Glycopeptide compounds having activity of resisting drug-resistant bacteria, and preparation method and application thereof
11696937 · 2023-07-11
Assignee
- Shanghai LaiYi Center For Biopharmaceutical R&D Co., Ltd. (Zhejiang, CN)
- Zhejiang Medicine Co., Ltd. (Zhejiang, CN)
Inventors
- Chang Shao (Shanghai, CN)
- Mei Ge (Shanghai, CN)
- Lingao Ruan (Shanghai, CN)
- Wei Wei (Shanghai, CN)
- Xing Xia (Shanghai, CN)
- Min Rao (Shanghai, CN)
- Qingqian Meng (Shanghai, CN)
- Minyu Luo (Shanghai, CN)
Cpc classification
A61K9/0019
HUMAN NECESSITIES
A61K38/14
HUMAN NECESSITIES
C07K9/008
CHEMISTRY; METALLURGY
International classification
A61K38/14
HUMAN NECESSITIES
A61K9/00
HUMAN NECESSITIES
Abstract
The present invention discloses glycopeptide compounds having activity of resisting drug-resistant bacteria, conforming to glycopeptide compounds represented by general formula (I), ##STR00001## The present invention also provides a preparation method for and an application of the glycopeptide compounds. Upon testing, compared with a second-generation glycopeptide drug oritavancin, the glycopeptide antibiotic compounds have higher inhibition activity on drug-resistant bacterial strains, especially MRSA or VRE. Further testing shows that most of the glycopeptide compounds have safety higher than that of oritavancin and can be prepared into drugs for treating or preventing diseases caused by various bacterial infections, such as skin and soft tissue infections, meningitis, sepsis, pneumonia, arthritis, peritonitis, bronchitis, and empyema.
Claims
1. A glycopeptide compound having activity of resisting drug-resistant bacteria of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE), characterized in that, conforming glycopeptide compounds shown in general formula I: ##STR00031## or a pharmaceutically acceptable salt thereof, wherein: R represents by the following formula: -A-D-E-G; wherein: A is a benzene ring; D is —O— or —NH—; E is —(CH.sub.2)m-, wherein m is 1 to 3; G is the structural formula: ##STR00032## wherein L is any one of halogen, trifluoromethyl, and methoxy.
2. The glycopeptide compounds according to claim 1, wherein R is selected from: 4-(4′-chlorophenylmethoxy)phenyl, 4-(4′-fluorophenylethoxy)phenyl, 4-(4′-chlorophenylethoxy)phenyl, 4-(4′-bromophenylethoxy)phenyl, 4-(3′-bromophenylethoxy)phenyl, 4-(4′-trifluoromethylphenylethoxy)phenyl, 4-(4′-methoxyphenylethoxy)phenyl, 4-(4′-chlorophenylpropoxy)phenyl, 4-(4′-chlorobenzylamino)phenyl, 4-(4′-chlorophenylethylamino)phenyl, 4-(4′-trifluoromethylphenylethylamino)phenyl, and 4-(4′-chloramphetamine)phenyl.
Description
DETAILED DESCRIPTION OF THE INVENTION
(1) The present invention will be further described below in conjunction with specific embodiments. It should be understood that the following examples are only used to illustrate the present invention and not to limit the scope of the present invention.
(2) In the present invention, the following abbreviations have the following meanings. Undefined abbreviations have their generally accepted meanings, unless otherwise stated, all of room temperature are referred to as a temperature of 20° C.-30° C.
(3) DIEA: N,N-diisopropylethylamine
(4) DMF: N,N-dimethylformamide
(5) DMSO: dimethyl sulfoxide
(6) ESI: electrospray ionization mass spectrometry
(7) Fmoc: 9-fluorenyl methoxycarbonyl
(8) H: hour
(9) LD50: median lethal dose
(10) MRSA: Methicillin-resistant Staphylococcus aureus
(11) MIC: minimum inhibitory concentration
(12) MS: mass spectrum
(13) TFA: trifluoroacetic acid
(14) VRE: Vancomycin-resistant Enterococcus
(15) In the present invention, the method for obtaining the compound of general formula II refers to the patent application document of the Chinese patent publication No. CN101928331A. Other raw materials, materials and devices of the present invention are all commercially available.
(16) In the following examples, reverse-phase polymer fillers Uni PS25-300 and Uni PSA30-300 are used for purifying synthesized crude products. After dissolving crude product into methanol (or acetonitrile) aqueous solution, load the sample onto a glass chromatography column with filler at a flow rate of 1 times column volume/h. After loading sample, pre-wash with methanol (or acetonitrile) aqueous solution for 1 hour, and then elute with methanol (or acetonitrile) aqueous solution containing TFA at a flow rate of 1.5 times column volume/h. After eluting 1 times column volume, start with collecting the eluent, concentrate and dry the eluent in order to obtain pure products of each sample.
(17) In the following examples, the ratio of the eluent is referred to as a volume percentage, and the yield is referred to as a molar yield, unless otherwise specified.
(18) The structure of each compounds of the following examples is shown in Table 1.
(19) TABLE-US-00001 TABLE 1 Structures of Each Compounds MS Number R Molecular formula (ESI) 1
Example 1. Preparation of Compound 1
(20) Mix compound of general formula II (0.5 g, 0.3 mmol) with 10 mL DMF-methanol (1:1, v/v), add DIEA (0.1 mL, 0.6 mmol) and 4-phenylmethoxybenzaldehyde (0.085 g, 0.4 mmol)), stir at 65° C. for 2h and then cool to room temperature; add TFA (0.07 mL, 0.9 mmol) and borane tert-butylamine (0.05 g, 0.6 mmol) and continue to stir at room temperature for 2h to form a reaction solution, and then add methyl tert-butyl ether (50 mL) to the reaction solution, collect a precipitate by suction filtration, purify a residue with reverse-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain Compound 1 (white solid 0.28 g, yield 52%) after concentrating and drying.
(21) The calculated molecular weight of C.sub.87H.sub.100Cl.sub.2N.sub.10O.sub.27: 1786.61, the measured value: m/z=1787.60 [M+H].sup.+.
Example 2. Preparation of Compound 2
(22) Mix compound of general formula II (0.5 g, 0.3 mmol) with 10 mL DMF-methanol (1:1, v/v), add DIEA (0.1 mL, 0.6 mmol) and 4-phenylethoxybenzaldehyde (0.09 g, 0.4 mmol), stir at 65° C. for 2h and then cool to room temperature; add TFA (0.07 mL, 0.9 mmol) and borane tert-butylamine (0.05 g, 0.6 mmol) and continue to stir at room temperature for 2h to form a reaction solution, and then add methyl tert-butyl ether (50 mL) to the reaction solution, collect a precipitate by suction filtration, purify a residue with reversed-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain concentrated and dried to obtain compound 2 (white solid 0.31 g), yield 57%) after concentrating and drying.
(23) The calculated molecular weight of C.sub.88H.sub.102Cl.sub.2N.sub.10O.sub.27: 1800.63, the measured value: m/z=1801.63 [M+H]+.
Example 3. Preparation of Compound 3
(24) Mix compound II (1.0 g, 0.6 mmol) with 15 mL DMF-methanol (1:1, v/v), add DIEA (0.2 mL, 1.2 mmol) and 4-phenylpropoxybenzaldehyde (0.2 g, 0.8 mmol), stir at 65° C. for 2h and then cooled to room temperature; add TFA (0.14 mL, 1.8 mmol) and borane tert-butylamine (0.1 g, 1.2 mmol) and continue to stir at room temperature for 2h to form a reaction solution, then add methyl tert-butyl ether (70 mL) to the reaction solution, collect a precipitate by suction filtration, purify a residue with reverse-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain Compound 3 (white solid 0.65 g, yield 60%) after concentrating and drying.
(25) The calculated molecular weight of C.sub.89H.sub.104Cl.sub.2N.sub.10O.sub.27: 1814.64, the measured value: m/z=1815.64 [M+H]+.
Example 4. Preparation of Compound 4
(26) The preparation method of Compound 4 is the same as that of Compound 1, and the aldehyde used is replaced by 4-(4′-methylphenylmethoxy)benzaldehyde. Compound 4 (white solid 0.3 g, yield 56%) is obtained.
(27) The calculated molecular weight of C.sub.68H.sub.102Cl.sub.2N.sub.10O.sub.27: 1800.63, the measured value: m/z=1801.63 [M+H]+.
Example 5. Preparation of Compound 8
(28) Mix compound II (1.0 g, 0.6 mmol) with 15 mL DMF-methanol (1:1, v/v), add DMA (0.2 mL, 1.2 mmol) and 4-(4′-chlorophenylethoxy) benzaldehyde (0.21 g, 0.8 mmol), stir at 65° C. for 2 h, then cooled to room temperature; and then add TFA (0.14 mL, 1.8 mmol) and borane tert-butylamine (0.1 g, 1.2 mmol) and continue to stir at room temperature for 2h to form a reaction solution, then add methyl tert-butyl ether (70 mL) to the reaction solution, collect a precipitate by suction filtration, purify a residue with reverse-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain Compound 8 (white solid 0.55 g, yield 50%) after concentrating and drying.
(29) The calculated molecular weight of C.sub.88H.sub.101Cl.sub.3N.sub.10O.sub.27: 1834.59, the measured value: m/z=1835.60 [M+H]+.
Example 6. Preparation of Compound 10
(30) Mix Compound II (1.0 g, 0.6 mmol) with 15 mL DMF-methanol (1:1, v/v), add DIEA (0.2 mL, 1.2 mmol) and 4-(3′-bromophenylethoxy)benzaldehyde (0.24 g, 0.8 mmol), stir at 65° C. for 2h and then cool to room temperature; and then add TFA (0.14 mL, 1.8 mmol) and borane tert-butylamine (0.1 g, 1.2 mmol) and stir at room temperature for 2h to form a reaction solution, then add methyl tert-butyl ether (70 mL) to the reaction solution, collect a precipitate by suction filtration, purify a residue with reverse-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain Compound 10 (white solid 0.65 g, yield 58%) after concentrating and drying.
(31) The calculated molecular weight of C.sub.88H.sub.101BrCl.sub.2N.sub.10O.sub.27: 1878.54, the measured value: m/z=1879.54 [M+H]+.
Example 7. Preparation of Compound 11
(32) The preparation method of Compound 11 is the same as that of Compound 8, and the aldehyde used is replaced by 4-(4′-trifluoromethylphenylethoxy)benzaldehyde. Compound 11 (white solid 0.68 g, yield 61%) is obtained.
(33) The calculated molecular weight of C.sub.89H.sub.101Cl.sub.2F.sub.3N.sub.10O.sub.27: 1868.62, the measured value: m/z=1869.64 [M+H]+.
Example 8. Preparation of Compound 12
(34) Mix compound II (0.5 g, 0.3 mmol) with 10 mL DMF-methanol (1:1, v/v), add DIEA (0.1 mL, 0.6 mmol) and 4-(4′-methoxyphenylethoxy)benzaldehyde (0.1 g, 0.4 mmol), stir at 65° C. for 2h and then cool to room temperature; and then add TFA (0.07 mL, 0.9 mmol) and borane tert-butylamine (0.05 g, 0.6 mmol) and continue to stir at room temperature for 2h to form a reaction solution, and then add tert-butyl ether (50 mL) to the reaction solution, and collect a precipitate by suction filtration, purify a residue with reversed-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain Compound 12 (white solid 0.22 g, yield 40%) after concentrating and drying.
(35) The calculated molecular weight of C.sub.89H.sub.104Cl.sub.2N.sub.10O.sub.28: 1830.64, the measured value: m/z=1831.64 [M+H]+.
Example 9. Preparation of Compound 13
(36) The preparation method of Compound 13 is the same as that of Compound 12, and the aldehyde used is replaced by 4-(4′-chlorophenylpropoxy)benzaldehyde, to obtain Compound 13 (white solid 0.31 g, yield 56%).
(37) The calculated molecular weight of C.sub.89H.sub.103Cl.sub.3N.sub.10O.sub.27 1848.61, the measured value: m/z=1849.62 [M+H]+.
Example 10. Preparation of Compound 16
(38) Mix compound II (1.0 g, 0.6 mmol) with 15 mL DMF-methanol (1:1, v/v), add DIEA (0.2 mL, 1.2 mmol) and 4-(4′-chlorophenylethylthio) benzaldehyde (0.22 g, 0.8 mmol), stir at 65° C. for 2h and then cool to room temperature; and then add TFA (0.14 mL, 1.8 mmol) and borane tert-butylamine (0.1 g, 1.2 mmol) and continue to stir at room temperature for 2h to form a reaction solution, and then add methyl tert-butyl ether (70 mL) to the reaction solution, collect a precipitate by suction filtration, purify a residue with reverse-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain compound 16 (white solid 0.55 g, yield 50%) after concentrating and drying.
(39) The calculated molecular weight of C.sub.88H.sub.101Cl.sub.3N.sub.10O.sub.26S: 1850.57, the measured value: m/z=1851.57 [M+H]+.
Example 11. Preparation of Compound 18
(40) Stir Compound II (0.5 g, 0.3 mmol) with 10 mL DMF-methanol (1:1, v/v), add DIEA (0.1 mL, 0.6 mmol) and 4-(4′-methyl-N-Fmoc-benzylamino) benzaldehyde (0.18 g, 0.4 mmol), stir at 65° C. for 2h and then cool to room temperature, and then add TFA (0.07 mL, 0.9 mmol) and borane tert-butylamine (0.05 g, 0.6 mmol) and continue to stir at room temperature for 2h; and then add diethylamine (1 mL) for 3h under stirring again to form a reaction solution, add methyl tert-butyl ether (50 mL) to the reaction solution, collect a precipitate by suction filtration, and purify a residue with reverse-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain Compound 18 (white solid 0.24 g, yield 44%) after concentrating and drying.
(41) The calculated molecular weight of C.sub.88H.sub.103Cl.sub.2N.sub.11O.sub.26: 1799.65, the measured value: m/z=1800.65 [M+H]+.
Example 12. Preparation of Compound 20
(42) The preparation method of Compound 20 is the same as that of Compound 18, and the aldehyde used is replaced by 4-(N-Fmoc-phenylethylamino)benzaldehyde. Compound 20 (white solid 0.35 g, yield 65%) is obtained.
(43) The calculated molecular weight of C.sub.88H.sub.103Cl.sub.2N.sub.11O.sub.26: 1799.65, the measured value: m/z=1800.65 [M+H]+.
Example 13. Preparation of Compound 21
(44) Mix compound II (0.5 g, 0.3 mmol) with 10 mL DMF-methanol (1:1, v/v), add DIEA (0.1 mL, 0.6 mmol) and 4-(4′-chloro-N-Fmoc-phenylethylamino)benzaldehyde (0.19 g, 0.4 mmol), stir at 65° C. for 2h and then cool to room temperature, and then add TFA (0.07 mL, 0.9 mmol) and borane tert-butylamine (0.05 g, 0.6 mmol) and stir at room temperature for 2h, and then add diethylamine (1 mL) for 3h under stirring again to form a reaction solution, add methyl tert-butyl ether (50 mL) to the reaction solution, and then collect a precipitate by suction filtration; purify the residue with a reversed-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4, v/v), to obtain compound 21 (white solid 0.2 g, yield 36%) after concentrating and drying.
(45) The calculated molecular weight of C.sub.88H.sub.102Cl.sub.3N.sub.11O.sub.26: 1833.61, the measured value: m/z=1834.60 [M+H]+.
Example 14. Preparation of Compound 23
(46) The preparation method of Compound 23 is the same as that of Compound 21, and the aldehyde used is replaced by 4-(4′-methoxy-N-Fmoc-phenylethylamino)benzaldehyde. Compound 23 (white solid 0.3 g, yield 55%) is obtained.
(47) The calculated molecular weight of C.sub.89H.sub.105Cl.sub.2N.sub.11O.sub.27: 1829.66, the measured value: m/z=1830.65 [M+H]+.
Example 15. Preparation of Compound 25
(48) Mix compound of general formula II (0.5 g, 0.3 mmol) with 10 mL DMF-methanol (1:1, v/v), add DMA (0.1 mL, 0.6 mmol) and 4-(4′-chloro-N-Fmoc-amphetamine)benzaldehyde (0.2 g, 0.4 mmol), stir at 65° C. for 2h and then cool to room temperature, and then add TFA (0.07 mL, 0.9 mmol) and borane tert-butylamine (0.05 g, 0.6 mmol) and continue to stir at room temperature for 2h, and then add diethylamine (1 mL) and stir for 3h again to form a reaction solution, add methyl tert-butyl ether (50 mL) to the reaction solution, and then collect a precipitate by suction filtration; purify a residue with reversed-phase polymer filler, elute with methanol-0.04% TFA aqueous solution (1:4. V/v), to obtain compound 25 (white solid 0.36 g, yield 65%) after concentrating and drying.
(49) The calculated molecular weight of C.sub.89H.sub.104Cl.sub.3N.sub.11O.sub.26: 1847.62, the measured value: m/z=1848.63 [M+H]+.
Example 16. Example of Salt Formation
(50) Add 50 mg of Compound 8 to 1 mL of saturated hydrogen chloride methanol solution, stir at room temperature, and lyophilize it in order to obtain a white solid 50 mg of hydrochloride salt of Compound 8.
(51) In addition, respectively use hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, methanesulfonic acid, aspartic acid or glutamic acid instead of hydrogen chloride in the above-mentioned saturated hydrogen chloride methanol solution, in order to obtain a corresponding salt.
Example 17. Examples of Preparation
(52) It should be noted that the Example is only for illustration and is not intended to limit the scope of the present invention. The term “active ingredient” is referred to as compounds, solvates, tautomers, optical isomers, prodrugs, pharmaceutically acceptable salts, etc. of the present invention.
(53) A intravenous preparation can be prepared as follows:
(54) Active ingredient: 100 mg
(55) Isotonic saline: 1000 mL
(56) Solutions of the above components are intravenously administered to patients at a rate of 1 mL/min usually.
Example 18. Determination of the Antibacterial Activity of the Compound
(57) An antibacterial activity of the compounds in Table 1 is determined in vitro, and a minimum inhibitory concentration (MIC) is read. The determination method refers to the method of the Pharmacopoeia of the People's Republic of China (2015 edition). The MRSA detection strain is purchased from ATCC, and the VRE detection strain is from the clinically isolated drug-resistant strain 07-W3-45 from Shanghai Huashan Hospital. Known antibiotic oritavancin (phosphate) is used for a reference drug. The comparison test results are shown in the Table 2.
(58) A zebrafish toxicity test of the compounds in Table 1 is also tested. Randomly select wild AB zebrafish in a six-well plate, and inject each test sample at a dose of 50, 100, 150, 200, and 250 ng/tail intravenously. At the same time, a normal control group and a solvent control group (aqueous phosphoric acid) are set; During this experiment, the death of zebrafish is observed and recorded every day and the dead fish are removed. After 72 hours of treatment, the death of zebrafish is counted. Calculate the LD50 of each test product to zebrafish. The results are combined in Table 2.
(59) TABLE-US-00002 TABLE 2 MIC (μg/mL) zebrafish LD50 of each compound in Table 1 to MRSA and VRE MIC (μg/mL) ATCC43300 Clinical isolates ZebrafishLD.sub.50 Compound (MRSA) 07-W3-45 (VRE) (ng/tail) Orivancin 0.5 2 89.4 1 0.5 2 139 2 0.5 2 163 3 0.25 1 123 4 0.5 2 101 5 0.5 2 180 6 0.25 8 121 7 0.5 1 97 8 0.5 1 150 9 0.25 1 129 10 0.125 1 113 11 0.064 0.5 99 12 0.5 16 188 13 0.5 2 164 14 0.032 8 82 15 0.032 1 76 16 0.032 0.5 72 17 0.5 1 139 18 0.5 1 128 19 0.25 0.2 117 20 0.5 2 189 21 0.016 0.5 102 22 0.032 0.25 142 23 0.5 1 192 24 0.5 1 153 25 0.064 0.25 97
(60) All samples are tested for zebrafish toxicity in phosphate form.
(61) It can be seen from Table 2 that the glycopeptide compounds of the present invention have higher inhibitory activity or less toxicity against drug-resistant strains MRSA or VRE, and is higher security, compared to the second-generation glycopeptide drug oritavancin.
(62) The present invention is illustrated by the above examples, however, should understand that the present invention is not limited to special instance and implementation scheme described above. For those skilled in the art, any equivalent modifications and substitutions made to the invention are also within the scope of the invention. Therefore, all equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered within the scope of the present invention.