CEPHALOSPORIN ANTIBACTERIAL COMPOUND AND PREPARATION METHOD THEREFOR

Abstract

The present disclosure relates to a cephalosporin antibacterial compound and a preparation method therefor. The cephalosporin antibacterial compound can exhibit antibacterial activity against Gram-negative bacteria and other bacteria.

Claims

1. A compound of formula I or a pharmaceutically acceptable salt thereof, ##STR00036## wherein, X is N, CH or CCl; T is S, S?O, CH.sub.2 or O; E is ##STR00037## wherein R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, halogen, phenyl, alkylthio, and alkyl optionally substituted with carbamoyl; R.sub.11 and R.sub.12 are each independently selected from the group consisting of hydrogen, carboxyl, and alkyl optionally substituted with carbamoyl; m is an integer of 1-5; F is a single bond; A is C.sub.1-C.sub.6 alkylene, C.sub.2-C.sub.6 alkenylene or C.sub.2-C.sub.6 alkynylene; each R.sub.5 is independently selected from the group consisting of halogen, hydroxy, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 haloalkyl and C.sub.1-C.sub.6 haloalkoxy; G.sub.1 is ##STR00038## R.sub.1 and R.sub.2 are each independently selected from the group consisting of hydrogen, halogen, hydroxy, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 haloalkyl and C.sub.1-C.sub.6 haloalkoxy; R.sub.3 is C.sub.1-C.sub.6 alkyl substituted with ##STR00039## R.sub.3 is selected from the group consisting of hydrogen, hydroxy, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, wherein the alkyl and alkoxy are optionally substituted with one or more substituents selected from the group consisting of C.sub.1-C.sub.6 alkyl, halogen, hydroxy, sulfhydryl, NR.sub.iR.sub.j, ##STR00040## oxo, thio, C(O)R.sub.k, C(O)OR.sub.k, C(S)R.sub.k, nitro, cyano, C.sub.1-C.sub.6 alkoxy and C.sub.1-C.sub.6 alkylthioether group; each R.sub.m is independently selected from the group consisting of hydrogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 alkoxy, hydroxy, 6- to 10-membered aryl and 5- to 8-membered heteroaryl, wherein the alkyl, alkoxy, aryl and heteroaryl are optionally substituted with one or more substituents selected from the group consisting of C.sub.1-C.sub.6 alkyl, halogen, hydroxy, sulfhydryl, amino, carboxyl, nitro, cyano and C.sub.1-C.sub.6 alkoxy; each R.sub.4 is independently selected from the group consisting of halogen, hydroxy, sulfhydryl and NR.sub.iR.sub.j; R.sub.i and R.sub.j are each independently selected from the group consisting of hydrogen, hydroxy, C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; each R.sub.k is independently selected from the group consisting of hydrogen, C.sub.1-C.sub.6 alkyl, C.sub.1-C.sub.6 haloalkyl, hydroxy, C.sub.1-C.sub.6 alkoxy and NR.sub.iR.sub.j, wherein the alkyl, haloalkyl and alkoxy are optionally substituted with one or more substituents selected from the group consisting of C.sub.1-C.sub.6 alkyl, halogen, hydroxy, sulfhydryl, NR.sub.iR.sub.j, oxo, thio, carboxyl, nitro, cyano, C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkylthioether group, 3- to 6-membered cycloalkyl, 3- to 6-membered heterocyclyl, 6- to 10-membered aryl and 5- to 8-membered heteroaryl; p is an integer of 0-5; q is an integer of 0-5; x is an integer of 3-8; n is an integer of 0-3.

2. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein A is C.sub.1-C.sub.6 alkylene.

3. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.1 and R.sub.2 are both hydrogen atoms.

4. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein x is an integer of 3-6.

5. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.3 is selected from C.sub.3-C.sub.6 alkyl substituted with ##STR00041##

6. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein ##STR00042## is ##STR00043## wherein each R.sub.n is independently selected from the group consisting of C.sub.1-C.sub.6 alkyl, hydroxy and halogen, and r is independently an integer of 0-5.

7. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sub.3 is selected from the group consisting of hydrogen and C.sub.1-C.sub.6 alkyl substituted with ##STR00044##

8. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein ##STR00045## is ##STR00046## wherein each R.sub.n is independently selected from the group consisting of C.sub.1-C.sub.6 alkyl, hydroxy and halogen, and r is independently an integer of 0-5.

9. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein the compound of formula I is selected from ##STR00047##

10. The compound or the pharmaceutically acceptable salt thereof according to claim 1, being selected from ##STR00048## or a pharmaceutically acceptable salt thereof.

11. An isotopically substituted form of the compound or the pharmaceutically acceptable salt thereof according to claim 1.

12. A pharmaceutical composition comprising the compound or the pharmaceutically acceptable salt thereof according to claim 1 and a pharmaceutically acceptable carrier, diluent or excipient.

13. A method of treating or preventing a disease caused by gram-negative bacteria in a subject in need thereof, the method comprising administering to the subject the-he compound or the pharmaceutically acceptable salt thereof according to claim 1.

14. A method of treating or preventing a disease caused by pathogenic bacteria in a subject in need thereof, the method comprising administering to the subject the compound or the pharmaceutically acceptable salt thereof according to claim 1.

15. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein ##STR00049## is ##STR00050##

16. The compound or the pharmaceutically acceptable salt thereof according to claim 1, wherein ##STR00051## is ##STR00052##

17. The isotopically substituted form of the compound or the pharmaceutically acceptable salt thereof according to claim 11, wherein the isotopic substitution is a substitution with a deuterium atom.

18. The method according to claim 13, wherein the disease is selected from the group consisting of airway infectious diseases, urinary system infectious diseases, respiratory system infectious diseases, septicemia, nephritis, cholecystitis, oral infectious diseases, endocarditis, pneumonia, bone marrow membrane myelitis, otitis media, enteritis, empyema, traumatic infectious diseases and opportunistic infections.

19. The method according to claim 13, wherein the gram-negative bacteria is selected from the group consisting of E. coli, Klebsiella, Serratia, Enterobacter, Citrobacter, Morganella, Providencia, Proteus, Haemophilus, Moraxella, Pseudomonas aeruginosa, Pseudomonas other than P. aeruginosa, Stenotrophomonas, Burkholderia and Acinetobacter.

Description

DETAILED DESCRIPTION

[0117] The preparation of the compound described herein and a pharmaceutically acceptable salt thereof is further described below in conjunction with examples, which are not intended to limit the scope of the present disclosure.

[0118] Experimental procedures without conditions specified in the examples of the present disclosure are generally conducted according to conventional conditions, or according to conditions recommended by the manufacturers of the starting materials or commercial products. Reagents without specific origins indicated are commercially available conventional reagents.

[0119] The structures of the compounds were determined by nuclear magnetic resonance (NMR) spectroscopy and/or mass spectrometry (LCMS). NMR shifts (?) were given in 10.sup.?6 (ppm). NMR analysis was performed on a Bruker AVANCE-400 nuclear magnetic resonance instrument, with deuterated dimethyl sulfoxide (DMSO-d.sub.6), deuterated chloroform (CDCl.sub.3) and deuterated methanol (CD.sub.3OD) as solvents and tetramethylsilane (TMS) as an internal standard. The spatial configurations of the optical isomers (isomers) of the compounds can be further confirmed by determining single crystal parameters.

[0120] HPLC analysis was performed on Waters ACQUITY ultra high performance LC, Shimadzu LC-20A systems, Shimadzu LC-2010HT series, or Agilent 1200 LC high performance liquid chromatograph (ACQUITY UPLC BEH C18 1.7 ?m 2.1?50 mm column, Ultimate XB-C18 3.0?150 mm column, or Xtimate C18 2.1?30 mm column).

[0121] MS analysis was performed on a Waters SQD2 mass spectrometer in the positive/negative ion scan mode with a mass scan range of 100-1200.

[0122] Chiral HPLC analysis was performed using a Chiralpak IC-3 100?4.6 mm I.D., 3 ?m; Chiralpak AD-3 150?4.6 mm I.D., 3 ?m; Chiralpak AD-3 50?4.6 mm I.D., 3 ?m; Chiralpak AS-3 150?4.6 mm I.D., 3 ?m; Chiralpak AS-3 100?4.6 mm I.D., 3 ?m; ChiralCel OD-3 150?4.6 mm I.D., 3 ?m; Chiralcel OD-3 100?4.6 mm I.D., 3 ?m; ChiralCel OJ-H 150?4.6 mm I.D., 5 ?m; or Chiralcel OJ-3 150?4.6 mm I.D., 3 ?m column. Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates, 0.15-0.2 mm layer thickness, were adopted for thin-layer chromatography (TLC) analysis and 0.4-0.5 mm layer thickness for TLC separation and purification.

[0123] Flash column purification was performed on a Combiflash Rf150 (TELEDYNE ISCO) or Isolara one (Biotage) system.

[0124] Normal phase column chromatography generally used 100-200 mesh, 200-300 mesh or 300-400 mesh Yantai Huanghai silica gel as a carrier, or used a Changzhou Santai pre-fill ultrapure normal phase silica gel column (40-63 ?m, 60 g, 12 g, 25 g, 40 g, 80 g or other specifications).

[0125] Reversed-phase column chromatography generally used a Changzhou Santai pre-fill ultrapure C18 silica gel column (20-45 ?m, 100 ?, 40 g, 80 g, 120 g, 220 g or other specifications).

[0126] High pressure column purification was performed on a Waters AutoP system in combination with a Waters XBridge BEH C18 OBD Prep Column, 130 ?, 5 ?m, 19 mm?150 mm or Atlantis T3 OBD Prep Column, 100 ?, 5 ?m, 19 mm?150 mm. Preparative chiral chromatography used a DAICEL CHIRALPAK IC (250 mm?30 mm, 10 ?m) or Phenomenex-Amylose-1 (250 mm?30 mm, 5 ?m) column.

Known starting materials in the present disclosure may be synthesized using or according to methods known in the art, or may be purchased from Shanghai Titan Scientific, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Darui Chemicals, and other companies.

[0127] In the example, all reactions can be performed under nitrogen atmosphere unless otherwise specified.

[0128] The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of argon or nitrogen.

[0129] The hydrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of hydrogen.

[0130] Pressurized hydrogenation reactions were performed using a Parr 3916EKX hydrogenator and a Qinglan QL-500 hydrogenator, or an HC2-SS hydrogenator.

[0131] Hydrogenation reactions generally involve 3 cycles of vacuumization and hydrogen purging.

[0132] Microwave reactions were performed on a CEM Discover-S 908860 microwave reactor.

[0133] In the examples, a solution refers to an aqueous solution unless otherwise specified.

[0134] In the examples, the reaction temperature refers to room temperature, i.e., 20? C. to 30? C., unless otherwise specified.

[0135] The reaction progress in the examples is monitored by thin-layer chromatography (TLC). The developing solvent for reactions, the eluent system for column chromatography purification and the developing solvent system for thin-layer chromatography include: A: dichloromethane/methanol system, B: n-hexane/ethyl acetate system, C: petroleum ether/ethyl acetate system, D: petroleum ether/ethyl acetate/methanol system, and E: petroleum ether/tetrahydrofuran system. The volume ratio of the solvents is adjusted according to the polarity of the compound, or by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

Example 1

[0136] ##STR00030## ##STR00031## ##STR00032##

Step 1

[0137] To a solution of compound 1-1 (synthesized according to the method of the patent CN106661052A, 1.5 g, 3.50 mmol) in DMF (35 mL) were added DMTMM (1.55 g, 5.26 mmol) and DIEA (905 mg, 7.0 mmol). After the reaction was purged with argon three times, 1-A (525 mg, 7.0 mmol) was added to the above reaction mixture, and the reaction was then stirred at room temperature for 2 h. The reaction mixture was slowly added to water, and a solid product gradually precipitated. The solid product obtained by filtration was dissolved in DCM. The solution was extracted, and the organic phase was separated, washed with water and then with saturated brine, dried over anhydrous sodium sulfate, filtered and evaporated to dryness to give the product 1-2 (1.6 g, yield: 94%).

[0138] LC/MS (ESI): m/z 486.1 [M+H].sup.+

Step 2

[0139] A solution of oxalyl chloride (1.05 g, 8.24 mmol) in DCM (10 mL) was cooled to ?78? C. in a dry ice/acetone bath, and DMSO (1.29 g, 16.5 mmol) was added dropwise under argon atmosphere. The reaction was then stirred. A solution of 1-2 (1.6 g, 3.30 mmol) in DCM (6 mL) was added slowly dropwise to the above reaction mixture, and the mixture was stirred at ?78? C. Et.sub.3N (3.34 g, 33.0 mmol) was added, and the mixture was stirred at ?78? C. The reaction was then slowly warmed to 0? C. and was completed. The reaction mixture was quenched with H.sub.2O and then extracted with DCM. The organic phase was washed with water and then with saturated brine, dried over anhydrous sodium sulfate, filtered and evaporated to dryness to give a crude product. The crude product was purified by flash chromatography on silica gel to give the product 1-3 (1.28 g, yield: 80%).

[0140] LC/MS (ESI): m/z 484.0 [M+H].sup.+

Step 3

[0141] To a solution of substrate 1-3 (1.28 g, 2.65 mmol) in DCM (26 mL) was added 1-B (605 mg, 5.30 mmol). After the reaction mixture was cooled in an ice-water bath, NaBH(OAc).sub.3 (1.68 g, 7.95 mmol) was added. The reaction was warmed to room temperature and stirred. After the reaction was complete, the reaction mixture was quenched with NaHCO.sub.3 solution and then extracted with DCM. The organic phase was washed with water and then with saturated brine, dried over anhydrous sodium sulfate, filtered and evaporated to dryness to give a crude product. The crude product was purified by flash chromatography on silica gel to give the product 1-4 (735 mg, yield: 48%).

[0142] LC/MS (ESI): m/z 582.1 [M+H].sup.+

Step 4

[0143] To a solution of 1-1 (650 mg, 1.51 mmol) in DMF (13 mL) were added HATU (961 mg, 2.52 mmol) and DIEA (490 mg, 3.78 mmol). The reaction was stirred at room temperature under argon atmosphere. 1-4 (735 mg, 1.26 mmol) was added to the above reaction mixture, and the reaction was then stirred at room temperature. The reaction mixture was slowly added to water, filtered and dried in vacuo to give the product 1-5 (1.2 g, yield: 96%).

[0144] LC/MS (ESI): m/z 992.1 [M+H].sup.+

Step 5

[0145] Compound 1-5 (546 mg, 0.55 mmol), compound 1-6 (synthesized according to the method of the patent WO2016035847, 398 mg, 0.5 mmol) and sodium iodide (225 mg, 1.5 mmol) were added to N,N-dimethylformamide (1.5 mL), and the reaction was stirred. After the reaction was complete, the reaction flask was cooled in an ice-water bath. PBr.sub.3 (360 mg, 1.33 mmol) was added, and the reaction was stirred. After the reaction was complete, the reaction mixture was added dropwise to aqueous NaHSO.sub.3 solution and filtered, and the filter cake was washed with clear water and dried in vacuo to give crude 1-7 (1.13 g).

[0146] LC/MS (ESI): m/z 1737.1 [M].sup.+

Step 6

[0147] Compound 1-7 (1.13 g, 0.5 mmol) was added to anisole (2 mL) and trifluoroacetic acid (8 mL), and the mixture was allowed to react at room temperature. Methyl tent-butyl ether was added, and the mixture was stirred and then filtered. The filter cake was rinsed with MTBE and then dried to give a crude product (600 mg). The crude product was purified by preparative HPLC to give compound 1 (4 mg).

[0148] HRMS: 979.1874 [M+H].sup.+

[0149] .sup.1H-NMR (400 MHz, DMSO-d.sub.6) ?: 1.44 (s, 3H), 1.46 (s, 3H), 1.66-2.06 (m, 6H), 2.93-3.88 (m, 14H), 3.04-3.05 (m, 2H), 4.00-4.03 (m, 2H), 4.82-5.13 (m, 1H), 5.18 (d, 1H), 5.74 (dd, 1H), 6.54 (d, 1H), 6.60 (d, 1H), 6.73-6.85 (m, 3H), 7.28 (s, 2H), 8.05-8.33 (m, 1H), 9.46 (br s, 2H), 10.26 (br s, 2H).

Example 2

[0150] ##STR00033##

Step 1

[0151] To a reaction flask were added 2-1 (30.3 g, 174.1 mmol, 1.0 eq), 2,4-dimethoxybenzaldehyde (29.5 g, 177.6 mmol, 1.02 eq), methanol (300 mL) and anhydrous sodium sulfate (24.7 g, 174.1 mmol, 1.0 eq), and the reaction was stirred at room temperature. The reaction mixture was then cooled in an ice-water bath, and sodium borohydride (3.3 g, 87.0 mmol, 0.5 eq.) was added in portions. The reaction was stirred for 5 min and then stirred at room temperature until it was complete. Acetic acid (3.3 mL) was added, and the mixture was stirred, filtered and washed with ethyl acetate. The filtrate was concentrated, and the residue was mixed with water and ethyl acetate and stirred. The aqueous phase was separated and extracted with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give the product 2-2 (58.0 g, yield: 102.8%).

[0152] MS m/z 325.1 [M+H].sup.+

Step 2

[0153] 2-2 (2.0 g, 6.2 mmol, 1.0 eq), S-propylene oxide (0.54 g, 9.3 mmol, 1.5 eq) and EtOH (20 mL) were added to a reaction flask, and the reaction was heated to 60? C. LC-MS monitoring showed that the starting material was consumed completely. The reaction mixture was concentrated, and the crude product was purified by column chromatography to give the product 2-3 (1.2 g, yield: 51.1%).

[0154] MS m/z 383.2 [M+H].sup.+

Step 3

[0155] 2-3 (1.2 g, 3.2 mmol, 1.0 eq), DCM (20 mL) and TEA (0.65 g, 6.4 mmol, 2.0 eq) were added to a reaction flask and cooled in an ice-water bath. MsCl (0.54 g, 4.7 mmol, 1.5 eq) was slowly added dropwise, and the reaction was stirred. LC-MS monitoring showed that the starting material was consumed completely. The reaction mixture was washed with water and saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated to give crude 2-4 (1.34 g). The crude product was directly used in the next step.

[0156] MS m/z 461.1 [M+H].sup.+

Step 4

[0157] The crude 2-4 (1.34 g, 3.2 mmol, 1.0 eq), tetrahydropyrrole (0.56 g, 8.0 mmol, 2.5 eq), MeCN (15 mL) and potassium carbonate (0.66 g, 4.8 mmol, 1.5 eq) were added to a reaction flask, and the reaction was heated to 40? C. and stirred. LC-MS monitoring showed that the starting material was consumed completely. The reaction mixture was mixed with water (30 mL) and EA (30 mL) and stirred. The aqueous phase was separated and washed with EA. The organic phases were combined, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate and concentrated. The crude product was purified by column chromatography to give 2-5 (0.66 g, yield over two steps: 48.5%).

[0158] MS m/z 436.1 [M+H].sup.+

Step 5

[0159] 2-5 (600 mg, 1.38 mmol) and TFA (10 mL) were added to a reaction flask, and the compound was dissolved. The reaction was heated to 60? C. and completed. The reaction mixture was concentrated to dryness and triturated with MTBE (10 mL). The supernatant was removed, and the resulting oil 2-6 was concentrated in vacuo to remove most of the solvent and directly used in the next step.

[0160] MS m/z 186.1 [M+H].sup.+

Step 6

[0161] The crude 2-6 and 1-1 (1.31 g, 3.05 mmol) were dissolved in DCM (15 mL), and the solution was stirred at 0? C. DIPEA (2.02 mL, 12.22 mmol) and HATU (1.39 g, 3.67 mmol) were added. The reaction mixture was naturally warmed to room temperature and was allowed to react until the reaction was complete. Water (10 mL) was added, and the organic phase was separated, washed with water, dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The crude product was separated by column chromatography (DCM:MeOH=10:1) to give 2-7 (0.85 g).

[0162] MS m/z 1006.3 [M+H].sup.+

Step 7

[0163] 1-6 (720 mg, 0.904 mmol), 2-7 (650 mg, 0.645 mmol), sodium iodide (0.406 g, 2.71 mmol) and boric acid (17 mg, 0.271 mmol) were added to a reaction flask. After the flask was purged with argon three times, NMP (2.1 mL) was added. The reaction mixture was allowed to react at room temperature until the reaction was complete. The reaction mixture was directly used in the next step without treatment.

[0164] MS m/z 1767.5 [M].sup.+

Step 8

[0165] NMP (0.7 mL) was added to the above reaction mixture, and the reaction mixture was cooled to 0? C. After phosphorus trichloride (0.103 mL, 1.17 mmol) was added, the mixture was allowed to react at 0? C. until the reaction was complete. The reaction mixture was triturated with 5% aqueous sodium hydrogen sulfite solution (20 mL) in an ice-water bath and filtered. After the filter cake was dissolved with DCM (20 mL), the solution was dried over anhydrous sodium sulfate, filtered and concentrated to dryness to give 2-9 (1.21 g).

[0166] MS m/z 1751.4 [M].sup.+

Step 9

[0167] 2-9 (1.21 g, 0.69 mmol) was dissolved in anisole (0.2 mL) and trifluoroacetic acid (0.8 mL), and the solution was allowed to react at room temperature until the reaction was complete. The reaction mixture was cooled to 0? C., then triturated with MTBE (20 mL) in an ice-water bath and filtered. The filter cake was rinsed with MTBE and dried to give a crude product (0.82 g). The crude product was purified by reversed-phase preparative HPLC to give compound 2 (107 mg).

[0168] MS m/z 993.1 [M+H].sup.+

Example 3

[0169] ##STR00034## ##STR00035##

Step 1

[0170] To a reaction flask were added 3-1 (3.7 g, 28.0 mmol) (synthesized according to the method of Angewandte Chemie International Edition, 2010, 49, 7208-7212), 1-1 (26.4 g, 61.6 mmol) and DCM (316 mL), and the compounds were dissolved by stirring. The solution was cooled in an ice-water bath. DIPEA (11.9 g, 92.4 mmol) and DMTMM (23.2 g, 84.0 mmol) were added. The mixture was warmed to room temperature and stirred until the reaction was complete. The reaction mixture was filtered, and the filtrate was washed with water, dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The crude product was purified by flash chromatography on silica gel to give the product 3-2 (21.8 g, yield: 82%).

[0171] LC/MS (ESI): m/z 953.1 [M+H].sup.+

[0172] .sup.1H-NMR (400 MHz, CDCl.sub.3) ?: 1.63-1.94 (m, 4H), 3.19-3.27 (m, 3H), 3.42-3.47 (m, 2H), 3.61-3.65 (m, 1H), 3.67-3.82 (m, 14H), 3.90-3.97 (m, 1H), 4.88-5.04 (m, 8H), 6.77-6.97 (m, 12H), 7.22-7.40 (m, 8H).

Step 2

[0173] 3-2 (21.7 g, 22.75 mmol) and DCM (217 mL) were added to a reaction flask. The flask was purged with nitrogen and cooled in an ice-water bath. TEA (8.06 g, 79.63 mmol) was added dropwise. MsCl (7.82 g, 68.25 mmol) was dissolved in 8 mL of DCM and the solution was slowly added dropwise to the reaction system. After the addition, the mixture was stirred in an ice-water bath until the reaction was complete. The reaction was quenched with water and the pH was adjusted to 8 with saturated aqueous NaHCO.sub.3 solution. The organic phase was separated and collected. The aqueous phase was extracted with DCM. The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to dryness to give the product 3-3 (24.0 g, yield: 102%).

[0174] LC/MS (ESI): m/z 1031.2 [M+H].sup.+

Step 3

[0175] To a reaction flask were added 3-3 (23.9 g, 22.75 mmol) and THF (240 mL), and the compound was dissolved by stirring. LiBr (5.93 g, 68.25 mmol) was added. After the addition, the reaction was heated to 60? C. and stirred. After the reaction was complete, the reaction mixture was concentrated to remove the solvent. Water was added, and the mixture was extracted with EA. The organic phases were combined, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated to dryness to give the product 3-4 (22.2 g, yield: 94%).

[0176] LC/MS (ESI): m/z 1015.2 [M+H].sup.+

Step 4

[0177] To a reaction flask were added 3-4 (22.1 g, 21.73 mmol) and ACN (442 mL), and the compound was dissolved by stirring. K.sub.2CO.sub.3 (10.51 g, 76.06 mmol) and pyrrolidine (4.64 g, 65.19 mmol) were then added separately. After the addition, the reaction was heated to 60? C. and stirred. After the reaction was complete, the reaction mixture was concentrated to remove the solvent. Water was added, and the mixture was extracted with a mixture of the solvents DCM and MeOH (10:1). The organic phases were combined and washed with saturated brine. The organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to dryness. The crude product was purified by normal phase column chromatography on silica gel to give the product 3-5 (16.2 g, yield: 74%).

[0178] LC/MS (ESI): m/z 1006.3 [M+H].sup.+

[0179] .sup.1H-NMR (400 MHz, DMSO-d6) ?: 1.50-1.65 (m, 4H), 1.74-1.89 (m, 4H), 2.21-2.38 (m, 3H), 2.69-2.91 (m, 3H), 2.99-3.16 (m, 4H), 3.60-3.68 (m, 2H), 3.70-3.77 (m, 12H), 4.81-5.16 (m, 8H), 6.81-7.45 (m, 20H), 8.16-8.34 (m, 1H).

Step 5

[0180] To a reaction flask were added 1-6 (5.4 g, 6.78 mmol), 3-5 (8.2 g, 8.14 mmol), NaI (3.05 g, 20.34 mmol) and boric acid (136 mg, 2.03 mmol), and the flask was purged with argon three times. NMP (27 mL) was added under an ice-water bath. After the addition, the mixture was warmed to room temperature and stirred until the reaction was complete. The reaction mixture was directly used in the next step without further purification.

[0181] LC/MS (ESI): m/z 1767.9 [M].sup.+ (ion current of the strongest response)

Step 6

[0182] The reaction mixture from the previous step was cooled to 0? C. Phosphorus trichloride (1.58 g, 11.53 mmol) was slowly added dropwise. After the addition, the mixture was stirred at 0? C. until the reaction was complete. The reaction mixture was diluted with water and filtered to give crude 3-7 (14 g). The crude product was dried in vacuo and directly used in the next step.

[0183] LC/MS (ESI): m/z 1751.5 [M].sup.+ (ion current of the strongest response)

Step 7

[0184] The crude 3-7 was dissolved in anisole (28 mL) and trifluoroacetic acid (112 mL), and the solution was allowed to react at room temperature. After the reaction was complete, the reaction mixture was cooled to 0? C., triturated with MTBE (200 mL) in an ice-water bath and filtered. The filter cake was rinsed with MTBE and dried to give a crude product (8 g). The crude product was purified by reversed-phase preparative HPLC to give the product compound 3 (3.37 g, yield: 50%).

[0185] LC/MS (ESI): m/z 993.2 [M+H].sup.+

[0186] .sup.1H-NMR (400 MHz, DMSO-d6) ?: 1.44-1.46 (m, 6H), 1.59-2.20 (m, 8H), 2.83-3.89 (m, 16H), 4.87-5.19 (m, 2H), 5.68-5.79 (m, 1H), 6.53-6.64 (m, 1H), 6.67-6.84 (m, 4H), 7.30 (s, 2H), 8.30 (s, 1H), 9.42 (br s, 3H), 10.14 (br s, 2H).

Biological Evaluation

[0187] The present disclosure is further described and explained below with reference to test examples, which are not intended to limit the scope of the present disclosure.

Test Example 1: Antifungal Activity Test

[0188] 1. The test compounds were dissolved in sterile normal saline, vortexed for uniformly mixing, and diluted in a 2-fold dilution with the sterile normal saline (11 concentration points in total). The test concentration range was 32-0.031 ?g/mL, and two duplicate wells were set for each concentration. Additional wells without drug dosing were set as growth controls.

[0189] 2. 4 ?L of the diluent was added to 196 ?L of the bacterial suspension in a 96-well plate (the bacterial count in the bacterial suspension was 2-8?10.sup.5 colony forming units/mL).

[0190] 3. After 24 h of incubation at 36? C., the MIC reading was taken with the naked eye.

[0191] The minimum inhibitory concentration (MIC) assay was performed with reference to the Clinical and Laboratory Standards Institute (CLSI) guidelines.

TABLE-US-00001 Strain (gram-negative ID-CAMHB, MIC, ?g/mL bacteria) Strain ID AMR gene Cefiderocol Compound 1 Compound 2 Compound 3 Carbapenem-resistant FDA-CDC AR- strB, OXA-65, 0.5 0.25 0.5 0.25 Acinetobacter BANK#0036 OXA-24, strA, sul2 baumannii (CRAB, FDA-CDC AR- OXA-58, OXA-100, sul2 0.125 0.125 0.125 0.125 Acinetobacter BANK#0052 baumannii) FDA-CDC AR- OXA-23, OXA-65, 4 32 4 >32 BANK#0063 OXA-24, sul2 FDA-CDC AR- OXA-58, OXA-100, 0.0625 0.125 0.25 0.125 BANK#0070 sul2 FDA-CDC AR- strB, OXA-65, 0.5 1 1 0.5 BANK#0101 OXA-24, strA, sul2 Carbapenem-resistant FDA-CDC AR- TEM-1; SHV-12; KPC-2 1 0.5 1 0.5 Enterobacteriaceae BANK#0361 (CRE, Klebsiella FDA-CDC AR- TEM-1; SHV-11; KPC-2 0.5 0.25 0.25 0.125 pneumoniae) BANK#0362 FDA-CDC AR- TEM-1; SHV-12; KPC-2 0.25 0.125 0.5 0.125 BANK#0363 FDA-CDC AR- aph(3)-Ia, dfrA12, KPC-3, 2 0.5 1 0.25 BANK#0438 OmpK35, oqxA, oqxB, SHV-11 FDA-CDC AR- KPC-3, OmpK35, oqxA, 1 1 1 0.5 BANK#0453 oqxB, SHV-11, TEM-1B Carbapenem-resistant FDA-CDC AR- aac(6)-Il, aadA1b, BCR1, 0.5 0.125 0.5 0.125 Pseudomonas BANK#0439 IMP-18, OXA-50 aeruginosa (CRPA, FDA-CDC AR- KPC-2 1 2 8 2 Pseudomonas BANK#0441 aeruginosa) FDA-CDC AR- aac(6)-Il, dfrB5, OXA-4, 1 2 4 2 BANK#0444 tet(G), VIM-2 FDA-CDC AR- aadA6, VIM-2 0.25 0.125 0.25 0.25 BANK#0457 Control strain* ATCC 25922 0.0625 0.0625 0.125 0.0625 (Escherichia coli) Control strain* ATCC 27853 0.0625 0.125 0.0625 0.25 (Pseudomonas aeruginosa)

Test Example 2: Pharmacokinetic Study of Test Compounds in Cynomolgus Monkeys

1. Sample Preparation

[0192] An appropriate amount of a test compound was weighed precisely and added to a container, and 0.9% sodium chloride injection and 0.2 M NaOH solution were added thereto in an ice bath until the test compound was completely dissolved, so that a sample solution at a concentration of 2 mg/mL was obtained followed by storage at 2-8? C. for later use.

2. Test Animals

[0193] Species and strain: cynomolgus monkey [0194] Animal grade: general grade [0195] Animal source: Guangxi Xiongsen Primate Experimental Animal Breeding Development Co., Ltd. [0196] Number of the using of laboratory animal: SYXK (Su) 2019-0012

3. Test Method

[0197] 6 cynomolgus monkeys (half male and half female) were selected and randomly divided into 3 groups with 1 animal/sex in each group. The cynomolgus monkeys were administered with the test sample at a dose of 10 mg/kg by the single intravenous infusion. Blood samples were collected from each group of animals at the following time points: before administration, and 5 min, 15 min, 0.5 h, 1 h, 2 h, 4 h, 6 h, and 8 h after administration. In the test, the concentration of each test compound in the plasma of the cynomolgus monkey was detected by the LC-MS/MS method, and the lower limits of quantification of the analysis methods for the plasma samples were all 1 ?g/mL. The plasma concentration data were analyzed by the non-compartmental analysis (NCA) of the analysis software for pharmacokinetic data, WinNonlin, pharmacokinetic parameters were calculated, and pharmacokinetic characteristics of the test compounds in cynomolgus monkeys after administration were investigated, with the results shown in the following table.

TABLE-US-00002 Animal t.sub.1/2 Tmax Cmax AUC last AUCinf Vd CL MRT Sample No. (h) (h) (ug/mL) (h*ug/mL) (h*ug/mL) (mL/kg) (mL/h/kg) (h) Compound 1 2073541 0.62 0.08 35.48 29.29 32.94 270.05 303.57 0.64 2073542 0.56 0.08 30.64 23.44 25.64 313.34 390.01 0.62 Compound 3 2073543 2.57 0.08 55.18 148.87 166.33 222.96 60.12 2.50 2073544 2.26 0.08 48.60 140.08 153.20 213.11 65.27 2.49 Cefiderocol 2073545 0.86 0.08 30.53 26.21 32.39 380.98 308.70 0.70 2073546 0.75 0.08 24.33 23.94 28.35 379.63 352.76 0.70

[0198] The compounds of the present disclosure are superior in terms of Cmax and AUC as compared to cefiderocol.

Test Example 3: Pharmacokinetic Study of Test Compound in ICR Mice

1. Sample Preparation

[0199] The formulation for administration was prepared on ice. A proper amount of the test compound was precisely weighed out, and a 90% volume of normal saline was slowly added. The mixture was ultrasonically stirred in an ice-water bath until no large pieces existed. After the mixture was well mixed, a 1 M NaOH solution was added in small portions using a pipette. After the test sample was completely dissolved as seen by the naked eye, the pH value was measured, and the solution was brought to the final volume to give the formulation for subcutaneous injection with the desired concentration.

2. Test Animals

[0200] Species and strain: ICR mice [0201] Animal grade: SPF [0202] Animal source: Medicilon: 999M-018

3. Test Method

[0203] The test sample was administered by subcutaneous injection, and the administration regimen is shown in the table below.

TABLE-US-00003 Administration Test compound Animal Test compound Concentration of Volume of Number Dose solution Administration Route of Collected Group Male Female Test compound (mg/kg) (mg/mL) (mL/kg) administration* sample 1 12 12 Cefiderocol 100 10 10 Subcutaneous Plasma injection 2 12 12 Compound 3 10 1 10 Subcutaneous Plasma injection 3 12 12 Compound 3 100 10 10 Subcutaneous Plasma injection

[0204] Administration by subcutaneous injection: 3 mice/sex/time point; blood samples were collected at 9 time points: 0.083 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 8 h and 24 h after administration. The collected blood samples were placed in EDTA-K.sub.2 anticoagulation blood collection tubes. The collected whole blood was placed on ice and centrifuged at 6800 g at 2-8? C. for 6 min to separate plasma within 1 h. After the centrifugation, the plasma was transferred to a centrifuge tube to which a plasma stabilizer had been added in advance (plasma:concentrated phosphoric acid=100:0.2; for example, 1 ?L of 10% concentrated phosphoric acid was added to 50 ?L of plasma). All the animals were subjected to careful clinical observation before administration and at each of the blood collection time points after administration, and the adverse reaction symptoms in the animals were recorded.

[0205] Test results: The pharmacokinetic data for each group are shown below.

TABLE-US-00004 Pharmacokinetic parameter Cefiderocol Compound 3 Compound 3 Dose mg .Math. kg.sup.?1 100 10 100 K.sub.el h.sup.?1 1.45 1.04 1.64 T.sub.1/2 h 0.477 0.665 0.421 t.sub.max h 0.250 0.250 0.500 C.sub.max ug .Math. mL.sup.?1 362.8 113.7 935.8 AUC.sub.0-t h .Math. ug .Math. mL.sup.?1 260.9 108.7 1102.4 AUC.sub.0-inf h .Math. ug .Math. mL.sup.?1 261.1 110.0 1104.3 AUMC.sub.0-t h .Math. h .Math. ug .Math. mL.sup.?1 150.3 80.3 871.4 AUMC.sub.0-inf h .Math. h .Math. ug .Math. mL.sup.?1 150.7 86.9 880.4 MRT.sub.PO h 0.577 0.790 0.797

[0206] The exposure of the compound of the present disclosure in mice was 4.2 times greater than that of cefiderocol.