Naphthalene chalcone compound as well as preparation method and application thereof

Abstract

The invention discloses a naphthalene type chalcone compound and a preparation method and application thereof, and relates to the technical field of medicinal chemistry, and the structural formula of the naphthalene type chalcone compound is shown in the specification. Wherein part of the compounds have a good inhibition effect on HDAC6 protein, can obviously inhibit proliferation of tumor cells, and can be used for developing tumor treatment drugs related to abnormal expression of HDAC6 activity.

Description

Technical Field

[0001] This invention relates to the field of medicinal chemistry, specifically to a naphthyl chalcone compound, its preparation method, and its application. Background Technology

[0002] Histone deacetylases (HDACs) are a class of enzymes, both histone and non-histone proteins, that remove acetyl groups from lysine residues in cells. They can directly alter chromosome structure and regulate gene expression. Overexpression of HDACs is closely related to gene expression, cell cycle arrest, cell differentiation, and cell death. HDAC6 is one of the most representative HDACs, and dysregulation of HDAC6 is associated with the malignant progression of various cancers, including leukemia, breast cancer, and ovarian cancer.

[0003] Chalcones belong to the α,β-unsaturated ketone class of compounds and possess various activities including anticancer, anti-inflammatory, and antibacterial activity. To date, several chalcone compounds have been used as drugs in clinical research, demonstrating potential for new drug development. Research and development of these compounds has become a hot topic in medicinal chemistry. Patent CN116496179A discloses an HDAC6 inhibitor containing a chalcone structure, among which compound RH20-18 exhibits the strongest inhibitory effect on the HDAC6 protein, with an IC50 value of [missing information]. 50 The value is 27.4 nM. Patent ZA202302614B discloses an HDAC6 inhibitor containing a chalcone structure, among which compound BZ-06 has the strongest inhibitory effect on HDAC6 protein, IC50 value. 50 The value is 4.0 nM. Patent CN119823002A discloses a dual inhibitor of VEGFR2-HDAC6 containing a chalcone fragment, in which compound MD1 exhibits the strongest inhibitory effect on HDAC6 protein, with an IC50 value of 4.0 nM. 50 The value is 890 nM. Structure-activity relationship studies show that the structure of the surface recognition region has a significant impact on activity.

[0004]

[0005] Based on the above research, the present invention aims to design and synthesize a novel naphthalene-type chalcone compound, with the expectation of further improving the inhibitory activity of the compound against HDAC6 protein, thereby enabling the preparation of drugs for the prevention and / or treatment of diseases related to abnormal expression of HDAC6 activity. Summary of the Invention

[0006] The technical problem to be solved by the present invention is to provide a naphthyl chalcone compound and a method for preparing the same. These compounds exhibit good inhibitory activity against HDAC6 protein and can be used to prepare drugs for the prevention and / or treatment of diseases related to abnormal expression of HDAC6 activity.

[0007] The technical problem to be solved by this invention is achieved by the following technical solution:

[0008] The first object of this invention is to provide a naphthyl chalcone compound or a pharmaceutically acceptable salt, isomer, hydrate, solvate, crystal form, or prodrug thereof, the structural formula of which is shown below:

[0009]

[0010] In Formula I and Formula II, R1 and R2 are each independently selected from at least one of hydrogen, alkyl, alkoxy, hydroxyl, ester, nitro, and cyano.

[0011] Furthermore, in Formula I and Formula II, R1 and R2 are each independently selected from at least one of hydrogen and alkoxy groups.

[0012] Furthermore, in Formula I, R1 is hydrogen and R2 is hydrogen or methoxy; in Formula II, both R1 and R2 are hydrogen.

[0013] In some specific embodiments, the structural formula of the naphthalene-type chalcone compound is shown below:

[0014] A second objective of this invention is to provide a method for preparing the aforementioned naphthalene-type chalcone compounds, comprising the following steps:

[0015] (1) 4-Formylbenzaldehyde reacts with naphthyl ethyl ketone via a Claisen-Schmidt reaction to give intermediate A;

[0016] (2) Intermediate A reacts with O-(tetrahydro-2H-pyran)-2-hydroxyamine via a condensation reaction to obtain intermediate B;

[0017] (3) Intermediate B undergoes a deprotection reaction to yield compound I;

[0018] (4) p-acetylbenzoic acid reacts with naphthaldehyde via a Claisen-Schmidt reaction to obtain intermediate C;

[0019] (5) Intermediate C reacts with O-(tetrahydro-2H-pyran)-2-hydroxyamine via a condensation reaction to obtain intermediate D;

[0020] (6) Intermediate D undergoes a deprotection reaction to give compound II.

[0021] The synthesis route is as follows:

[0022]

[0023] Furthermore, the Claisen-Schmidt reaction is carried out under alkaline conditions; the alkaline conditions are provided by at least one of sodium hydroxide, potassium hydroxide, lithium hydroxide, and barium hydroxide.

[0024] Further, the molar ratio of 4-formylbenzaldehyde to naphthyl ethyl ketone is 1:(1-2); the molar ratio of p-acetylbenzoic acid to benzaldehyde is 1:(1-2).

[0025] Furthermore, the condensation reaction is carried out in the presence of a condensing agent and an activator; the condensing agent includes, but is not limited to, at least one of dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide (DIC), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDCI); the activator includes, but is not limited to, at least one of 4-dimethylaminopyridine (DMAP) and 1-hydroxybenzotriazole (HOBt).

[0026] Further, the molar ratio of intermediate A to O-(tetrahydro-2H-pyran)-2-hydroxylamine is 1:(1~2); the molar ratio of intermediate C to O-(tetrahydro-2H-pyran)-2-hydroxylamine is 1:(1~2).

[0027] Furthermore, the deprotection reaction is carried out under acidic conditions; the acidic conditions are provided by at least one of hydrochloric acid, sulfuric acid, and trifluoroacetic acid.

[0028] A third object of the present invention is to provide the use of the said naphthoid chalcone compound or its pharmaceutically acceptable salts, isomers, hydrates, solvates, crystal forms, and prodrugs in the preparation of HDAC6 inhibitors.

[0029] A fourth object of the present invention is to provide a pharmaceutical composition comprising the said naphthyl chalcone compound or a pharmaceutically acceptable salt, isomer, hydrate, solvate, crystal form, or prodrug thereof.

[0030] Furthermore, the pharmaceutical composition also includes a pharmaceutically acceptable carrier or excipient.

[0031] A fifth object of the present invention is to provide the use of the said naphthyl chalcone compound or a pharmaceutically acceptable salt, isomer, hydrate, solvate, crystal form, prodrug, or pharmaceutical composition thereof in the preparation of a medicament for the prevention and / or treatment of diseases associated with abnormal expression of HDAC6 activity.

[0032] Furthermore, the diseases associated with abnormal HDAC6 activity expression include tumors, inflammation, autoimmune diseases, neurodegenerative diseases, and nervous system diseases.

[0033] The beneficial effects of this invention are as follows: This invention utilizes computer-aided drug design to replace the benzene ring with a naphthalene ring in the surface recognition region of HDAC6 inhibitors containing chalcone fragments, resulting in a series of naphthyl chalcone compounds; tests have shown that some compounds exhibit good inhibitory effects on HDAC6 protein and can significantly inhibit the proliferation of tumor cells, and can be used to develop tumor therapeutic drugs related to abnormal expression of HDAC6 activity. Attached Figure Description

[0034] Figure 1 For intermediate A-1 1 H NMR spectrum;

[0035] Figure 2 For intermediate B-1 1 H NMR spectrum;

[0036] Figure 3 For compound 1 1 H NMR spectrum;

[0037] Figure 4 For compound 1 13 C NMR spectrum;

[0038] Figure 5 For compound 2 1 H NMR spectrum;

[0039] Figure 6 For compound 2 13 C NMR spectrum;

[0040] Figure 7 For compound 3 1 H NMR spectrum;

[0041] Figure 8 For compound 3 13 C NMR spectrum;

[0042] Figure 9 For intermediate C-4 1 H NMR spectrum;

[0043] Figure 10 For compound 4 1 H NMR spectrum;

[0044] Figure 11 For compound 4 13 C NMR spectrum;

[0045] Figure 12 For compound 5 1 H NMR spectrum;

[0046] Figure 13 For compound 5 13 C NMR spectrum. Detailed Implementation

[0047] To make the technical means, creative features, objectives and effects of this invention easier to understand, the invention will be further described below with reference to specific embodiments and illustrations.

[0048] Example 1

[0049] Synthesis of intermediate A-1:

[0050]

[0051] 4-Formylbenzoic acid (2.0 mmol) and 1-naphthyl ethyl ketone (2.05 mmol) were added to methanol, followed by potassium hydroxide (6.0 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, hydrochloric acid was added to adjust the pH to below 3. The mixture was filtered, and the filter cake was washed with a mixture of methanol and water, dried, and a pale yellow solid was obtained in 89.2% yield with a melting point of 210.2–212.0 °C. 1 HNMR (400MHz, DMSO-d6) δ13.13 (s, 1H), 8.39-8.34 (m, 1H), 8.17 (d, J = 8.3Hz, 1H), 8. 04(dd,J=7.2,1.5Hz,2H),8.01-7.90(m,4H),7.75-7.66(m,2H),7.66-7.57(m,3H).

[0052] Synthesis of intermediate B-1:

[0053]

[0054] Intermediate A-1 (1.0 mmol), O-(tetrahydro-2H-pyran)-2-hydroxylamine (1.1 mol), and triethylamine (3.5 mmol) were added to dichloromethane, followed by the addition of EDCI (1.2 mmol) and HOBt (0.1 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was washed with water, and the dichloromethane was removed by vacuum distillation. Column chromatography (eluent: dichloromethane) was then performed to give a yellow solid in 71.2% yield with a melting point of 265.5–267.2 °C. 1H NMR (400MHz, DMSO-d6) δ11.75(s,1H),8.40-8.30(m,1H),8.17(d,J=8.3Hz,1H),8.05(t,J=6.7Hz,2H),7.92(d,J=8.0Hz,2 H),7.83(d,J=8.0Hz,2H),7.75-7.54(m,5H),5.01(s,1H),4.06(s,1H),3.58-3.49(m,1H),1.73(s,3H),1.62-1.48(m,3H).

[0055] Synthesis of Compound 1:

[0056]

[0057] Intermediate B-1 (0.200 g) was added to isopropanol (4 mL), followed by 6 mol / L hydrochloric acid (1 mL). The mixture was stirred at room temperature for 30 min. After the reaction was complete, 4 mL of water was added, the mixture was stirred, filtered, and dried to obtain an off-white solid with a yield of 84.2% and a melting point of 162.1–163.8 °C. 1 H NMR (400MHz, DMSO-d6) δ11.34(s,1H),9.12(s,1H),8.39-8.33(m,1H),8.17(d,J=8.2Hz,1H), 8.04(td,J=7.2,1.7Hz,2H),7.90(d,J=8.3Hz,2H),7.81(d,J=8.4Hz,2H),7.72-7.58(m,5H). 13 C NMR(101MHz,DMSO-d6)δ193.97,163.45,143.66,137.00,135.90,134.26,132.08,129.86,1 28.78,128.58,128.20,127.92,127.65,127.40,126.53,125.21,124.93.HR-MS(m / z),[M+H] + Calculated value: 318.1125, measured value: 318.1128.

[0058] Example 2

[0059] The synthesis method of compound 2 is the same as that in Example 1, except that 1-naphthyl ethyl ketone is replaced with 2-naphthyl ethyl ketone, yielding a white solid with a yield of 83.1% and a melting point of 186.0-187.1℃. 1H NMR (400MHz, DMSO-d6) δ11.37(s,1H),9.14(s,1H),8.97(s,1H),8.22(d,J=15.6Hz,1H),8.17(d,J=7.9Hz,1H),8.15(dd,J=8. 6,1.7Hz,1H),8.07(d,J=8.7Hz,1H),8.03(d,J=8.2Hz,3H),7.87(d,J=8.3Hz,2H),7.84(d,J=15.6Hz,1H),7.70-7.66(m,2H). 13 C NMR(101MHz,DMSO-d6)δ188.84,163.50,142.69,137.33,135.11,134.72,134.17,132.32,130.6 5,129.63,128.85,128.79,128.52,127.73,127.38,127.01,124.09,123.53.HR-MS(m / z),[M+H] + Calculated value: 318.1125, measured value: 318.1149.

[0060] Example 3

[0061] The synthesis method of compound 3 is the same as that in Example 1, except that 1-naphthyl ethyl ketone is replaced with 4-methoxynaphthyl ethyl ketone, yielding a pale yellow solid with a yield of 82.9% and a melting point of 179.3-180.7℃. 1 H NMR (400MHz, DMSO-d6) δ11.35(s,1H),9.13(s,1H),8.70(d,J=8.3Hz,1H),8.27(d,J=8.1Hz,1H),8.23(d,J=7.6Hz,1H),7.91(d, J=7.9Hz,2H),7.83(d,J=7.8Hz,2H),7.78(s,1H),7.72-7.62(m,2H),7.58(t,J=7.5Hz,1H),7.08(d,J=7.7Hz,1H),4.08(s,3H). 13 C NMR(101MHz,DMSO-d6)δ191.69,163.58,158.30,142.22,137.37,134.00,131.86,131.64,128.63,1 28.35,127.67,127.59,127.43,126.02,125.62,125.11,121.93,103.23,56.17.HR-MS(m / z),[M+H] +Calculated value: 348.1230, measured value: 348.1239.

[0062] Example 4

[0063] Synthesis of intermediate C-4:

[0064]

[0065] 4-Acetylbenzoic acid (2.0 mmol) and 1-naphthaldehyde (2.05 mmol) were added to methanol, followed by potassium hydroxide (6.0 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, hydrochloric acid was added to adjust the pH to below 3. The mixture was filtered, and the filter cake was washed with a mixture of methanol and water, dried, and a pale yellow solid was obtained in 90.1% yield with a melting point of 223.3–225.1 °C. 1 HNMR(400MHz,DMSO-d6)δ13.69(s,1H),8.59(d,J=15.4Hz,1H),8.27(td,J=8.1,1.5Hz,4H),8.15 -8.10(m,2H),8.07(d,J=8.2Hz,1H),8.02(d,J=2.1Hz,1H),8.01-7.97(m,1H),7.68-7.57(m,3H).

[0066] Synthesis of intermediate D-4:

[0067]

[0068] Intermediate C-4 (1.0 mmol), O-(tetrahydro-2H-pyran)-2-hydroxylamine (1.1 mol), and triethylamine (3.5 mmol) were added to dichloromethane, followed by the addition of EDCI (1.2 mmol) and HOBt (0.1 mmol). The mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was washed with water, and the dichloromethane was removed by vacuum distillation. Column chromatography (eluent: dichloromethane) was then performed to give a pale yellow solid in 69.4% yield.

[0069] Synthesis of compound 4:

[0070]

[0071] Intermediate D-4 (0.200 g) was added to isopropanol (4 mL), followed by 6 mol / L hydrochloric acid (1 mL). The mixture was stirred at room temperature for 30 min. After the reaction was complete, 4 mL of water was added, the mixture was stirred, filtered, and dried to obtain a pale yellow solid with a yield of 83.5% and a melting point of 183.4–185.0 °C. 1H NMR (400MHz, DMSO-d6) δ11.47(s,1H,NH),9.23(s,1H,NH),8.59(d,J=15.3Hz,1H),8.2 8(td,J=6.5,3.4Hz,4H),8.10-7.99(m,3H),7.96(d,J=8.1Hz,2H),7.69-7.56(m,3H). 13 C NMR(101MHz,DMSO-d6)δ188.78,163.36,140.43,139.46,136.67,133.40,131.26,131.19,131.0 9,128.86,128.72,127.41,127.36,126.39,125.89,125.73,124.44,123.00.HR-MS(m / z),[M+H] + Calculated value: 318.1125, measured value: 318.1138.

[0072] Example 5

[0073] The synthesis method of compound 5 is the same as that in Example 4, except that 1-naphthoaldehyde is replaced with 2-naphthoaldehyde, yielding an off-white solid with a yield of 84.1% and a melting point of 205.4-207.0℃. 1 H NMR(400MHz,DMSO-d6)δ11.47(s,1H,NH),9.23(s,1H,NH),8.36(s,1H),8.27(d, J=8.1Hz,2H),8.17-8.05(m,2H),8.03-7.88(m,6H),7.58(qd,J=6.7,3.4Hz,2H). 13 C NMR(101MHz,DMSO-d6)δ188.77,163.34,144.61,139.51,136.61,134.04,132.97,132.28,130.9 8,128.67,128.61,128.54,127.77,127.59,127.36,126.87,124.48,122.27.HR-MS(m / z),[M+H] + Calculated value: 318.1125, measured value: 318.1136.

[0074] Example 6

[0075] The inhibitory activity of the above compounds against HDAC6 protein was tested using the HDAC6 Fluorogenic Assay Kit according to the method specified in the kit. BZ-06, MD1, and Trichostatin A were used as positive controls. In black 96-well plates, 5 μL of 200 μM Fluorogenic HDAC substrate, 5 μL of 0.1% BSA solution, 30 μL of HDAC detection buffer, 5 μL of 7 ng / μL HDAC6 recombinant enzyme, and 5 μL of different concentrations of the drug were added, respectively. In the blank group, no HDAC6 recombinant enzyme was added, and 10% DMSO aqueous solution was used instead of the drug. In the control group, 10% DMSO aqueous solution was used instead of the drug. The plates were incubated at 37°C for 30 min, and then 50 μL of HDAC Developer was added to each well. The plates were incubated at room temperature for 15 min, and the absorbance was measured using a microplate reader at an excitation wavelength of 350 nm and an emission wavelength of 450 nm. The IC50 was calculated using GraphPad Prism 8.0 software. 50 The values ​​are shown in Table 1.

[0076] Table 1. Inhibitory activities of compounds 1–6 on HDAC6 protein.

[0077] compound <![CDATA[IC 50 (nM)]]> compound <![CDATA[IC 50 (nM)]]> 1 7.9±0.4 RH20-18 27.9±1.2 2 6.3±0.6 BZ-06 4.1±0.2 3 95.6±5.8 MD1 895±31 4 16.7±1.3 Trichostatin A 14.7±0.7 5 21.4±0.9

[0078] As shown in Table 1, except for compound 3, the other four compounds (1-5) exhibit strong inhibitory effects on HDAC6 protein, with IC50 values ​​of 1-5. 50 The values ​​were all below 50 nM and stronger than the control RH20-18; the inhibitory effects of compounds 1 and 2 were stronger than the positive control Trichostatin A and similar to the positive control BZ-06.

[0079] Example 7

[0080] The inhibitory effects of compounds 1–5 on chronic myeloid leukemia cells K562 and cervical cancer cells Siha were detected using the CCK8 assay.

[0081] Siha cells and K562 cells were cultured in PRIM1640 medium containing 1% penicillin and 10% serum at 37°C in a 5% CO2 cell culture incubator. Siha cells and K562 cells in the logarithmic growth phase were collected separately, and approximately 2 × 10⁶ cells were added to each well of a 96-well plate. 3 Cells were incubated for 12 hours, then different concentrations of compounds 1–5 were added, and incubation continued for 48 hours. Then, 10 μL of CCK8 solution (10 mg / mL) was added, and incubation continued for 2 hours. The absorbance of each well was measured at 450 nm using a microplate reader, and the IC50 was calculated using GraphPadPrism 8.0 software. 50Values. The results are shown in Table 2:

[0082] Table 2 shows the inhibitory activity of compounds 1–5 on Siha and K562 cells.

[0083]

[0084] Table 2 shows that compounds 1–5 exhibited strong inhibitory effects on cervical cancer cells (SiHa) and chronic myeloid leukemia cells (K562). Compounds 1 and 2 showed stronger inhibitory activities against both K562 cells and SiHa than the control SAHA, with compound 2 exhibiting the strongest inhibitory activity, reaching a half-maximal inhibitory concentration (IC50) against both K562 and SiHa cells. 50 The values ​​were 0.60 μM and 0.83 μM, respectively, which were also stronger than the positive controls RH20-18, BZ-06 and MD1.

[0085] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the present invention as claimed. The scope of protection of this invention is defined by the appended claims and their equivalents.

Claims

1. A naphthalene-type chalcone compound or a pharmaceutically acceptable salt, isomer, hydrate, solvate, crystal form, prodrug thereof, having a structural formula as shown in the following: wherein R1 and R2 are each independently selected from at least one of hydrogen, alkyl, alkoxy, hydroxyl, ester, nitro and cyano. comprising the following steps:

2. The method for preparing the naphthalene-type chalcone compound according to claim 1, characterized in that, (1) obtaining an intermediate A by Claisen-Schmidt reaction of 4-formylbenzaldehyde and acetophenone; (2) obtaining an intermediate B by condensation reaction of the intermediate A and O-(tetrahydro-2H-pyran)-2-hydroxylamine; (3) obtaining the compound of formula I by deprotection reaction of the intermediate B; (4) obtaining an intermediate C by Claisen-Schmidt reaction of p-acetylbenzoic acid and naphthaldehyde; (5) obtaining an intermediate D by condensation reaction of the intermediate C and O-(tetrahydro-2H-pyran)-2-hydroxylamine; (6) obtaining the compound of formula II by deprotection reaction of the intermediate D; The synthetic route is as shown in the following: The Claisen-Schmidt reaction is carried out under alkaline conditions; 3. The method for preparing naphthalene-type chalcone compounds according to claim 2, characterized in that: Preferably, the alkaline conditions are provided by at least one of sodium hydroxide, potassium hydroxide, lithium hydroxide and barium hydroxide; Preferably, the molar ratio of the 4-formylbenzaldehyde to the acetophenone is 1: (1-2) ; Preferably, the molar ratio of the p-acetylbenzoic acid to the benzaldehyde is 1: (1-2). The condensation reaction is carried out in the presence of a condensing agent and an activating agent; 4. The method for preparing naphthalene-type chalcone compounds according to claim 2, characterized in that: Preferably, the condensing agent is at least one of dicyclohexyl carbodiimide, diisopropyl carbodiimide and 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide; Preferably, the activating agent is at least one of 4-dimethylaminopyridine and 1-hydroxybenzotriazole. Preferably, the molar ratio of the intermediate A to the O-(tetrahydro-2H-pyran)-2-hydroxylamine is 1: (1-2) ; Preferably, the molar ratio of the intermediate C to the O-(tetrahydro-2H-pyran)-2-hydroxylamine is 1: (1-2). The deprotection reaction is carried out under acidic conditions; 5. The method for preparing naphthalene-type chalcone compounds according to claim 2, characterized in that: Preferably, the acidic conditions are provided by at least one of hydrochloric acid, sulfuric acid and trifluoroacetic acid. 6.The naphthalene-type chalcone compound or the pharmaceutically acceptable salt, isomer, hydrate, solvate, crystal form, prodrug thereof of claim 1 for use in the preparation of an HDAC6 inhibitor. 7.A pharmaceutical composition comprising the naphthalene-type chalcone compound or the pharmaceutically acceptable salt, isomer, hydrate, solvate, crystal form, prodrug thereof of claim 1. The pharmaceutical composition further comprises a pharmaceutically acceptable carrier or excipient.

8. The pharmaceutical composition of claim 7, wherein: 9.The naphthalene-type chalcone compound or the pharmaceutically acceptable salt, isomer, hydrate, solvate, crystal form, prodrug thereof of claim 1 or the pharmaceutical composition of any one of claims 7-8 for use in the preparation of a medicament for preventing and / or treating a disease associated with abnormal expression of HDAC6 activity. The disease associated with abnormal expression of HDAC6 activity includes tumor, inflammation, autoimmune disease, neurodegenerative disease and nervous system disease.

10. Use according to claim 9, characterized in that: ​

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