WO2013052568A1

WO2013052568A1 - Antibacterial compounds

Info

Publication number: WO2013052568A1
Application number: PCT/US2012/058595
Authority: WO
Inventors: Xiangmin Liao; Neil David Pearson; Israil Pendrak; Masayuki Sano
Original assignee: Glaxo Group Limited; Shionogi & Co., Ltd.
Priority date: 2011-10-04
Filing date: 2012-10-04
Publication date: 2013-04-11
Also published as: TW201329088A; RU2014117658A; CN104039152A; KR20140089369A; US9340556B2; AU2012318712B2; US20140249126A1; CA2851263A1; EP2763539A4; JP2014528965A; EP2763539A1; ES2623501T3; EP2763539B1; AU2012318712A1; BR112014008067A2; TWI547496B

Abstract

The present Invention relates to cephalosporin antibacterial compounds of Formula (!): corresponding pharmaceutically acceptable salts thereof, corresponding pharmaceutical compositions, compound preparation and treatment methods for bacterial infections, especially those caused by gram-negative bacteria.

Description

PU85023

ANTIBACTERIAL COMPOUNDS

FIELD OF TH E 1 ¥ΕΝΉΟΝ

The present Invention relates to cephalosporin antibacterial compounds, corresponding pharmaceutical compositions, compound preparation and treatment methods for bacterial infections.

BACKGROUND OF THE INVENTION

Over the past several decades, the frequency of antimicrobial resistance andts association with serious infectious diseases has increased at alarming rates.

For example, in the United States, the Centers for Disease Control and

Prevention estimate that roughly 1.7 million hospital-associated infections, from all types of microorganisms, including bacteria, combined, cause or contribute to 99,000 deaths each year. In Europe, where hospital surveys have been conducted, the category of Gram-negative infections are estimated to account for two-thirds of the 25,000 deaths each year. Nosocomial infections can cause severe pneumonia and infections of the urinary tract, bloodstream and other parts of the body. Many types are difficult to attack with antibiotics, and antibiotic resistance is spreading to Gram-negative bacteria that can Infect people outside the hospital (see, Pollack, Andrew. "Rising Threat of Infections Unfazed by Antibiotics" New York Times, Feb. 27, 2010). This high rate of resistance Increases the morbidity, mortality, and costs associated with nosocomial infections.

The problem of antibacterial resistance is compounded by the existence of bacterial strains resistant to multiple antibacterials. It is conventionally taught in the art that among:

Gram-positive organisms, resistant pathogens Include methicillin-(oxaciliin) resistant Staphylococcus aureus, beta-iactam-resistant and muitidrug- reslstant pneumococci, and vancomycin-resistant enterococci; and that Gram-negative resistance includes extended-spectrum beta-!actamases (ESBLs) in Klebsiella pneumoniae, Escherichia coii, and Proieus mirabilis, high-level third-generation cephalosporin (Amp C) beta-lactamase resistance among Enterobacter species and Citrobacter freundii, and muitidrug-resistance genes observed in Pseudomonas aeruginosa, Acinetobacter, and Stenotrophomonas maltophilia.

To date, a variety of beta-lactam drugs have been developed and beta-lactam drugs have become clinically extremely important antimicrobial drugs.

However, there are increasing number of bacteria! types which have obtained resistance against β-lactam drugs by producing β-lactamase, which degrade β-lactam drugs.

According to the Ambler molecular classification, β-lactamases are largely classified Into four classes. Specifically, these are Class A (TEM type, SHV type, CTX-M type and PU85023 the like), Class B (IMP type, VIM type, L-1 type and the like), Class C (AmpC type) and Class D (OXA type and the like). Amongst these, Classes A, C and D types are largely classified Into serines-lactamase, and on the other hand, Class B type is classified into meta!lo-f3-lactamase. It has been known that both have respectively different mechanisms to each other in terms of hydrolysis of β-lactam drugs.

Recently, clinical problems have been occurring due to the existence of Gram negative bacteria which have become highly resistant to β-iactam drugs including Cephems and Carbapenems, by production of Class A (ESBL) or D type serine-β- lactamases and Class B type metallo-p-lactamases which have extended their substrate spectrum. Particularly, metal!o- -iactamases are known to be one of the causes of obtaining multi-resistant Gram negative bacteria. Cephem compounds which exhibit Intermediate activity against metallo-p-lactamase producing Gram negative bacteria are known (i.e., e.g., see, international Patent Publication No. WO 2007/1 1951 1 to Asteiias Pharma. Inc. and Wakunaga Pharmaceuticals I nc. , Intern . '! Pub. Date: October 25, 2007 and Takeda et aL, "In Vitro Antibacterial Activity of a New Cephalosporin, FR 295389, against !MP-type etailo-p-Lactamase-Producters", The Journal of Antibiotics, vol. 81 , pp.38-39 (January 2008)).

However, there is a demand for development of Cephem compounds which exhibit more potent antimicrobial activity, in particular effectiveness against a variety of β- lactamase producing Gram negative bacteria.

One of the known antimicrobials to have poetnt anti-Gram negative bactericidal activity are Cephem compounds having a catechol group intramoiecuiarly (i.e., e.g., see Sanada et aL, "Comparison of Transport Pathways of Catechoi-Substituted

Cephalosporins, BO- 236 AND BO- 341 , Through the Outer Membrane of

ESCHERICHIA COL ", The Journal of Antibiotics, voi.43, No. 12, pp.1617-1620 ( 990); Weissberger et al, "L-658, 310, A New injectable Cephalosporin i. In Vitro Antibacterial Properties", The Journal of Antibiotics, vol.42, No. 5, pp.795-808 (1989); Okita et aL, "Synthesis and Antibacterial Activity of Cephalosporins Having a Catechol In the C3 Side Chain", The Journal of Antibiotics, vol.48, No. 5, pp.833-839 (1993)). The action thereof is that the catechol group forms a chelate with Fe ⁺, thereby the compound Is efficiently incorporated into the bacterial body by means of the Fe³⁺ transportation system across the cellular membrane (tonB-dependent iron transport system).

Thus there is a need for new antibacterials, particularly antibacteria!s with novel mechanisms of action. In light of the above, a need exists to develop compounds of the present

Invention, which provides Cephem compounds that exhibit potent antimicrobial spectrum against a variety of bacteria including Gram negative bacteria and/or Gram positive bacteria, corresponding pharmaceutical compositions and treatment methods for bacterial infections. More importantly, there is a need to develop Cephem compounds of the present invention, which exhibit:

o potent antimicrobial activity against beta-!actamase producing Gram negative bacteria;

o potent antimicrobial activity against multi-drug resistant microbes, in particular, Class B type meta!lo-beta-!actamase producing Gram negative bacteria; o effective antimicrobial activity against extended-spectrum beta-lactamase

(ESBL) producing bacteria; and

o a lack of cross-resistance against known Cephem drug or Carbapenem drugs.

The present Invention is directed to overcoming these and other problems encountered in the art.

SUMMARY OF THE SMVE TfOM

In general, the present invention relates to cephalosporin antibacterial compounds, corresponding pharmaceutical compositions, compound preparation and treatment methods for bacterial infections, especially those caused by gram-negative bacteria and gram-positive bacteria.

In particular, the present invention relates to novel compounds of Formulas (I) to (IX), respectively, or pharmaceutically acceptable salts thereof and corresponding pharmaceutical compositions, respectively.

The present invention also relates to processes for making compounds of

Formulas (I) to (IX), respectively, or pharmaceutically acceptable salt thereof.

The present invention also relates to methods for treating bacte ial infections, which comprises administering to a subject in need thereof an effective amount of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof or a corresponding pharmaceutical composition, respectively.

DETAILED DESCRIPTION! OF THE j VEMTjQM In general, the present invention relates to cephalosporin antibacterial compounds, corresponding pharmaceutical compositions, compound preparation and treatment methods for bacterial infections, especially those caused by gram-negative bacteria and qram-positive bacteria. PU85023

In particular, the present invention relates to novel compounds of Formulas (I) to (IX), respectively, or pharmaceutically acceptable salts thereof.

In one aspect, the present invention relates to a compound of Formula (I):

wherein:

W is -CH₂-, -S-, or -0-;

U is -CH₂-, -S-, or -O- ;

provided that:

when W is -CH₂~, U is -CH₂-, -S-, or -O- ;

when W is -S- or -0-, U is -CH₂-;

X Is N, or C-R^a;

R¹ and R² each are hydrogen, (C .₆)aikyl, or (CH₂)p-C(0)OR^b;

R³ and R⁴ each are hydrogen, OH or OR^c;

wherein:

R is hydrogen or halogen;

R^b or R^c each is H, (C-i„₆)-aikyi or an alkali metal;

R ⁹ Is hydrogen, -OCH₃, or -NH-NH(=0);

A Is R⁵ or -NR^dC(0)R⁵

wherein:

R^d is H or (C-e)-alkyl

R⁵ is an optionaiiy saturated or unsaturated monocyclic heterocyclic ring or an optionally saturated or unsaturated bi-cyc!ic or fused heterocyclic ring;

wherein:

each monocyclic heterocyclic ring has from 3 to 7 ring atoms and contains up to four heteroatoms;

each fused heterocyclic ring optionally includes carbocyclic rings or heterocyclic rings of up to four heteroatoms;

R^a optionally is substituted by one or more of the following substituenis PU85023 selected from -H, -OH, Oxo (=0), -CN, -N0₂,-halogen, - straight or branched Ci_₆ alkyl,- straight or branched Ci_₆ haloalkyi, C₃.₆-cycloalkyl, - straight or branched Ci. 6 straight or branched aikoxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -Q(CH₂)_yOR^g, - NR^hR' , -S0₂R^j, -S(CH_z)_qR^k, -NR'CiOJR"¹, aryl or heteroaryi;

wherein:

hetero atoms are selected from oxygen, nitrogen or sulphur;

carbocyclic rings or heterocyclic rings for each fused heterocyclic ring systems include non-aromatic rings or aromatic rings;

monocyclic heterocyclic rings or fused heterocyclic rings include substituted aromatic and non-aromatics;

each R^e, R^f, R⁹, R^h, R\ R , R^k, R^! ,or R^m each is selected from H, C-i alkyl, C ha!oa!kyl, d.₆ aikoxy;

each aryl or heteroaryi as defined above optionally is substituted H, -OH, -CN, -N0₂, -halogen, C _-6 -aiky!, C^ -haloalkyl, C^e -alkoxy, - OC(0)OH, -OC(0)Rⁿ, ~C(Q)OR^,:i, -0(CH₂)_y.QR^p, -NR^qR^r , -S0₂R^s, ~S(CH₂)_y~

R^s, -NR^uC(0)R^'";

wherein:

R", R°, R^p, R^q, R^r, R^s, R¹ ,R^U, or R^v each are selected from C-|.₆ aikyi, C-i„₆ ~haloalkyi, or Ci.₆~aikoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

In one aspect, the present invention relates to a compound of Formula (I), where W is -CH₂-. and U is -S- ; or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound of Formula (I), where W is ~-CH₂-, IS is -S-, and R ⁹ is hydrogen; or a pharmaceutically acceptable salt thereof.

In one aspect, the present invention relates to a compound of Formula (I), which definition referred to herein may include, but Is not limited to the following related sub-generic Formulas (I!) to (IX).

PU85023

In one aspect, the present invention relates to a compound of Formula (I I):

where:

X is N, or C-R^a;

R¹ and R² each are hydrogen , (C .₆)alkyl, or (CH₂)p-C(0)OR^b;

R³ and R⁴ each are hydrogen , OH or QR^C;

where:

R is hydrogen or halogen ;

R or R^c each is H , (Ci_6)-alkyl, an alkali metal or negative charge;

A Is R⁵ or -NR^dC(0)R⁵

where R° is H or (Ci_₆)-alky!

R⁵ is an optionaliy saturated or unsaturated monocyclic heterocyclic ring or an optionally saturated or unsaturated bl-cyc!ic or fused heterocyclic ring ;

where:

each fused heterocyclic ring optionally includes carbocyclic rings or heterocyclic rings of up to four heteroatoms; and

R^a optionally is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-haiogen , - straight or branched Ci_₆ aikyl, C₃„₆cyc!oa!kyi, - straight or branched C^e ha!oa!kyi, - straight or branched C-μ 6 straight or branched alkoxy, -OC(0)OH, -OC(Q)R^e, -C(0)OR^f, -Q(CH₂)_yOR⁹, - NR^hR' , -S0₂R^j, -S(CH₂)_qR^k, -NR C(0)R^";, aryl or heteroaryl;

where:

heteroatoms are selected from oxygen, nitrogen or sulphur;

monocyclic heterocyclic rings or fused heterocyclic rings include substituted aromatic and non-aromatics; each R^e, R^f, R⁹, R' R^!, R^j , R^K, R¹ ,or R^rn each is selected from H, Ci_6 alkyl, Ci.₆ ha!oa!kyl, Ci .₆ aikoxy;

each aryl or heteroaryl as defined above is optionally substituted with one or more of the following substituents selected from H , -OH , ~CN, N0₂,-halogen, C^ -alky!, C-i ₆ -haloa!kyl, C.,.₆ -alkoxy, -OC(0)OH , - OC(0)Rⁿ, -C(0)OR°, -0(CHz)y.OR^p, -NR^qR^r , -S0₂R^s, -S(CH2)y-R', - NR^uC(0)R^v;

where:

R", R°, R^p, R^q, R^r, R^s, R^} ,R^U, or R^v each are selected from C _-6 alkyl, C_1-6 -haioaikyi, or d_-s-alkoxy;

n, m. o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

in another aspect, the present invention relates to a compound of Formula (H), where X is C-R^a, where R^a is hydrogen or halogen.

in another aspect, the present invention relates to a compound of Formula (H), where X is C-R^a and R^a is hydrogen .

In one aspect, the present invention relates to a compound of Formula (I I I):

where:

X is N, or C-R^a;

R¹ and R² each are hydrogen , (Cvejaikyl, or (CH₂)p-C(0)OR^b:

R³ and R⁴ each are hydrogen , OH or OR^c;

where:

R^a is hydrogen or halogen ;

R^B or R^c each is H , (C-i-K)-a!kyi, an alkali metal or negative charge;

A is R⁵ or -NR^dC(0)R⁵

where R^d Is H or (C_1-6)-a!kyl

R⁵ is a monocyclic 3 to 7 membered heterocyclic ring or a bicyclic 10 membered heterocyclic ring ; PU85023 where:

each 3 to 7 membered heterocyclic ring contains up to four heteroatoms; each blcyclic 10 membered heterocyclic ring contains up to four heteroatoms;

where:

each heteroatom is selected from oyygen, nitrogen or sulphur; each 10 membered heterocyclic ring optionally contains carbocyclic rings or heterocyclic rings;

where:

carbocyclic rings or heterocyclic rings for each 10 membered heterocyclic ring systems contain non-aromatic rings or aromatic rings;

R⁵ optionally is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-haiogen, - straight or branched Ci_₆ alkyl,- straight or branched Ci_₆ aloalky!, C₃.₆-cycioaik l, - straight or branched C

₆ straight or branched aikoxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -0(CH₂)_yOR⁹, - NR^hR' , -S0₂R^j, -S(CH₂)_qR^k, -NR C(0)R ^'\ aryl or heteroaryi;

where:

each R^e, R^f, R⁹, R , R^!, R^j , R^k, R¹ ,or R^m as defined above is selected from H, Ci_₆ alkyl, C^e haloalky!, C^e a!koxy;

each aryl or heteroaryi as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - N0₂, -halogen, C^ -alky!, C^ -haloaikyi, C_1-6 -aikoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)_y.OR^p, -NR^qR^r , -S0₂R^s, -S(CH₂)_y-R^l, - NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each as defined above are selected from Ci.,₆ alkyl, C -haloalkyi, or Ci -alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof,

in another aspect, the present invention relates to a compound of Formula (III), where X is C-R^a, where R^a is hydrogen or halogen.

In another aspect, the present invention relates to a compound of Formula (ill), where X is C-R^a and R^a is hydrogen,

In another aspect, the present invention relates to a compound of Formulas (i) to PU85023

(IX) as defined herein , where A, may Include, but is not limited to:

where:

Re_> R₇, R₈, Rg, Rg', Rg ^;, Rg-, R10, R1 , R12 O!^" R 7 is H, straight or branched(C |.₆)- alkyi or G₃-6-cycioaikyl;

each A optionally further is substituted by one or more of the following

substituents selected from -H, -OH, Oxo (~0), -CN, -N0₂,-ha!ogen, - straight or branched C1-6 aikyl,- straight or branched d-e aioaikyi, Ca^-cycloalkyi , - straight or branched Ci_6 straight or branched alkoxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -0(CH₂)_y0R⁹, -NR^hR^; , - S0₂R^j, -S(CH₂)_qR^k, -NR'CiOJR"¹, aryi or heteroaryl;

where:

each R^e, R^f, R⁹, R^h, R^;, R^j , R^k, R¹ ,or R^m as defined above is selected from H, C-| .₆ aikyl, Ci.₆ haloalkyl, C- .₆ alkoxy;

each aryi or heteroaryl as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, -

NO_z, -halogen, C_1-e -aikyl, C -ha!oaikyl, Ci_₆ -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)_yX}R^p, -NR^qR^r , -S0₂R^s, -S(CH₂)_y-R^, _I - NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R" each as defined above are selected from Ci-e aikyl, Ci-e -haloalkyi, or Ci-₆~alkoxy; and PU85023 n, m, o, p, q or y each are 0 or an integer from 1 to 5.

In another aspect, the present invention relates to a compound of Formulas (I) to {IX), respectively, as defined herein, where A, may induce, hut is not limited to:

where:

i3, F¾7, F¾9,

Fin or -j₂, R 3> R _*, io, F¾-I 17, Ri8> Ri9; ₂0i 22! o R₂3 is H, straight or branched(Ci-6)-al yl or C₃-6-cyc!oa!kyl .

each A optionally further is substituted by one or more of the following

suhst!tuents selected from -H, -OH, Oxo (=0), -CN , -N0₂,-ha!ogen, - straight or branched Ci -6 aikyi,- straight or branched Cv.₆ a!oa!kyl, C₃.6~cycloalkyl , - straight or branched Ci.₆ straight or branched aikoxy, -OC(0)OH, -OC(0)R^s, -C(0)OR^f, -0(CH₂)_yOR⁹, -NR^"R , - 80₂R^j, -S(CH₂)_qR^k _! -NR'C(0)R^m, aryi or heteroaryl;

where:

each R^e, R^f, R⁹, R , R^!, R^j , R^k, R¹ ,or R^m as defined above is selected from H, Ci_₆ aikyl, C^e haloalky!, C^e a!koxy;

each aryi or heteroaryl as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - PU85023

N0₂, -halogen, C _-6 -aiky!, C^ -haloalkyi, C-i.e -alkoxy, -OC(0)OH, - OC(0)R^r', -C(0)OR°, -0(CH₂)y-OR^p, -NR^qR^r , -S0₂R^s, -S(CH₂)_y-R⁽, - NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^E, R* ,R^U, or R each as defined above are selected from Ci .₆ aikyl, d-e -haioaikyl, or Ci..e-alkoxy; and n, m, o, p, q or y each are 0 or an integer from 1 to 5. in another aspect, the present invention relates to a compound of Formula (I where:

Ra is hydrogen;

R¹ and R each are (C_1-S)alkyl;

R³ and R⁴ each are OH or OR^c;

where:

R^c is H, (Ci_₆)-alkyl or an alkali metal;

A Is

R or R is H, -straight or branched(Ci_₆)-alky! or -C₃.₆-cycloalkyl;

n is 1 ; or

a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound of Formula (IV):

where:

X is C-R^a;

R¹ and R² each are hydrogen, (Cvejaikyl, or (CH₂)p-C(0)OR^b

R³ and R⁴ each are hydrogen, OH or OR^c; PU85023 where:

R^a is hydrogen or halogen;

Rⁿ or R^c each is H, (Ci_₆)-alkyl, an alkali metal or negative charge;

A is R⁵ or -NR^dC(0)R⁵

where R^d is H or (C< _₆)-a!kyl

R⁵ is an optionally saturated or unsaturated monocyclic heterocyclic ring or an optionally saturated or unsaturated b!-cyeiic or fused heterocyclic ring;

where:

R⁵ optionally Is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-haiogen, - straight or branched Ci_₆ alkyi,- straight or branched C-^ haloalkyL C₃.₆-cycioaikyl, - straight or branched C

₆ straight or branched aikoxy, -OC(0)OH, -OC(0)R^e, ~C(0)OR^f, ~0(CH₂)_yOR⁹, - NR^hR' , -S0₂R^j, -S(CH₂)_qR^k, -NR C(0)R ^'\ aryl or heteroaryi;

where:

hetero atoms are selected from oxygen, nitrogen or sulphur;

each R^e, R^f, R⁹, R^h, R^!, R^j , R^k, R¹ ,or R^m each is selected from H, Ci-6 alkyl, Ci-₆ ha!oa!kyl, Ci-_B aikoxy;

each aryl or heteroaryi as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - N0₂,-haiogen, C _-6 -aikyl, d -e -haloalkyi, Ci_₆ -aikoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, ~0(CH_z)_y.OR^p, -NR^qR^r , -S0₂R^s,

- NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each are selected from Ci_6 alkyl, Ci.₆ -haloalkyi, or Ci_-6-alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof. PU85023

In another aspect, the present inveniion relates to a compound of Formula (IV), where X is C-R^a and R^s is hydrogen .

In another aspect, the present inveniion relates to a compound of Formula (V):

where:

X Is C~F ;

R¹ and R² each are hydrogen , (C₁.₆)alkyi, or (CH₂)p-C(0)OR^b;

R³ and R⁴ each are hydrogen , OH or OR^c;

where:

R³ is hydrogen or halogen ;

R^B or R^c each is H , (C-i_₆)-aikyi, an a!kail metal or negative charge;

A is R⁵ or ~NR^dC(0)R⁵

where R^d Is H or (C -e)-a\ky\

R⁵ is a monocyc!ic 3 to 7 membered heterocyclic ring or a bicyclic 10 membered heterocyclic ring ;

where:

each 3 to 7 membered heterocyclic ring contains up to four neteroatoms; each bicyclic 10 membered heterocyclic ring contains up to four heteroatoms;

where:

heteroatoms are selected from oyygen, nitrogen or sulphur; each 10 membered heterocyclic ring optionally contains carbocyciic rings or heterocyclic rings;

where:

carbocyciic rings or heterocyclic rings for each 10 membered heterocyclic ring systems contain non-aromatic rings or aromatic rings; PU85023

Rⁿ optionally is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-halogen, - straight or branched Ci_₆ alky!,- straight or branched Ci_₆ haloalkyi, C₃.₆-cycloalkyl,- straight or branched C-,. straight or branched alkoxy, -OC(0)OH, ~OC(0)R^e, -C(0)OR^f, -0(CH₂)_yOR⁹, - NR^hR' , -S0₂R^j, -S(CH_z)_qR^k, -NR'CiOJR"¹, aryl or heteroaryi;

where:

each R^e, R^f, R^g, R^h, R¹, R^j , R^K, R¹ ,or R^m as defined above is selected from H, C _-6 alkyl, C-i„₆ haloalkyL Ci_₆ alkoxy;

each aryl or heteroaryi as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, N0₂,-halogen, C_1-6 -alkyl, C^ -haloaikyi, C_1-s -alkoxy, -OC(0)OH, - OC(0)R'\ -C(0)OR°, -0(CH₂)_y„OR^p, -NR^qR^r , -S0₂R^s, -S(CH₂)_y-R^t, - NR^uC(0)R ;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each as defined above are selected from Ci_₆ alkyl, CT .₆ ~haloalkyi, or C-i.₆-aikoxy:

n, m, Q, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound of Formula (V), where X is C-R^a and R³ is hydrogen.

In another aspect, the present invention relates to a compound of Formula (VI):

where:

X Is C-R^a:

R¹ and R² each are hydrogen, (C<.₆)aikyi, or (CH₂)p-C(0)OR^b;

R³ and R⁴ each are hydrogen, OH or OR^c;

where:

R³ is hydrogen or halogen;

R^B or R^c each is H, (C _-6)-alkyl, an alkali metal or negative charge; PU85023

A

where:

Re> R₇, R₈, Rg, Rio. R-i i , Ri₂ or R₁₇ is H, straight or branched(Ci.₆)-alkyl or C₃.₆- cycloalkyi;

each A optionally further is substituted by one or more of the following

substituents selected from -H, -OH, Oxo (~0), -CN, -N0₂,-ha!ogen, - straight or branched C1 aikyl,- straight or branched d-e aioaikyi, Ca-s-cycloalkyi, - straight or branched Ci straight or branched alkoxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -0(CH₂)_y0R⁹, -NR^hR^; , - S0₂R^j, -S(CH₂)_qR^k, -NR'CiOJR"¹, aryi or heteroaryl;

where:

each R^e, R^f, R⁹, R^h, R^;, R^j , R^k, R¹ ,or R^m as defined above is selected from H, C-| .₆ aikyl, Ci.₆ haloalkyl, C alkoxy;

NO_z, -halogen, C_{1 -e} -aikyl, C -ha!oaikyl, Ci_₆ -alkoxy, -OC(0)OH , - OC(0)R^R, -C(0)OR°, -0(CH_zVOR^p, -NR^qR^r , -S0₂R^s, -S(CH₂)_y-R^, _I - NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R¹ ,R^U, or R^v each as defined above are selected from Ci-₆ aikyl, Ci.₆ -haloalkyl, or Ci._B~aikoxy: and PU85023 n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt: thereof.

In another aspect, the present invention relates to a compound of Formula (VII):

where:

X Is C-R⁸;

R¹ and R² each are hydrogen, (Chalky!, or (CH₂)_p-C(0)OR^b;

R³ and R^A each are hydrogen, OH or OR^c;

where:

R³ is hydrogen or halogen;

R^B or R^c each is H, (Ci_-6)-aikyl, an alkali metal or negative charge;

- 18 - PU85023 where:

R g, Py, Qg, P Kg, R Kio, O r<ii n ΟrΓ Ώ rV|2,

P K23

Is H, straight or branched(C>..₆)-alkyl or C₃..₆-cydoa!kyl.

each A optionaiiy further is substituted by one or more of the following

substituents selected from -H, ~OH, Oxo (=0), ~CN, -N0₂,-halogen, - straight or branched C'i-6 aikyl,- straight or branched C-j-e haioaikyl, C₃.₆-cyc!oa!kyi, - straight or branched C-|..₆ straight or branched alkoxy, -OC(0)OH, -OC(Q)R^e, -C(0)OR^f, -Q(CH₂)_yOR⁹, -NR R , - SO;.: -S(CH₂)_qR^k, -NR'C(0)R^m, aryi or heteroaryl;

where:

each R^e, R^f, R⁹, R^h, R\ & , R^k, R^! ,or R^m as defined above is selected from H, C_1-6 alkyl, C _-6 aloalkyl. Ci_₆ alkoxy;

each aryl or heteroaryl as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - NOz.-halogen, C,^ -alkyl, C^ -haloalkyl, C_1-6 -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)_y..OR^p, -NR^qR^r , -S0₂R^s, -SiCH₂)_y~R\ - NR^uC(0)R^v;

where:

R", R°, R^p, R^q, R^r, R^s, R¹ ,R^U, or R^v each as defined above are selected from C _-6 alkyl, Ci_₆ -haioaikyl, or C _-6-alkoxy; and n, n, o, p, q or y each are 0 or an integer from 1 to 5,

In another aspect, the present invention relates to a compound of Formula (VII) where:

Ra is hydrogen;

R and R² each are (C_1-6)alkyl;

R³ and R⁴ each are OH or OR⁰;

where:

R^c is H, (C"].fi)-aikyl, an alkali metal or negative charge;

A Is

R⁸ or R ^'2 is H, (Ci..₆)-alkyl, or C₃.₆-cycloaiky!;

n is i ; or PU85023 a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound of Formula (VI I I):

where:

R and R² each are hydrogen , (Ci_₆)aikyl, or (CH₂)p-C(0)OR^b;

R^J and R⁴ each are hydrogen , OH or OR^c;

where:

R or R^c each is H , (C _-6)-alkyl, an alkali metal or negative charge;

A Is R⁵ or -NR^dC(0)R⁵

where R^a is H or (Ci_₆)-alkyl

R⁵ is an optionaliy saturated or unsaturated monocyclic heterocyclic ring or an optionaliy saturated or unsaturated bl-cyciic or fused heterocyclic ring ;

where:

each fused heterocyclic ring optionally includes carbocyciic rings or heterocyclic rings of up to four heteroatoms;

R5 optionally is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-haiogen , - straight or branched C _-6 a!kyl,- straight or branched C_1-6 haloalkyL C:3__B-eycioaiky! ,- straight or branched C,.₆ straight or branched alkoxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, ~0(CH₂)_yQR⁹, - NR^hR' , -S0₂R^j, -S(CH₂)_qR^k, -NR^!C(Q)R^rn, aryl or heteroaryl;

where:

hetero atoms are selected from oxygen , nitrogen or sulphur;

carbocyciic rings or heterocyclic rings for each fused heterocyclic ring systems include non-aromatic rings or aromatic rings;

each R^e, R^f, R⁹, R , R^!, R^j , R^k, R¹ ,or R^m each is selected from H, PU85023

Ci_6 alkyl, Ci.₆ ha!oa!kyl, Ci .₆ aikoxy;

each ary! or heteroary! as defined above optionally Is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, -NO2, -halogen, C^e -alkyl, Ci_-6 -ha!oaikyl, C^ -alkoxy, - OC(0)OH, -OC(0)Rⁿ, -C(0)OR°, -0(CH_z)_y.OR^p, -NR^qR^r , -S0₂R^s, -S(CH_z)_y- R\ -NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each are selected from C1-6 alkyl, C-i -haloalky!, or Ci_6-alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

in another aspect, the present invention relates to a compound of Formula (VIII), where X is C-R^a and R^a is hydrogen.

in another aspect, the present invention relates to a compound of Formula (VIM),

In another aspect, the present invention relates to a compound of Formula (IX):

where:

R¹ and R² each are hydrogen, (C_1-6)alkyl, or (CH₂)p~C(0)QR^b:

R³ and R⁴ each are hydrogen, OH or OR^c;

wnere:

R or R^c each is H, (Ci_6)-alkyl, an alkali metal or negative charge: PU85023

A is R⁵ or -NR^dC(0)R⁵

where R^a is H or (Ci_₆)-alk l

R⁵ is a monocyclic 3 to 7 membered heterocyclic ring or a bicyciic 10 membered

heterocyclic ring;

where:

each 3 to 7 membered heterocyclic ring contains up to four heteroatoms; each bicyciic 10 membered heterocyclic ring contains up to four heteroatoms; where:

heteroatoms are selected from oyygen, nitrogen or sulphur;

each 0 membered heterocyclic ring optionally contains carbocyc!ic rings or heterocyclic rings;

where:

carbocyciic rings or heterocyclic rings for each 0

membered heterocyclic ring systems contain non-aromatic rings or aromatic rings;

R⁵ optionaiiy Is substituted by one or more of the foilowing substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-halogen, - straight or branched Ci.₆ aikyl,- straight or branched Ci-₆ haloalkyl, C₃.6-cycloalkyl, - straight or branched Ci . 6 straight or branched aikoxy, -OC(0)OH, -OC(0)R^e, -C(Q)OR^f, -0(CH₂)_yQR^s, - NR^hR' , -S0₂R^j, -S(CH₂)_qR^k, -NR'C(0)R^m, aryl or heteroaryi;

where:

each R^e, R^f, R⁹, R^h, R^!, R^j , R^k, R^l ,or R^m as defined above is selected from H, C _-6 aikyl, C_1-6 aloalkyi, C,„6 aikoxy;

each aryl or heteroaryi as defined above is optionaiiy substituted with one or more of the following substituents selected from H, -OH, -CN, -

N0₂, -halogen, C _-6 -aiky!, C^ -haloalkyi, C^e -alkoxy, -OC(0)OH, - OC(0)R^r', -C(0)OR°, -0(CH₂)y-OR^p, -NR^qR^r , -S0₂R^s, -S(CH₂)_y-R⁽, - NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^E, R* ,R^U, or R each as defined above are selected from Ci .₆ aikyl, d-e -haloalkyl, or Ci..e-alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound of Formula (IX), where X is G-R^a and R^a is hydrogen. PU85023

In another aspect, the present invention relates to a compound of Formula (IX), where

where R⁸ or R¹² is H, (Ci_₆)-a!kyi or C₃.₆-cycloaikyl.

In another aspect, the present invention relates to a compound or compound species which is :

1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy

dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetami

(((4-methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2 ]oct-2-en-3-yl)methyi)-1 - (2-(5-chloro-6 -bis((4-methoxybenzyi)oxy)-1 -oxo-3,4-di ydroisoquinolin-2(1 H)- y!)ethyi)pyrrolidin-1 -ium;

(6R7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-(2-(5-hydroxy-1-methyl-4-oxo-1 ,4-dihydropyridine-2-carboxamido) ethyl)pyrrolidin-1-ium-1-yl)methy!)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)~7-((Z)~2-(2~aminothiazol~4-yi)~2-(((S)-1 ,2~dicarboxyethoxy)imino)

acetamido)-3-((1 -(2-(7~hydroxy-1 ,8-dioxo-3,4-dihydi -1 H~pyrido[1 ,2-a]pyrazin-2(8H)- yi)ethyi)pyrrolidin-1 -ium-1-yi)methyi)-8-oxo-5-thia-1-azabicycio[4,2.0]oct-2-ene-2- carboxylate;

(6R R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino)

acetamido)-3-((1 -(2-(7-hydroxy-1 _>8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- yi)ethyl)pyrrolidin-1 -ium-1-yl)methyi)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2- carboxylate:

(6R R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1 -((3-hydroxy-1 -methyi-4-oxo-1 ,4-di ydroquinolin-6-yi)methyl)pyrroii ium-1 -yi)methy!)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2-carboxyiate;

1 -(((6R JR)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-y!)oxy)imirio) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yi)methyl)-1-(2-(3-hydroxy- 1-methyi-4-oxo-1 ,4~dihydroquinoline~8-carboxanido)ethyi)pyrrolidin-1~!um;

1 -(((6R R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropar!-2-y!)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2,03oct-2-en-3-yi)methyl)-1-(2-(3- PU85023 hydroxy-1 -met yi-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)et yi)pyrrolidin-1-ium;

1 -(((6RJR)-7-((E)-3-(2-aminothiazol-5-y^

oxopropyi)-2-carboxy-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-en-3-yi)methyl)-1-((1-et yl- 8,7-dih droxy-4~oxo-1 ,4-dihydrocinnolin-3-yi)met yl)pyrroiidin-1-ium;

1 -(((6R_!7R)-7-((Z)-2-(2-aminothiazo!-4-yl)-2-(((R)-1 ,2-dicarboxyethoxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yi)methyi)-1-((1-ethyi- 6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi)pyrrol!din-1-ium;

1 -(((6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-y!)oxy)imino) acetamido)-2-carbaxy-8-oxo-5 hia-1-azabicyd^

6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)pyrrolidin-1-ium;

1 -(((6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yi)rnethyl)-1-(2-(1 -ethyl- 6 -dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxarriido)ethyi)pyrrolidin-1 -iLim;

1 -(((6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yi)methyl)-1-(2-(1 -ethyl- 6,7-dihydroxy-4-oxo-1 ,4-dihydroquino!ine-3-carboxamido)ethyl)pyrro!idin-1 -ium;

1 -(((6R,7R)-7-((E)-3-(2-aminothiazol-5-yl)-3-(((R)-1 ,2-dicarboxyethoxy) imino)-2- oxopropyi)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-eri-3-yi)methyl)-1-((1-ethy!- 6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)piperidin-1 -ium;

1 -(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)-

1-(2-(3-hydroxy-1 -methyl-4-oxo-1 ,4-dihydroquinoline-8-carboxamido)ethyl)pyrrolid!n-1-

!um;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((1-ethyl-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate:

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate; PU85023

(6R R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6 -dihydroxy^-oxo-l ^-dihydroquinolin-S- yQmethylJpyrroiidin-l-ium-l-y met y -e-oxo-S-thia-l-azabicyclo^^.Ojoct^-ene^- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((5-c loro-1 -ethyl-6,7-di ydroxy-4-oxo-1 ,4-di ydroquinolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxyiatopropan-2-yl)oxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-d!hydroxy-4-oxo-1 ,4-d!hydroqu!nolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazo!-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-arnino-5-chlorothiazo!-4-yl)-2-(((2-carboxypropari-2-y!)oxy) imino)acetamido)-3-((1 -((5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((1-ethyi-6 ,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyl)pyrrolidin- 1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((2-carboxypropan-2- yi)oxy)imino)acetamido)-3-((1-((1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxy!ate:

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetamido)-3-((1-((1 -et yl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R_>7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((S)-1 _>2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-ethyi-6 ,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyl)pyrrolidin- 1-ium-1-yi)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)im!no) PU85023 acetamido)-3-((1-(2-(5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyi)pyrroiidin-1-ium-1-yi)rriethyl)-8-oxo-5-t ia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetarnido)-3-((1-((1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoliri-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6RJR)-7-((Z)-2~(2-amlno-5-chioroihlazo!-4~yi)-2~(((S)-1 _>2-dicarboxyeihoxy) imino) acetamido)-3-((1 -((1-ethyi-6.7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi) methyi)pyrroiidin- 1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino} acetamido)-3-((1-(2-(1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2-carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-(2-(5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoiine-3- carboxamido)et yl)pyrroiidin-1-ium-2-ylium-1 -yl)methyi)-8-oxo-5-t ia-1-azabicyclo

[4,2.0 joci-2~en e-2~ca rboxy i aie ;

1 -(((6R,7R)-7-((Z)-2-(2-amino-5-chiorot iazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yl)methyi)-1-(2-(5- chioro-1-ethyi-6 J-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido)ethan-1-ylium-1- yl)pyrroiidin-1 -ium;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1.2-dicarboxyethoxy) imino)acetamido)-3-((1-(2-(5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3- carboxamido)ethyi)pyrroiidin-1-ium-2-yiium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo

[4.2.0]oct-2-ene-2-carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino) acetamido)-3-((1-((5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R_>7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((S)-1 _>2-dicarboxyethoxy) imino)acetamido)-3-((1-((5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4~dihydrocinno!in-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 _>2-dicarboxyethoxy) PU85023 imino)acetamido)-3-((1-((1-ethyl-5-fluoro-6,7-dihyclroxy-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)met yi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxy!ate:

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-((1-ethyi-5-f!uoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi) pyrrol idin-1-ium-1-yi)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate;

(6RJR)-7-((Z)-2~(2-aminothiazoi-4-y!)-2~(((S)-1 ,2-dicarboxyethoxy)imirio) acetamido)-3-((1-((5-fiuouro-1 -ethyi-6_>7-dihydroxy-4-oxo-1 _>4-dihydroquinolin-3-yi) methyl)pyrrolidin-1 -ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino} acetamido)-3-((1 -(2-(1-ethyl-5-fiuoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrroiidin-1-!um-1-yl)rnethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2-carboxyiate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1-((1-ethyl-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoliri-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-((1-ethyi-6-fiuoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi) pyrrol idin-1-ium-1-y!)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2,03oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-ethyl-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl) methyl) pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1 -(2~(1-ethy!-6 !uoro-7,8-dlhydroxy-4-oxo-1 ,4-dihydroquino!ine-3- carboxamldo)eihy!)pyrrolidin-1-ium-1-y!)methyi)-8-oxo-5-ihia-1~azabicyclo[4.2 3oct-2- ene~2-carboxy!ate;

(6R,7R)-7-((Z)-2-(2-amiriothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)!mir!o) acetamido)-3-((1-((1-ethyi-7,8-dihydroxy-4-oxo-1 ,4-di ydroquinoiin-3-yl)methyl)pyrroiy 1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2 -carboxylate;

(6R_>7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-(2-(1-ethyl-7,8-di ydroxy-4-oxo-1 ,4-di ydroquinoline-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2,03oct-2- ene-2-carboxyiate; PU85023

(6R7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-(2-(1-ethyl-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido) ethyl)pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-y!)-2-(((S)-1 ,2-dicarboxyethoxy)

imino)acetamido)-3-((1-(2-(1 -ethyl-5,6-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1 -azabicycio[4.2,03oct-2- ene-2-carboxyiate;

(6R₎7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-(tert-bLityl)-6,7-d!hydroxy-4-oxo-1 ,4-d!hydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiaie;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)

imino)acetamido)-3-((1-(2-(1-(tert-butyi)-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2~carboxy!aie;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-cyciopropyi-6,7-dihydroxy-4-oxo-1 ,4-di ydroquinoiin-3-yi) methyl) pyrrol idin-1-ium-1-y!)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2,03oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazo!-4-yl)-2-(((S)-1 _!2-dicarboxyethoxy) imino)acetamido)-3-((1 -((1 -cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-y!) methyl)pyrrolidin-1 -ium-1 -yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-((1 -((1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate:

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1 -(2-(1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate;

(6R_>7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imlno) acetamldo)-3-((1-((5-chloro-1 -cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate; PU85023

(6R7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((5-chloro-1 -cyclopropyl-67-dihydroxy-4-oxo-1 ,4-di ydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)met yi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1 -(2-(5-chloro-1 -cyclopropyi-6J-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2,03oct-2- ene-2-carboxyiate;

(6R R)-7-((Z)-2-(2-arninothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6 ,7-dihydroxy-1-methyi-4-oxo-1 ,4-dihydroquinolin-3-yl)methyl) pyrrol idin-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate;

(6R R)-7-((Z)-2-(2-arninothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6,7-dihydroxy-1-isopropyl-4-oxo-1 ,4-dihydroquinolin-3-yi)rnethyl) pyrrol idin-1-ium-1-yi)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2 3oct-2-erie-2-carboxylate;

1-(((6RJR)-7-((Z)-2-(2-amino-5-chlorot^

imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]od-2-en-3-yl)methyi)-1 -(2 chioro-1-ethyl-6 -dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido)ethyl)pyrroNdin-1 - ium;

1-(((6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]od-2-en-3-yl)methyl)-1 -(2-(1 ^ ethyl-8-fiuoro-6 -dihydroxy^-oxo-l ^-dihydroquinoline-S-carboxamidoJethylJpyrrolidin-l- lum;

1-(((6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicy^

ethyl-8-fluoro-6 -dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yi)methyl)pyrroiidin-1 -ium;

(6R R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-6,7-dihydroxy-1-methyl-4-oxo-1 ,4-dihydroquinoiin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-triia-1-azabicyclo[4.2.0]oGt-2-ene-2- carboxylate; or

a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound or compound species or a pharmaceutically acceptable salt, where the pharmaceuticaiiy acceptable salt is a sodium salt, a di-sodium salt or a trifluoroacetic acid salt.

In another aspect, the present invention relates to a compound or compound species which is : PU85023

1-(((6R,7R)-7-((Z)-2-((((S)- -(tert-butDxy)-1-((4-methoxybenzyl)oxy)-1 ,4- diaxobutan-2-yi)oxy)imina)-2-(2-((tert-butoxycar^^

(((4-methoxybenzyi)oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yl)methyi)-1 - (2-(5-chloro-6_!7-bis((4-methoxybenzyi)oxy)-1 -oxo-3,4-dihydroisoquino!in-2(1 H)- y!)ethyl)pyrrolidin-1 -ium, 2 sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamido)-3-((1 -(2-(5- ydroxy-1-methyl-4-oxo-1 ,4-di ydropyridine-2- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2,03oct-2- ene-2-carboxyiate, 3 Sodium salt;

(6R₎7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetam!do)-3-((1 -(2-(7-hydroxy-1.8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- yl)ethyl)pyrrolidin-1 -ium-1-yi)methyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1 -(2-(7-hydiOxy-1 _!8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- yl)ethyi)pyrrolidin-1 -ium-1-yi)met yl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2- carboxylate 3 Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminot iazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1 -((3-hydroxy-1 -methyi-4-oxo-1 _>4-dihydroquinolin-6-yi)methyl)pyrroiidin-1- ium-1 -yi)methyi)-8-oxo-5-thia-1-azabicycio[4,2.0]oct-2-ene-2-carboxyiate, 3 Sodium sait;

1 -(((6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yi)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2-en-3-yi)methyl)-1-(2-(3-hydroxy- 1-methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)ethyi)pyrrolidin-1-ium;

1-(((6R.7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2- yi)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)- 1-(2-(3-hydroxy-1 -methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)ethyi)pyrrolidin-1- ium, sodium salt;

1 -(((6R_!7R)-7-((E)-3-(2-aminothiazol-5-yi)-3-(((R)-1 _!2-dicarboxyethoxy)imino)-2- oxopropyi)-2-carboxy-8-oxo-5-thia-1 -azabicyc!o[4.2.03oct-2-en-3-yi)rnethyl)-1-((1-ethyl- 6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)met yl)pyrrolidin-1-ium, disodium sail;

1 -(((6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((R)-1 ,2-dicarboxyethoxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yi)methyi)-1-((1-ethyi- 6,7-dihydroxy-4-oxo-1 ,4-dihyd!OquiRoiiri-3~yi)methy!)pyn ildin-1-ium disodium salt;

1-(((6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)- PU85023

1-((1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)pyrrolidiri-1 -ium;

1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yi)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)- 1-(2-(1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido)ethyl)pyrrolidin-1 - ium;

1-(((6Κ,7Ρ)-7-((Ζ)-2-(2-3!Π!ηο1ήί3ζοί-4^Ι)-2-(((2-03^οχνρΓορ3η-2- y!)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)-1- (2-(1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido)ethyl)pyrrolidin-1 -ium;

1 -(((6R,7R)-7-((E)-3-(2-aminothiazol-5-yi)-3-(((R)-1 _>2-dicarboxyethoxy) imino)-2- oxopropyi)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-en-3-yi)methyl)-1-((1-ethyl- 6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)piperidin-1 -ium, disodium salt;

1 -(((6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yi)methyl)-1-(2-(3- hydroxy-1 -methyi-4-oxo-1 ,4-dihydroquinoline-8-carboxamido)ethyi)pyrrolidin-1-ium;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((1-ethyi-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-di ydroquinoiin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- earboxy!ate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, 2 Sodium sail;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxyiatopropan-2-yl)oxy)imino) PU85023 acetamido)-3-((1-((5-chloro-1 -ethyl-6 -dihydroxy^-oxo-l ^-dihydroquinolin-S- yQmethylJpyrroiidin-l-ium-l-y met y -e-oxo-S-thia-l-azabicyclo^^.Ojoct^-ene^- carboxy!ate, Sodium sail;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorot iazol-4-yl)-2-(((S)-1 _!2-dicarboxyet oxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-((1-((5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 _>4-dihydroquinolin- 3-yl)methyi)pyrrolidin-1-ium-1 -yi)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2- carboxyiaie, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((1-et yi-6 J~dihydroxy-4-oxo-1 ,4-dihydroqulnolin-3-yl)meihyl)pyrrolldln- 1-ium-1-yi)methyl)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazo!-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-((1-((1 -ethy!-6,7-di ydroxy-4-oxo-1 ,4-dihydroquino!in-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazo!-4-yl)-2-(((2-carboxypropan-2- yi)oxy)imino)acetamido)-3-((1-((1 -ethy!-6,7-dihydroxy-4-oxo-1 ,4-dihydroquino!in-3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate. Sodium salt;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1.2-dicarboxyethoxy) imino)acetamido)-3-((1-((1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-d!hydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)met yi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-(2-(5-chloro-1-ethyl-6_!7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- Garboxamido)ethy!)pyrroiidin-1-ium-1-y!)rriethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene~2-carboxylafe, sodium salt;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetamido)-3-((1-((1 -ethyi-6,7-di ydroxy-4-oxo-1 ,4-di ydrocinnolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino) PU85023 acetamido)-3-((1 -((1-ethyi-6,7-di ydroxy-4-oxo-1 ,4-di ydrocinnolin-3-yi) met yi)pyrroiidin- 1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)

imino)acetamido)-3-((1-(2-(1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3- Garboxamido)ethy!)pyrroi!din-1-ium-1-y!)rriethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2- ene-2-carboxylate, sodium salt;

(6RJR)-7-((Z)-2~(2-aminothiazoi-4-yl)-2-(((2~carboxypropan-2~yi)oxy) imino)acetamido)-3-((1 -(2-(5-chioro-1-ethyi-6J-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3- carboxamido)ethyl)pyrroiidin-1-ium-2-yliurn-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo

[4.2.0]oct-2-ene-2-carboxylate, Sodium salt;

1 -(((6R,7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)- 1-(2-(5-chioro-1-ethyi-6,7-di ydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido)ethan-1- ylium-1-yl)pyrro!idin-1-ium, Sodium salt;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1 -(2-(5-ch!oro-1-ethy!-6,7-di ydroxy-4-oxo-1 ,4-dihydrocinnoline-3- carboxamido)et yl)pyrroiidin-1-ium-2-ylium-1 -yl)methyi)-8-oxo-5-t ia-1-azabicyclo

[4,2.0 joci-2~en e-2~ca rboxy i aie ;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)

imino)acetamido)-3-((1 -((5-chloro-1-ethyi-6_>7-dihydroxy-4-oxo-1 ,4-dihydrocinno!in~3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate. Sodium sait;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1.2-dicarboxyethoxy) imino)acetamido)-3-((1 -((5-chloro-1-ethy!-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinno!in-3- yi)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxy!ate, Sodium sait;

(6R 7R)-7-((Z)-2~(2-am.no-5^

imino)acetamido)-3-((1-((1-ethyl-5-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoliri-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, Sodium sait;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-((1-ethyi-5-f!uoro-6_l7-dihydroxy-4-oxo-1 _l4-dihydroquinoiin-3-yl)methyi) pyrrol idin-1-ium-1-yi)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamidoJ-S-iil-iiS-fiuouro-l -ethyi-G -dihydroxy^-oxo-l ^-dihydroquinolin-S-yl) PU85023 methyl)pyrrolidin-1 -ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- earboxylate, Sodium sail;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)

imino)acetamido)-3-((1-(2-(1 -ethyi-5-fiuoro-6 J-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- Garboxamido)ethy!)pyrroiidin-1-ium-1-y!)rriethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2-carboxyiate, sodium salt;

(6R_>7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((S)-1 _>2-dicarboxyethoxy) imino)acetamido)-3-((1-((1-et yl-6-fluoro-7,8-di ydroxy-4-oxo-1 ,4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiaie, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamidoJ-S-iil-iil-ethyi-B-fluoro^^-dihydroxy^-oxo-l ^-dihydroquinoiin-S-y methyi) pyrrol idin-1-ium-1-yi)!Tiethyl)-8-oxo-5-t ia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate;

(6R R)-7-((Z)-2-(2-arriiriothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-et yi-6-fiuoro-7,8-diriydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl) methyl )pyrro!idin-1 -ium-1 -yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate, sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-3-((1-(2-(1 -ethyl-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyc!o[4.2.03oct-2- ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-ethyl-7,8-dihydroxy-4-oxo-1 ^-dihydroquinolin-3-yl)methyl)pyrrolidin- 1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate. sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-3-((1-(2-(1 -ethyl-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrrolidin-1-ium-1-yl)rriethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2- ene~2-carboxylate;

(6R,7R)-7-((Z)-2-(2-amlnothiazoi-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetamido)-3-((1-(2-(1-ethyl-7,8-dihydroxy-4-oxo-1 _!4-dihydroquinolirie-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2,03oct-2- ene-2-carboxyiate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 _l2-dicarboxyethoxy)

imino)acetamido)-3-((1-(2-(1 -ethyl-5,6-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethyl)pyrroiidin-1-!um-1-yl)methyl)-8-oxo-5-thia-1 -azab!cyclo[4.2.0]oct-2- PU85023 ene-2-carboxy!ate, 2 Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-(tert-butyi)-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 _l2-dicarboxyethoxy) imino) acetamido)-3-((1-(2-(1-(tert-butyl)-6_>7-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2,03oct-2- ene-2-carboxyiate, Sodium salt;

(6R₎7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-cyciopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yi) methyl) pyrrol !din-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate, Sodium salt;

(6R R)-7-((Z)-2-(2-amlno-5-chioroihlazo!-4-yi)-2-(((S)-1 ,2-dicarboxyeihoxy) imino)acetamido)-3-((1 -((1 -cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yi) methyl)pyrro!idin-1 -ium-1 -yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazo!-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imlno)acetamido)-3~((1 ~((1-cyclopi pyi~6,7-dihydroxy-4~oxo~1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1 -ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1 -(2-(1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate, 2 Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, 2 Sodium salt;

(6R7R)-7-((Z)-2-(2-amiriothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((5-c loro-1 -cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate, Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-(2-(5-chloro-1 -cyclopropyi-6J-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- PU85023 carboxamido)ethyi)pyrroiidin-1-ium-1-yi)methyl)-8-oxo-5-t ia-1-azabicyclo[4.2 3oct-2- ene-2-carboxyiate, Sodium salt:

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6,7-dihydroxy-1-methyi-4-oxo-1 _!4-dihydroquinoliri-3-yl)methyl) pyrrolidin-l-ium-l-ylJmethylJ-S-oxo-S-thia-l-azabicyclo^^.Ojoct^-ene^-carboxylate, 2 Sodium salt;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6,7-dihydroxy-1-isopropy!-4-oxo-1 ,4-dihydroquinolin-3-y!)methyl) pyrrol!din-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate, 2 Sodium salt;

1-(((6R_l7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1 -(2- (5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethyl)pyrrolidin-1-ium, Trifiuoroacetic acid salt;

1-(((6RJR)-7-((Z)-2-(2-amino-5-chlorothiazoM-yl)-2-(((SV1 ,2-dicarboxyethoxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]od-2-en-3-yl)methyi)-1-(2-(1- ethyl-e-fiuoro-BJ-dihydroxy^-oxo-l ^-dihydroquinoline-S-carboxamidoJethyiJpyrroiidin-l- iurn, Trifiuoroacetic acid salt;

1-(((6R,7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]od-2-en-3-yl)methyi)-1-((1 - ethyi-8-fluoro-6 -dihydroxy^-oxo-l ^-dihydroquinolin-S-yiJmethy pyrroiidin-l-ium, Trifiuoroacetic acid salt; or

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-6,7-dihydroxy-1-methyl-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- earboxy!ate, 2 Sodium salt;

In another aspect the present invention relates to a compound which is 1- (((6R R)-7-((Z)-2-(2-aminothiazol-4-yl)-2^

carboxy-8-oxo-5 hia-1-azabicyclo[4.2.0joct-2-en-3-yl)methyl)~1-((1-ethyl-6,7-dlhydroxy-4- oxo-1 ,4-dihydroquinolin-3-yl)methy!)pyrrolidin-1-ium disodium salt:

a pharmaceutically acceptable salt thereof ,

In another aspect, the present invention relates to a compound which is 1- (((6R7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)- 2-carboxy-8-oxo-5-thia-1~azabicycio[4.2 ]oci-2-en~3-yl)methyi)~1-((1-ethyi-6 ,7-dihydroxy- 4-oxo-1 ,4-dihydrocinno!in-3~yi)methyl)pyn ildin-1 -ium:

a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound which is 1- (((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)- 2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yl)methyi)-1-(2-(1-ethyi-6,7- dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido)ethyi)pyrrolidin-1-ium:

a pharmaceutically acceptable salt thereof, It is recognized that the compounds of Formulas (I) to (IX), respectively, or pharmaceutically acceptable salts thereof of the present invention as defined above may exist in forms as stereoisomers, regioisomers, or diastereiomers. These compounds may contain one or more asymmetric carbon atoms and may exist in racemic and optically active forms. For example, compounds of the present invention PU85023 may exist as a racemic mixture of R(+) and S(-) enantiomers, or in separate respectively optica! forms, i.e., existing separately as either the R(+) enantiomer form or in the S(+) enantiomer form. Ail of these individual compounds, isomers, and mixtures thereof are included within the scope of the present Invention.

SUBSTTTUENT OEFiMITiOISSS

As used herein, the term alkali metal is intended to mean the Group I elements, which include, but are not limited to lithium (Li), sodium (Na), or potassium (K) and the like. The term alkali earth metal include, but are not limited to calcium (Ca) or magnesium (Mg) and the like.

As used herein, the term "alky!" represents a saturated, straight or branched hydrocarbon moiety, which may be unsubstituted or substituted by one, or more of the subst!tuents defined herein. Exemplary alky!s include, but are not limited to methyl (Me), ethy! (Et), propyl, isopropy!, buty!, isobutyl, t-butyi, pentyl and the like. The term "C1 -C8" refers to an alky! containing from 1 to 8 carbon atoms.

When the term "alky!" is used in combination with other substituent groups, such as "haloalkyi" or "hydroxyalkyl", "arylalkyl", the term "alky!" is intended to encompass a divalent straight or branched-chain hydrocarbon radical.

The terms "halogen" and "halo" represent chloro, fiuoro, bromo or iodo substituents.

"Hydroxy" or "hydroxyl" is intended to mean the radical -OH.

For example, haloalkyi is intended to mean a saturated, straight or branched hydrocarbon moiety substituted with one or more halogen groups, where halogen is fluoro, chloro, bromo or iodo. Representative haloalkvis include, but are not limited to trifiuoromethyl (-CF₃). tetrafluoroethyi (-CF₂CHF₂), pentafluoroethyl (-CF₂CF₃) and the like. For example, hydroxyalkyl Is intended to mean a saturated, straight or branched hydrocarbon moiety substituted with one or more hydroxy groups. .

As used herein, the term "a!kenyl" refers to a straight or branched hydrocarbon moiety containing at least 1 and up to 3 carbon-carbon double bonds. Examples include ethenyl and propeny!.

As used herein, the term "aikynyi" refers to a straight or branched hydrocarbon moiety containing at least 1 and up to 3 carbon-carbon triple bonds. Examples include ethynyi and propynyl.

As used herein, the term "cycloalky!" refers to a non-aromatic, saturated, cyclic hydrocarbon ring. The term "(C₃-C₈)cycioaikyi" refers to a non-aromatic cyclic hydrocarbon ring having from three to eight ring carbon atoms. Exemplary PU85023

"(C3-C8)cycloalkyi" groups useful in the present invention include cyclopropyl, cyciobutyl, cydopentyl, cyclohexyl, cydoheptyl, and cyclooct l.

"Alkoxy" refers to a group containing an alkyl radical attached through an oxygen linking atom. The term "(C-|-C₆)alkoxy" refers to a straight- or branched-chain hydrocarbon radical having at ieast 1 and up to 6 carbon atoms attached through an oxygen linking atom. Exemplary "(Ci~C₄)-alkoxy" groups useful in the present invention include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-hutoxy, s-butoxy, and t-butoxy. Representative haioalkoxy include, but are not limited to dif!uoromethoxy (-OCHCF₂), trifluoromethoxy (-OCF₃), tetrafluoroethoxy (- OCF₂CHF₂) and the like.

"Alkylthio-" refers to a group containing an alkyl radical atoms attached through an sulfur linking atom. The term "(C1-C4)alkylthio-'^! refers to a straight- or branched-chain hydrocarbon radical having at Ieast 1 and up to 4 carbon atoms attached through a sulfur linking atom. Exemplary "(C1-C4)a!kyithio-" groups useful in the present invention include, but are not limited to, methylthio-, ethyithio-, n-propylthio-, Isopropyithio-, n-butylthio-, s-butylthio-, t-buty!thio- and the like.

Carbocyclic ring refers to a ring In which all ring atoms are carbon atoms, which may be aromatic or non-aromatic, fused or non-fused and the like. Examples of carbocyclic rings, may Include, but are not limited to cyc!oalky!s, such as

cyclopropane, cyciobutane, cyclopentane, cyclohexane and the like, aromatic or aryi rings, which include, but are not limited to rings such as benzene, naphthalene and the like, which include, but are not limited to fused ring compounds, such as 1 ,2,3,4- tetrahydronaphthaiene and the like.

"Cycioalkyloxy", "cycloaikylthio", "cyc!oa!ky!amlno" refers to a group containing a saturated carbocyclic ring atoms attached through an oxygen, nitrogen or sulfur linking atom, respectively.

"Aryi" represents a group or moiety comprising an aromatic, monovalent monocyclic or bicyciic hydrocarbon radical containing from 8 to 10 carbon ring atoms, which may be unsubstituted or substituted by one or more of the substituents defined herein, and to which may be fused one or more cycloalkyi rings, which may be unsubstituted or substituted by one or more substituents defined herein.

Representative aryi groups suitable for use in the present invention, may Include, but are not limited to phenyl, naphthaleny!, f!uorenyl, and the like.

Heteroatoms are defined as oxygen, nitrogen, sulfur and the like.

Heterocyclic groups may be heteroaryi or heterocycloalkyl groups. PU85023

Each monocyclic heterocyclic ring of the present invention has from 3 to 7 ring atoms and contains up to four heteroatoms. Monocyclic heterocyclic rings or fused heterocyclic rings include substituted aromatic and non-aromatics;

Each fused heterocyclic ring of the present invention optionally includes carbocyclic rings or heterocyclic rings;

"Heterocycioaikyl" represents a group or moiety comprising a monovalent monocyclic or bicycilc radical, which is saturated or partially unsaturated (non- aromatic), containing 3 to 10 ring atoms, which includes 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, and which may be unsubstituted or substituted by one or more of the substituents defined herein.

Illustrative examples of heterocycioalkyls include, but are not limited to, azetidiny!, pyrrol idyl (or pyrrolidinyl), piperidinyl, piperazinyl, morpholinyi, tetrahydro-2H-1 ,4- thiazinyl, tetrahydrofuryl (or tetrahydrofuranyl), dihydrofury!, oxazo!inyl, thlazolinyi, pyrazolinyl, tetrahydropyranyl, dihydropyranyl, 1 ,3-dioxoianyl, 1 ,3-dioxanyl, 1 ,4- dioxanyl, 1 ,3-oxathio!anyi, 1 ,3-oxathianyl, 1 ,3-dithianyl, azabicylo[3.2.1]octyi, azabicylo[3.3.1 ]nonyl, azabicylo[4.3.0]nonyl, oxabicyio[2.2.1]heptyi,

1 ,5,9-triazacyc!ododecyi and the like.

Generally, in the compounds of this invention, heterocycioaikyl groups are 5-membered and/or 6-membered heterocycioaikyl groups, such as pyrrolidyi (or pyrrolidinyl), tetrahydrofuryl (or tetrahydrofuranyl), tetrahydrothienyi, dlhydrofuryl, oxazolinyi, thiazoiinyl or pyrazolinyl, piperidyl (or piperidinyl), piperazinyl, morpholinyi, tetrahydropyranyl, dihydropyranyl, 1 ,3-dioxanyl, tetrahydro-2H-1 ,4-thiazinyl, 1 ,4- dioxanyl, 1 ,3-oxathianyl, and 1 ,3-dithianyl.

Additional examples of substituted heterocycioaikyl groups, which are partially saturated non-aromatic groups that are suitable In the present invention, may Include, but are not limited to: pyridin-4(1 H)-one, 3-hydroxy-1-methylpyridin-4(1 H)-one, 3,4- dihydroisoquinolin-1 (2H)-one, quinoiin-4(1 H)-one, 3-hydroxyquinolin-4(1 H)-one, 3- hydroxy-1 -methyiquinoiin-4(1 H)-one, 5-ch!oro-1 -ethy!-6,7-dihydroxyquinolin-4(1 H)-one, 5-chloro-6,7-dihydroxy-1-methylquinolin-4(1 H)-one, 1-ethyl-8-fluoro-6,7- dihydroxyquinolin-4(1 H)-one, 5-chloro-1 -ethyi-6,7-dihydroxyquinolin-4(1 H)-one, 1- ethyl-5-fluoro-6,7-dihydroxyquinoiin-4(1 H)-one, 1-ethy!-6-f!uoro-7,8-dihydroxyquinolin- 4(1 H)-one, 1-ethyi-7,8-dihydroxyquinolin-4(1 H)-one, 6,7-dihydroxy-1-isopropylquinolin- 4(1 H)-one, 1-ethyi-5,6-dihydroxyquinolin-4(1 H)-one, 5-chloro-1-cyclopropy!-6,7- dihydroxyquinolin-4(1 H)-one, 1-cyclopropyl-6,7-dihydroxyquinoiin-4(1 H)-one, 1-(tert- butyl)-6,7-dihydroxyquinolin-4(1 H)-one, 6,7-dihydroxy-1-methyiquinolin-4(1 H)-one, PU85023 cinnoiin-4(1 H)-one, 1-ethyi-6,7-di ydroxycinnolin-4(1 H)-one, 5-chloro-1-ethyl-6,7- dihydroxycinnoiin-4(1 H)-one, 1-ethyi-5-fiuoro-6,7-dihydroxycinnolin-4(1 H)-one, 1 -ethyl 6,7-dihydroxycinnoiin-4(1 H)-one and the like.

Additional examples of substituted heterocycloalkyl groups, which are non- aromatic that are suitable in the present Invention, may inciude, but are not limited to:

5-chloro-6_>7-bis((4-methoxybenzyi)oxy)-1-oxo-3_l4-dihydroisoquino!in-2(1 H)-yi;

5-hydroxy-1-methyl-4-oxo-1 ,4-dihydropyndine-2-carboxamido;

7-hydroxy-1 ,8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)-yl);

3-hydroxy-1-methyl-4-oxo-1 ,4-dihydroquinolin-6-yl;

3-hydroxy-1-methyl-4-oxo-1 ,4-dihydroquinoline-6-carboxamido;

1-ethy!-8,7-dlhydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yl;

1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido;

1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido;

3-hydroxy-1-methyl-4-oxo-1 ,4-dihydroquinoline-8-carboxamido;

1-ethyl-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yi;

5-ch loro- 1 -ethy I-6 , 7-d i h yd roxy-4-oxo- 1 ,4-d i hyd roq u i nol i ri-3-y I ;

1 -et y I -6 , 7-d i h yd roxy-4-oxo- 1 ,4-d i h yd roq u i no I i n -3-y I ;

5-chloro-1 -ethyi-8,7-dihydroxy-4~Qxo-1 ,4-dihydroquinoiine-3-carboxamido;

1 -ethyi-6,7-dihydroxy-4-oxo-1 ,4-di ydrocinnolin-3-yi;

1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido;

S-chloro-l -ethyl-BJ-dihydroxy^-oxo-l ^-dihydrocinnoline-S-carboxamido;

5-ch loro- 1 -ethy I-6 , 7-d i hyd roxy-4-oxo- 1 ,4-d i hyd roci n nol i n-3-y I ;

1 -ethyl-5-f I uoro-8 , 7-d I hyd roxy-4-oxo- 1 ,4-d i hyd roq u i nol I n-3-y Ϊ ;

5-fluoro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yi;

1 -ethyi-5-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido;

1 -ethyi-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yl;

1 -ethyi-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido;

1 -eth y I -7 , 8-d I h yd roxy-4~oxo- 1 ,4-d i h yd roq u i no I i n -3-y I ;

1 ~ethy!~7,8~dlhydroxy~4~oxo~1 ,4-dihydroquinoline-3-carboxamido;

1 ~ethy!~5,8~dlhydroxy~4~oxo~1 ,4-dihydroquinoline-3-carboxamido;

1 -(tert-butyl)-6,7-dihydroxy-4-oxo-1 ,4-dihydroqulnolin-3-yl;

1 -(tert-butyl)-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxamido;

1 -cyc!opropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-y!;

1 -cyc!opropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido;

5-chloro-1 -cyclopropyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl; PU85023

5-chloro-1 -cyclopropyi-67-di ydroxy-4-oxo-1 ,4-di ydroquinoiine-3-carboxamido; 6,7-dihydroxy-1 -methyl-4-oxo-1 ,4-dihydroquinoiin-3-yl;

6,7-dihydroxy-1 -isopropyl-4-oxo-1 ,4-dihydroquinoiin-3-yl

5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxarnido;

1-ethy!-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido;

1-ethy!-8-fluoro-6,7-di ydroxy-4-oxo-1 ,4-dihydroquino!in-3~yl;

5-chloro~6J-dihydroxy-1 ~meihy!-4~oxo~1 ,4~dihydroquinoliri~3-y! and the like.

"Heteroaryi" represents a group or moiety comprising an aromatic monovalent monocyclic or bicyclic radical, containing 5 to 10 ring atoms, including 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, which may be unsubstituted or substituted by one or more of the substituents defined herein. This term also encompasses bicyclic heteroeycl!c-aryi compounds containing an aryi ring moiety fused to a heterocycioaikyl ring moiety, containing 5 to 10 ring atoms, including 1 to 4 heteroatoms independently selected from nitrogen, oxygen and sulfur, which may be unsubstituted or substituted by one or more of the substituents defined herein. Illustrative examples of heteroaryls include, but are not limited to, thienyl, pyrrolyl, imldazo!yi, pyrazo!yl, furyl (or furanyl), isothiazoiy!, furazany!, isoxazo!yi, oxazolyl, oxadiazoiyl, thiazoiy!, pyridy! (or pyridinyl), pyrazinyl, pyrimldlnyl, pyrldazinyi, triazinyl, tetrazinyi, triazolyl, tetrazo!yi, benzo[bjthienyl, Isobenzofuryl, 2,3-dihydrobenzofuryi, chromenyi, chromanyl. indoliziny!, isolndoiyl, indolyi, indazoiyl, puriny!, isoquinolyi, quinoly!, phthalazinyl. naphthridinyl, quinzolinyl. benzothiazoiyl, benzimidazolyl, tetrahydroquinolinyl, cinnolinyl, pteridiny!, isothiazoiyl, carbazoly!, 1 ,2,3,4 tetrahydro Isoqulnolinyi and the like.

Generally, the heteroaryi groups present in the compounds of this invention are

5-membered and/or 6-memebred monocyclic heteroaryi groups. Selected 5- membered heteroaryi groups contain one nitrogen, oxygen or sulfur ring heteroatom, and optionally contain 1 , 2 or 3 additional nitrogen ring atoms. Selected 6-membered heteroaryi groups contain 1 , 2, 3 or 4 nitrogen ring heteroatoms. Selected 5- or 8~ membered heteroaryi groups include thienyl, pyrrolyl, imidazo!yi, pyrazolyl, furyl, Isothiazoiyl, furazanyl, isoxazoiyl, oxazoiy!, oxadiazoiyl, thiazo!yi, triazolyl, and tetrazolyi or pyridyi, pyrazinyl, pyrimldlnyl, pyrldazinyi, and triazinyl.

"Oxo" represents a double-bonded oxygen moiety; for example, if attached directly to a carbon atom forms a carbonyl moiety (C=0), or attached to an N or S forms oxides, N-oxides, sulfones or sulfoxides. PU85023

As used herein, the term "compound(s) of the invention" means a compound of Formulas (!) to (IX), respectively (as defined above) in any form, i.e., any salt or non- salt form (e.g., as a free acid or base form, or as a pharmaceutically acceptable salt thereof) and any physical form thereof (e.g., including non-solid forms (e.g., liquid or semi-solid forms), and solid forms (e.g., amorphous or crystalline forms, specific polymorphic forms, solvates, including hydrates (e.g., mono-, dl- and hemi- hydrates)), and mixtures of various forms.

As used herein, the term "optionally substituted" means that a group, such as, which may include, but Is not limited to alkyl, aryl, heteroary!, etc., may be

unsubstituted, or the group may be substituted with one or more substituent(s) as defined. In the case where groups may be selected from a number of alternative groups the selected groups may be the same or different.

The term Independently" means that where more than one substituent is selected from a number of possible substituents, those substituents may be the same or different.

The present invention relates to a compound of Formulas (I) to (IX), which definition referred herein includes, but are not limited to the following related sub- generic Formulas (II) and (IX),

The alternative definitions for the various groups and substituted groups of Formulas (I) to (IX), , respectively, or a pharmaceutically acceptable salt thereof, provided throughout the specification are intended to particularly describe each compound species disclosed herein, individually, as well as groups of one or more compound species. The scope of this invention includes any combination of these group and substituent group definitions.

The alternative definitions for the various groups and substituted groups of

Formulas (!) to (IX), , respectively, or a pharmaceutically acceptable salt thereof, provided throughout the specification are intended to particularly describe each compound species disclosed herein, individually, as well as groups of one or more compound species. The scope of this invention includes any combination of these group and substituent group definitions.

EMANTIOMERS. D!ASTE EQME S AND POLYMORPHS

The compounds according to Formulas (I) to (IX), respectively, or a

pharmaceutically acceptable salt thereof of the present invention may contain one or more asymmetric center (also referred to as a chiral center) and may, therefore, exist as individual enantiomers, diastereomers. or other stereoisomeric forms, or as mixtures PU85023 thereof. Chiral centers, such as chlral carbon atoms, may also be present in a substituent such as an alky! group. Where the stereochemistry of a chiral center present in Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof, or in any chemical structure illustrated herein, is not specified the structure is intended to encompass all individual stereoisomers and all mixtures thereof. Thus, compounds according to Formulas (I) to (IX), respectively, or a pharmaceuticaliy acceptable salt thereof, containing one or more chiral center may be used as racemic mixtures, enantiomericaliy enriched mixtures, or as enantiomericaliy pure individual stereoisomers.

individual stereoisomers of a compound according to Formulas (i) to (IX), respectively, or a pharmaceutically acceptable salt thereof .which contain one or more asymmetric center may be resolved by methods known to those skilled in the art. For example, such resolution may be carried out (1 ) by formation of diastereoisomer!c salts, complexes or other derivatives; (2) by selective reaction with a stereolsomer-spec!fic reagent, for example by enzymatic oxidation or reduction; or (3) by gas-liquid or liquid chromatography in a chiral environment, for example, on a chiral support such as silica with a bound chiral ligand or in the presence of a chiral solvent. The skilled artisan will appreciate that where the desired stereoisomer Is converted into another chemical entity by one of the separation procedures described above, a further step Is required to liberate the desired form. Alternatively, specific stereoisomers may be synthesized by asymmetric synthesis using optically active reagents, substrates, catalysts or solvents, or by converting one enantiomer to the other by asymmetric transformation. When a disclosed compound or its salt is named or depicted by structure, it is to be understood that the compound or salt, including solvates (particularly, hydrates) thereof, may exist in crystalline forms, non-crystalline forms or a mixture thereof. The compound or salt, or solvates (particularly, hydrates) thereof, may also exhibit polymorphism (i.e. the capacity to occur in different crystalline forms). These different crystalline forms are typically known as "polymorphs." It is to be understood that when named or depicted by structure, the disclosed compound, or solvates (particularly, hydrates) thereof, also include all polymorphs thereof. Polymorphs have the same chemical composition but differ in packing, geometrical arrangement, and other descriptive properties of the crystalline solid state. Polymorphs, therefore, may have different physical properties such as shape, density, hardness, deformability, stability, and dissolution properties. Polymorphs typically exhibit different melting points, IR spectra, and X-ray powder diffraction patterns, which may be used for identification. One of ordinary skill in the art will appreciate that different polymorphs may be produced, for example, by changing or adjusting the PU85023 conditions used in crystal!izing/recrysta!lizing the compound.

Because of their potentia! use in medicine, the salts of the compounds of

Formulas (!) to (!X), respectively, are preferably pharmaceutically acceptable salts.

Suitable pharmaceutically acceptable salts include those described by Berge, Bighley and Monkhouse J.Pharm.Sci (1977) 66, pp 1-19.

When a compound of the invention is a base (contain a basic moiety), a desired salt form may be prepared by any suitable method known in the art, including treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like, or with an organic acid, such as acetic acid, trifluoroacetic acid, maleic acid, succinic acid, mandeilc acid, fumaric acid, ma!onic acid, pyruvic acid, oxalic acid, glycolic acid, salicylic acid, pyranosidyi acid, such as glucuronic acid or galacturonic acid, alpha-hydroxy acid, such as citric acid or tartaric acid, amino acid, such as aspartic acid or glutamic acid, aromatic acid, such as benzoic acid or cinnamic acid, sulfonic acid, such as p-toluenesuifonic acid, methanesuifonic acid, ethanesulfonic acid or the like. Examples of pharmaceutically acceptable salts include sulfates, pyrosulfates, bisulfates, sulfites, bisuifites, phosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, capryiates, acrylates, formates, isobutyrates, caproates, heptanoates, propioiaf.es, oxalates, malonates succinates, su berates, sebacates, fumarates, maieates, butyne-1 ,4-dioates, hexyne-1 ,6- dioates, benzoates, chlorobenzoates, methyibenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phtha!ates, phenylacetates, phenylpropionates, phenyibutrates, citrates, lactates, y-hydroxybuty rates, g!ycoliates, tartrates mandelates, and sulfonates, such as xylenesu!fonates, methanesulfonates, propanesulfonates, naphthalene- 1 -sulfonates and naphtha!ene-2-sulfonates.

if an inventive basic compound Is isolated as a salt, the corresponding free base form of that compound may be prepared by any suitable method known to the art,

Including treatment of the salt with an Inorganic or organic base, suitably an inorganic or organic base having a higher pK than the free base form of the compound.

When a compound of the invention is an acid (contains an acidic moiety), a desired salt may be prepared by any suitable method known to the art, including

treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary, or tertiary), an a!kali metal or alkaline earth metal hydroxide, or the like, illustrative examples of suitable salts include organic salts derived from amino acids such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as ethylene diamine, dicyclohexyiamine, ethanolamine, piperidine, PU85023 morphollne, and piperazine, as well as inorganic saits derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum, and lithium.

Certain of the compounds of this Invention may form salts with one or more equivalents of an acid (if the compound contains a basic moiety) or a base (if the compound contains an acidic moiety). The present invention includes within its scope aii possible stoichiometric and non-stoichiometric salt forms.

Because the compounds of this invention may contain both acid and base moieties, pharmaceutically acceptable salts may be prepared by treating these compounds with an alkaline reagent or an acid reagent, respectively. Accordingly, this invention also provides for the conversion of one pharmaceutically acceptable salt of a compound of this invention, e.g., a hydrochloride salt, into another pharmaceutically acceptable salt of a compound of this invention, e.g., a sodium salt or a disodium salt.

Carboxy!ate functional groups of compounds of the present invention have coordinated mono or di-valent cations, where such cations may include, but are not limited to alkali metals, which may include, but are not limited to lithium (Li), sodium (Na), potassium, or mixtures thereof and the like.

Quarternary amine functional groups of compounds of the present invention, which are positively charged species, also may have coordinated anions, where such anions may include, but are not limited to halogens, which may Include, but are not limited to chlorides, fluorides, bromides, iodides and the like.

Compounds of Formulas (i) to (IX) of the present invention, also may form a zwitterion(s) (formerly called a dipolar ion), which Is a neutral molecule with a positive and a negative electrical charge (I.e., not dipoles) at different locations within that molecule. Zwltterions are sometimes also called inner salts.

SOLVATES

For solvates of the compounds of the invention, or salts thereof, that are In crystalline form, the skilled artisan will appreciate that pharrnaceuticaliy-acceptable solvates may be formed wherein solvent molecules are incorporated Into the crystalline lattice during crystallization. Solvates may involve nonaqueous solvents such as ethanoi, isopropanol, D SO, acetic acid, ethanolamine, and ethyl acetate, or they may involve water as the solvent that is Incorporated into the crystalline lattice. Solvates wherein water is the solvent that Is incorporated into the crystalline lattice are typically referred to as "hydrates." Hydrates include stoichiometric hydrates as well as compositions containing variable amounts of water. The invention includes all such solvates. PU85023

OBLITERATED COMPOUNDS

The invention also includes various deuterated forms of the compounds of Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof. Each available hydrogen atom attached to a carbon atom may be Independently replaced with a deuterium atom. A person of ordinary skill in the art will know how to synthesize deuterated forms of the compounds of Formulas (I) to (IX), respectively, or a

pharmaceutically acceptable salt thereof of the present Invention. For example, deuterated materials, such as a!kyl groups may be prepared by conventional techniques (see for example: methyl-dy-amine available from Aldrich Chemical Co., Milwaukee, Wl, Gat. No.489,689-2).

ISOTQPTES

The subject invention also includes isotopical!y-labeled compounds which are Identical to those recited In Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, fluorine, iodine and chlorine such as ³H, ¹ , '⁴C, ⁱ⁸F, ¹²³l or ^2DI.

Compounds of the present invention and pharmaceutically acceptable salts of said compounds that contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of the present invention. Isotopicaily labeled compounds of the present invention, for example those into which radioactive isotopes such as ^JH or ¹ C have been incorporated, are useful in drug and/or substrate tissue distribution assays. Trltiated, ie. ³H, and carbon-14, ie. ¹⁴C, isotopes are particularly preferred for their ease of preparation and detectabillty. ^{1 1}C and ⁱ⁵F isotopes are particularly useful In PET (positron emission tomography).

PURITY

Because the compounds of the present invention are intended for use in pharmaceutical compositions it will readily be understood that they are each preferably provided In substantially pure form, for example at least 60% pure, more suitably at least 75% pure and preferably at least 85%, especially at least 98% pure (% are on a weight for weight basis). Impure preparations of the compounds may be used for preparing the more pure forms used in the pharmaceutical compositions.

SYNTHETIC SCHEMES AND GENERAL METHODS OF PREPARATION

The compounds of Formulas (!) to (IX), respectively, or a pharmaceutically PU85023 acceptable salt thereof, may be obtained by using synthetic procedures illustrated In the Schemes below or by drawing on the knowledge of a skilled organic chemist.

The synthesis provided In these Schemes are applicable for producing

compounds of the invention having a variety of different R¹ and R² groups employing appropriate precursors, which are suitably protected if needed, to achieve compatibility with the reactions outlined herein. Subsequent deprotection, where needed, affords compounds of the nature generally disclosed. While the Schemes are shown with compounds only of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof, they are illustrative of processes that may be used to make the compounds of the invention.

intermediates (compounds used In the preparation of the compounds of the invention) may also be present as salts. Thus, In reference to Intermediates, the phrase "compound(s) of formula (number)" means a compound having that structural formula or a pharmaceutically acceptable salt thereof.

The present Invention also relates to processes for making compounds of

Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof.

The compounds of the present invention may be obtained by using synthetic procedures illustrated in Schemes below or by drawing on the knowledge of a skilled organic chemist.

The synthesis provided in these Schemes are applicable for producing

compounds of the invention as defined by Formulas (I) to (IX), respectively, or a

pharmaceutically acceptable salt thereof, respectively, having a variety of different functional groups as defined employing appropriate precursors, which are suitably protected if needed, to achieve compatibility with the reactions outlined herein.

Subsequent deprotection, where needed, affords compounds of the nature generally disclosed. While the Schemes shown with compounds only as defined therein, they are Illustrative of processes that may be used to make the compounds of the invention.

Intermediates (compounds used In the preparation of the compounds of the invention) aiso may be present as salts. Thus, In reference to Intermediates, the phrase "compound(s) of formula (number)" means a compound having that structural formula or a pharmaceutically acceptable salt thereof.

The compounds according to Formulas (i) to (IX), respectively, or a pharmaceutically acceptable salt thereof, or pharmaceutically acceptable salts thereof, are prepared using conventional organic syntheses. Suitable synthetic routes are depicted below in the following general reaction schemes. The skilled artisan will appreciate that if a substituent described herein is not compatible with the synthetic methods described herein, the substituent may be protected with a suitable protecting group that is stable to the reaction conditions. The protecting group may be removed at a suitable point in the reaction sequence to provide a desired intermediate or target compound. Suitable protecting groups and the methods for protecting and de-protecting different substituents using such suitable protecting groups are well known to those skilled in the art; examples of which may be found in T. Greene and P. Wuts, Protecting Groups in Chemical Synthesis (3rd ed.), John Wiley & Sons, NY (1999). In some instances, a substituent may be specifically selected to be reactive under the reaction conditions used. Under these circumstances, the reaction conditions convert the selected substituent into another substituent that is either useful as an intermediate compound or is a desired substituent in a target compound.

Synthetic Schemes

Scheme I

Scheme 1 represents a genera! scheme for the preparation of compounds of the present i

where: PU85023

X is N, or C-R^s.

R^a is hydrogen, chlorine or halogen;

R ' and R² each are hydrogen, (C1 -6)~a!kyi, or (CH₂)p-C(0)OR'⁸

where: p is an integer from 1 to 5

U is S or S-0

P^" is a counter anion of a quaternary amine;

R ' ³ , R '⁶, R^iS represent an ester residue such as a carboxyi-protecting group. R^{i 5} is a hydrogen or a carboxy protecting group. Such esters include those esters that are readily metabolized in the body to form a carboxyilc state. The aforementioned carboxyi-protecting group may be of any group as long as it can be protected and/or deprotected by a method described in Protective Groups in Organic Synthesis, Theodora W Green (John Wiley & Sons), or the like. Examples of suitable protecting groups may include, but are not limited to lower alkyl (e.g., methyl, ethyl, f-butyl), lower aikylcarbonyloxymethyl (e.g., pivaloy!), optionally substituted araikyl (e.g., benzyl, benzhydryl, phenethy!, p-methoxybenzyl, p-nitrobenzyl), sl!yl groups (f-butyldimethylsilyi, dlphenyl(f-butyl)sllyl), and the like.

R ' represent an amino-protecting group.

R '' is a hydrogen or an amino protecting group. Such amino-protecting groups Include those groups that are readily metabolized in the body to form amino. The aforementioned amino-protecting group may be of any group as long as it can be

protected and/or deprotected by a method described in Protective Groups in Organic Synthesis, Theodora W Green (John Wiley & Sons), or the like. Examples of suitable protecting groups, may include, but are not limited to may include lower alkoxycarbonyl (e.g., i-butoxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl), optionally

substituted aralkanoyl (e.g., benzoyl, p-nitrobenzoyl), acyl (e.g., formyl, chloroacetyl), and the like.

LG and Y as defined above represent leaving groups (for example, hydroxy, halogen (CI, Br, I), optionally substituted by such groups, which may include, but are not iimited to carbamoyioxy, acyloxy, methanesuifonyioxy, and toluenesulfonyloxy, etc.;. The following structure:

Is a moiety of Formulas (I) and (II) including a quaternary ammonium group moiety of 3- side chain: where each symbol is defined as above;

(1 ) Raw Maierials of the 7-Ss e C asrt: Synthesis of Compound CYSl First Step: PU85023

Compound (III) Is obtained by reacting N-hydroxyphtha!imide in the presence of Compound (I!) (LG is hydroxy) and a Mitsunobu reagent, or in the presence of Compound (Π) (LG is another leaving group) and a base (such as sodium hydroxide, sodium methoxide).

The amount of N-hydroxyphthailmide used is generally in the range from 1 to 5 molar equivalents, more specifically in the range from 1 to 2 molar equivalents, relative to Compound (Π).

Examples of reaction solvents, may include, but are not limited to ethers (e.g., dioxane, tetrahydrofuran, diethyl ether, fert-butyl methyl ether, diisopropyl ether), halogenated hydrocarbons(e.g., dichioromethane, chloroform, carbon tetrachloride), hydrocarbons (e.g., n-hexane, benzene, toluene), amides (e.g., formamide, N,N- dimethylformamide. Ν,Ν-dimethylacetamide, N-methylpyrroiidone), and the like, and mixed solvents and the like thereof.

A reaction temperature is in a range of, generally, about -50 to 100°C, preferably about -40 to 50^C'C, and more preferably about -30 to 0"C.

Secortd Step:

N-Methyihydrazine or hydrazine is added and reacted to Compound (ill) to provide Compound (IV).

The amount of N-methy!hydrazine or hydrazine used is in a range of about 1-10 molar equivalents, preferably 1-5 molar equivalents, more preferably 1-2 molar equivalents, relative to Compound (Hi).

Examples of reaction solvents, which may include, but are not limited to ethers (e.g., dioxane, tetrahydrofuran, diethyl ether, iert-butyl methyl ether, diisopropyl ether), esters (e.g., ethyl formate, ethyl acetate, Isopropyl acetate), halogenated

hydrocarbons(e.g., dichioromethane, chloroform, carbon tetrachloride), hydrocarbons (e.g., n-hexane, benzene, toluene), alcohols (e.g., methanol , ethanoi, isopropanol), amides (e.g., formamide, N,N-dimethy!formamide, Ν,Ν-dimethyiacetamide, N- methylpyrrolidone), ketones (e.g., acetone, methyl ethyl ketone), nitriies (e.g., MeCN, proplonltrile), dimethy!sulfoxide, water, and the like, and mixed solvents and the like thereof.

A reaction temperature is in a range of, generally, about 0 to 100°C, preferably about 0 to 50"C, more preferably about 10 to 30^GC.

Compound (IV) is added and reacted with Compound (V), which is commercially PU85023 available or obtained by a known method, to provide Compound (VI). (For example, it is described in Bioorganic & Medicinal Chemistry, vol.15, pp.6716-8732 (2007)).

N-Methylhydrazine or hydrazine is added and reacted with Compound (M l) to provide Compound (IV).

Examples of reaction solvents, which may include, but are not limited to ethers

(e.g. , dioxane, tetrahydrofuran , diethyl ether, ferf-butyl methyl ether, diisopropyi ether), esters (e.g ., ethyl formate, ethyl acetate, isopropyl acetate), ha!ogenated hydrocarbons (e.g. , dichioromethane, chloroform, carbon tetrachloride), hydrocarbons (e.g. n-hexane, benzene, toluene), alcohols (e.g. , methanol , ethanol , isopropanol). amides (e.g. , formamide, Ν ,Ν-dimethylformamide, N.N-dimethylacetamide, N-methylpyrrolidone), ketones (e.g ., acetone, methyl ethyl ketone), nitriles (e.g. , MeCN, propionitrile), dimethyisulfoxide, water and the like, and mixed solvents and the like thereof.

A reaction temperature is in a range of, generally, about 0 to 100°C, preferably about 0 to 50^!,C, more preferably about 10 to 30°C.

(2) 7-Amidation arid Formation of the 3-Side Chain; Synthesis of Compound (X¾ Fourth step {T-amidatsors reaction):

Compound (IX) is obtained by reacting Compound (VI) and Compound (VM), which are commercially available or synthesized according to methods described In a document (e.g. , Japanese Laid-Open Publication No. 60-231884, Japanese Laid-Open Publication No. 62-149682, etc. ). In this case, preferably, R¹³ and R ^t> are carboxy protecting groups, R¹⁴ is an amino protecting group, and R^{i 5} and R '⁷ are hydrogen.

The amount of Compound (VI) used is in a range of, generally, about 1 -5 moles, preferably 1 -2 mo!es, relative to 1 mole of Compound (VI I).

Examples of reaction solvents may include, but are not limited to ethers (e.g. , dioxane, tetrahydrofuran, diethyl ether, ferf-butyl methyl ether, diisopropyi ether), esters (e.g. , ethyl formate, ethyl acetate, isopropyl acetate), halogenated hydrocarbons(e.g. , dichioromethane, chloroform, carbon tetrachloride), hydrocarbons (e.g. , n-hexane, benzene, toluene), amides (e.g ., formamide, Ν,Ν-dimethylformamide, N,N- dimethylacetamide, N-methylpyrroiidone), ketones (e.g ., acetone, methyl ethyl ketone), nitriles (e.g. , MeCN , propionitrile), dimethyisulfoxide, water, and the like, and mixed solvents and the like thereof.

A reaction temperature is in a range of, generally, about -40 to 80°C, preferably about -20 to 50°C, more preferably about -10 to 30°C.

The above-described amidation reaction may be carried out after the carboxy moiety is converted into a reactive derivative (e.g ., which may include, but are not limited PU85023 to inorganic base saii(s), organic base sait(s), acid haiide(s), acid azide(s), acid anhydride(s), mixed acid anhydride(s), active amide(s), active ester(s), and active thioester(s) and the like). Examples of such inorganic bases may include, but are not limited to alkali metal (e.g., Na, K, and the like), alkali earth metal (e.g., Ca, Mg), and the like. Examples of organic bases suitable for use in the present invention, may include, but are not limited to trimethy!amine, triethy!amine, ferf-butyldimethylamine,

dibenzylmethylamine, benzy!dimethylamine, N-methyimorphoi!ne, di!sopropy!ethyiamine, and the like. Examples of acid haildes, may include, but are not limited to acid chlorides, acid bromides, and the like. Examples of mixed acid anhydrides, may include, but are not limited to mixed acid anhydrides of mono-aikyi carbonates, mixed acid anhydrides of aliphatic carboxylic acid, mixed acid anhydrides of aromatic carboxylic acid, mixed acid anhydrides of organic sulfonic acid, and the like. Examples of active amides, may include, but are not limited to amides with nitrogen-containing heterocyclic compound, and the like. Examples of active esters, may include, but are not limited to organic phosphoric esters (e.g., diethoxyphosphoric ester, diphenoxyphosphoric ester, and the like), p~n itrophenyl ester, 2,4-dinitrophenyl ester, cyanomethyi ester, and the like.

Examples of active thioesters include esters with aromatic heterocyclic thiol compound (e.g., 2-pyridyithioi esters), and the like.

Furthermore, in the above-described reaction, a suitable condensing agent may be used as desired. For example, hydrochloric acid salt of each of the following, which may include, but are not limited to 1 -dimethylaminopropyl-3-ethylcarbocliimide

(WSCD HCI), N.N'-dicyclohexylcarbodilmide, Ν,Ν'-carbonyidiimidazole, Ν,Ν'- thiocarbonyidiimidazole, N-ethoxycarbonyi-2-ethoxy-1 ,2-dihydroquinoline, phosphorus oxychloride, alkoxyaceiyiene, 2-chloromethylpyridinium iodide, 2-f!uoromethy!pyndinium Iodide, trifluoroacetic anhydride, and the like can be used as a condensing agent.

Fifth step ¾3-¾ide chain forming reaction!:

Compound (X) is obtained by reacting Compound (IX) and a corresponding tertiary amine. In this case, preferably, R '³ and R¹¹³ are carboxy protecting groups, and R¹⁴ is an amino protecting group.

An amount of the corresponding tertiary amine used is In a range of, generally, 1-

5 moles, preferably 1 -2 moles, relative to 1 mole of Compound (IX).

Examples of reaction solvents, may include, but are not limited to ethers (e.g., dioxane, tetrahydrofuran, diethyl ether, fert-butyl methyl ether, diisopropyl ether), esters (e.g., ethyl formate, ethyl acetate, isopropyl acetate), halogenated hydrocarbons(e.g., dichioromethane, chloroform, carbon tetrachloride), hydrocarbons (e.g., n-hexane, PU85023 benzene, toluene), amides (e.g., formamide, Ν,Ν-dimethylformamide, N,N- dimethylacetamide, N-methylpyrroiidone), ketones (e.g., acetone, methyl ethyl ketone), nitriles (e.g., MeCN, propionitrile), dimethyisulfoxide, water, and the like, and mixed solvents and the like thereof.

A reaction temperature generally is suitabiy in a range of about -20 to 60^CiC, more suitably is In a range of about -10 to 40°C, or most suitably is In a range of about 0 to 20°C.

Furthermore, a compound of which U is S in Compound (X) can be obtained by reducing the compound of which U is SO in Compound (X). Examples of reducing agents, may include, but are not limited to potassium iodide, acetyl chloride or phosphorous tribromide and the like.

{3} 3-side chairs formation and 7-amidation: Synthesis of Compound (X)

Sixt Step (3-side chain forming reaction}:

Compound (VIII) Is obtained by reacting Compound (Vi!) with a corresponding tertiary amine, in this case, preferably, R'⁶ is a carboxy protecting group, and R ' Is an amino protecting group.

An amount of the corresponding tertiary amine used is in a range of, generally, 1- 5 moles, preferably 1 -2 moles, relative to 1 mole of Compound (VII).

Examples of reaction solvents, may Include, but are not limited to (e.g., dioxane, tetrahydrofuran, diethyl ether, ierf-buty! methyl ether, diisopropyl ether), esters (e.g., ethyl formate, ethyl acetate, isopropy! acetate), halogenated hydrocarbons(e.g.,

dichioromethane. chloroform, carbon tetrachloride), hydrocarbons (e.g., n-hexane, benzene, toluene), amides (e.g., formamide, Ν,Ν-dimethylformamide, N,N- dimethylacetamide, N-methylpyrroiidone), ketones (e.g., acetone, methyl ethyl ketone), nitriles (e.g., MeCN, propionitrile), dimethyisulfoxide, water, and the like, and mixed solvents and the like thereof.

A reaction temperature generally is suitabiy In a range of about -20 to 60^t!C, more suitably is In a range of about -10 to 40°C, or most suitably is In a range of about 0 to 20 °C.

Both tertiary amine moieties used in the 3-side chain forming reactions of the fifth and the sixth steps can be obtained as a commercially available reagent, by a known method, and/or by a method described herein.

Seventh Step CT-amidatiori reactions:

Compound (X) is obtained by reacting Compound (VIII) and Compound (VI). In this case, preferably, R '³ and R ⁶ are carboxy protecting groups, R¹⁴ is an amino PU85023 protecting group, R¹⁵ and R ^? are hydrogen.

The amount of Compound (VI) generally used is suitably in a range of about 1-5 moles, more suitably In a range of about 1 to 2 moles, relative to 1 mole of Compound (VIII).

Examples of reaction solvents, which may include, but are not limited to ethers (e.g., dioxane, tetrahydrofuran, diethyl ether, ferf-butyl methyl ether, diisopropyi ether), esters (e.g., ethyl formate, ethyl acetate, isopropyl acetate), haiogenated

hydrocarbons(e.g., dichloromethane, chloroform, carbon tetrachloride), hydrocarbons (e.g., n-hexane, benzene, toluene), amides (e.g., formamide, N,N-dimethyiformamide, Ν,Ν-dimethy!acetamide, N-methy!pyrrolidone), ketones (e.g., acetone, methyl ethyl ketone), nitriles (e.g., MeCN, propionitriie), dimethylsuifoxide, water, and the like, and mixed solvents and the like thereof.

A reaction temperature generally is suitably in a range of about -40 to 80°G, more suitably in a range of about -20 to 50°C or most suitably In a range of about -10 to 30^t!C.

The above-described amidation reaction may be carried out after a carboxyl moiety is converted to a reactive derivative (e.g., which may include, but are not limited to Inorganic base salt, organic base salt, acid halide, acid azide, acid anhydride, mixed acid anhydride, active amide, active ester, and active thioester). Examples of suitable inorganic bases for use in the present invention include alkali metal (e.g., Na, K, and the like), alkali earth metal (e.g., Ca, Mg), and the like. Examples of suitable organic bases for use in the present invention, which may include, but are not limited to trimethylamine, triethylamine, iert-butyldimethylamine, dibenzylmethy!amine, benzyidimethyiamine, N- methy!morpho!ine, diisopropylethylamine, and the like. Examples of acid halides, which may include, but are not limited to acid chlorides, acid bromides, and the like. Examples of mixed acid anhydrides, which may include, but are not limited to mixed acid anhydrides of mono-alkyi carbonate, mixed acid anhydrides of aliphatic carboxyl ic acid, mixed acid anhydrides of aromatic carboxyilc acid, mixed acid anhydrides of organic sulfonic acid, and the like. Examples of active amides, which may include, but are not limited to amides with nitrogen-containing heterocyclic compound, and the like. Examples of active esters, which may include, but are limited to organic phosphoric esters (e.g., diethoxyphosphoric ester, diphenoxyphosphoric ester, and the like), p-nitrophenyl ester, 2,4-dinitrophenyl ester, cyanomethyl ester, and the like. Examples of active thioesters, which may include, but are not limited to esters with aromatic heterocyclic thiol compound (e.g., 2-pyridy!thiol esters), and the like. Examples of active thioesters, which may include, but are not limited to esters with aromatic heterocyclic thiol compound (e.g., 2-pyridylthio! esters), and the like. Furthermore, in the above-described reaction, a suitable condensing agent PU85023 may be used as desired. For example, hydrochloric acid salt of each of the following, which may include, but are not limited to 1-dimethylaminopropyl-3-ethylcarbodiimide (WSCD-HCI), Ν,Ν'-dicyc!ohexy!carbodiimide, Ν,Ν'-carbonyldlimidazole, Ν,Ν'- thiocarbonyidiimidazoie, N-eihoxycarbonyl-2~efhoxy-1 ,2-dihydroquinoline, phosphorus oxychloride, alkoxyacetylene, 2-chloromethylpyridiniurn iodide, 2-fluoromet ylpyridinium iodide, trifluoroacetic anhydride, and the like can be used as a condensing agent.

iori Reaction

Compound (!) is obtained by subjecting Compound (X) to a deprotection reaction with a method well-known to those skilled in the art.

Examples of reaction solvents, which may include, but are not limited to ethers (e.g., anisole, dioxane, tetrahydrofuran, diethyl ether, iert-butyl methyl ether, diisopropyl ether), esters (e.g., ethyl formate, ethyl acetate, n-butyl acetate), halogenated hydrocarbons(e.g., dlchloromethane, chloroform, carbon tetrachloride), hydrocarbons (e.g., n-hexane, benzene, toluene), amides (e.g., formamide, N,N-dlmethylformamlde, N,N-dimethylacetamide, N-methy!pyrrolidone), ketones (e.g., acetone, methyl ethyl ketone), nitriies (e.g., MeCN, propionitri!e), nitros (e.g., nitromethane, nltroethane, nitrobenzene), dimethyisuifoxiae, water, and the like. These solvents may be used alone or in a combination using two or more of such solvents.

A reaction temperature generally is suitably In a range of about -30 to 100°C, more suitably in a range of about 0 to 50°C, most suitably in a range of about 0 to 10°C.

As a suitable catalyst for use in the present invention, may include, but Is not limited to Lewis acid(s) (e.g., AICI_3l SnCI₄, TiCI₄), protonic acid(s) (e.g., HCI, HBr, H₂S0₄, HCOOH, CF₃C0₂H), and the like.

The obtained Compound (!) is or may be further chemically modified, and thereby an ester, or a compound of which an amino on the thiazoie ring at the 7-position thereof is protected, or a pharmaceutically acceptable salt, or a solvate thereof can be synthesized.

PHARMACEUTICAL CQMPOSmGNS. DOSAGE FORMS AMD REGiMENS

The present Invention relates to pharmaceutical compositions comprised of novel compounds of Formulas (!) to (IX), respectively, or a pharmaceutically

acceptable salt thereof and at least one pharmaceutically acceptable excipient(s).

The compounds of the invention will normally, but not necessarily, be

formulated into a pharmaceutical composition prior to administration to a patient.

Accordingly, the present invention is directed to pharmaceutical compositions or formulations, which comprise a compound or compound species of the present PU85023 invention and pharmaceutical!y-acceptable excipient(s). in particular, the present Invention also may relate to a pharmaceutical composition or formulation, which comprises a compound as defined by Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient(s), and optionally one or more other therapeutic ingredients.

The pharmaceutical compositions of the invention may be prepared and packaged in bulk form wherein an effective amount of a compound of the invention can be extracted and then given to the patient such as with powders, syrups, and solutions for injection. Alternatively, the pharmaceutical compositions of the invention may be prepared and packaged in unit dosage form. For oral application, for example, one or more tablets or capsules may be administered. A dose of the pharmaceutical composition contains at least a therapeutically effective amount of a compound of this Invention (i.e., a compound of Formulas (i) to (IX), respectively, or a pharmaceutically acceptable salt thereof, particularly a pharmaceutically acceptable salt, thereof).

When prepared in unit dosage form, the pharmaceutical compositions or formulations may contain from 1 mg to 1000 mg of a compound of this Invention.

The pharmaceutical compositions or formulations as defined herein typically contain one compound of the present Invention. However, in certain embodiments, the pharmaceutica! compositions may contain more than one compound of the present Invention. In addition, the pharmaceutical compositions of the present invention may optionally further comprise one or more additional pharmaceutically active compounds.

As used herein, "pharmaceuticaily-acceptabie excipient" means a material, composition or vehicle involved In giving form or consistency to the composition. Each excipient must be compatible with the other Ingredients of the pharmaceutical composition when commingled such that interactions which would substantially reduce the efficacy of the compound of the Invention when administered to a patient and Interactions which would result In pharmaceutical compositions that are not pharmaceuticaily-acceptabie are avoided. In addition, each excipient must of course be of sufficiently high purity to render it pharmaceuticaily-acceptabie.

Suitable pharmaceuticaily-acceptabie excipients will vary depending upon the particular dosage form chosen. In addition, suitable pharmaceuticaily-acceptabie excipients may be chosen for a particular function that they may serve in the composition.

For example, certain pharmaceuticaily-acceptabie excipients may be chosen for their ability to facilitate the production of uniform dosage forms. Certain PU85023 pharmaceutica!!y-aeceptab!e excipients may be chosen for their ability to facilitate the production of stable dosage forms. Certain pharmaceutical!y-acceptable excipients may be chosen for their ability to facilitate the carrying or transporting the compound or compounds of the invention once administered to the patient from one organ, or portion of the body, to another organ, or portion of the body. Certain pharmaceuticai!y- acceptable excipients may be chosen for their ability to enhance patient compliance. Moreover, pharmaceutical compositions, formulations, dosage forms, and the like, etc. may conveniently be presented in unit dosage form and may be prepared by any of the methods well known in the art of pharmacy.

All methods include the step of bringing the active ingredient into association with the carrier which constitutes one or more accessory ingredients. In general the formulations are prepared by uniformly and intimately bringing into association the active ingredient with liquid carriers or finely divided solid carriers or both and then, If necessary, shaping the product into the desired formulation.

Suitable pharmaceuticai!y-aeceptab!e excipients include the following types of excipients: diluents, fillers, binders, disintegrants, lubricants, glidants, granulating agents, coating agents, wetting agents, solvents, co-solvents, suspending agents, emuls!fiers, sweeteners, flavoring agents, flavor masking agents, coloring agents, anti- caking agents, humectants, chelating agents, plasticizers, viscosity increasing agents, antioxidants, preservatives, stabilizers, surfactants, and buffering agents. The skilled artisan will appreciate that certain pharmaceutieai!y-acceptable excipients may serve more than one function and may serve alternative functions depending on how much of the excipient is present in the formulation and what other ingredients are present in the formulation.

Skilled artisans possess the knowledge and skill In the art to enable them to select suitable pharmaceutica!ly-acceptable excipients in appropriate amounts for use In the invention, in addition, there are a number of resources that are available to the skilled artisan which describe pharmaceutically-acceptable excipients and may be useful In selecting suitable pharmaceutically-acceptable excipients. Examples include Remington's Pharmaceutical Sciences (Mack Publishing Company), The Handbook of Pharmaceutical Additives (Gower Publishing Limited), and The Handbook of

Pharmaceutical Excipients (the American Pharmaceutical Association and the Pharmaceutical Press).

The compounds of the invention and the pharmaceutically-acceptable excipient or excipients will typically be formulated into a dosage form adapted for administration PU85023 to the patient by the desired route of administration.

With regard to the present invention, conventional dosage forms include those adapted for (1 ) oral administration such as tablets, capsules, capiets, pi!ls, troches, powders, syrups, elixirs, suspensions, solutions, emulsions, sachets, and cachets; (2) parenteral administration such as sterile solutions, suspensions, and powders for reconstitution; (3) transdermal administration such as transdermal patches; (4) rectal administration such as suppositories; (5) inhalation such as aerosols and solutions; and (6) topical administration such as creams, ointments, lotions, solutions, pastes, sprays, foams, and gels.

The pharmaceutical compositions or formulations of the invention are prepared using techniques and methods known to those skilled in the art. Some of the methods commonly used in the art are described in Remington's Pharmaceutical Sciences (Mack Publishing Company).

in general, pharmaceutical compositions of the present invention are prepared using conventional materials and techniques, such as mixing, blending and the like.

The term "active agent" Is defined for purposes of the present Invention as any chemical substance or composition of the present invention, which can be delivered from the device into an environment of use to obtain a desired result.

The percentage of the compound in compositions can, of course, be varied as the amount of active in such therapeutically useful compositions is such that a suitable dosage will be obtained.

In one aspect, the present invention is directed to a pharmaceutical composition comprising a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.

In another aspect, the present invention is directed to a pharmaceutical composition comprising a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.

In another aspect, the present invention is directed to a pharmaceutical composition comprising a compound or compound species or a pharmaceutically acceptable salt thereof of the present invention as defined herein and one or more pharmaceutically acceptable excipients.

It will be appreciated that the actual preferred dosages of the compounds being used in the compositions of this invention will vary according to the particular composition formulated, the mode of administration, the particular site of PU85023 administration and the host being treated.

The active compounds of the present invention may be orally administered, for example, with an inert diluent, or with an assimilable edible carrier, or they can be enclosed in hard or soft shell capsules, or they can be compressed into tablets, or they can be incorporated directly with the food of the diet, etc,

in one aspect, the invention is directed to a solid oral dosage form such as a tablet or capsule comprising a safe and effective amount of a compound of the invention and a diluent or filler. Suitable diluents and fillers include lactose, sucrose, dextrose, mannitol, sorbitol, starch (e.g. corn starch, potato starch, and pre-gelatinized starch), cellulose and its derivatives (e.g. microcrystailine cellulose}, calcium sulfate, and dibasic calcium phosphate. The oral solid dosage form may further comprise a binder. Suitable binders include starch (e.g. corn starch, potato starch, and pre-geiatinized starch), gelatin, acacia, sodium alginate, aiginic acid, tragacanth, guar gum, povidone, and cellulose and Its derivatives (e.g. microcrystailine cellulose). The ora! so!id dosage form may further comprise a disintegrant. Suitable disintegrants Include crospovidone, sodium starch giycolate, croscarmeiose, aiginic acid, and sodium carboxymethyl cellulose. The oral solid dosage form may further comprise a lubricant. Suitable lubricants include stearic acid, rnagnesuim stearate, calcium stearate, and talc.

Where appropriate, dosage unit formulations for ora! administration can be microencapsulated. The composition can a!so be prepared to prolong or sustain the release as for example by coating or embedding particulate material In polymers, wax or the like.

The compounds of the invention may also be coupled with soluble polymers as targetable drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymer,po!yhydroxypropy!methacrylamide-phenoi, polyhvdroxyethyl aspartamide phenol, or polyethyleneoxidepolylysine substituted with palmltoy! residues. Furthermore, the compounds of the Invention may be coupled to a class of biodegradable polymers useful in achieving controlled release of a drug, for example, poiyiactic acid, poiepsilon caproiactone, poiyhydroxy butyric acid, poiyorthoesters, poiyaceta!s, polydihydropyrans, polycyanoacrylates and cross-linked or amphipathic block copolymers of hydrogeis.

METHODS OF TREATMENT

The present Invention also relates to methods for treating bacteria! infections, which comprises administering to a subject in need thereof an effective amount of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof or a corresponding pharmaceutical composition. PU85023

As used herein, "patient" refers to a human or other mammal.

Methods of Use and Treatment of Diseases

In one aspect, a compound of Formulas (I) to (IX), respectively, or a

pharmaceutically acceptable salt or corresponding pharmaceutical compositions of the present invention have a wide antimicrobial activity spectrum, and may be used for prevention or therapy against a variety of diseases caused by causative bacteria In a variety of mammals including humans, for example, airway Infectious diseases, urinary system infectious diseases, resi iratory system infectious diseases, sepsis, nephritis, cholecystitis, oral cavity infectious diseases, endocarditis, pneumonia, bone marrow membrane myelitis, otitis media, enteritis, empyema, wound infectious diseases, opportunistic infection and the like.

Compounds of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceutical compositions of the present invention exhibit high antimicrobial activity in particular against Gram negative bacteria, preferably, Gram negative bacteria of enterobacteria (£. co!i, Klebsiella, Serrat a, Enterohacier, Citrobacter, Morganella, Providencia, Proteus and the like), Gram negative bacteria colonized in respiratory system (Haemophilus, Moraxeila and the like), and Gram negative bacteria of glucose non fermentation (Pseudomonas aeruginosa, Pseudomonas other than P, aeruginosa, Stenotrophomonas, Burkholderia, Acinetobacter and the like).

Compounds of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof of the present invention are stable against beta-!actamase belonging to classes A, B, C and D which beta-!actamase is produced by these Gram negative bacteria, and have high antimicrobial activity against a variety of beta-lactam drug resistant Gram negative bacteria, such as extended spectrum β-iactamase (ESBL) producing bacteria and the like. These are extremely stable against metai!o-beta- lactamase belonging to Class B including In particular IMP type, VIM type, L-1 type and the like. Thus, these are effective against a variety of beta-lactam drug resistant Gram negative bacteria including Cephem and Carbapenem. Moreover, the compounds of the subject invention has antimicrobial activity against Gram positive bacteria including methicii!!n-res!stant Staphylococcus aureus ( RSA), penicillin-resistant Streptococcus pneumoniae (PRSP), and the like. Still more preferable compounds have features regarding kinetics in the body, such as blood concentration in which such is highly bioavailable, long duration of effects, and /or significant tissue migration. More preferable compounds are safe In terms of side effects. More preferable compounds have high water solubility, and thus preferable as an injecting drug, in particular. PU85023

Compounds of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceutical compositions may be administered parenterally or orally as an injecting agent, capsules, tablets, and granules, and

preferably, administered as an injecting agent. Amounts to be administered may usually be, per 1 kg of body weight of a patient or animal, about 0.1 to 100 mg/day, preferably, about 0.5 to 50 mg/day, If desired, divided into 2-4 times per day. Carriers when used as an injecting agent Is for example, distilled water, saline and the like, and base and the like may be used for pH adjustment.

When used as capsules, granules or tablets, carriers may be known excipients (e.g., starch, lactose, sucrose, calcium carbonate, calcium phosphate and the like}, binders (e.g., starch, acacia gum, carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose, and the like), lubricants (e.g., magnesium stearate, talc and the like), and the like.

Still compounds of the present invention have features regarding kinetics in the body, such as blood concentration In which such is highly bioavailable, long duration of effects, and /or significant tissue migration. More preferable compounds are safe in terms of side effects. More preferable compounds have high water solubility, and thus

preferable as an injecting drug, in particular.

Suitable compounds of Formulas (i) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceutical compositions of the present Invention are useful in treatment of infections caused by causative bacteria in a variety of mammals including humans, which include, but are not limited to infectious diseases, urinary system infectious diseases, reslpiratory system infectious diseases, sepsis, nephritis, cholecystitis, oral cavity infectious diseases, endocarditis, pneumonia, bone marrow membrane myelitis, otitis media, enteritis, empyema, wound infectious diseases, opportunistic infection and the like

Suitably the compounds of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceutical compositions of the present invention are useful in the treatment of bacterial infections, more particularly gram negative bacterial infections casued by:

Gram negative bacteria of enterobacteria, which include, but are not limited to E. coli, Klebsiella, Serratia, Enterobacter, Citrobacter, Morgane!ia, Providencia, Proteus and the like;

Gram negative bacteria colonized in respiratory system, which include, but are not limited to Haemophilus, Moraxeila and the like; and

- 80 - PU85023

Gram negative bacteria of glucose non fermentation, which include, but are not limited to Pseudornonas aeruginosa, Pseudomonas other than P. aeruginosa, Stenotrophomonas, Burkhoideria, Acinetobacter and the like.

The compounds of Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceuticai compositions of the present invention also are stable against beta-!actamase belonging to classes A, B, C and D which beta-!actamase is produced by these Gram negative bacteria, and have high antimicrobial activity against a variety of beta-lactam drug resistant Gram negative bacteria, such as ESBL producing bacteria and the like. These are extremely stable against metallo-beta-lactamase belonging to Class B including in particular IMP type, VIM type, L-1 type and the like. Thus, these are effective against a variety of beta-lactam drug resistant Gram negative bacteria including Cephem and Carbapenem.

Suitably the compounds of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceutical compositions of the present Invention have antimicrobial activity against Gram positive bacteria including methiciilin- resistant Staphylococcus aureus (MRSA), penicillin-resistant Streptococcus pneumoniae (PRSP), and the like.

Still compounds of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceutical compositions of the present Invention have features regarding kinetics in the body, such as blood concentration in which such is highly bioavailable, long duration of effects, and /or significant tissue migration. More preferable compounds are safe in terms of side effects.

Compounds of Formulas (i) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or pharmaceutical compositions of the present invention have high water solubility, and thus preferable as an injecting drug, In particular.

The present Invention specifically relate to methods for the treatment infectious diseases inciuding bacterial Infections, comprise administering an effective amount of a compound according to Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and/or corresponding pharmaceutical compositions to a patient in need thereof.

One embodiment of the present invention provides for a method for treating a bacteria! infection, which comprises administering a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof.

Another embodiment of the present invention provides for a method of treating a bacteria! infection, which comprises administering a pharmaceutical composition

- 81 - PU85023 comprising a compound of Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable adjuvant, carrier or excipient.

In another embodiment of the present invention provides for a method of treating a bacterial infection In humans comprising administration of a compound or compound species or a pharmaceutically acceptable salt thereof as defined herein.

In one aspect, the present invention relates to a method of treating a bacterial infection comprising administering a therapeutically effective amount of a compound of Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human In need thereof.

In another aspect, the present invention relates to a method of treating a bacterial infection, where the bacterial infection is caused by Gram negative bacte ia.

In another aspect, the present invention relates to a method of treating a bacterial Infection, where the Gram negative bacteria selected from Gram negative bacteria of enterobacteria, Gram negative bacteria colonized In respiratory, Gram negative bacteria of glucose non fermentation or β-iactam drug resistant Gram negative bacteria.

In another aspect, the present invention relates to a method of treating a bacterial infection, where:

the Gram negative bacteria of enterobacteria selected from E. coii, Klebsiella, Serratia, Enterobacter. Citrobacter, organeila, Providencia or Proteus;

the Gram negative bacteria colonized in respiratory system selected from

Haemophilus or oraxe!la;

the Gram negative bacteria of glucose non fermentation selected from

Pseudomonas aeruginosa, Pseudomonas other than P. aeruginosa, Stenotrophomonas or Burkholderla, Aclnetobacter; and

the beta-iactam drug resistant Gram negative bacteria is selected from ESBL producing bacteria.

In another aspect, the present invention relates to a method, where the bacterial Infection Is an airway infection, urinary system Infection, resipiratory system infection, sepsis infection, nephritis, cholecystitis, oral cavity infection, endocarditis, pneumonia, bone marrow membrane myelitis, otitis media, enteritis, empyema, wound infection or an opportunistic infection .

In another aspect, the present invention relates to a method of treating a bacterial Infection comprising administering a therapeutically effective amount of a compound of

- 82 - PU85023

Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human In need thereof.

In another aspect, the present Invention relates to a method for treating a gram- negative infection comprising administering a therapeutically effective amount of a compound of of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human In need thereof.

In another aspect, the present invention relates to a method for treating a gram- negative infection comprising administering a therapeutically effective amount of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human In need thereof.

in another aspect, the present invention relates to a method for Inhibiting activity of UDP-3-0-(R-3-hydroxydecanoyl)-N-acetylgiucosamine deaceiyiase (LpxC) comprising administering a therapeutically effective amouni of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof. in another aspect, the present invention relates to a method for treating antimicrobial activity against Gram positive bacteria comprising administering a therapeutically effective amount of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof.

In another aspect, the present invention relates to a method for treating antimicrobial activity against Gram positive bacteria, where the Gram positive bacteria is selected from methicillin-reslstant Staphylococcus aureus ( RSA) or penicillin-resistant Streptococcus pneumoniae (PRSP).

In another aspect, the present invention relates to a method for treating methicillin-reslstant Staphylococcus aureus (MRSA), comprising administering a therapeutically effective amount of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof.

In another aspect, the present invention relates to a method for treating penicillin- resistant Streptococcus pneumoniae (PRSP), comprising administering a therapeutically effective amount of a compound of Formulas (I) to (IX), respectively, or a

pharmaceutically acceptable salt thereof to a human in need thereof.

The present Invention specifically relate to methods for the treatment infectious diseases including bacterial Infections, comprise administering an effective amount of a pharmaceutical composition comprising a compound according to Formulas (!) to (IX), respectively,, or a pharmaceutically acceptable salt thereof, to a patient in need thereof.

One embodiment of the present invention provides for a method for treating a

- 83 - PU85023 bacteria! infection, which comprises administering a pharmaceutical composition comprising compound of Formulas (I) to (IX), respectively, or a pharmaceuticaiiy acceptable salt thereof.

Another embodiment of the present invention provides for a method of treating a bacteria! infection, which comprises administering a pharmaceutical composition comprising a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable excipient.

In another embodiment of the present invention provides for a method of treating a bacteria! infection in humans comprising administration of a pharmaceutical composition comprising a compound or compound species or a pharmaceutically acceptable salt thereof as defined herein.

In one aspect, the present invention relates to a method of treating a bacterial Infection comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof.

In another aspect, the present invention relates to a method of treating a bacterial infection, where the bacterial infection is caused by Gram negative bacteria.

In another aspect, the present invention relates to a method of treating a bacterial Infection, where the Gram negative bacteria selected from Gram negative bacteria of enterobacteria, Gram negative bacteria colonized In respiratory, Gram negative bacteria of glucose non fermentation or β-lactam drug resistant Gram negative bacteria.

the Gram negative bacteria of enterobacteria selected from E. coii, Klebsiella, Serratia, Enterobacter, Cltrobacter, organeila, Providencia or Proteus;

the Gram negative bacteria colonized in respiratory system selected from

Haemophilus or Moraxella;

the Gram negative bacteria of glucose non fermentation selected from

Pseudomonas aeruginosa, Pseudomonas other than P. aeruginosa, Sienoirophomonas or Burkholderia, Acinetobacter; and

In another aspect, the present invention relates to a method, where the bacterial Infection Is an airway infection, urinary system Infection, resipiratory system infection, sepsis infection, nephritis, cholecystitis, oral cavity infection, endocarditis, pneumonia,

- 84 - PU85023 bone marrow membrane myelitis, otitis media, enteritis, empyema, wound infection or an opportunistic infection .

In another aspect, the present invention relates to a method of treating a bacterial infection comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formulas (!) to (IX), respectively, or a

pharmaceutically acceptable salt thereof to a human in need thereof.

In another aspect, the present invention relates to a method for treating a gram- negative infection comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof.

in another aspect, the present invention relates to a method for Inhibiting activity of UDP-3-0— (R-3-hydroxydecanoyi)-N-acetylglucosamine deacetyiase (LpxC) comprising administering a therapeutically effective amount of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof. in another aspect, the present invention relates to a method for treating antimicrobial activity against Gram positive bacteria comprising administering a therapeutically effective amount of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof.

In another aspect, the present invention relates to a method for treating antimicrobial activity against Gram positive bacteria, where the Gram positive bacteria is selected from methiciilin-resistant Staphylococcus aureus ( RSA) or penicillin-resistant Streptococcus pneumoniae (PRSP).

In another aspect, the present invention relates to a method for treating methiciilin-resistant Staphylococcus aureus (MRSA), comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human In need thereof.

In another aspect, the present invention relates to a method for treating penicillin- resistant Streptococcus pneumoniae (PRSP), comprising administering a therapeutically effective amount of a pharmaceutical composition comprising a compound of Formulas (!) to (IX), respectively, or a pharmaceutically acceptable salt thereof to a human in need thereof.

As used herein, "infectious disease" refers to any disease characterized by the presence of a microbial infection, such as a bacterial infection.

As used herein, "treat" in reference to a condition means: (1 ) to ameliorate or PU85023 prevent the condition or one or more of the bioiogicai manifestations of the condition, (2) to interfere with (a) one or more points in the bioiogicai cascade that ieads to or is responsible for the condition or (b) one or more of the bioiogicai manifestations of the condition, (3) to ai!eviate one or more of the symptoms or effects associated with the condition, or (4) to slow the progression of the condition or one or more of the bioiogicai manifestations of the condition.

As indicated above "treatment" of a condition includes prevention of the condition. The skilled artisan wl!l appreciate that "prevention" is not an absolute term. In medicine, "prevention" is understood to refer to the prophylactic administration of a drug to substantially diminish the !ikelihood or severity of a condition or biological manifestation thereof, or to delay the onset of such condition or biological manifestation thereof.

As used herein, "effective amount" in reference to a compound of the invention means an amount of the compound sufficient to treat the patient's condition, but low enough to avoid serious side effects (at a reasonable benefit/risk ratio) within the scope of sound medical judgment. An effective amount of a compound will vary with the particular compound chosen (e.g., consider the potency, efficacy, and half-life of the compound); the route of administration chosen; the condition being treated; the severity of the condition being treated; the age, size, weight, and physical condition of the patient being treated; the medical history of the patient being treated; the duration of the treatment; the nature of concurrent therapy; the desired therapeutic effect; and like factors, and can be routinely determined by the skilled artisan.

ADMINISTRATION

Treatment regimen for the administration of compounds of Formulas (!) to (iX), respectively, or pharmaceutically acceptable salts thereof or corresponding

pharmaceutical compositions of the present invention also may be determined readily by those with ordinary skill in art.

The quantity of the compound, pharmaceutical composition, or dosage form of the present invention administered may vary over a wide range to provide in a unit dosage in an effective amount based upon the body weight of the patient per day to achieve the desired effect and as based upon the mode of administration.

The scope of the present Invention inc!udes all compounds, pharmaceutical compositions, or eontroi!ed-re!ease formulations or dosage forms, which is contained in an amount effective to achieve its intended purpose. VVhl!e individual needs vary, determination of optimal ranges of effective amounts of each component is within the skii! of the art.

- 86 - PU85023

Compounds of Formulas (I) to (IX), respectively, or pharmaceutically

acceptable salts thereof or corresponding pharmaceutical compositions of the present Invention may be administered by any suitable route of administration, including both systemic administration and topical administration. Systemic administration Includes oral administration, parenteral administration, transdermal administration, rectal administration, and administration by inhalation.

Parenteral administration refers to routes of administration other than enteral, transdermal, or by inhalation, and is typically by injection or infusion. Parenteral administration includes intravenous, intramuscular, and subcutaneous injection or infusion.

inhalation refers to administration into the patient's lungs whether inhaled through the mouth or through the nasal passages. In one aspect, pharmaceutical compositions, formulations, dosages, dosage forms or dosing regimens of the present Invention are adapted for administration by inhalation.

Topical administration Includes application to the skin as well as Intraocular,

Intravaginai, and intranasal administration.

Compounds of Formulas (!) to (IX), respectively, or pharmaceutically

acceptable salts thereof or corresponding pharmaceutical compositions of the present Invention may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. For example, doses may be administered one, two, three, or four times per day. Doses may be administered until the desired therapeutic effect is achieved or

Indefinitely to maintain the desired therapeutic effect.

Suitable dosing regimens for compounds of Formulas (!) to (IX), respectively, or pharmaceutically acceptable salts thereof or corresponding pharmaceutical compositions of the present Invention depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half-life, which can be determined by the skilled artisan. In addition, suitable dosing regimens, including the duration such regimens are administered, for a compound of the invention depend on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient being treated, the nature of concurrent therapy, the desired therapeutic effect, and like factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an Individual patient's response to the dosing regimen or over time as Individual patient needs change.

- 87 - PU85023

In another aspect, the invention is directed to a liquid oral dosage form. Oral liquids such as solution, syrups and elixirs can be prepared in dosage unit form so that a given quantity contains a predetermined amount of a compound of the Invention. Syrups can be prepared by dissolving the compound of the invention in a suitably flavored aqueous solution, while elixirs are prepared through the use of a non-toxic alcoholic vehicle. Suspensions can be formulated by dispersing the compound of the invention In a non-tox!e vehicle. Solubiiizers and emuisifiers such as ethoxylated isostearyi alcohols and poiyoxy ethylene sorbitol ethers, preservatives, flavor additive such as peppermint oil or natural sweeteners or saccharin or other artificial sweeteners, and the like can also be added.

in another aspect, the invention is directed to parenteral administration.

Pharmaceutical compositions adapted for parenteral administration include aqueous and non-aqueous sterile injection solutions which may contain anti-oxldants, buffers, bacteriostatic and solutes which render the formulation Isotonic with the blood of the Intended recipient: and aqueous and non-aqueous sterile suspensions which may include suspending agents and thickening agents. The compositions may be presented in unit- dose or multi-dose containers, for example sealed ampoules and vials, and may be stored In a freeze-dried (iyophilized) condition requiring only the addition of the sterile liquid carrier, for example water for injections, immediately prior to use. Extemporaneous Injection solutions and suspensions may be prepared from sterile powders, granules and tablets.

Typical dally dosages may vary depending upon the particular route of administration chosen. Typical daily dosages for oral administration, to a human weighing approximately 70kg, would range from 7mg to 7g. suitably 3.5mg to 3.5g of a compound of the Invention a day.

Compounds of Formulas (I) to (IX), respectively, or pharmaceutically acceptable salts thereof or corresponding pharmaceutical compositions of the present invention may be administered parenteraily or orally as an injecting agent, capsules, tablets, and granules, and preferably, administered as an injecting agent. Amounts to be

administered may usually be, per 1 kg of body weight of a patient or animal, about 0.1 to 100 mg/day, preferably, about 0.5 to 50 mg/day, if desired, divided into 2-4 times per day. Carriers when used as an injecting agent is for example, distilled water, saline and the like, and base and the like may be used for pH adjustment. When used as capsules, granules or tablets, carriers may be known excipients (e.g., starch, lactose, sucrose, calcium carbonate, calcium phosphate and the like), binders (e.g., starch, acacia gum,

- 88 - PU85023 carboxymethy! cellulose, hydroxypropyl cellulose, crystalline cellulose, and the like), lubricants (e.g., magnesium stearate, talc and the like), and the like.

For all methods of use disclosed herein for the compounds of Formulas (I) to (IX), the daily oral dosage regimen will preferabiy be from about 0.05 to about 80 mg/kg of total body weight, preferably from about 0.1 to 30 mg/kg, more preferabiy from about 0.5 mg to 15mg/kg, administered in one or more daily doses. For example, the daily parenteral dosage regimen about 0.1 to about 80 mg/kg of total body weight, preferabiy from about 0.2 to about 30 mg/kg, and more preferably from about 0.5 mg to 15mg/kg, administered In one or more daily doses. The daily topical dosage regimen will preferabiy be from 0.01 mg to 150 mg, administered one to four times daily. The daily inhalation dosage regimen will preferably be from about 0.05 microgram/kg to about 5 mg/kg per day, or from about 0.2 microgram/kg to about 20 microgram/kg, administered in one or more daily doses.

It will also be recognized by one of skill in the art that the optimal quantity and spacing of individual dosages of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof will be determined by the nature and extent of the condition being treated, the form, route and site of administration, and the

particular patient being treated, and that such optimums can be determined by conventional techniques. It will also be appreciated by one of skill In the art that the optimal course of treatment, i.e., the number of doses of a compound of Formulas (I) to (IX), respectively, or a pharmaceutically acceptable salt thereof given per day for a defined number of days, can be ascertained by those skilled in the art using

conventional course of treatment determination tests.

The amount of a compounds of Formulas (I) to (IX), respectively, or

pharmaceutically acceptable salts thereof or corresponding pharmaceutical

compositions of the present Invention which is required to achieve a therapeutic effect will, of course, vary with the particular compound, the route of administration, the subject under treatment, and the particular disorder or disease being treated.

Suitable dosing regimens for a compound of the invention depend on the pharmacokinetic properties of that compound, such as absorption, distribution, and half-life, which can be determined by the skilled artisan, in addition, suitable dosing regimens, Including the duration such regimens are administered, for a compound of the invention depend on the condition being treated, the severity of the condition being treated, the age and physical condition of the patient being treated, the medical history of the patient to be treated, the nature of concurrent therapy, the desired therapeutic

- 89 - PU85023 effect, and iike factors within the knowledge and expertise of the skilled artisan. It will be further understood by such skilled artisans that suitable dosing regimens may require adjustment given an individual patient's response to the dosing regimen or over time as individual patient needs change.

Additionally, the compounds of the present invention may be administered as prodrugs. As used herein, a "prodrug" of a compound of the invention is a functional derivative of the compound which, upon administration to a patient, eventually liberates the compound of the invention in vivo. Administration of a compound of the invention as a prodrug may enable the skilled artisan to do one or more of the following: (a) modify the onset of the compound in vivo; (b) modify the duration of action of the compound in vivo; (C) modify the transportation or distribution of the compound in vivo; (d) modify the solubility of the compound in vivo; and (e) overcome a side effect or other difficulty encountered with the compound. Typical functional derivatives used to prepare prodrugs Include modifications of the compound that are chemically or enzymaticaily cleaved in vivo. Such modifications, which Induce the preparation of phosphates, amides, esters, thioesters, carbonates, and carbamates, are well known to those skilled in the art.

The invention also provides a compound of the invention for use in medical therapy, particularly in bacterial Infections. Thus, in a further aspect, the invention Is directed to the use of a compound according to Formulas (I) to (IX), respectively, or a pharmaceutically-acceptable salt thereof in the preparation of a medicament for the treatment of bacterial infections.

COMBINATION THERAPIES

Active drug or therapeutic agents, when employed in combination with the compounds, or pharmaceutical compositions of the present invention, may be used or administered, for example, in dosage amounts Indicated in the Physicians' Desk Reference (PDR) or as otherwise determined by one of ordinary ski!l in the art.

In the context of this specification, the term "simultaneously" when referring to simultaneous administration of the relevant drugs means at exactly the same time, as would be the case, for example in embodiments where the drugs are combined in a single preparation. In other embodiments, "simultaneously" can mean one drug taken a short duration after another, wherein "a short duration" means a duration which allows the drugs to have their intended synergistic effect.

In light of the foregoing, the present invention also relates to a combination therapy, which may be a comprised of a simultaneous or co-administration, or serial administration of a combination of compounds or pharmaceutical compositions of the PU85023 present Invention with other active drug or therapeutic agents, such as described above, and where such administration also is determined by one of ordinary skill in the art.

in addition, the present invention also relates to a combination therapy for the treatment or prevention of repiratory tract or respiratory diseases as described herein, which is comprised of a composition, dosage form or formulation formed from a synergistic combination or mixture of compounds, controlled release compositions, dosage forms or formulations of the present invention and another active drug or therapeutic agent or agents as those described above and optionally which comprises pharmaceutically acceptable carrier, diluent or adjuvent. In such an aforementioned combination composition, dosage form or formuiation of the present invention, each of the active drug components are contained in therapeutically effective and synergistic dosage amounts. The Examples set forth below are illustrative of the present invention and are not intended to limit, in any way, the scope of the present invention.

PU85023

EXAMPLES

The following examples illustrate the Invention. These examples are not

Intended to limit the scope of the present invention, but rather to provide guidance to the skilled artisan to prepare and use the compounds, compositions, and methods of the present invention.

While particular embodiments of the present invention are described, the skilled artisan will appreciate that various changes and modifications can be made without departing from the spirit and scope of the invention. As stated above, compounds according to Formulas (!) to (IX), respectively, or pharmaceutically acceptable salts thereof are UDP-3-O— (R-3-hydroxydecanoyl)-N- acetylg!ucosamine deacetylase (LpxC) inhibitors, which are useful in the treatment of bacteria! infections. The biological activity of the compounds according to Formulas (I) and (IX), respectively, can be determined using suitable assays such as those measuring such Inhibition and those evaluating the ability of the compounds to Inhibit bacterial growth in vitro or in animal models of infection.

Antimicrobial Activity Assay

Whole-ceil antimicrobial activity was determined by broth microdi!ution using the Clinical and Laboratory Standards Institute (CLSI) recommended procedure, Document M7-A7, "Methods for Dilution Susceptibility Tests for Bacteria that Grow Aeroblca!ly". in some cases, broth microdi!ution MICs were determined using IsoSensitest broth (Oxoid) In place of CLSI recommended cation adjusted Mueller-Hinton broth. The compounds were tested in serial two-fold dilutions ranging from 0.064 to 64 ^ig/ml In the presence of 20 uM human apo-Transferrin (Sigma T-1 147).

Compounds were evaluated against Gram-negative organisms selected from

Escherichia coii, Pseudomonas aeruginosa, Proteus rnirabiiis, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Stenotrophomonas maltophilia,

Acinetobacter baumanii, Serratia marascens, Citrobacter freundii, and Klebsiella oxytoca.

The minimum inhibitory concentration (MIC) was determined as the lowest concentration of compound that inhibited visible growth. A mirror reader was used to assist in determining the MIC endpoint.

Each of the listed Examples 1 to 56 and 59 to 61 identified in the present PU85023 application, were tested in at least one exemplified salt or free base form. Unless otherwise noted, the tested the aforementioned examples each had a MIC≤_2,ug/ml for at least one strain of one organism listed above. Examples 5 and 47 had a MIC≤_8μ9/ΓηΙ for at least one strain of one organism listed above. Generai

Unless otherwise noted, all starting materials were obtained from commercial suppliers and used without further purification. Unless otherwise indicated, all temperatures are expressed in °C (degrees Centigrade). Unless otherwise indicated, all reactions are conducted under an Inert atmosphere at ambient temperature.

All temperatures are given in degrees Celsius, all solvents are highest available purity and ail reactions run under anhydrous conditions in an argon (Ar) or nitrogen (N₂) atmosphere where necessary.

¹H NMR (hereinafter also "NMR") spectra were recorded on Brucker AVANCE- 400 spectrometers. CDCI₃ is deuteriochloroform, D SO-D₆ Is

hexadeuteriodimethyisulfoxide, D₂0 is Deuterium oxide, and CD₃OD is

tetradeuteriomethanol. Chemical shifts are expressed In parts per million (ppm, 6 units). Coupling constants are in units of hertz (Hz), Splitting patterns describe apparent multiplicities and are designated as s (singlet), d (doublet), t (triplet), q (quartet), quint (quintet), m (multiplet), br (broad).

Mass spectra were run on open access LC-MS systems, either a PE Sciex Single

Quadrupole LC/MS API-15Q or a Waters. The compound is analyzed using a reverse phase column, e.g., Xbridge-G18, Sunfire-C18, Thermo Aquasil/Aquasi! C18, Acquity HPLC C18, Thermo Hypersil Gold eluted using an acetonitri!e and water gradient with a low percentage of an acid modifier such as 0.02% TFA.

Analytical HPLC was run using an Agilent system (1 100 series) with variable wavelength UV detection using Sunfire-C18 analytical columns and reverse phase chromatography with acetonitrile and water gradient with a 0.05 or 0.1 % TFA modifier (added to each solvent). Preparative HPLC was performed using a Giisort Preparative System with variable wavelength UV detection and Sun Fire C18 preparative columns. The compounds are eluted using a gradient of acetonitrile and water. Neutral conditions used an acetonitrile and water gradient with no additional modifier, acidic conditions used an acid modifier, usually 0.05 % or 0.1 % TFA (added to both the acetonitrile and water).

Flash chromatography was run using a Teledyne !sco Comh!f!ash RF or

Companion, with normal or reverse phase, disposable Redi-Sep flash columns, and a PU85023 detector with UV wavelength at 254 nm and a variety of solvents or solvent combinations.

Heating of reaction mixtures with microwave Irradiations was carried out on a Blotage Microwave.

Compound Examples

Examp e 1 1-(((6RJR)-7-((Z)-2-((((S)-4-(tert-buto^^

dioxobutan-2-y!)oxy)imino)-2-(2-((tert-buto

(((4-meihoxybenzy!)oxy)carbony!)~8-oxo~5-thia~1-azablcyclo[4.2.0]oct-2~en-3~yi)methyl)-1 - (2-(5-chioro-6 _>7-bis((4-methoxybenzyi)oxy)-1 -oxo-3_l4-dihydroisoquino!in-2(1 H)- y!)ethyi)pyrrolidin-1-ium

Example l a (E)-2-chloro-3,4-dimethoxy-1-(2-nitrovinyi)benzene

The mixture of 2-chloro-3,4-dimethoxybenzaidehyde (5g, 24.92 mmoi) (Aldrich) and nitromethane (6,8 mi, 122 mmoi) and ammonium acetate (1.921 g, 24.92 mmoi) was reflux in glacial acetic acid (20 mL) for 4h. Mixture was diluted with brine (300 mi) and extracted with ethyl acetate (2X150 ml), washed with water, dried over magnesium sulfate and evaporated to give (E)-2-chlo! -3,4-dimethoxy~1 -(2~nitrovinyi)benzene (8.07 g, 24.92 mmoi, 100 % yield) . LCMS (M+H)⁺ : 244.0

-(2-Chloro-3,4-dimethoxyphenyi)ethanamine

To tetrahydrofuran (THF) (60 mL) was added lithium aluminum hydride (1.822 g,

48.0 mmoi) at 0 °C and aluminium chloride (6.40 g, 48.0 mmoi) In tetrahydrofuran (THF) PU85023

(80 mL) was added via syringe . The solution of (E)-2-chioro-3,4-dimethoxy-1-(2- nitrovinyi)benzene (5.85 g, 24 mmoi) in tetrahydrofuran (THF) (100 mL) was added via addition funnel over 40 min. Stirred at rt for 16h. Water (8ml) was added dropwise followed by 5N hydrochloric acid (60 ml). The mixture was washed with ether (2X200mL) and the aqueous phase was made basic with 6N sodium hydroxide. The aqueous layer was extracted with ether (3X200mL). Dried over magnesium sulfate, evaporated to give 2-(2-ch!oro-3,4-dimethoxyphenyl)ethanamine (2.85g, 13.21 mmoi, 55.1 % yield) .LC S (M+Hf : 216.0

Exarnpfe 1 c Ethyl 2-chloro-3,4-dimethoxyphenethylcarbamate

To the solution of 2~(2-chioro-3,4-dimethQxyphenyl)ethanamine (2,8g, 12.98 mmoi) In dichioromethane (DCM) (40 mL) at 0 "C was added triethylamine (1.990 mL, 14.28 mmoi) and ethyl chloroformate (1 .371 mL, 14.28 mmoi). The reaction mixture was stirred at room temperature for 12 hours. The resulting mixture was washed with water, brine, dried over magnesium sulfate and chromatographed to give ethyl 2-chloro-3,4- dimethoxyphenethylcarbamate (1 .7g, 5.91 mmoi, 97 % yield). LCMS (M+H)⁺ : 288.1 Exarnpfe 1 d 5-Chioro-6,7-dimethoxy-3,4-dihydroisoquinolin-1 (2H)-one

To ethyl 2-chloro-3,4-dimethoxyphenethylcarbamate 3 (2g, 6.95 mmoi) in 1 , 1 , 1 ,3,3,3-hexamethyidisiioxane (7.39 ml, 34.8 mmo!) in a 20 ml microwave tube was added phosphorous oxichloride (6,48 ml, 69.5 mmoi) and phosphorus pentoxlde (4.93 g, 34,8 mmoi). Microwaved at 150 °C (high absorption setting) for 1 hour. Solvent removed and poured on to the ice. Neutralized with 2M sodium hydroxide and extracted with dichioromethane. Dried over magnesium sulfate and chromotagraphed eiuting with 0- 20% methanokdich!oromethane to give 5-chloro-6 J-dimethoxy-3,4-dihydroisoquinolin- 1 (2H)-one (340 mg, 1 .407 mmoi, 20.24 % yield) . ) LCMS (M+H)⁺ : 242.1

Exarnpfe 1e 5-Chloro-6,7-dihydroxy-3,4-dihydroisoquinolin-1 (2H)-one PU85023

To the solution of 5-chioro-6,7-dimethoxy-3,4-dihydroisoquinolin-1 (2H)-one (700 mg, 2.90 mmol) in dichioromethane (DC ) (2 mL) at -78 °C was added borontribromide (0.821 mL, 8.69 mmoi). Mixture was allowed to warm up to room temperature and stirred at room temperature for 1 hour. Mixture was diluted with methanol and stripped several times to give 5-chioro-6,7-dihydroxy-3,4-dihydroisoquinoiin-1 (2H)-one (619 mg, 2.90 mmol, 100 % yield) LCMS (M+H)⁺ : 214.0

Example 1f 5-Chloro-6,7-bis((4-methoxybenzyl)oxy)-3,4-dihydroisoquinoiin-1 (2H)-one

To the solution of 5~chloro~6,7~rJihydroxy~3,4~dihydroisQquinoiin-1 (2H)-one (0.619 g, 2.9 mmol) in N,N-dimethylforrnamide (DMF) (10 mL) was added potassium carbonate (1.202 g, 8.70 mmol) followed by 1-(chioromethyl)-4-methoxybenzene (0.767 mL, 5.66 mmol). Stirred at room temperature for 4hours. The mixture was extracted with ethyl acetate, washed with water, sodium chloride, and dried over magnesium sulfate. The crude mixture was chromotagraphed on silica ISCO column to give 5-chioro-6J-bis((4- methoxybenzyl)oxy)-3.4-dihydroisoquinolin-1 (2H)-one (1.017 g, 2.241 mmol, 77 % yle!d). LCMS (M+H)⁺: 454.4 Exarnpfe 1 g 5-Chloro-6,7-bis((4-methoxybenzyl)oxy)-2-(2-(pyrrolidin-1-yl)ethyl)-3,4- dihydroisoquinolin-1 (2H)-one

To the solution of 5-chioro-6₁7-bis((4-methoxybenzy!)oxy)-3,4-dihydroisoquinolin- 1 (2H)-one (1.017g, 2.241 mmoi) in Ν,Ν-dimethyiformamide (DMF) (10 ml) was added sodium hydride (0.179 g, 4.48 mmol) .Stirred at rt for 10 min. 1-(2-Chloroethyl)pyrrolidine (0.329 g, 2.465 mmol) (first freed as free base with MP-carbonate resin) was added and the mixture was heated at 40 °C for 12h. Mixture extracted with EtOAc, washed with water, NaC!, dried over magnesium sulfate. Chromotagraphed eluting with 0-10% methanolidichioromethane to give 5-chloro-6,7-bis((4-methoxybenzyl)oxy)-2-(2- PU85023

(pyrrolidin-1 -yl)ethyl)-3,4-dihydroisoquinolin-1 (2H)-one (420 mg, 0.782 mmoi, 34.0 yield) . LCMS (M+H)⁺: 551.0

Example 1 (S)-4-tert-buty! 1-(4-methoxybenzyi) 2-(((Z)-(1-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)-2-(((5R,6R_I7R)-3-(iodometr!yl)-2-(((4- methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-en-7- y!)arnino)-2-oxoethylidene)amino)oxy)succinate

To the solution of (S)~4-tert~butyi 1-(4-methoxybenzyl) 2-(((Z)-(1-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)-2-(((5R,6R,7R)-3-(chloromethyl)-2-(((4- methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1 -azabicycio[4,2.0]oct-2-en-7- y!)amino)-2-oxoethylidene)amino)oxy)succinate (1.183 g, 1.25mmoi) in acetone (50 mL) at room temperature was added sodium iodide (0.234 g, 1 .563 mmoi). The mixture stirred at room temperature for 2hours. Fiitered and evaporated to give (S)-4-tert-butyl 1-(4- methoxybenzyl) 2-(((Z)-(1-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)-2-(((5R,6R,7R)-3- (iodomethyi)-2-(((4-methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-en-7-yl)amino)-2-oxoethyiidene)amino)oxy)succinate (1 .1 g, 1.080 mmoi, 85 % yield). LCMS (M+H)⁺: 1038.9

Exampje 1 i 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl)oxy)-1 ,4- dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetami (((4-methoxybenzyl)oxy)carbony!)-8-oxo-5-thia-1-azabicycio[4.2 ]od-2-en-3-yl)methyi)-1 - (2-(5-chloro-6,7-bis((4-methoxybenzyi)oxy)-1 -oxo-3,4-dihydroisoquinolin-2(1 H)- yi)eihy!)pyrrolidln-1~lum PU85023

To the heat gun dried flask under nitrogen added 5-chioro-6,7-bis((4- methoxybenzyl)oxy)-2-(2-(pyrrolidin-1 -yi)ethyl)-3,4-dihydroisoquinolin-1 (2H)-one 7 (300 mg, 0.544 mmol) in Ν,Ν-dimethyiacetamide (DMA) (10 mL) followed by (S)-4-tert-butyl 1- (4-methoxybenzyl) 2-(((Z)-(1 -(2-((tert-butoxycarbonyl)amino)thiazol-4-yl)-2-(((5R,6RJR)- 3-(iodomethyl)-2-(((4-met oxybenzyl)oxy)carbonyi)-5-oxido-8-oxo-5-triia-1- azabicyclo[4.2.0]oct-2-en-7-yl)amino)-2-oxoethylidene)amino)oxy)succinate (585 mg, 0.544 mmol) in Ν,Ν-dimethylacetamide (DMA) (10 mL) . The mixture was heated at 40 °C for 3hours, and was then cooled and stored in refrigerator overnight. The mixture was cooled to -40 °C and Ν,Ν-dimethylformamide (DMF) (10 mL) was added followed by phosphorus tribromide (0.103 mL, 1 .089 mmol) dropwise over 10 min. Srirred at -40 °C for 30 minutes. Poured Into the cooled solution of 5% sodium chloride. The resulting solid was filtered and dried to give crude 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4- methoxybenzyl)oxy)-1 ,4-dioxobutan-2-yl)oxy)imino)-2-(2-((tert- butoxycarbonyl)amino)thiazol-4-yl)acetamido)-2-(((4-methoxybenzyl)oxy)carbonyi)-8-oxo- 5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-(2-(5-chloro-6,7-bis((4- methoxybenzyl)oxy)-1 -oxo-3,4-dihydroisoquinolin-2(1 H)-yl)ethyl)pyrroiidin-1-ium (120 mg, 0.083 mmol, 15.24 % yield) . LCMS: (M+Hf: 1447.5 Example 1 j 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl)oxy)-1 ,4- dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetam

(((4-methoxybenzyl) oxy)carbonyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)- 1-(2-(5-chloro-6,7-bis((4-met oxybenzyl)oxy)-1 -oxo-3,4-dihydroisoquinolin-2(1 H)- y!)ethyl)pyrrolldln~1-lum

To the solution of 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-bLitoxy)-1-((4- methoxybenzyl)oxy)-1 ,4-dioxobutan-2-yi)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)t iazoi-4-yl)acetamido)-2-(((4-methoxybenzyl)oxy)carbonyi)-8-oxo- 5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-(2-(5-chioro-6,7-bis((4- methoxybenzyl)oxy)-1-oxo-3,4-dihydroisoquinolin-2(1 H)-yl)ethyl)pyrrolidin-1-ium (170 mg, 0, 1 18 mmol) in dichlorornethane (DCM) (2 mL) was added anisole (0.385 mL, 3.53 mmoi) followed by trifluroacetic acid (0.543 mL, 7.05 mmoi). Stirred at room temperature for 24 hours. Solvent removed and the mixture triturated with isopropyi ether. The resulting solid was filtered. The crude solid was dissolved in acetonith!e, water, hydrochloric acid.

HP20ss resin was added and the mixture was concentrated to dryness. The compound absorbed on to the resin and passed through plug of HP20ss on C18 ISCO column. Purified by Gilson HPLC (e!uting with acetonitrl!e :water 0.1 % trifluroacetic acid}. The acetonitrile evaporated. Resulting aqueous solution was basified to PH 6.0 with 0.2 sodium hydroxide. Lyophiilzed to give 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)- 1 ,2-dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2- en-3-yl)methyi)-1-(2-(5-chloro-6,7-dihydroxy-1-oxo-3,4-dihydroisoquinoiin-2(1 H)- y!)ethyi)pyrrolidln-1-ium, 2 Sodium salt (14 mg, 0.016 mmol, 13.37 % yield). LCMS (M+H)⁺: 809. 5.

Ή N R (400 MHz, DEUTERIUM OXIDE) 5ppm 1.77 - 2.02 (m,6 H) 2.12 (d, J=5.81 Hz, 4H) 2.48 - 2.73 (m, 3 H) 2.87 (br. s., 3 H)) 3.81 (br. s., 4 H) 3.92 - 4.03 (m, 2 H) 4.04 (br. s., 2 H) 4.78 - 4.90 (m, 3 H) 5.24 (d, J=4.55 Hz, 2 H) 5.73 (d, J=4.55 Hz, 2 H) 6.90 (s, 2 H) 7.18 (br. s., 1 H) Example 2 (6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-(2-(5-hydroxy-1-methyl-4-oxo-1 ,4-dihydropyridine-2-carboxamido) ethyl)pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2- carboxyiate, 3 Sodium salt PU85023

E amp e 2a 2-(hydroxymethyl)-5-((4-methoxybenzyi)oxy)-4H-pyran-4-one

To a solution of 5-hydroxy-2-(hydroxymethyl)-4H-pyran-4-one (13.35 g, 94 mmol) In Ν,Ν-dimethylformamide (180 mi) stirred under nitrogen at 0°C was added a solution of potassium tert-butoxide (12.65 g, 1 13 mmoi) in Ν,Ν-dimethylformarnide (40.0 ml) dropwise over 10 min. The mixture was then stirred at room temperature for 90 minute and 4-methoxybenzy! chloride (15.5 ml, 1 14 mmol) was added dropwise over 15 min and allowed to stir at 50 °C for 3 days. The reaction was cooled and concentrated under vacuum. Water (400 mL) was added, the resulting precipitate was filtered, washed with water, collected and dried to afford 2-(hydroxymethyl)-5-((4-methoxybenzy!)oxy)-4H- pyran-4-one (18.17 g, 69.3 mmol, 73.8 % yield) as tan solid. LCMS: ( +Hf : 263.0

E am le 2b 5-((4-methoxybenzyl)oxy)-4-oxo-4H-pyran-2-carboxylic acid

To a solution of 2-(hydroxymethyl)-5-((4-methoxybenzyl)oxy)-4H-pyran-4-one (2.57 g, 9.80 mmoi) in chloroform (45 ml) was added dimethyl sulfoxide (12 ml) and triethylamine (8.20 mi, 58.8 mmoi). The mixture was cooled to 0 °C, and sulfur trioxide, PU85023 pyridine salt (3.96 ml, 49.0 mmol) was added. The mixture was allowed to warm to room temperature and stirred for 20 h. The resulting precipitate was filtered and washed with water. The fiitrate was washed with water and the phases were separated. The organic layer was concentrated and combined with the precipitate. Water (20.0 ml), acetone (20.0 ml) and sulfamic acid (1 .431 g, 14.74 mmol) were added, followed by sodium chlorite (1 .332 g, 14.73 mmol) and the mixture was allowed to stir at room temperature for 70 h. The resulting precipitate was filtered, washed with water and acetone, and dried to give, 5-((4-methoxybenzyi)oxy)-4-oxo-4H-pyran-2-carboxylic acid (1 .466 g, 5.31 mmol, 54.2 % yield) as white solid. LG S: ( +Naf: 299.0.

Exarnpfe 2c 5-((4-methoxybenzyl)oxy)-1-methyl-4-oxo-1 ,4-dihydropyridine-2-carboxylic acid

A mixture of 5-((4-methoxybenzy!)oxy)-4-oxo-4H-pyran-2-carboxylic acid (1 .4627 g, 5.30 mmo!) and methanamine (1 1.0 ml, 22.00 mmol) was stirred at room temperature for 20 h, The reaction was concentrated under reduced pressure, and the residue was dissolved in 15 mi_ of de-ionized water. The aqueous solution was cooled in an ice bath and treated dropwise with 2N hydrochloric acid (3.0 ml, 6.00 mmol). The resulting yellow solid was filtered, washed with water and acetone, and dried to give 5-((4- methoxybenzyl)oxy)-1-methyl-4-oxo-1 ,4-dihydropyridine-2-carboxylic acid (1 .21 g, 4.18 mmol, 79 % yield). LCMS: (M+H)⁺: 290.1.

Example 2d 5-((4-methoxybenzyl)oxy)-1-methyl-4-oxo-N-(2-(pyrrolidin-1-yl)ethyl)-1 ,4- dihydropyridine-2-carboxamide

PU85023

A mixture of 5-((4~FTiethoxybenzy!)oxy)-1 ~FTiethy!-4-oxo~1 ,4-"dihydropyridine-2- carboxyiic acid (1.21 g, 4.18 mmol), HATU (1.908 g, 5.02 mmoi),diisopropy!ethy! amine (1.5ml, 8.59 mmol) in N,N-dimethylformarnide(15.G mi) was stirred at room temperature for 30min. at which time 2-(pyrro!idin-1-yl)ethanamine (0.58 ml, 4.61 mmol) was added and the reaction was allowed to stir for 3 h. The reaction was concentrated under vacuum, the residue was triturated with dich!oromethane. The solid materia! was filtered and further washed with d!ehioromethane. The supernatant was collected, concentrated under reduced pressure and the residue was purified on silica gel eluting with methanoi/dich!oromethane/ammonium hydroxide. The fractions containing the desired product were concentrated, the residue was dissolved in dieh!oromethane, and washed with water. The organic phase was dried with anhydrous sodium sulfate, filtered, and concentrated to give 5-((4-methoxybenzy!)oxy)-1-methyl-4-oxo-N-(2-(pyrrolidin-1- y!)ethyl)-1 ,4-dlhydropyridine-2-carboxamide (1.32 g, 3.42 mmol, 82 % yield) as nearly white solid. LC S: ( +H)⁺: 388.2

Example 2e 1-(((6R,7R)~7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4~methoxybenzyl)oxy)-1 ,4- dioxobutan-2-y!)oxy)lmino)~2-(2~((tert-buto^

(((4-methoxybenzyl)Qxy)carbonyi)-8~oxo~5-thia~1-azablcyclQ[4.2.0]oct-2~en-3-yi)methyl)-1 - (2-(5-((4-methoxybenzyl)Qxy)-1 -methyl-4-oxo-1 ,4-dlhydropyrldine-2- carboxamido)ethyl)pyriOlidin~1-ium

PU85023

To the solution of (S)-4-tert-butyl 1-(4-methoxybenzyl) 2-(((Z)-(1-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)-2-(((5R,6R,7R)-3-(iodomethyl)-2-(((4- methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-en-7- yl)amino)-2-oxoethylidene)amino)oxy)succinate (Example I n) (900 mg, 0.887 mmol} in Ν,Ν-dimethyiacetamide (DMA) (20 mL) was added 5-((4-methoxybenzyl)oxy)-1 -methyl-4- oxo-N-(2-(pyrrolidin-1-yi)ethyl)-1 ,4-dihydropyridine-2-carboxamide { (334 mg, 0.867 mmol). The mixture heated at 40 °C for 3 h, and was then cooiled and stored in fridge overnight. The mixture was cooled to -40 °C and N,N-dimethylforrnamide (DMF) (20 mL) was added, followed by phosphorus tribromide (0.103 mL, 1.089 mmol) dropwise over 10 rnin. Stirred at -40 °C for 30 minutes. Poured into the colled solution of 5% sodium chloride. The resulting solid filtered and dried to give crude 1-(((6R,7R)-7-((Z)-2-((((S)-4- (tert-butoxy)-1-((4-methoxybenzyi)oxy)-1 ,4~dioxobutan-2~yi)oxy)imino)-2~(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)acetamido)-2-(((4-methoxybenzy!)oxy)carbonyi)-8-oxo- 5-thia-1-azabicyclo[4.2 ]od-2-en-3-yl)methyl)-1 -(2-(5-((4-methoxybenzyi)oxy)-1-methyi- 4-oxo-1.4-dihydropyridine-2-carboxamido)ethyl)pyrrolidin-1-iLim (1 1 10 mg, 0.867 mmol, 100 % yield). Used in the next step without further purification. LCMS (M+H)⁺ : 1281 .8

Exampte 2f (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyeihoxy)im!no) acetamido)-3-((1-(2~(5-hydroxy-1-methyl-4~oxo-1 ,4-dlhydropyrid!ne-2-carboxamldo) ethyi)pyrrolidln-1-!um-1-yi)methy!)-8-oxo-5-thia-1 -azabicyclo[4.2.0joci~2-ene-2- carboxylate, 3 Sodium salt

PU85023

To the soiution of 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl) oxy)-1 ,4-dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazoi-4- y!)acetamido)-2-(((4-methoxybenzyi)oxy)carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- en-3-yl)methyi)-1 -(2-(5-((4-methoxybenzyl)oxy)-1-methyl-4-oxo-1.4-dihydropyridine-2- carboxamido)ethyl)pyrrolidin-1-ium (1.1g, 0.859 mmol) in dich!oromethane (DCM) (20 ml) at 0 °C was added anisoie (0.938 mL, 8.59 mmol) and tnf!oroacetic acid (1.324 mL, 17.18 mmol). The mixture was stirred at room temperature overnight. The solid was triturated with isopropyl ether and dissolved with acetonitrlle, water, 2N hydrochloric acid. Compound was loaded onto HP20 resin and pased trough C18 ISCO column.

Evaporated and the resulting aqueous solution was basified to PH 8.0 with 0.2M sodium hydroxide. Lyophiiized to give (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-3-((1 -(2-(5-hydroxy-1-methy!-4-oxo-1 ,4- dihydropyridine-2-carboxamido)ethyl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 - azabicyclo[4.2.0]oct-2-ene-2-carboxylate, 3 Sodium salt (40 mg, 0.048 mmol, 5.60 % yield). , LCMS( M+H)⁴ : 763.3

'H N R (400 MHz, DEUTERIUM OXIDE) δ ppm 1.91 - 2.04 (m, 1 H) 2.14 (d, J=2.78 Hz, 3 H) 2.52 - 2.86 (m, 2 H) 2.80 (s, 1 H) 2.81 - 2.92 (m, 1 H) 2.96 (s, 1 H) 3.23 - 3.61 (m, 1 1 H) 3.61 - 3.78 (m, 4 H) 3.61 - 3.68 (m, 5 H) 3.75 - 3.90 (m, 2 H) 4.06 (d, J=13.89 Hz, 1 H) 4.79 - 4.92 (m, 1 H) 5.25 (d, J-4.55 Hz, 1 H) 5.72 (d, J-4.55 Hz, 1 H) 6.57 (s, 1 H) 6.90 (s, 1 H) 7.47 (s, 1 H)

Example 3 (6R,7R)~7-((Z)-2-(2~aminothlazol~4-yl)~2-(((S)-1 ,2-dicarboxyethoxy)!mino) acetamido)-3-((1 -(2-(7-hydroxy-1 _!8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- y!)ethyl)pyrrolidln-1 -lum-1-yl)methy!)~8-oxo-5 hia-1-azablcyclo[4.2.0]oct~2-ene-2- carboxyiate

Example 3a 7-((4-methoxybenzy -pyrido[1 ,2-a]pyrazme-1 ,8(2H)-dioRe

The mixture of 5-((4-methoxybenzyl)oxy)-4-oxo-4H-pyran-2-carboxyiic acid

(Example 2b) (5.24 g, 18.21 mmo!), ethane-1 ,2-diamine (1.830 mL, 27.3 mmol) in water (80 mL) was stirred at 80 ^t!C for 1 h and then 80 °C for 20 h. Yeilow soiid precipitated from the reaction when heating. After cooling, white precipitate was filtered off, washed with 20 mL of cold water/acetone (v:v = 1 :1 ) and dried to give 7-((4-methoxybenzy!)oxy)-3,4- dihydro-1 H-pyridoi1 ,2-a]pyrazine-1 ,8(2H)-dione (4.15 g, 13.82 mmol, 76 % yield) as white solid, which was used in the next step without further purification. LCMS( M+H)⁺: 301 .2

Exampje 3b 7-((4-methoxybenzyl)oxy)-2-(2-(pyrro!idin-1-yi)ethyl)-3,4-dihydro-1 H- pyrido[1 ,2-a]pyrazine-1 ,8(2H)-dione

To a solution of 7-((4-methoxybenzyi)oxy)-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazine- 1 ,8(2H)-dione (4.15 g, 13.82 mmoi) in Ν,Ν-dimethylformamide (DMF) (50 mL) was added sodium hydride (1.382 g, 34.5 mmoi) at 0 °C, and the mixture was stirred at room temperature for 30 min. Then the mixture of 1~(2-chloroethyl)pyrro!idine, hydrochloride (Aldrich) (2.90 g, 17.03 mmoi) and triethy!amine (2.438 mL, 17,49 mmoi) in N,N- dimethylformamide (DMF) (25.00 mL) was added. The reaction was stirred at 50 °C for 30 h. The solvent was evaporated and the residue was dissolved in water (50 mL) and extracted with dichioromethane (50 mL) three times. The organic phase was dried over anhydrous sodium sulfate, filtered and evaporated to afford light yellow crude product, which was then purified by normal phase automatic silica column chromatography

(Combifiash RF), eluting with dichloromethane/methanol/ammonium hydroxide (v:v:v = 80:20:2) over 16 min to afford 7-((4-methoxybenzyl)oxy)-2-(2-(pyrrolidin-1 -yi)ethyl)-3,4- dihydro-1 H-pyridof1 ,2-a]pyrazine-1 ,8(2H)-dione (3 g, 7.55 mmoi, 54.8 % yield) as !ight yel!ow solid. LCMSi M+H)^÷: 398.0.

Example 3c 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl)oxy)-1 ,4- dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)aceta

(((4-methoxy benzyl )oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yl)met yi)-1 - (2-(7-((4-methoxybenzyi)oxy)-1 ,8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- y!)ethyi)pyrrolidln-1-ium, Iodide

To the heat gun dried flask under nitrogen was added 7-((4-methoxybenzyl)oxy)- 2-(2-(pyrrolidln-1-yl)ethyi)-3,4-dlhydro-1 H-pyrido[1 ,2-a]pyrazine-1 ,8(2H)-dione (700 mg, 1.761 mmoi) in Ν,Ν-dimethylacetamide (DMA) (10 mL) followed by (S)-4-tert-butyl 1-(4- methoxybenzyi) 2-(((Z)-(1-(2-((tert-butoxycarbonyl)amino)thiazol-4-yl)-2-(((5R,6R,7R)-3-

(iodomethyi)-2-(((4-methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-en-7-yl)amino)-2-oxoethylidene)amino)oxy)sLiccinate (Example 1 h) (1828 mg, 1 .761 mmoi) in Ν,Ν-dimethylacetamide (DMA) (10 mL) . Mixture was heated at

8 :i6 PU85023

40 "C for 3h. Cooled and stored in fridge overnight. The mixture was cooled to -40 °C and Ν,Ν-dimethy!formamide (D F) (10 mL) was added followed by phosphorus trihromide (0.332 mL, 3.52 mmol) dropwlse over 10 min. Stirred at -40 °C for 30 min. Poured Into the cooled solution of 5% sodium chloride. The resulting solid was filtered and dried to give crude 1 -(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1 -((4-methoxybenzy!)oxy)-1 ,4- dioxobutan-2-y!)oxy)imino)-2-(2-((tert-buto

(((4-meihoxybenzy!)oxy)carbony!)~8-oxo~5-thia-1-azablcyclo[4.2.0]oct-2-en-3~yi)methyl)-1 - (2-(7-((4-methoxybenzyl)oxy)-1 ,8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- y!)ethyl)pyrrolidin-1 -ium, iodide (2.8g, 1 .832 mmol, 104 % yield). LC S: (M+H)⁺: 1420.8

Exampfe 3d (6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-3-((1-(2-(7-hydroxy-1 ,8-dioxo-3,4-dihydro-1 H- pyrido[1 ,2-a]pyrazin-2(8H)-yl)ethyl)pyrrolidin-1-iLim-1 -yi)methyl)-8-oxo-5-thia-1- azabicycio[4.2.0]oct-2~ene-2-carboxylate

To a solution of 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl) oxy)-1 ,4-dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazoi-4- y!)acetamido)-2-(((4-methoxybenzyi)oxy)carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- en-3-yl)methyi)-1-(2-(7-((4-methoxybenzyi)oxy)-1 ,8-dioxo-3_>4-dihydro-1 H-pyrido[1 _l2- a]pyrazin-2(8H)-yl)ethyi)pyrro!idin-1 -ium (1.05 g, 0.812 mmol) in dich!oromethane (15 mL) stirred under nitrogen at room temperature was added neat aniso!e (0.887 mL, 8.12 mmol) in one charge. The reaction mixture was stirred for one minute and trif!uoroacetic acid (0.626 mL, 8.12 mmol) was then added. The reaction was allowed to stir at room temperature overnight and another 10eq trifluroacetic acid (0.626 mL, 8.12 mmol) was added and the reaction was allowed to stir for another 24h. Diisopropyl ether (20 mL) and water (4 mL) were added and the suspension was allowed to stir for 5 min, then decanted off the supernate from the solid mass. The solid was dissolved in acetonitrlle (10 mL), water (10 mL) and 2N hydrochloric acid (2.5 mL), absorbed onto HP20SS resin, and purified by dual HP20SS plug column and 26g C18 column eluting with 0-20% acetonitrile in water. The fractions containing pure desired compound were collected and lyophilized to give (6R,7R)-7-((Z)-2-(2- aminothiazo!-4-yl)-2-(((S)-1 ,2- PU85023 dicarboxyethoxy)imino)acetamido)-3-((1-(2-(7-hydroxy-1 ,8-dioxo-3,4-dihydro-1 H- pyrido[1 ,2-a]pyrazin-2(8H)-yl)ethyl)pyrrolidin-1-ium-1 -yi)methyl)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate (138 mg, 0.178 mmol, 21.92 % yield) as an off white solid. LCMS: (M+H)⁺: 775.4. H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 2.13 (br. s., 4 H) 2.54 - 2.71 (m, 2 H) 3.54 (br. s., 8 H) 3.74 - 3.87 (m, 8 H) 4.04 (br. s., 2 H) 4.27 (br. s., 2 H) 4.85 (br. s., 1 H) 5.23 (d, J=4.80 Hz, 1 H) 5.70 (d, J=4.80 Hz, 1 H) 6.90 (s, 1 H) 7.1 (s, 1 H) 7.57 (s, 1 H).

Example 4 (6R7 no) acetamido)-3-((1 -(2- yl)ethyl)pyrrolidin-1 -i

carboxylate, 2 Sodiu

The impure fractions of (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-3-((1-(2-(7-hydroxy-1 ,8-dioxo-3,4-dihydro-1 H- pyrido[1 ,2-a]pyrazin-2(8H)-yl)ethyl)pyrrolidin-1-ium-1 -yl)methyl)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxyiate (Example 3) were absorbed onto HP20ss resin and purified using an HP20ss-C18 plug column followed by a 26g C-18 column eluting with 0-20% acetonitriie in water. The fractions containing pure desired product were neutralized to pH 8 with 0.2 N sodium hydroxide. Quickly added a small cube of dry ice then frozen, !yophiiized to give the desired product. Isolated (S)-2-(((Z)-(1 -(2- aminothiazol-4-yl)-2-(((6R,7R)-2-carboxylato-3-((1-(2-(7-hydroxy-1 _>8-dioxo-3,4-dihydro- 1 H-pyrido[1 ,2-a]pyrazin-2(8H)-yl)ethyl)pyrrolidin-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1- azabicyclo[4.2 ]oct-2-en-7-yl)amino)-2-oxoethyiidene)amino)oxy)sLiccinate, 2 Sodium salt (45 mg, 0.055 mmol, 6.77 % yield) as an off white solid. LCMS: (M+H)⁺: 775.4. H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 2.1 1 (br. s., 4 H) 2.60 - 2.67 (m, 2 H) 3.32 - 3.65 (m, 9 H) 3.71 - 3.90 (in, 5 H) 4.05 (d, J= 14.15 Hz, 2 H) 4.25 (br. s., 2 H) 4,84 (dd, J=8.59, 4.55 Hz, 1 H) 5.22 (d, J=4.80 Hz, 1 H) 5.69 (d, J=4.80 Hz, 1 H) 6.82 - 6,94 (m, 1 H) 7.10 (s, 1 H) 7.55 (s, 1 H).

Example 5 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-3-((1-((3-hydroxy-1-methyl-4-oxo-1 ,4-dihydroquinolin- PU85023

6-yl)methy!)pyrrolidin-1-ium-1 -y!)methyi)-8^^

earboxy!ate, 3 Sodium sail

Example 5a Ethyl 4-((2-(ethoxycarbonyi)-3-oxobut-1-en-1 -yl)amino)benzoate

To a solution of (ZZE)-ethyl 2-(ethoxymethyiene)-3-oxobutanoate (4.95 g, 26.6 mmol) (commercially available from AK Scientific) in Isopropanoi (50 ml) was added ethyl 4-aminobenzoate (4.39 g, 28.6 mmol). The mixture was stirred at room temperature for 10 min, then heated to 65°C and stirred for another 5 min. The reaction was allowed to cool and was then diluted with hexane. The suspension filtered, and the precipitate was washed with hexane, and dried to give ethyl 4-((2-(ethoxycarbonyi)-3-oxobut-1 -en-1- y!)amino)benzoate (6.84 g, 22.40 mmol, 84 % yield. LCMS: ( +H)⁺: 306.0.

Example 5b Ethyl 3-acetyl-4-oxo-1 ,4-dihydroquinoline-6-carboxylate

Diphenyl ether (150 mi, 943 mmoi) was heated to boiling and ethyl 4-((2-

(ethoxycarbonyl)-3-oxobut-1-en-1-yl)amino)benzoate (6.84 g, 22.40 mmol) was added siowiy. After the addition, the reaction was allowed to stir at 200-250 °C for another 30 min. After cooling, the reaction was diluted with hexane, filtered, and the precipitate was washed with hexane/diethyi ether, collected and dried to give ethyl 3-acetyl-4-oxo-1 ,4- dihydroqulnoline-6-carboxyiate (5.74 g, 22.14 mmol, 99 % yield) as tan solid. LCMS: PU85023

(M+H)⁺: 260.0.

Exampte 5c Ethyl 3-acetyl-1-methyi-4-oxo-1 ,4-dihydroquinoiine-6-carboxyiate

To a suspension of ethyl 3-acetyi-4-oxo-1 ,4-dihydroquinoline-6-carboxylate (5.74 g, 22.14 mmoi) in Ν,Ν-dimethyiformamide (120 ml) was added potassium carbonate (9.18 g, 66.4 mmoi) and methyl iodide (4.2 ml, 67.2 mmo!). The mixture was stirred at 90

°C for 1 h. After cooling, the reaction was filtered, concentrated under high vacuum, diluted with dichioromethane/ethyl acetate, and filtered through a silica plug to afford ethyl 3-acetyl-1-methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxylate (3.64 g, 13.32 mmoi, 60.2 % yield. LCMS: (M+H)⁺: 274.0.

Example 5d Ethyl 3-hydroxy-1 -methyl-4-oxo-1 ,4-dihydroquinoiine-6-carboxylate

To a solution of ethyl 3-acetyi-1 -methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxylate (1.79 g, 6.55 mmoi) in dichioromethane(40 mi) was added meta-chioroperbenzoic acid (2.2057 g, 9.84 mmoi). The mixture was stirred at room temperature for 1 h, then concentrated under reduced pressure to roughly one half of the reaction volume. The resulting precipitate (O-acetate) was collected, added to methanol (20.0 ml), and heated at 65 °C overnight. After cooling, the reaction was filtered, and the precipitate was washed with dichloromethane to give ethyl 3-hydroxy-1-methyi-4-oxo-1.4- dihydroquinoiine-6-carboxylate (0.9956 g, 4.03 mmoi, 61 .5 % yield) as white solid.

LCMS: (M+Hf: 248.0.

Example 5e 3-((4-methoxybenzyl)oxy)-1-methyl-4-oxo-1 ,4-dihydroqiiinoiine-6- carboxyilc acid

To a suspension of ethyl 3-hydroxy-1-methyi-4-oxo-1 ,4-dihydroquinoline-6- PU85023 carboxylate (6.7 g, 27.1 mmol) in N.N-dimethylformamide (18.0 ml) at 0 °C was added sodium hydride (2.168 g, 54.2 mmol), and the mixture was stirred for 20 min. At this time, 1-(chioromethyi)-4-methoxybenzene (3.7 ml, 27.3 mmol) was added and the reaction was heated to 75 °G and allowed to stir for 2 h. After cooling, the reaction was poured Into ice, and the resulting precipitate was filtered, washed with water and diethyl ether, collected, and dried. Methanol (24.00 mi), water (8.00 ml) and lithium hydroxide (0.86 g,

35.9 mmol) were added, and the reaction was stirred at 80 °C for 2 h. The reaction was cooled and the pH was adjusted to 3-4 by addition of 2N hydrochloric acid (18.00 ml, 36 mmol). The resulting precipitate was filtered, washed with water and diethyl ether, and dried to give 3-((4-methoxybenzy!)oxy)-1 -methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxylic acid (8.1937 g, 18.25 mmol, 67.4 % yield, LCMS: (M+H)⁺: 340.1.

Example 5f 6-(Hydroxymethy!)-3-((4-methoxybenzyl)oxy)-1-methylquinolin-4(1 H)-one

To a suspension of 3-((4-methoxybenzyl)oxy)-1 -methyl-4-oxo-1 ,4- dihydroquinoiine-6-carboxylic acid (2.7 g, 7.96 mmol) in tetrahydrofuran (THF) (100 mL) was added tr!ethy!am!ne (3.88 mL, 27.8 mmol) and ethyl chloroformate (2.369 mL, 24.67 mmol). The resulting brown suspension was stirred at room temperature for 1.5 h. The crude carbonate solution was then added to a solution of sodium borohydride (3.61 g, 95 mmol) in ethano! (100 mL) at room temperature and the resulting mixture was stirred at room temperature, for 1 .5 h.

The reaction was monitored by LCMS until completion. The reaction was quenched by addition of water (20 mL) and the organic voiatl!es were removed in vacuo. The residue was then diluted with water (100 mL) and extracted with dich!oromethane (2 x 100 mL). The combined extracts were washed with water (4 x 15 mL), dried over sodium sulfate, filtered and concentrated in vacuo. The residue was purified by automatic silica gel column chromatography (Combifiash RF) eluting with methanol In

dichioromethane (0- 5%) to afford 6-(hydroxymethyi)-3-((4-methoxybenzyl)oxy)-1- methylquinolln-4(1 H)-one (1.7 g, 4.81 mmol, 60.4 % yield) as a light brown solid. LCMS: (M+H)⁺: 326.3. Exam le Sg: 3-((4-methoxy ihydroquinoline-6-carbaldehyde

To a suspension of 6-(hydroxymethyl)-3-((4-methoxybenzy!)oxy)-1-methylquinoiin- 4(1 H)-one (1 .5 g, 4.61 mmoi) in dich!oromethane (DCM) (80 mL) at room temperature was added Dess-Martin penodinane (2.347 g, 5.53 mmoi). The reaction mixture soon turned into a clear brown solution and was stirred at room temperature for 1 h. Saturated sodium bicarbonate solution was then added, and the mixture was extracted with dichioromethane. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by

automatic silica gel column chromatography (Combiflash RF) eiuting with ethyl

acetate/hexanes (0-100%) to afford 3-((4-methoxybenzyi)oxy)-1-methyi-4-oxo-1 ,4- dihydroquinoiine-6-carbaldehyde (1 .1 g, 3.40 mmoi, 73.8 % yield) as a yellow solid.

LCMS: (M+H)⁺: 324.0 Example 5 3-((4-Methoxybenzyl)oxy)-1-methyi-6-(pyrro!idin-1-yimethyl)quinolin-4(1 H)-one

A solution of pyrrolidine (0.295 mL, 3.57 mmoi) and 3-((4-methoxybenzyi)oxy)-1 - methyl-4-oxo-1 ,4-dihydroquino!ine-6-carbaldehyde (1 .05 g, 3.25 mmoi) in 1 ,2- dichioroethane (DCE) (50 mL) and 1 drop of acetic acid was treated with sodium

triacetoxyborohydride (1.032 g, 4.87 mmoi). The reaction mixture was stirred for 1.5 h, and was then concentrated under vacuum. The residue was taken in water (50 mL), and extracted with dichioromethane (2 x 50 mL). The organic layers were combined, washed with brine (50 mL), dried over sodium sulfate, and concentrated. The crude product was purified twice by automatic silica gel column chromatography (Combiflash RF), eiuting with a mixture of [dichloromethane/methano!/ammonium hydroxide(80:20:2)J/

dichiormthane(0-40%) to afford pure 3-((4-methoxybenzyl)oxy)-1 -methyl-6-(pyrroiidin-1 - y!methyl)quinolin-4(1 H)-one (1 g, 2.64 mmoi, 81 % yield) as a light yellow soiild. LCMS: (M+Hf: 379.5. PU85023

Exampte 5i 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl)oxy)-1 ,4- dioxobutan-2-yi)oxy)imino)-2-(2-((tert-butoxy

methoxybenzyl)oxy)carbonyl)-8~oxo~5 hia~i-aza

((4-methoxybenzyl)oxy)-1-methyl-4-oxo-1 ,4-dihydroquinolin-6-yl)methyi)pyrroiidin-1 -

To the heat gun dried flask under nitrogen was added 3-((4-methoxybenzyl)oxy)-1- methyl-6-(pyrroiidin-1 -ylmethyl)qLiinoiin-4(1 H)-one (530 mg, 1 .400 mmol) in N,N- dimethylacetamide (DMA) (10 mL) followed by (S)-4-tert-buty! 1-(4-methoxybenzyi) 2-(((Z)-(1- (2-((tert-butoxycarbonyl)amino)thiazol-4-yl)-2-(((5R_I6R_!7R)-3-(iodomethyi)-2-(((4- methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1-azabicycio[4.2 ]oct-2-en-7-yi)amino)-2- oxoethylidene)amino)oxy)succinate (Example 1 h) (1453 mg, 1.400 mmol) in N,N- dimethylacetamide (DMA) (10 mL) . The mixture was heated at 40 °C for 3h. Cooled and stored in the fridge overnight. The mixture was cooled to -40 "C and N,N-dimethy!formamide (DMF) (10 mL) was added, followed by phosphorus tribrornide (0.284 mL, 2.80 mmol) dropwise over 10 min. Stirred at -40 ^GC for 30 m!n. Poured into the cooled solution of 5% sodium chloride. The resulting solid filtered and dried. Chromatographed on iSCO silica gel column eluting with 0-20% methano!:dichioromethane to give 1-(((6RJR)-7-((Z)-2-((((S)-4-(tert-butoxy)- 1-((4-methoxybenzy!)oxy)-1 ,4-dioxobutan-2-yl)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)acetamido)-2-(((4-methoxybenzyl)oxy)carbonyi)-8-oxo-5- thia-1 -aza bicyc!o [4.2.0] oct-2-en-3-yi)methyl)-1-((3-((4-methoxybenzyi)oxy)-1-methyi-4-oxo- 1 ,4-dihydroquinoiin-6-yl)methyl)pyrrolidin-1 -ium (400 mg, 0.314 mmol, 22 % yield). LCMS: (M+H)⁺: 1274.8 Example 5i: (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2- clicarboxyethoxy)imino)acetamldo)-3~((1-((3-hyrJroxy-1~methyi-4~oxo-1 ,4-dihydroquinolin- 6-yl)methyl)pyrrolldin-1~lum-1-yl)methyi)-8~oxo-5-thia-1-azabicyclo[4.2.0joct-2-ene-2- carboxylate, 3 Sodium salt

To the solution of 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl) oxy)-1 ,4-dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazoi-4- yi)acetamido)-2-(((4-methoxybenzyi)oxy)carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- en-3-yl)methyi)-1 -((3-((4-methoxybenzyl)oxy)-1-methyl-4-oxo-1 ,4-dihydroquino!in-6- yi)methyl)pyrroiidin-1 -ium (400 mg, 0.314 mmo!) In Dichloromethane (DCM) (5 mL) was added anisole (0.343 mL, 3.14 mmol), followed by trifluoroacetic acid (1.210 mL, 15.71 mmol). The resulting mixture was stirred at room temperature for 18h. Resulting mixture was triturated with isopropyl ether and filtered. The solid was redisoived in acetonitrile (2mL) , water (1 mL) plus 2 drops of hydrochloric acid. Loaded on to HP20ss resin and passed through HP20ss resin column. Evaporated the acetonitrile. Resulting aqueous solution was basified to PH 6.0 with 0.2M sodium hydroxide. Lyophilized to give (6R,7R)- 7-((Z)-2-(2-aminot iazol-4-yl)-2-(((S)-1 _!2-diGarboxyethoxy)imino)acetarr!ido)-3-((1 -((3- hydroxy-1-methyi-4-oxo-1 ,4-dihydroquinolin-6-yl)methyl)pyrrolidin-1-ium-1-yl)m oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2-carboxy!ate, tri sodium salt (185 mg, 0.224 mmol, 71.4 % yield). LCMS: (M+H)⁺: 758.4

'H N R (400 MHz, DEUTERIUM OXIDE) δ ppm 0.98 - 1.04 (in, 1 H) 1.95 (br. s„ 4 H) 2.07 (d, J-5.05 Hz, 3 H) 2.53 - 2.64 (m, 3 H) 3.22 (br. s., 1 H) 3.29 - 3.40 (m, 5 H) 3.40 - 3.48 (m, 2 H) 3.42 - 3.57 (m, 3 H) 3.75 - 3.86 (m, 3 H) 3.85 - 3.95 (m, 5 H) 4.08 (d,

J=13.89 Hz, 2 H) 4.44 - 4.66 (m, 3 H) 4.79 - 4.94 (m, 2 H) 5.19 - 5.25 (m, 1 H) 5.72 (d, J=4.80 Hz, 1 H) 6.81 - 6.92 (m, 1 H) 7.74 (s, 2 H) 7.92 (s, 1 H) 8.29 (s, 1 H)

Example 8 1-(((6R,7R)-7-((Z)-2-(2-aminothiazo!-4-yl)-2-(((2-carboxypropan-2-yl)oxy) lmlnQ)acetamldo)-2-carboxy-8-oxo~5-thla-1-azabicycio[4.2.0joct-2-en-3-yl)methyl)-1-(2-(3- hyrJroxy-1 -methyi-4~Qxo-1 ,4-dihydroquinoline-6-carbQxamirJo)ethyi)pyrrolidin-1~lum PU85023

Example 6a 3-((4 in-1-yl)ethyi)-1 ,4- dihydroquinoiine-6-

To a suspension of 3-((4-methoxybenzyi)oxy)-1-methyl-4-oxo-1 ,4- dihydroquinoliRe-6-earboxyiic acid (Example 5e) (0.588 g, 1.733 mmol), 2-(pyrrolidin-1 - y!)ethanamine (0.240 mi, 1.906 mmol), EDC (0.3885 g, 2.016 mmol), HO At (0.1218 g, 0.895 mmol) in dichloromethane (16 ml) was added triethylamine (1.7 ml, 12.20 mmol), and the mixture was allowed to stir at 30 °C for 24 h. After cooling, the reaction was concentrated and the residue was purified on a silica gel column (24 g), eluting with 0- 20% methanol/dichioroethane (2 % ammonium hydroxide). The desired fractions were collected, concentrated. The solid was washed with water, filtered, and dried in vacuo to afford 3-((4-methoxybenzyi)oxy)-1-methyl-4-oxo-N-(2-(pyrrolidin-1-yl)ethyi)-1 ,4- dihydroquinoline-6-carboxamide (0.595g, 1.366 mmol, 79 % yield) as white solid. LC S: (M+H)⁺: 436.2.

Exampte 6b 6R,7R)-benzhydryi 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 -oxopropan-2-y!) oxy) imino)-2~(2-((tert-butoxycarbonyl)amino)thiazQl~4-yl)acetamido)-3-(chloromethyi)-8~ oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2-carboxylate

To a solution of (Z)-2-(((1-(tert-butoxy)-2-methyl-1-oxopropan-2-yl)oxy)imino)-2- (2-((tert-butoxycarbonyl)amino)thiazol-4-yl)acetic acid (30.5 g, 71.0 mmo!) and (8R,7R)~ benzhydry! 7-amino-3-(chloromethyl)-8-oxo-5-thia-1 -azabicydo[4.2.0]oct-2-ene-2- carboxyiate, hydrochloride (32.1 g, 71 .0 mmo!) in dichioromethane (500 mL), stirred under nitrogen at ~30 C, was added neat phenyl p osphorodichioridate (12.93 mL, 85 mmoi). The reaction mixture was cooled to -40 ⁼C and AZ-methyimorphoiine (23.42 mL, 213 mmoi) was added dropwise over 30min, and stirred for an additiona! 2 hours at -40 ^GC. The reaction was then quenched by addition of 150 mL of a 10% aq. solution of citric acid, and the phases were separated. The organic phase was concentrated and the residue was then d so!ved in 500 mL of ethyl acetate. The organic solution was washed with 100mL of a 5% aq. sodium bicarbonate, 100 mL of brine, dried over sodium sulfate, concentrated under reduced pressure, and the residue was purified on a 330 g silica gei column eluting with 0-80% ethy! acetate in hexanes to afford (6R,7R)-benzhydryl 7-((Z)- 2-(((1 -(tert-butoxy)-2-methyi-1-oxopropan-2-yl)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yi)acetamido)-3-(chloromethyl)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate (42 g, 50.8 mmoi, 71 .8 % yield), ¹H NMR (400 MHz, CHLGRQFORM-c/) δ ppm 1.42 (s, 9 H) 1.55 (s, 9 H) 1 .63 (s, 3 H) 1.66 (s, 3 H) 3.52 (d, J=18.19 Hz, 1 H) 3.66 (d, J=18.19 Hz, 1 H) 4.43 (d, J=4.29 Hz, 2 H) 5.12 (d, J=5.05 Hz, 1 H) 6.08 (dd, J=8.84, 5.05 Hz, 1 H) 6.99 (s, 1 H) 7.30 - 7.49 (m, 12 H) 8.25 (d, J-8.84 Hz, 1 H). LC S: (M+H)⁺: 826.5.

Exarnpfe 8c (6R,7R)-benzhydry! 7-((Z)-2-(((1-(tert-bLitoxy)-2-methyi-1-oxopropan-2-yl) oxy)imino)-2-(2-((tert-bLitoxycarbonyl)amino)tniazol-4-yl)acetamido)-3-(ch

oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2-carboxylate 5-oxide

To a solution of (6R,7R)-benzhydryl 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)acetamido)-3- (chloromethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate (3 g, 3.63 mmoi) in dichioromethane (DCM) ( 00 mL) at -40°C was added dropwise a solution of 3- PU85023 chiorobenzoperoxoic acid (0.976 g, 4.38 mmol) in dichloromethane 20 mL) over 1 mln. The mixture was stirred at the same temperature over 30 min. LCMS indicated completion of the reaction. Dichloromethane (60 mL) was added and the mixture was treated with 15% aqueous sodium thiosuifate(40 mL). The organic solution was separated and washed with 5% sodium bicarbonate(80 mL), brine (80 mL), dried over sodium sulfate, and concentrated. The residue was purified by ISCO automated silica gel chromatography (120 g column, 0-50% ethyl acetate/hexanes) to afford (8R,7RV benzhydry! 7-((Z)-2-(((1-(tert-butoxy)-2-methyi-1-oxopropan-2-yl)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)acetamido)-3-(chlorornethyl)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide (2.44 g, 2.90 mmoL 53.2 % yield) as a white solid. LCMS: (M+Hf: 842.2.

Example 6d (6R,7R)-benzhydryi 7-((Z)-2-(((1-(tert-butoxy)-2-met yi-1-oxopropan-2-yl) oxy)imino)-2-(2-((iert-butoxycarbonyl)am

5-thia-1-azabicyclo[4.2.0]o

To a solution of (6R,7R)-benzhydryl 7-((Z)-2-(((1-(tert-butoxy)-2-methy!-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-y!)acetamido)-3- (chloromethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate 5-oxide (2.68 g, 3.18 mmol) in acetone (40 mL) was added sodium iodide (0.715 g, 4.77 mmol). The mixture was stirred at room temperature over 3 h. LCMS indicated completion of the reaction. The solid was filtered off, the filtrate was concentrated under vacuum to afford (6R,7R)-benzhydryl 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 -oxopropan-2-yi)oxy)imino)-2-(2- ((tert-butoxycarbonyl)amino)thiazol-4-yi)acetam!do)-3-(!odomethyi)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-ox!de (2.97 g, 3.18 mmol, 100 % yield) as a dark red solid. LCMS: (M+H)⁺: 934.0.

Exampte 6e 1-(((6R,7R)-2-((benzhydryloxy)carbonyi)-7-((Z)-2-(((1-(tert-butoxy)-2- methyl-1-oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4- PU85023 y!)acetamido)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-(2-(3-((4- methoxybenzyl)oxy)-1-methyl-4~oxo-1 ,4-dihydroquinoiine-6-carboxamido)ethyl)pyrroiidin- 1-ium

To a solution of 3-((4-methoxybenzyi)oxy)-1 -methyi-4-oxo-N-(2-(pyrrolidin-1 - yl)ethyl)-1 ,4-dihydroquino!ine-6-carboxamide (Example 6a) (200 mg, 0.459 mmoi) in N,N- dimeihyiaceiamide (8 mL) stirred under nitrogen at room temperature was added solid (6RJR)-benzhydry! 7-((Z)-2-(((1-(tert-butoxy)-2-methy!-1 -oxopropan-2-yi)oxy)imino)-2-(2- ((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-3-(iodomethy!)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide (429 mg, 0.459 mmoi) in one charge, and the reaction mixture was stirred at room temperature over night. The reaction was diluted with N.N-dimethyiformamide (16.00 ml), cooled to -60 °G, and 1 .1 eq (50 uL) phossphorous tribromide was added, and the reaction was allowed to warm to -40 °C. LCMS showed little sulfoxide reduction so the reaction was again cooled to -80 °C and 3.3 eq PBr₃ (0.145 mL, 1.010 mmoi) was added, and the reaction was a!!owed to warm to -40 °C. The reaction was poured into 60 mL of a 5% sodium cloride solution at 0 °C and aiiowed to stir for approximately 15 min. The insoluble material was coliecied by filtration and the solid was placed under high vacuum on a lyophilizer. The crude 1-(((6R,7R)-2- ((benzhydry!oxy)carbonyl)-7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1-oxopropan-2- y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-8-oxo-5-th azabicyclo[4.2 ]oct-2-en-3-yi)methyl)-1-(2-(3-((4-methoxybenzyl)oxy)-1-methyi-4-oxo- 1 _l4-dihydroquinoiine-6-carboxamido)ethyi)pyrroiidin-1-ium was used without further purification, isolated approximately 800 mg of crude material of unknown purity. LCMS: (M+H)⁺: 1226.2.

Examjplejf 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl) oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-en-3-yl)methyl)-1- (2-(3-hydroxy-1 -methyl-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)ethyl)pyrrolidin-1-ium PU85023

To 1-(((6R,7R)-2-((benz ydryloxy)carbonyl)-7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetamido)-8- oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yi)methyl)-1 -(2-(3-((4-methoxy benzyl )oxy)-1- methyl-4-oxo-1 ,4-dihydroquinoiine-6-carboxamido)ethyl)pyrrolidin-1-ium (250 mg, 0.204 mmol) in dicloromethane at room temperature was added anisole (0.223 mL, 2.038 mmo!) followed by infiuoroacetic acid (0.238 mL, 3.06 mmol). The reaction was allowed to stir under nitrogen for another 24h at which time the reaction appeared to be completed. Added 10 mL diisopropy! ether and 2 mL of water and let stir at room temperature for 5 min. The solution was decanted away from the solid mass, which was then dissolved in water (5 mL), acetonitriie (5 mL), and 2N HC! (1 mL). Isopropanol (5 mL) was then added, the biphaslc mixture was separated and the aqueous phase was treated with HP-20SS resin. This suspension was concentrated under vacuum and then purified on an HP20SS plug column followed by 28g C-18 column. The fractions which contained desired product were concentrated to remove the acetonitriie and lyophiiized to give 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol- -yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)meth 1-(2-(3-hydroxy-1 -methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)ethyi)pyrrolidin-1- lum (40 mg, 0.051 mmoi, 25,03 % yield) as an off white solid. LC S: (M+H)⁺: 783.5. 'H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.45 (s, 3 H), 1.46 (s, 3 H), 2.09 (br. s., 1 H) 3.48 - 3.67 (m, 7 H) 3.87 (s, 3 H), 3.89-4.0 (m,2H), 5.23 (d, J=4.55 Hz, 1 H) 6.73 (s, 1 H) 7.32 (s, 2 H) 7.76 (d, _/=9.09 Hz, 1 H) 7.92 (s, 1 H) 8.10 - 8.17 (m, 1 H) 8.70 (br. s., 1 H) 8.79 (d, 1 .77 Hz, 1 H) 9.09 (br. s., 1 H) 9.47 (d, J-8.08 Hz, 1 H). Example 7 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl) oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yl)m

(2-(3-hydroxy-1 -methyl-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)ethyl)pyrrolidin-1-iurn, sodium salt

To 1-(((6R,7R)-2-((benz ydryloxy)carbonyl)-7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetamido)-8- oxo-5-thia-1 -azabicyclo[4.2 ]oct-2-en-3-yi)methyl)-1 -(2-(3-((4-methoxy benzyl )oxy)-1- methyl-4-oxo-1 ,4-dihydroquinoiine-6-carboxamido)ethyl)pyrrolidin-1-ium (Example 8e) (175 mg, 0.143 mmol) in DC at -30 "C was added anisoie (0.158 mL, 1 .427 mmol) followed by aluminium chloride (0.713 mL, 1.427 mmol) as a 2N solution in nitromethane. Let stir at -30 ^"C for 30 min. LC S showed disappearance of starting material and conversion to product aithrough product peak did not give a strong molecular ion. Added 10 mL of isopropyl ether and 2 mL of water and let stir at room temperature for 5 min. The solid mass was suspended in the solution mixture so decantation was not possible. The mixture was filtered and the solid product was collected and dissolved in water (5 mL), acetonit iie (5 mL) and 2N hydrochloric acid(1 mL). !sopropanol (5 mL) was then added, the biphasic mixture was separated and the aqueous phase was treated with HP- 20SS resin. This suspension was concentrated under vacuum and then purified through a layer of HP20ss followed by a 26g C-18 reversed phase column. Collected UV active peak and added NaOH (0.2N) until pH ~6 then quickly added dry ice. The solution was frozen and placed on high vacuum (lyophiiizer) while frozen to afford 1 -(((8R,7R)~7-((Z)~2- (2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5- thia-1 -azabicyclo[4.2.03oct-2-en-3-yl)methyl)-1-(2-(3-hydroxy-1-methyl-4-oxo-1 ,4- dihydroqulnoline-B-carboxamidojethy^pyrrolidin-l-ium, Sodium salt (25 mg, 0.031 mmol, 21 .72 % yield) as an off white solid of - 82% purity. LCMS: (M+H)⁺: 783.5. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.31 - 1.39 (m, 8 H) 2.14 (br. s., 1 H) 3.44 - 3.55 (m, 5 H) 3.67 - 3.72 (m, 1 H) 3.89 (br. s., 3 H) 4.07 (br. s., 2 H) 5.24 (br. s., 1 H) 5.71 (d, J=4.29 Hz, 1 H) 6.73 (br. s., 1 H) 7.38 (br. s., 1 H) 7.66 (br. s., 2 H) 8.26 (br. s., 1 H). (M+H)⁺: 783.4.

Exampfe 8 1-(((6R,7R)-7-((E)-3-(2-aminothiazoi-5-yl)-3-(((R)-1 ,2-d!carboxyethoxy) lmino)~2-oxopropyl)-2-carboxy-8-oxo-5 hia-1~azabicycio[4.2.0]oct-2-en~3-yl)methyi)~1 -((1 - ethyl-6 -dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yl)methy!)pyrrolidin-1-ium, disodium salt PU85023

Example 8a Methyl 1-ethyl-6,7-dihydroxy-4-oxo-1 ,4 iihydrocinnoline-3-carboxyiate

To a suspension of 1~ethyi-4-oxo-1 ,4-dihydiO-[1 ,3]dioxoio[4,5-g]cinnollne-3- carboxylic acid (12.6 g, 48.1 rnmoi) in dichlorornethane (DGM) (50 mL} stirred under nitrogen at -30 °C was added dropwise a solution of boron trlbromide (168 ml, 168 mmo!) In dichlorornethane (140 mL). The white suspension turned yellowish. The mixture was kept at -30 ^C'C for 1 h and then allowed to warm up to room temperature, and stirred for 40 h. The reaction mixture was again cooled to -30 °C, methanol (100 mL) was added, and the resulting mixture was stirred at room temperature over the weekend. The organic solvents were partially removed to afford a light yellow suspension. The solid was collected by filtration and washed with dichlorornethane (20 mL). The filtrate was concentrated again to provide more yellow solids that was collected and washed by dichlorornethane (20 mL). The solids were combined to afford methyl 1-ethy!-6,7- dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxylate (1 1 g, 35.8 mmol, 74.5 % yield). LCMS: (M+Hf: 265.0

E am le 8b Methyl 1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoline- 3-carboxyiate

To a suspension of methyl 1~ethyi~8,7~dihydroxy~4~oxo~1 ,4~dihydrocinnoiine-3- carboxylate (1 1 g, 35.8 mmol), potassium carbonate (16,33 g, 1 18 mmol) and potassium Iodide (1 .189 g, 7.16 mmol) in acetone (200 mL) at room temperature, was added 1- (chloromethyl)-4-methoxybenzene (16.82 g, 107 mmol). The mixture was then heated to reflux overnight. After cooling to room temperature, the mixture was concentrated in vacuo and the residue was partitioned between dichlorornethane and brine. The organic PU85023 phase was separated, and the aqueous phase was extracted with dichioromethane several times. The combined organic extracts were dried over sodium sulfate, filtered and concentrated. The residue was purified by automatic si!ica gel column chromatography (Combifiash RF) eluting with methanol/ dichioromethane (0-20%) to afford methyl 1 -ethyi- 6,7-bis((4-met oxybenzy!)oxy)-4-oxo-1 ,4-diriydrocinnoline-3-carboxylate (15 g, 24.88 mmo!, 68.9 % yield) as a light yellow solid. LC S: (M+H)⁺: 505.1.

Example 8c 1-Ethyi-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydrocinnoline-3- carboxyilc acid

To a suspension of methyl 1-et yl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 _>4- dihydrocinnoline-3-carboxylate (3 g, 5.95 mrnoi) in a mixed solvent of water (25.00 mL) and methanol (75 mL) was added potassium hydtoxide (1 .888 g, 29.7 mmo!). The mixture was stirred at room temperature for 0.5 h and then heated to reflux for 1 h when the yellow suspension turned Into a clear yellow solution. LCMS indicated completion of the reaction. The mixture was cooled to room temperature and concentrated. The residue was dissolved in water (100 mL), and was then acidified to ~pH 1 using aq. 6N hydrochloric acid. The white precipitates were collected by filtration, and the filtrate was extracted with ethyl acetate (2 x 100 mL). The combined organic layers were

concentrated to provide more white solids that were combined with previous white precipitates and dried under high vacuum to afford 1-ethyl-6,7-bis((4-methoxybenzyl)oxy)- 4-oxo-1 _>4-dihydrocinno!ine-3-carboxyiic acid (2.8 g, 5.71 mmo!, 98 % yield). LCMS: (M+Hf: 491.4. Example 8d 1-ethyl-3-(hydroxymethyl)-6,7-bis((4-methoxybenzyl)oxy)cinnolin-4(1 H)-one

To a suspension of 1-eihyl-6,7-bis((4-methoxybenzy!)oxy)-4-oxo-1 ,4- dihydrocinnoline-3-carboxyiic acid (2.25 g, 4.59 mmo!) In tetrahydrofuran (THF) (60 mL) was added trlethylamlne (2.1 10 mL, 15.14 mmo!) and ethyl chloroformate (1.322 mL, PU85023

13.78 mmol). The resulting yellow solution was stirred at room temperature for 2 h, and was then added to a solution of sodium borohydrlde (2.083 g, 55.0 mmol) in ethanoi (80 mL). After being stirred for 2 h, the reaction was quenched by addition of water (5 mL). The organic solvent was removed in vacuo, the residue was then taken in water (50 mL) and extracted with dich!oromethane (2 x 50 mL). The combined extracts were dried over sodium sulfate, filtered and concentrated in vacuo. The crude product was purified by automatic silica ge! column chromatography (Combif!ash RF), eiuting with

methanol/dich!oromethane (0- 10%) to afford 1-ethyi-3-(hydroxymethyi)-6,7-bis((4- methoxybenzyl)oxy)cinnolin-4(1 H)-one (1 .97 g, 3.84 mmol, 79 % yield) as a white solid. LCMS: (M+Hf: 477.1

Exarnpfe 8e 1-Ethyi-6.7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydrocinnoline-3- carbaidehyde

To a suspension of 1-ethyl-3-(hydroxymethyl)-6,7-bis((4-methoxybenzyl)oxy) cinnoiin-4(1 H)-one (1.6 g, 3.36 mmo!) In dich!oromethane (DCM) (50 mL) at room temperature was added Dess-Martin periodinane (1.709 g, 4.03 mmol). The white suspension turned into a dark green solution. The reaction was stirred at room temperature for 1 h, treated with sodium bicarbonate solution and extracted with dich!oromethane. The combined organic extracts were washed with brine, dried over sodium sulfate, filtered, and concentrated. The residue was purified by automatic silica gel column chromatography (Combiflash RF), eiuting with ethyl acetate/hexanes (0- 100%) to afford 1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoline-3- carba!dehyde (0.9 g, 1.897 mmol, 56.5 % yield) as a yellow solid. LCMS: (M+H)⁺: 475.2

Exampte 8f 1-Ethyi-6 J-bis((4-methoxybenzy!)oxy)-3-(Dyrroiidin-1 -ylmethyl)cinno!in- 4(1 H)-one

A solution of pyrrolidine (0.167 mL, 2.017 mmol) and 1-ethyl-6,7-bis((4- PU85023 methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoiine-3-carbaidehyde (0.87 g, 1.833 mmol) in 1 ,2-dichloroethane (DCE) (19 mL) and 2 drops of acetic acid was treated with sodium triacetoxyborohydride (0.583 g, 2.75 mmol). The reaction mixture was stirred for 1 .5 h and then concentrated. The residue was purified by automatic silica gei column chromatography (Combif!ash RF), eluting with [dicioromethane/methanoi/ammonium hydroxide(80:20:2)]/dichloromethane (0-60%) to afford 1-ethyl-6,7-bis((4- methoxybenzyl)oxy)-3-(pyrro!idin-1-yimethyl)cinnolin-4(1 H)-one (750 mg, 1.416 mmol, 77 % yield) as a light yellow gummy solid. LCMS: (M+H)⁺: 530.3

Exampte 8Q 1-(((6R,7R)-7-((E)-3-((((R)-1-(tert-butoxy)-4-((4-methoxybenzyl)oxy)-1 _!4- dioxobutan-2-y!)oxy)imino)-3-(2-((tenVbutoxyca^

(((4-methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2 ]oct-2-en-3-yl)methyi)-1 -

((1-ethyl-6J-bis((4-methoxybenzy!)oxy)-4-oxo-1 ,4-dihydrocinnolin-3-yl)me

1-ium

To the heat gun dried flask under nitrogen was added 1 -et yl-6,7-bis((4- methoxybenzyl)oxy)-3~(pyrro!idin-1-ylmethyi)clnnoiln-4(1 H)-one (750 mg, 1.416 mmol) in N.N-dimethylacetamide (DMA) (10 ml) followed by (S)-4-tert-butyl 1-(4-methoxybenzyl) 2-(((Z)-(1-(2-((tert-bLitoxycarbonyl)amino)thiazol-4-yl)-2-(((5R,6R7R)-3-(iodomethyi)-2- (((4-methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-7- yi)amino)-2-oxoethylidene)amino)oxy)succinate (Example 1 h) (1470 mg, 1.416 mmol) in Ν,Ν-dimethyiacetamide (DMA) (10 mL) . The mixture was heated at 40 °C for 3h, then cooled and stored in fridge overnight. The mixture was cooled to -40 °C and N,N~ dimethyiformamide (DMF) (10 mL) was added, foiiowed by phosphorus tribromide (0.267 mL, 2.83 mmol) dropwise over 0 min. Stirred at -40 "C for 30 min. Poured into the cooled solution of 5% sodium chioride. The resulting solid was filtered and dried, and chromatographed on ISCO silica gei column eluting with 0-20%

methanokdich!oromethane to give 1 -(((6R,7R)-7-((E)-3-((((R)-1-(tert-butoxy)-4-((4- methoxybenzyl)oxy)-1 ,4-dioxobutan-2-yi)oxy)imino)-3-(2-((tert- PU85023 butoxycarbonyi)amino)thiazoi-5-yl)-2-oxopropyi)-2-(((4-methoxybenzyi)oxy)carbonyi)-8- oxo-5-thia-1 -azabicyclo[4.2.03oct-2-en-3-yi)methyl)-1 -((1 -ethyi-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoiin-3-yl)methyi)pyrrolidin-1 -ium (1.35g , 0.948 mmol, 88.9 % yield). MS (M+H)⁺ 1426.4

Example 8fo 1 -(((6R,7R)-7-((E)-3-(2-aminothiazoi-5-yl)-3-(((R)-1 ,2-dicarboxyethoxy) imino)-2-oxopropyl)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yi)methyl)-1 -((1 - ethyl-GJ-dihydroxy^-oxo-l ^-dihydrocinno!in-S-y methy!Jpyrrolidin-l -ium, disodium salt

To a solution of 1 -(((6R,7R)-7-((E)-3-((((R)-1 -(tert-butoxy)-4-((4-methoxybenzyl) oxy)-1 ,4-dioxobutan-2-yl)oxy)imino)-3-(2-((tert-butoxycarbonyl)arnino)thiazoi-5-yi)-2- oxopropyi)-2-(((4-methoxybenzyl)oxy)carbonyi)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en- 3-yl)methyi)-1 -((1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoiin-3- yl)methyl)pyrrolidin-1 -ium (1 .35g , 0.948 mmol) in dichioromethane (DCM) (10 mL) was added anisole (3.1 1 mL, 28.4 mmol) followed by trifluoroacetic acid (3.85 mL, 47.4 mmoi). The resulting mixture was stirred at room temperature for 18h . Resulting mixture was triturated with isopropyi ether and filtered, and chromatographed on preparative HPLC column eluting with isocratlc solution of 10% acetonitrile and ammonium formate buffer to give desired Δ-3 Isomer. Desired compound was iyophiiized to constant weigh and acidified with 1 M hydrochloric acid. The solution was allowed to pass through the HP 20-ss column and then converted to disodium salt by adjusting the pH to 6.2 with 0.2M sodium hydroxide, and Iyophiiized to give 1 -(((6R,7R)-7-((E)-3-(2-aminothiazoi-5-yl)-3- (f(R)-1 ,2-dicarboxyethoxy)imino)-2-oxopropyl)-2-carboxy-8-oxo-5-th!a-1 - azabicyclo[4.2.0]oct-2-en-3-yl)methyl)-1 -((1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4- dihydrocinnolin-3-yl)methy!)pyrrolidin-1 -ium, disodium salt (52 mg, 0.082 mmol, 8.58 % yield) . LCMS (M+H)⁺ 788.4

¹ H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .31 - 1 .41 (m, 4 H) 2.02 (br. s„ 5 H) 2.51 - 2.61 (m, 3 H) 3.52 (br. s. , 5 H) 3.58 - 3.67 (m, 1 H) 3.87 (br. s. , 1 H) 4.20 (br. s., 1 H) 4.36 - 4.48 (m, 2 H) 4.51 (br. s. , 2 H) 4.76 (d, J=4.29 Hz, 1 H) 4.79 - 4.89 (m, 2 H) 5.26 (d, J=4.Q4 Hz, 1 H) 5.74 (d, J=4.55 Hz, 1 H) 6.73 (br. s„ 1 H) 6.90 (s, 1 H) 7.25 (s, 1 H) PU85023

Example 9 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((R)-1 ,2-dicarboxyet oxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]od-2-en-3-yl)methyi)-^ ethyl-BJ-dihydroxy^-oxo-l ^-dihydroquinoliri-S-y methylJpyrrolidin-l-ium, disodium salt

Exampte 9a 1-Ethyi-6_!7-bis((4-methoxvbenzvi)oxy)-3-(Dyrroiidin-1-ylmet yl)qui 4(1 H)-one

The titled compound was prepared according to Example 8a-f, utilizing 1~ethyl~ 6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3-carboxyi!c acid In place of 1 - ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoline-3-carboxyiic acid in Example 8a. LG S: (M+H)⁺: 529.3.

Example 9b 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl)oxy)-1 ,4- dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetam

(((4-methoxybenzyl)oxy)carbonyi)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yl)methyi)-1 - ((1-ethyi-67-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi)pyrroli 1-ium

To the heat gun dried flask under nitrogen was added 1 -ethyl-6,7-bis((4- rnethoxybenzyl)oxy)-3-(pyrrolidin-1-yimethyl)quinolin-4(1 H)-one (606 mg, 1.146 mmo!) in Ν,Ν-dimethy!acetamide (DMA) (10 mL), followed by (S)-4-tert-butyl 1-(4-methoxybenzy!) PU85023

2-(((Z)-(1-(2-((tert-butoxycarbonyl)amino)thiazol-4-yl)-2-(((5R,6R_!7R)-3-(iodomet yi)-2- (((4-methoxybenzyi)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-7- yi)amino)-2-oxoethylidene)amino)oxy)succinate (Example 1 h) (1 190 mg, 1.146 mmol) in Ν,Ν-dimethylacetamide (DMA) (10 mL) . Mixture was heated at 40 ^CiC for 3h, then cooled and stored in fridge overnight. The mixture was cooled to -40 "C and N,N~

dimethylformamide (DMF) (10 mL) was added, followed by phosphorus tribromide (0.216 mL, 2.293 mmol) dropwise over 10 mln. Stirred at -40 °C for 30 min and poured into the cooled solution of 5% sodium chloride. The resulting solid was filtered and dried, and chromatographed on ISCO silica gel column eiuting with 0-20%

methanokdichioromethane to give

1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzyl)oxy)-1 ,4-dioxobutan-2- yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-2-(((4- methoxybenzyl)oxy)carbonyl)-8-oxO-5 hia-1-azabicyc!o[4.2.0]oct-2-en-3-yl)methy!)-1 -((1- ethyl-6,7-bis((4-methoxyben2yi)oxy)-4-oxo-1 ,4-d!hydroquinollr>3-yl)methyi)pyrrolidln-1- ium (270 mg, 0.190 mmol, 16.54 % yield). LCMS (M+H 1424.4

Exampte 9c 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((R)-1 ,2-dicarboxyethoxy) lmino)acetamido)-2-carboxy-8-oxo-5 hia-1~azabicycio[4.2.0]oct-2-en~3-yl)methyl)~1-((1 ~ ethyi-6.7-dihydrQxy~4-oxo-1.4-dihydroquinoiin-3-yl)methyl)pyrrolidin-1-ium, disodium salt

To the solution of 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4- methoxybenzyl)oxy)-1.4-dioxobutan-2-yi)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)acetamido)-2-(((4-methoxybenzyi)oxy)carbonyi)-8-oxo- 5 hla-1-a2ab!cyclo[4.2.0joci-2-en-3-yl)methyl)-1-((1-ethyl-8,7-bis((4-methoxybenzyl)oxy)~ 4-oxo-1 ,4-di ydroquinolin-3-yi)methyl)pyrroiidin-1-ium (270 mg, 0.190 mmol) In dichioromethane (DCM) (2ml) was added anisole (0.207 ml, 1.897 mmol), followed by trifiuoroacetic acid (0.731 ml, 9.48 mmol) at room temperature. The mixture was stirred at room temperature for 18h. Resulting mixture was triturated with isopropyi ether and PU85023 filtered. Redisoived In aceionitrile (2mL) , water (1 mL) plus 2 drops of hydrochloric acid. Gilson HPLC eluting with 0-30% aceionitrile : water gave the fractions that were striped from acetonitrile and basified to pH 6.2 with 0.2M sodium hydroxide and lyophilized to give product 1-(((6R,7R)-7-((Z)-2-(2-arninothiazol-4-yi)-2-(((R)-1 ,2-dicarboxyethoxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]oGt-2-en-3-yl)methyi)-1-((1 - ethyi-BJ-dihydroxy^-oxo-l ^-dihydroquinoiin-S-y methyl pyrro!idin-l -ium disodium salt (28 mg, 0.034 mmo!, 17.73 % yield) as yellow solid. . LCMS: (M+H)⁺: 787.2

'H N R (400 MHz, DEUTERIUM OXIDE) δ ppm 1.38 (d, J=6.57 Hz, 3 H) 2.00 (br. s., 4 H) 2.48 - 2.61 (m, 3 H) 3.34 (br. s., 5 H) 3.86 (s, 1 H) 3.97 - 4.06 (m, 1 H) 4.16 - 4.25 (m, 1 H) 4.30 (br. s„ 2 H) 4.35 - 4.49 (m, 1 H) 4.78 - 4.89 (m, 2 H) 5.27 (d, J=4.80 Hz, 1 H) 5.74 (d, J=5.05 Hz, 1 H) 6.77 (br. s., 1 H) 6.84 - 6.93 (m, 1 H) 7.41 (s, 1 H) 8.06 (br. s., 1 H) 8.34 (s, 1 H)

Exaroplel O 1-(((6R,7R)~7-((Z)-2-(2~aminothiazol-4-yi)~2-(((2-carboxypropan~2-yl) oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yl)methyl)-1-

((1-ethyi-67-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)pyrroiidin-1 -ium

Example 10a 1-(((6R,7R)-2-((benzhydryloxy)carbony!)-7-((Z)-2-(((1-(tert-butoxy)-2- methyl-1-oxopropan-2-y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4- y!)acetamido)-8-oxo-5-thia-1 -azabicyclo[4.2 ]oct-2-en-3-yl)methyi)-1 -((1-ethyl-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoiin-3-yl)methyl)pyrrolidin-1-ium

To a solution of 1 -ethyi-6,7-bis((4-methoxybenzy!)oxy)-3-(pyrrol!din-1- y!methyi)cinnoiin-4(1 H)-one (Example 8f) (384mg, 0.725 mmoi) in N,N-dimethylaceiamide PU85023

(12 mL) under N2 at room temperature was added (6R ,7R)-benzhydryl 7-((Z)-2-(((1-(tert- butoxy)-2-methyl-1-oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4 yi)acetamido)-3-(!odornethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-erie-2-carboxyiate 5- oxlde (Example 6d) (812 mg, 0.870 mmo!) in N,N-dimethyiacetamide (6 mL). The mixture was stirred at room temperature over 12 h. LCMS indicated -31 % desired product in the crude mixture. N,N-dimethylforrnamide (32.00 mL) was then added. The mixture was cooled to -7Q°C and treated with phosphorous tribromide (0.150 mL, 1.595 mmo!) dropwise. The mixture was allowed to warm up to ~4Q°C. LCMS indicated complete reduction of the sulfoxide. The organic solution was poured into ice-cooled 5% sodium chloride (150 mL) and stirred for -15 min. The solid was separated by filtration, washed with water and dried under high vacuum to afford 1 -(((6R,7R)-2- ((benzhydry!oxy)carbonyl)-7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1-oxopropan-2- y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)am!no)thiazol-4-yi)acetamido)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-en-3-yi)methyl)-1-((1-et yl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo- 1 ,4-dihydrocinnolin~3-yl)nethy!)pyrroiidin-1 ~iun (1.45 g, 57% purity, 0.626 mmol, 86 % yield) as a dark brown solid. LCMS: (M+H)⁺: 1321.5.

Exarnpfe 10b 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl) oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-en-3-yl)methyl)-1- ((1-ethyi-6J-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)pyrroiidin-1 -ium

To a solution of 1 -(((6R,7R)-2-((benzhydryioxy)carbony!)-7-((Z)-2-(((1-(tert- butoxy)-2-methyl-1-oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4- y!)acetamido)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-((1-ethyl-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoiin-3-yl)methyi)pyrrolidiri-1 -ium (1.45 g, 0.626 mmol) (57% purity by LCMS) in dichloromethane (10 mL) under 2 at 0°C was added anisole (0.684 mL, 6.26 mmol), followed by trifluoroacetic acid (3 mL, 38.9 mmol) dropwise. The mixture was warmed up to room temperature and stirred at room temperature overnight. LCMS indicated completion of the reaction. Diisopropyl ether (20 mL) and water (4 mL) were then added. The mixture was rapidly stirred for -10 min and was then left standing for -10 min. The solvents were decanted away. The solid PU85023

(containing -70% of desired produci as shown by LCMS) was dissolved in a mixed solvents of water (10 mL), acetonitriie (10 mL) and 2N aqueous hydrochloric acid (2.5 mL), and was then absorbed onto HP20ss resin, and purified by reverse phase chromatography using HP20ss pre-column followed by 40 g C18 column, eluting with 0- 20% acetonitrile/water. Only two fractions containing -73% of desired product were obtained. The two fractions were combined, concentrated under reduced pressure to remove acetonitriie, frozen and placed on lyophilizer for 24 hours to provide 101 mg of product (~72% pure by LCMS). This product was dissolved in acetonitriie (5 mL) and 1 N hydrochloric acid (5 mL) and purified by Gilson automated HPLC (5-60% organic, 8 min gradient) to afford after lyophilization 1 -(((6R_>7R)-7-((Z)-2-(2-aminothiazo!-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct- 2-en-3-yi)methyl)-1 -((1 -ethyi-6_>7-dihydroxy-4-oxo-1 _l4-dihydrocinnolin-3- yi)methyl)pyrroiidin-1-ium (40 mg, 0.050 mmol, 7.93 % yield) ( 00% pure by LCMS, 94% by HPLC). LCMS: (M+H)*: 758.4. 1 _H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.30 - 1.48 (m, 9 H) 2.08 (br. s., 4 H) 3.35 -3.70 (m, 6 H) 3.80 - 3.98 (m, 1 H) 4.1 1 - 4.28 (m, 1

H) 4.53 (br. s., 4 H) 5.30 (br. s., 1 H) 5.78 (d, J=4.04 Hz, 1 H) 6.89 (s, 1 H) 6.99 - 7.18 (m, 1 H) 7.28 - 7.45 (m, 1 H).

E ample 11 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl) oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-y!)methyl)-1- (2-(1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoiine-3-carboxamido)ethyl)pyrroi!din-1 -ium

Example 11 a 1-Eth !idin-1-yl)ethyl)- 1 ,4-dihydrocinnoline

To a suspension of 1-ethyl-6.7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4- ocinnoline-3-carboxylic acid (Example 8c) (2.5 g, 5. 0 mmol) in dichioromethane (50 mL) at room temperature was added thionyi chloride (0.446 mL, 6.12 mmol), and the mixture was heated to reflux for 1 h. After cooling to room temperature the mixture was added dropwise to a solution of 2-(pyrrolidin-1 -yi)ethanamine (0.71 1 mL, 5.81 mmol) and triethylamine (1 ,086 mL, 7.65 mmol) in dichioromethane (50.0 mL) in an ice bath . The resulting mixture was then allowed to warm up to room temperature, and stirred for another 0.5 h. LC S indicated completion of the reaction. The mixture was

concentrated in vacuo, and the residue was purified twice by automatic silica gel column chromatography (Combifiash RF), eluting with a mixture of

[dichloromethane/methanoi/ammonium hydroxide(80:20:2)]/dichloromethane (0-50%) to afford 1 -ethyl-6.7-bis((4-methoxybenzyi)oxy)-4-oxo-N-(2-(pyrroiidin-1 -yl)ethyl)-1 ,4- dihydrocinnoline-3-carboxamide (2.2 g, 3.75 mmol, 73.8 % yield) as a light yellow solid . LCMS: (M+Hf: 587.3.

[Example 11 1 -(((6R,7R)-7-((Z)-2-(2-aminothiazo!-4-yl)-2-(((2-carboxypropan-2-y!) oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-en-3-yl)methyl)-1 - (2-(1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoiine-3-carboxamido)ethyl)pyrroiidin-1 -ium

The compound was prepared according to the two-step sequence of Examples 10a-10b, utilizing the compound from Example 1 1 a and the compound from Example 8d to afford 1 -(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yl)methyl)- 1 -(2-(1 -ethyl-6 J-dihydroxy^-oxo-l ^-dihydrocinnoiine-S-carboxamidoiethylJpyrro!idin-l - ium (the combined yield for the two steps: 1 .93% ). LCMS: ( +H)⁺: 814.7. ¹ H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .31 - 1 .45 (m, 9 H) 2.13 (br. s. , 4 H) 3.58 (br. s., 6 H) 3.75 - 4.18 (m, 4 H) 4.49 (d, J=7.58 Hz, 4 H) 5.28 (d, 5.05 Hz, 1 H) 5.76 (s, 1 H) 6,93 (s, 1 H) 7.10 (s, 1 H) 7.37 (s, 1 H).

Example 12 1 -(((6R,7R)~7-((Z)-2-(2~aminothiazol-4-yi)~2-(((2-carboxypropan~2-yl) oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-y!)methyi)-1 -(2- (1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido)ethyi)pyn lidin-1 -ium PU85023

The compound was prepared according to Examples 8a-c and 1 1 a, utilizing 5- ethyi-8-oxo-5,8-dihyd! -[1 ,3]dioxoio[4,5-g]quino!ine~7-carboxylic acid in place of 1~ethyi~4- oxo-1 ,4-dihydro-[1 ,3]dioxQio[4,5-g]cinno!ine-3-carboxy!ic acid in Examples 8a. LCMS: ( +Hf: 586.3.

Examj3feJ2b 1-(((6RJR)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azab^

ethyl-67-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxam

The compound was prepared according to the two-step sequence In Examples I Qa- Ob . utilizing the compound from Example 12a and the compound from Example 6d to provide 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yl)methyl)- 1-(2-(1 -ethyl-6 ,7-dihydroxy-4-oxo-1 ,4-di ydroquinol!ne-3-carboxamido)ethyi)pyrrolidin-1- lum (the combined yield for the two steps: 0.95% ). LCMS: ( +H)⁺: 814.2. ¹ H NMR (400 MHz, DEUTERIUM OXIDE} δ ppm 1.32 (br. s., 9 H} 2.03 - 2.22 (m, 4 H) 3.31 - 4.46 (m, 14 H) 5.19 - 5.33 (m, 1 H) 5.70 - 5.82 (m, 1 H) 6.80 - 6.92 (in, 2 H) 7.41 - 7.54 (in, 1 H) 8.44 - 8.60 (in, 1 H) PU85023

Example 13 1 -(((6R,7R)-7-((E)-3-(2-aminothiazoi-5-yl)-3-(((R)-1 ,2-dicarboxyethoxy) imino)-2-oxopropyl)-2-carboxy-8-oxo-5-th!a-1-azabicyclo[4.2.0]oct-2-en-3-yl)methy ethyl-6 J-dihydroxy^-oxo-l ^-dihydrocinnoiin-S-y methy piperidin-l-ium, disodium salt

Exam le 13a 1 -ethyl-6, 7-bis((4-methoxybenzyl)oxy)-3-(piperidin-1 -yimethyi)ci

4(1 H)-one

To a suspension of 1-ethyl-3-(hydroxymethyl)-6,7-bis((4- methoxybenzyl)oxy)cinnoiin-4(1 H)-one (Example 8d) (2.13 g, 4,47 mmol) in

Dichloromethane (DCM) (40 mL) In an ice bath under N2 was added TEA (1.495 mL,

10.73 mmol) followed by methanesulfonyl chloride (0.627 mL, 8.05 mmol) (white suspension turned into a light yellow clear solution). The mixture was stirred in an ice bath for 2 h, and then the above reaction mixture was added dropwlse into a stirred solution of piperidine (1.195 mL, 12.07 mmol) in dichloromethane (DCM) (40.0 mL) at room temperature overnight. The reaction mixture was concentrated and purified through column chromatography e!uting with dichloromethane and methano!/dichioromethane /ammonium hydroxide (from 0% to 100%) over 15 mins twice to afford 1 -ethy!-6,7-bis((4- methoxybenzyl)oxy)-3-(piperidin-1 -ylmethyi)ciririoiin-4(1 H)-one (1.54 g, 2.55 mmol, 57.0

% yield). LCMS (M+H)⁺ : 544.0

Example 13b 1-(((6R,7R)-7-((E)-3-((((R)-1-(tert-butoxy)-4-((4-methoxybenzyl)oxy)-1 _!4- dioxobutan-2-yl)oxy)imino)-3-(2-((tert-butoxycarbonyl)amino)thiazol-5-yi)-2-oxop methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yi)methyl)-1 -((1- ethyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydrocinnolin-3-yl)methy!)piperidin-1-ium PU85023

To the heat gun dried flask under nitrogen was added 1 -ethyl-6,7-bis((4-methoxy benzyl) oxy)-3-(piperidin-1-ylmethy!)cinnoiin-4(1 H)-one (610 mg, 1 .122 mmol) in N,N- dimethylacetamide (DMA) (10 mL) followed by (S)-4-tert-butyi 1 -(4-methoxybenzyi) 2- (((Z)-(1 -(2-((tert-butoxycarbonyl)amino)thiazoi-4-yl)-2-(((5R,6RJR)-3-(!odomethyl)-2-(((4- methoxybenzyl)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-en-7- yl)amino)-2-oxoethylidene)amino)oxy)succinate ( 165 mg, 1.122 mmol) in N,N- dimethylacetamide (DMA) (10 mL) . Mixture was heated at 40 ^!,C for 3h, cooled and stored in fridge overnight. THe mixture was cooled to -40 °C and N,N-dimethylformamide (DMF) (10 mL) was added followed by phosphorus tribromide (0.212 mL, 2.24 (mmol) dropwise over 10 min. Stirred at -40 "C for 30 min, and poured into the cooied solution of 5% sodium chloride. The resulting solid was filtered and dried, and chromatographed on ISCO silica gel column eluting with 0-20% methanokdich!oromethane to give 1 - (((6R,7R)-7-((E)-3-((((R)-1-(tert-butoxy)-4-((4-methoxybenzyl)oxy)-1 ,4-dioxobutan-2- yi)oxy)imino)-3-(2-((tert-butoxy carbonyi)amino)thiazol-5-yi)-2~oxopropyl)-2~(((4~

methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yi)methyi)-1 -((1- ethyi-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihy

(640 mg, 0.445 mmol, 39.6 % yield) . LCMS ( +H)⁺: 1439.4 Example 13 c 1 -(((6R,7R)-7-((E)-3-((((R)-1 -(tert-butoxy)-4-((4-methoxybenzy!)oxy)-1 ,4- dioxobutan-2-y!)oxy)imlno)-3-(2-((tert-butoxy

methoxybenzyl) oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yl)methyl)-1-((1-ethyi- 6 J-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)piperidin-1-ium PU85023

To the solution of 1 -(((6R,7R)-7-((E)-3-((((R)-1 -(tert-butoxy)-4-((4-methoxybenzyl) oxy)-1 ,4-dioxobutan-2-yl)oxy)imino)-3-(2-((tert-butoxycarbonyl)amino)thiazoi-5-yl)-2- oxopropyi)-2-(((4-methoxybenzy!)oxy)carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en- 3-yl)methyi)-1 -((1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnolin--3- y!)methyl)piperidin-1-ium (840 mg, 0,445 mmoi) In dichloromethane (DCM) (10 mL) was added anisole (1 .458 mL, 13.35 mmol) and trifiuoroacetic acid (1.714 mL, 22.24 mmol) at room temperature. The reaction mixture was stirred at room temperature for 8h.

Resulting mixture was triturated with isopropyl ether and filtered. The solid was redissolved in acetonitrile (2mL) , water (1 mL) plus 2 drops of hydrochloric acid and purified through ISCO C-18 column and HP-2Qss pre-column e!uting with 0-30% acetonitrile : water. The fractions were concentrated to remove acetonitrile and basifled to pH 8.2 with 0.2 sodium hydroxide and lyophllized to give product 1-(((6R,7R)-7-((E)-3- (2-aminothiazoi-5-yl)-3-(((R)-1 ,2-dicarboxyethoxy)imino)-2-oxopropyi)-2-carboxy-8-oxo-5- thia-1 -azabicyclo[4.2.0]oct-2-en-3-y!)rnethyl)-1-((1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4- dihydrocinnolin-3-yi)methy!)piperidin-1-ium, disodium salt (35 mg, 0.041 mmol, 9.28 % yield) LCMS (M+H)⁺:802.3

Ή N R (400 MHz, DEUTERIUM OXIDE) δ ppm 1.23 - 1.49 (m, 4 H) 1.53 (br. s., 2 H) 1.71 - 1.84 (m, 2 H) 1.82 (d, J=1 1.62 Hz, 2 H) 2.02 (br. s., 1 H) 2.40 - 2.71 (m, 4 H) 2.53 - 2.63 (m, 2 H) 3.07 - 3.20 (m, 1 H) 3.28 (br. s., 2 H) 3.43 (d, J=16.93 Hz, 2 H) 3.87 (d,

J=16.93 Hz, 1 H) 4.15 (d, J-14. 5 Hz, 1 H) 4.44 (d, J-7.07 Hz, 2 H) 4.57 (br. s., 1 H) 4.80 (br. s., 2 H) 4.85 (dd, J-9.09, 4.29 Hz, 2 H) 5.28 (d, J=5.05 Hz, 1 H) 5.75 (d, _/=4.80 Hz, 1 H) 8.68 (s, 1 H) 6.83 - 8.93 (m, 1 H) 8.86 - 8.97 (m, 1 H) 7.27 (s, 1 H)

Example 14 1 -(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)meth

1-(2-(3-hydroxy-1 -methyi-4-oxo-1 ,4-dihydroquinoline-8-carboxamido)ethyi)pyrrolidin-1- ium PU85023

Exampte 14a Ethyl 2-(((2-bromophenyi)amino)methylene)-3-oxobutanoate

To a solution of 2-bromoaniline (2 g, 1 1.83 mmo!) In Isopropanol (15 mL) stirred under nitrogen at room temperature was added neat ethyl 2-(ethoxymethylene)-3- oxobutanoate (2.185 g, 1 1.83 mmoi) in one charge. The reaction mixture was stirred at room temperature for 20 min, at which time LCMS showed only a trace amount of product. The reaction mixture was heated to 75 °C and stirred for 20 min. LCMS showed complete conversion. The mixture was cooled to room temperature and concentrated under vacuum, during which time a large amount of material precipitated out. Filtration followed by a hexanes wash afforded ethyl 2-(((2-bromophenyl)amino)methyiene)-3- oxobutanoate (3.8 g, 10.98 mmo!, 94% yield) that was apx. 90% pure to be used in the next step. LCMS: (M+H)⁺: 312.0.

To Dowtherm (20 mi) at apx 200°C was added solid ethyl 2-(((2-bromophenyi) amino) methylene)-3-oxobutanoate (3 g, 9.61 mmol). The reaction was heated to 250 °C and stirred for 1 h. LCMS showed consumption of starting material and formation of desired product along with an unknown biproduct of lower mass. The reaction was allowed to cool to room temperature and was them poured into 150 mL of hexanes. The mixture was stirred for 15 min, then the precipitate was collected by filtration and dried to afford 3-acetyi-8-bromoquinolin-4(1 H)-one (1 .77 g, 6.65 mmoi, 69.2 % yield) as a light brown solid. LCMS: (M÷H)⁺: 285.9. !θ: 3

Exam pie 14c 3-Acetyl-8-bromo-1 -met yiquinoiin-4(1 H)-one

AA mmiixxttuurree ooff 33--aacceettyyll--88--bbrroommooqquuiinnoolliinn--44((11 HH))--oonnee ((55..3355 gg,, 2200..11 11 mmmmooii)) aanndd KK₂₂CC00₃₃ ((99..1177 gg,, 8888..33 mmmmoo!l)) iinn ddrryy NN,,NN--ddiimmeetthhyyllffoorrmmaammiiddee ((110000 mmLL)) wwaass hheeaatteedd aatt 9900 ^tt!!CC ffoorr 3300 mmiinn.. AAfftteerr ccoooolliinngg ttoo rroooomm tteemmppeerraattuurree,, iiooddoommeetthhaannee ((33..7777 mmLL,, 6600..33 mmmmooli)) wwaass aaddddeedd aanndd tthhee mmiixxttuurree wwaass ssttiirrrreedd aaggaaiinn aatt 9900 ""CC f foorr 22 hh.. TThheenn tthhee rreeaaccttiioonn mmiixxttuurree wwaass ccoonncceennttrraatteedd aanndd ddiilluutteedd wwiitthh wwaatteerr ((5500 mmLL)),, eexxttrraacctteedd wwiitthh ddiicchhlloorroommeetthhaannee ((44 xx 5500 mmLL)),, ddrriieedd oovveerr ssooddiiuumm ssuullffaattee aanndd aabbssoorrbbeedd oonnttoo ssiilliiccaa ggeell.. PPuurriiffiiccaattiioonn bbyy aauuttoommaattiicc ssiilliiccaa ggeeii ccoolluummnn cchhrroommaattooggrraapphhyy ((CCoommbbiiff!!aasshh RRFF)) eelluuttiinngg wwiitthh eetthhyyll aacceettaattee//hheexxaanneess ((1100--110000%%)) aaffffoorrddeedd 33--aacceettyyll--88--bbrroommoo--11--mmeetthhyyiiqquuiinnooiiiinn--44((11 HH))--oonnee ((33..44 gg,, 1122..1144 mmmmooii,, 6600..44 %% yyiieelldd)) aass aa bbrroowwnn ssoolliidd.. LLCCMMSS:: ((MM++HH))⁺⁺:: 227799..99..

88--BBrroommoo--11--mmeetthhyyll--44 ,,44--ddiihhyyddrrooqquuiinnooiiiinn--33--yyll aacceettaattee

TToo aa ssoolluuttiioonn ooff 33--aacceettyyll--88--bbrroommoo--11--mmeetthhyyllqquuiinnoolliinn--44((11 HH))--oonnee ((33..44 gg,, 1122..1144 mmmmooil)) iinn ddiicchhlloorroommeetthhaannee ((6600 mmLL)) aatt rroooomm tteemmppeerraattuurree wwaass aaddddeedd ppoorrttiioonnwwiissee wwiitthh ssttiirrrriinngg mm-- GGPPBBAA ((44..0088 gg,, 88..2211 mmmmoo!l)).. TThhee rreessuullttiinngg mmiixxttuurree wwaass ssttiirrrreedd aatt rroooomm tteemmppeerraattuurree ffoorr 11 hh.. TThhee eexxcceessss mm--CCPPBBAA wwaass qquueenncchheedd vviiaa aaddddiittiioonn ooff 1155%% aaqquueeoouuss NNaa₂₂SS00₃₃ ((3300 mmLL)),, aanndd tthhee rreessuullttiinngg mmiixxttuurree wwaass ssttiirrrreedd aatt rroooomm tteemmppeerraattuurree ffoorr 00..55 hh.. TThhee llaayyeerrss wweerree sseeppaarraatteedd,, aanndd tthhee oorrggaanniicc llaayyeerr wwaass wwaasshheedd sseeqquueennttiiaallllyy wwiitthh wwaatteerr ((110000 mmLL)),, 55%% aaqquueeoouuss NNaaHHCCOO₃₃ (( 0000 mmLL)),, aanndd bbrriinnee ((110000 mmLL)).. TThhee oorrggaanniicc llaayyeerr wwaass ddrriieedd ((NNaa₂₂88CC>>44)),, ffiilltteerreedd,, aanndd ccoonncceennttrraatteedd iinn vvaaccuuoo ttoo aaffffoorrdd ccrruuddee 88--bbrroommoo--11--mmeetthhyyii--44--ooxxoo--11 ,,44-- ddiihhyyddrrooqquuiinnooiiiinn--33--yyll aacceettaattee ((33..6688 gg,, 1100..5566 mmmmooll,, 8877 %% yyiieelldd)) aass bbrroowwnn ssoolliidd,, wwhhiicchh wwaass uusseedd iinn tthhee nneexxtt hhyyddrroollyyssiiss sstteepp wwiitthhoouutt ffuurrtthheerr ppuurriiffiiccaattiioonn.. LLCCMMSS:: ((MM++HH))^**:: 229955..99..

PU85023

To a suspension of 8-bromo-1-methyl-4-oxo-1 ,4-dihydroquinolin-3-yl acetate (3.88 g, 10.56 mmoi) in neihanoi (100 mL) was added aqueous 2 M KOH (15 mL), the mixture turned into a brown solution and was stirred at room temperature for 1 h. The reaction mixture was concentrated, and was then partitioned between dichiorometnane and brine. The organic phase was separated, the aqueous phase was extracted with

dichiorometnane for several times. The combined organic extracts were dried ( a₂S04), filtered and the solvent was removed in vacuo. The residue was then absorbed onto silica gel and purified by automatic silica gel column chromatography (Combifiash RF), e!uting with MeOH/ dichioromethane ( 0-5%) to afford 8-bromo-3-hydroxy-1 -methylquinolin- 4(1 H)-one (2.47 g, 9.24 mmol, 87 % yield) as a brown solid. LCMS: (M+H)⁺: 254.1

Exampje 14f 8-Bromo-3-((4-methoxybenzyl)oxy)-1 -methylquinolin-4(1 H)-one

A suspension of 8-bromo-3-hydroxy-1-methylquinolin-4(1 H)-one (2.4 g, 8.97 mmol) and potassium carbonate (1.488 g, 10.77 mmol) in Ν,Ν-dimethy!formamide (50 mL) was heated at 90 °C for 0.5 h. Then 1-(chioromethyi)-4-methoxybenzene (1.442 mL, 9.87 mmoi) and potassium iodide (0.149 g, 0.897 mmol) were added, and the mixture was heated at the same temperature for another 2 h. After cooling, N,N- dimethyiformamide was removed in vacuo and the residue was partitioned between dichioromethane and brine. The organic phase was separated, the aqueous phase was extracted with dichioromethane for several times. The combined organic extracts were dried over sodium sulfate, filtered and concentrated, The residue was purified by automatic silica gel column chromatography (Combifiash RF), eluting with ethyl aceiate/hexanes (0-100%) to afford 8-bromo-3-((4-methoxybenzyl)oxy)-1-methylquinolin- 4(1 H)-one (2.8 g, 6.95 mmoi, 77 % yield) as a light brown solid. LCMS: (M+H)⁺: 374.0.

Exampte 14q 3-((4- ethoxybenzyi)oxy)-1-methyi-4-oxo-1 ,4-dihydroquinoline-8- carbonitriie PU85023

To a suspension of 8-bromo-3-((4-methoxybenzyl)oxy)-1 -methy!quino!in-4(1 H)- one (2.6 g, 6,95 mmol), DPPF (0.462 g, 0.834 mmol), palladium ieirakis (0.482 g, 0.417 mmo!) and Pd₂(dba)₃ (0.382 g, 0.417 mmol) in Ν,Ν-dimethylformamide (30 mL) at room temperature was added dicyanozinc (0.979 g, 8.34 mmol). The mixture was then heated at 1 10 °C under N₂ for 1 h. After cooled to room temperature, the mixture was concentrated in vacuo and then partitioned between EtOAc (80 mL) and cone. NH₄GI (40 mL). The organic phase was separated, washed with brine (40 mL), dried over sodium sulfate, and concentrated. The residue was purified twice by automatic silica gel column chromatography (Comblflash RF), e!uting with ethyl acetate/ hexanes (0-100%) to afford 3-((4-methoxybenzyl)oxy)-1 -methyi-4-oxo-1 ,4-dihydroquinoline-8-carbonitriie (1.87 g, 5.84 mmol, 84 % yield) as a light brown solid. LCMS: ( +H)^' . 321 .1 .

Example 14 3-((4-Methoxybenzyi)oxy)-1-methyi-4-oxo-1 ,4-dihydroquinoline-8- carboxamide

A solution of 3-((4-methoxybenzyl)oxy)-1-methyl-4-oxo-1.4-dihydroquinoline-8- carbonitriie (1.75 g, 5.48 mmol) and 25% aqueous sodium hydroxide (20 mL) in methanol (100 mL) was stirred at reflux for 3 h. After complete consumption of the nitriie, the mixture was concentrated to remove -75% of the methanol. The mixture was acidified to pH 3 with 1 M HCI (aq), and partitioned between EtOAc and brine. The organic phase was separated, dried over sodium sulfate and concentrated in vacuo. The crude product was absorbed onto silica gel and purified by automatic silica gel column chromatography (Combiflash RF), eluting with methano!/dichlorometharie (0-30%) to afford 3-((4- methoxybenzyl)oxy)-1-methyl-4-oxo-1 ,4-diriydroquino!ine-8-carboxamide (1 .21 g, 3.58 mmol, 85.5 % yield) as light brown solid. LCMS (M+H)⁺: 339.3.

Exampte 14j: 3-((4-methoxybenzyi)oxy)-1-methyi-4-oxo-N-(2-(pyrrolidin-1 -y!)ethyi)-1 ,4- dihydroquinoiine-8-carboxamide PU85023

TToo aa ssoolluuttiioonn ooff 33--((((44--mmeetthooxxyybbeennzzyyii))ooxxyy))--11 --mmeetthyyii--44--ooxxoo--11 ,,44--ddiihhyyddrrooqquuiinnoolliinnee--88-- ccaarrbbooxxaammliddee ((11..0099 gg,, 33..2222 mmmmoo!i)) iinn NN,,NN--ddiimmeetthhyy!iffoorrmmaammiiddee ((3300 mmLL)) wwaass aaddddeedd ssooddiiuumm hhyyddrriiddee ((00..119933 gg,, 44..8833 mmmmoo!i)) aanndd tthhee mmiixxttuurree wwaass ssttiirrrreedd aatt rroooomm tteemmppeerraattuurree ffoorr 11 hh..

55 TThheenn tthhee mmiixxttuurree ooff 11 --((22--cchh!!oorrooeetthhyyll))ppyyrrrroolliiddiinnee,, HHyyddrroocchhlloorriiddee ((00..554488 gg,, 33..2222 mmmmooii)) aanndd ttrriieetthhyyllaammiinnee ((00..449944 mmLL,, 33..5544 mmmmooii)) iinn NN,,NN--ddiimmeetthhyyiiffoorrmmaammiiddee ((3300..00 mmLL)) wwaass aaddddeedd IInn oonnee ppoorrttiioonn.. TThhee rreessuullttiinngg mmiixxttuurree wwaass ssttiirrrreedd aatt 5500 ""CC ffoorr 33..55 hh.. EExxttrraa NNaaHH ((00..1155 eeqq)) wwaass aaddddeedd,, tthhee mmiixxttuurree wwaass ssttiirrrreedd aatt 5500 °°CC ffoorr aannootthheerr 00..55 hh.. TThhee ssoollvveenntt wwaass rreemmoovveedd iinn vvaaccuuoo,, aanndd tthhee rreessiidduuee wwaass ppaartrtiittiioonneedd bbeettwweeeenn wwaatteerr ((5500 mmLL)) aanndd ddiicchhlloorroommeetthhaannee ((5500 1100 mmLL)),, aanndd eexxttrraacctteedd wwiitthh DDCCMM ffoorr tthhrreeee ttiimmeess.. TThhee ccoommbbiinneedd oorrggaanniicc pphhaasseess wweerree ddrriieedd

oovveerr ssooddiiuumm ssuullffaattee,, ffiilltteerreedd,, aanndd ccoonncceennttrraatteedd.. TThhee l liigghhtt yyeellllooww rreessiidduuee wwaass ppuurriiffiieedd bbyy aauuttoommaattiicc ssiilliiccaa ggeell ccoolluummnn cchhrroommaattooggrraapphhyy ((CCoommbbiiffllaasshh RRFF)),, eeiiuuttiinngg wwiitthh

[[GCHH₂sCC!i₂s//MMeeOOHH//NNHHj,ODHH ((8800::2200::22))]]// CCHH₂₂CCII₂₂ ((00--5500%%)) ttoo aaffffoorrdd 33--((((44--mmeetthhooxxyybbeennzzyyll))ooxxyy))--11-- mmeetthhyyll--44--ooxxoo--NN--((22--((ppyyrrrroolliiddiinn--11--yyii))eetthhyyll))--11 ,,44--ddiihhyyddrrooqquuiinnooiliinnee--88--ccaarrbbooxxaammiiddee ((00..33 gg,, 00..888899 1155 mmmmoo!!,, 2211..3388 %% yyiieelldd)) aass aa lliigghhtt bbrroowwnn ooiill.. LLCCMSS:: (( ++HH))⁺⁺:: 443366..33..

11 --((((((66RR,,77RR))--22--((((bbeennzzhhyyddrryyllooxxyy))ccaarrbboonnyyll))--77--((((ZZ))--22--((((((11--((tteerrtt--bbuuttooxxyy))--22--mmeetthhyyil--11-- oxOpropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thia2ol-4-yl)acetamido)-8~oxo-5- th!a-1 ~a2abicyclo[4.2.0]oct-2-en~3-y!)methyl)-1 -(2-(3-((4-methoxybenzyl)oxy)~1-methyl-4-oxo- 20 1 ,4-dihydroquinoiine-8-carboxamido)ethyl)pyrroiidin-1-ium

To a solution of 3-((4-methoxybenzyl)oxy)-1-methyi-4-oxo-N-(2-(pyrrolidin-1- y!)ethyi)-1 ,4-dlhydroquinoline-8-carboxamide (0.202 g, 0.464 mmoi) in N,N- dimethylacetamide (4 mL) under 2 was added (6R,7R)-benzhydryl 7-((Z)-2~(((1-(tert~ butoxy)-2-methyl-1~oxopropan-2-yl)oxy)lmlno)-2-(2-((tert-butoxycarbonyl)amino)thia2ol-4- yl)acetamido)-3-(iodomethyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2-carboxylate 5- PU85023 oxide (0.520 g, 0.557 mmo!) in N,N-dimethylacetamide (4 mL). The mixture was stirred at room temperature over night. LCMS indicated -18% of desired product in the reaction mixture. N,N-Dimethy!formamide (18.00 mL) was then added. The mixture was cooled to -70°C and treated with tribromophosphine (0.096 mL, 1 .020 mmol) dropwlse. The mixture was allowed to warm up to -40°C. LCMS Indicated complete reduction of sulfoxide. The organic solution was poured into ice-cooled 5% sodium chloride (80 mL) and stirred for -15 mln. The solid was separated by filtration, washed with water and dried under high vacuum to afford a dark brown solid (0.45 g, contained 42% of desired product by LCMS). The crude product was further purified by Combiflash automated silica gei chromatography (12 g Gold column, 0-15% MeOH in DCM) to provide 1 -(((6R,7R)-2- ((benzhydry!oxy)carbonyl)-7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1-oxopropan-2- y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-8-oxo-5-th azabicyclo[4.2 ]oct-2-en-3-yi)methyl)-1-(2-(3-((4-methoxybenzyl)oxy)-1-methyi-4-oxo- 1 ,4-dihydroquinoiine-8-carboxamido)ethyl)pyrroi!din-1 -ium (66 mg, 80% purity, 0.043 mmol, 9.28 % yield) as a brown solid. LCMS: (M+H)⁺: 1226.0.

Exarnpfe 14S: 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycloE4.2.0]oct-2-en-3-yl)meth

(3~hydroxy-1-methyi-4-oxo-1 ,4-dlhydroquinoline-8-carboxamido)ethyi)pyrrolidin-1-lum

To a solution of 1 -(((6R,7R)~2-((benzhydryioxy)carbony!)-7-((Z)-2-(((1-(tert- butoxy)-2-methyl-1-oxopropan-2-yl)oxy)imlno)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4- yl)acetamido)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-(2-(3-((4- methoxybenzyl)oxy)-1-methyl-4-oxo-1 ,4-dihydroquinoiine-8-carboxamido)ethyl)pyrroiidin- 1-ium (66 mg, 0.054 mmol) in dichioromethane (2 mL) under at 0°C was added anisole (0.059 mL, 0.538 mmol), followed by trif!uoroacetic acid (0.5 mL, 8.49 mmol). The mixture was warmed up to room temperature and stirred over night. LCMS indicated completion of the deprotection. Dilsopropyl ether (5 mL) and water (1 mL) were then added. The mixture was stirred for 10 mln and the solvents were decanted away from the solid. The solid was dissolved in acetonitrlle (2 mL) and 1 N HCI (2 mL), and purified by PU85023

Gilson automated HPLC (acetonitrile/Water: 5-65%). The pure fractions were combined and concentrated under vacuum to -15 mL (bath temperature 20°C) and freeze-dried to afford 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- yi)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yl)methyl)- 1-(2-(3-hydroxy-1 -methyi-4-oxo-1 ,4-dihydroquinoline-8-carboxamido)ethyi)pyrrolidir!-1- ium (5 mg, 84% purity, 5.36 μιηοί, 9,96 % yield) as a yellowish solid. LCMS: (M+H)⁺:

783.2. ¹ H NMR (400 MHz, METHANOL-d4) δ ppm 1.56 - 1.70 (m, 6 H) 2.30 (br. s., 4 H) 3.62 - 4.31 (m, 14 H) 4.42- 4.58 (m, 1 H) 5.38 - 5.50 (m, 1 H) 5.89 - 6.05 (m, 1 H) 7.12 - 7.28 (m, 1 H) 7.62 - 7.77 (m, 1 H) 7.89 - 8.05 (m, 1 H) 8.32 - 8.49 (m, 1 H) 8.54 - 8.69 (m, 1 H)

Example 15 (6R_!7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-((1 -((1 -ethyl-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin- 3-yl)methyi)pyrrolidin-1-ium-1 -yi)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, Sodium salt

A mixture of 3-fiuorobenzene-1 ,2-dioi (145 g, 1 132 mmo!), potassium carbonate (313 g, 2264 mmol) and iodomethane (177 mL, 2830 mmol) in Ν,Ν-Dimethyiformamide (D F) (300 mL) was stirred at RT for 48h. LCMS showed the complete consupiion of starting material. The mixture was diluted with ethyl acetate (EA), filtered, washed with water, dried and evaporated in vacuo. The crude material was purified by normal phase automatic silica gel column chromatography (Combiflash RF, 220g column), eluting with PU85023

EA/Hexane (10%-80%) to afford 1 -fiiioro-2,3-dimethoxybenzene (149 g, 954 mmoi, 84 yield) as a colorless oil. LCMS (M+H)⁺: 157.0.

Example 15b 2-Fluoro-3,4-dimethoxy-1 -nitrobenzene

With sirring, to ice cold nitric acid (853 ml, 1.9 E+04 mmoi) was added dopwise 1 -fiuoro- 2,3-dimethoxybenzene (149 g, 954 mmoi). The mixture was stirred at 0 °C for another 15mins and then warmed up to RT for 15mins. The orange solution was poured onto ice and the resultant solid was filtered, washed with water and dried. LCMS showed the pale yellow solid was the mixture of 6-nitro and 5-nitro product (ratio = 1/1.6). The crude material was purified by reverse phase automatic C18 column chromatography

(Combifiash RF, 120g column) for several times, eluting with Acetonitrile/ Water (0-70%) to afford 2-fluoro-3,4-dimethoxy-1-nitrobenzene (53 g, 263 mmoi, 27.6 % yield) as a white solid. LCMS (M+H)⁺: 202.1.

Example 15c 2-Fiuoro-3,4-dimethoxyaniline

2-Fluoro-3,4-dimethoxy-1-nitrobenzene (52 g, 259 mmoi) was hydrogenated on a Par shaker (30 psi) in Ethanoi (300 mL) with platinum(IV) oxide (5 g, 22.02 mmoi) at r.t. for 30 mins. LCMS indicated completion of the reaction. The catalyst was filtered off and the filtrate was concentrated to afford 2-fiuoro-3,4~dimethoxyaniilne (42 g, 245 mmoi, 95 % yield) as a brown oil. LCMS (M+H)⁺: 72.0.

Example 15d 2-(((2-Fluoro-3,4-dimethoxyphenyl)amino)methyiene)malonate

To a solution of 2-fiuoro-3,4-dimethoxyaniiine (39 g, 228 mmol) in Ethanoi (200 mL) was added diethyl 2-(ethoxymethy!ene)malonate (45.7 mL, 228 mmol), and the resluiting mixture was heated at 80 °C for 1 h. LC S indicated completion of the reaction. The mixture was concentrated in vacuo to remove ethanoi and then washed with hexane to afford diethyl 2-(((2-fiuoro-3,4-dimethoxyphenyi)amino)methylene)malonate (72 g, 21 1 mmol, 93 % yield) as a brown solid. LCMS (M+H)⁺: 342.1. Example 1 Se Ethyl 8-fiuoro~6,7~dimethoxy-4-oxO-1 ,4-dihydroquinollne-3~carboxylate

To Dowtherm (25 mL) heated at 250 "C was added diethyl 2~(((2~fiuoro~3,4~

dimethoxyphenyl)amino)methylene)malonate (10 g, 29.3 mmol). The mixture was stirred at the same temperature for 40 mins, LCMS indicated completion of the reaction. The reaction was allowed to cooi at rt for 5 mins and was then poured into cold hexane, the yellow precipitate was collected by filtration, washed with hexane and dried In the air. This reaction was carried out in similar scale for several times. From a total of 72 g of diethyl 2-(((2-fluoro-3,4-dimethoxyphenyl)amino)methylene)malonate (21 1 mmol), ethyl 8-fluoro- 6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxyiate (38 g, 129 mmol, 81.0 % yield) was obtained as a yellow solid. LCMS (M+H)⁺: 296.0

E am|^le_n15f Ethyl 1-ethyl-8-fluoro-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3- carboxyiate PU85023

A mixture of ethyl 8-fiuoro-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoi!ne-3-carboxylate (41 g, 139 mmoi) and potassium carbonate (28.8 g, 208 mmo!) in triethy! phosphate (1 18 ml, 694 mmoi) was stirred at 120 °C for 5h. LCMS indicated completion of the reaction. Then the reaction mixture was cooled down and diluted with water, extracted with DC , dried over sodium sulfate and evaporated in vacuo. The crude material was purified by reverse phase automatic C-18 column chromatography (Combif!ash RF, 130g column), eluting with Acetortitriie/ Water (10-80%) to afford ethyl 1-ethyl-8-fluoro-6,7-dimethoxy-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate (26 g, 80 mmoi, 57.9 % yield) as a white solid.

LCMS (M+H)⁺: 324.1.

Exampte 15q 1~Ethyi-8~fiuoro~6,7~dihydroxy~4-oxo~1 ,4~dlhydroqulnoline~3-carboxylic acid

To a solution of ethyl 1 -ethyl-8-fluoro-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3- carboxyiate (24 g, 74.2 mmoi) in Dlchioromethane (DCM) (300 mL) was added boron tribromide (35.1 mL, 371 mmoi) at -78 "C. The mixture was warmed up to rt and stirred at 25 °C overnight. LCMS indicated completion of the reaction. The mixture was diluted with EtOH and concentrated in vacuo. The same procedure was repeated several times to afford 1-ethyl-8-fiuoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolir!e-3-carboxylic acid (18 g, 67.4 mmoi, 91 % yield) as a yellow solid. This product was used in next step reaction without further purification. LCMS (M+H)⁺: 268.0. Exarnpfe 15 4-Methoxybenzyl 1-ethyl-8-fiuoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate PU85023

To a solution of 1-ethyl-8-fluoro-6,7-dihydroxy-4-oxo-1.4-dihydroquinoline-3-carboxylic acid (18 g, 67.4 mmol) in N.N-Dimethyiformamide (D F) (200 mL) was added K₂C0₃ (48.5 g, 337 mmoi), followed by 1-(chloromethyl)-4-meihoxybenzene (36.8 mL, 289 rnmo!). The reaction mixture was stirred at 50 °C overnight. LCMS indicated completion of the reaction. Water (500ml) was added and the mixture was stirred at r.t for 15mins. The precipitate was collected by filtration and washed with water to afford 4- methoxybenzyl 1-ethyi-8-fiuoro-6_!7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate (32 g, 51.0 mmoi, 76 % yield) as a yellow solid.

LCMS (M+H)⁺: 628.4.

Exampte 15Ϊ 1-Ethyl-8-fluoro-6_!7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline- 3-carboxylic acid

To a suspension of ethyl 1-ethyl-8-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- di ydroquinoiine-3-carboxylate (32 g, 59.8 mmol) in a mixture of Methanol (250 mL) and Water (125 mL) was added KOH (8.70 g, 120 mmol) portionwise. The resulting mixture was stirred at 90 "C for 3 h. LCMS Indicated completion of the reaction. The mixture was cooled down to r.t, and concentrated. Water was then added, and the mixture was adjusted to pH -1 using 8 N aq. HCI. The precipitate was collected by filtration and dried to afford 1-et yl-8-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxyllc acid (28 g, 55.2 mmol, 92 % yield) as a yellow solid. LCMS ( +H)⁺: 508.2.

Exampte 15Ϊ 1-Ethyl-8-fluoro-3-(hydroxymethyl)-6,7-bis((4-methoxybenzyl)oxy)quinolin- 4(1 H)-one PU85023

To a suspension of 1-et yl-8-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroqulno!ine-3-carboxyilc acid (15 g, 29.6 mmoi) in Tetrahydrofuran (THF) (100 mL) was added TEA (8.18 mL, 44.3 mmoi) and isobutyl eh!oroformate (5.82 mL, 44.3 mmoi). The resulting mixiure was stirred at r.t. for 1 h. The mixture was then cooied down to -40 °C and a solution of DIBAL-H (49.3 mL, 73.9 mmoi) in toluene (1.5 ) was added. The mixture was stirred at the same temperature for 1 .5 h, when LC S indicated completion of the reaction. The reaction was quenched with sat. NH₄Ci (aq.), warmed up to r.t. and concentrated. The mixture was diluted with sat. NH₄Ci and extracted with

dichioromethane (2 x 30 mL). The combined organic solution was then dried over sodium sulfate, filtered and concentrated in vacuo to afford 1~ethy!~8-fluoro-3~(hydroxymethyi)~ 6,7-bis((4-methoxybenzyi)oxy)quinolin-4(1 H)-one (10 g, 20.26 mmoi, 68.8 % yield) as a yellow solid, which was used in the next oxidation step without further purification.

LCMS (M+H)⁺: 494.2.

Example 15k 1-Ethyl-8-fluoro-6_!7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinolirie- 3-earba!dehyde

To a yellow solution of 1-ethyl-8-fluoro-3-(hydroxymethyl)-6,7-bis((4- methoxybenzyl)oxy)quino!in-4(1 H)-one (10 g, 20.26 mmoi) in Dichioromethane (DCM) (100 mL) was added manganese dioxide (17.62 g, 203 mmoi), and the mixture was stirred at 25 °C for 6.5 h. LCMS indicated completion of the reaction. The solid was filtered off through Ceilte and washed with DCM, the filtrate was concentrated to afford 1 - ethyl-8-fiuoro-6 -bis((4-rnethoxybenzyi)oxy)-4-oxo-1 ,4-dihydroquinoline-3-carbaldehyde (6.5 g, 13.22 mmoi, 85.3 % yield) as a yellow solid that was used in the next step without further purification, LCMS (M+H)⁺: 492.2, PU85023

To a solution of 1-ethyl-8-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carbaldehyde (8.5 g, 13.22 mmoi) in 1.2-Dichloroethane (DCE) (100 mL) was added pyrrolidine (1.637 mL, 19.84 mmoi), sodium triacetoxyborohydride (5.61 g, 26.4 mmoi) and AcOH (0.076 mL, 1.322 mmoi) . The reaction mixture was stirred at 25 °C for 3h. LCMS indicated completion of the reaction. The mixture was diluted with DCM and washed with brine. The organic !ayer was dried over sodium sulfate and evaporated in vacuo. The crude material was purified by normal phase automatic sl!ica column chromatography (Combif!ash RF, 40g golden column) elutlng with eOH/DCM (0-20%), and purified again with reverse phase automatic silica column chromatography

(Combifiash RF, 150g golden column) eluting with acetonitrile/Water (0-60%) to afford 1- ethyl-8-fiuoro-6 -bis((4-methoxybenzyi)oxy)-3-(pyrrolidin-1 -ylmethy!)quinolin-4(1 H)-one (4.3 g, 7.87 mmoi, 59.5 % yield) as a pale yellow solid.

Example 15m (5R,6R,7R)-4-methoxybenzy! 7-((Z)-2-(((1-(tert-butoxy)-2-methy!-1- oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)acetamido)-3- (iodomethyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide

To the solution of (5R,8R,7RV4-methoxybenzyi 7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1 - PU85023 oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetamido)-3- (chloromethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5~oxide (5g, 8.28 mmo!) In Acetone (50 m!_) was added sodium iodide (1.178 g, 7.85 mmol) and the mixture was stirred at rt for 2h. Solvent was removed under vacuum, the residue was chromatographed to give (5R,6R,7R)-4-methoxybenzyi 7-((Z)-2-(((1 -(tert-butoxy)-2- methyl-1-oxopropan-2-y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4- y!)acetarnido)-3-(iodomethyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2-carboxy!ate 5- oxlde (4.5 g, 5.07 mmol, 81 % yield). LCMS: (M+H)+: 887.6

Example 15r¾ 1-(((6R,7R)-7-((Z)-2-(((1-(tert-butoxy)-2-methyi-1 -oxopropan-2-yl)oxy) imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetam!do)-2-(((4- methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yi)rnethyl)-1 -((1- ethyl-8-fiuoro-6 J-bis((4-methoxybenzyi)oxy)-4-oxo-1.4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium

A solution of (5R,6R,7R)-4-methoxybenzyl 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)acetamido)-3- (iodomethyi)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2-Garboxylate 5-oxlde (1.624 g, 1.829 mmol) and 1 -ethyl-8-fluoro-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidiri-1- yimethy!)quinolin-4(1 H)-one (Example 151) (1 g, 1 .829 mmol) in Ν,Ν-dimethylacetamide (DMA) (10 mL) was stirred at room temperature for 3h and was then left overnight in the refrigerator. Ν,Ν-dimethy!formamide (DMF) (10.00 mL) was added and the mixture was cooled to -40°C followed by addition of PBr₃ (0.345 mL, 3.66 mmol). Stirred at -4Q°C for 30 min. Quenched with 5% sodium chloride solution. Filtered. Chromatographed on ISGO silica gel column eluting with 0-20% methanol: dichloromethane to give 1 -(((6R.7R)-7- ((Z)-2-(((1-(tert-butoxy)-2-methyl-1 -oxopropan-2-yi)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)acetamido)-2-(((4-methoxybenzyl)oxy)carbonyi)-8-oxo- PU85023

5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-((1-ethyl-8-fluoro-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinolin-3-yi)methyi)pyrroiidin-1-ium (2.67g, 2.067 mmol, 1 13 % yield). LCMS: (M+H)+: 1291.2 Example 15o (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropar!-2-yl)oxy) imino)acetamido)-3-((1-((1-ethyl-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2^ carboxylate, Sodium salt

To the solution of 1-(((6R,7R)-7-((Z)-2-(((1-(tert-butoxy)-2-methyi-1 -oxopropan-2- yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-2-(((4- methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yi)rnethyl)-1 -((1- ethyl-8-fiuoro-6.7-bis((4-methoxybenzyi)oxy)-4-oxo-1.4-dihydroquinolin-3- yl)methyl)pyrrolidin-1 -ium (2.6g, 2.013 mmol) in dichloromethane (DGM) (30 mL} was added anisole (2.199 mL, 20.13 mmol) followed by trifiuoroacertic acid (4.85 mL, 80.4 mmol) at 0"C. The mixture was allowed to warm up to room temperature and stirred at room temperature for 18h. Solvent was removed and residue triturated with isopropyl ether. Filtered, Dissolved in acetonitrile, water, 2N hydrochloric acid and HP~20-ss resin added. Solvent removed. Purified through HP-20-ss resin and C18 ISCO column. Desired fractions were stripped and Iyopnilized to constant weight. 1 eq of 0.2N sodium hydroxide was added, followed by dry ice. Lyophilized to give (6R,7R)-7-((Z)-2-(2-aminothiazoi-4- yi)-2-(((2-carboxypropan-2-yl)oxy)imino)acetamido)-3-((1-((1-ethyi-8-f!uoro-6,7-dihydroxy- 4-oxo-1 ,4-dihydroquinolin-3-y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-th azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Sodium salt (442 mg, 0.555 mmol, 27.6 % yield). LCMS: (M+H)+: 774.3; 1 H NMR (400 MHz, DEUTERIUM OX!DE) δ ppm 1.31 - 1.45 (m, 9 H) 2.13 (br. s., 4 H) 3.58 (br. s.,6 H) 3.75 - 4.16 (m, 4 H) 4.49 (d, J-7.58 Hz, 4 H) 5.45 (d,J=5.Q5 Hz, 1 H) 5.80 (s, 1 H) 6.93 (s, 1 H) 7.35 (s, 1 H), 8.15 (s, 1 H).

■Exam le 16 (6R_!7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino) PU85023 acetamido)-3-((1-((5-chloro-1 -ethyl-67-dihydroxy-4-oxo-1 ,4-dihydroquinoli

y!)methyl)pyrro!idin-1-ium-1-yl)methyi^

carboxy!ate

Exampje 18a 5-Ethyl-9-nitro-8-oxo-5,8-dihydro-[1 ,3]dioxala[4,5-gjquinoiine-7-carboxylic acid

5-Ethyl-8-oxo-5,8-dihydro-[1 ,3]dioxoio[4,5-g]quinoline-7-carboxylic acid (50 g, 191 mmol) was dissolved in H₂SO,: (1 12 ml, 2105 mmol) with stirring. The thick solution was cooled to 0 °C and potassium nitrate (21.29 g, 21 1 mmol) was added In small portions. The temperature of the reaction mixture was maintained below 10 °C by use of an Iced water bath. After the addition, the mixture was kept under 10 °C for 1 h, and then was allowed to warm up to r.t, and stirred for overnight. It was poured into iced water (2 L). The yellow precipitates were collected by filtration, washed with water and ethanol, and dried in vacuo to afford 5-ethyl-9-nitro-8-oxo-5,8-dihydro-[1 ,3]dioxolo[4,5-g]qulnoline-7-carboxyilc acid (61 g, 187 mmol, 98 % yield) as a light brown solid. LCMS: (M+Hf: 307.4.

Exampte 18b 9-Amino-5-ethyl-8-oxo-5,8-dihydro-[1 ,3]dioxolo[4,5-g]quinoiine-7-carboxylic acid

A suspension of 5-Ethyl-9-nitro-8-oxo-5,8-dihydro-[1 ,3]dioxoio[4,5-g]quino!ine-7~

carboxylic acid (6 g, 19.59 mmol) and Pd/C (1 g, 0.940 mmol) in a mixture of acetic acid (150 mL) and con. HCi (50 mL) was hydrogenated on a Parr apparatus at 50 psi of H₂ for 1 h at r.t. LCMS indicated completion of the reaction. The catalyst was filtered off. The PU85023 filtrate was added dropwise into water (675 mL). The light yellow precipitates were collected by filtration, washed with water and dried in vacuo to afford 9-amino-5-ethyl-8- oxO-5,8-dihydro-[1 ,3]dioxoio[4,5-g]quinoline-7-carboxylic acid (4.72 g, 17.09 mmol, 87 % yield). LCMS: (M+Hf: 277.2.

Example 18c 7-Carboxy-5-ethyl~8-oxo-5,8-dihydro-[1 _!3]dloxoio[4,5~g]quinoiine- diazonium chloride

To a pale brown suspension of 9-amino-5-ethyi-8-oxo-5,8-dihydro-[1 ,3]dioxoio[4,5- g]quino!ine-7-carboxy!ic acid (1 1 g, 39.8 mmol) in con. HCI (25 mL) stirred at r.t. was added dropwise sodium nitrite (3.57 g, 51.8 mmol) In water (9 mL) at such a rate that the temperature of the reaction mixture would not exceed 45 °C (Ice bath! Slow addition! ). Stirring continued for 5 h. The solution was poured into 120 mL of water and allowed to stand overnight in the refrigerator. The precipitate was collected by filtration, washed with water and dried to afford 7-carboxy-5-ethyi-8-oxo-5_>8-dihydro-[1 ,3]dioxoio[4,5-g]quinoline- 9-diazonium, Chloride (5 g, 15.45 mmol, 38.8 % yield). LCMS: (M+H)⁺: 324.2.

Note: From LCMS and NMR, the solid was a mixture of the title compound and 5- chloro~1-ethy!-6,7-dihydroxy-4~oxo-1 ,4-dihydroquino!ine-3~carboxyiic acid (Example 16d).

-Chloro-1-ethyl-6.7-dihydroxy-4-oxo-1.4-dihydroquinoline-3-carboxylic acid

A solution of 7-carboxy-5-ethy!-8-oxo-5,8-dihydro-[1 ^jdioxoio^.S-gjquinoline-Q- diazonlum chloride (7.5 g, 23.17 mmol) (Mixture of 16c and 16d) in 50% H₂S0₄ (60 mL) was heated at 95 °C for 4 h. The reaction mixture was cooled down to r.t. and then poured into 450 mL of water. The precipitates were collected by fiitration and dried to PU85023 afford 5-chioro-1 -ethyl-6 -dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxyiic acid (8 g, 18.81 mmol, 80 % yield) as a yellow brown solid. LCMS: ( ÷Hf: 284.1.

Example 18e 4-Methoxybenzy! 5-chioro-1-ethyi-6,7-bis((4-methoxybenzyl)oxy)-4-oxo- 1 ,4-dihydroquinoiine-3-carboxylate

To a red suspension of S-chloro-l-eihyi-GJ-dihydroxy^-oxo-l ^-dihydroquinoiiRe-S- carboxyilc acid (12.2 g, 43.0 mmol), K_zC0₃ (23.78 g, 172 mmol) and ΚΪ (0.714 g, 4.30 mmol) in Ν,Ν-Dimethyiformamide (DMF) (200 mL) at r.t. was added 1-(chloromethyl)-4- methoxybenzene (19.28 mL, 142 mmol). The mixture was heated at 90 °C for 4 h. After cooling, the mixture was concentrated and partitioned between water (200 mL) and DCM (200 mL), and extracted with DCM (150 mL) twice. The organic layers were combined, dried and concentrated. The crude product was purified twice through column chromatography eluting with EtOAc/hexanes (0-100%) to afford pure 4~methoxybenzyi 5- chioro-1-ethyi-6,7-bis((4-methoxybenzy!)oxy)-4-oxo-1 ,4-diriydroquinoiine-3-carboxyiate (14 g, 20.85 mmol, 48.0 % yield) as a white solid. LC S: (M+H)⁺: 644,1 . Exampte 18f 5-Chloro-1 -ethyi-6,7-bis((4-methoxybenzy!)oxy)-4-oxo-1 ,4-dihydroquinoiine- 3-carboxylic acid

To a suspension of 4-methoxybenzyl 5-chioro-1-ethyi-6,7-bis((4-methoxybenzy!)oxy)-4- oxo-1 ,4-dihydroquinoline-3-carboxylate (8.4 g, 13.04 mmol) in a mixture of Methanol (90 mL) and Water (30.0 mL) was added KOH (1.483 g, 28.1 mmol) portionwise. The resulting mixture was stirred under reflux for 1 .5 h. The mixture was then cooled down to r.t., concentrated, diluted with water (150 mL) and adjusted pH to 2-3 using 6 N HCI (aq.). The precipitates were collected by filtration and dried to afford 5-chioro-1-ethyi-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3-carboxylic acid (6.7 g, 12.79 mmol, 98 PU85023

% i yield) as a white solid. LCMS: (M+H)⁺: 524.2.

Exampte 18g 5-ch!oro-1-ethyl-3-(hydroxymethy!)-6 ,7-bis((4-methoxybenzyl)oxy)quinoiin-

To a suspension of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 _>4- dihydroquinoiiRe-3-earboxyiic acid (24 g, 41.7 mmoi) in Tetrahydrofuran (THF) (450 mL) was added triethyiamine (8.71 mL, 82.5 mmoi), followed by careful addition of isobutyl chloroformate (7.86 mL, 58.4 mmoi). The resulting mixture was stirred at r.t. for 2.5 h. Then the mixture was cooled down to - 40 °C and a solution of DIBAL-H (89.5 mL, 104 mmo!) in toluene (1.5 M) was added. The mixture was stirred at the same temperature for 3.5 h. After completion, the reaction was quenched with sat. NH^Ci (aq.), warmed up to r.t., diluted with sat. NH^CI (aq.), filtered through Celite to remove gummy precipitate, and extracted with dichloromethane twice. The combined organic extracts were then saturated with brine and then dried over sodium sulfate, filtered and the resulting filtrate was concentrated in vacuo to afford 5-chloro-1 -ethyl-3-(hydroxymethyl)-6,7-bis((4- methoxybenzyl)oxy)quinoiin-4(1 H)-one (19 g, 30.9 mmo!, 74.2 % yield) as a yellow solid, which was used in the next oxidation step. LCMS: (M+H)⁺: 5 0.0.

Example 16h 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline- 3-carbaldehyde

To a yellow solution of 5-chloro-1 -ethyi-3-(hydroxymethyi)-6,7-bis((4-methoxybenzyl)oxy) quinolin-4(1 H)-one (19 g, 30.9 mmoi) in Dichloromethane (DCM) (250 mL) was added manganese dioxide (41 .1 g, 402 mmoi), and the mixture was stirred at r.t. for overnight. PU85023

The mixture was then filtered through Celite and washed with DCM. The filtrate was concentrated to afford 5-chloro-1 -eihyl-6,7-bls((4-methoxybenzy!)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carba!dehyde (17 g, 27.4 mmoi, 89 % yield) as a yellow solid, which was used in the next step without further purification. LCMS: ( +H)⁺: 508.1.

Example 18i 5-chioro-1-ethyi-6,7-bis((4-methoxybenzyi)oxy)-3-(pyrroiidin-1 - y!methyi)quinolin-4(1 H)-one

To a solution of 5-chioro-1-ethy!-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydro quinoiine-3-carbaldehyde (21 .3 g, 34.4 mmoi) in 1 ,2-Dichioroethane (DGE) (200 mL) was added pyrrolidine (3.13 mL, 37.8 mmoi) and sodium triacetoxyborohydrlde (10.93 g, 51.6 mmo!). The mixture was stirred at r.t. for 3 h. Then reaction mixture was concentrated, partitioned between DCM and brine, and extracted with DCM twice. The combined organic extracts were dried over sodium sulfate and the solvent was removed in vacuo. The crude product was then purified through normal phase chromatography (CombiFiash Rf), eluting with [DCM/MeOH/NH₄OH (v:v:v = 80:20:2)]:DCM (0% to 60%) to afford 5- chioro-1-ethy!-6,7-bis((4-methoxybenzy!)oxy)-3-(pyrro!idin-1-ylmethyl)quino!in-4(1 H)-one (17 g, 30.2 mmoi, 88 % yield) as a light yellow solid. LCMS: (M+H)⁺: 563.2.

Example 18i (S)-4-tert-butyl 1 -(4-methoxybenzyl) 2-(((Z)-(1 -(2-((tert- butoxycarbonyi)amino)thiazoi-4-y!)-2-(((5R,6R,7R)-3-(iodomethyi)-2-(((4- methoxybenzyi)oxy)carbonyi)-5-oxido-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-en-7- yi)amino)-2-oxoethyiidene)amino)oxy)succinate

PU85023

To a solution of (S)-4-tert-butyl 1 -(4-methoxybenzyl) 2-(((Z)-(1-(2-((tert-butoxycarbonyi) amino)thiazoi-4-y!)-2-(((5R,6R7R)-3-(ch!oromethy!)-2-(((4-met oxybenzyl)oxy)carbonyl)- 5-oxido-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-eri-7-yi)amino)-2-oxoethylidene) amino) oxy)succinate (10.60 g, 1 1 .20 mmol) (from Shionogi) in Acetone (100 mL) at ri was added sodium iodide (2.52 g, 16.80 mmol). The mixture was stirred at the same temperature over 2 h. LCMS indicated completion of the reaction. The solid was filtered off, the filtrate was concentrated under vacuum, and the residue was purified by Combif!ash automated silica gel chromatography (120 g Gold column), e!uting with ethyl acetate/hexanes (0- 50%) to afford (S)-4-tert-butyi 1-(4-methoxybenzyl) 2-(((Z)-(1-(2-((tert-butoxycarbonyl) amino)thiazoi-4-yl)-2-(((5R,6RJR)-3-(iodomethyl)-2-(((4-methoxybenzyl)oxy)carbonyl)-5- oxido-8-oxo-5-thia-1-azabicycioi4.2 ]oct-2-en-7-yl)amino)-2-oxoethyiidene)amino) oxy) succinate (9.7 g, 9.35 mmol, 83 % yield) as a yellow solid. LCMS: (M+H)⁺: 1037.8. Exampte 18k 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-meihoxybenzy )oxy)-1 ,4- dioxobutan-2-y!)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetami (((4-met oxybenzy!)oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2 ]oct-2-en-3-yl)methyi)-1 - ((5-chloiO-1-ethyl~6,7~bls((4~methoxybenzyl)oxy)-4~oxo~i ,4~dihydroquinolin~3- y!)rnethyl)pyrro!idin-1-ium

To a suspension of 5-chloro-1-ethyl-6 ,7-bis((4~methoxybenzyl)oxy)-3~(pyrralldin-1 - ylmethyl)quinolin-4(1 H)-one (Example 16i) (2.0 g, 3.55 mmol) in DMA (20 mL) at 15°C under N2 was added (S)-4-tert-butyl 1-(4-methoxybenzyl) 2-(((Z)-(1-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)-2-(((5R,6RJR)-3-(iodomethyl)-2-(((4-methoxy benzyi)oxy)carbonyl)-5-oxido-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-7-yi)amino)-2- oxoethylidene)amino)oxy)succinate (3.89 g, 3.55 mmol) In DMA (8 mL). The mixture became clear solution after a few minutes and stirring continued at the same temperature over 2 h, Ν,Ν-Dimethylformamide (DMF) (58.00 mL) was then added. The mixture was cooled to -7Q°C and treated with PBr₃ (0.737 mL, 7.81 mmol) dropwise. The mixture was allowed to warm up to -40°C. LCMS indicated complete reduction of sulfoxide. The organic solution was poured into ice-cooled 5% NaCI (250 mL, containing 0.05M HC!) and stirred for -15 mln. The solid was separated by filtration, washed with water and dried under high vacuum to afford 1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4- methoxybenzyl)oxy)-1 ,4-dioxobutan-2-yi)oxy)imino)-2-(2-((tert-butoxycarbonyl)amirio) thiazol-4~yi)acetamido)~2-(((4-methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1~azabicycio

[4.2.0]oct-2-en-3-yl)methyi)-1-((5-chioro-1 -ethyi-6J-bis((4-methoxybenzy!)oxy)-4-oxo-1 ,4- dihydroquinoiin-3-yl)methyi)pyrrolidin-1-ium (5.99 g, 84% purity, 3.45 mmoi, 97 % yield) as a brown solid. LCMS: (M+H)⁺: 1458.5. Exarnpfe 188 (6R.7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-d!hydroxy-4-oxo-1 ,4-d!hydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate

To a solution of 1 -(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1 -((4-methoxybenzyl)oxy)-1 .4- dioxobutan-2-yi)oxy)imino)-2-(2-((tert-bLitoxycarbonyi)amino)thiazol-4-yi)acetamido)-2- (((4-methoxybenzyi)oxy)carbonyl)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yl)methyi)-1 - ((5-chloro-1-ethyi-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-d!hydroquinoiin-3- y!)methyl)pyrro!idin-1 -ium (5.99 g, 3.45 mmol) in Dichioromethane (DCM) (30 mL) under N2 at 0°C was added anisole (3.77 mL, 34.5 mmol), followed by 2,2,2-triliuoroacetic acid (10.0 mL, 130 mmoi). The mixture was warmed up to ri and stirred at rt over night. LCMS

Indicated completion of the deprotection. The mixture was cooied to 0°C, Diisopropyi ether (100 mL) was then added. The mixture was stirred for 10 min and the solid was collected by filtration, and washed twice with diisopropyi ether (2x20 mL). The solid (-4.4 g) was dissolved in a mixture of CH3CN (25 mL)/water (25 mL)/2M HCI (10 mL), HP20SS resin (30 g) was then added. The mixture was concentrated to dryness, and the resin was loaded in a pre-column containing HP20SS resin (45 g), and was then purified by reverse phase Combiflash using C18 column and eluting with 0-30% CI-^CN/water to afford (8R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -et yl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoliri-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate (F1 : 0,975 g, 1.189 mmoi, 34.4%) as a slightly brown solid. LCMS: (M+H)⁺: 820.2. Ή NMR (400 MHz, METHANOL-cW) δ ppm 1 .50 (t, J=7.07 Hz, 3 H) 2.22 (br. s., 4 H) 2.93 - 2.99 (m, 2 H) 3.44 - 3.65 (m, 6 H) 3.89 - 3.98 (m, 1 H) 4.09 - 4.21 (m, 1 H) 4.27 - 4.41 (m, 3 H) 4.46 - 4.55 (m, 1 H) 5.09 - 5.16 (m, 1 H) 5.25 - 5.33 (m, 1 H) 5.85 - 5.90 (m, 1 H) 6.93 (s, 1 H) 7.04 - 7.1 1 (m, 1 H) 8.19 - 8.26 (m, 1 H)

Exarnpfe 17 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-d!hydroxy-4-oxo-1 ,4-d!hydroquinolin-3-yl) methyl)pyrrolidin-1-ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, Sodium salt

(6R7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino)acetamido)-3- ((1-((5-chioro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiln-3-y!)methyl)pyrro!idln-1- lum-1 -yl)meihyl)-8-oxo-5-thia-1 -azablcyclo[4.2.0joct-2-ene-2-carboxylate (Example 161) (0.233 g, 0.284 mmoi) was suspended in pure water (HPLC grade, 15 mL) with the help of sonication, and cooied to 0°C. With vigorous stirring, aq NaOH (0.2 N, 1.42 mL, 0.284 mmoi) was added slowly into the suspension from an Eppendorf Pipette. After the addition a small piece of dry ice was added to quench any extra NaOH. The pale yellow solution was then frozen and iyophilized to afford (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yi)- 2~(((S)-1 ,2-dicarboxyethQxy)lmino)acetamido)-3-((1 -((5-chloro-1 -ethyl-6,7-dihyrJroxy-4- PU85023

QXG-1 ,4-dihydroquinolin-3-yl)methyi)pyrroiidin-1-ium-1 -yl)methyl)-8-oxo-5-t ia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Sodium salt (0.239 g, 0.278 mmo!, 8.05 % yield) as an off-white solid. LCMS: ( +H)⁺: 820.3. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.30 (t, J=7 Λ7 Hz, 3 H) 2.01 (br. s„ 4 H) 2.59 - 2.68 (m, 2

H) 3, 18 - 3.47 (m, 6 H) 3.87 (d, J=16.67 Hz, 1 H) 3.94 - 4.36 (m, 5 H) 4,78 - 4.88 (m, 1 H) 5.28 (d, J-5.05 Hz, 1 H) 5.72 (d, J=5.05 Hz, 1 H) 6.88 (s, 2 H) 7.98 - 8.09 (m, 1 H).

Example 18 (6R_>7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imlno) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yl) methyl )pyrroiidin-1-ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiaie, 2 Sodium salt

A suspension of (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1 -((5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate (Example 161) (0.601 g, 0.733 mmol) was cooled to -5-0°C using an acetone-dry ice bath. With vigorous stirring, aq NaOH (0.2 N) was added slowly Into the suspension from an Eppendorf Pipette unti! pH of the mixture reached 6.5 (-7.2 mL of NaOH added). After the addition a small piece of dry ice was added to quench any extra NaOH. The solution was then frozen and !yophiilzed to afford (6R,7R)~7-((Z)-2-(2~ aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chioro-1-ethyi- 6,7-dih droxy-4-oxo-1 ,4-dihydroquinolin-3-yi) methyi)pyrrolidin-1-ium-1 -yi)methyl)-8-oxo- 5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate, 2 Sodium salt (0.633 g, 0.687 mmol, 94 % yield) as an off-white solid. LCMS: (M+H)⁺: 820.3. Ή NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.33 (t, J-6.69 Hz, 3 H) 2.02 (br. s., 4 H) 2.60 (s, 2 H) 3.22 - 3.50 (m, 6 H) 3.82 - 3.93 (m, 1 H) 3.96 - 4.07 (m, 1 H) 4.10 - 4.40 (m, 4 H) 4.81 - 4.90 (m, 1 H) 5.26 - 5.32 (m, 1 H) 5.70 - 5.78 (m, 1 H) 6.90 (s, 2 H) 8.01 - 8.12 (m, 1 H). Exarnpfe 19 (6R_>7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)lmino)acetamido)-3-((1-((5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 _>4-dihydroquinolin-

3-yl)methyi)pyrrolidin-1-ium-1 -yi)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2- carboxyiate PU85023

77--((((ZZ))--22--((((((11--((tteertrt--bbuuttooxxyy))--22--mmeetthhyy!i--11--ooxxoopprrooppaarni--

2-yl)oxy)imino

oxo-5-thia-1 -a

To a solution of (6R,7R)-4-methoxybenzyl 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1- oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-3- (chloromethyl)-8-oxo~5-thia~1-azabicyclo[4.2.0]oct-2~ene-2-carboxylate 5-oxide (10.00 g, 12.56 mmol) in Acetone (100 mL) at rt was added sodium iodide (2.82 g, 18.84 mmol). The mixture was stirred at the same temperature over 2 h. LCMS indicated completion of the reaction. The solid was filtered off, the filtrate was concentrated under vacuum, and the residue was purified by Combiflash automated silica gel chromatography (120 g Gold column), eluting with ethyl acetate/hexanes (0-50%) to afford (6R,7R)-4-methoxybenzyi 7-((Z)-2-(((1-(tert-butoxy)-2-rnethyl-1 -oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxy carbony!) amino)thiazoi-4-yi)acetamido)-3-(iodomethyl)-8-oxo-5-thia-1-azabicyclo[4.2.03 oct-2-ene-2-carboxylate 5-oxide (10.86 g, 12.23 mmol, 97 % yield) as a yellow solid. LCMS: (M+H)⁺: 887.9.

Exampte 19b 1-(((6R,7R)-7-((Z)-2-(((1-(tert-butoxy)-2-methyi-1-oxopropan-2-yl)oxy) lmino)~2-(2~((tert-butQxycarbonyl)amino)thiazoi-4-yl)acetamido)-2-(((4~methoxybenzyi) oxy) carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-((5-chioro-1-ethyi- 6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi)pyrrolidi PU85023

To a suspension of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyi)oxy)-3-(pyrrolidin-1 - y!methy!)quinoiin-4(1 H)-one (Example 16i) (0.982 g, 1.744 mmoi) in DMA (10 mL) under

N₂ at 10°C was added (6R,7R)-4-methoxybenzy! 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1- oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acetamido)-3- (iodomethyl)-8-oxo-5-thia~1 ~azabicyclo[4.2.0]oct-2-ene-2-Garboxyiate 5-oxlde (1.548 g, 1.744 mmol) in DMA (5 mL). The mixture became clear solution after a few minutes and stirring continued at 10-15°C over 2.5 h. Ν,Ν-Dimethylformamide (DMF) (30.00 mL) was then added. The mixture was cooled to ~70°C and treated with tribromophosphine (0.362 mL, 3.84 mmol) dropwise. The mixture was allowed to warm up to -40°C over 30 min. LCMS Indicated complete reduction of sulfoxide. The organic solution was poured into Ice-cooled 5% NaCi (150 mL, containing 0.05M HCI) and stirred for -15 min. The solid was separated by filtration, washed with water and dried under high vacuum to afford 1- (((6R7R)-7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1-oxopropan-2-y!)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)acetamido)-2-(((4-methoxybenzy!)oxy)carbonyi)-8-oxo- 5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1 -((5-chloro-1-ethyi-6.7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinolin-3-yi)methyl)pyrroiidin-1-ium (2.688 g, 80% purity, 1.632 mmol, 94 % yield) as a dark brown foamy solid. LCMS: (M+H)⁺: 1306.1. Example 19c (6RJR)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yi)oxy)irnino) acetamido)-3-((1-((5-chloro-1 -ethyl-6 -dihy

methyl)pyrrolidin-1-ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate PU85023

To a solution of 1 -(((6R,7R)-7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1-oxopropan-2-yl)oxy) imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)acetamido)-2-(((4-methoxyben oxy)carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yl)methyi)-1-((5-chioro-1 -ethyi- 6 -bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi)pyrrolidin-1-ium

(2.868 g, 80% purity, 1.832 mmoi) in Dichloromethane (DCM) (15 mL} under N₂ at 0°C was added anisole (1.783 mL, 18.32 mmoi), followed by TFA (5.0 mL, 64.9 mmo!). The mixture was warmed up to rt and stirred at rt over night. LCMS indicated completion of the deprotection. The mixture was cooled to 0°C. Diisopropyl ether (50 mL) was then added. The mixture was stirred for 10 min and the solid was collected by filtration, and washed twice with diisopropyl ether (2x10 mL). The solid (-2.0 g) was dissolved In a mixture of GH3CN (10 mL)/water (10 mL)/2M HCI (2.5 mL), HP20SS resin (12 g) was then added. The mixture was concentrated to dryness, and the resin was loaded In a pre-column containing HP20SS resin (15 g), and was then purified by reverse phase Comblflash using C18 column and eluting with 0-30% CH₃CN/water to afford (6R,7R)-7-

((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yi)oxy)imino)acetamido)-3-((1 -((5- ch!oro-1-ethy!-6,7-dihydroxy-4-oxo-1 ,4-dlhydroquinolin-3-yi)methyl)pyrroildin~1-lum-1- y!)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate (0.88 g, 0.793 mmo!, 48.6 % yield) as a slightly brown solid. LCMS: (M+H)⁺: 790.1 . Ή N R (400 MHz,

METHANOL-cW) δ ppm 1.50 (s, 3 H) 1.82 (d, 6.57 Hz, 6 H) 2.12 - 2.28 (m, 4 H) 3.42 - 3,63 (m, 8 H) 3.95 - 4,05 (m, 1 H) 4,07 - 4.16 (m, 1 H) 4.26 - 4.40 (m, 3 H) 4.47 - 4.55 (m, 1 H) 5.30 - 5.37 (m, 1 H) 5.89 - 5.97 (m, 1 H) 8.91 (s, 1 H) 7,04 - 7.10 (m, 1 H) 8,17 - 8.24 (m, 1 H). Example 20 (6R_>7R)-7-((Z)-2-(2-aminothiazo!-4-yl)-2-(((2-carboxylatopropan-2-yi)oxy) imlno)acetamldo)-3~((1 ~((5-chioro-1~ethyl-6,7~dihydroxy~4-oxo-1 ,4~dihydroqulnolin~3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en carboxyiate, Sodium salt PU85023

To a suspension of (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yi)-2-(((2-carboxypropan-2-yi)oxy) imino)acetamido)-3-((1 -((5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate (Example 19c) (0,361 g, 0.457 mmol) in Water (20 mL) at 0°C was added 0.2 N sodium hydroxide (2.284 mL, 0.457 mmol) dropwise, followed by a small cube of dry ice. The mixture was then frozen and lyophilized to afford (6R,7R)-7-((Z)-2-(2-amino thiazoi-4-yl)-2-(((2-carboxylatopropan-2-yl)oxy)imino)acetarnido)-3-((1-((5-chloro-1 -ethyl- 6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yl)methyl)pyrrolidin-1 -!um-1-yl)methyl)-8-oxo-5- thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxylate. Sodium salt (0.371 g, 0.41 1 mmol, 90 % yield) as a light brown solid. LC S: (M+H)⁺: 790.1 . H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.20 - 1.48 (m, 9 H) 2.03 (br. s., 4 H) 3.18 - 3.52 (m, 8 H) 3.81 - 4.09 (m, 4 H) 4. 1 - 4.34 (in, 2 H) 5.30 (d, J=5.05 Hz, 1 H) 5.76 (d, J=4.80 Hz, 1 H) 6.69 (s, 1 H) 6.82 (s, 1 H) 7.94 (br. s., 1 H).

Example 21 (6R,7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1 -((5-chloro-1-ethyi-6_>7-dihydroxy-4~oxo-1 ,4-dihydroquinoiin-3~ y!)methyl)pyrro!idin-1~ium-1~yl)methyl)-8-oxo-5-thia-1~azabicyclo[4.2.0joci-2-ene~2- carboxylate, Sodium salt.

To a suspeniion of ethyl 2-(2-aminothiazol-4-yl)-2-oxoacetate (30 g, 150 mmol) in !θ: 3

Dichloromeinane (DCM) (500 mL) was added (Boc)₂0 (48.7 mL, 210 mmo!) and DABCO (18.81 g, 150 mmo!). The mixture was stirred at rt overnight. LCMS indicated completion of the reaction. Solvent was removed under vacuum, and to the resulting residue was added a pre-cooled solution of 2M NaOH (300 mL, 599 mmol) and the mixture was stirred in Ice bath for 2 h. Iced Water (250 mL) was added, and the mixture was carefully acidified with 8N HC! (aq) to pH = 1 , The precipitated material was collected by filtration, washed with water, and dried in vacuo to afford 2-(2-((tert-butoxycarbonyl)amino)thiazol- 4-y!)-2-oxoacetic acid (33.5 g, 123 mmol, 82 % yield) as pale yelow solid. LCMS:

( +Naf : 295.0

Exarnpfe 21 b 2-(2-((tert-butoxycarbonyi)amino)-5-chlorothiazol-4-yl)-2-oxoacetic acid

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(((((RRRRR)))))-----22222,,,,,55555-----dddddiiiiioooooxxxxxooooottttteeeeetttttrrrrraaaaahhhhhyyyyy 22222,,,,,22222,,,,,22222-----tttttrrrrriiiiifffff!lllluuuuuooooorrrrroooooaaaaaccccceeeeetttttaaaaattttteeeee

To neat (R)-2-hydroxysuccinic acid (80 g, 447 mmol) being sirred in ice bath was added TFAA (145 ml. 1029 mmol). The mixture was stirred for 1 h and then at room temperature for 3 h. Toluene was added and the mixture was concentrated under vacuum to afford (R)-2,5-dioxotetrahydrofuran-3-yl 2,2,2-trifluoroacetate (94.7 g, 447 mmol, 100 % yield) PU85023 as a white solid which was used for the next step without further purification.

(400 MHz, D SO-d6) δ ppm 3.35 - 3.59 (m, 2 H) 6.10 (dd, 1 H)

Example 21 d (E)-tert-butyl N,N'-diisopropy!carbamirnidate

To a solution of N,N'-methanediy!ideneh!s(propan-2-amine) (307 ml, 1981 mmoi) in t- BuOH (227 ml, 2377 mmoi) at r.t. was added copper(l) chloride (1.961 g, 19.81 mmoi). The reaction was stirred at room temperature for 24 h and the resulting mixture was distilled under reduced pressure to afford (E)-tert-buty! N,N'-diisopropylcarbamimidate (279 g, 1393 mmoi, 70.3 % yield) as a colorless liquid. 'H NMR (400 MHz,

CHLOROFORM-cM) δ ppm 1 .04 (d, J-6.06 Hz, 6 H) 1.08 (d, J-6.57 Hz, 6 H) 1.46 (s, 9 H) 2.98 - 3.19 (m, 1 H) 3.25 (m, 1 H) 3.64 (s, 1 H)

Example 21 e (R)-4-tert-butyl 1-(4-methoxybenzy!) 2-hydroxysuccinate

To a solution of (R)-2,5-dioxotetrahydrofuran-3-yl 2,2,2-trifluoroacetate (85 g, 401 mmoi) in Dichioromethane (DCM) (600 mL) was added (4-methoxypheny!)methanoi (150 mL, 1202 mmoi). The mixture was stirred at rt for 3 h and then extracted three times with 10% aqueous solution of NaHC0₃. The combined aqueous extracts were washed with DCM, acidified to ρΗ 2 with 2 N HCL and then extracted with DCM. The organic layer was dried over sodium sulfate and filtered. To the filtrate was added (E)-tert-butyl Ν,Ν'- diisopropylcarbamimidate (161 g, 802 mmoi) dropwise over 1 h. The mixture was stirred at rt overnight. The precipitate was removed by filtration and the filtrate was purified by normal phase automatic silica column chromatography (Combifiash RF), e!uting with EA Hexane (10-80%) to afford (R)-4-tert-butyi 1 -(4~meihoxybenzy!) 2-hydroxysuccinate (62.5 g, 201 mmoi, 50.2 % yield) as a colorless oil. LCMS: (M+Na)⁺: 333.1

Exampte 21 f (S)-4-tert-butyl 1-(4-methoxybenzy!) 2-((1 ,3-dioxoisoindolin-2-yi)oxy)succinate PU85023

To a mixture of (R)-4-tert-butyl 1-(4-methoxybenzyl) 2-hydroxysuccinate (61 g, 197 mmo!), N-Hydroxyphthalimide (38.5 g, 238 mmo!) and triphenylphosphine (61.9 g, 238 mmo!) in Tetrahydrofuran (THF) (1000 mi_) was added DIAD (45.9 mL, 236 mmoi) dropwise at 0 °C over 30 min. The mixture was stirred at room temperature for 1 h. The solvent was removed under vacuum, and the residue was purified by normal phase automatic silica column chromatography (Combiflash RF), eluting with EA Hexane (0- 90%) to afford (S)-4-tert-butyi 1-(4-methoxybenzyl) 2-((1 ,3-dioxoisoindo!in-2-yl)oxy) succinate (88 g, 193 mmol, 98 % yield) as a pale yellow oil. LCMS: ( +Na)⁺: 478.3 fe 21 g (S)-4-tert-butyl 1-(4-metr hoxybenzy!) 2-(aminooxy)succinate

To a solution of (S)-4-tert-butyl 1-(4-methoxybenzyl) 2-((1 ,3~dioxoisoindoiin-2~ y!)oxy)succinate (88 g, 149 mmol) in Dich!oromethane (DCM) (500 mL) was added methylhydrazine (7.88 mL, 149 mmol) dropwise at 0 ^GC over 30 min. The mixture was allowed to warm to rt and stirred at rt for 1 h. LCMS indicated completion of the reaction. The Insoluble material was removed by filtration and the filtrate was concentrated and diluted with toluene. The resulting precipitate was removed by filtration again and the filtrate was concentrated to afford crude (S)-4-tert-butyl 1-(4-methoxybenzyi) 2- (aminooxy)sLiccinate (47 g, 144 mmol, 97 % yield) as a pale yellow oil. The product was used in next step withour further purification. LCMS: (M+H)⁺: 328.1

Exampje 21 h (S,Z)-2-(((4-(tert-butoxy)-1-((4-methoxybenzyi)oxy)-1 ,4-dioxobutan-2- yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)-5-chlorothiazoi-4-y!)acetic acid PU85023

t-Bu

To a solution of (S)-4-tert-butyl 1-(4-methoxybenzyl) 2-(aminooxy)succinate (39.7 g, 122 mmol) In Methanol (300 mL) was added a solution of 2-(2-((tert-butoxycarbonyl)amino)-5- ch!orothiazol-4-y!)-2-oxoacetic acid (Example 21 b) (34 g, 1 1 1 mmol) at 0 "C. Then the mixture was allowed to warm up to rt and stirred for 2 h. LCMS Indicated completion of the reaction and the product as a mixture of Z/E isomer (19 : 1 ). The reaction mixture was concentrated and the residue was purified by reverse phase automatic C18 column chromatography (Combinfiash RF, 150g column, six runs), eiuting with Acetonitrile/Water (0-90%) to afford (S,Z)-2-(((4-(tert-butoxy)-1 -((4-methoxybenzyl)oxy)-1 _>4-dioxobutan-2- y!)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)-5-chlorothiazoi-4-yl)acetic acid (46 g, 74.9 mmol, 67.6 % yield) as pale yellow solid. LCMS: (M+H)⁺: 614.4. ¹H NMR (400 MHz, METHANOL-cW) δ ppm 1.44 (s, 9 H) 1.55 (s, 9 H) 2.74 - 2.96 (m, 2 H) 3.78 (s, 3 H) 5.06 - 5.13 (m, 1 H) 5.17 (s, 2 H) 6.86 (d, J-8.84 Hz, 2 H) 7.32 (d, J-8.59 Hz, 2 H) Exarnpfe 21 i (S)-4-tert-butyl 1-(4-methoxybenzyl) 2-(((Z)-(2-(((6R,7R)-2-((benzhydryi oxy) carbony!)-3-(chloromethyl)-8-oxo-5 hia-1-azabicyc!o[4.2.0]oct-2-en-7-yl)amino)-1-(2- ((tert-butoxycarbonyl)amino)-5-chlorothiazol-4-y!)-2-oxoethylldene)amino)oxy)succinate

To a suspension of (S,Z)-2-(((4-(tert-butoxy)-1-((4-methoxybenzyl)oxy)-1 ,4-dioxobutan-2- yi)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)-5-chlorothiazoi-4-yl)acetic acid (16.7 g, 27.2 mmol) and (6RJR)-benzhydry! 7-amino-3-(chloromethyl)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Hydrochloride (12.89 g, 28.6 mmol) in Dichloromethane (DCM) (210 mL) at -30 °C was added phenyl phosphorodichioridate (4.95 mL, 32.6 mmol), followed by dropwise N-methylmorphoiine (8.97 mL, 82 mmol) over 15 mins. The mixture was stirred at the same temperature over 3 h. LCMS indicated PU85023 completion of the reaction. The reaction was quenched by addition of 10% aq. citric acid (85 mL), the organic solution was separated and washed with 5% ΝθΗΟΟ , brine, and dried (Na2SC> ), filtered, and concentrated. The residue was purified by CombiFiash automated silica ge! chromatography (330 g column, 0-50% EtOAc/Hexanes) to afford (S)-4-tert-butyl 1-(4-methoxybenzyi) 2-(((Z)-(2-(((6R,7R)-2-((benzhydryloxy)carbonyi)-3- (chloromethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-7-yl)amino)-1-(2-((tert- butoxycarbQnyl)amino)-5-chiorQthiazol~4-yl)~2-oxoethylidene)amlno)oxy)succinate (26 g, 25,7 mmoi, 95 % yield) as a iight yellow foaming solid. LCMS: (M+H)⁺: 1010.6 Exaropte 21 j (S)~4-tert~butyi 1-(4-methoxybenzyl) 2-(((Z)-(2-(((5R,6R,7R)-2-((benzhydryl oxy) carbonyi)-3-(ch!oromethy!)-5-oxido-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-7- y!)amino)-1 -(2-((tert-butoxycarbonyl)amino)-5-chiorothiazol-4-yi)-2- oxoethylidene)amino)oxy)succinate

To a solution of (S)-4-tert-butyl 1-(4-methoxybenzyl) 2-(((Z)-(2-(((6R,7R)-2-((benzhydry! oxy)carbony!)-3-(chioromethyl)-8-oxo-5-thia-1-azabicycio[4.2 ]oct-2-en-7-yi)amino)-1-(2- ((tert-butoxycarbonyl)amino)-5-chlorothiazoi-4-y!)-2-oxoethylidene)amino)oxy)succi (27.5 g, 27.2 mmoi} in Dichloromethane (DCM) (1 L) at -40 °C was added dropwise a solution of m-CPBA (6.40 g, 28.8 mmo!) in Dichloromethane (DCM) (200 mL) over 10 min. The mixture was stirred at the same temperature over 30 min. LCMS indicated completion of the reaction. The mixture was treated with 15% aq. ^328203 (200 mL).

The organic solution was separated and washed with 5% NaHCC^ (400 mL), brine (400 mL), dried (Na2SG^>4), filtered, and concentrated (temperature of the water bath should not be too high— apx. 25 °C). The residue was purified by CombiFiash automated silica gel chromatography (220 g column), e!uting with EtOAc/Hexanes (0-60%) to afford (S)-4- tert-butyl 1-(4-methoxybenzyl) 2-(((Z)-(2-(((5R,6R,7R)-2-((benzhydryioxy)carbonyl)-3- (chloromethyl)-5-oxido-8-oxo-5-thia-1-azabicyc!o[4.2.0]oct-2-en-7-yl)amino)-1-(2-((tert- PU85023 butoxycarbonyi)amino)-5-chlorothiazoi-4-yl)-2-oxoethyiidene)amino)oxy)succina (27.5 g, 25.4 mmol, 94 % yield) as a iight yellow foaming solid. LC S: (M+H)⁺: 1026.3

Example 21 k (S)-4-tert-butyl 1 -(4~methoxybenzyi) 2-(((Z)-(2~(((5R,8RJR)-2-((benzhydry! oxy) carbonyi)-3-(iodomethyl)-5-oxido-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-en-7-yi) arnino)-1-(2-((teri~buioxycarbonyl)amino)-5-chlorothiazoi-4-yl)-2- oxoethylidene)amino)oxy)succinate

To a solution of (S)-4-tert-butyl 1-(4-methoxybenzyi) 2-(((Z)-(2-(((5R,6R,7R)-2-

((benzhydi loxy)carbonyl)-3-(c ioromethyi)-5-oxido-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2- en-7-yl)amino)-1 -(2-((tert-butoxycarbonyl)amino)-5-chlorothiazoi-4-yl)-2- oxoethylidene)arnino)oxy)suGcinate (27 g, 24.98 mmol) In Acetonitrile (300 mL) was added sodium iodide (5.62 g, 37.5 mmol). The mixture was stirred at room temperature over 1.5 h. LCMS indicated completion of the reaction. The solid was filtered off, the filtrate was concentrated under vacuum, and the residue was purified by CombiF!ash Rf automated silica gel chromatography (330 g column), eluting with EtOAc/Hexanes (0- 50%) to afford (S)-4-tert-butyi 1-(4-methoxybenzyl) 2-(((Z)-(2-(((5R,6R,7R)-2- ((benzhydi loxy)carbonyl)-3-(iodomethyi)-5-oxido-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- en-7-yl)amino)-1 -(2-((tert-butoxycarbonyl)amino)-5-chlorothiazoi-4-yl)-2- oxoethylidene)amino)oxy)succinate (18 g, 14.97 mmol, 59.9 % yield) as a red-brown solid. LCMS: (M+H)⁺: 1 1 18.1

.Example 211 1-(((6R,7R)-2-((benzhydryloxy)carbonyi)-7-((Z)-2-((((S)-4-(tert-butoxy)-1 -((4- methoxybenzyl)oxy)-1 ,4-dioxobutan-2-yi)oxy)imino)^

chiorothiazol-4-yi)acetamido)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yi)methyl)-1-((5- chioro-1-ethy!-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4~dlhydroquinolin~3- y!)rnethyl)pyrro!idin-1 -ium PU85023

To a solution of 5-c ioro-1 -ethyi-6 ,7-bis((4-methoxybenzyi)oxy)-3-(pyrroiidin-1-ylrnetriy!) quinolin-4(1 H)-one (Example 16i) (0.736 g, 1.307 mmol) in DMA (5 mL) at 15°C under N₂ was added (S)~4-tert~butyi 1-(4-methoxybenzyl) 2-(((Z)-(2-(((5R,6R,7R)-2-((benzhydryl oxy) carbonyi)-3-(iodomethyl)-5-oxido-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-en-7- y!)amino)-1 -(2-((tert-butoxycarbonyl)amino)-5-chiorothiazol-4-yi)-2-oxoethy!idene) amino)oxy)succinate (1.482 g, 1.307 mmol) in DMA (5 mL). The mixture was stirred at the same temperature over 3 h. Ν,Ν-Dimethyiformamide (DMF) (20.00 mL) was then added. The mixture was cooled to -70°C and treated with PBr₃ (0.271 mL, 2.88 mmol) dropwise. The mixture was allowed to warm up to ~40°C. LC S indicated complete reduction of sulfoxide. The organic solution was poured into ice-cooled 5% NaCI (120 mL, containing 0.05M HCi) and stirred for -15 min. The solid was separated by filtration, washed with water (2 x 10 mL) and dried under high vacuum to afford 1 -(((6R,7R)-2- ((benzhydry!oxy)carbonyl)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-methoxybenzy!)oxy)-1 ,4- dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)-5-chlorothiazoi-4- yl)acetamido)-8-oxo-5-thia-1 -azabicyclo[4.2 ]oct-2-en-3-yl)methyi)-1-((5-chioro-1-ethyi- 6 J-bis((4-methoxybenzyl)oxy)-4-oxo-1 ^-dihydroquinoiin-3-yl)methyi)pyrrolidin-1-ium (2.41 g, 1.018 mmol, 65% purity, 78 % yield) as a dark brown solid. LCMS: (M+H)⁺: 1538.1

Example 21 m (8R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2- rJicarboxyethoxy)imino)acetamidQ)-3-((1 -((5-chloro-1-ethyl~6,7~rJihydroxy~4-oxo~1 ,4~ rJihydroqulnQlin~3-yl)methyl)pyrrolidin-1~lum~1 -yl)methyi)-8-oxo~5 hia~1- azabicyclo[4.2.0]oct-2~ene-2-carboxyiate, Sodium salt PU85023

To a solution of 1-(((6R,7R)-2-((benzhydryloxy)carbonyi)-7-((Z)-2-((((S)-4-(tert-butoxy)-1 - ((4-methoxybenzyl)oxy)-1 ,4-dioxobutan-2-y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)amin 5-chlorothiazol-4-yi)acetamido)-8-oxo-5-thia-1 -azabicyclo[4.2 ]oct-2-en-3-yl)methyi)-1- ((5-chloro-1-ethyi-6J-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoiin-3- yi)methyl)pyrroiidin-1 -ium (2.41 g, 1.018 mmol) in Dich!oromethane (DCM) (15 mL) under

H₂ at 0°C was added anisole (1.1 12 mL, 10.18 mmoi), followed by TFA (5.0 mL, 84.9 mmoi). The mixture was warmed up to rt and stirred at rt over night, LCMS indicated completion of the deprotection. The mixture was cooled to 0°C, Dlisopropyl ether (50 mL) was then added. The mixture was stirred for 10 min and the solid was collected by filtration, and washed twice with dlisopropyl ether (2x10 mL). The crude product was dissolved in a mixture of CHgCN (10 mL)/water (10 mL)/2M HCI (2.5 mL), HP20SS resin

(10 g) was then added. The mixture was concentrated to dryness, and the resin was loaded in a pre-column containing HP20SS resin (15 g), and was then purified by reverse phase Combiflash using C18 column and eiuting with 0-30% C^CN/water to afford

(6R,7R)-7-((Z)-2-(2-amino-5-ch!orothiazol-4-y!)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetarnido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-ene-2- carboxylate (0.504 g, 0.59 mmoi, 43.3% yield) as a slightly brown so!id. Portion of this product (0.283 g, 0.331 mmol) was suspended in pure water (HPLC grade, 20 mL) with the help of sonication, and cooled to 0°C. With vigorous stirring, aq NaOH (0.2 N, 1.855 mL, 0.331 mmol) was added slowly into the suspension from an Eppendorf Pipette. After the addition a small piece of dry ice was added to quench any extra NaOH. The mixture was then frozen and lyophilized to afford (6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)- 2-(((S)-1 ,2-dicarboxyethoxy)imino)acetamido)-3-((1 -((5-chloro-1 -ethyi-6.7-dihydroxy-4- QXQ-1 ,4-dihydroquinolin-3-yl)methyi)pyrroiidin-1-ium-1 -yl)methyl)-8-oxo-5-thi

azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Sodium salt (0.291 g, 0.315 mmol, 95.2 % yield) as a slightly brown solid. LCMS: (M+H)⁺: 854.0. H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.30 (t, J=6.95 Hz, 3 H) 2.01 (br. s„ 4 H) 2.69 (d, J=8.57 Hz, 2 H) 3.23 - PU85023

3.47 (m, 8 H) 3.83 - 3.92 (m, 1 H) 3.96 - 4.04 (m, 1 H) 4.07 - 4.17 (m, 2 H) 4.18 - 4.37 (m, 2H) 4.86 (t, J=6.57 Hz, 1 H) 5.27 (d, J=4.80 Hz, 1 H) 5.67 - 5.76 (m, 1 H) 6.83 - 6.91 (m, 1 H) 7.97 - 8.06 (m, 1 H) Example 22 (6R_!7R)-7-((Z)-2-(2-amino-5-c iorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)lmino)aceiamido)-3-((1-((5-chioro-1-eihyi-6,7-dihydroxy-4~Qxo-1 ,4-dih

3-yl)methyi)pyrrolidin-1-ium-1 -yi)methy!)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2- carboxyiate, Sodium salt

Exampte 22a (Z)-2-(((1 -(tert-butoxy)-2-methy!-1-oxopropan-2-yi)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-y!)acetic acid

To a solution of (Z)-2-(2-aminothiazoi-4-yl)-2-(((1 -(tert-butoxy)-2-rnethyl-1-oxopropan-2- yl)oxy)imino)acetic acid (120 g, 364 mmoi) in Dichioromethane (DCM) (840 mL) in an Ice bath was added TEA (76 mL, 546 mmoi), a solution of Boc-anhydride (1 10 mL, 474 mmoi), and DMAP (8.90 g, 72,9 mmoi). The mixture was stirred at r.t. over night. The solvent was then removed under vacuum. Water (1 L) and diisopropy!ether (700 mL) were added to the mixture. The aqueous layer was separated, washed with toluene and was adjusted to pH ~2 with 2N HCI. The aqueous !ayer was extracted with toluene. The organic layer was concentrated. IPE was added and the Insoluble material was trituiated. The material was collected by filtration and dried to afford (Z)-2-(((1-(tert-butoxy)-2- methyl-1-oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yi)acetic aad (69 g, 145 mmoi, 39.7 % yield) as a white powder. LC S: (M+H)⁺: 430.0

Exam le 22b (Z)-2-(((1-(tert-butoxy)-2-methyl-1-oxopropan-2-yi)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)-5-chlorothiazoi-4-yl)acetic acid PU85023

A solution of (Z)-2-(((1-(tert-butoxy)-2-methyl-1-oxopropan-2-yi)oxy)imino)-2-(2-((tert- butoxycarbonyi)amino)thiazoi-4-yl)acetic acid (70 g, 147 mmoi) and NCS (23.50 g, 176 mmo!) In Ν,Ν-Dimethy!formamide (DMF) (735 mL) was stirred at 40 °C for 2.5 h. The reaction was quenched by addition of (2 L), EtOAc (1 L) and 5% aq. sodium thiosulfate soin (400 mL). The organic layer was separated, and washed with (2 x), brine, dried and evaporated to afford (Z)-2-(((1 -(tert-butoxy)-2-methyl-1-oxopropan-2- y!)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)-5-chlorothiazoi-4-yl)aGetic acid (69 g, 91 % purity, 135 mmoi, 92 % yield) as a light yellow foaming solid. LCMS: ( +H)^÷: 464.2

Example 22c (8R,7R)-benzhydryl 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 -oxopro y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)am

(chloromethyl)-8-oxo~5-thia~1-azabicyclo[4.2.0]oct-2~ene-2-carboxylate

To a suspension of (Z)-2-(((1-(tert-butoxy)-2-methy!~1 -oxopropan-2-yl)oxy)imino)-2~(2- ((tert-butoxycarbonyl)amino)-5-chlorothiazoi-4-y!)acetic acid (20 g, 39.2 mmoi) and (6R,7R)-benzhydryl 7-arnino-3-(cri!oromethy!)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene- 2-carboxyiate, Hydrochloride (18.59 g, 41.2 mmoi) in Dichioromethane (DCM) (300 mL) at -30 °C was added phenyl phosphorodichloridate (7.15 mL, 47.1 mmoi), followed by dropwise 4-methyimorphoi!ne (12.94 mL, 1 18 mmoi) over 15 mins. The mixture was stirred at the same temperature over 3 h. LCMS indicated completion of the reaction. The reaction was quenched by addition of 10% aq. citric acid (120 mL), the organic solution PU85023 was separated and washed with 5% NaHCC^, brine, and dried (^280 , filtered, and concentrated. The residue was purified by CombiFiash automated si!ica gel

chromatography (330 g column, 0-45% EtOAc/Hexanes) to afford (6R,7R)-benzhydry! 7- ((Z)-2-(((1-(tert-butoxy)-2-methyi-1 -oxopropan-2-yi)oxy)irnino)-2-(2-((tert- butoxycarbonyi)amino)-5-chlorothiazo!-4-yl)acetamido)-3-(chloromethyl)-8-oxo-5-th azabicyclo[4.2.0]oct-2-ene-2-carboxylate (29.9 g, 34.7 mmol, 89 % yield) as a light yellow foaming solid, LCMS: ( +H)⁺: 860.3

Example 22d (5R,6R,7R)-benzhydryl 7-((Z)-2-(((1 -(tert-buioxy)-2-meihyl-1-oxopiOpan-2- y!)oxy)imino)~2-(2~((teri-butoxycarbonyi)amino)-5-chlorothiazo^4~y!)acetanido)~3- (chloromethyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2-ene-2-carboxylate 5-oxide

To a solution of (8R,7R)-benzhydry! 7-((Z)-2-(((1-(tert-butoxy)-2-methyi-1 -oxopropan-2- yl)oxy)imlno)-2-(2-((tert-butoxycarbonyl)amino)-5-chlorothiazol-4-y!)acetarnido}-3- (chloromethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxylate (29.9 g, 34.7 mmol) In Dichioromethane (DCM) (1.1 L) at -40 °C was added dropwise a solution of m- CPBA (8.17 g, 36.5 mmol) in Dichioromethane (DCM) (220 mL) over 10 min. The mixture was stirred at the same temperature over 30 min. LCMS indicated completion of the reaction. The mixture was treated with 15% aq. Na₂S₂0₃ (200 mL). The organic phase was separated and washed with 5% NaHC0₃ (400 mL), brine (400 mL), dried (Na₂S04), filtered, and concentrated. The residue was purified by CombiFiash automated silica gel chromatography (330 g column, 0-50% EtOAc/Hexanes) to afford (5R,6R,7R)-benzhydryl 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 -oxopropan-2-y!)oxy)imirio)-2-(2-((tert- butoxycarbonyi)amino)-5-chlorothiazo!-4-yl)acetamido)-3-(chloromethyl)-8-oxo-5-thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide (29 g, 31 .8 mmol, 91 % yield) as a light yellow solid. LCMS: (M+H)⁺: 876.3

Example 22e (5R,6R,7R)-benzhydry! 7-((Z)-2-(((1-(tert-butoxy)-2-methyi-1 -oxopropan-2- y!)oxy)imino)-2-(2-((teri-buioxycarbonyi)amino)-5-chlorothiazoi-4-y!)acetamido)-3- (iodomethyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide PU85023

To a solution of (5R,8R,7R)-benzhydryl 7-((Z)-2-(((1-(tert-butoxy)-2~FTiethy!-1 -oxopropan- 2-yl)Qxy)iiTiino)~2-(2~((tert~butoxycarbonyl)amino)-5-chiorothlazoi-4~yi^

(chlorQmethyl)-8-oxQ-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide (10,5 g, 1 1 .50 mmol) in acetone (120 mL) was added sodium iodide (2.58 g, 17.24 mmoi). The mixture was stirred at room temperature over 1.5 h. LCMS indicated completion of the reaction. The solid was filtered off, the filtrate was concentrated under vacuum, the residue was purified twice by CombiFiash Rf automated silica gel chromatography (120 g column), elutlng with EtOAc/Hexanes (0-45%) to afford (5R,6R,7R)-benzhydry! 7-((Z)-2- (((1-(tert-butoxy)-2-methyl-1-oxopropan-2-yi)oxy)lmino)-2-(2-(fe

5-chiorothiazol-4-yl)acetamido)-3-(lodomethyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene- 2-carboxylate 5-oxide (7.5 g, 7.75 mmol, 67.4 % yield) as a light brown foaming solid. LCMS: (M+H)⁺: 967.9. l-

To a solution of 5-chioro-1 -ethyi-6_>7-bis((4-methoxybenzyi)oxy)-3-(pyrroiidin-1- yimethy!)quinolin-4(1 H)-one (Example 16i) (0.734 g, 1.304 mmol) in DMA (5 mL) at 15°C under N2 was added (8R,7R)-benzhydry! 7-((Z)-2-(((1 -(tert-butoxy)-2-methyi-1- oxopropan-2~yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)-5-chiorothiazol~4- yi)acetamido)-3-(iodomethyl)-8~oxo-5-thia~1-azabicyclo[4.2.0]oct-2~ene-2-carboxyiate 5- oxide (1.282 g, 1.304 mmoi) in DMA (5 mL). The mixture was stirred at the same temperature over 3 h. Ν,Ν-Dimethyiformamide (DMF) (20.00 mL) was then added. The mixture was cooled to -7Q°C and treated with tribromophosphlne (0.270 mL, 2.87 mmol) dropwise. The mixture was allowed to warm up to -40°C. LCMS indicated complete reduction of sulfoxide. The organic solution was poured into Ice-cooled 5% NaCI (120 mL, containing 0.05M HC!) and stirred for -15 mln. The solid was separated by filtration, washed with water (2 x 10 mL) and dried under high vacuum to afford 1 -(((6R,7R)-2- ((benzhydryioxy)carbonyl)-7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1-oxopropan-2- y!)oxy)imino)-2-(2-((tert-butoxycarbonyl)a

thia-1 -azabicyclo[4.2.03oct-2-en-3-yl)methyl)-1 -((5-chloro-1-ethyl-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinolin-3-yi)methy!)pyrroiidin-1 -ium (2.19 g, 1.136 mmol, 87 % yield) as a dark brown solid. LCMS: (M+H)⁺:1388.1 .

Example 22 q (6R,7R)~7-((Z)-2-(2-amino-5-chloi thiazoi-4-y!)-2-(((2-carboxypropan-2- y!)oxy)lmino)aceiamido)-3-((1-((5-chloro-1-ethyl-6_>7-dihydroxy-4~oxo-1 _>4-dihydroquinolin- 3-yl)methyi)pyrrolidin-1-ium-1 -yi)methy!)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2- carboxylate, Sodium salt

To a solution of 1-(((6R,7R)-2-((benzhydryloxy)carbonyi)-7-((Z)-2-(((1-(tert-butoxy)-2- methyl-1-oxopropan-2-yl)oxy)imlno)-2-(2-((tert-butoxycarbonyl)amino)-5-chlorothiazol-4- yl)acetamido)-8-oxo-5 hia-1 -azabicycio[4.2.0]oct-2-en-3-yl)methyl}-1-((5-chloro-1-ethyl- 6,7-bls( 4-methoxybenzyl)oxy)-4-oxo-1 ,4-dlhydroqulnolin-3-yl)methyl)pyrrolidin-1-lum

(2.19 g, 1.136 mmol) in Dichloromethane (DC ) (12 mL) under N2 at 0°G was added anisole (1 .240 mL, 1.38 mmol), followed by TFA (4.0 mL, 51.9 mmol). The mixture was warmed up to rt and stirred at rt over night. LCMS indicated completion of the

deprotection. The mixture was cooled to 0°C, Diisopropy! ether (50 mL) was then added. The mixture was stirred for 10 min and the solid was collected by filtration, and washed twice with diisopropy! ether (2x10 mL). The crude product was dissolved In a mixture of CH₃GN (10 mL)/water (10 mL)/2M HC! (2.5 mL), HP20SS resin (10 g) was then added. The mixture was concentrated to dryness, and the resin was loaded in a pre-co!umn containing HP20SS resin ( 5 g), and was then purified by reverse phase Combiflash PU85023 using C18 column and e!ui!ng with 0-30% CH₃CN/water to afford (6R,7R)-7-((Z)-2-(2- amino-5~chlorathiazal-4~y!)~2-(((2-carb^

ethyi~8,7-dihydroxy-4~oxG-1 ,4-dihydrodnno!m^

oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxy!ate (0.440 g , 0.534 mrnoi , 47.0% yield) as an off-white solid. Portion of this product (0.246 g , 0.298 mrnoi) was suspended in pure water (HPLC grade, 20 mL) with the help of sonication, and cooled to 0°C. With vigorous stirring, aq NaOH (0.2 N, 1 .49 mL, 0.298 mrnoi) was added slowly into the suspension from an Eppendor Pipette. After the addition a small piece of dry ice was added to quench any extra NaOH. The mixture was then frozen and lyophiiized to afford (8R,7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yi)-2-(((2-carboxypropan-2- y!)oxy)lmino)acetamido)-3-((1 -((5-chioro-1 -ethyi-6_>7-dihydroxy-4~oxo-1 _>4-dihydroquinoiin- 3~yl)methyl)pyn lldln-1 -ium-1 -yl)methy!)-8-oxo-5-thia-1 ~azabicyc!o[4.2.0]oct~2-ene~2- carboxylate, Sodium salt (0.253 g, 0.278 mmol, 93.3 % yield) as a pale yellow foamy solid. LCMS: (M+H)⁺:824.2. H N R (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .27 (t, J-8.82 Hz, 3 H) 1 .39 (d , J-6.82 Hz, 6 H) 1 .90 -2.09 (m, 4 H) 3.34 (br. s. , 8 H) 3.84 - 4.1 1 (m, 4 H) 4.15 - 4.36 (m, 2 H) 5.30 (s, 1 H) 5.72 - 5.82 (m, 1 H) 6.69 - 6.80 (m, 1 H) 7.96 (s, 1 H).

Example 23 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropari-2- yl)oxy)imino)acetamido)-3-((1 -((1 -ethyi-6 J-dihydroxy-4-oxo-1 ,4-dihydroqui

yi)methyl)pyrroiidin-1 -ium-1 -yl)methy!)-8-^^

carboxylate, Sodium salt

The compound was prepared according to the procedures In Examples 19b-19c and Example 20, utilizing 1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrroiidin-1 - ylmethyi)quinolin-4(1 H)-one (Example 9a) in place of 5-chioro-1 -ethyi-6.7-bis((4- methoxybenzyl)oxy)-3-(pyrroiidin-1 -yimethyl)quinolin-4(1 H)-one in Example 19b. LCMS: (M+H)⁺: 756.2. Ή NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .24 - 1 .43 (m, 9 H) 2.00 (br. s. , 4 H) 3.29 (br. s. , 6 H) 3.84 - 4.03 (m, 2 H) 4.12 (d , J=7.33 Hz, 2 H) 4.19 - 4.28 (m, 1 H) 4.30 - 4.40 (m, 1 H) 5.29 (d , J=5.05 Hz, 1 H) 5.76 (d, J=4.8Q Hz, 1 H) 6.82 (s, 1 H) 6,86 (s, 1 H) 7.33 (s, 1 H) 8.03 (s, 1 H) PU85023

Exam le 24 (6R JR)-7-((Z)-2-(2-amino-5-ehloro^

yi)oxy)imino)acetamido)-3-((1-((1 -ethyl-6 -dihydroxy-4-oxo-1 ,4-dihydroqui

y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2 carboxyiate

The compound was prepared according to the two-step sequence of Examples 22f-22g, utilizing 1-ethyl-6J-bis((4-methoxybenzyl}oxy}-3-(pyrroiidin-1-ylmethy!)quinolin-4(1 H)-one (Example 9a) in place of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrroiidin-1 - y!methy!)quinolln-4(1 H)-one in Example 22f. LCMS: (M+H)⁺:790.1. H NMR (400 MHz, METHANOL-^) δ ppm 1.52 (s, 3 H) 1 .61 (d, J=1 .52 Hz, 6 H) 2.19 (d, J=9.85 Hz, 4 H) 3,40 - 3.68 (m, 6 H) 3,92 - 4.02 (m, 1 H) 4.07 - 4.17 (m, 1 H) 4.31 - 4.44 (m, 3 H) 4.50 - 4,58 (m, 1 H) 4,98 -5.05 (m, 1 H) 5.25 - 5.32 (in, 1 H) 5.86 - 5.92 (m, 1 H) 7.12 (s, 1 H) 7,66 (s, 1 H) 8.25 (s, 1 H)

Example 25 (6R_>7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetamido)-3-((1-((1 -ethyi-6 J-dihydroxy-4-oxo-1 ,4-di ydroquinoiin y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8^^

carboxyiate, Sodium salt

The compound was prepared according to the procedure from Examples 20, utilizing (6R,7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yi)-2-(((2-carboxypropan-2- y!)oxy)imino)aceiamido)-3-((1-((1 -ethy!-6,7-dihyd! xy-4-oxo-1 ,4-dihyd! quino!in-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate (Example 24} in place of (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2- carboxypropan-2-yl)oxy)imino)acetamido)-3-((1-((5-chloro-1-ethyl-6,7-dihydroxy-4-oxo- 1 ,4-dihydroquinolin-3-yl)methyi}pyriOlldin-1-ium-1 -yl}methyl)-8-oxo-5-thia-1- azabicycio[4.2.0]oct-2-ene-2-carboxylate. LCMS: (M+H)⁺:790.1. H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.32 (s, 3 H) 1 .39 (d, J=6.82 Hz, 6 H) 1 .88 - 2.09 (m, 4 H) PU85023

3.24 - 3.50 (m, 8 H) 3.84 - 3.94 (m, 1 H) 3.94 - 4.04 (m, 1 H) 4.05 - 4.21 (m, 2 H) 4.20 - 4.31 (m, 1 H) 4.31 - 4.43 (m, 1 H) 5.28 (d, J=4.80 Hz, 1 H) 5.77 (d, J=4.80 Hz, 1 H) 6.90 (s, 1 H) 7.38 (s, 1 H) 8.05 (s, 1 H) Example 28 (6R_!7R)-7-((Z)-2-(2-amino-5-c !orothiazol-4-yl)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-3-((1-((1-ethyl-6,7-dihydroxy-4-oxo-1 ,4- dihydroquinoiin-3-yl)methyi)pyrrolidin-1-ium-1 -yi)methy!)-8-oxo-5-t ia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Sodium salt

The compound was prepared according to the two-step sequence of Examples 211-21 m, utilizing 1-ethyl-67-bis((4-met oxybenzyi)oxy)-3-(pyrrolidin-1-ylmethyl)quinolin-4(1 H)-one (Example 9a) in place of 5-chloiO-1-ethyl~8,7~bls((4~methoxybenzyl)oxy)-3-(pyrroiidin-1 - yimethy!)quinolin-4(1 H)-one in Exampie 211. LCMS: ( +Hf: 820.2. Ή NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.33 (t, J=7.07 Hz, 3 H) 2.00 (br. s., 4 H) 2.69 - 2.79 (m, 2H) 3.23 - 3.49 (m, 6 H) 3.82 - 3.93 (m, 1 H) 3.95 - 4.05 (m, 1 H) 4.12 - 4.24 (m, 2 H) 4.25 - 4.42 (m, 2 H) 4.82 - 4.92 (m, 1 H) 4.87 (dd, J=7.83, 5.31 Hz, 2 H) 5.27 (s, 1 H) 5.67 - 5.77 (m, 1 H) 6.96 - 7.06 (m, 1 H) 7.42 (s, 1 H) 8.09 (s, 1 H)

E am le 27 (6R7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ₎2-dicarboxyethoxy)imino) acetamido)-3-((1-(2-(5-chloro-1-ethyl-6.7-dihydroxy-4-oxo-1 .4-dihydroquinoline-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate, sodium salt

Example 27a 5-C !oro-1 -ethy!-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1- y!)ethyl)-1 ,4-dihydroquinoline-3-carboxamide PU85023

To a solution of 5-chioro-1-et yl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4- di ydroquinoiine-3-carboxylic acid (Example 16f) (5 g, 8.88 mmoi) In N,N- Dimethyliormamide (DMF) (75 mL) was added HATU (3.96 g, 10.42 mmoi) and DIPEA (4.55 mL, 26, 1 mmoi), the resulting mixture was stirred at r.t. for 15 min. Then 2- (pyrroiidln-l-yl)ethanamine (1 ,502 mL, 9,55 mmoi) was added and the resulting mixture was stirred at r.t. for 1 h. The mixture was concentrated and partitioned between water and DCM. The organic layer was separated and washed with water (2 x), sat. NaHCC^ (aq.) and brine. The organic solution was dried over N 2SC>4, and concentrated. The residue was purified by automated silica gel chromatography (CombiFlash Rf) eiuting with MeOH/DCM (0-25%), followed by reverse phase chromatography (CombiFlash Rf) eiuting with ACN/water (0-100%) to afford 5-chioro-1 -ethyi-6,7-bis((4~

met oxybenzyl)oxy)-4-oxo-N-(2-(pyrrol!din-1 -yl)ethyi)-1 ,4-dihydroquinoline-3- carboxamide (3.6 g, 5.81 mmo!, 86.9 % yield) as a white solid. LC S: (M+H)⁺: 620,5

Exampte 27b (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamldo)-3-((1 -(2-(5-chloro-1-ethyl-6,7-dihydiOxy-4-oxo-1 ,4- dihydroqulnoline-3-carboxamido)ethy!)pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5 hla-1 - azabicyclo[4.2.0]oct-2-ene-2-carboxylate, sodium salt

The compound was prepared according to the procedure described In Examples 18k-18! and 17, utilizing 5-chloro-1 -ethyl-6_l7-bis((4-methoxybenzy!)oxy)-4-oxo-N-(2-(pyrro!idin-1- yl)ethyl)-1 ,4-dihydroquinoline-3-carboxamide in place of 5-chloro-1-ethyl-6,7-bis((4- rnethoxybenzyl)oxy)-3-(pyrrolidin-1-ylmethyl)quinolin-4(1 H)-one in Example 16k. LCMS: (M+Hf : 877.6. Ή NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.27 (t, J-7.07 Hz, 3 H) PU85023

2.1 1 (br. s„ 4 H) 2.85 (br. s., 2 H) 3.37 - 3.72 (m, 8 H) 3.76 - 3.89 (m, 2 H) 3.92 - 4.15 (m, 4 H) 4.80 - 4.88 (m, 1 H) 5.20 -5.29 (m, 1 H) 5.74 (d, J=4.55 Hz, 1 H) 6.67 - 6.75 (m, 1 H) 6.80 (s, 1 H) 8.20 - 8.29 (m, 1 H) Example 28 (6R_!7R)-7-((Z)-2-(2-amino-5-criiorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetamido)-3-((1-((1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3- y!)methy!)pyrrGlidin-1-ium-1-y!)methyi)^

carboxyiate, Sodium salt

The compound was prepared according to the procedure from Examples 22f-22g, Liiilizing 1 -et yl-67-bis((4-met oxybenzyi)oxy)-3-(pyrroiidin-1-yl methyl )cinnolin-4(1 H)-one (Example 8f) in place of 5-chloro-1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrro!idin-1- ylmethyl)quinolin-4(1 H)-one in Example 22f. LC S: (M+H)⁺:791.9. H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.28 - 1.47 (m, 9 H) 2.04 (br. s., 4 H) 3.33 - 3.63 (m, 6 H) 3.81 - 3.92 (m, 1 H) 4.10 - 4.22 (m, 1 H) 4.34 (br. s„ 2 H) 4.47 (br. s., 2 H) 5.27 (d, J=5.05 Hz, 1 H) 5.73 - 5.80 (m, 1 H) 6.83 (s, 1 H) 7.17 (s, 1 H) Example 29 (6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino) acetamido)-3-((1-((1 -ethy!-6 J-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-y!) methyl)pyrro!idin-1 -ium-1 -yl)methyi)-8-oxo-5^^

carboxyiate, Sodium sail

The compound was prepared according to the procedure from Examples 211-21 m, utilizing 1-ethyi-6_>7-bis((4~methoxybenzy!)oxy)-3-(pyrrolidin~1 -yimethyl)cinno!in~4(1 H)~one (Example 8f) in place of 5-chloro-1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrro!idin-1- ylmethyi)quinolin-4(1 H)-one in Example 211. LGMS: (M+H)⁺:821 .2. H NMR (400 MHz, PU85023

DEUTERIUM OXIDE) δ ppm 1.37 (t, J=7.07 Hz, 3 H) 2.05 (br. s., 4 H) 2.73 - 2.84 (m, 2H) 3.49 (d, J=16.42 Hz, 6 H) 3.85 (d, J=16.93 Hz, 1 H) 4.18 (s, 1 H) 4.38 (d, J=7.Q7 Hz, 2 H) 4.48 (br. s., 2 H) 4.84 - 4.90 (m, 1 H) 5.25 (d, J=4.80 Hz, 1 H) 5.70 (d, J=4.80 Hz, 1 H) 6,94 (s, 1 H) 7.21 (s, 1 H) )

3oct-2-

The compound was prepared according to the procedure described In Examples 16k-16! and 17, utilizing 1-Ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1 -yl)ethyl)- 1 ,4-di ydrocinnoline-3-carboxamide (Example 1 1 a) in place of 5-chloro-1 -ethyl-6,7-bis((4- methoxybenzyl)oxy)-3~(pyrroiidin-1-ylmeihyi)quinolin-4(1 H)~one in Example 16k. LCMS: (M+H)⁺: 844.1. ^'H N R (400 MHz, DEUTERIUM OXIDE) δ ppm 1.35 (t, J=7.20 Hz, 3 H) 2.12 (d, J=3.79 Hz, 4 H) 2.58 - 2.65 (m, 2 H) 3.36 - 3.64 (m, 8 H) 3.69 - 3.96 (m, 3 H) 4.03 - 4.13 (m, 1 H) 4.32 - 4.45 (m, 2 H) 4.79 - 4.86 (m, 1 H) 5.24 (d, J=5.05 Hz, 1 H) 5.71 (s, 1 H) 6.82 (s, 1 H) 6.94 (s, 1 H) 7.19 (s, 1 H) E am le 31 (6R.7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-3-((1 -(2-(5-chloro-1-ethyl-67-dihydroxy-4-oxo-1 ,4-dihydrocinnolin carboxamido)ethyl)pyrroiidin-1-ium-2-ylium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo

[4.2.0]oct-2-ene-2-carboxylate, Sodium salt

'le_^gl a 1-Ethyl-5-nitro-4-oxo-1 ,4-dihydro-[1 ,3]dioxolo[4,5-g]cinnoline-3-carboxylic acid

To a solution of 1-ethyl-4-oxo-1 ,4-dihydro-[1 ,3]dioxolo[4,5-g]cinnoline-3-carboxylic acid

(5g, 19.07 mmol) in sulfuric acid (15 mL} cooled to 0 °G was added potassium nitrate (2.121 g, 20.97 mmol) in small portions to maintain temperature bellow 10°C with ice water. The mixture was stirred at 10"C for 1 h and at room temperature for 12h, and was then poured into ice cold water (200ml). The mixture was filtered, the solid was washed with water and ethanol to afford 1 -ethy!-5-nitro-4-oxo-1 ,4-d!hydro-[1 ,3jdloxo!o[4,5- g]cinnoline-3-carboxylic acid (4.4g, 14.32 mmol, 75 % yield). LC S: (ivl+H) ^'. 308.2

Example 31 b 5-Amino-1-ethyl-4-oxo-1 ,4-dihydro-[1 ,3]dioxolo[4,5-g]cinnoline-3~carboxyiic acid

To a suspension of 1-ethyl-5-nitro-4-oxo-1 ,4-dihydro-[1 ,3]dioxolo[4,5-g]cinnoline-3- carboxyilc acid (4.25 g, 13.83 mmol) in Acetic Acid (100 mL) and hydrochloric acid (30 mL) in parr shaker bottle was added Pd/C (0.8 g, 5.64 mmol) and the mixture was hydrogenated at 50 psi on a parr shaker for 5h. The mixture was filtered. The filtrate was allowed to drop Into 500 ml of water and the precipitate was collected by filtration, dried to afford 5-amino-1-ethyi-4-oxo-1 ,4-dihydro-[1 ,3]dioxoio[4,5-g]clnnoiine-3-carboxyilc acid (3.8g, 12.99 mmol, 94 % yield). LCMS: (M+Hf: 278.2

Example 31 c 3-Carboxy-1-ethy!-4-oxo-1 ,4-dihydro-[1 ,3]dioxo!o[4,5-g]cinnoiine-5- diazonium chloride

A mixture of 5-amino-1 ~eihyl~4-oxo-1 ,4-dihydro-[1 _!3]dioxoiof4,5-g]cinno!ine-3-carboxylic PU85023 acid (3.6 g, 12.99 mmoi) in hydroch!oric acid (cone) (20 ml) was stirred at room temperature and sodium nitrite (0.986 g, 14.28 mmo!) in water (7 mL) was added dropwise to maintain the temperature beliow 45^!,C. Stirring continued for 5 h. The solution was poured into 50 mi of water and was allowed to stand overnight in the refrigerator. Precipitate was collected by filtration and dried over drierite in a desiccator to yield 3- Carboxy-1-ethyl~4-oxo~1 ,4~dlhydro~[1 ,3]dioxolo[4_>5-g]cinnoline~5-diazonium chloride (3.15 g, 9.67 mmoi, 74.5 % yield). LCMS: (M+H)^÷: 326.2

-Ch loro- 1 -ethy I-6 , 7-d ihyd roxy-4-oxo- 1 ,4-d i hyd roci n nol i ne-3-carboxy I i c acid

A solution of 3-Carboxy-1-ethyl-4-oxo-1 ,4-dihydro-[1 ,3]dloxoio[4,5~g]cinno!ine~5- diazonium chloride (1.85g, 5.68 mmoi) In sulfuric acid (19.98 ml, 187 mmoi) was heated at 95°C for 20 h. Water was added, the precipitates were collected to provide 5-chloro-1 - ethyl-BJ-dihydroxy^-oxo-l ^-dihydrocinno!ine-S-carboxyiic acid (1.46g, 5.13 mmoi, 90 % yield). LCMS: (M+Hf: 285.2

Exarnpfe 31 e 4- ethoxybenzyl 5-chioro-1-ethyi-6.7-bis((4-methoxybenzyl)oxy)-4-oxo- 1 ,4-dihydrocinnoline-3-carboxyiate

To a solution of 5-chlQro-1~ethyl-6,7~rJihydroxy~4-oxo~1 ,4~rJihydrocinnoline-3-carboxyiic acid (1 .46g, 5.13 mmoi) In Ν,Ν-dimethyiformamide (DMF) (20 mL) was added potassium carbonate (1.772 g, 12.82 mmoi) followed by 1-(chioromethyi)-4-methoxybenzene (1.392 mL, 10.26 mmoi). The mixture was heated at 50°C for 12h, extracted with ethyl acetate and washed with water, brine, dried over magnesium sulfate. The solvent was removed under vacuum and the residue was chromatographed on ISCO silica column eiuting with 0-10% methanokdichloromethane to afford 4-methoxybenzyi 5-chloro-1 -ethyl-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoiine-3-carboxylate (1.4g, 2.170 mmoi, 42.3 % yield). LCMS: (M+H)+: 645.2 Exampie 31 f 5-C loro-1 -ethyi-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydrocinnoline- 3-carboxy!ic acid

To the solution of 4-meihoxyhenzy! 5-chioro-1~ethyi-6,7-bis((4-methoxybenzyi)oxy)~4-oxo- 1 ,4-dihydrocinnoline-3-carboxylate (24 g, 37.2 mmol) in methanol (400 mL) and water (133 mL) was added potassium hydroxide (8.35 g, 149 mmol). The reaction mixture was heated to reflux for 3h. Solvent was removed under vacuum. The residue was suspended In water and the resulting mixture was acidified to ρΗ4 with 2N hydrochloric acid. The precipitated solid was filtered and dried to afford 5-chloro-1-ethyl-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoiine-3-carboxylic acid (12.6g, 24.00 mmol, 64.5 % yield). LC S: (M+H)+: 525.2

Example 31 q 5-Chloro-1 -ethyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-N-(2-(pyrroiidin-1-yl) ethyl)- 1 ,4~dlhydrocinnoline-3-carboxamide

To a suspension of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4- dihydroeinnoline-3-carboxyiic acid (3 g, 5.71 mmol) in tetrahydrofuran (THF) (25 mL) was added triethylamine (0.878 mL, 6.29 mmol), followed by isobutyi ch!oroformate (0.826 mL, 6.29 mmol). The reaction mixture was stirred at 0^"C for 30 min. 2-(Pyrroiidin-1- yi)ethanamine (0.979 g, 8.57 mmol) was added and the mixture was stirred at 0"C for 30 min and warmed up to room temperature for 2 h. Solvent was removed and the residue was chromatographed to afford 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N- (2-(pyrrolidin-1 -yl)ethyl)-1 ,4-dihydrocinnoline-3-carboxamide (1.2g, 1.932 mmol, 33.8 % yield). LCMS: (M+H)+: 621 .2

Example 31 h 1-(((6R,7R)-7-((Z)-2-(((1-(tert-butoxy)-2-methyi-1 -oxopropan-2-yl)oxy) imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)acetamido)-2-(((4-m

oxy)carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]od-2-en-3-yl)methyi)-1-(2-(5-ch!oro-1-ethy!- PU85023

6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido)et an-1 -ylium- 1-yl)pyrrolidin-1 -ium

A solution of (5R,6R,7R)-4-methoxybenzyi 7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1 - oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)acetamido)-3- (iodomethyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide (Example 19a) (0.858 g, 0.966 mmoi) and 5-ch!oro-1 -ethy!-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N- (2-(pyrroiidin-1 -yl)ethyl)-1 ,4-di ydrocinnoline-3-carboxamide (0.8 g, 0.966 mmoi) In N,N- dimethylacetamide (DMA) (10 mL) was heated at 40^!,C for 3h, and was then left overnight In the refrigerator. Ν,Ν-dimethylformamide (DMF) ( 0 mL) was added and the mixture was cooied to -40 °C, followed by addition of PBr₃ (0.182 mL, 1.932 mmoi). The mixture was stirred at ~40"C for 30 min, and the reaction was then quenched with 5% NaCI solution. The solid was collected by filtration and purified on ISCO silica gel column eiuting with 0-20% methanokdichioromethane to afford 1 -(((8R,7R)-7-((Z)~2-(((1-(tert- butoxy)-2-methyi-1-oxopropan-2-y!)oxy)lmino)-2-(2-((teri-butoxycarbonyi)amino)thiazoi-4- yi)acetamido)-2-(((4-methoxybenzyi)oxy)carbonyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- en-3-yl)methyi)-1-(2-(5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydrocinnoline-3-carboxamido)ethan-1 -yiium-1-yl)pyrrolidin-1-ium (1.08g, 0.791 mmoi, 82 % yield). LCMS: (M+H)+: 1364.2.

Example 31 i (6R,7R)~7-((Z)-2-(2-amlnothiazol~4-y!)~2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-(2-(5-chloro-1-ethyl-6 J-dihydroxy^-oxo-l ^-dihydrocinnoiine-S- carboxamido)ethyi)pyrroiidin-1-ium-2-yiium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo

[4.2.0]oct-2-ene-2-carboxylate, Sodium salt PU85023

To a solution of 1 -(((6R,7R)-7-((Z)-2-(((1-(tert-butoxy)-2-methyl-1-oxopropan-2- yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-2-(((4- methoxybenzyl)oxy)carbonyl)-8-oxo-5-thia-1-azabicyclo[4.2 ]oct-2-en-3-yi)methyl)-1 -(2- (5-chloro-1-ethyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydrocinnoline-3- carboxamido)et yl)pyrroiidin-1-ium-2-ylium (1.08g, 0,791 mmoi) In dichloromethane (DCM) (20 mL) at room temperature was added anisole (2.59 mL, 23.74 mmoi) foliowed by trifluoroacetic acid (3.05 mL, 39.6 mmoi). The resulting mixture stirred at room temperature for 18 hours, and was then triturated with isopropyi ether. Resulting solid was redissolved in acetonltrile, water and 1 N hydrochloric acid, and HP20ss resin was added. The mixture was concentrated and loaded onto the cartridge containing HP20ss resin. The cartridge was connected to C18 reverse phase ISGO column and eiuted with

0- 95% acetonitrile :water. The fractions with pure desired product were concentrated and lyophilized to constant weight, and was then converted to sodium salt by addition of 1 eq of 0.2N sodium hydroxide, followed by lyophllizatlon to afford (6R,7R)-7-((Z)-2-(2- aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino)acetamido)-3-((1 -(2-(5-chloro-1- ethy i~8 ,7~d i h yd roxy-4~oxG- 1 ,4-d i hyd roe^

ylium-1~yl)methyi)-8-oxo-5~thia-1~azabicycio[4.2.0]oct-2-ene-2-carboxyiate, Sodium salt (150 mg, 0.172 mmoi, 21.78 % yield). LCMS: (M+H)+: 847.3; ¹H NMR (400 MHz,

DEUTERIUM OXIDE) δ ppm 1.31 - 1 ,45 (m, 9 H) 2.13 (br. s., 4 H) 3.58 (br. s„ 6 H) 3.75 - 4, 16 (m, 4 H) 4.49 (d, J=7.58 Hz, 4H) 5.35 (d, J=5.05 Hz, 1 H) 5,67 (s, 1 H), 8.72 (s, 1 H) 6.93 (s, 1 H) Example 32 1 -(((6R,7R)-7-((Z)-2-(2-amino-5-ch!orothiazol-4-yi)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyl)-

1- (2-(5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido)ethan-1- ylium-1-yl)pyrrolidin-1-ium. Sodium salt

The compound was prepared according to the procedure described in Examples 31 h-31 i, utilizing (5R,6R,7R)-benzhydryi 7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1 -oxopropan-2- yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)-5-chlorothiazoi-4-yl)acetamido)-3- (iodomethyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide (Example 22e) in place of (5R,6R,7R)-4-methoxybenzyl 7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1- oxopropan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yl)acetamido)-3- (iodomethyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxlde in Example 31 h. LC S: (M+H)⁺: 882.6. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.31 - 1 .45 (m, 9 H) 2.13 (br. s., 4 H) 3.58 (br. s., 6 H) 3.75 - 4.18 (m, 4 H) 4.49 (d, J=7.58 Hz, 4H) 5.35 (d, 5.05 Hz, 1 H) 5.67 (s, 1 H) 6,93 (s, 1 H)

Exampte 33 (6R,7R)-7-((Z)-2-(2-amino-5-chiorotriiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) lmino)acetamido)-3-((1 -(2~(5-ch!oro-1-ethy!~6,7~dlhydroxy-4~oxo~1 ,4~dlhydroclnnoilne-3- carboxamido)ethyl)pyrrolidin-1-ium-2-ylium-^ ^

[4.2.0]oct-2-en e-2-carboxy late

The compound was prepared according to the procedure described In Examples 31 h-31 i, utilizing (S)-4-tert-butyi 1-(4~methoxybenzyi) 2-(((Z)-(2-(((5R,6R,7R)-2- ((benzhydi loxy)carbonyl)-3-(iodomethyi)-5-oxido-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- en-7-yl)amino)-1 -(2-((tert-butoxycarbonyl)amino)-5-chlorothiazol-4-yl)-2- oxoethylidene)arnino)oxy)suGcinate (Example 21 k) in place of (5R,6R,7R)-4- methoxybenzyl 7-((Z)-2-(((1 -(tert-butoxy)-2-methyl-1-oxopropan-2-yi)oxy)irr!ino)-2-(2- ((tert-butoxycarbonyl)amino)thiazol-4-yi)acetamido)-3-(iodomethy!)-8-oxo-5-thia-1^ azabicyclo[4.2.0]oct-2-ene-2-carboxylate 5-oxide in step 31 h. LCMS: (M+H : 91 1.2. 1 H PU85023

NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .31 - 1.45 (m, 9 H) 2.13 (br. s., 4 H) 3.58 (br. s.,6 H) 3.75 - 4.16 (m, 4 H) 4.49 (d, J=7.58 Hz, 4 H) 5.28 (d, J=5.05 Hz, 1 H) 5.82 (s, 1 H) 7.4 (s, 1 H) Example 34 (6R_!7R)-7-((Z)-2-(2-aminothiazo!-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetam!do)-3-((1 -((5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, Sodium salt

To a yellow suspension of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4- dihydrocinnoline-3-carboxylic add (Example 31f) (6.7 g, 12,25 rnmol) In Tetrahydrofuran (THF) (250 mL) was added TEA (3.42 mL, 24.51 rnmol), followed by careful addition of Isobuty! chioroformate (2.57 mL, 19.60 mmoi). The resulting mixture was stirred at r.t. for 3 h. The crude carbonate solution was then slowly added to NaBH₄ (3.01 g, 80 mmoi) in Ethanoi (150 mL) in an ice bath over 15 min and the resulting mixture was stirred at r.t. for 1 h. The reaction was quenched by addition of (10 mL) and evaporated in vacuo.

The residue was then diluted with water (200 mL) and extracted with dichioromethane (200 mL) twice. The organic extracts were washed with brine and then dried over sodium sulfate, filtered and the resulting filtrate was concentrated in vacuo to afford 5-chloro-1 - ethyi-3-(hydroxymethyi)-6,7-bis((4-methoxybenzy!)oxy)cinnolin-4(1 H)-one (6.5 g, 9.16 mmoi, 74.8 % yield) as a yellow solid, which was used In the next oxidation step without further purification. LC S: (M+H)⁺: 5 .4.

Exam le 34b 5-Chloro-1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydrocinnoline-3-carbaldehyde

To a yellow solution of 5-chloro-1-ethyi-3-(hydroxymethyl)-6,7-bis((4- met oxybenzyl)oxy)cinnoiin-4(1 H)-one (6 g, 8.45 mmol) in Dichloromethane (DCM) (100 mL) was added manganese dioxide (12.1 1 g, 1 18 mmol), and the mixture was stirred at r.t. overnight. Solid was filtered off and washed with DCM. The filtrate was concentrated to afford 5-chioro-1-ethyi-8,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydrocinnoline-3- carba!dehyde (4 g, 4.79 mmol, 56.7 % yield) as a crude dark brown solid. LCMS: (M÷H)^÷: 509.3. Exam pie 34c 5-Chloro-1 -ethyl-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrroiidin-1- y!methyi)cinnoiiri-4(1 H)-orie

To a solution of 5-chioro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydrocinnoline-3-carbaldehyde (3.5 g, 4.28 mmol) in Dichloromethane (DCM) (80 mL) was added pyrrolidine (0.370 mL, 4.48 mmol) and sodium triacetoxyborohydride (1 .355 g, 6.40 mmol). The mixture was stirred at r.t. for 1 h. Then reaction mixture was washed with sat. NaHCC>3 (aq.) and brine, dried over sodium sulfate and the solvent was removed in vacuo. The crude product was then purified through normal phase chromatography (CombiF!ash Rf), eluting with MeOH/DCM (0-20%) to afford 5-chloro-1 -ethyl-8,7-bis((4- methoxybenzyl)oxy)-3-(pyrro!idin-1-yimethyi)cirinoliri-4(1 H)-one (1 .9 g, 3.37 mmol, 79 % yield) as an orange solid. The desired product was dissolved in DCM and washed with sat. aHC03 (aq.) and brine, concentrated and passed through a flash column chromatography before being used for the next step. LCMS: (M+H)⁺: 564.4. Exaropte 34d (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetam!do)-3-((1 -((5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oGt-2-ene-2- carboxyiate, Sodium salt PU85023

The compound was prepared according to the procedure described in Examples 16k-16! and 17, utilizing 5-chioro-1 -ethyi-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrroiidin-1- yimethyl)cinnolin-4(1 H)-one (Example 34c) in place of 5-chloro-1-ethyl-6,7-bis((4- methoxyberizyl)oxy)-3-(pyrrolidiri-1 -yimethyl)quino!iri-4(1 H)-one in step 16k. LCMS:

(M+H)⁺: 821.0. ^'H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.33 (s, 3 H) 2.10 (br. s., 4 H) 2.74 (br. s., 2 H) 3.37 - 3.70 (m, 6 H) 3.80 - 3.94 (m, 1 H) 4.09 - 4.48 (m, 5 H) 4.84 - 4.92 (m, 1 H) 5.24 - 5.33 (m, 1 H) 5.72 (d, J-4.80 Hz, 1 H) 6.83 - 6.74 (m, 1 H) 6.83 - 6.95 (m, 1 H)

?)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)-1 ,2-diearboxyethoxy) lmmo)acetamido)-3-((1 -((5-chloro-1-ethy1-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinno!in-3- yl)methyl)pyrroHdin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- earboxyiate, Sodium salt

The compound was prepared according to the procedure described In Examples 211-21 m, utilizing 5-chioro-1 -ethyi-6 J-bis((4-methoxybenzy!)oxy)-3-(pyrro!idin-1-ylmethyl)cinnoNn- 4(1 H)-one (Example 34c) in place of 5-chloro-1-ethyl-6_>7-bis((4-methoxybenzyl)oxy)-3- (pyrrolidln-1 ~y!methyl)quinolin-4(1 H)-one in step 211. LCMS: (M+H)⁺: 855.0. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.36 (s, 3 H) 2.10 (br. s., 4 H) 2.74 - 2.85 (m, 2 H)

3.49 (br. s., 6 H) 3.81 - 3.92 (m, 1 H) 4.13 - 4.24 (m, 1 H) 4.25 - 4.36 (m, 2 H) 4.44 (br. s., 2 H) 4.85 - 4.92 (m, 1 H) 5.23 - 5.32 (m, 1 H) 5.73 (d, J-4.80 Hz, 1 H) 6.79 (s, 1 H)

Example 36 (6R,7R)-7-((Z)-2-(2-amino-5-chloro hiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1-((1-ethyl-5-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolln~3- yl)methyl)pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5-tnia-1-azabicyclo[4.2.0]oct-2-en carboxyiate, Sodium salt

Example 38a 1-Fiuoro-2,3-dimet oxy-5-nitrobenzene and 2-fluoro-3,4-dimethoxy-1- nitrobenzene

To a solution of nitric acid (853 ml, 1 .91 E+Q4 mmoi) stirred at 0 °C was added dropwlse 1-f!uoro-2,3-dimethoxybenzene (149 g, 954 rnmol). The mixture was stirred at the same temperature for 15 min, and then was allowed to warm up to RT for 15 min. The orange solution was poured into Ice and the resultant solid was filtered, washed with water and dried. LCMS showed that the paie yellow solid was the mixture of two products (ratio 1/1.8). The crude material was purified by reverse phase automatic C18 column chromatography (Comh!fiash RF, 120g column), e!uting with Acetonitrile/Water (0-90%) over 35 min to afford 2-f!uoro-3,4-dlmethoxy-1 -nitrobenzene (53 g, 263 mmoi, 28 % yield) and 1-fluoro-2,3-dimethoxy-5-nitrobenzene (94 g, 467 mmoi, 49 % yield) as white solids. LCMS: (M+Hf: 202.1.

Example 36b 3-Fluoro-4,5~dimethoxyanlline

A mixture of 1 -fiuoro-2,3-dimethoxy-5-nitrobenzene (91 g, 452 mmo!) and piatinum(iV) oxide (9 g, 39.6 mmoi) in ethanol (500 mL) was hydrogenated with a Par shaker (40 psi) at r.t. for 30 min. LCMS indicated completion of the reaction. The mixture was filtered and the filtrate was concentrated to afford 3-fluoro-4,5-dimethoxyaniiine (73 g, 426 mmoi, 94 % yield) as a brown oil that was directly used in next step. LCMS: (M+H)⁺: 171.9.

Exarnpfe 38c Diethyl 2-(((3-fluoro-4,5-dimethoxyphenyl)amino)methyiene)malonate PU85023

To the suspension of 3-fluoro-4,5-dimethoxyaniiine (73 g, 428 mmol) in Ethanol (100 mL) was added diethyl 2-(ethoxymethylene)maionate (85 mL, 428 mmol), and the resluiting mixture was heated at 80 "C for 1 h. LC S indicated completion of the reaction. The mixture was concentrated in vacuo to remove eihanoi, and the residue was washed with hexane to afford diethyl 2-(((3-fluoro-4,5-dimethoxyphenyi)amino)methyler!e)ma!onate (142 g, 418 mmoi, 98 % yield) as a yellow solid. LCMS: ( +H)⁺: 342.1.

Example 38d Ethyi 5-fluoi -6,7-dimethoxy~4-oxo~1 ,4~dihydroquino!ine~3-carboxylate (6a) and ethyl 7-fluoro-5,6-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylate (6b)

F' O

^Μβ0 _{ϊτ Γ}-00₂Η H

To Dowtherm (150 mL) heated at 250 °C was added diethyl 2-(((3-fluoro-4,5- dimet oxyp eny!)amino)met ylene)malonate (30 g, 87 mmol). The mixture was stirred at 250 °C for 30 min. LCMS indicated completion of the reaction. The reaction mixture was allowed to cool a bit and was then added to cold hexane, the precipitates were collected by filtration, washed with hexane and then allowed to dry in the air. Similar scale reactions were repeated several times. From a total amount of the maionate (148 g, 434 mmol), a mixture of ethyl 5-fiuoro-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3- carboxylate and ethyl 7-fluoro-5,6-dimethoxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxyiate was obtained as brown solid without further purification (103 g, 349 mmoi, 80 % yield). LCMS: (M+H)⁺: 296.1.

Example 38e Ethyl 1 -ethyl-5-fluoro-6, ,4-dihydroquinoline-3-carboxylate

A mixture of ethyl 5-fiuoro-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylate and ethyi 7-fluoro-5,6-dimethoxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxylate (103 g, 349 mmo and potassium carbonate (72.3 g, 523 mmoi) in triethyl phosphate (297 mi, 744 mmol) was stirred at 120 °C for 8h. LCMS indicated completion of the reaction. The reaction PU85023 mixture was cooied down to rt and diluted with water, extracted with DCM, dried over sodium sulfate and concentrated in vacuo. The crude materia! was recrysta!llzed in EiOH, the precipitate was collected by filtration and washed by cold EtOH and dried in air.

Another recrystal!ization afforded pure ethyl 1-ethyi-5-fluoro-6,7-dimethoxy-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate (65 g, 201 mmol, 57.6 % yield) as a brown solid. LCMS: (M+Hf: 324.1.

Example 38f 1-Ethyl-5-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquino!ine-3-carboxy!ic acid

To a solution of ethyl 1 -ethyl-5-fluoro-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3- carboxyiate (65 g, 201 mmol) in Dich!oromethane (DCM) (300 mL) was added BBr₃ (95 ml, 1005 mmoi) at -78 °C. The mixture was allowed to warm up to rt, and stirred at 25 °C overnight. LCMS indicated completion of the reaction. The mixture was diluted with eOH and concentrated to dryness. The same process was repeated several times to afford 1-ethyl-5-fiuoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylic acid (48 g, 180 mmol, 89 % yield) as yellow solid. The crude produci was a mixture of acid and small amount of methyl ester, which was used in next step reaction without further purification. LCMS: (M+Hf: 268.0. Exampte 38q 4-Methoxybenzyl 1-ethy!-5-f!uoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate

To a solution of 1~ethyi-5-fiuoro-6,7-dihydroxy-4~Qxo-1 ,4-dihydroquinoiine-3~carbQxylic acid (48 g, 180 mmol) in Ν,Ν-Dimethyiformamide (DMF) (500 mL) was added K₂C0₃ (124 g, 898 mmoi), followed by 1-(ch!oromethyi)-4-methoxybenzene (98 mL, 719 mmol). The reaction mixture was stirred at 50 °C overnight. LCMS indicated completion of the reaction. Water was added and the mixture was stirred at r.t for 15mins. The yellow precipitates were collected by filtration and washed with water to afford 4-methoxybenzyi 1-ethyl-5-fiuoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoiine-3-carboxyiate (68 g, 108 mmol, 60.3 % yield) as a yellow solid. LCMS: (M+H)⁺: 628.4. PU85023

Example 38 1-Ethyi-5-fiuoro-6 -bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline- 3-carboxy!ic acid

To a suspension of 4-methoxybenzy! 1-ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4- oxo-1 ,4-dihydroquinoline-3-carboxyiate (68 g, 108 mmoi) in a mixture of Methanol (250 mL) and Water (125 mL) was added KOH (12.16 g, 217 mmoi} portlonwise. The

resulting mixture was stirred at 90 °C for 3 h. LCMS Indicated completion of the reaction. The reaction mixture was cooled down to r.t. and concentrated, and was then diluted with water and adjusted pH to 1 using 6 N HGI (aq.). The precipitates were collected by filtration and dried to afford 1-ethyl-5-fluoro-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4- dihydroquirioiine-3-carboxylic acid (50 g, 99 mmoi, 91 % yield) as a brown solid. LCMS: (M+Hf: 508.2.

Example 38i 1-Ethyl-5-fluoro-3-(hydroxymethyl)-6,7-bis((4-methoxybenzy!)oxy)quinolin- 4(1 H)-one

To a suspension of 1-ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carboxylic acid (20 g, 39.4 mmoi) In Tetrahydrofuran (THF) (100 mL) was added TEA (6.04 mL, 43.3 mmoi) and isobutyl chloroformate (5.69 mL, 43.3 mmoi). The resulting mixture was stirred at r.t. for 1 h. Then the mixture was cooled down to -78 ^!,C and a solution of DIBAL-H (52.5 mL, 79 mmoi) in toluene (1 .5 M) was added. The mixture was stirred at the same temperature for 2 h, LCMS indicated completion of the reaction. The reaction was quenched with sat. NH₄CI (aq.), and was warmed up to r.t., concentrated, diluted with sat. NH₄CI and extracted with dichioromethane. The organic mxiture was filtered through Celite to remove gummy precipitate. The combined organic solution was then washed with brine, dried over sodium sulfate, filtered and the filtrate was concentrated in vacuo to afford 1- ethyl-5-fluoro-3-(hydroxymethyl)-6_l7-bis((4-methoxybenzy!)oxy)quinolin-4(1 H)-one (1 1.5 g, 23.30 mmoi, 59.1 % yield) as a yellow solid, which was used in the next oxidation step without further purification. LCMS: (M+H)⁺: 494.2 PU85023

Example 36 j 1-Ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline- 3-carbaldehyde

To a yellow solution of 1 -ethyl-5-fluoro-3-( ydroxymet yl)-6,7-bis((4-methoxybenzyi)oxy) quinolin-4(1 H)-one (1 1 g. 22.29 mmol) in Dichloromethane (DCM) (100 mL) was added manganese dioxide (19.38 g, 223 mmol). The mixture was stirred at rt for 8.5 h. The mixture was then filtered through Gelite and washed with DCM. the filtrate was concentrated to afford 1 -ethyl-5-flL!oro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carbaldehyde (9.8 g, 19.94 mmol, 89 % yield) as a yellow solid, which was used in the next step without further purification. LCMS: (M+H)⁺: 492.2.

Example 38k 1-Ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1-yimethyi) quinolin-4(1 H)-one

To a solution of 1-ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 _>4- dihydroqLiinoiine-3-carbaldehyde (18 g, 36.6 mmol) In 1 ,2-Dichloroethane (DCE) (100 mL) was added pyrrolidine (4.53 mL, 54.9 mmol), sodium trlacetoxyborohydrlde (15.52 g, 73.2 mmol) and AcOH (0.105 mL, 1.831 mmol) . The reaction mixture was stirred at 25 °C for 3h. LCMS indicated completion of the reaction. The mixture was extracted with DCM and washed with brine. The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude material was purified by normal phase automatic silica column chromatography (Combinfiash RF), using a 40g gold column and eiuting with MeOH/DCM (0-20%). The product was further purified by reverse phase automatic chromatography (Combinfiash RF), using a 150g gold C18 column and eiuting with acetonitrile/Water (0%-60%) to afford 1 -ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-3- (pyrrolidin~1 ~yimethyl)quinolin-4(1 H)-one (9.4 g, 17.20 mmol, 47.0 % yield) as a pale yellow solid. LCMS: (M+H)⁺: 547.3. Example 38S (6R,7R)-7-((Z)-2-(2-amino-5-chiorothiazo!-4-yi)-2-(((S)-1 ,2-diearboxyethoxy) imino)acetamido)-3-((1-((1-ethyl-5-fluoro-6,7-dihydroxy-4-oxo-1 .4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyl)-8-oxo-5 hla-1-azabicycio[4.2.0]oct-2-ene-2- earboxylate, Sodium sai

The compound was prepared according to the procedure described in Examples 211-21 m, utilizing 1 -ethyi-5-f iuoro-6 -bis((4-methoxybenzyl)oxy)-3-(pyrroiidin-1-ylrnethyl)quinoiin- 4(1 H)-one (Example 36k) in place of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-3- (pyrrolidin-1 -ylmethyi)quinolin-4(1 H)-one in step 21 i. LCMS: ( +H)⁺: 855.0. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.31 (t, J=7.07 Hz, 3 H) 2.02 (br. s., 4 H) 2.76 (br. s., 2 H) 3.24 - 3.47 (m, 6 H) 3.82 - 3.93 (m, 1 H) 3.94 - 4.04 (m, 1 H) 4.07 - 4.19 (m, 2 H) 4.20 - 4.38 (m, 2 H) 4.83 - 4.91 (m, 1 H) 5.22 - 5.31 (m, 1 H) 5.88 - 5.78 (m, 1 H) 6.75 - 6.83 (m, 1 H) 8.06 (s, 1 H)

Exampte 37 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yi)oxy) imino) acetamido)-3-((1-((1-ethyi-5-fluoro-6 J-dity

Dyrrolidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-erie-2-carboxylate

The compound was prepared according to the procedure described in Examples 19b-19c, utiiizing 1-ethyi-5-fiuoro-6_!7-bis((4-met oxybenzyi)oxy)-3-(pyrroiidin-1-ylmethyl)quinoiin- 4(1 H)-one (Example 36k) in place of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-3- (pyrrolidin-1 -yimethyi)quinolin-4(1 H)-one in step 19b, LCMS: (M+H)⁺: 774.0. ^!H NMR (400 MHz, METHANOL-^) δ ppm 1 .50 (t, J=7.07 Hz, 3 H) 1.61 (d, J=6.32 Hz, 6 H) 2.1 1 - 2.30 (m, 4 H) 3.42 - 3.65 (m, 6 H) 3.92 - 4.02 (m, 1 H) 4.07 - 4.17 (m, 1 H) 4.26 - 4,39 (m, 3 H) 4.46 - 4.55 (m, 1 H) 5.27 - 5.34 (m, 1 H) 5.92 (d, J=5.05 Hz, 1 H) 6.89 (s, 1 H) 6.91 - 6.96 (m, 1 H) 8.18 - 8.25 (m, 1 H).

Example 38 (6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-fluouro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yl) methyl)pyrrolidin-1 -ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate, Sodium salt PU85023

The compound was prepared according to the procedure described In Examples 16k-16! and 17, utilizing 1-ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1- ylmethyi)quinolin-4(1 H)-one (Example 36k) in place of 5-chioro-1 -ethyi-6,7-bis((4- methoxyberizyl)oxy)-3-(pyrrolidin-1-yimethyi)quinolin-4(1 H)-one in step 16k. LCMS:

(M+H)⁺: 804.3; ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .31 - 1 .45 (m, 9 H) 2.13 (br, s., 4 H) 2.7 (br.s 2H), 3.48 (br. s.,6 H) 3.75 - 4,16 (m, 4 H) 4,49 (d, J=7.58 Hz, 4 H) 5.31 (d, 5.05 Hz, 1 H) 5.75 (s, 1 H) 8.80 (s, 1 H) 7.90 (s, 1 H), 8.05 (s, 1 H). ct-2~

Example 39a 1-Ethyl-5-flu -oxo-N-(2-(pyrrolidin-1 - yi)ethyl)-1 ,4-dihydroquino!i

To a solution of 1-ethyl-5-fluoro-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydro quinoline-3-carboxylic acid (Example 38h) (5 g, 9.85 mmoi) in N.N-Dimethylformamide (DMF) (5 mL) was added HATU (5.62 g, 4.78 mmoi) and DIPEA (5.16 mL, 29.6 mmoi), the resulting mixture was stirred at r.t. for 30 min. Then 2-(pyrrolidin-1-yi)ethanamine (1.861 mL, 14.78 mmoi) was added and the resulting mixture was stirred at r.t. overnight. The mixture was concentrated and the residue was purified by normal phase automatic silica column chromatography (Combiflash RF) eluting with MeOH/DCM (0-20%). The product was further purified by reverse phase automatic chromatography (Combinflash RF), using a 150g golden C18 column and eluting with acetonitrile/VVater (0%-100%) to afford 1-ethyl-5-fiuoro-6_!7-bis((4-methoxybenzyi)oxy)-4-oxo-N-(2-(pyrro!idin-1-yl)ethyl)- 1 ,4-dihydroquinoiine-3-carboxamide (3.8 g, 6.29 mmol, 83.9 % yield) as pale vvviiovv solid. LC S: (M+H)⁺: 604.4.

Example 39b (6R_l7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyet oxy) imlno)acetamldo)-3~((1~(2-(1 ~ethyl-5-fluoro-6_>7-dihydroxy-4~oxo-1 _>4-dihydroqLjinoiiRe-3~ carboxamido)ethy!)pyrroiidin-1 -ium-1 -y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate, sodium salt

The compound was prepared according to the procedure described In Examples 18k-18! and 17, utilizing 1-ethyl-5-fluoro-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-N-(2-(pyrro!idin-1- yl)ethyl)-1 ,4-dihydroquinoline-3-carboxamide (Example 39a) in place of 5-chloro-1 -ethyl- 6,7-bis((4-methoxybenzyl)oxy)-3-(pyrroiidin-1 -ylmethyl)quinoiin-4(1 H)-one in step 16k. LCMS: (M+H)⁺: 861.2; ¹H NMR (400 MHz, DEUTERIUM OXIDE) p^" ppm 1.30 (t, J=6.82 Hz, 3 H) 2.1 1 (br. s., 4 H) 2.61 (br. s., 2 H) 3.36 - 3.75 (m, 8 H) 3.83 (d, J=16.93 Hz, 2 H) 4.00 - 4.18 (m, 4 H) 4.83 (s, 1 H) 5.19 - 5.28 (m, 1 H) 5.72 (d, J=4.80 Hz, 1 H) 6.64 - 6.73 (m, 1 H) 6.83 (s, 1 H) 8.35 (s, 1 H)

Exampte 40 (6R,7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imirio)acetarriido)-3-((1-((1-ethyl-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dih droquinolin-3- yi)methyl)pyi oiidin-1-ium-1-yl)methyl)-8-oxo-5 hla-1~azabicyclo[4.2.0]oct-2-ene~2- carboxylate, Sodium salt

PU85023

Exam le 40a 1-Ethyl-6-fluoro-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline- 3-carboxyiic acid

A mixture of ethyl 1 -ethyi-6,7,8-trifluoro-4-oxo-1 ,4-dihydroquinoiine-3-Garboxylate (50 g, 167 mmol), KOH (94 g, 1871 mmol) and 4-methoxybenzy! alcohol (208 ml, 1671 mmol) was heated at 75 ^CiC for 4 hours. LCMS showed completion of reaction. The solution was adjusted to pH 2 and was extracted with ethyl acetate. The organic layer was washed with water, dried with Na₂S0₄, filtered, and concentrated in vacuo. The residue was triturated with ethyl acetate, whereupon the crude product precipitated out as a solid substance. The precipitate was collected by filtration and washed with EA to afford 1- ethyl-6-fiLioro-7,8-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydroquinoi!ne-3-carboxylic acid (44 g, 87 mmol, 51.9 % yield) as a yellow solid. LCMS: (M+Hf: 508.3.

Example 40b 1-Ethyi-6-fiuoro-3-(hydroxymet yi)-7,8-bis((4-methoxybenzyl)oxy)quinoiin- 4(1 H)-one

To a suspension of 1-et yl-6-fluoro-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroqulnoline-3-carboxyilc acid (40 g, 79 mmol) In Tetrahydrofuran (THF) (300 mL) was added TEA (12.08 mL, 87 mmol) and isobutyl chloroformate (1 1.39 mL, 87 mmol). The resulting mixture was stirred at r.t. for 1 h. LCMS showed completion of the reaction. Then the mixture was cooled down to -78 °C and a solution of DIBAL-H (158 mL, 236 mmol) In toluene (1.5 M) was added. The mixture was stirred at the same temperature for 1.5 h, LCMS indicated completion of the reaction. The reaction was quenched with sat. NH4CI (aq.), warmed up to r.t., concentrated, diluted with sat. NH₄CI and extracted with dich!oromethane twice. The organic mxiture was filtered through Celite to remove any solids. The combined organic solution was then washed with brine and dried over sodium PU85023 sulfate, filtered and the resulting filtrate was concentrated in vacuo to afford 1-ethy!-6- fiuoro-3-(hydroxymethyl)-7,8-bis((4-methoxybenzyl)oxy)quinoliri-4(1 H)-one (33 g, 51.5 mmoi, 85.3 % yield) as yel!ow solid, which was used in the next oxidation step without further purification, LC S: (M+H)⁺: 494.3.

Example 40c 1-Ethyl-6-fluoro-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline- 3-carbaldehyde

To a yellow solution of 1 -ethyi-8-fiuoro-3~(hydroxymethyl)~7,8-bis((4-methoxybenzyl)oxy) quinolln-4(1 H)-one (33 g, 88.9 mmoi) in Dichloromethane (DCM) (200 mL) was added manganese dioxide (58.1 g, 669 mmoi), and the mixture was stirred at rt for 6.5 h. The mixture was then filtered through Ceiite and the solid was washed with DCM. The filtrate was concentrated in vacuo to afford 1 -ethyl-6-fluoro-7,8-bis((4-methoxybenzyl)oxy)-4- oxo-1 ,4-dihydroquinoline-3-carbaldehyde (25 g, 50.9 mmoi, 78 % yield) as a yeiiow solid, which was used In the next step without further purification. LCMS: (M+H)⁺: 492.3.

Example 40 d 1-Ethy!-6-f!uoro-7,8-bis((4-methoxybenzy!)oxy)-3-(pyrroiidin-1-ylmethyl) quinolin-4(1 H)-one

To a solution of 1-ethyl-6-fluoro-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroqulnoline-3-carbaldehyde (15 g, 30.5 mmoi) In 1 ,2-Dichioroethane (DCE) (100 mL) was added pyrrolidine (3.78 mL, 45.8 mmoi), sodium triacetoxyborohydride (12.94 g, 61.0 mmoi) and AcOH (0.087 mL, 1.528 mmoi). The reaction mixture was stirred at 25 ^GC for 3h. LCMS showed completion of the reaction. The mixture was extracted with DCM and washed with brine. The organic solution was dried over sodium sulfate and evaporated in vacuo. The crude materia! was purified by normal phase automatic silica PU85023 column chromatography (Combifiash RF), using a 40g golden column and eiuting with MeOH/DCM (0-20%). The product was further purified with reverse phase automatic silica column chromatography (Combifiash RF), using a 150g golden column and eiuting with acetonltriie/water (0-80%) to afford 1-ethyi-6-fluoro~7,8~bls((4~methoxybenzyl)oxy)-3~ (pyrrolidin-1 -yimethyi)quinolin-4(1 H)-one (9,8 g, 17.93 mmol, 58.7 % yield) as a yellow solid. LC S: (M+H)⁺: 547.4.

Example 40e (6R_>7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((S)-1 _>2-dicarboxy ethoxy)imino)acetamido)-3-((1-((1 -ethyi-6-fiuoro-7,8-dihydroxy-4-oxo-1 ,4~dihydroquinolin- 3~yl)methyl)pyr! l!d!n-1-ium-1 -yl)methy!)-8-oxo-5-thia-1 ~azabicyc!o[4.2.0]oct~2-ene~2- carboxyiate, Sodium salt

The compound was prepared according to the procedure described in Examples 21 l-21 m, utilizing 1-ethyi-6-fiuoro-7,8-bis((4-methoxybenzyi)oxy)-3-(pyrroiidin-1-ylmet yl)quinoiir!- 4(1 H)-one (Example 40d) in place of 5-ch!oro-1-ethy!-6,7-bis((4-met oxybenzyl)oxy)-3- (pyrrolidin-1 -ylmethyi)quinolin-4(1 H)-one in step 211. LCMS: (M+H)⁺: 838.8. ^!H NMR (400 MHz, DEUTERIUM OX!DE) δ ppm 1.32 (t, J-6.95 Hz, 3 H) 2.01 (br. s., 4 H) 2.69 - 2.82 (m, 2 H) 3.40 (d, J-18.93 Hz, 8 H) 3.80 - 3.92 (m, 1 H) 3.94 - 4.06 (m, 1 H) 4.23 - 4.44 (m, 2 H) 4.53 - 4.64 (m, 2 H) 4.86 (s, 1 H) 5.22 - 5.29 (m, 1 H) 5.71 (d, J-5.05 Hz, 1 H) 7.43 - 7.50 (m, 1 H) 8.10 (s, 1 H)

Example 41 (6R,7R)-7-((Z)-2~(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-((1-ethyi-6-f!uoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi) pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate

PU85023

The compound was prepared according to the procedure described in Examples 19b-19c, utilizing 1-ethyl-6-fluoro~7,8~bis((4-methoxybenzyl)oxy)-3-(pyrrol!din-1-ylm

4(1 H)-one (Example 40d) In place of 5-ch!oro-1-ethy!-6,7-bls((4-methoxybenzyl)oxy)-3~ (pyrrolidin-1 -ylmethy!)quinolin-4(1 H)-one in step 19b, LC S: ( +H)⁺: 773.9. ^!H N R (400 MHz, METHANOL-d4) δ ppm 1.51 (br. s., 9 H) 2,20 (br. s., 4 H) 3.50 (d, J=1.52 Hz, 6 H) 3.91 - 4.04 (m, 1 H) 4.07 - 4.20 (m, 1 H) 4.31 - 4.44 (m, 1 H) 4.48 - 4.61 (m, 1 H) 4.69 - 4.83 (m, 2 H) 5.27 - 5.35 (m, 1 H) 5.86 - 6.01 (m, 1 H) 6.82 - 6.96 (m, 1 H) 7.55 - 7.69 (m, 1 H) 8.17 - 8.34 (m, 1 H)

Exarnpte 42 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-ethyi-6-fiuoro-7,8-dih^

methyl)pyrrolidin-1 -ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]od-2-ene-2- carboxyiate, sodium sal

The compound was prepared according to the procedure described In Examples 16k-16l and 17, utilizing 1~ethyi-6-fiuoro~7,8-bis((4~methoxybeRzyi)oxy)-3~(pyri ildin-1 - y!methyi)quinolin-4(1 H)-one (Example 40d) in place of 5-chloro-1-ethyl~6,7-bis((4~ methoxybenzyl)oxy)-3-(pyrro!idin-1 -yimethyl)quinoiin-4(1 H)-one in step 16k. LCMS:

(M+H)⁺: 804.1. ^!H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .31 (t, J-6.95 Hz, 3 H) 2.02 (br. s., 4 H) 2.62 - 2.75 (m, 2 H) 3.18 - 3.48 (m, 6 H) 3.80 - 3.91 (m, 1 H) 3.95 - 4.07 (m, 1 H) 4.22 - 4.44 (m, 2 H) 4.52 - 4.63 (m, 2 H) 4.83- 4.90 (m, 1 H) 5.21 - 5.31 (m, 1 H) 5.68 - 5.75 (m, 1 H) 6.90 (s, 1 H) 7.47 (d, J=10.61 Hz, 1 H) 8.07 - 8.18 (m, 1 H}

Example 43 (6R,7R)-7-((Z)-2-(2-aminot iazoi-4-yl)-2-(((2-carboxypropari-2-yi)oxy) imino)acetamido)-3-((1-(2-(1 -et yi-6-fiuoro-7,8-di ydroxy-4-oxo-1 ,4-di ydroquino carboxamido)ethyl)pyrroiidin-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2-carboxyiate

PU85023

Exam le 43a 1-Ethyl-6-fluoro-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1 - yi)ethyl)-1 ,4-dihydroquinoline-3-carboxamide

To a solution of 1-ethyl-6-fluoro-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 .4- dihydroquinoiine-3-carboxylic acid (Example 40a) (8 g, 15.76 mmol) In N,N- Dimethylformamide (DMF) (15 mL) was added HATU (8.99 g, 23.64 mmol) and DIPEA (8.26 mL, 47.3 mmol), the resulting mixture was stirred at r.t. for 30 min. 2-(Pyrrolidin-1- yl)ethanamine (2.98 mL, 23.64 mmol) was added and the resulting mixture was stirred at r.t. for 3 h. The mixture was concentrated and the residue was purified by normal phase automatic silica column chromatography (Combiflash RF) eluting with MeOH/DC (0- 20%) to afford 1-ethyi-6-fiuoro-7_!8-bis((4-methoxybenzyi)oxy)-4-oxo-N-(2-(pyrroiidin-1- yl)ethyl)-1 ,4-dihydroquino!ine-3-carboxamide (6.6 g, 0.93 mmol, 69.4 % yield) as a pale yellow solid. LCMS: (M+H)⁺: 604.5.

Example 43b (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-3-((1-(2-(1 -ethyi-6-fiuoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinolin carboxamldo)eihy!)pyrrolidin-1~ium-1~y!)methyi)-8-oxo-5-thia~1-azabicyclo[4.2.0]oct-2~ ene-2-carboxyiate

The compound was prepared according to the procedure described In Examples 19b-19c, utilizing 1-ethyi-6-fiuoro-7_!8-bis((4-methoxybenzyi)oxy)-4-oxo-N-(2-(pyrroiidin-1 -yl)ethy!)- 1 ,4-dihydroquinoiine-3-carboxamide (Example 43a) In place of 5~ehioro~1 -ethyi-8,7-bis((4~ methoxybenzyl)oxy)-3-(pyrro!idiri-1-yimethyi)quinolin-4(1 H)-one in step 19b, LCMS:

(M+Hf: 832.2. ¹H NMR (400 MHz, METHANOLS) <5 ppm 1 .61 (d, 3.54 Hz, 9 H) 2.15 - 2.32 (m, 4 H) 3.46 - 3.73 (m, 6 H) 3.87 - 4.07 (m, 3 H) 4.1 1 - 4.22 (m, 1 H) 4.76 - 4.85 (m, PU85023

2 H) 5.27 - 5.33 (m, 1 H) 5.88 - 5.96 (m, 1 H) 6.85 - 6.92 (m, 1 H) 7.65 - 7.73 (m, 1 H) 8.64 - 8.71 (m, 1 H)

Example 44 (6R JR)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyetrioxy)irriino) acetamido)-3-((1-((1-ethyi-7,8-dihydroxy-4-oxo-1 ,4-di ydroquinoiin-3-yl)meth

1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-Garboxylate_! sodium salt

Example 44a Diethyl 2-(((2,3-dimethoxyphenyl)amino)methyiene)malonate

To a mixture of 2,3-dimethoxyanilirie (20 g, 131 mmoi) in Ethano! (50 mL) was added diethyi 2-(ethoxymethyiene)maionate (26.2 mL, 131 mmoi). The mixture was heated at 80 °C for 1 h. LCMS indicated compietion of the reaction. The mixture was concentrated in vacuo to remove ethanol, and washed with hexane to afford diethyi 2-(((2,3- dimethoxypheny!)amino)methylene)malonate (38 g, 1 18 mmoi, 90 % yield) as a white solid. This crude mixture was directly used in next step. LCMS: (M+H)⁺: 324.2.

E am le 44b Ethyl 7,8-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxyiate

To Dowtherm A ( 00 mL) heated at 250 °C was added diethyi 2-(((2.3-dimethoxyphenyi) amino)methylene)malonate (48 g, 148 mmoi). The mixture was stirred at 250 °G for 1 h. LCMS Indicated completion of the reaction. The reaction mixture was allowed to cool a bit and was then added to cold hexane, the precipitates were collected by fl!tration, washed with hexane and then allowed to dry in the air to afford ethy! 7,8~dimethoxy-4-oxo-1 ,4- dihydroquirioiine-3-carboxylate (35 g, 126 mmoi, 85 % yield) as brown solid. LCMS: (M+Hf: 278.2. PU85023

Example 44c Ethyl 1 -ethyl-7,8-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylate

A mixture of ethyl 7,8-dimethoxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxylate (40 g, 144 mrnoi) and potassium carbonate (29.9 g, 216 mrnoi) in triethyi phosphate (123 mi, 721 mmo!) was stirred at 120 °C for 24 h. LC S indicated completion of the reaction. The reaction mixture was cooled down to rt and diluted with water, extracted with DCM, dried over sodium sulfate and evaporated in vacuo. The crude material was recrystaliized in EtOH, the precipitate was collected by filtration, washed by cold EtOH and dried in air to afford ethyl 1-ethyl-7,8-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylate (26 g, 85 mmol, 59.0 % yield) as a yellow solid. LCMS: (M+H)⁺: 306.2.

Example 44d Ethyl 1-ethyl-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxyiate and methyl 1-ethyi-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxylate

To a solution of ethyl 1 -ethyl-7,8-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylate (12 g, 39.3 mmo!) in Dichioromethane (DCM) (200 mL) was added BBr₃ (18.58 mL, 197 mmo!) at -78 °C. The mixture was allowed to warm up to rt, and stirred at rt for 5h. LCMS indicated completion of the reaction and products were a mixture of ethyl and methyl esters. The mixture was diluted with MeOH and concentrated. This procedure was repeated for several times to afford a mixture of ethyl 1-ethyi-7,8-dihydroxy-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate and methyl 1-ethyi-7,8-dihydroxy-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate as yellow solid (ratio 1 :1 .5, 10 g, 36.1 mrnoi, 92 % yield). The crude mixture was used in next step without further purification. LCMS: (M÷H)^÷: 278.1 (ethyl ester): 264.1 (methyl ester).

Exampte 44e Methyl 1-ethyl-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoiine- 3-carboxyiate, ethyl 1-ethyi-7_!8-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-dihydroquinoline~3~ carboxylate and 4~methoxybenzyi 1 -ethyi-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carboxylate PU85023

To a solution of the mixture from Example 44d (28 g, 106 mmol) in N,N- Dimethyiformamide (DMF) (200 mL) was added K₂C0₃ (73.5 g, 532 mmol), followed by 1~(chioromethyi)~4-methoxybenzene (43.3 mL, 319 mmol). The reaction mixture was stirred at 50 °C for 4h. LCMS indicated completion of the reaction and that the product was a mixture of Methyl, Ethyl and PMB esters. Water was added and the mixture was stirred at r.t for 15 min. The precipitates were collected by filtration and washed with water to afford a mixture of methyl 1-ethyl-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-S-earboxylate, ethyl 1-ethyi-7,8-bis((4-methoxybenzyi)oxy)~4-oxo-1 ,4- dihydroquinoiine-3-carboxylate and 4-methoxybenzyl 1-ethyi-7,8-bis((4-methoxy benzyi)oxy)-4-oxo-1.4-dihydroquinoline-3-carboxylate (ratio 1 :2 : 1.5, 45 g, 73.8 mmol, 69.4 % yield) as a brown solid. The crude mixture was used in next step without further purification. LCMS: (M+H)⁺: 504.3 (methyl ester); 518.3 (ethyl ester); 6 0.4 (PMB ester). Exampje 44f 1-Ethyl-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroqiiinoline-3- carboxyilc acid

To a suspension of the mixture from Example 44e (45 g, 73.8 mmol) in a mixed solvent of Methanol (250 mL) and Water (125 mL) was added KOH (12.42 g, 221 mmol) portlonwise. The resulting mixture was stirred at 90 ⁼C for 3 h. LCMS indicated completion of the reaction. The reaction mixture was cooled down to r.t. and

concentrated, and was then diluted with water and adjusted pH to 1 using 6 N HC! (aq.). The precipitates were collected by filtration and dried to afford 1-ethyi-7,8-bis((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3-carboxylic acid (35 g, 71 .5 mmol, 97 % yield) as pale yellow solid. LCMS: (M+H)⁺: 490.3.

Example 44g 1-Ethyl-3-(hydroxymethyl)-7,8-bis((4-methoxybenzyl)oxy)quinolin-4(1 H)-one PU85023

To a suspension of 1-ethyl-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxylic acid (22 g, 44,9 mmoi) in Tetrahydrofuran (THF) (300 mL) was added TEA

(6.89 mL, 49.4 mmoi) and isobutyi chioroformate (6.49 mL, 49.4 mmoi). The resulting mixture was stirred at r.t. for 1 h. Then the mixture was cooled down to -78 °C and a solution of DIBAL-H (90 mL, 135 mmoi) in toluene (1 .5 M) was added. The mixture was stirred at the same temperature for 1 .5 h, LCMS indicated completion of the reaction. The reaction was quenched with sat. NH₄CI (aq.), and subsequently warmed up to r.t.,

concentrated, diluted with sat. NH₄CI and extracted with dichloromethane twice. The organic phase was filtered through Celite to remove gummy precipitate, and was then washed with brine, dried over sodium sulfate, filtered and concentrated in vacuo to afford 1-ethy!-3-(hydiOxymethyi)-7,8-bis((4-methoxybenzyl)oxy)quinoi!n-4(1 H)-orie (20 g, 23.55 mmo!, 52.4 % yield) as yellow solid, which was used in the next oxidation step without further purification, LCMS: ( +H)⁺: 476.3.

Example 44 1-Ethy!-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquino!ine-3-carbaldehyde

To a yellow solution of 1-ethyl-3-(hydroxyrnethyl)-7,8-bis((4-methoxybenzyl)oxy)quinolin- 4(1 H)-one (16 g, 33.6 mmo!) in Dichloromethane (DCM) (200 mL) was added

manganese dioxide (29.3 g, 336 mmoi), and the mixture was stirred at rt for 6.5 h. The mixture was filtered through Celite and washed with DCM, the filtrate was concentrated to afford 1~ethyi-7,8-bis((4-methoxybenzyi)oxy)~4-oxo~1 ,4~rJihydroqulnoline-3-carbaidehyde (10 g, 21 .12 mmoi, 62.8 % yield) as a brown solid, which was used in the next step

without further purification. LCMS: (M-+H)^*: 474.3.

Example 44 i 1-Ethyl-7_>8-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1 -ylmethyi)quinolin-4(1 H)-one

PU85023

To a solution of 1-ethyl-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carba!dehyde (10 g, 21.12 mmol) in Dichloromethane (DCM) (150 mL) was added pyrrolidine (2.81 mL, 31.7 mmol), sodium triacetoxyborohydride (8.95 g, 42.2 mmo!) and AcOH (0.060 mL, 1 ,056 mmol). The reaction mixture was stirred at 25 °C for 3 h. LCMS showed completion of the reaction. The mixture was extracted with DCM and washed with brine. The organic layer was dried over sodium sulfate and evaporated in vacuo. The crude material was purified by normal phase automatic silica column chromatography (Combifiash RF), using a 120g column and elutlng with MeOH/DC (0-20%). The product was further purified with reverse phase automatic silica column chromatography (Combifiash RF), using a 150g golden column and eluting with acetonitn!e/water (0-80%) to afford 1 -ethyl-7,8-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1-yimethyl)quinolin-4(1 H)- one (4.6 g, 8.70 mmol, 41.2 % yield) as a pale yellow solid. LCMS: ( +H)⁺: 529.4.

Example 44j (6R_!7R)-7-((Z)-2-(2-aminothiazol-4-y!)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-ethyi-7_!8-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyl)pyrrolidin 1-ium-1-y!)metnyl)-8-oxo- rboxylate, sodium salt

The compound was prepared according to the procedure described In Examples 16k-16! and 17, utilizing 1-ethyl-7_!8-bis((4-methoxybenzyi)oxy)-3-(pyrrolidin-1-ylmethyl)quinolin- 4(1 H)-one (Example 44!) in place of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-3-

(pyrrolidin-1 -y!methyi)quinolin-4(1 H)-one in step 16k. LCMS: (M+H)⁺: 786.4. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.32 (s, 3 H) 1.91 - 2.10 (m, 4 H) 2.59 (s, 2 H) 3.22 - 3.50 (m, 6 H) 3.80 - 3.89 (m, 1 H) 3.97 - 4.06 (m, 1 H) 4.23 - 4.34 (m, 1 H) 4.35 - 4.44 (m, 1 H) 4.56 - 4.65 (m, 2 H) 4.84 (dd, J=8.21 , 4.93 Hz, 1 H) 5.22 - 5.32 (m, 1 H) 5.69 - 5.77 (m, 1 H) 6.89 (s, 1 H) 7.09 (d, J=9.09 Hz, 1 H) 7.66 - 7.75 (m, 1 H) 8.08 - 8.16 (m, 1 H)

E am le 45 (6R.7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-3-((1-(2-(1 -ethyl-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2- ene-2-carboxylate PU85023

Example 45a 1-Ethyl-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1 -yl)ethyi)- 1 , 4-d i hyd roq u i no! i n e-3-ca rboxa m i d e

To a solution of 1~ethyi-7,8-bis((4-methoxybenzyi)oxy)~4-oxo-1 ,4-dihydroquinollne-3- carboxyiic acid (Example 44f) (7 g, 14.30 mmoi) in N,N-Dlmethyiformamide (DMF) (50 mL) was added HATU (8.16 g, 21 .45 mmoi) and DIPEA (7.49 mL, 42.9 mmoi), the resulting mixture was stirred at r.t. for 30 mln. 2-(Pyrroiidin-1-yl)ethanamine (2.70 mL, 21.45 mmoi) was added and the resulting mixture was stirred at r.t. for 3h. The mixture was concentrated and the residue was purified by normal phase automatic silica column chromatography (Combifiash RF), eluting with MeOH/DCM (0-30%) to afford 1-ethyl-7,8-bis((4- methoxybenzyl)oxy)-4-oxO-N-(2-(pyrroildin~1 -yl)ethyl)-1 ,4-dihydroquinoline-3-carboxamide (6.6 g, 1 1.27 mmoi, 79 % yield) as a pale yellow solid. LC S: (M+H)⁺: 586.5.

Example 45b (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetamido)-3-((1-(2-(1-ethyl-7^-dihydiOxy-4-oxo-1 _!4-dihydroquinolirie-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate

The compound was prepared according to the procedure described In Examples 19b-19c, utilizing 1-Ethyi-7,8-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1-yi)ethyl)-1 ,4- dihydroquinoiine-3-carboxamide (Example 45a) in place of 5-ch!oro-1 -ethy!-6,7-bis((4- methoxyberizyl)oxy)-3-(pyrrolidiri-1-yimethyi)quinolin-4(1 H)-one in step 19b. LC S: PU85023

(M+H)⁺: 813.4. ¹H N R (400 MHz, METHANOLS) δ ppm 1.52 (t, J=6.95 Hz, 3 H) 1.57 - 1.66 (m, 6 H) 2.13 - 2.32 (m, 4 H) 3.47 - 3.74 (m, 8 H) 3.84 - 4.21 (m, 4 H) 4.78 - 4.87 (m, 2 H) 5.27 - 5.33 (m, 1 H) 5.88 - 5.96 (m, 1 H) 6.86 - 6.93 (m, 1 H) 7.05 - 7.13 (m, 1 H) 7,86 - 7.93 (m, 1 H) 8.63 - 8.70 (m, 1 H)

Example 48 (6R_!7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropari-2- y!)oxy)imino)aceiamido)-3-((1-(2~(1-ethyi~7,8-dihydroxy-4~oxo-1 ,4-di

carboxamido)ethy!)pyrroiidin-1-ium-1-y!)^

ene-2-carboxylate, Sodium salt

(6RJR)-7-((Z)-2-(2-aminothiazoW-yl)-2-(((2-carboxypropan-2-yl)oxy)imino)acetamido)-3- ((1-(2-(1-ethyl-7.8-dihydroxy-4-oxo-1.4-dihydroquinoline-3-carboxamido)ethyi)pyrrolidi iLim-1 -yl)methyi)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2-carboxyiate (Example 45b) (0.163 g, 91 % pure, 0.182 mmo!) was suspended in pure water (HPLG grade, 15 mL) with the help of sonication, and cooled to 0°C. With vigorous stirring, aq NaOH (1 N, 0.182 mL) was added slowly into the suspension from an Eppendorf Pipette. After the addition a sma!l piece of dry ice was added to quench any extra NaOH. The pale yellow solution was then frozen and !yophiilzed to aiford (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yi)- 2-(((2-carboxypropan-2-yl)oxy)imino)acetamido)-3-((1-(2-(1-ethyi-7,8-diriydroxy-4-oxo- 1 ,4-dihydroquinoiine-3-carboxamido)ethyi)pyra

azabicyclo[4.2.0]oct-2-ene-2-carboxylate, Sodium salt (0.164 g, 0.175 mmoi, 96.15% yield) as a light brown solid, LCMS: (M+H)⁺: 813.4. H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.21 - 1.42 (in, 9 H) 2.00 - 2.17 (m, 4 H) 3.32 - 3.60 (m, 8 H) 3.64 - 3.89 (in, 3 H) 3.98 - 4.09 (m, 1 H) 4.52 (d, J=7.07 Hz, 2 H) 5.22 - 5.29 (in, 1 H) 5.71 - 5.80 (m, 1 H) 6.78 - 6.83 (m, 1 H) 6.89 - 6.97 (m, 1 H) 7.47 - 7.54 (m, 1 H) 8.25 - 8.31 (m, 1 H)

Example 47 (6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1-(2-(1 -ethyl-5.6-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2- ene-2-carboxyiate, 2 Sodium salt 18: !θ: 3

Exampte 47a tert-Butyi (3,4-dimethoxyphenyi)carbamate

To a solution of 3,4-dimethoxyaniline (14 g, 91 mmoi) In dry Tetrahydrofuran (THF) (385 mL) was added Boc₂0 (101 mmol) in one portion. The resulting solution was refiuxed for 2 h. After cooling, the solvent was evaporated and the solid residue was suspended in hexane (500 ml), and the mixture was heated to reflux. After cooling down to rt, the white solid was collected by filtration to afford tert-hutyi (3,4-dimethoxyphenyl)carbamate (21 g, 81 mmol, 89 % yield). LCMS: (M+H)⁺: 254.0.

Example 47b 6-((tert-Butoxycarbonyl)amino)-2,3-dimethoxybenzQlc acid

N-butyl!ithium (232 mL, 464 mmol) was added slowly to a solution of tert-butyl (3,4- dimethoxypheny!)carbamate (50 g, 193 mmol) in Tetrahydrofuran (THF) (500 mL) under N₂ at - 20 °C. After being stirred for 2 h at -10 °C to -20 °C, the mixture was cooled to - 78

°C and diluted with Tetrahydrofuran (THF) (500 mL), followed by addition of solid carbon dioxide. The mixture was allowed to warm up to r.t. and was then partitioned between water and Et20. The aqueous layer was acidified with 6N HC! to pH 1-2 and extracted with Et^Q. The organic layer was separated, dried over Na2SC>4 and evaporated. The crude product was purified by recrystailization from /-P^O to afford 6-((tert- butoxycarbonyi)amino)-2,3-dimethoxybenzoic acid (27.9 g, 94 mmol, 48.5 % yield) as a white solid. LCMS: (M+Hf: 319.9.

Example 47c 6-Amino-2,3-dimethoxybenzoic acid, Hydrochloride PU85023

6-((tert-Butoxycarbonyi)amino)-2,3-dimethoxybenzoic acid (39 g, 131 mmol) was treated with HCi in dioxane (4N, 300 mL, 1200 mmol) at r.t. and the mixture was stirred at r.t. for 3 h. LC S indicated completion of the reaction. The solid was collected by filtration, washed with ethyl ether and dried to afford 6-amino-2,3-dimet oxybenzoic acid, Hydrochloride (28.5 g, 120 mmol, 91 % yle!d) as a white solid. LCMS: (M+H)⁺: 198.2.

E am le 47d 5,6-Dimethoxy-l H-benzo[d][1 ,3joxazine-2,4-dione

Bis(trichioromethyl) carbonate (24.89 g, 84 mmol) was added into a solution of 6-amino- 2,3-dlmethoxybenzoic acid, Hydrochloride (28 g, 120 mmol) in Water (1.1 L). The mixture was stirred at r.t. over the weekend. The reaction mixture was filtered, the solid was collected and dried to afford 5,6-dimethoxy-1 H-benzo[d][1 ,3]oxazine-2,4-dione (24 g, 105 mmol, 88 % yield) as a light yellow solid. LCMS: (M+H)⁺: 223.9.

Example 47e 1-Ethyl-5,6-dimetho ,3]oxazine-2,4-dione

To a stirred suspension of sodium Hydride (4.84 g, 8 mmol) In N,N-Dimethy! formamide (DMF) (50 mL) In an ice bath under N2 was added 5,6-dimeihoxy-1 H- benzo[d][1 ,3] oxazine-2,4-dione (22 g, 97 mmol) In Ν,Ν-Dimethylformamide (DMF) (350 mL). After stirring for 15 rnin at r.t., iodoethane (15.81 mL, 193 mmol) in N,N- Dimethylforrnamide (DMF) (50 mL) was added dropwise, and the resulting mixture was allowed to stir at r.t. overnight. The mixture was concentrated in vacuo to one-third of the original volume and then poured into ice-water (800 mL), The precipitate was collected by filtration and recrysta!lized from THF to afford 1-ethyl-5,6-dimethoxy-1 H-benzo[d] [1 ,3]oxazine-2,4-dione (15 g, 54.3 mmol, 56.2 % yield) as a light brown pellet. PU85023

LCMS: ( +H)^÷: 252.0.

Exampte 47f Ethyl 1-ethyl-5,6-dl ,4-dihydroquinoline-3-carboxyiate

Under N2, 1-ethyl-5,6-dimethoxy-1 H-benzo[d][1 ,3]oxazine-2,4-dione (12 g, 43.5 mmol) in

Ν,Ν-Dimethyiformamide (DMF) (250 mL) was added to a solution of (Z)~3-ethoxy~3- oxoprop-1-en-1-oiate, Sodium salt (18.01 g, 130 mmo!) in Ν,Ν-Dimethyiformamide (DMF) (150 mL) with stirring. The resulting solution was stirred at 1 10 ^!,C for 3 h. After cooling down, the mixture was concentrated in vacuo and the orange residue was taken up in water. The aqueous solution was washed with ethyl ether (3 x), acidified to pH-2 with 8 N HCi, and extracted with DCM (4 x). The organic extracts were dried over ^SC^, filtered and concentrated. The residue was purified by normal phase silica gel chromatography (ComblFiash), eluting with MeOH/DCM (0-15%) to afford ethyl 1-ethyi- 5,6-dimethoxy-4-oxo-1 ,4-di ydroquino!ine-3-carboxyiate (4.5 g, 14.74 mmol, 33.9 % yield) as a brown oil that turned into a brown solid after overnight. LCMS: (M+H)⁺: 306.4.

Ethyl 1-ethy!-5,6-dihydroxy-4-oxo-1 ,4-di ydroquinoiine-3-carboxyiate

To a brown solution of ethyl 1-ethy!-5,6-dimethoxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxyiate (6 g, 19.65 mmol) in Dichloromethane (DCM) (100 mL) was added BBr₃ (6.13 mL, 64.8 mmol) dropwise and the mixture was stirred at r.t. overnight. The resulting mixture was carefully poured into EtOH (50 mL) at - 40 °C and the mixture was stirred at r.t. over the weekend. The precipitates were collected by filtration and washed with DCM and MeOH. The filtrate was concentrated a bit, the precipitates were collected and washed by DCM. The solids were combined to afford ethyl 1-ethyl-5,6-dihydroxy-4-oxo- 1 ,4-dihydroquinoiine-3-carboxylate (5 g, 9.02 mmol, 45.9 % yield) as a yellow solid that contained also some 1-eihyl-5,6-dihydroxy-4-oxo-1 ,4-dlhydroquinoline-3-carboxylic acid. This mixture was directly used in the next step without further purification. LCMS: PU85023

(M+H)⁺: 278.4 (ester); 250.3 (acid).

Example 47 Ethyl 1-ethyl-5-hydroxy-6-((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydro quinolirie-3-carboxylate

To a suspension of ethyl 1-ethyl-5,6-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylate (5 g, 18.03 rnmol) and potassium carbonate (8.72 g, 63.1 mmol) in Acetone (120 mL) at r.t. was added 4-methoxybenzyl chloride (7.37 mL, 54.1 mmol) and Kl (0.599 g, 3.81 mmol). The mixture was heated under reflux for 2 days. After cooling, the reaction mixture was filtered and the filtrate was concentrated. The residue was partitioned between DCM and water, and the organic layer was washed with sat. NaHCC^ and brine, dried over ^SG^ and concentrated. The crude product was purified by normal phase automatic silica column chromatography (Combifiash RF), eluting with EtOAc/hexanes (0-100%) to afford ethyl 1-ethyl-5-hydroxy-6-((4-methoxybenzyi)oxy)-4-oxo-1 ,4~dlhydroquinoline~3- carboxyiate (1 .5 g, 3.77 mmol, 20.9% yield) as a light yellow solid that contained some PMB-ester. LCMS: (M+H)⁺: 398.3.

Example 47 i 1-Ethyi~5-hydroxy~8-((4-methoxybenzy!)oxy)~4-oxo~1 ,4~dlhydroqulnoline~3- carboxyilc acid

To a suspension of ethyl 1-ethyl-5-hydroxy-6-((4-methoxybenzyi)oxy)-4-oxo-1 ,4- dihydroquinoline-3-earboxyiate (1.5 g, 3.77 mmol) in a mixture of Methanol (60.0 mL) and Water (20 mL) was added KOH (0.424 g, 7.55 mmol) portionwise. The resulting mixture was stirred under reflux for 1 h. The mixture was cooled down to r.t. and concentrated. Water was added and the mixture was adjusted to ρΗ~2 using 6 N HCI (aq.). The precipitates were collected by filtration and dried in air to afford 1-ethyl-5-hydroxy-6-((4- methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3-carboxyiic acid (1.35 g, 3.85 mmoL 97 % yield) as a light yellow solid. LCMS: (M+H)⁺: 370.2. Exampte 47i 1-Ethyl-5-hydroxy-6-((4-methoxybenzy!)oxy)-4-oxo-N-(2-(pyrro!idin-1 - PU85023 yi)ethyl)-1 ,4-dihydroquinoline-3-carboxamide

To a solution of 1-ethyl-5-hydroxy-6-((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoiine- 3-carboxyiic acid (1.4 g, 3.79 mmo!) in N.N-Dimethyiformamide (DMF) (30 mL) was added HATU (1.729 g, 4.55 mmo!) and DIPEA (1 .655 mL, 9.48 mmol), the resulting mixture was stirred at r.t. for 30 min. Then 2-(pyrrolidin-1 -yi)ethanamine (0.656 mL, 4.17 mmol) was added and the resuiting mixture was stirred at r.t. for 3 h. The reaction mixture was concentrated and the residue was dissolved in DCM and washed with water, sat. NaHCC>3 (aq.) and brine. The organic layer was separated and concentrated, the residue was purified by normal phase silica gel chromatography (CombiFlash), eluting with MeOH/DCM (0-20%) to afford 1-ethyl-5-hydroxy-6-((4-methoxybenzy!)oxy)-4-oxo-N-(2- (pyrrolidin-1 -yi)ethyl)-1 ,4-dihydroquinoiine-3-carboxamide (1 g, 2.148 mmol, 58.7 % yield) as a yellow solid. LCMS: ( +H)⁺: 466.3. ct-2-

(6R7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino)acetamido)-3- ((1-(2-(1-ethyl-5,6-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxamido)et yi)pyrrolidin ium-1 -yl)methyi)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2-carboxyiate was prepared as a slightly brown solid according to the procedure described in Examples 18k~18l, utilizing 1~Ethyl-5~hydroxy-8-((4~methoxybenzyi)oxy)-4~oxo-N-(2~(pyriOildin-1 -yl)ethyl)- 1 ,4-dihydroquinoiine-3-carboxamide (Example 44j) in place of 5-chioro-1~ethyi-6,7-bis((4- methoxybenzyl)oxy)-3-(pyrro!idin-1-yimethyl)quinolin-4(1 H)-one in step 16k.. This product (0.19 g, 0.225 mmol) was suspended in pure water (HPLC grade, 20 mL) with the help of sonication, and cooled to 0°C. With vigorous stirring, aq NaOH (0.2 N, 2.15 mL, 0.43 PU85023 mmo!) was added slowly into the suspension from an Eppendorf Pipette. After the addition a small piece of dry ice was added to quench any extra NaOH. The mixture was then frozen and iyophilized to afford (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)~3-((1-(2~(1 -ethyl-5_!8-dihydroxy-4-oxo-1 ,4- dihydroquinoline-3~carboxamldQ)ethyi)pyrrolidin-1-ium-1 ~yl)methyi)-8-oxo-5~thia-1- azabicyclo[4.2.0]oct-2-ene-2-carboxylate, 2 Sodium salt (0.19 g, 0.192 mmoi) as a slightly brown solid. LCMS: (M+Hf: 843.4. 'H N R (400 MHz, DEUTERIUM OXIDE) δ ppm 1.29 (t, J=7.20 Hz, 3 H) 2.05 - 2.22 (m, 4 H) 2.56 - 2.64 (m, 2 H) 3.41 - 3.61 (m, 6 H) 3.63 - 4.09 (m, 6 H) 4.83 - 4.90 (m, 1 H) 5.23 - 5.29 (m, 1 H} 5.71 - 5.78 (m, 1 H) 6.77 - 6.83 (m, 1 H) 6.88 (s, 1 H) 7.03 - 7.12 (m, 1 H) 8.24 - 8.32 (m, 1 H) oxy)imino)

Example 48a Ethyl 3-(dimethyiamino)-2-(2,4_>5-trif!uorobenzoyi)acryiate

To a solution of t ethylamine (107 mL, 771 mmoi) and ethyl 3-(dimethylamino)acrylate (55.2 mL, 386 mmoi) in toluene (250 mL) was added dropwise a toiuene solution of 2,4,5- trifluorobenzoyl chloride (50 g, 257 mmoi). The mixture was stirred at 90 °C for 3h. LCMS showed completion of reaction. After cooling down, the reaction mixture was washed with water and extracted with EA. The organic layer was dried over Na₂S0₄, filtered and concentrated to afford ethyl 3-(dimethylamino)-2-(2,4,5-trifluorobenzoyl)acryiate (65 g, 216 mmoi, 84 % yield) as a brown oil. LCMS: (M+Hf: 302.1.

Exampte 48b Ethyl 1-(tert-butyl)-6,7-difluoro-4-oxo-1 ,4-dihydroquinoline-3-carboxylate PU85023

To a solution of ethyl 3-(dimethyiamino)-2-(2,4,5-trifluorobenzoyi)acryiate (64 g, 212 mmol) in a mixed solvent of Ethanoi (150 mL)/Diethyl ether (300 mL) was added tert- Butyiamine (44.6 mL, 425 mmol), and the reaction mixture was stirred at rt for 2h. The mixture was concentrated under reduced pressure, the resulting oily residue was dissolved in N,N-Dirnethylformamide (DMF) (250 mL) and potassium carbonate (88 g, 637 mmol) was added. LCMS showed completion of the reaction after being stirred at 100 °C for 2 h. Cold water was added to the mixture, and the precipitates were collected by filtration and dried to afford ethyl 1-(tert-butyl)-6,7-difiuoro-4-oxo-1 ,4-dihydroquinoline- 3-carboxyiate (60 g, 194 mmol, 91 % yield) as pale yellow solid. LCMS: (M+H)⁺: 310.1

Exarnpfe 48c 1-(tert-Butyl)-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxyilc acid

A mixture of ethyl 1 -(tert-butyi)-6,7-difluoro-4-oxo-1 ,4-dihydroquinoline-3-carboxyiate (50 g, 162 mmol), KOH (136 g, 2425 mmol) and 4-methoxybenzyl alcohol (201 ml, 1617 mmol) was heated at 80 ^t!C under N₂ for 8h. LCMS showed completion of reaction. The solution was adjusted to pH 2, extracted with EA and washed with water. The orgainc phase was concentrated in vacuo and the crude product was triturated In Et₂0 to afford 1- (tert-butyl)-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3-carboxylic acid (61 g, 1 18 mmol, 72.9 % yield) as pale yellow solid. LCMS: (M+H)⁺: 518.3.

Exampte 48d 1~(tert-Butyi)-6J-bis((4~methoxybenzyl)oxy)-3-(pyrroildin-1 -ylmethyi) quinolln-4(1 H)-one

This compound was prepared according to the procedure described in Examples 44g-44i, utilizing 1-(tert-butyi)-6_>7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxy!lc acid (Exampie 48c) in place of 1 -eihyl-7,8-bis((4-methoxybenzy!)oxy)-4-oxo- 1 ,4-dihydroquinoiine-3-carboxylic acid in step 44g. LCMS: (M+H)^÷: 557.4.

Exampie 48e (6R,7R)-7-((Z)-2-(2-aminothiazol-4-y!)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-(tert-buty!)-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-y!)rriethyl) pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2 3oct-2-ene-2-carboxylate, Sodium salt

The compound was prepared according to the procedure described In Examples 18k-18! and 17, utilizing 1-(tert-butyl)-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1 - ylmethyl)quinolin-4(1 H)-one (Example 48d) in place of 5-chloro-1-ethyl-6,7-bis((4- methoxybenzyl)oxy)-3-(pyrrolidin-1 -yimethyl)quinolin-4(1 H)-one in step 16k. LCMS: (M+H)⁺: 814.3. ^!H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1 .69 - 1 .78 (m, 9 H) 1.95 - 2.1 1 (m, 4 H) 2.75 (s, 2 H) 3.22 - 3.47 (m, 6 H) 3.84 - 3.93 (m, 1 H) 3.95 - 4.05 (m, 1 H) 4.30 - 4.45 (m, 2 H) 4.84 - 4.91 (m, 1 H) 5.24 - 5.31 (m, 1 H) 5.71 (d, J=4.80 Hz, 1 H) 6.90 (s, 1 H) 7.51 (s, 1 H) 7.54 (s, 1 H) 8.22 y) line-3- 03oct-2-

Exampie 49a 1-(tert-Butyl)-6, -oxo-N-(2-(pyrrolidin-1 -yi) ethyl)- 1 ,4-dihydroquino!ine-3-

PU85023

This compound was prepared according to the procedures described in Example 45a, utilizing 1-(tert-butyi)-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxy!ic acid (Example 48c) In place of 1-ethyi~7,8-bis((4~methoxybenzyl)oxy)-4~oxo- 1 ,4-dihydroquinoiine-3-carboxylic acid. LCMS: (M+H : 614.5.

Example 49b (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-(2-(1-(tert-butyl)-6_>7-dihydroxy-4-oxo-1 _l4-dihydroquinoiine-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate, Sodium salt

The compound was prepared according to the procedure described in Examples 16k-16l and 17, utilizing 1-(tert-butyi)-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1 - yl)ethy!)-1 ,4-dlhydroquino!ine-3-carboxamide (Example 49a) in p!ace of 5~ch!oro~1 -ethyl- 6,7-bis((4-methoxybenzyi)oxy)-3-(pyrroiidin-1 -ylmethyl)quinoiin-4(1 H)-one in step 16k. LCMS: (M+Hf: 871.4. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.88 (s, 9 H) 2.03 - 2.21 (m, 4 H) 2.58 - 2.67 (m, 2 H) 3.34 - 3,60 (m, 7 H) 3.64 - 3,76 (m, 2 H) 3,78 - 3.90 (m, 2 H) 4.06 (d, J=14.15 Hz, 1 H) 4.78 - 4.87 (m, 1 H) 5.23 (d, .7=5.05 Hz, 1 H) 5.71 (d, J=4.80 Hz, 1 H) 6,83 (s, 1 H) 7.39 (s, 1 H) 7.42 - 7.46 (m, 1 H) 8.67 - 8.74 (m, 1 H) Example 50 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-cyciopropy!-6,7-dihydroxy-4-oxo-1 ,4-dihydroquino!in-3-yi) methyl) pyrrol idin-1-ium-1-yi)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate, Sodium salt

Example 50a Ethyl 1 -cyclopropyi-6,7-difiuoro-4-oxo-1 ,4-dihydroquinolirie-3-carboxy!ate

A solution of ethyl 3-(dimethyiamino)-2-(2,4_!5-trif!uorobenzoyl)acrylate (Example 48a) (75 g, 249 rnmol) in a mixture of Ethanol (100 mL)/Diethyi ether (200 mL) was added to cyciopropanamine (34,5 mL, 498 mrnol). After 2 h of stirring at rt, LC S showed complete disappearance of starting material. The reaction mixture was evaporated under reduced pressure, the oily residue was dissolved in Ν,Ν-Dimethyiformamide (DMF) (250 mL) and potassium carbonate (103 g, 747 mrnol) was then added. The reaction mixture was stirred at 100 °C for 2 h, and LCMS showed completion of reaction. Cold water was added to the mixture. The precipitates were collected by filtration and dried to afford ethyl 1-cyciopropyl-6,7-d!fluoro-4-oxo-1 ,4-dihydroquinoi!ne-3-carboxylate (71 g, 242 mrnol, 97 % yield) as a pale yellow solid. LCMS : [M+Hf : 294.1.

Example 50b 1-Cyciopropyl-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxyiic acid

A mixture of ethyl 1 -cyciopropy!-6,7-difluoro-4~oxo-1 ,4-dihydroquinoline~3~carboxylate (29 g, 99 mrnol), KOH (83 g, 1483 mmoi) and MeOH (400 ml, 9887 mrnol) was heated at 130 °C under N₂ for 48 h. LCMS showed completion of reaction. The solution was adjusted to ρΗ 2, concentrated in vacuo and washed with water to afford 1-cyclopropyl- 6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxyl!c acid (20 g, 89.1 mrnol, 89.9 % yield) as pale yellow solid. The crude product was used in next step without further purification. LCMS : [M+H]⁺ : 290.1 .

Exam le 50c 1-cyclopropyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylic acid

To a solution of 1-cyclopropyi-6,7-dirnethoxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxylic acid !θ: 3

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To a solution of 1-cyciopropyl~6J-dihydroxy-4-oxo-1 ,4-dihydroqulnoline~3-carboxyilc acid (8.2 g, 31.4 mmol) In Ν,Ν-Dimethylformamide (D F) (150 mL) was added potassium carbonate (21.69 g, 157 mmol), followed by 1-(chioromethyi)-4~methoxybenzene (17.04 mL, 126 mmol). The reaction mixture was stirred at 50 ^CiC overnight. LCMS indicated completion of the reaction. Water was added and the mixture was stirred at r.t for 15mins. The yellow precipitates were collected by filtration and washed with water to afford 4~ methoxybenzy! 1 -cyclopropyi-6_l7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoiine- 3-carboxyiate (14 g, 22.52 mmol, 71.7 % yield) as a yellow solid that contained 1- cyclopropyi-6,7-bis((4-methoxybenzy!)oxy)-4-oxo-1 ,4-dihydroquinoiine-3-carboxyiic acid (Example 50e). The crude mixture was used in next step without purification.

Exarnpfe 50e 1-Cyclopropyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxyilc acid

To a suspension of 4-methoxybenzy! 1-cyclopropyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo- 1 ,4-dihydroquinoiine-3-carboxylate (14 g, 22.52 mmol) in a mixture of Methanol (100 mL) and Water (50.0 mL) was added KOH (2.53 g, 45.0 mmol) portionwise. The resulting mixture was stirred at 90 °C for 2 h. LCMS indicated completion of the reaction. The reaction mixture was cooled down to r.t. and concentrated, and was then diluted with PU85023 water and adjusted pH to 1 using 2 N HCI (aq.). The precipitates were collected by filtration and dried to afford 1-cyclopropyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroqulnoline~3-carboxyilc acid (1 1 g, 21.93 mmol, 97 % yield) as pale yellow solid. LCMS : [M+Hf : 502.3.

This compound was prepared according to the procedures described In Examples 44g- 44i, utilizing 1-Cyciopropyi-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxyilc acid (Example 50e) in place of 1-ethyl-7,8-bis((4-rriethoxybenzyl)oxy)-4-oxo- 1 ,4-dihydroquinoline-3-carboxylic acid in step 44g. LCMS: (M÷H)^÷: 541 .4.

Exampte SOq (6R,7R)-7-((Z)-2-(2-aminot iazol-4-yl)-2-(((S)-1 _!2-diGarboxyethoxy)imino) acetamido)-3-((1-((1-cyciopropyl-6 -dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-yl)methyi) pyrrol idin-1-ium-1-y!)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2,03oct-2-ene-2-carboxylate, Sodium salt

The compound was prepared according to the procedure described In Examples 16k-18! and 17, utilizing 1-cyclopropyl-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1- ylmethyl)quinolin-4(1 H)-one (Example 50f) in place of 5-chloro-1-ethyi-6,7-bis((4- methoxybenzyl)oxy)-3-(pyrrolidin-1 -ylmethyl)quinolin-4(1 H)-one in step 16k. LCMS: (M+Hf: 798.3. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 0.93 - 1 .06 (m, 2 H) 1.15 - 1.29 (m, 2 H) 1.93 - 2.10 (m, 4 H) 2.65 - 2.76 (m, 2 H) 3.23 - 3.57 (m, 6 H) 3.83 - 3.95 (m, 1 H) 3.96 - 4.08 (m, 1 H) 4.24 - 4.43 (m, 2 H) 4.62 - 4.67 (m, 1 H) 4.83 - 4.91 (m, 1 H) 5.23 - 5.33 (m, 1 H) 5.69 - 5.77 (m, 1 H) 6.92 (s, 1 H) 7.43 - 7.47 (m, 1 H) 7.48 - 7.53 (m, 1 H) 8.17 - 8.24 (m, 1 H)

E ample 51 (6R.7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)-1 ,2-diearboxyethoxy) imino)acetamido)-3-((1-((1-cyclopropyl- 6.7-dihydroxy-4-oxo-1.4-dihydroquinolin-3-yl) PU85023 methyl)pyrrolidin-1 -ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- earboxy!ate, Sodium salt

The compound was prepared according to the procedure described in Examples 211-21 m, utilizing 1-cyclopropyi-6 J-bis((4-methoxybenzyl)oxy)-3-(pyrro!idin-1 -yimethyl)quinoiin- 4(1 H)-one (Example 50f) in place of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-3- (pyrrolidin-1 -yimethyi)quinolin-4(1 H)-one in step 211.. LCMS: (M+Hf: 832.0. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 0.89 - 1 .04 (m, 2 H) 1.1 1 - 1.25 (m, 2 H) 1 .88 - 2.09 (m, 4H) 2.75 (s, 2 H) 3.25 - 3.50 (m, 8 H) 3.82 - 3.92 (m, 1 H) 3.94 - 4.04 (m, 1 H) 4.20 - 4.39 (m, 2 H) 4.61 - 4.67 (m, 1 H) 4.82 - 4.92 (m, 1 H) 5.21 - 5.30 (m, 1 H) 5.66 - 5.75 (m, 1 H) 7.33 (s, 1 H) 7.35 - 7.42 (m, 1 H) 8.07 - 8.16 (m, 1 H).

Example 52 (6R_!7R)-7-((Z)-2-(2-amino-5-criiorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetarnido)-3-((1 -((1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolirv y!)methyl)pyrro!idin-1 -ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oGt-2-ene-2- carboxyiate, Sodium salt

The compound was prepared according to the two-step sequence of Examples 22f-22g, utilizing 1-cyclopropyl-6 ,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1-yimethyl)quinolin- 4(1 H)-one (Example 50f) in place of 5-chloro-1-ethyl-6,7-bis((4-methoxybenzyl)oxy)-3- (pyrrolidin-1 -ylmethyi)quinolin-4(1 H)-one in step 22f. LCMS: (M+H)⁺: 801 .9. H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 0.90 - 1 .03 (m, 2 H) 1.13 - 1.26 (m, 2 H) 1.39 (d, J=6.57 Hz, 6 H) 1.88 - 2.09 (m, 4 H) 3.21 - 3.52 (m, 6 H) 3.85 - 4.04 (m, 2 H) 4.20 - 4.40 (m, 2 H) 4.62 - 4.88 (m, 1 H) 5.24 - 5.34 (m, 1 H) 5.77 (d, J=5.05 Hz, 1 H) 7.35 (s, 2 H) 8.12 (s, 1 H).

Example 53 (6R JR)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyetrioxy)irriirio) acetamido)-3-((1 -(2-(1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2-carboxylate, 2 Sodium salt PU85023

Example 53a 1-Cyclopropyi-6,7-bis((4-methoxybenzy!)oxy)-4-oxo-N-(2-(pyrro!idin-1- y!)ethyl)-1 ,4-dihydroquinoline-3-carboxamide

This compound was prepared according to the procedures described In Example 45a, utilizing 1-Cyciopropyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxyllc acid (Example 50e) in place of 1-ethyl-7,8-bis((4-methoxybenzyl)oxy)-4-oxo- 1 ,4-dihydroquinoiine-3-carboxylic acid. LCMS: (M+H)⁺: 598.4.

.Example 53b (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1 -(2-(1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene~2-carboxylate, 2 Sodium salt

This compound was prepared according to the procedures described in Example 47k, utilizing 1-cyciopropyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1-yl)ethyi)-1 ,4-di hydro quinoline-3-carboxamide (Example 53a) in place of 1-Ethyl-5-hydroxy-6-((4-methoxy benzyi)oxy)-4-oxo-N-(2-(pyrroiidin-1-yl)ethyl)-1 ,4-dihydroquinoline-3-carboxamide. LCMS: (M+H)⁺: 855.4. ¹H NMR (400 MHz, DEUTERIUM OXIDE) ij ppm 0.95 (d, J=3.03 Hz, 2 H) 1.14 - 1.26 (m, 2 H) 2.13 (d, J=3.03 Hz, 4 H) 2.55 - 2.66 (m, 2 H) 3.30 - 3.61 (m, 8 H) 3.63 - 3.76 (m, 2 H) 3.77 - 3.90 (m, 2 H) 4.03 - 4.14 (m, 1 H) 4.82 - 4.89 (m, 1 H) 5.23 - 5.29 (m, 1 H) 5.75 (d, J=4.80 Hz, 1 H) 6.83 (s, 1 H) 7.14 - 7.23 (m, 2 H) 8.27 - 8.33 (m, 1 H)

Exampte 54 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) PU85023 acetamido)-3-((1-((5-c

yi)methyl)pyrroiidin-1-i .0]oct-2-ene-2- earboxy!ate, 2 Sodium

Example 54a 1-Cyclopropyi-6,7-dimet oxy-5-nitro-4-oxo-1 ,4-dihydroquino!ine-3- carboxylic acid

1-Cyclopropyl-6,7-dimethoxy-4-oxo-1 ,4-dihydroquinoline-3-carboxylic acid (Example 50b) (20 g, 69.1 mmo!) was dissolved in sulfuric acid (44.2 ml, 830 mmol) with stirring. The thick dark solution was cooled to 0 °C and potassium nitrate (10.48 g, 104 mmol) was added In small portions, while the temperature was maintained below 10 °C by use of an Ice-water bath. After the addition, the mixture was kept under 10 °C for 1 h, and then was allowed to warm up to r.t. and stirred overnight. LCMS showed completion of reaction. The mixture was poured into ice-water (300 mL). A yellow solid precipitated out and was collected by filtration, washed with water and ethano!, and dried in vacuo to afford 1- cyciopropyl-6,7-dimethoxy-5-nitro~4-oxo~1 ,4~dlhydroquinoline~3-carboxylic acid (14 g, 41 .9 mmol, 60.6 % yield) as a pale yellow solid. LCMS : [M+Hf : 335.1.

E am le 54b 1-Cyclopropyl-6,7-dimethoxy-5-nitro-4-oxo-1 ,4-dihydroquinoline-3- earboxylic acid

Under N2, 1-cyclopropyi-6,7-dimethoxy-5-n!tro-4-oxo-1 ,4-dihydroquinoiine-3-carboxylic acid (10 g, 29.9 mmol) was treated with sodium sulfide nonahydrate (71.9 g, 299 mmol) In a mixture of Ethanol (100 mL) and Water (100 mL) for 2 h at 100 °C. LCMS showed completion of reaction. After cooling down, the mixture was poured into cold water and the solution was adjusted to pH 2. The yellow precipitate was collected by filtration, washed with water and dried to afford 5-amino-1-cyclopropy!-6,7-dimethoxy-4-oxo-1 ,4- dihydroquinoiine-3-carboxylic acid (8.9 g, 29.2 mmo!, 98 % yield) as a yellow solid.

LCMS : [M+Hf : 305.1 .

Exampie 54c 5-Chloro-1 -cyclopropyl~8-hydroxy-7-methoxy~4-oxo~1 ,4~rJihydroqulnoline-3- carboxyile acid

To a pale brown suspension of 5-amino-1 -cyclopropy!-6,7-dimethoxy-4-oxo-1 ,4- dihydroquinoiine-3-carboxylic acid (9 g, 29.6 mmol) In con. HCi (60 mL) stirred at 0 °C was added dropwise a co!d solution of sodium nitrite (2.143 g, 31 .1 mmo!) in Water (20.00 mL). Stirring continued at 0 °C for 1 h. LCMS indicated completion of reaction. HCI (80 mL, 2633 mmol) was added and the mixture was heated at 95 °C for 6 h. LCMS Indicated completion of reaction. The reaction mixture was cooled down to rt and then poured into water. The precipitates were collected by filtration and dried to afford 5- c ioro-1-cyciopropy!-6-hydroxy-7-methoxy-4-oxo-1 ,4-di ydroquino!ine-3-carboxylic acid (7.4 g, 23.89 mmol, 81 % yield) as a pale yellow solid. LCMS : [M+Hf : 310.1.

Exampie 54d S-Chloro-l-cyclopropyi-BJ-dihydroxy^-oxo-l ^-dihydroquinoiine-S- carboxyilc acid

To a solution of 5-chioro-1-cyciopropyl-6-hydroxy-7-methoxy-4-oxo-1 ,4-dihydroquinoline- 3-carboxyiic acid (8.5 g, 27.4 mmol) In Dichioromethane (DCM) (100 mL) was added BBr₃ (6.49 mL, 68.6 mmol) at -78 °C. The mixture was allowed to warm up to rt, and stirred at rt overnight. LCMS Indicated completion of the reaction. The mixture was diluted with MeOH and concentrated to dryness. This procedure was repeated several times to afford 5-chioro-1-cyciopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxyiic acid (7.8 g, 26.4 mmol, 96 % yield) as yellow solid. LCMS: [ +Hj⁺: 296.1.

Exampie 54e 4-Methoxybenzyl 5-chioro-1 -cyclopropy!-6,7-bis((4-methoxybenzyl)oxy)-4- oxo- 1 ,4-d i hyd roq u i nol i n e-3-ca rboxy late PU85023

To a solution of 5-chioro-1-cyciopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxyiic acid (1 1 g, 37.2 mmoi) in Ν,Ν-Dimet yiformamide (DMF) (120 mL) was added potassium carbonate (25.7 g, 188 mmo!), followed by 1-(chioromethyi)-4- met oxybenzene (20.20 mL, 149 mmoi). The reaction mixture was stirred at 50 °C for 5 h. LCMS indicated completion of the reaction. Water was added and the mixture was stirred at r.t for 5 mins. The yellow precipitates were collected by filtration and washed with water to afford 4-methoxybenzyi 5-chioro-1-cyciopropyi-6,7-bis((4-methoxy benzyl) oxy)-4-oxo-1 ,4-dihydroquinoi!ne-3-carboxylate (20 g, 30.5 mmoi, 82 % yield) as a yeliow solid. LCMS: [M+Hf : 856.5.

Exam le 54 5-Chloro-1 -cyclopropyi-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4- dihydroquinoiine-3-carboxylic acid

To a suspension of 4-methoxybenzyi 5-chloro-1 -cyclopropy!-6,7-bis((4-methoxy benzyl) oxy)-4-oxo-1 ,4-dihydroquinoiine-3-carboxylate (18 g, 27.4 mmoi) in a mixture of

Methanol (120 mL) and Water (60.0 mL) was added potassium hydroxide (3.08 g, 54.9 mmoi) portionwise. The resulting mixture was stirred at 90 °C for 3 h. LCMS indicated completion of the reaction. The reaction mixture was cooled down to r.t. and

concentrated, and was then diluted with water and adjusted pH to 1 using 2 N HCI (aq.). The precipitates were collected by filtration and dried to afford 5-chloro-1-cyclopropyl-6,7- bis((4-met oxybenzyi)oxy)-4-oxo-1 ,4-dihydroquinoline-3-carboxyiic acid (12.6 g, 23.51 mmoi, 86 % yield) as pale yellow solid. LCMS: [M+Hf : 536.3. Example 54g 5-Chioro-1-cyclopropyl~6,7~bis((4-methoxybenzyl)oxy)-3~(pyrrolidin-1 - y!methyl)quinoliri-4(1 H)-one

PU85023

This compound was prepared according to the procedures described in Examples 44g- 441, utilizing 5-chioro-1 -cycioprapyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydro quinollne-3-carboxylic acid (Example 54f) in place of 1-ethy!-7,8-bis((4-methoxybenzyi) oxy)-4-oxo-1 ,4-di ydroquino!ine-3-Garboxylic acid in step 44g. LC S: (M+H)⁺: 575.2. iin-

This compound was prepared according to the procedures described In Example 47k, utilizing 5-chloro-1-cyciopropyl-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1 -ylmethyi) quinolin-4(1 H)-one (Example 54g) in place of 1-Ethyl-5-hydroxy-6-((4-methoxybenzyi) oxy)-4-oxo-N-(2-(pyrrolidin-1 -yi)ethyl)-1 ,4-dihydroquinoiine-3-carboxamide. LCMS: (M+Hf: 832.1. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 0.87 - 1 .03 (m, 2 H) 1.13 - 1.26 (m, 2 H) 1.98 - 2.1 1 (m, 4 H) 2.57 - 2.69 (m, 2 H) 3.27 - 3.53 (m, 6 H) 3.83 - 3.94 (m, 1 H) 3.96 - 4.09 (m, 1 H) 4.16 - 4.34 (m, 2 H) 4.73 - 4.77 (m, 1 H) 4.85 - 4.92 (m, 1 H) 5.26 - 5.35 (m, 1 H) 5.74 (s, 1 H) 6.90 (s, 1 H) 7.29 - 7.38 (m, 1 H) 8.05 - 8.14 (m, 1 H). Exarnpfe 55 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yi)oxy)imino) acetamido)-3-((1-((5-chloro-1 -cyclopropyl-6.7-dihydroxy-4-oxo-1.4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiaie, Sodium salt

The compound was prepared according to the procedure described in Examples 19b~19c and 20, utilizing 5-ch!oro-1 -cyc!opropyl~6,7~bis((4~meihoxybenzyi)oxy)-3~(pyrrolidln-1 - yimethy!)quinolin-4(1 H)-one (Example 54g) in place of 5-chloro-1-ethyl-6,7-bis((4- methoxybenzyl)oxy)-3-(pyrroiidin-1-yimethyl)quinolin-4(1 H)-one in step 19b. LCMS: (M+H)⁺: 802.0. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 0.87 - 1 .03 (m, 2 H) 1.09 - 1.24 (m, 2 H) 1.39 (d, J=6.82 Hz, 6 H) 1.93 - 2.12 (m, 4 H) 3.37 (br. s., 6 H) 3.90 (br. s., 2 H) 4.14 - 4.38 (m, 2 H) 5.27 - 5.39 (m, 1 H) 5.73 - 5.81 (m, 1 H) 6.86 (s, 1 H) 7.24 - 7.34 (m, 1 H) 8.02 - 8.13 (m, 1 H). )oxy)irriino) uinoiine-3- ,03oct-2-

Example 58a 5-Chioro-1 -cyciopropyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2- (pyrrolidin-1 -yi)ethyi)-1 ,4-dihyd

This compound was prepared according to the procedures described in Example 45a, utilizing 5-chioro-1-cyciopropyi-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydro quinoline-3-carboxylic acid (Example 54f) in place of 1-ethy!-7,8-bis((4-methoxybenzyi) oxy)~4-oxo~1 ,4~dlhydroqulnoline-3-carboxyilc acid. LC S: (M+H)⁺: 632.5.

Exarnpfe 58b (6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-3-((1-(2-(5-chloro-1 -cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-d!hydro quinoline-3-carboxamido)ethyi)pyrrolidin-1^^

[4.2.0]oct-2-ene-2-carboxyiate, Sodium sa!t

The compound was prepared according to the procedure described In Examples 19b-19c and 20, utilizing 5-chioro-1 -cyciopropyl-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2- (pyrrolidin-1 -y!)ethyi)-1 ,4-dlhydroqulnoline-3-carboxamide (Example 56a) In place of 5- PU85023 chloro-1-ethy!-67-bis((4-methoxybenzyi)oxy)-3-(pyro^

in step 19b. LCMS: (M+H)⁺: 859.0. ¹H N R (400 MHz, DEUTERIUM OXIDE) δ ppm 0.92 - 1.07 (m, 2 H) 1.16 - 1.28 (m, 2 H) 1.34 (s, 6 H) 2.04 - 2.20 (m, 4 H) 3.37 - 3.82 (m, 8 H) 3,68 - 3.81 (m, 1 H) 3,82 - 3.95 (m, 2 H) 4.00 - 4.12 (m, 1 H) 4.66 (m, 1 H) 5.24 - 5.31 (m, 1 H) 5.73 - 5.80 (m, 1 H) 6.83 - 6.89 (m, 1 H) 7.17 - 7,26 (m, 1 H) 8,43 - 8.52 (m, 1 H).

Example 57 (6RJR)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imlno) acetamido)-3-((1-((6 J-dihydroxy-1-methyi-4-oxo-1 ,4-dihydroquinolin-3-yi)meth pyrrolidin-l-ium-l-yiJmethy -S-oxo-S-thia-l -azabicyclo^^.OjocW-ene^-carboxylate, 2 Sodium salt

Example 57a 6 J-Bis((4-methoxybenzyl)oxy)-1-methyl-3-(pyrrolidin-1-yimethyl)quinolin- 4(1 H)-one

This compound was prepared according to the procedures described in Examples 50a~ 50f, utilizing methanarnine in place of cyciopropanamine in Example 50a. LCMS: (M+H)⁺: 515.3.

Example 57b (6RJR)-7-((Z)-2-(2-arninothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6 J-dihydroxy-1-methyl-4-oxo-1 ,4-dihydroquinolin-3-yi)methyl) pyrrol!din-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate, 2 Sodium salt

This compound was prepared according to the procedures described In Example 47k, utilizing 6 -bis((4-methoxybenzyi)oxy)-1-methyi-3-(pyrrolidin-1-ylmethyi)quinolin-4(1 H)- one (Example 57a) in place of 1-Ethyl-5-hydroxy-6-((4-methoxybenzyl)oxy)-4-oxo-N-(2- PU85023

(pyrrolidin-1 -yl)ethyl)-1 ,4-dihydroquino!ine-3-carboxamide. LCMS: (M+H)^÷: 772.3.

NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.85 - 2.08 (m, 4 H) 2.52 - 2.61 (m, 2 H) 3.23 - 3.45 (m, 8 H) 3.75 (s, 3 H) 3.80 - 3.91 (m, 1 H) 3.96 - 4.05 (m, 1 H) 4.21 - 4.31 (m, 1 H) 4.32 - 4.42 (m, 1 H) 4.83 (s, 1 H) 5.22 - 5.28 (m, 1 H) 5.88 - 5.75 (m, 1 H) 6.87 (s, 2 H) 7,41 (s, 1 H) 7.99 - 8.06 (m, 1 H).

Example 58 (6RJR)-7-((Z)-2-(2-aminothiazo!~4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imlno) acetamido)-3-((1-((6,7-dihydroxy-1-isopropy!-4-oxo-1 ,4-dihydroquinolin-3-y!)met yl) pyrrolidin-l-ium-l-yiJmethy -S-oxo-S-thia-l -azabicyclo^^.OjocW-ene^-carboxylate, 2 Sodium sait

Example 58a 1-lsopropyi-6_!7-bis((4-methoxybenzyi)oxy)-3-(pyrroiidin-1-ylmet yl)quinolin-

4(1 H)-one

This compound was prepared according to the procedures described in Examples 5Qa-50f, utilizing propan-2-arnine in place of cyclopropanamine in Example 50a. LCMS: (M+H)⁺: 543

Example 58b (6R JR)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6,7-dihydroxy-1-isopropy!-4-oxo-1 ,4-dihydroquinolin-3-y!) methyl) pyrrolidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate, 2 Sodium salt

This compound was prepared according to the procedures described in Exampie 47k, utilizing 1~lsQpropyl~8,7~bls((4-methoxybenzyi)oxy)-3-(pyrroiidin-1-yimethyl)quinQlin~ 4(1 H)-one (Exampie 58a) in place of 1 ~Ethyi-5~hydroxy-8-((4~methoxybenzyl)oxy)-4~oxo~ N-(2-(pyrro!idin-1 -yl)ethyl)-1 ,4-dihydroquinoiine-3-carboxamide, LCMS: (M+H)^*: 800.1 . 'H PU85023

NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.39 - 1.49 (m, 8 H) 1.92 - 2.10 (m, 4 H) 2.54 - 2.62 (m, 2 H) 3.20 - 3.48 (m, 8 H) 3.84 - 3.93 (m, 1 H) 3.95 - 4.06 (m, 1 H) 4.29 - 4.42 (m, 2 H) 4.84 (none, 2 H) 5.23 - 5.30 (m, 1 H) 5.89 - 5.76 (m, 1 H) 6.85 - 6.91 (m, 1 H) 7, 10 - 7.17 (m, 1 H) 7,41 - 7.48 (m, 1 H) 8.09 - 8.15 (m, 1 H)

Example 59 1 -(((6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-y!)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-t ia-1-azabicycio[4.2 ]oct-2-en-3- y!)met yl)-1-(2-(5-chioro-1 -ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethy!)pyrroi!din-1-ium, Trifluoroaceiic acid salt

E am le 59a 1-(((6R,7R)-2-((benzhydryloxy)carbonyl)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-

((4-methoxybenzyl)oxy)-1 ^-dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyl)amino)-

5-chlorothiazol-4-yi)acetamido)-8-oxo-5-t ia-1 -azabicyclo[4.2 ]oct-2-en-3-yl)methyi)-1-

(2-(5-c loro-1-ethyl-6,7-bis((4-methoxybenzyi)oxy)-4-oxo-1 ,4-di ydroquinoline-3- carboxamido)ethyi)pyrroi!din-1-ium

The compound was prepared according to the procedure described in Examples 2 1, utilizing 5-c !oro-1-ethy!-6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidiri-1-yi)ethyl)- 1 ,4-dihydroquinoiine-3-carboxamide (Example 27a) in place of 5-ch!oro-1 -ethy!-6,7-bis((4- methoxybenzyl)oxy)-3-(pyrroiidin-1-yimethyl)quino!in-4(1 H)-one. LCMS: (M+H)⁺: 1595.5.

Example 59b 1-(((6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2 ]oct-2-en-3- yl)rnethyl)-1-(2-(5-chioro-1-ethyi-6_>7-dihydroxy-4-oxo-1 _>4-dihydroquinoline-3- carboxamido)ethyl)pyrrol!din-1-ium, Trifiuoroacetic acid salt PU85023

To a solution of 1~(((8RJR)-2-((benzhydryioxy)carbonyl)-7-((Z)~2-((((S)-4-(tert-butoxy)-1 ~ ((4~methoxybeRzyi)oxy)-1 ,4-dioxobutan-2-y!)oxy)lmino)-2-(2~((teri-buioxycarb^

5-chlorothiazoi-4-yl)acetamido)-8-oxo-5-thia~1 ~azabicycio[4.2 joci-2-en~3-yl)^

(2-(5-chioro-1-ethyi-6J-bis((4-methoxybeR^

carboxamldo)ethy!)pyrrolidin-1-ium (0.172 g, 0.108 mmol) in Dichloromethane (DCM) (3 mL) under N2 at 0°C was added anlso!e (0.1 18 mL, 1.078 mmol), followed by TFA (0.5 mL, 6.49 mmol). The mixture was warmed up to rt and stirred at rt over 24 h. LCMS Indicated completion of the deproteetion. Diisopropyl ether (6 mL) and water (1 mL) were then added. The mixture was stirred for 10 min and the solvents were decanted away from the solid. The solid was triturated twice with diisopropyl ether (5 mL) and dried under high vacuum to afford 1-(((6R,7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)- 1 ,2-dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2- en-3-yl)methyi)-1-(2-(5-ch!oro-1-ethy!-6 ,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethy!)pyrro!!din-1-ium, Trifiuoroacetic acid salt (91 mg, 0.044 mmol, 41.1 % yield) as a slightly brown solid. LCMS: (M+H)⁺: 913.0. H NMR (400 MHz, METHANOL- d4) 5 ppm 1 .46 - 1 ,55 (m, 3 H) 2.19 - 2.34 (m, 4 H) 2.95 - 3.04 (m, 2 H) 3.41 - 4.47 (m, 14 H) 4,98 - 5.02 (m, 1 H) 5.28 - 5.33 (m, 1 H) 5.85 - 5.93 (m, 1 H) 7.1 - 7.14 (m, 1 H) 8.68 - 8,75 (m, 1 H)

Exaropte 60 1 -(((6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-y!)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3- y!)rnethyi)-1-(2-(1 -ethyl-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethy!)pyrroiidin-1-ium, Trifiuoroacetic acid salt

Example 60a 1-Ethyi-8-fiuoro-6,7-bis((4~methoxybenzyi)oxy)-4~Qxo-N-(2-(pyriOildin~1 - y!)ethyl)-1 ,4-dihydroquinoiine-3-carboxamide PU85023

This compound was prepared according to the procedures described in Example 45a, utilizing 1-ethyi-8-fiuoro-6 ,7-bis((4-methoxybenzyl)oxy)-4-oxo-1 ,4-dihydroquinoline-3- carboxyiic acid (Example 15i) in place of 1-ethyl-7,8-bis((4-methoxybenzy!)oxy)-4-oxo- 1 ,4~rJlhydroqulnoline-3-carboxyilc acid. LCMS: (M+H)⁺: 604.5.

Exampte 60b 1-(((6R,7R)-7-((Z)-2-(2-amino-5-chloroihiazol-4-yl)-2~(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2 ]oct-2-en-3- y!)methyi)-1~(2-(1 ~ethyi-8-fiuoro~6,7~dihydroxy-4~oxo~1 ,4-dihydroquinoline-3- carboxamido)ethyi)pyrroiidin-1-ium, Trifiuoroacetic acid salt

The compound was prepared according to the procedure described in Examples 59a- 59b, utilizing 1-ethy!-8-f!uoro-6J-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrro!idin-1- yl)ethyl)-1 ,4-dihydroquinoline-3-carboxamide (Example 80a) in place of 5-chloro-1 -ethyl- 6,7-bis((4-methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1-yi)ethyl)-1 ,4-dihydroquinoiine-3- carboxamlde. LCMS: (M+H)⁺: 895.4. ¹H NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.32 - 1.43 (m, 3 H) 2.04 - 2.30 (m, 4 H) 2.70 - 2.89 (m, 2 H) 3.31 - 3.77 (m, 8 H) 4.13 - 4.20 (m, 1 H) 4.28 - 4.36 (m, 2 H) 4.41 - 4.49 (m, 1 H) 4.81 - 4.90 (m, 2 H) 5.00 (s, 1 H) 5,35 - 5.42 (m, 1 H) 5,51 - 5.58 (m, 1 H) 6.81 - 6.89 (m, 1 H) 7.25 - 7.34 (m, 1 H) 8.33 - 8,40 (m, 1 H)

Exaropte 61 1 -(((6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-y!)-2-(((S)-1 ,2- dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3- y!)methyl)-1-((1-ethyi-8-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquino!in-3- y!)methyl)pyrro!idin-1-ium, Trifiuoroacetic acid salt PU85023

The compound was prepared according to the procedure desc ibed in Examples 59a- 59b, utilizing 1-ethyl-8-fiuoro-6,7-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1 - ylmethyi)quinolin-4(1 H)-one (Example 151) in place of 5-chioro-1-ethyi-6,7-bis((4- methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidin-1 -yi)ethyi)-1 ,4-dihydroqLiinoline-3- carboxam.de. LCMS: (M+H)^÷: 838.2. Ή N R (400 MHz, METHANGL- 4) δ ppm1.43 (t, J=6.82 Hz, 3 H) 2.10 - 2.31 (m, 4 H) 2.95 - 3.04 (m, 2 H) 3.47 - 3.73 (m, 6 H) 4.26 - 4.67 (m, 6 H) 5.12 - 5.20 (m, 1 H) 5.35 - 5.39 (m, 1 H) 5.86 - 5.94 (m, 1 H) 6.98 - 7.05 (m, 1 H) 8.17 - 8.20 (m, 1 H).

Example 82 (6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-6,7-dihydroxy-1-methyl-4-oxo-1 ,4-dihydroquino!in-3~ y!)methyl)pyiTo!idin-1~ium-1~yl)methyl)-8-oxo-5-thia-1~azabicycio[4.2.0joci-2-ene~2- carboxyiate, 2 Sodium salt

y!methy!)quinolin-4(1 H)-one

This compound was prepared according to the procedures for the preparation of 5-chloro- 1-cyciopropyl-6 J-bis((4-methoxybenzyl)oxy)-3-(pyrrolidin-1 -yimethyl)quinolin-4(1 H)-on as described in Examples 50a-50b and 54a-54g, utilizing methanamine in place of cyclopropanamine in Example 50a. LCMS: (M+H)⁺: 549.3.

-Example 62 (6R,7R)-7-((Z)-2-(2-aminoth!azol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)!mino PU85023 roquinolin-3- .0]oct-2-ene-2-

This compound was prepared according to the procedures described in Example 47k, utilizing 5-ch!oro-6_l7-bis((4-methoxybenzy!)oxy)-1-methyl-3-(pyrroiidin-1- y!methy!)quinoiln-4(1 H)-one (Example 62a) in place of 1-Ethyi-5-hydroxy-6~((4~ methoxybenzyl)oxy)-4-oxo-N-(2-(pyrrolidir!-1 -y!)ethyi)-1 ,4-dihydroquinoiine-3- carboxamlde. LC S: (M+H)⁺: 895.4. Ή NMR (400 MHz, DEUTERIUM OXIDE) δ ppm 1.88 - 2.07 (m, 4 H) 2.50 - 2.88 (m, 2 H) 3.18 - 3.49 (m, 6 H) 3.85 (s, 3 H) 3.83 - 3.93 (m, 1 H) 3.94 - 4.06 (m, 1 H) 4.1 1 - 4.21 (m, 1 H) 4.22 - 4.32 (m, 1 H) 4.82 - 4.88 (m, 1 H) 5.24 - 5.32 (m, 1 H) 5.72 (d, J=4.80 Hz, 1 H) 6.63 - 6.74 (m, 1 H) 6.85 - 6.92 (m, 1 H) 7.88 - 7.98 (m, 1 H)

It Is to be understood that the Invention is not limited to the embodiments illustrated hereinabove and the right is reserved to the illustrated embodiments and all modifications coming within the scope of the following claims.

The various references to journals, patents, and other publications which are cited herein comprise the state of the art and are incorporated herein by reference as though fully set forth.

Claims

PU85023

What is claimed is:

1. A compound of Formula (

wherein:

X is N, or C-R^a;

R¹ and R² each are hydrogen, (Chalky!, or (CH₂)p-C(0)OR^b;

R³ and R⁴ each are hydrogen, OH or OR^c;

wherein:

R³ is hydrogen or halogen;

R^B or R^c each is H, (C _-6)-alkyl, an alkali meia! or negative charge;

A is R⁵ or -NR^dC(0)R⁵

wherein R^d Is H or (C^-alkyl

R^¾ is an optionally saturated or unsaturated monocyclic heterocyclic ring or an optionally saturated or unsaturated bi-cyclic or fused heterocyclic ring;

wherein:

each fused heterocyclic ring optionaiiy includes carbocyclic rings or heterocyclic rings of up to four heteroatoms;

R⁵ optionally Is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0). -CN, -N0₂,-halogen, - straight or branched C_1-6 a!kyi- straight or branched C_1-6 aloalkyi, C₃.₆-cycloalkyl, - straight or branched d. 6 straight or branched aikoxy, -OC(0)OH, -OG(0}R^e, -C(0)OR^f, -0(CH₂)_yOR^g, - NR^!1R' , ~S0₂R^j, -S(CH₂)_qR^k, -NR'C(0)R^m, aryl or heteroaryi;

wherein:

hetero atoms are selected from oxygen, nitrogen or sulphur;

carbocyclic rings or heterocyclic rings for each fused heterocyclic ring systems include non-aromatic rings or aromatic rings; monocyclic heterocyclic rings or fused heterocyclic rings include substituted aromatic and non-aromatics;

each R^e, R^f, R⁹, R^h, R^!, R^j , R^k, R¹ ,or R^m each is selected from H, C-i alkyl, C haioaikyl, C alkoxy;

each aryl or heieroaryi as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - N0₂, -halogen, Ci_₆ -alkyl, -haioaikyl, Ci_-6 -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)_y,OR^p, -NR^qR^r , -S0₂R^s, -SiCHzJy-R', - NR^uC(0)R^v;

wherein:

R", R°, R^p, R^q, R R^s, R^} ,R^U, or R^v each are selected from C _-6 alkyl, C_1-6 -haioaikyl, or Ci_-s-alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

2. A compound of Formula (ill):

wherein:

X Is N, or C-R^a;

R¹ and R² each are hydrogen, (C₁.₆)alkyi, or (CH₂)p-C(0)OR^b:

R³ and R⁴ each are hydrogen, OH or OR^c;

wherein:

R^a is hydrogen or halogen;

R^B or R^c each is H, (C-i_₆)-alkyi, an a!kail metal or negative charge;

A is R⁵ or ~NR^dC(0)R⁵

wherein:

R^Q is H or (Ci-eJ-alkyl

R⁵ is a rnonocyciic 3 to 7 membered heterocyclic ring or a bicyclic 10 membered heterocyclic ring; PU85023 wherein:

each 3 to 7 membered heterocyclic ring contains up to four heteroatoms or each blcyclic 10 membered heterocyclic ring contains up to four heteroatoms;

wherein:

hetero atoms are selected from oxygen, nitrogen or sulphur; carbocyclic rings or heterocyclic rings for each 10 membered heterocyclic ring systems contain non-aromatic rings or aromatic rings;

R5 optionally is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-haiogen, - straight or branched C _-6 alky!,- straight or branched C _-6 aloalkyl. C:3__B-eycioaikyl, - straight or branched Ci_ β straight or branched alkoxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -0(CH₂)_yOR⁹, - NR^hR' , -S0₂R^j, -S(CH₂)_qR^k, -NR'C(0)R^m, aryl or heteroaryl;

wherein:

each aryl or heteroaryl as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - N0₂, -halogen, Ci_₆ -aikyl, Ci^ -haioaikyi, C_1-6 -alkoxy, -OC(0)OH, -

OC(0)R^R, -C(0)OR°, -0(CH₂)_yX)R^p, -NR^qR^r , -S0₂R^s, -SiGH^R¹, - NR^uC(0)R^v;

wherein:

Rⁿ, R°, R^p, R^q, R^r, R^s, R¹ ,R^U, or R^v each as defined above are selected from Ci-₆ aikyl, Ci.₆ -haloalkyl, or Ci._B~aikoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

3, The compound of Formula (IV):

PU85023

wherein:

X is C-R^a;

R¹ and R² each are hydrogen, (C,..₆)aikyl, or (CH₂)p~C(0)OR^b;

R³ and R⁴ each are hydrogen, OH or 0R^C;

wherein:

R is hydrogen or halogen;

R or R^c each is H, (Ci_₆)-alkyl, an alkali metal or negative charge;

A is R⁵ or ~NR^dC(0)R⁵

wherein R^d is H or (C _-6)-alkyl;

R⁵ is an optionally saturated or unsaturated monocyclic heterocyclic ring or an optional!y saturated or unsaturated bi-cyc!ic or fused heterocyclic ring;

wherein:

each fused heterocyclic ring optionally includes carbocyciic rings or heterocyclic rings of up 4 heteroatoms;

R5 optionally is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0),-CN, -NQ₂,-halogen, - straight or branched Ci.₆ aikyl,- straight or branched Ci_₆ haloalky!, C₃.₆cyc!oa!kyi, - straight or branched Ci_₆ straight or branched a!koxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -0(CH_z)_yOR⁹, - NR^hR' , -8Q₂R^j, -S(CH₂)_qR^k, ~NRⁱC(0)R^rri, aryi or heteroaryl;

wherein:

hetero atoms are selected from oxygen, nitrogen or sulphur;

each R^e, R^f, R⁹, R^h, R^;, R^j , R^k, R¹ ,or R^m each is selected from H, Ci.₆ alkyl, C .₆ haloalky!, C-|..₆ alkoxy; PU85023 each aryi or heteroaryl as defined above is optionally substituted with one or more of the following substituenis selected from H, -OH, -CN, - N0₂, -halogen, C _-6 -aiky!, C^ -haloaikyi, C-i.e -alkoxy, -OC(0)OH , - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)_y.OR^p, -NR^qR^r , -S0₂R^s,

- NR^uC(0)R^v;

wherein:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each are selected from Ci_6 alkyi, C-i_₆ -ha!oa!kyi, or Ci_-6-alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

4. The compound of Formula (IV) according to claim 3, wherein X is C-R^a and R³ is hydrogen .

A compound of Formula (V):

wherein :

X is C-R^a;

R¹ and R² each are hydrogen , (C,..₆)alkyi, or (CH₂)p~C(0)QR^b;

R³ and R⁴ each are hydrogen , OH or OR^c;

wherein :

R^a is hydrogen or halogen :

R or R^c each is H , (C-i -ej-aikyl, an alkali metal or negative charge;

A Is R⁵ or ~NR^dC(0)R⁵

wherein :

R^d is H or (Ci_₆)-alkyi

R⁵ is a monocyclic 3 to 7 membered heterocyclic ring or a bicyclic 10 membered heterocyclic ring ;

wherein : PU85023 each 3 to 7 membered heterocyclic ring contains up to four heteroatoms; each bicyciic 10 membered heterocyclic ring contains up to four heteroatoms; wherein:

each heteroatom is selected from nitrogen, oxygen or sulfur;

each 10 membered heterocyclic ring optionally contains carbocyclic rings or heterocyclic rings;

wherein:

carbocyclic rings or heterocyclic rings for each 10

R5 optionally is substituted by one or more of the following substituenis selected from -H, -OH, Oxo (=0), -CN, -N0₂,-haiogen, - straight or branched C _-6 alkyl,- straight or branched Ci_₆ aloalky!, C₃.₆-cycioaikyl,- straight or branched C>.₆ straight or branched a!koxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -0(CH₂)_yOR⁹, - NR^hR' , -S0₂R^j, -S(CH₂)_qR^k, -NR^;C(0)R^rr!, aryl or heteroary!;

wherein:

each R^e, R^f, R⁹, R^h, R^;, R^j , R^k, R¹ ,or R^m as defined above is selected from H, Ci .₆ alkyl, Ci-₆ aloalkyl, C a!koxy;

each ary! or heteroaryl as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, -

N0₂, -halogen, C_1-e -aikyl, Ci^ - aioaikyi, Ci_₆ -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CHzVOR^p, -NR^qR^r , -S0₂R^s, -S(CH₂)_y-R¹ _I - NR^uC(0)R^v;

wherein:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each as defined above are selected from Ci_₆ alkyl, Ci_₆ -haioaikyl, or C-i..₆-aikoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof. 6, A compound of Formula (Vi): PU85023

wherein:

X is C-R^a;

R¹ and R² each are hydrogen, (C,_-6)aikyl, or (CH₂)p-C(0)OR^b;

R³ and R⁴ each are hydrogen, OH or OR^c;

wherein:

R^s is hydrogen or halogen;

R" or R^c each is H, (C^-alkyl, an alkali metal or negative charge;

A I

wherein:

R₆, R₇, R₈, R₉, R₉., R₉.., R₉>», R₁₀₎ Rn , R₁₂ or R₁₇ is H, straight or hranched(Ci__B)-a!kyl or C₃.₆-cycioaikyl;

each A optionally further is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,~ha!ogen, - straight or branched C _-6 aikyl,- straight or branched Ci_₆ haioaikyi, C₃.₆-cycloalkyl, - straight or branched C _-6 PU85023 straight or branched aikoxy, -OC(0)OH , -OC(0)R^e, -C(0)OR^f, -0(CH₂)_yOR^g, -NR^t5R^! , - S0₂R^j, ~S(CH₂)_qR\ -NR'C(0)R^m _> ary! or heieroaryl;

where:

each R^e, R^f, R⁹, R^h, R^!, R^j , R^k, R¹ ,or R^m as defined above is selected from H, Ci_₆ aiky!, C-i..₆ haloalkyi, C .₆ aikoxy;

each aryl or heieroaryl as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, N0₂, -halogen, Ci_₆ -aikyl, C^ -ha!oaikyi, Ci_-6 -aikoxy, -OC(0)OH , - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)_y R^p, -NR^qR^r , -SO_zR^s, -S(CH₂)_y-R^t, - NR^uC(0)R^v;

where:

R", R°, R^p, R^q, R^r, R^s, R^} ,R^U, or R^v each as defined above are selected from aikyl, C^ -haioaikyi, or C i.₆-a!koxy; and n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

7. The compound of Formula according to claim 8, wherein A is

PU85023

wherein:

R_6> R₇, Re, RQ, RIO, R I or R₁₂, R_13l R_M, Ris, Rie, R17, Ris, is, R20, R21 , R22, or R₂₃ is H, straight or branched(Ci_₆)-alky! or C₃.₆-cyc!oa!kyi

each A optionally further is substituted by one or more of the following

substituents selected from -H, -OH, Oxo (=0), -CN, -NQ₂,-ha!ogen, - straight or branched C_h alky!,- straight or branched C-..₆ ha!oa!kyl, C₃.₆-cycloalkyl, - straight or branched Ci_₆ straight or branched alkoxy, -OC(0)OH, -OC(0)R^e, -C(0)OR^f, -0(CH₂)_yOR⁹, -NR^hR^; , - 8Q₂R^j, -S(CH₂)_qR^k _! -NR'C(0)R^m, aryl or heteroaryi;

where:

each R^e, R^f, R⁹, R^h, R^;, R^j , R^k, R¹ ,or R^m as defined above is selected from H, Ci .₆ aikyl, Ci-₆ aloalkyl, C- .₆ alkoxy;

each aryl or heteroaryi as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - N0₂, -halogen, Ci_₆ -aikyl, d-e -haloaikyi, Ci_₆ -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)y.OR^p, -NR^qR^r , -S0₂R^s,

- PU85023

NR^uC(0)R^v;

where:

Rⁿ, R°_; R^p, R^q, R^r, R^E, R* ,R^U, or R^v each as defined above are selected from C-| .₆ aikyl, Ci.₆ -haloalkyi, or Ci..₆-alkoxy; and n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

8, The compound of Formula (VI) according to claim 8, wherein:

Ra is hydrogen;

R¹ and R each are (C_1-S)alkyl;

R³ and R⁴ each are OH or OR⁰;

wherein:

R⁸ or R ^'2 is H, (Ci..₆)-alkyl or (C₃..₆)-cycloalky!;

n is 1 ; or

a pharmaceutically acceptable salt thereof

9. A compound of Formula (VIH):

wherein:

X Is C-R^a;

R¹ and R² each are hydrogen, (C<.₆)alkyi, or (CH₂)p-C(0)OR^b

R³ and R⁴ each are hydrogen, OH or OR^c; PU85023 wherein:

R^a is hydrogen or halogen;

Rⁿ or R^c each is H, (Ci_₆)-alkyl, an alkali metal or negative charge;

A Is

wherein:

Is H, straight or branched(Ci_₆)-alkyl or C₃.₆-cyc!oa!kyl.

each A optionally further is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-haiogen, ~ straight or branched C-i aikyi,- straight or branched C .₆ ha!oa!kyi, C₃.₆-cycloalky!, - straight or branched Ci_₆ straight or branched aikoxy, -OC(0)OH, -OC(0)R^s, ~C(0)OR^f, ~0(CH₂)_yOR⁹, -NR^"R , - S0₂R^j, -S(CH₂)_qR\ -NR^Q .R"¹, aryl or heteroaryl;

where:

each R^e, R^f, R⁹, R , R^!, R^j , R^k, R¹ ,or R^m as defined above is selected from H, Ci aikyl, Ci-e haloalkyl, C^e aikoxy;

each aryl or heteroaryl as defined above is optionally substituted PU85023 with one or more of the following substituents selected from H, -OH, -CN, N0₂, -halogen, C _-6 -aiky!, C^ -haloaikyi, C-i.e -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)y-OR^p, -NR^qR^r , -S0₂R^s,

- NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each as defined above are selected from Ci_₆ aikyl, C_1-6 -haloalkyl, or Ci_₆-alkoxy; and n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

10. The compound of Formula (VII) according to claim 9, wherein X is C-R^a and R^a is hydrogen.

A compound of Formula (V!il):

wherein:

R¹ and R² each are hydrogen, (Chalky!, or (CH₂)p-C(0)OR^b;

R³ and R⁴ each are hydrogen, OH or OR^c;

wherein:

R^B or R^c each is H, (C _-6)-alkyl, an alkali metal or negative charge;

A is R⁵ or ~NR^dC(0)R⁵

Wherein:

R^d is H or (C_1-e)-alkyl

R^¾ is an optionally saturated or unsaturated monocyclic heterocyclic ring or an optionally saturated or unsaturated bi-cydic or fused heterocyclic ring:

wherein:

each fused heterocyclic ring optionally includes carbocyclic rings or PU85023 heterocyciic rings of up to four eteroatoms;

R5 optionally Is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, ~NQ₂,~halogen, - straight or branched Ci_₆ aikyi,- straight or branched C _-6 ha!oa!ky!, C₃.₆~cycioaikyl,- straight or branched C _s straight or branched aikoxy, -OC(0)OH, ~OC(0)R^e, -C(0)OR^f, -0(CH₂)_yOR⁹, - NR^hR' , -8Q₂R^j, -S(CH_z)_qR^k, -NRⁱC(0)R^rri, ary! or heteroaryi;

wherein:

hetero atoms are selected from oxygen, nitrogen or sulphur;

each R^e, R^f, R^g, R^h, R', R^j , R^K, R¹ ,or R^m each is selected from H, C alkyl, C>..₆ haloalkyl, Ci.₆ aikoxy;

each aryl or heteroaryi as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, ~CN, ~ N0₂,~halogen, -aikyi, C.,.₆ -haloalkyl, -aikoxy, -OC(0)OH, - OC(0)R^R, -C(0)OR°, -0(CH₂)_y.OR^p, -NR^qR^r , -S0₂R^s, -SiCHs -R', - NR^uC(0)R^v;

wherein:

Rⁿ, R°, R^p, R^q, R^r, R^s, R¹ ,R^U, or R^v each are selected from Ci aikyi, Ci_₆ -haloalkyl, or Ci-e-alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or

a pharmaceutically acceptable salt thereof.

12. A compound of Formula (iX):

wherein:

R¹ and R² each are hydrogen, (C,..₆)alkyi, or (CH₂)p-C(0)OR^b; PU85023

R³ and R⁴ each are hydrogen, OH or OR⁰;

wherein:

Rⁿ or R^c each is H, (Ci_₆)-alkyl, an a!ka!i metal or negative charge: A is R⁵ or -NR^dC(0)R⁵

where R^d is H or (C< _₆)-a!kyl

R⁵ is a monocyclic 3 to 7 membered heterocyclic ring or a bicyclic 10 membered

heterocyclic ring;

wherein:

each 3 to 7 membered heterocyclic ring contains up to four heteroatoms; each bicyclic 10 membered heterocyclic ring contains up to four heteroatoms; where:

heteroatoms are selected from oyygen, nitrogen or sulphur;

each 10 membered heterocyclic ring optionally contains carbocyc!ic rings or heterocyclic rings;

wherein:

carbocydic rings or heterocyclic rings for each 10

R5 optionally is substituted by one or more of the following substituents selected from -H, -OH, Oxo (=0), -CN, -N0₂,-halogen, - straight or branched C _-6 alkyi- straight or branched C^e haloalky!, C₃,₆-cycloalkyl, - straight or branched Ci_ 6 straight or branched aikoxy, -OC(0)OH, -OG(0}R^e, -C(0)QR^f, -0(CH₂)_yOR^g, - NR^hR' , -S0₂R^j, -S(CH₂)_qR\ -NR^!CfO)R^m, aryi or heteroaryi;

wherein:

each R^e, R R⁹, R' R^>, R^j , R^k, R¹ ,or R^rn as defined above is selected from H, Ci_₆ aikyl, Ci_₆ haloalkyl, C>..₆ a!koxy;

each aryi or heteroaryi as defined above is optionally substituted with one or more of the following substituents selected from H, -OH, -CN, - N0₂,-halogen, C-,.₆ -aikyl, C-i ₆ -haloa!ky!, C.,.₆ -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH_z)_y.OR^p, -NR^qR^r , -S0₂R^s,

-

NR^uC(0)R^v;

wherein:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each as defined above are selected from Ci_₆ aikyl, C _-6 -haioaikyl, or Ci_₆-alkoxy;

n, m, o, p, q or y each are 0 or an integer from 1 to 5; or PU85023 a pharmaceutically acceptable salt thereof.

13. The compound according to c!aim 12, wherein A is :

wherein :

is H, straight or branched(C-;_₆)-alkyl or C₃.₆-cyc!oa!kyl .

each A optionally further is substituted by one or more of the following substituents selected from ~H, -OH, Oxo (=0), -CN , -N02,-haiogen, ~ straight or branched C-i 6 aikyi,- straight or branched Ci.₆ haloalkyi, C₃.₆~cycloalkyl , - straight or branched Ci_₆ straight or branched aikoxy, -OC(0)OH , -OC(0)R^s, -C(0)OR^f, -0(CH₂)_yOR⁹, -NR^"R , - S0₂R^j, -S(CH₂)_qR\ -NR^Q .R"¹, aryi or heteroaryl;

where:

each R^e, R^f, R⁹, R , R^!, R^j , R^k, R¹ ,or R^m as defined above is selected from H, Ci_6 aikyl, C-i-K haloalky!, C^e aikoxy;

each aryi or heteroaryl as defined above is optionally substituted PU85023 with one or more of the following substituents selected from H, -OH, -CN, - N0₂, -halogen, C _-6 -aiky!, C^ -haloaikyi, C-i.e -alkoxy, -OC(0)OH, - OC(0)Rⁿ, -C(0)OR°, -0(CH₂)y-OR^p, -NR^qR^r , -S0₂R^s,

- NR^uC(0)R^v;

where:

Rⁿ, R°, R^p, R^q, R^r, R^s, R* ,R^U, or R^v each as defined above are selected from Ci_₆ aikyl, C_1-6 -haloalkyl, or Ci_₆-alkoxy; and n, m, o, p, q or y each are 0 or an integer from 1 to 5.

14. The compound according to claim 12, wherein:

R¹ and R each are (C_1-S)alkyl;

R³ and R⁴ each are OH or OR⁰;

wherein:

R° is H, (Ci_₆)-alkyl, an alkali metal or negative charge;

A .s

R⁸ or R '² is H, (C_1-6)-alkyl or (C₃-₆)-cycloalkyi;

n is 1 ; or

a pharmaceutically acceptable salt thereof

15. A compound which is :

1-(((6R,7R)-7-((Z)-2-((((S)-4-(tert-butoxy)-1-((4-met oxybenzyl)oxy)-1 ,4- dioxobutan-2-yl)oxy)imino)-2-(2-((tert-butoxycarbonyi)amino)thiazol-4-yi)acet^

(((4-meihoxyben2y!)oxy)carbonyl)-8-oxo-54hia-1-azablcyclo[4.2.0]oct-2-en-3-yi)methyl)-1 - (2-(5-chloro-6 -bis((4-methoxybenzyi)oxy)-1 -oxo-3,4-dihydroisoquinolin-2(1 H)- yi)eihyl)pyrrolidin-1 ~lum;

(6R,7R)-7-((Z)~2-(2-aminoihiazol-4-yl)-2-(((S)~1 ,2~dicarboxyeihoxy)imino) aceiamido)-3-((1-(2-(5-hydroxy-1~meihyi-4-oxo~1 ,4~dlhydropyridine-2~carboxamldo) ethyl)pyrrolidin-1-ium-1-yl)methy!)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)~7-((Z)~2-(2~aminothiazol~4-yl)~2-(((S)-1 ,2~dicarboxyethoxy)imino) PU85023 acetamido)-3-((1 -(2-(7-hydroxy-1 ,8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- yi)ethyl)pyrrolidin-1 -ium-1-yi)methyi)-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-ene-2- carboxy!ate:

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-y!)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1 -(2-(7-hydroxy-1 ,8-dioxo-3,4-dihydro-1 H-pyrido[1 ,2-a]pyrazin-2(8H)- y!)ethyl)pyrrolidin-1 -ium-1-yi)methy!)-8-oxo-5-t ia-1-azabicycio[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamido)-3-((1 -((3-hydroxy-1 -methyl-4-oxo-1.4-dihydroquinolin-6-yl)methyl)pyrrolidin-1- ium-1 -yi)methyl)-8-oxo-5-thia-1-azabicycio[4.2.0]od-2-ene-2-carboxyiate;

1-(((6R_>7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yi)rnethyl)-1-(2-(3-hydroxy- 1-methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)ethyi)pyrrolidin-1-ium;

1-(((6R,7R)-7-((Z)-2-(2-aminot iazoi-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3-yi)methyl)-1-(2-(3- hydroxy-1 -methyi-4-oxo-1 ,4-dihydroquinoline-6-carboxamido)ethyi)pyrrolidin-1-ium;

1-(((6R,7R)-7-((E)-3-(2-aminothiazol-5-y!)-3-(((R)-1 ,2-dicarboxyethoxy)imino)-2- oxopropyi)-2-carboxy-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-en-3-yi)methyl)-1-((1-ethy!- 6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methy!)pyrro!idin-1-ium;

1-(((6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((R)-1 ,2-dicarboxyethoxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyc!o[4.2.0]oct-2-en-3-yi)methy!)-1-((1-ethyi- 6,7-dihydroxy-4-oxo-1 ,4-dihydroquino!in-3-yl)methyi)pyrroiidin-1-ium;

1-(((6R.7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yi)methyl)-1-((1-ethyi- 6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi)methyl)pyrrolidin-1-ium;

1-(((6RJR)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy)im

acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2-en-3-yi)methyl)-1-(2-(1-ethyl- 6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxamido)ethyl)pyrrolidin-1 -ium;

1-(((6R_!7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropar!-2-y!)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2-en-3-yi)methyl)-1-(2-(1 -ethyl- 6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3-carboxamido)ethyl)pyrroiidin-1 -ium;

1 -(((6R_>7R)-7-((E)-3-(2-aminothiazol-5-yi)-3-(((R)-1 _>2-dicarboxyethoxy) imino)-2- oxopropyi)-2-carboxy-8-oxo-5-thia-1 -azabicyc!o[4.2.03oct-2-en-3-yi)methyl)-1-((1-ethyi- 6,7-dihydroxy-4-oxo1 ,4-dihydrocinnolin-3-yi)methyl)piperidin-1 -ium;

1-(((6R_l7R)-7-((Z)-2-(2-aminothiazoi-4-yl)-2-(((2-carboxypropan-2- PU85023 yi)oxy)imino)acetamido)-2-carboxy-8-oxo-5-th^

1-(2-(3- ydroxy-1 -methyi-4-oxo-1 ,4-dihydroquinoline-8-carboxamido)ethyi)pyrrolidin-1- ium;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((1-ethy!-8-f!uoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-d!hydroxy-4-oxo-1 ,4-d!hydroqu!nolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R₎7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((5-chloro-1 -et yl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxyiatopropan-2-yl)oxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroiidin-1-ium-1-yl)met yi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxy!ate:

(6R,7R)-7-((Z)-2-(2-amino-5-chlorot iazol-4-yl)-2-(((S)-1 _!2-dicarboxyet oxy)imino) acetamido)-3-((1-((5-chloro-1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R_>7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((2-carboxypropan-2-yi)oxy) imino)acetamido)-3-((1 -((5-chloro-1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate; PU85023

(6R7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((1-ethyi-6 -di ydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyl)pyrrolidiri- 1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetamido)-3-((1-((1-et yl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazoi-4-yl)-2-(((2-carboxypropan-2- yl)oxy)imino)acetamido)-3-((1-((1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-d!hydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-et yi-6 J~dihydroxy-4-oxo-1 ,4-dihydroqulno!in-3-yl)meihy!)pyrro!ldln- 1-ium-1-yi)methyl)-8-oxo-5-t ia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazoM-yl)-2-(((S)-1 ,2 licarboxyethoxy)imino) acetamido)-3-((1-(2-(5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)et yl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2~carboxyiate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazo!-4-yl)-2-(((2-carboxypropan-2- yi)oxy)imino)acetamido)-3-((1 -((1-et yl-6,7-dihydroxy-4-oxo-1 ^-dihydrocinnolin-S- yi)methyl)pyrroiidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1.2-dicarboxyethoxy) imino) acetamido)-3-((1 -((1-ethyi-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yi) methyi)pyrroiidin- 1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino) acetamido)-3-((1-(2-(1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3- Garboxamido)ethy!)pyrroiidin-1-ium-1-y!)rriethyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene~2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-(2-(5-chloro-1-ethyl-6_>7-dihydroxy-4-oxo-1 _>4-dihydrocinnoiine-3- carboxamido)ethy!)pyrroiidin-1-ium-2-y!ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo

[4.2.0]oct-2-ene-2-carboxyiate;

1 -(((6R_>7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((2-carboxypropan-2-yl)oxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-en-3-yl)methyi)-1-(2-(5- PU85023 chioro-1-ethyi-6 -di ydroxy-4-oxo-1 ,4-dihydrocinnoline-3-carboxa!Tiido)ethan-1-ylium-1- yi)pyrroiidin-1 -ium;

(6R7R)-7-((Z)-2-(2-amino-5-chlorothiazoM-yl)-2-(((S)-1 ,2 licarboxyethoxy) imino)acetamido)-3-((1 -(2-(5-chloro-1-et yl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3- carboxamido)ethy!)pyrroiidin-1-ium-2-y!ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo

[4.2.0]oct-2-ene-2-carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 _l2-dicarboxyethoxy) imino) acetamido)-3-((1-((5-chloro-1-ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 _>2-dicarboxyethoxy) imino)acetamido)-3-((1 -((5-chloro-1-ethyi-6.7-dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1-((1-ethyl-5-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoliri-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-((1-ethyi-5-fiuoro-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiin-3-yl)methyi) pyrrol idin-1-ium-1-y!)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2,03oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-fluoLiro-1 -ethyi-6.7-dihydroxy-4-oxo-1.4-dihydroquinolin-3-yl) methyl)pyrrolidin-1 -ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate:

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy) imino) acetamido)-3-((1 -(2-(1-ethyl-5-fluoro-6,7-dihydroxy-4-oxo-1 ,4-dihydrOquinoiirie-3- carboxamido)ethy!)pyrroi!din-1-ium-1-y!)rriethyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene~2-carboxylate;

(6R,7R)-7-((Z)-2-(2-amlno-5-chiQrothlazoi-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy) imino)acetamido)-3-((1-((1-ethyl-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)methyl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamidoJ-S-iil-iil-ethyi-B-fluoro^^-dihydroxy^-oxo-l ^-dihydroquinoiin-S-y methyi) PU85023 pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R R)-7-((Z)-2-(2-arninothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-ethyi-6-fiuoro-7,8-dihydroxy-4-oxo-1 ,4-dihydroquirioiin-3-yl) methyl) pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-erie-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1 -(2-(1-ethyl-6-fluoro-7,8-dihydroxy-4-oxo-1 ,4-dihydrOquino!ine-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2,03oct-2- ene-2-carboxyiate;

(6R₎7R)-7-((Z)-2-(2-arninothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-ethyi-7 -di ydroxy-4-oxo-1 ,4-dih^

1-ium-1-yl)rnethyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy) imino) acetamido)-3-((1-(2-(1-ethyi-7,8-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate;

(6R7R)-7-((Z)-2-(2-amiriothiazol-4-yi)-2-(((2-carboxypropari-2-yl)oxy)imino) acetamido)-3-((1-(2-(1-ethy!-7,8-dihydroxy-4-oxo-1 ,4-dihydroquir!o!irie-3-carboxamido) ethyl)pyrrolidin-1-ium-1-yl)methy!)-8-oxo-5-thia-1 -azabicycio[4.2.0]oct-2-ene-2- carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)

!mino)acetamido)-3-((1-(2-(1 -ethyl-5,6-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrrolidin-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2- ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-(tert-butyi)-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- yi)methyl)pyrroHdin-1-ium-1-yl)metriyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate:

(6R,7R)-7-((Z)-2-(2-amiriothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)

imino)acetamido)-3-((1-(2-(1-(tert-butyi)-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethy!)pyrroiidin-1-ium-1-y!)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate;

(6R7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamido)-3-((1-((1-cyciopropy!-6,7-dihydroxy-4-oxo-1 ,4-dihydroquino!in-3-yl) methy!) pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-ene-2-carboxylate;

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 _>2-dicarboxyethoxy) PU85023 imino)acetamido)-3-((1 -((1 -cyciopropyl-6,7-dihydroxy-4-oxo-1 ,4~dihydroqulnoiin~3-y!) methyl)pyrrolidin-1 -ium-1 -yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxy!ate:

(6R,7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((2-carboxypropan-2- y!)oxy)imino)acetarnido)-3-((1 -((1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinolin-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamido)-3-((1 -(2-(1-cyclopropyl-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoline-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)methyl)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2- ene-2-carboxyiate;

(6R₎7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-chloro-1 -cyclopropyl-6_!7-dihydroxy-4-oxo-1 ,4-dihydroquinoliri-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-arninothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-((5-chloro-1 -cyclopropyl-6_!7-dihydroxy-4-oxo-1 _!4-dihydroquinolin-3- yl)methyl)pyrrolidin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((2-carboxypropan-2-yl)oxy)imino) acetamido)-3-((1-(2-(5-chloro-1 -cyclopropyi-6,7-dihydroxy-4-oxo-1 ,4-dihydroquinoiine-3- carboxamido)ethyl)pyrroiidin-1-ium-1-yl)rnethyl)-8-oxo-5-thia-1-azabicyclo[4.2.03oct-2- ene-2-carboxyiate;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yl)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6,7-dihydroxy-1-methyi-4-oxo-1 ,4-dihydroquinolin-3-yl)methyl) pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.0]oct-2-ene-2-carboxylate;

(6R R)-7-((Z)-2-(2-arninothiazol-4-yi)-2-(((S)-1 ,2-dicarboxyethoxy)imino) acetamido)-3-((1-((6,7-dihydroxy-1-isopropy!-4-oxo-1 ,4-dihydroquinoliri-3-yi)methyl) pyrrol idin-1-ium-1-yl)methyl)-8-oxo-5-thia-1 -azabicyclo[4.2.03oct-2-erie-2-carboxylate;

1-(((6R_!7R)-7-((Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(((S)-1 ,2-diGarboxyethoxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 3od-2-en-3-yl)methyi)-1-(2-(5- chioro-1-ethy!-6 J-dihydroxy-4-oxo-1 ,4-dihydroquino!ine-3-carboxamido)ethyl)pyrroNdin-1 - ium;

1-(((6R_>7R)-7-((Z)-2-(2-amino-5-chiorothiazol-4-yl)-2-(((S)-1 _l2-dicarboxyethoxy) imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2 3oct-2-en-3-yl)methyi)-1-(2-(1- PU85023 ethy ί-8-f ! uoro-6 ,7-d i hyd roxy-4-oxo- 1 ,4-d i hyd roq u i no! I n e-3-ea rboxam ido)et hyi)pyrroiidin-1- ium;

1-(((6RJR)-7-((Z)-2-(2-am.no-5^h!oroth!azol-4-y!)-2-(((S)-1 ,2-di

imino)acetamido)-2-carboxy-8-oxo-5-t ia-1-azabicyclo[4.2 ]oGt-2-en-3-yl)methyi)-1-((1 - ethyl-8-fluoro-6 -dihydroxy-4-oxo-1 ,4-dihydroquinolin-3-y!)methyl)pyrro!idin-1 -iurri;

(6R,7R)-7-((Z)-2-(2-aminothiazol-4-yi)-2-(((S)-1 _l2-dicarboxyethoxy)imino) acetamido)-3-((1-((5-c loro-6,7-dihydroxy-1-met yl-4-oxo-1 ,4-dihydroquino!in-3- y!)met yl)pyrro!idin-1-ium-1-yl)methyi)-8-oxo-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2- carboxylate; or

a pharmaceutically acceptable salt thereof.

18. The compound according to claim 15, wherein the pharmaceutically acceptable salt is a sodium salt, di-sodium salt or a trifluoroacetic acid salt. 17. A compound which is 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol- -yl)-2-(((R)-1 ,2- dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3- yi)methyi)-1~((1 -ethyl~8,7-dihydroxy-4~oxo~1 ,4-dihydroquinolin-3-yl)methyl)pyrrolidin-1-

!um:

a pharmaceutically acceptable salt thereof.

18. A compound which is 1-(((6R,7R)-7-((Z)-2-(2-aminothiazol- -yl)-2-(((R)-1 ,2- dicarboxyethoxy)imino)acetamido)-2-carboxy-8-oxo-5-thia-1-azabicycio[4.2.0]oct-2-en-3- yi)methyi)-1~((1 -ethyl~8,7~dihydroxy-4~oxo~1 ,4-dihydroquino!in-3-yl)methyl)pyrrolidin-1 -ium disodium salt.

19. A compound which is 1-(((6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yi)-2-(((2- carboxypropan-2-yi)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct- PU85023

2-en-yl)met yi)-1-((1 -ethyl-6 -dihydroxy-4-oxo-1 ,4-dihydrocinnolin-3-yl)methyi)pyrrolidin'

a pharmaceutically acceptable salt thereof.

20. A compound which is 1-(((6R,7R)-7-((Z)-2-(2-aminothiazoi-4-yi)-2-(((2- carboxypropan-2-yi)oxy)imino) acetamido)-2-carboxy-8-oxo-5-thia-1-azabicyclo[4.2.0]oct- 2-en-3-yi)methyl)-1 -(2-(1 -ethyl-6,7-dihydroxy-4-oxo-1 ,4-dihydrocinnoline-3- carboxamido)ethyl)pyrroiidin-1-ium;

a pharmaceutically acceptable salt thereof.

21 , A pharmaceutical composition comprising a compound according to any one of claims 1 to 20 and at least one or more pharmaceutically acceptable excipients,

22, A method of treating a bacterial Infection comprising administering a

therapeutically effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof or a pharmaceutical composition according to claim 21 to a human in need thereof.

23, The method of treating a bacterial infection according to claim 22, wherein the bacteria! infection is caused by Gram negative bacteria. PU85023

24. The method of treating a bacterial infection according to claim 23, wherein the Gram negative bacteria selected from Gram negative bacteria of enterobacteria, Gram negative bacteria colonized in respiratory, Gram negative bacteria of glucose non fermentation or β-iactam drug resistant Gram negative bacteria.

25. The method of treating a bacterial infection according to claim 24, wherein: the Gram negative bacteria of enterobacteria selected from E. coii, Klebsiella,

Serratia, Enterobacter, Cltrobacter, organeiia, Providencia or Proteus;

the Gram negative bacteria colonized in respiratory system selected from Haemophilus or Moraxe!la;

the Gram negative bacteria of glucose non fermentation selected from

26. The method according to claim 22, wherein the bacteria! infection is an airway Infection, urinary system infection, resipiratory system infection, sepsis Infection, nephritis, cholecystitis, oral cavity infection, endocarditis, pneumonia, bone marrow membrane myelitis, otitis media, enteritis, empyema, wound infection or an opportunistic infection .

27. A method for treating a gram-negative bacterial infection comprising administering a therapeuticaiiy effective amount of a compound according to any one of claims 1 to 20 or a pharmaceutically acceptable salt thereof to a human in need thereof.

28. The method of treating a gram-negative bacterial infection according to claim 27, wherein the bacterial infection is caused by Gram negative bacteria. 29. The method of treating a gram-negative bacterial infection according to claim 28, wherein the Gram negative bacteria selected from Gram negative bacteria of enterobacteria, Gram negative bacteria colonized in respiratory, Gram negative bacteria of glucose non fermentation or β-iactam drug resistant Gram negative bacteria. 30. The method of treating a gram-negative bacterial infection according to claim 29, wherein: PU85023 the Gram negative bacteria of enterobacteria selected from E. colL Klebsiella, Serratia, Enterobacter, Citrobacter, organe!!a, Providencia or Proteus;

the Gram negative bacteria of glucose non fermentation selected from

Pseudomonas aeruginosa, Pseudomonas other than P. aeruginosa, Stenotrophomonas or Burkholderia, Aclnetobacter; and

the beta-lactam drug resistant Gram negative bacteria is selected from ESBL producing bacteria.

31 . A method for inhibiting activity of UDP-3-0— (R-3-hydroxydecanoyl)-N- acetylg!ucosamine deacetylase (LpxC) comprising administering a therapeutically effective amount of a compound of Formula (H) according to anyone of claims 1 to 20 or a pharmaceutically acceptable salt thereof to a human in need thereof.

32. A method for treating antimicrobial activity against Gram positive bacteria comprising administering a therapeutically effective amount of a compound according to anyone of claims 1 to 20 or a pharmaceutically acceptable salt thereof to a human in need thereof.

33. The method for treating antimicrobial activity against Gram positive bacteria according to claim 32, wherein the Gram positive bacteria is selected from methicillin- resistant Staphylococcus aureus ( RSA) or penicillin-resistant Streptococcus pneumoniae (PRSP).

34. A method for treating methicl!lin-resistant Staphylococcus aureus (MRSA), comprising administering a therapeutically effective amount of a compound according to anyone of claims 1 to 20 or a pharmaceutically acceptable salt thereof to a human in need thereof.

35. A method for treating penicillin-resistant Streptococcus pneumoniae (PRSP), comprising administering a therapeutically effective amount of a compound according to anyone of claims 1 to 20 or a pharmaceutically acceptable salt thereof to a human in need thereof.