Application of tetrahydropalmatine in preparation of chemotherapy sensitizing drug and chemotherapy sensitizing drug

Abstract

The invention relates to the field of medicines, and discloses an application of tetrahydropalmatine in preparation of a chemotherapy sensitizing medicine and the chemotherapy sensitizing medicine. The tetrahydropalmatine disclosed by the invention is applied to the chemotherapy sensitizing medicine, and can inhibit cell activity, inhibit cell proliferation, promote cell apoptosis and inhibit cell migration. The chemotherapy sensitizing drug is used for increasing the sensitivity of malignant tumors to chemotherapy drugs, and after tetrahydropalmatine and paclitaxel are combined for use, the survival rate of paclitaxel to ovarian cancer drug-resistant cells A2780 / Taxol is only 30.38%. Tetrahydropalmatine and paclitaxel are combined to weaken the anti-apoptosis ability and enhance the apoptosis promoting ability by regulating the Bcl-2 / Bax ratio, and conduction of an apoptosis signal channel is promoted by activating a Caspase cascade reaction. The combined use of the tetrahydropalmatine and the paclitaxel can almost completely inhibit the migration of A2780 / Taxol cells within 48 hours.

Description

Technical Field

[0001] The present invention relates to the field of medicines, and more specifically to an application of tetrahydropalmatine in the preparation of a chemotherapy sensitizing drug and the chemotherapy sensitizing drug. Background Art

[0002] Ovarian cancer is a common malignant tumor of the female reproductive system and one of the leading causes of cancer death in women. According to global cancer statistics, the incidence of ovarian cancer in women ranks eighth, accounting for 3.7% of global cancer cases and 4.7% of cancer deaths in 2020. The early symptoms of ovarian cancer are not obvious and are often ignored or misdiagnosed. Therefore, more than two-thirds of patients are in the advanced stage when diagnosed, which seriously affects the treatment effect. Although treatment methods such as surgery, radiotherapy and chemotherapy can improve the survival of patients to a certain extent, the treatment effect is still not ideal due to the large toxic side effects of chemotherapy drugs during the treatment process, and the heterogeneity and drug resistance of tumor cells limit the efficacy.

[0003] Currently, chemotherapy based on platinum drugs and paclitaxel is one of the main methods for treating ovarian cancer. In particular, paclitaxel, as a commonly used chemotherapy drug, can effectively relieve the symptoms of ovarian cancer patients. However, the clinical application of paclitaxel is restricted by its greater toxicity and drug resistance. Studies have found that the drug resistance of ovarian cancer cells gradually develops during the treatment process, resulting in a significant decrease in the efficacy of paclitaxel. Therefore, the development of new chemotherapy sensitizers to increase the sensitivity of chemotherapy drugs and reduce the side effects of drugs has become an important research direction in the field of tumor treatment.

[0004] Chemosensitizers can enhance the sensitivity of tumor cells to chemotherapy drugs without increasing the toxicity of chemotherapy drugs, thereby improving the treatment effect. By using them in combination with chemotherapy drugs, sensitizers are expected to reduce patients' need for high-dose chemotherapy drugs and reduce their side effects, while overcoming the drug resistance of tumor cells and delaying or reversing the occurrence of drug resistance. In particular, paclitaxel is an important drug for ovarian cancer chemotherapy. If a sensitizer that matches it can be found, it is expected to significantly improve its efficacy, effectively reduce the dosage of chemotherapy drugs, and reduce the side effects of chemotherapy in patients. It has important practical significance for the treatment of ovarian cancer and reversing / delaying the occurrence of ovarian cancer resistance.

[0005] Tetrahydropalmatine (also known as tetrahydropalmatine) is an important biologically active ingredient derived from the traditional Chinese medicine Corydalis yanhusuo, which has multiple physiological activities such as sedation, hypnosis, analgesia, anti-arrhythmia and anti-tumor. Prior art 201010262945.2 discloses the use of tetrahydropalmatine in the preparation of drugs for inhibiting the proliferation of eukaryotic tumor cells. Although tetrahydropalmatine has shown certain research potential in the field of anti-tumor, there is currently no relevant research report on the chemosensitization function of tetrahydropalmatine. Summary of the invention

[0006] The present invention is to overcome the problem that the prior art described above has no application of tetrahydropalmatine in enhancing the sensitivity of chemotherapy drugs, and provides an application of tetrahydropalmatine in the preparation of chemotherapy sensitization drugs;

[0007] Another object of the present invention is to provide a chemotherapy sensitization drug.

[0008] In order to solve the above technical problems, the technical solution of the present invention is as follows:

[0009] A use of tetrahydropalmatine in the preparation of a chemotherapy sensitizing drug, for preparing a chemotherapy sensitizing drug; the structural formula of the tetrahydropalmatine is:

[0010]

[0011] Furthermore, the chemotherapy sensitizing drug is used to inhibit cell activity, inhibit cell proliferation, promote cell apoptosis, and inhibit cell migration.

[0012] Furthermore, the cells include cells that are resistant to chemotherapeutic drugs.

[0013] Furthermore, the chemotherapy sensitizer is used to increase the sensitivity of malignant tumors to chemotherapy drugs.

[0014] Furthermore, the malignant tumor includes a malignant tumor of the female reproductive system.

[0015] Preferably, the female reproductive system malignancy includes ovarian cancer.

[0016] Furthermore, the chemotherapy drug includes paclitaxel.

[0017] A chemotherapy sensitizing drug, comprising at least one of tetrahydropalmatine, a tetrahydropalmatine derivative and a pharmaceutically acceptable salt thereof; the structural formula of the tetrahydropalmatine is:

[0018]

[0019] Furthermore, the chemotherapy sensitizing drug also includes a chemotherapy drug component.

[0020] Furthermore, the chemotherapy drug component includes paclitaxel.

[0021] Furthermore, the molar concentration ratio of tetrahydropalmatine to paclitaxel is 0.5:1 to 1600:1.

[0022] Preferably, further, the drug concentration ratio of tetrahydropalmatine and paclitaxel is 0.5:1 to 1600:1.

[0023] Preferably, the molar concentration ratio of tetrahydropalmatine to paclitaxel is 6.25-512:1.

[0024] Preferably, the molar concentration ratio of tetrahydropalmatine to paclitaxel is 12.5-400:1.

[0025] Preferably, the molar concentration ratio of tetrahydropalmatine to paclitaxel is 400:1.

[0026] Preferably, the molar concentration of tetrahydropalmatine is 5 to 50 μM.

[0027] Tetrahydropalmatine is a bioactive molecule derived from traditional Chinese medicine. Previous studies have mainly focused on its traditional pharmacological effects such as anti-inflammatory, analgesic, and anti-arrhythmic. The present invention proposes for the first time and systematically verifies the application of Tetrahydropalmatine in the field of chemotherapy sensitization, especially its sensitization effect on paclitaxel-resistant ovarian cancer cells. This innovative application fills the gap in the research field of Tetrahydropalmatine's chemotherapy sensitization function. Tetrahydropalmatine can show a better sensitization effect at low concentrations. When used in combination with paclitaxel, the survival rate of resistant cells is significantly reduced. This synergistic effect can not only improve the efficacy of chemotherapy drugs, but also significantly reduce the effective dosage of paclitaxel, thereby reducing the toxic side effects caused by high-dose chemotherapy drugs.

[0028] Compared with the prior art, the technical solution of the present invention has the following beneficial effects:

[0029] 1. Improve the sensitivity of chemotherapy drugs. When Tetrahydropalmatine is used in combination with paclitaxel, it significantly enhances the sensitivity of paclitaxel to ovarian cancer resistant cells A2780 / Taxol. When the concentration of Tetrahydropalmatine is 5μM, when it is used in combination with 10μM paclitaxel, the cell viability is significantly reduced, and the cell survival rate is the lowest, only 30.38%. Tetrahydropalmatine can effectively improve the anti-cancer effect of paclitaxel and significantly increase the chemotherapy sensitivity of paclitaxel.

[0030] 2. Promote cell apoptosis. When corydalis ethyl is used in combination with paclitaxel, it can significantly promote the apoptosis of ovarian cancer resistant cells A2780 / Taxol. By regulating the Bcl-2 / Bax ratio, the anti-apoptotic ability is weakened and the pro-apoptotic ability is enhanced. It can also activate the Caspase cascade reaction (Cleaved-PARP increases significantly) and promote the conduction of the apoptotic signaling pathway. After corydalis ethyl is treated with paclitaxel, the resistant cells shrink, fall off or break over a large area, and the density is significantly reduced. When paclitaxel or corydalis ethyl is used alone, it hardly causes cell apoptosis.

[0031] 3. Inhibit cell migration. When tetrahydropalmatine was used in combination with paclitaxel, the migration of A2780 / Taxol cells was almost completely inhibited within 48 hours, while the inhibitory effect of paclitaxel or tetrahydropalmatine alone on cell migration was limited. This indicates that tetrahydropalmatine significantly enhances the anti-migratory effect of paclitaxel.

[0032] 4. Reduce toxicity and enhance efficacy. Tetrahydropalmatine has low toxicity when used alone, but when used in combination with paclitaxel, it significantly enhances the inhibitory effect of paclitaxel on cancer cells without increasing the dose of paclitaxel. This not only reduces the dose of chemotherapy drugs, but also helps reduce the toxic side effects of high-dose paclitaxel and improves patient tolerance. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 Schematic diagram of the inhibitory effect of tetrahydropalmatine combined with paclitaxel (molar concentration ratio: 0.5-512:1) on the activity of paclitaxel-resistant ovarian cancer cells A2780 / Taxol, # indicates P<0.05, ## indicates P<0.01, and ### indicates P<0.001;

[0034] Figure 2 Schematic diagram of the inhibitory effect of tetrahydropalmatine combined with paclitaxel (molar concentration ratio: 6.25-1600:1) on the activity of paclitaxel-resistant ovarian cancer cells A2780 / Taxol, # indicates P<0.05, ## indicates P<0.01, and ### indicates P<0.001;

[0035] Figure 3 Schematic diagram of the inhibitory effect of tetrahydropalmatine combined with paclitaxel (molar concentration ratio: 100-400:1) on the activity of paclitaxel-resistant ovarian cancer cells A2780 / Taxol, # indicates P<0.05, ## indicates P<0.01, and ### indicates P<0.001;

[0036] Figure 4 Schematic diagram of the inhibitory effect of tetrahydropalmatine on the activity of paclitaxel-resistant ovarian cancer A2780 / Taxol cells;

[0037] Figure 5 The effect of tetrahydropalmatine combined with paclitaxel (substance concentration ratio 400:1) on paclitaxel-resistant ovarian cancer cells A2780 / Taxol; A. Cell pictures of each group after drug intervention; B. Cell apoptosis of each group after drug intervention;

[0038] Figure 6 This is a graph showing the inhibitory effect of tetrahydropalmatine combined with paclitaxel (at a concentration ratio of 400:1) on the proliferation of paclitaxel-resistant ovarian cancer cells A2780 / Taxol;

[0039] Figure 7 To investigate the effect of tetrahydropalmatine combined with paclitaxel (at a concentration ratio of 400:1) on the expression of PARP, Cleaved-PARP, Bax, Bcl-2 and other proteins in paclitaxel-resistant ovarian cancer cells A2780 / Taxol.

[0040] Figure 8 This is a graph showing the inhibitory effect of corydalis ethyl on the migration of paclitaxel-resistant ovarian cancer cells A2780 / Taxol when used in combination (molar concentration ratio 400:1). DETAILED DESCRIPTION

[0041] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0042] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0043] Example 1

[0044] Ovarian cancer paclitaxel-resistant cells A2780 / Taxol were seeded in 96-well plates, with 5.5×10 3 cells, and the cell culture medium was aspirated after 24 hours of culture, and a negative control group and a drug-treated group were set up. In the drug-treated group, 5 μM tetrahydropalmatine was added to each well, and 0.00975, 0.0390620.15625, 0.625, 2.5, and 10 μM paclitaxel were used in combination (the molar concentration ratio of tetrahydropalmatine to paclitaxel: 0.5-512:1), and only the corresponding concentration of paclitaxel was added to the negative control group, and cultured for 48 hours. This experiment was repeated three times.

[0045] Example 2

[0046] Ovarian cancer paclitaxel-resistant cells A2780 / Taxol were seeded in 96-well plates, with 5.5×10 3 cells, and the cell culture medium was aspirated after 24 hours of culture, and a negative control group and a drug-treated group were set up. In the drug-treated group, 50 μM tetrahydropalmatine was added to each well, and 0.03125, 0.125, 0.5, 2, and 8 μM paclitaxel were used in combination (the molar concentration ratio of tetrahydropalmatine to paclitaxel was 6.25-1600:1), and only the corresponding concentration of paclitaxel was added to the negative control group, and cultured for 48 hours. This experiment was repeated three times.

[0047] Example 3

[0048] Ovarian cancer paclitaxel-resistant cells A2780 / Taxol were seeded in 96-well plates, with 5.5×10 3cells, and the cell culture medium was aspirated after 24 hours of culture, and a negative control group and a drug-treated group were set up. Paclitaxel 125nM and tetrahydropalmatine 12.5, 25, and 50μM were added to each well of the drug-treated group (the molar concentration ratio of tetrahydropalmatine to paclitaxel: 100-400:1), and tetrahydropalmatine of the corresponding concentration was added to the negative control group only, and cultured for 48 hours. This experiment was repeated three times.

[0049] Example 4

[0050] Ovarian cancer paclitaxel-resistant cells A2780 / Taxol were seeded in 6-well plates, with 2×10 5 cells, and the cell culture medium was aspirated after culturing for 24 hours. Paclitaxel and tetrahydropalmatine (125 nM paclitaxel and 50 μM tetrahydropalmatine) with a molar concentration ratio of 1:400 were added to the paclitaxel+tetrahydropalmatine group, while paclitaxel 125 nM was added to the paclitaxel group alone, and tetrahydropalmatine 50 μM was added to the tetrahydropalmatine group alone. The cells were returned to the incubator and cultured for 48 hours.

[0051] Example 5

[0052] Ovarian cancer paclitaxel-resistant cells A2780 / Taxol were seeded in 6-well plates, with 4.5×10 5 cells, and cultured until the cell confluence reached 80%, then scratched the cells with a 10 μL plastic pipette tip, washed away the floating cells with PBS, and added paclitaxel and corydalis with a molar concentration ratio of 1:400 (paclitaxel 125 nM, corydalis 50 μM) to the paclitaxel+corydalis group, while no paclitaxel and corydalis were added to the normal group. Paclitaxel 125 nM was added to the paclitaxel group alone, and corydalis 50 μM was added to the corydalis group alone. The cells were returned to the incubator and cultured for 48 h.

[0053] Example 6

[0054] Ovarian cancer paclitaxel-resistant cells A2780 / Taxol were seeded in 6-well plates, with 5×10 2 After the cells were cultured overnight and adhered to the wall, paclitaxel and tetrahydropalmatine (paclitaxel 125nM, tetrahydropalmatine 50μM; no addition was added to the control group) were added at a substance concentration ratio of 1:400, and returned to the incubator for static culture for 1 to 2 weeks, during which the culture medium was replaced every 2 days. The culture was terminated when visible clones were observed.

[0055] Comparative Example 1

[0056] Ovarian cancer paclitaxel-resistant cells A2780 / Taxol were seeded in 96-well plates, with 5.5×10 3cells, cultured for 24 hours, aspirated the cell culture medium, added a full culture medium containing tetrahydropalmatine (1.5625, 3.125, 6.25, 12.5, 25, 50 μM) and DMSO (negative control) to each well, continued to culture for 48 hours, discarded the supernatant, added a full culture medium containing MTT (5 mg / mL), cultured for 3 hours, aspirated the supernatant, added 100 μL of DMSO to each well, oscillated for 10 minutes, measured the absorbance at 490 nm (A value), and the blank control was the B value. Calculate the cell survival rate (experimental group A value / control group B value*100%), and repeat this experiment three times.

[0057] Detection Methods

[0058] 1. Cell viability detection

[0059] The supernatant of the obtained cell culture fluid was discarded, and a full culture medium containing MTT (5 mg / mL) was added. After culturing for 3 hours, the supernatant was aspirated, 100 μL of DMSO was added to each well, and the cells were shaken for 10 minutes. The absorbance (A value) was measured at 490 nm, and the blank control was the B value. The cell survival rate was calculated (experimental group A value / control group B value × 100%).

[0060] 2. Cell apoptosis detection

[0061] The supernatant of the obtained cell culture fluid was discarded, the cells were collected by centrifugation, and washed with pre-cooled PBS; then, pre-cooled PBS was used to rinse twice, and pre-cooled trypsin without EDTA was added to digest the cells, followed by addition of digestion stop solution, centrifugation at 1000 rpm for 3 min, washing with pre-cooled PBS, combining the cells collected in the supernatant, discarding PBS after centrifugation, adding Annexin V-FITC and PI staining solutions, staining at room temperature for 15 min, and then immediately using flow cytometry to detect the cell apoptosis status.

[0062] 3. Cell migration assay

[0063] The migration of cells in each group was recorded using an inverted microscope.

[0064] 4. Plate clone detection

[0065] The culture medium was discarded, and the cells were rinsed twice with PBS, then fixed with 4% paraformaldehyde for 15 min and discarded, and rinsed again with PBS twice; crystal violet staining was performed for 15 min, and photographs were taken for record.

[0066] 5. Western Blot Detection

[0067] The cell culture supernatant was discarded, washed twice with PBS, and RIPA (containing 1% PMSF) cell lysis buffer was added to lyse the cells on ice for 30 minutes, collect the cell debris, centrifuge at 12000rpm for 30 minutes, collect the supernatant, quantify the BCA protein, add Loading Buffer, and boil for 10 minutes to prepare the protein sample. Then, the protein immunoblotting experiment was performed, blocked with 5% skim milk for 2 hours, incubated with the corresponding primary antibody at 4°C overnight; washed with 1×TBST for 20 minutes, then incubated with the secondary antibody at room temperature for 2 hours, then washed with 1×TBST for 20 minutes, and developed according to the operating instructions of the developer.

[0068] Results Analysis

[0069] 1. Improve the sensitivity of chemotherapy drugs

[0070] from Figure 1 It can be seen that the sensitization effect of tetrahydropalmatine on paclitaxel under low concentration (5 μM) conditions is manifested as enhancing the inhibitory effect of low concentration paclitaxel and synergistic effect with high concentration paclitaxel. When the concentration of paclitaxel is low (such as 0.00975 μM and 0.0390625 μM), the inhibitory effect of paclitaxel alone on cell viability is limited, and after adding tetrahydropalmatine, cell viability is significantly reduced. When the concentration of paclitaxel is high (such as 10 μM), the addition of tetrahydropalmatine further reduces the cell survival rate to 30.38%, while the effect of paclitaxel alone is not enough to reach this level. This synergistic effect shows that tetrahydropalmatine not only acts on the drug resistance mechanism of tumor cells, but also may improve its killing efficiency by affecting the toxic action target of paclitaxel.

[0071] like Figure 2 As shown, in Example 2, the concentration of tetrahydropalmatine was increased to 50 μM. Although its combination with paclitaxel was still better than paclitaxel alone, the sensitization effect was lower than that of 5 μM. The action of drugs in cells usually manifests as nonlinear effects. The target of tetrahydropalmatine may have reached saturation at lower concentrations. The regulatory effect of paclitaxel concentration further increased under high concentrations of tetrahydropalmatine on the target is no longer significantly enhanced, resulting in the sensitization effect being stabilized or weakened. High concentrations of tetrahydropalmatine may also trigger adaptive responses of cells, such as activating certain protective pathways, thereby partially offsetting its sensitization effect. In addition, although tetrahydropalmatine exhibits a synergistic effect with paclitaxel at low concentrations, a partial antagonistic effect may occur at high concentrations.

[0072] according to Figure 3As a result, in Example 3, under the condition of low paclitaxel concentration (125nM), as the concentration of the combined tetrahydropalmatine increases, the cell viability decreases significantly and shows dose dependence. This reflects the strong correlation between the concentration of tetrahydropalmatine and its sensitizing effect. By using tetrahydropalmatine in combination, the dose of paclitaxel can be significantly reduced, thereby reducing the adverse reactions caused by high-dose chemotherapy and improving the patient's treatment tolerance.

[0073] Figure 4 The data show that the use of tetrahydropalmatine alone in comparative example 1 has a relatively small effect on cell viability. Whether the concentration of tetrahydropalmatine is low (1.5625μM) or high (50μM), the decrease in cell viability is not significant. Tetrahydropalmatine does not show significant concentration-dependent cytotoxicity, indicating that tetrahydropalmatine does not mainly induce cell death directly, but enhances the anti-tumor effect of chemotherapy drugs (such as paclitaxel) through synergistic effects. After combined use with paclitaxel, the viability of drug-resistant cells is significantly reduced, which also shows that the main value of tetrahydropalmatine lies in its sensitization effect, rather than direct toxicity.

[0074] 2. Promote cell apoptosis

[0075] from Figure 5 A It can be seen that in Example 4, the cell morphology of the normal group is complete, polygonal or round, the cell density is high, the adherent growth is good, and no obvious damage or apoptosis characteristics are observed. The cell morphology of the paclitaxel group changes slightly, and some cells appear wrinkled or rounded, indicating that paclitaxel has a certain effect on drug-resistant cells, but the overall impact is relatively limited. Compared with the normal group, the cells in the tetrahydropalmatine group did not change much in morphology, and only a small number of cells appeared slightly rounded or fell off, indicating that when tetrahydropalmatine is used alone, the direct toxic effect on drug-resistant cells is weak. The cell morphology of the tetrahydropalmatine + paclitaxel group is obviously abnormal, manifested as large-area wrinkling, falling off or breaking, and the cell density is significantly reduced, indicating that when tetrahydropalmatine is used in combination with paclitaxel, the killing effect on drug-resistant cells is significantly enhanced, and the combined drug can effectively reverse the drug resistance of paclitaxel and significantly improve the chemotherapy effect.

[0076] according to Figure 5The flow cytometry results of B cell apoptosis showed that most of the cells in the normal group were concentrated in the lower left quadrant, indicating a high proportion of surviving cells, almost no early or late apoptosis, and an extremely low proportion of apoptotic cells. The proportion of surviving cells in the paclitaxel group was basically close to that in the normal group, and the proportion of early and late apoptotic cells was low. The proportion of surviving cells in the tetrahydropalmatine group was slightly reduced, and the proportion of early apoptotic cells was slightly increased, indicating that when tetrahydropalmatine was used alone, the apoptosis-inducing effect on drug-resistant cells was weak, and the direct killing effect was limited. The proportion of surviving cells in the tetrahydropalmatine + paclitaxel group decreased significantly, and the proportion of early apoptotic, late apoptotic and necrotic cells increased significantly. The combined use of drugs significantly increased the proportion of apoptotic cells, indicating that tetrahydropalmatine can significantly enhance the apoptosis-inducing effect of paclitaxel.

[0077] Figure 6 In the results of the plate cloning of each group in Example 6, the number of blue spots in the normal group was the largest, densely distributed, with different sizes of spots, and some spots were larger. The number of blue spots in the tetrahydropalmatine group was similar to that in the normal group, and no significant reduction was observed, indicating that tetrahydropalmatine alone had a weak inhibitory effect on the proliferation of drug-resistant cells and could not significantly reduce the clone formation ability of the cells. The number of blue spots in the paclitaxel group decreased, and the spots were smaller than those in the normal group, but still quite significant, indicating that paclitaxel alone had a limited inhibitory effect on the proliferation of drug-resistant cells. The blue spots in the tetrahydropalmatine + paclitaxel group almost completely disappeared, and no clone formation was observed in the culture dish with the naked eye, indicating that when tetrahydropalmatine was used in combination with paclitaxel, the proliferation ability of drug-resistant cells was significantly inhibited, and the inhibitory effect of the combined drug on clone formation was significantly enhanced.

[0078] Figure 7 In the experiment, PARP (116KD) and its cleavage form Cleaved-PARP (89KD) can reflect the cleavage of PARP protein during apoptosis and are important molecular markers of apoptosis. Bcl-2 (26KD) is an anti-apoptotic protein that inhibits apoptosis, and Bax (21KD) is a pro-apoptotic protein that promotes apoptosis. In Example 5, only the complete PARP band was detected in the normal group, the paclitaxel group, and the tetrahydropalmatine group, and no Cleaved-PARP band was found, indicating that no obvious apoptotic response occurred in the drug-resistant cells. In the tetrahydropalmatine + paclitaxel group, the Cleaved-PARP band was significantly enhanced, while the complete PARP band was weakened. This indicates that the combined use of tetrahydropalmatine and paclitaxel significantly enhanced the apoptotic response of cells, further verifying the sensitizing effect of tetrahydropalmatine on paclitaxel. Compared with the normal group, paclitaxel group and tetrahydropalmatine group, the expression of Bcl-2 in the tetrahydropalmatine + paclitaxel group was significantly decreased and the expression of Bax was significantly upregulated, indicating that the combined use significantly weakened the anti-apoptotic ability of drug-resistant cells while enhancing the pro-apoptotic ability.

[0079] 3. Inhibit cell migration

[0080] from Figure 8 It can be seen that the drug-resistant cells in the normal group, paclitaxel group and tetrahydropalmatine group of Example 5 all migrated from one side of the scratch to the blank area, and the normal group had the strongest cell migration ability, followed by the paclitaxel group. When tetrahydropalmatine was used alone, the healing degree of the scratch area was slightly better than that of the paclitaxel group, indicating that tetrahydropalmatine had a weak inhibitory effect on the migration ability of drug-resistant cells. The scratch area of ​​the tetrahydropalmatine + paclitaxel group was almost not healed, and the cells on both sides of the scratch did not obviously migrate to the blank area, and the blank area remained intact, indicating that the combination of tetrahydropalmatine and paclitaxel had a significant inhibitory effect on the migration ability of drug-resistant cells.

[0081] Obviously, the above embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. For those skilled in the art, other different forms of changes or modifications can be made based on the above description. It is not necessary and impossible to list all the embodiments here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. An application of tetrahydropalmatine in the preparation of chemotherapy sensitization drugs, characterized in that: Used for preparing chemotherapy sensitization drugs; the structural formula of tetrahydropalmatine is:

2. The use of tetrahydropalmatine according to claim 1, characterized in that: The chemotherapy sensitizing drug is used to inhibit cell activity, inhibit cell proliferation, promote cell apoptosis, and inhibit cell migration.

3. The use of tetrahydropalmatine according to claim 2, characterized in that: The cells include cells that are resistant to chemotherapeutic drugs.

4. The use of tetrahydropalmatine according to claim 1, characterized in that: The chemotherapy sensitizer is used to increase the sensitivity of malignant tumors to chemotherapy drugs.

5. The use of tetrahydropalmatine according to claim 4, characterized in that: The malignant tumors include malignant tumors of the female reproductive system.

6. The use of tetrahydropalmatine according to claim 4, characterized in that: The chemotherapy drugs include paclitaxel.

7. A chemotherapy sensitizing drug, characterized in that: The composition includes at least one of tetrahydropalmatine, a tetrahydropalmatine derivative and a pharmaceutically acceptable salt thereof; the structural formula of the tetrahydropalmatine is:

8. The chemotherapy sensitizing drug according to claim 7, characterized in that: The chemotherapy sensitizing drug also includes a chemotherapy drug component.

9. The chemotherapy sensitizing drug according to claim 8, characterized in that: The chemotherapy drug component includes paclitaxel.

10. The chemotherapy sensitizing drug according to claim 9, characterized in that: The molar concentration ratio of tetrahydropalmatine to paclitaxel is 0.5:1 to 1600:1.

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