Use of flavonoid compound and drug for preventing and / or treating familial intestinal polyposis

Abstract

A use of a flavonoid compound and a drug for preventing and / or treating familial intestinal polyposis. In the present invention, it is found for the first time by experiments that the compound as shown in formula I can correct the disorder of β-catenin accumulation in cell nuclei caused by APC gene mutation to promote the transfer of β-catenin to cell membranes, can promote the restoration of the normal morphology of intestinal villi with dysplasia and deformity, and can decrease the number of intestinal polyps caused by APC mutation, thereby achieving the effect of preventing and treating familial intestinal polyposis.

Description

Use of flavonoid compound and medicine for preventing and / or treating familial intestinal polyps Technical Field

[0001] The present invention belongs to the field of biotechnology, and in particular relates to the use of a flavonoid compound and a medicine for preventing and / or treating familial intestinal polyps. Background Art

[0002] Familial Adenomatous Polyposis (FAP) is an inherited disorder that is usually caused by mutations in the APC gene. Polyps are small bumps or masses in the intestine that are usually benign, but they can develop into cancer in people with FAP. In addition, people with FAP may also develop polyps in other organs, most commonly the duodenum. Polyps in the duodenum can also develop into cancer. Polyps that develop in the colon are usually monitored and removed through regular colonoscopies that continue throughout life, usually starting in adolescence. To reduce the risk of cancer, colectomy, the surgical removal of the entire colon, is a common treatment for FAP.

[0003] β-catenin is a multifunctional protein that plays a key role in intracellular signaling and gene transcription regulation. It is an important structural protein involved in forming intercellular junctions and interacts with a variety of proteins to regulate cell growth, differentiation, migration, and apoptosis. Under normal physiological conditions, β-catenin levels in cells are strictly regulated. However, under certain abnormalities, such as gene mutations or abnormal signaling, β-catenin levels in cells can change, leading to uncontrolled cell growth and ultimately tumorigenesis. Therefore, the study of β-catenin is of great significance for understanding cell biology and tumorigenesis. Currently, there are no reports on how to regulate β-catenin to achieve the treatment of FAP.

[0004] Some medications, such as nonsteroidal anti-inflammatory drugs (NSAIDs) or selective cyclooxygenase-2 inhibitors (such as Celecoxib), may be helpful in the treatment of FAP. These drugs can slow the growth of polyps but are not a cure. Therefore, there is an urgent need to develop new drugs to treat FAP.

[0005] The small molecule compound YS434 has the molecular formula:

[0006] And the small molecule compound YS392, the molecular formula is:

[0007] According to reports (CN201810380559.X: Flavonoids for the prevention and treatment of colorectal cancer, and CN202210977121.6: A novel use of a flavonoid for the treatment of ulcerative colitis), these compounds have the potential to prevent and treat colorectal cancer and ulcerative colitis. However, there are no reports of the efficacy of these small molecule compounds in treating familial intestinal polyps. Summary of the Invention

[0008] In view of the problems of the prior art, the present invention provides a use of a flavonoid compound and a medicine for preventing and / or treating familial intestinal polyps.

[0009] The use of a compound represented by Formula I, or a derivative thereof, or a pharmaceutically acceptable salt thereof, or a crystal form thereof, or a stereoisomer thereof, or an optical isomer thereof, in the preparation of a medicament for preventing and / or treating familial intestinal polyposis, is characterized in that the structural formula of the compound represented by Formula I is as follows:

[0010] Wherein, R is selected from C1-C 10 alkoxy or amino groups.

[0011] Preferably, the R is selected from a C2 alkoxy group or an amino group.

[0012] Preferably, the compound represented by formula I is:

[0013] Preferably, the drug is used to improve the accumulation of β-catenin in the cell nucleus and promote the transfer of β-catenin to the cell membrane;

[0014] And / or, the drug is used to promote the restoration of normal morphology of abnormally developed and / or deformed intestinal villi.

[0015] Preferably, the amino acid sequence of β-catenin is shown as SEQ ID NO.1.

[0016] Preferably, the drug is in the form of a pill, tablet, solution, inhaler or spray.

[0017] The present invention also provides a drug for preventing and / or treating familial intestinal polyps, which is prepared by using the compound represented by Formula I, or its derivative, or its pharmaceutically acceptable salt, or its crystal form, or its stereoisomer, or its optical isomer as the active ingredient, and adding pharmaceutically acceptable excipients or auxiliary ingredients;

[0018] The structural formula of the compound shown in Formula I is as follows:

[0019] Wherein, R is selected from C1-C 10 alkoxy or amino groups.

[0020] Preferably, the R is selected from a C2 alkoxy group or an amino group.

[0021] Preferably, the compound represented by formula I is:

[0022] Preferably, the drug is used to improve the accumulation of β-catenin in the cell nucleus and promote the transfer of β-catenin to the cell membrane;

[0023] And / or, the drug is used to promote the restoration of normal morphology of abnormally developed and / or deformed intestinal villi.

[0024] Preferably, the drug is in the form of a pill, tablet, solution, inhaler or spray.

[0025] In the present invention, the amino acid sequence of β-catenin (SEQ ID NO. 1) is as follows:

[0026] The present invention provides a new use for the flavonoid compounds represented by Formula I. Experimental findings indicate that these compounds can correct the accumulation of β-catenin in the cell nucleus caused by APC gene mutations, promote the transfer of β-catenin to the cell membrane, and promote the restoration of dysplastic and deformed intestinal villi to normal morphology. They can also reduce the number of intestinal polyps caused by APC mutations, thereby preventing and treating familial intestinal polyps. This invention provides a new option for the development of drugs for familial intestinal polyps and has promising application prospects.

[0027] Obviously, based on the above contents of the present invention, according to common technical knowledge and customary means in this field, without departing from the above basic technical ideas of the present invention, other various forms of modifications, replacements or changes can be made.

[0028] The following further describes the above content of the present invention in detail through specific embodiments in the form of examples. However, this should not be construed as limiting the scope of the above subject matter of the present invention to the following examples. All technologies implemented based on the above content of the present invention fall within the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 shows the experimental results of YS434 preventing β-catenin from entering the cell nucleus. A is a representative detection result of Western-blot analysis of cytoplasmic and nuclear proteins extracted from cells after 48 hours of YS434 treatment; B is the statistical analysis result of the Western-blot test results (*: P<0.05, **: P<0.001).

[0030] Figure 2 shows the experimental results of YS434 promoting the transfer of β-catenin to the cell membrane. A is a representative detection result of a Western-blot experiment in which cell membrane proteins were extracted after 48 hours of YS434 treatment; B is the statistical analysis result of the Western-blot experiment (*: P < 0.05, ***: P < 0.0001).

[0031] Figure 3 shows the experimental results of YS434 promoting the translocation of β-catenin to the cell membrane. A is the result of a landmark experiment using immunofluorescence staining using anti-E-cadherin and anti-β-catenin antibodies after 48 hours of YS434 treatment; B is the result of colocalization analysis of antibody-labeled E-cadherin and β-catenin (shown as Pearson correlation coefficient of localization, **: P<0.001).

[0032] Figure 4 shows the effect of YS434 on APC min / + The experimental results of the reduction in the number of intestinal polyps in mice are shown in Figure 1, where A is the statistical result of the number of polyp nodules in the small intestine of the control group (given corn oil) mice and the treatment group (given YS434) mice one month after administration; B is the statistical result of the number of polyp nodules in the colorectal area of ​​the control group and the treatment group mice one month after administration (*: P<0.05).

[0033] Figure 5 shows the effect of YS434 on APC min / + Figure 1 shows the experimental results of mouse intestinal villus morphology returning from abnormality to normal, where A is the HE staining result of paraffin sections of intestinal tissue of 5-month-old wild-type mice (i.e., healthy mice without APC gene mutation); B is the HE staining result of paraffin sections of intestinal tissue of 4-month-old APC min / + HE staining results of paraffin sections of intestinal tissue of mice fed corn oil for 1 month; C is APC at 4 months old min / + HE staining results of paraffin sections of intestinal tissue of mice 1 month after administration of YS434. DETAILED DESCRIPTION

[0034] In the following examples and experimental examples, reagents and raw materials not specifically described are all commercially available.

[0035] Example 1 Therapeutic Effect of Small Molecule Compound YS434 on Familial Intestinal Polyps

[0036] It should be noted that normal human intestinal epithelial cells are difficult to culture in vitro, and their behavior in vitro, especially in 2D culture, differs significantly from that of in vivo intestinal epithelial cells, making them generally unsuitable for use as a model for epithelial cell function studies. Caco-2 cells are morphologically similar to normal human small intestinal epithelial cells, sharing the same cell polarity and tight junctions, and exhibit similar molecular characteristics to normal intestinal epithelial cells. As a general model, they are widely used to study the physiological functions of intestinal epithelial cells. However, primary human colon cancer cells, due to their potential for differentiation, are an ideal model for this application's research related to cell differentiation induction.

[0037] 1. YS434 prevents β-catenin from entering the cell nucleus in cell experiments

[0038] 1. Experimental methods

[0039] Human colon cell line Caco-2 was cultured in DMEM + 10% fetal bovine serum. Different concentrations of YS434 were added to the culture medium. After treatment for 48 hours, the cells were collected and nuclear and cytoplasmic proteins were extracted using the Pierce NE-PER nuclear and cytoplasmic protein extraction kit produced by Thermo-Fisher, respectively, for Western-blot analysis.

[0040] 2. Experimental results

[0041] The results are shown in Figure 1. As the concentration of YS434 increases, the β-catenin content in the cell nucleus decreases, indicating that YS434 can significantly inhibit the entry of β-catenin into the cell nucleus.

[0042] 2. YS434 promotes β-catenin translocation to the cell membrane in cell experiments

[0043] 1. Experimental methods

[0044] Method 1: Primary human colon cancer cells were cultured in DMEM + 10% fetal bovine serum medium. Different concentrations of YS434 were added to the medium. After treating the cells for 48 hours, the cells were collected and the cell membrane proteins were extracted using the Thermo-Fisher Mem-PER Plus membrane protein extraction kit for Western blot detection.

[0045] Method 2: Primary human colon cancer cells were seeded in a culture dish with a matrigel base and cultured in DMEM + 10% fetal bovine serum. YS434 (2 μg / ml) was added to the culture medium. After treatment for 48 hours, β-catenin and cell membrane marker E-cadherin were detected by immunofluorescence staining.

[0046] 2. Experimental results

[0047] The results of Method 1 are shown in Figure 2. After YS434 treatment, there was a dose-dependent increase in β-catenin on the cell membrane (positive for the internal reference membrane protein ATPase, and negative for the nuclear protein lamin and the cytoplasmic protein GAPDH, indicating that the sample was a membrane protein), indicating that YS434 treatment specifically promotes β-catenin localization on the cell membrane. The results of Method 2 are shown in Figure 3. DAPI staining revealed a decrease in β-catenin staining in the nucleus, indicating that YS434 treatment inhibited β-catenin entry into the nucleus. Concurrently, β-catenin increased near the cell membrane, as indicated by E-cadherin staining, and statistical analysis confirmed an increase in the colocalization coefficient between β-catenin and the membrane protein E-cadherin, suggesting that YS434 treatment caused β-catenin to accumulate at the cell membrane.

[0048] 3. YS434 leads to APC min / + The number of intestinal polyps in mice was significantly reduced

[0049] 1. Experimental methods

[0050] Take 4-month-old APC min / + Mice (purchased from Jicui Pharmaceuticals) were gavaged with YS434 (dissolved in corn oil) at 20 mg / kg body weight every five days for one month. A control group received corn oil alone. One month later, the intestines of the control and YS434-treated mice were harvested and the number of nodules was counted.

[0051] 2. Experimental results

[0052] The results are shown in Figure 4. After YS434 treatment, the number of intestinal polyp nodules was significantly lower than that in the control group, indicating that YS434 effectively inhibited and reversed the occurrence of intestinal polyps caused by APC mutation.

[0053] 4. YS434 leads to APC min / + The morphology of mouse intestinal villi returned from abnormality to normal

[0054] 1. Experimental methods

[0055] Take 4-month-old APC min / + Mice were administered YS434 (dissolved in corn oil) by gavage at 20 mg / kg body weight every five days for one month. A control group received corn oil alone. After one month, intestinal tissues from the control and YS434-treated mice were prepared and paraffin sections were prepared for H&E staining.

[0056] 2. Experimental results

[0057] As shown in Figure 5, APC min / +The villi of the small intestine of mice were swollen, twisted and deformed, and YS434 treatment restored the morphology of the villi to normal.

[0058] The experimental data above demonstrate that the compounds provided by this invention can correct the β-catenin accumulation disorder in the cell nucleus caused by APC gene mutations, promote β-catenin translocation to the cell membrane, and restore dysplastic and deformed intestinal villi to normal morphology. They can also reduce the number of intestinal polyps caused by APC mutations, thereby preventing and treating familial intestinal polyps. This provides more options for the medical treatment of familial intestinal polyps and has promising application prospects.

Claims

1. Use of a compound of formula I, or a derivative thereof, or a pharmaceutically acceptable salt thereof, or a crystal form thereof, or a stereoisomer thereof, or an optical isomer thereof in the preparation of a drug for preventing and / or treating familial intestinal polyps, characterized in that: The structural formula of the compound shown in Formula I is as follows: Wherein, R is selected from C1-C 10 of an alkoxy or amine group.

2. The use according to claim 1, characterized in that: The R is selected from a C2 alkoxy group or an amine group.

3. The use according to claim 2, characterized in that: The compound shown in formula I is:

4. The use according to claim 1, characterized in that: The drug is used to improve the accumulation of β-catenin in the cell nucleus and promote the transfer of β-catenin to the cell membrane; And / or, the drug is used to promote the restoration of normal morphology of abnormally developed and / or deformed intestinal villi.

5. The use according to claim 4, characterized in that: The amino acid sequence of β-catenin is shown in SEQ ID NO.

1.

6. The use according to claim 1, characterized in that: The drug is in the form of pills, tablets, solutions, inhalers or sprays.

7. A drug for preventing and / or treating familial intestinal polyps, characterized in that: It is prepared by using the compound shown in formula I, or its derivative, or its pharmaceutically acceptable salt, or its crystal form, or its stereoisomer, or its optical isomer as the active ingredient, and adding pharmaceutically acceptable excipients or auxiliary ingredients; The structural formula of the compound shown in Formula I is as follows: Wherein, R is selected from C1-C 10 of an alkoxy or amine group.

8. The drug according to claim 7, characterized in that: The R is selected from a C2 alkoxy group or an amine group.

9. The drug according to claim 8, characterized in that: The compound shown in formula I is:

10. The drug according to claim 6, characterized in that: The drug is in the form of pills, tablets, solutions, inhalers or sprays.

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