2-(2-(2-benzoylamino)acetamido)benzoic acid derivatives, processes for their preparation and uses thereof

Abstract

The present application discloses 2-(2-(2-benzoylamino)acetamido)benzoic acid derivatives or its pharmaceutically acceptable salt as a bacterial type I signal peptidase inhibitor, which has a broad spectrum of biological activity. In various embodiments, the present application compound exerts its effect by inhibiting bacterial type I signal peptidase (SPase), which plays a key role in the survival and virulence of bacteria. The present application also provides a preparation method of the compound and its pharmaceutically acceptable salt, a pharmaceutically acceptable carrier or excipient.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and particularly relates to a 2-(2-(2-benzamido)acetamide)benzoic acid derivative, a preparation method and use thereof, and the compound can be used as a type I signal peptidase inhibitor of bacteria. BACKGROUND

[0002] The discovery and application of antibiotics is one of the milestones of modern medicine, which has made outstanding contributions to saving human lives and improving human health. However, with the overuse or misuse of antibiotics in the medical and agricultural fields, the problem of bacterial drug resistance has become increasingly serious. Therefore, the demand for new antibiotics is imminent.

[0003] Danish microbiologist Hans Christian Gram discovered in the late 19th century that bacteria can be divided into gram-positive bacteria and gram-negative bacteria. The cell wall of gram-positive bacteria is composed of a thick and dense peptidoglycan and phosphomannan, while the cell wall of gram-negative bacteria is a multi-layer structure, from inside to outside, in turn: a thin peptidoglycan layer, a lipoprotein layer / periplasmic layer, a phospholipid layer and a lipopolysaccharide layer. This special structure leads to the fact that many antibiotics (such as vancomycin) can reach their targets through the outer peptidoglycan surface of gram-positive bacteria, but cannot pass through the outer membrane of gram-negative bacteria. For nearly half a century, there has still been no antibiotic approved globally that can effectively target gram-negative bacteria. Coincidentally, most pathogenic bacteria that produce multiple drug resistance, so-called superbugs, also belong to gram-negative bacteria. It is worrying that not only the number of weapons is small, but also many of the weapons we have are becoming increasingly ineffective. Once these drug-resistant bacteria infect the human body, the consequences will be very dangerous and even fatal without any effective drugs available.

[0004] The imminent antibiotic crisis has prompted people to develop new strategies to fight infections. All bacteria need to export proteins through the cytoplasmic membrane. Most proteins are achieved through the general secretory pathway (Sec), and the last step is to release the mature protein through the action of type I signal peptidase. Therefore, type I signal peptidase is essential, and it is highly conserved in bacteria, making type I signal peptidase inhibitors have the potential for broad-spectrum antibacterial activity. Due to the very important characteristics of type I signal peptidase, it has become a target for developing new antibiotics. SUMMARY

[0005] The present application aims to provide a 2-(2-(2-benzamido)acetamide)benzoic acid derivative or a pharmaceutically acceptable salt thereof as shown in general formula I:

[0006]

[0007] wherein,

[0008] R 1 selected from hydrogen, nitro,

[0009] R 2 selected from hydrogen, Boc protected or unprotected C 4-6 amino methyl;

[0010] R 3 selected from hydrogen,

[0011] R 4 selected from hydrogen, methyl, ethyl, propyl, isopropyl or isobutyl;

[0012] R 5 selected from carboxyl,

[0013] R 6 selected from hydrogen or C 4-6 containing C

[0014] Ar is an aromatic compound, including benzene rings and other aromatic heterocycles;

[0015] n is a natural number less than 20.

[0016] In some preferred embodiments, the pharmaceutically acceptable salts include, but are not limited to, acid addition salts of the compounds of Formula I with hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, maleic acid, benzenesulfonic acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, fumaric acid, salicylic acid, phenylacetic acid or mandelic acid; also acid salts of the compounds of Formula I with inorganic bases or organic salts made from basic amines.

[0017] The compounds of Formula I according to the present application are preferably the following compounds:

[0018]

[0019]

[0020] The compounds of Formula I according to the present application can also exist in the form of their salts, which are converted in vivo into the compounds of Formula I. For example, within the scope of the present application, the compounds according to the present application are converted into pharmaceutically acceptable salt forms according to processes known in the art and used as salts.

[0021] It is another object of the present application to provide a process for the preparation of the compounds of Formula I, comprising the following steps:

[0022] It is another object of the present application to provide a process for the preparation of the compounds of Formula I, comprising the following steps:

[0023] Step one: compound II reacts with methyl 2-aminobenzoate or methyl 3- aminoisonicotinate to obtain compound III;

[0024] Step two: compound III reacts in the presence of a condensing agent to obtain compound IV;

[0025] Step three: compound IV reacts with an intermediate in the presence of a condensing agent to obtain compound V;

[0026] Step four: compound V is reduced and then reacts with acyl chlorides of different carbon chain lengths to obtain compound VI;

[0027] Step five: compound VI reacts under acidic or basic conditions to obtain compound VII;

[0028] Step six: compound VII reacts in the presence of a condensing agent to obtain the target compound.

[0029] In some preferred embodiments, the method for preparing the intermediate comprises the following steps:

[0030]

[0031] Step one: compound VIII reacts with NBS to obtain compound IX;

[0032] Step two: compound IX is subjected to Gabriel reaction and tert-butyloxycarbonyl protection of the amino group to obtain compound X;

[0033] Step three: compound X reacts under acidic or basic conditions to obtain the intermediate.

[0034] wherein, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 are as defined in general formula I.

[0035] The compounds of general formula I of the present application can be prepared by the above-mentioned or similar methods, and corresponding starting materials can be selected according to the differences of substituents and the positions of substituents. Those skilled in the art should recognize that the above-mentioned routes are helpful for understanding the present application but do not limit the content of the present application, and the variables are defined as mentioned in general formula I, unless otherwise specified.

[0036] Another object of the present application is to provide a pharmaceutical composition comprising a 2-(2-(2-benzamido)acetamido)benzoic acid derivative of general formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

[0037] The pharmaceutical compositions of the present application can be administered in a variety of known ways, e.g., orally, parenterally, by inhalation spray, or via an implanted reservoir. The pharmaceutical compositions of the present application can be administered alone or in combination with other drugs. Oral compositions can take a variety of forms, including but not limited to tablets, capsules, pills, and suspensions, gelcaps, and oral liquids. Pharmaceutically acceptable carriers are those conventional in the art, such as one or more inert, non-toxic solid or liquid fillers, diluents, excipients, and the like, which are not toxic to the host and do not interact deleteriously with the active compound or the patient. Common pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, binders, colorants, fillers, emulsifiers, and the like.

[0038] Injectable solutions can be prepared using appropriate dispersing or wetting agents and suspending agents, as known in the art. Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride, and the like.

[0039] Actual dosage levels of the active ingredients in the pharmaceutical compositions of this application can be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, compositions, and mode of administration, without being toxic to the patient. The selected dosage level will depend on a variety of factors including the activity of the particular compound of the present application or salt thereof employed, the route of administration, the time of administration, the rate of excretion of the particular compound being employed, the duration of the treatment, other drugs, compounds, and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.

[0040] It is another object of the present application to provide the use of a compound of Formula I or a pharmaceutically acceptable salt thereof in the manufacture of a medicament for the treatment of a bacterial infection. The 2-(2-(2-benzamido)acetamido)benzoic acid derivatives of the present application or a pharmaceutically acceptable salt thereof are capable of acting as inhibitors of bacterial type I signal peptidase.

[0041] The infection or bacterial infection refers to a disease or condition characterized by the invasion of the body tissues of an organism by a pathogenic agent (e.g., pathogenic bacteria), their multiplication, and the reaction of the host tissues to the infecting agent and its produced toxins. Infectious diseases, also known as transmissible diseases, are diseases caused by infectious agents. The bacterial infection can be caused by gram-positive or gram-negative bacteria.

[0042] The bacterial infection includes diseases caused by infection of Elizabethkingia meningitidis, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas acidovorans, Pseudomonas alcaligenes, Pseudomonas putida, Stenotrophomonas maltophilia, Burkholderia cepacia, Aeromonas hydrophila, Escherichia coli, Citrobacter freundii, Salmonella typhimurium, Salmonella typhi, Salmonella paratyphi, Salmonella enteritidis, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Enterobacter cloacae, Enterobacter aerogenes, Klebsiella pneumoniae, Klebsiella oxytoca, Serratia marcescens, Francisella tularensis, Morganella morganii, Proteus mirabilis, Proteus vulgaris, Providencia alcalifaciens, Providencia rettgeri, Providencia stuartii, Acinetobacter baumannii, Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Yersinia enterocolitica, Yersinia pestis, Yersinia pseudotuberculosis, Yersinia intermedia, Bordetella pertussis, Bordetella parapertussis, Bordetella bronchiseptica, Haemophilus influenzae, Haemophilus parainfluenzae, Haemophilus haemolyticus, Haemophilus parahaemolyticus, Haemophilus ducreyi, Pasteurella multocida, Pasteurella haemolytica, Branhamella catarrhalis, Helicobacter pylori, Campylobacter fetus, Campylobacter jejuni, Campylobacter coli, Borrelia burgdorferi, Vibrio cholerae, Vibrio parahaemolyticus, Legionella pneumophila, Listeria monocytogenes, Neisseria gonorrhoeae, Neisseria meningitidis, Chlamydia, Moraxella, Gardnerella vaginalis, Bacteroides fragilis, Bacteroides distasonis, Bacteroides vulgatus, Bacteroides ovalus, Bacteroides thetaiotaomicron, Bacteroides uniformis, Bacteroides eggerthii, Bacteroides splanchnicus, Clostridium difficile, Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium leprae, Corynebacterium diphtheriae, Corynebacterium ulcerans, Streptococcus pneumoniae, Streptococcus agalactiae, Streptococcus pyogenes, Enterococcus faecalis, Enterococcus faecium, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Staphylococcus intermedius, Streptococcus suis, Staphylococcus hyicus subsp. hyicus, Staphylococcus haemolyticus, or Staphylococcus hominis.

[0043] Beneficial effects:

[0044] (1) The compound of general formula I and its pharmaceutically acceptable salt prepared by the present application has excellent bacterial type I signal peptide enzyme inhibition effect. The compound can inhibit bacterial type I signal peptide enzyme, block the Sec and Tat secretion system, and cause the bacteria to be unable to release mature secretory proteins, thereby playing an antibacterial role. Therefore, the above-mentioned compound can be used for preparing a drug for preventing, treating or improving bacterial infection. For example, the compound can be used for preparing a drug for treating diseases caused by infection of Staphylococcus aureus, Enterococcus faecalis, Streptococcus pyogenes, Escherichia coli and other bacteria.

[0045] (2) The synthesis route of the compound of the present application is simple, easy to implement, and easy to realize industrial production. DETAILED DESCRIPTION

[0046] The technical solutions of the present application will be described in detail below with specific examples, but the protection scope of the present application is not limited to the examples.

[0047] In the following examples, "room temperature" means about 20-30°C. The ratio of the mixed solvents means the volume mixing ratio, and % means wt% unless otherwise specified. M means molar concentration.

[0048] In the following examples, "room temperature" means about 20-30°C. The ratio of the mixed solvents means the volume mixing ratio, and % means wt% unless otherwise specified. M means molar concentration.

[0049] In the silica gel column chromatography, basic silica gel means silica gel combined with aminopropylsilane. In the high performance liquid chromatography (HPLC), C18 means silica gel combined with octadecyl. The ratio of the elution solvents means the volume mixing ratio, and the elution solvents are petroleum ether, dichloromethane, ethyl acetate, and methanol unless otherwise specified.

[0050] In the following examples, the following abbreviations are used: tetrahydrofuran THF, dichloromethane DCM, dimethyl sulfoxide DMSO, N,N-diisopropyl ethylamine DIEA, potassium acetate KOAc, triethylamine Et3N.

[0051] MS (mass spectrum) was measured using LC / MS (liquid chromatograph mass spectrometer) using an ESI (electrospray ionization) method. In the case of a salt, a molecular ion peak or a fragment ion peak in a free form is usually observed. 1 H-NMR (proton nuclear magnetic resonance spectrum). The peaks of active hydrogen (e.g., hydroxyl group, amino group, etc.) are not described using software such as ACD / SpecManager.

[0052] MS (mass spectrum) was measured using LC / MS (liquid chromatograph mass spectrometer) using an ESI (electrospray ionization) method. In the case of a salt, a molecular ion peak or a fragment ion peak in a free form is usually observed.

[0053] Preparation of 3-(N-Boc-aminomethyl)benzoic acid in Example 1

[0054]

[0055] Methyl 3-methylbenzoate (1.0 g), N-bromosuccinimide (1.2 g), azobisisobutyronitrile (0.1 g), and 20 mL of carbon tetrachloride were refluxed for 12 hours. After suction filtration, the filtrate was dried under reduced pressure, and purified by silica gel column chromatography to obtain methyl 3-bromomethylbenzoate (1.2 g).

[0056] Methyl 3-bromomethylbenzoate (1.0 g), phthalimide (0.7 g), potassium carbonate (1.20 g) and 20 mL of dimethylformamide were reacted at room temperature for about 24 hours. After filtration with water, white solid was obtained. The white solid (0.6 g), hydrazine hydrate (0.4 g), p-toluenesulfonic acid monohydrate (0.03 g) and 20 mL of tetrahydrofuran were reacted at reflux for 4 hours. After removing insoluble matter by filtration, the filtrate was dried by evaporation to obtain methyl 3-aminomethylbenzoate (0.3 g).

[0057] Methyl 3-aminomethylbenzoate (0.3 g), di-tert-butyl dicarbonate (0.44 g), 5 mL of saturated sodium bicarbonate solution and 5 mL of dichloromethane were reacted at room temperature for about 24 hours. After adding 30 mL of water, 3 times of extraction with 30 mL of dichloromethane was performed, and the combined organic layer was dried under reduced pressure and purified by silica gel column chromatography to obtain methyl 3-(N-Boc-aminomethyl)benzoate (0.3 g).

[0058] Methyl 3-(N-Boc-aminomethyl)benzoate (0.3 g), lithium hydroxide (0.2 g), 4 mL of water and 8 mL of tetrahydrofuran were reacted at room temperature for about half an hour. After acidification with acetic acid by adding water, white solid was precipitated, and the target compound (0.2 g) was obtained by filtration as a white solid with a yield of 71%.

[0059] 1 H NMR (300 MHz, CDC13) δ 8.00 (d, J = 7.6 Hz, 2H), 7.54 (d, J = 7.3 Hz, 1H), 7.43 (t, J = 7.7 Hz, 1H), 4.39 (d, J = 5.2 Hz, 2H), 1.47 (s, 9H).

[0060] Preparation of Example 2 intermediate, 4-(N-Boc-aminomethyl)benzoic acid

[0061]

[0062] The preparation method was the same as that of Example 1, except that methyl 4-methylbenzoate was used as the starting material. The target compound was obtained as a white solid with a yield of 65%.

[0063] 1 H NMR (300 MHz, DMSO-d6) δ 12.93 (s, 1H), 7.99-7.69 (m, 2H), 7.45 (q, J = 7.1 Hz, 3H), 4.18 (d, J = 6.1 Hz, 2H), 1.40 (s, 9H).

[0064] Preparation of Example 3 intermediate, 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid

[0065]

[0066] The preparation method refers to example 1, and the target compound is prepared from 5-methyl-2-nitrobenzoic acid methyl ester as a starting material, and is a yellow solid with a yield of 53%.

[0067] 1 H NMR (300 MHz, DMSO-d6) δ 7.96 (d, J = 8.3 Hz, 1H), 7.67 (s, 1H), 7.58 (d, J = 6.7 Hz, 2H), 4.25 (d, J = 6.0 Hz, 2H), 1.40 (s, 9H).

[0068] Preparation of 4-(N-Boc-aminomethyl)-2-nitrobenzoic acid in example 4

[0069]

[0070] The preparation method refers to example 1, and the target compound is prepared from 5-methyl-2-nitrobenzoic acid methyl ester as a starting material, and is a yellow solid with a yield of 53%.

[0071] 1 H NMR (300 MHz, CDCl3) δ 7.84 (d, J = 8.0 Hz, 1H), 7.72 (s, 1H), 7.59 (d, J = 7.9 Hz, 1H), 4.60-4.28 (m, 2H), 1.45 (d, J = 9.6 Hz, 9H).

[0072] Preparation of 2-{[(6S)-1-[3-(aminomethyl)phenyl]-6-(2-methylpropyl)-1,4,7- trioxoboran-2,5-diazepin-7-yl]amino}benzoic acid in example 5

[0073]

[0074] 1, {[(2S)-1-{[2-(methoxycarbonyl)phenyl]amino}-4-methyl-1-oxopentan-2-yl]amino}methane acid-9H-fluoren-9-ylmethyl ester

[0075] To Fmoc-L-leucine (1.0 g, purchased from Bide Pharmatech Co., Ltd.) and 10 mL of dichloromethane, 2 mL of sulfurous dichloride and 1 drop of N,N-dimethylformamide were added at room temperature, and the reaction was refluxed at 40 °C for 6 hours. After cooling to room temperature, it was spin-dried to obtain a viscous liquid.

[0076] Methyl 2-{[(8S)-1-(9H-fluoren-9-yl)-8-(2-methylpropyl)-3,6,9-trioxo-4,7-diaza- 2-oxan-9-yl]amino}benzoate

[0077] 1 H NMR (300 MHz, DMSO-d6) δ 11.26 (s, 1H), 8.47 (d, J = 8.3 Hz, 1H), 8.03 (d, J = 7.2 Hz, 1H), 7.93 (d, J = 7.9 Hz, 1H), 7.86 (d, J = 7.5 Hz, 2H), 7.75 (dd, J = 15.1, 7.4 Hz, 2H), 7.60 (t, J = 7.8 Hz, 1H), 7.38 (t, J = 7.3 Hz, 2H), 7.28 (t, J = 7.2 Hz, 2H), 7.16 (t, J = 7.6 Hz, 1H), 4.54 - 4.20 (m, 3H), 4.09 (dd, J = 14.0, 7.3 Hz, 1H), 3.69 (s, 3H), 1.78 - 1.50 (m, 3H), 0.89 (dd, J = 11.1, 6.0 Hz, 6H).

[0078] 2,2-{[(8S)-1-(9H-fluoren-9-yl)-8-(2-methylpropyl)-3,6,9-trioxo-4,7-diaza-2-oxan- 9-yl]amino}benzoic acid

[0079] Methyl 2-{[(8S)-1-(9H-fluoren-9-yl)-8-(2-methylpropyl)-3,6,9-trioxo-4,7-diaza- 2-oxan-9-yl]amino}benzoate

[0080] Fmoc-Gly (0.6 g, purchased from Bide Pharmatech Co., Ltd.) was dissolved in an appropriate amount of DMF at room temperature, HATU (1.4 g, CAS No.: 148893-10-1) and DIEA (0.5 g) were added, and the reaction was stirred at 0°C for 60 minutes. The above white solid was dissolved in a small amount of DMF and added dropwise to the reaction solution, and after stirring at 0°C for 30 minutes, it was moved to room temperature and reacted overnight. After the reaction was completed, 30 mL of water was added, and the organic phase was extracted with EA three times, combined, washed with water three times, and then dried over anhydrous Na2SO4, dried under reduced pressure, and purified by silica gel column chromatography to obtain the target compound as a white solid with a yield of 48%.

[0081] 1 H NMR (300 MHz, DMSO-d6) δ 11.19 (s, 1H), 8.55 (d, J = 7.1 Hz, 1H), 8.45 (d, J = 8.3 Hz, 1H), 7.95 (d, J = 7.2 Hz, 1H), 7.88 (d, J = 7.4 Hz, 2H), 7.71 (d, J = 7.4 Hz, 2H), 7.62 (t, J = 7.4 Hz, 2H), 7.55 (t, J = 5.9 Hz, 2H), 7.41 (t, J = 7.4 Hz, 2H), 7.31 (t, J = 7.4 Hz, 2H), 7.19 (t, J = 7.6 Hz, 1H), 4.42-4.10 (m, 4H), 3.97-3.73 (m, 5H), 1.86-1.51 (m, 3H), 0.89 (dd, J = 13.4, 5.9 Hz, 6H).

[0082] 3, 2-{[(6S)-1-[3-(2,2-dimethyl-4-oxo-5-aza-3-oxa-hex-6-yl)phenyl]-6-(2- methylpropyl)-1, 4, 7-trioxo-2, 5-diazepin-7-yl]amino}benzoic acid methyl ester

[0083] -16°C, to methyl 2-{[(8S)-1-(9H-fluoren-9-yl)-8-(2-methylpropyl)-3,6,9- trioxo-4, 7-diazepan-9-yl]amino}benzoate (1.0 g) and 20 mL of acetonitrile, DBU (0.3 g) was slowly added dropwise, and after the addition was completed, the reaction was stirred for 30 minutes. After the reaction was completed, it was dried to obtain a white solid.

[0084] To 3-(N-Boc-amino methyl)benzoic acid (0.5 g) dissolved in DMF, HATU (1.4 g) and DIEA (0.5 g) were added at room temperature and stirred for 60 minutes at 0 °C. The white solid was dissolved in a small amount of DMF and added dropwise to the reaction solution. After stirring for 30 minutes at 0 °C, it was allowed to react at room temperature overnight. After the reaction was completed, 30 mL of water was added, and EA was extracted three times. The organic layer was combined, washed with water three times, dried over anhydrous Na2SO4, and dried under reduced pressure to obtain the target compound as a white solid with a yield of 51%.

[0085] 1 H NMR (300 MHz, DMSO-d6) 11.18 (s, 1H), 8.63 (dd, J = 14.3, 6.4 Hz, 2H), 8.43 (d, J = 8.2 Hz, 1H), 7.94 (dd, J = 7.9, 1.2 Hz, 1H), 7.72 (d, J = 8.5 Hz, 2H), 7.62 (dd, J = 11.5, 4.2 Hz, 1H), 7.41 (dd, J = 13.5, 5.8 Hz, 3H), 7.18 (t, J = 7.6 Hz, 1H), 4.31 (q, J = 7.1 Hz, 1H), 4.12 (dd, J = 20.2, 5.8 Hz, 2H), 3.87 (s, 2H), 1.65 (d, J = 6.4 Hz, 3H), 1.37 (s, 2H), 0.92 (d, J = 6.2 Hz, 10H), 0.87 (d, J = 6.1 Hz, 6H).

[0086] 4, 2-{[(6S)-1-[3-(2,2-dimethyl-4-oxo-5-aza-3-oxa-hex-6-yl)phenyl]-6-(2-methylpropyl)- 1,4,7-trioxo-2,5-diazeppt-hept-7-yl]amino}benzoic acid

[0087] To methyl 2-{[(6S)-1-[4-(2,2-dimethyl-4-oxo-5-aza-3-oxa-hex-6-yl)phenyl]-6-(2- methylpropyl)-1,4,7-trioxo-2,5-diazeppt-hept-7-yl]amino}benzoate (1 g) and 10 mL of THF, lithium hydroxide (0.3 g) and 5 mL of H2O were added at room temperature and allowed to react for 2 hours at room temperature. After the reaction was completed, it was acidified with an appropriate amount of acetic acid, 20 mL of water was added, and DCM was extracted three times. The organic phase was combined, dried over anhydrous Na2SO4, and dried under reduced pressure to obtain the target compound as a white solid with a yield of 87%.

[0088] 1H NMR (300 MHz, DMSO-d6) δ 13.77 (s, 1H), 11.72 (s, 1H), 8.63 (m, 3H), 8.02 (d, J = 6.7 Hz, 1H), 7.74 (d, J = 8.3 Hz, 2H), 7.61 (t, J = 7.3 Hz, 1H), 7.43 (dd, J = 6.0 Hz, 3H), 7.17 (t, J = 7.4 Hz, 1H), 4.35 (d, J = 9.5 Hz, 1H), 4.12 (m, 4H), 1.65 (dd, J = 9.1 Hz, 2H), 1.39 (s, 9H), 1.23 (s, 1H), 0.90 (m, 6H).

[0089] 5. 2-{[(6S)-1-[3-(aminomethyl)phenyl]-6-(2-methylpropyl)-1,4,7-trioxo-2,5- diazepan-7-yl]amino}benzoic acid

[0090] To 2-{[(6S)-1-[4-(2,2-dimethyl-4-oxo-5-aza-3-oxahexan-6-yl)phenyl]-6-(2- methylpropyl)-1,4,7-trioxo-2,5-diazepan-7-yl]amino}benzoic acid (0.2 g) and 5 mL DCM was added dropwise 1 mL TFA at room temperature. The reaction was stirred for 30 min and concentrated to dryness to give the target compound as a white solid in 72% yield.

[0091] 1 H NMR (300 MHz, DMSO) δ 12.62 (s, 1H), 9.25 (s, 1H), 8.76 (d, J = 7.6 Hz, 1H), 8.57 (d, J = 8.3 Hz, 3H), 8.23 (s, 1H), 8.08 - 7.99 (m, 1H), 7.93 (d, J = 7.7 Hz, 1H), 7.62 (d, J = 7.5 Hz, 1H), 7.50 (dd, J = 16.2, 8.3 Hz, 2H), 7.09 (t, J = 7.3 Hz, 1H), 4.31 (t, J = 7.6 Hz, 1H), 4.22 - 4.03 (m, 4H), 1.72 - 1.58 (m, 3H), 0.90 (dd, J = 14.7, 5.2 Hz, 6H). MS (ESI): [M+H] + 441 m / z.

[0092] Example 6 Preparation of 2-{[(6S)-1-[4-(aminomethyl)phenyl]-6-(2-methylpropyl)-1,4,7- trioxo-2,5-diazepan-7-yl]amino}benzoic acid

[0093]

[0094] The preparation method refers to example 5, taking Fmoc-L-leucine as the starting material, and 3-(N-Boc-aminomethyl)benzoic acid in step 3 is replaced by 4-(N-Boc- aminomethyl)benzoic acid to obtain the target compound as a white solid with a yield of 65%.

[0095] MS (ESI): [M+H] + 441.5 m / z.

[0096] 1H NMR (300 MHz, DMSO-d6) δ 13.96 (s, 1H), 9.35 (s, 1H), 8.62 (d, J = 7.8 Hz, 2H), 8.53 (d, J = 8.2 Hz, 1H), 8.04 (t, J = 6.5 Hz, 3H), 7.55 (d, J = 8.0 Hz, 2H), 7.33 (t, J = 7.3 Hz, 1H), 7.00 (t, J = 7.4 Hz, 1H), 4.33 (t, J = 7.8 Hz, 1H), 4.13 (s, 4H), 1.65-1.56 (m, 3H), 0.89 (dd, J = 12.4, 5.1 Hz, 6H).

[0097] Example 7 Preparation of 2-{[(3S,6S)-3-(4-aminobutyl)-1-[3- (aminomethyl)phenyl]-6-(2-methylpropyl)-1,4,7-trioxo-2,5-diazepptn-7-yl]amino}benzoic acid

[0098]

[0099] The preparation method refers to example 5, taking Fmoc-L-leucine as the starting material, and Fmoc-glycine in step 2 is replaced by N-Fluorenylmethoxycarbonyl-N'-tert- butyloxycarbonyl-L-lysine (purchased from Bide Pharmatech Co., Ltd.) to obtain the target compound as a white solid with a yield of 71%.

[0100] MS (ESI): [M+H] + 512.6 m / z.

[0101] 1H NMR (300 MHz, DMSO-d6) δ 11.55 (s, 1H), 8.50 (dd, J = 14.4, 7.9 Hz, 3H), 8.39 (s, 2H), 8.00 (d, J = 8.2 Hz, 2H), 7.90 (d, J = 7.8 Hz, 3H), 7.66 - 7.44 (m, 3H), 7.15 (t, J = 7.5 Hz, 1H), 4.58 (t, J = 10.7 Hz, 1H), 4.28 (dd, J = 13.8, 7.3 Hz, 1H), 4.10 (s, 2H), 2.79 (s, 2H), 1.89 (s, 1H), 1.65 (ddd, J = 22.3, 19.8, 9.9 Hz, 6H), 1.40 (d, J = 9.7 Hz, 2H), 0.89 (dd, J = 16.8, 6.1 Hz, 6H).

[0102] Example 8 Preparation of 2-{[(3S,6S)-3-(4-aminobutyl)-1-[4- (aminomethyl)phenyl]-6-(2-methylpropyl)-1,4,7-trioxo-2,5-diazepan-7-yl]amino}benzoic acid

[0103]

[0104] The preparation method refers to Example 5, using Fmoc-L-leucine as the starting material, and Fmoc-glycine in step 2 is replaced by N-Fluorenylmethoxycarbonyl-N'-tert- butyloxycarbonyl-L-lysine (purchased from Bide Pharmatech Co., Ltd.), and 3-(N-Boc- aminomethyl)benzoic acid in step 3 is replaced by 4-(N-Boc-aminomethyl)benzoic acid. The target compound is prepared and obtained as a white solid with a yield of 63%.

[0105] MS (ESI): [M+H] + 512.6 m / z.

[0106] 1 H NMR (300 MHz, DMSO-d6) δ 11.54 (s, 1H), 8.52 (dd, J = 14.4, 6.5 Hz, 3H), 8.39 (s, 2H), 8.00 (d, J = 7.7 Hz, 1H), 7.92 (d, J = 8.0 Hz, 2H), 7.81 (s, 2H), 7.57 (dd, J = 16.7, 8.0 Hz, 3H), 7.15 (t, J = 7.5 Hz, 1H), 4.57 (s, 1H), 4.27 (d, J = 6.4 Hz, 1H), 4.11 (s, 2H), 2.77 (s, 2H), 1.89 (s, 1H), 1.80 - 1.51 (m, 6H), 1.40 (s, 2H), 0.88 (dd, J = 17.6, 5.9 Hz, 6H).

[0107] Preparation of 2-{[(6S,9S)-1-amino-6-({[3-(aminomethyl)phenyl]carbonyl}amino)-1- azanyl-9-(2-methylpropyl)-7,10-dioxo-2,8-diazadec-10-yl]amino}benzoic acid

[0108]

[0109] The preparation method refers to Example 5, taking Fmoc-L-leucine as the starting material, and Fmoc-Pbf-L-arginine (purchased from Bide Pharmaceutical) is used instead of Fmoc-glycine in step 2. The target compound is prepared as a white solid with a yield of 67%.

[0110] MS (ESI): [M+H] + 540.6 m / z.

[0111] 1 H NMR (300 MHz, DMSO-d6) δ 11.97 (s, 1H), 8.56 (d, J = 8.0 Hz, 2H), 8.49 (d, J = 7.6 Hz, 1H), 8.28 (s, 3H), 8.02 (d, J = 6.7 Hz, 2H), 7.92 (d, J = 7.6 Hz, 1H), 7.64 (d, J = 7.5 Hz, 1H), 7.53 (t, J = 7.6 Hz, 2H), 7.35 (s, 2H), 7.13 (t, J = 7.5 Hz, 1H), 4.63 (s, 1H), 4.31 (dd, J = 14.0, 7.0 Hz, 1H), 4.11 (s, 2H), 3.19 (d, J = 4.8 Hz, 2H), 2.04 (dd, J = 31.8, 14.4 Hz, 1H), 1.86 - 1.48 (m, 5H), 1.29 (dd, J = 18.6, 11.5 Hz, 1H), 0.90 (dd, J = 18.3, 6.1 Hz, 6H).

[0112] Example 10 2-{[(6S,9S)-1-amino-6-({[4-(aminomethyl)phenyl]carbonyl}amino)-1- azanyl-9-(2-methylpropyl)-7,10-dioxo-2,8-diazadec-10-yl]amino}benzoic acid

[0113]

[0114] The preparation method refers to example 5, by taking Fmoc-L-leucine as the starting material, Fmoc-Phe is used instead of Fmoc-Gly in step 2, and 4-(N-Boc- aminomethyl)benzoic acid is used instead of 3-(N-Boc-aminomethyl)benzoic acid in step 3, the target compound is prepared as a white solid with a yield of 65%.

[0115] MS (ESI): [M+H] + 540.6 m / z.

[0116] 1 H NMR (300 MHz, DMSO-d6) δ 11.69 (s, 1H), 8.54 (dd, J = 7.9, 4.4 Hz, 2H), 8.47 (d, J = 7.1 Hz, 1H), 8.27 (s, 2H), 8.01 (d, J = 7.9 Hz, 1H), 7.93 (d, J = 8.2 Hz, 2H), 7.56 (t, J = 8.2 Hz, 3H), 7.15 (t, J = 7.5 Hz, 2H), 4.61 (d, J = 3.8 Hz, 1H), 4.31 (dd, J = 13.9, 7.2 Hz, 1H), 4.11 (d, J = 4.3 Hz, 2H), 3.15 (s, 2H), 2.03 - 1.46 (m, 7H), 0.89 (dd, J = 16.5, 6.1 Hz, 6H).

[0117] Example 11 Preparation of 2-{[(6S)-1-[5-(aminomethyl)-2-nitrophenyl]-6-(2- methylpropyl)-1,4,7-trioxo-2,5-diazepan-7-yl]amino}benzoic acid

[0118]

[0119] The preparation method refers to example 5, by taking Fmoc-L-leucine as the starting material, Fmoc-Phe is used instead of Fmoc-Gly in step 2, and 4-(N-Boc- aminomethyl)benzoic acid is used instead of 3-(N-Boc-aminomethyl)benzoic acid in step 3, the target compound is prepared as a white solid with a yield of 65%.

[0120] MS (ESI): [M+H] + 486.6 m / z.

[0121] 1H NMR (300 MHz, DMSO-d6) δ 11.85 (s, 1H), 8.95 (t, J = 5.2 Hz, 1H), 8.74 (d, J = 7.1 Hz, 1H), 8.63 (d, J = 8.3 Hz, 1H), 8.49 (s, 2H), 8.14 (d, J = 8.2 Hz, 1H), 8.04 (d, J = 7.2 Hz, 1H), 7.77 (d, J = 8.3 Hz, 2H), 7.62 (t, J = 7.3 Hz, 1H), 7.18 (t, J = 7.5 Hz, 1H), 4.37 (d, J = 9.8 Hz, 1H), 4.21 (s, 2H), 4.09 (m, 2H), 1.72 (t, J = 9.5 Hz, 2H), 1.48 - 1.09 (m, 1H), 0.94 (dd, J = 14.6, 5.6 Hz, 6H).

[0122] Preparation of 2-{[(6S,9S)-1-amino-6-({[5-(aminomethyl)-2- nitrophenyl]carbonyl}amino)-1-azoniabicyclo[4.4.0]dec-9-yl]amino}benzoic acid

[0123]

[0124] The preparation method is referred to Example 5, and Fmoc-L-leucine is used as the starting material. In step 2, Fmoc-glycine is replaced by Fmoc-Pbf-L-arginine, and in step 3, 3-(N-Boc- aminomethyl)benzoic acid is replaced by 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid. The target compound is prepared and is a white solid with a yield of 72%.

[0125] MS (ESI): [M+H] + 585.6 m / z.

[0126] 1H NMR (300 MHz, DMSO-d6) δ 12.10 (s, 1H), 8.84 (d, J = 7.7 Hz, 1H), 8.59 - 8.55 (m, 2H), 8.41 (s, 3H), 8.12 (d, J = 8.4 Hz, 1H), 8.02 (dd, J = 7.9, 1.4 Hz, 1H), 7.76 (d, J = 8.5 Hz, 1H), 7.70 (s, 1H), 7.52 (t, J = 7.2 Hz, 1H), 7.36 (s, 3H), 7.12 (t, J = 7.6 Hz, 1H), 4.61 (s, 1H), 4.32 (dd, J = 14.4, 7.1 Hz, 1H), 4.20 (s, 2H), 3.22 (d, J = 4.6 Hz, 2H), 2.08 - 1.50 (m, 7H), 0.92 (dd, J = 15.0, 6.4 Hz, 6H).

[0127] Example 13 Preparation of 2-{[(3S,6S)-3-(4-aminobutyl)-1-[5-(aminomethyl)-2- nitrophenyl]-6-(2-methylpropyl)-1,4,7-trioxo-2,5-diazeppt-heptyl]amino}benzoic acid

[0128]

[0129] The preparation method refers to Example 5, using Fmoc-L-leucine as the starting material, and corresponding N-fluorenylmethoxycarbonyl-N'-tert-butoxycarbonyl-L-lysine is used for Fmoc-glycine in step 2, and 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid is used for 3-(N-Boc-aminomethyl)benzoic acid in step 3. The target compound is prepared and is a white solid with a yield of 63%.

[0130] MS (ESI): [M+H] + 557.6 m / z.

[0131] 1H NMR (300 MHz, DMSO-d6) δ 11.54 (s, 1H), 8.82 (d, J = 7.9 Hz, 1H), 8.61 - 8.46 (m, 3), 8.13 (d, J = 8.4 Hz, 1H), 8.01 (d, J = 7.9 Hz, 1H), 7.86 (s, 2H), 7.78 - 7.66 (m, 2H), 7.60 (t, J = 7.3 Hz, 1H), 7.17 (t, J = 7.4 Hz, 1H), 4.66 - 4.46 (m, 1H), 4.31 (dd, J = 13.7, 7.3 Hz, 1H), 4.21 (s, 2H), 2.78 (d, J = 4.7 Hz, 2H), 1.92 - 1.52 (m, 7H), 1.44 (s, 2H), 0.93 (dd, J = 13.7, 6.3 Hz, 6H).

[0132] Example 14 Preparation of 2-{[(3S,6S)-3-(4-aminobutyl)-1-[5-(aminomethyl)-2- [(1-oxyltetradecyl)amino]phenyl]-6-(2-methylpropyl)-1,4,7-trioxa-2,5- diazepan-7-yl]amino}benzoic acid

[0133]

[0134] The preparation method is referred to Example 5, using Fmoc-L-leucine as the starting material, and Fmoc-glycine in step 2 is replaced by N-fluorenylmethoxycarbonyl-N'- tert-butyloxycarbonyl-L-lysine, and 3-(N-Boc-aminomethyl)benzoic acid in step 3 is replaced by 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid. After step 3, Pd / C and H2 are used for reduction, the reaction is completed, and then spin dry, dissolve in THF, add DIEA, slowly drop myristoyl chloride at 0°C, purify the pure product by silica gel column chromatography after the reaction is completed, and continue steps 4 and 5 to obtain the target compound as a white solid with a yield of 69%.

[0135] MS (ESI): [M+H] + 737.8 m / z.

[0136] 1H NMR (300 MHz, DMSO-d6) δ 11.68 (s, 1H), 10.80 (s, 1H), 8.67 (d, J = 6.5 Hz, 1H), 8.55 (d, J = 8.3 Hz, 1H), 8.35 (d, J = 8.5 Hz, 2H), 8.01 (d, J = 7.8 Hz, 1H), 7.87 (s, 2H), 7.79 (s, 1H), 7.56 (dd, J = 11.7, 6.3 Hz, 2H), 7.15 (t, J = 7.4 Hz, 1H), 4.53 (s, 1H), 4.37 - 4.27 (m, 1H), 4.03 (s, 2H), 2.79 (s, 2H), 2.31 (t, J = 7.3 Hz, 2H), 1.62 (m, 11H), 1.21 (d, J = 3.0 Hz, 22H), 0.91 (dd, J = 16.1, 6.2 Hz, 6H), 0.84 (d, J = 6.7 Hz, 3H).

[0137] Example 15 Preparation of 2-{[(6S)-1-[5-(aminomethyl)-2-[(1-oxyltetradecyl)amino]phenyl]-6-(2- methylpropyl)-1,4,7-trioxyl-2,5-diazepan-7-yl]amino}benzoic acid

[0138]

[0139] The preparation method refers to Example 5, and Fmoc-L-leucine is used as the starting material. In step 3, 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid is used instead of 3-(N-Boc- aminomethyl)benzoic acid. After step 3, Pd / C and H2 are used for reduction. After the reaction is completed, rotary evaporation is performed, the product is dissolved in THF, DIEA is added, and myristoyl chloride is slowly added dropwise at 0°C. After the reaction is completed, the product is purified by silica gel column chromatography. The above steps 4 and 5 are continued to obtain the target compound, which is a white solid, and the yield is 71%.

[0140] MS (ESI): [M+H] + 666.7 m / z.

[0141] 1 H NMR (300 MHz, DMSO-d6) δ 12.14 (s, 1H), 11.15 (s, 1H), 9.14 (d, J = 13.7 Hz, 2H), 8.90 (s, 1H), 8.60 (m, 4H), 8.30 (s, 1H), 8.08 (s, 1H), 7.84 (s, 2H), 7.43 (s, 1H), 4.63 (s, 1H), 4.34 (m, 4H), 2.58 (s, 2H), 1.98 (s, 3H), 1.81 (s, 2H), 1.49 (s, 20H), 1.17 (s, 9H).

[0142] Preparation of 2-{[(6S,9S)-1-amino-6-({[5-(aminomethyl)-2-[(1-oxyltetradecyl)amino] phenyl]carbonyl}amino)-1-oxyl-9-(2-methylpropyl)-7,10-dioxo-2,8-diazadecyl]amino} benzoic acid

[0143]

[0144] The preparation method is referred to Example 5, using Fmoc-L-leucine as the starting material, Fmoc-Pbf-L-arginine is used instead of Fmoc-glycine in step 2, and 5-(N-Boc- aminomethyl)-2-nitrobenzoic acid is used instead of 3-(N-Boc-aminomethyl)benzoic acid in step 3. After step 3, Pd / C and H2 are used for reduction, after the reaction is completed, rotary evaporation is performed, THF is used for dissolution, DIEA is added, myristoyl chloride is slowly added dropwise at 0°C, after the reaction is completed, silica gel column chromatography is used for purification to obtain a pure product, and the above steps 4 and 5 are continued to obtain the target compound, which is a white solid with a yield of 57%.

[0145] MS (ESI): [M+H] + 765.8 m / z.

[0146] 1 H NMR (300 MHz, DMSO-d6) δ 10.86 (s, 1H), 8.66 (t, J = 6.9 Hz, 2H), 8.59 (d, J = 8.3 Hz, 1H), 8.37 (d, J = 8.5 Hz, 1H), 8.22 (s, 3H), 8.03 (d, J = 7.8 Hz, 1H), 7.85 (s, 1H), 7.57 (d, J = 8.6 Hz, 1H), 7.49 (t, J = 7.6 Hz, 1H), 7.39 (s, 2H), 7.10 (t, J = 7.5 Hz, 1H), 4.57 (s, 1H), 4.36 (dd, J = 14.6, 7.3 Hz, 1H), 4.04 (s, 2H), 3.23 (d, J = 4.4 Hz, 2H), 2.32 (t, J = 7.1 Hz, 2H), 2.17 (dd, J = 16.9, 9.5 Hz, 1H), 1.81 (dd, J = 13.2, 6.5 Hz, 2H), 1.73 - 1.44 (m, 6H), 1.23 (s, 21H), 0.94 (d, J = 6.3 Hz, 3H), 0.86 (dd, J = 9.6, 6.5 Hz, 6H).

[0147] Preparation of Example 17 2-{[(6S)-1-[4-(aminomethyl)-2-[(1-oxosuberyl)amino]phenyl]-6-(2- methylpropyl)-1,4,7-trioxo-2,5-diazepptan-7-yl]amino}benzoic acid

[0148]

[0149] The preparation method is referred to Example 5, using Fmoc-L-leucine as the starting material, and 4-(N-Boc-aminomethyl)-2-nitrobenzoic acid is used instead of 3-(N-Boc- aminomethyl)benzoic acid in step 3. After step 3, Pd / C and H2 are used for reduction, and after the reaction is completed, rotary evaporation is performed, and then the product is dissolved in THF, DIEA is added, and myristoyl chloride is slowly added dropwise at 0°C. After the reaction is completed, the product is purified by silica gel column chromatography, and the above steps 4 and 5 are continued to obtain the target compound as a white solid with a yield of 69%.

[0150] MS (ESI): [M+H] + 666.7 m / z.

[0151] 1 H NMR (300 MHz, DMSO-d6) δ 11.95 (s, 1H), 11.18 (s, 1H), 9.08 (s, 1H), 8.76 (d, J = 7.2 Hz, 1H), 8.60 (d, J = 8.1 Hz, 1H), 8.50 (s, 1H), 8.38 (s, 2H), 8.01 (d, J = 7.5 Hz, 1H), 7.80 (d, J = 8.0 Hz, 1H), 7.56 (t, J = 7.4 Hz, 1H), 7.22 (d, J = 7.7 Hz, 1H), 7.13 (t, J = 7.4 Hz, 1H), 4.34 (d, J = 4.0 Hz, 1H), 4.05 (s, 4H), 2.30 (t, J = 7.2 Hz, 2H), 1.71 (d, J = 9.4 Hz, 3H), 1.55 (s, 2H), 1.21 (s, 20H), 0.91 (dd, J = 13.5, 5.7 Hz, 6H), 0.84 (t, J = 6.6 Hz, 3H).

[0152] Preparation of Example 18 2-{[(6S)-1-[4-(aminomethyl)-2-[(1-oxodecyl)amino]phenyl]-6-(2- methylpropyl)-1,4,7-trioxo-2,5-diazepptan-7-yl]amino}benzoic acid

[0153]

[0154] The preparation method refers to example 5, using Fmoc-L-leucine as the starting material, and 3-(N-Boc-aminomethyl)benzoic acid in step 3 is replaced by 4-(N-Boc-aminomethyl)-2-nitrobenzoic acid. After step 3, reduction is performed with Pd / C and H2, after the reaction is completed, spin dry, dissolve in THF, then add DIEA, slowly drop lauroyl chloride at 0°C, after the reaction is completed, purify the pure product by silica gel column chromatography, continue the above steps 4, 5 to obtain the target compound, which is a white solid, with a yield of 59%.

[0155] MS (ESI): [M+H] + 632.7 m / z.

[0156] 1 H NMR (300 MHz, DMSO-d6) δ 11.62 (s, 1H), 11.01 (s, 1H), 8.90 (s, 1H), 8.65 (d, J = 7.1 Hz, 1H), 8.47 (d, J = 8.3 Hz, 1H), 8.37 (s, 1H), 8.23 (s, 2H), 7.87 (d, J = 7.3 Hz, 1H), 7.62 (t, J = 7.1 Hz, 1H), 7.45 (t, J = 7.6 Hz, 1H), 7.14-6.92 (m, 2H), 4.19 (dd, J = 13.5, 7.1 Hz, 1H), 4.09-3.82 (m, 4H), 2.15 (t, J = 7.2 Hz, 2H), 1.58 (dd, J = 23.6, 5.7 Hz, 3H), 1.41 (s, 2H), 1.07 (s, 12H), 0.76 (dd, J = 13.6, 5.9 Hz, 6H), 0.69 (t, J = 5.7 Hz, 3H).

[0157] Example 19 Preparation of 2-{[(6S)-1-[4-(aminomethyl)-2-[(1-oxyl dodecyl)amino]phenyl]-6-(2-methylpropyl)-1,4,7-trioxo-2,5-diazepan-7-yl]amino}benzoic acid

[0158]

[0159] The preparation method refers to example 5, using Fmoc-L-leucine as the starting material, and 3-(N-Boc-aminomethyl)benzoic acid in step 3 is replaced by 4-(N-Boc-aminomethyl)-2-nitrobenzoic acid. After step 3, reduction is performed with Pd / C and H2, after the reaction is completed, spin dry, dissolve in THF, then add DIEA, slowly drop lauroyl chloride at 0°C, after the reaction is completed, purify the pure product by silica gel column chromatography, continue the above steps 4, 5 to obtain the target compound, which is a white solid, with a yield of 59%.

[0160] MS (ESI): [M+H] + 638.7 m / z.

[0161] 1 H NMR (300 MHz, DMSO-d6) δ 11.77 (s, 1H), 11.18 (s, 1H), 9.05 (t, J = 5.4 Hz, 1H), 8.80 (d, J = 7.1 Hz, 1H), 8.64 (d, J = 8.3 Hz, 1H), 8.54 (s, 1H), 8.41 (s, 2H), 8.04 (d, J = 7.5 Hz, 1H), 7.81 (d, J = 8.1 Hz, 1H), 7.61 (t, J = 7.7 Hz, 1H), 7.25 (d, J = 8.0 Hz, 1H), 7.17 (t, J = 7.6 Hz, 1H), 4.46 - 4.28 (m, 1H), 4.30 - 3.93 (m, 4H), 2.32 (t, J = 7.2 Hz, 2H), 1.76 (m, 3H), 1.58 (s, 2H), 1.24 (s, 16H), 0.94 (dd, J = 13.2, 5.9 Hz, 6H), 0.86 (t, J = 6.4 Hz, 3H).

[0162] Example 20 Preparation of 2-{[(6S)-1-[4-(aminomethyl)-2-[(1-oxylhexadecyl)amino]phenyl]-6-(2- methylpropyl)-1,4,7-trioxa-2,5-diazepan-7-yl]amino}benzoic acid

[0163]

[0164] The preparation method refers to Example 5, and Fmoc-L-leucine is used as the starting material. In step 3, 3-(N-Boc-aminomethyl)benzoic acid is replaced by 4-(N-Boc- aminomethyl)-2-nitrobenzoic acid. After step 3, Pd / C and H2 are used for reduction, the reaction is completed, then the reaction system is rotary evaporated, dissolved in THF, DIEA is added, palmitoyl chloride is slowly added dropwise at 0°C, after the reaction is completed, the pure product is purified by silica gel column chromatography, and the target compound is prepared by continuing steps 4 and 5, and the target compound is a white solid with a yield of 62%.

[0165] MS (ESI): [M+H] + 694.8 m / z.

[0166] 1H NMR (300 MHz, DMSO-d6) δ 11.78 (s, 1H), 11.18 (s, 1H), 9.05 (s, 1H), 8.79 (d, J = 6.8 Hz, 1H), 8.63 (d, J = 8.1 Hz, 1H), 8.53 (s, 1H), 8.39 (s, 2H), 8.03 (d, J = 7.3 Hz, 1H), 7.80 (d, J = 7.9 Hz, 1H), 7.61 (t, J = 7.7 Hz, 1H), 7.28 - 7.09 (m, 2H), 4.36 (s, 1H), 4.28 - 3.92 (m, 4H), 2.31 (d, J = 6.6 Hz, 2H), 1.72 (s, 3H), 1.57 (s, 3H), 1.24 (s, 24H), 0.93 (dd, J = 13.2, 5.3 Hz, 6H), 0.88 (s, 3H).

[0167] Preparation of 2-{[(6S)-1-[5-(aminomethyl)-2-[(1-oxyldecyl)amino]phenyl]-6-(2- methylpropyl)-1,4,7-trioxo-2,5-diazepan-7-yl]amino}benzoic acid

[0168]

[0169] The preparation method refers to Example 5, using Fmoc-L-leucine as the starting material, and 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid is used instead of 3-(N-Boc- aminomethyl)benzoic acid in step 3. After step 3, reduction is performed with Pd / C and H2. After the reaction is completed, spin dry, dissolve in THF, then add DIEA, slowly drop decanoyl chloride at 0°C, and after the reaction is completed, purify the pure product by silica gel column chromatography. Continue with steps 4 and 5 described above to obtain the target compound as a white solid with a yield of 65%.

[0170] MS (ESI): [M+Na] + 633.7 m / z.

[0171] 1H NMR (300 MHz, DMSO-d6) δ 11.99 (s, 1H), 10.99 (s, 1H), 9.03 (s, 1H), 8.88 (d, J = 7.2 Hz, 1H), 8.65 (d, J = 8.2 Hz, 1H), 8.43 (d, J = 8.4 Hz, 3H), 8.06 (d, J = 6.8 Hz, 1H), 7.92 (s, 1H), 7.62 (t, J = 7.8 Hz, 2H), 7.19 (t, J = 7.4 Hz, 1H), 4.42 (d, J = 9.7 Hz, 1H), 4.28 - 4.11 (m, 2H), 4.06 (s, 2H), 2.35 (t, J = 7.3 Hz, 2H), 1.83 - 1.66 (m, 3H), 1.58 (s, 2H), 1.26 (s, 12H), 0.96 (dd, J = 14.0, 5.7 Hz, 6H), 0.88 (t, J = 6.5 Hz, 3H).

[0172] Example 22 Preparation of 2-{[(6S)-1-[5-(aminomethyl)-2-[(1-oxyl dodecyl)amino]phenyl]-6-(2-methylpropyl)-1,4,7-trioxa-2,5-diazepan-7-yl]amino}benzoic acid

[0173]

[0174] The preparation method refers to Example 5, using Fmoc-L-leucine as the starting material, and 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid is used instead of 3-(N-Boc- aminomethyl)benzoic acid in step 3. After step 3, Pd / C and H2 are used for reduction, the reaction is completed, and then rotary evaporation is performed. The residue is dissolved in THF, and then DIEA is added. Dodecanoyl chloride is slowly added dropwise at 0°C. After the reaction is completed, the pure product is purified by silica gel column chromatography. The above steps 4 and 5 are continued to obtain the target compound, which is a white solid, and the yield is 63%.

[0175] MS (ESI): [M+H] + 638.7 m / z.

[0176] 1H NMR (300 MHz, DMSO-d6) δ 11.94 (s, 1H), 10.94 (s, 1H), 8.97 (s, 1H), 8.86 (d, J = 6.6 Hz, 1H), 8.63 (d, J = 8.2 Hz, 1H), 8.40 (d, J = 8.5 Hz, 4H), 8.04 (d, J = 7.3 Hz, 1H), 7.87 (s, 1H), 7.58 (d, J = 7.9 Hz, 2H), 7.17 (t, J = 7.1 Hz, 1H), 4.38 (s, 1H), 4.15 (m, 2H), 4.04 (s, 2H), 2.33 (s, 2H), 1.73 (s, 3H), 1.56 (s, 2H), 1.24 (s, 16H), 0.94 (dd, J = 13.7, 4.2 Hz, 6H), 0.87 (s, 3H).

[0177] Example 23 Preparation of 2-{[(6S)-1-[5-(aminomethyl)-2-[(1-oxylhexadecyl)amino]phenyl]-6-(2- methylpropyl)-1,4,7-trioxyl-2,5-diazepptan-7-yl]amino}benzoic acid

[0178]

[0179] The preparation method refers to Example 5, using Fmoc-L-leucine as the starting material, and 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid is used instead of 3-(N-Boc- aminomethyl)benzoic acid in step 3. After step 3, reduction is performed with Pd / C and H2, the reaction is completed, then spin-drying is performed, dissolution is performed with THF, DIEA is added, palmitoyl chloride is slowly added dropwise at 0°C, after the reaction is completed, the pure product is purified by silica gel column chromatography, and the above steps 4 and 5 are continuously performed to obtain the target compound, which is a white solid, and the yield is 68%.

[0180] MS (ESI): [M+H] + 694.7 m / z.

[0181] 1H NMR (300 MHz, DMSO-d6) δ 12.12 (s, 1H), 11.01 (s, 1H), 9.08 (s, 1H), 8.84 (d, J = 7.2 Hz, 1H), 8.62 (d, J = 8.3 Hz, 1H), 8.41 (d, J = 8.5 Hz, 4H), 8.04 (d, J = 7.5 Hz, 1H), 7.89 (s, 1H), 7.56 (d, J = 7.1 Hz, 2H), 7.15 (t, J = 7.5 Hz, 1H), 4.38 (s, 1H), 4.23 - 4.11 (m, 2H), 4.04 (s, 2H), 2.32 (t, J = 7.1 Hz, 2H), 1.74 (d, J = 11.8 Hz, 3H), 1.56 (s, 2H), 1.24 (s, 24H), 0.93 (dd, J = 13.8, 5.2 Hz, 6H), 0.86 (t, J = 6.4 Hz, 3H).

[0182] Example 24 Preparation of 3-{[(6S)-1-[5-(aminomethyl)-2-[(1-oxyl dodecyl)amino]phenyl]-6-(2-methylpropyl)-1,4,7-trioxa-2,5-diazepan-7-yl]amino}pyridine-4-carboxylic acid

[0183]

[0184] The preparation method is referred to Example 5, and Fmoc-L-leucine is used as the starting material. In step 1, methyl 2-aminobenzoate is replaced by methyl 3-aminoisonicotinate, and in step 3, 3-(N-Boc-aminomethyl)benzoic acid is replaced by 5-(N-Boc-aminomethyl)-2-nitrobenzoic acid. After step 3, Pd / C and H2 are used for reduction. After the reaction is completed, rotary evaporation is performed, and then the residue is dissolved in THF. Then, DIEA is added, and dodecanoyl chloride is slowly added dropwise at 0°C. After the reaction is completed, the product is purified by silica gel column chromatography. Then, steps 4 and 5 are continued to obtain the target compound, which is a white solid, and the yield is 62%.

[0185] MS (ESI): [M+H] + 639.7 m / z.

[0186] 1H NMR (300 MHz, DMSO-d6) δ 11.72 (s, 1H), 10.95 (s, 1H), 9.69 (s, 1H), 9.02 (s, 1H), 8.87 (s, 1H), 8.37 (m, 4H), 7.84 (s, 2H), 7.54 (d, J = 8.0 Hz, 1H), 4.40 (s, 1H), 4.07 (m, 4H), 2.30 (s, 2H), 1.70 (s, 3H), 1.53 (s, 2H), 1.20 (s, 16H), 1.01 - 0.69 (m, 9H).

[0187] Example 25: Biological activity (minimum inhibitory concentration test)

[0188] The minimum inhibitory concentration of the compound on different strains was detected by using microdilution method to evaluate the antibacterial activity of the compound.

[0189] The broth dilution method is one of the earliest used bacterial drug sensitivity determination methods, which can be divided into constant broth dilution method and micro-broth dilution method. The basic principles of the two methods are the same. A certain concentration of antibacterial drugs is used for serial dilution with culture solution containing the bacteria to be tested, and after incubation at a suitable temperature, the minimum drug concentration contained in the test tube without bacterial growth is observed by naked eye as the minimum inhibitory concentration (MIC).

[0190] Experimental steps:

[0191] 1. Preparation of bacterial suspension:

[0192] (1) Bacterial liquid culture: Take 10 μL of the bacteria to be tested from the storage solution and add 1 mL of MH broth (which can be adjusted according to actual needs), and incubate at 37°C in an incubator for 12 hours or so.

[0193] (2) OD600 value determination: The OD value is determined by using a UV spectrophotometer. The bacterial liquid concentration is adjusted to make the OD600 value = 0.1, at which time the bacterial liquid concentration is about 10 8 cfu / mL (about 7-10 times dilution of the culture liquid is required);

[0194] (3) Sample bacterial liquid dilution: The bacteria to be tested are diluted 1000 times based on the dilution factor obtained in step ②, at which time the bacterial liquid concentration is about 10 5 cfu / mL, which is the sample bacterial suspension at this time;

[0195] Note: The bacterial liquid required for determining the OD value should be sampled aseptically in a clean bench, and the remaining bacterial liquid needs to be tested.

[0196] 2. Preparation of antibacterial drugs:

[0197] Antibiotic mother liquor preparation: according to the CLSI standard, the corresponding R (drug resistance) value of the antibacterial drug to be tested is prepared (the mother liquor concentration is much larger than the R value, at least 160 times), and is divided into sterile small tubes and stored at -20°C for standby.

[0198] Note: sterile operation, the diluent of the antibacterial drug needs to be sterilized, and should be filtered (filter membrane pore size is 0.22 μm) after dissolution.

[0199] 3. Operation of drug sensitivity test:

[0200] (1) The antibacterial drug to be tested is diluted by 10 times;

[0201] (2) 100 μL of sterilized MH broth is added to the first to eleventh columns of the sterile 96-well plate (one drug per plate);

[0202] (3) 100 μL of 10-fold diluted drug solution is added to the first column of the sterile 96-well plate, and is sequentially diluted by 10 times to the eleventh column (the final volume of liquid in each well is 100 μL);

[0203] (4) 100 μL of the bacteria to be tested is added to each well of the sterile 96-well plate, and the final volume of liquid in each well is 200 μL (since the whole plate is one kind of drug, each row of the 96-well plate can perform drug sensitivity test of one bacterium, in order to ensure the reliability of the experiment, each strain is repeated 1-2 times, that is, 2-3 rows are performed for one strain, and one plate can perform drug sensitivity test of 2-4 strains of bacteria);

[0204] (5) 200 μL of sterilized MH broth is added to the four wells in the twelfth column of the sterile 96-well plate as negative control, and 200 μL of bacteria solution is added to the four wells below the twelfth column as positive control;

[0205] (6) After the drug and bacteria solution are loaded, the plate cover is covered, and the plate is placed in a 37°C incubator for 18-22 hours to observe the results (the result interpretation refers to the CLSI antibacterial drug sensitivity test interpretation standard).

[0206] Table 1 results of biological activity experiment

[0207]

[0208] The results show that the above compounds have good bacteriostatic effect on bacteria, and examples 14, 15, 16 and 21 are the best. Examples 14, 15, 16 and 21 have better inhibition effect on Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium than other strains, and the bacteriostatic effect is higher or close to the positive drug ampicillin.

[0209] Therefore, the compound of general formula I and its pharmaceutically acceptable salt prepared by the present application has excellent type I signal peptide enzyme inhibition effect. The compound can inhibit Sec and Tat secretion system by inhibiting type I signal peptide enzyme of bacteria, so that the bacteria cannot release mature secretory protein, and thus plays a role in inhibiting bacteria. Therefore, the above-mentioned compound can be used for preparing a drug for preventing, treating or improving bacterial infection. For example, the compound can be used for preparing a drug for treating diseases caused by bacterial infection such as Staphylococcus aureus, Enterococcus faecalis, Streptococcus pyogenes, Escherichia coli and the like.

[0210] As described above, although the present application has been indicated and expressed with reference to specific preferred embodiments, it should not be construed as a limitation on the present application itself. Various changes can be made in form and details without departing from the spirit and scope of the present application defined by the appended claims.

Claims

1. A 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof, characterized in that... Selected from:

2. A 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof, characterized in that... Selected from:

3. A 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof, characterized in that... Selected from:

4. A 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof, characterized in that... Selected from:

5. A 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof, characterized in that... Selected from:

6. A 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof, characterized in that... Selected from:

7. The 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, characterized in that, The pharmaceutically acceptable salts include acid addition salts formed by the compound with the following acids: hydrochloric acid, sulfuric acid, phosphoric acid, hydrobromic acid, acetic acid, trifluoroacetic acid, pyruvic acid, citric acid, tartaric acid, lactic acid, maleic acid, benzenesulfonic acid, succinic acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, fumaric acid, salicylic acid, phenylacetic acid, or mandelic acid; and also include acidic salts formed by the compound with inorganic bases or organic salts made from basic amines.

8. A method for preparing the compound according to any one of claims 1-6, characterized in that, Includes the following steps: Step 1: Compound II reacts with methyl 2-aminobenzoate or methyl 3-aminoisonicotinic acid to obtain compound III; Step 2: Compound III reacts in the presence of a condensing agent to give compound IV; Step 3: Compound IV reacts with an intermediate in the presence of a condensing agent to obtain compound V; Step 4; Compound V is reduced and then reacted with acyl chlorides of different carbon chain lengths to obtain compound VI; Step 5: Compound VI reacts under acidic or basic conditions to yield compound VII; Step Six: Compound VII reacts in the presence of a condensing agent to obtain the target compound; in, R 1 Selected from R 2 Selected from aminomethyl; R 3 Selected from hydrogen, R 4 Selected from isobutyl; R 5 Selected from carboxyl groups; R 6 Selected from hydrogen; Ar is a benzene ring; n is a natural number less than 20.

9. The method for preparing the compound according to claim 8, characterized in that, The method for preparing the intermediate includes the following steps: Step 1: Compound VIII reacts with NBS to give compound IX; Step 2: Compound IX undergoes the Gabriel reaction, with the amino group protected by tert-butyloxycarbonyl to yield compound X; Step 3: Compound X reacts under acidic or basic conditions to obtain an intermediate.

10. A pharmaceutical composition, characterized in that, This includes the 2-(2-(2-benzoylamino)acetamide)benzoic acid compound or a pharmaceutically acceptable salt thereof as described in any one of claims 1-7, and a pharmaceutically acceptable carrier.

11. The pharmaceutical composition according to claim 10, characterized in that, The dosage form of the pharmaceutical composition is tablets, capsules, pills, soft capsules, oral liquids, suspensions, or injections.

12. The use of the compound of claim 1 in the preparation of a medicament for treating bacterial infections, characterized in that, The bacteria are Streptococcus pyogenes, Staphylococcus aureus, Enterococcus faecalis, or Enterococcus faecium.

13. The use of the compound of claim 2 in the preparation of a medicament for treating bacterial infections, characterized in that, The bacteria are Staphylococcus aureus, Enterococcus faecalis, or Enterococcus faecium.

14. The use of the compound of claim 3 in the preparation of a medicament for treating bacterial infections, characterized in that, The bacteria in question is Staphylococcus aureus.

15. The use of the compound of claim 4 in the preparation of a medicament for treating bacterial infections, characterized in that, The bacteria in question are Enterococcus faecalis.

16. The use of the compound of claim 5 in the preparation of a medicament for treating bacterial infections, characterized in that, The bacteria are Staphylococcus aureus, Enterococcus faecalis, Enterococcus faecium, or Streptococcus suis.

17. Use of the compound of claim 6 in the preparation of a medicament for treating bacterial infections, characterized in that, The bacteria in question are Enterococcus faecalis.

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