Pharmaceutical composition for treating pulmonary nodule and application thereof

Abstract

The invention discloses a pharmaceutical composition for treating pulmonary nodules, the pharmaceutical composition is composed of a dandelion extract and a ganoderma lucidum extract, and the mass ratio of the dandelion extract to the ganoderma lucidum extract is 1: 0.0625-1: 4. Through experimental research, the inventor finds that the dandelion and the lucid ganoderma are cooperatively used in a specific proportion, so that a remarkable combined synergistic effect on pulmonary nodules is achieved, and the combined synergistic effect is remarkably superior to that of single use of the dandelion or the lucid ganoderma. The medicine disclosed by the invention is simple and convenient to prepare, and can be obtained by jointly decocting the dandelion and the lucid ganoderma, or mixing the dandelion or lucid ganoderma extract and brewing with hot water. The medicine has the advantages of low cost and small side effect.

Description

Technical Field

[0001] The present invention relates to medicine and its application, and in particular to a medicine composition for treating pulmonary nodules and its application. Background Art

[0002] Pulmonary nodules refer to focal round dense shadows of various sizes, with clear or blurred edges and a diameter of less than or equal to 3 cm on lung images. According to the International Early Lung Cancer Action Program, the detection rate of pulmonary nodules in people over 40 years old without clinical symptoms is as high as 30.2%. Another large sample study on pulmonary nodules reported that among the 31% detection rate of pulmonary nodules, about 4% were malignant nodules.

[0003] Persistent lung nodules have a certain probability of being malignant. The probability of malignancy for lung nodules with a diameter of 6 to 8 mm is approximately 7.39%, and the probability of malignancy for lung nodules with a diameter > 8 mm is approximately 21.79%.

[0004] The pain points of treating lung nodules are that it is difficult to distinguish between benign and malignant nodules, the follow-up period is long, and they are easy to evolve into lung cancer, as well as the limitations of existing treatment methods. Western medicine lacks corresponding treatments for the early intervention of lung nodules and the treatment after their malignant transformation. In the early stages, it is mainly based on follow-up management, and there are no targeted effective drugs. Therefore, how to effectively intervene in the progression of lung nodules at an early stage has become a major problem in clinical work. Therefore, it is particularly important to take simple and effective intervention measures to prevent lung nodules in daily life.

[0005] Soaking dried dandelion or ganoderma lucidum materials in hot water will result in low concentration of active ingredients and fail to achieve therapeutic effects. Summary of the invention

[0006] Purpose of the invention: The present invention provides a drug capable of treating lung nodules, which solves the problems of low content of active ingredients, poor efficacy, complicated preparation methods and unstable content of active ingredients in the existing drugs. The pharmaceutical composition is composed of dandelion extract and ganoderma lucidum extract, and the mass ratio of dandelion extract to ganoderma lucidum extract is 16:1 to 1:4 (1:0.0625-1:4). The inventors found that the combination of dandelion extract and ganoderma lucidum extract within a specific ratio range has a significant therapeutic effect on the treatment of lung nodules, which is not only significantly better than the effect of dandelion extract or ganoderma lucidum extract used alone, but also better than the effect of combined use of dried dandelion and ganoderma lucidum medicinal materials.

[0007] Technical solution: A pharmaceutical composition for treating pulmonary nodules, the pharmaceutical composition is composed of dandelion extract and ganoderma lucidum extract, the mass ratio of dandelion extract to ganoderma lucidum extract is 1:0.0625-1:4. Preferably, the mass ratio of dandelion extract to ganoderma lucidum extract is 16:1-15:1.

[0008] In the pharmaceutical composition for treating pulmonary nodules, the mass ratio of the dandelion extract to the ganoderma lucidum extract is 1:0.0625-1:4, the concentration of the dandelion extract is 2-20 mg / mL, and the concentration of the ganoderma lucidum extract is between 1-8 mg / mL.

[0009] In the pharmaceutical composition for treating pulmonary nodules, the mass ratio of the dandelion extract to the ganoderma lucidum extract is 1:0.0625-1:4, the concentration of the dandelion extract is 2-16 mg / mL, and the concentration of the ganoderma lucidum extract is between 1-6 mg / mL. Preferably, the mass ratio of the dandelion extract to the ganoderma lucidum extract is 1:0.0625-1:4, the concentration of the dandelion extract is 2-16 mg / mL, and the concentration of the ganoderma lucidum extract is between 2-6 mg / mL.

[0010] The pharmaceutical composition for treating lung nodules, wherein the dandelion extract is prepared by the following method: soaking dandelion medicinal materials at 70-80°C for 3-4 hours, boiling at 90-100°C for 1-2 hours, and repeatedly extracting 1-5 times, filtering and mixing the extracts, standing for 8-12 hours, discarding the lower layer of sediment and taking the supernatant, and concentrating the solution under reduced pressure rotary evaporation, adding 5-10% maltodextrin to the concentrate, preparing under spray drying, and crushing to obtain the dandelion extract.

[0011] The pharmaceutical composition for treating pulmonary nodules, wherein the ganoderma lucidum extract is prepared by the following method: soaking ganoderma lucidum medicinal materials at 70-80°C for 3-4 hours, boiling at 80-100°C for 1-2 hours, and repeatedly extracting 1-5 times, filtering and mixing the extracts, standing for 8-12 hours, discarding the lower layer of sediment and taking the supernatant, concentrating the solution under reduced pressure rotary evaporation, adding 5-10% maltodextrin to the concentrate, preparing under spray drying, and crushing to obtain ganoderma lucidum water extract.

[0012] The application of the pharmaceutical composition in preparing medicine for treating pulmonary nodules.

[0013] The use of ganoderic acid A and chlorogenic acid in combination in the preparation of a drug for treating pulmonary nodules. Preferably, the molar concentration ratio of chlorogenic acid to ganoderic acid A is 5:1; more preferably, the concentration of chlorogenic acid is 500 μM and the concentration of ganoderic acid is 100 μM.

[0014] In the application, the amount of dandelion extract in a single composition is 1-4 g, and the amount of ganoderma lucidum extract in a single composition is 0.0625-0.4 g.

[0015] A pharmaceutical preparation for treating pulmonary nodules, comprising the pharmaceutical composition and a pharmaceutically acceptable carrier, with or without the addition of pharmaceutical excipients.

[0016] The pharmaceutical preparation for treating pulmonary nodules is in the form of decoction, granules, tablets, capsules, granules or syrups.

[0017] This study verified through experiments that, under some appropriate proportions, chlorogenic acid and ganoderic acid A, as the effective active ingredients of dandelion and ganoderma lucidum, respectively, can indeed achieve significant combined synergistic effects (Table 3). However, if dried dandelion or ganoderma lucidum is soaked in tea only, the water temperature will cool down in a short time after soaking in boiling water, and it is difficult to extract the effective ingredients, which is difficult to achieve the therapeutic effect of pulmonary nodules. There are also problems such as long waiting time for users and difficulty in controlling the leaching time for operators. In addition, when chlorogenic acid is extracted from dry medicinal materials in hot water, there are differences in the leaching time, resulting in differences in the extraction of effective ingredients. For example, when the leaching time is short and the chlorogenic acid concentration is ≤500μM, it is difficult to achieve a higher level of inhibition rate with 100-500μM ganoderic acid A; if the hot water leaching time is long, it will affect the stability of chlorogenic acid, resulting in differences in the effective rate. In addition, when soaking ganoderma lucidum in tea, unless the amount of ganoderma lucidum accounts for a large proportion, the leaching concentration of ganoderic acid A will not reach 100μM. Therefore, the present invention designs a pharmaceutical composition composed of dandelion extract and ganoderma lucidum extract, which can directly achieve therapeutic effects by means of water-soluble brewing and drinking, and is easy to use; the present invention limits the dosage range of the composition, and its dosage is within a reliable range, and the effect of inhibiting lung nodules is outstanding.

[0018] Beneficial effects: Compared with the prior art, the present invention has the following advantages: (1) It is difficult to achieve the therapeutic effect of lung nodules by soaking dried dandelion or ganoderma. On the contrary, by scientifically matching dandelion extract and ganoderma extract, a composition with a significant inhibitory effect on lung nodules can be obtained, indicating that compared with the use of dried dandelion or ganoderma, the use of dandelion extract and ganoderma extract can achieve an unexpected and synergistic therapeutic effect on lung nodules. (2) The pharmaceutical composition of the present invention has low raw material cost, is easy to use, and has little toxic and side effects, which is of great significance for the clinical treatment of lung nodules. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is the inhibitory level of Lewis cells by the combined action of dandelion extract and ganoderma lucidum extract;

[0020] Figure 2 is the inhibitory level of Lewis cells by the combined action of ganoderic acid A and chlorogenic acid;

[0021] Figure 3 Effects of the prescription on lung nodules in mice. DETAILED DESCRIPTION

[0022] The present invention will be further described below in conjunction with the accompanying drawings and specific implementations. It should be understood that the following implementations are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0023] Example 1

[0024] Preparation of dandelion extract: Dandelion medicinal materials are soaked at 70°C for 3 hours, boiled at 90°C for 1 hour, and extracted repeatedly for 3 times. After filtering, the extracts are mixed 3 times and allowed to stand for 8 hours, the lower sediment is discarded, the supernatant is taken, and the solution is concentrated under reduced pressure rotary evaporation. 5% maltodextrin is added to the concentrate, and then prepared under spray drying and crushed to obtain dandelion extract.

[0025] Preparation of Ganoderma lucidum extract: Ganoderma lucidum medicinal materials are soaked at 70℃ for 3 hours, boiled at 80℃ for 1 hour, and extracted repeatedly for 3 times. After filtering, mix the extract 3 times, let it stand for 8 hours, discard the lower precipitate, take the supernatant, concentrate the solution under reduced pressure rotary evaporation, add 5% maltodextrin to the concentrate, prepare it under spray drying, and grind it to obtain Ganoderma lucidum water extract.

[0026] Example 2

[0027] Preparation of dandelion extract: Dandelion medicinal materials are soaked at 70°C for 4 hours, boiled at 100°C for 1 hour, and extracted repeatedly 4 times. After filtering, the mixed extracts are allowed to stand for 8 hours, and then the lower sediment is discarded and the supernatant is taken. The solution is concentrated under reduced pressure rotary evaporation, 10% maltodextrin is added to the concentrate, and then the concentrate is prepared under spray drying and crushed to obtain the dandelion extract.

[0028] Preparation of Ganoderma lucidum extract: Ganoderma lucidum medicinal materials are soaked at 70℃ for 4 hours, boiled at 100℃ for 2 hours, and extracted repeatedly for 4 times. After filtering, the mixed extracts are allowed to stand for 8 hours, and then the lower precipitate is discarded and the supernatant is taken. The solution is concentrated under reduced pressure rotary evaporation, 10% maltodextrin is added to the concentrate, and then prepared under spray drying, and the Ganoderma lucidum water extract is obtained after pulverization.

[0029] Example 3

[0030] Preparation of dandelion extract: Dandelion medicinal materials are soaked at 80°C for 4 hours, boiled at 100°C for 2 hours, and extracted repeatedly for 3 times. After filtering, the extracts are mixed 3 times and allowed to stand for 12 hours, the lower sediment is discarded, the supernatant is taken, and the solution is concentrated under reduced pressure rotary evaporation. 10% maltodextrin is added to the concentrate, and the concentrate is prepared under spray drying and crushed to obtain the dandelion extract.

[0031] Preparation of Ganoderma lucidum extract: Ganoderma lucidum medicinal materials are soaked at 80℃ for 4h, boiled at 100℃ for 2h, and extracted repeatedly for 3 times. After filtering, mix the extract 3 times, let it stand for 12h, discard the lower precipitate, take the supernatant, concentrate the solution under reduced pressure rotary evaporation, add 10% maltodextrin to the concentrate, prepare it under spray drying, and grind it to obtain Ganoderma lucidum water extract.

[0032] Example 4

[0033] 1 Reagents and Materials

[0034] ① Extract

[0035] The dandelion and ganoderma extracts were provided by Nanjing Qinling Pharmaceutical Technology Co., Ltd. (Examples 1-3). The extracts were prepared with culture medium, filtered with a 0.22 μm sterile filter, and packaged.

[0036] ② Cell lines

[0037] Mouse Lewis cells were provided by Jiangsu Key Laboratory of Tumorigenesis and Intervention.

[0038] ③Cell culture reagents

[0039] DMEM medium (Keygen Biotechnology) and fetal bovine serum (Sijiqing) were mixed at a ratio of 10:1 when used.

[0040] ④Related indicator detection kit

[0041] MTT solution: Protect from light, dissolve 0.05 g MTT (Yisheng Biotechnology, Shanghai, China) powder in 10 mL PBS buffer, sonicate for 20 min, and prepare a 5 mg / mL solution. Filter with a 0.22 μm sterile filter, aliquot, and store at -80 °C. Dissolve at room temperature before use.

[0042] 2 Experimental methods

[0043] (1) Cell viability detection: The principle is that MTT can be reduced by intracellular mitochondrial dehydrogenase to generate blue-purple crystalline formazan. The optical density (OD) value is detected and compared at a wavelength of 570nm, which can reflect the relative level of cell viability. The operation process is that the cells are evenly cultured in a 96-well plate. When the density reaches 70%, the corresponding concentration of monomer compounds or extracts are added to make the two drugs reach the final effective concentration. The cells are cultured in a 37°C, 5% C02 incubator; after the drug is exposed for 48 hours, 15μL of MTT solution is added to each well. After incubation in the incubator for 3 hours, the OD value at 570nM is measured using an enzyme marker, and the cell viability before and after drug administration can be calculated. Inhibition rate = (1-average absorbance value of cells in the drug group / average absorbance value of cells in the control group).

[0044] (2) Combination index (CI) analysis: The median effect principle was used and the calculation was performed using Compusyn software. The average CI interval and interaction evaluation are shown in Table 1.

[0045] Table 1 Average CI value intervals and interaction evaluation of drug combination effects

[0046]

[0047] 3 Experimental results

[0048] like Figure 1 As shown in the results, when acting alone, dandelion extract significantly inhibited the activity of Lewis cells at a concentration higher than 16 mg / mL; Ganoderma lucidum extract inhibited the activity of Lewis cells in a concentration-dependent manner at 1-8 mg / mL. The combined action of the two drugs can increase the level of inhibition of Lewis cell activity. As shown in Table 2, after calculating the synergistic index, it can be seen that the dandelion extract: Ganoderma lucidum extract mass ratio of 16:1 to 1:4 (1:0.0625-1:4) has a medium or above (bold in the table) combined inhibitory effect on Lewis cell activity. From the results, it can be seen that under the combined action of the two extracts, the concentration range of achieving a medium or above combined synergistic effect is wide, the applicability is strong, the inhibitory effect is outstanding, and the inhibition level reaches at least 51.19%.

[0049] Table 2 Inhibitory level and synergistic effect of dandelion extract and Ganoderma lucidum extract on Lewis cells

[0050]

[0051]

[0052] Example 5

[0053] Chlorogenic acid was provided by Felix; ganoderic acid A was provided by Bede Pharmaceuticals. Before use, compound powder was prepared into a 0.05M stock solution with dimethyl sulfoxide and stored at -80°C. When used, it was diluted to the required concentration with the corresponding cell culture medium.

[0054] Other reagents, materials and experimental methods are the same as those in Example 4.

[0055] 1 Experimental results

[0056] Ganoderic acid A, an abundant component in Ganoderma lucidum, and chlorogenic acid, an abundant component in Taraxacum officinale, were selected to detect the inhibitory level of the two monomer compounds acting simultaneously on Lewis cells. Figure 2As shown in the figure, the two compounds inhibited the activity level of Lewis cells at a low level. However, when the two compounds were used in combination, the level of Lewis cell viability inhibition could be increased, especially when the chlorogenic acid reached 1500μM, the inhibition level was higher. As shown in Table 3, after calculating the synergistic index, it can be seen that a moderate synergistic effect can be obtained when the molar concentration ratio of chlorogenic acid and ganoderic acid A is in the range of 5:1, and the concentrations of chlorogenic acid 500μM and ganoderic acid 100μM. However, the concentrations of ganoderic acid A and chlorogenic acid must reach a more stringent concentration ratio to achieve a synergistic effect; and when chlorogenic acid is extracted with hot water in the form of dry medicinal materials, there is a problem that the immersion time is short and the effective extraction concentration cannot be reached. When the chlorogenic acid concentration is ≤500μM, it is difficult to achieve a higher level of inhibition rate with 100-500μM ganoderic acid A; and a long hot water extraction time affects the stability of chlorogenic acid, and will cause users to wait for a long time, cannot be used in time, and operators are not easy to control the extraction time.

[0057] Table 3 Inhibitory level and synergistic effect of ganoderic acid A and chlorogenic acid on Lewis cells

[0058]

[0059]

[0060] Example 6

[0061] 1 Reagents and Materials

[0062] ①Medication

[0063] Dandelion and Ganoderma lucidum extracts were provided by Nanjing Qinling Pharmaceutical Technology Co., Ltd. Referring to the maximum difference ratio (16:1) in Example 4 for achieving a strong combined synergistic effect, for ease of application, dandelion and Ganoderma lucidum extracts were mixed at a ratio of 15:1 in the in vivo experiment for the verification of lung nodules in vivo. According to the equivalent dose ratio of mice to humans of 9.1 (note: the dose is calculated in mg / kg) and the daily drinking volume of 500 mL for humans, 1.125 g of dandelion and Ganoderma lucidum extracts were mixed, 20 mL of purified water was added, and dissolved (Prescription 1).

[0064] ② Animals

[0065] Source, strain, and line: C57BL / 6 mice were provided by Hangzhou Medical College (production) (experimental animal production license: SCXK (Zhejiang) 2024-0002).

[0066] Age: 35-40 days

[0067] Weight: 18-20g

[0068] Gender: Female

[0069] ③ Pulmonary nodule modeling

[0070] Mouse Lewis lung cancer cell transplants were established by inoculating Lewis lung cancer cell lines into the tail vein of mice. The cell inoculation amount was 1×10 6 Lewis lung cancer cell lines of mice in the logarithmic growth phase were taken and prepared into a cell suspension under sterile conditions, and 0.1 mL was inoculated into the tail vein of the mice.

[0071] ④ Administration: 0.4 mL of Prescription 1 was administered by gavage to mice with pulmonary nodule model. Administration began on the second day after inoculation, and the mice were killed after 18 days of administration, and the lungs were removed for observation.

[0072] 2 Experimental results

[0073] Prescription 1 had a very significant inhibitory effect on mouse lung nodules ( Figure 3 , Table 4), but had no significant effect on the body weight of mice (Table 5).

[0074] Table 4 Effects of prescription 1 on mouse lung nodules (X+SD)

[0075]

[0076] Table 5 Effects of prescription 1 on the body weight of mice

[0077]

Claims

1. A pharmaceutical composition for treating pulmonary nodules, characterized in that: The pharmaceutical composition consists of dandelion extract and ganoderma lucidum extract, and the mass ratio of the dandelion extract to the ganoderma lucidum extract is 1:0.0625-1:

4.

2. The pharmaceutical composition for treating pulmonary nodules according to claim 1, characterized in that: The concentration of the dandelion extract is 2-20 mg / mL, and the concentration of the ganoderma lucidum extract is between 1-8 mg / mL.

3. The pharmaceutical composition for treating pulmonary nodules according to claim 1, characterized in that: The concentration of the dandelion extract is 2-16 mg / mL, and the concentration of the ganoderma lucidum extract is between 1-6 mg / mL.

4. The pharmaceutical composition for treating pulmonary nodules according to claim 1, characterized in that: The dandelion extract is prepared by the following method: soaking dandelion medicinal materials at 70-80°C for 3-4 hours, boiling at 90-100°C for 1-2 hours, and repeatedly extracting 1-5 times, filtering and mixing the extracts, standing for 8-12 hours, discarding the lower layer of sediment, taking the supernatant, and concentrating the solution under reduced pressure rotary evaporation, adding 5-10% maltodextrin to the concentrate, preparing under spray drying, and crushing to obtain the dandelion extract.

5. The pharmaceutical composition for treating pulmonary nodules according to claim 1, characterized in that: The ganoderma lucidum extract is prepared by the following method: soaking ganoderma lucidum medicinal materials at 70-80°C for 3-4 hours, boiling at 80-100°C for 1-2 hours, and repeatedly extracting 1-5 times, filtering and mixing the extracts, standing for 8-12 hours, discarding the lower layer of sediment and taking the supernatant, concentrating the solution under reduced pressure rotary evaporation, adding 5-10% maltodextrin to the concentrate, preparing under spray drying, and crushing to obtain the ganoderma lucidum water extract.

6. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of a drug for treating pulmonary nodules.

7. Application of ganoderic acid A and chlorogenic acid in the preparation of drugs for the treatment of pulmonary nodules.

8. The use according to claim 7, characterized in that: The amount of dandelion extract in a single composition is 1-4 g, and the amount of ganoderma lucidum extract in a single composition is 0.0625-0.4 g.

9. A pharmaceutical preparation for treating pulmonary nodules, characterized in that: The pharmaceutical preparation comprises the pharmaceutical composition according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier, with or without the addition of pharmaceutical excipients.

10. The pharmaceutical preparation for treating pulmonary nodules according to claim 9, characterized in that: The dosage form of the pharmaceutical preparation is decoction, granule, tablet, capsule, granule or syrup.

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