An antitumor composition, a medicine for preventing and / or treating a tumor, and use

Abstract

The application belongs to the technical field of biological medicine, and particularly relates to an anti-tumor composition, a medicine for preventing and / or treating tumors and application. The anti-tumor composition provided by the application comprises dihydroquercetin and beta-carotene; the dihydroquercetin and beta-carotene combined preparation can inhibit the proliferation and growth of various tumor cells of breast cancer, liver cancer, gastric cancer, prostate cancer, lung cancer and colorectal cancer, has an application prospect for preventing and / or treating various cancers, and meanwhile, taking breast cancer and colorectal cancer as examples, it is proved that the dihydroquercetin and beta-carotene can inhibit the tumor growth and metastasis of breast cancer and colorectal cancer. It can be seen that the composition provided by the application can significantly inhibit the proliferation, growth and metastasis of tumors, and can be used for preparing a medicine for preventing and / or treating primary and metastatic tumors.

Description

Technical Field

[0001] This invention belongs to the field of biomedical technology, specifically relating to an anti-tumor composition, a drug for the prevention and / or treatment of tumors, and its application. Background Technology

[0002] Cancer incidence and mortality rates are currently on the rise, with common cancers including lung cancer, colorectal cancer, pancreatic cancer, cervical cancer, and breast cancer. For example, breast cancer is a benign or malignant tumor caused by abnormal proliferation of mammary epithelial cells. Globally, breast cancer ranks second in incidence, accounting for 11.6% of all cancers, with a mortality rate of 6.6%. Despite recent advances in cancer diagnosis, prevention, and treatment, it remains a global health problem affecting millions. Common prevention and treatment methods include chemotherapy, surgery, radiotherapy, and hormone therapy. However, these methods can cause adverse reactions and multidrug resistance, leading to complications. Due to the abundance and diversity of natural products, their multi-target mechanisms of action, and good safety profiles, there is an increasing demand for novel anticancer drugs derived from medicinal plants and natural products. Currently, there are no reports on the combined use of dihydroquercetin and carotene for the prevention and treatment of tumors in existing technologies. Summary of the Invention

[0003] This invention provides an antitumor preparation, a drug for the prevention and / or treatment of tumors, and its application. The antitumor preparation of this invention includes dihydroquercetin and β-carotene, which can significantly inhibit the proliferation of various tumors and can be used to prepare drugs for the prevention and treatment of primary and metastatic tumors.

[0004] To address the above problems, the present invention provides the following technical solution:

[0005] The present invention provides an antitumor composition comprising: dihydroquercetin and β-carotene.

[0006] Preferably, the mass ratio of dihydroquercetin to β-carotene in the composition is (10:1) to (1:10).

[0007] The present invention provides the use of the composition described in the above technical solution in the preparation of drugs for the prevention and / or treatment of tumors.

[0008] Preferably, the application of the drug includes any one or more of the following:

[0009] (1) Inhibits the growth of tumor cells;

[0010] (2) Reduce the frequency of metastatic tumors;

[0011] (3) Reduce the number of distant metastatic lesions of metastatic tumors.

[0012] Preferably, the tumor includes one or more of breast cancer, liver cancer, stomach cancer, prostate cancer, lung cancer, and colorectal cancer.

[0013] The present invention provides a medicament for the prevention and / or treatment of tumors, comprising the composition described above and pharmaceutically acceptable excipients.

[0014] Preferably, the dosage form of the drug includes one or more of oral dosage forms, injections, and suppositories.

[0015] Beneficial Effects of the Invention: This invention provides an antitumor composition comprising dihydroquercetin and β-carotene. The invention utilizes a combined formulation of dihydroquercetin and β-carotene to exhibit significant killing effects on various types of tumor cells, including breast cancer cells, liver cancer cells, gastric cancer cells, prostate cancer cells, lung cancer cells, and colorectal cancer cells. Example results show that monitoring tumor growth and distant organ metastasis in MMTV-PyVT transgenic mice with spontaneous breast cancer demonstrates that the combined formulation of dihydroquercetin and β-carotene can inhibit tumor cell growth, reduce the frequency of metastatic tumors, and decrease the number of distant metastatic lesions. It was also verified that the volume of colorectal cancer tumors in mice in the dihydroquercetin and β-carotene combined formulation group was significantly reduced. Therefore, the composition of this invention can significantly inhibit tumor proliferation, growth, and metastasis, and can be used to prepare drugs for the prevention and treatment of tumors. Furthermore, compared with the use of dihydroquercetin or β-carotene alone, the combined formulation of dihydroquercetin and β-carotene in this invention exhibits a more significant ability to inhibit tumor growth and metastasis.

[0016] The compositions of the present invention, containing dihydroquercetin and β-carotene, are both derived from natural products, exhibiting low toxicity, high activity, few side effects, and a wide range of applications. Attached Figure Description

[0017] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the embodiments will be briefly described below.

[0018] Figure 1 The combined formulation of dihydroquercetin and β-carotene exhibits inhibitory effects on the proliferation of various tumor cells;

[0019] Figure 2 The graph shows the effects of the combined formulation of dihydroquercetin and β-carotene on tumor volume and body weight in spontaneously breast cancer mice.

[0020] Figure 3 The figure shows the effect of the combined preparation of dihydroquercetin and β-carotene on lung metastases of breast cancer.

[0021] Figure 4Figure showing the effects of dihydroquercetin or β-carotene alone on tumor growth and metastasis in spontaneous breast cancer mice;

[0022] Figure 5 This figure shows the changes in tumor growth in a PDX model mouse with colorectal cancer caused by the combined preparation of dihydroquercetin and β-carotene. Detailed Implementation

[0023] The present invention provides an antitumor composition comprising: dihydroquercetin and β-carotene.

[0024] This invention does not have any particular limitation on the source of the dihydroquercetin and β-carotene; conventional commercially available products are sufficient. Preferably, the dihydroquercetin and β-carotene can be extracted from natural substances or synthesized artificially based on their chemical properties.

[0025] The structural formula of dihydroquercetin (DHQ) described in this invention is shown in Formula 1. As a dihydroflavonol compound, dihydroquercetin possesses a wide range of biological activities, including anti-inflammatory, antioxidant, cardiovascular protective, antibacterial, and antiviral effects. Dihydroquercetin has low toxicity to humans and is found in various herbal medicines. In this embodiment of the invention, the dihydroquercetin described in this invention was purchased from Shanghai Biede Pharmaceutical Technology Co., Ltd., product number BD43158.

[0026]

[0027] The β-carotene (C) of this invention 40 H 56 The structural formula of β-carotene is shown in Formula 2. β-carotene is a natural pigment with a wide range of physiological functions, exhibiting excellent anti-cancer effects. It inhibits the growth of tumor cells and induces their apoptosis, thereby increasing the level of reactive oxygen species within tumor cells. In this embodiment of the invention, the β-carotene was purchased from Shanghai Bied Pharmaceutical Technology Co., Ltd., product number BD116471.

[0028]

[0029] In this invention, the mass ratio of dihydroquercetin to β-carotene in the composition is preferably (10:1) to (1:10), more preferably (5:1) to (1:5), even more preferably (2:1) to (1:2), and more preferably 1:1.

[0030] This invention provides the use of the antitumor composition described in the above-mentioned technical solution in the preparation of drugs for the prevention and / or treatment of tumors. The tumors mentioned in this invention include one or more of breast cancer, liver cancer, gastric cancer, prostate cancer, lung cancer, and colorectal cancer. The application of the drug includes any one or more of the following: (1) inhibiting the growth of tumor cells; (2) reducing the frequency of metastatic tumors; (3) reducing the number of distant metastatic lesions.

[0031] The present invention provides a medicament for the prevention and / or treatment of tumors, comprising the antitumor composition described above and pharmaceutically acceptable excipients.

[0032] The dosage form of the drug includes one or more of the following: oral dosage form, injection, and suppository.

[0033] To further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below with reference to the accompanying drawings and embodiments, but these should not be construed as limiting the scope of protection of the present invention.

[0034] Dihydroquercetin (DHQ) was purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd., product number BD43158;

[0035] β-carotene, purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd., product number BD116471.

[0036] Example 1

[0037] The combined preparation of dihydroquercetin and β-carotene inhibits the proliferation of multiple types of tumor cells.

[0038] The total concentration of the composition was 25 μg / mL as the drug concentration. The drug was prepared by dissolving 5 mg of dihydroquercetin and 5 mg of β-carotene in 1 mL of dimethyl sulfoxide to obtain a stock solution with a total concentration of 10 mg / mL. The stock solution was then diluted with cell culture medium to obtain a working concentration of 25 μg / mL for cell treatment. The mass ratio of dihydroquercetin to β-carotene in the drug was 1:1.

[0039] Experimental method: CCK-8 assay was used to detect changes in cell proliferation rate.

[0040] Experiment 1:

[0041] MDA-MB231 breast cancer cells were seeded into 96-well plates at a cell density of 5 × 10⁶ cells per well. 3 Cells. 96-well plates were incubated in a 37°C constant temperature and humidity incubator containing 5% CO2 for 16 hours.

[0042] The treatment group consisted of cells treated with a drug concentration of 25 μg / mL in 96-well plates, with 3 replicates.

[0043] The blank control group consisted of cell culture medium of the same drug volume as the treatment group added to a 96-well plate, with 3 replicates.

[0044] The control group consisted of cells treated with 25 μg / mL dimethyl sulfoxide (DMSO) in 96-well plates, with three replicates. The volume of DMSO added was the same as that added to the treatment group.

[0045] After culturing cells in each group for 48 hours, 10 μL of CCK-8 reagent was added directly to the resulting cell culture medium, and the cells were returned to the cell culture incubator for another 3 hours of incubation. The absorbance (OD) value at a wavelength of 450 nm was measured using a microplate reader. The cell viability of each group was calculated using the formula: Cell viability (%) = (OD value of treatment group - OD value of blank group) / (OD value of control group - OD value of blank group) × 100%.

[0046] Experiment 2: Same as Experiment 1, except that MDA-MB231 breast cancer cells were replaced with HepG2 liver cancer cells;

[0047] Experiment 3: Same as Experiment 1, except that MDA-MB231 breast cancer cells were replaced with HGC-27 gastric cancer cells;

[0048] Experiment 4: Same as Experiment 1, except that MDA-MB231 breast cancer cells were replaced with PC3 prostate cancer cells;

[0049] Experiment 5: Same as Experiment 1, except that MDA-MB231 breast cancer cells were replaced with A549 lung cancer cells;

[0050] Experiment 6: Same as Experiment 1, except that the MDA-MB231 breast cancer cells were replaced with HCT116 colorectal cancer cells.

[0051] The CCK-8 detection results for each experiment are as follows: Figure 1 As shown, the combined preparation of dihydroquercetin and β-carotene can effectively inhibit the growth of various tumor cells, indicating that the combined preparation of dihydroquercetin and β-carotene has broad anti-tumor activity. The specific inhibitory effects on cell viability of different cell types are as follows: MDA-MB231 (control group 100%, treatment group 44.8%), HepG2 (control group 100%, treatment group 70.4%), HGC-27 (control group 100%, treatment group 61.9%), HCT116 (control group 100%, treatment group 40.4%), A549 (control group 100%, treatment group 52.8%), and PC3 (control group 100%, treatment group 60.5%).

[0052] Example 2

[0053] 1. Effects of a combined preparation of dihydroquercetin and β-carotene on breast cancer tumor growth and metastasis.

[0054] Ten 7-week-old female MMTV-PyVT mice were used. The mice were housed in an environment with a relative humidity of 45%–55%, a controlled temperature of 20–25°C, and 12 hours of light / day followed by 12 hours of darkness. The mice were randomly divided into two groups: a control group (con) and a treatment group (treat), with five mice in each group. The control group was administered 0.4 mL of distilled water by gavage, while the treatment group was administered a dihydroquercetin and β-carotene aqueous solution (referred to as di-β-aqueous solution) by gavage. The di-β-aqueous solution was prepared by dissolving 200 mg of dihydroquercetin and 200 mg of β-carotene in 40 mL of sterile distilled water, with a dimethyl sulfoxide concentration of 0.05 mL / mL in the 40 mL of sterile distilled water.

[0055] Mice in each group were administered 0.2 mL of di-β-water solution per 20 g of mouse body weight via gavage once daily for 8 consecutive weeks; tumor size and body weight were measured weekly.

[0056] After 8 weeks of drug administration, all mice were euthanized, and vital organs were collected for analysis. The t-test was used to assess the differences between the two groups. The differences were considered statistically significant when *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0057] Experimental results

[0058] (1) Measurement of tumor volume and body weight changes in mice in each experimental animal group

[0059] The formula for calculating the tumor inhibition rate is: Tumor inhibition rate (%) = (1 - Tumor weight in the treatment group / Tumor weight in the control group) × 100%

[0060] The combined formulation of dihydroquercetin and β-carotene significantly inhibited the growth of breast cancer tumors in MMTV-PyVT transgenic mice, achieving an inhibition rate of 90.8%. Experimental results are shown below. Figure 2 In the figures A, B, and C, C represents the tumor volume of each mouse in the two groups after dissection. It can be seen that after 8 weeks of drug administration, the tumor volume was significantly lower than that of the control group (2142.93 ± 379.54 mm). 3 Compared to the group treated with a combination of dihydroquercetin and β-carotene (tumor volume 196.91±120.81 mm), the group treated with the combined dihydroquercetin and β-carotene (tumor volume 196.91±120.81 mm) 3The tumor volume of mice was significantly reduced (P < 0.0001), while there was no significant difference in body weight (con, 36.64 ± 2.17 g; treat, 26.3 ± 1.26 g).

[0061] (2) HE staining to detect lung metastasis in MMTV PyVT breast cancer model mice

[0062] After euthanizing the mice, the lungs were removed, rinsed thoroughly, and fixed in 4 g / mL paraformaldehyde solution for 24 h. The lungs were then dehydrated, routinely embedded in paraffin, and sectioned. Hematoxylin-eosin staining was performed for 5 min, followed by dehydration and mounting with neutral resin.

[0063] HE staining results of mouse lung tissue are as follows Figure 3 As shown: Mice in the control group had visible metastatic tumors in their lungs. After 8 weeks of combined intervention with dihydroquercetin and β-carotene, the size and number of metastatic tumors were significantly reduced. Compared with the control group (10.33±1.58 tumors), the combined treatment group had only 1.29±0.71 tumors (P<0.01). The number of mice with lung metastases was also significantly reduced (P<0.0001). All mice in the control group developed lung metastases, while only 40% of the mice in the combined treatment group did. This demonstrates that the combined dihydroquercetin and β-carotene treatment effectively inhibits lung metastases in breast cancer.

[0064] 2. Compare the effects of the combined formulation of dihydroquercetin and β-carotene versus dihydroquercetin or β-carotene alone on tumor growth and metastasis in mice.

[0065] (1) A total of 24 female MMTV-PyVT mice, 7 weeks old, were housed in an environment with a relative humidity of 45%–55%, a controllable temperature of 20–25℃, 12 h / d light, and 12 h / d darkness. The mice were randomly divided into 4 groups: control group, combination preparation group, dihydroquercetin group, and β-carotene group, with 6 mice in each group. The control group mice were administered distilled water by gavage, while the combination preparation group mice were administered dihydroquercetin and β-carotene aqueous solution (referred to as di-β-aqueous solution) by gavage. The di-β-aqueous solution was prepared by dissolving 200 mg of dihydroquercetin and 200 mg of β-carotene in 40 mL of sterile distilled water, with a dimethyl sulfoxide concentration of 0.05 mL / mL in the 40 mL of sterile distilled water.

[0066] The dihydroquercetin group was given dihydroquercetin solution by gavage. The dihydroquercetin solution was prepared by dissolving 200 mg of dihydroquercetin in 40 mL of sterile dimethyl sulfoxide distilled water. The volume concentration of dimethyl sulfoxide in the sterile distilled water was 0.05 mL / mL.

[0067] The β-carotene group was administered β-carotene solution by gavage. The β-carotene solution was prepared by dissolving 200 mg of β-carotene in 40 mL of sterile dimethyl sulfoxide distilled water. The volume concentration of dimethyl sulfoxide in the sterile distilled water was 0.05 mL / mL.

[0068] The mice in the above four groups were administered the solution once daily by gavage. The control group was administered 0.4 mL of distilled water by gavage, while the combined preparation group, dihydroquercetin group, and β-carotene group were administered 0.2 mL of solution per 20 g of mouse body weight by gavage for 8 consecutive weeks. The tumor size and body weight of the mice in the four groups were measured weekly.

[0069] After the experiment, all mice were euthanized, and vital organs were collected for analysis and testing. The t-test was used to assess the differences between the two groups. The differences were considered statistically significant when *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001.

[0070] The experimental results are shown in Figure 4 :

[0071] Eight weeks after drug intervention, compared with the control group mice (con), the tumor volume was 2041.01±1039.84 mm. 3 Compared to tumors with a tumor size of 4.21±1.32g (tumor weight), those treated with β-carotene alone had a tumor volume of 1022.80±517.66mm. 3 The tumor weight was 3.98±1.11g, which did not effectively inhibit tumor growth in mice. Although oral administration of dihydroquercetin (DHQ) alone could not effectively inhibit tumor growth, the tumor volume was 495.56±167.21mm. 3 The tumor weight was 1.28±0.46g, and the tumor volume was 119.07±46.99mm. (Tumor weight 1.28±0.46g) and a combination of dihydroquercetin and β-carotene were also tested. 3Both the tumor weight (0.77±0.38g) and the combined preparation of dihydroquercetin and β-carotene significantly inhibited the growth of breast cancer tumors in transgenic mice (P<0.05; P<0.001). However, compared with the dihydroquercetin group, the combined preparation of dihydroquercetin and β-carotene showed a more significant anti-tumor effect (P<0.01). In the prevention and treatment of lung metastases from breast cancer, after 8 weeks of drug intervention, the average number of metastatic lesions in the control group, the combined preparation group, the dihydroquercetin group, and the β-carotene group were 16.16±3.58, 1.28±0.71 (P<0.01), 5.16±3.15 (P<0.05), and 11.16±2.15, respectively. Meanwhile, the proportions of mice developing lung metastases in the control group, the combined preparation group, the dihydroquercetin group, and the β-carotene group were con, 100%; combination, 43.9%; DHQ, 80%; and β-carotene, 100%, respectively. This indicates that the combined preparation of dihydroquercetin and β-carotene has a significant advantage in the prevention and treatment of breast cancer.

[0072] Example 3

[0073] Effects of the combined preparation of dihydroquercetin and β-carotene on the growth of colorectal cancer

[0074] 1. Experimental Methods

[0075] Measurement of tumor volume and body weight changes in mice in different experimental animal groups

[0076] Ten male NOD-SCID mice, 7 weeks old, were used. The mice were housed in an environment with a relative humidity of 45–55% and a controlled temperature of 20–25°C, using a 12-hour / day light / dark cycle. Fresh colorectal tumor tissue was obtained and cut into small pieces. The tumors were inoculated into the ventral region of the mice, and tumor formation was observed daily after inoculation. After tumor formation was observed on the animal's body surface, the tumor-bearing mice were randomly divided into two groups (n=5 per group): a control group and a combination therapy group. Each mouse in the control group was administered 0.4 mL of distilled water by gavage once daily for 8 weeks. Mice in the combination therapy group were administered a solution of dihydroquercetin and β-carotene by gavage. The dihydroquercetin and β-carotene solution was prepared by dissolving 200 mg of dihydroquercetin and 200 mg of β-carotene in 40 mL of sterile distilled water, with a dimethyl sulfoxide concentration of 0.05 mL / mL. This solution was administered by gavage once daily at a volume of 0.2 mL of dihydroquercetin solution per 20 g of mouse body weight for 8 weeks. Tumor size and body weight were measured weekly in both the control and combination therapy groups.

[0077] 2. Experimental Results:

[0078] The combined formulation of dihydroquercetin and β-carotene significantly inhibited tumor growth in a colorectal cancer PDX model mouse, achieving an inhibition rate of 85.3% after 8 weeks of administration. Experimental results are as follows... Figure 5 As shown, tumor volume was measured in all mice 8 weeks after drug intervention. Figure 5 The right figure shows the tumor volume of 5 mice in each of the two groups after dissection, compared with the control group (957.07±253.28 mm). 3 Compared to the dihydroquercetin and β-carotene combination group (139.90±51.60 mm), the combined formulation group... 3 The tumor volume of mice was significantly reduced (P < 0.001), indicating that the combined preparation of dihydroquercetin and β-carotene can inhibit the growth of colorectal cancer.

[0079] In summary, the dihydroquercetin and β-carotene composition provided by this invention inhibits the growth and metastasis of various tumor cells.

[0080] Although the above embodiments have provided a detailed description of the present invention, they are only some embodiments of the present invention, and not all embodiments. People can obtain other embodiments based on these embodiments without creative effort, and these embodiments all fall within the protection scope of the present invention.

Claims

1. An antitumor composition, characterized in that, The composition is made from dihydroquercetin and β-carotene; The mass ratio of dihydroquercetin to β-carotene is 1:

1.

2. The use of the composition according to claim 1 in the preparation of a drug for the prevention and treatment of breast cancer.

3. A drug for the prevention and treatment of breast cancer, characterized in that, The drug is prepared from the composition of claim 1 and pharmaceutically acceptable excipients.

4. The drug according to claim 3, characterized in that, The dosage form of the drug is selected from oral dosage forms, injections, and suppositories.

Images