WO2018233483A1 - Preparation and use of phenylquinolinone and flavonoid derivatives - Google Patents

Abstract

Provided are phenylquinolinone and flavonoid derivatives and preparation methods therefor, wherein the derivatives are histamine H3 receptor antagonists, which can protect and restore the normal function of the central nervous system; and in particular provided is the use thereof in neurodegenerative disease and ischemic brain injury. The phenylquinolinone and flavonoid derivatives have great clinical application value.

Description

Preparation and application of phenylquinolinones and flavonoid derivatives Technical field

The invention belongs to the technical field of medicine, and particularly relates to the preparation of phenylquinolinones and flavonoid derivatives, wherein the active ingredient is a histamine H3 receptor antagonist, which can protect and restore the normal function of the central nervous system, and in the neurodegenerative Application in the treatment of sexual diseases, ischemic gas-deficient encephalopathy, Parkinson's syndrome, narcolepsy, epilepsy and gradual freezing diseases. The phenylquinolinones and flavonoid derivatives also include pharmaceutically acceptable salts, complexes, solvates thereof and the like.

Background technique

Histamine H ₃ receptor is present in the cerebral cortex, striatum, hippocampus, olfactory bulb, terminal nucleus, thalamus, low brainstem nucleus, cerebellum, etc. Histamine H ₃ receptor is the neuronal synapse itself. Exciting the receptor inhibits the release of histamine or other neurotransmitters, and antagonizing the receptor promotes neurotransmitter release.

Cerebral ischemia is mainly caused by thrombosis, embolism or other causes of cerebral infarction caused by insufficient blood supply to the brain. It is also one of the number one killers of human disability and death. Due to the complex mechanism of the pathological process, no effective drug has been applied to the clinic so far.

Studies have shown that histamine H ₃ receptor antagonists can be used to treat Alzheimer's disease (AD), Parkinson's disease (PD), and gradual freezing by promoting the synthesis and release of neurotransmitters such as histamine. ALS), narcolepsy and more.

Among them, Alzheimer's disease (AD) has been found for more than 100 years, and the number of cases has increased year by year. Studies have shown that β-amyloid (Aβ) aggregation, tau protein hyperphosphorylation, mitochondrial dysfunction, oxidation Many factors such as stress and loss of function of the neurotransmitter system may be related to its pathogenesis, and different molecular mechanisms are related to each other and affect each other. At present, the first-line therapeutic drugs for AD are mainly acetylcholinesterase (AChE) inhibitors donepezil, rivastigmine, galantamine and non-competitive NMDA receptor antagonist memantine, which all act on Cognitive-related neurotransmitter systems, while ameliorating cognitive impairment in early-stage patients, do not fundamentally improve disease status or halt disease progression.

The process of occurrence and development of AD disease involves a variety of factors, and the ability to simultaneously target multiple target-directed ligands (MTDLs) has unique advantages and has attracted widespread attention. Studies have shown that H ₃ receptors are important receptors for the regulation of neurotransmitters in the brain, and H ₃ receptor antagonists can effectively promote histamine, acetylcholine and other cognitive-related neurotransmitters by blocking H ₃ receptors. Release, currently, several histamine H ₃ receptor antagonists (ABT-288, AZD5213, CEP-26401, GSK239512 and MK0249) have entered clinical studies for Alzheimer's disease, Parkinson's disease, narcolepsy, etc. Treatment of neurodegenerative diseases

Figure 1 Enteres part of the H3 receptor antagonist in clinical studies.

Senile plaques (SP), which are deposited on the edge of the brain and in the cerebral cortex after aggregation of amyloid peptide Aβ, are one of the most typical pathological features of AD. Moreover, the oligomers, fibrils and fibers formed by Aβ during aggregation can be toxic to nerve cells and even cause neuronal apoptosis. Therefore, Αβ aggregation is generally considered to be the intrinsic cause of AD, and drugs that inhibit Aβ aggregation have become an important direction for the development of AD drugs.

The present invention systematically analyzes the biological characteristics of AD-related targets, organically combines a target for improving symptoms (H ₃ receptor) and a target for disease (Aβ aggregation), and uses a rational drug design method. Designing multi-target compounds that simultaneously antagonize H ₃ receptors and inhibit Aβ aggregation. These multi-target compounds not only promote the release of neurotransmitters, improve the symptoms of AD patients, delay the disease, but also inhibit Aβ. Self-aggregation can fundamentally prevent the progression of AD and achieve the effect of "multi-pronged approach, treating both the symptoms and the symptoms", and may have a unique effect on the treatment of neurodegenerative diseases such as AD.

The invention also discloses the protective effect of the compound of the invention on the neurological function of ischemic brain injury.

The molecular structure of histamine H ₃ receptor antagonists is mostly chain-like molecules, which are mainly composed of three parts: the western part is a nitrogen-containing basic region of a fatty tertiary amine and a part of a connecting chain, and the center is a fat-soluble aromatic ring, and the east is Fragments are fat-soluble aromatic rings or basic groups (Figure 2)

Figure 2 Target molecular design.

The present invention is based on the H ₃ receptor antagonist pharmacophore model, using benzoquinolinone (benzopyrone) as a fat-soluble aromatic ring (eastern fragment) and phenoxypropylamine as the other part (central ring) + western segment), designed and synthesized new phenylquinolinones and flavonoid derivatives; on the other hand, previous studies have shown that N-aminopropoxyphenyl-pyridin-4-one derivatives with similar structure It has good inhibition of Aβ aggregation activity (N-substituted phenylpyridin-4-one derivative and its preparation and application, ZL201310511154.2). Further tests showed that the activity of the designed phenyl quinolone derivatives and flavonoids H ₃ receptor antagonist having two targets and inhibitory compound Aβ aggregation.

Summary of the invention

Description of terms: As used herein, the term "alkyl", unless specified to indicate a different number of atoms, refers to a straight or branched hydrocarbon chain containing from 1 to 6 carbon atoms. Examples of "alkyl" as used in the present invention include, but are not limited to, methyl, ethyl, n-propyl, isobutyl, isopropyl. "Alkyl" also includes substituted alkyl groups. The alkyl group may be optionally substituted one or more times with a halogen or a hydroxyl group.

The term "alkoxy" as used herein, refers to an -O-alkyl group, wherein alkyl is as defined above. Examples of "alkoxy" as used herein include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy and tert-butoxy. "Alkoxy" also includes substituted alkoxy groups. The alkoxy group can be optionally substituted one or more times with a halogen.

The "pharmaceutically acceptable salts" in the present invention include hydrochlorides, sulfates, hydrobromides, (+) tartrates, (-) tartrates, fumarates, succinates, and the like which are commonly used in pharmacy. Inorganic acid salts and organic acid salts, amino acid salts.

The benzoquinolinones, flavonoids and analogs thereof, or pharmaceutically acceptable salts and solvates thereof, provided by the present invention have the following general structure (A):

General structure (A)

In the formula:

X is selected from N or O;

R _{1 is} selected from H, C _1-3 alkyl or C _1-3 alkoxy;

R _{2 is} selected from H, hydroxy or C _1-3 alkoxy;

R _{3 is} selected from H, hydroxy, C _1-3 alkyl or C _1-3 alkoxy.

NR'R" is selected from cyclic amines and open-chain alkylamines having a total of 3 to 6 carbon atoms, including but not limited to the following fragments:

Further, preferred quinolinones and flavonoid derivatives of the invention, said

Selected from the following two heterocycles:

Further, preferred quinolinones and flavonoid derivatives and analogs thereof of the invention, R ₁ is preferably hydrogen, methyl and ethyl; R ₂ is preferably hydrogen, hydroxy and methoxy; R ₃ is preferably hydrogen , hydroxyl and methoxy.

It will be understood that the present invention includes all combinations and subgroups of the specific groups defined by the present invention, including those as defined in the above summary, as illustrated in the various examples throughout the specification, and in the appended claims base.

More specifically, preferred compounds of the formula quinolinones and flavonoid derivatives of the present invention (or pharmaceutically acceptable salts, complexes, solvates of the compounds) are selected from the compounds shown in Table 1.

Table 1 Preferred compounds of quinolinone derivatives

The preparation method of the above compound provided by the present invention is prepared by the following procedure, but is not limited to the following method.

The quinolinone derivatives of the formula can be synthesized by the following steps.

Synthesis method of I-1 and I-2 in the compound I (quinolinone derivative) series (this method is suitable for X=N):

The specific reaction process is as follows: the raw material 1, 1,3-bromochloropropane and potassium carbonate are dissolved in acetonitrile and heated to reflux, and then reacted in a NaOH/CH ₃ OH system to obtain an intermediate 1 and then in a thionyl chloride solution. Intermediate 2 is refluxed, and then it is reacted with o-aminoacetophenone at room temperature to obtain Intermediate 3, which is reacted with potassium t-butoxide in a microwave to obtain Intermediate 4, and finally reacted with a secondary amine to obtain the target compound I- 1, I-2.

Synthesis method of I-3 to I-6 in the compound I (quinolinone derivative) series (this method is suitable for X=N):

The specific reaction process is as follows: the intermediate 4 in the first embodiment is dissolved in DMF, sodium hydride and a halogenated hydrocarbon are added to obtain the intermediate 5, and finally reacted with a secondary amine and triethylamine to obtain the target compound I-3. ~I-6.

Synthesis method of I-7 and I-8 in the compound I (quinolinone derivative) series (this method is suitable for X=N):

The specific reaction process is as follows: p-hydroxyacetophenone (raw material 4) is dissolved in acetonitrile, and 1,3-bromochloropropane and potassium carbonate are added to reflux to obtain intermediate 6, which is then reacted with bromine in an ice bath to obtain bromine. Ketone intermediate 7; the latter is reacted with anthranilic acid and potassium carbonate in DMF to obtain ester intermediate 8, which is refluxed with ammonium acetate/acetic acid to give intermediate 9 and finally refluxed with secondary amine and triethylamine. The target compound I-7, I-8 was obtained.

Synthesis method of I-9 and I-10 in the compound I (quinolinone derivative) series (this method is suitable for X=N):

The specific reaction process is as follows: the intermediate 9, the dimethyl sulfate and the potassium carbonate in the third embodiment are dissolved in acetone, and the intermediate is obtained by refluxing, and then reacted with the secondary amine NHR'R" and triethylamine. Target compound I-9, I-10.

Synthesis method of II-1, II-2 in compound II (flavonoid derivative) series (this method is suitable for X=O):

The specific reaction process is as follows: the raw material 6 is dissolved in acetonitrile, and after adding 1,3-bromochloropropane and potassium carbonate, the reaction is refluxed to obtain an intermediate 11 which is condensed with p-hydroxyacetophenone in potassium hydroxide. The intermediate 12 is then cyclized to give the intermediate 13 through a hydrogen peroxide / KOH system, and finally reacted with a second amine and triethylamine to give the object compound II-1, II-2.

Method for synthesizing II-3 and II-4 in the compound II (flavonoid derivative) series (this method is suitable for X=O)

The specific reaction process is as follows: the intermediate 13 in Example 5, methyl iodide and potassium carbonate are dissolved in acetone, and the intermediate 14 is refluxed, and then reacted with a secondary amine and triethylamine to obtain the target compound II-3. II-4.

Method for synthesizing II-5 and II-6 in compound II (flavonoid derivative) series (this method is suitable for X=O)

The specific reaction process is as follows: the intermediate 11 and 2,6-dihydroxyacetophenone are dissolved in ethanol, and after adding potassium hydroxide, the reaction is refluxed to obtain the intermediate 15, and then the cyclization reaction is carried out under the action of iodine and concentrated sulfuric acid. Intermediate 16 is finally reacted with a secondary amine or triethylamine to give the target compound II-5, II-6.

Method for synthesizing II-7 and II-8 in the compound II (flavonoid derivative) series (this method is suitable for X=O)

The specific reaction process is as follows: Example 7, intermediate 16, potassium iodide and potassium carbonate are dissolved in acetone, and refluxed to obtain intermediate 17, which is then refluxed with a secondary amine or triethylamine to obtain the target compound II-7, II-8. .

Another object of the present invention is to provide the use of the quinolinones, flavonoid derivatives and pharmaceutically acceptable salts and solvates thereof for the preparation of a medicament derived from the analogs and pharmaceutically Acceptable salts and solvates are prepared with pharmaceutically acceptable carriers or excipients.

The "pharmaceutically acceptable carrier" means a conventional pharmaceutical carrier in the pharmaceutical field, including conventional diluents in the pharmaceutical field, excipients such as water, etc., fillers such as starch, etc., binders such as cellulose derivatives, gelatin, etc. , wetting agents such as glycerin, disintegrating agents such as agar, calcium carbonate, etc., absorption enhancers such as quaternary ammonium compounds, surfactants such as cetyl alcohol, adsorption carriers such as kaolin and soap clay, lubricants such as talc, etc., if necessary Flavoring agents, sweeteners and the like can also be added.

The pharmaceutical preparations are suitable for administration by any appropriate route, such as oral (including buccal or sublingual administration), rectal administration, nasal administration, topical administration (including buccal, sublingual or transdermal administration). Or a parenteral administration (including subcutaneous, intramuscular, intravenous or intradermal) routes. These formulations can be prepared by any method known in the art of pharmacy. For example by a method of mixing the active ingredient with a carrier or excipient.

The present invention provides the compound and preferably a compound thereof, a pharmaceutically acceptable salt of the compound, a solvate of the compound, a prodrug (ester or phosphate), a stereoisomer, a deuterated substance, and the like The use of a combination of drugs in the treatment of neurodegenerative diseases. Wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, narcolepsy, epilepsy, gradual freezing, etc., in particular, the protection of the compound of the present invention and its preferred compounds against ischemic brain damage The effect is not only beneficial to the recovery of nerve function, but also effectively reduces the volume of cerebral infarction.

Detailed ways

The specific embodiments are intended to be illustrative, and are not to be construed as limiting. In addition, it should be understood that various changes and modifications may be made by those skilled in the art in the form of the present invention.

Example 1: Synthesis of I-1, I-2 in a class I compound (quinolinone).

Step 1: 4-(3-Chloropropoxy)benzoic acid (Intermediate 1).

Ethyl p-hydroxybenzoate II-1 (12 g, 72 mmol), 1,3-bromochloropropane (14.2 mL, 144 mmol) and K ₂ CO ₃ (20 g, 144 mmol) were dissolved in 100 mL of acetonitrile and heated to reflux for 12 h. Excess K ₂ CO _{3 was} removed by suction filtration, and the solvent was evaporated under reduced pressure to give a white oil. 15 mL of 6N NaOH solution and 30 mL of CH ₃ OH were added directly and refluxed for 1 h. The reaction mixture was clarified, cooled, acidified to pH = 2 with 2N hydrochloric acid, and a large white solid was precipitated, filtered, washed with water, and dried to give a white solid powder (14.5 g). Yield 94%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.08 (d, J = 8.5 Hz, 2H), 6.96 (d, J = 8.5 Hz, 2H), 4.21. (t, J = 6.0 Hz, 2H), 3.77 (t, J = 6.5 Hz, 2H), 2.30-2.25 (m, 2H); ESI-MS: m/z = 215 [M+H] ⁺ .

Step 2: N-(2-Acetylphenyl)-4-(3-chloropropoxy)benzamide (Intermediate 3).

Intermediate 1 (5.0g, 23mmol) in 10mL SOCl ₂ was refluxed for 1h, the reaction system was added to 2 1 drop of DMF. After completion of the reaction, excess SOCl ₂ was removed under reduced pressure to give a colorless liquid intermediate. The o-aminoacetophenone (2.83 g, 21 mmol) was dissolved in 15 mL of anhydrous CH ₂ Cl ₂ and 6.5 mL of anhydrous TEA, and the intermediate 2 was slowly added dropwise at 0 ° C, and the reaction was continued at room temperature for 2 h, suction filtration. The filtrate was dried with EtOAc (EtOAc:EtOAc) Yield 72%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 12.65 (s, 1H), 8.11 (d, J = 9.0 Hz, 1H), 8.05 (d, J = 9.0 Hz, 2H), 7.97 ( Dd, J = 8.0, 1.5 Hz, 1H), 7.64 (t, J = 8.0 Hz, 1H), 7.16 (t, J = 8.0 Hz, 1H), 7.02 (d, J = 9.0 Hz, 2H), 4.21 ( t, J = 6.0 Hz, 2H), 3.78 (t, J = 6.5 Hz, 2H), 2.73 (m, 3H), 2.31-2.26 (m, 2H); ESI-MS: m/z = 332 [M+ H] ⁺ .

Step three: 2-(4-(3-Chloropropoxy)phenyl)quinolin-4(1H)-one (Intermediate 4).

Intermediate 3 (995 mg, 3.0 mmol) and potassium tert-butoxide (1.68 g, 15 mmol) were dissolved in 15 mL THF and then reacted in a closed vessel at 110 ° C for 20 min. After the reaction was completed, it was cooled to room temperature, poured into 100 mL of ice water, adjusted to pH 5-6 with 2N HCl, and a large amount of yellow solid was precipitated, washed with water, and washed with a small amount of ice acetone and CH ₂ Cl ₂ (1:1) to obtain intermediate 4 , 750mg. Yield 80%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 11.63 (s, 1H), 8.10 (dd, J = 8.0, 1.0 Hz, 1H), 7.83 (d, J = 9.0 Hz, 2H), 7.78 (d, J = 8.0, 1H), 7.68 (t, J = 7.5 Hz, 1H), 7.35 (t, J = 7.5 Hz, 1H), 7.17 (d, J = 9.0 Hz, 2H), 6.33 (s) , 1H), 4.21 (t, J = 6.0 Hz, 2H), 3.84 (t, J = 6.5 Hz, 2H), 2.24 - 2.18 (m, 2H); ESI-MS: m/z = 314 [M+H ] ⁺ .

Step 4: 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)quinolin-4(1H)-one (I-1).

Intermediate 4 (60 mg, 0.19 mmol) was dissolved in 3 mL of acetonitrile, and 41 mg (0.57 mmol) of pyrrolidine and 96 mg (0.96 mmol) of TEA were added dropwise, and the mixture was warmed to reflux overnight. After completion of the reaction, the mixture was evaporated. EtOAcjjjjjjjjj Yield 60%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.33 (d, J = 9.0 Hz, 1H), 7.86 (d, J = 8.0 Hz, 1H), 7.62 (t, J = 7.5 Hz, 1H), 7.55 (t, J = 8.5 Hz, 2H), 7.34 (t, J = 7.5 Hz, 1H), 6.76 (d, J = 8.5 Hz, 2H), 6.38 (s, 1H), 3.91 (t, J=6.0 Hz, 2H), 2.95-2.88 (m, 6H), 2.12-2.07 (m, 2H), 1.98-1.93 (m, 2H); ¹³ C NMR (125MHz, CDCl ₃ ): δ 174.19, 155.44, 146.00 , 136.03, 127.08, 124.08, 122.23, 120.66, 120.41, 118.90, 113.92, 109.83, 102.77, 60.93, 48.61, 47.68, 22.01, 18.61; ESI-MS: m/z = 349 [M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)quinoline-4(1H)-one (I-2).

The preparation was carried out in the same manner as the compound I-1, and the pyrrolidine was replaced with piperidine to give a yellow solid. Yield 75%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.33 (d, J = 8.5 Hz, 1H), 7.79 (d, J = 8.5 Hz, 1H), 7.62 (t, J = 7.5 Hz, 1H), 7.59 (t, J = 8.5 Hz, 2H), 7.34 (t, J = 7.5 Hz, 1H), 6.84 (d, J = 8.5 Hz, 2H), 6.39 (s, 1H), 3.95 (t, J=6.0 Hz, 2H), 2.59-2.54 (m, 6H), 2.04-1.99 (m, 2H), 1.70-1.66 (m, 4H), 1.51-1.48 (m, 2H); ¹³ C NMR (125 MHz, CDCl ₃ ): δ 174.16, 155.86, 145.64, 135.72, 127.17, 123.84, 121.89, 120.82, 120.43, 118.94, 113.68, 110.07, 102.82, 61.52, 50.78, 49.56, 21.23, 20.50, 19.15; ESI-MS: m/z= 363[M+H] ⁺ .

Example 2: Synthesis of I-3 to I-6 in a class I compound (quinolinone).

Step 1: 2-(4-(3-Chloropropoxy)phenyl)-1-methylquinolin-4(1H)-one (Intermediate 5a).

Intermediate 4 (276 mg, 0.88 mmol) was dissolved in 5 mL of DMF, EtOAc (EtOAc, EtOAc (EtOAc) Reaction for 30 min. The reaction solution was poured in 50mL H ₂ O, extracted with EtOAc, washed by water, saturated NaCl, dried over anhydrous Na ₂ SO _4. After recovering the solvent, it was purified by column chromatography (EtOAc:EtOAc:EtOAc Yield 87%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.17 (d, J = 8.0 Hz, 1H), 8.10-8.08 (m, 3H), 7.71 (d, J = 8.0, 1H), 7.48 (t, J = 7.5 Hz, 1H), 7.14 (s, 1H), 7.05 (d, J = 9.0 Hz, 2H), 4.21 (t, J = 6.0 Hz, 2H), 4.12 (s, 3H), 3.79 (t, J = 6.5 Hz, 2H), 2.31-2.26 (m, 2H); ESI-MS: m/z = 328 [M+H] ⁺ .

Chloropropoxy)phenyl)-1-ethylquinolin-4(1H)-one (interm. 5b).

The preparation was carried out in the same manner as the compound intermediate 5a. Yield 67%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.21 (d, J = 8.0 Hz, 1H), 8.09-8.07 (m, 3H), 7.71 (d, J = 8.0, 1H), 7.48 (t, J = 7.5 Hz, 1H), 7.12 (s, 1H), 7.05 (d, J = 9.0 Hz, 2H), 4.37 (q, J = 7.0 Hz, 2H), 4.21 (t, J = 6.0 Hz) , 2H), 3.79 (t, J = 6.5 Hz, 2H), 2.31-2.26 (m, 2H), 1.62 (s, 3H); ESI-MS: m/z = 342 [M+H] ⁺ .

Step 2. 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)-1-methylquinolin-4(1H)-one (I-3).

The preparation method is the same as the compound I-1, the intermediate 5a is used instead of the intermediate 4, and the secondary amine is pyrrolidine to obtain a white solid I-3, the yield is 74%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.17 (d, J = 8.0 Hz, 1H), 8.09 - 8.06 (m, 3H), 7.70 (t, J = 8.0, 1H), 7.47 (t, J = 7.5 Hz, 1H), 7.14 (s, 1H), 7.04 (d, J = 9.0 Hz, 2H), 4.13 (t, J = 6.0 Hz, 2H), 4.12 (s, 3H), 2.71 (t, J = 6.5 Hz, 2H), 2.61-2.57 (m, 4H) ), 2.10-2.05 (m, 2H), 1.85-1.80 (m, 4H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 162.70, 160.18, 158.36, 149.15, 132.72, 129.87, 128.95, 128.79, 125.01, 121.57, 120.15, 114.66, 97.40, 66.51, 55.59, 54.28, 53.17, 28.82, 23.44; ESI-MS: m/z = 363[M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)-1-methylquinolin-4(1H)-one (I-4).

The preparation method is the same as the compound I-1, the intermediate 5a is used instead of the intermediate 4, and the second amine is replaced with piperidine instead of pyrrolidine to obtain a white solid I-4 in a yield of 71%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.17 (d, J = 8.0 Hz, 1H), 8.09-8.06 (m, 3H), 7.70 (d, J = 8.0, 1H), 7.47 (t, J = 7.5 Hz, 1H), 7.14 (s, 1H), 7.04 (d, J = 9.0 Hz, 2H), 4.12 (s, 3H), 4.11 (t, J = 6.0 Hz, 2H), 2.58-2.52 (m, 2H), 2.50-2.44 (m, 4H) ), 2.08-2.03 (m, 2H), 1.66-1.62 (m, 4H), 1.47-1.45 (m, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 162.70, 160.22, 158.41, 149.18, 132.75, 129.92 , 128.99, 128.83, 125.06, 121.61, 120.18, 114.69, 97.46, 66.65, 56.04, 55.64, 54.69, 26.80, 25.97, 24.43; ESI-MS: m/z = 377 [M+H] ⁺ .

Pyrrolidin-1-yl)propoxy)phenyl)-1-ethylquinolin-4(1H)-one (I-5).

The preparation method is the same as the compound I-1, the intermediate 5b is replaced by the intermediate 5b, and the secondary amine is pyrrolidine to obtain a pale yellow solid I-5. Yield 74%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.21 (d, J = 8.0 Hz, 1H), 8.07-8.05 (m, 3H), 7.70 (t, J = 8.0, 1H), 7.47 (t, J = 7.5 Hz, 1H), 7.12 (s, 1H), 7.04 (d, J = 9.0 Hz, 2H), 4.37 (q, J = 7.0 Hz, 2H), 4.13 (t, J = 6.0 Hz) , 2H), 2.77-2.75 (m, 2H), 2.71-2.64 (m, 4H), 2.14-2.10 (m, 2H), 1.89-1.85 (m, 4H), 1.62 (t, J = 7.0 Hz, 3H ¹³ C NMR (100 MHz, CDCl ₃ ): δ 161.98, 160.17, 158.35, 149.25, 132.85, 129.77, 128.96, 128.78, 124.86, 121.69, 120.26, 114.67, 97.95, 66.53, 63.97, 54.27, 53.16, 28.82, 23.46, ESI-MS: m/z = 377 [M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)-1-ethylquinolin-4(1H)-one (I-6).

The preparation method is the same as the compound I-1, the intermediate 5b is replaced by the intermediate 5b, and the secondary amine is piperidine to obtain a pale yellow solid I-6. Yield 85%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.21 (d, J = 8.0 Hz, 1H), 8.07-8.05 (m, 3H), 7.70 (t, J = 8.0, 1H), 7.47 (t, J = 7.5 Hz, 1H), 7.12 (s, 1H), 7.04 (d, J = 9.0 Hz, 2H), 4.37 (q, J = 7.0 Hz, 2H), 4.11 (t, J = 6.0 Hz) , 2H), 2.60-2.52 (m, 2H), 2.51-2.44 (m, 4H), 2.10-2.05 (m, 2H), 1.68-1.63 (m, 4H), 1.62 (t, J = 7.0 Hz, 3H ), 1.50-1.45 (m, 2H); ¹³ C NMR (100 MHz, CDCl ₃ ): δ 162.00, 160.18, 158.40, 149.25, 132.85, 129.83, 128.96, 128.81, 124.91, 121.73, 120.26, 114.68, 98.00, 66.63, 64.01 , 56.00, 54.66, 26.77, 25.93, 24.41, 14.59; ESI-MS: m/z = 391 [M+H] ⁺ .

Example 3: Synthesis of I-7, I-8 in a class I compound (quinolinone).

Step 1: 4-(3-Chloropropoxy)acetophenone (Intermediate 6).

p-Hydroxyacetophenone (5.0 g, 36.7 mmol), 1,3-bromochloropropane (7.3 mL, 73.5 mmol) and K ₂ CO ₃ (10 g, 73.5 mmol) were dissolved in 30 mL of acetonitrile and heated to reflux for 10 h. Excess K ₂ CO _{3 was} removed by suction filtration, and the solvent was evaporated evaporated evaporated. Yield 98%; ¹ H NMR (500MHz, CDCl ₃ ): δ 7.95 (d, J = 9.0 Hz, 2H), 6.95 (d, J = 9.0 Hz, 2H), 4.20 (t, J = 6.0 Hz, 2H), 3.77 (t, J = 6.0 Hz, 2H), 2.56 (s, 3H), 2.29-2.24 (m, 2H); ESI-MS: m/z = 213 [M+H] ⁺ .

Step 2. 2-Bromo-1-(4-(3-chloropropoxy)phenyl)ethanone (Intermediate 7).

Intermediate 6 (2.12g, 10mmol) was dissolved in 20mL diethyl ether was slowly added dropwise at 0 ℃ Br ₂ (0.51mL, 10mmol), dropwise, stirred at room temperature 16h. The reaction solution was poured into saturated NaHCO ₃ solution, extracted with ether, and dried Na ₂ SO ₄ anhydrous organic layer was separated by column chromatography (petroleum _{ether: CH 2 Cl 2 = 3:} 1), to give a pale yellow solid 2.45 g. Intermediate 84%; ¹ H NMR (500MHz, CDCl ₃ ): δ 7.97 (d, J = 9.0 Hz, 2H), 6.97 (d, J = 9.0 Hz, 2H), 4.40 (s, 2H), 4.22 ( t, J = 6.0 Hz, 2H), 3.77 (t, J = 6.0 Hz, 2H), 2.30-2.25 (m, 2H); ESI-MS: m/z = 291 [M+H] ⁺ .

Step 3. 2-(4-(3-Chloropropoxy)phenyl)-2-oxoethyl-2-aminobenzoate (Intermediate 8).

Anthranilic acid (720 mg, 5.25 mmol) and K ₂ CO ₃ (760 mg, 5.5 mmol) were dissolved in 10 mL DMF and stirred at room temperature for 30 min, then intermediate 7 (1.46 g, 5.0 mmol) was added and warmed to 50 ° C After reacting for 3 h, the reaction mixture was poured into EtOAc EtOAc (EtOAc) 1) A white solid of 1.6 g was obtained. Yield 92%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.02 (dd, J = 8.0, 1.0 Hz, 1H), 7.96 (d, J = 8.5 Hz, 2H), 7.32 (t, J = 7.5 Hz, 1H), 6.98 (d, J = 8.5 Hz, 2H), 6.72-6.68 (m, 2H), 5.49 (s, 2H), 4.22 (t, J = 6.0 Hz, 2H), 3.77 (t, J = 6.0 Hz, 2H), 2.30-2.25 (m, 2H); ESI-MS: m/z = 348[M+H] ⁺ .

Step 4. 2-(4-(3-Chloropropoxy)phenyl)-3-hydroxyquinolin-4(1H)-one (Intermediate 9).

Intermediate 8 (1.6 g, 4.6 mmol) and ammonium acetate (5.3 g, 69 mmol) were dissolved in 30 mL of acetic acid and heated to reflux for 3 h. After completion of the reaction, the reaction mixture was poured into 250 mL of water, and a large amount of solid was precipitated, filtered, washed with water to neutral, and dried to give a pale yellow solid (1.02 g). Yield 67%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.32 (d, J = 8.5 Hz, 1H), 7.80-7.76 (m, 2H), 7.61 - 7.57 (m, 2H), 7.32 (t) , J=7.5 Hz, 1H), 6.97-6.95 (m, 2H), 6.72-6.68 (m, 2H), 4.12 (t, J = 6.0 Hz, 2H), 3.76 (t, J = 6.0 Hz, 2H) , 2.26-2.23 (m, 2H); ESI-MS: m/z = 330 [M+H] ⁺ .

Step 5. 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)-3-hydroxyquinolin-4(1H)-one (I-7).

The preparation was carried out in the same manner as the compound I-1, using Intermediate 9 instead of Intermediate 4 to give a beige solid. Yield 59%; ¹ H NMR (500MHz, DMSO-d ₆ ): δ 11.48 (s, 1H), 8.29 (s, 1H), 8.13 (dd, J = 8.0, 1.0 Hz, 1H), 7.78 (d) , J=9.0Hz, 2H), 7.73 (d, J=8.5Hz, 1H), 7.60 (t, J=7.5Hz, 1H), 7.28 (t, J=7.5Hz, 1H), 7.13(d, J = 9.0 Hz, 2H), 4.12 (t, J = 6.5 Hz, 2H), 2.56 (t, J = 7.0 Hz, 2H), 2.46 - 2.42 (m, 4H), 1.95-1.89 (m, 2H), 1.70 -1.67 (m, 4H); ¹³ C NMR (125MHz, DMSO-d ₆ ): δ 170.22, 159.80, 138.42, 138.01, 131.81, 131.11, 130.85, 124.85, 124.80, 122.24, 122.15, 118.84, 114.66, 66.54, 54.13, 52.68, 28.63, 23.58; ESI-MS: m/z = 365 [M+H] ⁺ .

Pyrrolidin-1-yl)propoxy)phenyl)-3-hydroxyquinolin-4(1H)-one (I-8).

The preparation was carried out in the same manner as the compound I-1, and the intermediate 9 was used instead of the intermediate 4, and the piperidine was used instead of the pyrrolidine to obtain a yellow solid. Yield 67%; ¹ H NMR (500MHz, DMSO-d ₆ ): δ 11.47 (s, 1H), 8.28 (s, 1H), 8.12 (dd, J = 8.0, 1.0 Hz, 1H), 7.77 (d) , J=9.0Hz, 2H), 7.72 (d, J=8.5Hz, 1H), 7.59 (t, J=7.5Hz, 1H), 7.27(t, J=7.5Hz, 1H), 7.12(d, J = 9.0 Hz, 2H), 4.12 (t, J = 6.5 Hz, 2H), 2.56 (t, J = 7.0 Hz, 2H), 2.46 - 2.42 (m, 4H), 1.95-1.89 (m, 2H), 1.70 -1.67 (m, 4H), 1.51-1.48 (m, 2H); ¹³ C NMR (125MHz, DMSO-d ₆ ): δ170.25, 159.49, 138.43, 138.03, 131.75, 131.17, 130.84, 125.04, 124.83, 122.26, 122.14 , 118.88, 114.67, 65.99, 54.29, 53.08, 28.58, 24.19, 23.74; ESI-MS: m/z = 379 [M+H] ⁺ .

Example 4: Synthesis of I-9, I-10 in a class I compound (quinolinone).

Step 1. 2-(4-(3-Chloropropoxy)phenyl)-3-methoxy-1-methylquinolin-4(1H)-one (Intermediate 10).

Intermediate 9 (660 mg, 2.0 mmol), dimethyl sulfate (0.75 mL, 8.0 mmol) and K ₂ CO ₃ (1.1 g, 8.0 mmol) were dissolved in 10 mL of acetone and heated to reflux for 4 h. After completion of the reaction, the mixture was filtered under suction and filtered. Yield 78%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.59 (dd, J = 8.0, 1.0 Hz, 1H), 7.71 (t, J = 7.5 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.42 (t, J = 7.5 Hz, 1H), 7.31 (d, J = 8.5 Hz, 2H), 7.07 (d, J = 8.5 Hz, 2H), 4.22 (t, J = 6.0 Hz, 2H), 3.81 (t, J = 6.0 Hz, 2H), 3.65 (s, 3H), 3.54 (s, 3H), 2.36-2.24 (m, 2H); ESI-MS: m/z = 358 [M+ H] ⁺ .

Step 2, 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)-3-methoxy-1-methylquinolin-4(1H)-one (I-9 ).

The preparation was carried out in the same manner as Compound I-1, using Intermediate 10 instead of Intermediate 4. Yield: 65%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.59 (dd, J = 8.0, 1.5 Hz, 1H), 7.70 (t, J = 8.0 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.40 (t, J = 7.5 Hz, 1H), 7.28 (d, J = 8.0 Hz, 2H), 7.05 (d, J = 8.5 Hz, 2H), 4.12 (t, J = 6.0 Hz) , 2H), 3.65 (s, 3H), 3.53 (s, 3H), 2.70 (t, J = 7.5 Hz, 2H), 2.61-2.55 (m, 4H), 2.13 - 2.05 (m, 2H), 1.85- 1.81 (m, 4H); ¹³ C NMR (125MHz, CDCl ₃ ): δ 172.96, 159.69, 147.31, 141.26, 140.20, 131.85, 130.38, 127.14, 126.79, 124.35, 122.99, 115.78, 114.67, 66.46, 59.89, 54.30, 53.15 , 37.12, 28.74, 23.47; ESI-MS: m/z = 393 [M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)-3-methoxy-1-methylquinolin-4(1H)-one (I-10).

The preparation was carried out in the same manner as the compound I-1, using the intermediate 10 instead of the intermediate 4, and the piperidine was used in place of the pyrrolidine to give an off-white solid. Yield: 67%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.58 (d, J = 8.0 Hz, 1H), 7.70 (t, J = 7.5 Hz, 1H), 7.53 (d, J = 8.5 Hz) , 1H), 7.41 (t, J = 7.5 Hz, 1H), 7.29 (d, J = 8.5 Hz, 2H), 7.05 (d, J = 8.5 Hz, 2H), 4.10 (t, J = 6.0 Hz, 2H) ), 3.65 (s, 3H), 3.53 (s, 3H), 2.57-2.51 (m, 2H), 2.47-2.43 (m, 4H), 2.08-2.03 (m, 2H), 1.66-1.58 (m, 4H) ), 1.48-1.44 (s, 2H); ¹³ C NMR (125 MHz, CDCl ₃ ): δ 172.95, 159.72, 147.28, 141.27, 140.21, 131.82, 130.38, 127.15, 126.79, 124.36, 122.97, 115.77, 114.68, 66.60, 59.88 , 55.96, 54.66, 37.10, 26.77, 25.92, 24.39; ESI-MS: m/z = 407 [M+H] ⁺ .

Example 5: Synthesis of II-1, II-2 in a class II compound (flavonoids).

Step 1: 4-(3-Chloropropoxy)benzaldehyde (Intermediate 11).

The preparation method was the same as that of the intermediate 6, and p-hydroxybenzaldehyde was used instead of p-hydroxyacetophenone to give a pale yellow solid. Yield: 98%; ¹ H NMR (500MHz, CDCl ₃ ): δ 9.89 (s, 1H), 7.85 (d, J = 9.0 Hz, 2H), 7.02 (d, J = 9.0 Hz, 2H), 4.22 (t, J = 6.0 Hz, 2H), 3.77 (t, J = 6.0 Hz, 2H), 2.30-2.25 (m, 2H); ESI-MS: m/z = 199 [M+H] ⁺ .

Step 2: 1-(2-Hydroxyphenyl)-3-(4-(3-chloropropoxy)phenyl)prop-2-en-1-one (Intermediate 12).

p-Hydroxyacetophenone (1.36 g, 10 mmol), Intermediate 11 (1.98 g, 10 mmol) was dissolved in 15 mL of C ₂ H ₅ OH, and 1.68 g KOH (30 mmol) was added and refluxed for 2 h. After cooling, a part of the solvent was removed under reduced pressure, and ethyl acetate (200 mL) was added to the residue, and the mixture was adjusted to pH 4-5 with 2N HCl. Yield 89%; ¹ H NMR (500MHz, CDCl ₃ ): δ 12.92 (s, 1H), 7.94-7.89 (m, 2H), 7.63 (d, J = 8.5 Hz, 2H), 7.57 and 7.54 (s) , 1H), 7.49 (t, J = 7.5 Hz, 1H), 7.03 (d, J = 8.5 Hz, 1H), 6.97 - 6.93 (m, 3H), 4.20 (t, J = 6.0 Hz, 2H), 3.78 (t, J = 6.5 Hz, 2H), 2.30-2.25 (m, 2H); ESI-MS: m/z = 317 [M+H] ⁺ .

Step three: 2-(4-(3-Chloropropoxy)phenyl)-3-hydroxy-4H-benzopyran-4-one (Intermediate 13).

Intermediate 12 (1.05 g, 3 mmol) was dissolved in 10 mL of C ₂ H ₅ OH, 10 mL of 0.5N KOH solution was added, and then 0.6 mL of H ₂ O ₂ aqueous solution (30%) was added in portions, and stirred at room temperature for 1 h. The reaction solution was poured into ice water, a large amount of solid was precipitated, and filtered, and the filter cake was washed with water and dried to give 930 mg of a yellow solid. Yield 94%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.61 (d, J = 7.5 Hz, 1H), 8.04 (d, J = 9.0 Hz, 2H), 7.61 - 7.58 (m, 2H), 7.28 (t, J = 7.0 Hz, 1H), 7.01 (d, J = 9.0 Hz, 2H), 4.15 (t, J = 6.0 Hz, 2H), 3.83 (t, J = 6.5 Hz, 2H), 2.23 2.17 (m, 2H); ESI-MS: m/z = 331 [M+H] ⁺ .

Step 4: 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)-3-hydroxy-4H-benzopyran-4-one (II-1).

The preparation was carried out in the same manner as Compound I-1, and Intermediate 13 was used instead of Intermediate 4 to give a yellow solid. ^{Yield: 68%; 1 H NMR (} 500MHz, CDCl 3): δ8.25-8.21 (m, 3H), 7.71 (t, J = 7.5Hz, 1H), 7.59 (d, J = 8.5Hz, 1H) , 7.42 (t, J = 7.5 Hz, 1H), 7.06 (d, J = 9.0 Hz, 2H), 4.14 (t, J = 6.0 Hz, 2H), 2.74 - 2.67 (m, 2H), 2.64 - 2.58 ( m, 4H), 2.11-2.06 (m, 2H), 1.86-1.80 (m, 4H); ¹³ C NMR (125MHz, DMSO-d ₆ ): δ 173.10, 160.32, 154.88, 146.04, 138.65, 133.96, 129.87, 125.20 , 124.95, 123.92, 121.81, 118.80, 114.94, 66.52, 54.10, 52.64, 28.56, 23.56; ESI-MS: m/z = 366 [M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)-3-hydroxy-4H-benzopyran-4-one (II-2).

The preparation was carried out in the same manner as the compound I-1, the intermediate 13 was replaced with the intermediate 13 and the piperidine was used instead of the pyrrolidine to give a yellow solid. Yield: 61%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.25 - 8.21 (m, 3H), 7.71 (t, J = 7.5 Hz, 1H), 7.59 (d, J = 8.5 Hz, 1H) , 7.42 (t, J = 7.5 Hz, 1H), 7.06 (d, J = 9.0 Hz, 2H), 4.12 (t, J = 6.5 Hz, 2H), 2.56 (t, J = 7.0 Hz, 2H), 2.46 -2.42 (m, 4H), 1.95-1.89 (m, 2H), 1.70-1.67 (m, 4H), 1.52-1.48 (m, 2H); ¹³ C NMR (125MHz, DMSO-d ₆ ): δ 173.47, 160.24 , 154.85, 145.98, 138.02, 133.85, 129.77, 125.19, 124.87, 124.09, 121.78, 118.78, 114.93, 66.61, 55.56, 54.59, 26.69, 26.07, 24.61; ESI-MS: m/z = 380 [M+H] ⁺ .

Example 6: Synthesis of II-3, II-4 in a class II compound (flavonoids).

Step 1: 2-(4-(3-Chloropropoxy)phenyl)-3-methoxy-4H-benzopyran-4-one (Intermediate 14).

Intermediate 13 (660 mg, 2.0 mmol), iodomethane (8.0 mmol) and K ₂ CO ₃ (1.1 g, 8.0 mmol) were dissolved in 10 mL of acetone and heated to reflux for 4 h. After completion of the reaction, the mixture was filtered under suction and filtered. Yield 78%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.59 (dd, J = 8.0, 1.0 Hz, 1H), 7.71 (t, J = 7.5 Hz, 1H), 7.53 (d, J = 8.5 Hz, 1H), 7.42 (t, J = 7.5 Hz, 1H), 7.31 (d, J = 8.5 Hz, 2H), 7.07 (d, J = 8.5 Hz, 2H), 4.22 (t, J = 6.0 Hz, 2H), 3.81 (t, J = 6.0 Hz, 2H), 3.65 (s, 3H), 3.54 (s, 3H), 2.36-2.24 (m, 2H); ESI-MS: m/z = 358 [M+ H] ⁺ .

Step 2. 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)-3-methoxy-4H-benzopyran-4-one (II-3).

The preparation was carried out in the same manner as Compound I-1, and Intermediate 14 was used instead of Intermediate 4 to give a red solid. Yield: 54%; ¹ H NMR (500MHz, CDCl ₃ ): δ 8.26 (d, J = 8.0 Hz, 1H), 8.12 (d, J = 8.5 Hz, 2H), 7.68-7.64 (m, 1H) , 7.53-7.50 (m, 1H), 7.40-7.37 (m, 1H), 7.00 (d, J = 8.5 Hz, 2H), 4.12 (t, J = 6.0 Hz, 2H), 3.90 (s, 3H), 2.74-2.67 (m, 2H), 2.64-2.58 (m, 4H), 2.11-2.06 (m, 2H), 1.86-1.80 (m, 4H); ¹³ C NMR (125MHz, CDCl ₃ ): δ 175.01, 161.01, 155.72, 155.15, 140.80, 133.28, 130.24, 125.77, 124.58, 124.20, 123.07, 117.90, 114.50, 66.52, 59.92, 54.29, 53.07, 28.68, 23.46; ESI-MS: m/z = 380 [M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)-3-methoxy-4H-benzopyran-4-one (II-4).

The preparation was carried out in the same manner as the compound I-1, and the intermediate 14 was used instead of the intermediate 4, and the piperidine was used instead of the pyrrolidine to obtain a yellow solid. Yield: 57%; ¹ H NMR (500MHz, CDCl ₃ ): 8.26 (d, J = 8.0 Hz, 1H), 8.12 (d, J = 8.5 Hz, 2H), 7.68-7.64 (m, 1H), 7.53 -7.50 (m, 1H), 7.40-7.37 (m, 1H), 7.00 (d, J = 8.5 Hz, 2H), 4.12 (t, J = 6.5 Hz, 2H), 3.89 (s, 3H), 2.56 ( t, J=7.0 Hz, 2H), 2.46-2.42 (m, 4H), 1.95-1.89 (m, 2H), 1.70-1.67 (m, 4H), 1.52-1.48 (m, 2H); ¹³ C NMR ( 125 MHz, CDCl ₃ ): δ 175.00, 161.03, 155.71, 155.16, 140.82, 133.27, 130.24, 125.80, 124.57, 124.22, 123.09, 117.90, 114.51, 66.65, 59.93, 55.89, 54.67, 26.68, 25.90, 24.38; ESI-MS: m/z = 394 [M+H] ⁺ .

Example 7: Synthesis of II-5, II-6 in a class II compound (flavonoids).

Step 1: 1-(2,6-Dihydroxyphenyl)-3-(4-(3-chloropropoxy)phenyl)prop-2-en-1-one (Intermediate 15).

The preparation was carried out in the same manner as the intermediate 12, using 2,6-dihydroxyacetophenone in place of o-hydroxyacetophenone to give a yellow solid. Yield: 85%; ESI-MS: m/z = 333 [M+H] ⁺ .

Step 2: 2-(4-(3-Chloropropoxy)phenyl)-5-hydroxy-4H-benzopyran-4-one (Intermediate 16)

Intermediate 15 (1.00g, 3.2mmol) was dissolved in 15mL DMSO was added I ₂ (128mg, 0.5mmol) and 0.5mL of concentrated sulfuric acid was stirred at 85 ℃ 24h. The reaction solution was poured into 200mL H ₂ O, extracted three times with EtOAc, the combined organic layers were washed by water, saturated NaCl, dried over anhydrous Na ₂ SO _4, then separated by column chromatography (petroleum ether: EtOAc = 4: 1 ), 565 mg of a white solid. Yield: 45%; ¹ H NMR (500MHz, CDCl ₃ ): δ 7.88 (d, J = 9.0 Hz, 2H), 7.55 (t, J = 8.5 Hz, 1H), 7.05 (d, J = 9.0 Hz) , 2H), 7.00 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 6.66 (s, 1H), 4.23 (t, J = 6.0 Hz, 2H), 3.79 (t , J=6.0 Hz, 2H), 2.32-2.27 (m, 2H); ESI-MS: m/z = 331 [M+H] ⁺ .

Step 3: 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)-5-hydroxy-4H-benzopyran-4-one (II-5).

The preparation was carried out in the same manner as Compound I-1, and Intermediate 16 was used instead of Intermediate 4 to give a yellow solid. Yield: 58%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 7.88 (d, J = 9.0 Hz, 2H), 7.55 (t, J = 8.5 Hz, 1H), 7.05 (d, J = 9.0 Hz) , 2H), 7.00 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 6.66 (s, 1H), 4.12 (t, J = 6.0 Hz, 2H), 2.78 (t , J=6.0 Hz, 2H), 2.65-2.61 (m, 4H), 2.11-2.06 (m, 2H), 1.89-1.86 (m, 4H); ESI-MS: m/z = 366 [M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)-5-hydroxy-4H-benzopyran-4-one (II-6).

The preparation was carried out in the same manner as the compound I-1, and the intermediate 16 was used instead of the intermediate 4, and the piperidine was used instead of the pyrrolidine to obtain a yellow solid. Yield: 64%; ¹ H NMR (500MHz, CDCl ₃ ): δ 7.87 (d, J = 9.0 Hz, 2H), 7.56 (t, J = 8.5 Hz, 1H), 7.04 (d, J = 9.0 Hz) , 2H), 6.99 (d, J = 8.0 Hz, 1H), 6.81 (d, J = 8.0 Hz, 1H), 6.65 (s, 1H), 4.13 (t, J = 6.0 Hz, 2H), 2.59-2.53 (m, 2H), 2.50-2.44 (m, 4H), 2.10-2.04 (m, 2H), 1.70-1.62 (m, 4H), 1.48-1.45 (m, 2H); ESI-MS: m/z = 380 [M+H] ⁺ .

Example 8: Synthesis of II-7, II-8 in a class II compound (flavonoids).

Step 1: 2-(4-(3-Chloropropoxy)phenyl)-5-methoxy-4H-benzopyran-4-one (Intermediate 17).

The preparation was carried out in the same manner as the intermediate 14 and Intermediate 16 was used instead of Intermediate 13. Yield: 75%; ¹ H NMR (500MHz, CDCl ₃ ): δ 7.88 (d, J = 9.0 Hz, 2H), 7.55 (t, J = 8.5 Hz, 1H), 7.05 (d, J = 9.0 Hz) , 2H), 6.99 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 6.68 (s, 1H), 4.21 (t, J = 6.0 Hz, 2H), 3.90 (s , 3H), 3.78 (t, J = 6.0 Hz, 2H), 2.32 - 2.27 (m, 2H); ESI-MS: m/z = 345 [M+H] ⁺ .

Step 2: 2-(4-(3-(Pyrrolidin-1-yl)propoxy)phenyl)-5-methoxy-4H-benzopyran-4-one (II-7).

The preparation was carried out in the same manner as the compound I-1, using Intermediate 17 instead of Intermediate 4 to give a yellow solid. Yield: 61%; ¹ H NMR (500MHz, CDCl ₃ ): δ 7.87 (d, J = 9.0 Hz, 2H), 7.55 (t, J = 8.5 Hz, 1H), 7.05 (d, J = 9.0 Hz) , 2H), 6.99 (d, J = 8.0 Hz, 1H), 6.82 (d, J = 8.0 Hz, 1H), 6.68 (s, 1H), 4.13 (t, J = 6.0 Hz, 2H), 3.91 (s , 3H), 2.76 (t, J = 6.0 Hz, 2H), 2.65-2.60 (m, 4H), 2.10-2.05 (m, 2H), 1.89-1.85 (m, 4H); ESI-MS: m/z =380[M+H] ⁺ .

Piperidin-1-yl)propoxy)phenyl)-5-methoxy-4H-benzopyran-4-one (II-8).

The preparation was carried out in the same manner as the compound I-1, using the intermediate 17 instead of the intermediate 4, and the piperidine was used in place of pyrrolidine to give a yellow solid. Yield: 67%; ¹ H NMR (500MHz, CDCl ₃ ): δ 7.88 (d, J = 9.0 Hz, 2H), 7.56 (t, J = 8.5 Hz, 1H), 7.05 (d, J = 9.0 Hz) , 2H), 7.00 (d, J = 8.0 Hz, 1H), 6.83 (d, J = 8.0 Hz, 1H), 6.66 (s, 1H), 4.12 (t, J = 6.0 Hz, 2H), 3.89 (s , 3H), 2.54-2.49 (m, 2H), 2.48-2.41 (m, 4H), 2.06-2.00 (m, 2H), 1.65-1.59 (m, 4H), 1.47-1.43 (m, 2H); - MS: m/z = 394 [M+H] ⁺ .

Example 9: Histamine H ₃ receptor antagonistic activity of phenylquinolinones and flavonoid derivatives.

This section uses Clobenpropit as a positive control, and screens the histamine H ₃ receptor antagonistic activity of the 18 target compounds synthesized by LANCE Ultra TR-FRET method and ThT fluorescence assay, respectively. Other compounds of the present invention are listed below. The compounds have similar beneficial effects, but it should not be understood that the compounds of the invention have only the following beneficial effects. The results are shown in Table 3.

Table 3 Histamine H ₃ receptor antagonistic activity of phenylquinolinone and flavonoid derivatives

It can be seen from the receptor antagonistic activity in Table 3 that the selected phenylquinolinone compounds exhibit strong histamine H ₃ receptor inhibitory activity, and most of the compounds have activity comparable to Clobenpropit, and have six molecular activities more than Clobenpropit. good.

Example 10 Determination of anti-amyloid peptide aggregation activity.

A ThT fluorescence test.

(1) Solution and sample preparation: Prepared in PBS buffer (10 mM, pH 7.4): Weigh 57.996 g of Na ₂ HPO ₄ ·12H ₂ O, 5.928 g of NaH ₂ PO ₄ ·2H ₂ O dissolved in 1000 mL of water to prepare a concentration. It is a 0.2M PBS stock solution; take 50mL of stock solution, dilute to 1000mL with water, and filter by 0.22μm microporous membrane. Gly-NaOH buffer preparation (50 mM, pH 8.5): Weigh 3.7535 g of glycine dissolved in 800 mL of water, slowly add 0.1 M NaOH solution to adjust the pH to 8.5, and bring up to 1000 mL with water. Preparation of Aβ monomer: 1 mg of Aβ _{1-42 was} dissolved in 1 mL of HFIP, sonicated for 10 min, dispensed, and allowed to stand at room temperature for 12 h, vacuum dried, and stored in a -20 ° C refrigerator. The Aβ monomer was reconstituted in DMSO to 2 mM, and after briefly vortexing, it was formulated into a 50 μM stock solution in 10 mM PBS buffer. Compound Formulation: The compound was dissolved in DMSO to prepare a stock solution at a concentration of 10 mM. Dilute to the corresponding concentration with PBS. Preparation of ThT solution (5 μM): 1.59 g of ThT was weighed and dissolved in 100 mL of Gly-NaOH buffer to prepare a stock solution having a concentration of 50 mM, and stored in the dark. 10 μL of the stock solution was diluted to 100 mL with Gly-NaOH buffer to obtain a 5 μM ThT solution.

(2) Aβ _1-42 and a compound (positive control or PBS buffer) were added to the EP tube to a final concentration of 25 μM and 20 μM, respectively, and placed at 37 ° C under constant temperature (150 rpm) for 24 hours. Add 20 μL of the incubation solution to a 96-well plate, add 180 μL of 5 μM ThT solution, incubate for 5 min in the dark, and use a multi-function microplate reader to measure the fluorescence intensity at 485 nm of 450 nm excitation.

The fluorescence value after incubation with Aβ _1-42 alone was used as a control. In order to avoid the interference of the fluorescence of the compound itself, the fluorescence value of the compound measured by ThT was subtracted as the background.

Table 4 Aβ aggregation inhibitory activity of phenylquinolinone and flavonoid derivatives

From the above experimental results, most of the target molecules designed and synthesized showed good Aβ _1-42 self-aggregation activity, while the positive control Clobenpropit had no such activity.

Compounds I-6 and II-1 were further selected to observe their inhibition of Aβ _1-42 self-aggregation and the activity of depolymerization of the aggregated Aβ _1-42 sample by transmission electron microscopy.

Tested by transmission electron microscopy (TEM).

(1) Solution and sample preparation: HEPES buffer preparation: (20 μM HEPES, pH 6.6, 150 μM NaCl) 47.77 mg HEPES and 87.66 mg NaCl were weighed and dissolved in 100 mL of water to prepare a mother liquor. Dilute 5 mL of mother liquor to 400 mL, slowly add 1 mM NaOH solution to adjust the pH to 6.6, and dilute to 500 mL.

Self-aggregated samples: Aβ _1-42 and compounds (positive control Curcumin or PBS buffer) were added to the EP tube to a final concentration of 25 μM and 20 μM, respectively, and placed at 37 ° C with constant temperature shaking (150 rpm), and incubated for 24 hours.

Deagglomeration of the sample after self-aggregation: Aβ _1-42 (final concentration 50 μM) was added to the EP tube, and after incubation at 37 ° C for 24 h, the compound (positive control Curcumin or PBS buffer) was added to make it with Aβ _1- The final concentrations of ₄₂ were 20 μM and 25 μM, respectively, and incubated for a further 24 h.

(2) 10 μL of the incubation solution was placed on a carbon support membrane copper mesh, allowed to stand for 2 min, and then blotted from the side with filter paper. After standing for 30 s, 5 μL of 1% uranyl acetate solution was added to negatively stain the sample, and after standing for 2 min, Use a filter paper to blot dry from the side, dry it, place it under a transmission electron microscope, and take a photo. Acceleration voltage is 80kV, magnification is 50,000 times

Figure 3 Effect of compound on Aβ self-aggregation, the ratio is 0.5μm

([Aβ _1-42 ] = 25 μM; [compd.] = 20 μM; a. Aβ _1-42 alone, 0h; baAβ _1-42 alone, 24h; c. Aβ _1-42 +I-6, 24h; d. Aβ _1-42 +II-1, 24 h. PBS pH 7.4).

Compounds I-6 and II-1 were subjected to Aβ _1-42 aggregation inhibition experiments. As shown in the above figure, Aβ monomers were able to undergo significant self-aggregation after 24 hours compared to 0 hours (Fig. 3b). The compound was able to significantly inhibit A[beta] self-aggregation (Fig. 3c, 3d).

Further depolymerization experiments are shown in the figure below. The depolymerization effect of the sample added with compound I-6, II-1 is obvious, and the aggregated filaments can be degraded into short rods or granules (Fig. 4b, 4c), and their activity is better than Curcumin (Figure 4d)

Figure 4 Effect of compound on Aβ depolymerization, the ratio is 0.5μm

([Aβ _1-42 ]=25 μM, [compd.]=20 μM; a.Aβ _1-42 alone, 24h; b.Aβ _1-42 fibrils+I-6,24h; c.Aβ _1-42 fibrils+II -1, 24h; d. Aβ _1-42 fibrils + Curcumin, 24 h. PBS pH 7.4, 50,000 x).

Example 11 Effect on ischemic brain injury.

The rats were anesthetized with 10% chloral hydrate (3.5 ml/kg, intraperitoneal injection), fixed on the back, skin incision along the left neck, ligation of the external carotid artery and lateral common carotid artery, and clamping of the telecentric neck with an artery clamp The artery was incision at the bifurcation of the external carotid artery and the internal carotid artery. A tying wire (diameter 0.25 mm, length 18 mm) was inserted, the incision was ligated, the skin was sutured, and the distal carotid artery artery clip was released. After 2 hours, gently lift the remaining thread to a slight resistance to achieve reperfusion of the middle cerebral artery and complete the modeling. In the sham operation group, the rats were not used for the middle artery insertion after anesthesia, and the rest of the operations were consistent. During the experiment, the body temperature was maintained at 37±0.5°C with a constant temperature blanket, and the wound was added with penicillin sodium after the operation. Rats that survived for 24 hours were randomly divided into a sham operation group, a model group, and each test group, and the test group was administered the compounds described in the present invention, and grouped by the number corresponding to the compound.

Each group was intraperitoneally administered, and the rats in the model group and the sham operation group were given an equal volume of physiological saline. After the first administration for 16 hours, each group was intraperitoneally administered once again.

Nerve injury symptom scoring method, 0 points: normal behavior, the two forelimbs symmetrically extended to the ground when lifting; 1 point: the contralateral forearm wrist bending when lifting the tail; 2 points: the contralateral forelimb elbow bending when lifting the tail; 3 Points: The shoulders are rotated while lifting the tail; when walking, they are inclined to the opposite side of the injury; 4 points: no voluntary activity.

The rats were scored for neurological damage at the time of the first administration, 4 hours after the administration, and 20 hours after the administration, and 24 hours after the first administration, the whole brain was quickly taken out after the neck was sacrificed. On the ice, 3 isolated hippocampus were randomly selected from each group to detect histamine content, and the remaining part was frozen at -20 °C. The brain was evenly cut into 5 pieces with a coronal plane, and the brain slices were placed at 1% TTC at room temperature in the dark. Dye for 30 minutes and fix with 4% paraformaldehyde for 30 minutes. Take a photo analysis and calculate the cerebral infarction volume ratio (infarct volume / whole brain volume * 100%)

Table 5 results of nerve injury symptoms, hippocampus histamine content, cerebral infarction volume ratio

Compared with the model group, each test group *P<0.05; #P<0.01; ΔP<0.05; ⊕P<0.01.

The rat behavioral grading method was scored, and the symptoms of nerve injury 4 hours after the first administration, the compound of the present invention was significantly different from the model group, and the compound and model of the present invention were 20 hours after the first administration. The group has significant difference, indicating that the compound of the present invention has a significant improvement on the neurological function of rats with ischemic brain injury; the histamine content in the hippocampus, the compound of the present invention is different from the model group, indicating the present invention The compound can effectively increase the content of histamine in the brain of rats with ischemic brain injury; the infarct volume ratio, the compound of the present invention is significantly different from the model group, indicating that the compound of the present invention can effectively reduce ischemic brain damage The cerebral infarct volume of the mouse.

Example 12, Effect on pentylenetetrazol-induced acute epilepsy in mice

Each group of mice were given the test drug of the present invention and the positive drug phenytoin, and administered intragastrically according to a gavage volume of 0.1 ml/10 g. The model group was given the same volume of physiological saline, and the positive control group was given a dose of phenytoin 50 mg/kg. Each dose of the reagent group was 50 mg/kg, and the compound of the present invention was administered to the reagent group, and the compounds were grouped by the corresponding numbers.

After 1 hour of intragastric administration, intraperitoneal injection of pentylenetetrazol 70 mg/kg began to record myoclonic latency (LTMJ). If there was no seizure within 30 min, the above latency was recorded as 30 min.

The seizure standard is based on the Racine standard, and the aging of the facial muscles or the corners of the mouth, the convulsions of one side of the limbs, the rigidity or the limb twitching epilepsy

Table 6 Results of mouse seizure latency (n=6)

The results showed that compared with the model group, the compound of the present invention can significantly delay acute seizures in mice, and the incubation period is extended by 3 to 5 times; the incubation period of the compound of the present invention is longer than that of the control group of phenytoin. The compounds of the invention have significant anti-epileptic effects.

Claims (6)

1. A benzoquinolinone, a flavonoid derivative, and a pharmaceutically acceptable salt or solvate thereof, having the following general structure (A):

   

   General structure (A)

   In the formula:

   X is selected from N or O;

   R _{1 is} selected from H, C _1-3 alkyl or C _1-3 alkoxy;

   R _{2 is} selected from H, hydroxy or C _1-3 alkoxy;

   R _{3 is} selected from H, hydroxy, C _1-3 alkyl or C _1-3 alkoxy;

   NR'R" is selected from cyclic amines and open-chain alkylamines having a total of 3 to 6 carbon atoms, including but not limited to the following fragments:

   
2. The benzoquinolinone thiazine or flavonoid derivative according to claim 1, which is characterized in that:

   Said

   

   Selected from the following two heterocycles:

   

   R _{1 is} selected from the group consisting of hydrogen, methyl and ethyl; R _{2 is} selected from the group consisting of hydrogen, hydroxy and methoxy; and R _{3 is} selected from the group consisting of hydrogen, hydroxy and methoxy.
3. The benzoquinolinone thiazine or flavonoid derivative according to claim 1, wherein the compound is selected from the group consisting of:

   

   

   And a compound, an acceptable salt, and a solvate, prodrug (ester or phosphate), stereoisomer, or deuterated compound of any one of the compounds.
4. A pharmaceutical composition comprising at least one active ingredient and at least one pharmaceutically acceptable carrier or excipient, said active ingredient being selected from claims 1-3 Any one or more of the compound, the pharmaceutically acceptable salt, and the solvate of the compound.
5. A compound according to any one of claims 1 to 3, an acceptable salt, and a solvate, prodrug (ester or phosphate), stereoisomer, deuterated product of the compound, and in combination with other drugs, The use of a medicament for the treatment of a neurodegenerative disease and an ischemic brain injury.
6. The use according to claim 5, wherein the neurodegenerative disease is selected from the group consisting of Alzheimer's disease, Parkinson's disease, narcolepsy, epilepsy, and gradual freezing.

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