Quinolone compound as well as preparation method and anti-aging application thereof

Abstract

The invention relates to a quinolone compound as well as a preparation method and anti-aging application thereof. Specifically, the invention discloses a quinolone compound as shown in a formula I. The definitions of all groups and substituent groups in the quinolone compound are described in the specification. The invention also discloses pharmaceutically acceptable salts of the quinolone compound, a pharmaceutical composition of the quinolone compound, and medical application of the quinolone compound in the aspects of prolonging the service life and delaying and / or treating senescence. # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the fields of medicinal chemistry and pharmacotherapy, and in particular, to quinolone compounds, their preparation methods, and anti-aging uses. Background Art

[0002] Aging is a process in which the physiological functions of the body are impaired, the tissues and organs undergo degenerative changes, and ultimately lead to death as time goes by. Along with the aging of the body, the incidence of many chronic diseases such as heart disease, cancer, diabetes, Alzheimer's disease, etc. increases sharply, and they are more likely to occur in the later stage of life. At present, with the increase in the average human lifespan and the decline in the birth rate, the age structure of the global population is constantly changing. Now, the number of people over 65 years old is significantly higher than the number of people under 5 years old, and this trend will continue. Therefore, the degree of global aging is gradually increasing, which will significantly increase the operating costs of the national medical system. Therefore, delaying aging, preventing, delaying, alleviating, or even reversing the disease process related to age has become the key to solving the aging problem in countries around the world, and finding anti-aging drugs has become an urgent task in the medical field.

[0003] Therefore, there is an urgent need in this field to develop a compound with anti-aging or treating aging-related diseases. Summary of the Invention

[0004] The main object of the present invention is to provide a class of novel quinolone compounds, which are structurally modified based on enrofloxacin, have novel structures, and have good nematode lifespan extension activity.

[0005] In the first aspect of the present invention, there is provided a compound represented by Formula I, or a pharmaceutically acceptable salt thereof,

[0006]

[0007] Wherein,

[0008] R 1 is selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O, or S, substituted or unsubstituted benzyl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O, or S, substituted or unsubstituted C1-C6 alkyl-5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O, or S;

[0009] R 2Selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S;

[0010] R 6 Selected from the group consisting of: H, halogen, -OH, -NH2, -NO2, -CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl;

[0011] R 7 Selected from the group consisting of: -NR 10 、-OR 10 ;

[0012] Each R 10 is independently selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl; or two Rs 10 are linked to form a substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-2 heteroatoms selected from N, O or S;

[0013] Said substitution each independently means being substituted by 1-3 substituents selected from the group consisting of: -OH, -NH2, -NO2, -CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S.

[0014] In another preferred embodiment, R 1 is selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, substituted or unsubstituted benzyl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S, substituted or unsubstituted C1-C6 alkyl-5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S.

[0015] In another preferred embodiment, R 2 is selected from the group consisting of: substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S.

[0016] In another preferred embodiment, R 6 is selected from the group consisting of: H, halogen, -OH, -NH2, -NO2, -CN.

[0017] In another preferred example, R 6 is selected from the following group: H, halogen, -CH3, -CF3, -OCH3, -CN.

[0018] In another preferred example, each of the substitutions independently refers to being substituted by 1-3 substituents selected from the following group: halogen, -CH3, -CF3, -OCH3.

[0019] In another preferred example, the compound of formula I is the compound of formula II:

[0020]

[0021] wherein R 2 , R 6 , R 7 and R 10 are as defined above;

[0022] Each R 11 is independently selected from the following group: H, -OH, -NH2, -NO2, -CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, C6-C10 aryl, 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S.

[0023] In another preferred example, each R 11 is independently selected from the following group: H, -OH, -NH2, -NO2, -CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, C6-C10 aryl.

[0024] In another preferred example, R 2 is selected from the following group: substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S;

[0025] In another preferred example, R 6 is selected from the following group: H, halogen, -OH, -NH2, -NO2, -CN, substituted or unsubstituted C1-C6 alkyl.

[0026] In another preferred example, R 7 is NR 10 .

[0027] In another preferred example, each R 10Each independently selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl; or two R 10 are linked to form a substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-2 heteroatoms selected from N, O or S;

[0028] Said substitution each independently means being substituted by 1-3 substituents selected from the group consisting of: halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S.

[0029] In another preferred embodiment, the compounds are selected from the group consisting of:

[0030] 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-phenylquinolin-4(1H)-one; 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(thiophen-2-yl)quinolin-4(1H)-one; 7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 7-(4-ethylpiperazin-1-yl)-6-fluoro-1-(4-methoxybenzyl)-2-(4-methoxyphenyl)quinolin-4(1H)-one; 7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)-1-(4-(trifluoromethyl)benzyl)quinolin-4(1H)-one; 1-benzyl-7-(4-ethylpiperazin-1-yl)-2-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-6-carbonitrile; 1-benzyl-7-((3-chloropropyl)amino)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-6-fluoro-2-(4-methoxyphenyl)-7-(propylamino)quinolin-4(1H)-one; 1-benzyl-7-(cyclopropylmethoxy)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-6-fluoro-7-methoxy-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-6-fluoro-7-isopropoxy-2-(4-methoxyphenyl)quinolin-4(1H)-one.

[0031] In a second aspect of the present invention, there is provided a method for preparing a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, comprising the following steps:

[0032] In an inert solution, a compound of formula I c undergoes a nucleophilic substitution reaction with an H-R 7 compound to prepare a compound of formula I;

[0033]

[0034] wherein each group is as defined in the first aspect of the present invention.

[0035] The third aspect of the present invention provides a pharmaceutical composition comprising:

[0036] (i) a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof; and

[0037] (ii) a pharmaceutically acceptable carrier.

[0038] In another preferred embodiment, the pharmaceutical composition comprises:

[0039] (i) a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof; and

[0040] (ii) a second active ingredient having an anti-aging effect;

[0041] (iii) a pharmaceutically acceptable carrier.

[0042] In another preferred embodiment, the second active ingredient is selected from the group consisting of metformin, rapamycin, dasatinib, quercetin, curcumin, empagliflozin, canagliflozin, or a combination thereof.

[0043] The fourth aspect of the present invention provides the use of a compound as described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, for the preparation of a drug, the drug

[0044] (a) for extending lifespan;

[0045] (b) for retarding aging;

[0046] (c) for treating aging-related diseases.

[0047] In another preferred embodiment, the retarding of aging means being able to improve aging-related indicator factors, and the indicator factors are selected from the group consisting of cell cycle arrest-related proteins, DNA damage marker γ-H2AX, senescence-associated secretory phenotype factor SASP factor, senescence marker β-galactosidase SA-β-gal, and lamin B1.

[0048] In another preferred embodiment, the cell cycle arrest-related proteins are selected from the group consisting of p16, p21, and p53.

[0049] In another preferred embodiment, the senescence-associated secretory phenotype factor (SASP factor) is selected from the group consisting of: matrix metalloproteinase family (MMPs), chemokine CCL family and CXCLs family, interleukin IL family, and intercellular adhesion molecule ICAM.

[0050] In another preferred embodiment, the matrix metalloproteinase family (MMPs) is selected from the group consisting of: MMP-1, MMP-2 / MMP-3, MMP-7, MMP-9, MMP-13.

[0051] In another preferred embodiment, the matrix metalloproteinase family (MMPs) is selected from the group consisting of: MMP-1, MMP-3, MMP-7.

[0052] In another preferred embodiment, the chemokine CCL family and CXCLs family are selected from the group consisting of: CCL-2, CXCL-1, CXCL-3, CXCL-10.

[0053] In another preferred embodiment, the disease is selected from the group consisting of: tumor, acute or chronic renal failure, liver senescence, Alzheimer's disease, heart failure, neurodegenerative disease, hypertension, diabetes.

[0054] In a fifth aspect of the present invention, a method for delaying and / or treating senescence is provided, comprising the step of administering to a subject the compound described in the first aspect of the present invention, or a pharmaceutically acceptable salt thereof, or the composition described in the third aspect of the present invention.

[0055] In another preferred embodiment, the subject is a mammal, preferably a rodent or a primate mammal, more preferably a mouse, a rat or a human.

[0056] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as in the examples) can be combined with each other to form new or preferred technical solutions. Due to space limitations, they will not be repeated one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0057] Figure 1 It shows that the preferred compound I-02 has the activity of significantly improving the healthy lifespan of nematodes.

[0058] Figure 2 It shows that the preferred compound I-02 has the activity of significantly delaying cellular senescence at the animal level and delays cellular senescence as a Senomorphic drug.

[0059] Figure 3 It shows that the preferred compound I-02 has significant in vivo anti-aging activity at the animal level. DETAILED DESCRIPTION OF THE INVENTION

[0060] After extensive and in-depth research, the inventor of the present invention unexpectedly discovered a novel class of small molecule quinolone compounds that have lost antibacterial activity and simultaneously retain anti-aging activity through the optimization and modification of the lead structure of enrofloxacin. On this basis, the inventor completed the present invention.

[0061] Term

[0062] In the present invention, unless otherwise specified, the terms used have the general meanings known to those skilled in the art.

[0063] In the present invention, the halogen is F, Cl, Br or I.

[0064] In the present invention, the term "C1-C6 alkyl" refers to a straight-chain or branched-chain alkyl group having 1 to 6 carbon atoms, including but not limited to methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, tert-pentyl, and n-hexyl, etc. Similar terms such as "C1-C4 alkyl" have similar definitions.

[0065] In the present invention, the term "C1-C6 alkoxy" refers to a straight-chain or branched-chain alkoxy group having 1 to 6 carbon atoms, including but not limited to methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, etc. Similar terms such as "C1-C3 alkoxy" have similar definitions.

[0066] In the present invention, the term "C2-C6 alkenyl" refers to a straight-chain or branched-chain alkenyl group having 2 to 6 carbon atoms and containing at least one double bond, including but not limited to vinyl, propenyl, butenyl, isobutenyl, pentenyl, and hexenyl, etc. Similar terms such as "C2-C6 alkenyl" have similar definitions.

[0067] In the present invention, the term "C2-C6 alkynyl" refers to a straight-chain or branched-chain alkynyl group having 2 to 6 carbon atoms and containing at least one triple bond, including but not limited to ethynyl, propynyl, butynyl, isobutynyl, pentynyl, and hexynyl, etc. Similar terms such as "C2-C6 alkynyl" have similar definitions.

[0068] In the present invention, the term "C3-C8 cycloalkyl" represents a cyclic aliphatic hydrocarbon group composed of 3 to 8 ring-forming carbon atoms, and so on; it should be understood that the "cycloalkyl" described in the present invention includes not only monocyclic aliphatic hydrocarbon groups, but also fused-ring, spiro-ring, and bridged-ring systems composed of multiple cyclic aliphatic hydrocarbons; the "cycloalkyl" described in the present invention includes not only aliphatic hydrocarbon groups with completely saturated carbon atoms, but also aliphatic hydrocarbon groups with some unsaturated bonds in the carbon atoms; examples of the "cycloalkyl" described in the present invention include but not limited to: When "cycloalkyl" is used as a substituent, the connection site with the main body of the molecule can occur at any position on the "cycloalkyl" that allows chemical bonds. "C3-C6 cycloalkyl" and other similar terms have similar definitions.

[0069] In the present invention, the term "aryl" refers to a monocyclic system and a bicyclic system composed of a specific number of carbon atoms and complying with Hückel's rule; it should be understood that when the "aryl" described in the present invention is a bicyclic system, it not only includes the case where all rings are aromatic rings, but also includes the case where only one ring is an aromatic ring and the other ring is a non-aromatic aliphatic ring.

[0070] In the present invention, the term "C6-C10 aryl" refers to a ring system having 6 to 10 carbon atoms, at least one of which is an aromatic ring; examples of the "aryl" described in the present invention include but are not limited to Etc.; when "aryl" is used as a substituent, the connection site with the main body of the molecule occurs on the aromatic ring.

[0071] In the present invention, the term "heterocycloalkyl" refers to a cyclic group that indicates a specific number of ring atoms, contains at least one ring heteroatom (N, O or S), is saturated or partially unsaturated, and is non-aromatic; it should be understood that the "heterocyclic group" described in the present invention includes not only monocyclic heterocyclic ring systems, but also polycyclic heterocyclic ring systems, such as cyclic, spirocyclic and bridged rings; when the "heterocyclic group" is a polycyclic system, at least one ring contains a ring heteroatom, and the other rings may contain ring heteroatoms or may be cycloalkyl; for example, the term "4-8 membered heterocycloalkyl" refers to a monocyclic or polycyclic system with 4 to 8 ring atoms, at least one of which is a heteroatom, saturated or partially unsaturated; the definitions of other similar terms are similar; preferably, the number of heteroatoms is 1 to 3. Including (but not limited to) the following groups: etc.; it should be understood that when a "heterocyclic group" is used as a substituent, the connection site with the main body of the molecule can occur at any position on the "heterocyclic group" that is allowed by a chemical bond.

[0072] In the present invention, the term "heteroaryl" refers to a cyclic group having a specific number of ring-forming atoms, containing at least 1 ring-forming heteroatom (N, O or S), and having aromaticity; unless otherwise defined, the "heteroaryl" described in the present invention includes not only monocyclic heteroaryl systems, but also polycyclic heteroaryl systems, such as bicyclic heteroaryl, tricyclic heteroaryl, tetracyclic heteroaryl; when the "heteroaryl" is a polycyclic heteroaryl system, at least one of the rings is aromatic, and the other rings can be aromatic or non-aromatic, and the heteroatoms can be in the aromatic rings or in the non-aromatic rings; the polycyclic heteroaryl systems include not only fused-ring systems, but also bridged-ring and spiro-ring systems. The term "5-7 membered heteroaryl" refers to a cyclic group having 5 to 7 ring-forming atoms, at least one of which is a heteroatom and has aromaticity. The definitions of other similar terms can be deduced by analogy.

[0073] In the present invention, the term "halogenated" means being substituted by a halogen.

[0074] In the present invention, the term "optionally" means that when there is a series of candidate groups for selection, some of them can be selected, or none of them can be selected.

[0075] The "independently of each other" described in the present invention means that when several substituents defined simultaneously are selected from the same series of candidate groups, they do not affect each other, and they can be the same or different.

[0076] In the present invention, the term "substituted" means that one or more hydrogen atoms on a specific group are replaced by specific substituents. The specific substituents are the substituents described correspondingly in the foregoing text or the substituents appearing in each embodiment. Unless otherwise specified, a substituted group can have a substituent selected from a specific group at any substitutable site of the group, and the substituents can be the same or different at each position. Those skilled in the art should understand that the combinations of substituents expected by the present invention are those that are stable or chemically achievable.

[0077] In the present invention, the term "1-6" means having 1, 2, 3, 4, 5 or 6, and other similar terms independently have similar meanings.

[0078] It should be understood that when a certain group is present at multiple different positions in a compound, its definitions at each position are independent of each other and can be the same or different. That is to say, the term "selected from the group consisting of" has the same meaning as the term "independently selected from the group consisting of".

[0079] Compound

[0080] For the first or second aspect of the present invention, the compound represented by the general formula (I) may contain one or more chiral centers, and enantiomers and diastereomers exist. The compound represented by the general formula (I) of the present invention may also contain many geometric isomers such as alkenes, C=N double bonds, amides, etc. Unless otherwise specified, all the above-mentioned chiral (enantiomers, diastereomers, axial chiral isomers), racemates, cis geometric isomers, trans geometric isomers, mixtures of cis and trans geometric isomers, rotamers and their mixtures are included in the present invention. Those of ordinary skill in the art can use common separation or synthesis methods in the laboratory to separate or prepare the compounds containing asymmetric centers in the present invention to obtain single isomers. For example, for enantiomers, two enantiomers can be obtained by using general chiral resolution methods or asymmetric synthesis methods. For diastereomers, they can be separated by methods such as fractional recrystallization or chromatographic separation, which does not destroy the novelty of the compounds of the present invention.

[0081] The compounds of the present invention have the structure shown in Formula I:

[0082]

[0083] Wherein, the definitions of each group are as above.

[0084] The present invention also provides a compound having the structure shown in Formula II:

[0085]

[0086] Wherein, the definitions of each group are as above.

[0087] As used herein, the term "pharmaceutically acceptable salt" refers to a salt formed by a positively charged group on the compound represented by the general formula (I) and an anion, or a salt formed by a negatively charged group on the compound represented by the general formula (I) and a cation. Suitable anions include but are not limited to chloride ion, bromide ion, iodide ion, sulfate ion, nitrate ion, phosphate ion, citrate ion, methanesulfonate ion, trifluoroacetate ion, acetate ion, malate ion, tosylate ion, tartrate ion, fumarate ion, glutamate ion, glucuronate ion, lactate ion, glutarate ion or maleate ion, etc. Suitable cations include but are not limited to sodium ion, potassium ion, magnesium ion, calcium ion, ammonium ion, etc.

[0088] In another preferred embodiment, the pharmaceutically acceptable salts of the present invention refer to salts formed by the compounds represented by the general formula (I) and the following acids, such as but not limited to: hydrofluoric acid, hydrochloric acid, hydrobromic acid, phosphoric acid, acetic acid, oxalic acid, sulfuric acid, nitric acid, methanesulfonic acid, aminosulfonic acid, salicylic acid, trifluoromethanesulfonic acid, naphthalenesulfonic acid, maleic acid, citric acid, acetic acid, lactic acid, tartaric acid, succinic acid, oxalacetic acid, pyruvic acid, malic acid, glutamic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, ethanesulfonic acid, naphthalenedisulfonic acid, malonic acid, fumaric acid, propionic acid, oxalic acid, trifluoroacetic acid, stearic acid, pamoic acid, hydroxymaleic acid, phenylacetic acid, benzoic acid, glutamic acid, ascorbic acid, p-aminobenzenesulfonic acid, 2-acetoxybenzoic acid, and hydroxyethanesulfonic acid, etc.; or salts formed by the compounds represented by the general formula (I) and inorganic bases, such as but not limited to sodium salts, potassium salts, calcium salts, aluminum salts or ammonium salts; or salts formed by the compounds represented by the general formula (I) and organic bases, such as but not limited to methylamine salts, ethylamine salts, ethanolamine salts, ammonium salts of hydroxymethylaminomethane (TRIS), etc.

[0089] The compound of the general formula (I) of the present invention or its pharmaceutically acceptable salt is precipitated, crystallized or recrystallized from water or an organic solvent, and the compound may contain solvent molecules used. In addition, different crystallization conditions may result in different crystal forms of the compound. Therefore, the compounds of the general formula (I) containing different chemical dosages of crystallization solvents and all crystal forms, as well as their pharmaceutically acceptable salts, are within the scope of the present invention.

[0090] Preparation method

[0091] The embodiments of the present invention specifically describe the preparation methods of the compounds of the structure of formula (I) of the present invention, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention can also be conveniently prepared by optionally combining various synthesis methods described in this specification or known in the art, and such combinations can be easily carried out by those skilled in the art to which the present invention pertains.

[0092] Typically, the preparation process flow of the compounds of the present invention is as follows. Unless otherwise specified, the raw materials and reagents used can be purchased through commercial channels.

[0093] The preparation method of the compound represented by formula I of the present invention is as follows:

[0094] Route 1:

[0095]

[0096] Step 1: In a dry two-necked round-bottom flask, under nitrogen protection, dissolve aromatic acetylene with different R2 groups (1.1 equivalents, 2.2 mmol) in 5 mL of anhydrous tetrahydrofuran, then slowly add n-butyllithium (1 equivalent, 2 mmol), and stir at -78 °C for 30 minutes. Dissolve the benzaldehyde derivative (1 equivalent, 2 mmol) in anhydrous tetrahydrofuran solution and drop it into the reaction, and continue the reaction at -78 °C for 4 - 5 hours. Until the mixture reacts completely, transfer the flask to room temperature and stir for 30 minutes, and carefully quench n-butyllithium with ice water. Extract the reaction solution with EA and collect the organic layer. After drying with anhydrous sodium sulfate, purify by column chromatography (SiO2, PE:EA = 10:1) to obtain the pure intermediate I a 。

[0097]

[0098] Step 2: Dissolve intermediate I a (1 equivalent) in 5 mL of ultradry dichloromethane solvent, then slowly add 1.2 equivalents of Dess-Martin reagent under ice bath conditions. After adding, continue to stir the reaction system at room temperature for 2 hours. After the reaction is completed, cool it to room temperature and extract it three times with EA and saturated ammonium chloride aqueous solution. Collect the organic phase, dry it with anhydrous sodium sulfate, and purify by column chromatography (SiO2, PE:EA = 20:1) to obtain intermediate I b 。

[0099]

[0100] Step 3: Dissolve intermediate I b (1 equivalent), potassium phosphate (1.5 equivalents) and R 1 -NH2 (3 equivalents) in 5 mL of dimethyl sulfoxide and stir at 140 °C for 2 hours. After the reaction is completed, cool it to room temperature and extract it three times with EA and saturated ammonium chloride aqueous solution. Collect the organic phase, dry it with anhydrous sodium sulfate, and purify by column chromatography (SiO2, PE:EA = 10:1) to obtain intermediate I c 。

[0101]

[0102] Step 4: In a dry round-bottom flask, dissolve the above intermediate I c (1 equivalent) in 2 mL of dimethyl sulfoxide. Add H-R 7(2 equivalents), stirred at 70°C for 24-48 hours. After the reaction, extracted with EA and brine 3-5 times to remove the DMSO solvent. After drying with anhydrous sodium sulfate, purified by column chromatography (SiO2, DCM:MeOH=10:1) to obtain quinolone compounds I-01, I-02 and I-03.

[0103] Route 2:

[0104]

[0105] Step 1: While stirring, iodine (1.2 equivalents, 60 mmol) was slowly added to a solution of the differently substituted aniline starting material (1 equivalent, 50 mmol) in 50 ml of deionized water. The reaction was stirred continuously at room temperature for 2 hours. After completion, the reaction was stopped and extracted with aqueous sodium thiosulfate solution and EA. Subsequently, the organic phase was dried over anhydrous sodium sulfate. Intermediate I was purified by column chromatography (SiO2, PE:EA=20:1). d .

[0106]

[0107] Step 2: In a dry two-necked round-bottom flask, place intermediate I d (1 equivalent), molybdenum hexacarbonyl (1.5 equivalents), palladium acetate (0.02 equivalents) and tri-tert-butylphosphine tetrafluoroborate (0.06 equivalents) were dissolved in 15 ml of ultra-dry acetonitrile. The flask was sealed, the air was replaced with nitrogen, and stirred at room temperature. Triethylamine (2 equivalents) and aromatic acetylene with different R2 groups (1.2 equivalents) were slowly added dropwise with a syringe. After stirring for 16 hours, the mixture was extracted three times with saturated aqueous ammonium chloride solution and EA, dried over anhydrous sodium sulfate, and purified by column chromatography (SiO2, PE:EA=8:1) to obtain pure intermediate I e .

[0108]

[0109] Step 3: Intermediate I e Dissolve in 15 ml of diethylamine and stir overnight at room temperature. Wash the reactant with water and extract with EA three times. Purify by column chromatography (SiO2, DCM:MeOH=5:1) to obtain intermediate I f .

[0110]

[0111] Step 4: In a dry round-bottom flask, place intermediate I f(1 equivalent) was dissolved in 3 mL of ultradry N,N-dimethylformamide and stirred in an ice bath. After sodium hydride (1.2 equivalents) was slowly added to the mixture and stirred for 30 minutes, the iodoalkane or bromoalkane (1.2 equivalents) with different R1 groups was dropped into it. After addition, the reaction flask was removed from the ice bath. After 2 hours, the reaction was monitored by TLC to determine whether to heat under reflux. After the reaction was completed, it was extracted 3 - 5 times with EA and brine to remove the solvent. Then the organic layer was concentrated under vacuum and purified by column chromatography (SiO2, PE:EA = 10:1 / SiO2, DCM:MeOH = 40:1) to obtain the pure intermediate I g .

[0112]

[0113] Step Five: In a dry two-necked round-bottom flask, intermediate I g (1 equivalent), tris(dibenzylideneacetone)dipalladium(0) (0.05 equivalent), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.05 equivalent) and sodium tert-butoxide (2 equivalents) were added and the reaction system was sealed. Then 3 mL of toluene was added as the solvent. The air in the reaction system was displaced with nitrogen, and the R 10 -NH2 or (R 10 )2-NH compound raw material (1.2 equivalents) was dropped into the reaction system and sealed. It was stirred at 120 °C for 4 - 5 hours. After the reaction was completed, it was cooled to room temperature, and the catalyst was removed by suction filtration with diatomaceous earth. The filtrate was evaporated under vacuum to remove the solvent. Finally, it was purified by column chromatography (SiO2, DCM:MeOH = 10:1 / DCM:MeOH = 5:1) to obtain quinolone compounds I-04 to I-10

[0114] Route Three:

[0115] Steps One to Four are the same as Route Two

[0116]

[0117] Step Five: In a dry two-necked round-bottom flask, intermediate I g(1 equivalent), 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl(2-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (0.1 equivalent) and cesium carbonate (2 equivalents) were added and the reaction system was sealed. Then, 3 mL of toluene was added as a solvent. The air in the reaction system was displaced with nitrogen, and 1.2 equivalents of trifluoroethanol was added dropwise to the reaction system with a syringe and sealed. Subsequently, the mixture was stirred at 90 °C for 4 - 5 h. After the reaction was completed, it was cooled to room temperature, and the catalyst was removed by suction filtration through diatomaceous earth. The filtrate was evaporated under vacuum to remove the solvent. Finally, the intermediate I was purified by column chromatography (SiO2, PE:EA = 10:1). h 。

[0118]

[0119] Step VI: In a dry round-bottom flask, the intermediate I h (1 equivalent) was dissolved in 3 mL of anhydrous N,N-dimethylformamide, and then different iodoalkane raw materials R 10 I (1.2 equivalents) and 2 equivalents of cesium carbonate were added, and the mixture was stirred at room temperature or with heating. After the reaction was completed, it was extracted 3 - 5 times with EA and brine to remove the solvent. Then, the organic layer was concentrated under vacuum and purified by column chromatography (SiO2, PE:EA = 10:1 / DCM:MeOH = 40:1) to obtain quinolone compounds I-11 to I-13.

[0120] Among them, R 1 、R 2 、R 6 、R 7 and R 10 are as defined above.

[0121] Drug Compositions and Administration Methods

[0122] The pharmaceutical composition of the present invention comprises a compound of the present invention or a pharmaceutically acceptable salt thereof within a safe and effective amount range, and a pharmaceutically acceptable excipient or carrier. Wherein the "safe and effective amount" means that, compared with an object not receiving the treatment of this dose, the object receiving the treatment of this dose has the lesion or side effect, etc. cured, improved, effectively prevented, or the incidence rate thereof significantly reduced; in addition, it also includes an effective dose for enhancing normal physiological functions. "Safe" means that the amount of the active ingredient is sufficient to significantly improve the condition without causing serious side effects.

[0123] In the pharmaceutical composition, the compound of the present invention is used as an active ingredient, and its weight accounts for 0.1-99.9% of the total weight of the pharmaceutical composition, and the rest are pharmaceutical excipients; the preferred ratio of the compound of the present invention to the excipients is: the compound of the present invention as an active ingredient accounts for more than 60% of the total weight, and the rest accounts for 0-40% of the total weight, and the amount of the rest is preferably 1-20%, and most preferably 1-10%. Usually, the pharmaceutical composition contains 1-2000 mg of active ingredient per dose, and more preferably, it contains 10-200 mg of active ingredient per dose. Preferably, the "per dose" is a tablet.

[0124] The "pharmaceutical excipients" are pharmaceutically acceptable carriers, excipients, sustained-release agents, odorants, flavorants, etc. The "pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances, which are suitable for human use and must have sufficient purity and sufficiently low toxicity. "Compatibility" here means that the components in the composition can be mixed with the active ingredient of the present invention and with each other without significantly reducing the efficacy of the active ingredient. Examples of pharmaceutically acceptable carriers include but are not limited to cellulose and its derivatives (such as sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricants (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oils (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyols (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifiers (such as ) and wetting agents (such as sodium dodecyl sulfate), etc.

[0125] In addition to containing the compound shown in the general formula (I) as an active ingredient, the pharmaceutical composition may further contain one or more other therapeutic agents. Among them, the "other therapeutic agent" is a therapeutic agent for central nervous system diseases.

[0126] Adjuvants commonly used in the preparation of pharmaceutical compositions may also be included. The "adjuvant" is a flavoring agent, pigment, preservative and antioxidant, such as vitamin E, vitamin C, BHT and BHA.

[0127] The compound or pharmaceutical composition of the present invention can be made into various dosage forms based on the conventional processes in the field of pharmaceutical preparations, such as tablets, capsules, powders, syrups, solutions, suspensions, sprays, creams, ointments, gels, transdermal patches, microneedles, etc., and can exist in a suitable solid or liquid carrier or diluent. The pharmaceutical composition of the present invention can also be stored in a suitable sterilized apparatus for injection or infusion. From the standpoint of easy preparation and administration, the preferred pharmaceutical composition is a solid composition, especially tablets and solid-filled or liquid-filled capsules.

[0128] The compounds or pharmaceutical compositions of the present invention can be clinically used in mammals, including humans and animals. There is no particular limitation on the administration mode, and representative administration modes include but are not limited to oral administration, nasal inhalation, topical skin administration, intravenous injection, intramuscular injection, subcutaneous injection, etc. Preferably, the preferred administration route of the compounds or pharmaceutical compositions of the present invention is oral administration.

[0129] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or calcium phosphate, or mixed with the following components: (a) fillers or bulking agents, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, such as hydroxypropylmethyl cellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, such as glycerol; (d) disintegrants, such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solubilizing agents, such as paraffin wax; (f) absorption accelerators, such as quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, such as kaolin; and (i) lubricants, such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain buffering agents.

[0130] Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and casings, such as enteric coatings and other materials well known in the art. They may contain opacifying agents, and the release of the active compound or compounds in such compositions can be delayed in a portion of the digestive tract. Examples of embedding components that can be used are polymeric substances and waxes. If necessary, the active compound can also be in the form of microcapsules with one or more of the above excipients.

[0131] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active ingredient, the liquid dosage forms may contain inert diluents conventionally used in the art, such as water or other solvents, solubilizing agents, and emulsifying agents, such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3 - butanediol, dimethylformamide, and oils, especially cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances.

[0132] In addition to these inert diluents, the compositions may also contain adjuvants, such as wetting agents, emulsifying agents, and suspending agents, sweetening agents, flavoring agents, and fragrances.

[0133] In addition to the active ingredient, the suspension may contain suspending agents, such as ethoxylated isooctadecanol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide, agar, or a mixture of these substances, etc.

[0134] Injectable preparations include, but are not limited to, sterile, injectable, aqueous, oily solutions, suspensions, emulsions, etc. These preparations can also be formulated with suitable parenteral diluents, dispersants, wetting agents, suspending agents, etc. Such injectable preparations can be sterilized by filtration through a bacteria-retaining filter. These preparations can also be formulated with bactericides, which are dissolved or dispersed in the injectable medium or by other methods known in the art.

[0135] Dosage forms of the compounds of the present invention for topical administration include ointments, powders, patches, sprays and inhalants. The active ingredient is mixed with a physiologically acceptable carrier and any preservatives, buffers, or propellants that may be necessary under sterile conditions.

[0136] The treatment method of the present invention can be administered alone, or in combination with other treatment means or therapeutic drugs (such as antidepressant drugs).

[0137] When using the pharmaceutical composition, a safe and effective amount of the compound of the present invention is applied to a mammal in need of treatment (such as a human), wherein the dosage during administration is an effective dosage considered pharmaceutically. For a person weighing 60 kg, the daily dosage is usually 1 - 2000 mg, preferably 50 - 1000 mg. Of course, the specific dosage should also consider factors such as the administration route and the patient's health condition, which are within the scope of the skills of a skilled physician.

[0138] When used as a pharmaceutical preparation, the compounds or pharmaceutical compositions shown in the present invention can be used once a day or in divided doses. Regardless of the method of use, the optimal dosage for an individual should be determined according to the specific treatment. Usually, it starts with a small dose and gradually increases the dose until the most suitable dose is found.

[0139] Compared with the prior art, the present invention has the following main advantages:

[0140] 1. The present invention discovers for the first time that the said compound has excellent anti-aging effects.

[0141] 2. The compound of the present invention has an anti-aging effect that does not depend on antibacterial effects.

[0142] 3. The compound of the present invention can regulate the expression levels of biomarkers related to aging, thereby effectively improving aging-related indicators.

[0143] The present invention will be further described below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and not to limit the scope of the present invention. The experimental methods without specific conditions in the following embodiments are generally carried out under conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989), or according to the conditions recommended by the manufacturer. Unless otherwise specified, percentages and parts are weight percentages and weight parts.

[0144] Preparation of Compounds

[0145] Example 1

[0146] Preparation of 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-phenylquinolin-4(1H)-one (I-01):

[0147]

[0148] In a dry two-necked round-bottom flask, under nitrogen protection, phenylacetylene (1.1 equivalents, 2.2 mmol) was dissolved in 5 mL of anhydrous tetrahydrofuran, and then n-butyllithium (1 equivalent, 2 mmol) was slowly added, and the mixture was stirred at -78 °C for 30 minutes. A solution of 2 mL of 2-bromo-4,5-difluorobenzaldehyde (442 mg, 2 mmol) in anhydrous tetrahydrofuran was added dropwise to the reaction, and the reaction was continued at -78 °C for 4 - 5 hours. After the mixture reacted completely, the flask was transferred to room temperature and stirred for 30 minutes, and n-butyllithium was carefully quenched with ice water. The solution was extracted with EA and the organic layer was collected. After drying over anhydrous sodium sulfate, it was purified by column chromatography (SiO2, PE:EA = 10:1) to obtain the pure intermediate 1-(2-bromo-4,5-difluorophenyl)-3-phenylprop-2-yn-1-ol (I a -01). White solid, 516.96 mg, yield 80%.

[0149] The above intermediate 1-(2-bromo-4,5-difluorophenyl)-3-phenylprop-2-yn-1-ol (I a -01) (516.96 mg) was dissolved in 5 mL of ultradry dichloromethane solvent, and then 1.2 equivalents of Dess-Martin reagent was slowly added under ice bath conditions. After addition, the reaction system was stirred at room temperature for 2 hours. After the reaction was completed, it was cooled to room temperature and extracted three times with EA and saturated aqueous ammonium chloride solution. The organic phase was collected, dried over anhydrous sodium sulfate, and purified by column chromatography (SiO2, PE:EA = 20:1) to obtain the intermediate 1-(2-bromo-4,5-difluorophenyl)-3-phenylprop-2-yn-1-one (I b-01). White solid, 421.27 mg, yield 82%. 1 1H NMR (400 MHz, DMSO-d6) δ 8.23 (dd, J = 10.7, 8.3 Hz, 1H), 8.08 (dd, J = 10.1, 7.2 Hz, 1H), 7.82–7.75 (m, 2H), 7.69–7.59 (m, 1H), 7.54 (t, J = 7.5 Hz, 2H).

[0150] Dissolve the above intermediate 1-(2-bromo-4,5-difluorophenyl)-3-phenylprop-2-yn-1-one (I b -01) (421.27 mg), potassium phosphate (1.5 equiv) and cyclopropylamine (3 equiv) in 5 mL of dimethyl sulfoxide and stir at 140 °C for 2 h. After completion of the reaction, cool it to room temperature and extract three times with EA and saturated aqueous ammonium chloride. Collect the organic phase, dry it over anhydrous sodium sulfate and purify it by column chromatography (SiO2, PE:EA = 10:1) to obtain the intermediate 1-cyclopropyl-6,7-difluoro-2-phenylquinolin-4(1H)-one (I c -01). White solid, 187.21 mg, yield 48%. 1 1H NMR (400 MHz, Acetone-d6) δ 8.13–7.90 (m, 2H), 7.74–7.66 (m, 2H), 7.61–7.50 (m, 3H), 6.04 (s, 1H), 3.61 (tt, J = 6.9, 4.0 Hz, 1H), 1.01 (d, J = 7.1 Hz, 2H), 0.63 (dd, J = 4.1, 1.5 Hz, 2H).

[0151] In a dry round-bottom flask, dissolve the above intermediate 1-cyclopropyl-6,7-difluoro-2-phenylquinolin-4(1H)-one (I c -01) (187.21 mg) in 2 mL of dimethyl sulfoxide. Add N-ethylpiperazine (2 equiv) to the reaction and stir at 70 °C for 48 h. After completion of the reaction, extract 3–5 times with EA and brine to remove the DMSO solvent. After drying over anhydrous sodium sulfate, purify it by column chromatography (SiO2, DCM:MeOH = 10:1) to obtain the quinolone compound 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-phenylquinolin-4(1H)-one I-01. White solid, 164.18 mg, yield 66.6%. 11H NMR(400MHz,DMSO-d6)δ7.79–7.33(m,7H),5.94(s,1H),3.57(s,1H),3.24(t,J=4.9Hz,4H),2.59(t,J=4.7Hz,4H),2.42(d,J=7.2Hz,2H),1.35(d,J=5.7Hz,1H),1.05(t,J=7.1Hz,3H),0.88(dd,J=13.8,6.8Hz,3H); HRMS: calcd for C 24 H 26 FN3O[M+H] + , 392.2133; found 392.2118; 13 13C NMR(101MHz,DMSO-d6)δ175.40, 155.33, 151.14, 143.89, 143.78(d), 140.97, 137.14, 129.50, 128.84, 128.81, 128.74, 120.96, 120.90(d), 111.81, 111.68(d), 110.47, 108.17, 52.60, 52.60, 52.01, 50.23, 50.23, 32.75, 32.65(d), 12.86, 12.40, 12.40。

[0152] Example 2

[0153] Preparation of 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-02):

[0154]

[0155] Except that the reaction raw material phenylacetylene in (1) of Example 1 was replaced with 4-ethynylanisole, the rest of the required raw materials, reagents and preparation methods were the same as those in Example 1, and the total reaction yield was 73%. This compound is a white solid, 11H NMR(400 MHz, DMSO-d6) δ 7.69 (d, J = 13.3 Hz, 1H), 7.62–7.54 (m, 2H), 7.45 (d, J = 7.5 Hz, 1H), 7.11–7.01 (m, 2H), 5.92 (s, 1H), 3.84 (s, 3H), 3.58 (tt, J = 6.9, 4.0 Hz, 1H), 3.22 (d, J = 4.9 Hz, 4H), 2.58 (s, 4H), 2.41 (q, J = 7.1 Hz, 2H), 1.34 (d, J = 5.9 Hz, 1H), 1.23 (d, J = 3.7 Hz, 2H), 1.05 (t, J = 7.1 Hz, 3H), 0.93 (d, J = 7.3 Hz, 1H); HRMS: calcd for C 25 H 28 FN3O2 [M+H] + , 422.2238; found 422.2233; 13 13C NMR(101 MHz, DMSO-d6) δ 175.36, 160.20, 155.22, 151.08, 143.76, 140.99, 130.32, 130.32, 129.37, 114.21, 114.21, 111.43, 110.68, 110.46 (d), 55.70, 52.57, 52.57, 52.00, 50.18, 50.18, 32.65, 12.99, 12.37, 12.37。

[0156] Example 3

[0157] Preparation of 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(thiophen-2-yl)quinolin-4(1H)-one (I-03):

[0158]

[0159] Except that the reaction raw material phenylacetylene in (1) of Example 1 was replaced with 2-ethynylthiophene, the rest of the required raw materials, reagents and preparation methods were the same as in Example 1, and the total reaction yield was 71.3%. This compound is a white solid, 11H NMR (400 MHz, DMSO-d6) δ 7.85–7.82 (m, 1H), 7.67 (d, J = 13.3 Hz, 1H), 7.58 (d, J = 1.2 Hz, 1H), 7.45 (d, J = 7.5 Hz, 1H), 7.25–7.22 (m, 1H), 6.10 (s, 1H), 3.67 (dq, J = 6.8, 3.5 Hz, 1H), 3.25 (d, J = 4.9 Hz, 4H), 2.69–2.53 (m, 4H), 1.23 (d, J = 3.6 Hz, 2H), 1.06 (t, J = 7.1 Hz, 5H), 0.59–0.54 (m, 2H); HRMS: calcd for C 22 H 24 FN3OS [M+H] + , 398.1697; found 398.1684; 13 13C NMR (101 MHz, DMSO-d6) δ 175.27, 151.19, 148.27, 143.99, 143.88 (d), 141.10, 137.20, 130.37, 129.43, 128.31, 121.05, 112.05, 111.86 (d), 110.63, 108.25, 52.60, 52.60, 52.01, 50.22, 50.22, 32.98, 13.05, 12.41, 12.41。

[0160] Example 4

[0161] Preparation of 7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-04):

[0162]

[0163] (1) While stirring, iodine (1.2 eq, 60 mmol) was slowly added to a solution of 3-bromo-4-fluoroaniline (1 eq, 9.5 g) in 50 mL of deionized water. The reaction was continuously stirred at room temperature for 2 h. After completion, the reaction was stopped and extracted with aqueous sodium thiosulfate and EA. Subsequently, the organic phase was dried over anhydrous sodium sulfate. Purification by column chromatography (SiO2, PE:EA = 20:1) gave the intermediate 5-bromo-4-fluoro-2-iodoaniline (I d -04). White solid, 8.8 g, yield 54%. 1 1H NMR (400 MHz, CDCl3) δ 7.42 (d, J = 7.6 Hz, 1H), 6.93 (d, J = 6.0 Hz, 1H), 4.01 (s, 2H).

[0164] (2) In a dry two-necked round-bottom flask, the intermediate (I d -04) (8.8 g), molybdenum hexacarbonyl (1.5 eq.), palladium acetate (0.02 eq.) and tri-tert-butylphosphine tetrafluoroborate (0.06 eq.) were dissolved in 15 ml of ultra-dry acetonitrile. The flask was sealed, the air was replaced with nitrogen, and stirred at room temperature. Triethylamine (2 eq.) and 4-ethynylanisole (1.2 eq.) were slowly added dropwise with a syringe. After stirring at room temperature for 16 hours, the mixture was extracted three times with saturated aqueous ammonium chloride solution and EA, dried over anhydrous sodium sulfate, and purified by column chromatography (SiO2, PE:EA=8:1) to obtain the pure intermediate 1-(2-amino-4-bromo-5-fluorophenyl)-3-(4-methoxyphenyl)prop-2-yn-1-one (I e -04). Yellow solid, 4.5 g, yield 46.3%. 1 H NMR (400MHz, CDCl3) δ7.91 (d, J = 9.2Hz, 1H), 7.68–7.62 (m, 2H), 6.99–6.90 (m, 3H), 6.26 (s, 2H), 3.89 (s, 3H).

[0165] (3) The above intermediate 1-(2-amino-4-bromo-5-fluorophenyl)-3-(4-methoxyphenyl)prop-2-yn-1-one (I e -04) was dissolved in 15 ml of diethylamine and stirred at room temperature overnight. After the reaction was completed, the reactant was washed with water and extracted with EA three times. The intermediate 7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I f -04). White solid, 2.4 g, yield 53.3%. 1 H NMR(600MHz,DMSO-d6)δ11.86(s,1H),8.60(s,1H),7.93(dd,J=10.8,8.8Hz,1 H),7.81(dd,J=18.8,6.7Hz,2H),7.17–7.12(m,2H),6.35(s,1H),3.85(s,3H).

[0166] (4) In a dry two-necked round-bottom flask, the above intermediate 7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I f-04) (100 mg), tris(dibenzylideneacetone)dipalladium(0) (0.05 equiv), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.05 equiv), and sodium tert-butoxide (2 equiv) were added and the reaction system was sealed. Then, 3 mL of toluene was added as a solvent. The air in the reaction system was displaced with nitrogen, and the N-ethylpiperazine raw material (1.2 equiv) was added dropwise to the reaction system with a syringe and sealed. The mixture was stirred at 120 °C for 4 - 5 h. After the reaction was completed, it was cooled to room temperature, and the catalyst was removed by suction filtration through diatomaceous earth. The filtrate was evaporated under vacuum to remove the solvent. Finally, the quinolone compound 7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-04) was purified by column chromatography (SiO2, DCM:MeOH = 5:1). White solid, 63 mg, yield 57.5%. 1 H NMR (400 MHz, CDCl3) δ 11.60 (s, 1H), 7.69 (d, J = 13.3 Hz, 1H), 7.54 (d, J = 8.3 Hz, 2H), 7.24 (d, J = 7.3 Hz, 1H), 6.81 (d, J = 8.3 Hz, 2H), 6.15 (s, 1H), 3.67 (s, 3H), 3.13 (t, J = 4.8 Hz, 4H), 2.69–2.46 (m, 4H), 2.38 (q, J = 7.2 Hz, 2H), 1.02 (t, J = 7.1 Hz, 3H); HRMS: calcd for C 22 H 24 FN3O2 [M + H] + , 382.1925; found 382.1921.

[0167] Example 5

[0168] Preparation of 1-benzyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-05):

[0169]

[0170] (1) While stirring, iodine (1.2 equiv, 60 mmol) was slowly added to a solution of 3-bromo-4-fluoroaniline raw material (1 equiv, 9.5 g) in 50 mL of deionized water. The reaction was continuously stirred at room temperature for 2 h. After completion, the reaction was stopped, and it was extracted with aqueous sodium thiosulfate solution and EA. Subsequently, the organic phase was dried over anhydrous sodium sulfate. The intermediate 5-bromo-4-fluoro-2-iodoaniline (I d -04) was purified by column chromatography (SiO2, PE:EA = 20:1). White solid, 8.8 g, yield 54%. 1H NMR (400MHz, CDCl3) δ7.42 (d, J = 7.6 Hz, 1H), 6.93 (d, J = 6.0 Hz, 1H), 4.01 (s, 2H).

[0171] (2) In a dry two-necked round-bottom flask, the intermediate (I d -04) (8.8 g), molybdenum hexacarbonyl (1.5 eq.), palladium acetate (0.02 eq.) and tri-tert-butylphosphine tetrafluoroborate (0.06 eq.) were dissolved in 15 ml of ultra-dry acetonitrile. The flask was sealed, the air was replaced with nitrogen, and stirred at room temperature. Triethylamine (2 eq.) and 4-ethynylanisole (1.2 eq.) were slowly added dropwise with a syringe. After stirring at room temperature for 16 hours, the mixture was extracted three times with saturated aqueous ammonium chloride solution and EA, dried over anhydrous sodium sulfate, and purified by column chromatography (SiO2, PE:EA=8:1) to obtain the pure intermediate 1-(2-amino-4-bromo-5-fluorophenyl)-3-(4-methoxyphenyl)prop-2-yn-1-one (I e -04). Yellow solid, 4.5 g, yield 46.3%. 1 H NMR (400MHz, CDCl3) δ7.91 (d, J = 9.2Hz, 1H), 7.68–7.62 (m, 2H), 6.99–6.90 (m, 3H), 6.26 (s, 2H), 3.89 (s, 3H).

[0172] (3) The above intermediate 1-(2-amino-4-bromo-5-fluorophenyl)-3-(4-methoxyphenyl)prop-2-yn-1-one (I e -04) was dissolved in 15 ml of diethylamine and stirred at room temperature overnight. After the reaction was completed, the reactant was washed with water and extracted with EA three times. The intermediate 7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I f -04). White solid, 2.4 g, yield 53.3%. 1 H NMR(600MHz,DMSO-d6)δ11.86(s,1H),8.60(s,1H),7.93(dd,J=10.8,8.8Hz,1 H),7.81(dd,J=18.8,6.7Hz,2H),7.17–7.12(m,2H),6.35(s,1H),3.85(s,3H).

[0173] (4) In a dry round-bottom flask, the above intermediate 7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I f(200 mg) of -04 was dissolved in 3 mL of ultra-dry N,N-dimethylformamide and stirred in an ice bath. After adding sodium hydride (1.2 equiv) to the mixture and stirring for 30 minutes, benzyl bromide (1.2 equiv) was added dropwise thereto. After the addition was complete, the reaction flask was removed from the ice bath. The reaction was carried out at 60 °C for 3 hours. After the reaction was completed, it was extracted 3 - 5 times with EA and brine to remove the solvent. Then the organic layer was concentrated in vacuo and purified by column chromatography (SiO2, DCM:MeOH = 40:1) to obtain the pure intermediate 1-benzyl-7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I g -05). White solid, 195.6 mg, yield 77.7%.

[0174] (5) In a dry two-necked round-bottom flask, the above intermediate 1-benzyl-7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I g -05) (195.6 mg), tris(dibenzylideneacetone)dipalladium(0) (0.05 equiv), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (0.05 equiv) and sodium tert-butoxide (2 equiv) were added and the reaction system was sealed. Then 3 mL of toluene was added as a solvent. The air in the reaction system was displaced with nitrogen, and the N-ethylpiperazine raw material (1.2 equiv) was added dropwise to the reaction system with a syringe and sealed. The reaction was stirred at 120 °C for 4 - 5 hours. After the reaction was completed, it was cooled to room temperature, and the catalyst was removed by suction filtration through diatomaceous earth. The filtrate was evaporated in vacuo to remove the solvent. Finally, it was purified by column chromatography (SiO2, DCM:MeOH = 10:1) to obtain the quinolone compound 1-benzyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-05). White solid, 148.78 mg, yield 70.7%. 1 H NMR (400 MHz, DMSO-d6) δ 8.22 (d, J = 8.6 Hz, 2H), 7.68 (d, J = 13.5 Hz, 1H), 7.60 (d, J = 7.5 Hz, 2H), 7.53 (s, 1H), 7.46 (t, J = 7.5 Hz, 2H), 7.39 (dd, J = 7.7, 2.5 Hz, 2H), 7.08 (d, J = 8.6 Hz, 2H), 5.51 (s, 2H), 3.85 (s, 3H), 3.18 (t, J = 4.8 Hz, 4H), 2.57 (t, J = 4.7 Hz, 4H), 2.41 (d, J = 7.2 Hz, 2H), 1.05 (t, J = 7.1 Hz, 3H); HRMS: calcd for C 29 H 30 FN3O2 [M + H] +,472.2395; found 472.2383.

[0175] Example 6

[0176] Preparation of 7-(4-ethylpiperazin-1-yl)-6-fluoro-1-(4-methoxybenzyl)-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-06):

[0177]

[0178] Except that the reaction raw material benzyl bromide in (4) of Example 5 was replaced with 4-methoxybenzyl bromide, the other required raw materials, reagents and preparation methods were the same as those in Example 5, and the total reaction yield was 73%. This compound is a yellow solid with a yield of 75%. 1 H NMR(400MHz,DMSO-d6)δ8.29–8.19(m,2H),7.63(d,J = 13.5Hz,1H),7.54(d,J = 8.2Hz,3H),7.40(d,J = 8.4Hz,1H),7.14–7.05(m,2H),7.06–6.98(m,2H),5.43(s,2H),3.86(s,3H),3.79(s,3H),3.18(s,4H),2.58(s,4H),2.42(d,J = 7.7Hz,2H),1.06(t,J = 7.1Hz,3H); HRMS:calcd forC 30 H 32 FN3O3[M+H] + ,502.2500; found 502.2493.

[0179] Example 7

[0180] Preparation of 7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)-1-(4-(trifluoromethyl)benzyl)quinolin-4(1H)-one (I-07):

[0181]

[0182] Except that the reaction raw material benzyl bromide in (4) of Example 5 was replaced with 4-(trifluoromethyl)benzyl bromide, the other required raw materials, reagents and preparation methods were the same as those in Example 5, and the total reaction yield was 73%. This compound is a white solid with a yield of 70.4%. 11H NMR (400 MHz, DMSO-d6) δ 8.25–8.19 (m, 2H), 7.82 (d, J = 1.7 Hz, 4H), 7.74 (d, J = 14.6 Hz, 1H), 7.53 (s, 1H), 7.40 (d, J = 8.5 Hz, 1H), 7.12–7.06 (m, 2H), 5.63 (s, 2H), 3.85 (s, 3H), 3.18 (s, 4H), 2.57 (s, 4H), 2.41 (q, J = 7.2 Hz, 2H), 1.10–1.01 (m, 3H); HRMS: calcd for C 30 H 29 F4N3O2 [M+H] + , 540.2269; found 540.2258。

[0183] Example 8

[0184] Preparation of 1-benzyl-7-(4-ethylpiperazin-1-yl)-2-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-6-carbonitrile (I-08):

[0185]

[0186] Except that the reaction raw material 3-bromo-4-fluoroaniline in (1) of Example 5 was replaced with 4-amino-2-bromobenzonitrile, the other required raw materials, reagents and preparation methods were the same as those in Example 5. This compound was a brown solid with a yield of 70.4%. 1 1H NMR (600 MHz, DMSO) δ 8.45 (s, 1H), 8.32–8.25 (m, 2H), 7.62 (d, J = 7.2 Hz, 3H), 7.49–7.44 (m, 3H), 7.42–7.36 (m, 1H), 7.14–7.08 (m, 2H), 5.56 (s, 2H), 3.86 (s, 3H), 3.25 (s, 4H), 2.61 (d, J = 8.3 Hz, 4H), 2.43 (d, J = 7.4 Hz, 2H), 1.06 (t, J = 7.2 Hz, 3H); HRMS: calcd for C 30 H 30 N4O2 [M+H] + , 479.2442; found 479.2430。

[0187] Example 9

[0188] Preparation of 1-benzyl-7-((3-chloropropyl)amino)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-09):

[0189]

[0190] Except that the reaction raw material N-ethylpiperazine in (5) of Example 5 was replaced with 3-chloropropylamine, the rest of the required raw materials, reagents and preparation methods were the same as those in Example 5. This compound was a white solid with a yield of 51.3%. 1 H NMR(400MHz,DMSO)δ8.22–8.13(m,2H),7.64–7.55(m,3H),7.45(t,J=7.5Hz,2H),7.38(d,J=6.5Hz,2H),7.09–7.04(m,2H),7.01(d,J=8.4Hz,1H),6.29(q,J=5.7Hz,1H),5.47(s,2H),3.88–3.75(m,5H),3.37(q,J=6.5Hz,2H),2.11(p,J=6.6Hz,2H);HRMS:calcd for C 26 H 24 ClFN2O2[M+H] + ,451.1583;found 451.1581.

[0191] Example 10

[0192] Preparation of 1-benzyl-6-fluoro-2-(4-methoxyphenyl)-7-(propylamino)quinolin-4(1H)-one (I-10):

[0193]

[0194] Except that the reaction raw material N-ethylpiperazine in (5) of Example 5 was replaced with propylamine, the rest of the required raw materials, reagents and preparation methods were the same as those in Example 5. This compound was a white solid with a yield of 63%. 1 H NMR(400MHz,DMSO)δ8.28–8.07(m,2H),7.58(dd,J=9.8,2.7Hz,3H),7.45(dd,J=8.3,6.7Hz,2H),7.41–7.33(m,2H),7.10–7.02(m,2H),6.96(d,J=8.4Hz,1H),6.17(dt,J=7.6,3.7Hz,1H),5.47(s,2H),3.84(s,3H),3.23–3.13(m,2H),1.67(h,J=7.4Hz,2H),0.98(t,J=7.4Hz,3H);HRMS:calcd forC 26 H 25 FN2O2[M+H] + ,417.1973;found 417.1961.

[0195] Embodiment 11

[0196] Preparation of 1-benzyl-7-(cyclopropylmethoxy)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-11):

[0197]

[0198] (1) While stirring, iodine (1.2 equivalents, 60 mmol) was slowly added to a solution of 3-bromo-4-fluoroaniline raw material (1 equivalent, 9.5 g) in 50 ml of deionized water. The reaction was stirred continuously at room temperature for 2 hours. After completion, the reaction was stopped and extracted with sodium thiosulfate aqueous solution and EA. Subsequently, the organic phase was dried over anhydrous sodium sulfate. The intermediate 5-bromo-4-fluoro-2-iodoaniline (I) was purified by column chromatography (SiO2, PE:EA=20:1). d -04). White solid, 8.8 g, yield 54%. 1 H NMR (400MHz, CDCl3) δ7.42 (d, J = 7.6 Hz, 1H), 6.93 (d, J = 6.0 Hz, 1H), 4.01 (s, 2H).

[0199] (2) In a dry two-necked round-bottom flask, the intermediate (I d -04) (8.8 g), molybdenum hexacarbonyl (1.5 eq.), palladium acetate (0.02 eq.) and tri-tert-butylphosphine tetrafluoroborate (0.06 eq.) were dissolved in 15 ml of ultra-dry acetonitrile. The flask was sealed, the air was replaced with nitrogen, and stirred at room temperature. Triethylamine (2 eq.) and 4-ethynylanisole (1.2 eq.) were slowly added dropwise with a syringe. After stirring at room temperature for 16 hours, the mixture was extracted three times with saturated aqueous ammonium chloride solution and EA, dried over anhydrous sodium sulfate, and purified by column chromatography (SiO2, PE:EA=8:1) to obtain the pure intermediate 1-(2-amino-4-bromo-5-fluorophenyl)-3-(4-methoxyphenyl)prop-2-yn-1-one (I e -04). Yellow solid, 4.5 g, yield 46.3%. 1 H NMR (400MHz, CDCl3) δ7.91 (d, J = 9.2Hz, 1H), 7.68–7.62 (m, 2H), 6.99–6.90 (m, 3H), 6.26 (s, 2H), 3.89 (s, 3H).

[0200] (3) The above intermediate 1-(2-amino-4-bromo-5-fluorophenyl)-3-(4-methoxyphenyl)prop-2-yn-1-one (I e-04) was dissolved in 15 mL of diethylamine and stirred overnight at room temperature. After the reaction was completed, the reactants were washed with water and extracted three times with EA. Purification by column chromatography (SiO2, DCM:MeOH = 5:1) gave the intermediate 7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I f -04). White solid, 2.4 g, yield 53.3%. 1 H NMR (600 MHz, DMSO-d6) δ 11.86 (s, 1H), 8.60 (s, 1H), 7.93 (dd, J = 10.8, 8.8 Hz, 1H), 7.81 (dd, J = 18.8, 6.7 Hz, 2H), 7.17–7.12 (m, 2H), 6.35 (s, 1H), 3.85 (s, 3H).

[0201] (4) In a dry round-bottom flask, the above intermediate 7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I f -04) (200 mg) was dissolved in 3 mL of ultra-dry N,N-dimethylformamide and stirred in an ice bath. Sodium hydride (1.2 equiv) was slowly added to the mixture and stirred for 30 minutes, then benzyl bromide (1.2 equiv) was added dropwise thereto. After addition, the reaction flask was removed from the ice bath. The reaction was carried out at 60 °C for 3 h. After the reaction was completed, the mixture was extracted 3–5 times with EA and brine to remove the solvent. Then the organic layer was concentrated in vacuo and purified by column chromatography (SiO2, DCM:MeOH = 40:1) to give the pure intermediate 1-benzyl-7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I g -05). White solid, 208.3 mg, yield 83%.

[0202] (5) In a dry two-necked round-bottom flask, the above intermediate 1-benzyl-7-bromo-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I g-05) (208.3 mg), 2-(di-tert-butylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl palladium(II) (2-amino-1,1'-biphenyl-2-yl) methanesulfonate (0.1 eq), and cesium carbonate (2 eq) were added and the reaction system was sealed. Then, 3 mL of toluene was added as a solvent. The air in the reaction system was displaced with nitrogen, and 1.2 eq of trifluoroethanol was added dropwise to the reaction system with a syringe and sealed. Subsequently, the mixture was stirred at 90 °C for 4 h. After the reaction was completed, it was cooled to room temperature, and the catalyst was removed by suction filtration through diatomaceous earth. The filtrate was evaporated under vacuum to remove the solvent. Finally, it was purified by column chromatography (SiO2, PE:EA = 10:1) to obtain the intermediate 6-fluoro-7-hydroxy-2-(4-methoxyphenyl)quinolin-4(1H)-one (I h -11). Yellow solid, 104 mg, yield 61%.

[0203] (6) In a dry round-bottom flask, the intermediate 6-fluoro-7-hydroxy-2-(4-methoxyphenyl)quinolin-4(1H)-one (I h -11) (104 mg) was dissolved in 3 mL of anhydrous N,N-dimethylformamide. Subsequently, the starting material (iodomethyl)cyclopropane (1.2 eq) and 2 eq of cesium carbonate were added, and the mixture was stirred at room temperature. After the reaction was completed, it was extracted 3-5 times with EA and brine to remove the solvent. Then, the organic layer was concentrated under vacuum and purified by column chromatography (SiO2, DCM:MeOH = 40:1) to obtain the quinolone compound 1-benzyl-7-(cyclopropylmethoxy)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-11). 1 H NMR (400 MHz, DMSO) δ 8.27–8.17 (m, 2H), 7.76 (d, J = 11.8 Hz, 1H), 7.64–7.54 (m, 3H), 7.53–7.44 (m, 3H), 7.42–7.35 (m, 1H), 7.12–7.04 (m, 2H), 5.52 (s, 2H), 4.08 (d, J = 7.1 Hz, 2H), 3.85 (s, 3H), 1.33 (t, J = 7.6 Hz, 1H), 0.67–0.60 (m, 2H), 0.45–0.38 (m, 3H); HRMS: calcd for C 27 H 24 FNO3 [M + H] + , 430.1813; found 430.1806.

[0204] Example 12

[0205] Preparation of 1-benzyl-6-fluoro-7-methoxy-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-12):

[0206]

[0207] Except that the reaction raw material (iodomethyl) cyclopropane in (6) of Example 11 was replaced with iodomethane, the other required raw materials, reagents and preparation methods were the same as those in Example 12. This compound was a white solid with a yield of 64%. 1 H NMR (400 MHz, DMSO) δ 8.28–8.19 (m, 2H), 7.76 (d, J = 11.9 Hz, 1H), 7.63–7.53 (m, 4H), 7.49–7.43 (m, 2H), 7.42–7.35 (m, 1H), 7.13–7.04 (m, 2H), 5.53 (s, 2H), 4.02 (s, 3H), 3.85 (s, 3H); HRMS: calcd for C 24 H 20 FNO3[M+H] + , 390.1500; found 390.1490.

[0208] Example 13

[0209] Preparation of 1-benzyl-6-fluoro-7-isopropoxy-2-(4-methoxyphenyl)quinolin-4(1H)-one (I-13):

[0210]

[0211] Except that the reaction raw material (iodomethyl) cyclopropane in (6) of Example 11 was replaced with 2-iodopropane, the other required raw materials, reagents and preparation methods were the same as those in Example 12. This compound was a white solid with a yield of 65%. 1 H NMR (400 MHz, DMSO) δ 8.29–8.17 (m, 2H), 7.75 (d, J = 11.9 Hz, 1H), 7.64–7.57 (m, 2H), 7.54 (d, J = 7.4 Hz, 2H), 7.50–7.42 (m, 2H), 7.41–7.33 (m, 1H), 7.12–7.05 (m, 2H), 5.52 (s, 2H), 4.93 (p, J = 6.0 Hz, 1H), 3.85 (s, 3H), 1.39 (d, J = 6.0 Hz, 6H); HRMS: calcd for C 26 H 24 FNO3[M+H] + , 418.1813; found 418.1800.

[0212] Biological experiment

[0213] In vitro antibacterial activity experiment of compounds I-01 to I-13 in Example 14

[0214] This example is an experiment for determining the minimum inhibitory concentration (MIC) of the compounds described in the present invention in vitro, and the antibacterial activity of the compounds was evaluated using the MIC value.

[0215] Experimental method: The in vitro antibacterial activity against Escherichia coli OP50-1 derivatives was evaluated by the microbial dilution method. Stock solutions of all derivatives were uniformly prepared with DMSO at a concentration of 51.2 mg / mL. A single colony of Escherichia coli OP50-1 was inoculated into LB liquid medium (Luria-Bertani Broth), and then shaken at 250 rpm at 37 °C for 12-18 hours on a shaker. After the bacterial solution became turbid, the bacterial solution was diluted with LB to OD 600 = 0.001. An equal volume of the OP50-1 bacterial solution was added to the wells to be tested, and then the stock solution of the compound was added to the wells to be tested and serially diluted by half in gradient. Then, they were cultured together in an incubator at 37 °C for 18 hours. The MIC value was defined as the minimum drug concentration at which no bacterial growth occurred. The OP50-1 solution without drug administration and pure LB were used as positive or negative controls, respectively.

[0216] Experimental results: The antibacterial activity data are shown in Table 1. The MIC value of the lead compound enrofloxacin was 0.0078 mg / L, thus confirming its extremely strong antibacterial activity; while for the modified compounds I-01 to I-03, I-09 to I-13, the MIC values against OP50-1 were all greater than 1024 mg / L, so it could be shown that the said compounds completely lost their inhibitory effect on it; the MIC values of I-04 to I-08 against OP50-1 were 512 mg / L, indicating that compounds I-04 to I-08 had a slight inhibitory effect on it.

[0217] Table 1

[0218]

[0219] Caenorhabditis elegans lifespan experiment of non-antibacterial compounds I-01 to I-03, I-09 to I-13 in Example 15

[0220] In this example, through the nematode lifespan experiment, the anti-aging activity of compounds was evaluated by the lifespan curves drawn after administering non-antibacterial compounds to nematodes. Since the lead compound enrofloxacin and compounds I-04 to I-08 all have a certain degree of antibacterial activity, firstly, it does not meet the purpose of structural modification to remove antibacterial activity; secondly, the antibacterial activity of the drug will cause a reduction in the food bacterial solution of nematodes, thereby triggering the dietary restriction effect to promote anti-aging effects. Therefore, using antibacterial compounds to conduct nematode lifespan experiments will cause great deviations in the experimental results and cannot reflect the true anti-aging effects of the compounds. Therefore, in this example, nematode lifespan experiments were carried out on non-antibacterial compounds I-01 to I-03, I-09 to I-13 to compare their anti-aging activities.

[0221] Experimental method: At 20 °C, synchronized L1 nematodes were cultured on 60-mm NGM plates for 36 - 48 hours until the L4 stage, during which sufficient Escherichia coli OP50-1 was seeded on the medium as food. Subsequently, L4 adult nematodes were transferred to blank or 35-mm NGM plates containing the compound (the initial screening drug concentration was uniformly 64 mg / L; the rescreening drug concentration was 128 mg / L). In addition, each plate also contained 50 μg / mL of FUdR to inhibit their reproduction (about 15 - 20 nematodes per plate); the drug administration period lasted from day 0 to day 9. The surviving nematodes were counted under a microscope every 2 days and transferred to fresh NGM plates. The death of nematodes was evaluated as no response to any repeated mechanical stimulation. From day 10 until all samples died, the NGM of all groups was replaced with NGM plates without any drug or FUdR. The survival curves of nematodes were obtained by plotting with GraphPad Prism 8.3.0 software, and their significance was evaluated by Log-rank.

[0222] Experimental results: The initial screening of lifespan extension activity of nematodes is shown in Table 2. The low-concentration initial screening data of the compounds showed that compounds I-01 to I-03, I-09 to I-13 all have a certain degree of lifespan extension activity for nematodes; for the compounds I-01, I-02, and I-03 with the best activity, high-concentration rescreening of the same batch was continued. The results are shown in Table 3. It was found that the activity of I-02 was the best. Therefore, the pharmacodynamic determination of compound I-02 at the subsequent nematode health lifespan, cell level, and mouse level was carried out.

[0223] Table 2 Initial screening data of nematode lifespan for compounds I-01 to I-03, I-09 to I-13

[0224]

[0225] Table 3 Rescreening data of nematode lifespan for compounds I-01 to I-03

[0226]

[0227] Evaluation of the healthy lifespan of nematodes with the preferred compound I-02 in Example 16

[0228] The healthy lifespan of nematodes in this example was evaluated using pharyngeal pumping rate, swimming rate, and osmotic stress resistance ability as evaluation indicators.

[0229] Experimental method:

[0230] (1) Pharyngeal pumping experiment: Under the same conditions as in the lifespan determination, the nematodes were cultured to adulthood. At the L4 stage, the nematodes were transferred to NGM plates with or without I-02 (at concentrations of 64 mg / L and 128 mg / L). On the 4th, 8th, and 12th days, their pharyngeal pumping rates were counted within 30 seconds using a stereomicroscope. Approximately 15 to 20 samples were counted in each group.

[0231] (2) Swimming experiment: The nematodes at the L4 stage were transferred to NGM plates with or without I-02 (at concentrations of 64 mg / L and 128 mg / L). The body bending speed of the samples was counted on the 3rd, 8th, and 12th days. Each sample was transferred to M9 buffer for 30 seconds to adapt. Then, the body swings were counted within 30 seconds. Approximately 15 to 20 samples were counted in each group.

[0232] (3) Osmotic stress experiment: The nematodes at the L4 stage were transferred to NGM plates and cultured for 4 days with or without I-02 (64 mg / L, 128 mg / L). On the 5th day, all plates were replaced with 500 mM NaCl. The duration of paralysis of all samples was recorded, and the survival curve under high osmotic pressure conditions was plotted.

[0233] Experimental results: The results of the I-02 nematode lifespan experiment are as Figure 1 shown. Compared with the blank group, I-02 administration could increase the pharyngeal pumping rate and swimming rate of nematodes. In the osmotic stress experiment, the survival rate of the I-02 administration group was significantly higher than that of the blank control, indicating that the compound of the present invention can improve the ability to resist high osmotic pressure environments.

[0234] Evaluation of the efficacy of the preferred compound I-02 on senescent cells in Example 17

[0235] The efficacy evaluation in this example includes two senescent cell line models: the replicative human embryonic lung fibroblast MRC-5 senescent cell model and the Mito-C-induced rat renal tubular epithelial cell NRK-52E senescent cell model; among which the detection of senescence indicators includes the detection of cell cycle arrest-related proteins (p16, p21, p53, etc.), the DNA damage marker γ-H2AX, and senescence-associated secretory phenotype factors (SASP factors, such as MMP-3, ICAM, CXCL-1, CXCL-10, CCL-2, IL-6, and IL-8), etc.

[0236] Experimental method:

[0237] (1) Replicative MRC-5 senescent cells: The cells were normally passaged to the senescent generations P28 - P30.

[0238] (2) Mito-C-induced NRK-52E senescent cells: Normal NRK-52E cells were modeled with 0.5 μM Mito-C for 48 hours, and then changed to normal medium and left standing for 48 hours.

[0239] (3) Administer I-02 (20 μM) to the above two types of cells for 48 hours.

[0240] (4) Detection of cell cycle arrest-related proteins: Western blot was used to detect the changes in the corresponding protein levels before and after drug administration.

[0241] (5) Detection of SASP factors: q-PCR method was used to detect the changes in the corresponding gene levels before and after drug administration.

[0242] Experimental results: The efficacy results of I-02 on senescent cells are as Figure 2 shown. I-02 can significantly reduce the expression of senescence-related cell cycle arrest proteins p21 and p53 in the two types of senescent cells; moreover, I-02 can significantly reduce the expression of SASP factors in replicative senescent MRC-5 cells, acting as a Senomorphic drug to intervene in the cell senescence process.

[0243] Efficacy evaluation of the preferred compound I-02 in doxorubicin-induced senescent mice in Example 18

[0244] The efficacy index evaluation in this example includes the detection of the senescence marker β-galactosidase SA-β-gal in kidney sections and the detection of kidney senescence-associated secretory phenotype factors (SASP factors, such as CXCL-1, CXCL-3, IL-1β, IL-6, MMP-1, MMP-7, and TNF-α).

[0245] Experimental method: The lead compounds enrofloxacin and I-02 (5 mg / kg and 20 mg / kg) were completely dissolved in a mixture of 10% DMSO, 40% PEG400 and 50% water, metformin (20 mg / kg) was completely dissolved in water, and a 5 mg / kg doxorubicin solution was prepared with water. On day 0 and day 10, male C57BL / 6j mice were induced to premature senility by intraperitoneal injection of 5 mg / kg doxorubicin. After normal feeding for 5 days, the mice were treated by oral gavage every day. The mice were sacrificed on day 38, and samples were taken to detect physiological and biochemical indexes. The detection method of SA-β-gal in kidney sections was immunohistochemistry; the detection method of SASP factors in the kidney was ELISA.

[0246] Experimental results: The in vivo pharmacodynamic results of I-02 in doxorubicin-induced premature senility mice are as Figure 3 shown. Compared with the model group, 20 mg / kg I-02 could significantly reduce the expression of SA-β-gal, and was superior to the positive drug metformin at the same dose; the expression results of SASP factors in the kidney showed that compared with the model group, I-02 could significantly reduce the expression of SASP factors in the kidneys of mice at both 5 mg / kg and 20 mg / kg doses, and the treatment effects were better than those of the lead compound enrofloxacin at the same dose.

[0247] In summary, in order to avoid the side effects such as drug resistance and intestinal flora imbalance caused by long-term use of antibacterial drugs, the compound of the present invention was prepared. The compound showed excellent anti-aging activity in vitro and in vivo, and there was no report on the treatment of similar compounds at present, providing a new drug use mode for the development of anti-aging drugs and drugs for the treatment of aging-related diseases.

[0248] All documents mentioned in the present invention are incorporated herein by reference as if each document was individually incorporated by reference. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the appended claims of the present application.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt thereof, wherein, R 1 selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C2-C6 alkenyl, substituted or unsubstituted C2-C6 alkynyl, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, substituted or unsubstituted benzyl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S, substituted or unsubstituted C1-C6 alkyl-5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; R 2 selected from the group consisting of: substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C6-C10 aryl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S; R 6 selected from the group consisting of: H, halogen, -OH, -NH2, -NO2, -CN, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl; R 7 selected from the group consisting of: -NR 10 and -OR 10 ; Each R 10 is independently selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkyl, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl; or two Rs 10 are linked to form a substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-2 heteroatoms selected from N, O or S; each substitution independently refers to being substituted by 1 - 3 substituents selected from the group consisting of: -OH, -NH2, -NO2, -CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, C6-C10 aryl, 5-7 membered heteroaryl containing 1 - 3 heteroatoms selected from N, O or S.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, R 1 selected from the group consisting of: H, substituted or unsubstituted C1-C6 alkoxy, substituted or unsubstituted C3-C8 cycloalkyl, substituted or unsubstituted C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, substituted or unsubstituted benzyl, substituted or unsubstituted 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S, substituted or unsubstituted C1-C6 alkyl-5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S.

3. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compound of formula I is the compound of formula II: wherein, R 2 , R 6 , R 7 and R 10 are as defined in claim 1; Each R 11 is independently selected from the group consisting of: H, -OH, -NH2, -NO2, -CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, C3-C8 heterocycloalkyl containing 1-3 heteroatoms selected from N, O or S, C6-C10 aryl, 5-7 membered heteroaryl containing 1-3 heteroatoms selected from N, O or S.

4. The compound according to claim 3, or a pharmaceutically acceptable salt thereof, wherein Each R 11 is independently selected from the group consisting of: H, -OH, -NH2, -NO2, -CN, halogen, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 alkoxy, C1-C6 haloalkoxy, C3-C8 cycloalkyl, C6-C10 aryl.

5. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, The compounds are selected from the group consisting of: 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-phenylquinolin-4(1H)-one; 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-cyclopropyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(thiophen-2-yl)quinolin-4(1H)-one; 7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 7-(4-ethylpiperazin-1-yl)-6-fluoro-1-(4-methoxybenzyl)-2-(4-methoxyphenyl)quinolin-4(1H)-one; 7-(4-ethylpiperazin-1-yl)-6-fluoro-2-(4-methoxyphenyl)-1-(4-(trifluoromethyl)benzyl)quinolin-4(1H)-one; 1-benzyl-7-(4-ethylpiperazin-1-yl)-2-(4-methoxyphenyl)-4-oxo-1,4-dihydroquinoline-6-carbonitrile; 1-benzyl-7-((3-chloropropyl)amino)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-6-fluoro-2-(4-methoxyphenyl)-7-(propylamino)quinolin-4(1H)-one; 1-benzyl-7-(cyclopropylmethoxy)-6-fluoro-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-6-fluoro-7-methoxy-2-(4-methoxyphenyl)quinolin-4(1H)-one; 1-benzyl-6-fluoro-7-isopropoxy-2-(4-methoxyphenyl)quinolin-4(1H)-one.

6. A method for preparing the compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, Comprising the following steps: In an inert solution, the compound of formula I c reacts with the H-R 7 compound by a nucleophilic substitution reaction to thereby prepare the compound of formula I; wherein each group is as defined in claim 1.

7. A pharmaceutical composition, characterized in that, Comprising: (i) The compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof; and (ii) A pharmaceutically acceptable carrier.

8. Use of the compound according to claim 1, or a pharmaceutically acceptable salt thereof, characterized in that, For the preparation of a medicament, said medicament (a) for extending lifespan; (b) for delaying aging; (c) for treating aging-related diseases.

9. The use according to claim 8, characterized in that, Said diseases are selected from the group consisting of: tumors, acute or chronic renal failure, liver aging, Alzheimer's disease, heart failure, neurodegenerative diseases, hypertension, diabetes.

10. A method for delaying and / or treating aging, characterized in that, Comprising the step of administering to a subject the compound as claimed in claim 1, or a pharmaceutically acceptable salt thereof or the composition as claimed in claim 7.

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