TW202116782A - Quinolone derivatives - Google Patents

Abstract

The invention relates to compounds of Formula I which are useful as antibacterial agents in medicine.

Description

Quinolinone derivatives

The present invention relates to novel antibacterial compounds, pharmaceutical compositions containing these compounds and the use of these compounds as medicaments, for example, in the treatment of bacterial infections. These compounds are suitable antimicrobial agents, which are effective against a variety of human and veterinary pathogens (including Gram-positive and Gram-negative aerobic and anaerobic bacteria) and especially It is effective against resistant strains of Pseudomonas aeruginosa and Enterobacteriacea (such as Klebsiella pneumonia).

The extensive use of antibiotics has led to a dramatic increase in microorganisms with gene-based resistance mechanisms. Modern medical and socioeconomic behaviors exacerbate the problem of drug resistance development by, for example, producing slow growth of pathogenic microorganisms in artificial joints and by supporting long-term host reservoirs, for example, in immunocompromised patients. In the hospital environment, more and more Staphylococcus aureus , Streptococcus pneumonia , Enterococcus spp., Enterobacteriaceae and Pseudomonas aeruginosa strains (main source of infection) Is becoming multi-drug resistant and therefore difficult to treat (if not impossible) -Staphylococcus aureus (S. aureus ) is resistant to β-lactam, quinolinone, and now even to vancomycin (Vancomycin) is resistant; Streptococcus pneumoniae ( S. pneumoniae ) is becoming resistant to penicillin (penicillin) or quinolinone antibiotics, and even resistant to new macrolides; Enterococcus ( Enteroccocci ) is resistant to quinolinone and vancomycin, and β-lactam antibiotics are not effective for these strains; enterobacteria are resistant to cephalosporin and quinolinone, and carbapenem is positive Loss of its efficacy (for example, Klebsiella pneumoniae resistant to carbapenem); P. aeruginosa is resistant to β-lactam and quinolinone. In addition, the incidence of multidrug-resistant Gram-negative strains (such as Enterobacteriaceae and Pseudomonas aeruginosa) is increasing, and newly emerged organisms (such as Acinetobacter spp.) that have been selected during treatment with currently used antibiotics ( Acinetobacter spp.) or Clostridium difficile (Clostridium difficile)) is becoming a practical problem in the hospital environment (SL Solomon et al., Antibiotic Resistance Threats In the United States: Stepping Back from the Brink, Academy of Family Physician , p. 940 pages, Volume 89, Volume 12, June 15, 2014). Therefore, there is a high medical need for new antibacterial agents with advantageous properties to overcome such multi-drug resistance in bacteria (especially Pseudomonas aeruginosa and enterobacteriaceae such as Klebsiella pneumoniae).

WO 2014/178008 and WO 2017/029602 disclose certain antibacterial compounds. The present invention provides novel antibacterial compounds with advantageous pharmacological properties.

式I 其中V 表示O或S(尤其O)；R¹ 表示C_1-4 烷基(尤其甲基或乙基)或C_3-5 環烷基(尤其環丙基)；且R² 表示氫或-(C=O)-O-CH₂ -O-(P=O)(OH)₂ ； 或其鹽(尤其醫藥學上可接受之鹽)。1) The first embodiment is about the compound of formula I

Formula I wherein V represents O or S (especially O); R ¹ represents C _1-4 alkyl (especially methyl or ethyl) or C _3-5 cycloalkyl (especially cyclopropyl); and R ² represents hydrogen Or -(C=O)-O-CH ₂ -O-(P=O)(OH) ₂ ; or a salt thereof (especially a pharmaceutically acceptable salt).

The definitions provided herein are intended to be uniformly applied to the compounds of formula I as defined in any one of Examples 1) to 9), and after necessary amendments to the details, in the entire specification and the scope of the patent application, unless expressly stated otherwise , Otherwise the definition provides a broader or narrower definition. It should be fully understood that the definitions or preferred definitions of terms are defined independently of (and combinations thereof) of any or all other terms as defined herein and can replace individual terms. If there is no other clear definition in the respective embodiments or the scope of the patent application, the groups defined herein are not substituted.

The term "alkyl" used alone or in combination refers to a saturated linear or branched hydrocarbon chain containing one to six carbon atoms. Examples are methyl, ethyl, n-propyl, isopropyl, n-butyl, tertiary butyl, second butyl, isobutyl, n-pentyl, 1,1-dimethylpropyl, 2, 2-Dimethylpropyl, 3-methylbutyl, 3-pentyl, 2-pentyl, 1,2-dimethylpropyl and 2-methylbutyl. _{The term "C xy} alkyl" used alone or in combination (X and Y are each an integer) refers to a saturated linear or branched hydrocarbon chain having x to y carbon atoms. _{Therefore, the term C 1-4} alkyl, alone or in combination with other groups, means a branched or straight chain group having one to four carbon atoms. Examples of C _1-4 alkyl groups are methyl, ethyl, n-propyl, isopropyl, n-butyl, tertiary butyl, sec-butyl and isobutyl; especially methyl or ethyl.

The term "cycloalkyl" used alone or in combination refers to a saturated monocyclic hydrocarbon ring containing three to six carbon atoms. The term "C _xy cycloalkyl" (X and Y are each an integer) refers to a saturated monocyclic hydrocarbon ring containing x to y carbon atoms. Examples of C _3-5 cycloalkyl groups are cyclopropyl, cyclobutyl and cyclopentyl; especially cyclopropyl.

2) Another example relates to the compound as in example 1), wherein V represents O.

3) Another embodiment relates to a compound as in any one of embodiments 1) or 2), wherein R ¹ represents C _1-4 alkyl (especially methyl or ethyl).

4) Another embodiment relates to a compound as in any one of embodiments 1) or 2), wherein R ¹ represents a C _3-5 cycloalkyl (especially cyclopropyl).

5) Another embodiment relates to a compound as in any one of embodiments 1) to 4), wherein R ² represents hydrogen.

6) Another embodiment relates to the compound according to any one of embodiments 1) to 4), wherein R ² represents -(C=O)-O-CH ₂ -O-(P=O)(OH) ₂ .

式II。7) Another embodiment relates to a compound as in any one of embodiments 1) to 6), which is also a compound of formula II (that is, the fragment 1,4-cyclohexylene in formula I has the same as in formula II Stereochemical name depicted)

Formula II.

8) Another example relates to the compound of example 1), which is selected from the group consisting of: 6-Fluoro-1-methyl-4-oxo-7-(4-{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][1 ,4]Thio-6-ylmethyl)-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]pyridine-3-carboxylic acid; 6-Fluoro-1-methyl-4-oxo-7-(4-{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][1 ,4]㗁𠯤-6-ylmethyl)-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]pyridine-3-carboxylic acid; 1-Ethyl-6-fluoro-4-oxo-7-(4{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][1, 4]Thio-6-ylmethyl)-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]pyridine-3-carboxylic acid; 1-Ethyl-6-fluoro-4-oxo-7-(4{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][1, 4]㗁𠯤-6-ylmethyl)-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]pyridine-3-carboxylic acid; 1-Cyclopropyl-6-fluoro-4-oxo-7-(4-{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][ 1,4]Thio-6-ylmethyl)-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]pyridine-3-carboxylic acid; 1-Cyclopropyl-6-fluoro-4-oxo-7-(4-{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][ 1,4]㗁𠯤-6-ylmethyl)-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]pyridine-3-carboxylic acid; 6-Fluoro-1-methyl-4-oxo-7-(4-{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][1 ,4]Thio-6-ylmethyl)-phosphinoyloxymethoxycarbonyl-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]㖠Pyridine-3-carboxylic acid; and 6-Fluoro-1-methyl-4-oxo-7-(4-{[(3-oxo-3,4-dihydro-2H-pyrido[2,3-b][1 ,4]㗁𠯤-6-ylmethyl)-phosphinooxymethoxycarbonyl-amino]-methyl}-cyclohexylamino)-1,4-dihydro-[1,8]㖠pyridine -3-carboxylic acid.

9) Another example relates to the compound of example 1), which is selected from the group consisting of: 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]Thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3- Formic acid 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3- Formic acid 1-Ethyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]Thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3- Formic acid 1-Ethyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3- Formic acid 1-cyclopropyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido [2,3-b][1,4]Thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3 -Formic acid; 1-cyclopropyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido [2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3 -Formic acid; 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]thio-6-yl)methyl)(((phosphinooxy)methoxy)carbonyl)amino)methyl)cyclohexyl)amino)-1, 4-dihydro-1,8-pyridine-3-carboxylic acid; and 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]㗁𠯤-6-yl)methyl)(((phosphinooxy)methoxy)carbonyl)amino)methyl)cyclohexyl)amino)-1, 4-dihydro-1,8-pyridine-3-carboxylic acid.

Based on the correlation of the different embodiments 1) to 7) as disclosed above, the following embodiments are therefore possible and anticipated, and are specifically disclosed in the following individualized forms: 2+1, 3+1, 3+2+1, 4+1, 4+2+1, 5+1, 5+2+1, 5+3+1, 5+3+2+1, 5+ 4+1, 5+4+2+1, 6+1, 6+2+1, 6+3+1, 6+3+2+1, 6+4+1, 6+4+2+1, 7+1, 7+2+1, 7+3+1, 7+3+2+1, 7+4+1, 7+4+2+1, 7+5+1, 7+5+2+ 1, 7+5+3+1, 7+5+3+2+1, 7+5+4+1, 7+5+4+2+1, 7+6+1, 7+6+2+ 1, 7+6+3+1, 7+6+3+2+1, 7+6+4+1 and 7+6+4+2+1.

In the above list, the number refers to the embodiment according to the number provided above, and the "+" indicates the relevance to another embodiment. The different individualization examples are separated by commas. In other words, for example, "3+2+1" means that the embodiment 3) depends on the embodiment 2) and depends on the embodiment 1), that is, the embodiment "3+2+1" corresponds to the further embodiment Example 1) of 2) and 3).

The compound of embodiment 1), wherein R ² represents -(C=O)-O-CH ₂ -O-(P=O)(OH) ₂ as defined above, in a biologically relevant environment (for example, in a removing a group - (C = O) -O- CH 2 -O- (P = O) (OH) phosphatase or any suitable presence of ₂ equivalents) exhibits antibacterial activity. In this environment, these compounds are usually converted to the compounds of Example 1), where R ² represents hydrogen. The compound of formula I, wherein R ² represents (C=O)-O-CH ₂ -O-(P=O)(OH) ₂ which is also known as a prodrug.

The compound of any one of Examples 1) to 9) is particularly active against bacteria and bacterial organisms. It can therefore be used in human and veterinary medicine to be particularly suitable for the prevention and treatment of local and systemic infections caused by such pathogens and diseases related to bacterial infections and chemotherapy. These bacterial infections include: and those caused by Streptococcus pneumoniae, catarrh Moraxella catarrhalis , Staphylococcus aureus, Staphylococcus epidermidis , Staphylococcus haemolyticus or Peptostreptococcus spp. infection-related pneumonia, otitis media, sinusitis, bronchitis , Tonsillitis and mastoiditis; pharyngitis and rheumatism related to infection by Streptococcus pyogenes , C and G streptococci, Corynebacterium diphtheria or Actinobacillus haemolyticum Fever and filamentous nephritis; Respiratory tract infections related to infections caused by Legionella pneumophila , Streptococcus pneumoniae or Chlamydia pneumonia ; Blood caused by Staphylococcus aureus and Staphylococcus hemolyticus Tissue infections, including endocarditis and osteomyelitis. The Staphylococcus aureus and Staphylococcus haemolyticus include strains resistant to known antibacterial agents such as but not limited to: β-lactam, vancomycin Aminoglycosides, quinolinones, chloramphenicol, tetracycline and macrolides; and Staphylococcus aureus, coagulase-negative staphylococci (ie, Staphylococcus epidermidis ( S. epidermidis ), Staphylococcus hemolyticus, etc.), Streptococcus pyogenes ( S. pyogenes ), Streptococcus agalactiae (Streptococcus agalactiae), Streptococcus group CF (Streptococcus microcolony), Streptococcus viridans (viridans streptococci), very small stick Corynebacterium minutissimum , Clostridium spp. , or Bartonella hensela infections related to uncomplicated skin and soft tissue infections, abscesses and postpartum fever; and those caused by Staphylococcus aureus or coagulase Uncomplicated acute urinary tract infections related to negative staphylococcal infections; urethritis and cervicitis; related to Chlamydia trachomatis and Haemophilus ducr eyi ), Treponema pallidum (Treponema pallidum), Ureaplasma urealyticum (Ureaplasma urealyticum), or Neiserria gonorrhea (Neiserria gonorrhea) infection related sexually transmitted diseases; related to Staphylococcus aureus (food poisoning and toxic shock syndrome) or A , B and C group streptococcal infections related toxin diseases; related to Chlamydia trachomatis ( C. trachomatis ), Neisseria gonorrhoeae ( N. gonorrhoeae ), Staphylococcus aureus, Streptococcus pneumoniae, Streptococcus pyogenes or Listeria spp. infection-related conjunctivitis, keratitis and dacryocystitis. The previously listed infections and pathogens are only construed as examples and by no means limiting.

The compound of one of Examples 1) to 9) can be used to prepare medicaments, and is suitable for the prevention or treatment of bacterial infections selected from the group consisting of respiratory tract infections, otitis media, meningitis, skin and soft tissue infections (regardless of concurrent Or uncomplicated), pneumonia (including hospital-acquired pneumonia), bacteremia, endocarditis, intra-abdominal infection, gastrointestinal infection, urinary tract infection, sexually transmitted infection, foreign body infection, osteomyelitis, Lyme Lyme disease, local infection or ophthalmological infection, and especially used to prevent or treat bacterial infections selected from the group consisting of: respiratory tract infection, otitis media, meningitis, skin and soft tissue infections (regardless of concurrent or uncomplicated ), pneumonia (including hospital-acquired pneumonia) and bacteremia.

The compound of one of Examples 1) to 9) can be further used for the preparation of medicaments, and is suitable for the treatment of infections mediated by: Gram-positive bacteria (such as Staphylococcus aureus, Bacillus cereus ) , Bacillus anthracis ( Bacillus anthracis ), Corynebacterium ( Corynebacterium spp. ) and Propionibacterium acnes (Propionibacterium acnes)), especially Gram-positive bacteria selected from the group consisting of: Cactus bacillus, Bacillus anthracis and Propionibacterium acnes. In particular, the compound of one of Examples 1) to 9) can be used to prepare medicaments, and is suitable for the treatment of bacterial infections mediated by Staphylococcus aureus (especially quinolone-resistant Staphylococcus aureus bacteria).

The compound of one of Examples 1) to 9) can be further used to prepare medicaments, and is suitable for the treatment of gram-negative bacteria (such as Escherichia coli, Klebsiella pneumoniae and other enterobacteria, green Infections mediated by Pseudomonas, Stenotrophomonas maltophilia, Neisseria meningitides , Moraxella catarrhalis and Bacteroides spp , Particularly selected from the group consisting of Gram-negative bacteria: Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Moraxella catarrhalis and Neisseria meningitidis. In particular, the compound of formula I and the pharmaceutically acceptable salt thereof in any one of Examples 1) to 9) can be used to prepare medicaments, and are suitable for the treatment of Klebsiella pneumoniae (especially multi-drug resistant). Or quinolinone-resistant Klebsiella pneumonia) and Pseudomonas aeruginosa-mediated bacterial infections.

One aspect of the present invention therefore relates to the use of a compound as one of Examples 1) to 9), which is used in the manufacture for the prevention or treatment of bacterial infections (in particular, the aforementioned Gram-negative One of bacteria-mediated infections or one of the aforementioned Gram-positive bacteria-mediated infections).

Another aspect of the present invention relates to the use of at least one of the compounds of Examples 1) to 9) as a medicament.

Yet another aspect of the present invention relates to a composition comprising at least one of the compounds of Examples 1) to 9) and further comprising at least one therapeutically inert excipient.

The pharmaceutical composition according to the present invention contains at least one of the compounds of Examples 1) to 9) as an active agent and optionally contains a carrier and/or diluent and/or adjuvant, and may also contain additional Know antibiotics.

Any method familiar to those who are familiar with this technology (see, for example, Remington, The Science and Practice of Pharmacy , 21st Edition (2005), Part 5, "Pharmaceutical Manufacturing" [published by Lippincott Williams & Wilkins]), by Combine the described compound of formula I or its pharmaceutically acceptable salt (as the case may be combined with other therapeutically valuable substances) with suitable non-toxic, inert, therapeutically compatible solid or liquid carrier materials and (depending on (Need) Common medical adjuvants are made into a galenical administration form to realize the preparation of pharmaceutical compositions.

Species such as pigs, ruminants, horses, dogs, cats, and poultry can be treated with a compound as in one of Examples 1) to 9).

Any reference herein to a compound such as one of Examples 1) to 9) should be understood to also refer to the salts of these compounds (and especially the pharmaceutically acceptable salts).

The compounds of Examples 1) to 9) can be used as medicaments, for example in the form of pharmaceutical compositions for enteral administration or especially parenteral administration (especially intravenous administration).

Another aspect of the present invention relates to a method for preventing or treating (preferably treating) a bacterial infection in a patient, which comprises administering to the patient a pharmaceutically active amount such as one of Examples 1) to 9) The compound of formula I or a pharmaceutically acceptable salt thereof. Therefore, the present invention provides a method for preventing or treating bacterial infections mediated by Gram-negative bacteria in patients (especially Klebsiella pneumoniae, and especially multidrug-resistant or quinolinone-resistant Klebsiella pneumoniae). Bacterial infections mediated by Pseudomonas and Pseudomonas aeruginosa), which comprises administering to the patient a pharmaceutically active amount of a compound of formula I or a pharmaceutically acceptable compound of one of Examples 1) to 9) salt. The present invention further provides a method for preventing or treating (preferably treating) bacterial infections mediated by Gram-positive bacteria in patients (especially Staphylococcus aureus, especially quinolinone-resistant Staphylococcus aureus). The method of bacterial infection), which comprises administering to the patient a pharmaceutically active amount of the compound of formula I or a pharmaceutically acceptable salt thereof as one of Examples 1) to 9).

The compounds of formula I according to the present invention can also be used for cleaning purposes, such as removing pathogenic microorganisms and bacteria from surgical instruments, catheters and artificial implants; making surfaces, rooms or areas sterile. For this purpose, the compound of formula I may be contained in solution, suspension/emulsion, spray, gel and/or dry powder formulations.

The compounds of Examples 1) to 9) can also be used in veterinary applications, such as the treatment of infections in livestock and companion animals. It can further constitute substances used to preserve inorganic and organic materials, especially all types of organic materials, such as polymers, lubricants, coatings, fibers, leather, paper, and wood.

The compound of any one of Examples 1) to 9) is suitable for use as active chemotherapeutic compounds in human and veterinary medicine and as substances for preserving inorganic and organic materials, especially all types of organic materials, such as polymers, Lubricants, coatings, fibers, leather, paper and wood.

The present invention also includes ^{compounds of formula I that are isotopically labeled, especially 2} H (deuterium), except that one or more atoms have been replaced by atoms having the same atomic number but different atomic masses from those commonly found in nature. , These compounds are the same as the compounds of formula I. The compounds of formula I and their salts which are labeled with isotopes, especially labeled with ² H (deuterium), are within the scope of the present invention. Substituting a heavier isotope ² H (deuterium) for hydrogen can produce greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, or can cause reduced inhibition of cytochrome P450 enzymes, resulting in, for example, an improved safety profile. In one embodiment of the present invention, the compound of formula I is not isotopically labeled, or it is only labeled with one or more deuterium atoms. In a sub-embodiment, the compound of formula I is not isotopically labeled at all. Isotopically-labeled compounds of formula I can be prepared in a similar manner to the methods described below, but using appropriate isotopic variants of suitable reagents or starting materials.

The term "pharmaceutically acceptable salt" refers to a salt that maintains the desired biological activity of the compound of the present invention and exhibits minimal undesirable toxicological effects. Depending on the presence of basic and/or acidic groups in the compounds of the present invention, such salts include inorganic acid or organic acid and/or base addition salts. For references, see, for example, "Handbook of Pharmaceutical Salts. Properties, Selection and Use.", P. Heinrich Stahl, Camille G. Wermuth (eds), Wiley-VCH, 2008 and "Pharmaceutical Salts and Co-crystals", Johan Wouters And Luc Quéré (eds), RSC Publishing, 2012.

The term "prevention" should be understood as equivalent to the term "prophylaxis".

In addition, the term "room temperature" as used herein refers to a temperature of about 25°C.

The compounds of formula I can be prepared according to the present invention using the procedures described below.

Compounds of formula I of the present description and examples the following abbreviations are used throughout:. Ac acetyl aq aqueous Boc-yl third butoxycarbonyl CHO Chinese hamster ovary CC column chromatography cPr cyclopropyl DAD diode array detector DCM two Methyl chloride DME 1,2 -Dimethoxyethane DMF N , N -Dimethylformamide DMAP 4-Dimethylaminopyridine DMPK Pharmacokinetics DMSO-d6 Deuterated dimethyl sulfide EA Ethyl acetate Ester ELSD Evaporative Light Scattering Detector ESI Electrospray Ionization Et Ethyl ex. Example Hept Heptane HPLC High Pressure Liquid Chromatography hv High Vacuum Conditions LC Liquid Chromatography LHS Left-hand side min min sec sec Me methyl MeCN Acetonitrile MeOH methanol MS mass spectrometry NMR nuclear magnetic resonance org. organic prep-HPLC preparative high pressure liquid chromatography rt room temperature sat. saturated SEM 2-trimethylsilyl-ethoxymethyl TBME tertiary butyl methyl ether TFA trifluoro Acetic acid THF Tetrahydrofuran TLC Thin layer chromatography t _R retention time (liquid chromatography) UV Ultraviolet light

All temperatures are stated in °C. Unless otherwise indicated, the reaction is carried out at rt.

Analytical method Use 0.2 mm disc: Merck, silicone 60 F _{254 to} perform analytical TLC characterization. Perform dissolution with EA, Hept, DCM, MeOH or a mixture thereof. Use UV or use a solution of KMnO ₄ (10 g), Na ₂ CO ₃ (20 g) and H ₂ O (1 L) for detection under subsequent heating; use Brunschwig 60A silicone (0.032-0.63 mm) or Use ISCO CombiFlash system and pre-filled SiO ₂ cartridge to perform column chromatography (CC) , use Hept-EA or DCM-MeOH mixture with appropriate gradient for dissolution; ¹ H-NMR (400 MHz, Bruker Avance 400 or 500 MHz, Bruker Avance 500 Cryoprobe) was used to characterize the compound. The chemical shift δ is given in ppm relative to the solvent used; multimodality: s = singlet, d = doublet, t = triplet, q = quartet, p = quintet, hex = hexat, hep = heptat, m = multiplet, br. = broad peak; coupling constant J is given in Hz; LC-MS is used to alternatively characterize compounds (Thermo Finnigan MSQPlus and Agilent G4220A). Use the following conditions to obtain analytical LC-MS data: LCMS1 data: column: Zorbax SB-Aq, 3.5 μm, 4.6 × 50 mm; injection volume: 1 μL; column oven temperature: 40°C; pump: Dionex HPG -3200RS; Replenishment pump: Dionex ISO-3100SD; DAD: Dionex DAD-30000RS; MS: Thermo MSQ Plus; ELSD: Sedere Sedex 85; Detection: UV 210 nm, ELSD and MS; MS ionization mode: ESI+; Eluent: A: H ₂ O + 0.04% TFA; and B: MeCN; flow rate: 4.5 mL/min; gradient: 5% B (0.00 min-0.01 min), 5% B to 95% B (0.01 min-1.00 min) , 95% B (1.00 min-1.45 min). The number of decimals given for the corresponding [M+H ⁺ ] peak value and residence time (t _R ) of each tested compound depends on the accuracy of the LC-MS device actually used.

Preparative HPLC equipment : HPLC pump: Gilson 333/334 or equivalent; autosampler: Gilson LH215 (with Gilson 845z ejector) or equivalent; degasser: Dionex SRD-3200 or equivalent; supplement Pump: Dionex ISO-3100A or equivalent; DAD detector: Dionex DAD-3000 or equivalent; MS detector: single quadrupole mass analyzer; Thermo Finnigan MSQ Plus or equivalent; MRA separator: MRA100 -000 splitter or equivalent; column: Zorbax SB-AQ 30 × 75 mm 5 μm; flow rate: 75 mL/min (for a column with a size of 30 × 75 mm); mobile phase: gradient mode A: water + 0.5% formic acid (acidic conditions); A: water + 0.5% ammonium hydroxide solution (25%) (alkaline conditions); B: acetonitrile.

Gradient : "Strong polarity" t[min] %A %B Flow rate mL/min 0.000 95 5 75 0.100 95 5 75 3.000 50 50 75 4.000 5 95 75 6.000 5 95 75 6.200 95 5 75 6.600 95 5 75 "Polar" t [min] %A %B Flow rate mL/min 0.000 90 10 75 0.010 90 10 75 4.000 5 95 75 6.000 5 95 75 6.200 90 10 75 6.600 90 10 75

General process of synthetic preparation :

Synthesis of BB1-1 of BB1: According to WO2011 / 037433 Preparation of 7-chloro-6-fluoro-1,4-dihydro-1,8-methyl-4-oxo-3-carboxylic acid㖠; BB1-2 : 7-chloro-1-ethyl-6-fluoro-4-oxo-1,4-dihydro-[1,8] pyridine-3-carboxylic acid is commercially available (CAS 79286 -73-0); BB1-3 : 7-chloro-1-cyclopropyl-6-fluoro-4-oxo-1,4-dihydro-[1,8]pyridine-3-carboxylic acid can be (CAS 100361-18-0) are commercially available; BB2 - (1r, 4r) -4- synthesis of hexan-1-amine hydrochloride (azidomethyl) cycloalkyl - the N - [(trans) - 4-(azidomethyl)cyclohexyl]-carbamic acid tert-butyl ester (305 mg; CAS 956352-36-6; prepared according to WO 2007/125952) for solution in dioxane (5 ml) Treated with 4M HCl in dioxane (1.2 mL) and further stirred at rt overnight. The obtained solid was collected by filtration and washed with anhydrous ether to obtain 212 mg (93% yield) of a colorless solid; MS1 (ESI, m/z): 196.40 [M+MeCN+H+]; t _R = 0.40 min.

BB3 Synthesis BB3-1 : 3- Pendant Oxygen -4-(2- Trimethylsilyl - Ethoxymethyl )-3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] Thio 𠯤 -6- formaldehyde

Step 1: 6-Chloro-4-(2-trimethylsilyl-ethoxymethyl)-4H-pyrido[2,3-b][1,4]thio-3-one contains Add 6-chloro-4H-pyrido[2,3-b][ to an ice-cold suspension of 60% sodium hydride dispersion in mineral oil (190 mg, 4.76 mmol, 1.2 equivalents) in DMF (10 mL) 1,4] Thiophene-3-one (800 mg, 3.97 mmol, 1 equivalent, CAS 2083653-83-0). The resulting light brown solution was stirred at 0°C for 15 min. Then (2-(chloromethoxy)ethyl)trimethylsilane (0.86 mL, 4.76 mmol, 1.2 equivalents) was added at 0°C, and the yellow solution was stirred at rt overnight. Water was added carefully, and the mixture was extracted three times with EA. The combined organic layer was washed with water, dried over MgSO ₄ , filtered and concentrated under reduced pressure. Then the crude yellow oil was purified by FC using CombiFlash (24 g SiO ₂ column, gradient: heptane:heptane/EA 3/1, within 18 min). The desired product (1.05 g, 80% yield) was obtained as a pale yellow oil. LCMS1 (ESI, m/z): t _R =1.09 min, 332.16 [M+H ⁺ ]; ¹ H NMR (500 MHz, DMSO-d6 δ: 8.39 (s, 1 H), 5.42 (s, 2 H) , 3.95 (s, 2 H), 3.58 (t, J = 7.8 Hz, 2 H), 0.83 (t, J = 7.9 Hz, 2 H), -0.10 (s, 9 H).

Step 2: 6-((E-styryl)-4-(2-trimethylsilyl-ethoxymethyl)-4H-pyrido[2,3-b][1,4]thiol 𠯤-3-ketone will contain 6-chloro-4-(2-trimethylsilyl-ethoxymethyl)-4H-pyrido[2,3-b][1,4]thia𠯤-3 -Ketone (985 mg, 2.97 mmol, 1 equivalent) and trans-2-phenylvinylboronic acid 97% (498 mg, 3.26 mmol, 1.1 equivalent) in anhydrous DMF (45 mL) and Na ₂ CO ₃ 2M (3 mL, 5.94 mmol, 2 equivalents) under N ₂ with [[1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (217 mg, 0.297 mmol, 0.1 equivalent) The resulting orange suspension was heated to 90°C and stirred at this temperature for 2 h. The mixture was cooled to rt, EA and water were added and the two layers were separated. The organic layer was washed 3× with water, washed with brine, MgSO _{4 was} dried, filtered and concentrated under reduced pressure. Then the _{crude product was purified by FC using CombiFlash (24 g SiO 2} column, gradient: heptane:heptane/EA 3/1 in 16 min). The desired product as a pale yellow oil (0.81 g, 68% yield). ¹ H NMR (500 MHz, DMSO-d6) δ: 8.35 (s, 1 H), 7.67-7.74 (m, 3 H), 7.34-7.45 (m, 4 H), 5.62 (s, 2 H), 3.94 (s, 2 H), 3.61 (t, J = 7.8 Hz, 2 H), 0.82-0.88 (m, 3 H),- 0.13 (s, 9 H). LCMS1 (ESI, m/z): t _R =1.19 min, 400.20 [M+H ⁺ ].

Step 3: 3-Pendant oxy-4-(2-trimethylsilyl-ethoxymethyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4 ] Thio 𠯤-6-formaldehyde. A solution of ELN173-4102.1 (810 mg, 2.03 mmol, 1 equivalent) in dioxane (15 mL) and water (2 ml) with 2,6-lutidine (0.708 mL, 6.08 mmol, 3 equivalents) , Treatment with 2.5 wt.% tertiary butanol (0.381 mL, 0.0304 mmol, 0.015 equivalent) containing osmium tetroxide solution and super pure (meta) sodium periodate (650 mg, 3.04 mmol, 1 equivalent). The yellow solution was stirred at rt over the weekend. Water (50 mL) was added and the mixture was extracted 3x with EA. The organic layers were combined, washed with brine, dried over MgSO _4, filtered and concentrated under reduced pressure. The crude product was purified by FC using CombiFlash (40 g SiO ₂ column, gradient: heptane:heptane/EA 1/1, within 20 min). The desired product (0.3 g, 47% yield) was obtained as an orange oil. ¹ H NMR (500 MHz, DMSO-d6) δ: 10.01 (s, 1 H), 8.71 (s, 1 H), 5.54 (s, 2 H), 4.05 (s, 2 H), 3.61 (t, J = 7.8 Hz, 2 H), 0.83 (t, J = 7.9 Hz, 2 H), -0.11 (s, 9 H). LCMS1 (ESI, m/z): t _R =1.0 min, 326.26 [M+H ⁺ ].

BB3-2 : 3- Pendant Oxygen -4-(2- Trimethylsilyl - Ethoxymethyl )-3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] 㗁 𠯤 -6- formaldehyde

Step 1: 6-Chloro-4-(2-trimethylsilyl-ethoxymethyl)-4H-pyrido[2,3-b][1,4]㗁𠯤-3-one will 2 -Chloro-N-(6-chloro-3-hydroxy-pyridine-2-yl)-acetamide (8100 mg, 32.8 mmol, 1 equivalent, CAS 2083653-89-6) in DMF (135 mL) The solution was treated with Cs ₂ CO ₃ (21420 mg, 65.7 mmol, 2.002 equivalents) once. Fever to 29°C. After 30 min, (2-(chloromethoxy)ethyl)trimethylsilane (7.12 mL, 39.4 mmol, 1.2 equivalents) was added dropwise. Slightly hot. Keep the temperature below 30°C with a water bath. After 20 min, the mixture was quenched on ice water and extracted with TBME. The aqueous phase was extracted again with TBME, and the organic phase was washed with water and brine, dried over MgSO ₄ and concentrated. The crude product was purified by SiO ₂ chromatography (Hept/EA 9:1, 4:1) to obtain the desired intermediate (8.54 g, 82% yield) as a colorless solid. LCMS1 (ESI, m/z): t _R =1.03 min, 316.27 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.01 (s, 1 H), 5.34 (s, 2 H), 5.06 (s, 2 H), 3.64 (m, 2 H), 0.88 (m, 3 H), -0.04 (s, 9 H).

Step 2: 4-(2-Trimethylsilyl-ethoxymethyl)-6-vinyl-4H-pyrido[2,3-b][1,4]㗁𠯤-3-ketone 6-Chloro-4-(2-Trimethylsilyl-ethoxymethyl)-4H-pyrido[2,3-b][1,4]㗁𠯤-3-one (853 mg, 2.7 mmol, 1 equivalent) and Si(triphenylphosphine)palladium(0) (156 mg, 0.135 mmol, 0.05 equivalent) were weighed in the flask and suspended in dioxane (10.8 mL). Tributyl(vinyl)tin 97% (1.63 mL, 5.4 mmol, 2 equivalents) was added and the mixture was degassed (vacuum/N ₂ , 5 times). The flask was heated to 100°C, sealed and stirred at this temperature for 4 h. The mixture was concentrated in vacuo and purified by FC (Hept/EA 9:1, 4:1) to obtain the desired intermediate (0.6 g, 76% yield) as a yellowish solid. LCMS1 (ESI, m/z): t _R =1.03 min, 308.28 [M+H ⁺ ]. ¹ H NMR (400 MHz, DMSO-d6) δ: 7.93 (s, 1 H), 6.81 (dd, J1 = 10.7 Hz, J2 = 17.2 Hz, 1 H), 6.18 (dd, J1 = 1.8 Hz, J2 = 17.2 Hz, 1 H), 5.45-5.48 (m, 3 H), 5.04 (s, 2 H), 3.66 (m, 2 H), 0.87 (m, 2 H), -0.07 (m, 9 H).

Step 3: 3-Pendant oxy-4-(2-trimethylsilyl-ethoxymethyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4 ]㗁𠯤-6-formaldehyde will 4-(2-trimethylsilyl-ethoxymethyl)-6-vinyl-4H-pyrido[2,3-b][1,4]㗁𠯤 A solution of -3-one (627 mg, 2.04 mmol, 1 equivalent) in 1,4-dioxane (25 mL) and water (5 mL) with 2.5 wt.% tertiary butanol containing osmium tetroxide solution (0.384 mL, 0.0306 mmol, 0.015 equivalents), (meta) sodium periodate (1309 mg, 6.12 mmol, 3 equivalents) and 2,6-lutidine (0.713 mL, 6.12 mmol, 3 equivalents) were treated with Stir at RT for 3 h. The solid was filtered and washed with EA. The filtrate was diluted with EA and washed with dilute HCl and brine, dried over MgSO ₄ and concentrated. The crude product was purified by SiO ₂ chromatography (Hept/EA 2:1) to obtain the title compound (0.48 g, 77% yield) as a colorless oil. LCMS1 (ESI, m/z): t _R =94 min, 310.21 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 9.94 (s, 1 H), 8.49 (s, 1 H), 5.45 (s, 2 H), 5.17 (s, 2 H), 3.68 (m, 2 H), 0.90 (m, 3 H), -0.03 (s, 9 H).

Coupling reaction LHS synthesis:

LHS1( R ¹ =Me): 7-(((1r,4r)-4-(aminomethyl)cyclohexyl)amino)-6-fluoro-1-methyl-4- pendant oxy-1,4 -Dihydro-1,8-pyridine-3-carboxylic acid will contain BB1-1 (6415 mg, 25 mmol, 1 equivalent) and BB2 (4767 mg, 25 mmol, 1 equivalent) of MeCN (150 mL) and DIPEA ( 15 mL, 87.5 mmol, 3.5 equivalents) of the mixture was heated to reflux for 6 h. The mixture was cooled to rt and poured onto water (300 mL). Adjust the pH to 4 by adding 2N HCl. The thick suspension was filtered and washed with water and TBME/MeOH 4:1 (100 ml). The wet coupling product was suspended in THF/H ₂ O 9:1 (120 mL) and triphenylphosphine (6557 mg, 25 mmol, 1 equivalent) was added. The mixture was heated at 60°C for 24 h. The mixture was cooled to rt, filtered, and the residue was washed with TBME and dried under high vacuum. The desired building block was obtained as a beige solid (7.6 g, after 2 steps, 87% yield). LCMS1 (ESI, m/z): t _R =0.58 min, 349.28 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.88 (s, 1 H), 8.13-8.19 (m, 1 H), 7.93 (d, J = 10.7 Hz, 1 H), 3.93-4.01 (m, 4 H), 2.42 (d, J = 6.3 Hz, 2 H), 2.00-2.02 (m, 2 H), 1.85 (m, 2 H), 1.42 (m, 2 H), 1.19-1.25 (m, 1 H), 0.98-1.06 (m, 2 H).

LHS2 ( R ¹ =Et): 7-(((1r,4r)-4-(aminomethyl)cyclohexyl)amino)-1-ethyl-6-fluoro-4-oxo-1,4 -Dihydro-1,8-Petidine-3-carboxylic acid is similar to LHS1, starting from BB1-2 and BB2 to prepare the title compound (70% yield, colorless solid). ¹ H NMR (500 MHz, DMSO-d6) δ: 8.91 (s, 1 H), 8.20 (d, J = 7.2 Hz, 1 H), 7.94 (d, J = 10.7 Hz, 1 H), 4.48 (q , J = 7.0 Hz, 2 H), 3.91-3.97 (m, 1 H), 2.41 (d, J = 6.4 Hz, 2 H), 1.98-2.01 (m, 2 H), 1.85-1.87 (m, 2 H), 1.36-1.46 (m, 5 H), 1.18-1.25 (m, 1 H), 0.96-1.04 (m, 2 H). LCMS1 (ESI, m/z): t _R =0.6 min, 363.13 [M+H ⁺ ].

LHS3 ( R ¹ =cPr): 7 -(((1r,4r)-4-(aminomethyl)cyclohexyl)amino)-1-cyclopropyl-6-fluoro-4-oxo-1, 4-Dihydro-1,8-Pinidine-3-carboxylic acid is similar to LHS1, starting from BB1-3 and BB2 to prepare the title compound (85% yield, colorless solid). LCMS1 (ESI, m/z): t _R =0.58 min, 375.17 [M+H ⁺ ].

Instance 1 : 6- fluorine -1- methyl -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] Thio 𠯤 -6- base ) methyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

1.1. 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4 -((((3-side oxy-4-((2-(trimethylsilyl (Yl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]thio-6-yl)methyl)amino)methyl) Cyclohexyl) amino)-1,4-dihydro-1,8-pyridine-3-carboxylic acid LHS1 (2975 mg, 8.54 mmol, 1 equivalent) and BB3-1 (2863 mg, 8.8 mmol, 1.03 equivalent) The solution in AcOH (0.488 mL, 8.54 mmol, 1 equivalent) and DCM/MeOH 4:1 (50 mL) was stirred at rt for 30 min. NaBH(OAc) ₃ (4505 mg, 21.3 mmol, 2.5 equivalents) was added and the mixture was stirred at rt for another 30 min. The reaction was quenched by adding water and DCM. Stir for 15 min at rt. The phases were separated and the aqueous phase was basified _{by adding NaHCO 3 solution.} Extract again with DCM. The organic phase was washed with dilute NaHCO ₃ solution, dried over MgSO ₄ and concentrated. The crude product was purified by chromatography on SiO ₂ (2.5-20% MeOH in DCM) to give the title intermediate (3.8 g, 68% yield) as an off-white foam. LCMS1 (ESI, m/z): t _R =0.88 min, 658.43 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.89 (s, 1 H), 8.27 (s, 1 H), 8.18 (d, J = 7.1 Hz, 1 H), 7.94 (d, J = 10.7 Hz , 1 H), 5.51 (s, 2 H), 3.96-4.04 (m, 1 H), 3.93 (s, 3 H), 3.89 (s, 2 H), 3.80 (s, 2 H), 3.56 (m , 2 H), 2.42 (d, J = 6.5 Hz, 2 H), 2.00 (m, 2 H), 1.90 (m, 2 H), 1.39-1.47 (m, 3 H), 1.02-1.10 (m, 2 H), 0.80 (m, 2 H), -0.12-0.10 (m, 9 H).

1.2. 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyridine And [2,3-b][1,4]thio (thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine- A solution of 3-formic acid example 1.1 (3800 mg, 5.78 mmol, 1 equivalent) in DCM (55 mL) was treated with TFA (4.42 mL, 57.8 mmol, 10 equivalents) and stirred at rt for 6 h. Complete the conversion. The mixture was concentrated in vacuo and azeotroped once with toluene. The residue was crystallized from MeOH (55 ml), and the mixture was basified by adding ammonia 25% (4 equivalents). The resulting fine suspension was stirred at rt for 45 min, filtered, washed with a small amount of MeOH, and dried under high vacuum. The title compound (3.1 g, 100%) was obtained as a yellow solid. LCMS1 (ESI, m/z): t _R =0.66 min, 528.33 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 11.07-11.39 (br, 1 H), 8.89 (m, 1 H), 8.20 (m, 2 H), 7.94 (d, J = 10.6 Hz, 1 H ), 3.96-4.02 (m, 1 H), 3.93 (s, 3 H), 3.79 (m, 4 H), 1.88-2.02 (m, 4 H), 1.43 (m, 3 H), 1.06 (m, 2 H).

Instance 2 : 6- fluorine -1- methyl -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] 㗁 𠯤 -6- base ) methyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

2.1. 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-4-((2-(trimethylsilyl (Yl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl) Cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3-carboxylic acid started with LHS1 and BB3-2 and followed the procedure of Example 1.1 to obtain the title compound as an off-white foam (77% yield ). LCMS1 (ESI, m/z): t _R =0.6 min, 642.29 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO) δ: 8.89 (m, 1 H), 8.18 (d, J = 7.3 Hz, 1 H), 7.92-7.95 (m, 2 H), 5.40-5.42 (m, 2 H ), 5.01-5.03 (m, 2 H), 3.98 (m, 1 H), 3.93 (s, 3 H), 3.75 (s, 2 H), 3.64 (m, 2 H), 2.43 (d, J = 6.5 Hz, 2 H), 2.00-2.02 (m, 2 H), 1.88-1.91 (m, 4 H), 1.40-1.47 (m, 3 H), 1.02-1.10 (m, 2 H), 0.86 (m , 2 H), -0.06 (m, 9 H).

2.2. 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyridine And [2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine- Starting with example 2.1 and following the procedure of example 1.2, the title compound (64%) was obtained as an off-white solid. LCMS1 (ESI, m/z): t _R =0.61 min, 512.33 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.89 (s, 1 H), 8.17 (d, J = 7.1 Hz, 1 H), 7.93-7.95 (m, 1 H), 7.79-7.81 (m, 1 H), 4.87 (s, 2 H), 3.95-4.03 (m, 1 H), 3.91-3.93 (m, 3 H), 3.69 (m, 2 H), 2.41 (d, J = 6.5 Hz, 2 H), 1.99-2.02 (m, 2 H), 1.89 (m, 2 H), 1.38-1.47 (m, 3 H), 1.02-1.11 (m, 2 H).

Instance 3 : 1- Ethyl -6- fluorine -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] Thio 𠯤 -6- base ) methyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

3.1. 1-Ethyl-6-fluoro-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-4-((2-(trimethylsilyl (Yl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]thio-6-yl)methyl)amino)methyl) Cyclohexyl) amino)-1,4-dihydro-1,8-pyridine-3-carboxylic acid started with LHS2 and BB3-1 and followed the procedure of Example 1.1 to obtain the title compound (80% yield) as an off-white foam rate). LCMS1 (ESI, m/z): t _R =0.91 min, 672.43 [M+H ⁺ ].

3.2. 1-Ethyl-6-fluoro-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyridine And [2,3-b][1,4]thio (thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine- Starting with Example 3.1 and following the procedure of Example 1.2, 3-formic acid was purified by preparative HPLC (column: Zorbax, acidic conditions, polar gradient, as specified in the analytical method) to obtain the title compound as an off-white solid (42%). LCMS1 (ESI, m/z): t _R =0.69 min, 541.99 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.92 (s, 1 H), 8.19 (d, J = 8.2 Hz, 3 H), 7.95 (d, J = 10.7 Hz, 1 H), 4.49 (q , J = 7.0 Hz, 2 H), 3.87-4.00 (m, 1 H), 3.78 (s, 2 H), 3.75 (s, 2 H), 2.42 (d, J = 6.5 Hz, 2 H), 1.99 -2.01 (m, 2 H), 1.89-1.91 (m, 2 H), 1.38-1.48 (m, 6 H), 1.01-1.09 (m, 2 H).

Instance 4 : 1- Ethyl -6- fluorine -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] 㗁 𠯤 -6- base ) methyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

4.1. 1-Ethyl-6-fluoro-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-4-((2-(trimethylsilyl (Yl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl) Cyclohexyl) amino)-1,4-dihydro-1,8-pyridine-3-carboxylic acid started with LHS2 and BB3-2 and followed the procedure of Example 1.1 to obtain the title compound (95% yield) as an off-white foam rate). LCMS1 (ESI, m/z): t _R =0.88 min, 656.45 [M+H ⁺ ].

4.2. 1-Ethyl-6-fluoro-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyridine And [2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine- Starting with Example 4.1 and following the procedure of Example 1.2, 3-formic acid was purified by preparative HPLC (column: Zorbax, acidic conditions, polar gradient, as specified in the analytical method) to obtain the title compound as an off-white solid (43%). LCMS1 (ESI, m/z): t _R =0.66 min, 526.30 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.91 (s, 1 H), 8.19 (d, J = 6.8 Hz, 2 H), 7.94 (m, 1 H), 7.83 (s, 1 H), 5.76 (s, 1 H), 4.88 (s, 2 H), 4.48 (q, J = 6.9 Hz, 2 H), 3.95 (m, 1 H), 3.75 (s, 2 H), 2.44-2.48 (m , 2 H), 1.99-2.04 (m, 2 H), 1.89 (d, J = 11.8 Hz, 2 H), 1.43-1.47 (m, 3 H), 1.39 (t, J = 7.0 Hz, 3 H) , 1.02-1.09 (m, 2 H).

Instance 5 : 1- Cyclopropyl -6- fluorine -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] Thio 𠯤 -6- base ) methyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

5.1. 1-Cyclopropyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-4-((2-(trimethyl Silyl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]thio-6-yl)methyl)amino)methyl )Cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3-carboxylic acid started with LHS3 and BB3-1 and followed the procedure of Example 1.1 to obtain the title compound (92%) as an off-white foam Yield). LCMS1 (ESI, m/z): t _R =0.92 min, 684.45 [M+H ⁺ ].

5.2. 1-Cyclopropyl-6-fluoro-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyridine 𠯤[2,3-b][1,4]thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine -3-carboxylic acid started from Example 5.1 and followed the procedure of Example 1.2. After purification by preparative HPLC (column: Zorbax, acidic conditions, polar gradient, as specified in the analytical method), the title was obtained as an off-white solid Compound (34%). LCMS1 (ESI, m/z): t _R =0.70 min, 554.28 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.57 (s, 1 H), 8.19 (m, 3 H), 7.94 (d, J = 10.6 Hz, 1 H), 4.02 (m, 1 H), 3.78 (s, 2 H), 3.75 (s, 2 H), 3.66-3.70 (m, 1 H), 2.43 (d, J = 6.5 Hz, 2 H), 2.05-2.08 (m, 2 H), 1.90 (d, J = 11.6 Hz, 2 H), 1.39-1.46 (m, 3 H), 1.02-1.18 (m, 6 H).

Instance 6 : 1- Cyclopropyl -6- fluorine -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] 㗁 𠯤 -6- base ) methyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

6.1. 1-Cyclopropyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-4-((2-(trimethyl Silyl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl )Cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3-carboxylic acid starting with LHS3 and BB3-2 and following the procedure of Example 1.1, the title compound (85% Yield). LCMS1 (ESI, m/z): t _R =0.88 min, 668.44 [M+H ⁺ ].

6.2. 1-Cyclopropyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyridine 𠯤[2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine -3-carboxylic acid started from Example 6.1 and followed the procedure of Example 1.2. After purification by preparative HPLC (column: Zorbax, acidic conditions, polar gradient, as specified in the analytical method), the title was obtained as an off-white solid Compound (56%). LCMS1 (ESI, m/z): t _R =0.66 min, 538.32 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 8.57 (s, 1 H), 8.19 (m, 2 H), 7.93 (d, J = 10.6 Hz, 1 H), 7.82 (s, 1 H), 4.87 (s, 2 H), 4.02 (m, 1 H), 3.73 (s, 2 H), 3.68 (m, 1 H), 2.46 (d, J = 6.6 Hz, 2 H), 1.99-2.10 (m , 2 H), 1.88-1.91 (m, 2 H), 1.39-1.46 (m, 3 H), 1.13-1.18 (m, 2 H), 1.01-1.12 (m, 4 H).

Instance 7 : 6- fluorine -1- methyl -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] Thio 𠯤 -6- base ) methyl )((( Phosphonoxy ) Methoxy ) Carbonyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

7.1. 7-(((1r,4r)-4-((((chloromethoxy)carbonyl)((3-side oxy-4-((2-(trimethylsilyl)ethoxy) (Methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]thio-6-yl)methyl)amino)methyl)cyclohexyl)amino) -6-Fluoro-1-methyl-4-oxo-1,4-dihydro-1,8-pyridine-3-carboxylic acid in Example 1.1 (250 mg, 0.38 mmol, 1 equivalent) in DCM (4 Add chloromethyl chloroformate (0.0372 mL, 0.418 mmol, 1.1 equivalents) to a yellow solution in saturated NaHCO _{3 (4 mL).} The resulting solution (two layers) was stirred vigorously for 2 h at rt. The mixture was diluted with DCM and water. The phases were separated and the aqueous layer was extracted again with DCM. The combined organic layer was washed with brine, dried over MgSO ₄ and concentrated under reduced pressure to give the title intermediate (272 mg, 95% yield) as a yellow foam. LCMS1 (ESI, m/z): t _R =1.19 min, 750.29 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 15.65-15.65 (m, 1 H), 11.26-11.29 (m, 1 H), 8.89 (d, J = 1.4 Hz, 1 H), 8.19 (m, 1 H), 8.06-8.08 (m, 1 H), 7.92-7.95 (m, 1 H), 5.46 (m, 2 H), 4.51 (m, 2 H), 3.97-4.05 (m, 1 H), 3.94 (d, J = 2.3 Hz, 3 H), 3.80 (d, J = 2.7 Hz, 2 H), 3.22-3.26 (m, 2 H), 2.00-2.02 (m, 2 H), 1.61-1.79 ( m, 3 H), 1.37-1.42 (m, 2 H), 1.08-1.16 (m, 2 H).

7.2. 7-(((1r,4r)-4-(((((((di-third-butoxyphosphoryl)oxy)methoxy)carbonyl)((3-oxo-4 -((2-(Trimethylsilyl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]thio-6-yl )Methyl)amino)methyl)cyclohexyl)amino)-6-fluoro-1-methyl-4-oxo-1,4-dihydro-1,8-pyridine-3-carboxylic acid Example 7.1 A solution of (270 mg, 0.36 mmol, 1 equivalent) in DME (5 mL) was treated with tetra-n-butylammonium di-tert-butyl phosphate (199 mg, 0.432 mmol, 1.2 equivalents) and heated at 55°C 4 h. The mixture was cooled to rt and partitioned between EA and water. The organic layer was washed with water and brine, dried over MgSO ₄ and concentrated under reduced pressure to obtain the desired intermediate (322 mg, 97% yield) as a yellow foam, which was used without purification. In one step. LCMS1 (ESI, m/z): t _R = 1.22 min, 924.75 [M+H ⁺ ].

7.3. 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyridine And [2,3-b][1,4]thio-6-yl)methyl)(((phosphinooxy)methoxy)carbonyl)amino)methyl)cyclohexyl)amino)- 1,4-Dihydro-1,8-pyridine-3-carboxylic acid A solution of Example 7.2 (320 mg, 0.346 mmol, 1 equivalent) in DCM (5 mL) was cooled to 0°C and TFA (0.663 mL, 8.66 mmol, 25 equivalents). Then the yellow solution was stirred at 0°C for 2 h. TBME (7 mL) was added to the solution, and the resulting suspension was stirred at 0°C for 5 min, filtered and the solid was dried under high vacuum for 5 min. The resulting yellow solid was suspended in MeOH (5 mL), and 25% NH ₄ OH (0.333 mL, 8.66 mmol, 25 equivalents) was added. The resulting mixture was then stirred at rt for 30 min. The suspension was then concentrated under reduced pressure (water bath at rt). The residue was diluted with water/MeCN and acidified with formic acid, and the resulting solution was purified by preparative HPLC using acidic conditions (Zorbax column; absolute polarity gradient) and the collected samples were lyophilized. The colorless lyophilisate was wet-milled in TBME (20 mL), the suspension was filtered, and the solid was dried under high vacuum to obtain the title compound (112 mg, 47% yield) as a colorless solid. LCMS1 (ESI, m/z): t _R =0.70 min, 682.29 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 15.65-15.65 (m, 1 H), 11.26-11.29 (m, 1 H), 8.89 (d, J = 1.4 Hz, 1 H), 8.19 (m, 1 H), 8.06-8.08 (m, 1 H), 7.92-7.95 (m, 1 H), 5.46 (m, 2 H), 4.51 (m, 2 H), 3.97-4.05 (m, 1 H), 3.94 (d, J = 2.3 Hz, 3 H), 3.80 (d, J = 2.7 Hz, 2 H), 3.22-3.26 (m, 2 H), 2.00-2.02 (m, 2 H), 1.61-1.79 ( m, 3 H), 1.37-1.42 (m, 2 H), 1.08-1.16 (m, 2 H).

Instance 8 : 6- fluorine -1- methyl -4- Pendant Oxygen -7-(((1r,4r)-4-(((((3- Pendant Oxygen -3,4- Dihydro -2H- Pyridine 𠯤 and [2,3-b][1,4] 㗁 𠯤 -6- base ) methyl )((( Phosphonoxy ) Methoxy ) Carbonyl ) Amino ) methyl ) Cyclohexyl ) Amino )-1,4- Dihydro -1,8- Pyridine -3- Formic acid

8.1. 7-(((1r,4r)-4-((((chloromethoxy)carbonyl)((3-side oxy-4-((2-(trimethylsilyl)ethoxy) (Methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino) -6-Fluoro-1-methyl-4-oxo-1,4-dihydro-1,8-pyridine-3-carboxylic acid in Example 2.1 (250 mg, 0.39 mmol, 1 equivalent) in DCM (4 Add chloromethyl chloroformate (0.0381 mL, 0.428 mmol, 1.1 equivalents) to the yellow solution in _{saturated NaHCO 3} (4 mL) and saturated NaHCO 3 (4 mL). The resulting solution (two layers) was stirred vigorously for 2 h at rt. The mixture was diluted with DCM and water. The phases were separated and the aqueous layer was extracted again with DCM. The combined organic layer was washed with brine, dried over MgSO ₄ and concentrated under reduced pressure to obtain the title intermediate (285 mg, 100% yield) as an off-white foam, which was used without further purification In the next step. LCMS1 (ESI, m/z): t _R =1.16 min, 734.23 [M+H ⁺ ].

8.2. 7-(((1r,4r)-4-((((((di-third-butoxyphosphoryl)oxy)methoxy)carbonyl)((3-side oxy-4 -((2-(Trimethylsilyl)ethoxy)methyl)-3,4-dihydro-2H-pyrido[2,3-b][1,4]㗁𠯤-6-yl )Methyl)amino)methyl)cyclohexyl)oxy)-6-fluoro-1-methyl-4-oxo-1,4-dihydro-1,8-pyridine-3-carboxylic acid Example 8.1 A solution of (280 mg, 0.381 mmol, 1 equivalent) in DME (5 mL) was treated with tetra-n-butylammonium di-tert-butyl phosphate (211 mg, 0.458 mmol, 1.2 equivalents) and heated at 55°C 5 h. The mixture was cooled to rt and partitioned between EA and water. The organic layer was washed with water and brine, dried over MgSO ₄ and concentrated under reduced pressure to obtain the desired intermediate (345 mg, 100% yield) as an off-white foam, which was used without further purification In the final step. LCMS1 (ESI, m/z): t _R =1.19 min, 908.69 [M+H ⁺ ].

8.3. 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyridine And [2,3-b][1,4]㗁𠯤-6-yl)methyl)(((phosphinooxy)methoxy)carbonyl)amino)methyl)cyclohexyl)amino)- 1,4-Dihydro-1,8-pyridine-3-carboxylic acid A solution of Example 8.2 (340 mg, 0.374 mmol, 1 equivalent) in DCM (5 mL) was cooled to 0°C and TFA (0.717 mL, 9.36 mmol, 25 equivalents) treatment. Then the yellow solution was stirred at 0°C for 3 h. TBME (7 mL) was added to the solution and the resulting suspension was stirred at 0°C for 5 min, filtered and the solid was dried under high vacuum for 5 min. The resulting solid was suspended in MeOH (5 mL) and 25% NH ₄ OH (0.361 mL, 9.36 mmol, 25 equivalents) was added. The resulting mixture was stirred at rt for 20 min, and then concentrated under reduced pressure. The residue was diluted with water/MeCN and acidified with formic acid, and the resulting solution was purified by preparative HPLC using acidic conditions (Zorbax column; absolute polar gradient, collected at 266 nm) and the collected samples were lyophilized. The colorless lyophilisate was wet-milled in TBME (20 mL), the suspension was filtered, and the solid was dried under high vacuum to obtain the desired product (144 mg, 58% yield) as a yellowish solid. LCMS1 (ESI, m/z): t _R =0.67 min, 666.40 [M+H ⁺ ]. ¹ H NMR (500 MHz, DMSO-d6) δ: 15.63-15.67 (m, 1 H), 11.62 (s, 1 H), 8.89 (s, 1 H), 8.18 (d, J = 7.5 Hz, 1 H ), 7.95 (d, J = 10.7 Hz, 1 H), 7.72 (d, J = 4.4 Hz, 1 H), 5.45-5.51 (m, 2 H), 4.88 (s, 2 H), 4.47 (m, 2 H), 3.91-4.04 (m, 4 H), 1.99-2.02 (m, 2 H), 1.64-1.77 (m, 3 H), 1.35-1.45 (m, 2 H), 1.07-1.17 (m, 2 H).

Pharmacological properties The minimum inhibitory concentration of bacterial growth: the minimum inhibitory concentration (MIC; mg/L) is measured in the cation-adjusted Mueller-Hinton medium by the micro-dilution method, which follows the " Methods for Dilution Antimicrobial Susceptibility Tests" for Bacteria that Grow Aerobically ", approved standard. 7th edition, description given in Clinical and Laboratory Standards Institute (CLSI) document M7-A7, Wayne, PA, USA (2006). The compounds (inv.) according to Examples 1 to 6 of the present invention were tested against several Gram-positive and Gram-negative bacteria. Staphylococcus aureus A798 is a multi-drug resistant strain (specifically resistant to quinolinone), Klebsiella pneumoniae A651 and Acinetobacter baumannii T6474 are both multi-drug resistant strains (specifically resistant to quinolinone) , And E. coli ATCC25922 and Moraxella catarrhalis A894 are quinolinone sensitive strains. The corresponding anti-bacterial test results are given in Table 1 below (MIC, in mg/L). Table 1 Instance number MIC of Staphylococcus aureus A798 MIC of Escherichia coli ATCC 25922 MIC of Moraxella catarrhalis A894 MIC of Klebsiella pneumoniae A651 MIC of Acinetobacter baumannii T6474 1 (inv.) ≤0.063 ≤0.063 ≤0.063 0.125 ≤0.063 2 (inv.) 2 0.25 ≤0.063 2 1 3 (inv.) 0.125 0.063 ≤0.031 0.5 ≤0.031 4 (inv.) 4 0.5 0.25 4 1 5 (inv.) 0.125 0.125 ≤0.031 1 0.063 6 (inv.) 4 0.5 0.25 4 0.5

The example of the present invention was tested against wild-type E. coli A-1261 in the presence of alkaline phosphatase (from bovine intestinal mucosa: Sigma P6774-2KU; batch: SLBB1168) and in the absence of alkaline phosphatase Compounds of 7 and 8 (inv.). The corresponding antibacterial test results are given in Table 2 below (MIC, in mg/L). Table 2 Instance number Active metabolite instance number MIC of Escherichia coli A-1261 In the absence of alkaline phosphatase In the presence of alkaline phosphatase (2 iU/mL) 7 (inv.) 1 (inv.) 1 ≤0.031 8 (inv.) 2 (inv.) 16 0.25

In mammalian cell analysis (CHO cells), a genetic toxicology test for compounds to assess their ability to cause DNA damage is performed in vitro. The formation of extranuclear micronuclei and smaller DNA staining particles was studied in mononuclear and binuclear cells treated with increasing concentrations of test compounds for 24 h. The cells were fixed, the cytoplasm was stained with CellmaskRed, and the DNA was stained with Hoechst 33342 (Mutat Res. 630:1-13, 2007). A high-content screening microscope (Opera Phenix, Perkin Elmer) was used to read the microtiter plate, and the manufacturer’s Harmony software was used to score cells, nuclei, and micronuclei using an optimized algorithm.

Cytotoxicity is determined by calculating the quotient of the number of effective cells in the compound-treated sample and the number of effective cells in only the DMSO-treated sample and plotting the number as the fraction of effective cells relative to the compound concentration. Cells are detected based on their cytoplasmic staining, and if the cell contains 1 to 2 intact regular nuclei, it is defined as effective. List the micronuclei associated with effective cells, and if the score of effective cells after baseline correction is ≥0.4, the induction of micronucleus formation is tabulated as the ratio of untreated cells. Based on the data obtained with known antibiotics, the threshold of significant increase is set to 2 times.

The genotoxic potential of the compounds (inv.) and reference compounds (ref.) according to Examples 1 to 6 of the present invention were measured in the micronucleus (MN) analysis, and the corresponding test results are given in Table 3 below. All reference compounds were prepared as described in WO 2014/178008. Table 3 Instance number Example number, as disclosed in WO 2014/178008 The highest concentration (µM), at which no significant increase in MN formation is observed 1 (inv.) 150 2 (inv.) 600 3 (inv.) 150 4 (inv.) 300 5 (inv.) 75 6 (inv.) ＞300 1 (ref.) 18.75 8 (ref.) 37.5 13 (ref.) 75 14 (ref.) 150 15 (ref.) 37.5

By detecting nuclear integrity, the cytotoxic potential of the compounds (inv.) and reference compounds (ref.) according to Examples 1 to 6 of the present invention were measured in parallel with micronucleus analysis, and the corresponding test results (IC ₅₀ ) µM meter) is given in Table 4 below. All reference compounds were prepared as described in WO 2014/178008. Table 4 Instance number Example number, as disclosed in WO 2014/178008 Cytotoxicity IC ₅₀ (µM) 1 (inv.) 217 2 (inv.) ＞1200 3 (inv.) 174 4 (inv.) 948 5 (inv.) 126 6 (inv.) ＞300 1 (ref.) 44 8 (ref.) 109 13 (ref.) 107 14 (ref.) 300 15 (ref.) 74

The compounds according to the present invention not only exhibit excellent antimicrobial activity against several bacterial strains (Table 1), they also exhibit superior pharmacological properties (Tables 3 and 4). The compounds according to Examples 7 and 8 exhibited antimicrobial activity in a biologically relevant environment by forming the compounds of Examples 1 and 2 (Table 2).

Claims (15)

A compound of formula I

Formula I wherein V represents O or S; R ¹ represents C _1-4 alkyl or C _3-5 cycloalkyl; and R ² represents hydrogen or -(C=O)-O-CH ₂ -O-(P= O) (OH) ₂ ; or a pharmaceutically acceptable salt thereof. Such as the compound of claim 1, wherein V represents O. The compound according to any one of claim 1 or 2, wherein R ¹ represents a C _1-4 alkyl group. The compound according to any one of claims 1 to 2, wherein R ¹ represents a C _3-5 cycloalkyl group. The compound according to any one of claims 1 to 4, wherein R ² represents hydrogen. The compound of any one of claims 1 to 4, wherein R ² represents -(C=O)-O-CH ₂ -O-(P=O)(OH) ₂ . As the compound of any one of claims 1 to 6, it is also a compound of formula II.

Formula II Such as the compound of claim 1, which is selected from the group consisting of: 6-Fluoro-1-methyl-4-oxo(oxo)-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyridine 𠯤[2,3-b][1,4]thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine -3-carboxylic acid; 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3- Formic acid 1-Ethyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]Thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3- Formic acid 1-Ethyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3- Formic acid 1-cyclopropyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido [2,3-b][1,4]Thio-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3 -Formic acid; 1-cyclopropyl-6-fluoro-4-oxo-7-(((1r,4r)-4-((((3-oxo-3,4-dihydro-2H-pyrido [2,3-b][1,4]㗁𠯤-6-yl)methyl)amino)methyl)cyclohexyl)amino)-1,4-dihydro-1,8-pyridine-3 -Formic acid; 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]thio-6-yl)methyl)(((phosphinooxy)methoxy)carbonyl)amino)methyl)cyclohexyl)amino)-1, 4-dihydro-1,8-pyridine-3-carboxylic acid; and 6-Fluoro-1-methyl-4-side oxy-7-(((1r,4r)-4-((((3-side oxy-3,4-dihydro-2H-pyrido[ 2,3-b][1,4]㗁𠯤-6-yl)methyl)(((phosphinooxy)methoxy)carbonyl)amino)methyl)cyclohexyl)amino)-1, 4-dihydro-1,8-pyridine-3-carboxylic acid; Or its pharmaceutically acceptable salt. A composition comprising at least one compound according to any one of claims 1 to 8, and further comprising at least one pharmaceutically acceptable excipient. The compound of any one of claims 1 to 8, which is used as a medicament. The compound according to any one of claims 1 to 8, which is used to prevent or treat bacterial infections. The compound according to any one of claims 1 to 8, which is used to prevent or treat bacterial infections mediated by Gram-negative bacteria. Such as the compound of any one of claims 1 to 8, which is used to prevent or treat bacterial infections mediated by Staphylococcus aureus , Klebsiella pneumoniae , Escherichia coli ( Escherichia coli ), Acinetobacter baumannii , Moraxella catarrhalis and/or Pseudomonas aeruginosa . A method for treating bacterial infections, the method comprising administering a compound according to any one of claims 1 to 8 to an individual in need. A use of the compound according to any one of claims 1 to 8 in the manufacture of a medicament for the prevention or treatment of bacterial infections.