WO2022047790A1

WO2022047790A1 - Amidine substituted bicyclic compounds, their preparation, their use as antibacterial agents and beta-lactamase inhibitors

Info

Publication number: WO2022047790A1
Application number: PCT/CN2020/113787
Authority: WO
Inventors: Zhixiang Yang; Haikang YANG; Jinbo JI; Jian Sun; Yuanyu GAO; Dong TANG; Jingwen JI; Lijuan ZHAI; Yangxiu MU; Zafar Iqbal; Lili He; Yuanbai LIU; Rui JIANG; Koko MYO; Zawmin THU; Xueqin Ma; Jianqiang Yu
Original assignee: Ningxia Academy of Agriculture and Forestry Sciences
Current assignee: Ningxia Academy of Agriculture and Forestry Sciences
Priority date: 2020-09-07
Filing date: 2020-09-07
Publication date: 2022-03-10
Anticipated expiration: 2023-03-07

Abstract

It is directed to amidine substituted bicyclic compounds which are β-lactamase inhibitors, for the treatment of bacterial infections, including infection caused by drug resistant organisms, including multi-drug resistant organisms. It includes compounds according to formula (I) : or pharmaceutically acceptable salts thereof, wherein the values of M and Y are as defined herein.

Description

Amidine substituted bicyclic compounds, their preparation, their use as antibacterial agents and beta-lactamase inhibitors

Field of the invention

The present invention relates to amidine substituted bicyclic compounds, their preparation and their use as antibacterial agents either alone or in combination with an antibiotic (or plural antibiotics) for the treatment of infections caused by β-lactamase-producing pathogenic bacteria. The compounds of the present invention are β-lactamase inhibiting or non-β-lactamase inhibiting (i.e., some of the present invention by themselves would directly inhibit β-lactamase enzymatic function, and others of compounds of the present invention by themselves would not inhibit some β-lactamase enzymatic function though they provide synergy and increased potency of activity in combination with antibiotics, e.g., β-lactam antibiotics or non-β-lactam antibiotics) . More particularly, The present invention is concerned with methods for overcoming antibiotic resistance caused by β-lactamase producing bacteria, the method of preparation of the amidine substituted bicyclic compounds, pharmaceutical compositions containing the amidine substituted bicyclic compounds, methods of treatment, uses of the compounds, and other subject matter.

Background of the invention

Microbial drug resistance to β-lactam antibiotics, especially in Gram-negative bacterial, is most commonly mediated by β-lactamases, β-lactamases are enzymes that catalyze the hydrolysis of the β-lactam ring, which inactivates the antibacterial activity of the β-lactam antibiotic and allow the bacterial to become resistant. Inhibition of the β-lactamase with a β-lactamases inhibitor slows or prevents degradation of the β-lactam antibiotic and restores β-lactam antibiotic susceptibility to β-lactamase producing bacteria. Many of these β-lactamases are not effectively inhibited by β-lactamase inhibitors currently on the market rendering the β-lactam antibiotics ineffective in treating bacteria that producing these β-lactamases. There is an urgent need for novel β-lactamase inhibitors that inhibit β-lactamases that are not effectively inhibited by the current clinical β-lactamases (e.g. KPC, class C and class D β-lactamases) and that could be used in combination with β-lactam antibiotics to treat infection caused by β-lactam resistant bacteria.

Recently, certain diazabicyclic compounds have been disclosed in WO 2009/091856 which is hereby incorporated by reference in its entirety. In addition, a number of diazabicyclic heterocycles have been disclosed in the following patents or applications as β-lactamase inhibitors: US 2003/0199541 A1, US 2004/0157826 A1, US 2004/0097490 A1, US 2005/0020572 A1, US 2006/7112592 B2, US 2006/0189652 A1, US 2008/7439253 B2, US 2009/0018329 A1, EP 1307457 B1, EP 1537117 B1, WO 2002/100860 A2, WO 2002/10172 A1, WO 2003/063864 A2, WO 2004/052891 A1, WO 2004/022563 A1, WO 2008/142285 A1, WO 2009/090320 A1, US 2010/0092443 A1, WO 2010/126820 A2, WO 2013/038330 A1, US 2015/0031666 A1, US 2015/0239840 A1, US 2016/0297817 A1, US2016/0002235 A1, WO 2017037607 A1, WO 2018053057 A2, WO 2018053215 A1.

The compounds of the present invention are new and the structural features are significantly distinct from the compounds described in the patent references cited above.

Summary of the invention

In one aspect, the present invention relates to new diazabicyclic compounds (some of which have potent broad-spectrum β-lactamase inhibitory activity and others do not have such activity) that when used in combination with a β-lactam antibiotic or with other non β-lactam antibiotic enhance the activity of the antibiotic against class A, class B, class C, and class D enzyme producing organisms and thereby enhance the antibacterial properties. The inventive compounds are therefore useful in the treatment of bacterial infections in humans or animals either alone or in combination with β-lactam antibiotics.

In accordance with the present invention, there are provided (A) new compounds of general formula (I) , (B) pharmaceutically acceptable salts of the compounds of formula (I) , and (C) pharmaceutically acceptable solvates of the compounds of formula (I) and of their salts, and (D) deuterated compounds of compounds of (A) , (B) and (C) , (namely, (i) compounds of formula (I) modified in that they have been deuterated, (ii) pharmaceutically acceptable salts of the compounds of formula (I) modified in that they have been deuterated, (iii) pharmaceutically acceptable solvates of the compounds of formula (I) and of their salts modified in that they have been deuterated) :

wherein:

M is hydrogen or a pharmaceutically acceptable salt forming cation,

a “pharmaceutically acceptable salt” refers to a salt of a compound, which salt possesses the desired pharmacological activity of the parent compound,

reference to specified compounds “modified in that they have been deuterated” refers to compounds prepared by modifying the specified compounds so that one or more hydrogen atoms in the compound have been replaced with or converted to deuterium,

Y = H or R,

Wherein, R is optionally substituted with one or two substituents independently selected from the following:

Lower alkyl, amino, substituted amino, alkoxy, hydroxyalkyl, halogen, hydroxy, carboxy, alkoxycarbonyl, haloalkyl, trifluoromethyl, trifluoromethyloxy, alkylamine, substituted alkylamine, carboxamide, thiocarboxamide, sulfonic acid, sulphate, acylamino, sulfonylamino, substituted or unsubstituted sulfonamide, substituted or unsubstituted urea, substituted or unsubstituted thiourea, oxyimino, hydroxamic acid, acyl, trifluoromethyl carbonyl, cyano, amidino, guanidino, aryloxy, heterocyclylalkyloxy, and heteroaryloxy.

The compounds of the present invention are new and the structural features are significantly distinct from the compounds described in the prior art.

In the formula (I) , when Y = R, R is a radical selected from any of the following groups:

(1) C _1-6 straight or branched chain alkyl which is optionally substituted. Non-limiting examples of such compounds are:

(2) C _3-7 cycloalkyl which is optionally substituted. Non-limiting examples of such compounds are:

(3) C _4-7 saturated heterocycles containing at least one heteroatom selected from O, N and S wherein the said heterocycle is optionally substituted. Furthermore the ring S is optionally oxidized to S (O) or S (O) ₂ and the free ring N atom may optionally take a substituent. Non-limiting examples of such compounds are:

(4) Heterocyclyl (C _1-6) alkyl wherein the said heterocycle has the same definition as defined in (3) . Furthermore, the said heterocycle is optionally substituted. Non-limiting examples of such compounds are:

(5) C _5-7 membered heteroarylalkyl which is optionally substituted. Non-limiting examples of such compounds are:

Examples of the groups for forming a pharmaceutically acceptable salt represented by M in the formula (I) include: inorganic base salts, ammonium salts, organic base salts, basic amino acid salts, inorganic acid addition salts, and organic acid addition salts. Inorganic bases that can form the inorganic base salts include alkali metals such as sodium, potassium, and lithium and alkaline earth metals such as calcium and magnesium. Organic bases that can form the organic base salts include n-propylamine, n-butylamine, cyclohexylamine, benzylamine, octylamine, ethanolamine, diethanolamine, diethylamine, triethylamine, dicyclohexylamine, procaine, choline, N-methylglucamine, morpholine, pyrrolidine, piperidine, N-ethylpiperidine and N-methylmorpholine.

Basic amino acids that can form the basic amino acid salts include lysine, arginine, ornithine and histidine. As will be appreciated by one skilled in the art, the compounds of formula (I) containing a basic nitrogen atom are capable of forming acid addition salts. Such salts with pharmaceutically acceptable acids are included in the invention. Examples of such acids are hydrochloric, hydrobromic, phosphoric, sulphuric, citric, oxalic, maleic, fumaric, glycolic, mandelic, tartaric, aspartic, succinic, malic, formic, acetic, p-toluenesulfonic, trifluoroacetic, methanesulfonic, ethanesulfonic, trifluoromethanesulfonic, benzenesulfonic and the like.

Moreover, some compounds of formula (I) when they contain a basic group such as NH, NH ₂ or pyridine and the like may form an inner, zwitterionic salt with OSO ₃H; such inner salts are also included in this invention.

Another aspect of the present invention is to include all possible isomers of formula (I) . As used herein, the term ‘isomers’ refers to different compounds that have the same molecular formula but differ in arrangement and configuration of the atoms, such as geometrical isomers and optical isomers. For a given compound of the present invention, it is understood that a substituent may be attached at a chiral center of a carbon atom. Therefore the invention includes enantiomers, diastereomers or racemates of the compound. By definition ‘enantiomers’ are a pair of stereoisomers that are non-superimposable mirror images of each other, and 1: 1 mixture of a pair of enantiomers is a racemic mixture. By definition, ‘diastereoisomers’ are stereoisomers that have at least two asymmetric carbon atoms but which are not mirror-images of each other. When a compound of formula (I) is a pure enantiomer the stereochemistry at each chiral carbon may be specified by either R or S.

Compounds may also exist in several tautomeric forms including the enol form, the keto form, and mixtures of any of the foregoing. Accordingly, the chemical structures depicted herein encompass all possible tautomeric forms of the illustrated compounds.

A variety of protecting groups conventionally used in the β-lactam field to protect a reactive functional group present in the molecule of formula (I) can be used. ‘Protecting group’ refers to a group of atoms that when attached to a reactive functional group in a molecule masks, reduces or prevents reactivity of the functional group. Examples of protecting groups can be found in “Protective Groups in Organic Synthesis” , (Theodora W. Greene and Peter G.M. Wuts, John Wiley &Sons. Inc., 3 ^rd, 1999) . Representative amino protecting groups include, but are not limited to formyl, acetyl, trifluoroacetyl, benzyl, benzyloxycarbonyl (CBZ) , tert-butoxycarbonyl (Boc) , trimethylsilyl (TMS) , 9-fluorenylmethyloxycarbonyl (FMOC) , nitro-veratryoxycarbonyl (NVOC) , and the like. Examples of hydroxy protecting groups include, but are not limited to, those where the hydroxyl group is either acylated or alkylated such as benzyl, and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers, trialkylsilyl ethers, and allyl ethers.

The term ‘optionally substituted’ refers to unsubstituted or substituted with one or two of the following substituents each of which is independently selected from:

Lower alkyl including from one to six carbon atoms in any arrangement, e.g., methyl, ethyl, i-propyl or t-butyl,

Amino,

Substituted amino such as –NHCH ₃, -N (CH ₃) ₂ , -NHCH ₂CH ₃, -NHPr ⁱ, -NHBu ^t,

Alkoxy such as –OCH ₃, -OC ₂H ₅ , -OPr ⁱ (i.e., isopropyloxy) , -OBu ^t (i.e., isobtutyloxy) ,

Hydroxyalkyl such as –CH ₂OH, -CH ₂CH ₂OH,

Halogen such as F, Cl, Br , I,

Hydroxy,

Carboxy,

Alkoxycarbonyl such as –COOCH ₃, -COOC ₂H ₅, -COOPr ⁱ, and -COOBu ^t,

Haloalkyl such as –CH ₂Cl, -CH ₂F,

Trifluoromethyl,

Trifluoromethyloxy,

Alkylamine such as –CH ₂NH ₂, –CH ₂CH ₂NH ₂,

Substituted alkylamine such as –CH ₂NHCH ₃, –CH ₂N (CH ₃) ₂, –CH ₂CH ₂NHCH ₃, –CH ₂CH ₂N (CH ₃) ₂,

Carboxamide,

Thiocarboxamide,

Sulfonic acid,

Sulfate,

Acylamino,

Sulfonylamino,

Sulfonamide,

Substituted sulfonamide such as –SO ₂NHCH ₃, –SO ₂NHCH ₂CH ₃, –SO ₂NHPr ⁱ, –SO ₂NHBu ^t,

Urea (-NHCONH ₂) which may be optionally substituted,

Thiourea (-NHCSNH ₂) , optionally substituted,

Sulfonylurea (-NHSO ₂NH ₂) , optionally substituted,

Oxo (=O) when oxygen is bonded through double bond to a carbon atom,

Oxyimino (=N-O-A) where the nitrogen is bonded through double bond to a carbon atom which is attached to the rest of the molecule and A can be hydrogen, or optionally substituted straight or branched lower alkyl, cycloalkyl, aryl, heteroaryl and heterocyclyl,

Hydroxamic acid (-CONHOH) ,

Acyl (-COCH ₃) ,

Trifluoromethyl carbonyl (-COCF ₃) ,

Cyano (-CN) ,

Amidino -C (=NH) NH ₂ which may be optionally substituted,

Guanidino -NHC (=NH) NH ₂ which may be optionally substituted,

Aryloxy,

Heterocyclyl,

Heteroaryl,

Heterocyclyloxy,

Heteroaryloxy,

Heterocyclyalkyloxy,

Trialkylammonium,

The substituent mentioned above could be substituted at the carbon atom or at the free N-atom of the molecule as appropriate.

Among the compounds of formula (I) , a particular subject of the invention is those in which M is hydrogen or a pharmaceutically acceptable salt forming cation.

A group of preferred examples of formula (I) , when Y = H or R, are from the following Table 1.

* = point of attachment with N

Table 1. List of compounds

It is also an object of this invention to provide a combination of a compound of general formula (I) having antibacterial activity with another existing antibacterial agent, thus causing synergistic effect and the use of the same as drugs for the treatment of bacterial infections.

It is another object of the invention to provide methods for preparing the compounds of the invention of formula (I) .

It is a further object of the invention to provide pharmaceutical compositions comprising a compound of formula (I) of this invention as an active ingredient in combination with an antibiotic (e.g., a β-lactam antibiotic or some other non β-lactam antibiotic) and a suitable amount of pharmaceutically acceptable carrier or diluent, so as to provide a form for proper administration to a patient. These compositions can be administered by parenteral, in particular intramascular route, oral, sublingual, rectal, aerosol or by local route in a topical application on the skin and the mucous membranes. Suitable pharmaceutical vehicles include excipients such as starch, glucose, lactose, sucrose, gelatin, gum arabic, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, and the like. Other examples of suitable pharmaceutical vehicles have been described in the art (Remington’s Science and Practice of Pharmacy, 21 ^st Edition, 2006) . Compositions of the present disclosure, if desired, can also contain minor amounts of wetting, dispersing or emulsifying agents, or pH buffering agents, and preservatives. In addition, auxiliary, stabilizing, thickening, lubricating, and coloring agents can be included. Pharmaceutical compositions can be formulated in a conventional manner. Proper formulation is dependent upon the route of administration chosen. The present pharmaceutical compositions can take the form of injectable preparations, suspensions, emulsions, sugar-coated tablets, pellets, gelatin-capsules, capsules containing liquids, powders, granules, sustained-release formulations, suppositories, aerosols, sprays, ointments, creams or any other form suitable for use.

In another aspect, the present invention also provides for the use, in the manufacture of a medicament, of a compound within formula (I) above as an active ingredient in an antibacterial composition in admixture with a carrier.

In another aspect, the present invention also provides for the use, in the manufacture of a medicament, of a compound within formula (I) above as an active ingredient.

In another aspect, the present invention also provides for the use, in the manufacture of a medicament, of a compound within formula (I) above as an active ingredient, along with one or more β-lactam antibiotics (e.g., a β-lactam antibiotic or some other non β-lactam antibiotic) , in an antibacterial composition in admixture with a carrier.

In another aspect, the present invention also provides for the use, in the manufacture of a medicament, of a compound within formula (I) above as an active ingredient, along with one or more β-lactam antibiotics (e.g., a β-lactam antibiotic or some other non β-lactam antibiotic) .

The parenteral administration which includes intramuscular, intraperitonial, subcutaneous and intravenous use, sterile solutions of the active ingredient are usually prepared and the pH of the solutions are suitably adjusted and buffered. For intravenous use, the total concentration of solutes should be controlled to render the preparation isotonic. Suitable solvents include saline solution (e.g., 0.9%NaCl solution) and apyrogenic sterile water. Pharmaceutical compositions for oral delivery can be, for example, in the form of tablets, lozenges, aqueous or oily suspensions, granules, powders, emulsions, capsules, syrups, or elixirs. Orally administered compositions can contain one or more optional agents, for example, sweetening agents such as fructose, aspartame, or saccharin, flavoring agents such as peppermint, oil of wintergreen, cherry, coloring agents, and preserving agents to provide a pharmaceutically palatable preparation. Moreover, when in tablet form, the compositions can be coated to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over an extended period of time. Oral compositions can include standard vehicles such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, cellulose, magnesium carbonate, and the like. For oral liquid preparations, for example, suspensions, elixirs, and solutions, suitable carriers, excipients, or diluents include water, saline, alkyleneglycols (e.g. propylene glycol) , polyalkylene glycols (e.g., polyethylene glycol) , oils, alcohols, slightly acidic buffers ranging from about pH 4 to about pH 6 (e.g., acetate, citrate, ascorbate ranging from about 5 mM to about 50 mM) , and the like. Additionally, flavoring agents, preservatives, coloring agents, bile salts, acylcarnitines, and the like can be added.

For topical formulations of compounds of the present invention, creams, gels, ointments or viscous lotions can be used as appropriate delivery forms. Topical delivery systems also include transdermal patches containing at least one compound of formula (I) to be administered.

Delivery through the skin can be achieved by diffusion or by more active energy sources such as iontophoresis or electrotransport. Formulations of a compound of the present invention, for topical use, such as in creams, ointments, and gels, can include an oleaginous or water soluble ointment base, for example, topical compositions can include vegetable oils, animal fats, and in certain embodiments, semisolid hydrocarbons obtained from petroleum. Topical compositions can further include white ointment, yellow ointment, cetyl esters wax, oleic acid, olive oil, paraffin, petrolatum, white petrolatum, spermaceti, starch glycerite, white wax, yellow wax, lanolin, and glyceryl monostearate. Various water-soluble ointment bases can also be used, including glycol ethers and derivatives, polyethylene glycols, polyoxyl 40 stearate, and polysorbates.

In a pharmaceutical composition containing a compound of this invention, the weight ratio of active ingredient to carrier will normally be in the range of 1: 30 to 30: 1. The administered daily dose varies according to the illness treated, and the administration route. However in most instances, an effective dose (e.g., in some instances, β-lactamase inhibiting dose) of a compound of formula (I) or a pharmaceutically acceptable salt thereof will be a daily dose in the range from about 1 to about 500 mg per kilogram of body weight orally, and from about 1 to about 500 mg per kilogram of body weight parenterally. The weight ratio of the compound of present invention to the antibiotic (if it is being administered with an antibiotic) will normally be in the range from 1: 30 to 30: 1.

In some aspects of the present invention, an additional object is to provide an improved method for the treatment of bacterial infections caused by β-lactamase producing bacteria in a patient in need of such treatment comprising administering to the patient a therapeutically effective amount of at least one compound chosen from formula (I) or a pharmaceutically acceptable salt thereof in combination with a known β-lactam antibiotic. In such an aspect of the present invention, the compounds increase the antibacterial effectiveness of β-lactamase susceptible β-lactam antibiotics, that is, they increase the effectiveness of the antibiotic against infections caused by β-lactamase producing microorganisms in mammalian subjects, particularly in human. In these aspects of the present invention, this makes the compounds of formula (I) and pharmaceutically acceptable salts thereof, valuable for co-administration with β-lactam antibiotics. In the treatment of a bacterial infection in such an aspect of the present invention, said compounds of formula (I) or a pharmaceutically salt thereof can be mixed with the β-lactam antibiotic, and the two agents thereby administered simultaneously. When co-administered with a β-lactam antibiotic in such an aspect of the present invention, the combination of the compound of the invention and the antibiotic can provide a synergistic effect. The term ‘synergystic effect’ refers to the effect produced when two or more agents are co-administered is greater than the effect produced when the agents are administered individually. Alternatively, the compound of formula (I) or a salt thereof can be administered as a separate agent during a course of treatment with the antibiotic.

‘Therapeutically effective amount’ refers to the amount of a compound that, when administered to a subject for treating a disease, or at least one of the clinical symptoms of a disease, is sufficient to affect such treatment of the disease, disorder, or symptom. The therapeutically effective amount can vary depending, for example, on the compound, the disease, disorder, and/or symptoms of the disease, severity of the disease, disorder, and/or symptoms of the disease, the age, weight, and/or health of the patient to be treated, and the judgement of the prescribing physician.

The term ‘β-lactam antibiotic’ refers to a compound with antibiotic property that contains a β-lactam functionality. Examples of β-lactam antibiotics which can be used in combination with the compounds of the present invention represented by formula (I) are commonly marketed penicillins, cephalosporins, penems, carbapenems and monobactams.

Examples of β-lactam antibiotics which can be used in combination with the compounds of the present invention represented by formula (I) are commonly used penicillins, such as amoxicillin, ampicillin, azlocillin, mezlocillin, apalcillin, hetacillin, bacampicillin, carbenicillin, sulbenicillin, ticarcillin, piperacillin, methicillin, ciclacillin, talampicillin, oxacillin, cloxacillin, dicloxacillin and commonly used cephalosporins such as cephalothin, cephaloridine, cefaclor, cefadroxil, cefamandole, cefazolin, cephalexin, cephradine, cephapirin, cefuroxime, cefoxitin, cephacetrile, cefotiam, cefotaxime, cefatriazine, cefsulodin, cefoperazone, ceftizoxime, cefmenoxime, cefmetazole, cephaloglycin, cefonicid, cefodizime, cefpirome, cefepime, ceftazidime, cefpiramide, ceftriaxone, cefbuperazone, cefprozil, cefixime, ceftobiprole, ceftaroline, cefalonium, cefminox, ceforanide, cefuzonam, cefoxitin, cefotetan, loracarbef, cefdinir, cefditoren, cefetamet, cefcapene, cefdaloxime, ceftibuten, cefroxadine and latamoxef (moxalactam) . From the carbapenem class of β-lactam antibiotics such as imipenem, meropenem, panipenem, biapenem, doripenem, ertapenem and the like could be used. From monobactam class of β-lactam antibiotics such as aztreonam, carumonam, tigemonam, and the like could be used as the combination partner of antibiotic.

Examples of antibiotics (which are not β-lactam antibiotics) which can be used in combination with the compounds of the present invention (i.e., compounds of formula (I) above, salts, thereof, solvates of such compounds and salts, and deuterated compounds of any such compounds) include aminoglycosides, quinolones, tetracyclines, glycylcyclines, glycopeptides, lipopeptides, macrolides, ketoliddes, lincosamides, streptogramin, oxazolidinones, polymyxins, and other compounds known to have antibacterial properties.

‘Pharmaceutically acceptable solvate’ refers to a molecular complex of a compound with one or more solvent molecules in a stoichiometric or non-stoichiometric amount. Such solvent molecules are those commonly used in the pharmaceutical art, which are known to be innocuous to recipient, e.g., water, ethanol, and the like. A molecular complex of a compound or moiety of a compound and a solvent can be stabilized by non-covalent intra-molecular forces such as, for example, electrostatic forces, Van der Waals forces or hydrogen bonds. The term hydrate refers to a complex where the one or more solvent molecules are water.

Among the compounds of formula (I) , a particular subject of the invention is the compounds with the following names. The following examples illustrate the invention, and are not intended to be limiting of its scope. To the contrary, the claims are intended to cover alternatives, modifications, and equivalents.

The non-limiting examples of the compounds of the present invention are:

(2S, 5R) -2-Carbamimidoyl-7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Methylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Ethylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Isopropylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N-propylcarbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Isobutylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Neopentylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (Cyclopentylmethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (Cyclohexylmethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Acetamidoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Aminoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Hydroxyethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-oxo-2- (N- (2-Ureidoethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(Imino ( (2S, 5R) -7-oxo-6- (sulfooxy) -1, 6-diazabicyclo [3.2.1] octan-2-yl) methyl) glycine,

(2S, 5R) -2- (N- (2-Methoxyethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (piperazin-1-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (tetrahydropyrimidin-1 (2H) -yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (piperidin-4-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (4-Methylpiperazin-1-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (3-Acetyltetrahydropyrimidin-1 (2H) -yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (1-Methylpiperidin-4-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Aminopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Amino-2-oxoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (piperidin-4-yloxy) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (pyrrolidin-3-yloxy) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (Azetidin-3-yloxy) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (piperidin-1-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (1-Acetylpiperidin-4-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (pyrrolidin-3-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (Azetidin-3-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (tetrahydro-2H-pyran-4-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (2- (pyrrolidin-1-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (1, 1-Dioxidotetrahydro-2H-thiopyran-4-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Guanidinoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (Guanidinooxy) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (1-Acetylpyrrolidin-3-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-cyclopropylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Cyclobutylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Cyclopentylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3-Aminocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3- (Dimethylamino) cyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3- (Methylamino) cyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3-Acetamidocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Aminocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (Dimethylamino) cyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Acetamidocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Acetamidocyclopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2- (Methylamino) cyclopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (4-Aminocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (4- (Dimethylamino) cyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N-Cycloheptylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (4-Acetamidocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3-Aminocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3- (Dimethylamino) cyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3- (Methylamino) cyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (3-Acetamidocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (2-Aminocyclopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (piperidin-4-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (1-Methylpiperidin-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (1-Acetylpiperidin-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

Ethyl 4- ( (2S, 5R) -7-oxo-6- (sulfooxy) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) piperidine-1-carboxylate,

(2S, 5R) -7-Oxo-2- (N- ( (R) -piperidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (R) -1-Methylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (R) -1-Acetylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- ( (S) -piperidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (S) -1-Methylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (S) -1-Acetylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- ( (S) -pyrrolidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (S) -1-Ethylpyrrolidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- ( (R) -pyrrolidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (R) -1-Formylpyrrolidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (R) -1-Acetylpyrrolidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (Azetidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (1-sulfamoylazetidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (1- ( (l2-Azanyl) (imino) methyl) azetidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (Azepan-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (tetrahydro-2H-thiopyran-4-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (tetrahydro-2H-pyran-4-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (1, 1-Dioxidotetrahydro-2H-thiopyran-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- (1, 4-Oxazepan-6-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (pyrrolidin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (piperidin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (piperidin-4-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (piperidin-2-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (1, 3-Oxazinan-2-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (piperazin-2-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (1, 3-Thiazinan-2-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- ( (tetrahydrofuran-2-yl) methyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-oxo-2- (N- ( (tetrahydro-2H-pyran-4-yl) methyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (pyridin-4-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (pyridin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (pyrimidin-5-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -7-Oxo-2- (N- (pyridin-2-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (1H-Imidazol-2-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

(2S, 5R) -2- (N- ( (1H-Imidazol-1-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate,

The present invention also relates to methods for the preparation of compounds of formula (I) . The compounds of the present invention of formula (I) can be readily prepared by the following reaction Scheme 1 and examples using readily available starting materials, reagents and conventional synthesis procedures known to those of ordinary skill in this art.

SCHEME 1

The bicyclic intermediate amide (II) may be prepared following the literature (Org. Process Res. Dev. 2016, 20, 1799-1805) .

Compounds of the general of formula (I) can be prepared by converting bicyclic amide (II) to bicyclic nitrile (III) in presence of a suitable reagents. The suitable reagents used for carrying out this step include, but are not limited to trifluoroacetic anhydride (TFAA) and triethylamine (TEA) or diisopropylethylamine (DIPEA) , phosphoryl chloride (POCl ₃) and TEA, and the like. The organic solvents useful in the reaction are not particularly limited and include any of those which do not adversely affect the reaction. Typical solvents include dichlomethane, chloroform, tetrahydrofuran and the like. The reaction is normally carried out at a temperature of from about 0 ℃ to 40 ℃, and preferably at room temperature under nitrogen. After completion of the reaction the desired product can be easily separated by conventional methods such as column chromatography, crystallization or similar methods.

The unsubstituted amidine (IV) can be prepared by reacting NH ₄Cl with he nitrile (III) , and the substituted amidine (IV) can be prepared by substituting an appropriately substituted amine (NH ₂-Y) to the nitrile (III) in presence of a suitable reagent. The suitable reagents useful for carrying out this step include, but are not limited to trimethylaluminum, or triethylaluminum, or trifluoromethanesulfonate, or Lanthanum (III) and the like. The organic solvents useful in the reaction are not particularly limited and include any of those which do not adversely affect the reaction. Typical solvents include dichloromethane, chloroform, toluene, dimethylformamide, tetrahydrofuran, dimethylsulfoxide, and the like. The reaction is normally carried out at a temperature of from about 0 ℃ to about 100 ℃. After completion of the reaction the desired product can be easily separated by conventional methods such as column chromatography, crystallization or similar methods.

The intermediate (V) may be prepared by putting a protection groups (p) on the substituted amidine (IV) . The protecting group can be Boc (tert-butoxycarbonyl) , Cbz (benzyloxycarbonyl) , trifluoroacetyl and the like.

In the following step, the intermediate (V) could be converted to compound (VI) under an atmosphere of hydrogen or hydrogen mixed with an inert diluent such as nitrogen or argon in the presence of a hydrogenation catalyst. The catalysts used in this hydrogenation reaction are the type of agents known in the art for this kind of deprotection and typical examples are the noble metals, such as nickel, palladium, platinum and rhodium. Examples of the catalysts are platinum, platinum oxide, palladium, palladium oxide and the like. The catalyst is usually present in the amount from about 1 to about 50 weight percent and preferably from about 5 to about 15 weight percent based on the compound of V. It is often convenient to suspend the catalyst on an inert support. A particularly convenient catalyst is palladium suspended on an inert support such as carbon, e.g. 5%or 10%by weight palladium on carbon. This reaction may be conveniently effected at ambient temperature from 15 psi to 60 psi until reaction is complete (2 to 72 hours) . Suitable solvents for this reaction are those which substantially dissolve the starting material of the formula (V) , after reaction, the suitable solvents are sufficiently volatile to be removed by evaporation and do not themselves suffer hydrogenation. Examples of such solvents include methanol, ethanol, dioxane, ethyl acetate, tetrahydrofuran or a mixture of these solvents. Upon completion, the hydroxy intermediate (VI) can be purified by silica gel column chromatography or in many cases can be directly carried out to the next step without further purification.

Sulfation of the intermediate (VI) can be achieved using a sulfating reagent (e.g., pyridine-SO ₃ complex, NMe ₃-SO ₃ complex, DMF-SO ₃ complex and ClSO ₃H) in an appropriate base (e.g., pyridine, triethylamine or 2-picoline) as described in the literature (WO2017155765A1, Org. Process Res. Dev. 2016, 20, 1799-1805) . Thus, pyridine-SO ₃ complex can be added to a solution of the intermediate (VI) in a solvent in excess amount, if desired, to force the reaction to completion. The organic solvents useful for this transformation are not particularly limited and include those which do not adversely affect the reaction. Typical solvents include, but not limited to, pyridine, tetrahydrofuran, isopropyl alcohol and water, dimethyl formamide, dimethylacetamide, acetonitrile, dichloromethane, and the like. The transformation can be carried out at from 0 ℃ to 40 ℃, and more preferably at room temperature. The product (VII) can be isolated by standard procedure that is by filtering the reaction mixture, concentrating the filtrate, which was directly used for next step without further purification. Sometime the filtrate concentrated may need to purified silica gel column chromatography to give desired compound (VII) .

Finally, the compound of formula (I) can be achieved by treating the intermediate (VII) with an acid to remove protecting group when the intermediate (VII) containing protection group, such as Boc. and the like, or the compound of formula (I) can also be achieved by treating the intermediate (VII) with an base to remove protecting group when the intermediate (VII) containing protection group, such as trifluoacetyl and the like. Suitable acids include trifluoroacetic acid, methanesulfonic acid, trifluoromethane sulfonic acid, formic acid and the like. Suitable bases include K ₂CO ₃, Na ₂CO ₃, NaOH, KOH, LiOH and the like The treatment is suitably conducted at a temperature in a range from about -10 ℃ to about 100 ℃ and is typically conducted at a temperature in a range of from about 0 ℃ to about 35 ℃. Suitable purification methods for the final compound of formula (I) are normal silica gel chromatograph, perp. HPLC, HP20 chromatograph, inon exchange resin and the like.

Detailed description of the invention

Abbreviations

In the experiments the following abbreviations have been used:

δ: chemical shift in parts per million (ppm) by frequency

br s: broad single in NMR

d: doublet in NMR

dd: doublet of doublet in NMR

t: triplet in NMR

q: quartet in NMR

m: multiplet in NMR

J: coupling constant in NMR

Hz: hertz

MHz: megahertz

NMR: nuclear magnetic resonance

ES ^-: negative ion mode in electrospray ionization mass spectrometry

ES ⁺: positive ion mode in electrospray ionization mass spectrometry

MS: mass spectrum

HPLC: high performance liquid chromatography

g: gram (s)

mg: milligram (s)

mmol: millimole (s)

mol: mole (s)

L: liter (s)

mL: milliliter (s)

M: molarity

h: hour (s)

min: minute (s)

Pd/C: palladium on carbon

DCM: dichloromethane

DMF: dimethylformamide

DMSO: dimethyl sulfoxide

EtOAc: ethyl acetate

TFA: trifluoroacetic acid

THF: tetrahydrofuran

TLC: thin layer chromatography

TMS: tetramethylsilane

CDCl ₃: deuterated chloroform

CD ₃OD: deuterated methanol

D ₂O: deuterium oxide

pH: the negative logarithm of the hydrogen ion concentration

Boc: N-tert-butoxycarbonyl

Bn: benzyl

Cbz: benzyloxycarbonyl

HPLC: high-performance liquid chromatography

δ: chemical shift in parts per million (ppm) by frequency

Analytical Methods

All ¹H and ¹⁹F NMR spectra were recorded on a Bruker AVANCE NEO 400 NMR operating at 400 MHz for ¹H, and 376 MHz for ¹⁹F respectively. NMR data is recorded in chemical shifts relative to tetramethylsilane (TMS) as internal standard. NMR spectra were run either in CDCl ₃ containing 0.05%TMS, CD ₃OD containing 0.05%TMS, D ₂O or DMSO-d ₆ containing 0.03%TMS.

Preparative HPLC was performed on an Agilent 1260 Infinity II System on Agilent 10 prep-C18 250x21.2 mm column, using an acetonitrile/aqueous 0.1%trifluoroacetic acid gradient, or an acetonitrile/aqueous 0.1%formic acid gradient, or an acetonitrile/water at 22℃.

Mass spectra were performed on an Agilent 1260II-6125 Separation Module using either ES ^-or ES ⁺ ionization modes.

Column Chromatography was performed with using Qingdao Inc. Silica Gel: CC Grade (230 –400 Mesh) .

Commercial solvents and reagents were generally used without further purification. All products were dried before characterisation and use in subsequent synthetic steps.

Example 1

Sodium (2S, 5R) -2-carbamimidoyl-7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 1, Table 1)

Step 1. (2S, 5R) -6- (Benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carbonitrile (III)

A mixture of II (0.2 g, 0.7 mmol) and TEA (0.7 g, 7.0 mmol) in DCM (5 mL) , then TFAA (0.3 g, 1.4 mmol) was added to the reaction mixture at 0 ℃. The resulting reaction mixture was heated at 35 ℃ for 3 hours, and concentrated under reduced pressure. The residue was extracted with ethyl acetate, washed with water, brine, dried over Na ₂SO ₄ and filtrated. The filtrate was concentrated to give a residue, which was further purified by column chromatography eluting with 30%ethyl acetate in hexane to give the title compound III (0.17 g , 64%) as a brown solid.

1H NMR (400 MHz, DMSO-d ₆) : δ 1.81 –1.91 (m, 2 H) , 1.92 –2.00 (m, 2 H) , 3.10 (d, J = 11.7 Hz, 1 H) , 3.21 (d, J = 12.3 Hz, 1 H) , 3.74 (s, 1 H) , 4.58 (d, J = 6.6 Hz, 1 H) , 4.93 (d, J = 11.6 Hz, 1 H) , 4.96 (d, J = 11.6 Hz, 1 H) , 7.36 –7.43 (m, 3 H) , 7.44 –7.48 (m, 3 H) . LC-MS analysis: [M+H] ⁺ = 256.1.

Step 2. tert-Butyl ( ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) (imino) methyl) carbamate (1_1)

AlMe ₃ in n-hexane (2 N, 9.00 mL, 18.0 mmol) and NH ₄Cl (0.96 g, 18.0 mmol) were added to a solution of III (3.08 g, 15.0 mmol) in anhydrous DCM (45 mL) at 0 ℃. The reaction mixture was stirred at room temperature overnight, cooled to 0 ℃, quenched by addition of silica gel (8 g) and methanol (8 mL) . The resulting mixture was stirred at room temperature for 20 min, filtered off, rinsed with 10%methanol in DCM (2x30 mL) . The filtrate was concentrated and purified by flash column chromatography using 2-5%MeOH in DCM to give the title compound 1_1 (1.45 g, 44%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.61 –1.79 (m, 2 H) , 1.80 –1.92 (m, 2 H) , 2.77 –2.95 (m, 2 H) , 3.12 –3.21 (m, 1 H) , 3.80 –3.92 (m, 1 H) , 4.09 –4.15 (m, 1 H) , 4.74 –4.84 (m, 2 H) , 6.52 –6.66 (m, 2 H) , 7.33 –7.42 (m, 3 H) , 7.45 –7.54 (m, 2 H) . LC-MS analysis: [M+Na] ⁺ = 297.1.

Step 3: Synthesis of tert-butyl ( (Z) -amino ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) methylene) carbamate (1_2)

A mixture of crude compound 1_1 (0.5 g, 1.33 mmol) , (Boc) ₂O (0.67 g, 4.0 mmol) and TEA (0.55 mL, 3.0 mmol) in DCM (15 mL) was stirred at 20 ℃ for 48 hours. The reaction mixture was concentrated under reduced pressure and the residue was purified by flash chromatography on silica gel, eluting with 30%EtOAc in petroleum ether to give the title compound 1_2 (0.39 g, 76%) as a white solid.

¹H NMR (400 MHz, CDCl ₃) : δ 1.49 (s, 9 H) , 1.81 –1.91 (m, 1 H) , 1.93 –2.01 (m, 1 H) , 2.04 –2.17 (m, 2 H) , 3.21 (t, J = 11.4 Hz, 1 H) , 4.09 –4.18 (m, 2 H) , 4.89 (s, 2 H) , 5.26 –5.44 (m, 3 H) , 7.38 –7.44 (m, 5 H) . LC-MS analysis: [M+H] ⁺ = 375.2.

Step 4: Synthesis of tert-butyl ( (Z) -amino ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) methylene) carbamate (1_3)

A mixture of compound 1_2 (0.7 g, 1.9 mmol) and Palladium 10%on carbon (wetted with ca. 55%water, 0.2 g, 20%w/w) in MeOH (50 mL) was stirred at 5 ℃ for 10 minutes. Then hydrogen gas was inserted into the system, the mixture was reacted at room temperature overnight. After the reaction mixture was filtrated and the filtrate was concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel, eluting with 70%EtOAc in petroleum ether to give the title compound 1_3 (0.46 g, 71%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.42 (s, 9 H) , 1.66 –1.72 (m, 1 H) , 1.75 –1.82 (m, 2 H) , 1.96 –2.01 (m, 1 H) , 2.80 –2.96 (m, 1 H) , 3.79 –3.93 (m, 2 H) , 5.23 (br s, 1H) , 6.49 (br s, 2 H) , 9.24 (s, 1 H) . LC-MS analysis: [M+Na] ⁺ = 307.2.

Step 5: Synthesis of (2S, 5R) -2- ( (Z) -N’ - (tert-butoxycarbonyl) carbamimidoly) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl) hydrogen sulfate as pyridine salt (1_4)

SO ₃. Pyridine complex (0.56 g, 3.50 mmol) was added to a solution of compound 1_3 (0.20 g, 0.70 mmol) in anhydrous pyridine (7 mL) . The mixture was stirred at room temperature overnight, concentrated to dryness under reduced pressure. The residue was suspended with CH ₂Cl ₂ (10 mL) , filtered off and rinsed with CH ₂Cl ₂ (2x3 mL) . The filtrate was concentrated to give a residue, which was purified by flash column chromatography using 5-15%MeOH in CH ₂CH ₂ to give the title compound 1_4 (0.27 g, 87%) as a pale yellow foam.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.42 (s, 9 H) , 1.65 –1.91 (m, 3 H) , 1.91 –2.03 (m, 1 H) , 2.94 –3.03 (m, 1 H) , 3.77 –3.87 (m, 1 H) , 3.92 –4.00 (m, 1 H) , 5.18 –5.26 (m, 1 H) , 6.44 (br s, 2 H) , 7.98 –8.06 (m, 2 H of pyridine) , 8.50 –8.56 (m, 1 H of pyridine) , 8.88 –8.94 (m, 2 H of pyridine) . LC-MS analysis: [M-Pyr-H] ^-= 363.1.

Step 6: Synthesis of sodium (2S, 5R) -2-carbamimidoyl-7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 1)

TFA (1.5 mL) was added to a solution of compound 1_4 (0.27 g, 0.61 mmol) in anhydrous CH ₂Cl ₂ (8 mL) at 0 ℃. The mixture was stirred for 3.5 hours at 0 ℃, concentrated under reduced pressure to give a residue. The residue was dissolved with CH ₂Cl ₂ (20 mL) and extracted with water (2x10 mL) . The aqueous layer was freeze-dried and purified by resin Dowex-50wx Na ⁺, using water as elution solvent to give example 1 (32 mg, 18%) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.70 –2.05 (m, 4 H) , 3.00 –3.13 (m, 2 H) , 3.77 –3.87 (m, 1 H) , 4.17 –4.27 (m, 1 H) . LC-MS analysis: [M-Na] ^-= 263.1.

Example 2

Sodium (2S, 5R) -2- (N-methylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 2, Table 1)

Step 1: Synthesis of (2S, 5R) -6- (benzyloxy) -N-methyl-7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamide (2_1)

(2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carbonitrile III (0.61 g, 2.3 mmol) was added to a solution of methylamine hydrochloride (0.24 g, 3.5 mmol) in DCM (20 mL) at 0 ℃, and then trimethyl aluminum (1 N in n-hexane, 3.5 mL, 3.5 mmol) was added dropwisely. The mixture was stirred at room temperature under argon overnight. Then 5.0 g silica was added to the reaction mixture under ice cooling bath, and stirred for further 0.5 hour. 10 mL of methanol was added. Then the mixture was filtrated, washed with methanol and EtOAc. The filtrate was concentrated under reduced pressure to give a residue. The residue was dissolved with EtOAc, washed with saturated NH ₄Cl, dried by Na ₂SO ₄ and filtered. The filtrate was concentrated to afford a crude compound, which was purified by flash chromatography using 5%MeOH in DCM to give the title compound 2_1 (0.40 g, 61%) as a white solid.

¹H NMR (400 MHz, CDCl ₃) : δ 1.81 –1.96 (m, 2 H) , 1.97 –2.14 (m, 2 H) , 2.74 (d, J =4.8 Hz, 3 H) , 3.07 –3.14 (m, 1 H) , 3.22 (t, J = 10.9 Hz, 1 H) , 4.06 –4.08 (m, 1 H) , 4.14 –4.22 (m, 1 H) , 4.81 (s, 2 H) , 5.74 (br s, 1 H) , 7.41 –7.44 (m, 5H) . LC-MS analysis: [M+Na] ⁺ =311.2.

Step 2: Synthesis of tert-butyl ( (Z) - ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) (methylamino) methylene) carbamate (2_2)

A mixture of compound 2_1 (0.5 g, 1.73 mmol) , (Boc) ₂O (0.67 g, 4.0 mmol) and TEA (0.55 mL, 3.0 mmol) in DCM (15 mL) was stirred at 20 ℃ for 48 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel eluting with 30%EtOAc in petroleum ether to give the title compound 2_2 (0.4 g, 60%) as a white solid.

¹H NMR (400 MHz, CDCl ₃) : δ 1.49 (s, 9 H) , 1.79 –1.95 (m, 2 H) , 2.02 –2.15 (m, 2 H) , 2.76 (d, J = 4.4 Hz, 3 H) , 3.19 (t, J = 11.1 Hz, 1 H) , 4.06 –4.16 (m, 2 H) , 4.83 (s, 2 H) , 5.76 (s, 1 H) , 7.39 –7.45 (m, 5 H) . LC-MS analysis: [M+H] ⁺ = 411.2.

Step 3: Synthesis of tert-butyl ( (Z) - ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) (methylamino) methylene) carbamate (2_3)

A mixture of compound 2_2 (0.78 g, 2.0 mmol) and palladium 10%on carbon (wetted, 55%water w/w, 0.16 g) in MeOH (50 mL) was stirred at 5 ℃ for ten minutes. Then hydrogen gas was inserted into the system, the mixture was reacted at room temperature overnight. After the reaction mixture was filtrated and the filtrate was concentrated under reduced pressure to give the title compound 2_3 (0.55 g, 66%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.42 (s, 9 H) , 1.64 –1.69 (m, 1 H) , 1.74 –1.82 (m, 2 H) ,1.95 –2.01 (m, 1 H) , 2.61 (d, J = 4.46 Hz, 3 H) , 3.17 (d, J = 5.09 Hz, 1 H) , 3.77 –3.91 (m, 2 H) , 5.22 (br s, 1 H) , 6.94 –6.99 (m, 2 H) , 9.15 (s, 1 H) . LC-MS analysis: [M+Na] ⁺ = 321.2.

Step 4: Synthesis of (2S, 5R) -2- ( (Z) -N’ - (tert-butoxycarbonyl) -N-methylcarbamimidoly) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl) hydrogen sulfate pyridine salt (2_4)

SO ₃. Pyridine complex (0.51 g, 3.19 mmol) was added to a solution of compound 2_3 (0.16 g, 0.53 mmol) in anhydrous pyridine (6 mL) . The mixture was stirred at room temperature overnight. The reaction mixture was concentrated to dryness under reduced pressure. The residue was suspended with CH ₂Cl ₂ (10 mL) , filtered off and rinsed with CH ₂Cl ₂ (2x3 mL) . The filtrate was concentrated and purified by flash column chromatography using 5-10%MeOH in CH ₂CH ₂ to give the title compound 2_4 (0.19 g, 79%) as a white foam.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.43 (s, 9 H) , 1.64 –1.89 (m, 3 H) , 1.91 –2.00 (m, 1 H) , 2.63 (d, J = 4.50 Hz, 3 H) , 2.96 –3.13 (m, 1 H) , 3.76 –3.86 (m, 1 H) , 3.90 –4.00 (m, 1 H) , 5.18 –5.26 (m, 1 H) , 6.75 (d, J = 4.50 Hz, 1 H) , 7.85 –7.92 (m, 2 H of pyridine) , 8.33 –8.41 (m, 1 H of pyridine) , 8.80 –8.87 (m, 2 H of pyridine) . LC-MS analysis: [M-Pyr-H] ^-= 377.1.

Step 5: Synthesis of sodium (2S, 5R) -2- (N-methylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 2)

TFA (1.2 mL) was added to a solution of compound 2_4 (0.19 g, 0.42 mmol) in anhydrous CH ₂Cl ₂ (8 mL) at 0 ℃. The mixture was stirred at 0 ℃ for 3 hours. The reaction mixture was concentrated under reduced pressure. The residue was dissolved with CH ₂Cl ₂ (20 mL) and extracted with water (2x10 mL) . The aqueous layer was freeze-dried and purified by resin Dowex-50wx Na ⁺ using water to give example 2 (26 mg, 21%) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.69 –2.04 (m, 4 H) , 2.63 (s, 3 H) , 3.00 –3.10 (m, 2 H) , 3.83 –3.93 (m, 1 H) , 4.19 –4.24 (m, 1 H) . LC-MS analysis: [M-Na] ^-= 277.1.

Example 3

(2S, 5R) -2- (N-ethylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate (compound 3, Table 1)

Step 1: Synthesis of (2S, 5R) -6- (benzyloxy) -N-ethyl-7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamide (3_1)

To a solution of (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carbonitrile III (0.60 g, 2.3 mmol) in DCM (20 mL) at 0 ℃ was added ethylamine hydrochloride (0.29 g, 3.5 mmol) and then added trimethyl aluminum (3.5 mL, 1 N in hexane) dropwisely. The mixture was stirred at room temperature under argon overnight. Then 5.0 g silica was added under ice cooling bath, the mixture stirred for further 0.5 hour, 10 mL of methanol was added. Then the mixture was filtrated, washed with methanol and EtOAc, concentrated under reduced pressure to give a residue. The residue was dissolved with EtOAc, washed with saturated NH ₄Cl, dried by Na ₂SO ₄, concentrated to provide the crude compound 3_1 (0.5 g) , which was directly used for next step without further purification. LC-MS analysis: [M+Na] ⁺ = 325.1.

Step 2: Synthesis of tert-butyl ( (Z) - ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) (ethylamino) methylene) carbamate (3_2)

A mixture of crude compound 3_1 (0.5 g) , (Boc) ₂O (0.67 g, 4.0 mmol) and TEA (0.55 mL, 3.0 mmol) in DCM (15 mL) was stirred at 20 ℃ for 48 hours. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel, eluting with 30%EtOAc in petroleum ether to provide the title compound 3_2 (0.41 g, 44%in two steps) as a white solid.

¹H NMR (400 MHz, CDCl ₃) : δ 1.00 (t, J = 7.1 Hz, 3 H) , 1.45 (s, 9 H) , 1.75 –1.94 (m 2 H) , 1.97 –2.13 (m, 2 H) , 3.10 –3.22 (m, 3 H) , 4.08 (q, J = 7.1 Hz, 2 H) , 4.81 (s, 2 H) , 5.21 (br s, 1 H) , 5.62 –5.95 (m, 1 H) , 7.41 (s, 5 H) . LC-MS analysis: [M+H] ⁺ = 403.1.

Step 3: Synthesis of tert-butyl ( (Z) - (ethylamino) ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) methylene) carbamate (3_3)

A mixture of compound 3_2 (1.1 g, 2.7 mmol) and palladium 10%on carbon (wetted with ca. 55%water, 0.2 g, 20%rel-weight) in MeOH (50 mL) was stirred at 5 ℃ for ten minutes. Then hydrogen gas was inserted into the system, the mixture was reacted at room temperature overnight. The reaction mixture was filtrated and concentrated under reduced pressure to provide a residue, which was purified by flash chromatography on silica gel, eluting with 70%EtOAc in petroleum ether to give the title compound 3_3 (0.55 g, 66%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.01 (t, J = 7.1 Hz, 3H) , 1.43 (s, 9H) , 1.63 –1.70 (m, 1 H) , 1.74 –1.82 (m, 2 H) , 1.95 –2.01 (m, 1 H) , 2.79 –2.94 (m, 1 H) , 3.03 –3.12 (m, 2H) , 3.78 –3.91 (m, 2 H) , 5.01 –5.42 (m, 1 H) , 7.11 (t, J = 5.9 Hz, 1 H) , 9.19 (s, 1 H) . LC-MS analysis: [M+Na] ⁺ = 335.1.

Step 4: Synthesis of (2S, 5R) -2- ( (Z) -N'- (tert-butoxycarbonyl) -N-ethylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate (3_4)

A mixture of compound 3_3 (0.35 g, 1.12 mmol) , SO ₃. NMe ₃ complex (0.47 g, 3.36 mmol) and TEA (0.47 mL, 3.36 mmol) in MeOH (15 mL) was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to provide a residue, which was washed with a small amount of water to give a solid, which was collected and dried to give the crude compound 3_4 as triethylamine salt (0.41 g, 74%) as a solid, which was used for next step without further purification. LC-MS analysis: [M-H] ^-= 391.1.

Step 5: Synthesis of (2S, 5R) -2- (N-ethylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate (example 3)

The crude compound 3_4 (0.07 g, 0.14 mmol) was added into formic acid (3.0 mL, 88%) was stirred at 0 ℃ for 30 hours. The reaction mixture was concentrated under reduced pressure to provide residue. The residue was dissolved with water, and then neutralized by saturated NaHCO ₃. The solution was lyophilized and then purified by preparative HPLC on an Agilent 10 prep-C18 250x21.2 mm column and lyophilized to give example 3 (15 mg, 36%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.01 (t, J = 7.2 Hz, 3H) , 1.71 –1.96 (m, 2 H) , 2.08 –2.16 (m, 1 H) , 2.22 –2.34 (m, 1 H) , 3.04 –3.33 (m, 3 H) , 4.22 (br s, 1 H) , 4.85 (br s, 1 H) , 6.82 –6.89 (m, 1 H) . LC-MS analysis: [M-H] ^-= 291.1.

Example 4

Sodium (2S, 5R) -2- (N- (2-acetamidoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 10, Table 1)

Step 1: Synthesis of N- (2- ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) ethylacetamide (4_1)

Trifluoromethanesulfonate (TMSOTf, 0.60 mL, 3.31 mmol) and N- (2-aminoethyl) acetamide (1.02 g, 10.00 mmol) were added to a solution of III (1.67 g, 6.50 mmol) in anhydrous THF (20 mL) at 0 ℃. The reaction mixture was stirred at room temperature for 3.5 days, concentrated, diluted with CH ₂Cl ₂ (80 mL) , washed with water (20 mL) , brine (15 mL) and dried over Na ₂SO ₄. The filtrate was concentrated and purified by flash column chromatography using 2-5%MeOH in CH ₂CH ₂ to give the title compound 4_1 (1.11 g, 47%) as a white foam.

¹H NMR (400 MHz, CDCl ₃) : δ 1.77 –1.86 (m, 2 H) , 1.94 (s, 3 H) . 1.92 –2.01 (m, 1 H) , 2.11–2.20 (m, 1 H) , 2.79 –2.88 (m, 1 H) , 3.10 –3.16 (m, 1 H) , 3.19 –3.29 (m, 4 H) , 3.52 –3.60 (m, 1 H) , 3.96 –4.06 (m, 1 H) , 4.76 (s, 2 H) , 6.06 (br s, 2 H) , 7.32 –7.45 (m, 5 H) . LC-MS analysis: [M+H] ⁺ = 360.1.

Step 2: Synthesis of tert-butyl ( (Z) - ( (2-acetamidoethyl) amino ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) methylene) carbamate (4_2)

TEA (0.70 mL, 5.00 mmol) and (Boc) ₂O (0.70 g, 3.21 mmol) were added to a solution of compound 4_1 (1.10 g, 3.06 mmol) in CH ₂Cl ₂ (15 mL) at 0 ℃. The resulting mixture was stirred at room temperature overnight, concentrated and purified by flash column chromatography using EtOAc to give the title compound 4_2 (0.98 g, 70%) as a white foam.

¹H NMR (400 MHz, CDCl ₃) : δ 1.36 (s, 9 H) , 1.80 –1.90 (m, 1 H) , 1.95 (s, 3 H) , 2.00 –2.08 (m, 2 H) , 2.30 –2.3 (m, 1 H) , 3.09 –3.20 (m, 1 H) , 3.21 –3.42 (m, 4 H) , 4.23 –4.39 (m, 2 H) , 4.69 –4.79 (m, 2 H) , 5.23 (s, 1 H) , 6.00 –6.10 (m, 1 H) , 7.28 –7.44 (m, 5 H) . LC-MS analysis: [M+H] ⁺ = 460.1.

Step 3: Synthesis of tert-butyl ( (Z) - ( (2-acetamidoethyl) amino ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) methylene) carbamate (4_3)

10%Pd/C (wet, 55%water w/w, 0.11 g) was added to a solution of compound 4_2 (0.97 g, 2.11 mmol) in EtOAc (10 mL) . The mixture was stirred under H ₂ (balloon) overnight, filtered through a pad of celite, rinsed with EtOAc (3x10 mL) . The filtrate was concentrated to give the title compound 4_3 (0.71 g, 94%) as a white solid, which was directly used for next step without further purification.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.41 (s, 9 H) , 1.72 –1.92 (m, 3 H) , 1.80 (s, 3 H) , 2.14 –2.24 (m, 1 H) , 3.04 –3.14 (m, 4 H) , 3.16 –3.24 (m, 1 H) , 3.79 –3.90 (m, 1 H) , 4.04 –4.13 (m, 1 H) , 5.05 –5.12 (m, 1 H) , 7.03 –7.10 (m, 1 H) , 7.81 –7.92 (m, 1 H) , 9.14 (s, 1 H) . LC-MS analysis: [M+H] ⁺ =370.1

Step 4: Synthesis of (2S, 5R) -2- ( (Z) -N- (2-acetamidoethyl) -N’ - (tert-butoxycarbonyl) carbamimidoly) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl) hydrogen sulfate as pyridine salt (4_4)

SO ₃. Pyridine complex (0.64 g, 4.00 mmol) was added to a solution of compound 4_3 (0.29 g, 0.79 mmol) in anhydrous pyridine (7 mL) . The mixture was stirred at room temperature overnight, concentrated to dryness under reduced pressure to give a residue. The residue was suspended with CH ₂Cl ₂ (15 mL) , filtered off and rinsed with CH ₂Cl ₂ (2x5 mL) . The filtrate was concentrated and purified by flash column chromatography using 5-15%MeOH in CH ₂CH ₂ to give the title compound 4_4 (0.37 g, 90%) as a pale yellow foam.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.41 (s, 9 H) , 1.67 –1.88 (m, 2 H) , 1.78 (s, 3 H) , 2.10 –2.33 (m, 2 H) 3.03 –3.15 (m, 4 H) , 3.23 –3.33 (m, 1 H) , 3.72 –3.83 (m, 1 H) , 4.04 –4.14 (m, 1 H) , 4.94 –5.05 (m, 1 H) , 6.93 –7.03 (m, 1 H) , 7.69 –7.79 (m, 1 H) , 7.91 –8.00 (m, 2 H of pyridine) , 8.42 –8.50 (m, 1 H of pyridine) , 8.82 –8.91 (m, 2 H of pyridine) . LC-MS analysis: [M-Pyr-H] ^-= 448.2.

Step 5: Synthesis of sodium (2S, 5R) -2- (N- (2-acetamidoethyl) carbamimidoly) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 4)

TFA (1.4 mL) was added to a solution of compound 4_4 (0.37 g, 0.70 mmol) in anhydrous CH ₂Cl ₂ (9 mL) at 0 ℃. The mixture was stirred for 3.5 hours at 0 ℃, and concentrated under reduced pressure to give a residue. The residue was dissolved with CH ₂Cl ₂ (20 mL) and extracted with water (2x10 mL) . The aqueous layer was freeze-dried and purified by resin Dowex-50wx Na ⁺ using water as an eluting solvent to afford example 4 (40 mg, 15%) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.62 –1.75 (m, 1 H) , 1.82 (s, 3 H) , 1.82 –1.92 (m, 2 H) , 2.16 –2.26 (m, 1 H) , 3.11 –3.26 (m, 5 H) , 3.33 –3.45 (m, 1 H) , 3.68 –3.77 (m, 1 H) , 4.09 –4.21 (m, 1 H) . LC-MS analysis: [M-Na] ^-= 348.2.

Example 5

Sodium (2S, 5R) -2- (N- (1- (ethoxycarbonyl) piperidin-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 59, Table 1)

Step 1: Synthesis of ethyl 4- ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) piperidine-1-caboxylate (5_1)

TMSOTf (0.50 mL, 2.76 mmol) and ethyl 4-aminopiperidine-1-carboxylate (1.40 mL, 8.17 mmol) were added to a solution of III (1.30 g, 5.00 mmol) in anhydrous THF (20 mL) at 0 ℃. The reaction mixture was stirred at room temperature for 3.5 days, concentrated, diluted with EtOAc (80 mL) , washed with water (20 mL) , brine (15 mL) and dried over Na ₂SO ₄. The filtrate was concentrated and purified by flash column chromatography using 50%EtOAc in petroleum ether to give the title compound 5_1 (0.56 g, 26%) as a white foam.

¹H NMR (400 MHz, CDCl ₃) : δ 1.02 –1.07 (m, 2 H) , 1.27 (t, J = 7.07 Hz, 3 H) , 1.68 –1.87 (m, 5 H) , 1.95 –2.08 (m, 1 H) , 2.80 –2.93 (m, 2 H) , 3.22 –3.32 (m, 2 H) , 3.65 –3.76 (m, 1 H) , 3.81 –4.03 (m, 3 H) , 4.14 (q, J = 7.07 Hz, 2 H) , 4.67 –4.74 (m, 2 H) , 4.82 (d, J = 7.10 Hz, 1 H) , 5.40-5.60 (m, 2 H) , 7.28 –7.38 (m, 5 H) . LC-MS analysis: [M+H] ⁺ = 430.2.

Step 2: Synthesis of ethyl 4- ( (2S, 5R, E) -6- (benzyloxy) -7-oxo-N’ - (2, 2, 2-trifluoroacetyl) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) piperidine-1-caboxylate (5_2)

TEA (0.65 mL, 4.65 mmol) and TFAA (0.33 mL, 2.33 mmol) were added to a solution of compound 5_1 (0.80 g, 1.86 mmol) in CH ₂Cl ₂ (15 mL) at 0 ℃. The resulting mixture was stirred at room temperature for 4 hours. The reaction mixture was concentrated to afford a residue, which was purified by flash column chromatography using 25%EtOAc in hexane to give the title compound 5_2 (0.35 g, 36%) as a white foam.

¹H NMR (400 MHz, CDCl ₃) : δ 1.07 –1.30 (m, 2 H) , 1.26 (t, J = 7.10 Hz, 3 H) , 1.65 –1.75 (m, 1 H) , 1.83 –1.97 (m, 2 H) , 2.02 –2.14 (m, 2 H) , 2.32 –2.42 (m, 1 H) , 2.77 –2.94 (m, 2 H) , 3.41 –3.50 (m, 1 H) , 3.63 –3.76 (m, 1 H) , 3.91 –4.09 (m, 2 H) , 4.12 (q, J = 7.10 Hz, 2 H) , 4.27 –4.41 (m, 2 H) , 4.82 –4.90 (m, 1 H) , 4.95 (s, 2 H) , 5.30 –5.37 (m, 1 H) , 7.30 –7.45 (m, 5 H) ; ¹⁹F NMR (400 MHz, CDCl ₃) : δ -70.98 (s, 3F) . LC-MS analysis: [M+H] ⁺ = 526.2.

Step 3: Synthesis of ethyl 4- ( (2S, 5R, E) -6-hydroxy-7-oxo-N’ - (2, 2, 2-trifluoroacetyl) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) piperidine-1-caboxylate (5_3)

10%Pd/C (0.120 g) was added to a solution of compound 5_2 (0.35 g, 0.66 mmol) in EtOAc (12 mL) . The mixture was stirred under H ₂ (balloon) overnight, filtered through a pad of celite, rinsed with EtOAc (3x10 mL) . The filtrate was concentrated to afford a residue, which was purified by flash column chromatography using 50%EtOAc in petroleum ether to give the title compound 5_3 (0.18 g, 61%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.18 (t, J = 7.10 Hz, 3 H) , 1.22 –1.34 (m, 2 H) , 1.64 –1.77 (m, 2 H) , 1.79 –1.92 (m, 2 H) , 1.95 –2.03 (m, 1 H) , 2.10 –2.20 (m, 1 H) , 2.73 –2.93 (m, 2 H) , 3.18 –3.28 (m, 1 H) , 3.57 –3.69 (m, 1 H) , 3.84 –3.96 (m, 3 H) , 4.02 (q, J = 7.10 Hz, 2 H) , 4.31 –4.40 (m, 1 H) , 5.20 –5.27 (m, 1 H) , 6.65 (d, J = 7.20 Hz, 1 H) , 10.6 (s, 1 H) ; ¹⁹F NMR (400 MHz, DMSO-d ₆) : δ -70.00 (s, 3F) . LC-MS analysis: [M+H] ⁺ =434.2.

Step 4: Synthesis of ethyl 4- ( (2S, 5R, E) -7-oxo-6- (sulfooxy) -N’ - (2, 2, 2-trifluoroacetyl) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) piperidine-1-caboxylate as pyridine salt (5_4)

SO ₃. Pyridine complex (0.32 g, 2.00 mmol) was added to a solution of compound 5_3 (0.18 g, 0.40 mmol) in anhydrous pyridine (4 mL) . The mixture was stirred overnight and concentrated to dryness under reduced pressure to afford a residue. The residue was suspended with CH ₂Cl ₂ (10 mL) , filtered off and rinsed with CH ₂Cl ₂ (2x3 mL) . The filtrate was concentrated and purified by flash column chromatography using 5-10%MeOH in CH ₂CH ₂ to give the title compound 5_4 (0.19 g, 78%) as a pale yellow foam.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.18 (t, J = 7.10 Hz, 3 H) , 1.21 –1.34 (m, 4 H) , 1.66 –1.88 (m, 4 H) , 2.78 –2.96 (m, 2 H) , 3.22 –3.32 (m, 1 H) , 3.54 –3.66 (m, 1 H) , 3.81 –3.92 (m, 2 H) , 4.02 (q, J = 7.10 Hz, 2 H) , 4.23 –4.33 (m, 2 H) , 5.04 –5.09 (m, 1 H) , 6.44 (d, J =7.20 Hz, 1 H) , 7.88-7.97 (m, 2 H of pyridine) , 8.39 –8.47 (m, 1 H of pyridine) , 8.82 –8.90 (m, 2 H of pyridine) (SO ₃H missing) ; ¹⁹F NMR (400 MHz, DMSO-d ₆) : δ -68.72 (s, 3F) . LC-MS analysis: [M-Pyr-H] ^-= 514.2.

Step 5: Synthesis of sodium (2S, 5R) -2- (N- (1-ethylcarbonyl) piperidin-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 5)

Na ₂CO ₃ (65.0 mg, 0.610 mmol) was added to a solution of compound 5_4 (0.19 g, 0.31 mmol) in MeOH and H ₂O (4 mL and 3 mL) . The mixture was stirred for 2 hours at room temperature. The reaction mixture was concentrated to afford a residue which was purified by Diaion HP-20, using water and then 10%CH ₃CN in water as eluting solvent to provide example 5 (13 mg, 10%) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.17 (t, J = 7.17 Hz, 3 H) , 1.57 –1.72 (m, 3 H) , 1.76 –1.88 (m, 1 H) , 1.93 –2.22 (m, 4 H) , 2.69 –2.92 (m, 3 H) , 2.98 –3.14 (m, 1 H) , 3.92 –4.20 (m, 5 H) 4.05 (q, J = 7.17 Hz, 2 H) (2 NH not shown) . LC-MS analysis: [M-Na] ^-= 418.2.

Example 6

Sodium (2S, 5R) -7-oxo-2- (N- (pyridin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 91, Table 1)

Step 1: Synthesis of (2S, 5R) -6- (benzyloxy) -7-oxo-N- (pyridin-3-ylmethyl) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamide (6_1)

AlMe ₃ in n-hexane (2 N, 4.50 mL, 9.00mmol) and pyridine-3-ylmethanamine (0.96 g, 8.88 mmol) were added to a solution of III (1.58 g, 6.15mmol) in anhydrous CH ₂Cl ₂ (18 mL) at 0 ℃. The reaction mixture was stirred at room temperature for 40 hours, cooled to 0 ℃, quenched by addition of silica gel (5 g) and methanol (5 mL) . The resulting mixture was stirred at room temperature for 20 min, filtered off, rinsed with 10%methanol in CH ₂CH ₂ (2x30 mL) . The filtrate was concentrated to afford a residue, which was purified by flash column chromatography using 1-3%MeOH in CH ₂CH ₂ to give the title compound 6_1 (0.94 g, 38%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.62 –1.78 (m, 2 H) , 1.81 –1.92 (m, 2 H) , 2.76 –2.81 (m, 1 H) , 2.88 –2.97 (m, 1 H) , 3.13 –3.21 (m, 1 H) , 3.80 –3.91 (m, 1 H) , 4.08 –4.15 (m, 1 H) , 4.28 (d, J = 6.0 Hz, 2 H) , 4.77 –4.86 (m, 2 H) , 7.29 –7.40 (m, 4 H) , 7.46 –7.53 (m, 2 H) , 7.57 –7.62 (m, 1 H) , 7.76 (t, J = 6.0 Hz. 1 H) , 8.41 –8.47 (m, 2 H) . LC-MS analysis: [M+H] ⁺ = 366.2.

Step 2: Synthesis of tert-butyl ( (Z) - ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) ( (pyridin-3-ylmethyl) amino) methylene) carbamate (6_2)

TEA (0.65 mL, 4.64 mmol) and (Boc) ₂O (0.85 g, 3.87 mmol) were added to a solution of 6_1 (0.94 g, 2.58 mmol) in anhydrous CH ₂Cl ₂ (12 mL) at 0 ℃. The reaction mixture was stirred at room temperature for 24 hours, concentrated to afford a residue, which was purified by flash column chromatography using EtOAc to give the title compound 6_2 (0.60 g, 50%) as a pale yellow foam.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.40 (s, 9 H) , 1.74 –1.92 (m, 3 H) , 1.94 –2.02 (m, 1 H) , 2.86 –2.98 (m, 1 H) , 3.78 –3.91 (m, 2 H) , 4.25 –4.35 (m, 2 H) , 4.80 –4.89 (m, 2 H) , 5.21 –5.26 (m, 1 H) , 7.30 –7.39 (m, 4 H) , 7.45 –7.50 (m, 2 H) , 7.58 –7.63 (m, 1 H) , 7.88 (t, J = 6.0 Hz. 1 H) , 8.42 –8.49 (m, 2 H) . LC-MS analysis: [M+H] ⁺ = 466.3.

Step 3: Synthesis of tert-butyl ( (Z) - ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) ( (pyridin-3-ylmethyl) amino) methylene) carbamate (6_3)

10%Pd/C (0.160 g) was added to a solution of compound 6_2 (0.41 g, 0.86 mmol) in MeOH (16 mL) . The mixture was stirred under H ₂ (balloon) for 7 hours, filtered through a pad of celite, rinsed with MeOH (2x5 mL) . The filtrate was concentrated and purified by flash column chromatography using 3-5%MeOH in CH ₂CH ₂to give the title compound 6_3 (0.10 g, 33%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.41 (s, 9 H) , 1.65 –1.72 (m, 1 H) , 1.74 –1.82 (m, 2 H) , 1.95 –2.02 (m, 1 H) , 2.83 –2.96 (m, 1 H) , 3.78 –3.91 (m, 2 H) , 4.24 –4.30 (m, 2 H) , 4.80 –4.89 (m, 2 H) , 5.21 –5.25 (m, 1 H) , 7.30 –7.36 (m, 1 H) , 7.61 –7.67 (m, 1 H) , 7.78 (t, J = 6.0 Hz. 1 H) , 8.40 –8.49 (m, 2 H) , 9.33 (s, 1 H) . LC-MS analysis: [M+H] ⁺ =376.2.

Step 4: Synthesis of (2S, 5R) -2- ( (Z) -N’ - (tert-butoxycarbonyl) -N- (pyridin-3-ylmethyl) carbamimidoly) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl) hydrogen sulfate as pyridine salt (6_4)

SO ₃. Pyr complex (0.50 g, 3.14 mmol) was added to a solution of compound 6_3 (0.21 g, 0.53 mmol) in anhydrous pyridine (6 mL) . The mixture was stirred overnight, concentrated to dryness under reduced pressure. The residue was suspended with CH ₂Cl ₂ (10 mL) , filtered off and rinsed with CH ₂Cl ₂ (2x3 mL) . The filtrate was concentrated and purified by flash column chromatography using 5-15%MeOH in CH ₂CH ₂ to give the title compound 6_4 (0.29 g, quantitative) as a pale yellow foam.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.41 (s, 9 H) , 1.69 –1.83 (m, 2 H) , 1.85 –2.01 (m, 2 H) , 3.07 –3.12 (m, 1 H) , 3.77 –3.86 (m, 1 H) , 3.94 –4.00 (m, 1 H) , 4.48 –4.56 (m, 2 H) , 5.18 –5.26 (m, 1 H) , 7.63 (t, J = 6.0 Hz, 1 H) , 7.97 –8.06 (m, 3 H, including 2 H of pyridine) , 8.47 –8.55 (m, 2 H, including 1 H of pyridine) , 8.77 –8.92 (m, 4 H, including 2 H of pyridine) . LC-MS analysis: [M-Pyr-H] ^-= 454.2.

Step 5: Synthesis of sodium (2S, 5R) -7-oxo-2- (N- (pyridin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 6)

TFA (1.5 mL) was added to a solution of compound 6_4 (0.29 g, 0.53 mmol) in anhydrous CH ₂Cl ₂ (6 mL) at 0 ℃. The mixture was stirred for 3.5 hours at 0 ℃, concentrated under reduced pressure to afford a residue. The residue was dissolved with CH ₂Cl ₂ (20 mL) and extracted with water (2x10 mL) . The aqueous layer was freeze-dried and purified by resin Dowex-50wx Na ⁺, using water as an elution solvent to give example 6 (0.01 g, 5%) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.64 –1, 78 (m, 2 H) , 1.81 –1.94 (m, 2 H) , 2.91 –2.98 (m, 2 H) , 3.77 –3.84 (m, 1 H) , 4.02 –4.05 (m, 1 H) , 4.31 (s, 2 H) , 7.25 –7.30 (m, 1 H) , 7.63 –7.68 (m, 1 H) , 8.25 –8.29 (m, 1 H) , 8.30 –8.34 (m, 1 H) . LC-MS analysis: [M-Na] ^-= 355.2.

Example 7

Sodium (2S, 5R) -2- (N-cyclopropylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 37, Table 1)

Step 1: Synthesis of (2S, 5R) -6- (benzyloxy) -N-cyclopropyl-7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamide (7_1)

AlMe ₃ in n-hexane (2 N, 1.50 mL, 3.00 mmol) and cyclopropylamine (200 uL, 3.0 mmol) were added to a solution of III (0.5 g, 2.00 mmol) in anhydrous CH ₂Cl ₂ (20 mL) at 0 ℃. The reaction mixture was stirred at room temperature overnight, cooled to 0 ℃, quenched by addition of silica gel (2.5 g) and methanol (10 mL) . The resulting mixture was stirred at room temperature for 20 min, filtered off, rinsed with 10%methanol in CH ₂CH ₂. The filtrate was concentrated to afford a residue. The residue was dissolved with EtOAc, washed with saturated NH ₄Cl, dried by NaSO ₄, concentrated to give the crude title compound 7_1 (0.5 g) , which was directly used for next step without further purification. LC-MS analysis: [M+Na] ⁺ =337.1.

Step 2: Synthesis of tert-butyl ( (E) - ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) (cyclopropylamino) methylene) carbamate (7_2)

TEA (2.0 mL, 15 mmol) and (Boc) ₂O (2.0 g, 10.0 mmol) were added to a solution of 7_1 (0.5 g obtained above) in anhydrous CH ₂Cl ₂ (30 mL) at 0 ℃. The reaction mixture was stirred at room temperature for 48 hours, concentrated to afford a residue, which was purified by flash column chromatography using EtOAc and petroleum ether to give the title compound 7_2 (0.45 g, 54%in two steps) as a white solid.

¹H NMR (400 MHz, CDCl ₃) δ 0.43 –1.02 (m, 4 H) 1.23 (s, 9 H) , 1.50 –1.88 (m, 3 H) , 2.05 –2.52 (m, 2 H) , 3.15 (dd, J = 4.5, 15.3 Hz, 1 H) , 3.88 –4.13 (m, 2 H) , 4.39 –4.66 (m, 2 H) , 5.05 (s, 1 H) , 5.43 –5.71 (m, 1 H) , 6.94 –7.33 (m, 5 H) . LC-MS analysis: [M+H] ⁺ = 414.1. Step 3: Synthesis of tert-butyl ( (E) - (cyclopropylamino) ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) methylene) carbamate (7_3)

10%Pd/C (0.2 g, 55%water) was added to a solution of compound 7_2 (0.45 g, 1.08 mmol) in EtOAc (20 mL) . The mixture was stirred under H ₂ (balloon) overnight, filtered through a pad of celite, rinsed with EtOAc. The filtrate was concentrated and purified by flash column chromatography using 70%EtOAc in petroleum ether to give the title compound 7_3 (0.30 g, 85%) as a white solid. LC-MS analysis: [M+Na] ⁺ =347.1.

Step 4: Synthesis of (2S, 5R) -2- ( (E) -N'- (tert-butoxycarbonyl) -N-cyclopropylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate trimethyl amaine salt (7_4)

SO ₃. NMe ₃ complex (0.20 g, 1.43 mmol) was added to a solution of compound 7_3 (0.30 g, 0.93 mmol) and TEA (2 mL) in THF/water (each 10 mL) . The mixture was stirred overnight, concentrated to dryness under reduced pressure to give the title compound 7_4 (0.53 g, crude) as a pale yellow foam. LC-MS analysis: [M-H] ^-=403.2.

Step 5: Synthesis of sodium (2S, 5R) -2- (N-cyclopropylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 7)

TFA (0.5 mL) was added to a solution of compound 7_4 (66 mg obtained above) in anhydrous CH ₂Cl ₂ (2 mL) at 0 ℃. The mixture was stirred for 3.5 hours at 0 ℃, concentrated under reduced pressure to afford a residue. The residue was dissolved with CH ₂Cl ₂ (5 mL) and extracted with water (2x5 mL) . The aqueous layer was freeze-dried and purified by Dowex-50wx Na ⁺ resin, using water as an elution solvent to give example 7 (3 mg, 8%in two steps) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.67 –1.91 (m, 6 H) , 2.05 –2.21 (m, 2 H) , 2.45 –2.51 (m, 1 H) , 2.94 –3.05 (m, 2 H) , 3.80 –3.90 (m, 1 H) , 4.09 –4.11 (m, 1 H) . LC-MS analysis: [M-Na] ^-= 303.1

Example 8

Sodium (2S, 5R) -7-oxo-2- (N- ( (tetrahydro-2H-pyran-4-yl) methyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 89, Table 1)

Step 1: Synthesis of (2S, 5R) -6- (benzyloxy) -7-oxo-N- ( (tetrahydro-2H-pyran-4-yl) methyl) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamide (8_1)

AlMe ₃ in n-hexane (2 N, 2.7 mL, 5.40 mmol) was added to a solution of III (1.0 g, 3.60 mmol) and (tetrahydro-2H-pyran-4-yl) methanamine (0.64 g, 5.40 mmol) in anhydrous CH ₂Cl ₂ (30 mL) at 0 ℃. The reaction mixture was stirred at room temperature overnight, cooled to 0 ℃, quenched by addition of silica gel (3 g) and methanol (15 mL) . The resulting mixture was stirred at room temperature for 20 min, filtered off, rinsed with 10%methanol in CH ₂CH ₂. The filtrate was concentrated to afford a residue. The residue was dissolved with EtOAc, washed with saturated NH ₄Cl, dried by NaSO ₄, concentrated to give the crude title compound 8_1 (1.37 g) , which was directly used for next step without further purification. LC-MS analysis: [M+Ha] ⁺ = 373.2.

Step 2: Synthesis of tert-butyl ( (E) - ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) ( ( (tetrahydro-2H-pyran-4-yl) methyl) amino) methylene) carbamate (8_2)

TEA (1.3 mL, 7.5 mmol) and (Boc) ₂O (0.82 g, 3.8 mmol) were added to a solution of 8_1 (0.7 g obtained above) in anhydrous CH ₂Cl ₂ (30 mL) at 0 ℃. The reaction mixture was stirred at room temperature for 3 days, concentrated to afford a residue, which was purified by flash column chromatography using EtOAc and petroleum ether to give the title compound 8_2 (0.35 g, 54%in two steps) as a white solid.

¹H NMR (400 MHz, CDCl ₃) : δ 1.39 –1.44 (m, 12 H) 1.49 –1.61 (m, 2 H) , 1.75 –1.94 (m, 2 H) , 1.99 –2.14 (m, 2 H) , 2.95 –3.60 (m, 2 H) , 3.13 –3.22 (m, 1 H) , 3.26 –3.33 (m, 2 H) , 3.89-3.95 (m, 2 H) , 4.63 –4.13 (m, 2 H) , 4.81 (s, 2 H) , 5.83 (s, 1 H) , 7.36 –7.41 (m, 5 H) . LC-MS analysis: [M+Na] ⁺ = 495.1.

Step 3: Synthesis of tert-butyl ( (E) - ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) ( ( (tetrahydro-2H-pyran-4-yl) methyl) amino) methylene) carbamate (8_3)

10%Pd/C (0.1 g, 55%water) was added to a solution of compound 8_2 (0.30 g, 0.63 mmol) in MeOH (30 mL) . The mixture was stirred under H ₂ (balloon) overnight, filtered through a pad of celite, rinsed with EtOAc. The filtrate was concentrated to give the title compound 8_3 (0.20 g, 82%) as a white solid, which was directly used for next step without further purification. LC-MS analysis: [M+H] ⁺ =383.2.

Step 4: Synthesis of tert-butyl ( (E) - ( (2S, 5R) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octan-2-yl) ( ( (tetrahydro-2H-pyran-4-yl) methyl) amino) methylene) carbamate (8_4)

SO ₃. NMe ₃ complex (0.11 g, 0.78 mmol) was added to a solution of compound 8_3 (0.20 g, 0.52 mmol) and TEA (1 mL) in THF and water (each 10 mL) . The mixture was stirred overnight, concentrated to dryness under reduced pressure to give a residue, which was purified by Dowex-50wx Na ⁺ resin, using water as an elution solvent to give the desired compound 8_4 (198 mg, 81%) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.13 –1.24 (m, 2 H) , 1.38 (s, 9 H) , 1.56 –1.62 (m, 2 H) , 1.70 –1.81 (m, 2 H) , 1.88 –2.06 (m, 3 H) , 2.99 –3.15 (m, 3 H) , 3.36 (t, J = 11.4 Hz, 2 H) , 4.03 –4.11 (m, 1 H) , 3.81 –3.92 (m, 2 H) , 5.28 (s, 1 H) . LC-MS analysis: [M-Na] ^-= 461.2

Step 5: Synthesis of sodium (2S, 5R) -7-oxo-2- (N- ( (tetrahydro-2H-pyran-4-yl) methyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 8)

TFA (0.5 mL) was added to a solution of compound 8_4 (70 mg, 0.15 mmol) in anhydrous CH ₂Cl ₂ (2 mL) at 0 ℃. The mixture was stirred for 3.5 hours at 0 ℃, concentrated under reduced pressure to afford a residue. The residue was dissolved with CH ₂Cl ₂ (5 mL) and extracted with water (2x5 mL) . The aqueous layer was freeze-dried and purified by Dowex-50wx Na ⁺ resin, using water as an elution solvent to give example 8 (42 mg, 77%) as a white powder.

¹H NMR (400 MHz, D ₂O) : δ 1.14 –1.26 (m, 2 H) , 1.58 –1.65 (m, 2 H) , 1.71 –1.82 (m, 2 H) , 1.90 –2.08 (m, 3 H) , 2.99 –3.16 (m, 3 H) , 3.38 (t, J = 11.4 Hz, 2 H) , 4.04 –4.13 (m, 1 H) , 3.82 –3.92 (m, 2 H) , 5.28 (s, 1 H) . LC-MS analysis: [M-Na] ^-= 361.2

Example 9

Sodium (2S, 5R) -7-oxo-2- (N- ( (R) -pyrrolidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (compound 81, Table 1)

Step 1: Synthesis of tert-butyl (3R) -3- ( (2S, 5R) -6- (benzyloxy) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) pyrrolidine-1-carboxylate (9_1)

To a solution of III (2.0 g, 3.9 mmol) in DCM (30 mL) at 0 ℃ was added tert-butyl (R) -3-aminopyrrolidine-1-carboxylate (2 mL, 5.9 mmol) and then added trimethyl aluminum (13.2 mL, 1 N in hexane, 13.2 mmol) dropwisely. The mixture was stirred at room temperature under argon overnight. Then 5.0 g silica was added under ice cooling bath, the mixture stirred for further 0.5 hour, 20 mL of methanol was added. Then the mixture was filtrated, washed with methanol and EtOAc, concentrated under reduced pressure to give a residue. The residue was dissolved with EtOAc, washed with saturated NH ₄Cl, dried by NaSO ₄, concentrated to provide the crude compound 9_1 (1.2 g) , which was directly used for next step without further purification. LC-MS analysis: [M+H] ⁺ = 444.1.

Step 2: Synthesis of tert-butyl (3R) -3- ( (2S, 5R, Z) -6- (benzyloxy) -N'- (tert-butoxycarbonyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) pyrrolidine-1-carboxylate (9_2)

A mixture of crude compound 9_1 (1.2 g obtained above) , (Boc) ₂O (2.0 g, 9.1 mmol) and TEA (1.3 mL, 10.0 mmol) in DCM (30 mL) was stirred at 20 ℃ overnight. The reaction mixture was concentrated under reduced pressure to give a residue. The residue was purified by flash chromatography on silica gel, eluting with 30%EtOAc in petroleum ether to provide the title compound 9_2 (1.5 g, 71%in two steps) as an oil.

¹H NMR (400 MHz, CDCl ₃) : δ 1.49 (s, 9 H) , 1.50 (s, 9 H) , 1.59 –1.69 (m, 2H) , 1.81 –1.93 (m, 2 H) , 2.05 –2.18 (m, 2 H) , 3.13 –3.41 (m, 4 H) , 3.49 –3.57 (m, 1 H) , 4.07 –4.15 (m, 1 H) , 4.17 –4.25 (m, 1 H) , 4.39 (d, J = 7.4 Hz, 1 H) , 4.92 (d, J = 11.3 Hz, 1 H) , 5.07 (d, J = 11.3 Hz, 1 H) , 5.69 –5.77 (m, 1 H) , 7.38 –7.46 (m, 5 H) . LC-MS analysis: [M+Na] ⁺ = 566.2.

Step 3: Synthesis of tert-butyl (3R) -3- ( (2S, 5R, Z) -N'- (tert-butoxycarbonyl) -6-hydroxy-7-oxo-1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) pyrrolidine-1-carboxylate (9_3)

10%Pd/C (wet, 55%water w/w, 0.3 g) was added to a solution of compound 9_2 (1.5 g, 2.76 mmol) in MeOH (20 mL) , and then stirred under H ₂ (balloon) at room temperature overnight, filtered through a pad of celite, rinsed with EtOAc. The filtrate was concentrated to give a residue, which was purified by flash chromatography on silica gel, eluting with 30%EtOAc in petroleum ether to provide the title compound 9_3 (0.34 g, 27%) .

¹H NMR (400 MHz, CDCl ₃) : δ 1.47 (s, 9 H) , 1.60 (s, 9 H) , 1.81 –1.91 (m, 3 H) , 2.02 –2.20 (m, 3 H) , 3.05 –3.23 (m, 2H) , 3.36 –3.49 (m, 2 H) , 3.57 –3.66 (m, 1 H) , 4.02 –4.15 (m, 2H) , 4.30 –4.39 (m, 1 H) , 6.02-6.11 (m, 1 H) .

Step 4: Synthesis of (3R) -3- ( (2S, 5R, Z) -N'- (tert-butoxycarbonyl) -7-oxo-6- (sulfooxy) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) pyrrolidine-1-carboxylate (9_4)

A mixture of compound 9_3 (0.11 g, 0.24 mmol) , SO ₃. NMe ₃ complex (0.10 g, 0.72 mmol) and TEA (0.1 mL, 0.72 mmol) in isopropyl alcohol and water (each 5 mL) was stirred at room temperature overnight. The reaction mixture was concentrated under reduced pressure to provide a residue, which was purified by prep. HPLC on a Agilent 10 prep-C18 250x21.2 mm column and lyophilized to give 9_4 (25 mg, 19%) as a white solid.

¹H NMR (400 MHz, DMSO-d ₆) : δ 1.45 (s, 9 H) , 1.47 (s, 9 H) , 1.76 –1.97 (m, 3 H) , 2.02-2.19 (m, 3 H) , 3.17 –3.26 (m, 1 H) , 3.33 –3.48 (m, 3 H) , 3.53 –3.62 (m, 1 H) , 3.92 –4.01 (m, 1 H) , 4.15 –4.30 (m, 2 H) , 5.25 (s, 1 H) , 7.10 (d, J = 6.5 Hz, 1 H) . LC-MS analysis: [M-H] ^-= 532.2.

Step 5: Synthesis of sodium (2S, 5R) -7-oxo-2- (N- ( (R) -pyrrolidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl sulfate (example 9)

TFA (0.5 mL) was added to a solution of compound 9_4 (50 mg, 0.09 mmol) in anhydrous CH ₂Cl ₂ (3 mL) at 0 ℃. The mixture was stirred for 4 hours at 0 ℃, and then concentrated under reduced pressure to give a residue. The residue was dissolved with CH ₂Cl ₂ (20 mL) and extracted with water (2x10 mL) . The aqueous layer was freeze-dried and purified by resin Dowex-50wx Na ⁺, using water as elution solvent to give example 9 (5 mg, 15%) as a white powder. LC-MS analysis: [M-Na] ^-= 332.1.

Pharmacological Methods

Antibacterial activity and synergistic activity:

Compounds of the present invention alone, meropenam (MER) alone, and as a combination with test antibiotic (meropenam) were tested for antimicrobial activity by determing minimum inhibitory concentration (MIC, mg/L) using the broth microdilution method according to the guidelines of the Clinical Laboratories and Stansards Insitute (“Methods for Dilution Antimicrobial Suscetibility Tests for Bacterial that Grow Aerobiclly” , Approved standard, 7th ed., Clinical and Laboratories Stanards Institute (CLSI) Document M7-A8, Wayne, Pa., USA, 2009) . Meropenam as a test antibiotic compound was dissolved in DMSO. Meropenam was then diluted in microbial growth medium (Mueller-Hinton Broth II, cation adjusted) resulting in a final concention range of 0.125-64 mg/L in serial two-fold dilution. In all cases the filnal DMSO concentraion was less than 0.5%. Bacteria were added to 96-well microtitre plates containing the serial two-fold dilutions of the compoumds; the final cell density was appoximately 5x10 ⁵ colony forming units/mL (CFU/mL) . Plates were incubated at 37 ℃ for 18-24 hours and read visually. The MIC, i.e. the lowest concentration of the test compound that inhibited visible growth of the bacteria, was recorded. The same assay conditions were used when the compounds of present invention alone, and as a combination with test meropenam antibiotic compound was tested for minmum inhibitory concentration (MIC, mg/L) . Whileest meropenam was serially diluted as descrribed above, a constant concentration of the present invention of 4 μg/mL was used. The test results listed in Table 2 table 3.

Bacterial strains that were used to evaluate the antimicrobial activity using the MIC determination included but were not limited to E. coli clinical isolate (strain 1) , E. coli 8739 (strain 2) , K. pneumoniae clinical isolate (strain 3) , K. pneumoniae 700603 (strain 4) , E. cloacae clinical isolate (strain 5) , E. cloacae 700323 (strain 6) , A. baumannii clinical isolate (strain 7) , A. baumannii 19606 (strain 8) , P. aeruginosa clinical isolate (strain 9) , P. aeruginosa 9027 (strain 10) .

Table 2: Antibacterial activity of example 1 to example 9 (Ex. 1 to Ex. 9, MIC, mg/L)

Table 3: Synergy of the inhibitor example 1 to example 9 (4 mg/L) in combination with meropenam (MER, MIC, mg/L)

Test for lactamase Inhibitory Activity:

The inhibitory activities of present compounds against various enzymes are measured by spectrophotometric assay using 490 nM and using nitrocefin as a substrate [J. Antimicrob. Chemother., 28, pp 775-776 (1991) ] . The concentration of inhibitor (IC ₅₀) which inhibits by 50%the reaction of hydrolysis of nitrocefin by the enzyme is determined.

In light of the data described herein, persons of skill in the art would expect that all of the compounds within the scope of formula (I) , salts of such compounds, solvates of such compounds, and salts thereof, and deuterated compounds of all such compounds, salts and solvates (i.e., compounds of formula (I) modified in that they have been deuterated, salts of compounds of formula (I) modified in that they have been deuterated, and solvates of compounds of formula (I) modified in that they have been deuterated) would be effective on their own as antibacterial compounds, and in combination with β-lactam antibiotics.

Efficacy of the β-lactamase inhibitors can be evaluated in combination with ceftazidime aztreonam, meropenem and other class of carbapenems and cephalosporins in murine infection models such as septicemia, pneumonia and thigh infection models (Ref: Andrea Endimiani et. al. Antimicrobial Agents and Chemotherapy, January 2011, page 82-85) . For murine acute lethal septicemia model, mice were infected by the intraperitoneal injection of the clinical strains resulting in death of the untreated controls within 24-48 hours. In particular, a fresh predetermined bacterial inoculum of approximately 3.3x10 ⁵ to 3.6x10 ⁵ CFU in 5% hog gastric mucin grown overnight. Thirty minutes post infection, a single subcutaneous dose of meropenam with and without β-lactamase inhibitor was initiated and the survival ratio monitored for 5 days twice daily. For each strain tested, the dosing regimen used are meropenam alone (doses of 512, 1024 &2048 mg/kg of body weight) and meropenam plus β-lactamase inhibitor at ratio of 2: 1, 4: 1, 8: 1, 16: 1 &32: 1 (meropenam doses were 4, 8, 16, 32 &64 mg/kg for each ratio) . The median effective dose for 50%protective dose (ED ₅₀) of animals was determined by a computerized program of Probit analysis. Survival rates stratified for different dosing regimen were also obtained. For experimental pneumoniae model, immunocompromised mice were used and intratracheally infected with Klebsiella pneumoniae strains. Mice in this model developed bacteraemia pneumoniae and fatal disease within 2 to 4 days with lung bacterial burden at 16-18 hours post infection of 10 ¹¹ to 10 ¹³ cfu/gm lung. Treatment with meropenam and inhibitor at a ratio of 2/1 &4/1 demonstrated efficacy with significant 3 to 6 log reduction in lung counts compared to meropenam alone and was relevant to the clinical situation. Human testing of the β-lactamase inhibitor can be conducted in combination with partner antibiotic at a set ratio utilizing standard clinical development practice.

Claims

A compound of formula (I) :

wherein:

M is hydrogen or a pharmaceutically acceptable salt forming cation,

Y = H or R,

When Y = R, R is a radical selected from any of the following groups:

(1) C _1-6 straight or branched chain alkyl which is optionally substituted;

(2) C _3-7 cycloalkyl which is optionally substituted;

(3) C _4-7 saturated heterocycles containing at least one heteroatom selected from O, N and S wherein the said heterocycle is optionally substituted, the ring S is optionally oxidized to S (O) or S (O) ₂ and the free ring N atom may optionally take a substituent.

(4) Heterocyclyl (C _1-6) alkyl wherein the said heterocycle has the same definition as defined in (3) ;

(5) C _5-7 membered heteroarylalkyl which is optionally substituted,

or a deuterated compound of any such compound.
The compound as recited in claim 1, wherein the radical R, when Y = R, is optionally substituted with one or two substituents independently selected from the following:

Lower alkyl, amino, substituted amino, alkoxy, hydroxyalkyl, halogen, hydroxy, carboxy, alkoxycarbonyl, haloalkyl, trifluoromethyl, trifluoromethyloxy, alkylamine, substituted alkylamine, carboxamide, thiocarboxamide, sulfonic acid, sulphate, acylamino, sulfonylamino, substituted or unsubstituted sulfonamide, substituted or unsubstituted urea, substituted or unsubstituted thiourea, oxyimino, hydroxamic acid, acyl, trifluoromethyl carbonyl, cyano, amidino, guanidino, aryloxy, heterocyclylalkyloxy, and heteroaryloxy.
The compound as recited in claim 1, which is selected from the following group of compounds:

and pharmaceutically acceptable salts of such compounds, and deuterated compounds of such compounds and salts.
The compound as defined in claim 1, wherein the compound is selected from the group consisting of:

(2S, 5R) -2-Carbamimidoyl-7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-Methylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-Ethylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-Isopropylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N-propylcarbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-Isobutylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-Neopentylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (Cyclopentylmethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (Cyclohexylmethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Acetamidoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Aminoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Hydroxyethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-oxo-2- (N- (2-Ureidoethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(Imino ( (2S, 5R) -7-oxo-6- (sulfooxy) -1, 6-diazabicyclo [3.2.1] octan-2-yl) methyl) glycine

(2S, 5R) -2- (N- (2-Methoxyethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (piperazin-1-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (tetrahydropyrimidin-1 (2H) -yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (piperidin-4-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (4-Methylpiperazin-1-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (3-Acetyltetrahydropyrimidin-1 (2H) -yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (1-Methylpiperidin-4-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Aminopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Amino-2-oxoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (piperidin-4-yloxy) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (pyrrolidin-3-yloxy) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (Azetidin-3-yloxy) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (piperidin-1-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (1-Acetylpiperidin-4-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (pyrrolidin-3-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (Azetidin-3-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (tetrahydro-2H-pyran-4-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (2- (pyrrolidin-1-yl) ethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (1, 1-Dioxidotetrahydro-2H-thiopyran-4-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Guanidinoethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (Guanidinooxy) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (1-Acetylpyrrolidin-3-yl) ethyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-cyclopropylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate

(2S, 5R) -2- (N-Cyclobutylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-Cyclopentylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3-Aminocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3- (Dimethylamino) cyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3- (Methylamino) cyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3-Acetamidocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Aminocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (Dimethylamino) cyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Acetamidocyclopentyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Acetamidocyclopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2- (Methylamino) cyclopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (4-Aminocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (4- (Dimethylamino) cyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N-Cycloheptylcarbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (4-Acetamidocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3-Aminocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3- (Dimethylamino) cyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3- (Methylamino) cyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (3-Acetamidocyclohexyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (2-Aminocyclopropyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (piperidin-4-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (1-Methylpiperidin-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (1-Acetylpiperidin-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

Ethyl 4- ( (2S, 5R) -7-oxo-6- (sulfooxy) -1, 6-diazabicyclo [3.2.1] octane-2-carboximidamido) piperidine-1-carboxylate;

(2S, 5R) -7-Oxo-2- (N- ( (R) -piperidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (R) -1-Methylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (R) -1-Acetylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- ( (S) -piperidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (S) -1-Methylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (S) -1-Acetylpiperidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- ( (S) -pyrrolidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (S) -1-Ethylpyrrolidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- ( (R) -pyrrolidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (R) -1-Formylpyrrolidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (R) -1-Acetylpyrrolidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (Azetidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (1-sulfamoylazetidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (1- ( (l2-Azanyl) (imino) methyl) azetidin-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (Azepan-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (tetrahydro-2H-thiopyran-4-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (tetrahydro-2H-pyran-4-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (1, 1-Dioxidotetrahydro-2H-thiopyran-4-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-oxo-2- (N- ( (R) -pyrrolidin-3-yl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- (1-Acetylazepan-3-yl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (pyrrolidin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (piperidin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (piperidin-4-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (piperidin-2-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (1, 3-Oxazinan-2-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (piperazin-2-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (1, 3-Thiazinan-2-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- ( (tetrahydrofuran-2-yl) methyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-oxo-2- (N- ( (tetrahydro-2H-pyran-4-yl) methyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate ;

(2S, 5R) -7-Oxo-2- (N- (pyridin-4-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (pyridin-3-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (pyrimidin-5-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -7-Oxo-2- (N- (pyridin-2-ylmethyl) carbamimidoyl) -1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (1 H-Imidazol-2-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

(2S, 5R) -2- (N- ( (1 H-Imidazol-1-yl) methyl) carbamimidoyl) -7-oxo-1, 6-diazabicyclo [3.2.1] octan-6-yl hydrogen sulfate;

or deuterated compounds of such compounds.
A method of treating a bacterial infection comprising administering to a mammal in need thereof an antibacterially effective amount of a compound as recited in anyone of claims 1-4.
A pharmaceutical composition containing, as an active ingredient, at least one compound as recited in anyone of claims 1-4.
A pharmaceutical composition containing, as an active ingredient, (i) at least one compound as recited in anyone of claims 1-4 and (ii) at least one β-lactam antibiotic, at least one salt of a β-lactam antibiotic, at least one hydrate of a β-lactam antibiotic or at least one prodrug of a β-lactam antibiotic.
The pharmaceutical composition of claim 7, wherein a ratio of the weight of (i) the compound of formula (I) to the weight of (ii) at least one β-lactam antibiotic, at least one salt of a β-lactam antibiotic, at least one hydrate of a β-lactam antibiotic or at least one prodrug of a β-lactam antibiotic, is in the range of 1: 30 to 30: 1.
A pharmaceutical composition containing, as an active ingredient, (i) at least one compound as recited in anyone of claims 1-4 and (ii) at least one antibiotic, at least one salt of an antibiotic, at least one hydrate of an antibiotic or at least one prodrug of an antibiotic.
The pharmaceutical composition of claim 9, wherein a ratio of the weight of (i) the compound of formula (I) to the weight of (ii) at least one antibiotic, at least one salt of an antibiotic, at least one hydrate of an antibiotic or at least one prodrug of an antibiotic, is in the range of 1: 30 to 30: 1.
The pharmaceutical composition of anyone of claims 7-10, further comprising a pharmaceutically acceptable carrier.
A method of treating a bacterial infection, comprising administering to a mammal in need thereof a combination of (i) an effective amount of a compound as recited in anyone of claims 1-4 and (ii) an effective amount of at least one β-lactam antibiotic, at least one salt of a β-lactam antibiotic, at least one hydrate of a β-lactam antibiotic or at least one prodrug of a β-lactam antibiotic.
The method of claim 12, wherein a ratio of the weight of (i) the compound of formula (I) to the weight of (ii) at least one β-lactam antibiotic, at least one salt of a β-lactam antibiotic, at least one hydrate of a β-lactam antibiotic or at least one prodrug of a β-lactam antibiotic, is in the range of 1: 30 to 30: 1.
A method of treating a bacterial infection, comprising administering to a mammal in need thereof a combination of (i) an effective amount of a compound as recited in anyone of claims 1-4 and (ii) an effective amount of at least one antibiotic, at least one salt of an antibiotic, at least one hydrate of an antibiotic or at least one prodrug of an antibiotic.
The method of claim 14, wherein a ratio of the weight of (i) the compound of formula (I) to the weight of (ii) at least one antibiotic, at least one salt of an antibiotic, at least one hydrate of an antibiotic or at least one prodrug of an antibiotic, is in the range of 1: 30 to 30: 1.
A molecular complex comprising a compound as recited in anyone of claims 1-4 and at least one solvent wherein the solvent comprises water.
Use of a compound as recited in anyone of claims 1-4 in preparation of a medicament for treating bacterial infections in mammals.
A process for preparing a compound recited in claim 1, wherein M = H, and wherein the process comprises:

step A, reacting substituted amine (NH ₂-Y) with the nitrile (III) in presence of reagent selected from the group consisting of trimethylaluminum, or triethylaluminum, or Lanthanum (III) or trifluoromethanesulfonate to provide an intermediate of formula (IV) , which could be protected in presence of reagent selected from the group consisting of Di-tert-butyl dicarbonate, trifluoroacetyl chloride and methyl trifluoroacetate to give an intermediate of formula (V) ;

step B, removing the benzyl protecting group of the intermediate (V) by hydrogenation to provide debenzylated product (VI) ;

step C, contacting compound (VI) with a sulfating reagent in the presence of a base to obtain compound of formula (I) after removal of protection group (PG) .

wherein Y is recited as in claim 1.