阅读说明：本技术 一种联苯衍生物类化合物、制备方法及应用 (Biphenyl derivative compound, preparation method and application ) 是由 张厚巍 宫有文 于 2020-12-29 设计创作，主要内容包括：本发明属于医药领域,涉及一种联苯衍生物类化合物、制备方法及其应用。所述的联苯衍生物类化合物结构式如式I所示：药理学研究表明,本发明制备的化合物能够调节相关转录因子的功能,其中尤其是对于叉头转录蛋白O功能的调节,该化合物对于恶性肿瘤有明显的药物和治疗用途,尤其是对于肝细胞癌、乳腺癌等恶性肿瘤的治疗,抑制效果极佳。(The invention belongs to the field of medicines, and relates to a biphenyl derivative compound, a preparation method and application thereof. The structural formula of the biphenyl derivative compound is shown as the formula I: pharmacological research shows that the compound prepared by the invention can regulate the functions of related transcription factors, especially the function of forkhead transcription protein O, has obvious medicament and treatment application to malignant tumors, especially has excellent inhibition effect on the treatment of malignant tumors such as hepatocellular carcinoma, breast cancer and the like.)

1. A biphenyl derivative compound, the chemical structural general formula of which is shown in formula I,

in the formula: r₁Is selected from-OCH₃or-OEt, R₂Is selected from-OCH₃or-OEt.

2. The biphenyl derivatives of claim 1, formula I, is synthesized by the following steps:

3. the biphenyl derivative compounds according to claim 1, for use in the preparation of a medicament for treating diseases associated with deregulation of transcription factors or cofactors of transcription factors.

4. The use of claim 3, wherein the compound acts as a modulator of selective binding to a transcription factor or a co-factor of a transcription factor.

5. The use according to claim 4, wherein the transcription factor is forkhead transcription protein O and its isoforms.

6. The use according to claim 3, wherein the disease associated with dysregulation of a transcription factor or a co-factor of a transcription factor is a malignant tumor.

7. The use of claim 6, wherein the malignant tumor is breast cancer, hepatocellular carcinoma, glioblastoma, gastric cancer, colorectal cancer, breast cancer, or the like.

Technical Field

The invention belongs to the field of medicines, and relates to a biphenyl derivative compound, a preparation method and application thereof in preparation of transcription factors or medicines for diseases related to disregulation of auxiliary factors of the transcription factors.

Background

Cancer is one of the leading causes of human death, accounting for about one-sixth of the total annual death worldwide. The global cancer report shows that 1810 thousands of new cancer cases and 960 thousands of death cases are increased in 2018 all over the world, and the new cancer cases become important diseases threatening the health of human beings. Tumorigenesis is a complex process with multiple damage to the normal cellular genome. These lesions include activation of oncogenes, inactivation or deletion of oncogenes.

The FOX family is called fork-head box (fork-head box) proteins and is characterized by a DNA binding domain with a length of 110 amino acids, which is highly conserved and highly homologous to the corresponding region of the Drosophila fork gene. The sequences according to the forkhead box can be further divided into 19 subfamilies of FOXA-FOXS.

The forkhead transcription protein o (foxo) family is involved in the regulation of cell cycle progression, energy metabolism and tumorigenesis through specific activation of the transcription process. FOXO dysfunction can lead to uncontrolled cell proliferation and accumulation of DNA damage, leading to carcinogenesis. Expression and activity of FOXOs transcription factors are regulated by post-transcriptional modifications (including phosphorylation, methylation, acetylation, ubiquitination, miRNAs, etc.). It is considered as a tumor suppressor, which can activate or suppress the expression of downstream target genes and participate in various cell biological processes such as cell proliferation, apoptosis, differentiation, stress resistance and the like.

The invention innovatively synthesizes a biphenyl derivative compound which can be used as a transcription factor regulator, in particular to a regulator of forkhead transcription protein O, and can directly regulate the transcription factor so as to obtain a brand new compound which can be used for preparing medicines for diseases related to the imbalance of the transcription factor or the auxiliary factor of the transcription factor, and the prior art does not have reports of related structures.

Disclosure of Invention

In view of the above problems, the present invention aims to provide a preparation method and an application of biphenyl derivatives, which can regulate the functions of related transcription factors, especially the function of forkhead transcription protein O, and have obvious pharmaceutical and therapeutic applications for malignant tumors, especially for the treatment of malignant tumors such as hepatocellular carcinoma, breast cancer, etc., with excellent inhibitory effect.

In order to achieve the purpose, the invention adopts the following technical scheme.

A biphenyl derivative compound has a structural general formula I as follows:

in the formula: r₁Is selected from-OCH₃or-OEt, R₂Is selected from-OCH₃or-OEt.

Another objective of the present invention is to provide a synthetic route of biphenyl derivatives compound formula I:

of partial compounds¹H-NMR (400MHz) and¹³C-NMR (125MHz) was as follows:

compound 1:¹H-NMR(400MHz,CDCl₃)δ:2.60(t,2H),3.37(t,2H),3.75(s,3H),3.77(s,2H),3.83(s,3H),3.84(s,1H),4.02(s,1H),5.72(s,1H),6.60(d,1H),6.69(s,2H),6.74(s,1H),6.79(d,1H),7.72(t,2H),7.85(d,2H),8.11(s,2H),8.26(d,2H).¹³C-NMR(125MHz,CDCl₃)δ:35.88,41.98,43.58,56.83,110.20,113.55,113.87,122.42,128.57,129.72,129.82,130.59,132.91,132.94,133.10,136.72,137.77,147.86,148.22,150.28,168.07,169.19.

compound 2:¹H-NMR(400MHz,CDCl₃)δ:1.46(t,3H),2.67(t,2H),3.31(m,2H),3.77(d,2H),3.84(s,3H),3.99(t,1H),4.08(m,2H),6.22(s,1H),6.50(s,2H),6.60(m,1H),6.65(m,2H),6.77(d,1H),7.64(t,2H),7.86(m,2H),8.25(m,2H),8.50(t,2H),9.74(s,2H).¹³C-NMR(125MHz,CDCl₃)δ:14.80,34.40,41.11,43.18,56.32,64.46,113.03,114.52,116.85,122.26,127.94,130.21,130.37,131.30,132.15,135.77,146.58,147.16,149.22,155.35,168.00,169.56.

compound 3:¹H-NMR(400MHz,CDCl₃)δ:1.46(mt 6H),2.67(t,2H),3.31(m,2H),3.76(d,2H),3.99(d,1H),4.08(m,2H),4.16(m,2H),6.22(s,1H),6.50(s,2H),6.60(m,1H),6.65(t,1H),6.78(m,2H),7.62(t,2H),7.87(m,2H),8.25(m,2H),8.50(t,2H),9.74(s,2H).¹³C-NMR(125MHz,CDCl₃)δ:14.80,34.40,41.11,43.18,63.97,64.46,115.04,116.85,122.06,127.94,130.21,130.57,132.00,132.15,135.77,146.58,147.76,150.80,155.35,168.00,169.56.

the biphenyl derivative compound has a good weight-losing effect, can regulate the functions of related transcription factors, especially the function of forkhead transcription protein O, has obvious medicament and treatment application on malignant tumors, especially the treatment on malignant tumors such as hepatocellular carcinoma, breast cancer and the like, and has an excellent inhibiting effect. The biphenyl derivative compound has positive significance in treating malignant tumors and can be further researched.

The medicine for treating malignant tumor or the pharmaceutically acceptable salt or solvate thereof is applied to the preparation of medicines for treating diseases related to the imbalance of transcription factors or auxiliary factors of the transcription factors, and particularly provides the application of a biphenyl derivative compound in the preparation of medicines for treating malignant tumor such as breast cancer, hepatocellular carcinoma, glioblastoma, gastric cancer, colorectal cancer, breast cancer and the like.

Compared with the prior art, the invention has the following beneficial effects:

the compound prepared by the invention has good treatment effect in treating diseases related to disorder of the transcription factor, can regulate the function of the related transcription factor, especially for regulating the function of forkhead transcription protein O, and has obvious medicament and treatment application for malignant tumors, especially for treating malignant tumors such as hepatocellular carcinoma, breast cancer and the like. Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

Drawings

FIG. 1: the biphenyl derivative compounds obtained by the invention have influence on the proliferation of human cervical cancer cells.

FIG. 2: the biphenyl derivative compound obtained by the invention has influence on the expression of FOXOa protein.

Detailed Description

The following synthetic examples, biological test results, are used to further illustrate the invention, but are not meant to limit the invention.

Synthesis examples

Example 1 preparation of Compound 1

(1) Synthesis of 2' -methoxy-5 ' -nitro- [1,1':3',1' -terphenyl ] -3,3 "-dicarboxylic acid:

1, 3-dibromo-2-methoxy-5-nitrobenzene (12.18g, 0.039mol), 4-carboxyphenylboronic acid (13.61g, 0.082mol), 2M sodium carbonate solution (40.0mL) and tetrakis (triphenylphosphino) palladium (2.79g) were added to a dioxane (90.0mL) solution, stirred and heated to reflux under nitrogen for 24 hours. The reaction mixture was cooled and filtered, and the solid was slurried in 6M aqueous hydrochloric acid (100mL) for half an hour. Filtration and thorough washing of the filter cake with water followed by ether gave a dark yellow solid of 2' -methoxy-5 ' -nitro- [1,1':3',1' -terphenyl ] -3,3 "-dicarboxylic acid, 7.44g, 48.5% yield.

¹H-NMR(400MHz,CDCl₃)δ:4.16(s,3H),7.72(t,2H),7.85(d,2H),8.11(s,2H),8.26(d,2H),8.70(s,2H).¹³C-NMR(125MHz,CDCl₃)δ:60.70,124.51,128.45,130.29,130.83,132.64,137.09,137.28,138.11,146.84,165.54,168.07.LC-MS(ESI,pos,ion)m/z:394.08[M+H]。

(2) Synthesis of 2 '-hydroxy-5' -nitro- [1,1':3',1 '-terphenyl ] -3, 3' -dicarboxylic acid:

a solution of 2' -methoxy-5 ' -nitro- [1,1':3',1' -terphenyl ] -3,3 "-dicarboxylic acid (7.44g, 0.0189mol) in glacial acetic acid (50.0mL) was added to a 48% aqueous hydrobromic acid solution (50.0mL), stirred and heated at reflux for 8 hours, then cooled to room temperature and filtered to give 2' -hydroxy-5 ' -nitro- [1,1':3', 1" -terphenyl ] -3,3 "-dicarboxylic acid as a tan powder, 5.23g, 72.9% yield.

¹H-NMR(400MHz,CDCl₃)δ:4.19(s,1H),7.72(t,2H),7.85(d,2H),8.11(s,2H),8.26(d,2H),8.53(s,2H).¹³C-NMR(125MHz,CDCl₃)δ:122.22,128.57,129.72,130.59,130.94,132.94,136.72,137.77,144.25,164.92,168.07.LC-MS(ESI,pos,ion)m/z:380.07[M+H]。

(3) Synthesis of 5 '-amino-2' -hydroxy- [1,1':3',1 '-terphenyl ] -3, 3' -dicarboxylic acid:

a solution of 2 '-hydroxy-5' -nitro- [1,1':3',1 '-terphenyl ] -3, 3' -dicarboxylic acid (5.23g, 0.014mol) in ethanol (75.0mL), water (50.0mL) and a 3M sodium hydroxide solution (20.0mL) were added to a hydrogenation reaction flask at room temperature, 10% palladium on carbon (0.8g) was added thereto, nitrogen was substituted three times and hydrogen was substituted three times, the pressure was maintained at 0.3-0.5MPa, and the reaction was carried out at room temperature for 4 hours. After completion of the reaction, filtration was carried out, washing with 3M hydrochloric acid (50.0mL), followed by concentration to remove the solvent, and the crude product was slurried with 40mL of water to give 5 '-amino-2' -hydroxy- [1,1':3',1 '-terphenyl ] -3, 3' -dicarboxylic acid as a brown solid, 4.68g, 95.6%.

¹H-NMR(400MHz,CDCl₃)δ:3.39(s,2H),3.90(s,1H),6.72(s,2H),7.72(t,2H),7.85(d,2H),8.11(s,2H),8.26(d,2H).¹³C-NMR(125MHz,CDCl₃)δ:109.09,128.57,129.72,130.39,130.59,132.94,134.63,136.72,137.77,148.50,168.07.LC-MS(ESI,pos,ion)m/z:350.10[M+H]。

(4) Synthesis of Compound 1:

5 '-amino-2' -hydroxy- [1,1':3',1 "-terphenyl ] -3, 3" -dicarboxylic acid (4.68g, 0.0134mol), 2-chloro-N- (3, 4-dimethoxyphenethyl) acetamide (10.57g, 0.041mol), sodium iodide (6.00g, 0.040mol) and potassium carbonate (9.26g, 0.067mol) were suspended in dimethylformamide (30mL) and stirred at 50 ℃ for 48 hours. The solvent was concentrated off under reduced pressure, the crude product was dissolved in water (40mL), the pH was adjusted to 2.0 with hydrochloric acid, and extraction was carried out by adding dichloromethane (4X 20 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure. The crude product was purified by column chromatography (silica gel 60,400g, solvent: EA: acetone ═ 1: 1). This gave 5'- ((2- ((3, 4-dimethoxyphenethyl) amino) -2-oxoethyl) amino) -2' -hydroxy- [1,1':3',1 "-biphenyl ] -3, 3" -dicarboxylic acid (compound 1) as an off-white solid in a yield of 5.80g, 75.8%.

Example 2 Synthesis of Compound 2

5 '-amino-2' -hydroxy- [1,1':3',1 "-terphenyl ] -3, 3" -dicarboxylic acid (10g, 0.0286mol), 2-chloro-N- (4-ethoxy-3-methoxyphenethyl) acetamide (23.37g, 0.086mol), sodium iodide (12.14g, 0.081mol) and potassium carbonate (18.66g, 0.135mol) were suspended in dimethylformamide (60mL) and stirred at 50 ℃ for 48 hours. The solvent was concentrated off under reduced pressure, the crude product was dissolved in water (80mL), the pH was adjusted to 2.0 with hydrochloric acid, and extraction was carried out by adding dichloromethane (4X 40 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure. The crude product was purified by column chromatography (eluent: EA: acetone ═ 1: 1). This gave 5'- ((2- ((4-ethoxy-3-methoxyphenethyl) amino) -2-oxoethyl) amino) -2' -hydroxy- [1,1':3',1 "-biphenyl ] -3, 3" -dicarboxylic acid (compound 2) as an off-white solid in a yield of 10.94g (65.4%).

Example 3 Synthesis of Compound 3

5 '-amino-2' -hydroxy- [1,1':3',1 "-terphenyl ] -3, 3" -dicarboxylic acid (10g, 0.0286mol), 2-chloro-N- (3, 4-diethoxyphenethyl) acetamide (24.58g, 0.086mol), sodium iodide (12.14g, 0.081mol) and potassium carbonate (18.66g, 0.135mol) were suspended in dimethylformamide (60mL) and stirred at 50 ℃ for 48 hours. The solvent was concentrated off under reduced pressure, the crude product was dissolved in water (80mL), the pH was adjusted to 2.0 with hydrochloric acid, and extraction was carried out by adding dichloromethane (4X 40 mL). The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to dryness under reduced pressure. The crude product was purified by column chromatography (eluent: EA: acetone ═ 1: 1). This gave 5'- ((2- ((3, 4-diethoxyphenylethyl) amino) -2-oxoethyl) amino) -2' -hydroxy- [1,1':3',1 "-biphenyl ] -3, 3" -dicarboxylic acid (compound 3) as an off-white solid in 12.24g, 71.5% yield.

Test example 1 Effect of the Compound obtained by the present invention on proliferation of human cervical cancer cells

The antitumor cell proliferation activity of the compound obtained by the present invention was measured by the MTT method. Human cervical cancer cell lines Caski, SiHa, HeLa and C4I (Caski and SiHa are HPV16 positive cell lines, HeLa and C4I are HPV18 positive cell lines) were cultured in RPMI1640 medium containing 10% fetal bovine serum and 1% penicillin/streptomycin. The cells were cultured at 37 ℃ in a cell incubator containing 5% CO 2. Cultured cells were trypsinized and transferred to 96-well plates (2000 cells/well). After further overnight incubation, the cells were treated with the compounds obtained according to the invention at the indicated working concentrations. After 48 hours of incubation, 20uL of MTT (5mg/ml) was added dropwise to the wells. After an additional 4 hours of incubation, the mixed media in the 96-well plate was removed and then 150uL of dimethyl sulfoxide (DMSO) was added. Next, the 96-well plate was shaken for about 15 minutes, and kept at room temperature to mix the contents. The OD was then measured at a wavelength of 490 nm. The results are shown in FIG. 1. The results show that the compounds obtained by the invention all have the inhibition effect on the proliferation of human cervical cancer cells.

Test example 2 Effect of the Compound obtained by the present invention on the expression of FOXO3a protein

Cultured HeLa cells were trypsinized and transferred to 6-well plates (1X 10)⁶Cells/well). After further overnight incubation, the cells were treated with compound 1, compound 2 and compound 3 obtained according to the invention at a working concentration of 10 uM. The cells were harvested after 24 hours of treatment. After the cultured cells were harvested, they were lysed with Whole Cell Lysates (WCL), centrifuged, added with sample buffer and boiled for SDS-PAGE electrophoresis, and the electrophoresis was terminatedThe proteins were then transferred to PVDF membranes, blocked with 5% BSA and incubated overnight with primary anti-FOXO 3a and internal control protein (GAPDH)), washed with HPR-labeled secondary antibody for two hours at room temperature, and washed with chemiluminescence detection system (ECL) detection system. As shown in figure 2, the compound prepared by the invention can remarkably increase the expression of FOXO3a protein.