (d, J = 8 Hz, 3H). LCMS: retention time = 1.096, 184 [M-Boc + H].

Intermediate 181. (2S, 3R) -1-tert-Butyl-2-methyl-3-methyl-2,3-dihydropyridine 1,2 (6H) -dicarboxylate

To a solution of (2S, 3R) -methyl-2- (αlyl (tert-butoxycarbonyl) amino) -3-methylpent-4-enoate (Intermediate 180, 24 g, 84.8 mmol) in DCM (250 ml) in three batches the first generation Grubbs catalyst (886 mg 1.06 mmol) was added at 0 ° C and stirred for 4 hours. The first generation Grubbs catalyst (886 mg 1.06 mmol) was added again at 0 ° C in three batches, then heated to 25 ° WITH. The resulting solution was stirred at room temperature overnight. LCMS: retention time = 0.987, 156.2 [M-Boc + H], 295 [M + K].

Intermediate 182. (2S, 5R) -1-tert-Butyl-2-methyl-5- (tert-butoxycarbonyl (hydroxy) amino) -3-methyl-5,6-dihydropyridine-1,2 (2H) -dicarboxylate

To a solution of (2S, 3R) -1-tert-butyl-2-methyl-3-methyl-2,3-dihydropyridine-1,2 (6H) -dicarboxylate (Intermediate 181, 21.6 g, 84.8 mmol ) in DCM (250 ml), BocNHOH (16.9 g, 127 mmol), CuCl (0.419 g, 4.24 mmol) and pyridine (87 mg, 1.1 mmol) were added and degassed with oxygen. The resulting solution was stirred at room temperature for 44 hours under an oxygen atmosphere. The solid was removed by filtration. The filtrate was washed with water (3x200 ml) and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by flash chromatography on silica gel, eluted with 0-50% DCM in petroleum ether to give the product (29 g, 88%) as a brown oil.

1 H-NMR (CDCl ₃ , 400 MHz): δ 5.66 (d, J = 16 Hz, 1H), 5.31 (s, 1H), 4.90 (d, 1H), 4.51 (d, 1H), 4.11 (t, J = 20 Hz, 1H), 3.75 (s, 3H), 3.50-3.61 (m, 1H), 1.90 (d, J = 8 Hz, 3H), 1.43-1.51 (m, 18H) LCMS: retention time = 0.713, 279 [M-Boc + Na].

Intermediate 183. (2S, 5R) -1-tert-Butyl 2-methyl-5- (tert-butoxycarbonyl (tert-butyldimethylsilyloxy) amino) -3-methyl-5,6-dihydropyridine-1,2 (2H) -dicarboxylate

To a solution of (2S, 5R) -1-tert-butyl-2-methyl-5- (tert-butoxycarbonyl (hydroxy) amino) -3-methyl-5,6-dihydropyridine-1,2 (2H) -dicarboxylate (Intermediate 182 , 23 g 59.5 mmol) in DCM (180 ml) was added imidazole (8.09 g, 119 mmol). The resulting solution was stirred at room temperature for 10 min, then a solution of TBS-Cl (11.6 g, 77.4 mmol) in DCM (20 ml) was added dropwise at 0-5 ° C. The reaction mixture was stirred at 0 ° C for an additional 18 hours. The organic phase was washed with water and brine, dried over Na2SO4, filtered and concentrated. The crude product was purified by flash chromatography on silica gel, eluted with petroleum ether and DCM to give the product (20 g, 67%) as a brown oil.

¹ H-NMR (CDCl3, 400 MHz): δ 5.7 (s, 1H), 4.45-4.85 (m, 2H), 4.09 (d, J = 20 Hz, 1H), 3, 76 (s, 3H), 3.533.58 (m, 1H), 1.9 (s, 3H), 1.45-1.54 (m, 18H), 0.92 (d, J = 16 Hz, 9H ), 0.10 (d, J = 20 Hz, 6H).

LCMS: retention time = 1.467,523.55 [M + Na].

Intermediate 184. (2S, 5R) -Methyl-5- (tert-butyldimethylsilyloxyamino) -3-methyl1,2,5,6-tetrahydropyridine-2-carboxylate ⁰ I

HN ^^ OTBS Η

To a solution of (2S, 5R) -1-tert-butyl-2-methyl-5- (tert-butoxycarbonyl (tert-butyldimethylsilyloxy) amino) -3-methyl-5,6-dihydropyridine-1,2 (2H) -dicarboxylate (Intermediate 183, 20 g, 40 mmol) in DCM (200 ml) was added ZnBr ₂ (35.5 g 160 mmol) at 0 ° C. The resulting solution was stirred

- 101 037916 at room temperature overnight. The solid was removed by filtration. The filtrate was washed with saturated NaHCO ₃ solution to neutralize the pH to about 8, extracted with DCM, dried over Na 2 SO 4, filtered and concentrated. The crude product was purified by flash chromatography on silica gel, eluted with 0-1% MeOH in DCM to give the product (8.4 g, 70%) as a brown oil.

1H-NMR (CDCl ₃ , 400 MHz): δ 5.59-5.62 (m, 1H), 3.82 (s, 1H), 3.75 (s, 3H), 3.14-3.23 (m, 2H), 2.92-2.97 (m, 1H), 1.79 (s, 3H), 0.90 (s, 9H), 0.10 (s, 6H).

LCMS: retention time = 1.073, 301.2 [M + H].

Intermediate 185. Methyl- (2S, 5R) -6- [tert-butyl (dimethyl) silyl] oxy-3-methyl-7-oxo1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxylate about I

Ύ L-O OTBS

To a solution of (2S, 5R) -methyl-5- (tert-butyldimethylsilyloxyamino) -3-methyl-1,2,5,6-tetrahydropyridine-2-carboxylate (Intermediate 184, 42 g, 140 mmol) in MeCN (840 ml) was added DIEA (72.2 g 560 mmol) and degassed with nitrogen, then a solution of triphosgene (16.408 g, 56 mmol) in MeCN (120 ml) was added dropwise at 0 ± 5 ° C under nitrogen atmosphere. The resulting solution was stirred at room temperature under nitrogen atmosphere overnight. The reaction mixture was concentrated, then EtOAc was added and washed with 1N HCl. citric acid, saturated NaHCO ₃ solution and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by C-18 stationary phase flash chromatography eluting with 0-38% MeCN in H2O to give the product (22 g, 49%) as an orange solid.

1 H-NMR (CDCl ₃ , 400 MHz): δ 6.15 (t, J = 1.6 Hz, 1H), 4.412 (s, 1H), 3.807 (s, 3H), 3.610-3.629 (m, 1H) , 3.506 (d, J = 11 Hz, 1H), 3.290-3.322 (m, 1H), 1.722 (s, 3H), 0.962 (s, 9H), 0.195 (s, 3H), 0.172 (s, 3H).

LCMS: retention time = 1.585, 327.35 [M + H].

Intermediate 186. (2S, 3R) -2-amino-3-methylpent-4-enamide hydrochloride o =

Λ, χ ^ η ₂ ν nh ₂ hci

To a solution (Intermediate 177, 75.2 kg, 322.8 mol, 1.0 eq.) In THF (600 L) was added CDI (61.9 kg, 382.1 mol, 1.2 eq.) In batches at 20 ± 10 ° C. The mixture was stirred for 2 h at 20 ± 10 ° C. The mixture was cooled to -40 ± 5 ° C. NH3H2O (43.55 kg, 640.4 mol, 2.0 eq., 25 wt%) was added dropwise at -40 ± 5 ° C. The mixture was stirred for 10 min at -40 ± 5 ° C. After completion of the reaction, the mixture was warmed to -10 ~ 0 ° C, then concentrated in vacuo to about 3.0 original volumes. Added THF (2.0 vol.) And concentrated in vacuo to about 3.0 original volumes. THF was changed twice with EtOAc (2.0 vol.). EtOAc (6.0 vol.) Was added to the solution and cooled to 5 ± 5 ° C. MeSO3H (148.3 kg, 1543.0 mol, 2.4 eq.) Was added dropwise at <10 ° C and stirred for 1 h at 5 ± 5 ° C. The mixture was centrifuged and the resulting solid was washed twice with EtOAc (1.0 vol.). The mother liquor was collected to give the compound in solution in EtOAc, which was used directly in the next step. HCl (gas) was bubbled through the solution at 0 ± 5 ° C for 17 h (about 4 kg / h). After completion of the reaction, the mixture was centrifuged and the resulting solid was washed twice with EtOAc (1.0 vol.). The precipitate was dried in vacuo at 25 ± 5 ° C for at least 12 hours to give the title compound as a light yellow solid (98 kg, 90% total yield from two steps).

1H NMR (400 MHz, DMSO): δ 8.24 (s, 2H), 8.00 (s, 1H), 7.56 (s, 1H), 5.84-5.76 (m, 1H), 5.17-5.09 (m, 2H), 3.70-3.68 (m, 1H), 2.77-2.72 (m, 1H), 1.04 (d, J = 6.8 Hz, 3H).

Intermediates 187 and 188. Allyl ((2S, 3R) -1-amino-3-methyl-1-oxopent-4-en-2-yl) and tert-butyl allyl ((2S, 3R) -1-amino -3-methyl-1-oxopent-4-en-2-yl) carbamate

O = O Ξ

Λ X ^ X

H ₂ N and H ₂ N \

To a solution of LiOH (27.55 kg, 1147.9 mol, 2.0 eq.) In DMF (285 L) was added Intermediate 186 (95 kg, 564.67 mol, 1.0 eq.) In batches at 0 ± 5 ° C. The mixture was stirred for 0.5 h. Allyl bromide (76.76 kg, 634.5 mol, 1.1 eq.) Was added dropwise over 9 h at 0 ± 5 ° C. The reaction mixture was warmed to 20 ± 5 ° C and stirred for at least 8 hours. The reaction mixture was cooled to 10 ± 5 ° C and softened water (3.0 vol.) Was added. The mixture was extracted three times with DCM (3.0 vol.). The combined organics were washed with brine (2.0 vol.). The brine layer was extracted with DCM (3.0 vol.). The combined organic layers were concentrated at atmospheric pressure at <60 ° C to about 5.0 original volumes. To the solution was added tert-butyl alcohol (2.0 vol.) Was added and concentrated in vacuo at <60 ° C to about 5.0 original volumes. Concentration with tert-butyl alcohol (2.0 vol.) Was repeated one more time to a water content of <0.2% to give Prome-102 037916 compound 187. Tert-butyl alcohol (8.0 vol.) Was added to the concentrated solution.

Boc2O (138.07 kg, 633.3 mol, 1.1 eq.) Was added at 20 ± 5 ° C. The mixture was warmed to 70 ± 5 ° C and stirred for at least 15 hours.

After completion of the reaction, the mixture was cooled to 40 ± 5 ° C, then concentrated in vacuo at <60 ° C to about 5 original volumes. The concentrated mixture was cooled to 25 ± 5 ° C and softened water (5.0 vol.) Was added. The mixture was extracted twice with methyl tert-butyl ether (4.0 vol.). The combined organics were washed once with 0.5M HCl solution (2.0 vol.), Three times with brine (1.0 vol.) And concentrated in vacuo at <50 ° C to about 2.0 original volumes. N-heptane (1.0 vol) was added to the reactor and concentrated in vacuo at <50 ° C to about 2.0 original volumes. This procedure was repeated one more time. To the concentrated solution was added n-heptane (1.0 vol.), Cooled to -10 ± 5 ° C and stirred for at least 2 hours. The mixture was centrifuged and the resulting solid was washed with chilled n-heptane (0.5 vol. ). The residue was dried in vacuo at 25 ± 5 ° C for at least 12 hours to provide Intermediate 188 as a white solid (84.2 kg, 100% purity, 54.4% overall two-step yield).

¹ H NMR (400 MHz, DMSO): δ 7.42 (s, 1H), 6.95 (s, 1H), 5.78-5.70 (m, 2H), 5.11-4.98 ( m, 4H), 4.33 (d, J = 10.8 Hz, 1H), 3.92-3.78 (m, 2H), 2.70 (br, 1H), 1.41 (s, 9H ), 0.88 (d, J = 6.4 Hz, 3H). LC / MS (ES ⁺ ) m / z 169.2 (M + H).

Intermediates 189. (2S, 5R) -tert-Butyl-5- (tert-butoxycarbonyl (hydroxy) amino) -2carbamoyl-3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

he

To a solution of Intermediate 188 (81.0 kg, 301.8 mol, 1.0 eq.) In DCM (810 L) was added the first generation Grubbs catalyst (1.215 kg, 1.5 mol, 0.005 eq.) In three batches at 0 ± 5 ° C. The reaction mixture was stirred for 1 h at 0 ± 5 ° C, then heated to 25 ± 5 ° C and stirred for 1 h. The mixture was cooled to 0 ± 5 ° C and the catalyst (1.215 kg, 1.5 mol, 0.005 equiv.) In three batches at 0 ± 5 ° C. The mixture was warmed up to 25 ± 5 ° С and stirred for at least 8 hours. After completion of the reaction, CuCl (1.46 kg, 14.8 mol, 0.05 eq.), BocNHOH (59.94 kg, 450.7 mol, 1.5 eq.) And pyridine (0.31 kg, 3.9 mol, 0.013 eq.) At 25 ± 5 ° C. The mixture was stirred for at least 43 hours at 25 ± 5 ° C under an oxygen atmosphere. After completion of the reaction, a solution of EDTA ₂ Na (5.0 vol.) Was added to the reactor and stirred for at least 4 hours at 25 ± 5 ° C. The layers were separated and the aqueous layer was extracted twice with DCM (3.0 vol.). The organics were combined and performed under vacuum at <40 ° C to about 3.0 original volumes. Methyl tert-butyl ether (MTBE) (2.0 vol.) Was added to the reactor and concentrated in vacuo at <40 ° C to about 3.0 original volumes. This procedure was repeated one more time. MTBE (2.5 vol.), N-heptane (2.5 vol.) And softened water (5.0 vol.) Were added to the concentrated solution, and the mixture was suspended at 20 ± 5 ° C for at least 2 h. The mixture was centrifuged and the pellet was washed with MTBE / n-heptane (0.5 vol., 1: 1). The precipitate was dried in vacuo at 35 ± 5 ° C for at least 12 h to a water content of <1% to give the title compound as a light brown solid (70.3 kg, 98% purity, two steps total yield 61.3%). Ή NMR (400 MHz, DMSO): δ 8.93-8.78 (m, 1H), 7.49 (s, 1H), 7.07 (s, 1H), 5.57 (s, 1H), 4.62-4.41 (m, 2H), 3.76-3.50 (m, 2H), 1.79 (s, 3H), 1.43 (s, 1H), 1.40 (s, 9H). LC / MS (ES ⁺ ) m / z 272.2 (M + H)

Intermediates 190 and 191. tert-Butyl- (3R, 6S) -3 - ((tert-butoxycarbonyl) ((tert-butyldimethylsilyl) oxy) amino) -6-carbamoyl-5-methyl-3,6-dihydropyridine-1 (2H ) -carboxylate and (2S, 5R) -5- (tert-butyldimethylsilyloxyamino) -3-methyl-1,2,5,6-tetrahydropyridine-2-carboxamide

To a solution of Intermediate 189 (50 kg, 134.6 mol, 1.0 eq.) And imidazole (18.5 kg, 268.1 mol, 2.0 eq.) In DCM (500 L) was added TBS- Cl (30.5 kg, 202.0 mol, 1.5 eq.) In solution in DCM (1.5 vol.) At 0 ± 5 ° С for 4.5 h.The mixture was heated to 20 ± 5 ° С and stirred for at least 5 hours. After completion of the reaction, softened water (250 L, 5.0 vol) was added. The layers were separated and the aqueous layer was extracted with DCM (3.0 vol.). The combined organic layers were washed twice with softened water (3.0 vol.), Then concentrated by atmospheric distillation at <50 ° C to about 2.0 original volumes. DCM (200 L, 4.0 vol) was added to the solution and concentrated by atmospheric distillation at <50 ° C to about 2.0 original volumes, to a water content of <1%, to give Intermediate 190. To the solution was added with stirring DCM (750 L, 15.0 vol.) At 20 ± 5 ° C under nitrogen. _{ZnBr 2} (60.5 kg, 268.9 mol, 2.0 eq.) Was added to the solution and stirred for 6 h at 20 ± 5 ° C. _{ZnBr 2} (30.5 kg, 135.6 mol, 1.0 eq.) Was added to the reaction mixture every 6-8 hours until the reaction was complete (about 5 eq. ZnBr _{2 in} total). The reaction was stopped by adding a solution (18.0 vol.) NaHCO ₃ (113.0 kg, 1345.2 mol, 10.0 eq.) At a temperature of less than 20 ° C. The mixture was stirred for at least 1 h at 20 ± 5 ° C, then centrifuged and the centrifugate collected. The resulting solid precipitate was suspended with dichloromethane (5.0 vol.) At 20 ± 5 ° C for 1 h, then centrifuged. The centrifugate was combined with the previously collected centrifugate and the layers were separated. The aqueous layer was extracted twice with DCM (3.0 vol.). The combined organics were washed four times with softened water (4.0 vol.), Then concentrated under normal pressure at <50 ° C to about 2.0 original volumes. To the solution was added CH ₃ CN (2.0 vol.) And concentrated in vacuo at <50 ° C to about 4.0 original volumes to afford Intermediate 191 in solution. LC / MS: (ES +) m / z 282.2 (M + H).

Intermediate 192. (2S, 5R) -6- (ret-Butyldimethylsilyloxy) -3-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-2-carboxamide

DIEA (69.56 kg, 539.2 mol, 4 , 0 equiv.) At 20 ± 5 ° С. The mixture was cooled to 0 ± 5 ° C and triphosgene (13.07 kg, 44.0 mol, 0.33 eq.) In CH3CN (115.2 L, 3.0 vol.) Was added dropwise. The mixture was warmed to 25 ± 5 ° C and stirred for at least 8 h, then cooled to 10 ± 5 ° C and quenched by the addition of softened water (24.19 kg, 1343.9 mol, 10.0 eq.). The mixture was stirred for at least 1 hour, then concentrated in vacuo at <40 ° C to about 5.0 original volumes. The solution was cooled to 10 ± 5 ° C and MTBE (384 L, 10 vol.) And brine (4.0 vol.) Were added. The layers were separated and the organics washed with brine (4.0 vol.), And concentrated in vacuo at <40 ° C to about 2.0 original vol. MTBE was replaced with n-heptane (1.0 vol.) And the solid was suspended for 1 h at 20 ± 5 ° C. The mixture was centrifuged and the precipitate was washed twice with n-heptane (0.5 vol.). The residue was suspended in MTBE (67.5 L) for at least 3 h at 20 ± 5 ° C, then n-heptane (336 L) was added with stirring for at least 1 h. The mixture was centrifuged and the resulting precipitate was washed with -heptane (1.0 vol.). The precipitate was dried in vacuo at 30 ± 5 ° C for 12 h to give the title compound as a light brown solid (21.1 kg, 99.6% purity, 49.6% overall three-step yield).

1H NMR (400 MHz, DMSO): δ 7.80 (s, 1H), 7.33 (s, 1H), 6.06 (t, J = 2 Hz, 1H), 4.10 (s, 1H) , 3.71-3.69 (m, 1H), 3.65 (d, J = 10.8 Hz, 1H), 3.10 (dd, J = 10.8 Hz, 2.0 Hz, 1H) , 1.63 (s, 3H), 0.92 (s, 9H), 0.14 (d, J = 0.8 Hz, 1H). LC / MS (ES ⁺ ) m / z 312.2 (M + H).

Intermediate 193. (2S, 5R) -tert-butyl-5- (tert-butoxycarbonyl (hydroxy) amino) -2carbamoyl-3-methyl-5,6-dihydropyridine-1 (2H) -carboxylate

To a solution of Intermediate 192 (18.5 kg, 59.4 mol, 1.0 eq) in EtOAc (92.5 L, 5 vol) was added HF-Py (2.04 kg, 71.4 mol, 1 , 2 equiv., 70 wt.%) At 0 ± 5 ° С. The mixture was stirred for at least 10 h at 20 ± 5 ° C, then cooled to 0 ± 5 ° C and HF-Py (0.33 kg, 11.6 mol, 0.2 eq., 70 wt. %). The mixture was stirred for at least 3 h at 20 ± 5 ° C, then MTBE (27.8 L, 1.5 vol.) Was added and stirred for 3 h at 10 ± 5 ° C. The mixture was centrifuged and the precipitate was washed with EtOAc (0.5 vol.). The pellet was suspended in EtOAc (2.0 vol.) For at least 1 h at 20 ± 5 ° C, then centrifuged. The precipitate was slurried in EtOAc (0.5 vol) then dried in vacuo at 25 ± 5 ° C for 12 h to give the title compound as a yellow solid (11.0 kg, 99.5% purity, yield 91%). 1H NMR (400 MHz, DMSO): δ 9.56 (s, 1H), 7.79 (s, 1H), 7.32 (s, 1H), 6.10 (d, J = 3.2 Hz, 1H), 4.06 (s, 1H), 3.68-3.62 (m, 2H), 3.07 (d, J = 8.4Hz, 1H), 1.61 (s, 3H). LC / MS (ES ⁺ ) m / z 198.1 (M + H).

Intermediate 194. Ethyl (S, E) -2 - ((tert-butylsulfinyl) imino) acetate

To a solution of ethyl 2-oxoacetate (66 ml, 321 mmol, 50% in toluene) in DCM (1 L) at 0 ° C was added (S) -2-methylpropane-2-sulfinamide (30 g, 248 mmol) and molecular sieves (4A, 500 g). The resulting solution was stirred at room temperature for 18 hours. The molecular sieves were removed by filtration; the filtrate was concentrated by distillation in vacuo to give the crude product, which was purified by flash chromatography on silica gel (0-5% EtOAc in petroleum ether) to 104 037916 to give a colorless oil, 45 g, 88%.

¹ H NMR (400 MHz, CDCl ₃ ): δ 1.28 (s, 9H), 1.39 (t, J = 12 Hz, 3H), 4.38 (q, J = 12 Hz, 2H), 8 , 01 (s, 1H).

Intermediate 195. Ethyl (S) -1 - ((S) -tert-butylsulfinyl) -4-methyl-1,2,3,6-tetrahydropyridine-2-carboxylate

Bu-t

Isoprene (97.21 ml, 971, 91 mmol) followed by TMSOTf (97.42 ml, 416.54 mmol). The resulting solution was stirred at -78 ° C for 3 hours and slowly quenched the reaction at -78 ° C by adding phosphate buffer solution (pH 7.4, 1 L). After warming to room temperature, the mixture was extracted with DCM (3x500 ml). The combined organic extracts were washed with water (2x500 ml) and brine. The organic layer was dried over Na ₂ SO ₄ , filtered and evaporated to give 60 g of the crude product as a brown oil. The product was used in the next step without further purification.

1H NMR (400 MHz, d ₆ -DMSO): δ 1.08 (s, 9H), 1.18 (t, J = 12 Hz, 3H), 1.64 (q, J = 4 Hz, 3H), 3.59 (m, 2H), 4.11 (dq, J = 12.4 Hz, 2H), 4.30 (dd, J = 8.4 Hz, 1H), 5.39 (ddd, J = 4 , 8 4 Hz, 1H);

LCMS: (ES ⁺ ) [M + H] ⁺ = 274; HPLC retention time = 1.78 min.

Intermediate 196.1- (tert-Butyl) -2-ethyl- (S) -4-methyl-3,6-dihydropyridine-1,2 (2H) dicarboxylate

Vos ''

To a solution of crude (S) -ethyl-1 - ((S) -tert-butylsulfinyl) -4-methyl-1,2,3,6-tetrahydropyridine-2-carboxylate (Intermediate 195, 100 g) in MeOH (1 l) hydrogen chloride (100 ml, 4M in dioxane) was added at 0 ° C. The resulting solution was stirred at room temperature for 18 h. MeOH and HCl / dioxane were removed by vacuum distillation to give the crude product, which was dissolved in water (1 L) and extracted with EtOAc (3x500 ml). The pH of the aqueous solution was adjusted to 7 with solid NaHCO3. The aqueous solution was extracted with EtOAc until no more product was detected by LCMS. The organic phases were combined and dried over Na ₂ SO ₄ , filtered and concentrated to give the crude product (30 g, 177 mmol) as a light yellow oil. The oil was dissolved in THF (500 ml) and cooled with an ice water bath. To the cooled solution was added a solution of sodium bicarbonate (22.3 g, 265.5 mmol) in water (500 ml) followed by di-tert-butyl dicarbonate (57.8 g, 265.5 mmol). The resulting solution was stirred at room temperature for 18 hours. The two layers were separated. The aqueous layer was extracted with ethyl acetate. The combined organic layers were dried over Na ₂ SO ₄ , filtered and evaporated. The crude product was purified by silica gel flash chromatography (0-30% EtOAc in PE) to afford the title compound 47.5 g, 43% yield from Intermediate 194.

1H NMR (400 MHz, CDCl ₃ ): δ 1.24 (t, 3H), 1.50 (m, 9H), 1.71 (s, 3H), 2.46 (m, 2H), 3.73 (m, 1H), 4.10 (m, 3H), 4.95 (m, 1H); LCMS: (ES ⁺ ) [M + Na] ⁺ = 292; HPLC retention time = 1.71 min.

Intermediate 197. mret-Butyl- (S) -2-carbamoyl-4-methyl-3,6-dihydropyridine-1 (2H) carboxylate o

Vos

To a solution of 1- (tert-butyl) 2-ethyl- (S) -4-methyl-3,6-dihydropyridine-1,2 (2H) -dicarboxylate (Intermediate 196, 47.5 g, 176 mmol) in THF (1000 ml) and water (500 ml) at 0 ° C was added dropwise lithium hydroxide (1 M, 440 ml, 440 mmol). The reaction mixture was warmed to room temperature and stirred for 16 hours. The solvent was removed; the residue was diluted with water. The pH of the solution was adjusted to about 3 with an HCl solution (1N). The mixture was extracted with EtOAc (3x300 ml).

The organic layers were combined, washed with water and brine, dried over MgSO ₄ , filtered and concentrated to give a colorless oil, 40.3 g.

1H NMR (300 MHz, d6-DMSO): δ 1.38 (m, 9H), 1.64 (s, 3H), 2.53 (m, 2H), 3.68 (m, 3H), 4, 73 (m, 1H), 5.35 (dd, J = 3, 15Hz, 1H), 12.45 (s, 1H); LCMS: (ES ⁺ ) [M + Na] ⁺ = 264; HPLC retention time = 1.01 min.

Intermediate 198. mret-Butyl- (S) -2-carbamoyl-4-methyl-3,6-dihydropuridine-1 (2H) carboxylate

- 105 037916

To a solution of (S) -1- (tert-butoxycarbonyl) -4-methyl-1,2,3,6-tetrahydropyridine-2-carboxylic acid (Intermediate 197, 40.3 g, 167.2 mmol) in THF ( 500 ml) at 0 ° C was added Ν, Ν'-carbonyldiimidazole (32.5 g, 200.6 mmol) in portions. The crude material was stirred at 0 ° C. for 5 hours. Ammonium acetate (38.2 g, 502.9 mmol) was then added. The reaction mixture was stirred at room temperature for an additional 18 h, the reaction was quenched by the addition of water and the reaction mixture was extracted with EtOAc. The combined organic layers were washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by silica gel flash chromatography (0-30% EtOAc in PE) to give a white solid, 25 g, 62%.

1H NMR (400 MHz, d6-DMSO): δ 1.41 (s, 9H), 1.66 (s, 3H), 2.35 (s, 2H), 3.84 (m, 2H), 4, 66 (m, 1H), 5.35 (m, 1H), 6.96 (s, 1H), 7.19 (s, 1H); LCMS: (ES ⁺ ) [M + Na] ⁺ = 263; HPLC retention time = 0.86 min.

Intermediate 199. tert-Butyl- (3R, 6S) -3 - ((tert-butoxycarbonyl) (hydroxy) amino) 6-carbamoyl-4-methyl-3,6-dihydropyridine-1 (2H) -carboxylate o

Vos

To a solution of tert-butyl (S) -2-carbamoyl-4-methyl-3,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 198, 25 g, 104.1 mmol) in DCM (250 ml) was added BocNHOH (70.6 g, 530.9 mmol), CuCl (6.1 g, 62.5 mmol) and pyridine (106.9 mg, 1.3 mmol). The resulting solution was stirred at room temperature for 44 hours under an oxygen atmosphere. The solids were removed by filtration. The filtrate was washed with water (6x500 ml) and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by flash chromatography on silica gel (0-50% EtOAc in PE) to afford the title compound as a white solid, 40% yield. The starting material was isolated (10 g). This procedure was repeated three times to obtain a total of 15 g of product. LCMS: (ES ⁺ ) [M + Na] ⁺ = 394 (C ₁₇ H ₂₉ N ₃ O ₆ ).

Intermediate 200. tert-Butyl- (3R, 6S) -3 - ((tert-butoxycarbonyl) ((tert-butyldimethylsilyl) oxy) amino) -6-carbamoyl-4-methyl-3,6-dihydropyridine-1 (2H ) -carboxylate

Vos

To a solution of tert-butyl (3R, 6S) -3 - ((tert-butoxycarbonyl) (hydroxy) amino) -6-carbamoyl-4-methyl3,6-dihydropyridine-1 (2H) -carboxylate (Intermediate 199, 12 g, 32.3 mmol) in DCM (96 ml) at 0 ± 5 ° C was added imidazole (4.4 g, 64.6 mmol). The resulting solution was stirred at room temperature for 10 min, then TBS-Cl (4.8 g, 32.3 mmol) in DCM (10 mL) was added dropwise. The reaction mixture was stirred at 0 ° C for an additional 18 h, washed with water and brine, dried over Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by silica gel flash chromatography (0-20% EtOAc in PE) to afford the title compound as a white solid, 10 g, 63%. LCMS: (ES ⁺ ) 486 (C ₂₃ H ₄₃ N ₃ O ₆ Si).

Intermediate 201. (2S, 5R) -5 - (((tert-butyldimethylsilyl) oxy) amino) -4-methyl1,2,5,6-tetrahydropyridine-2-carboxamide o otbs Ν H

To a solution of tert-butyl- (3R, 6S) -3- [tert-butoxycarbonyl- [tert-butyl (dimethyl) silyl] oxy-amino] -6-carbamoyl-4-methyl-3,6-dihydro-2H-pyridine-1 α-carboxylate (Intermediate 200, 21.7 g, 44.68 mmol) in DCM (250 mL) at 0 ° C was added zinc bromide (40.24 g, 178.71 mmol). The resulting suspension was allowed to warm to room temperature and stirred for about 66 hours. The reaction mixture was cooled with an ice water bath to which a suspension of NaHCO ₃ (38.23 g, 10 eq.) In water (300 ml) was added. The resulting mixture was stirred for 1 hour. The solid was removed by filtration and washed 3-4 times with DCM until no more product was found in the wash solution. The two layers of filtrate were separated. The aqueous layer was extracted with DCM three times (until no more product was found in the aqueous solution). The combined DCM solution was concentrated to remove most of the solvent. The residue was partially dissolved in 10% MeOH in DCM and loaded onto a thin layer of silica gel and eluted with 10% MeOH in DCM. The filtrate was evaporated and dried in vacuo to give a solid yellow foam (9.9 g crude material, 77%). MS: 286 ES + (C ₁₃ H ₂₇ N ₃ O ₂ Si).

Intermediate 202. (2S, 5R) -6 - ((tert-Butyldimethylsilyl) oxy) -4-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-ene-2-carboxamide

- 106 037916

Into a clear solution of (3R, 6S) -3 - [[tert-butyl (dimethyl) silyl] oxyamino] -4-methyl-1,2,3,6-tetrahydropyridine-6-carboxamide (Intermediate 201, 7.66 g , 26.83 mmol) in MeCN (150 ml) and DCM (200 ml) at 0 ° C was added N, N'-diisopropylethylamine (19.11 ml, 107.34 mmol), followed by a solution of triphosgene (2.71 g , 9.12 mmol) in MeCN (50 ml) dropwise (2 ml / h using a syringe pump). After the addition, the solution was allowed to warm to room temperature and stirred overnight. The reaction mixture was concentrated to dryness. The resulting residue was diluted with EtOAc and washed with brine. The aqueous layer was extracted with EtOAc. The combined organic extracts were dried over MgSO ₄ , filtered and concentrated. The crude product was purified by silica gel chromatography (0-100 EtOAc / hexane) to afford the title compound as a white solid, 4.36 g, 52%. MS: 312 ES + (C14H25N3O3S1).

Intermediate 203. (2S, 5R) -6-Hydroxy-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3ene-2-carboxamide

To a solution of (2S, 5R) -5 - (((tert-butyldimethylsilyl) oxy) amino) -4-methyl-1,2,5,6-tetrahydropyridine2-carboxamide (Intermediate 202, 165 mg, 0.53 mmol) in ethyl acetate (4 ml) at 0 ° C was added HF-pyridine (0.02 ml, 0.64 mmol). The reaction mixture was warmed to room temperature and stirred for 1 hour. Only a small amount of product was observed. Another equivalent of HF-pyridine was added and the reaction was stirred for 3 h. The reaction mixture was concentrated to give an orange solid. MS: 198 ES + (C ₈ H _n N ₃ O ₃ ).

Example 35. (2R) -Isopropyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3en-6-yl) oxy ) -2-fluoroacetate

To a solution of (2S, 5R) -6-hydroxy-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 203, 582 mg, 2.95 mmol) in 1,4-dioxane (16 ml) and DMF (2 ml) was added isopropyl- (2R) -2-bromo-2-fluoroacetate (Intermediate 169, 881.1 mg, 4.43 mmol). The reaction mixture was cooled to 0 ° C and DBU (0.44 mL, 2.95 mmol) was added dropwise. The reaction mixture was stirred for 10 min, then diluted with ethyl acetate and washed twice with a 1: 1 mixture of brine and water. The organics were dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0% -90% ethyl acetate / hexanes), a white foam was obtained. The foam was dissolved in a 1: 1 mixture of acetonitrile and water, frozen and lyophilized to give a white solid, 614 mg, 66%. 6% S-diastereomer was present. MS: 316 ES + (C13H18FN3O5).

1H NMR (300 MHz, DMSO-d6) δ: 1.22 (m, 6H); 1.81 (m, 3H); 3.17 (m, 1H); 3.34 (m, 1H); 3.93 (m, 1H); 4.22 (m, 1H); 5.01 (m, 2H); 5.51 (m, 1H); 6.23 (m, 1H); 7.31 (s, 1H); 7.55 (s, 1H).

Example 90. Ethyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) - 2,2-difluoroacetate

To a solution of (2S, 5R) -6-hydroxy-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 203, 64.5 mg, 0 , 33 mmol) and ethyl bromodifluoroacetate (0.13 ml, 0.98 mmol) in DMF (3 ml), potassium carbonate (135.62 mg, 0.98 mmol) was added at room temperature. The mixture was stirred for about 3 hours, then diluted with ethyl acetate and filtered. The filtrate was combined with the previous small batch, washed twice with a 1: 1 mixture of brine and water, dried over magnesium sulfate, filtered and concentrated. After chromatography on silica gel (0% -80% ethyl acetate / hexanes) and lyophilization, the title compound (52.1 mg, 34%) was obtained as an orange solid. MS: 320 ES + (C ₁₂ H ₁₅ F ₂ N ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d ₆ ) δ: 1.29 (t, 3H); 1.82 (m, 3H); 3.36 (m, 2H); 3.94 (m, 1H); 4.31 (m, 1H); 4.39 (m, 2H); 5.57 (m, 1H); 7.36 (s, 1H); 7.59 (s, 1H).

Example 91.2 - (((2S, 5R) -2-Carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) 2.2 -difluoroacetic acid lithium salt

- 107 037916

To a solution of ethyl-2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) -2 , 2-difluoroacetate (example 90, 46 mg, 0.14 mmol) in THF (2 ml) and water (0.50 ml) at 0 ° C was added 1M lithium hydroxide (0.14 ml, 0.14 mmol). The reaction mixture was stirred at 0 ° C for 10 min. Added another 0.2 eq. lithium hydroxide and after 5 min the reaction was neutralized by adding 0.5 N. hydrochloric acid, and the THF was evaporated. The resulting solution was frozen and lyophilized. After purification with a Gilson system (Synergi Polar RP 21.2 mm x 100 mm, 4 μm, coupled to YMC C30 20 mm x 150 mm, 5 μm, 0% -16% acetonitrile / water, 6 min), the title compound (27 , 2 mg, 64.8%) as an off-white solid. MS: 292 ES + (C ₁₀ H11F ₂ N ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 1.81 (m, 3H); 3.30 (m, 2H); 3.84 (m, 1H); 4.20 (m, 1H); 5.48 (m, 1H); 7.29 (s, 1H); 7.55 (s, 1H).

Example 92. Ethyl 2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-6yl) oxy) acetate

To a solution of (2S, 5R) -6 - ((tert-butyldimethylsilyl) oxy) -4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct3-ene-2-carboxamide (Intermediate 202, 152 mg, 0.49 mmol) in THF (4 ml) at 0 ° C was added TBAF (0.49 ml, 0.49 mmol). The mixture was stirred for about 10 minutes. Ethyl bromoacetate (0.05 ml, 0.49 mmol) was added to the reaction mixture and stirred for 10 minutes. Ethyl bromoacetate (0.05 ml, 0.49 mmol) was added again and stirred for 30 min. Ethyl bromoacetate (0.05 ml, 0.49 mmol) was added again and the reaction mixture was warmed to room temperature and stirred for 1 hour. The reaction mixture was concentrated on silica gel and purified (0% -90% ethyl acetate / hexanes) to give a colorless oil. The oil was dissolved in a 1: 1 mixture of acetonitrile and water, frozen and lyophilized to give the title compound as a white solid, 78.5 mg, 56%. MS: 284 ES + (C ₁₂ H ₁₇ N ₃ O ₅ ).

1H NMR (300 MHz, DMSO-d6) δ: 1.23 (t, 3H); 1.84 (m, 3H); 3.19 (m, 2H); 3.97 (m, 1H); 4.17 (m, 3H); 4.53 (m, 2H); 5.46 (m, 1H); 7.29 (s, 1H); 7.50 (s, 1H).

Example 93. 2 - (((2S, 5R) -2-Carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) acetic acid lithium salt

To a solution of ethyl 2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy) acetate ( example 92, 57.4 mg, 0.2 mmol) in THF (2 ml) and water (1 ml) at 0 ° C was added 1M lithium hydroxide (0.66 ml, 0.66 mmol). The reaction mixture was stirred at 0 ° C for 10 min. Added another 0.2 eq. lithium hydroxide was added and after 10 minutes the reaction was complete. The reaction mixture was neutralized by the addition of 0.5 N. hydrochloric acid and added 1 eq. sodium bicarbonate in water at 0 ° C. The resulting solution was frozen and lyophilized. After system purification from Gilson (0% -16%, 6 min), the title compound was obtained as a white solid, 18.3 mg, 35%. MS: 256 ES + (C10H13N3O5).

¹ H NMR (300 MHz, DMSO-d6) δ: 1.85 (m, 3H); 3.11 (m, 2H); 3.88 (m, 2H); 4.07 (m, 1H); 4.24 (m, 1H); 5.39 (m, 1H); 7.24 (s, 1h); 7.48 (s, 1H).

Intermediate 204. (2S, 5R) -6 - ((tert-Butyldimethylsilyl) oxy) -4-methyl-7-oxo-1,6 diazabicyclo [3.2.1] oct-3-ene-2-carbonitrile

The title compound was prepared from (2S, 5R) -6 - ((tert-butyldimethylsilyl) oxy) -4-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate compound 202, 0.205 g, 0.66 mmol) according to the procedure for Intermediate 139 to give the title compound (159 mg, 82%) as a white solid. MS: 294 ES + (C ₁₄ H ₂₃ N ₃ O ₂ Si).

Example 94. Ethyl- (2R) -2 - (((2S, 5R) -2-cyano-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl ) hydroxy) -2-fluoroacetate

- 108 037916

The title compound was prepared from (2S, 5R) -6 - ((tert-butyldimethylsilyl) oxy) -4-methyl7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carbonitrile (Intermediate compound 204, 152 mg, 0.49 mmol) according to the alternative procedure for Example 35 to give (26.9 mg, 19%) a colorless oil. Approximately 9% of the S-diastereomer was present. MS: 284 ES + (C ₁₂ H ₁₄ FN ₃ O ₄ ).

¹ H NMR (300 MHz, DMSO-d ₆ ) δ: 1.09 (t, 3H); 1.72 (m, 3H); 3.26 (m, 2H); 3.97 (m, 1H); 4.11 (m, 2H); 4.94 (m, 1H); 5.30 (m, 1H); 6.16 (m, 1H).

Example 95. (2R) -2 - (((2S, 5R) -2-Cyano-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6-yl) oxy ) -2fluoroacetic acid lithium salt

The title compound was prepared from ethyl- (2R) -2 - (((2S, 5R) -2-cyano-4-methyl-7-oxo-1,6diazabicyclo [3.2.1] oct-3-en-6- yl) oxy) -2-fluoroacetate (Example 94, 22 mg, 0.08 mmol) following the procedure for Example 91 to give (3.8 mg, 15%) a light yellow solid. MS: 256 ES + (C10H10FN3O4).

L NMR (300 MHz, DMSO-06) δ: 1.81 (m, 3H); 3.30 (m, 2H); 4.00 (m, 1H); 4.92 (m, 1H); 5.27 (m, 1H); 5.28 (m, 1H).

Example 96. Isopropyl 2 - (((2S, 5R) -2-carbamoyl-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-en-6yl) oxy) acetate

To a solution of (2S, 5R) -6-hydroxy-4-methyl-7-oxo-1,6-diazabicyclo [3.2.1] oct-3-ene-2-carboxamide (Intermediate 203, 93.38 mg, 0 , 47 mmol) and isopropyl bromoacetate (0.18 ml, 1.42 mmol) in DMF (4 ml), potassium carbonate (196.35 mg, 1.42 mmol) was added at room temperature. The reaction mixture was stirred for about 3 hours, then diluted with ethyl acetate and filtered. The filtrate was washed twice with a 1: 1 mixture of brine and water, dried over magnesium sulfate, filtered and concentrated. Chromatography on silica gel (0-80% ethyl acetate / hexanes) and lyophilization in acetonitrile afforded the title compound as an off-white solid, 106.6 mg, 72%. MS: 298 ES + (C13H19N3O5).

¹ H NMR (300 MHz, DMSO-BH) δ: 1.23 (m, 6H); 1.84 (m, 3H); 3.19 (m, 2H); 3.97 (m, 1H); 4.16 (m, 1H); 4.35 (m, 1H); 4.62 (m, 1H); 5.00 (m, 1H); 5.46 (m, 1H); 7.29 (s, 1H); 7.50 (s, 1H).

Biological examples

Example 102. Inhibition of beta-lactamase enzymes.

For all enzymes, a buffer consisting of 0.1 M sodium phosphate (pH 7.0), 10 mM NaHCO _3, and 0.005% Triton X-100 was used. The chromogenic substrate nitrocephin (SynGene, Bangalore, India) was used in an amount of 100 μM. The enzyme activity was monitored by the increase in absorbance at 490 nm as a result of nitrocefin hydrolysis. Assays were performed in clear 384-well polystyrene plates (Greiner Bio-One, Monroe, NC). Absorbance was measured over 1 hour at 30 second intervals using a Spectramax absorbance plate reader (Molecular Devices, Sunnyvale, Calif.). When measuring the inhibition of beta-lactamases with the INHIBITOR, serial 3-fold dilutions of the inhibitor in the assay buffer were used in the range from 100 μM to 62.7 pM. A background curve characterizing the uptake process for the control without enzyme and inhibitor was subtracted from each process curve. The complete set of process curves for a single enzyme with all inhibitor concentrations were numerically integrated using the Kintek Global Kinetic Explorer software (Kintek Corp, Snow Shue, PA) to best fit the mechanism shown in Scheme 10.

10 to 1 to _ scheme

E + S ES E + P ^to 1 ^to 2 to _e E + 1 5 ^ * _E | ^to s

109 037916 where E, S, ES, P, I and EI respectively represent the concentrations of the enzyme, nitrocefin, enzyme-nitrocefin complex, nitrocefin hydrolysis product, INHIBITOR and enzyme-INHIBITOR complex. The measured K _m (nitrocefin) value was used to determine the fixed values of k ₊₁ , k-1 and k ₊₂ , where k ₊₁ = k-1 = 1 and k ₊₂ = K _m -1. The values k ₊₂ , k ₊₃ and k _-3 were adjusted. To correct for minor differences in background absorbance between wells in absorbance measurements, a concentration shifted series was used. The parameter k _{+3 is} equivalent to the second-order rate constant k _mact / K1. In some cases, the inhibition was in rapid equilibrium over the time scale of the experiment, so that only the ratio k _-3 / k ₊₃ = K1 could be determined. Although k _-3 represents the reversal of inhibitor binding in Scheme 10, hydrolysis of the enzyme-inhibitor complex cannot be ruled out based on kinetic measurements.

The most closely related absorbance values obtained during the above procedure at each time point for each compound concentration were exported to Excel. To calculate the IC ₅₀ at 60 min,% inhibition was calculated for each inhibitor concentration at 60 min based on the most closely matched absorbance values at that time point using the equation:

% inhibition = 100x (1 - A _inhib / A _max ), where A is the most closely corresponding absorbance value without an inhibitor, and A _inhib is the most closely corresponding absorbance value in the presence of an inhibitor. IC _{50 was} calculated from a set of% inhibition values using nonlinear least squares regression using the equation:

% inhibition = 100 [I] ⁿ (IC ₅₀ + [I] ⁿ ), where [I] is the inhibitor concentration and n is Hill's coefficient. For nonlinear regression, the Excel XLfit add-in (Ш Business Solutions) was used. Table 1 shows the IC50 values of the compound samples (μM).

Table 1

Example Class A TEM-1 60 min IC50 (μM) Class C AshrC 60 min IC50 (μM) Class D OXA-48 60 min IC50 (μM) 3 0.019 0.071 0.015 4 0.00135 0.011 0.015 five 0.0044 0.02 0.017 eight 0.0015 0.012 0.014 10 0.43 1,2 0.051 AND 0.23 0,4 22 27 0.047 0.0083 0.018 thirty 0.081 0.0018 0.14 33 0.0032 0.16 0.077 34 0.026 oh, h 0.019

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41 0.0035 0.022 0.0064 42 0.024 0.095 0.0052 55 0.024 0.18 0.0073 56 0.093 0.14 0.22 57 0.057 0.0041 0.41 62 0.21 0.41 0.87 63 oh and 0.04 0.23 66 0.66 0.054 0.012 67 0.064 0.0073 0.34 68 0.31 0.0093 0.078 69 0.18 0.002 oh and 70 0.026 0.13 0.087 71 oh and 0.0044 0.075 72 0.077 0.053 oh and 73 0.73 0.01 0.53 74 0.28 0.022 0.59 75 0.083 0.014 0.42 76 0.94 44 0.22 77 > 100 > 100 > 100 78 3.6 > 100 0.69 79 4.2 > 100 5.7 82 0.085 0.014 0.028 87 0.15 0.017 0.011 88 0.054 0.0031 0.045 89 0.033 0.0021 0.15 91 0.00047 0.036 0.018 93 0.15 4.6 0.053 95 0.035 0.59 0.0013 o ΗζΝ ^ 'γ ^ M F Λ --- Ν _κ ? ⁰ ° ^ he o Comparative Compound 98 0.0013 2 2.6 HN ^- ^ O U. ⁰ ° ^ y-one O Comp. 99 0.03 1.6 82

Example 103. Recovery of the activity of cefpodoxime in the presence of a fixed concentration of 4 μg / ml of a beta-lactamase inhibitor.

Minimum inhibitory concentration (MIC) values for each organism and drug combination were determined using the broth microdilution technique from the Clinical and Laboratory Standards Institute (CLSI) Manual (CLSI M07-A10). Recommended quality control (QC) bacterial strains E. coli ATCC 25922, E. coli ATCC 35218 and Klebsiella pneumoniae ATCC 700603 were included in each of the tests in accordance with the ICLS guidelines to ensure there is no variation between test dates (CLSI M100-S25) ... The MICs of these QC strains were within the QC range in all tests performed. Plates containing the drug were made using the replica method. Cefpodoxime solution with 20-fold concentration and serial 2-fold dilutions were made in Mueller-Hinton broth with standardized cation content. Equal volumes of compound samples at a fixed 20-fold concentration were added to the original plate. 10 μl were imprinted on daughter plates using a Tecan EVO robot. Suspensions of organisms were adjusted to a McFarland 0.5 standard and further diluted to give 3x10 ⁵ to 7x10 ⁵ colony forming units (CFU) / ml of finished inoculum. Bacterial inoculum was prepared in sterile cation-standardized Mueller-Hinton broth (Beckton Dickinson). Inoculum in a volume of 90 μl at 1.1-fold concentration was added to the wells (using a Tecan EVO robot). All seeded microdilution plates were incubated in ambient air at 35 ° C for 18-24 hours. After incubation, the lowest drug concentration that prevented visible growth, as determined by optical density at 600 nm, was recorded as a MIC (Table 2, all MICs are indicated in μg / ml and all beta-lactamase inhibitors were tested at a fixed concentration of 4 μg / ml).

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table 2

Compound C. freundui E. coli Klebsiella pneumoniae Beta-lactamase content: AmpC, TEM-1, CMY65 Wild type (ATCC 25922) AmpC, OXA-1, STX-M15, TEM-1 SHV-18, OXA-1, OKR-6 (ATCC 700603) KRS-2, SHV-11, TEM-1 OHA-9 Cefpodoxime alone > 64 one > 64 sixteen > 64 + 3 <1 <0.5 one 2 0.5 + 4 <0.125 <0.06 0.06 0.25 0.06 + 5 <0.03125 <0.03125 <0.03125 0.0625 <0.03125 + 10 2 0.5 4 2 one + 11 sixteen 0.5 2 2 4 + 27 <0.03125 <0.03125 <0.03125 0.25 <0.03125 + 28 32 one 4 one 2 + 30 0.5 0.03125 0.5 2 one + 33 <0.03125 <0.03125 <0.03125 <0.0625 <0.03125 + 34 <0.03125 <0.03125 <0.03125 <0.125 <0.0625 + 41 <0.00012 <0.00012 0.00024 0.25 0.01117 + 42 4 0.5 2 2 one + 43 0.13 0.016 0.0039 one 0.063 + 55 > 32 0.0625 2 2 32 + 56 eight <0.03125 <0.03125 0.125 0.0625 + 57 <0.0625 <0.0625 <0.0625 one 0.0625 + 62 <0.03125 <0.03125 <0.03125 <0.0625 <0.03125 + 63 > 32 0.25 32 eight > 32 + 66 sixteen 0.5 > 32 4 eight + 67 0.5 <0.03125 <0.03125 4 one + 68 > 32 0.5 > 32 4 eight + 69 > 32 0.25 sixteen 4 eight + 70 > 32 0.5 sixteen 4 > 32 + 71 0.25 0.5 2 2 one + 72 > 32 <0.03125 4 4 eight + 73 > 32 0.25 32 BUT sixteen + 74 0.5 <0.03125 <0.03125 2 0.03125 + 75 <0.03125 <0.03125 <0.03125 0.125 <0.03125 + 76 > 32 one > 32 sixteen > 32 + 77 <0.03125 <0.03125 <0.03125 <0.03125 <0.03125 + 78 > 32 one > 32 eight > 32 + 79 > 32 one > 32 sixteen > 32 + 82 <0.03125 <0.03125 <0.03125 0.0625 <0.03125 + 87 <0.03125 <0.03125 <0.03125 0.125 <0.03125 + 88 <0.03125 <0.03125 <0.03125 2 <0.03125 + 89 <0.03125 <0.03125 <0.03125 0.0625 <0.03125 + 91 <0.03 <0.03 <0.03 <0.03 <0.03 + 93 <0.03 <0.03 <0.03 0.125 <0.03 + 95 32 0.25 one 2 2 O η ₂ ν · Ά |> M F X - '/ o ° y- ° ^H + o Comparative Compound 97 > 32 one 2 2 2 O n ₂ A · ^ M F J - N _x F ⁰ ° ^ yon + o Comp. 98 sixteen 0.5 2 2 0.5 ΗΝ'Χ Oh A ”” 'o ^ yon + o Comparative compound 99 > 32 0.5 eight 2 4

Example 104. Recovery of the activity of various oral beta-lactams in the presence of a fixed concentration of 4 μg / ml of a beta-lactamase inhibitor.

According to the procedure used in example 103, the MIC was determined for several beta-lactams in the presence of a fixed concentration (4 μg / ml) of samples of the compounds (Table 3) in relation to 3 strains of Enterobacteriaceae.

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Table 3

Compound C. freundui E. coli Klebsiella pneumoniae AmpC, TEM-1, CMY65 Wild type (ATCC 25922) SHV-18, OXA-2, OKR-6 (ATCC 700603) Cefpodoxime > 64 one sixteen Cefpodoxime + Pr. 3 (4 μg / ml) 0.5 0.125 2 Cefpodoxime + Pr. 4 (4 μg / ml) <0.125 <0.06 0.25 Cefpodoxime + Pr. 33 (4 μg / ml) <0.03125 <0.03125 0.25 Cefpodoxime + Pr. 34 (4 μg / ml) <0.03125 <0.03125 0.25 Cefpodoxime + Pr. 93 (4 μg / ml) <0.06 <0.06 0.25 Cefuroxime > 64 4 32 Cefuroxime + Pr. 3 (4 μg / ml) one 0.02 but Cefuroxime + Pr. 4 (4 μg / ml) 0.02 0.02 but Cefuroxime + Pr. 33 (4 μg / ml) <0.03125 <0.03125 4 Cefuroxime + Pr. 34 (4 μg / ml) <0.03125 <0.03125 one Tigemon 4 one 32 Tigemonam + Ave. 3 (4 μg / ml) 0.125 one 2 Tigemonam + Ave. 4 (4 μg / ml) <0.06 <0.06 0.5 Tigemonam + Ave. 33 (4 μg / ml) <0.06 <0.06 0.5

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Tigemonam + Ave. 34 (4 μg / ml) <0.06 <0.06 0.5 Tigemonam + Ave. 93 (4 μg / ml) 0.125 0.125 one Teipenem 0.125 <0.06 0.125 Teipenem + Ave. 3 (4 μg / ml) <0.06 <0.06 0.125 Teipenem + Ave. 4 (4 μg / ml) <0.06 <0.06 <0.06 Teipenem + Ave. 33 (4 μg / ml) <0.06 <0.06 0.125 Teipenem + Ave. 34 (4 μg / ml) <0.06 <0.06 <0.06 Teipenem + Ave. 93 (4 μg / ml) <0.06 <0.06 <0.06 Faropenem eight 0.5 eight Faropenem + Ave. 3 (4 μg / ml) 0.25 0.25 2 Faropenem + Ave. 4 (4 μg / ml) <0.06 <0.06 one Faropenem + Ave. 33 (4 μg / ml) <0.06 <0.06 0.5 Faropenem + Ave. 34 (4 μg / ml) <0.06 <0.06 0.5 Faropenem + Ave. 93 (4 μg / ml) <0.06 <0.06 one Cefixime > 64 0.5 eight Cefixim + Pr. 3 (4 μg / ml) one 0.125 0.5 Cefixim + Pr. 4 (4 μg / ml) <0.06 <0.06 <0.06 Cefixim + Pr. 33 (4 μg / ml) <0.06 <0.06 <0.06 Cefixim + Pr. 34 (4 μg / ml) <0.06 <0.06 <0.06 Cefixim + Pr. 93 (4 μg / ml) <0.06 <0.06 0.125 Loracarbef > 64 2 32 Loracarbef + Ave. 3 (4 μg / ml) one 0.5 0.5 Loracarbef + Ave. 4 (4 μg / ml) <0.06 <0.06 0.125 Loracarbef + Ave. 33 (4 μg / ml) <0.06 <0.06 0.125 Loracarbef + Ave. 34 (4 μg / ml) <0.06 <0.06 0.125 Loracarbef + Ave. 93 (4 μg / ml) <0.06 <0.06 0.25

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Example 105 Stability / conversion in the absence or presence of metabolizing enzymes.

Preparations of human intestinal S9 fraction in the absence of phenylmethylsulfonyl fluoride (PMSF), human liver S9 fraction, rat intestinal S9 fraction, and rat liver S9 fraction were obtained from Xenotech (Lenexa, Kansas).

The 500 μl incubation solution contained 0.8 mg / ml enzyme (or did not contain enzyme for stability studies in buffer), 10 μM test compounds in 100 mM HEPES buffer, pH 7.4. Hydrolysis reactions were performed in a 1 ml glass insert (Analytical Sales, Pompton Plains, NJ) in a shaking water bath at 37 ° C. After 0, 2, 5, 10, 15, 30 and 60 min, the reaction was stopped by pipetting 50 μl of the incubate into a DeepWell 96-well plate (Thermo Fisher Scientific, Rochester, NY) containing 100 μl of acetonitrile with 250 ng / ml carbutamide (Sigma-Aldrich, St. Louis, MO) as internal standard. The precipitated solutions were then vortexed well and then centrifuged at 4000 rpm for 15 min at 4 ° C. The extract was transferred to a new 96-well DeepWell plate for analysis by LC-ms / ms.

LC-MS / MS analysis was performed using an AB Sciex Qtrap 6500 mass spectrometer in positive ionization mode coupled to a Schimadzu Nexera LC system.

A Waters Atlantis T3 column (3 μm, 3.0x50 mm) was used for separation.

The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B) at a flow rate of 1.2 ml / min. For the prodrug assay, compound-specific multiple reaction monitoring (MMP) was set up so that both the prodrug and its hydrolyzed active ingredient could be monitored simultaneously.

Table 4 lists the first order half-lives of the compound samples (T _1/2 in mines). Half-lives quoted are based on the disappearance of the parent ester. As you can see from the table. 4, there is a good correlation between rat and human enzymes. Data in table. 4 indicate that, in general, there is a more rapid conversion of the ester to the active compound by the enzymes of the human and rat liver S9 fraction, while in the presence of buffer and intestinal S9 fraction, the esters are predominantly stable.

Table 4

Example Buffer, pH 7.4 Rat intestine S9 fraction Rat liver S9 fraction Human intestinal S9 fraction Human liver S9 fraction one 41 39 4.4 38 6.5 2 29 24 1.4 21 1.9 7 3.9 3.9 0.5 2.4 0.8 nine 18.3 27.3 1.4 17.8 2.9 12 236 233 fourteen 132 sixteen 13 55 but BUT 44 one fifteen 98 > 100 1.5 > 100 12.4 18 18 sixteen 7 fifteen 9.2 twenty 39 54 6.1 40 5.1 22 38 56 9.2 43 4.6 24 > 100 BUT BUT 34 nine 31 44 42 twenty 35 27 32 22 18 2.4 6.9 2.4 35 186 240 32 163 39 36 91 68 3 36 8.5 45 33 17 0.22 2 0.39 47 12 eleven 1.8 8.2 2,3 48 62 fifty 3 42 6.1 49 68 26 1.8 32 6.9 fifty 120 thirty 2.8 73 3.5 51 75 4.1 1.7 1.5 6.4 52 76 39 0.45 5.7 0.84 53 4.8 4.2 2 <1.0 1.7 54 22 nineteen 0.9 five 1.9 60 29 21 10 22 9.9 64 1,2 1.1 <1.0 <1.0 <1.0 65 1.7 1,3 <1.0 <1.0 <1.0 85 26 21 3.8 twenty 3.5 86 39 43 23 36 24 90 2.9 3.3 0.7 one 0.7 92 160.9 397 eleven 193.1 24.8 94 46.9 25.9 1,3 10.6 0 96 > 100 210.7 10.3 1475.6 57

H / O = undefined.

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Example 106. Pharmacokinetics in rats.

Pharmacokinetics in rats after intravenous administration of samples of compounds was determined in Sprague-Dawley rats (n = 3) with cannulated jugular vein (Harlan Laboratories, Indianapolis, Indiana) at a dose of 10 mg / kg. The compounds were dissolved and injected intravenously in 0.9% saline solution, pH 6.5 at a dose of 2 ml / kg. Blood samples (about 100 μl) were obtained through a catheter in the jugular vein before injection and at 0.08, 0.17, 0.25, 0.5, 1, 2, 4 and 8 hours after injection and prepared for plasma isolation in tubes Microtainer with K ₂ EDTA. Oral pharmacokinetic studies have been conducted with compound samples at 10 mg / kg carboxylic acid equivalents. Doses were dissolved and administered orally in a mixture of PEG400 and water for injection in a ratio of 25:75, pH 4.5 at a dose volume of 10 ml / kg. Blood samples were obtained through a jugular vein catheter prior to administration and at 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration and prepared for plasma isolation in Microtainer tubes with K2EDTA. Plasma samples were stored at -80 ° C prior to biological analysis.

Prepare samples for LC / MS / MS analysis.

Plasma samples were thawed on ice prior to processing. Samples (30 μl) were diluted and proteins precipitated with 180 μl of acetonitrile containing 0.1% formic acid and 250 ng / ml carbutamide (N1- (butylcarbamoyl) sulfanilamide, cat. # Sigma-Aldrich S385433) as an internal standard. The samples were vortexed for 30 s, centrifuged at 3400g for 10 min, and the supernatant was transferred to injection vials.

LC / MS / MS conditions.

Rapid hydrolysis prevented satisfactory analysis of the circulating concentrations of Examples 12 and 35. LC / MS / MS analysis was performed using an AB Sciex QTrap 6500 mass spectrometer in positive ionization mode coupled to a Schimadzu Nexera LC system.

A Waters Atlantis T3 column (3 μm, 3.0x50 mm) was used for chromatographic separation. The volume of the injected sample was 1 μL. The mobile phase consisted of 0.1% formic acid in water (A) and 0.1% formic acid in acetonitrile (B) with a gradient of 2-98% B: A for two minutes.

Pharmacokinetic analysis.

Plasma concentration versus time profiles after intravenous (iv) and oral (po) administration of compound samples were analyzed by non-compartmental analysis using WinNonLin 6.4. Average values of AUC and F% are shown in table. 5. The absolute oral bioavailability (F%) of the analyzed compounds after administration of their respective esters was determined as:

F% = 100 * (AUC _{π /} o * Dose / in) / (AUC _in . / In ^^^).

In general, compound samples show an increased oral AUC compared to sulfate-derived beta-lactamase inhibitors, resulting in a favorable F%.

Table 5

Introductory Example Method of administration Analyzed compound AUC, μM * h F% 4 i / v 4 19.1 3 i / v 3 34.9 13 on 3 18.3 53 12 on 4 8.2 38 2 on 3 18.5 58 one on 4 8.4 45 60 on 62 2.2 6.3 33 i / v 33 26.3 34 i / v 34 16.1 36 on 34 13.5 84 35 on 33 25.8 98 32 on 34 H, 5 71 31 on 33 19.8 75 53 on 4 2.2 eleven 65 on 4 2.8 fifteen nLA m J О OSO ₃ H ЕТХ2514 on ЕТХ2514 0.44 ηνΎ about BUT About OSO3H Relebactam on Relebactam 0.29 Hell J - N About OSO3H WCK 4234 on WCK 4234 0.45

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Example 106 Oral in vivo efficacy of cefpodoxime proxetil in combination with example 35 against E. coli (beta-lactamase content: AmpC, CTX-M-14).

The in vivo oral efficacy of Example 35 was evaluated in combination with cefpodoxime proxetil in a murine model of neutropenic femoral infection against the corresponding clinical strain. The strain, E. coli ARC2687, expresses the beta-lactamases AmpC and CTX-M-14, both of which can easily hydrolyze cefpodoxime, resulting in insensitive MICs exceeding 512 μg / ml. In combination with example 33 (4 μg / ml) MIC of cefpodoxime is reduced to <0.03 μg / ml. The study dose was adjusted to achieve an exposure of cefpodoxime above the MIC of 0.03 μg / ml over at least 50% of the dosing interval for all treatment groups with progressively increasing doses of the compound of Example 35. CD-I mice (Charles River, Wilmington, USA) were kept in shoebox type cages with contact mats and allowed to acclimate to containment for a minimum of 2 days prior to use. The animal room was maintained at 70 ° F, 50 ± 10% relative humidity, and a 12 hour light / dark cycle. The study was conducted using a protocol approved by the IACUC (Committee on the Care and Use of Laboratory Animals) in accordance with Title 9 CFR. Animals were induced neutropenia with two intraperitoneal doses of cyclophosphamide (150 mg / kg 4 days and 100 mg / kg 1 day before infection (Gerber et al. (1983) J Infect Dis 147 (5); 910917)). The animals were infected by intramuscular injection of about 1 x ¹ x 10 6 CFU in 100 μl of 0.9% saline. Inoculum was prepared from 25 ml of an overnight culture of E. coli ARC 2687 in trypticase-soy broth medium. After determining the optical density at 600 nm, the inoculum was diluted in 0.9% saline to a concentration of about 1.0x10 ⁶ CFU / ml before inoculation in the left and right thighs. Oral therapy with cefpodoxime with proxetil alone and in combination with Example 35 was started 2 hours after the bacterial challenge. Doses were suspended in 0.5% HPMC / 0.1% Tween 80 and administered orally via a gastric tube at a dose of 10 ml / kg. The endpoint was reached after 24 hours for determining the number of bacteria in the tissue of the thigh (CFU / g). The animals were euthanized in an ethical manner, the thighs were aseptically removed, weighed and homogenized in 1 ml of saline. The calculation of the bacterial load of the tissue homogenate was carried out by serial dilution in trypticase-soy agar plates, which were incubated overnight at 37 ° C until the colony count (CFU). The lower limit of detection was about 2.6 log ₁₀ CFU / g tissue.

As shown in table. 6, the bacterial load in the thighs of mice treated with cefpodoxime proxetil alone at a dose of 50 mg / kg every 6 hours or Example 35 at a dose of 10 mg / kg every 6 hours showed an increase of more than 3 log ₁₀ CFU / g after 24 hours of therapy. In combination with cefpodoxime proxetil, increasing the dose of the compound of Example 35 resulted in a dose-dependent decrease in bacterial load, with the maximum logarithmic reduction of -0.75 log ₁₀ CFU / g (relative to the initiation of therapy) was achieved at 50 mg / kg of cefpodoxime proxetil + 100 mg / kg of Example 35 every 6 hours. Meropenem, used as a positive control, provided a log reduction of CFU just above 1-log ₁₀ relative to the initiation of therapy at a dose of 600 mg / kg every 6 hours, administered subcutaneously.

Table 6

Group Dose (mg / kg) Method / Mode logioKOE / femoral tissue SC O Change of EKOE after 2 hours ECOE change after 26 hours Start therapy not apply MO not applicable 6.27 0.35 -4.59 26 h growth control carrier n / a / every 6 hours 10.86 0.19 +4.59 - Cefpodoxime a proxetil alone fifty n / a / every 6 hours 10.24 0.15 +3.97 -0.62 Example 35 on detached 10 n / a / every 6 hours 9.56 0.29 +3.29 -1.30 Example 35 + cefpodoxime a proxetil 10 + 50 n / a / every 6 hours 5.77 0.14 -0.50 -5.09

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Example 35 + cefpodoxime a proxetil 25 + 50 n / a / every 6 hours 5.59 0.17 -0.68 -5.27 Example 35 + cefpodoxime a proxetil 100 + 50 n / a / every 6 hours 5.52 0.20 -0.75 -5.34 Meropenem 600 n / c / every 6 hours 5.17 0.27 -1.10 -5.69

CLAIM

Claims (12)

CLAIM 1) phenyl optionally substituted with C ₁ -C ₃ -α-alkyl or C ₁ -C ₃ -alkyl-NH ₂ , 1. Compound of formula (II) or (III) or a pharmaceutically acceptable salt thereof, wherein R ^{1 is} selected from —CN, —CH2 NH2, —C (O) NH ₂ , where R ¹¹ is hydrogen or —C (O) NH ₂ , or R ^{1 is} selected from —C (O) NR ⁷ R ⁸ , where R ⁷ is hydrogen and R ⁸ is: 2. A compound according to claim 1, wherein R ¹ is —CONH2. 2) C ₁ -C ₃ -alkyl or 3. The compound according to claim 1 of formula (V) or a pharmaceutically acceptable salt thereof. 3) C ₁ -C ₃ alkoxy, where each alkyl or alkoxy of R ^{8 is} optionally and independently substituted with C ₃ -C ₆ cycloalkyl, -CN, -OH, -NH ₂ , -SO2NH2, -NHSO2NH2, -C (O ) NH ₂ , -NHC (O) (C ₁ -C ₃ -αalkyl), pyrazinyl, oxytanyl, oxazolyl or pyrrolidinyl optionally substituted with one or more carboxyl, fluorine or -C (O) O (C ₁ -C ₆ -alkyl ); R ² is C ₁ -C ₃ -alkyl; R ³ is C ₁ -C ₃ -alkyl; R ⁴ and R ⁵ independently represent hydrogen, methyl or fluoro; and R ⁶ is hydrogen, C1-C ₁₂ -alkyl, C ₁ -C ₄ -alkyl-OC (O) - (NHC ₁ -C ₆ -alkyl) -C (O) C ₁ -C ₃ alkoxy, C ₁ - C ₄ -alkyl-OC (O) -C ₁ -C ₄ -alkyl or C ₁ -C ₄ -alkyl-OC (O) -C ₁ -C ₃ -alkoxy. 4. A compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein R ⁶ is C ₁ -C ₁₂ alkyl. 5. A compound according to claim 4, or a pharmaceutically acceptable salt thereof, wherein R ⁶ is isopropyl. 6. A compound according to any one of claims 1-5, or a pharmaceutically acceptable salt thereof, wherein one of R ⁴ and R ⁵ is hydrogen and the other is fluorine. 7. A compound represented by one of the following formulas, or a pharmaceutically acceptable salt thereof: - 118 037916 - 119 037916 - 120 037916 - 121 037916 - 122 037916 123 037916 - 124 037916 - 125 037916 8. Compound of formula or a pharmaceutically acceptable salt thereof. - 126 037916 9. Compound of formula or a pharmaceutically acceptable salt thereof. 10. The use of a compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, in combination with a beta-lactam antibiotic in the treatment of a bacterial infection in a subject in need thereof. 11. Use according to claim 10, wherein said beta-lactam antibiotic is cefpodoxime proxetil or cefuroxime axetil. 12. Use according to claim 10 or 11, wherein the bacterial infection is selected from the group consisting of complicated urinary tract infection, uncomplicated urinary tract infection, kidney infection, lower respiratory tract infection, nosocomial bacterial pneumonia (NBP), pneumonia, acute bacterial prostatitis, acute bacterial infection of the skin and soft tissues, sepsis, intra-abdominal infection and infection associated with diabetic foot.