Application of indolepropionic acid in preparation of medicine for preventing or treating ovarian cancer

Abstract

The invention discloses an application of indolepropionic acid in preparation of a medicine for preventing or treating ovarian cancer, the medicine takes indolepropionic acid as an active ingredient, the concentration is 0.1-1000 mu M, and the dosage forms comprise tablets, capsules, granules, pills, gels, suppositories, ointments, emulsions, mixtures, suspensions, injections and solutions. Administration routes comprise oral administration, injection administration, respiratory tract inhalation administration, local administration, sublingual administration and rectal administration, and can be combined with chemotherapeutic drugs for use. It is found through experiments that IPA with different concentrations can inhibit cell growth of ovarian cancer cell lines SKOV3 and A2780, cell proliferation of macrophages RAW264.7 is remarkably promoted, and the IPA has a potential immune promotion effect and good biological safety. In addition, the IPA can enhance the sensitivity of chemotherapeutic drugs, and the IPA and the chemotherapeutic drugs are combined to be applied to treatment of ovarian cancer, so that the treatment effect of the drugs can be remarkably improved.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of indolepropionic acid in preparing a medicine for preventing or treating ovarian cancer. Background Art

[0002] Ovarian cancer (OC) is one of the most common gynecological cancers, ranking seventh in the incidence of malignant tumors and fifth in the mortality rate of female cancers worldwide. According to the World Health Organization, more than 220,000 people are diagnosed with OC and 150,000 die from OC each year. Due to the lack of obvious early symptoms of OC and the lack of effective early diagnosis strategies, 70% of patients are diagnosed with OC in the late stage.

[0003] At present, the treatment of OC is still mainly surgery and chemotherapy. Among them, platinum drugs and paclitaxel drugs are the first-line drugs for OC treatment. However, with the progress of treatment, patients' sensitivity to such drugs gradually decreases, and drug resistance develops. In addition, long-term chemotherapy may also cause serious damage or failure of multiple organs of patients, causing great physical and psychological harm to patients receiving chemotherapy. Although the treatment of OC has improved in recent years with the improvement of medication methods, the overall five-year survival rate of patients is still low. In view of this, there is an urgent need to develop a new type of OC treatment drug with high efficiency and low toxicity.

[0004] Gut microbiota-derived metabolites are the key hub connecting the gut microbiome and cancer progression, and they mainly play a role by reshaping the tumor microenvironment and regulating key signaling pathways in cancer cells and various immune cells. Tryptophan, as one of the essential amino acids for the human body, is the only amino acid in the human body that contains an indole structure. Studies have confirmed that tryptophan depletion is a prominent factor in cancer progression, which leads to the accumulation of downstream immunosuppressive metabolites, synergistically suppressing the immune system and promoting tumor growth. Tryptophan can produce indole products through the catabolism of intestinal flora, thereby regulating the body's homeostasis. Indolepropionic acid (IPA) is an important end product of indole products produced by tryptophan metabolism, which has anti-inflammatory, anti-cancer, antioxidant, and intestinal homeostasis-maintaining effects.

[0005] At present, patent document CN108125950A discloses the use of indolepropionic acid in the preparation of drugs for preventing and / or treating adverse reactions of tumor radiotherapy. Studies have found that indolepropionic acid can significantly improve intestinal damage caused by gamma-ray irradiation in mice, as manifested by a significant increase in the survival rate of experimental mice, a slight increase in body weight, and a significant decrease in intestinal inflammation levels. The experimental mice did not show abnormal reactions, indicating that indolepropionic acid can be effectively used to prevent and / or treat adverse reactions of tumor radiotherapy, has low toxicity and side effects, and is effective whether taken before or after irradiation. Patent document CN116465982A discloses the use of 3-indolepropionic acid as an endogenous biomarker in the preparation of products for diagnosing chronic kidney disease. A new marker 3-indolepropionic acid that mediates the occurrence of chronic kidney disease was screened by metabolomics and pharmacology. Experiments verified that the levels of 3-indolepropionic acid in the serum and kidney tissue of rats with 5 / 6 nephrectomy and adenine induction at different times showed an increasing trend, and the level of 3-indolepropionic acid was linearly correlated with the creatinine clearance rate. After drug treatment, the level of 3-indolepropionic acid in the serum of patients was significantly reduced, and it can be used as a new marker for diagnosing chronic kidney disease. However, in the prior art, there is no report on the use of indolepropionic acid for the prevention or treatment of ovarian cancer. Summary of the invention

[0006] Based on this, the main purpose of the present invention is to provide the use of indolepropionic acid in the preparation of drugs for preventing or treating ovarian cancer. Experiments have shown that IPA can inhibit the progression of ovarian cancer, has potential immune-promoting effects and good biosafety. In addition, IPA can also enhance the sensitivity of chemotherapeutic drugs, providing new ideas for the clinical treatment of OC.

[0007] To achieve the above object, the present invention adopts the following technical solution:

[0008] The present invention provides the use of indolepropionic acid in the preparation of a drug for preventing or treating ovarian cancer, wherein the structural formula of indolepropionic acid is

[0009] Preferably, the drug uses indolepropionic acid as an active ingredient, and its concentration is 0.1 to 1000 uM.

[0010] Preferably, the dosage form of the drug includes but is not limited to tablets, capsules, granules, pills, gels, suppositories, ointments, emulsions, mixtures, suspensions, injections and solutions.

[0011] Preferably, the administration routes of the drug include, but are not limited to, oral administration, injection, respiratory inhalation, topical administration, sublingual administration and rectal administration.

[0012] Preferably, the drug contains indolepropionic acid as an active ingredient and is used in combination with a chemotherapeutic drug.

[0013] More preferably, the chemotherapy drugs include but are not limited to platinum drugs, taxane drugs, gemcitabine, doxorubicin, bleomycin, fluorouracil, topotecan, cyclophosphamide and the like.

[0014] Preferably, indolepropionic acid is used in combination with cisplatin, wherein the concentration of indolepropionic acid is 0.1-1000uM, and the concentration of cisplatin is 0.1-50uM.

[0015] More preferably, indolepropionic acid is used in combination with cisplatin, wherein the concentration of indolepropionic acid is 500-1000 uM, and the concentration of cisplatin is 10-20 uM.

[0016] The present invention also provides a pharmaceutical composition comprising an effective dose of indolepropionic acid as an active ingredient, wherein the structural formula of indolepropionic acid is The effective dose is 0.1~1000uM.

[0017] Compared with the prior art, the present invention has the following beneficial effects: through cell experiments, the present invention found that different concentrations of IPA can inhibit the cell growth of OC cell lines SKOV3 and A2780, significantly promote the cell proliferation of macrophage RAW264.7, and has potential immune-promoting effects and good biosafety. In addition, IPA can also enhance the sensitivity of chemotherapeutic drugs, and combined with chemotherapeutic drugs for the treatment of OC can significantly improve the therapeutic effect of the drugs, and has important clinical application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The results of the effects of IPA on the proliferation of the OC cell line SKOV3 in the examples are shown.

[0019] Figure 2 The results of the effects of IPA on the proliferation of OC cell line A2780 in the examples are shown.

[0020] Figure 3 The results of the effects of IPA on the proliferation of the macrophage cell line RAW264.7 in the examples are shown.

[0021] Figure 4 The results are shown in the example below: The results show the effect of the combined use of IPA and cisplatin on the proliferation of the OC cell line SKOV3. DETAILED DESCRIPTION

[0022] The present invention is described in detail below in conjunction with specific embodiments. The following embodiments will help those skilled in the art to further understand the present invention, but are not intended to limit the present invention in any form. It should be noted that, for those of ordinary skill in the art, several changes and improvements can also be made without departing from the concept of the present invention. These all belong to the protection scope of the present invention.

[0023] Example 1

[0024] This example uses CCK8 method and cell clone formation experiment to detect the effect of IPA on the proliferation of OC cell line SKOV3 cells, as follows:

[0025] 1.1 Cell culture: SKOV3 cells were cultured in McCoy's 5A medium containing 10% fetal bovine serum, 1% penicillin and 1% streptomycin in a cell culture incubator at 37°C and 5% CO2. When the cells reached 70% to 80% confluence, they were digested and passaged with trypsin, and cells in the logarithmic growth phase and in good condition were used for subsequent experiments.

[0026] 1.2 Cell viability detection: SKOV3 cells in good growth state and in the logarithmic growth phase were selected and 1×10 4 The cells were inoculated at a density of 10 μM, 50 μM, 100 μM, 300 μM, 500 μM, and 1000 μM in a 96-well plate (5 replicate wells were set for each group). An experimental control group (containing culture medium and cells) and a blank control group (normal culture medium only, no cells) were set up. After the cells were plated, they were placed in an incubator at 37°C and 5% CO2 for 24 hours. After 24 hours, IPA was added to SKOV3 cells at drug concentrations of 10 μM, 50 μM, 100 μM, 300 μM, 500 μM, and 1000 μM. CCK-8 reagent (10 μL / well) was added after 24 hours, 48 ​​hours, and 72 hours, respectively. After incubation at 37°C for 3 hours, the absorbance was measured at a wavelength of 450 nm using an enzyme marker, and the relative cell viability value was calculated and statistically analyzed. The calculation formula was: relative cell viability (%) = (absorbance of the experimental group - absorbance of the blank group) / (absorbance of the control group - absorbance of the blank group) × 100%.

[0027] 1.3 Cell clone formation experiment: SKOV3 cells in good growth state and logarithmic growth phase were selected and inoculated in 6-well plates at a density of 700 cells / well. After plating, the cells were placed in a 37°C, 5% CO2 incubator for co-culture for 2 weeks. 24 hours after plating, IPA was added to SKOV3 cells at drug concentrations of 0μM, 50μM, 100μM, 300μM, 500μM, and 1000μM. After 72 hours, normal culture medium was changed to continue culture, and the cell medium was changed every 2 to 3 days. After 2 weeks, the culture medium was discarded, and each well of cells was washed twice with PBS, and the cells were fixed with 1mL 4% paraformaldehyde for 30min, stained with 0.1% crystal violet solution for 15min, washed with distilled water, dried, and photographed, and cell clone statistics were performed using Image J software.

[0028] Example 2

[0029] This example uses the CCK8 method to detect the effect of IPA on the proliferation of OC cell line A2780 cells, as follows:

[0030] 2.1 Cell culture: A2780 cells were cultured in DMEM medium containing 10% fetal bovine serum, 1% penicillin and 1% streptomycin in a cell culture incubator at 37°C and 5% CO2. When the cells reached 70% to 80% confluence, they were digested and passaged with trypsin, and cells in the logarithmic growth phase and in good condition were used for subsequent experiments.

[0031] 2.2 Cell viability detection: A2780 cells in good growth state and in the logarithmic growth phase were selected and 1×10 4 The cells were inoculated at a density of 10 μM, 50 μM, 100 μM, 300 μM, 500 μM, and 1000 μM in a 96-well plate (5 replicate wells were set for each group). An experimental control group (containing culture medium and cells) and a blank control group (normal culture medium only, no cells) were set up. After the cells were plated, they were placed in an incubator at 37°C and 5% CO2 for 24 hours. After 24 hours, IPA was added to A2780 cells at drug concentrations of 10 μM, 50 μM, 100 μM, 300 μM, 500 μM, and 1000 μM. CCK-8 reagent (10 μL / well) was added after 24 hours, 48 ​​hours, and 72 hours, respectively. After incubation at 37°C for 3 hours, the absorbance was measured at a wavelength of 450 nm using an enzyme marker, and the relative cell viability value was calculated and statistically analyzed. The calculation formula was: relative cell viability (%) = (absorbance of the experimental group - absorbance of the blank group) / (absorbance of the control group - absorbance of the blank group) × 100%.

[0032] Example 3

[0033] In this example, the CCK8 method was used to detect the effect of IPA on the proliferation of macrophage RAW264.7, as follows:

[0034] 3.1 Cell culture: RAW264.7 cells were cultured in DMEM medium containing 10% fetal bovine serum, 1% penicillin and 1% streptomycin in a cell culture incubator at 37°C and 5% CO2. When the cells reached 70% to 80% confluence, they were passaged and cells in the logarithmic growth phase and in good condition were used for subsequent experiments.

[0035] 3.2 Cell viability detection: RAW264.7 cells in good growth state and logarithmic growth phase were selected and 1×10 4The cells were inoculated at a density of 10 μM, 50 μM, 100 μM, 300 μM, 500 μM, and 1000 μM in a 96-well plate (5 replicate wells were set for each group). An experimental control group (containing culture medium and cells) and a blank control group (normal culture medium only, no cells) were set up. After the cells were plated, they were placed in an incubator at 37°C and 5% CO2 for 24 hours. After 24 hours, IPA was added to RAW264.7 cells at drug concentrations of 10 μM, 50 μM, 100 μM, 300 μM, 500 μM, and 1000 μM. CCK-8 reagent (10 μL / well) was added after 24 hours, 48 ​​hours, and 72 hours, respectively. After incubation at 37°C for 3 hours, the absorbance was measured at a wavelength of 450 nm using an enzyme marker, and the relative cell viability value was calculated and statistically analyzed. The calculation formula was: relative cell viability (%) = (absorbance of the experimental group - absorbance of the blank group) / (absorbance of the control group - absorbance of the blank group) × 100%.

[0036] Example 4

[0037] This example uses CCK8 method to detect the effect of IPA combined with cisplatin on the proliferation of OC cell line SKOV3 cells, as follows:

[0038] 4.1 Cell culture: Same as 1.1 in Example 1.

[0039] 4.2 Cell viability detection: SKOV3 cells in good growth state and in the logarithmic growth phase were selected and 1×10 4 The cells were inoculated at a density of 1 μM / well in a 96-well plate (5 replicate wells were set for each group). A cisplatin monotherapy group (DDP) and an IPA and cisplatin combination group (DDP+IPA) were set up. After cell plating, the cells were placed in an incubator at 37°C and 5% CO2 for 24 hours. After 24 hours, cisplatin was added to SKOV3 cells in the DDP group at drug concentrations of 1μM, 5μM, 10μM, 20μM, and 40μM. In the DDP+IPA group, 1000μM IPA was added to each well while cisplatin was added at the above drug concentrations. CCK-8 reagent (10μL / well) was added 48 hours later. After incubation at 37°C for 3 hours, the absorbance was measured at a wavelength of 450nm using an enzyme reader. The relative cell viability value was calculated and statistical analysis was performed. The calculation formula was: relative cell viability (%) = (absorbance of the experimental group - absorbance of the blank group) / (absorbance of the control group - absorbance of the blank group) × 100%.

[0040] GraphPad Prism software was used for data analysis. One-way ANOVA was used to analyze the differences among the groups. The data were expressed as mean ± standard deviation (SD). When p < 0.05, the difference was considered statistically significant. Figures 1 to 4 shown.

[0041] like Figure 1As shown in the results of CCK8, IPA has an inhibitory effect on the cell proliferation of SKOV3 cells in a time-dependent and dose-dependent manner. As the intervention time increases, the effective dose concentration of IPA gradually decreases. After 72 hours, all dose groups can significantly inhibit the proliferation of SKOV3 cells ( Figure 1 A). The results of the cell clone formation experiment also showed that high doses of IPA could significantly reduce the formation of SKOV3 cell clones, confirming that IPA has a significant inhibitory effect on the cell proliferation of SKOV3 cells ( Figure 1 B).

[0042] like Figure 2 As shown, IPA had a significant inhibitory effect on the proliferation of A2780 cells, but did not show significant time-dependency and dose-dependency.

[0043] like Figure 3 As shown in the figure, IPA has a significant promoting effect on RAW264.7 cell proliferation in a short period of time (24h), indicating its great potential in promoting immunity. As the intervention time increases, the promoting effect is no longer significant, but long-term, high-dose intervention of IPA does not show a significant inhibitory effect on RAW264.7 cell proliferation, which indirectly reflects its good biosafety.

[0044] like Figure 4 As shown, the IC50 value of cisplatin acting alone on SKOV3 cells for 48 hours was 17.92 μM, and the IC50 value of cisplatin acting in combination with 1000 μM IPA for 48 hours was 12.72 μM. IPA intervention can increase the sensitivity of SKOV3 cells to cisplatin.

[0045] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. Use of indolepropionic acid in the preparation of a drug for preventing or treating ovarian cancer, wherein the structural formula of indolepropionic acid is 2. The use according to claim 1, characterized in that: The drug uses indolepropionic acid as an active ingredient, and its concentration is 0.1 to 1000 uM.

3. The use according to claim 1, characterized in that: The dosage forms of the drug include tablets, capsules, granules, pills, gels, suppositories, ointments, emulsions, mixtures, suspensions, injections and solutions.

4. The use according to claim 1, characterized in that: The administration routes of the drug include oral administration, injection, respiratory inhalation, topical administration, sublingual administration and rectal administration.

5. The use according to claim 1, characterized in that: The drug uses indolepropionic acid as an active ingredient and is used in combination with a chemotherapy drug.

6. The use according to claim 5, characterized in that: The chemotherapy drug is selected from one or more of platinum drugs, taxane drugs, gemcitabine, doxorubicin, bleomycin, fluorouracil, topotecan, and cyclophosphamide.

7. The use according to claim 5, characterized in that: Indolepropionic acid is used in combination with cisplatin, wherein the concentration of indolepropionic acid is 0.1-1000uM, and the concentration of cisplatin is 0.1-50uM.

8. The use according to claim 7, characterized in that: Indolepropionic acid is used in combination with cisplatin, wherein the concentration of indolepropionic acid is 500-1000uM, and the concentration of cisplatin is 10-20uM.

9. A pharmaceutical composition, characterized in that It contains an effective dose of indolepropionic acid as an active ingredient, wherein the structural formula of indolepropionic acid is The effective dose is 0.1~1000uM.

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