Chalcone mannich base compounds, pharmaceutical compositions, and methods of making and using the same

Abstract

The present application belongs to the field of medicinal chemistry and pharmacotherapy, and particularly relates to a chalcone mannich base compound, a pharmaceutical composition and a preparation method and application thereof. The present application takes paeonol as a raw material, and obtains corresponding 3-substituted and 5-amino-methyl-substituted paeonol mannich base through mannich reaction of paeonol, polyoxymethylene and different organic amines. After the 3-substituted and 5-substituted paeonol mannich base is separated through column chromatography, the chalcone mannich base compound is obtained through Claisen-Schmidt reaction of the 3-substituted and 5-substituted paeonol mannich base and benzaldehyde containing different substituents. Experiments prove that the chalcone mannich base compound improves the physical and chemical properties of the chalcone mannich base and improves the drug efficacy, can solve the problem of poor water solubility, and has good anti-tumor cell proliferation activity.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of pharmaceutical chemistry and pharmacotherapy, and particularly relates to a chalcone Mannich base compound, a pharmaceutical composition and a preparation method and application thereof. BACKGROUND

[0002] Malignant tumors have high morbidity and mortality, seriously threatening human life and health. With the continuous progress of science and technology, more and more means of treating tumors have been developed, and certain therapeutic effects have been achieved. At present, the main treatment method for malignant tumors is drug treatment, but its clinical effect is not very satisfactory, and there are many shortcomings such as poor selectivity, large toxic and side effects, and easy drug resistance, so it is urgent to develop effective, safe and low-toxicity drugs to meet the needs of tumor patients (Cancer Epidemiol., 2021, 149(4): 778-789; Nat Rev Clin Oncol, 2013, 10, 571-587.).

[0003] Natural chalcone compounds are a class of organic compounds containing a 1,3-diphenylpropenone skeleton, which are widely present in edible plants such as vegetables, fruits and tea, and medicinal plants such as safflower and licorice. The α,β-alkenone structure in the chalcone skeleton can covalently bind to the affinity group in the biological macromolecular compound as a Michael acceptor, showing various biological activities such as anti-tumor, anti-inflammatory, antioxidant, antibacterial, etc. (Med. Res. Rev., 2020, 40: 2049-2084; Curr. Top. Med. Chem., 2017, 17: 3146-3169; Aging, 2019, 11: 7805-7816.), which has attracted widespread attention in drug research and development. Through the principles and methods of drug design, chemical modification of the structure of chalcone can obtain new anti-tumor drugs with novel structure, higher activity and good drugability, which has high research and development value (Med. Chem., 2017, 60: 1734-1735; J. Med. Chem., 2003, 46: 2813-2815; Bioorg. Med. Chem. Lett., 2010, 20: 7205-7211.).

[0004] The applicant has previously used paeonol (2-hydroxy-4-methoxyphenylacetone) as a raw material to perform Claisen-Schmidt condensation with different substituted benzaldehydes using the acetyl group at position 1 to obtain corresponding chalcone derivatives (Journal of Shangqiu Teachers College, 2023, 39(06): 28-30.). In vitro anti-tumor activity screening results show that the obtained target has strong anti-tumor cell proliferation activity, but its water solubility is poor. SUMMARY

[0005] To solve the above problems, in a first aspect, the present application provides a class of chalcone Mannich base compounds, the base compound is a compound shown in formula I or II or a pharmaceutically acceptable salt thereof:

[0006]

[0007] wherein R1 is Cl, F, CH3, (CH3)2N;

[0008] R2 is NR 1 R 2 ; R 1 and R 2 may be the same or different and independently represent a hydrogen atom, a C1-C6 alkyl group, an aromatic hydrocarbon group, (CH2)n-NR 3 R 4 , wherein n = 1-6, or R 1 and R 2 together with the nitrogen atom to which they are attached form a five- to seven-membered aliphatic heterocyclic ring, the ring group can be optionally mono- to penta-substituted with the same or different substituents, which include: a C1-C6 alkyl group, an aromatic hydrocarbon group, a hydroxyl group or a hydroxyl-(C1-C6) alkyl group;

[0009] R 3 and R 4 may be the same or different and independently represent a hydrogen atom or a C1-C6 alkyl group.

[0010] Further, in formula I or II, R2 is an amino group, a 2-aminoethylamine group, a 2-ethylaminoethylamine group, an aniline group, a benzylamine group, a phenethylamine group, a dimethylamine group, a diethylamine group, a dipropylamine group, a di-n-butylamine group, a tetrahydropyrrole group, a piperidine group, a morpholine group, a piperazine group, a 4-methylpiperazine group, an N-methylpiperazine group, an N-ethylpiperazine group, an N-benzylpiperazine group or a 4-hydroxyethylpiperazine group.

[0011] Further, in formula I or II, R2 is a 2-ethylaminoethylamine group, a benzylamine group, a phenethylamine group, a dimethylamine group, a diethylamine group, a tetrahydropyrrole group, a piperidine group, a morpholine group, a piperazine group, an N-methylpiperazine group, an N-ethylpiperazine group, an N-benzylpiperazine group or a 4-hydroxyethylpiperazine group.

[0012] Further, the base compound is any one of the following compounds:

[0013] 4-chloro-2'-hydroxy-4'-methoxy-3'-[(2-ethylaminoethyl)amino-1-yl]methyl chalcone;

[0014] 4-chloro-2'-hydroxy-4'-methoxy-3'-benzylaminomethyl chalcone;

[0015] 4-chloro-2'-hydroxy-4'-methoxy-3'-dimethylaminomethylchalcone;

[0016] 4-chloro-2'-hydroxy-4'-methoxy-3'-diethylaminomethylchalcone;

[0017] 4-chloro-2'-hydroxy-4'-methoxy-3'-(piperidin-1-yl)methylchalcone;

[0018] 4-chloro-2'-hydroxy-4'-methoxy-3'-morpholinomethylchalcone;

[0019] 4-chloro-2'-hydroxy-4'-methoxy-3'-(piperazin-1-yl)methylchalcone;

[0020] 4-chloro-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methylchalcone;

[0021] 4-chloro-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methylchalcone;

[0022] 4-chloro-2'-hydroxy-4'-methoxy-3'-(benzylpiperazin-1-yl)methylchalcone;

[0023] 4-chloro-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methylchalcone;

[0024] 4-methyl-2'-hydroxy-4'-methoxy-3'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0025] 4-methyl-2'-hydroxy-4'-methoxy-3'-benzylaminomethylchalcone;

[0026] 4-methyl-2'-hydroxy-4'-methoxy-3'-dimethylaminomethylchalcone;

[0027] 4-methyl-2'-hydroxy-4'-methoxy-3'-diethylaminomethylchalcone;

[0028] 4-methyl-2'-hydroxy-4'-methoxy-3'-(piperidin-1-yl)methylchalcone;

[0029] 4-methyl-2'-hydroxy-4'-methoxy-3'-morpholinomethylchalcone;

[0030] 4-methyl-2'-hydroxy-4'-methoxy-3'-(piperazin-1-yl)methylchalcone;

[0031] 4-methyl-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methylchalcone;

[0032] 4-methyl-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-l-yl)methylchalcone;

[0033] 4-methyl-2'-hydroxy-4'-methoxy-3'-(benzylpiperazin-l-yl)methylchalcone;

[0034] 4-methyl-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-l-yl)methylchalcone;

[0035] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-[(2-ethylaminoethyl)amino-l-yl]methylchalcone;

[0036] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-benzylaminomethylchalcone;

[0037] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-dimethylaminomethylchalcone;

[0038] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-diethylaminomethylchalcone;

[0039] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(piperidin-l-yl)methylchalcone;

[0040] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-morpholinomethylchalcone;

[0041] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(piperazinyl-l-yl)methylchalcone;

[0042] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-l-yl)methylchalcone;

[0043] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-l-yl)methylchalcone;

[0044] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(benzylpiperazin-l-yl)methylchalcone;

[0045] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-l-yl)methylchalcone;

[0046] 4-fluoro-2'-hydroxy-4'-methoxy-3'-[(2-ethylaminoethyl)amino-l-yl]methylchalcone;

[0047] 4-fluoro-2'-hydroxy-4'-methoxy-3'-benzylaminomethylchalcone;

[0048] 4-fluoro-2'-hydroxy-4'-methoxy-3'-dimethylaminomethylchalcone;

[0049] 4-fluoro-2'-hydroxy-4'-methoxy-3'-diethylaminomethylchalcone;

[0050] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(piperidin-1-yl)methylchalcone;

[0051] 4-fluoro-2'-hydroxy-4'-methoxy-3'-morpholinomethylchalcone;

[0052] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(piperazin-1-yl)methylchalcone;

[0053] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methylchalcone;

[0054] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methylchalcone;

[0055] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(benzylpiperazin-1-yl)methylchalcone;

[0056] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methylchalcone;

[0057] 4-chloro-2'-hydroxy-4'-methoxy-5'-[(2-ethylaminoethyl)amino]methylchalcone;

[0058] 4-chloro-2'-hydroxy-4'-methoxy-5'-benzylaminomethylchalcone;

[0059] 4-chloro-2'-hydroxy-4'-methoxy-5'-dimethylaminomethylchalcone;

[0060] 4-chloro-2'-hydroxy-4'-methoxy-5'-diethylaminomethylchalcone;

[0061] 4-chloro-2'-hydroxy-4'-methoxy-5'-(piperidin-1-yl)methylchalcone;

[0062] 4-chloro-2'-hydroxy-4'-methoxy-5'-morpholinomethylchalcone;

[0063] 4-chloro-2'-hydroxy-4'-methoxy-5'-(piperazin-1-yl)methylchalcone;

[0064] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0065] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0066] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0067] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0068] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0069] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0070] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0071] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0072] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0073] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0074] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0075] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0076] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0077] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0078] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0079] 4-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0080] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-benzylaminomethylchalcone;

[0081] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-dimethylaminomethylchalcone;

[0082] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-diethylaminomethylchalcone;

[0083] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(piperidin-1-yl)methylchalcone;

[0084] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-morpholinomethylchalcone;

[0085] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(piperazin-1-yl)methylchalcone;

[0086] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-1-yl)methylchalcone;

[0087] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-1-yl)methylchalcone;

[0088] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(benzylpiperazin-1-yl)methylchalcone;

[0089] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-1-yl)methylchalcone;

[0090] 4-fluoro-2'-hydroxy-4'-methoxy-5'-[(2-ethylaminoethyl)amino-1-yl]methylchalcone;

[0091] 4-fluoro-2'-hydroxy-4'-methoxy-5'-benzylaminomethylchalcone;

[0092] 4-fluoro-2'-hydroxy-4'-methoxy-5'-dimethylaminomethylchalcone;

[0093] 4-fluoro-2'-hydroxy-4'-methoxy-5'-diethylaminomethylchalcone;

[0094] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(piperidin-1-yl)methylchalcone;

[0095] 4-fluoro-2'-hydroxy-4'-methoxy-5'-morpholinomethylchalcone;

[0096] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(piperazin-1-yl)methylchalcone;

[0097] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-1-yl)methylchalcone;

[0098] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-1-yl)methylchalcone;

[0099] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(benzylpiperazin-1-yl)methylchalcone;

[0100] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-1-yl)methylchalcone.

[0101] Still further, the basic compound is any one of the following compounds:

[0102] 4-chloro-2'-hydroxy-4'-methoxy-3'-morpholinomethylchalcone;

[0103] 4-chloro-2'-hydroxy-4'-methoxy-3'-(piperidin-1-yl)methylchalcone;

[0104] 4-chloro-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methylchalcone;

[0105] 4-chloro-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methylchalcone;

[0106] 4-chloro-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methylchalcone;

[0107] 4-methyl-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methylchalcone;

[0108] 4-methyl-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methylchalcone;

[0109] 4-methyl-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methylchalcone;

[0110] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methylchalcone;

[0111] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methylchalcone;

[0112] 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methyl chalcone;

[0113] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methyl chalcone;

[0114] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methyl chalcone;

[0115] 4-fluoro-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methyl chalcone;

[0116] 4-chloro-2'-hydroxy-4'-methoxy-5'-morpholinomethyl chalcone;

[0117] 4-chloro-2'-hydroxy-4'-methoxy-5'-(piperidin-1-yl)methyl chalcone;

[0118] 4-chloro-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-1-yl)methyl chalcone;

[0119] 4-chloro-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-1-yl)methyl chalcone;

[0120] 4-chloro-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-1-yl)methyl chalcone;

[0121] 4-methyl-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-1-yl)methyl chalcone;

[0122] 4-methyl-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-1-yl)methyl chalcone;

[0123] 4-methyl-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-1-yl)methyl chalcone;

[0124] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-1-yl)methyl chalcone;

[0125] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-1-yl)methyl chalcone;

[0126] 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-1-yl)methyl chalcone;

[0127] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-1-yl)methyl chalcone;

[0128] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-1-yl)methyl chalcone;

[0129] 4-fluoro-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-1-yl)methyl chalcone;

[0130] In a second aspect, the present application provides a preparation method of the chalcone Mannich base compound, and a synthetic route thereof is as follows:

[0131]

[0132] In condition i, the reaction system further comprises anhydrous ethanol and 50% NaOH, and the reaction temperature is 50°C; wherein, the molar ratio of raw material 1 to raw material 2 is 1:1, the mass-volume ratio of raw material 1 to anhydrous ethanol is 0.83 g:20 mL, and the mass-volume ratio of raw material 1 to NaOH is 0.83 g:1.5 mL.

[0133] In condition ii, the reaction system further comprises 95% ethanol and paraformaldehyde, and the reaction temperature is 85°C; wherein, the molar ratio of intermediate 3 to paraformaldehyde is 1:6, and the mass-volume ratio of intermediate 3 to 95% ethanol is 288 mg:10 mL.

[0134] Further, the preparation method comprises the following steps:

[0135] Si: anhydrous ethanol is added to raw material 1 and raw material 2, and stirred at 50°C until the solid is completely dissolved to obtain a reaction solution; 50% NaOH solution is added dropwise to the reaction solution, and continues to be stirred until the reaction is complete; after cooling to room temperature, 6 mol / L dilute hydrochloric acid is added to adjust the pH to 6, a yellow precipitate is generated, and after filtration, the yellow precipitate is recrystallized by 95% ethanol to obtain intermediate 3;

[0136] Sii: 95% ethanol is added to intermediate 3 and paraformaldehyde, and stirred at 85°C until the solid is completely dissolved, then raw material 3 is added, and continues to be refluxed until the reaction is complete; after the reaction is complete, the organic phase is extracted by dichloromethane, concentrated under reduced pressure, and purified by column chromatography to obtain compounds I and II; wherein, the eluent used in column chromatography is methanol / dichloromethane with a volume ratio of 1:40;

[0137] The raw material 1 is paeonol, and the raw material 2 is a para-substituted benzaldehyde, including 4-chlorobenzaldehyde, 4-fluorobenzaldehyde, 4-methylbenzaldehyde or 4-dimethylaminobenzaldehyde.

[0138] The raw material 3 is any one of morpholine, piperidine, N-methylpiperazine, N-ethylpiperazine or N-(2-hydroxyethyl)piperazine, and the molar ratio of raw material 3 to intermediate 3 is 6:1.

[0139] In a third aspect, the present application provides a pharmaceutical composition comprising a therapeutically effective amount of the chalcone mannich base compound or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.

[0140] Further, the pharmaceutical composition further comprises at least one therapeutic agent; and the pharmaceutical composition is in any clinically or pharmaceutically acceptable dosage form.

[0141] The pharmaceutical composition can be used in the preparation of a medicament for treating osteosarcoma, lung cancer and liver cancer.

[0142] The pharmaceutical composition can be used in the preparation of a medicament for treating cervical cancer, colon cancer, osteosarcoma, lung cancer and liver cancer, wherein the pharmaceutical composition comprises a therapeutically effective amount of any one of the following compounds:

[0143] The compounds are: 4-chloro-2'-hydroxy-4'-methoxy-3'-morpholinomethylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-5'-morpholinomethylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-3'-(piperidin-l-yl)methylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-5'-(piperidin-l-yl)methylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-l-yl)methylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-l-yl)methylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-l-yl)methylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-l-yl)methylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-l-yl)methylchalcone, 4-chloro-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-l-yl)methylchalcone, 4-methyl-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-l-yl)methylchalcone, 4-methyl-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-l-yl)methylchalcone, 4-methyl-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-l-yl)methylchalcone, 4-methyl-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-l-yl)methylchalcone, 4-methyl-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-l-yl)methylchalcone, 4-methyl-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-l-yl)methylchalcone, 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-l-yl)methylchalcone, 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-l-yl)methylchalcone, 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-l-yl)methylchalcone, 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(ethylpiperazin-l-yl)methylchalcone, 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-l-yl)methylchalcone, or 4-dimethylamino-2'-hydroxy-4'-methoxy-5'-(hydroxyethylpiperazin-l-yl)methylchalcone.

[0144] Advantages of the present invention:

[0145] The present application takes paeonol as raw material, and through Mannich reaction with paraformaldehyde and different organic amines, corresponding 3-substituted and 5-amino methyl substituted paeonol Mannich base is obtained, after 3-substituted and 5-substituted paeonol Mannich base is separated through column chromatography, and then respectively with benzaldehyde containing different substituents, chalcone Mannich base compound is obtained through Claisen-Schmidt reaction, and experiments prove that the chalcone Mannich base compound provided in the present application improves the physical and chemical properties of chalcone Mannich base and improves the drug efficacy, and can solve the problem of poor water solubility, and the chalcone Mannich base compound provided in the present application has good anti-tumor cell proliferation activity.

[0146] Other features and advantages of the present application will be set forth in the following description, and in part will become apparent to those skilled in the art from the description, or can be learned by practice of the present application. The objects and other advantages of the present application will be realized and attained by the structure particularly pointed out in the description, claims and drawings. BRIEF DESCRIPTION OF DRAWINGS

[0147] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description are some embodiments of the present application, and for those skilled in the art, other drawings can also be obtained without creative labor.

[0148] Figure 1 The synthesis route diagram of the chalcone Mannich base compound in the embodiment of the present application is shown. DETAILED DESCRIPTION

[0149] In order to make the objects, technical solutions and advantages of the embodiments of the present application more clear, the following will combine the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are some embodiments of the present application, but not all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor are within the protection scope of the present application.

[0150] The present application introduces different amine methyl groups on the chalcone A ring through Mannich reaction, and designs and synthesizes a series of chalcone Mannich base derivatives.

[0151] The preparation method of the chalcone Mannich base derivative provided in the present application is:

[0152] The intermediate 3- hydroxy-4-methoxy-4'-substituted chalcone is obtained by Claisen-Schmidt reaction using paeonol as raw material 1 and p-substituted benzaldehyde 2 as starting material; the corresponding chalcone Mannich base (I1-14, II1-14) is obtained by Mannich reaction of the intermediate 3, paraformaldehyde and different organic amines.

[0153] The synthetic route is shown in Figure 1 The conditions i in the figure are that the reaction system further includes anhydrous ethanol, 50% NaOH, and the reaction temperature is 50°C; wherein the molar ratio of raw material 1 to raw material 2 is 1:1, the mass-volume ratio of raw material 1 to anhydrous ethanol is 0.83g:20mL, and the mass-volume ratio of raw material 1 to NaOH is 0.83g:1.5mL;

[0154] In the conditions ii, the reaction system further includes 95% ethanol and paraformaldehyde, and the reaction temperature is 85°C; wherein the molar ratio of intermediate 3 to paraformaldehyde is 1:6, and the mass-volume ratio of intermediate 3 to 95% ethanol is 288mg:10mL.

[0155] R1 represents Cl, F, CH3, (CH3)2N;

[0156] R2 represents NR 1 R 2 ; R 1 and R 2 may be the same or different and independently represent a hydrogen atom, a C1-C6 alkyl group, an aromatic hydrocarbon group, a (CH2)n-NR 3 R 4 , wherein n=1-6, or R 1 and R 2 together with the nitrogen atom to which they are attached form a five- to seven-membered aliphatic heterocyclic ring, which ring group can be optionally mono- to penta-substituted with the same or different substituents including a C1-C6 alkyl group, an aromatic hydrocarbon group, a hydroxyl group or a hydroxyl-(C1-C6) alkyl group; R 3 and R 4 may be the same or different and independently represent a hydrogen atom or a C1-C6 alkyl group.

[0157] The detailed preparation process includes the following steps:

[0158] S1: Anhydrous ethanol is added to raw material 1 paeonol and raw material 2, and stirred at 50°C until the solid is completely dissolved to obtain a reaction solution; 50% NaOH solution is slowly added dropwise to the reaction solution, and stirring is continued until the reaction is complete; after cooling to room temperature, 6mol / L dilute hydrochloric acid is added to adjust the pH to 6, a yellow precipitate is generated, and after filtration, the yellow solid powder intermediate 3 is obtained by recrystallization from 95% ethanol;

[0159] In step S1, thin layer chromatography (TLC) is used to monitor whether the reaction is complete; raw material 1 is paeonol, and raw material 2 is para-substituted benzaldehyde, including 4-chlorobenzaldehyde, 4-fluorobenzaldehyde, 4-methylbenzaldehyde or 4-dimethylaminobenzaldehyde; the molar ratio of raw material 1 and raw material 2 is 1:1, the volume ratio of anhydrous ethanol and raw material 2 is 20-25:1, and the volume ratio of NaOH and raw material 2 is 2.5:1.

[0160] S2: 95% ethanol is added to intermediate 3 and paraformaldehyde, and stirred at 85°C until the solid is completely dissolved. Then raw material 3 is added, and the reaction is continued under reflux. TLC is used to monitor until the reaction is complete. The organic phase is extracted with dichloromethane, concentrated under reduced pressure, and purified by column chromatography (eluent: methanol / dichloromethane, volume ratio 1:40) to obtain target compounds I1 and II1.

[0161] In step S2, the molar ratio of intermediate 3 and paraformaldehyde is 1:6, the mass-volume ratio of intermediate 3 and 95% ethanol is 288mg:10mL, and the molar ratio of raw material 3 and intermediate 3 is 6:1, wherein the raw material 3 is different organic amine compounds, including but not limited to morpholine, piperidine, N-methylpiperazine, N-ethylpiperazine or N-(2-hydroxyethyl)piperazine.

[0162] The above-mentioned intermediates or target compounds can be purified according to conventional separation techniques, and converted into addition salts with pharmaceutically acceptable acids, including hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, acetic acid, lactic acid, malonic acid, succinic acid, glutaric acid, fumaric acid, tartaric acid, maleic acid, citric acid, ascorbic acid, methanesulfonic acid, camphoric acid, oxalic acid, etc.

[0163] The structure of the chalcone Mannich base derivative, the preparation method thereof and the specific application effect thereof are described in detail below in combination with specific examples.

[0164] Example 1: Preparation of I1 and II1

[0165] Preparation of 4-chloro-2'-hydroxy-4'-methoxy chalcone (3a)

[0166] S1: 0.83g (5mmol) of paeonol and 0.7g (5mmol) of p-chlorobenzaldehyde are taken in a 50mL round-bottom flask, 20mL of anhydrous ethanol is added, and stirred at 50°C until the solid is completely dissolved. Then 1.5mL of 50% NaOH solution is slowly added dropwise to the reaction solution, and the stirring reaction is continued. TLC is used to monitor until the reaction is complete. After cooling to room temperature, 6mol / L dilute hydrochloric acid is added to adjust pH to 6, and yellow precipitate is generated. After filtration, 95% ethanol is used for recrystallization to obtain 1.208g of yellow solid powder (intermediate 3a), with a yield of 83.90% and a melting point of 111.5-112.4°C.

[0167] S2: Synthesis of target compounds I1and II1

[0168] Take 288 mg (1 mmol) of intermediate 3a, 0.18 g (6 mmol) of polyformaldehyde into a 50 mL round-bottom flask, add 10 mL of 95% ethanol, stir at 85°C until the solid is completely dissolved, add 0.52 mL (6 mmol) of morpholine, continue refluxing the reaction, monitor the reaction by TLC until it is complete, extract the organic phase with dichloromethane, concentrate under reduced pressure, and purify by column chromatography (eluent methanol / dichloromethane in a volume ratio of 1:40) to obtain compounds I1and II1.

[0169] The prepared solid is subjected to nuclear magnetic resonance hydrogen spectrum and mass spectrometry detection, and the detection results of compound I1are as follows:

[0170] 4-chloro-2'-hydroxy-4'-methoxy-3'-morpholinylmethyl chalcone (I1): yellow solid, yield 37.50%, melting point 155.7-156.6°C;

[0171] 1 H NMR (600 MHz, CDCl3) δ: 8.26 (d, J = 9.0, 1H), 8.02 (d, J = 15.6 Hz, 1H), 7.92 (d, J = 8.2 Hz, 2H), 7.76 (d, J = 15.6 Hz, 1H), 7.51 (d, J = 8.2 Hz, 2H), 6.68 (d, J = 9.0 Hz, 1H), 3.87 (s, 3H), 3.52 (s, 2H), 3.39 (d, J = 5.0 Hz, 4H), 2.42-2.36 (m, 4H); 13 C NMR (151 MHz, CDCl3) δ: 190.47, 163.02, 162.60, 141.33, 135.33, 132.46, 130.20, 128.62, 128.21, 121.29, 114.78, 111.11, 101.14, 65.92, 54.88, 52.33; HRMS, m / z: C 21 H 22 ClNO4, [M+H] + : Theoretical value 388.1316, measured value 388.1302.

[0172] 4-chloro-2'-hydroxy-4'-methoxy-5'-morpholinylmethyl chalcone (II1): yellow solid powder, yield 39.71%, melting point 160.5-161.3°C;

[0173] 1H NMR (600 MHz, DMSO-d6) δ: 13.34 (s, 1H), 8.09 (s, 1H), 7.96 (d, J = 15.4 Hz, 1H), 7.92 (d, J = 8.1 Hz, 2H), 7.78 (d, J = 15.4 Hz, 1H), 7.52 (d, J = 8.0 Hz, 2H), 6.55 (s, 1H), 3.84 (s, 3H), 3.68 (s, 2H), 3.53 (q, J = 4.7 Hz, 4H), 2.36 (t, J = 4.6 Hz, 4H). 13 C NMR (151 MHz, CDC13) δ: 191.61, 166.12, 164.73, 142.89, 136.59, 133.33, 131.84, 129.74, 129.32, 120.92, 113.47, 99.53, 66.97, 55.87, 55.82, 53.47; HRMS, m / z: C 21 H 22 ClNO4, [M+H] + : Calcd 388.1316, Found 388.1303.

[0174] Preparation of I6 and II6 in Example 2

[0175] Preparation of 4-methyl-2'-hydroxy-4'-methoxychalcone (3b)

[0176] S1: Take 0.83 g (5 mmol) of paeonol, 0.6 mL (5 mmol) of p-tolualdehyde, and add 20 mL of anhydrous ethanol to a 50 mL round-bottom flask, and stir at 50°C until the solid is completely dissolved. Slowly add 1.5 mL of 50% NaOH solution to the reaction solution, and continue to stir and react. Monitor the reaction completion by thin layer chromatography (TLC). After cooling to room temperature, add 6 mol / L dilute hydrochloric acid to adjust the pH to 5-6, and a yellow precipitate is generated. After filtration, recrystallize the yellow solid powder 1.147 g (intermediate 3b) obtained by 95% ethanol to obtain a yield of 85.62%, and the melting point is 118.7-120.6°C.

[0177] S2: Synthesis of target compounds I6 and II6

[0178] Take 288 mg (1 mmol) of intermediate 3b, 180 mg (6 mmol) of paraformaldehyde, and add 10 mL of 95% ethanol to a 50 mL round-bottom flask, and stir at 85°C under reflux until the solid is completely dissolved. Add 0.6 mL (6 mmol) of methylpiperazine, and continue to reflux and react. Monitor the reaction completion by TLC, extract the organic phase with dichloromethane, and concentrate under reduced pressure. Purify by column chromatography (eluent methanol / dichloromethane volume ratio 1:40) to obtain compounds I6 and II6.

[0179] The prepared solid was subjected to nuclear magnetic resonance hydrogen spectrum and mass spectrometry detection, and the results were as follows:

[0180] 4-Methyl-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methyl chalcone (I6): yellow solid powder, yield: 36.2%, melting point 137.1-138.5°C;

[0181] 1 H NMR (600MHz, CDCl3) δ7.90(d,J=8.8Hz,2H),7.82(d,J=15.4Hz,1H),7.60-7.57(m,3H),7.21(d,J=15.4 Hz,1H),6.73(d,J=8.2Hz,1H),3.87(s,3H),3.80(s,2H),2.88–2.55(m,8H),2.43(s,3H),2.38(s,3H); 13 C NMR (151MHz, CDCl3) δ193.56,163.79,163.15,142.41,139.82,133.66,130.09,129.7 8,129.57,119.33,116.51,111.51,100.65,56.48,55.71,54.01,52.66,45.61,21.40. HRMS, m / z: C 23 H 29 N2O3,[M+H] + : Theoretical value 381.2178, measured value 381.2685.

[0182] 4-Methyl-2'-hydroxy-4'-methoxy-5'-(methylpiperazin-1-yl)methyl chalcone (II6): yellow solid powder, yield: 38.8%, melting point 138.7-140.1°C;

[0183] 1 H NMR(600MHz,Chloroform-d)δ13.47(s,1H),7.88(s,1H),7.86(d,J=15.5Hz,1H),7.62–7.54(m,3H),7.25(s,1 H),7.23(d,J=15.5Hz,1H),6.44(s,1H),3.85(s,3H),3.52(s,2H),2.57-2.50(m,8H)2.40(s,3H),2.30(s,3H). 13C NMR (151 MHz, CDC13) δ 192.04, 165.90, 164.48, 144.28, 141.14, 132.19, 131.70, 129.71, 128.58, 119.58, 117.64, 113.54, 99.39, 55.72, 55.30, 55.15, 52.85, 45.98, 21.49. HRMS, m / z: C 23 H 29 N2O3, [M+H] + : Calcd 381.2178, Found 381.2685.

[0184] I2and II2, I3and II3, I4and II4, I5and II5, I7and II7, I8and II8, I9and II9, I 10 and II 10 , I 11 and II 11 , I 12 and II 12 , I 13 and II 13 , I 14 and II 14 were prepared according to the procedures described in Examples 1 and 2, respectively. The starting materials and final products used in each example are shown in Table 1:

[0185] Table 1

[0186]

[0187]

[0188]

[0189] Test Example 1

[0190] The in vitro anti-cell proliferation activities of compounds I 1-14 , II 1-14 on human cervical cancer cells (Hela), human colon cancer cells (HT29), human liver cancer cells (HepG2), human lung cancer cells (A549) and human osteosarcoma cells (U2OS) were tested by MTT method using the clinically commonly used anti-tumor drug fluorouracil (5-FU) as a positive control.

[0191] Experimental method:

[0192] The cells used were routinely cultured in a 37°C, 5% CO2saturated humidity incubator. The cancer cells in logarithmic growth phase with good growth state were added with 0.25% trypsin for digestion to make the adherent cells fall off. The cells were counted to be (2-3) x 10 4The cells were inoculated in 96-well plates at about 5000 cells per well, and incubated in an incubator for 24 hours. The culture solution was replaced with the tested drug (0.1-25.0 μmol / L, 5 concentrations) in an amount of 100 μL per well, and the incubation was continued in a 37°C, 5% CO2 saturated humidity incubator for 48 hours. MTT reagent was added to the 96-well plates in an amount of 20 μL per well, and incubated in the incubator for 4 hours. The supernatant was removed, DMSO was added in an amount of 120 μL per well, and the plates were shaken on a plate shaker for 10 minutes. The absorbance at a wavelength of 495 nm was measured by an enzyme-linked immunoassay instrument. All tests were repeated 3 times under the same conditions, and the cell inhibition rate was calculated.

[0193] Cell inhibition rate = [(OD value of the negative control group - OD value of the tested group) / (OD value of the negative control group - OD value of the blank group)] * 100%;

[0194] The IC50 was calculated by the method of probability unit weighted regression (Bliss method) using SPSS (Staffstical Package for the Social Science) 17.0 50 Some experimental results are shown in Table 2.

[0195] Table 2

[0196]

[0197]

[0198] As shown in the above table, the I1-I 11 and II1-II 11 have inhibitory effects on the proliferation of human cervical cancer cells (Hela), human colon cancer cells (HT29), human liver cancer cells (HepG2), human lung cancer cells (A549) and human osteosarcoma cells (U2OS), have good anti-tumor cell proliferation activity, and overall, the anti-tumor activity of most of the compounds is better than that of the clinical anti-tumor drug 5-FU. 12 -I 14 and II 12 -II 14 have inhibitory effects on the proliferation of human liver cancer cells (HepG2), human lung cancer cells (A549) and human osteosarcoma cells (U2OS), have good anti-tumor cell proliferation activity.

[0199] Although the present application has been described in detail with reference to the foregoing embodiments, it should be understood by those skilled in the art that modifications can be made to the technical solutions described in the foregoing embodiments, or some of the technical features can be replaced by equivalent features; and such modifications or replacements do not cause the essence of the corresponding technical solution to deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A chalcone Mannich base compound, characterized in that: The base compound is a compound represented by Formula I or a pharmaceutically acceptable salt thereof: wherein R1 is Cl, F, CH3 or (CH3)2N; R2 is selected from any one of morpholinyl, N-methylpiperazinyl, N-(2-hydroxyethyl)piperazinyl and N-ethylpiperazinyl.

2. The chalcone Mannich base compound according to claim 1, wherein: In formula I, R2 is selected from any one of N-methylpiperazinyl, N-ethylpiperazinyl and N-(2-hydroxyethyl)piperazinyl.

3. The chalcone Mannich base compound according to claim 1, wherein: The alkaline compound is any of the following compounds: 4-chloro-2'-hydroxy-4'-methoxy-3'-morpholinomethyl chalcone; 4-chloro-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methyl chalcone; 4-chloro-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methyl chalcone; 4-chloro-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methyl chalcone; 4-methyl-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methyl chalcone; 4-methyl-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methyl chalcone; 4-methyl-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methyl chalcone; 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methyl chalcone; 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methyl chalcone; 4-dimethylamino-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methyl chalcone; 4-Fluoro-2'-hydroxy-4'-methoxy-3'-(methylpiperazin-1-yl)methyl chalcone; 4-Fluoro-2'-hydroxy-4'-methoxy-3'-(ethylpiperazin-1-yl)methyl chalcone; 4-Fluoro-2'-hydroxy-4'-methoxy-3'-(hydroxyethylpiperazin-1-yl)methyl chalcone.

4. A method for preparing the chalcone Mannich base compound according to any one of claims 1 to 3, characterized in that: The preparation method The following steps are involved: Si: Anhydrous ethanol was added to raw materials 1 and 2, and the mixture was stirred at 50°C until the solid was completely dissolved to obtain a reaction solution; 50% NaOH solution was added dropwise to the reaction solution, and stirring was continued until the reaction was complete; after cooling to room temperature, 6 mol / L dilute hydrochloric acid was added to adjust the pH to 6, and a yellow precipitate was formed. After filtration, the mixture was recrystallized from 95% ethanol to obtain intermediate 3; Sii: 95% ethanol was added to the intermediate 3 and paraformaldehyde, and the mixture was refluxed and stirred at 85°C until the solid was completely dissolved. Then, the raw material 3 was added and the reflux reaction was continued until the reaction was complete. After the reaction was complete, the organic phase was extracted with dichloromethane, concentrated under reduced pressure, and purified by column chromatography to obtain compound I; the eluent used in the column chromatography was methanol / dichloromethane in a volume ratio of 1:40; Wherein, the raw material 1 is paeonol, and the raw material 2 is a para-substituted benzaldehyde selected from any one of 4-chlorobenzaldehyde, 4-fluorobenzaldehyde, 4-methylbenzaldehyde or 4-dimethylaminobenzaldehyde; The raw material 3 is selected from any one of morpholine, N-methylpiperazine, N-ethylpiperazine or N-(2-hydroxyethyl)piperazine, and the molar ratio of the raw material 3 to the intermediate 3 is 6:

1.

5. A pharmaceutical composition, characterized in that The pharmaceutical composition contains a therapeutically effective amount of the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers or excipients.

6. Use of the base compound according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 5 in the preparation of a drug for treating tumors, wherein the tumors include osteosarcoma, lung cancer and liver cancer.

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