CN109111400B - Preparation and application of phenylquinolinone and flavonoid derivatives - Google Patents

Abstract

The invention relates to phenyl quinolinone and flavonoid derivatives and a preparation method thereof, wherein the derivatives are histamine H₃The receptor antagonist can protect and restore the normal function of the central nervous system, and has great clinical application value, especially in neurodegenerative diseases and ischemic brain injury.

Description

Preparation and Application of Phenylquinolinone and Flavonoid Derivatives

technical field

The invention belongs to the technical field of medicine, and in particular relates to the preparation of phenylquinolinone and flavonoid derivatives. The active ingredient is a histamine H3 receptor antagonist, which can protect and restore the normal function of the central nervous system and prevent neurodegeneration. Application in the treatment of sexual diseases, ischemic ischemic encephalopathy, Parkinson's syndrome, narcolepsy, epilepsy and ALS. The phenylquinolinone and flavonoid derivatives also include their pharmaceutically acceptable salts, complexes, solvates and the like.

Background technique

Histamine _H3 receptors exist in the cerebral cortex, striatum, hippocampus, olfactory bulb, substriatal nucleus, thalamus, lower brainstem spinal nucleus, cerebellum, etc. Histamine H3 receptors are the self _- receptors of neuronal synapses. In vivo, excitation of the receptor inhibits the release of histamine or other neurotransmitters, and antagonism of the receptor promotes the release of neurotransmitters.

Cerebral ischemia is mainly caused by cerebral infarction caused by insufficient cerebral blood supply due to thrombosis, embolism or other reasons in cerebral blood vessels. It is also one of the leading killers of human disability and death. Possibly due to the complex mechanism of the pathological process, so far, no effective drugs have been used in clinical practice.

Studies have shown that histamine _H3 receptor antagonists can promote the synthesis and release of neurotransmitters such as histamine for the treatment of Alzheimer's disease (AD), Parkinson's disease (PD), ALS ( ALS), narcolepsy, etc.

Among them, Alzheimer's disease (AD) has been discovered for more than 100 years, and the number of cases is increasing year by year. Studies have shown that β-amyloid peptide (β-amyloid, Aβ) aggregation, tau protein hyperphosphorylation, mitochondrial dysfunction, oxidation Many factors, such as stress and neurotransmitter system dysfunction, may be related to its pathogenesis, and different molecular mechanisms are related to each other and influence each other. At present, the first-line treatment drugs for AD are mainly acetylcholinesterase (AChE) inhibitor donepezil, rivarastamine, galantamine and non-competitive NMDA receptor antagonist memantine. Although the cognitive-related neurotransmitter system can alleviate the cognitive impairment of early-stage patients, it cannot fundamentally improve the disease state or terminate the disease process.

In view of the fact that the occurrence and development of AD involves a variety of factors, drugs that can simultaneously target multiple targets (multitarget-directed ligands, MTDLs) have unique advantages and have attracted widespread attention. Studies have shown that _H3 receptors are important receptors for neurotransmitter regulation in the brain. Blocking _H3 receptors with _H3 receptor antagonists can effectively promote histamine, acetylcholine and other neurotransmitters related to cognition. Release, at present, a number of histamine H3 receptor antagonists (ABT _- 288, AZD5213, CEP-26401, GSK239512 and MK0249) have entered clinical research for Alzheimer's disease, Parkinson's disease, narcolepsy, etc. The treatment of neurodegenerative diseases is shown in Figure 1.

Senile plaques (senileplaques, SP) formed by the accumulation of amyloid peptide Aβ in the limbic and cerebral cortex are one of the most typical pathological features of AD. Moreover, the oligomers, fibrils and fibers formed during the aggregation process of Aβ can be toxic to nerve cells and even lead to nerve cell apoptosis. Therefore, Aβ aggregation is generally considered to be the intrinsic cause of AD, and drugs that inhibit Aβ aggregation have become an important direction for current AD drug development.

Based on the systematic analysis of the biological characteristics of AD-related targets, the present invention organically combines a symptom-improving target (H ₃ receptor) and a cause-specific target (Aβ aggregation), and uses a rational drug design method. , designed multi-target compounds that can antagonize H ₃ receptors and inhibit the aggregation of Aβ at the same time. Such multi-target compounds can not only promote the release of neurotransmitters, improve the symptoms of AD patients, and delay the disease; Self-aggregation can fundamentally prevent the progression of AD, and achieve the effect of "multi-pronged approach, treating both the symptoms and root causes", which may have a unique effect on the treatment of neurodegenerative diseases such as AD.

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

The molecular structure of histamine _H3 receptor antagonists is mostly chain molecules, mainly composed of three parts: the western part is the nitrogen-containing basic region of aliphatic tertiary amine and the connecting chain part; Fragments are lipid-soluble aromatic rings or basic groups (Figure 2)

In the present invention, starting from the pharmacophore model of _H3 receptor antagonists, benzoquinolinone (benzopyrone) is used as the fat-soluble aromatic ring (eastern fragment), and phenoxypropylamine is used as the other part (central ring). + Western fragment), designed and synthesized new phenylquinolinone and flavonoid derivatives; on the other hand, previous studies showed that N-aminopropoxyphenyl-pyridin-4-one derivatives with similar structures It has good Aβ aggregation inhibitory activity (N-substituted phenylpyridin-4-one derivatives and their preparation and application, ZL201310511154.2). Further activity tests show that the designed phenylquinolinone and flavonoid derivatives are dual-target compounds that antagonize H ₃ receptors and inhibit Aβ aggregation.

SUMMARY OF THE INVENTION

Glossary: The term "alkyl" as used herein, unless a different number of atoms is specified, refers to a straight or branched hydrocarbon chain containing 1-6 carbon atoms. Examples of "alkyl" as used herein 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 halogen or hydroxy.

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 t-butoxy. "Alkoxy" also includes substituted alkoxy. The alkoxy group can be optionally substituted one or more times with halogen.

"Pharmaceutically acceptable salt" in the present invention includes hydrochloride, sulfate, hydrobromide, (+) tartrate, (-) tartrate, fumarate, succinate, etc. commonly used in pharmacy Inorganic and organic acid salts, amino acid salts.

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

General structure (A)

where:

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, hydroxyl 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 with a total of 3-6 carbon atoms, including but not limited to the following fragments:

选自以下两种杂环：Further, the preferred quinolinone and flavonoid derivatives of the present invention, the

Selected from the following two heterocycles:

Further, in the preferred quinolinone and flavonoid derivatives and their analogs of the present invention, R ₁ is preferably hydrogen, methyl and ethyl; R ₂ is preferably hydrogen, hydroxyl and methoxy; R ₃ is preferably hydrogen , hydroxyl and methoxy.

It is to be understood that this invention includes all combinations and subgroups of the specific groups defined herein, including substitutions defined in the brief description above, exemplified in the various examples throughout the specification, and recited in the appended claims base.

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

Preferred compounds of table 1 quinolinone derivatives

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

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

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

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

Synthetic methods of I-3 to I-6 in the series of compound I (quinolinone derivatives) (this method is suitable when X=N):

The concrete reaction process is: the intermediate 4 in the embodiment 1 is dissolved in DMF, sodium hydride and halogenated hydrocarbons are added, and the reaction obtains the intermediate 5, which is finally reacted with secondary amine and triethylamine to obtain the target compound 1-3. ~I-6.

Synthetic method of I-7 and I-8 in compound I (quinolinone derivatives) series (this method is suitable when X=N):

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

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

Concrete reaction process is: intermediate 9, dimethyl sulfate and potassium carbonate in embodiment 3 are dissolved in acetone, and backflow reaction obtains intermediate 10, and then with secondary amine NHR'R" and triethylamine backflow reaction to obtain Target compound I-9, I-10.

Synthetic method of II-1 and II-2 in compound II (flavonoid derivatives) series (this method is suitable when X=O):

The specific reaction process is as follows: the raw material 6 is dissolved in acetonitrile, after adding 1,3-bromochloropropane and potassium carbonate, the intermediate 11 is obtained by refluxing reaction, and the latter is condensed with p-hydroxyacetophenone in potassium hydroxide to obtain The intermediate 12 is then cyclized through the hydrogen oxide/KOH system to obtain the intermediate 13, which is finally refluxed with secondary amine and triethylamine to obtain the target compounds II-1 and II-2.

Synthetic method of II-3 and II-4 in compound II (flavonoid derivatives) series (this method is suitable when X=O)

Concrete reaction process is as follows: intermediate 13, methyl iodide and potassium carbonate in embodiment 5 are dissolved in acetone, and backflow reaction obtains intermediate 14, and then with secondary amine, triethylamine backflow reaction to obtain target compound II-3, II-4.

Synthetic method of II-5 and II-6 in compound II (flavonoid derivatives) series (this method is suitable when X=O)

The specific reaction process is as follows: the intermediate 11 and 2,6-dihydroxyacetophenone are dissolved in ethanol, potassium hydroxide is added, and the reflux reaction is performed to obtain the intermediate 15. Intermediate 16 is finally refluxed with secondary amine and triethylamine to obtain target compounds II-5 and II-6.

Synthetic method of II-7 and II-8 in compound II (flavonoid derivatives) series (this method is suitable when X=O)

The concrete reaction process is as follows: the intermediate 16 of embodiment 7, potassium iodide and potassium carbonate are dissolved in acetone, and the backflow reaction obtains the intermediate 17, and then with secondary amine, triethylamine backflow reaction 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 their pharmaceutically acceptable salts and solvates in the preparation of medicines, the medicines are prepared from the analogs and pharmaceutically acceptable salts and solvates thereof. Acceptable salts and solvates are prepared with pharmaceutically acceptable carriers or excipients.

The "pharmaceutically acceptable carrier" refers to the conventional pharmaceutical carriers 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 bentonite, lubricants such as talc, etc., if necessary Flavoring agents, sweeteners, etc. may also be added.

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

The present invention provides the compounds and preferred compounds thereof, pharmaceutically acceptable salts of the compounds, solvates, prodrugs (esters or phosphates), stereoisomers, deuterated compounds and other compounds of the compounds Use of drug combination in the treatment of neurodegenerative related diseases. Wherein the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, narcolepsy, epilepsy, ALS, etc., particularly discloses the protection of the compounds of the present invention and their preferred compounds against ischemic brain injury It is not only conducive to the recovery of neurological function, but also effectively reduces the volume of cerebral infarction.

Description of drawings

Figure 1 shows some H3 receptor antagonists entering clinical research.

Figure 2 shows the design ideas of target molecules.

Figure 3 shows the effect of compounds on A[beta] self-aggregation.

In Fig. 3, [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, 24h. PBS pH 7.4.

Figure 4 shows the effect of compounds on Aβ depolymerization.

In Figure 4, [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, 24h. PBS pH 7.4, 50,000x.

Detailed ways

The specific examples below are included for purposes of illustration and should not be construed as limiting the scope of the invention. In addition, it should be understood that after reading the content taught by 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.

Example 1: Synthesis of I-1, I-2 in class I compounds (quinolinones).

Step one: 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 obtain a colorless oily liquid. Directly add 15 mL of 6N NaOH solution and 30 mL of CH ₃ OH, and reflux for 1 h. After the reaction solution became clear, cooled, acidified with 2N hydrochloric acid to pH=2, a large amount of white solid was precipitated, suction filtered, washed with water, and dried to obtain 14.5 g of white solid powder. Yield 94%; ¹ H NMR (500 MHz, 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=6.5Hz, 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 (5.0 g, 23 mmol) was refluxed in 10 mL of SOCl ₂ for 1 h, and 1 to 2 drops of DMF was added to the reaction system. After the reaction, excess SOCl ₂ was removed under reduced pressure to obtain intermediate 2 as a colorless liquid. 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. The filtrate was spin-dried, and the obtained residue was separated by column chromatography (petroleum ether:EtOAc=10:1) to obtain 5.0 g of a white solid. 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＝8.0,1.5Hz,1H),7.64(t,J＝8.0Hz,1H),7.16(t,J＝8.0Hz,1H),7.02(d,J＝9.0Hz,2H),4.21( t, J=6.0Hz, 2H), 3.78 (t, J=6.5Hz, 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 of THF, and the reaction was microwaved at 110° C. for 20 min in a closed vessel. After the reaction, cooled to room temperature, poured into 100 mL of ice water, added 2N HCl to adjust the pH to 5-6, a large amount of yellow solid was precipitated, washed with water, 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＝7.5Hz,1H),7.35(t,J＝7.5Hz,1H),7.17(d,J＝9.0Hz,2H),6.33(s ,1H),4.21(t,J＝6.0Hz,2H),3.84(t,J＝6.5Hz,2H),2.24-2.18(m,2H); ESI-MS: m/z＝314[M+H ] ⁺ .

Step four: 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, 41 mg (0.57 mmol) of pyrrolidine and 96 mg (0.96 mmol) of TEA were added dropwise, and the temperature was raised to reflux for overnight reaction. After the reaction was completed, it was cooled, the solvent was evaporated under reduced pressure, and separated by column chromatography (petroleum ether:EtOAc:TEA=1:5:0.1) to obtain 40 mg of yellow solid. 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＝8.5Hz, 2H), 7.34(t, J＝7.5Hz, 1H), 6.76(d, J＝8.5Hz, 2H), 6.38(s, 1H), 3.91(t, J=6.0Hz, 2H), 2.95-2.88 (m, 6H), 2.12-2.07 (m, 2H), 1.98-1.93 (m, 2H); ¹³ C NMR (125 MHz, 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] ⁺ .

2-(4-(3-(Piperidin-1-yl)propoxy)phenyl)quinolin-4(1H)-one (I-2).

The preparation method is the same as that of compound I-1, except that piperidine is used instead of pyrrolidine to obtain a yellow solid. Yield 75%; ¹ H NMR (500 MHz, 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＝8.5Hz, 2H), 7.34(t, J＝7.5Hz, 1H), 6.84(d, J＝8.5Hz, 2H), 6.39(s, 1H), 3.95(t, J=6.0Hz, 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 (125MHz, CDCl ₃ ): δ174.16, 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, 19.23, 20.5; 363[M+H] ⁺ .

Example 2: Synthesis of I-3 to I-6 in class I compounds (quinolinones).

Step one: 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, 42 mg of NaH (60%, 1.05 mmol) were added, and after stirring at room temperature for 30 min, methyl iodide (138 mg, 0.97 mmol) was added, and the temperature was raised to 35°C, The reaction was carried out for 30 minutes. The reaction solution was poured into 50 mL of H ₂ O, extracted with EtOAc, washed with water, washed with saturated NaCl, and dried over anhydrous Na ₂ SO ₄ . After recovering the solvent, it was separated by column chromatography (petroleum ether:EtOAc=10:1) to obtain 250 mg of white solid. Yield 87%; ¹ H NMR (500 MHz, 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＝7.5Hz, 1H), 7.14(s, 1H), 7.05(d, J＝9.0Hz, 2H), 4.21(t, J＝6.0Hz, 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] ⁺ .

2-(4-(3-Chloropropoxy)phenyl)-1-ethylquinolin-4(1H)-one (Intermediate 5b).

The preparation method is the same as that of compound intermediate 5a, and bromoethane is used instead of iodomethane to obtain a yellow solid. Yield 67%; ¹ HNMR (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＝7.5Hz, 1H), 7.12(s, 1H), 7.05(d, J＝9.0Hz, 2H), 4.37(q, J＝7.0Hz, 2H), 4.21(t, J＝6.0Hz, 2H), 3.79 (t, J=6.5Hz, 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 that of compound I-1, but intermediate 4 is replaced by intermediate 5a, and the secondary amine is pyrrolidine to obtain white solid I-3 with a yield of 74%; ¹ H NMR (500 MHz, CDCl ₃ ): δ8.17 (d, J＝8.0Hz, 1H), 8.09-8.06(m, 3H), 7.70(t, J＝8.0, 1H), 7.47(t, J＝7.5Hz, 1H), 7.14(s, 1H), 7.04(d,J＝9.0Hz,2H),4.13(t,J＝6.0Hz,2H),4.12(s,3H),2.71(t,J＝6.5Hz,2H),2.61-2.57(m,4H) ), 2.10-2.05 (m, 2H), 1.85-1.80 (m, 4H); ¹³ C NMR (100MHz, CDCl ₃ ): δ162.70, 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] ⁺ .

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

The preparation method is the same as that of compound I-1, except that intermediate 5a is used instead of intermediate 4, and piperidine is used for secondary amine instead of pyrrolidine to obtain white solid I-4 with a yield of 71%; ¹ H NMR (500 MHz, CDCl ₃ ): δ8.17(d,J＝8.0Hz,1H),8.09-8.06(m,3H),7.70(d,J＝8.0,1H),7.47(t,J＝7.5Hz,1H),7.14(s, 1H), 7.04(d, J=9.0Hz, 2H), 4.12(s, 3H), 4.11(t, J=6.0Hz, 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 (100MHz, CDCl ₃ ): δ162.70, 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] ⁺ .

2-(4-(3-(pyrrolidin-1-yl)propoxy)phenyl)-1-ethylquinolin-4(1H)-one (I-5).

The preparation method is the same as that of compound I-1, except that intermediate 5b is used instead of intermediate 4, and the secondary amine is pyrrolidine to obtain light 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=7.5Hz, 1H), 7.12 (s, 1H), 7.04 (d, J=9.0Hz, 2H), 4.37 (q, J=7.0Hz, 2H), 4.13 (t, J=6.0Hz) ,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=7.0Hz,3H )； ¹³ C NMR(100MHz,CDCl ₃ ):δ161.98,160.17,158.35,149.25,132.85,129.77,128.96,128.78,124.86,121.69,120.26,114.67,97.95,66.53,63.97,54.27,53.16,28.82,23.46, 14.54; ESI-MS: m/z=377 [M+H] ⁺ .

2-(4-(3-(Piperidin-1-yl)propoxy)phenyl)-1-ethylquinolin-4(1H)-one (I-6).

The preparation method is the same as that of compound I-1, except that intermediate 5b is used instead of intermediate 4, and the secondary amine is piperidine to obtain light 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=7.5Hz, 1H), 7.12 (s, 1H), 7.04 (d, J=9.0Hz, 2H), 4.37 (q, J=7.0Hz, 2H), 4.11 (t, J=6.0Hz) ,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=7.0Hz,3H _The ^_ , 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 and I-8 in class I compounds (quinolinones).

Step one: 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, the solvent was evaporated under reduced pressure, and separated by column chromatography (petroleum ether: EtOAc=10:1) to obtain 7.6 g of a colorless liquid. Yield 98%; ¹ H NMR (500 MHz, 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 (2.12 g, 10 mmol) was dissolved in 20 mL of diethyl ether, and Br ₂ (0.51 mL, 10 mmol) was slowly added dropwise at 0° C., the drop was completed, and the mixture was stirred at room temperature for 16 h. The reaction solution was poured into saturated NaHCO ₃ solution, extracted with ether, the organic layer was dried over anhydrous Na ₂ SO ₄ , and separated by column chromatography (petroleum ether: CH ₂ Cl ₂ =3:1) to obtain a pale yellow solid 2.45 g. Intermediate 84%; ¹ H NMR (500 MHz, 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 of DMF, stirred at room temperature for 30 min, and then intermediate 7 (1.46 g, 5.0 mmol) was added, and the temperature was raised to 50 ° C After 3 hours of reaction, the reaction solution was poured into 100 mL of water, extracted with EtOAc, the organic layer was washed with 1N NaOH solution and saturated NaCl solution, dried, evaporated under reduced pressure, and purified by column chromatography (petroleum ether: EtOAc = 3: 1) to obtain 1.6 g of white solid. 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＝8.5Hz,2H),6.72-6.68(m,2H),5.49(s,2H),4.22(t,J＝6.0Hz,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 the reaction, the reaction solution was poured into 250 mL of water, a large amount of solid was precipitated, suction filtered, washed with water until neutral, and dried to obtain 1.02 g of a light yellow solid. Yield 67%; ¹ H NMR (500 MHz, 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=7.5Hz,1H),6.97-6.95(m,2H),6.72-6.68(m,2H),4.12(t,J=6.0Hz,2H),3.76(t,J=6.0Hz,2H) , 2.26-2.23 (m, 2H); ESI-MS: m/z=330 [M+H] ⁺ .

Step five, 2-(4-(3-(pyrrolidin-1-yl)propoxy)phenyl)-3-hydroxyquinolin-4(1H)-one (I-7).

The preparation method is the same as that of compound I-1, and intermediate 9 is used instead of intermediate 4 to obtain a beige solid. Yield 59%; ¹ H NMR (500 MHz, 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 ＝9.0Hz, 2H), 4.12(t, J＝6.5Hz, 2H), 2.56(t, J＝7.0Hz, 2H), 2.46-2.42(m, 4H), 1.95-1.89(m, 2H), 1.70 -1.67 (m, 4H); ¹³ C NMR (125 MHz, 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, 54.1.3,6,6 52.68, 28.63, 23.58; ESI-MS: m/z=365 [M+H] ⁺ .

2-(4-(3-(pyrrolidin-1-yl)propoxy)phenyl)-3-hydroxyquinolin-4(1H)-one (I-8).

The preparation method is the same as compound I-1, using intermediate 9 instead of intermediate 4 and piperidine instead of pyrrolidine to obtain a yellow solid. Yield 67%; ¹ H NMR (500 MHz, 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 ＝9.0Hz, 2H), 4.12(t, J＝6.5Hz, 2H), 2.56(t, J＝7.0Hz, 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 ): δ 170.25, 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 and I-10 in class I compounds (quinolinones).

Step one, 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 the reaction was completed, cooling and suction filtration, the filtrate was recovered under reduced pressure and the solvent was purified by column chromatography (petroleum ether: EtOAc = 1:2) to obtain 558 mg of a yellow solid. Yield 78%; ¹ H NMR (500 MHz, 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＝7.5Hz, 1H), 7.31(d, J＝8.5Hz, 2H), 7.07(d, J＝8.5Hz, 2H), 4.22(t, J＝6.0Hz, 2H), 3.81(t, J=6.0Hz, 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 method is the same as compound I-1, and intermediate 10 is used instead of intermediate 4 to obtain an off-white solid. Yield: 65%; ¹ HNMR (500 MHz, 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＝7.5Hz, 1H), 7.28(d, J＝8.0Hz, 2H), 7.05(d, J＝8.5Hz, 2H), 4.12(t, J＝6.0Hz, 2H), 3.65(s, 3H), 3.53(s, 3H), 2.70(t, J=7.5Hz, 2H), 2.61-2.55(m, 4H), 2.13-2.05(m, 2H), 1.85-1.81 (m, 4H); ¹³ C NMR (125 MHz, 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, 66.0, 6,59 37.12, 28.74, 23.47; ESI-MS: m/z=393 [M+H] ⁺ .

2-(4-(3-(Piperidin-1-yl)propoxy)phenyl)-3-methoxy-1-methylquinolin-4(1H)-one (I-10).

The preparation method is the same as compound I-1, using intermediate 10 instead of intermediate 4 and piperidine instead of pyrrolidine to obtain an off-white solid. Yield: 67%; ¹ H NMR (500 MHz, 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＝7.5Hz,1H),7.29(d,J＝8.5Hz,2H),7.05(d,J＝8.5Hz,2H),4.10(t,J＝6.0Hz,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 _The ^_ , 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 and II-2 in class II compounds (flavonoids).

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

The preparation method is the same as that of intermediate 6, except that p-hydroxybenzaldehyde is used instead of p-hydroxyacetophenone to obtain a pale yellow solid. Yield: 98%; ¹ H NMR (500 MHz, 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) were dissolved in 15 mL of C ₂ H ₅ OH, 1.68 g of KOH (30 mmol) was added, and the reaction was refluxed for 2 h. After cooling, part of the solvent was removed under reduced pressure, 200 mL of ice water was added to the remaining residue, and the pH was adjusted to 4-5 with 2N HCl. Yield 89%; ¹ HNMR (500 MHz, 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＝7.5Hz,1H),7.03(d,J＝8.5Hz,1H),6.97-6.93(m,3H),4.20(t,J＝6.0Hz,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 0.6 mL of H ₂ O ₂ aqueous solution (30%) was added in batches, and after stirring at room temperature for 1 h, The reaction solution was poured into ice water, and a large amount of solid was precipitated, which was filtered off with suction, and the filter cake was washed with water and dried to obtain 930 mg of yellow solid. Yield 94%; ¹ H NMR (500 MHz, 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＝7.0Hz,1H),7.01(d,J＝9.0Hz,2H),4.15(t,J＝6.0Hz,2H),3.83(t,J＝6.5Hz,2H),2.23- 2.17 (m, 2H); ESI-MS: m/z=331 [M+H] ⁺ .

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

The preparation method is the same as that of compound I-1, and intermediate 13 is used instead of intermediate 4 to obtain a yellow solid. Yield: 68%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.25-8.21 (m, 3H), 7.71 (t, J=7.5Hz, 1H), 7.59 (d, J=8.5Hz, 1H) ,7.42(t,J＝7.5Hz,1H),7.06(d,J＝9.0Hz,2H),4.14(t,J＝6.0Hz,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] ⁺ .

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

The preparation method is the same as compound I-1, using intermediate 13 instead of intermediate 4 and piperidine instead of pyrrolidine to obtain a yellow solid. Yield: 61%; ¹ H NMR (500 MHz, CDCl ₃ ): δ 8.25-8.21 (m, 3H), 7.71 (t, J=7.5Hz, 1H), 7.59 (d, J=8.5Hz, 1H) ,7.42(t,J＝7.5Hz,1H),7.06(d,J＝9.0Hz,2H),4.12(t,J＝6.5Hz,2H),2.56(t,J＝7.0Hz,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=38 ⁺ ] .

Example 6: Synthesis of II-3 and II-4 in class II compounds (flavonoids).

Step one: 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 the reaction was completed, cooling and suction filtration, the filtrate was recovered under reduced pressure and the solvent was purified by column chromatography (petroleum ether: EtOAc = 1:2) to obtain 558 mg of a yellow solid. Yield 78%; ¹ H NMR (500 MHz, 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＝7.5Hz, 1H), 7.31(d, J＝8.5Hz, 2H), 7.07(d, J＝8.5Hz, 2H), 4.22(t, J＝6.0Hz, 2H), 3.81(t, J=6.0Hz, 2H), 3.65(s, 3H), 3.54(s, 3H), 2.36-2.24(m, 2H); ESI-MS: m/z=358[M+ H] ⁺ .

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

The preparation method is the same as compound I-1, and intermediate 14 is used instead of intermediate 4 to obtain a red solid. Yield: 54%; ¹ H NMR (500 MHz, 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=8.5Hz,2H),4.12(t,J=6.0Hz,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, ESI-MS: m/z ⁼ 380

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

The preparation method is the same as compound I-1, using intermediate 14 instead of intermediate 4 and piperidine instead of pyrrolidine to obtain a yellow solid. Yield: 57%; ¹ H NMR (500 MHz, 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=8.5Hz, 2H), 4.12(t, J=6.5Hz, 2H), 3.89(s, 3H), 2.56( t, J=7.0Hz, 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,CDCl ₃ ):δ175.00,161.03,155.71,155.16,140.82,133.27,130.24,125.80,124.57,124.22,123.09,117.90,114.51,66.65,59.93,55.89,54.67,26.68,25.90,24.38；ESI-MS: m/z=394 [M+H] ⁺ .

Example 7: Synthesis of II-5 and II-6 in class II compounds (flavonoids).

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

The preparation method is the same as that of intermediate 12, except that 2,6-dihydroxyacetophenone is used instead of o-hydroxyacetophenone to obtain 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 (1.00 g, 3.2 mmol) was dissolved in 15 mL of DMSO, I ₂ (128 mg, 0.5 mmol) and 0.5 mL of concentrated sulfuric acid were added, and the mixture was stirred at 85° C. for 24 h. The reaction solution was poured into 200 mL of H ₂ O, extracted three times with EtOAc, the organic layers were combined, washed with water, saturated NaCl, dried over anhydrous Na ₂ SO ₄ , and separated by column chromatography (petroleum ether: EtOAc = 4:1 ) to obtain 565 mg of white solid. Yield: 45%; 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＝8.0Hz,1H),6.82(d,J＝8.0Hz,1H),6.66(s,1H),4.23(t,J＝6.0Hz,2H),3.79(t , J=6.0 Hz, 2H), 2.32-2.27 (m, 2H); ESI-MS: m/z=331 [M+H] ⁺ .

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

The preparation method is the same as that of compound I-1, and intermediate 16 is used instead of intermediate 4 to obtain 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＝8.0Hz,1H),6.82(d,J＝8.0Hz,1H),6.66(s,1H),4.12(t,J＝6.0Hz,2H),2.78(t , J=6.0Hz, 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] ⁺ .

2-(4-(3-(Piperidin-1-yl)propoxy)phenyl)-5-hydroxy-4H-benzopyran-4-one (II-6).

The preparation method is the same as compound I-1, using intermediate 16 instead of intermediate 4 and piperidine instead of pyrrolidine to obtain a yellow solid. Yield: 64%; ¹ H NMR (500 MHz, 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＝8.0Hz,1H),6.81(d,J＝8.0Hz,1H),6.65(s,1H),4.13(t,J＝6.0Hz,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 and II-8 in class II compounds (flavonoids).

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

The preparation method is the same as that of intermediate 14, and intermediate 16 is used instead of intermediate 13 to obtain an off-white solid. Yield: 75%; ¹ HNMR (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), 6.99(d, J＝8.0Hz, 1H), 6.82(d, J＝8.0Hz, 1H), 6.68(s, 1H), 4.21(t, J＝6.0Hz, 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 two: 2-(4-(3-(pyrrolidin-1-yl)propoxy)phenyl)-5-methoxy-4H-benzopyran-4-one (II-7).

The preparation method is the same as compound I-1, and intermediate 17 is used instead of intermediate 4 to obtain a yellow solid. Yield: 61%; ¹ H NMR (500 MHz, 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＝8.0Hz,1H),6.82(d,J＝8.0Hz,1H),6.68(s,1H),4.13(t,J＝6.0Hz,2H),3.91(s , 3H), 2.76(t, J＝6.0Hz, 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] ⁺ .

2-(4-(3-(Piperidin-1-yl)propoxy)phenyl)-5-methoxy-4H-benzopyran-4-one (II-8).

The preparation method is the same as compound I-1, using intermediate 17 instead of intermediate 4 and piperidine instead of pyrrolidine to obtain a yellow solid. Yield: 67%; ¹ H NMR (500 MHz, 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＝8.0Hz,1H),6.83(d,J＝8.0Hz,1H),6.66(s,1H),4.12(t,J＝6.0Hz,2H),3.89(s ESI -MS: m/z=394[M+H] ⁺ .

Example 9: Histamine _H3 receptor antagonistic activity of phenylquinolinones and flavonoid derivatives.

In this part, with Clobenpropit as the positive control, LANCE Ultra TR-FRET method and ThT fluorescence assay were used to screen the histamine _H3 receptor antagonistic activity of the synthesized 18 target compounds. Other compounds of the present invention were the same as those listed below. The compounds have similar beneficial effects, but this should not be construed to mean that the compounds of the present invention have only the following beneficial effects. The results are shown in Table 3.

Table ₃ Histamine H3 receptor antagonistic activities of phenylquinolinone and flavonoid derivatives

From the receptor antagonistic activities in Table 3, it can be seen that the selected phenylquinolinone compounds exhibit strong histamine _H3 receptor inhibitory activity, most of the compounds have similar activities to Clobenpropit, and six molecules have more activity than Clobenpropit. good.

Example 10: Determination of anti-amyloid peptide aggregation activity.

A ThT fluorescence test.

(1) Solution and sample preparation: PBS buffer preparation (10mM, pH 7.4): Weigh 57.996g Na ₂ HPO ₄ ·12H ₂ O, 5.928g NaH ₂ PO ₄ ·2H ₂ O and dissolve in 1000 mL of water to prepare the concentration It is 0.2M PBS stock solution; take 50mL stock solution, add water to dilute to 1000mL, and filter with 0.22μm microporous membrane. Preparation of Gly-NaOH buffer solution (50mM, pH 8.5): Weigh 3.7535g of glycine and dissolve it in 800mL of water, slowly add 0.1M NaOH solution dropwise to adjust the pH to 8.5, and add water to make up to 1000mL. Preparation of Aβ monomer: 1 mg of Aβ _1-42 was dissolved in 1 mL of HFIP, ultrasonically shaken for 10 min, divided into packages, placed at room temperature for 12 h, dried in vacuum, and stored in a -20°C refrigerator. Aβ monomer was reconstituted to 2 mM in DMSO, and after brief vortexing, a 50 μM stock solution was prepared in 10 mM PBS buffer. Compound formulation: Compounds were dissolved in DMSO to prepare stock solutions at a concentration of 10 mM. Dilute to corresponding concentration with PBS. ThT solution preparation (5 μM): Weigh 1.59 g of ThT and dissolve it in 100 mL of Gly-NaOH buffer to prepare a stock solution with a concentration of 50 mM, and store in the dark. Take 10 μL of the stock solution and dilute it to 100 mL with Gly-NaOH buffer to obtain a 5 μM ThT solution.

(2) Add Aβ _1-42 and compounds (positive control or PBS buffer) to the EP tube to make the final concentrations of 25 μM and 20 μM, respectively, place at 37° C. with constant temperature shaking (150 rpm), and incubate for 24 h. Add 20 μL of the incubation solution to the 96-well plate, then add 180 μL of 5 μM ThT solution, incubate in the dark for 5 min, and use a multi-plate reader to detect the fluorescence intensity of the sample at 485 nm under excitation at 450 nm.

The fluorescence value of Aβ _1-42 after incubation alone was used as a control. In order to avoid the interference of the fluorescence of the compound itself on the result, 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

It can be seen from the above experimental results that most of the designed and synthesized target molecules show good Aβ _1-42 self-aggregation activity, while the positive control Clobenpropit has no such activity.

Compounds I-6 and II-1 were further selected for their activities of inhibiting self-aggregation of Aβ _1-42 and promoting depolymerization of aggregated Aβ _1-42 samples by transmission electron microscopy.

Tested with 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 solution. Dilute 5 mL of the mother solution to 400 mL, slowly add 1 mM NaOH solution dropwise to adjust the pH to 6.6, and dilute to 500 mL.

Self-aggregation samples: Add Aβ _1-42 and compounds (positive control Curcumin or PBS buffer) to EP tubes to make the final concentrations of 25 μM and 20 μM, respectively, and place them at 37°C under constant temperature shaking (150 rpm) and incubate for 24 h.

Depolymerization samples after self-aggregation: add Aβ _1-42 (final concentration of 50 μM) to the EP tube, incubate at 37°C for 24 hours, add compounds (positive control Curcumin or PBS buffer) to make it interact with Aβ _1- The final concentrations of ₄₂ were 20 μM and 25 μM, respectively, and were incubated together for 24 h.

(2) Take 10 μL of the incubation solution and place it on the carbon-supported membrane copper mesh. After standing for 2 minutes, blot dry with filter paper from the side. After standing for 30 seconds, add 5 μL of 1% uranyl acetate solution dropwise to negatively stain the sample. After standing for 2 minutes Blot dry from the side with filter paper, dry and observe under a transmission electron microscope, and take pictures. The accelerating voltage is 80kV and the magnification is 50,000 times. The results are shown in Figure 3.

Compounds I-6 and II-1 were subjected to Aβ _1-42 aggregation inhibition experiments. As shown in Figure 3, compared with 0 hours (Figure 3a), Aβ monomers were able to significantly self-aggregate after 24 hours (Figure 3b). ), while both compounds were able to significantly inhibit Aβ self-aggregation (Fig. 3c, 3d).

Further depolymerization experiments are shown in Figure 4. The samples added with compounds I-6 and II-1 have obvious depolymerization effects, which can degrade the aggregated filaments into short rods or granules (Figure 4b, 4c), and their activity is excellent. to curcumin (Fig. 4d).

Example 11: Effects on ischemic brain injury.

Rats were anesthetized with 10% chloral hydrate (3.5ml/kg, intraperitoneal injection), supine and fixed, and a skin incision was made along the left side of the neck. Artery, incision at the bifurcation of external carotid artery and internal carotid artery, insert suture (diameter 0.25mm, length 18mm), ligate the incision, suture the skin, loosen the distal internal carotid artery clip. After 2 hours, gently lift the remaining thread to a slight resistance to achieve reperfusion of the middle cerebral artery, and the modeling is completed. Rats in the sham-operated group were not subjected to middle artery insertion after anesthesia, and the rest of the operations were the same. During the experiment, the body temperature was kept at 37±0.5℃ with a constant temperature blanket, and penicillin sodium was added dropwise to the wound after the operation. Rats that survived for 24 hours were randomly divided into a sham operation group, a model group and each test group. The test group was given the compounds of the present invention and grouped by the numbers corresponding to the compounds.

Each group was given intraperitoneal injection, and rats in the model group and sham-operated group were given an equal volume of normal saline, and 16 hours after the first administration, each group was given another intraperitoneal injection.

Nerve injury symptom scoring method, 0 points: normal behavior, with both forelimbs extending symmetrically to the ground when tail is lifted; 1 point: wrist bending of the contralateral forelimb when tail is lifted; 2 points: elbow bending of the contralateral forelimb when tail is lifted; 3 Score: internal rotation of the shoulder when the tail is lifted; tilt to the opposite side of the injury when walking; 4: no voluntary movement.

At the time of the first administration, 4 hours after administration, and 20 hours after administration, the rats were scored for the symptoms of nerve damage. After 24 hours after the first administration, the whole brain was quickly taken out after being sacrificed by severed neck, and placed in the rats. On ice, 3 hippocampi were randomly taken out from each group for the detection of histamine content, the remaining part was frozen at -20°C, and the brain was cut into 5 slices with a razor blade in the coronal plane, and the brain slices were placed in 1% TTC at room temperature and protected from light. Stain for 30 min and fix with 4% paraformaldehyde for 30 min. The slices were photographed and analyzed, and the cerebral infarct volume ratio (infarct volume/whole brain volume*100%) was calculated.

Table 5 Measurement results of nerve injury symptoms, histamine content in hippocampus, and cerebral infarction volume ratio in each group

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

The rats were scored by the behavioral grading method. After 4 hours of the first administration, the symptoms of nerve damage were significantly different between the compounds of the present invention and the model group. 20 hours after the first administration, the compounds of the present invention and the model There is a significant difference between the two groups, indicating that the compound of the present invention has a significant improving effect on the neurological function of ischemic brain injury rats; the histamine content in the hippocampus, the compound of the present invention is different from the model group, indicating that the present invention The compound can effectively increase the content of histamine in the brain of ischemic brain injury rats; the infarct volume ratio, the compound of the present invention and the model group have significant differences, indicating that the compound of the present invention can effectively reduce ischemic brain injury. Cerebral infarct volume in mice.

Example 12: Effects on mice with acute epilepsy induced by pentylenetetrazole.

Each group of mice was given the test drug of the present invention and the positive drug phenytoin sodium respectively, and the intragastric administration was carried out according to the volume of 0.1ml/10g. The model group was given the same volume of normal saline, and the positive control group was given phenytoin sodium at a dose of 50mg/kg , each dose of the test drug group was 50 mg/kg, and the test drug group was given the compounds of the present invention, and grouped by the numbers corresponding to the compounds.

After 1 hour of intragastric administration, 70 mg/kg of pentylenetetrazole was intraperitoneally injected in each group, and the latency time of myoclonus (LTMJ) was recorded.

The epileptic seizure criteria refer to the Racine criteria, and the timing ends when the facial muscles or the corners of the mouth twitch, one side limb twitches, rigidity or generalized limb twitch epilepsy has a generalized grand mal.

Table 6 Mice seizure latency measurement results (n=6)

level number of animals Incubation period (min) model group 8 3.56±0.88 control group 8 10.11±6.69 Group I-3 8 13.64±5.76 Group I-6 8 14.49±7.17 Group I-9 8 15.11±8.19 Group II-4 8 17.86±9.53 Group II-7 8 14.39±8.69

The results show that, compared with the model group, the compounds of the present invention can significantly delay the acute epileptic seizures in mice, and the latency period is prolonged by 3 to 5 times; The compounds of the present invention have significant antiepileptic effects.

Claims (9)

1. A phenylquinolinone and flavonoid derivative is a compound with the following general structure (A) or pharmaceutically acceptable salt thereof:

in the formula:

x is selected from NR₁Or O;

R₁selected from H, C_1-3Alkyl or C_1-3An alkoxy group;

R₂selected from H, hydroxy or C_1-3An alkoxy group;

R₃selected from H, hydroxy, C_1-3Alkyl or C_1-3An alkoxy group;

NR 'R' is selected from

2. Phenyl quinolinone and flavonoid derivatives according to claim 1, characterized in that: in the general structure (A), when X is NR₁When the compound is a benzoquinolinone compound with the following general structure (A1) or a pharmaceutically acceptable salt thereof:

in the formula:

R₁selected from H, C_1-3Alkyl or C_1-3An alkoxy group;

R₂selected from H, hydroxy or C_1-3An alkoxy group;

R₃selected from H, hydroxy, C_1-3Alkyl or C_1-3An alkoxy group;

NR 'R' is selected from

3. Phenyl quinolinone and flavonoid derivatives according to claim 2, characterized in that: in the general structure (A1), R₁Is selected from H or C_1-3An alkyl group; r₂Selected from H, hydroxy or C_1-3An alkoxy group; r₃Selected from H, hydroxy or C_1-3An alkoxy group;

NR 'R' is selected from

4. Phenyl quinolinone and flavonoid derivatives according to claim 1, characterized in that: in the general structure (A), when X is O, the benzoflavonoid compound or the pharmaceutically acceptable salt thereof has the following general structure (A2):

in the formula:

R₂selected from H, hydroxy or C_1-3An alkoxy group;

R₃selected from H, hydroxy, C_1-3Alkyl or C_1-3An alkoxy group;

NR 'R' is selected from

5. Phenyl quinolinone and flavonoid derivatives according to claim 4, characterized in that: in the general structure (A2), R₂Selected from H, hydroxy or C_1-3An alkoxy group; r₃Selected from H, hydroxyOr C_1-3An alkoxy group; NR 'R' is selected from

6. Phenyl quinolinone and flavonoid derivative according to any one of claims 1 to 5, characterized in that: is a compound having the structural formula set forth in the following list:

7. a pharmaceutical composition comprising as an active ingredient at least one compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof, together with at least one pharmaceutically acceptable carrier or excipient.

8. Use of a compound according to any one of claims 1 to 6 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the treatment of neurodegenerative diseases and ischemic brain injury diseases.

9. Use according to claim 8, characterized in that: the neurodegenerative disease is selected from Alzheimer's disease, Parkinson's disease, lethargy, epilepsy or gradually freezing disease.

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