Antimicrobial compounds and methods of use thereof

Abstract

Compounds useful in the treatment of bacterial infections, medicaments comprising the compounds, and methods of use and manufacture thereof. The compounds are believed to target protein-protein interactions involving RNA polymerase and sigma factors. The compounds show antimicrobial activity, with minimum inhibitory concentrations (MICs) as low as 1 µg / mL, demonstrating their efficacy against bacteria. In addition, these compounds demonstrate low cytotoxicity.

Description

Cross-references to related applications

[0001] This application claims priority from U.S. Provisional Patent Application No. 63 / 579,550, filed August 30, 2023, which is incorporated herein by reference in its entirety. Technical Field

[0002] This disclosure relates to compounds that can be used as antimicrobial agents, pharmaceutical compositions comprising said compounds, and methods of using them. Background Technology

[0003] Bacterial transcription is a target for antimicrobial agent discovery because rifampin and fenadine are two commercially available bacterial transcription inhibitors that target the core enzyme RNA polymerase (RNAP). However, bacterial transcription also represents an underutilized target because other transcription inhibitors have not yet been developed as antimicrobial drugs. Unfortunately, the use of rifampin and fenadine has led to antimicrobial resistance (AMR) due to the mutable structure of RNAP, which limits the potential of other natural products as bacterial transcription inhibitors in drug development.

[0004] Bacterial transcription is regulated by a number of small proteins called transcription factors. These bind to RNAPs to form various transcription complexes that regulate RNA synthesis. Therefore, targeting protein-protein interactions (PPIs) in bacterial transcription could inhibit this fundamental biological process in bacteria and provide a pathway for the discovery of novel antibiotics.

[0005] PPIs have historically been undrugatable due to the challenge of designing small molecules that target large, flat interaction surfaces of proteins. However, some key binding sites on PPI interaction surfaces are small enough to be suitable for drug design. Several PPI inhibitors have also been discovered as antimicrobial agents. Nevertheless, there remains a need to develop novel compounds with improved antimicrobial properties. Summary of the Invention

[0006] In the first aspect, this article provides a compound of Formula 1: 1 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 3; n is an integer selected from 1 to 3; p is an integer selected from 1 to 4; X is -S-, -O-, -C(R)2- or -N(R-); Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl or -(CR 6 2) q Y, where q is an integer selected from 0 to 6; R 6 Each occurrence of R is independently selected from hydrogen, alkyl, and cycloalkyl; or two occurrences of R. 6 Together with the covalently bonded carbon atoms, they form 3-6 membered cycloalkyl groups; and Y is an alkynyl, optionally carbonyl-substituted cycloalkyl, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; R 3 It is hydrogen or alkyl; R 4 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; and R 5 Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R. The condition is that the compound of formula 1 is not: .

[0008] In some implementations, X is -S- or -O-.

[0009] In some implementations, R 5 Each time it appears, it is independently selected from alkyl, haloalkyl, perhaloalkyl, perhaloalkoxy, and halides.

[0010] In some implementations, R 4 Each time it appears, it is independently selected from hydrogen, fully halogenated alkyl, nitrile, nitro, -C(O)R, -C(O)OR, -C(O)N(R)2, -S(O)2R and -S(O)2N(R)2.

[0011] In some implementations, R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2 and -C(O)N(R)2.

[0012] In some embodiments, the compound has Formula 2: 2 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 3; n is an integer selected from 1 to 2; p is an integer selected from 1 to 2; X is -S- or -O-; Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1It is alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl or -(CR 6 2) q Y, where q is an integer selected from 0 to 6; R 6 Each occurrence of R is independently selected from hydrogen, alkyl, and cycloalkyl; or two occurrences of R. 6 Together with the covalently bonded carbon atoms, they form 3-6 membered cycloalkyl groups; and Y is an alkynyl, optionally carbonyl-substituted cycloalkyl, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2 and -C(O)N(R)2; R 3 It is hydrogen or alkyl; R 4 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; R 5 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; R 7It is a fully haloalkyl, nitrile, nitro, -C(O)R, -C(O)OR, -C(O)N(R)2, -S(O)2R or -S(O)2N(R)2; and R 8 Each time it appears, it is independently selected from alkyl, haloalkyl, perhaloalkyl, perhaloalkoxy, and halides.

[0014] In some implementations, R 1 It is a cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, or -(CH2) group. q Y, where q is an integer selected from 0-6; and Y is an alkynyl group, a cycloalkyl group optionally substituted with a carbonyl moiety, a heteroaryl group, a heterocycloalkyl group, a 3,3-difluoropyrrolidone-1-yl group, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R.

[0015] In some embodiments, the compound has Formula 3: 3 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 3; X is -S- or -O-; Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is a cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, or -(CH2) group. q Y, where q is an integer selected from 0-6, and Y is an alkynyl group, a cycloalkyl group optionally substituted with a carbonyl moiety, a heteroaryl group, a heterocycloalkyl group, or a 3,3-difluoropyrrolidone-1-yl group. N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2 and -C(O)N(R)2; R 3 It is hydrogen or C1-C3 alkyl; R 4 It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; and R 5 It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R.

[0017] In some implementations, R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, -OR and -SR.

[0018] In some implementations, m is 1 or 2; and R 2 Each time it appears, it is independently selected from hydrogen, alkyl, fully haloalkyl, halide and -O-(C1-C6 alkyl).

[0019] In some implementations, R 1 It is an aralkyl group or -(CH2) q Y, where q is an integer selected from 0-5, and Y is an alkynyl group, a cycloalkyl group optionally substituted with a carbonyl moiety, a heteroaryl group, a heterocycloalkyl group, or a 3,3-difluoropyrrolidone-1-yl group. N-phthalimide, arginine, -CN, -OR, -C(O)R, -C(O)OR, -N(R)C(O)R, -N(R)C(O)OR, -S(O)2R, -S(O)2N(R)2 or -N(R)2.

[0020] In some embodiments, the compound has Formula 4: 4 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 2; Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is an aralkyl group or -(CH2) q Y, where q is an integer selected from 0-5, and Y is an alkynyl, -OR, -CN, cycloalkyl optionally substituted with a carbonyl group, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, -C(O)OR, N -phthalimide, arginine, -CN, -OR, -C(O)R, -N(R)C(O)OR, -S(O)2R, -S(O)2N(R)2 or -N(R)2; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, fully haloalkyl, halide or -O- (C1-C6 alkyl); R 3 It is hydrogen or C1-C3 alkyl; R 4 It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; and R 5It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R.

[0022] In some implementations, R 3 R 4 and R 5 Each is hydrogen.

[0023] In some implementations, R 1 Selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0024] In some implementations, R 2Each time it appears, it is independently selected from hydrogen, methyl, fluoride, bromide, chloride, -CF3 or -OCH3.

[0025] In some implementations, R 1 Selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and ; R 2 Each occurrence is independently selected from hydrogen, methyl, fluoride, bromide, chloride, -CF3, or -OCH3; and R 3 It is hydrogen.

[0026] In some embodiments, the compound has Formula 5: 5 Or a pharmaceutically acceptable salt thereof, wherein R 1 Selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0028] In some implementations, R 1 Selected from: , , , , , , , , and .

[0029] In some embodiments, the compound is selected from: , , , , And its pharmaceutically acceptable salts.

[0030] In a second aspect, this document provides a pharmaceutical composition comprising the compound described herein and at least one pharmaceutically acceptable carrier or pharmaceutically acceptable excipient.

[0031] In a third aspect, this article provides a method for treating a bacterial infection in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the compound described herein.

[0032] In some implementations, the bacterial infection is caused by Gram-positive bacteria.

[0033] In some embodiments, the bacterial infection is caused by Enterococcus faecalis, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus pyogenes, Streptococcus agalactiae, Enterococcus faecalis, Staphylococcus aureus, Streptococcus pneumoniae, methicillin-resistant Staphylococcus aureus, or vancomycin-resistant Staphylococcus aureus.

[0034] In some embodiments, the bacterial infection is caused by antibiotic-resistant Staphylococcus aureus infection or antibiotic-resistant Streptococcus pneumoniae infection. Attached Figure Description

[0035] The above and other objectives and features of this disclosure will become apparent when taken in conjunction with the accompanying drawings and the following description of this disclosure.

[0036] Figure 1 The structure of A) the E. coli RNAP holoenzyme crystal complex and B) in σ 2.2 Interaction at the interface with β'CH.

[0037] Figure 2 The structures of A) C3 and C3-005 are depicted; B) the docking model (surface view) of C3 and β'CH are shown.

[0038] Figure 3 The antimicrobial activity (MIC μg / mL) of the selected compounds against clinically important Gram-positive pathogens was described. EFAE: Enterococcus faecalis ATCC 19433, SAUR a Staphylococcus aureus ATCC 25923, SAUR b Staphylococcus aureus ATCC 29213, SEPI: Staphylococcus epidermidis ATCC 12228, SSAP: Staphylococcus saprophyticus ATCC 15305, SPNE a Streptococcus pneumoniae ATCC 49619, SPNE b Streptococcus pneumoniae strain TCH8431 (HM-145), SPNE c : Streptococcus pneumoniae strain NP112 (NR-19213), SPYO: Streptococcus pyogenes (Group A Streptococcus) ATCC 19615, SAGA: Streptococcus agalactiae (Group B Streptococcus) ATCC 12386. Cip: Ciprofloxacin, Gen: Gentamicin, Oxa: Oxacillin, Van: Vancomycin.

[0039] Figure 4The antimicrobial activity (MIC µg / mL) of the selected compounds against methicillin-resistant Staphylococcus aureus (MRSA) strains was characterized. HA-MRSA: 232ST5: W-232 ST5, 233ST5: W-233 ST5, 234ST5: W-234 ST5, 235ST5: W-235 ST5, BAA-43: Staphylococcus aureus ATCC BAA-43, BAA-44: Staphylococcus aureus ATCC BAA-44, ST239: HA-MRSA 07B082243 ST239. CA-MRSA: 45ST59: W-45 ST59, 46ST59: W-46ST59, 47ST59: W-47 ST30, 48ST217: W-48 ST217, ST22: CA-MRSA 11B086169 ST22, USA300: CA-MRSA strain USA300. Laboratory: 1199B: Staphylococcus aureus SA-1199B, ANT4: Staphylococcus aureus SA-ANT4', APH2: Staphylococcus aureus SA-APH2”-AAC6', APH3: Staphylococcus aureus SA-APH3', pUL5054: Staphylococcus aureus SA-RN4220-pUL5054. Commercially available antibiotics: Cip: Ciprofloxacin, Gen: Gentamicin, Oxa: Oxacillin, Van: Vancomycin. § M20-01: CUHK_M20_01, M20-02: CUHK_M20_02, M20-03: CUHK_M20_03, M20-04: CUHK_M20_04, M20-05: CUHK_M20_05.

[0040] Figure 5The antimicrobial activity (MIC μg / mL) of the selected compounds against VRSA strains was described. VRSA: NR-46410: Staphylococcus aureus strain HIP11714, NR-46411: Staphylococcus aureus strain HIP11983, NR-46412: Staphylococcus aureus strain HIP13170, NR-46413: Staphylococcus aureus strain HIP13419, NR-46414: Staphylococcus aureus strain HIP14300, NR-46415: Staphylococcus aureus strain HIP15178, NR-46416: Staphylococcus aureus strain HIP14300, NR-46415: Staphylococcus aureus strain HIP15178, NR-46416: Staphylococcus aureus strain HIP15178. Strains AIS2006032, NR-46417: Staphylococcus aureus strains AIS2006045, NR-46418: Staphylococcus aureus strain 71080, NR-46419: Staphylococcus aureus strain AIS080003, NR-46420: Staphylococcus aureus strain AIS1000505, NR-46421: Staphylococcus aureus strain AIS1001095, NR-46422: Staphylococcus aureus strain AID1001123. Ciprofloxacin (Cip), Gen: Gentamicin, Oxa: Oxacillin, Van: Vancomycin.

[0041] Figure 6 Epifluorescence microscopy images of A) Bacillus subtilis BS1048; B) Bacillus subtilis BS1048 treated with rifampicin; C) Bacillus subtilis BS1048 treated with chloramphenicol; and D) Bacillus subtilis BS1048 treated with 5 days (from left to right: 1, 2, 4, and 8 μg / mL).

[0042] Figure 7 Depicts the use of AutoDock Vina, docked to β Compounds of CH. (A) with β The 5d and 7d binding conformation of the CH complex. In β Details of the interaction between CH and 5d (B) and 7d (C). Images generated by PyMol. Detailed Implementation

[0043] The following terms will be used to describe the present invention. In the absence of specific definitions set forth herein, the terms used to describe the present invention should be interpreted in accordance with their common meanings as understood by one of ordinary skill in the art.

[0044] Throughout this disclosure, unless the context otherwise requires, the word "comprise" or its variations such as "comprises" or "comprising" will be understood to imply inclusion of the specified integers or groups of integers, but not to exclude any other integers or groups of integers. It should also be noted that in this disclosure, and particularly in the claims and / or paragraphs, terms such as "comprises," "comprised," "comprising," etc., may have the meanings accorded to them under U.S. patent law; for example, they may mean "includes," "included," "including," etc.; and terms such as "consisting essentially of" and "consists essentially of" have the meanings accorded to them under U.S. patent law, for example, they allow elements not expressly stated, but exclude elements found in the prior art or elements affecting the essential or novel features of the invention.

[0045] Furthermore, throughout this disclosure and the claims, unless the context otherwise requires, the word “include” or variations such as “includes” or “including” will be understood to imply inclusion of the specified integer or group of integers, but not to exclude any other integer or group of integers.

[0046] The use of the singular in this document includes the plural (and vice versa), unless otherwise expressly stated. Furthermore, where the term "about" is used before a quantity value, the present teaching also includes the specific quantity value itself, unless otherwise expressly stated. As used herein, the term "about" means a variation of ±10%, ±7%, ±5%, ±3%, ±1%, or ±0% of the nominal value, unless otherwise specified or inferred.

[0047] As used herein, the terms “treat,” “treating,” “treatment,” etc., refer to the reduction or improvement of a condition / disease and / or its associated symptoms. It will be understood that, although not excluded, treating a condition or disease does not require the complete elimination of the condition, disease, or its associated symptoms. In some implementations, treatment includes the prevention of a condition or disease and / or its associated symptoms. As used herein, the terms “prevention” or “prevent” refer to any action that inhibits or at least delays the occurrence of a condition, disease, or its associated symptoms. Prevention may include primary, secondary, and tertiary levels of prevention, wherein: a) primary prevention avoids the occurrence of disease; b) secondary prevention activities aim to treat disease early, thereby increasing opportunities for intervention to prevent disease progression and the onset of symptoms; and c) tertiary prevention reduces the negative effects of existing disease by restoring function and reducing disease-related complications.

[0048] As used herein, the term "subject" refers to an animal, typically a mammal or human, who will be or has been the subject of treatment, observation, and / or experimentation. When the term is used in conjunction with the administration of the compounds described herein, the subject is already the subject of treatment, observation, and / or administration of the compounds described herein.

[0049] As used herein, the term "therapeutic effective amount" means the amount of a compound or agent that elicits a biological and / or medical response in a cell culture, tissue system, subject, animal, or human, a response sought by an investigator, veterinarian, clinician, or physician, including the reduction of symptoms of the disease, condition, or ailment being treated.

[0050] The term "composition" is intended to include products containing specified amounts of specified ingredients, as well as any products produced directly or indirectly from combinations of specified amounts of specified ingredients.

[0051] The term "pharmaceutically acceptable carrier" refers to a medium used to prepare the desired dosage form of a compound. Pharmaceutically acceptable carriers may include one or more solvents, diluents or other liquid media; dispersants or suspending agents; surfactants; isotonic agents; thickeners or emulsifiers; preservatives; solid binders; lubricants, etc. Remington's Pharmaceutical Sciences, 15th edition, EW Martin (Mack Publishing Co., Easton, Pa., 1975) and Handbook of Pharmaceutical Excipients, 3rd edition, AH Kibbe ed. (American Pharmaceutical Association, 2000) disclose various carriers for formulating pharmaceutical compositions and known techniques for their preparation.

[0052] As used herein, unless otherwise specified, the terms “halogenated” or “halogenated” include fluorinated, chlorinated, brominated, or iodinated compounds.

[0053] As used herein, "alkyl" refers to a straight-chain or branched saturated hydrocarbon group. Examples of alkyl groups include methyl-, ethyl-, propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, sec-butyl, tert-butyl), pentyl (e.g., 1-methylbutyl, 2-methylbutyl, isopentyl, tert-pentyl, 1,2-dimethylpropyl, neopentyl, and 1-ethylpropyl), hexyl, etc. In various embodiments, the alkyl group may have 1-40 carbon atoms (i.e., C1-40 alkyl), for example, 1-30 carbon atoms (i.e., C1-30 alkyl). In some embodiments, the alkyl group may have 1-6 carbon atoms and may be referred to as "lower alkyl". Examples of lower alkyl groups include methyl, ethyl, propyl (e.g., n-propyl and isopropyl), and butyl (e.g., n-butyl, isobutyl, sec-butyl, tert-butyl). In some embodiments, the alkyl group may optionally be substituted, as described herein. Alkyl groups are not typically substituted by another alkyl, alkenyl, or ynyl group.

[0054] As used herein, "alkenyl" refers to a straight-chain or branched alkyl group having one or more carbon-carbon double bonds. Examples of alkenyl groups include vinyl, propenyl, butenyl, pentenyl, hexenyl, butadienyl, pentadienyl, hexadienyl, etc. The one or more carbon-carbon double bonds can be internal (e.g., in 2-butene) or terminal (e.g., in 1-butene). In various embodiments, the alkenyl group can have 2-40 carbon atoms (i.e., C2-40 alkenyl), for example 2-20 carbon atoms (i.e., C2-20 alkenyl). In some embodiments, the alkenyl group can be substituted, as described herein. The alkenyl group is generally not substituted by another alkenyl, alkyl, or ynyl group.

[0055] As used herein, "cycloalkyl" itself, or as part of another substituent, unless otherwise specified, means a monocyclic hydrocarbon having 3-12 carbon atoms in a ring system and includes hydrogen, straight-chain, branched, and / or cyclic substituents. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, etc.

[0056] As used herein, "heteroatoms" refers to atoms of any element other than carbon or hydrogen, and includes, for example, nitrogen, oxygen, silicon, sulfur, phosphorus, and selenium.

[0057] As used herein, the term "heterocyclic alkyl" includes references to saturated heterocyclic moieties having 3, 4, 5, 6, or 7 ring carbon atoms and 1, 2, 3, 4, or 5 ring heteroatoms selected from nitrogen, oxygen, phosphorus, and sulfur. The group can be a polycyclic ring system, but is more commonly monocyclic. The term includes references to groups such as azirrobutyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, oxadiazolidinyl, pyrazolyl, imidazolyl, indolizidyl, piperazinyl, thiazolyl, morpholinyl, thiomorpholinyl, quinolizidyl, etc.

[0058] As used herein, "aryl" refers to an aromatic monocyclic or polycyclic ring system in which two or more aromatic rings are fused together (i.e., have a common bond) or at least one aromatic monocyclic ring is fused to one or more cycloalkyl and / or heterocyclic alkyl rings. An aryl group may have 6-24 carbon atoms in its ring system (e.g., C6-24 aryl), and may include multiple fused rings. In some embodiments, a polycyclic aryl group may have 8-24 carbon atoms. Any suitable ring position of the aryl group may be covalently linked to a defined chemical structure. Examples of aryl groups having only aromatic carbon rings include phenyl, 1-naphthyl (bicyclic), 2-naphthyl (bicyclic), anthracene (tricyclic), phenanthrene (tricyclic), and pentaphenyl (pentacyclic) groups. Examples of polycyclic systems in which at least one aromatic carbide ring is fused to one or more cycloalkyl and / or heteroalkyl rings include, but are not limited to, benzo[i](x) derivatives of cyclopentane (i.e., indenyl, which is a 5,6-bicyclocycloalkyl / aromatic ring system), benzo[i](x) derivatives of cyclohexane (i.e., tetrahydronaphthalene, which is a 6,6-bicyclocycloalkyl / aromatic ring system), benzo[i](x) derivatives of imidazoline (i.e., benzimidazolinyl, which is a 5,6-bicyclocycloalkyl / aromatic ring system), and benzo[i](x) derivatives of pyran (i.e., benzopyranyl, which is a 6,6-bicyclocycloalkyl / aromatic ring system). Other examples of aryl groups include benzodioxane-hexyl, benzodioxane-pentyl, benzodihydropyranyl, indololinyl, etc. In some embodiments, the aryl group may optionally be substituted.

[0059] The term "aralkyl" refers to an alkyl group that has been substituted with an aryl group.

[0060] As used herein, "heteroaryl" refers to an aromatic monocyclic or polycyclic ring system containing at least one cyclic heteroatom selected from oxygen (O), nitrogen (N), sulfur (S), silicon (Si), and selenium (Se), wherein at least one ring in the ring system is aromatic and contains at least one cyclic heteroatom. Polycyclic heteroaryls include those having two or more heteroaryl rings fused together, and those having at least one monocyclic heteroaryl ring fused to one or more aromatic carbocyclic, non-aromatic, and / or non-aromatic cyclic heteroalkyl rings. Heteroaryls as a whole may have, for example, 5-24 ring atoms and contain 1-5 cyclic heteroatoms (i.e., 5-20 membered heteroaryls). Heteroaryls may be attached to a defined chemical structure at any heteroatom or carbon atom that results in a stable structure. Typically, heteroaryl rings do not contain OO, SS, or SO bonds. However, one or more N or S atoms in a heteroaryl group may be oxidized (e.g., pyridine N-oxide, thiophene S-oxide, thiophene S,S-dioxide). Examples of heteroaryl groups include, for example, 5- or 6-membered monocyclic and 5- to 6-membered bicyclic ring systems shown below, where T is O, S, NH, N-alkyl, N-aryl, N-(arylalkyl) (e.g., N-benzyl), SiH2, SiH(alkyl), Si(alkyl)2, SiH(arylalkyl), Si(arylalkyl)2, or Si(alkyl)(arylalkyl). Examples of such heteroaryl rings include pyrroloyl, furanyl, thiophene, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazolyl, tetrazolyl, pyrazolyl, imidazolyl, isothiazolyl, thiazolyl, thiadiazolyl, isoxazolyl, oxazolyl, oxadiazolyl, indolyl, isoydinolyl, benzofuranyl, benzothiophene, quinolinyl, 2-methylquinolinyl, isoquinolinyl, quinoxalinyl, quinazolinyl, benzotriazolyl, benzimidazolyl, benzothiazolyl, Benzisothiazolyl, benzisoxazolyl, benzoxadiazolyl, benzoxazolyl, cenolinyl, 1H-inzolyl, 2H-inzolyl, indoleazinyl, isobenzofuranyl, naphridinyl, phthalazinyl, pteridinyl, purineyl, oxazolopyridinyl, thiazopyridinyl, imidazopyridinyl, furanopyridinyl, thienopyridinyl, pyridinopyrimidinyl, pyridinopyrazinyl, pyridinopyridazinyl, thienothiazoyl, thienooxazolyl, thienoimidazoyl, etc. Further examples of heteroaryl groups include 4,5,6,7-tetrahydroindolyl, tetrahydroquinolinyl, benzothienopyridinyl, benzofuranopyridinyl, etc. In some embodiments, the heteroaryl group may be substituted, as described herein. In some embodiments, the heteroaryl group may optionally be substituted.

[0061] The term "optional substitution" refers to a chemical group, such as alkyl, cycloalkyl, aryl, etc., in which one or more hydrogen atoms may be replaced by substituents as described herein, such as halogens, azides, alkyl, aralkyl, alkenyl, alkynyl, cycloalkyl, hydroxyl, alkoxy, amino, nitro, mercapto, imino, amide, phosphonate, hypophosphite, carbonyl, carboxyl, silyl, ether, alkylthio, sulfonyl, sulfonamide, ketone, aldehyde, ester, heterocyclic, aromatic or heteroaromatic moiety, -CF3, -CN, etc.

[0062] The term "nitro" is generally accepted in the art and refers to -NO2; the term "halogen" is generally accepted in the art and refers to -F, -Cl, -Br, or -I; the term "mercapto" is generally accepted in the art and refers to -SH; the term "hydroxyl" means -OH; and the terms "sulfonyl" and "sulfone" are generally accepted in the art and refer to -SO2-. "Halide" refers to the corresponding anion of the halogen.

[0063] The symbol "in chemical structure" "or" "or" "or" "" indicates the position where a specified chemical structure is bonded to another chemical structure.

[0064] As used herein, the term "pharmaceutically acceptable salt" means that, within reasonable medical judgment, it is suitable for use in contact with the tissues of a subject without excessive toxicity, irritation, allergic reactions, etc., and is commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences (1977) 66:1-19. Pharmaceutically acceptable salts of the compounds provided herein include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, or with organic acids such as acetic acid, oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipic acid salts, alginate salts, ascorbate salts, aspartate salts, benzenesulfonate salts, besylate salts, benzoate salts, hydrogen sulfate salts, borate salts, butyrate salts, camphorate salts, camphor sulfonate salts, citrate salts, cyclopentanepropionate salts, diglucuronate salts, dodecyl sulfate salts, ethanesulfonate salts, formate salts, fumarate salts, glucohepanoate salts, glycerophosphate salts, glucuronate salts, hemisulfate salts, heptanate salts, and hexanoate salts. The organic acids that can be derived from salts include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, dihydroxynaphthalate, pectate, persulfate, 3-phenylpropionate, phosphate, picrate, neopentanoate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, etc. In some embodiments, the organic acids that can be derived from salts include, for example, acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.

[0065] Pharmaceutically acceptable salts derived from suitable alkalis include alkali metals, alkaline earth metals, ammonium, and nitrogen. + (C 1-4Alkyl)4 salts. Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc. Further pharmaceutically acceptable salts, where appropriate, include non-toxic ammonium, quaternary ammonium, and amine cations formed using anti-charge ions such as halide, hydroxide, carboxyl, sulfate, phosphate, nitrate, lower alkyl sulfonates, and aryl sulfonates. Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines (including naturally occurring substituted amines), cyclic amines, basic ion exchange resins, etc., such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, and ethanolamine. In some embodiments, pharmaceutically acceptable base addition salts are selected from ammonium, potassium, sodium, calcium, and magnesium salts.

[0066] This disclosure provides a compound of Formula 1: 1 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 3; n is an integer selected from 1 to 3; p is an integer selected from 1 to 4; A is a part with the following structure: ; X is -S-, -O-, -C(R)2- or -N(R-); Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl or -(CR 6 2) q Y, where q is an integer selected from 0 to 6; R 6 Each occurrence of R is independently selected from hydrogen, alkyl, and cycloalkyl; or two occurrences of R. 6 Together with the covalently bonded carbon atoms, they form 3-6 membered cycloalkyl groups; and Y is an alkynyl, optionally carbonyl-substituted cycloalkyl, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; R 3 It is hydrogen or alkyl; R 4 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; and R 5 Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R. The condition is that the compound of formula 1 is not: .

[0068] The compounds described herein may include isomers at different positions indicated by bonds that are not connected to the vertices of the chemical structure, for example, using the model structure illustration below: .

[0069] In this model structure, where the valence allows, the group R can be attached to any atom on the ring structure, i.e., carbon 2, 3, 4, 5 or 6 in the above structure.

[0070] In some implementations, m is an integer selected from 1-3 or 1-2. In some implementations, m is 1, 2, or 3.

[0071] In some implementations, n is an integer selected from 1-3 or 1-2. In some implementations, n is 1, 2, or 3.

[0072] In some implementations, p is an integer selected from 1-4, 1-3, 2-3, or 1-2. In some implementations, p is 1, 2, or 3.

[0073] When m is 1 or 2, A can be represented by a subset of the following: , , , , , , and , Where R 2 and R 3 Each as defined in any of the implementations described herein.

[0074] In some implementations, A is selected from the following: , , , , , , , , , , , , , , , , , , and , Where R 2 and R 3 Each as defined in any of the implementations described herein.

[0075] X can be -S-, -O-, -C(R)2-, or -N(R)-, wherein each R is independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form a 3-6 membered cycloalkyl group. In some embodiments, X is -S-, -O-, -CH2-, -NH-, or -N(C1-C6 alkyl)-. In some embodiments, X is -S-, -O-, -CH2-, -N(Me)-, or -NH-. In some embodiments, X is -S- or -O-.

[0076] In some embodiments, R is selected independently each time it appears, from hydrogen, alkylcycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups.

[0077] R 1 It can be C2-C 12 Alkyl, C3-C 12 Alkyl, C4-C 12 Alkyl, C3-C 10 Alkyl, C3-C8 alkyl, C3-C6 alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, or -(CR2) q Y, where R 6 Each time it appears, it is independently selected from hydrogen, C1-C6 alkyl, and C1-C6 alkyl, q is an integer selected from 0-6, and Y is alkynyl, optionally carbonyl-substituted cycloalkyl, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R. In some embodiments, R 1 It is cycloalkyl, aralkyl, or -(CR2). q Y, where q is an integer selected from 1 to 4, and Y is a cycloalkyl, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, or -N(R)2. An exemplary R 1 Parts include, but are not limited to: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0078] R 2 Each occurrence can be independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, and -C(O)N(R)2. In some embodiments, R 2 Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, -OR, and -SR. In some embodiments, at least one R 2 It is a C1-C6 alkyl, C1-C3 alkyl, C1-C2 alkyl, C1-C3 perhaloalkyl, C1-C2 perhaloalkyl, halide, -O-(C1-C6 alkyl), -O-(C1-C3 alkyl), or -O-(C1-C2 alkyl). In some embodiments, at least one R 2 It can be methyl, -CF3, fluoride, chloride, bromide, or -OMe.

[0079] R 3 It can be hydrogen, C1-C6 alkyl, C1-C5 alkyl, C1-C4 alkyl, C1-C3 alkyl, or C1-C2 alkyl. In some embodiments, R 3 It is hydrogen, methyl, or ethyl. In R 3 When R is an alkyl group, the ester can be metabolized or hydrolyzed under physiological conditions to form the corresponding carboxylic acid, i.e., where R is an alkyl group. 3It is hydrogen.

[0080] R 4 Each occurrence can be independently selected from hydrogen, perhaloalkyl, nitrile, nitro, -C(O)R, -C(O)OR, -C(O)N(R)2, -S(O)2R, and -S(O)2N(R)2. In some embodiments, at least one R 4 It is a nitrile, nitro, -C(O)OR, -C(O)N(R)2, -S(O)2R, or -S(O)2N(R)2. In some embodiments, at least one R 4 It is a nitro group.

[0081] R 5 Each occurrence can be independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, and -C(O)N(R)2. In some embodiments, at least one or at least two R... 5 It is a C1-C3 alkyl, C1-C3 haloalkyl, C1-C3 perhaloalkyl, halide, nitrile, or nitro group. In some embodiments, at least two R groups are present. 5 It is methyl, halomethyl, perhalomethyl, fluoride, chloride, or nitrile. In some embodiments, at least two R... 5 It is methyl, FCH2-, F2CH-, CF3-, chloride or nitrile.

[0082] In some embodiments, the compound has Formula 5: 5 Or a pharmaceutically acceptable salt thereof, wherein R 1 Selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

[0084] In some embodiments, the compound is selected from: , , , , , , , , , , , , , , , , And its pharmaceutically acceptable salts.

[0085] This disclosure also provides a pharmaceutical composition comprising the compound described herein and at least one pharmaceutically acceptable excipient and / or pharmaceutically acceptable carrier.

[0086] The compounds described herein and their pharmaceutically acceptable salts may be administered to subjects, alone or in combination with a pharmaceutically acceptable carrier or diluent, in a pharmaceutical composition, in accordance with standard pharmaceutical practice. The compounds may be administered parenterally. Parenterally administration includes intravenous, intramuscular, intraperitoneal, subcutaneous, and topical administration.

[0087] Therefore, this disclosure provides pharmaceutically acceptable compositions comprising a therapeutically effective amount of the compound described herein, formulated with one or more pharmaceutically acceptable carriers (additives) and / or diluents. The pharmaceutical compositions of this disclosure may be specifically formulated for administration in liquid form, including those suitable for: (1) parenteral administration, such as as a sterile solution or suspension, for example, via intravenous administration.

[0088] As illustrated herein, certain embodiments of the compounds described herein may contain a basic functional group, such as an amino group, and thus be able to form pharmaceutically acceptable salts with pharmaceutically acceptable acids. In this regard, the term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic and organic acid addition salt of the compounds of this disclosure. These salts may be prepared in situ during the manufacture of an application medium or dosage form, or by reacting a purified compound of the invention in its free basic form with a suitable organic or inorganic acid, and separating the resulting salt during subsequent purification. Representative salts include bromides, chlorides, sulfates, bisulfates, carbonates, bicarbonates, nitrates, acetates, valerates, oleates, palmitates, stearates, laurates, benzoates, lactates, phosphates, toluenesulfonates, citrates, maleates, fumarates, succinates, tartrates, naphthalates, methanesulfonates, glucono-p-ethylates, lacturonates, and laurylsulfonates, etc.

[0089] Pharmaceutically acceptable salts of the compounds disclosed herein include conventional non-toxic salts or quaternary ammonium salts of the compounds, such as those derived from non-toxic organic or inorganic acids. For example, such conventional non-toxic salts include those derived from inorganic acids, such as hydrochloric acid, hydrobromic acid, sulfuric acid, aminosulfonic acid, phosphoric acid, nitric acid, etc.; and salts prepared from organic acids, such as acetic acid, propionic acid, succinic acid, glycolic acid, stearic acid, lactic acid, malic acid, tartaric acid, citric acid, ascorbic acid, palmitic acid, maleic acid, hydroxymaleic acid, phenylacetic acid, glutamic acid, benzoic acid, salicylic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, fumaric acid, toluenesulfonic acid, methanesulfonic acid, ethanedisulfonic acid, oxalic acid, hydroxyethylsulfonic acid, etc.

[0090] In other cases, the compounds described herein may contain one or more acidic functional groups, thus enabling them to form pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these cases, the term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic or organic base addition salt of the compounds of the present invention. These salts can be prepared in situ during the manufacture of the administration medium or dosage form, or by reacting a purified compound in its free acid form with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable primary, secondary, or tertiary organic amine. Representative alkali metal or alkaline earth metal salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts. Representative organic amines that can be used to form base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc.

[0091] Wetting agents, emulsifiers and lubricants (such as sodium lauryl sulfate and magnesium stearate), as well as colorants, releasing agents, coating agents, sweeteners, flavoring agents and aromas, preservatives, solubilizers, buffers and antioxidants may also be present in the composition.

[0092] Methods for preparing a pharmaceutical product comprising the compound described herein include the steps of combining the compound described herein with a carrier and optionally one or more excipients. Generally, a formulation is prepared by uniformly and tightly combining the compound described herein with a liquid carrier (liquid formulation), the liquid carrier followed by lyophilization (powder formulation, for reconstitution with sterile water, etc.), or a finely powdered solid carrier, or both, and then shaping or packaging the product if desired.

[0093] The pharmaceutical compositions suitable for parenteral administration according to this disclosure comprise one or more of the compounds described herein, combined with one or more pharmaceutically acceptable sterile isotonic aqueous or non-aqueous solutions, dispersions, suspensions or emulsions, or sterile powders (which may be reconstituted into sterile injectable solutions or dispersions immediately prior to use), which may contain sugars (e.g., sucrose), alcohols, nonionic surfactants (e.g., Tween 20), antioxidants, buffers, antibacterial agents, chelating agents, solutes or suspending agents or thickeners that make the formulation isotonic with the blood of the intended recipient.

[0094] Examples of suitable aqueous and non-aqueous carriers that can be used in the pharmaceutical compositions of this disclosure include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, etc.) and suitable mixtures thereof, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. For example, in the case of dispersions, appropriate flowability can be maintained by using a coating material such as lecithin, by maintaining the desired particle size, and by using a surfactant.

[0095] These compositions may also contain adjuvants such as preservatives, wetting agents, emulsifiers, and dispersants. Prevention of microbial action on the compounds of this disclosure can be ensured by including various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, etc. Isotonic agents, such as sugars, sodium chloride, etc., may also be included in the composition. Furthermore, prolonged absorption of injectable drug forms can be achieved by including agents that delay absorption, such as aluminum monostearate and gelatin.

[0096] On the other hand, this article provides a method for treating a bacterial infection in a subject in need, the method comprising the step of administering a therapeutically effective amount of the compound described herein to the subject.

[0097] Exemplary bacterial infections include, but are not limited to, pneumonia, otitis media, sinusitis, bronchitis, tonsillitis, and mastoiditis associated with infections of *Streptococcus pneumoniae*, *Haemophilus influenzae*, *Moraxella catarrhalis*, *Staphylococcus aureus*, *Enterococcus faecalis*, *Enterococcus faecium*, *Enterococcus pyogenes*, *Staphylococcus epidermidis*, *Staphylococcus hemolyticus*, or *Peptostreptococcus* species; pharyngitis, rheumatic fever, and glomerulonephritis associated with infections of *Streptococcus pyogenes*, group C and group G streptococci, *Corynebacterium diphtheriae*, or *Actinomyces hemolyticus*; respiratory infections associated with infections of *Mycoplasma pneumoniae*, *Legionella pneumophila*, *Streptococcus pneumoniae*, *Haemophilus influenzae*, or *Chlamydia pneumoniae*; and infections caused by *Staphylococcus aureus*, *Staphylococcus hemolyticus*, *Enterococcus faecalis*, *Streptococcus pneumoniae*, *Enterococcus faecalis*, *Streptococcus pneumoniae*, *Haemophilus influenzae*, or *Chlamydia pneumoniae*. Enterococcus faecalis and Enterococcus tenuisae, including strains resistant to known antimicrobial agents (such as, but not limited to, β-lactams, vancomycin, aminoglycosides, quinolones, chloramphenicol, tetracyclines, and macrolides), causing blood and tissue infections, including endocarditis and osteomyelitis; uncomplicated skin and soft tissue infections and abscesses associated with Staphylococcus aureus, coagulase-negative staphylococci (i.e., Staphylococcus epidermidis, Staphylococcus hemolyticus, etc.), Streptococcus pyogenes, Streptococcus agalactiae, Group CF Streptococci (small colony streptococci), viridans streptococci, Corynebacterium microphyllum, Clostridium species, or Bartonella henselae, as well as puerperal fever; infections associated with Staphylococcus aureus and coagulase-negative staphylococci. Uncomplicated acute urinary tract infections associated with infection with *Enterococcus* species or *Enterococcus* spp.; urethritis and cervicitis; sexually transmitted infections associated with *Chlamydia trachomatis*, *Haemophilus ducreyi*, *Treponema pallidum*, *Ureaplasma urealyticum*, or *Neisseria gonorrhoeae*; toxin-related illnesses associated with *Staphylococcus aureus* (food poisoning and toxic shock syndrome) or *Streptococcus* spp. A, B, and C; ulcers associated with *Helicobacter pylori* infection; systemic febrile syndrome associated with *Relapsing fever* infection; Lyme disease associated with *Borrelia burgdorferi* infection; infections associated with *Chlamydia trachomatis*, *Neisseria gonorrhoeae*, *Staphylococcus aureus*, *Streptococcus pneumoniae*, *Streptococcus pyogenes*, *Haemophilus influenzae*, or *Listeria* species. Infection-related conjunctivitis, keratitis, and dacryocystitis; disseminated mycobacterial complex (MAC) disease associated with infection with Mycobacterium avium or intracellular mycobacteria; infections caused by Mycobacterium tuberculosis, Mycobacterium leprae, Mycobacterium paratuberculosis, Mycobacterium kansas, or Mycobacterium cypriniforme; gastroenteritis associated with infection with Campylobacter jejuni; intestinal protozoan infections associated with infection with Cryptosporidium species; odontogenic infections associated with infection with Streptococcus viridans; persistent cough associated with infection with Bordetella pertussis; gas gangrene associated with infection with Clostridium perfringens or Bacteroides species; and atherosclerosis or cardiovascular disease associated with infection with Helicobacter pylori or Chlamydia pneumoniae.Bacterial infections and related conditions that can be treated or prevented in animals include the following: bovine respiratory diseases associated with infections of Pasteurella multocida, Pasteurella multocida, Mycoplasma bovis, or Bordetella species; bovine intestinal diseases associated with infections of Escherichia coli or protozoa (i.e., coccidia, Cryptosporidium, etc.); dairy cow mastitis associated with infections of Staphylococcus aureus, Streptococcus lactis, Streptococcus agalactiae, Streptococcus dysgalactiae, Klebsiella species, Corynebacterium species, or Enterococcus species; swine respiratory diseases associated with infections of Actinobacillus pleuropneumoniae, Pasteurella multocida, or Mycoplasma species; swine intestinal diseases associated with infections of Escherichia coli, Lawsonia intracellularis, Salmonella species, or Spirochetes swine dysenteriae; and diseases associated with infections of *Fusobacterium*. Bovine foot rot associated with infection of fungal species; bovine metritis associated with infection of *Escherichia coli*; bovine hairy warts associated with infection of *Fusobacterium necrophorum* or *Bacteroides nodosum*; bovine conjunctivitis associated with infection of *Moraxella bovis*; bovine premature birth and abortion associated with infection of protozoa (i.e., *Neospora*); urinary tract infections associated with infection of *Escherichia coli* in dogs and cats; skin and soft tissue infections associated with infection of *Staphylococcus epidermidis*, *Staphylococcus intermedius*, coagulase-negative staphylococci*, or *Pasteurella multocida* in dogs and cats; and dental or oral infections associated with infection of species of *Alcaligenes*, *Bacteroides*, *Clostridium*, *Enterobacter*, *Eubacterium*, *Peptostreptococcus*, *Porphyromonas*, or *Prevotella* in dogs and cats. Other bacterial infections and conditions associated with said infections that can be treated or prevented according to the method of the present invention are mentioned in JP Sanford et al., “The Sanford Guide To Antimicrobial Therapy,” 26th edition, (Antimicrobial Therapy, Inc., 1996).

[0098] In some embodiments, the bacteria are selected from Streptococcus agalactiae, Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus saprophyticus, and Enterococcus faecalis. In some embodiments, the bacteria are SAUR ATCC 25923, SAUR ATCC 29213, or SPNE ATCC 49619.

[0099] The subject can be a canine, feline, bovine, equine, non-human primate, or human. In some embodiments, the subject is a human.

[0100] In some embodiments, the compounds described herein are bactericidal or bacteriostatic.

[0101] On the other hand, this article provides the use of the compounds described herein in the preparation of medicaments for treating bacterial infections.

[0102] On the other hand, this document provides a method for treating a bacterial infection in a subject in need, the method comprising the step of co-administering a therapeutically effective amount of the compound described herein and an antibacterial agent to the subject. The antibacterial agent may be any antibacterial agent known in the art.

[0103] Based on structural information and biochemical confirmation, the major binding sites between RNAP and the housekeeping sigma (σ) factor were investigated. The latter is a small protein highly conserved in bacteria (i.e., σ in Gram-negative bacteria). A and σ in Gram-negative bacteria 70 And it binds to RNAP to initiate bacterial transcription by recognizing a DNA promoter. Figure 1 A). The amino acids important for binding σ region 2.2 in the RNAP β' subunit clamp helical region (β'CH) were identified. Figure 1 B) Subsequently, computer-based screening of structure-based drug design and drug-like compound libraries was used to identify several hit compounds, including C3. Other RNAP-σ PPI inhibitors were also identified.

[0104] Based on compound C3 ( Figure 2 A) Structural modification and antimicrobial activity testing were conducted. The benzoic acid moiety was found to be a key structure for binding β'CH and antimicrobial activity. Therefore, this class of novel antimicrobial compounds was named sigmacidin, aiming to embody structure-based design by mimicking the σ factor and the key structure of benzoic acid. Since the substituent on the left-hand benzene ring significantly enhances antimicrobial activity (C3-005), the carbonyl linker of C3-005 was examined for modification to a sulfonamide group in this study. The antimicrobial activity and mechanism, cytotoxicity, and pharmacokinetic properties of the sulfonamide analog were investigated.

[0105] In previous studies, carbonyl, methylene, and aminomethylene linkers provided better antimicrobial activity than analogs with amide linkers. In this paper, the sulfonamide moiety was chosen as a linker to connect the diphenyl sulfide and benzoic acid moieties to examine the structural requirements of the linker. Additionally, based on the pharmacophore docking model, the benzoic acid moiety is clamped by β'CH R278 and R281 (…). Figure 1 The substituted benzoic acid (B and 2B) was designed to probe the small half-pocket formed by β'CH and to examine the effect of the substitution on the benzoic acid on antimicrobial activity.

[0106] Compared to amides, sulfonamides possess more acidic protons suitable for substitution reactions. This study used different group modifications of the sulfonamide nitrogen to determine their effects on the antimicrobial activity of novel analogs.

[0107] Compounds 5a–5j were synthesized as shown in Scheme 1. 4-Chloro-3-nitrobenzenesulfonyl chloride was condensed with the corresponding amine to provide compounds 3a–3j in yields of 50%–75%. The -Cl group was then substituted with 3,4-dichlorobenzenethiophenol to provide intermediates 4a–4j in yields of 70%–95%. Finally, the methyl ester group was hydrolyzed to provide compounds 5a–5j. Compound 5b was synthesized in the first stage for antimicrobial activity testing to investigate the effect of substitution at the sulfonamide nitrogen.

[0108]

[0109] Scheme 1. Synthetic routes for compounds 5a–5j. Reagents and conditions: a) aniline, pyridine, dichloromethane (DCM), 0 a) overnight, 50% – 75%; b) 3,4-dichlorobenzylthiophenol, NaOAc, EtOH, reflux, 6 h, 70% – 90%; c) NaOH, H2O / dioxane, 50 C, overnight.

[0110] Preliminary antimicrobial activity tests revealed that compound 5b, methylated at the nitrogenous site of the sulfonamide, exhibited reduced antibacterial activity. This suggests that modification of the sulfonamide group may affect the activity. As shown in Scheme 2, sulfonamide intermediates 4c–4f were reacted with a series of organic bromides in DMF in the presence of K₂CO₃ to yield 6a–6h in yields of 60%–90%. Hydrolysis of the methyl esters yielded compounds 7a–7k.

[0111]

[0112] Scheme 2. Synthetic route for compounds 7a–7k. Reagents and conditions: a) R 1 Br, K2CO3, DMF, 80 a) overnight, 60% – 90%; b) NaOH, H2O / dioxane, 50 C, overnight.

[0113] As described in Scheme 3, alkylamine derivatives were also synthesized. Alkylation of 4c with 1,4-dibromobutane in the presence of K₂CO₃ yielded compound 8 in 72% yield, which was further amination with various secondary amines to provide compounds 9a–9d. The methyl ester was finally hydrolyzed to provide… N -alkylamines 10a – 10d.

[0114]

[0115] Scheme 3. Synthetic route for compounds 10a–10d. Reagents and conditions: a) 1,4-dibromobutane, K₂CO₃, DMF, 50 C) Overnight, 72%; b) Amine, K2CO3, DMF, 40 c) NaOH, H2O / dioxane, 50% – 90% C, overnight.

[0116] The antimicrobial activity of all synthesized compounds against *Streptococcus pneumoniae* and *Staphylococcus aureus* (two of the WHO priority pathogens for antibiotic development) was tested. Compounds 5a–5j were first tested under standard CLSI conditions, and antimicrobial activity was expressed as MIC values. 5a confirmed antimicrobial activity against both bacteria, but *Streptococcus pneumoniae* was more sensitive to treatment with 5a than *Staphylococcus aureus* (MIC 2 μg / mL vs. 8–16 μg / mL), similar to what was shown with previously designed and synthesized sigmacidin compounds with carbonyl, methylene, and aminomethylene linkers. As mentioned above, the antimicrobial activity of 5b decreased significantly to MICs of 64–128 μg / mL compared to 5a due to the methyl substitution on the sulfonamide nitrogen of 5a. Despite the sulfonamide nitrogen remaining unsubstituted, 5c–5j showed preservation or enhancement against some or all of the tested bacteria, regardless of the presence of electron-withdrawing or electron-donating substituents. These results differ from previous findings that electron-donating groups on the right-hand benzene ring decrease antimicrobial activity while electron-withdrawing groups increase it. Maximum activity was observed when 5-trifluoro-2-benzoic acid was bonded to the sulfonamide linker (5d), leading to enhanced antimicrobial activity with MICs of 2 and 4 μg / mL against Streptococcus pneumoniae and Staphylococcus aureus, respectively. Note that the clogP values ​​of these compounds ranged from 6.73 to 8.14, and antimicrobial activity did not show a correlation with clogP values. This phenomenon, unlike previous results with different linkers, suggests that membrane permeability may not be particularly significantly correlated with the antimicrobial activity of sulfonamide derivatives of sigmacidin.

[0117] Table 1. Antimicrobial activity and clogP values ​​of synthesized compounds 5a-5j

[0118] SPNE: Streptococcus pneumoniae ATCC 49619, SAUR a Staphylococcus aureus ATCC 29213, SAUR b Staphylococcus aureus ATCC 29523 Although 5b showed reduced antimicrobial activity, the substituent at the sulfonamide nitrogen site was observed to affect antimicrobial activity, and the effect of substitution was intended to be fully investigated. The antimicrobial activities of 7a–7k and 10a–10d against *Streptococcus pneumoniae* and *Staphylococcus aureus* were tested. Surprisingly, all these compounds confirmed enhanced antimicrobial activity against some or all of the tested bacterial strains. Both aryl and aliphatic substituents were acceptable, confirming the potential for structural modification at this site. Furthermore, some compounds, such as 7a, 7b, and 10a–10d, showed superior antimicrobial activity against *Staphylococcus aureus* compared to *Streptococcus pneumoniae*. This is the first time this has been observed compared to other sigmacidin derivatives. Maximum activity against *Staphylococcus aureus* was obtained with 7b, with a MIC of 2 μg / mL. In these compounds, when the same cyclopropylmethyl substituent is present on the sulfonamide nitrogen, we also observed that the electron-donating group (5-Me in 7a) on the right-hand benzene ring contributes slightly more to the antimicrobial activity than the electron-withdrawing group (4-F in 7f, 4-Cl in 7i, and 5-F in 7j).

[0119] Table 2. Antimicrobial activity and clogP values ​​of the synthesized compounds 7a-7k and 10a-10d

[0120] SPNE: Streptococcus pneumoniae ATCC 49619, SAUR a Staphylococcus aureus ATCC 29213, SAUR b Staphylococcus aureus ATCC 29523 After obtaining preliminary antimicrobial activity, the selected compounds were screened against a wider range of bacterial species from the WHO priority pathogen list used to guide the development of novel antibiotics, such as Enterococcus faecalis, Staphylococcus aureus, and Streptococcus pneumoniae. Additionally, clinically complex pathogens, such as Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus pyogenes, and Streptococcus agalactiae, were tested to confirm the clinical potential of the compounds described herein.

[0121] Figure 3The results described confirm the broad antimicrobial potential of the compounds described herein against a wide range of Gram-positive bacteria, including clinically complex pathogens such as *Staphylococcus epidermidis*, *Staphylococcus saprophyticus*, *Streptococcus pyogenes*, and *Streptococcus agalactiae*, with most having MICs ranging from 2 to 8 μg / mL. Notably, compounds 5d, 7a, 7b, and 7d exhibit robust antimicrobial activity against a variety of Gram-positive bacteria, with impressive MICs as low as 1 μg / mL against *Streptococcus pneumoniae*. Notably, 5d showed significant efficacy against *Streptococcus* spp. (MIC 1–2 μg / mL), while 7a demonstrated significant efficacy against *Staphylococcus epidermidis* (MIC 2 μg / mL), showcasing the potential of our compounds in the treatment of bacterial infections.

[0122] Methicillin-resistant Staphylococcus aureus (MRSA) strains represent some of the most challenging bacterial strains to treat due to antibiotic resistance, including resistance to vancomycin, currently available as a “last-line” antimicrobial agent. To evaluate the efficacy of our compounds against MRSA strains, their activity was tested against a range of representative clinical isolates of hospital-acquired (HA-) and community-acquired (CA-) MRSA strains, including vancomycin-resistant Staphylococcus aureus (VRSA) strains.

[0123] HA-MRSA strains include W-231 ST45, W-232 ST5, W-233 ST5, W-234 ST-5, W-235 ST5, ATCC BAA-43, ATCC BAA-44, and ST239. CA-MRSA strains, on the other hand, include W-45 ST59, W-46 ST59, W-47 ST30, W-48 ST217, ST22, ST338, and USA300. Several MRSA strains known as M20 are also included (N. Barua, et al., Comparative Study of Two-Dimensional (2D) vs. Three-Dimensional (3D) Organotypic Kertatinocyte-Fibroblast Skin Models for Staphylococcus aureus (MRSA) Infection, Int J Mol Sci, 23 (2021)). To further test the compounds described herein, laboratory strains with known resistance mechanisms were included, such as SA-APH2”-AAC6’, SA-APH3’, and SA-ANT4’, which are resistant to aminoglycosides due to the presence of aminoglycoside-modifying enzymes (AME). Also included were SA-RN4220-pUL5054, which exhibits resistance to macrolides, and SA-1199B, which is known to be resistant to fluoroquinolones.

[0124] like Figure 4 As shown, the compounds demonstrated antimicrobial activity against the tested MRSA strains, with the most potent MIC as low as 2 μg / mL. This efficacy surpasses currently used antibiotics such as ciprofloxacin, gentamicin, and oxacillin, especially for treating specific strains. Notably, most of the selected MRSA strains are resistant to oxacillin (the current first-line antibiotic for MRSA infections in the United States) and other common drugs such as the protein synthesis inhibitor gentamicin. Therefore, the antimicrobial activity of these compounds against MRSA strains with various individual resistance mechanisms indicates a different mode of action that does not overlap with existing antibiotics. Among the compounds tested, 5d, 7b, 7c, 7d, and 7i maintained consistent MIC values ​​ranging from 2 to 8 μg / mL against a wide range of MRSA strains, particularly the aminoglycoside-resistant strain SA-APH3', highlighting their potential for further development as a treatment for MRSA infections.

[0125] The VRSA strains used were provided by the Staphylococcus aureus Antimicrobial Resistance Network (NARSA), allocated by BEIResources, NIAID, NIH, and managed by ATCC (Manassas, Virginia, United States). These included: NR-46410 (Staphylococcus aureus strain HIP11714), NR-46411 (Staphylococcus aureus strain HIP11983), NR-46412 (Staphylococcus aureus strain HIP13170), NR-46413 (Staphylococcus aureus strain HIP13419), NR-46414 (Staphylococcus aureus strain HIP14300), NR-46415 (Staphylococcus aureus strain HIP15178), NR-46416 (Staphylococcus aureus strain AIS 2006032), and NR-46417 (Staphylococcus aureus strain AIS 2006032). 2006045), NR-46418 (Staphylococcus aureus strain 71080), NR-46419 (Staphylococcus aureus strain AIS 080003), NR-46420 (Staphylococcus aureus strain AIS 1000505), NR-46421 (Staphylococcus aureus strain AIS 1001095) and NR-46422 (Staphylococcus aureus strain AID 1001123).

[0126] like Figure 5 As shown, several compounds exhibited significant antimicrobial activity against the tested VRSA strains, with MICs as low as 2 μg / mL, in contrast to the fact that most strains were resistant to or desensitized to cell wall synthesis inhibitors vancomycin and oxacillin. Five compounds—5d, 7b, 7c, 7d, and 7i—consistently demonstrated antibacterial activity against VRSA strains, with MICs ranging from 2 to 8 μg / mL. Among these, 5d, 7c, and 7d demonstrated particularly superior antibacterial activity, with 5d being highly effective in inhibiting strains NR-46418 and NR-46421 (MIC 2 μg / mL). Therefore, these compounds demonstrate efficacy against a range of VRSA strains with multiple resistance mechanisms, strongly supporting their clinical potential and guaranteeing further development as potential novel therapeutic options for the increasingly refractory and severe infections caused by VRSA.

[0127] In summary, the C3 compounds in this series exhibit significant antibacterial activity against a range of pathogenic bacteria, including notoriously difficult-to-treat strains such as MRSA and VRSA. Compared to other Gram-positive bacteria, the unsubstituted sulfonamide compounds (5a-5j) maintained strong antimicrobial activity against Streptococcus pneumoniae, while substitution at the sulfonamide nitrogen site enhanced the overall antimicrobial activity of the resulting compounds (7a-7k) against Staphylococcus aureus, as confirmed by tests against MRSA and VRSA strains. Of the 12 compounds tested, 5d and 7d stood out, exhibiting the highest antibacterial activity against a variety of bacterial strains resistant to currently available antibiotics.

[0128] The inhibitory activity of representative sulfonamide compounds as PPI inhibitors was examined. A luminescence-based protein complementation assay was previously developed. By fusing RNAP β'CH and σ into two complementary fragments of luciferase, respectively, the luminescent signal released from the reformation of luciferase was reduced due to inhibition of the β'CH-σ PPI when a β'CH-σ PPI inhibitor was used in this assay. The results are shown in Table 3. All compounds tested showed inhibitory activity against the β'CH-σ PPI.

[0129] Table 3. IC50 of the selected compounds inhibiting β'CH-σ PPI 50

[0130] Note the IC being tested. 50 The value was not correlated with antimicrobial activity. This phenomenon has been observed in previous studies, as the latter is related to multiple factors, such as cell membrane permeability and efflux effects, which significantly affect antimicrobial activity. Furthermore, the water solubility of the compound can also affect the experimental process, thus influencing the IC50 value. 50 Therefore, this assay should be used as a qualitative method to confirm the molecular mechanism of a compound.

[0131] The subcellular effects of compound 5d on RNAP function were examined using epifluorescence microscopy. Bacillus subtilis strain BS1048 exhibited green fluorescence, which is due to the addition of green fluorescent protein (GFP) tagging to RNAP. Figure 6 As shown in A, GFP-labeled RNAP is localized to the bacterial nucleoid. Chloramphenicol, which targets the ribosome, does not affect RNAP function, while rifampin, which targets RNAP, can diffuse the transcription focus. Figure 6 B), and chloramphenicol, which targets the ribosome, has no effect on it. Figure 6 C). When compound 5d was added, similar to rifampicin-treated cells, a fluorescence signal was observed to diffuse from the nucleoid in a concentration-dependent manner. Figure 6 D) indicates that 5d, as an RNAP-σ PPI inhibitor, also affects bacterial transcription, as shown in previous studies.

[0132] like Figure 7 As shown, for 5d and 7d with β The calculated binding conformation of the CH complex was used for molecular docking. As previously mentioned, the benzoic acid moieties of both ligands are located in a positively charged region formed by residues including Arg275 and Arg278, while the 3,4-dichlorophenyl ring matches into a pouch formed by hydrophobic residues. Additionally, the 5d carboxyl group and β... Two hydrogen bonds are formed between Arg278 (O…H 2.2Å) and Asn274 (O…H 3.4 Å) of CH. Additionally, an H bond is formed between the nitro group and Arg275 (O…H 2.8 Å). Figure 7 B). Figure 7 C shows in 7d and β Interactions between CH groups. The 7d carboxyl group interacts with Arg275 (O…H 2.1 Å) and Arg278 (O…H 2.2 Å) via hydrogen bonds, respectively. Simultaneously, Arg278 also interacts with the nitro group (O…H 2.9 Å). Overall, both compounds interact with β-hydroxyl groups. The key interactions of CH residues (including Arg275 and Arg278) show similar binding energies.

[0133] 2.7. Cytotoxicity The selected compounds were tested for cytotoxicity against human HepG2 hepatocellular carcinoma and A549 lung cancer cell lines. All tested compounds demonstrated near-non-cytotoxicity against both human cancer cell lines (Table 4), indicating their potential for further therapeutic development.

[0134] Table 4. Cytotoxicity and Therapeutic Index of the Selected Compounds

[0135] a Through CC 50 / ½ MIC calculation b DDP: Cisplatin Indole derivatives have been previously developed as RNAP-σ PPI inhibitors. In this paper, we investigated the structural diversity of sulfonamide derivatives of C3-005 (sigmacidin) and confirmed their antimicrobial activity, mechanism of action, cytotoxicity, and pharmacokinetic properties. The results indicate that sulfonamide derivatives of C3-005 have the potential for further development. Specifically, the sulfonamide nitrogen provides additional and suitable substitution positions without diminishing antimicrobial activity; on the contrary, substituted derivatives demonstrated enhanced antimicrobial activity against Staphylococcus aureus, including MRSA and VRSA. Further structurally diverse sigmacidin derivatives with excellent antimicrobial activity are expected to enrich the molecular pool of RNAP-σ PPI inhibitors for biomedical research and drug development.

[0136] 4.1.1. General Method A glass slide (silicone F) that can be visualized under UV light 254 All reactions were monitored by thin-layer chromatography (TLC). Rapid chromatography was performed using silica gel (200–300 mesh). Commercially available reagents and anhydrous solvents were used without the need for further purification. All reported yields are isolated yields. 1 H NMR (400 MHz) and 13 C NMR (100 MHz) spectra were measured on a BRUKERAVANCE-III spectrometer, with TMS as an internal standard. Chemical shifts are expressed as δ (ppm), and coupling constants ( J (Measurements are in Hz.) High-resolution MS spectra were measured by electrospray ionization using a QTOF-2 micromass spectrometer. HPLC analysis was performed on an Agilent 1260 HPLC instrument.

[0137] 4.1.2. General procedure for synthesizing intermediate 3a-j (Scheme 1) General procedure for synthesizing compounds 3a–3o Pyridine (7.5 mmol) was added to a stirred solution of aniline (1 mmol) in dichloromethane (DCM, 5 mL) at 0 °C. Then, 4-chloro-3-nitrobenzenesulfonyl chloride (1.2 mmol) in DCM (5 mL) was slowly added via syringe. The solution was warmed to room temperature and stirred for an additional 24 hours. The mixture was diluted with DCM (10 mL) and washed successively with 1 M HCl and brine. The organic phase was dried over Na₂SO₄ and purified by chromatography to provide the title compound.

[0138] 2-((4-chloro-3-nitrophenyl)sulfonamido)methyl benzoate (3a) Yellow solid (208 mg, 56%). 1 H NMR (400 MHz, CDCl3) δ 10.81 (s, 1H), 8.31(d, J = 2.0 Hz, 1H), 7.95 (ddd, J = 10.9, 8.2, 1.6 Hz, 2H), 7.71 (d, J = 8.3Hz, 1H), 7.62 (d, J = 8.5 Hz, 1H), 7.57 – 7.51 (m, 1H), 7.14 (t, J = 7.6 Hz, 1H), 3.90 (s, 3H).

[0139] 2-((4-chloro-N-methyl-3-nitrophenyl)sulfonamido)methyl benzoate (3b) White solid (227 mg, 59%). 1 H NMR (400 MHz, CDCl3) δ 8.12 (d, J = 2.0 Hz, 1H), 7.91 (dd, J = 7.7, 1.6 Hz, 1H), 7.77 (dd, J = 8.4, 2.0 Hz, 1H), 7.66 (d, J = 8.4 Hz, 1H), 7.52 (td, J = 7.7, 1.6 Hz, 1H), 7.46 (td, J = 7.4, 0.9 Hz, 1H), 7.10 (dd, J = 7.8, 0.7 Hz, 1H), 3.81 (s, 3H), 3.34 (s, 3H).

[0140] 2-((4-chloro-3-nitrophenyl)sulfonamido)-5-methylbenzoate (3c) Red solid (196 mg, 51%). 1 H NMR (400 MHz, CDCl3) δ 10.55 (s, 1H), 8.27(d, J = 1.9 Hz, 1H), 7.90 (dd, J = 8.4, 2.0 Hz, 1H), 7.75 (s, 1H), 7.61 (dd,J = 8.3, 6.4 Hz, 2H), 7.34 (dd, J = 8.4, 1.2 Hz, 1H), 3.88 (s, 3H).

[0141] 4-Chloro-2-((4-chloro-3-nitrophenyl)sulfonamido)methyl benzoate (3d) White solid (223 mg, 55%). 1 H NMR (400 MHz, CDCl3) δ 10.94 (s, 1H), 8.36(d, J = 2.0 Hz, 1H), 8.00 (dd, J = 8.4, 2.0 Hz, 1H), 7.93 (d, J = 8.6 Hz, 1H), 7.77 F(d, J = 1.8 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.13 (dd, J = 8.5,1.8 Hz, 1H), 3.94 (s, 3H).

[0142] 2-((4-chloro-3-nitrophenyl)sulfonamido)-4-fluorobenzoate (3e) White solid (202 mg, 52%). 1 H NMR (400 MHz, CDCl3) δ 11.08 (s, 1H), 8.34(d, J = 1.9 Hz, 1H), 8.04 – 7.95 (m, 2H), 7.67 (d, J = 8.5 Hz, 1H), 7.46 (dd, J = 10.4, 2.3 Hz, 1H), 6.87 – 6.76 (m, 1H), 3.92 (s, 3H).

[0143] 5-Fluoro-2-((4-fluoro-3-nitrophenyl)sulfonamido)methyl benzoate (3f) Brown solid (208 mg, 56%). 1 H NMR (400 MHz, CDCl3) δ 10.44 (s, 1H), 8.26(d, J = 2.1 Hz, 1H), 7.88 (dd, J= 8.5, 2.1 Hz, 1H), 7.74 (dd, J = 9.1, 4.7Hz, 1H), 7.67 – 7.58 (m, 2H), 7.30 – 7.24 (m, 1H).

[0144] 2-((4-chloro-3-nitrophenyl)sulfonamido)-4-(trifluoromethyl)benzoate (3g) Yellow solid (272 mg, 62%). 1 H NMR (400 MHz, CDCl3) δ 10.93 (s, 1H), 8.36(d, J = 2.0 Hz, 1H), 8.13 (d, J = 8.2 Hz, 1H), 8.03 (s, 1H), 7.99 (dd, J =8.5, 2.1 Hz, 1H), 7.69 (d, J = 8.4 Hz, 1H), 7.40 (d, J = 8.2 Hz, 1H), 3.98 (s, 3H).

[0145] 2-((4-chloro-3-nitrophenyl)sulfonamido)-4-methoxybenzoate (3h) White solid (200 mg, 50%). 1 H NMR (400 MHz, CDCl3) δ 11.04 (s, 1H), 8.34(d, J = 2.1 Hz, 1H), 7.96 (dd, J = 8.5, 2.1 Hz, 1H), 7.89 (d, J = 8.9 Hz, 1H), 7.64 (d, J = 8.5 Hz, 1H), 7.22 (d, J = 2.4 Hz, 1H), 6.62 (dd, J = 8.9,2.4 Hz, 1H), 3.87 (s, 3H), 3.86 (s, 3H).

[0146] 2-((4-chloro-3-nitrophenyl)sulfonamido)-4,5-dimethoxybenzoate (3i) Red solid (250 mg, 58%). 1H NMR (400 MHz, CDCl3) δ 10.56 (s, 1H), 8.26(d, J = 2.1 Hz, 1H), 7.85 (dd, J = 8.5, 2.1 Hz, 1H), 7.60 (d, J = 8.5 Hz, 1H), 7.31 (d, J = 11.0 Hz, 2H), 3.97 (s, 3H), 3.85 (s, 6H).

[0147] 4-Bromo-2-((4-chloro-3-nitrophenyl)sulfonamido)methyl benzoate (3j) White solid (297 mg, 66%). 1 H NMR (400 MHz, CDCl3) δ 10.88 (s, 1H), 8.34(d, J = 2.0 Hz, 1H), 7.97 (dd, J = 8.5, 2.1 Hz, 1H), 7.91 (d, J = 1.5 Hz, 1H), 7.82 (d, J = 8.6 Hz, 1H), 7.67 (d, J = 8.4 Hz, 1H), 7.28 (d, J = 1.8 Hz, 1H), 3.91 (s, 3H).

[0148] 1-((4-chloro-3-nitrophenyl)sulfonamido)cyclopropane-1-carboxylic acid methyl ester (3k) Yellow solid (241 mg, 72%). 1 H NMR (400 MHz, CDCl3) δ 8.33 (d, J = 2.0 Hz, 1H), 8.00 (dd, J = 8.4, 2.0 Hz, 1H), 7.72 (d, J = 8.4 Hz, 1H), 5.62 (s, 1H), 3.45 (s, 3H), 1.54 (t, J = 2.7 Hz, 2H), 1.51 (d, J = 3.3 Hz, 2H).

[0149] ((4-chloro-3-nitrophenyl)sulfonyl)-L-valine methyl ester (3l) White solid (263 mg, 75%). 1 H NMR (400 MHz, CDCl3) δ 8.31 (d, J = 2.1 Hz, 1H), 7.98 (dd, J = 8.5, 2.1 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 5.23 (d, J =9.8 Hz, 1H), 3.86 (dd, J = 10.0, 4.9 Hz, 1H), 2.14 (qd, J = 12.1, 6.8 Hz, 1H), 1.02 (d, J = 6.8 Hz, 3H), 0.91 (d, J = 6.9 Hz, 3H).

[0150] ((4-chloro-3-nitrophenyl)sulfonyl)-L-phenylalanine methyl ester (3m) White solid (275 mg, 69%). 1 H NMR (400 MHz, CDCl3) δ 8.12 (d, J = 2.0 Hz, 1H), 7.77 (dd, J = 8.4, 2.1 Hz, 1H), 7.58 (d, J = 8.5 Hz, 1H), 7.27 – 7.17(m, 3H), 7.06 (dd, J = 6.2, 2.9 Hz, 2H), 5.25 (d, J = 9.4 Hz, 1H), 4.29 (ddd, J = 9.3, 7.8, 5.1 Hz, 1H), 3.70 (s, 3H), 3.15 (dd, J = 13.9, 5.0 Hz, 1H), 2.97 (dd, J = 13.9, 7.7 Hz, 1H).

[0151] ((4-chloro-3-nitrophenyl)sulfonyl)-L-alanine methyl ester (3n) White solid (229 mg, 71%). 1H NMR (400 MHz, CDCl3) δ 8.32 (d, J = 2.1 Hz, 1H), 7.98 (dd, J = 8.4, 2.1 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 5.39 (d, J =8.2 Hz, 1H), 4.16 – 4.03 (m, 1H), 3.64 (s, 3H), 1.45 (d, J = 7.2 Hz, 3H).

[0152] 4-Chloro-N-(2-(hydroxymethyl)phenyl)-3-nitrobenzenesulfonamide (3o) Pale yellow solid (257 mg, 75%). 1 H NMR (400 MHz, CDCl3) δ 8.27 (d, J = 2.0Hz, 1H), 8.24 (s, 1H), 7.89 (dd, J = 8.4, 2.1 Hz, 1H), 7.63 (d, J = 8.4 Hz, 1H), 7.51 (d, J = 8.1 Hz, 1H), 7.31 (d, J = 8.1 Hz, 1H), 7.14 (q, J = 7.2 Hz, 2H), 4.49 (d, J = 4.6 Hz, 2H), 2.09 – 2.01 (m, 1H).

[0153] General procedure for synthesizing compounds 4a–4o Compound 3a–3o (0.3 mmol), 3,4-dichlorothiophenol (0.5 mmol), NaOAc (1.5 mmol), and ethanol (EtOH, 3 ml) were added to a flask. The mixture was heated under reflux for 6 hours and then cooled to room temperature. The precipitate was collected by filtration and washed successively with small amounts of EtOH and water. The filter cake was dried under vacuum to give compound 4a–4o.

[0154] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)methyl benzoate (4a) Yellow solid (116 mg, 75%). 1H NMR (400 MHz, CDCl3) δ 10.82 (s, 1H), 8.69(d, J = 2.0 Hz, 1H), 7.96 (dd, J = 7.9, 1.2 Hz, 1H), 7.79 (dd, J = 8.7, 1.9Hz, 1H), 7.69 (d, J = 8.3 Hz, 1H), 7.65 (d, J = 1.9 Hz, 1H), 7.59 (d, J = 8.3Hz, 1H), 7.54 – 7.46 (m, 1H), 7.38 (dd, J = 8.2, 2.0 Hz, 1H), 7.10 (t, J =7.6 Hz, 1H), 6.90 (d, J = 8.7 Hz, 1H), 3.90 (s, 3H).

[0155] 2-((4-((3,4-dichlorophenyl)thio)-N-methyl-3-nitrophenyl)sulfonamido)methyl benzoate (4b) Yellow solid (100 mg, 63%). 1 H NMR (400 MHz, CDCl3) δ 8.51 (d, J = 1.9 Hz, 1H), 7.89 (dd, J = 7.6, 1.6 Hz, 1H), 7.71 (d, J = 1.9 Hz, 1H), 7.63 – 7.58(m, 2H), 7.49 (td, J = 7.7, 1.7 Hz, 1H), 7.46 – 7.40 (m, 2H), 7.08 (d, J =7.6 Hz, 1H), 6.94 (d, J = 8.6 Hz, 1H), 3.82 (s, 3H), 3.30 (s, 3H).

[0156] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (4c) Yellow solid (125 mg, 79%). 1H NMR (400 MHz, CDCl3) δ 10.57 (s, 1H), 8.65(d, J = 1.9 Hz, 1H), 7.75 (dd, J = 8.7, 1.9 Hz, 2H), 7.65 (d, J = 1.9 Hz, 1H), 7.60 (dd, J = 8.3, 5.5 Hz, 2H), 7.38 (dd, J = 8.2, 2.0 Hz, 1H), 7.31(dd, J = 8.4, 1.5 Hz, 1H), 6.88 (d, J = 8.6 Hz, 1H), 3.87 (s, 3H), 2.29 (s, 3H).

[0157] Methyl 4-chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)benzoate (4d) Yellow solid (141 mg, 86%). 1 H NMR (400 MHz, CDCl3) δ 10.92 (s, 1H), 8.72(d, J = 2.0 Hz, 1H), 7.90 (dd, J = 8.5, 3.6 Hz, 1H), 7.81 (dd, J = 8.6, 2.0Hz, 1H), 7.70 (dd, J = 21.8, 1.9 Hz, 2H), 7.60 (d, J = 8.2 Hz, 1H), 7.39 (dd, J = 8.3, 2.0 Hz, 1H), 7.07 (dd, J = 8.6, 1.8 Hz, 1H), 6.94 (d, J = 8.6 Hz, 1H), 3.91 (s, 3H).

[0158] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-fluorobenzoate (4e) Yellow solid (123 mg, 77%). 1H NMR (400 MHz, CDCl3) δ 11.08 (s, 1H), 8.73(d, J = 1.8 Hz, 1H), 8.00 (dd, J = 8.9, 6.3 Hz, 1H), 7.82 (dd, J = 8.7, 1.8Hz, 1H), 7.67 (d, J = 1.7 Hz, 1H), 7.60 (d, J = 8.3 Hz, 1H), 7.41 (ddd, J =10.1, 9.5, 2.1 Hz, 2H), 6.93 (d, J = 8.7 Hz, 1H), 6.78 (td, J = 9.0, 2.3 Hz, 1H), 3.91 (s, 3H).

[0159] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-fluorobenzoate (4f) Yellow solid (128 mg, 80%). 1 H NMR (400 MHz, CDCl3) δ 10.49 (s, 1H), 8.65(d, J = 1.8 Hz, 1H), 7.77 – 7.70 (m, 2H), 7.63 (ddd, J = 14.4, 8.3, 5.1 Hz,3H), 7.39 (dd, J = 8.3, 1.9 Hz, 1H), 7.25 – 7.21 (m, 1H), 6.90 (d, J = 8.7Hz, 1H), 3.90 (s, 3H).

[0160] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-(trifluoromethyl)benzoate (4g) Yellow solid (141 mg, 81%). 1 H NMR (400 MHz, CDCl3) δ 10.96 (s, 1H), 8.71(d, J = 1.9 Hz, 1H), 8.10 (d, J= 8.3 Hz, 1H), 7.98 (s, 1H), 7.81 (dd, J =8.7, 1.9 Hz, 1H), 7.65 (d, J = 1.8 Hz, 1H), 7.60 (d, J = 8.2 Hz, 1H), 7.43 –7.36 (m, 2H), 7.33 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.7 Hz, 1H), 3.95 (s, 1H).

[0161] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-methoxybenzoate (4h) Yellow solid (139 mg, 85%). 1 H NMR (400 MHz, CDCl3) δ 11.05 (s, 1H), 8.73(d, J = 1.9 Hz, 1H), 7.88 (d, J = 8.9 Hz, 1H), 7.79 (dd, J = 8.7, 2.0 Hz, 1H), 7.66 (d, J = 2.0 Hz, 1H), 7.60 (d, J = 8.3 Hz, 1H), 7.39 (dd, J = 8.2, 2.0 Hz, 1H), 7.23 (d, J = 2.4 Hz, 1H), 6.91 (d, J = 8.7 Hz, 1H), 6.59 (dd, J = 8.9, 2.4 Hz, 1H), 3.86 (s, 3H), 3.85 (s, 3H).

[0162] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4,5-dimethoxybenzoate (4i) Yellow solid (151 mg, 88%). 1 H NMR (400 MHz, CDCl3) δ 10.62 (s, 1H), 8.65(d, J = 1.9 Hz, 1H), 7.71 (dd,J = 8.6, 1.9 Hz, 1H), 7.66 (d, J = 1.9 Hz, 1H), 7.60 (d, J = 8.3 Hz, 1H), 7.38 (dd, J = 8.2, 1.9 Hz, 1H), 7.32 (d, J =1.4 Hz, 2H), 6.87 (d, J = 8.7 Hz, 1H), 3.96 (s, 3H), 3.85 (s, 3H), 3.84 (s, 3H).

[0163] 4-Bromo-2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)methyl benzoate (4j) White solid (170 mg, 96%). 1 H NMR (400 MHz, CDCl3) δ 10.88 (s, 1H), 8.72(d, J = 2.0 Hz, 1H), 7.89 (d, J = 1.7 Hz, 1H), 7.84 – 7.77 (m, 2H), 7.67 (d, J = 1.9 Hz, 1H), 7.59 (s, 1H), 7.40 (dd, J = 8.3, 2.0 Hz, 1H), 7.23 (dd, J =8.6, 1.7 Hz, 1H), 6.94 (d, J = 8.6 Hz, 1H), 3.91 (s, 3H).

[0164] 1-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)methyl cyclopropane-1-carboxylate (4k) Yellow solid (113 mg, 79%). 1 H NMR (400 MHz, CDCl3) δ 8.71 (d, J = 1.9 Hz, 1H), 7.82 (dd, J = 8.6, 1.9 Hz, 1H), 7.70 (d, J = 1.9 Hz, 1H), 7.62 (d, J=8.3 Hz, 1H), 7.42 (dd, J = 8.3, 2.0 Hz, 1H), 6.98 (d, J = 8.6 Hz, 1H), 3.45(s, 3H), 1.49 (q, J = 3.1 Hz, 4H).

[0165] ((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonyl)-L-valine methyl ester (4l) Yellow solid (113 mg, 79%). 1 H NMR (400 MHz, CDCl3) δ 8.66 (d, J = 1.9 Hz, 1H), 7.77 (dd, J = 8.7, 2.0 Hz, 1H), 7.69 (d, J = 1.9 Hz, 1H), 7.62 (d, J =8.3 Hz, 1H), 7.42 (dd, J = 8.3, 2.0 Hz, 1H), 6.97 (d, J = 8.7 Hz, 1H), 5.19(d, J = 8.7 Hz, 1H), 3.83 (dd, J = 8.6, 4.7 Hz, 1H), 3.55 (s, 3H), 2.09 (qt, J = 12.8, 6.5 Hz, 1H), 0.98 (d, J = 6.8 Hz, 3H), 0.88 (d, J = 6.9 Hz, 3H).

[0166] ((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonyl)-L-phenylalanine methyl ester (4m) Yellow solid (135 mg, 83%). 1 H NMR (400 MHz, CDCl3) δ 8.47 (d, J = 1.9 Hz, 1H), 7.69 (d, J = 2.0 Hz, 1H), 7.64 – 7.57 (m, 2H), 7.42 (dd, J= 8.3, 2.0Hz, 1H), 7.19 (dd, J = 4.9, 1.5 Hz, 3H), 7.03 (dd, J = 6.5, 2.7 Hz, 2H), 6.84(d, J = 8.7 Hz, 1H), 5.19 (s, 1H), 4.23 (s, 1H), 3.66 (s, 3H), 3.10 (dd, J =13.9, 5.1 Hz, 1H), 2.94 (dd, J = 13.9, 7.5 Hz, 1H).

[0167] ((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonyl)-L-alanine methyl ester (4n) Yellow solid (107 mg, 77%). 1 H NMR (400 MHz, CDCl3) δ 8.71 (d, J = 2.0 Hz, 1H), 7.81 (dd, J = 8.6, 2.0 Hz, 1H), 7.72 (d, J = 2.0 Hz, 1H), 7.64 (d, J =8.2 Hz, 1H), 7.45 (dd, J = 8.3, 2.0 Hz, 1H), 7.00 (d, J = 8.6 Hz, 1H), 5.32(d, J = 2.7 Hz, 1H), 4.09 (dd, J = 13.9, 6.9 Hz, 1H), 3.66 (s, 3H), 1.46 (d, J = 7.1 Hz, 3H).

[0168] 4-((3,4-dichlorophenyl)thio)-N-(2-(hydroxymethyl)phenyl)-3-nitrobenzenesulfonamide (4o) Yellow solid (123 mg, 85%). mp 156 – 158 ℃. 1 1H NMR (400 MHz, dimethyl sulfoxide (DMSO)- d 6) δ 9.84 (s, 1H), 8.44 (d, J= 1.7 Hz, 1H), 8.01 (d, J = 1.6 Hz, 1H), 7.85 (d, J = 8.3 Hz, 1H), 7.77 (dd, J = 8.6, 1.8 Hz, 1H), 7.65 (dd, J =8.3, 1.8 Hz, 1H), 7.44 (d, J = 7.4 Hz, 1H), 7.23 (t, J = 7.3 Hz, 1H), 7.19 –7.11 (m, 2H), 6.89 (d, J = 7.8 Hz, 1H), 5.18 (s, 1H), 4.44 (s, 2H). 13 C NMR (100 MHz, DMSO-) d 6) δ 144.6, 142.9, 139.1, 138.2, 137.3, 136.0, 134.4, 133.3, 133.1, 132.9, 131.9, 130.7, 130.2, 128.1, 127.7, 127.2, 126.1, 124.6, 59.6. HRMS (ESI): For C 19 H 13 Cl2N2O5S2, [MH] - Calculated value: 482.9648, measured value: 482.9647. HPLC purity: 98.68%.

[0169] General procedure for synthesizing compounds 5a–5n The methyl ester (0.1 mmol) of compound (4a – 4n) was used at 50 °C in 1 M NaOH (1 1) Hydrolyze overnight. The mixture was then diluted with a small amount of water and washed twice with DCM. The aqueous solution was acidified by adding 2 M HCl. The precipitate was collected by filtration and washed with water to provide the title compounds 5a–5n.

[0170] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)benzoic acid (5a) Yellow solid (26 mg, 53%). mp 243 – 245 ℃. 1 H NMR (400 MHz, DMSO- d6) δ11.57 (s, 1H), 8.55 (s, 1H), 7.99 (s, 1H), 7.90 (d, J = 8.2 Hz, 2H), 7.84 (d, J = 8.3 Hz, 1H), 7.63 (d, J = 8.1 Hz, 1H), 7.54 (t, J = 7.5 Hz, 1H), 7.46 (d, J = 8.1 Hz, 1H), 7.16 (t, J = 7.4 Hz, 1H), 7.10 (d, J = 8.6 Hz, 1H). 13 C NMR (100 MHz, DMSO-) d 6) δ 169.8, 144.6, 143.5, 139.6, 137.4, 137.1, 136.1, 134.8, 134.6, 133.3, 133.0, 132.0, 131.9, 130.4, 130.2, 124.9, 124.3, 119.9, 118.8. HRMS (ESI): For C 19 H 11 Cl2N2O6S2, [MH] - Calculated value: 496.9441, measured value: 496.9455. HPLC purity: 98.05%.

[0171] 2-((4-((3,4-dichlorophenyl)thio)-N-methyl-3-nitrophenyl)sulfonamido)benzoic acid (5b) Yellow solid (23 mg, 45%). mp 198 – 200 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ12.81 (s, 1H), 8.27 (s, 1H), 8.03 (s, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.78 (d, J = 7.0 Hz, 1H), 7.69 (t, J = 8.1 Hz, 2H), 7.55 – 7.44 (m, 2H), 7.16 (d, J =8.6 Hz, 1H), 7.08 (d, J= 7.4 Hz, 1H), 3.21 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6) δ 167.5, 144.7, 142.9, 139.3, 137.3, 136.2, 136.0, 134.5, 133.4, 133.3, 133.0, 132.8, 132.4, 131.1, 130.7, 130.1, 129.6, 129.1, 125.0, 39.4. HRMS(ESI): For C 20 H 13 Cl2N2O6S2, [MH] - Calculated value: 510.9598, measured value: 510.9595. HPLC purity: 97.12%.

[0172] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (5c) Yellow solid (31 mg, 60%). mp 128 – 130 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 11.35 (s, 1H), 8.51 (d, J = 1.9 Hz, 1H), 7.98 (d, J = 1.6 Hz, 1H), 7.84 (dd, J = 8.5, 2.7 Hz, 2H), 7.69 (s, 1H), 7.62 (dd, J = 8.3, 1.8 Hz, 1H), 7.36 (s,2H), 7.08 (d, J = 8.7 Hz, 1H), 2.24 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6) δ169.7, 144.5, 143.4, 137.4, 137.1, 136.8, 136.1, 135.3, 134.6, 133.8, 133.3, 133.0, 132.0, 131.9, 130.4, 130.2, 124.8, 120.5, 119.2, 20.5. HRMS (ESI): For C 20 H 13 Cl2N2O6S2, [MH] -Calculated value: 510.9598, measured value: 510.9607. HPLC purity: 99.67%.

[0173] 4-Chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)benzoic acid (5d) Yellow solid (27 mg, 51%). mp 110 – 112 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 12.71 (s, 1H), 8.55 (d, J = 1.9 Hz, 1H), 7.99 (d, J = 1.9 Hz, 1H), 7.93 –7.86 (m, 2H), 7.83 (d, J = 8.3 Hz, 1H), 7.63 (dd, J = 8.3, 1.9 Hz, 1H), 7.44(d, J = 1.8 Hz, 1H), 7.16 (dd, J = 8.5, 1.6 Hz, 1H), 7.12 (d, J = 8.6 Hz, 1H). 13 C NMR (100 MHz, DMSO- d 6) δ 168.8, 144.6, 143.2, 142.4, 138.7, 137.8, 137.4, 136.1, 134.5, 133.6, 133.3, 132.9, 131.8, 130.4, 130.3, 124.6, 123.4, 119.1, 118.1. HRMS (ESI): For C 19 H 10 Cl3N2O6S2, [MH] - Calculated value: 530.9051, measured value: 530.9033. HPCL purity: 95.31%.

[0174] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-fluorobenzoic acid (5e) Yellow solid (34 mg, 66%). mp 218 – 220 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 11.88 (s, 1H), 8.59 (d, J= 2.0 Hz, 1H), 8.00 (d, J = 2.0 Hz, 1H), 7.99 –7.93 (m, 2H), 7.84 (d, J = 8.3 Hz, 1H), 7.64 (dd, J = 8.3, 2.0 Hz, 1H), 7.23(dd, J = 10.8, 2.4 Hz, 1H), 7.11 (d, J = 8.7 Hz, 1H), 7.00 (td, J = 8.6, 2.4Hz, 1H). 13 C NMR (100 MHz, DMSO- d 6) δ 169.1, δ 165.3 (d, J = 251.9 Hz), 144.6,143.8, 142.2, 142.1, 137.4, 136.8, 136.2, 134.9, 134.8, 134.6, 133.3, 133.0,131.8, 130.33, 130.30, 124.9, 114.9, 111.2 (d, J = 22.3 Hz), 106.16 (d, J =26.8 Hz). HRMS (ESI): For C 19 H 10 Cl2FN2O6S2, [MH] - Calculated value: 514.9347, measured value: 514.9360. HPLC purity: 98.91%.

[0175] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-fluorobenzoic acid (5f) Yellow solid (32 mg, 62%). mp 203 – 205 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 8.48(d, J = 1.3 Hz, 1H), 7.98 (d, J = 1.4 Hz, 1H), 7.82 (d, J = 8.2 Hz, 2H), 7.62(dd, J = 8.3, 1.6 Hz, 1H), 7.56 (dd,J = 9.2, 3.0 Hz, 1H), 7.40 (dd, J = 9.0, 4.8 Hz, 1H), 7.27 (td, J = 8.6, 2.9 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H). 13 C NMR (101 MHz, DMSO-) d 6) δ 168.1, 157.5 (d, J = 240.4 Hz), 144.4, 142.3, 139.1,138.5, 137.4, 136.1, 134.4, 133.3, 132.9, 131.8, 130.6, 130.1, 124.4, 122.9,121.8 (d, J = 6.4 Hz), 120.5 (d, J = 22.6 Hz), 117.2 (d, J = 23.3 Hz). HRMS(ESI): For C 19 H 10 Cl2FN2O6S2, [MH] - Calculated value: 514.9347, measured value: 514.9355. HPLC purity: 98.75%.

[0176] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-(trifluoromethyl)benzoic acid (5g) Yellow solid (40 mg, 71%). mp 124 – 126 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ14.56 (s, 1H), 8.51 (d, J = 1.4 Hz, 1H), 8.04 (d, J = 8.1 Hz, 1H), 7.98 (d, J = 1.5 Hz, 1H), 7.86 (dd, J = 8.6, 1.4 Hz, 1H), 7.82 (d, J = 8.3 Hz, 1H), 7.69(s, 1H), 7.62 (dd, J = 8.3, 1.5 Hz, 1H), 7.31 (d,J = 8.0 Hz, 1H), 7.11 (d, J = 8.6 Hz, 1H). 13 C NMR (100 MHz, DMSO- d 6) δ 168.1, 144.4, 143.7, 142.3, 139.2,137.4, 136.1, 134.4, 133.3, 133.2 (q, J = 31.6 Hz), 132.9, 132.8, 131.7,130.6, 130.3, 124.2, 123.94 (q, J = 271.2 Hz), 123.7, 118.2 (d, J = 3.4 Hz), 115.7 (d, J = 3.9 Hz). HRMS (ESI): For C 20 H 10 Cl2F3N2O6S2, [MH] - Calculated value: 564.9315, measured value: 564.9323. HPLC purity: 99.48%.

[0177] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-methoxybenzoic acid (5h) Yellow solid (37 mg, 69%). mp 213 – 215 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ11.96 – 11.51 (m, 1H), 8.58 (d, J = 1.9 Hz, 1H), 7.99 (d, J = 1.3 Hz, 1H), 7.94 (dd, J = 8.7, 1.9 Hz, 1H), 7.85 (t, J = 8.5 Hz, 2H), 7.63 (dd, J = 8.3, 1.7 Hz, 1H), 7.12 (d, J = 8.6 Hz, 1H), 6.98 (d, J = 2.2 Hz, 1H), 6.71 (dd, J = 8.9, 2.2 Hz, 1H), 3.79 (s, 3H). 13C NMR (100 MHz, DMSO- d 6) δ 169.9, 164.1, 144.5, 143.9, 141.8, 137.5, 136.7, 136.2, 134.6, 134.1, 133.4, 133.0, 131.9, 130.4, 130.3, 124.9, 110.0, 109.9, 103.8, 56.1. HRMS (ESI): For C 20 H 13 Cl2N2O7S2,[MH] - Calculated value: 526.9547, measured value: 526.9572. HPLC purity: 95.43%.

[0178] 2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4,5-dimethoxybenzoic acid (5i) Yellow solid (34 mg, 61%). mp 273 – 275 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 11.24 (s, 1H), 8.51 (d, J = 1.5 Hz, 1H), 7.98 (d, J = 1.5 Hz, 1H), 7.82 (dd, J = 11.2, 4.8 Hz, 2H), 7.62 (dd, J = 8.3, 1.7 Hz, 1H), 7.31 (s, 1H), 7.08 (t, J = 4.2 Hz, 2H), 3.82 (s, 3H), 3.72 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6) δ169.5, 153.8, 145.4, 144.5, 143.6, 137.4, 136.6, 136.1, 134.6, 134.3, 133.3, 133.0, 132.0, 130.4, 130.2, 124.8, 113.3, 110.6, 104.0, 56.2, 56.1. HRMS(ESI): For C 21 H 15 Cl2N2O8S2, [MH] -Calculated value: 556.9652, measured value: 556.9663. HPLC purity: 95.17%.

[0179] 4-Bromo-2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)benzoic acid (5j) Yellow solid (43 mg, 75%). mp 253 – 255 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 11.49 (s, 1H), 8.56 (d, J = 1.9 Hz, 1H), 7.99 (d, J = 1.9 Hz, 1H), 7.90 (dd, J = 8.7, 2.0 Hz, 1H), 7.82 (dd, J = 11.5, 8.4 Hz, 2H), 7.67 – 7.57 (m, 2H),7.38 (dd, J = 8.5, 1.7 Hz, 1H), 7.12 (d, J = 8.7 Hz, 1H). 13 C NMR (101 MHz, DMSO- d 6) δ 169.1, 144.6, 143.9, 140.5, 137.4, 136.7, 136.1, 134.6, 133.7, 133.4, 133.0, 131.8, 130.4, 130.3, 127.9, 127.5, 124.8, 122.5, 118.3. HRMS(ESI): For C 19 H 10 BrCl2N2O6S2, [MH] - Calculated value: 574.8546; Measured value: 574.8544. HPLC purity: 99.55%.

[0180] 1-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)cyclopropane-1-carboxylic acid (5k) Yellow solid (32 mg, 70%). mp 166 – 168 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 8.50(d, J = 1.9 Hz, 1H), 8.02 (d, J= 2.0 Hz, 1H), 7.90 – 7.84 (m, 2H), 7.66 (dd, J = 8.3, 2.1 Hz, 1H), 7.15 (d, J = 8.6 Hz, 1H), 1.29 – 1.21 (m, 2H), 1.13(dd, J = 7.5, 4.4 Hz, 2H). 13 C NMR (100 MHz, DMSO- d 6) δ 173.5, 144.4, 141.8, 140.3, 137.3, 136.0, 134.4, 133.3, 132.9, 132.2, 130.8, 129.7, 124.6, 35.6, 16.3. HRMS (ESI): For C 16 H 11 Cl2N2O6S2, [MH] - Calculated value: 460.9441, measured value: 460.9451. HPLC purity: 93.60%.

[0181] ((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonyl)-L-valine (5l) Yellow solid (35 mg, 73%). mp 147 – 149 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 8.55(d, J = 1.9 Hz, 1H), 8.33 (s, 1H), 8.01 (d, J = 1.9 Hz, 1H), 7.94 – 7.88 (m,1H), 7.88 – 7.84 (m, 1H), 7.65 (dd, J = 8.3, 2.0 Hz, 1H), 7.15 (d, J = 8.6Hz, 1H), 3.54 (d, J = 5.6 Hz, 1H), 1.98 (dq, J = 13.2, 6.6 Hz, 1H), 0.84 (d, J = 6.8 Hz, 3H), 0.81 (d, J = 6.8 Hz, 3H). 13 C NMR (100 MHz, DMSO-d 6) δ 172.3,144.4, 141.9, 139.4, 137.2, 136.0, 134.4, 133.3, 132.9, 132.2, 130.8, 129.9,124.6, 62.0, 30.7, 19.5, 18.2. HRMS (ESI): For C 17 H 15 Cl2 N2O6S2, [MH - Calculated value: 476.9754, Measured value: 476.9759. HPLC purity: 100.00%.

[0182] ((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonyl)-L-phenylalanine (5m) Yellow solid (41 mg, 77%). mp 183 – 185 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 8.22(s, 1H), 8.01 (s, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.64 (dd, J = 12.0, 9.3 Hz,2H), 7.07 (s, 5H), 6.93 (d, J = 8.6 Hz, 1H), 3.80 (dd, J = 8.9, 4.0 Hz, 1H), 2.97 (dd, J = 13.6, 4.0 Hz, 1H), 2.71 (dd, J = 13.5, 9.6 Hz, 1H). 13 C NMR (100MHz, DMSO- d 6) δ 172.5, 144.2, 141.7, 139.3, 137.9, 137.4, 136.1, 134.4, 133.3, 132.9, 131.6, 130.7, 129.8, 129.5, 128.2, 126.3, 124.1, 58.6, 38.3. HRMS (ESI): For C 21 H 15 Cl2N2O6S2, [MH] - Calculated value: 524.9754, measured value: 524.9751. HPLC purity: 99.30%.

[0183] ((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonyl)-L-alanine(5n) Yellow solid (34 mg, 76%). mp 133 – 135 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 12.69 (s, 1H), 8.55 (d, J = 1.8 Hz, 2H), 8.02 (d, J = 1.8 Hz, 1H), 7.97 –7.76 (m, 2H), 7.66 (dd, J = 8.2, 2.0 Hz, 1H), 7.16 (d, J = 8.6 Hz, 1H), 3.83(d, J = 6.4 Hz, 1H), 1.20 (d, J = 7.1 Hz, 3H). 13 C NMR (100 MHz, DMSO- d 6) δ173.4, 144.5, 142.0, 139.5, 137.3, 136.0, 134.4, 133.3, 132.9, 132.0, 130.8, 130.0, 124.5, 51.8, 18.9. HRMS (ESI): For C 15 H 11 Cl2N2O6S2, [MH] - Calculated value: 448.9441, measured value: 448.9447. HPLC purity: 97.82%.

[0184] General procedure for synthesizing 6a–6k A solution of 4c–4f (0.1 mmol) in DMF was added with 0.15 mmol of brominated material and 27 mg (0.2 mmol) of K₂CO₃, and the mixture was heated to 80 °C overnight. After cooling to ambient temperature, water was added. The mixture was extracted with ethyl acetate. The organic layer was washed successively with water and a saturated aqueous solution of NaCl, dried over Na₂SO₄, and concentrated. The residue was purified by silica gel column chromatography to give the title compound.

[0185] 2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (6a) Yellow solid (50 mg, 86%). 1 H NMR (400 MHz, CDCl3) δ 8.50 (d, J = 1.9 Hz, 1H), 7.70 (d, J = 1.7 Hz, 2H), 7.64 – 7.57 (m, 2H), 7.43 (d, J = 2.0 Hz, 1H),7.29 (s, 1H), 7.11 (d, J = 8.1 Hz, 1H), 6.90 (d, J = 8.6 Hz, 1H), 3.72 (s,3H), 3.68 (s, 1H), 3.40 (s, 1H), 2.40 (s, 3H), 1.04 – 0.93 (m, 1H), 0.43 (d, J = 7.8 Hz, 2H), 0.07 (d, J = 24.7 Hz, 2H).

[0186] 2-((N-benzyl-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (6b) Yellow solid (50 mg, 81%). 1 H NMR (400 MHz, CDCl3) δ 8.42 (d, J = 1.9 Hz, 1H), 7.68 (dd, J = 14.0, 1.8 Hz, 2H), 7.61 (d, J = 8.3 Hz, 1H), 7.55 (dd, J =8.6, 1.9 Hz, 1H), 7.42 (dd, J = 8.3, 2.0 Hz, 1H), 7.21 – 7.28 (m, 5H), 7.16(dd, J = 8.1, 1.6 Hz, 1H), 6.88 (d, J = 8.6 Hz, 1H), 6.74 (d, J = 8.0 Hz,1H), 5.06 (s, 1H), 4.63 (s, 1H), 3.70 (s, 3H), 2.34 (s, 3H).

[0187] 2-((4-((3,4-dichlorophenyl)thio)-N-(4-methoxybenzyl)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (6c) Yellow solid (55 mg, 85%). 1 H NMR (400 MHz, CDCl3) δ 8.41 (d, J = 1.7 Hz, 1H), 7.70 (d, J = 1.8 Hz, 1H), 7.66 (d, J = 1.2 Hz, 1H), 7.60 (d, J = 8.3 Hz, 1H), 7.54 (dd, J = 8.6, 1.7 Hz, 1H), 7.42 (dd, J = 8.3, 1.9 Hz, 1H), 7.15(dd, J = 13.8, 5.0 Hz, 3H), 6.87 (d, J = 8.5 Hz, 1H), 6.76 (d, J = 8.5 Hz, 2H), 6.70 (d, J = 8.0 Hz, 1H), 5.03 (d, J = 14.3 Hz, 1H), 4.55 (d, J = 14.2Hz, 1H), 3.77 (s, 3H), 3.71 (s, 3H), 2.34 (s, 3H).

[0188] 2-((N-(3-chlorobenzyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (6d) Yellow solid (50 mg, 74%). 1 H NMR (400 MHz, CDCl3) δ 8.42 (d, J = 1.9 Hz, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.67 (d, J = 1.6 Hz, 1H), 7.61 (d, J = 8.3 Hz, 1H), 7.56 (dd, J = 8.5, 2.0 Hz, 1H), 7.43 (dd, J= 8.3, 2.0 Hz, 1H), 7.26 –7.08 (m, 5H), 6.90 (d, J = 8.6 Hz, 1H), 6.78 (d, J = 8.1 Hz, 1H), 5.02 (s, 1H), 4.62 (s, 1H), 3.70 (s, 3H), 2.36 (s, 3H).

[0189] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamido)-5-methylbenzoate (6e) Yellow solid (58 mg, 82%). 1 H NMR (400 MHz, CDCl3) δ 8.47 (d, J = 1.9 Hz,1H), 7.69 (s, 2H), 7.65 (d, J = 8.1 Hz, 2H), 7.63 – 7.60 (m, 1H), 7.60 – 7.58(m, 1H), 7.41 (dd, J = 8.3, 2.0 Hz, 1H), 7.29 (s, 1H), 7.23 (dd, J = 8.1, 1.6Hz, 1H), 6.98 (d, J = 8.1 Hz, 1H), 6.91 (d, J = 8.6 Hz, 1H), 6.70 (s, 1H), 5.18 (d, J = 15.8 Hz, 1H), 4.75 (d, J = 15.3 Hz, 1H), 3.71 (s, 3H), 2.41 (s, 3H), 2.36 (s, 3H).

[0190] 2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-fluorobenzoate (6f) Yellow solid (40 mg, 68%). 1 H NMR (400 MHz, CDCl3) δ 8.53 (d, J = 1.9 Hz, 1H), 7.95 (dd, J= 8.8, 6.4 Hz, 1H), 7.70 (d, J = 1.9 Hz, 1H), 7.63 – 7.57(m, 2H), 7.43 (dd, J = 8.3, 2.0 Hz, 1H), 7.19 – 7.13 (m, 1H), 7.01 (dd, J =9.0, 2.5 Hz, 1H), 6.93 (d, J = 8.6 Hz, 1H), 3.72 (s, 4H), 3.39 (s, 1H), 1.04– 0.94 (m, 1H), 0.46 (d, J = 7.9 Hz, 2H), 0.08 (d, J = 27.8 Hz, 2H).

[0191] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamide)-4-fluorobenzoate (6g) Yellow solid (50 mg, 71%). 1 H NMR (400 MHz, CDCl3) δ 8.50 (d, J = 1.9 Hz, 1H), 7.95 (dd, J = 8.8, 6.3 Hz, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.66 (d, J =8.1 Hz, 2H), 7.60 (dd, J = 8.5, 2.2 Hz, 2H), 7.41 (dd, J = 8.3, 2.0 Hz, 1H), 7.28 (d, J = 8.5 Hz, 2H), 7.14 (ddd, J = 8.9, 7.5, 2.6 Hz, 1H), 6.93 (dd, J =8.7, 1.4 Hz, 2H), 6.68 (s, 1H), 5.17 (s, 1H), 4.78 (s, 1H), 3.72 (s, 3H), 2.42 (s, 3H).

[0192] 4-Chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamide)methyl benzoate (6h) Yellow solid (55 mg, 76%). 1 H NMR (400 MHz, CDCl3) δ 8.49 (d, J = 2.0 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.70 (d, J = 1.9 Hz, 1H), 7.66 (d, J = 8.1 Hz,2H), 7.62 – 7.56 (m, 2H), 7.42 (dd, J = 8.3, 2.0 Hz, 2H), 7.29 (d, J = 8.1Hz, 2H), 7.16 (d, J = 2.0 Hz, 1H), 6.93 (d, J = 8.6 Hz, 1H), 6.68 (s, 1H), 5.13 (s, 1H), 4.76 (s, 1H), 3.74 (s, 3H), 2.42 (s, 3H).

[0193] 4-Chloro-2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)methyl benzoate (6i) Yellow solid (42 mg, 70%). 1 H NMR (400 MHz, CDCl3) δ 8.53 (d, J = 1.9 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.64 – 7.56 (m, 2H), 7.46 – 7.40 (m, 2H), 7.23 (d, J = 2.0 Hz, 1H), 6.93 (d, J = 8.6 Hz, 1H), 3.74(s, 3H), 3.67 (s, 1H), 3.41 (s, 1H), 1.05 – 0.93 (m, 1H), 0.47 (d, J= 7.8Hz, 2H), 0.08 (d, J = 27.4 Hz, 2H).

[0194] 2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-fluorobenzoate (6j) Yellow solid (42 mg, 70%). 1 H NMR (400 MHz, CDCl3) δ 8.52 (d, J = 1.9 Hz, 1H), 7.70 (d, J = 2.0 Hz, 1H), 7.66 – 7.53 (m, 3H), 7.42 (dd, J = 8.3, 2.0Hz, 1H), 7.26 – 7.13 (m, 2H), 6.92 (d, J = 8.6 Hz, 1H), 3.77 – 3.68 (m, 4H), 3.38 (dd, J = 14.0, 7.3 Hz, 1H), 0.97 (qd, J = 7.5, 3.7 Hz, 1H), 0.43 (t, J =9.6 Hz, 2H), 0.07 (d, J = 35.8 Hz, 2H).

[0195] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamido)-5-fluorobenzoate (6k) Yellow solid (65 mg, 93%). 1 H NMR (400 MHz, CDCl3) δ 8.49 (d, J = 2.0 Hz, 1H), 7.69 (d, J = 2.0 Hz, 1H), 7.65 (d, J = 8.1 Hz, 2H), 7.63 – 7.55 (m, 3H), 7.41 (dd, J = 8.3, 2.0 Hz, 1H), 7.28 (d, J = 8.0 Hz, 2H), 7.13 (dd, J = 6.0, 1.5 Hz, 2H), 6.92 (d,J = 8.7 Hz, 1H), 6.67 (s, 1H), 5.21 (d, J = 16.1 Hz, 1H), 4.73 (d, J = 16.3 Hz, 1H), 3.74 (s, 3H), 2.42 (s, 3H).

[0196] General procedure for synthesizing 7a-7k The methyl ester (0.1 mmol) of the title compound (6a – 6k) was used at 50 °C in 1 M NaOH (1 mmol) in dioxane. 1) Hydrolyze overnight. The mixture was then diluted with a small amount of water and washed twice with DCM. The aqueous solution was acidified by adding 2 M HCl. The precipitate was collected by filtration and washed with water to provide the title compounds (7a – 7k).

[0197] 2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (7a) Yellow solid (35 mg, 62%). mp 207 – 209 ℃. 1 H NMR (400 MHz, CDCl3) δ 8.55(d, J = 1.8 Hz, 1H), 7.84 (d, J = 1.3 Hz, 1H), 7.66 (d, J = 1.9 Hz, 1H), 7.61– 7.50 (m, 2H), 7.42 – 7.32 (m, 2H), 7.14 (d, J = 8.1 Hz, 1H), 6.91 (d, J =8.6 Hz, 1H), 3.70 (dd, J = 14.1, 6.7 Hz, 1H), 3.41 (dd, J = 14.1, 7.4 Hz,1H), 2.44 (s, 3H), 1.04 – 0.93 (m, 1H), 0.51 – 0.40 (m, 2H), 0.10 (d, J =30.6 Hz, 2H). 13C10 NMR (100 MHz, CDCl3) δ 169.5, 144.3, 143.4, 139.7, 137.6, 137.1, 135.7, 135.4, 134.8, 134.5, 134.2, 133.2, 132.3, 131.6, 129.7, 129.6, 128.4, 125.2, 57.5, 21.1, 10.4, 4.4, 3.8. HRMS (ESI): for C10 NMR... 24 H 19 Cl2N2O6S2, [MH] - Calculated value: 565.0067, measured value: 565.0053. HPLC purity: 98.98%.

[0198] 2-((N-benzyl-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (7b) Yellow solid (32 mg, 53%). mp 145 – 147 ℃. 1 H NMR (400 MHz, CDCl3) δ 8.52(d, J = 1.6 Hz, 1H), 7.81 (s, 1H), 7.65 (d, J = 1.9 Hz, 1H), 7.55 (d, J = 8.2Hz, 1H), 7.51 (dd, J = 8.6, 1.7 Hz, 1H), 7.37 (dd, J = 8.2, 2.0 Hz, 1H), 7.28– 7.20 (m, 6H), 6.88 (d, J = 8.6 Hz, 1H), 6.78 (d, J = 8.0 Hz, 1H), 5.13 (d, J = 14.5 Hz, 1H), 4.62 (d, J = 14.5 Hz, 1H), 2.39 (s, 3H). 13C10 NMR (100 MHz, CDCl3) δ 169.7, 144.4, 143.3, 139.6, 137.9, 137.1, 135.7, 135.5, 135.0, 134.8, 134.5, 134.3, 133.3, 133.0, 132.3, 131.6, 129.8, 129.5, 128.6, 128.4, 128.2, 125.3, 56.4, 21.0. HRMS (ESI): for C10 NMR... 27 H 19 Cl2N2O6S2, [MH] - Calculated value: 601.0067, measured value: 601.0046. HPLC purity: 98.42%.

[0199] 2-((4-((3,4-dichlorophenyl)thio)-N-(4-methoxybenzyl)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (7c) Yellow solid (42 mg, 66%). mp 143 – 145 ℃. 1 H NMR (400 MHz, CDCl3) δ 8.51(d, J = 1.7 Hz, 1H), 7.82 (s, 1H), 7.64 (d, J = 1.9 Hz, 1H), 7.54 (d, J = 8.3Hz, 1H), 7.50 (dd, J = 8.5, 1.7 Hz, 1H), 7.36 (dd, J = 8.3, 1.9 Hz, 1H), 7.23(d, J = 8.0 Hz, 1H), 7.16 (d, J = 8.4 Hz, 2H), 6.87 (d, J = 8.6 Hz, 1H), 6.76(t, J = 8.8 Hz, 3H), 5.08 (d, J = 14.6 Hz, 1H), 4.55 (d, J = 14.6 Hz, 1H), 3.77 (s, 3H), 2.40 (s, 3H). 13C10 NMR (100 MHz, CDCl3) δ 170.2, 159.4, 144.3, 143.2, 139.6, 138.1, 137.1, 135.6, 135.0, 134.8, 134.5, 134.3, 133.3, 133.1, 132.3, 131.6, 130.9, 129.8, 129.1, 128.4, 127.5, 125.2, 113.8, 55.9, 55.2, 21.1. HRMS (ESI): for C10 NMR... 28 H 21 Cl2N2O7S2, [MH] - Calculated value: 631.0173, measured value: 631.0180. HPLC purity: 99.12%.

[0200] 2-((N-(3-chlorobenzyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (7d) Yellow solid (42 mg, 66%). mp 129 – 131 °C. 1 H NMR (400 MHz, CDCl3) δ 8.52(d, J = 1.9 Hz, 1H), 7.83 (d, J = 1.4 Hz, 1H), 7.66 (d, J = 1.9 Hz, 1H), 7.56(d, J = 8.3 Hz, 1H), 7.51 (dd, J = 8.6, 1.9 Hz, 1H), 7.38 (dd, J = 8.3, 2.0Hz, 1H), 7.29 (d, J = 1.3 Hz, 1H), 7.26 – 7.15 (m, 4H), 6.90 (d, J = 8.6 Hz, 1H), 6.83 (d, J = 8.1 Hz, 1H), 5.10 (d, J = 15.0 Hz, 1H), 4.58 (d, J = 15.0Hz, 1H), 2.41 (s, 3H). 13C10 NMR (100 MHz, CDCl3) δ 169.6, 144.3, 143.6, 139.9, 137.73, 137.71, 137.1, 135.7, 134.9, 134.8, 134.5, 134.4, 133.5, 133.0, 132.3, 131.6, 129.9, 129.7, 129.4, 128.7, 128.4, 128.3, 127.6, 125.3, 55.8, 21.1. HRMS (ESI): for C10 NMR... 27 H 18 Cl3N2O6S2, [MH] - Calculated value: 634.9677, measured value: 634.9681. HPLC purity: 96.91%.

[0201] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamido)-5-methylbenzoic acid (7e) Yellow solid (40 mg, 58%). mp 191 – 193 ℃. 1 H NMR (400 MHz, CDCl3) δ 8.54(d, J = 1.8 Hz, 1H), 7.79 (d, J = 1.1 Hz, 1H), 7.65 (dd, J = 11.2, 5.0 Hz, 3H), 7.57 (td, J = 8.1, 3.9 Hz, 2H), 7.38 (dd, J = 8.3, 1.9 Hz, 1H), 7.30 –7.26 (m, 3H), 6.99 (d, J = 8.1 Hz, 1H), 6.93 (d, J = 8.6 Hz, 1H), 6.64 (s,1H), 5.08 (d, J = 16.2 Hz, 1H), 4.85 (d, J = 16.0 Hz, 1H), 2.40 (s, 6H). 13 CNMR (100 MHz, DMSO- d6) δ 169.9, 167.1, 161.2, 144.6, 143.1, 140.9, 139.6, 137.2, 135.9, 134.8, 134.4, 133.6, 133.3, 132.9, 132.4, 132.3, 132.0, 130.7, 130.3, 130.0, 126.0, 125.1, 124.4, 100.1, 47.7, 21.5, 20.9. HRMS (ESI): For C 31 H 22 Cl2N3O7S2, [MH] - Calculated value: 682.0282, measured value: 682.0290. HPLC purity: 97.92%.

[0202] 2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-fluorobenzoic acid (7f) Yellow solid (31 mg, 53%). mp 136–138 ℃. 1 H NMR (400 MHz, CDCl3) δ 8.59(d, J = 1.4 Hz, 1H), 8.13 – 8.02 (m, 1H), 7.66 (d, J = 1.8 Hz, 1H), 7.57 (t, J = 9.1 Hz, 2H), 7.39 (dd, J = 8.3, 1.8 Hz, 1H), 7.26 – 7.18 (m, 1H), 7.05(d, J = 8.4 Hz, 1H), 6.92 (d, J = 8.6 Hz, 1H), 3.72 (s, 1H), 3.42 (s, 1H), 0.99 (s, 1H), 0.48 (d, J = 7.9 Hz, 2H), 0.10 (d, J = 39.8 Hz, 2H). 13 C NMR (100MHz, CDCl3) δ 168.3, 164.9 (d, J = 257.3 Hz), 144.3, 143.8, 140.7 (d, J=10.9 Hz), 137.4, 137.2, 135.8, 134.85, 134.80, 134.76, 134.6, 132.4, 131.5,129.6, 128.5, 125.2, 120.4 (d, J = 22.1 Hz), 116.5 (d, J = 20.9 Hz), 57.4, 10.3, 4.5, 3.7. HRMS (ESI): for C 23 H 16 Cl2 FN2O6S2, [MH - Calculated value: 568.9816, measured value: 568.9799. HPLC purity: 97.97%.

[0203] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamide)-4-fluorobenzoic acid (7g) Yellow solid (41 mg, 60%). mp 153 – 155 ℃. 1 H NMR (400 MHz, DMSO- d 6 ) δ13.15 (s, 1H), 8.35 (d, J = 1.9 Hz, 1H), 8.00 – 7.89 (m, 2H), 7.84 (d, J =8.3 Hz, 1H), 7.69 (d, J = 8.1 Hz, 2H), 7.64 (ddd, J = 16.7, 8.6, 2.0 Hz, 2H),7.40 (dd, J = 8.3, 2.3 Hz, 1H), 7.36 (d, J = 8.0 Hz, 2H), 7.24 (dd, J = 9.5, 2.3 Hz, 1H), 7.10 (d, J = 8.6 Hz, 1H), 6.93 (s, 1H), 5.11 (d, J = 14.5 Hz, 1H), 4.83 (d, J = 14.6 Hz, 1H), 2.38 (s, 3H). 13 C NMR (100 MHz, DMSO-d 6) δ169.9, 166.1, 163.8 (d, J = 250 Hz), 161.0, 144.7, 143.3, 140.9, 137.2,136.6, 135.9, 134.5, 134.3, 134.2 ( d , = 10 Hz), 133.4, 132.9, 132.4, 130.7,130.3, 130.0, 129.1, 126.0, 125.3, 124.4, 119.4 (d, J = 23 Hz), 116.9 (d, J =21 Hz), 100.1, 47.6, 21.5. HRMS (ESI): for C 30 H 19 Cl2FN3O7S2, [MH] - Calculated value: 686.0031, measured value: 686.0015. HPLC purity: 92.97%.

[0204] 4-Chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamide)benzoic acid (7h) Yellow solid (47 mg, 66%). mp 241 – 243 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 8.38(s, 1H), 7.99 (s, 1H), 7.81 (t, J = 7.5 Hz, 2H), 7.72 (d, J = 7.8 Hz, 2H),7.69 – 7.56 (m, 2H), 7.35 (d, J = 7.4 Hz, 3H), 7.18 (s, 1H), 7.07 (d, J = 8.6Hz, 1H), 6.98 (s, 1H), 5.07 (s, 2H), 2.38 (s, 3H). 13 C NMR (100 MHz, DMSO- d6) δ169.8, 167.6, 161.9, 144.6, 142.4, 140.8, 137.5, 137.0, 136.7, 135.8, 134.2, 133.3, 133.0, 132.8, 132.7, 131.9, 131.1, 130.2, 129.6, 128.9, 126.0, 125.8, 124.5, 100.0, 47.6, 21.5. HRMS (ESI): For C 30 H 19 Cl3N3O7S2, [MH] - Calculated value: 701.9735, measured value: 701.9742. HPLC purity: 99.04%.

[0205] 4-Chloro-2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)benzoic acid (7i) Yellow solid (36 mg, 61%). mp 115 – 117 ℃. 1 H NMR (400 MHz, CDCl3) δ 8.58(d, J = 1.6 Hz, 1H), 7.99 (d, J = 8.5 Hz, 1H), 7.67 (d, J = 1.8 Hz, 1H), 7.59(d, J = 8.3 Hz, 1H), 7.54 (dd, J = 8.6, 1.3 Hz, 1H), 7.49 (d, J = 8.3 Hz, 1H), 7.40 (dd, J = 8.2, 1.8 Hz, 1H), 7.23 (s, 1H), 6.93 (d, J = 8.6 Hz, 1H), 3.68 (dd, J = 13.8, 6.9 Hz, 1H), 3.41 (dd, J = 13.9, 6.9 Hz, 1H), 1.06 – 0.89(m, 1H), 0.48 (d, J = 7.8 Hz, 2H), 0.21 – 0.01 (m, 2H). 13C10 NMR (100 MHz, CDCl3) δ 168.1, 150.7, 144.4, 143.9, 139.4, 139.3, 137.1, 135.8, 134.8, 134.6, 133.6, 132.7, 132.4, 131.6, 129.6, 129.5, 128.5, 125.2, 57.4, 10.3, 4.4, 3.9. HRMS (ESI): for C10 NMR... 23 H 16 Cl3N2O6S2, [MH] - Calculated value: 584.9521, measured value: 584.9504. HPLC purity: 96.65%.

[0206] 2-((N-(cyclopropylmethyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-fluorobenzoic acid (7j) Yellow solid (27 mg, 52%). mp 69 – 71 °C. 1 H NMR (400 MHz, CDCl3) δ 8.56 (d, J = 1.8 Hz, 1H), 7.72 (d, J = 8.5 Hz, 1H), 7.67 (d, J = 1.9 Hz, 1H), 7.59 (d, J = 8.3 Hz, 1H), 7.56 (dd, J = 8.7, 1.9 Hz, 1H), 7.40 (dd, J = 8.3, 1.9 Hz, 1H), 7.28 (dd, J = 6.1, 1.4 Hz, 2H), 6.92 (d, J = 8.6 Hz, 1H), 3.73 (dd, J =14.2, 6.9 Hz, 1H), 3.40 (dd, J = 14.2, 7.5 Hz, 1H), 0.98 (dt, J = 7.5, 4.9Hz, 1H), 0.46 (t, J = 9.5 Hz, 2H), 0.19 – 0.01 (m, 2H). 13 C NMR (100 MHz, CDCl3) δ 168.0, 161.9 (d,J = 250.6 Hz), 144.4, 143.7, 137.5, 137.2, 135.8,134.8, 134.6, 134.5, 134.1 (d, J = 4.0 Hz), 132.4, 132.0 (d, J = 8.0 Hz),131.5, 129.6, 128.5, 125.2, 120.5 (d, J = 23.0 Hz), 119.5 (d, J = 25.0 Hz), 57.5, 10.3, 4.4, 3.8. HRMS (ESI): For C 23 H 16 Cl2FN2O6S2, [MH] - Calculated value: 568.9816, measured value: 568.9819. HPLC purity: 100.00%.

[0207] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-((5-(p-tolyl)isoxazo-3-yl)methyl)phenyl)sulfonamido)-5-fluorobenzoic acid (7k) Yellow solid (36 mg, 53%). mp 285 – 287. 1 H NMR (400 MHz, DMSO- d 6) δ 8.37(d, J = 1.9 Hz, 1H), 8.01 (d, J = 1.8 Hz, 1H), 7.86 – 7.76 (m, 2H), 7.72 (d, J = 8.1 Hz, 2H), 7.66 (dd, J = 8.3, 1.8 Hz, 1H), 7.35 (d, J = 8.0 Hz, 2H), 7.27 (d, J = 7.7 Hz, 1H), 7.15 – 7.00 (m, 3H), 6.92 (s, 1H), 5.07 (s, 2H), 2.38 (s, 3H). 13 C NMR (100 MHz, DMSO- d 6) δ 169.7, 167.1, 161.8 (d, J= 245.0Hz), 161.9, 144.6, 142.3, 140.8, 137.8, 137.0, 135.8, 134.84, 134.76, 134.2,133.2, 132.9, 132.8, 131.5, 131.2, 130.2, 129.7, 126.0, 125.7, 124.5, 117.1(d, J = 22.9 Hz), 115.4 (d, J = 23.0 Hz), 100.0, 47.6, 21.5. HRMS (ESI): For C 30 H 19 Cl2FN3O7S2, [MH] - Calculated value: 686.0031, measured value: 686.0036. HPLC purity: 99.48%.

[0208] Synthesis of methyl 2-((N-(3-bromopropyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (8) 1,4-Dibromobutane (7.5 mmol) and K₂CO₃ (1.38 g, 10 mmol) were added to a solution of 4c (2.64 g, 5 mmol) in DMF / acetone at room temperature, and the mixture was heated to 60 °C overnight. After cooling to ambient temperature, water was added. The mixture was extracted with ethyl acetate. The organic layer was washed successively with water and a saturated aqueous solution of NaCl, dried over Na₂SO₄, and concentrated. The residue was purified by silica gel column chromatography to give the title compound as a yellow gel (2.33 g, 72%). 1 H NMR (400 MHz, CDCl3) δ 8.43 (d, J = 1.8 Hz, 1H), 7.70 (d, J = 1.9 Hz, 2H), 7.61(d, J = 8.3 Hz, 1H), 7.55 (dd, J = 8.6, 1.9 Hz, 1H), 7.42 (dd, J = 8.3, 1.9Hz, 1H), 7.30 (dd, J = 8.0, 1.5 Hz, 1H), 6.98 (d, J = 8.1 Hz, 1H), 6.91 (d, J= 8.6 Hz, 1H), 3.74 (s, 3H), 3.65 (dd, J = 16.0, 6.9 Hz, 2H), 3.38 (d, J =3.8 Hz, 2H), 2.40 (s, 3H), 1.99 – 1.86 (m, 2H), 1.71 (dt, J = 14.7, 7.3 Hz, 2H).

[0209] General procedure for synthesizing compounds 9a–9d Amine (0.3 mmol) and K₂CO₃ (55 mg, 0.4 mmol) were added to a solution of 8 (133 mg, 0.2 mmol) in DMF / acetone at room temperature, and the mixture was heated to 40 °C overnight. After cooling to ambient temperature, water was added. The mixture was extracted with ethyl acetate. The organic layer was washed successively with water and a saturated aqueous solution of NaCl, dried over Na₂SO₄, and concentrated. The residue was purified by silica gel column chromatography to give the compound. 9a – 9d .

[0210] 2-((4-((3,4-dichlorophenyl)thio)-N-(4-(diethylamino)butyl)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (9a) Yellow solid (120 mg, 92%). 1 H NMR (400 MHz, CDCl3) δ 8.39 (d, J = 1.8 Hz, 1H), 7.71 (d, J = 1.9 Hz, 1H), 7.68 (s, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.52 (dd, J = 8.6, 1.9 Hz, 1H), 7.44 (dd, J = 8.3, 2.0 Hz, 1H), 7.30 (d, J = 1.7Hz, 1H), 6.91 (dd, J = 10.6, 8.5 Hz, 2H), 3.77 (s, 3H), 3.71 – 3.55 (m, 2H), 2.97 (d, J= 5.9 Hz, 4H), 2.84 (s, 2H), 2.40 (s, 3H), 1.71 – 1.56 (m, 4H), 1.33 – 1.28 (m, 6H).

[0211] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-(4-(pyrrolidine-1-yl)butyl)phenyl)sulfonamido)-5-methylbenzoate (9b) Yellow solid (70 mg, 54%). 1 H NMR (400 MHz, CDCl3) δ 8.42 (d, J = 1.8 Hz, 1H), 7.70 (dd, J = 5.6, 1.7 Hz, 2H), 7.61 (d, J = 8.3 Hz, 1H), 7.55 (dd, J =8.6, 1.9 Hz, 1H), 7.43 (dd, J = 8.3, 2.0 Hz, 1H), 7.29 (dd, J = 8.2, 1.6 Hz, 1H), 6.93 (dd, J = 12.1, 8.4 Hz, 2H), 3.75 (s, 3H), 3.63 (t, J = 7.1 Hz, 2H), 2.71 (d, J = 48.5 Hz, 6H), 2.40 (s, 3H), 1.90 (s, 4H), 1.71 (s, 2H), 1.66 –1.59 (m, 2H).

[0212] 2-((4-((3,4-dichlorophenyl)thio)-N-(4-(3,3-difluoropyrrolidone-1-yl)butyl)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (9c) Yellow solid (90 mg, 65%). 1 H NMR (400 MHz, CDCl3) δ 8.43 (d, J = 1.6 Hz, 1H), 7.70 (d, J = 1.8 Hz, 2H), 7.61 (d, J = 8.3 Hz, 1H), 7.55 (d, J = 8.4 Hz, 1H), 7.42 (dd,J = 8.3, 2.0 Hz, 1H), 7.30 (dd, J = 8.1, 1.5 Hz, 1H), 6.98 (d, J = 8.1 Hz, 1H), 6.91 (d, J = 8.6 Hz, 1H), 4.05 (d, J = 5.6 Hz, 2H), 3.73 (s,3H), 3.69 – 3.47 (m, 6H), 2.40 (s, 3H), 2.33 (td, J = 13.3, 6.8 Hz, 2H), 1.66(s, 4H).

[0213] 2-((4-((3,4-dichlorophenyl)thio)-N-(4-morpholinobutyl)-3-nitrophenyl)sulfonamido)-5-methylbenzoate (9d) Yellow solid (104 mg, 78%). 1 H NMR (400 MHz, CDCl3) δ 8.45 (d, J = 1.6 Hz, 1H), 7.70 (d, J = 1.5 Hz, 2H), 7.61 (d, J = 8.2 Hz, 1H), 7.55 (dd, J = 8.7, 1.8 Hz, 1H), 7.42 (dd, J = 8.3, 1.9 Hz, 1H), 7.30 (d, J = 8.0 Hz, 1H), 6.99(d, J = 8.1 Hz, 1H), 6.90 (d, J = 8.6 Hz, 1H), 3.74 (s, 3H), 3.69 – 3.58 (m,6H), 2.40 (s, 3H), 2.37 (s, 4H), 2.31 – 2.25 (m, 2H), 1.58 – 1.63 (m, 2H),1.47 (dt, J = 14.0, 7.1 Hz, 2H).

[0214] General procedure for synthesizing 10a–10d The methyl ester (0.1 mmol) of the title compound (9a–9d) was used at 50 °C in 1 M NaOH (1 mmol) in dioxane. 1) Hydrolyze overnight. The mixture was then diluted with a small amount of water and washed twice with DCM. The aqueous solution was acidified by adding 2 M HCl. The precipitate was collected by filtration and washed with water to provide title compounds 10a–10d.

[0215] 2-((4-((3,4-dichlorophenyl)thio)-N-(4-(diethylamino)butyl)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (10a) Yellow solid (15 mg, 23%). mp 196 – 198 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 8.32(d, J = 1.6 Hz, 1H), 8.01 (d, J = 1.9 Hz, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.67(td, J = 8.4, 1.6 Hz, 2H), 7.50 (s, 1H), 7.25 (d, J = 7.9 Hz, 1H), 7.10 (d, J = 8.6 Hz, 1H), 6.98 (d, J = 8.1 Hz, 1H), 3.60 – 3.50 (m, 2H), 2.82 (q, J =6.3 Hz, 4H), 2.71 (s, 2H), 2.32 (s, 3H), 1.55 – 1.40 (m, 4H), 1.07 (t, J =7.1 Hz, 6H). 13 C NMR (100 MHz, DMSO- d 6) δ 168.0, 144.1, 141.9, 138.0, 136.6, 135.4, 133.9, 133.2, 132.8, 132.4, 132.0, 131.1, 130.8, 130.2, 130.1, 129.24, 129.19, 124.7, 50.7, 50.6, 46.0, 45.9, 25.4, 21.3, 20.4, 9.2, 9.1. HRMS (ESI): For C 28H 30 Cl2N3O6S2, [MH] - Calculated value: 638.0959, measured value: 638.0968. HPLC purity: 93.37%.

[0216] 2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-(4-(pyrrolidine-1-yl)butyl)phenyl)sulfonamido)-5-methylbenzoic acid (10b) Yellow solid (52 mg, 41%). mp 221 – 223 ℃. 1 H NMR (400 MHz, CDCl3) δ 8.54(d, J = 1.8 Hz, 1H), 7.70 (d, J = 1.9 Hz, 1H), 7.64 – 7.56 (m, 3H), 7.44 (dd, J = 8.2, 2.0 Hz, 1H), 7.11 (d, J = 8.1 Hz, 1H), 6.93 (d, J = 8.6 Hz, 1H), 6.80 (d, J = 8.0 Hz, 1H), 3.64 (s, 2H), 3.23 (s, 6H), 3.00 – 2.94 (m, 2H), 2.34 (s, 3H), 2.04 (s, 4H), 1.66 – 1.52 (m, 2H). 13 C10 NMR (100 MHz, CDCl3) δ 170.9, 144.3, 143.0, 138.9, 137.5, 137.3, 137.1, 135.6, 134.9, 134.4, 133.2, 132.3, 132.1, 131.7, 130.9, 129.8, 129.0, 128.4, 125.5, 55.0, 53.5, 51.3, 25.7, 23.3, 23.0, 21.0. HRMS (ESI): for C10 NMR... 28 H 28 Cl2N3O6S2, [MH] - Calculated value: 636.0802, measured value: 636.0811. HPCL purity: 97.78%.

[0217] 2-((4-((3,4-dichlorophenyl)thio)-N-(4-(3,3-difluoropyrrolidone-1-yl)butyl)-3-nitrophenyl)sulfonamido)-5-methylbenzoic acid (10c) Yellow solid (54 mg, 40%). mp 98 – 100 ℃. 1H NMR (400 MHz, CDCl3) δ 8.50 (s, 1H), 7.83 (s, 1H), 7.66 (d, δ ) J = 1.5 Hz, 1H), 7.56 (d, J = 8.3 Hz, 1H), 7.51 (d, J = 8.4 Hz, 1H), 7.44 – 7.32 (m, 2H), 7.04 (d, J = 7.9 Hz, 1H), 6.91(d, J = 8.6 Hz, 1H), 4.06 (s, 2H), 3.75 – 3.49 (m, 6H), 2.43 (s, 3H), 2.32(dd, J = 12.6, 6.4 Hz, 2H), 1.67 (s, 4H). 13 C NMR (100 MHz, CDCl3) δ 169.8,154.7, 144.8, 143.5, 139.6, 137.2, 137.1, 135.7, 135.0, 134.9, 134.5, 134.2,133.2, 132.3, 131.6, 131.3, 129.9 (t, J = 248.6 Hz), 129.7, 128.4, 125.2,65.0, 52.7 (td, J = 32.2 Hz, 12.0 Hz), 51.9 (d, J = 9.1 Hz), 43.6, 33.7 (dt, J = 45.6 Hz, 24.3 Hz), 26.2, 25.2 (d, J = 8.6 Hz), 21.0. HRMS (ESI): For C 28 H 26 Cl2F2N3O6S2, [MH] - Calculated value: 672.0614, measured value: 672.0608. HPLC purity: 96.69%.

[0218] 4-Chloro-2-((4-((3,4-dichlorophenyl)thio)-N-(4-morpholinobutyl)-3-nitrophenyl)sulfonamide)benzoic acid (10d) Yellow solid (25 mg, 39%). mp 163 – 165 ℃. 1 H NMR (400 MHz, DMSO- d 6) δ 8.26(d, J = 1.8 Hz, 1H), 8.02 (d, J = 1.9 Hz, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.67 (dd, J = 8.3, 1.9 Hz, 1H), 7.65 – 7.57 (m, 2H), 7.35 (d, J = 6.7 Hz, 1H), 7.12 (d, J = 8.6 Hz, 1H), 7.03 (d, J = 8.1 Hz, 1H), 3.80 (s, 4H), 3.61 (t, J = 6.1 Hz, 4H), 2.96 (s, 4H), 2.35 (s, 3H), 1.69 (s, 2H), 1.46 (s, 2H). 13 C NMR (100 MHz, DMSO-) d 6) δ 167.4, 144.7, 142.8, 139.0, 137.2, 136.7, 136.0, 134.5, 133.4, 133.2, 133.1, 133.0, 132.4, 132.0, 130.8, 130.7, 130.0, 124.9, 63.8, 56.0, 51.5, 51.2, 25.7, 20.9, 20.6. HRMS (ESI): For C 28 H 28 Cl2N3O7S2, [MH] - Calculated value: 652.0751, measured value: 652.0759. HPLC purity: 99.12%.

[0219] Program for synthesizing compound C3-332 Scheme 4. Synthesis of C3-332

[0220] Synthesis of methyl 4-chloro-2-((4-chloro-3-nitrophenyl)sulfonamide)benzoate (4d)

[0221] 4-Chloro-3-nitrobenzenesulfonyl chloride 1 (3.0 mg, 16 mmol, 1 equivalent) was dissolved in DCM. Pyridine (9.76 mL, 120 mmol, 7.5 equivalent) was added to the reaction mixture and cooled to 0°C. Methyl 2-amino-4-chlorobenzoate dissolved in DCM was slowly added to the reaction mixture and stirred overnight at room temperature. Water was added to the reaction mixture and the aqueous phase was extracted with DCM. The combined organic phases were purified by column chromatography, providing 4d as a yellow solid, 2.569 g, 39% yield. 1 H NMR (400 MHz, DMSO) δ 10.69 (s, 1H), 8.49 (d, J = 1.9 Hz, 1H), 8.04 (dd, J = 8.5, 2.0 Hz, 1H), 8.00 (d, J = 8.4 Hz, 1H), 7.84 (d, J = 8.5 Hz, 1H), 7.44 (d, J =2.0 Hz, 1H), 7.40 (dd, J = 8.4, 2.1 Hz, 1H), 3.76 (s, 3H).

[0222] Synthesis of methyl 4-chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)benzoate (6d)

[0223] Intermediate 4d (2.56 g, 6.3 mmol, 1 equivalent) and sodium acetate (2.59 g, 32 mmol, 5 equivalent) were dissolved in EtOH. 3,4-Dichlorothiophenol (1.89 g, 11 mmol, 1.67 equivalent) was added to the reaction mixture and heated to reflux for 6 h. The reaction mixture was then cooled to room temperature. The precipitate was collected by filtration and washed successively with small amounts of EtOH and water. The filter cake was dried in an oven to provide 6d as a yellow solid, 2.89 g, 84% yield. 1 H NMR (400 MHz, DMSO)δ 10.66 (s, 1H), 8.52 (d, J = 2.1 Hz, 1H), 8.01 (d, J= 2.1 Hz, 1H), 7.88(dd, J = 8.6, 2.1 Hz, 1H), 7.86 (d, J = 8.3 Hz, 1H), 7.73 (d, J = 8.5 Hz, 1H), 7.65 (dd, J = 8.4, 2.1 Hz, 1H), 7.41 (d, J = 2.1 Hz, 1H), 7.24 (s, 1H), 7.14 (d, J = 8.6 Hz, 1H), 3.75 (s, 3H).

[0224] Synthesis of tert-butyl piperidine-1-carboxylate (11)

[0225] Intermediate 6d (300 mg, 0.55 mmol, 1 equivalent), potassium carbonate (302 mg, 2.19 mmol, 4 equivalents), and potassium iodide (9 mg, 0.055 mmol, 0.1 equivalents) were dissolved in DMF. Tert-butyl 4-(bromomethyl)piperidine-1-carboxylate (609 mg, 2.19 mmol, 4 equivalents) was added to the reaction mixture and heated to 100°C overnight. Water was added to the reaction mixture, and the aqueous phase was extracted with ethyl acetate. The combined organic phases were purified by column chromatography to provide 11 as a yellow solid, 127 mg, in 31% yield. 1 H NMR (400 MHz, DMSO) δ 8.20 (d, J = 2.1 Hz, 1H), 8.02 (d, J =2.1 Hz, 1H), 7.86 (dd, J = 15.5, 8.4 Hz, 2H), 7.67 (dd, J = 8.3, 2.1 Hz, 1H), 7.60 (dd, J = 8.5, 2.0 Hz, 1H), 7.56 (dd, J = 8.6, 2.1 Hz, 1H), 7.34 (d, J =2.0 Hz, 1H), 7.16 (d, J= 8.7 Hz, 1H), 3.76 (s, 3H), 3.48 (q, J = 6.7 Hz,2H), 2.96 – 2.55 (m, 4H), 1.95 (d, J = 13.2 Hz, 1H), 1.79 – 1.44 (m, 4H), 1.39 (s, 9H).

[0226] Synthesis of methyl 4-chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-(piperidin-4-ylmethyl)phenyl)sulfonamide)benzoate (12)

[0227] Intermediate 11 (120 mg, 0.16 mmol, 1 equivalent) was dissolved in 3 mL of DCM. 1 mL of TFA was slowly added to the reaction mixture at a DCM:TFA ratio of 3:1. The reaction mixture was stirred at room temperature for 1 h. Solid K₂CO₃ was slowly added to the reaction mixture to adjust the pH to 8–10. Water was added and the aqueous phase was extracted with DCM. The combined organic phases were purified by column chromatography to provide 12 as a yellow solid, 87 mg, in 84% yield. 1 H NMR (600 MHz, DMSO) δ 8.19 (d, J =2.1 Hz, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.86 (dd, J = 21.3, 8.4 Hz, 2H), 7.66(dd, J = 8.3, 2.1 Hz, 1H), 7.61 (dd, J = 8.4, 2.1 Hz, 1H), 7.53 (dd, J = 8.7, 2.1 Hz, 1H), 7.35 (d, J = 2.1 Hz, 1H), 7.14 (d, J = 8.7 Hz, 1H), 3.75 (s,3H), 3.51 (dd, J = 6.8, 4.4 Hz, 2H), 3.24 (dd, J = 46.9, 12.6 Hz, 2H), 2.79(ddd, J = 23.0, 13.8, 10.8 Hz, 2H), 2.20 (d, J= 13.7 Hz, 1H), 1.77 – 1.64 (m, 2H), 1.39 – 1.19 (m, 3H).

[0228] Synthesis of 4-chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-(piperidin-4-ylmethyl)phenyl)sulfonamide)benzoic acid (C3-332)

[0229] Intermediate 12 was dissolved in 5 mL of 1,4-dioxane and stirred at 40°C. 5 mL of 1N NaOH solution was added to the reaction mixture and stirred for 2 h. The dioxane was removed under vacuum, and the product on the container walls was dispersed into the solution using sonication. The reaction mixture was acidified to pH 6–7 with 1N HCl solution. The product was precipitated from water and stirred at room temperature for approximately 30 min. The precipitate was collected by filtration and washed with a small amount of water. The filter cake was dried in an oven to provide C3-332 as a yellow solid, 61.5 mg, 79% yield. 1 H NMR (400 MHz, DMSO) δ 8.26 (d, J = 2.0 Hz, 1H), 8.00 (d, J = 2.1 Hz, 1H), 7.85 (d, J = 8.3 Hz, 1H), 7.65 (ddd, J = 8.7, 4.0,2.1 Hz, 2H), 7.61 (d, J = 8.3 Hz, 1H), 7.41 (dd, J = 8.3, 2.1 Hz, 1H), 7.27(d, J = 2.1 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 3.14 (s, 4H), 2.71 (d, J =12.4 Hz, 2H), 1.85 (d, J = 66.9 Hz, 4H), 1.21 (d, J = 26.5 Hz, 2H).

[0230] Option 5. Synthesis of C3-341-342 and 345

[0231] Synthesis of methyl 2-((N-(3-((tert-butoxycarbonyl)amino)propyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)-4-chlorobenzoate (13)

[0232] Compound 13 was prepared from intermediate 6d (1.0 g, 1.83 mmol, 1 equivalent) and tert-butyl (3-bromopropyl)carbamate (1.7 g, 7.3 mmol, 4 equivalent) using the same method as for compound 11 in scheme 4. Yellow solid, 785 mg, 61% yield. 1 H NMR (400 MHz, DMSO) δ 8.26 (d, J = 2.1 Hz, 1H), 8.02 (d, J = 2.1 Hz, 1H), 7.87 (t, J = 8.7 Hz, 2H), 7.67 (dd, J = 8.3, 2.1 Hz, 1H), 7.60(td, J = 8.9, 2.1 Hz, 2H), 7.29 (d, J = 2.1 Hz, 1H), 7.16 (d, J = 8.6 Hz, 1H), 6.79 (t, J = 5.7 Hz, 1H), 3.75 (s, 3H), 3.70 – 3.49 (m, 2H), 2.93 (q, J = 6.2 Hz, 2H), 1.58 (d, J = 53.3 Hz, 2H), 1.35 (s, 9H).

[0233] Synthesis of methyl 2-((N-(3-aminopropyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamide)-4-chlorobenzoate (14)

[0234] Compound 14 was prepared from intermediate 13 (770 mg, 1.09 mmol, 1 equivalent) using the same method as for compound 12 in scheme 4. Yellow solid, 574 mg, 87% yield. 1 H NMR (400 MHz, DMSO) δ 8.26 (d, J = 2.1 Hz, 1H), 8.03 (d,J = 2.0 Hz, 1H), 7.89 (d, J = 8.3 Hz, 1H), 7.86 (d, J = 8.5 Hz, 1H), 7.68 (dd, J = 8.3, 2.1 Hz, 1H), 7.61 (td, J = 8.9, 2.1 Hz, 2H), 7.29 (d, J = 2.0 Hz, 1H), 7.17 (d, J = 8.6 Hz, 1H), 5.77 (s, 1H), 3.76(s, 3H), 3.57 (s, 2H), 2.61 (t, J = 6.8 Hz, 2H), 1.56 (d, J = 42.3 Hz, 2H).

[0235] Synthesis of methyl 2-((N-(3-acetamidopropyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-chlorobenzoate (15)

[0236] DIPEA (165 μL, 0.94 mmol, 3 equivalents) was added to intermediate 14 (190 mg, 0.31 mmol, 1 equivalent) dissolved in DCM. Acetic anhydride (45 μL, 0.47 mmol, 1.5 equivalents) was added to the reaction mixture and stirred at room temperature for 2 h. Water was added and the aqueous phase was extracted with DCM. The combined organic phases were purified by column chromatography to provide 15 as a yellow solid, 122 mg, in 60% yield. 1 H NMR (400 MHz, DMSO) δ 8.25 (d, J = 2.1 Hz, 1H), 8.03(d, J = 2.1 Hz, 1H), 7.88 (d, J = 8.3 Hz, 1H), 7.86 (d, J = 8.4 Hz, 1H), 7.79(t, J = 5.6 Hz, 1H), 7.68 (dd, J = 8.4, 2.1 Hz, 1H), 7.61 (ddd, J= 8.9, 6.9,2.1 Hz, 2H), 7.31 (d, J = 2.1 Hz, 1H), 7.17 (d, J = 8.6 Hz, 1H), 3.75 (s,3H), 3.70 – 3.50 (m, 2H), 3.03 (q, J = 6.7 Hz, 2H), 1.75 (s, 3H), 1.67 – 1.49 (m, 2H).

[0237] General methods for synthesizing C3-341, 342 and 345 C3-341-342 and 345 were prepared from the corresponding intermediates using the same method as that used for C3-332 in Scheme 4.

[0238] 2-((N-(3-((tert-butoxycarbonyl)amino)propyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-chlorobenzoic acid (C3-341)

[0239] Yellow solid, 152 mg, 86% yield. 1 H NMR (600 MHz, DMSO) δ 13.22 – 13.05 (m,1H), 8.28 (d, J = 2.1 Hz, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.86 (dd, J = 17.5, 8.3 Hz, 2H), 7.67 (dd, J = 8.3, 2.1 Hz, 1H), 7.62 – 7.56 (m, 2H), 7.28 – 7.23(m, 1H), 7.13 (d, J = 8.6 Hz, 1H), 6.78 (t, J = 5.8 Hz, 1H), 3.64 – 3.53 (m,2H), 2.90 (d, J = 6.4 Hz, 2H), 1.56 (d, J = 79.1 Hz, 2H), 1.34 (s, 9H). 13C NMR(151 MHz, DMSO) δ 166.46, 156.00, 144.77, 143.09, 138.75, 137.22, 136.81,136.10, 135.99, 134.48, 133.38, 133.08, 132.98, 132.38, 132.27, 130.75, 130.05, 129.36, 125.02, 77.99, 49.59, 37.72, 28.66, 22.54.

[0240] Synthesis of 2-((N-(3-aminopropyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-chlorobenzoic acid (C3-342)

[0241] Yellow solid, 50 mg, 27% yield. 1 H NMR (600 MHz, DMSO) δ 8.37 (d, J = 2.1 Hz, 1H), 8.02 (d, J = 2.1 Hz, 1H), 7.85 (d, J = 8.3 Hz, 1H), 7.74 (dd, J = 8.6, 2.1 Hz, 1H), 7.67 (dd, J = 8.3, 2.1 Hz, 1H), 7.51 (d, J = 8.3 Hz, 1H), 7.35 (dd, J = 8.3, 2.1 Hz, 1H), 7.13 (d, J = 8.6 Hz, 1H), 7.02 (d, J = 2.1 Hz,1H), 3.62 (s, 2H), 2.79 (t, J = 6.6 Hz, 2H), 1.68 (s, 2H).

[0242] Synthesis of 2-((N-(3-acetamidopropyl)-4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)-4-chlorobenzoic acid (C3-345)

[0243] Yellow solid, 101 mg, 88% yield. 1H NMR (600 MHz, DMSO) δ 8.28 (d, J = 2.1 Hz, 1H), 8.02 (d, J = 2.1 Hz, 1H), 7.87 (d, J = 8.3 Hz, 1H), 7.83 (d, J = 8.4 Hz, 1H), 7.79 (t, J = 5.6 Hz, 1H), 7.67 (dd, J = 8.3, 2.1 Hz, 1H), 7.62 (dd, J =8.7, 2.1 Hz, 1H), 7.58 (dd, J = 8.4, 2.1 Hz, 1H), 7.26 (d, J = 2.1 Hz, 1H), 7.14 (d, J = 8.7 Hz, 1H), 3.58 (d, J = 9.7 Hz, 2H), 3.00 (q, J = 6.6 Hz, 2H), 1.74 (s, 3H), 1.56 (d, J = 47.5 Hz, 2H). 13 C NMR (151 MHz, DMSO) δ 169.48,166.54, 144.75, 143.09, 138.63, 137.23, 136.68, 135.99, 134.47, 133.37,133.02, 132.97, 132.43, 130.73, 130.64, 130.05, 129.35, 125.09, 49.53, 36.46,28.71, 22.99.

[0244] C3-347 Synthesis Route

[0245] 4-Chloro-2-((4-chloro-3-nitrophenyl)sulfonamido)methyl benzoate (4d)

[0246] Pyridine (9.76 mL, 121.20 mmol, 7.50 equivalents) was added to a solution of methyl 2-amino-4-chlorobenzoate (3.00 g, 16.20 mmol, 1.00 equivalents) in DCM (25 mL) at 0°C. A solution of 4-chloro-3-nitrobenzenesulfonyl chloride (5.00 g, 19.40 mmol, 1.20 equivalents) in DCM (25 mL) was added dropwise to the mixture at 0°C. The mixture was heated to room temperature and stirred overnight at 25°C. After the reaction was complete, water was added to quench the reaction, and the resulting solution was extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by column chromatography (hexane / EtOAc = 10 / 1) to give the title compound. Pale yellow solid (2.47 g, 37.76%). 1 H NMR (400 MHz, DMSO) δ 10.69 (s, 1H), 8.46 (d, J = 2.1 Hz, 1H), 8.02 (dd, J = 8.5, 2.2 Hz, 2H), 7.97 (d, J = 8.5 Hz, 1H), 7.42 (d, J = 2.1Hz, 1H), 3.75 (s, 4H).

[0247] Methyl 4-chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)benzoate (6d)

[0248] Add NaOAc (2.50 g, 30.50 mmol, 15.00 equivalents) to a solution of methyl 4-chloro-2-((4-chloro-3-nitrophenyl)sulfonamido)benzoate (2.47 g, 6.10 mmol, 3.00 equivalents) in EtOH (20 mL). Add 3,4-dichlorothiophenol (1.82 g, 10.17 mmol, 5.00 equivalents) to the mixture. [The solution was prepared at 80 °C.] The mixture was stirred at C for 6 hours. After the reaction was complete, water was added to quench the reaction, and the resulting solution was extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by column chromatography (hexane / EtOAc = 10 / 1) to obtain the title compound. Yellow solid (2.89 g, 86.44%). 1H NMR (400 MHz, DMSO) δ10.66 (s, 1H), 8.52 (d, J = 2.1 Hz, 1H), 8.01 (d, J = 2.1 Hz, 1H), 7.90 –7.82 (m, 3H), 7.65 (dd, J = 8.3, 2.1 Hz, 1H), 7.43 (d, J = 2.1 Hz, 1H), 7.36(d, J = 8.5 Hz, 1H), 7.15 (d, J = 8.7 Hz, 1H), 3.76 (s, 3H).

[0249] 4-Chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-(prop-2-yn-1-yl)phenyl)sulfonamido)methyl benzoate (16)

[0250] Potassium carbonate (150.00 mg, 1.10 mmol, 2.00 equivalent) was added to a solution of methyl 4-chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)benzoate (300.00 mg, 0.55 mmol, 1.00 equivalent) in DMF (1.80 mL). 3-Bromoprop-1-yne (70.00 mL, 0.82 mmol, 1.50 equivalent) was added to the mixture. The mixture was stirred overnight at 80°C. Water was added to quench the reaction, and the resulting solution was extracted with EtOAc (3 × 10 mL). The combined organic layers were washed with brine and dried over anhydrous Na₂SO₄. The solution was then filtered through a short silica gel sieve, washed with EtOAc, and concentrated under vacuum. The residue was redissolved in DCM (2 mL), followed by the addition of hexane (20 mL) and Et₂O (1 mL), and stirred at room temperature for 10 minutes. A yellow precipitate was filtered off, and the resulting solid was washed with hexane / Et₂O (v / v = 3 / 1) to obtain the title compound. Light brown solid (268 mg, 83.53%). 1 H NMR (400 MHz, CDCl3) δ8.65 (d, J = 2.1 Hz, 1H), 7.88 (d, J = 8.4 Hz, 1H), 7.71 (d, J= 2.1 Hz, 1H),7.64 – 7.60 (m, 2H), 7.47 – 7.42 (m, 2H), 7.18 (d, J = 2.1 Hz, 1H), 6.95 (d, J = 8.7 Hz, 1H), 3.83 (s, 3H), 2.35 (d, J = 5.0 Hz, 1H).

[0251] 4-Chloro-2-((4-((3,4-dichlorophenyl)thio)-3-nitro-N-(prop-2-yn-1-yl)phenyl)sulfonamido)benzoic acid (C3-347)

[0252] A solution of methyl 4-chloro-2-((4-(((3,4-dichlorophenyl)thio)-3-nitrophenyl)sulfonamido)benzoate (268 mg, 0.46 mmol, 1.00 equivalent) in 1,4-dioxane (4 mL) was added to 1 N NaOH (4 mL), and the mixture was stirred at 40°C for 3 h. After the reaction was complete, the mixture was acidified to pH 3 with 1 N HCl. The resulting suspension was extracted with DCM (3 × 10 mL). The combined organic layers were washed with brine, dried over anhydrous Na₂SO₄, filtered, and evaporated under vacuum. The residue was purified by semi-preparative HPLC [15–100% (ACN) / (0.1% HCOOH, in H₂O)] to obtain the title compound. Yellow solid (18.9 mg, 6.45%). 1 H NMR (400 MHz, CDCl3) δ 8.68 (d, J = 2.0 Hz, 1H), 7.99 (d, J = 8.5 Hz, 1H), 7.69 (d, J = 2.1 Hz, 1H), 7.59 (dd, J = 8.5, 2.0Hz, 2H), 7.52 (dd, J = 8.4, 2.1 Hz, 1H), 7.42 (dd, J = 8.3, 2.1 Hz, 1H), 7.26(s, 1H), 6.95 (d, J = 8.6 Hz, 1H), 4.94 (s, 1H), 4.23 (s, 1H), 2.37 (t, J =2.4 Hz, 1H).13 C NMR (151 MHz, CDCl3) δ 169.05, 144.30, 144.28, 139.36, 138.70, 137.15, 137.06, 135.83, 134.81, 134.61, 133.48, 132.46, 132.37, 131.66, 130.10, 129.56, 128.49, 125.51, 77.60, 75.03, 41.82. HRMS (ESI-TOF) m / z: for [C 22 H 12 Cl3N2O6S2] - ([M - H] - Calculated value: 568.9208, measured value: 568.9214. HPLC purity: 97.30%.

[0253] 4.2 Biology 4.2.1. Determination of the minimum inhibitory concentration (MIC) The antimicrobial activity of the compound was determined by broth microdilution according to the Clinical & Laboratory Standards Institute (CLSI) guidelines (2020). The test medium was cationic-regulated Mueller-Hinton broth (MHB) (unless otherwise specified) or brain-heart infusion (BHI) broth (as indicated in the text). For the chemical being tested, sequential 2-fold dilutions of the test compound were performed, starting at 256 µg / mL and decreasing to 0.25 μg / mL, and the bacterial inoculum was adjusted to approximately 5 × 10⁻⁶. 5 CFU / mL. Results were obtained after incubation at 37°C for 20 hours. MIC was defined as the lowest concentration of antibiotic that produces no visible growth. Experiments were performed in duplicate.

[0254] Antimicrobial activity of C3-332, C3-341, C3-342, C3-345 and C3-347 We examined the antibacterial spectrum of representative compounds. The bacterial group we used consisted of three Gram-positive pathogens from the latest version of the WHO Priority Pathogens List for Guiding the Development of New Antibiotics: Enterococcus faecalis, Staphylococcus aureus, Streptococcus pneumoniae, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter cloacae, and Escherichia coli. Overall, the compounds demonstrated broad-spectrum antimicrobial activity against all tested Gram-positive bacteria, as shown in Table 5.

[0255] Table 5. Antibacterial activity (MIC mg / ml) of compound C3 against nine representative bacterial strains on the WHO priority pathogen list used to guide the development of new antibiotics.

[0256] 4.2.2. Determination of inhibitory activity against β'CH-σ interaction The previously established protocol was used for inhibitor assays (Tsang et al., 2019). Protein overproduction and purification were performed as described previously (Tsang et al., 2019). C-SmBiT-CH (40 μL, 2.5 μM, in PBS) was added to a 96-well plate and then mixed with 20 μL of the desired concentration of the compound. The mixture was incubated at 37°C for 10 min. Then C-LgBiT-σ... A (40 μL, 2.5 μM, in PBS) was added to each well, followed by incubation at 37°C for 10 minutes. After the final incubation step, an equal volume of Promega Nano-Glo® luciferase assay substrate (Promega, Madison, Wisconsin, United States) was added to the reaction mixture. The emitted light was measured using a Victor X3 Multilabel plate reader (Waltham, Massachusetts, United States). For consistent results, experiments were repeated using a triplicate technique.

[0257] 4.2.3. Epifluorescence microscopy Bacillus subtilis strain BS1048 was grown on LB agar plates containing 5 mg / mL chloramphenicol and 0.5% (w / v) xylose. Single colonies were incubated at 37°C in LB medium containing 5 mg / mL chloramphenicol and 0.5% (w / v) xylose until OD (digestive activity) was reached. 600 ~0.6. Chloramphenicol, rifampin, or the compound was then added to the culture and incubated for an additional 15 minutes. 2.5 μL of cell culture was placed onto a 1.2% freshly prepared agarose plate, covered with a coverslip, and then imaged. A Nikon Eclipse Ti2-E live-cell fluorescence imaging system equipped with a 63 × 1.4 oil immersion lens and LED illumination was used to capture fluorescence images. Fluorescence images were processed using Nikon NIS-Elements and MetaMorph software.

[0258] 4.2.4. Molecular docking The compound was derived from the structure of Escherichia coli RNAP (PDB: 4LJZ) to obtain β Molecular docking in CH was performed using AutoDock Vina. AutoDock Tools-1.5.7 was used to prepare ligands and macromolecules to generate PDBQT files. The binding pocket was defined by a center at (8.242, -53.403, -46.042) and a size of (13, 13, 13). Finally, the docking conformation was visualized using Pymol Academic Edition (Schrödinger, LLC).

[0259] 4.2.5. Cytotoxicity Assay Lung cancer A549 and hepatocellular carcinoma HaCaT cell lines were used to evaluate the cytotoxicity of antimicrobial compounds, and assays were performed as previously described. On day 0, cells were sputtered at a rate of 2.5 × 10⁻⁶. 5 Cells were seeded per well in 96-well plates. After overnight incubation at 37°C for 24 hours, the test compound and positive control cisplatin were added at serially diluted two-fold concentrations ranging from 1.562 mg / mL to 50 μg / mL. The plates were then incubated at 37°C, and cell viability was determined by CCK-8 assay at 48 and 72 hours after compound addition. Results are expressed as 50% cytotoxic concentration (CBC). 50 The values ​​and their corresponding standard deviations refer to the concentrations of compounds that reduce cell viability by 50% compared to the untreated control.

Claims

1. A compound of formula 1: 1 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 3; n is an integer selected from 1 to 3; p is an integer selected from 1 to 4; X is -S-, -O-, -C(R)2- or -N(R-); Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl or -(CR 6 2) q Y, where q is an integer selected from 0 to 6; R 6 Each occurrence of R is independently selected from hydrogen, alkyl, and cycloalkyl; or two occurrences of R. 6 Together with the covalently bonded carbon atoms, they form 3-6 membered cycloalkyl groups; and Y is an alkynyl, optionally carbonyl-substituted cycloalkyl, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; R 3 It is hydrogen or alkyl; R 4 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; and R 5 Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R. The condition is that the compound of formula 1 is not: 。 2. The compound according to claim 1, wherein X is -S- or -O-.

3. The compound according to claim 1, wherein R 5 Each time it appears, it is independently selected from alkyl, haloalkyl, perhaloalkyl, perhaloalkoxy, and halides.

4. The compound according to claim 1, wherein R 4 Each time it appears, it is independently selected from hydrogen, fully halogenated alkyl, nitrile, nitro, -C(O)R, -C(O)OR, -C(O)N(R)2, -S(O)2R and -S(O)2N(R)2.

5. The compound according to claim 1, wherein R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2 and -C(O)N(R)2.

6. The compound according to claim 1, wherein the compound has formula 2: 2 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 3; n is an integer selected from 1 to 2; p is an integer selected from 1 to 2; X is -S- or -O-; Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is alkyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl or -(CR 6 2) q Y, where q is an integer selected from 0 to 6; R 6 Each occurrence of R is independently selected from hydrogen, alkyl, and cycloalkyl; or two occurrences of R. 6 Together with the covalently bonded carbon atoms, they form 3-6 membered cycloalkyl groups; and Y is an alkynyl, optionally carbonyl-substituted cycloalkyl, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2 and -C(O)N(R)2; R 3 It is hydrogen or alkyl; R 4 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; R 5 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 and -N(R)S(O)2R; R 7 It is a fully haloalkyl, nitrile, nitro, -C(O)R, -C(O)OR, -C(O)N(R)2, -S(O)2R or -S(O)2N(R)2; and R 8 Each time it appears, it is independently selected from alkyl, haloalkyl, perhaloalkyl, perhaloalkoxy, and halides.

7. The compound according to claim 6, wherein R 1 It is a cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, or -(CH2) group. q Y, where q is an integer selected from 0-6; and Y is an alkynyl group, a cycloalkyl group optionally substituted with a carbonyl moiety, a heteroaryl group, a heterocycloalkyl group, a 3,3-difluoropyrrolidone-1-yl group, N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R.

8. The compound according to claim 1, wherein the compound has formula 3: 3 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 3; X is -S- or -O-; Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is a cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, or -(CH2) group. q Y, where q is an integer selected from 0-6, and Y is an alkynyl group, a cycloalkyl group optionally substituted with a carbonyl moiety, a heteroaryl group, a heterocycloalkyl group, or a 3,3-difluoropyrrolidone-1-yl group. N -phthalimide, arginine, -CN, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2 and -C(O)N(R)2; R 3 It is hydrogen or a C1-C3 alkyl group; R 4 It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; and R 5 It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R.

9. The compound according to claim 8, wherein R 2 Each time it appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, halide, nitrile, nitro, -OR and -SR.

10. The compound according to claim 8, wherein m is 1 or 2; and R 2 Each time it appears, it is independently selected from hydrogen, alkyl, fully haloalkyl, halide and -O-(C1-C6 alkyl).

11. The compound according to claim 8, wherein R 1 It is an aralkyl group or -(CH2) q Y, where q is an integer selected from 0-5, and Y is an alkynyl group, a cycloalkyl group optionally substituted with a carbonyl moiety, a heteroaryl group, a heterocycloalkyl group, or a 3,3-difluoropyrrolidone-1-yl group. N -phthalimide, arginine, -CN, -OR, -C(O)R, -C(O)OR, -N(R)C(O)R, -N(R)C(O)OR, -S(O)2R, -S(O)2N(R)2 or -N(R)2.

12. The compound according to claim 1, wherein the compound has formula 4: 4 Or its pharmaceutically acceptable salt, wherein m is an integer selected from 1 to 2; Each time R appears, it is independently selected from hydrogen, alkyl, haloalkyl, perhaloalkyl, ether, alkenyl, ynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, and aralkyl; or R appearing twice together with their covalently bonded atoms form 3-6 membered cycloalkyl or heterocycloalkyl groups. R 1 It is an aralkyl group or -(CH2) q Y, where q is an integer selected from 0-5, and Y is an alkynyl, -OR, -CN, cycloalkyl optionally substituted with a carbonyl group, heteroaryl, heterocycloalkyl, 3,3-difluoropyrrolidone-1-yl, -C(O)OR, N -phthalimide, arginine, -CN, -OR, -C(O)R, -N(R)C(O)OR, -S(O)2R, -S(O)2N(R)2 or -N(R)2; R 2 Each time it appears, it is independently selected from hydrogen, alkyl, fully haloalkyl, halide or -O- (C1-C6 alkyl); R 3 It is hydrogen or C1-C3 alkyl; R 4 It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)-N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R; and R 5 It is hydrogen, alkyl, haloalkyl, perhaloalkyl, alkenyl, alkynyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, aralkyl, perhaloalkoxy, halide, nitrile, nitro, azide, -OR, -SR, -N(R)2, -C(O)R, -C(O)OR, -OC(O)R, -N(R)C(O)R, -C(O)N(R)2, -N(R)C(O)OR, -OC(O)N(R)-, -OC(O)OR, -N(R)C(O)N(R)2, -S(O)2R, -S(O)2N(R)2 or -N(R)S(O)2R.

13. The compound according to claim 12, wherein R 3 R 4 and R 5 Each is hydrogen.

14. The compound according to claim 12, wherein R 1 Selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and .

15. The compound according to claim 12, wherein R 2 Each time it appears, it is independently selected from hydrogen, methyl, fluoride, bromide, chloride, -CF3 or -OCH3.

16. The compound according to claim 12, wherein R 1 Selected from: , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , and ; R 2 Each time it appears, it is independently selected from hydrogen, methyl, fluoride, bromide, chloride, -CF3, or -OCH3; and R 3 It is hydrogen.

17. The compound according to claim 1, wherein the compound has formula 5: 5 Or a pharmaceutically acceptable salt thereof, wherein R 1 Selected from: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 , , , , , , , , , , , , , and .

18. The compound according to claim 17, wherein R 1 Selected from: , , , , , , , , and .

19. The compound according to claim 1, wherein the compound is selected from: , , , , And its pharmaceutically acceptable salts.

20. A pharmaceutical composition comprising the compound according to claim 1 and at least one pharmaceutically acceptable carrier or pharmaceutically acceptable excipient.

21. A method of treating a bacterial infection in a subject in need, the method comprising administering to the subject a therapeutically effective amount of the compound according to claim 1.

22. The method of claim 21, wherein the bacterial infection is caused by Gram-positive bacteria.

23. The method of claim 21, wherein the bacterial infection is caused by Enterococcus faecalis (… Enterococcus faecalis Staphylococcus epidermidis ( Staphylococcus epidermidis ), saprophytic Staphylococcus ( Staphylococcus saprophyticus ), Streptococcus pyogenes ( Streptococcus pyogenes ), agalactococcus ( Streptococcus agalactiae ), Enterococcus faecalis ( Enterococcus faecalis Staphylococcus aureus ( Staphylococcus aureus Streptococcus pneumoniae () Streptococcus pneumoniae It is caused by infection with methicillin-resistant Staphylococcus aureus or vancomycin-resistant Staphylococcus aureus.

24. The method of claim 21, wherein the bacterial infection is caused by antibiotic-resistant Staphylococcus aureus infection or antibiotic-resistant Streptococcus pneumoniae infection.

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