Traditional Chinese medicine composition for treating glazed pulmonary nodules

Abstract

The traditional Chinese medicine is prepared from 15-20 parts of platycodon grandiflorum, 15-20 parts of selfheal, 10-15 parts of pericarpium citri reticulatae, 10-20 parts of sculellaria barbata, 10-20 parts of oldenlandia diffusa, 12-15 parts of bamboo shavings and 10-15 parts of exocarpium citri grandis. The composition has huge advantages when being used for treating the pure ground glass pulmonary nodules, fills up the window period of regular follow-up visit of medium and low-risk pulmonary nodules without drug treatment, and has the advantages of early intervention, slowing down the progress of nodule malignant transformation, avoiding side effects and medical expenses caused by operations and the like.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and in particular to a traditional Chinese medicine composition for treating ground-glass pulmonary nodules. Background Art

[0002] Lung nodules have now become a major problem threatening the health of Chinese citizens. The Affiliated Hospital of Fudan University has conducted health screening on 11,332 residents in Shanghai. The results showed that more than 70% of residents had ground-glass nodules in their lungs. The detection rate of lung nodules in adults over 18 years old in my country is as high as 55.3%, and the detection rate of ground-glass nodules in the lungs is 7.3%. The risk of malignant transformation of simple ground-glass nodules in the lungs is about 18%, and the probability of mixed ground-glass nodules being diagnosed as malignant is 64%. Ground-glass nodules in the lungs have become a highly concerned imaging pathological manifestation in clinical practice due to their high correlation with lung cancer.

[0003] Pure ground-glass pulmonary nodules refer to ground-glass lesions in the lungs that are less than 30 mm in diameter and do not contain solid components, as screened by chest CT technology. At the tissue cell level, they are characterized by increased cell number, thickened alveolar septa, decreased gas content, but incomplete collapse of the alveoli. Domestic and foreign guidelines and expert consensus recommend that pure ground-glass pulmonary nodules be divided into two risk levels: low risk, medium risk, and high risk. In the case that pure ground-glass pulmonary nodules do not progress, Western medicine currently recommends regular CT follow-up, and no surgery or Western medicine intervention. Traditional Chinese medicine believes that its pathogenesis is closely related to pathogenic factors such as phlegm, blood stasis, and toxins, and the treatment is mainly based on expectoration, blood stasis, and toxin attack. Therefore, the development of a Chinese medicine composition for the treatment of ground-glass pulmonary nodules has become a long-awaited goal. Summary of the invention

[0004] The purpose of the present invention is to provide a traditional Chinese medicine composition for treating ground-glass pulmonary nodules, thereby meeting the above needs.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] In a first aspect, the present invention provides a traditional Chinese medicine composition for treating ground glass pulmonary nodules, the composition comprising the following ingredients in parts by mass: 15-20 parts of Platycodon grandiflorum, 15-20 parts of Prunella Vulgaris, 10-15 parts of Citrus reticulata, 10-20 parts of Scutellaria barbata, 10-20 parts of Hedyotis diffusa, 12-15 parts of Rhizoma Anemarrhenae, and 10-15 parts of Citrus aurantium.

[0007] In a second aspect, the present invention provides use of the above Chinese medicine composition for the manufacture of a medicament for treating ground-glass pulmonary nodules.

[0008] In a third aspect, the present invention provides a medicine for treating ground-glass pulmonary nodules, the medicine comprising the traditional Chinese medicine composition as described above.

[0009] Compared with the prior art, the beneficial effects of the present invention are: the Chinese medicine composition of the present invention has great advantages in treating pure ground glass lung nodules, filling the window period of regular follow-up of medium and low-risk lung nodules without drug treatment, and has the advantages of early intervention, slowing down the progression of nodule malignancy, avoiding side effects and medical expenses caused by surgery, etc. BRIEF DESCRIPTION OF THE DRAWINGS

[0010] Figure 1 The chest CT scan results before and after the use of the composition of the present invention to treat ground glass lung nodules are shown;

[0011] Figure 2 Demonstrate the inhibitory effect of the composition of the present invention on human lung cancer cell lines PC9 and H1975;

[0012] Figure 3 The effects of the composition of the present invention on the PTGS2, MEK1 / 2 and STAT3 signaling pathways in human lung cancer cell lines PC9 and H1975 were demonstrated. DETAILED DESCRIPTION

[0013] The following will be combined with the specific embodiments of the present invention and the accompanying drawings to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0014] In the description of the present invention, it should be noted that the terms "upper", "lower", "inner", "outer", "front end", "rear end", "two ends", "one end", "the other end" and the like indicating the orientation or positional relationship are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance.

[0015] In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "provided with", "connected", etc. should be understood in a broad sense. For example, "connected" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0016] The composition of the present invention comprises the following components by mass: 15-20 parts of Platycodon grandiflorum, 15-20 parts of Prunella vulgaris, 10-15 parts of dried orange peel, 10-20 parts of Scutellaria barbata, 10-20 parts of Hedyotis diffusa, 12-15 parts of Rhizoma Anemarrhenae, and 10-15 parts of Citrus grandiflora. Or each dose of the Chinese medicine composition of the present invention is composed of 15-20 parts of Platycodon grandiflorum, 15-20 parts of Prunella vulgaris, 10-15 parts of dried orange peel, 10-20 parts of Scutellaria barbata, 10-20 parts of Hedyotis diffusa, 12-15 parts of Rhizoma Anemarrhenae, and 10-15 parts of Citrus grandiflora. The composition of the present invention is formulated based on the clinical efficacy on the basis of the ancestral lung treatment theory of the Wang family, and is an effective prescription for treating pure ground glass pulmonary nodules. The prescription is obtained by Wang Peihua, father and son Wang Ning, who are descendants of Wang's traditional Chinese medicine, who have long diagnosed and treated pure ground glass pulmonary nodules and achieved good efficacy, and have accumulated rich experience and have been carefully refined. The main herb of this prescription, Platycodon grandiflorum, is bitter in taste, and its pungent and warm nature can lead all the medicines into the lungs, remove phlegm and pus, and promote the flow of lung qi; Tangerine peel is pungent and warm in taste, and is an important medicine for regulating qi, drying dampness and resolving phlegm. It is combined with Platycodon grandiflorum to enhance the ability of resolving phlegm and dispersing nodules; Herba Scutellariae and Herba Hedyotis diffusae are bitter, pungent, and cold in nature, which can dissipate nodules, reduce swelling and resist inflammation, and at the same time clear liver and gallbladder fire stagnation, regulate qi and dissipate stagnation; Scutellaria barbata and Herba Hedyotis diffusae clear heat and detoxify, and work together with Herba Scutellariae to clear heat and toxins in the lungs, remove blood stasis and dissipate nodules; Rhizoma Anemarrhenae clears heat and resolves phlegm, relieves restlessness and stops vomiting, and the combination of Rhizoma Anemarrhenae and Rhizoma Anemarrhenae can help it clear heat and dry dampness, resolve phlegm and relieve cough; Tangerine peel and Radix Citrus Reticulatae regulate qi and strengthen the spleen, dry dampness and resolve phlegm, and at the same time restrain cold and cool medicines, prevent damage to the spleen and stomach, and can resolve phlegm and dampness to help dissipate nodules. This prescription is well-matched and the medication is sophisticated. It has been used by several generations and has a good effect. This medicine is easy to use, has a reasonable compatibility, and is safe and reliable. It has been widely praised in recent decades.

[0017] The following provides cases and related examples of using the composition of the present invention to treat ground glass lung nodules to further illustrate the present invention.

[0018] Example 1: Clinical observation on 38 cases of pure ground glass pulmonary nodules treated with the composition of the present invention

[0019] The inventor has long been treating pure ground glass pulmonary nodules and achieved good results, accumulating rich experience. During the follow-up period, the patients were treated with Chinese medicine for at least 3 months, and the pure ground glass nodules of some patients were reduced or completely disappeared. This study retrospectively summarizes 38 patients with pure ground glass pulmonary nodules among 360 cases of pulmonary nodules treated by the inventor from June 2022 to September 2024, and summarizes the clinical efficacy and safety of the composition of the present invention in treating pure ground glass pulmonary nodules.

[0020] 1.1 Materials and Methods

[0021] General information: The cases were from 38 patients with pure ground glass pulmonary nodules who visited a hospital outpatient clinic from June 2022 to September 2024. There were 13 males (34.2%) and 25 females (65.8%), with an average age of (54.5±10) years.

[0022] 1.2 Diagnostic criteria

[0023] 1.2.1. Western medicine diagnostic standards: The diagnostic standards of the present invention refer to the definition, diagnosis and classification of lung nodules in the "Chinese Expert Consensus on the Diagnosis and Treatment of Pulmonary Nodules (2018 Edition)" and "Guidelines for the Classification, Diagnosis and Treatment of Pulmonary Nodules in China (2016 Edition)". Classification is based on the following three dimensions: ① Quantity: A single lesion is isolated, and 2 or more lesions are defined as multiple. ② Density: Pure ground glass nodules are confirmed based on density and degree of mixing. ③ Danger level: Pure ground glass nodules are limited by a diameter of 5 mm, with a diameter ≥10 mm (high risk), 5 mm ≤ diameter <10 mm (medium risk), and a diameter <5 mm (low risk).

[0024] 1.2.2. TCM diagnostic standards: There are no TCM disease differentiation and syndrome differentiation standards for pulmonary nodules at present. Patients may have no obvious symptoms or signs, and no syndrome can be diagnosed. In this study, all patients with pulmonary nodules on lung imaging, excluding pulmonary tuberculosis and wind-heat lung heat disease, can be included in the category of TCM "pulmonary nodules".

[0025] 1.3. Inclusion criteria: ① Patients with pulmonary nodules who meet the above diagnostic criteria; ② Patients who only need CT follow-up according to the Chinese guidelines and expert consensus, or patients who need surgery but refuse or are unable to undergo surgery; ③ Age 20 to 90 years old; ④ Continuous treatment with the Chinese medicine composition of the present invention for 3 months.

[0026] 1.4. Exclusion criteria: ① Patients with severe primary diseases such as cardiovascular, liver, kidney, and blood system diseases; ② Patients with active tuberculosis and other infectious diseases; ③ Patients with mental illnesses such as mania, schizophrenia, etc. or unable to cooperate with the observation and research for other reasons; ④ Pregnant or lactating women; ⑤ Patients with missing contact information or incomplete medical records.

[0027] 1.5. Treatment method: Oral treatment with Chinese medicine decoction is given in the outpatient department. The medicine is Chinese medicine slices or Chinese medicine formula granules from the pharmacy of the hospital where the inventor works. One dose per day, each dose of the prescription is basically composed of 15g of Platycodon grandiflorum, 15g of Prunella vulgaris, 12g of dried orange peel, 20g of Scutellaria barbata, 15g of Hedyotis diffusa, 15g of Rhizoma Anemarrhenae, and 15g of Citrus grandiflora. The range of addition and subtraction treatment is 15-20g of Platycodon grandiflorum, 15-20g of Prunella vulgaris, 10-15g of dried orange peel, 10-20g of Scutellaria barbata, 10-20g of Hedyotis diffusa, 12-15g of Rhizoma Anemarrhenae, and 10-15g of Citrus grandiflora. Chinese medicine addition and subtraction treatment for 3 months, during which a follow-up visit is made every 2 to 4 weeks. After 3 months of treatment, a chest CT is reviewed and compared with the results of the most recent chest CT before the initial visit to evaluate the efficacy.

[0028] 1.6. Efficacy criteria: This study used the size of chest CT nodules as the main basis for efficacy, and combined with nodule density, number, etc., to compare before and after treatment. (1) Lung nodule diameter: The maximum diameter of pure ground glass lung nodules shown in the CT imaging report, or the average of the long and short diameters of the nodules in accordance with the Fleischner Association's guidelines for lung nodule measurement. The diameter is expressed by the average of the long and short diameters of the nodules. The reported results and calculated averages are rounded to one decimal place in millimeters (mm). (2) Nodule area of ​​the largest diameter plane of lung nodules in chest CT: The area of ​​the largest diameter plane of the nodule is calculated as S = π × A × B / 4 (ellipse area formula, A and B are the long and short diameters reported in the chest CT report, respectively. If the chest CT report only has the diameter D, then A = B = D, which is the circular area formula). Compare the area reduction rate before and after treatment, and compare the number and density of nodules based on the chest CT report and the film reading. The efficacy is divided into the following levels: ① cured: chest CT shows that the nodules disappear; ② markedly effective: chest CT shows that the nodules become smaller, lighter, and fewer, and the area (area sum) of the largest diameter surface decreases by ≥50%; ③ effective: chest CT shows that the nodules become smaller, lighter, and fewer, and the area (area sum) of the largest diameter surface decreases by <50%; ④ ineffective: chest CT shows that the size, density, and number of nodules are stable without obvious changes or progression. Total effective rate (%) = (number of cured cases + number of markedly effective cases + number of effective cases) / total number of cases × 100%.

[0029] 1.7. Safety evaluation indicators: blood routine, urine routine, stool routine and liver and kidney function before and 3 months after administration, as well as adverse events during treatment were recorded.

[0030] 1.8 Statistical methods: GraphPad prism 9.0 software was used to compare the two sample groups in this study using a two-sided t-test. P < 0.05 was considered statistically significant.

[0031] 1.9 Research Results

[0032] 1) The classification of pure ground glass pulmonary nodules is shown in Table 1.

[0033] For 38 cases of pure ground glass lung nodules, the cutoff was 5 mm, diameter ≥ 10 mm (high risk), 5 mm ≤ diameter < 10 mm (intermediate risk), and < 5 mm (low risk).

[0034] Table 1. Classification of 38 pure ground glass pulmonary nodules (cases)

[0035]

[0036] 2) The efficacy of the composition of the present invention on pure ground glass lung nodules is shown in Table 2.

[0037] The effective rates of treatment for low-, medium-, and high-risk pure ground glass nodules were 25%, 81%, and 73%, respectively, and the total effective rate was 68%. According to statistical tests, the average size of nodules decreased from (70.24±101.9) mm2 to (35.05±46.43) mm2 before and after treatment, with a significant difference (P=0.010). The average size of medium-risk nodules decreased from (27.45±10.66) mm2 to (12.38±13.83) mm2 before and after treatment, with a statistically significant difference (P=0.001). The average size of high-risk nodules decreased from (146.8±143.4) mm2 to (68.17±59.75) mm2 before and after treatment, with a statistically significant difference (P=0.033).

[0038] Table 2. Treatment efficacy of 38 cases of pure ground glass pulmonary nodules

[0039]

[0040] Note: *P<0.05

[0041] 3) Comparison of chest CT scans of patients with pure ground glass nodules before and after treatment with the composition of the present invention

[0042] Figure 1 A representative example is shown by Figure 1 It can be seen that after oral administration of the composition of the present invention, the size of the nodules in the CT images was significantly reduced, from 5.5*3.5 mm before treatment to almost disappearance of the nodules after treatment, and the therapeutic effect was significant.

[0043] 1.10. Safety evaluation indexes: After safety analysis, all patients treated with the composition of the present invention had no adverse events during the treatment period, and no abnormalities were found in the blood routine, liver and kidney function, and urine routine test results of the patients before and after treatment.

[0044] Example 2: Inhibitory effect of the composition of the present invention on lung cancer cells

[0045] The MTT method was used to detect the inhibitory effect of the composition of the present invention on lung cancer cells.

[0046] The specific experimental method is:

[0047] (1) Take the human lung cancer cell lines PC9 and H1975 in the logarithmic growth phase, discard their culture medium, wash twice with 2 mL of PBS, aspirate the PBS, add 1 mL of 0.25% trypsin-0.02% EDTA to infiltrate the cells and then aspirate them, place them in a cell culture incubator for digestion for 1 min, take them out, add 3 mL of complete culture medium to terminate the digestion, and gently blow down the cells to resuspend them;

[0048] (2) After the cells were counted, they were plated in a 96-well cell culture plate, 5,000 cells were added to each well, and cultured overnight in a carbon dioxide cell culture incubator;

[0049] (3) Take the medicinal material slices of the composition of the present invention that meet the requirements of the 2015 edition of the "Chinese Pharmacopoeia", add 8 times the volume of water according to the prescription amount, heat and extract twice, each time for 45 minutes, filter, combine the filtrate, transfer all the liquid medicine to a rotary evaporator by suction, and concentrate it until the volume of the liquid medicine is 100-150ml; after pre-cooling at -80℃ overnight, freeze-dry at -80℃ for 72 hours, grind into powder, package in 50mL centrifuge tubes, and store in a -80℃ ultra-low temperature refrigerator for use. 1g of freeze-dried powder obtained by the above process is equivalent to 9.18g of crude drug. A series of freeze-dried powder solutions of the composition of the present invention with different concentrations were incubated with cells for 24h and 48h, respectively, and a group of control wells without drug addition were set;

[0050] (4) After the culture was completed, 10 μL of 5 mg / mL MTT was added to each well of the 96-well plate and incubated in a CO2 cell culture incubator for 4 h;

[0051] (5) Discard the cell supernatant, add 100 μL of DMSO to each well, shake in the dark for 10 min, measure the absorbance (A) at 570 nm / 630 nm using an ELISA reader and calculate the cell viability;

[0052] (6) Cell viability calculation formula: Cell viability (%) = A value of drug-treated wells / (A value of solvent control wells-A value of blank control wells) × 100%.

[0053] Experimental results:

[0054] In this experiment, human lung cancer cell lines PC9 and H1975 were selected, and the MTT method was used to detect the effects of different concentrations of the composition of the present invention (1.25 mg / mL, 2.5 mg / mL, 5 mg / mL, 10 mg / mL) on the activity of PC9 and H1975 cells. The administration time was 24h and 48h, respectively. Figure 2 The results show that, compared with the control group, the composition of the present invention can significantly inhibit cell proliferation within the concentration range of 2.5-10 mg / mL (*P<0.05, **P<0.01, ***P<0.001).

[0055] Example 3: Effects of the drug-containing serum of the composition of the present invention on PTGS2, MEK1 / 2 and STAT3 signaling pathways

[0056] The process of extracting the drug-containing serum of the composition of the present invention is as follows:

[0057] The extract of the composition of the present invention: add 8 times the volume of water to the medicinal materials of the composition according to the prescribed amount, heat and extract twice, each time for 45 minutes, filter, combine the filtrate, transfer all the medicinal liquid to a rotary evaporator by suction, and concentrate it for later use.

[0058] Animals: SD rats, male. The extract was continuously gavaged for 7 days, the low dose of the rat gavage was 13.9g crude drug / kg, and the daily dose was 10 times the low dose, 2ml / rat, and gavage was performed again one hour before blood collection. Blood was collected from the abdominal aorta of the rats, and after the blood was collected, it was left to stand at 4℃ for 1-2 hours, centrifuged at 3500rpm for 15min, and the supernatant was frozen at -80℃.

[0059] Preparation of drug-containing serum: Take out the serum from -80℃ and inactivate it in a 56℃ water bath for 30 minutes. Filter the inactivated serum through a 0.22μm filter membrane in a clean bench to obtain the drug-containing serum.

[0060] Western blot method

[0061] PC-9 and H1975 cells were cultured at 5 × 10 5 The cells were inoculated in 6-well plates at a density of 100 cells / well. After 24 hours, the drug-containing serum control group (CTL), 10%, 20%, and 30% were added to treat the cells for 24 hours. The cells were harvested to extract proteins, and the protein concentration was detected by BCA method. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed, and the membrane was transferred and blocked. The following antibodies were added: PTGS2 (1:1000), pMEK1 / 2 (1:1000), MEK1 / 2 (1:1000), pSTAT3 (1:1000), STAT3 (1:1000 Cell Signaling Technology, USA) and GAPDH (Bioworld, USA, 1:1000), incubated at room temperature for 2 hours, and then the secondary antibody (1:5000) was added for incubation for 1 hour. Finally, the protein bands were detected using the chemiluminescence (ECL) system (Tianneng, China).

[0062] result

[0063] like Figure 3 As shown, compared with the drug-containing serum control group (CTL), the drug-containing serum (10%, 20%, 30%) of the composition of the present invention can effectively reduce the expression of PTGS2 protein and inhibit the phosphorylation of MEK1 / 2 and STAT3 in PC9 cells, indicating that the composition of the present invention can effectively inhibit the activation of MEK1 / 2 and STAT3 signaling pathways. In H1975 cells, the drug-containing serum (10%, 20%, 30%) of the composition of the present invention can still effectively inhibit the expression of PTGS2 protein and have a certain inhibitory effect on the MEK1 / 2 signaling pathway.

[0064] The matters not described in detail in the present invention are all known technologies to those skilled in the art.

[0065] Finally, it should be noted that the above specific implementation methods are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified and replaced by equivalents without departing from the spirit and scope of the technical solution of the present invention, which should be included in the scope of the claims of the present invention.

Claims

1. A Chinese medicine composition for treating ground glass pulmonary nodules, characterized in that: The composition comprises the following components by mass: 15-20 parts of platycodon, 15-20 parts of selfheal, 10-15 parts of tangerine peel, 10-20 parts of scutellaria barbata, 10-20 parts of oldenlandia diffusa, 12-15 parts of bamboo shavings, and 10-15 parts of tangerine peel.

2. Use of a Chinese medicine composition for the manufacture of a medicament for treating ground-glass pulmonary nodules, characterized in that: The composition comprises the following components by mass: 15-20 parts of platycodon, 15-20 parts of selfheal, 10-15 parts of tangerine peel, 10-20 parts of scutellaria barbata, 10-20 parts of oldenlandia diffusa, 12-15 parts of bamboo shavings, and 10-15 parts of tangerine peel.

3. A drug for treating ground glass pulmonary nodules, characterized in that: The medicine comprises the Chinese medicine composition as claimed in claim 1.

Images