Pharmaceutical composition and application thereof

Abstract

Disclosed is a pharmaceutical composition comprising (a) a compound of formula (I); the traditional Chinese medicine composition comprises the following raw materials in parts by weight: 50-250 parts of peach kernels, 300-700 parts of salvia miltiorrhiza, 200-600 parts of gynostemma pentaphylla, 150-450 parts of fermented cordyceps sinensis powder, 50-250 parts of pine pollen, 50-250 parts of schisandra chinensis and a proper amount of starch, and the mass ratio of the compound shown in the formula (I) to the traditional Chinese medicine composition is (1-150): (1-150). # imgabs0 #

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a pharmaceutical composition and application thereof. Background Art

[0002] Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fibrotic interstitial pneumonia that occurs frequently in middle-aged and elderly people, with more men than women. The onset of the disease is insidious, with the main manifestations being active dyspnea, which progressively worsens and is often accompanied by dry cough. In severe cases, dyspnea and hypoxemia may occur, endangering the patient's life and health. At present, the pathogenesis of pulmonary fibrosis has not yet been clarified. The most effective clinical treatment is lung transplantation, but lung transplantation matching is difficult, expensive, and there will be rejection reactions after transplantation, so it is difficult to promote. Therefore, there is an urgent need to find a more effective and safe drug treatment method. Regarding drug treatment, Western medicine mainly uses immunosuppressants, adrenocortical hormones and other drugs for treatment, but the above drugs cannot achieve the ideal expected clinical effect.

[0003] In recent years, with the deepening of the understanding of the etiology and pathogenesis of IPF, it has been found that the pathological process of IPF includes repeated damage to epithelial cells, changes in immunity, vascular leakage and coagulation, abnormal wound healing, fibrosis, collagen accumulation and other links. Most of the IPF treatment drugs currently used in clinical practice cannot effectively treat multiple targets, and the effect is not ideal. Therefore, it is urgent to improve the clinical treatment effect by combining drugs targeting a single target or multiple targets. Traditional Chinese medicine treatment has the advantages of multiple pharmacodynamic sites and multiple therapeutic targets. Through research, it has been found that traditional Chinese medicine treatment of IPF has unique advantages over Western medicine treatment in improving the quality of life of patients and improving immunity. It also has the advantages of multiple targets, low toxicity and side effects, low drug dependence, and low price. It has broad prospects and can be used as a choice of clinical medication for IPF and support for drug research.

[0004] There is no related report in the prior art on the use of the pharmaceutical composition claimed in the present invention in preventing and treating pulmonary fibrosis such as idiopathic pulmonary fibrosis. Summary of the invention

[0005] Based on this, the present invention provides a pharmaceutical composition comprising

[0006] (a) a compound of formula (I); and

[0007]

[0008] (b) a Chinese medicine composition, wherein the raw materials of the Chinese medicine composition include, by weight: 50-250 parts of peach kernel, 300-700 parts of salvia miltiorrhiza, 200-600 parts of gynostemma pentaphyllum, 150-450 parts of fermented cordyceps powder, 50-250 parts of pine pollen, 50-250 parts of schisandra chinensis, and an appropriate amount of starch.

[0009] Alternatively, the Chinese medicine composition is prepared by adding the following Chinese medicine extracts in the following weight percentage ratios as the active ingredients, and adding auxiliary materials: 25-55% of Salvia miltiorrhiza extract, 12-40% of Gynostemma pentaphyllum extract, 5-12% of Schisandra chinensis alcohol extract, 12-30% of Cordyceps mycelium extract, 6-12% of pine pollen extract, 5-30% of peach kernel extract, and the rest are auxiliary materials;

[0010] The above-mentioned pharmaceutical compound preparation wherein each effective component is prepared by the following method:

[0011] Danshen extract: Take Danshen medicinal materials, crush them into granules, add about 12 times the amount of water, extract twice at 95℃±2, combine the extracts and concentrate for use. After testing, the water-soluble components of Danshen are mainly salvianolic acid B and sodium danshen. The residues are refluxed with about 70% ethanol twice, each time for about 1 hour, and the combined extracts are concentrated for use. After testing, the fat-soluble components of Danshen are mainly tanshinone. The combined concentrates are dried for use; Gynostemma pentaphyllum extract: Gynostemma pentaphyllum medicinal materials are soaked in about 75% ethanol, and high-frequency ultrasonic extraction is performed 3 times to obtain Gynostemma pentaphyllum extract. After testing, the main component is total saponin; Schisandra chinensis ethanol extract: Take Schisandra chinensis raw materials and add about 80% ethanol to reflux and extract twice. The extract is dried after recovering ethanol. The powder is crushed to obtain fine powder. After testing, the lignan components in Schisandra chinensis are biphenylcyclooctene-type lignan components, mainly Schisandraein A. The extraction rate of Schisandraein A is 0.3% to 2.0% by reflux extraction with about 80% ethanol. Cordyceps mycelium extract: take Cordyceps mycelium raw material and add about 75% ethanol to warm extract three times, combine the extract, recover ethanol, and dry the extract for later use. Pine pollen lipid extract: use about 70% ethanol to reflux heat extraction of pine pollen, recover ethanol from the extract, and dry the extract for later use. Peach kernel extract: the effective ingredient of the traditional Chinese medicine peach kernel is a water-soluble component. The water decoction method is adopted. The peach kernel is decocted twice with about 10 times the amount of water, and the extract is concentrated to an extract, and the extract is dried for later use.

[0012] The above-mentioned extracts are dried by vacuum drying or spray drying.

[0013] Wherein, the mass ratio between the compound of formula (I) and the traditional Chinese medicine composition is (1-150):(1-150).

[0014] Furthermore, the raw materials of the traditional Chinese medicine composition include, by weight: 100-200 parts of peach kernels, 500-600 parts of salvia miltiorrhiza, 300-500 parts of gynostemma pentaphyllum, 200-300 parts of fermented cordyceps powder, 100-200 parts of pine pollen, 100-200 parts of schisandra chinensis, and an appropriate amount of starch.

[0015] Furthermore, in parts by weight, the raw materials of the traditional Chinese medicine composition include: about 130 parts of peach kernel, about 530 parts of salvia miltiorrhiza, about 400 parts of gynostemma pentaphyllum, about 260 parts of fermented cordyceps powder, about 130 parts of pine pollen, about 130 parts of schisandra chinensis, and an appropriate amount of starch.

[0016] Further, the mass ratio between the compound of formula (I) and the Chinese medicine composition is (3-120): (8-120). Further, the mass ratio between the compound of formula (I) and the Chinese medicine composition is (1-5): (5-10). Further, the mass ratio between the compound of formula (I) and the Chinese medicine composition is about 3: about 8. Further, the mass ratio between the compound of formula (I) and the Chinese medicine composition is (2.85-3.15): (7.6-8.4).

[0017] Furthermore, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and treating diseases related to pulmonary fibrosis.

[0018] Furthermore, the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0019] Further, the drug is selected from one or more of the following: tyrosine kinase inhibitors, antacids, antioxidants, anti-connective tissue growth factor antibodies, lysophosphatidic acid inhibitors, pentraxin-2, JAK-STAT signaling pathway inhibitors, phosphodiesterase 4 inhibitors and TAS-115. Further, the tyrosine kinase inhibitor is nintedanib. Further, the antacid is a proton pump inhibitor and / or a histamine 2 receptor antagonist. Further, the antioxidant is N-acetylcysteine.

[0020] Further, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients. Further, the excipients are selected from one or more of the following: diluents, wetting agents, adhesives, disintegrants, inclusion agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, opacifiers and antioxidants. Further, the dosage form of the pharmaceutical composition is powder, tablet, drop pill, capsule, film, lozenge, granule, injection or oral solution. Further, the dosage form of the pharmaceutical composition is capsule or tablet. Further, the tablet is a dispersible tablet or a sustained-release tablet.

[0021] Furthermore, the preparation method of the Chinese medicine composition comprises the following steps:

[0022] (1) adding water to decocting Danshen, peach kernel and Gynostemma pentaphyllum weighed in weight ratio, combining the decoctions, taking the supernatant, concentrating it into an extract, cooling it, adding ethanol, precipitating it and filtering it; (2) weighing fermented Cordyceps sinensis powder and Schisandra chinensis chinensis, adding ethanol to reflux, cooling it, combining the ethanol solution, filtering it, concentrating and drying the filtrate; (3) weighing pine pollen, adding ethanol to warm immerse it, combining the extracts, concentrating and drying it into a dry paste; and (4) taking the above three dry pastes, mixing and grinding them, adding an appropriate amount of starch, and mixing them evenly to obtain the Chinese medicine composition.

[0023] According to another aspect of the present invention, there is provided a use of the above-mentioned pharmaceutical composition in the preparation of a medicament for preventing and treating diseases related to pulmonary fibrosis.

[0024] Furthermore, the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0025] Furthermore, the pharmaceutical composition inhibits inflammation and fibrosis-related factors, regulates CD4 + The differentiation direction of T cell subsets plays a role in preventing and treating pulmonary fibrosis.

[0026] According to another aspect of the present invention, there is provided a use of the above-mentioned pharmaceutical composition in the preparation of a medicament for downregulating the expression of cytokines selected from the group consisting of: IL-4, IL-6, IL-17, TNF-α, MMP-2, TIMP-1, CTGF, TGF-β1 and PDGF.

[0027] According to another aspect of the present invention, there is provided a use of the above pharmaceutical composition in the preparation of a medicament for up-regulating the expression of cytokines selected from the group consisting of: CD4 / CD8, IFN-γ, IL-2 and IL-10.

[0028] Beneficial effects of the present invention:

[0029] The pharmaceutical composition of the present invention comprising the compound of formula (I) and the traditional Chinese medicine composition can be used for preventing and treating idiopathic pulmonary fibrosis. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without exceeding the scope of protection required by the present invention.

[0031] Figure 1 This is a schematic diagram of the effect of different concentrations of TGF-β1 on the proliferation of MRC-5 cells. * Compared with the model group, P<0.05, **Represents comparison with the model group, P < 0.01.

[0032] Figure 2 This is a schematic diagram of the effects of different drug combinations on MRC-5 cell proliferation. * Compared with the model group, P<0.05, ** represents comparison with the model group, P < 0.01; ## Representative comparison with the Fuzheng Huayu + pirfenidone group (clinical dosage ratio 1:1), P < 0.01.

[0033] Figure 3 This is a schematic diagram of the effects of different ratios of pirfenidone and Fuzheng Huayu on the proliferation of MRC-5 cells. * Compared with the model group, P<0.05, ** represents comparison with the model group, P < 0.01; # Compared with the Fuzheng Huayu + pirfenidone (8:3) group, P<0.05, ## Representative comparison with the Fuzheng Huayu + pirfenidone (8:3) group, P < 0.01. DETAILED DESCRIPTION

[0034] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of the present invention.

[0035] Unless otherwise indicated, all technical and scientific terms and abbreviations used herein have the meanings commonly understood by those of ordinary skill in the field of the invention or the field in which the terms are used. Although any methods, conditions, substances or materials similar to or equivalent to those disclosed herein may be used in the practice of the present invention, preferred methods, conditions, substances or materials are described herein.

[0036] The present invention is intended to encompass all alternatives, variations and equivalents that may be included in the present invention as defined by the claims. Those skilled in the art will recognize many methods and materials similar or equivalent to those described herein that can be used in the practice of the present invention. The present invention is in no way limited to the methods and materials described.

[0037] As used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0038] In the present invention, the term "comprising" is synonymous with "including". As used herein, the terms "comprising", "including", "having", "containing" or any other variation thereof are intended to cover a non-exclusive inclusion. For example, a composition, process, method, article, or apparatus comprising the listed elements is not necessarily limited to only those elements, but may include other elements not expressly listed or inherent to such composition, process, method, article, or apparatus.

[0039] As described in the background technology section, there is no related report in the prior art on the use of the pharmaceutical composition claimed in the present invention in preventing and treating pulmonary fibrosis. In order to solve the above problems, the present invention provides a pharmaceutical composition, which comprises

[0040] (a) a compound of formula (I) (i.e., "pirfenidone"); and

[0041]

[0042] (b) a Chinese medicine composition, wherein the raw materials of the Chinese medicine composition include, by weight: 50-250 parts of peach kernel, 300-700 parts of salvia miltiorrhiza, 200-600 parts of gynostemma pentaphyllum, 150-450 parts of fermented cordyceps powder, 50-250 parts of pine pollen, 50-250 parts of schisandra chinensis, and an appropriate amount of starch.

[0043] Alternatively, the Chinese medicine composition is prepared by adding the following Chinese medicine extracts in the following weight percentage ratios as the active ingredients, and adding auxiliary materials: 25-55% of Salvia miltiorrhiza extract, 12-40% of Gynostemma pentaphyllum extract, 5-12% of Schisandra chinensis alcohol extract, 12-30% of Cordyceps mycelium extract, 6-12% of pine pollen extract, 5-30% of peach kernel extract, and the rest are auxiliary materials;

[0044] The above-mentioned pharmaceutical compound preparation wherein each effective component is prepared by the following method:

[0045] Danshen extract: Take Danshen medicinal materials, crush them into granules, add about 12 times the amount of water, extract twice at 95℃±2, combine the extracts and concentrate for use. After testing, the water-soluble components of Danshen are mainly salvianolic acid B and sodium danshen. The residues are refluxed with about 70% ethanol twice, each time for about 1 hour, and the combined extracts are concentrated for use. After testing, the fat-soluble components of Danshen are mainly tanshinone. The combined concentrates are dried for use; Gynostemma pentaphyllum extract: Gynostemma pentaphyllum medicinal materials are soaked in about 75% ethanol, and high-frequency ultrasonic extraction is performed 3 times to obtain Gynostemma pentaphyllum extract. After testing, the main component is total saponin; Schisandra chinensis ethanol extract: Take Schisandra chinensis raw materials and add about 80% ethanol to reflux and extract twice. The extract is dried after recovering ethanol. The powder is crushed to obtain fine powder. After testing, the lignan components in Schisandra chinensis are biphenylcyclooctene-type lignan components, mainly Schisandraein A. The extraction rate of Schisandraein A is 0.3% to 2.0% by reflux extraction with about 80% ethanol. Cordyceps mycelium extract: take Cordyceps mycelium raw material and add about 75% ethanol to warm extract three times, combine the extract, recover ethanol, and dry the extract for later use. Pine pollen lipid extract: use about 70% ethanol to reflux heat extraction of pine pollen, recover ethanol from the extract, and dry the extract for later use. Peach kernel extract: the effective ingredient of the traditional Chinese medicine peach kernel is a water-soluble component. The water decoction method is adopted. The peach kernel is decocted twice with about 10 times the amount of water, and the extract is concentrated to an extract, and the extract is dried for later use.

[0046] The above-mentioned extracts are dried by vacuum drying or spray drying.

[0047] Wherein, the mass ratio between the compound of formula (I) and the traditional Chinese medicine composition is (1-150):(1-150).

[0048] In the present invention, when mass ratio, weight part, weight percentage, time, pressure, equivalent, concentration or other value or parameter is expressed as a range, preferred range, or a range limited by a series of upper preferred values ​​and lower preferred values, it should be understood that all ranges formed by any pairing of any range upper limit or preferred value and any range lower limit or preferred value are specifically disclosed, regardless of whether the range is disclosed separately. For example, when the range "1 to 150" is disclosed, the described range should be interpreted as including the range "1 to 150", "1 to 120", "1 to 100", "1 to 80", "1 to 60", "1 to 40", "1 to 20", "1 to 10", etc. When a numerical range is described in this article, unless otherwise stated, the range is intended to include its end value and all integers and fractions within the range.

[0049] In a preferred embodiment, the raw materials of the traditional Chinese medicine composition include, by weight: 100-200 parts of peach kernel, 500-600 parts of salvia miltiorrhiza, 300-500 parts of gynostemma pentaphyllum, 200-300 parts of fermented cordyceps powder, 100-200 parts of pine pollen, 100-200 parts of schisandra chinensis, and an appropriate amount of starch.

[0050] In a preferred embodiment, the raw materials of the traditional Chinese medicine composition include, by weight: about 130 parts of peach kernel, about 530 parts of salvia miltiorrhiza, about 400 parts of gynostemma pentaphyllum, about 260 parts of fermented cordyceps powder, about 130 parts of pine pollen, about 130 parts of schisandra chinensis, and an appropriate amount of starch.

[0051] In the present invention, "about" refers to a value within a range of ±5% of a particular value. For example, “about 130” includes ±5% of 130, or from 123.5 to 136.5; “about 530” includes ±5% of 530, or from 503.5 to 556.5; “about 400” includes ±5% of 400, or from 380 to 420; “about 260” includes ±5% of 260, or from 247 to 273; “about 12” includes ±5% of 12, or from 11.4 to 12.6; “about 70” includes ±5% of 70, or from 66.5 to 73.5; “about 1” includes ±5% of 1, or from 0.95 to 1.05; “about 75” includes ±5% of 75, or from 71.25 to 78.75; “about 80” includes ±5% of 80, or from 76 to 84; “about 10” includes ±5% of 10, or from 9.5 to 10.5.

[0052] In a preferred embodiment, the mass ratio between the compound of formula (I) and the Chinese medicine composition is (3-120): (8-120). In a preferred embodiment, the mass ratio between the compound of formula (I) and the Chinese medicine composition is (1-5): (5-10). In a preferred embodiment, the mass ratio between the compound of formula (I) and the Chinese medicine composition is about 3: about 8. In a preferred embodiment, the mass ratio between the compound of formula (I) and the Chinese medicine composition is (2.85-3.15): (7.6-8.4).

[0053] In the present invention, "about" refers to a value within a range of ±5% of a specific value. For example, "about 3" includes ±5% of 3, or from 2.85 to 3.15; "about 8" includes ±5% of 8, or from 7.6 to 8.4.

[0054] In a preferred embodiment, the pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and treating diseases related to pulmonary fibrosis.

[0055] In the present invention, the term "prevention" includes "treatment" and "prevention", unless there is a specific description to the contrary. The terms "treatment" and "therapeutically" should be understood accordingly.

[0056] In the present invention, the term "treatment" includes alleviating, inhibiting or improving the symptoms or conditions of a disease; inhibiting the occurrence of complications; improving or preventing potential metabolic syndrome; inhibiting the occurrence of a disease or symptom, such as controlling the development of a disease or condition; alleviating a disease or symptom; reducing a disease or symptom; alleviating complications caused by a disease or symptom, or preventing or treating signs caused by a disease or symptom. As used herein, a pharmaceutical composition or a pharmaceutical preparation, after administration, can improve a disease, symptom or condition, especially improve its severity, delay the onset, slow the progression of the disease, or reduce the duration of the disease. Whether fixed or temporary administration, continuous or intermittent administration, can be attributed to or related to the administration.

[0057] In a preferred embodiment, the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0058] In a preferred embodiment, the drug is selected from one or more of the following: tyrosine kinase inhibitors, antacids, antioxidants, anti-connective tissue growth factor antibodies, lysophosphatidic acid inhibitors, pentraxin-2, JAK-STAT signaling pathway inhibitors, phosphodiesterase 4 inhibitors and TAS-115. In a preferred embodiment, the tyrosine kinase inhibitor is nintedanib. In a preferred embodiment, the antacid is a proton pump inhibitor and / or a histamine 2 receptor antagonist. In a preferred embodiment, the antioxidant is N-acetylcysteine.

[0059] In a preferred embodiment, the pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

[0060] In a preferred embodiment, the pharmaceutical preparation of the present invention contains at least one pharmaceutically acceptable excipient in an amount of 0.00001 to 50 wt.%, or 0.0001 to 10 wt.%, or 0.0001 to 5 wt.%, or 0.005 to 1 wt.%, or 0.1 to 20 wt.%, or 0.5 to 15 wt.%, or 1 to 5 wt.%, based on the weight of the pharmaceutical preparation.

[0061] In the present invention, the term "pharmaceutically acceptable" refers to a substance, such as a carrier or diluent, that does not abrogate the biological activity or properties of the compound and is relatively non-toxic, i.e., a substance that does not cause undesirable biological effects or interact in a deleterious manner with any of its components when administered to an individual.

[0062] In the present invention, the term "pharmaceutically acceptable excipient" refers to carriers and / or excipients that are pharmacologically and / or physiologically compatible with subjects and active ingredients (i.e., capable of inducing the desired therapeutic effect without causing any undesirable local or systemic effects), which are well known in the art (see, for example, Remington's Pharmaceutical Sciences. Edited by Gennaro AR, 19th ed. Pennsylvania: Mack Publishing Company, 1995).

[0063] In a preferred embodiment, the auxiliary material is selected from one or more of the following: diluent, wetting agent, binder, disintegrant, inclusion agent, flavoring agent, sustained-release agent, retention agent, lubricant, dispersant, plasticizer, opacifier and antioxidant.

[0064] Those skilled in the art will know how to select specific chemical substances within the scope of the above-mentioned auxiliary material category.For example, the diluent can be selected from one or more of the following: icing sugar, starch, compressible starch, lactose, dextrin, mannitol, sorbitol, microcrystalline cellulose, calcium sulfate and calcium carbonate.The wetting agent can be selected from one or more of the following: polyoxymethylene stearate, poloxamer, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene hydrogenated castor oil, polyoxyethylene alkyl ether, polysorbate such as polysorbate 80, cetyl alcohol, glycerol fatty acid ester (such as triacetin, glyceryl monostearate and the like), polyoxyethylene fatty acid ester, polyethylene glycol fatty acid ester, sodium lauryl sulfate, sorbitol fatty acid ester, sucrose fatty acid ester, polyoxyethylene ether, benzalkonium chloride, polyoxyethylene castor oil and docusate sodium.The adhesive can be selected from one or more of the following: polyvinyl pyrrolidone, hydroxypropyl cellulose, polyethylene glycol and methylcellulose. The disintegrant may be selected from one or more of the following: carboxymethylcellulose, carboxymethylcellulose calcium salt and sodium carboxymethylcellulose. The flavoring agent may be selected from one or more of the following: sorbitol, glucose, mannose, sucrose and lactose. The dispersant may be selected from one or more of the following: cross-linked sodium carboxymethylcellulose, sodium starch glycolate and pre-gelatinized corn starch. The plasticizer may be dibutyl sebacate and / or various citrates. The sustained-release agent may be selected from one or more of the following: sodium carboxymethylcellulose, low-substituted hydroxypropylcellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, hydroxyethylcellulose, gum arabic, gelatin and shellac. The antioxidant may be selected from one or more of the following: sodium bisulfite, sodium pyrosulfite, sodium sulfite and sodium thiosulfate. The lubricant may be selected from one or more of the following: calcium stearate, talc, magnesium stearate, stearic acid and colloidal silicon dioxide.

[0065] These excipients are preferably pharmaceutically inert, or may have a synergistic or additive effect to enhance the therapeutic activity of the pharmaceutical composition, and the above-mentioned excipients are only listed. The excipients actually used in the present invention are not limited to the above-mentioned excipients and can be adjusted according to actual conditions to achieve the effects of the present invention.

[0066] In a preferred embodiment, the dosage form of the pharmaceutical composition is a powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution. In a preferred embodiment, the dosage form of the pharmaceutical composition is a capsule or tablet. In a preferred embodiment, the tablet is a dispersible tablet or a sustained-release tablet.

[0067] In a preferred embodiment, the preparation method of the Chinese medicine composition comprises the following steps:

[0068] (1) adding water to decocting Danshen, peach kernel and Gynostemma pentaphyllum weighed in weight ratio, combining the decoctions, taking the supernatant, concentrating it into an extract, cooling it, adding ethanol, precipitating it and filtering it; (2) weighing fermented Cordyceps sinensis powder and Schisandra chinensis chinensis, adding ethanol to reflux, cooling it, combining the ethanol solution, filtering it, concentrating and drying the filtrate; (3) weighing pine pollen, adding ethanol to warm immerse it, combining the extracts, concentrating and drying it into a dry paste; and (4) taking the above three dry pastes, mixing and grinding them, adding an appropriate amount of starch, and mixing them evenly to obtain the Chinese medicine composition.

[0069] According to another aspect of the present invention, there is provided a use of the above-mentioned pharmaceutical composition in the preparation of a medicament for preventing and treating diseases related to pulmonary fibrosis.

[0070] According to another aspect of the present invention, there is provided a pharmaceutical composition for preventing and treating diseases related to pulmonary fibrosis.

[0071] According to another aspect of the present invention, a method for preventing and treating diseases related to pulmonary fibrosis is provided, comprising administering an effective amount of the above pharmaceutical composition to the subject.

[0072] In the present invention, the term "subject" is a mammal. The mammal may be a human, a non-human primate, a mouse, a rat, a dog, a cat, a horse or a cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects representing the pulmonary fibrosis model. Preferably, the subject is a human.

[0073] The "effective amount" of the pharmaceutical composition or preparation used in the present invention can obtain the desired treatment and / or prevention effect. The amount effective for this purpose will depend on, for example, the pharmaceutical composition, the mode of administration, the stage and severity of the disease to be treated, the individual's body weight and overall health, and the judgment of the prescribing physician. The dosage can be given once a week, or once every two days or every day, or even several times a day. The dosage unit can be given in a short period of time (e.g., weeks to months) or longer time periods (months to years).

[0074] In a preferred embodiment, the pulmonary fibrosis is idiopathic pulmonary fibrosis.

[0075] In a preferred embodiment, the pharmaceutical composition inhibits inflammation and fibrosis related factors, regulates CD4 + The differentiation direction of T cell subsets plays a role in preventing and treating pulmonary fibrosis.

[0076] According to another aspect of the present invention, there is provided a use of the above-mentioned pharmaceutical composition in the preparation of a medicament for downregulating the expression of cytokines selected from the group consisting of: IL-4, IL-6, IL-17, TNF-α, MMP-2, TIMP-1, CTGF, TGF-β1 and PDGF.

[0077] According to another aspect of the present invention, there is provided a use of the above pharmaceutical composition in the preparation of a medicament for up-regulating the expression of cytokines selected from the group consisting of: CD4 / CD8, IFN-γ, IL-2 and IL-10.

[0078] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions or conditions recommended by the manufacturer.

[0079] Unless otherwise defined, all professional and scientific terms used herein have the same meanings as those familiar to those skilled in the art. In addition, any methods and materials similar or equivalent to those described herein can be applied to the methods of the present invention. The preferred implementation methods and materials described herein are for demonstration purposes only.

[0080] The above features mentioned in the present invention or the features mentioned in the embodiments can be combined in any way. All the features disclosed in this patent specification can be used in combination with any combination form, and each feature disclosed in the specification can be replaced by any alternative feature that can provide the same, equal or similar purpose. Therefore, unless otherwise specified, the disclosed features are only general examples of equal or similar features.

[0081] Example

[0082] Example 1 Cell in vitro screening test

[0083] 1. Test materials

[0084] MRC-5 cells (KeyGEN) were purchased from Wuhan Punosai Life Science Co., Ltd. (Wuhan, China); fetal bovine serum, MEM, non-essential amino acids, penicillin / streptomycin double antibody solution (Gibco); CCK8 (biosharp); PAGE gel rapid preparation kit 10% (Yazyme); SDS-PAGE electrophoresis instrument (Bio-Road); reverse transcription kit, sybr green (TOYOBO); Trizol (Invitrogen); antibodies α-SMA, Vimentin, E-cadherin, collagen I (CST); TGF-β1 item number: CYT-716, ProsPec. The tested Chinese medicine composition was provided by Shanghai Huanghai Pharmaceutical Co., Ltd. and prepared with reference to CN99113887.2 and CN200610106720.1 (the contents of the above two patents are introduced into this patent in full text).

[0085] 2. Test methods

[0086] 2.1. Drug combination screening and determination

[0087] 2.1.1. Determination method

[0088] 2.1.1.1. Screening method for optimal induction concentration of TGF-β1

[0089] MRC-5 cells were cultured in MEM containing 10% fetal bovine serum, 1% non-essential amino acids, 100U / mL penicillin and 100μg / mL streptomycin at 37°C and 5% CO2 incubator. MRC-5 cells were seeded in 96-well plates and cultured at 37°C for 24 hours. Different concentrations of TGF-β1 (0ng / mL, 0.625ng / mL, 1.25ng / mL, 2.5ng / mL, 5ng / mL, 10ng / mL, 20ng / mL, 40ng / mL, 80ng / mL) were added. After 48 hours of culture, CCK-8 was added and incubated for 2 hours, and the absorbance (A) value was read. The cell survival rate was calculated according to the following formula:

[0090] Cell survival rate (%) = (A 治疗 –A 空白 ) / (A 对照 –A 空白 )×100

[0091] 2.1.1.2. Screening methods for different drug combinations

[0092] Cells in logarithmic growth phase were obtained and trypsinized to obtain single cell suspension. The suspension was inoculated into a 96-well plate and the number of cells per well was controlled to be 1×10 3 The volume was 100 μL, and the area around the inoculated cell wells was filled with 1×PBS. When about 80% of the cells adhered to the wall, they were randomly divided into a blank group, a model group, and different anti-fibrosis drug combinations (pirfenidone; Fuzheng Huayu group; Fuzheng Huayu + pirfenidone; Fuzheng Huayu + Anluo Huaxian Pills; Fuzheng Huayu + Nintedanib; Fufang Biejia Ruangan Tablets + pirfenidone) for treatment, with 6 replicate wells in each group.

[0093] Drug preparation was designed according to 1 times or 1:1 of the clinical dose of the drug, and the drug concentrations added to each well were: (18 μg / mL of pirfenidone; 48 μg / mL of the Fuzheng Huayu group; Fuzheng Huayu: pirfenidone = (8:3) (48 μg / mL: 18 μg / mL); Fuzheng Huayu: Fufang Biejia Ruangan Tablets = 4:5 (48 μg / mL: 60 μg / mL); Fuzheng Huayu: Nintedanib = 16:1 (48 μg / mL: 3 μg / mL); Fufang Biejia Ruangan Tablets: pirfenidone = 30:9 (60 μg / mL: 18 μg / mL)

[0094] About 24 hours after drug treatment, when about 90% of the cells were attached to the wall, 10 μL of CCK-8 solution was added to each well, and the cells were cultured at 37°C for 2 hours. The absorbance of each well at 450 nm was read using an enzyme reader.

[0095] Cell survival rate (%) = (A 治疗 –A 空白 ) / (A 对照 –A 空白 )×100

[0096] 2.1.2 Drug screening results

[0097] 2.1.2.1 Determination of the induction concentration of TGF-β1

[0098] TGF-β1 can promote the transformation of MRC-5 cells into myofibroblasts. The optimal TGF-β1 stimulation condition was determined by the cell proliferation rate. Compared with the control group, the differences in TGF-β1 at 2.5 ng / mL, 5 ng / mL, 10 ng / mL, and 20 ng / mL were statistically significant (P<0.01). The optimal stimulation concentration of TGF-β1 was determined to be 5 ng / mL. Figure 1 And as shown in Table 1.

[0099] Table 1 Effects of different concentrations of TGF-β1 on the proliferation of MRC-5 cells

[0100] Group Is there a difference P-value 0vs.0.625 no 0.9994 0vs.1.25 no 0.9946 0vs.2.5 yes <0.0001 0vs.5 yes <0.0001 0vs.10 yes <0.0001 0vs.20 yes 0.0136 0vs.40 yes 0.0196 0vs.80 no 0.9946

[0101] 2.1.2.2 Screening results of different drug combinations

[0102] Through the use of drug combinations to improve the effect of TGF-β1 on cell proliferation, it was found that the pirfenidone, Fuzheng Huayu and other drug groups had significant inhibitory effects on the proliferation of MRC-5 cells induced by TGF-β1 compared with the model group (P<0.01, P<0.05); in the combination, the Fuzheng Huayu + pirfenidone combination had a more significant inhibitory effect on the inhibitory effect of TGF-β1 cell proliferation (P<0.01), which was better than the pirfenidone and Fuzheng Huayu groups alone. Therefore, the Fuzheng Huayu + pirfenidone combination is the best and most effective drug combination. The results are as follows Figure 2 And as shown in Table 2.

[0103] Table 2 Effects of different drug combinations on MRC-5 cell proliferation

[0104]

[0105]

[0106] Note: Different drug combinations were compared based on a 1:1 ratio of the clinical daily dose of the drugs.

[0107] 2.2. Drug combination ratio screening

[0108] Through the use of drug compositions to improve the effect of TGF-β1 on cell proliferation, it was found that the combination of Fuzheng Huayu (FZHY) + pirfenidone (BFNT) had a gradually obvious inhibitory effect on the inhibitory effect of TGF-β1 on cell proliferation. Therefore, the combination of pirfenidone and Fuzheng Huayu (pirfenidone: Fuzheng Huayu = 3:120, 3:20, 3:8, 7.5:8, 15:8, 30:8, 120:8) was used for further experiments.

[0109] Experimental method: Take cells in logarithmic growth phase, digest them with trypsin to obtain single cell suspension, inoculate the suspension into a 96-well plate, and control the number of cells in each well to 1×10 4 , volume 100 μL, the area around the inoculated cell wells was filled with 1×PBS. When about 80% of the cells adhered to the wall, they were randomly divided into a blank group, a model group, and different anti-fibrotic drug combinations (pirfenidone: Fuzheng Huayu = 3:120, 3:20, 3:8, 7.5:8, 15:8, 30:8, 120:8) for treatment. Each group was set up with 6 replicate wells, and each well was added with drug solution, the drug concentration was 10 μg / mL, and about 24 hours after drug treatment, 10 μL CCK-8 solution was added, and cultured at 37°C for 2 hours, and the absorbance of each well at 450nm was read with an enzyme marker.

[0110] Cell survival rate (%) = (A 治疗 –A 空白) / (A 对照 –A 空白 )×100

[0111] The experimental results are as follows Figure 3 As shown, pirfenidone: Fuzheng Huayu (3-120): (8-120) had a significant inhibitory effect on TGF-β1 cells (P < 0.05, P < 0.01), among which pirfenidone: Fuzheng Huayu = (3:8) had a more significant inhibitory effect on the TGF-β1 cell proliferation inhibition (P < 0.01); pirfenidone: Fuzheng Huayu = (3:8) had a more significant inhibitory effect on the TGF-β1 cell proliferation inhibition in different proportions of each combination, with a significant difference (P < 0.05, P < 0.01).

[0112] 2.3. Further comparison of drug combination ratio screening

[0113] 2.3.1. Histobiochemical comparison of drug ratios

[0114] MRC-5 cells were cultured in a DMEM medium containing 10% FBS in a 37°C, 5% CO2 incubator. When the cells reached 80% density, different ratios of combined drugs (pirfenidone: Fuzheng Huayu 3:8, 3:20, 7.5:8) were used, and each well was added with a drug solution at a drug concentration of 10μg / mL for 24h, and then stimulated with 5ng / mL TGF-β1 stimulator for 2h. The expression of ERK and inflammation-related pathway proteins in cells was detected by Western blot. After the culture was completed, RIPA lysis buffer was added to lyse the cells, centrifuged (4°C, 10000r / min, 10min), the supernatant was collected, the total protein concentration in the supernatant was determined by BCA method, the loading buffer was added, and the protein was denatured by boiling in a 96°C metal bath for 10min. Protein components were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (voltage 120V, time 2h), and the protein was transferred to polyvinylidene difluoride membrane by wet transfer method (voltage 80V, time 2h), blocked with 5% skim milk powder for 2h, and washed with PBST three times; α-SMA, collagen I, ERK1 / 2, p-ERK1 / 2 and GAPDH primary antibodies (dilution ratio of 1:1000) were added, incubated at 4℃ overnight, and washed with PBST three times; secondary antibodies (dilution ratio of 1:10000) were added and incubated at room temperature for 2h. Chemiluminescence imaging system was used for exposure and photography, and Image-Pro Plus 6.0 software was used to analyze the gray value of protein bands. The expression level of p-ERK1 / 2 protein was expressed by the ratio of the gray value of p-ERK1 / 2 protein band to that of ERK1 / 2 protein band, and the expression level of the target protein was expressed by the ratio of the gray value of the target protein band to that of the internal reference protein (GAPDH).

[0115] 2.3.2 Comparison results

[0116] Effects of pirfenidone + Fuzheng Huayu on the expression of α-SMA, collagen I and pERK1 / 2 proteins in MRC-5 cells. Compared with the blank group, the expression of α-SMA, collagen I and p-ERK1 / 2 proteins in the model group were significantly upregulated (P < 0.01); compared with the model group, the above proteins in the pirfenidone + Fuzheng Huayu different ratio groups were significantly downregulated (P < 0.05, P < 0.01). The results are shown in Tables 3 to 6.

[0117] Table 3 Results of the expression levels of α-SMA, collagen I and pERK1 / 2 proteins in each group of cells1 (`x±s, n=6)

[0118]

[0119] Table 4 Results of the expression levels of α-SMA, collagen I and pERK1 / 2 proteins in each group of cells2 (`x±s, n=6)

[0120]

[0121] Table 5 Results of the expression levels of α-SMA, collagen I and pERK1 / 2 proteins in each group of cells 3 (`x±s, n=6)

[0122]

[0123] Table 6 Results of the expression levels of α-SMA, collagen I and pERK1 / 2 proteins in each group of cells 4 (`x±s, n=6)

[0124]

[0125] Example 2 Animal Test

[0126] 1. Test materials

[0127] Microplate reader (Thermo, VARIOSKAN FLASH), Bleomycin (MCE (MedChemExpress LLC)), lipid peroxide (LPO) kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), hydroxyproline (Hyp) assay kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), malondialdehyde (MDA) assay kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), myeloperoxidase (MPO) kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), total superoxide dismutase (SOD) kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), IL-1β Elisa kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), TIMP-1 Elisa kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), MMP2 kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), PDGF Elisa kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), TNF-α Elisa kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.), TGF-β Elisa kit (Nanjing Jiancheng Haihao Biotechnology Co., Ltd.).

[0128] 2. Purpose of the test

[0129] In this study, the pharmacodynamic effects of FZHY and pirfenidone combined with bleomycin-induced pulmonary fibrosis in rats were determined in different mechanisms.

[0130] 3. Bleomycin-induced pulmonary fibrosis model in rats

[0131] The day before modeling of this experiment, the animals were randomly divided into 6 groups according to their body weight, as shown in Table 7.

[0132] Table 7 Animal grouping comparison information

[0133]

[0134]

[0135] 4. Modeling

[0136] 100 SD male rats were randomly divided into 8 groups, 10 to 15 rats in each group, and divided into blank control group and model group. After anesthesia, the rats were fixed on the operating table. After the neck was disinfected with alcohol cotton, the trachea was exposed layer by layer, and the needle was inserted into the heart end through the gap of the tracheal cartilage ring. After the injection of 3.5 mg / kg of bleomycin, the animals were quickly erected and then rotated for a moment to make the solution evenly distributed in the lungs, and the incision was sutured. Only the trachea was exposed in the sham operation group. After the animals woke up, they were randomly divided into 3 drug administration groups (FZHY combined with medication 1, 2, 3), 3 positive drug groups, and a model group. FZHY was given 7 days after modeling, and a total of 21 times were given. The model group and the sham operation group were given the corresponding solvent.

[0137] 5. Treatment groups of rats

[0138] The animals were divided into a blank group, a model group, a positive drug group (FZHY, pirfenidone, dexamethasone), and FZHY and pirfenidone combined drug groups 1, 2, and 3. The samples were collected after 28 days, and the samples were collected after the number of days of testing and anesthesia and sacrifice. Detection indicators: serum peroxidase (MPO) detection, lung perfusion fluid: alveolar perfusion fluid TGF-β1, TNF-α, lung tissue pathology detection (MASSON).

[0139] 6. Test results

[0140] The results showed that the combined drug groups 1, 2, and 3 had significant differences in MPO, TGF-β1, TNF-α, and MASSON staining compared with the model group (P < 0.01, P < 0.05), indicating that the combined drug treatment had a significant effect in improving the pulmonary fibrosis model, and the effect was better than that of pirfenidone and Fuzheng Huayu alone.

[0141] Example 3 Clinical Trial

[0142] 1. Case selection

[0143] 1.1. Source of cases

[0144] This study collected 80 patients with idiopathic pulmonary fibrosis (IPF) diagnosed as "pulmonary consumptive-Qi deficiency and blood stasis syndrome" in the wards and outpatient clinics of Baoshan Hospital Affiliated to Shanghai University of Traditional Chinese Medicine from December 27, 2021 to March 1, 2023 as research subjects. The 80 IPF patients who met the inclusion criteria were randomly divided into a treatment group and a control group. The treatment group was treated with pirfenidone combined with a Chinese medicine composition, and the control group was treated with pirfenidone combined with a placebo. All patients included in this study signed written informed consent.

[0145] 1.2 Diagnostic criteria

[0146] 1.2.1 Western medicine diagnostic criteria

[0147] Refer to the relevant diagnostic criteria in the "Evidence-Based Guidelines for the Diagnosis and Treatment of Idiopathic Pulmonary Fibrosis (2011 Edition)".

[0148] 1.2.2 TCM diagnostic criteria

[0149] Formulated with reference to "Chinese Internal Medicine" (Edited by Zhou Zhongying, China Traditional Chinese Medicine Press, Beijing, 2003) and "Chinese Medicine Diagnosis and Treatment Program for Pulmonary Consumption (Pulmonary Interstitial Fibrosis) (Trial)" in 2012. Patients who meet the Chinese medicine syndrome diagnosis belong to the "pulmonary consumption" syndrome of qi deficiency and blood stasis. Main symptoms: shortness of breath and wheezing, which is more obvious after activity, cough and sputum. Secondary symptoms: chest pain and chest tightness, fatigue, less sputum or dry cough without sputum, cough with wheezing, or cyanosis of the lips, nails, clubbing, dull skin, or accompanied by aversion to wind and cold, dry mouth and thirst, dizziness and tinnitus, soreness of waist and knees, and edema of lower limbs. Tongue: The tongue is dull or has petechiae and ecchymosis, and the tongue coating is mostly thin and white. Pulse: deep and fine or astringent. With the main symptoms and more than 2 secondary symptoms, the diagnosis can be made by referring to the tongue and pulse.

[0150] 1.3 Inclusion criteria

[0151] ① Meet the Western medical diagnostic criteria for IPF; ② Meet the TCM syndrome differentiation criteria for qi deficiency and blood stasis in the "pulmonary consumption" syndrome; ③ Age 50 to 89 years old; ④ Sign the informed consent form.

[0152] 1.4 Exclusion criteria

[0153] ① Patients with acute IPF attack or patients with FVC <50% in lung function test or extremely critical patients with obvious lung infection requiring anti-infection treatment; ② Patients with combined severe infection, severe hypertension, coronary heart disease, heart failure, obvious abnormal liver and kidney function, etc.; ③ Patients with chronic cough and wheezing caused by tuberculosis, fungi, tumors, silicosis, irritating gases, allergies, etc. confirmed by examination; ④ Disabled patients stipulated by law (blind, deaf, dumb, intellectual disability, mental disorder, etc.); ⑤ Pregnant or lactating women and those known to be allergic to the components of this drug; ⑥ Observers who participated in clinical trials of other drugs in the past 3 months and affected the effect indicators of this study.

[0154] 2. Research Methods

[0155] 2.1 Drug treatment regimen

[0156] Control group: Patients routinely took pirfenidone capsules (manufacturer: Beijing Contini Pharmaceutical Co., Ltd., approval number: National Medicine Standard H20133376), specification: 100 mg per capsule. Usage and dosage: Oral, 4 capsules at a time, 3 times a day, oral with warm water, 3 months as a course of treatment. Simultaneously take a simulated placebo: 5 capsules at a time, 3 times a day, oral with warm water, half an hour after a meal, 3 months as a course of treatment.

[0157] Treatment group: After admission, the patients took pirfenidone in the same dosage and usage as the control group, and were given a Chinese medicine composition (manufacturer: Shanghai Huanghai Pharmaceutical Co., Ltd., approval number: National Medicine Standard Z20050546, prepared with reference to CN99113887.2 and CN200610106720.1, the contents of the above two patents are introduced into this patent in full text), specification: 0.4g per tablet. Usage and dosage: oral, 5 tablets at a time, 3 times a day, oral with warm water, half an hour after a meal, 3 months as a course of treatment.

[0158] 2.2. General information collection

[0159] 2.2.1. General record items

[0160] Gender, age, patient randomization code, drug number, patient address, contact information, observing physician, subject name, trial start date, etc.

[0161] 2.2.2 Biological indicators

[0162] Including height, weight and vital signs (body temperature, resting heart rate, respiration, blood pressure), tongue coating, and pulse.

[0163] 2.3 Observation indicators

[0164] 2.3.1. Main efficacy indicators

[0165] (1) TCM syndrome score

[0166] The evaluation criteria were formulated in accordance with the "Guidelines for Clinical Research of New Chinese Medicines (Trial)" (2002 edition). The TCM clinical syndrome scores of all enrolled patients before and after treatment were comprehensively evaluated by two quality control personnel at the level of deputy chief physicians. Including the main symptom scores: wheezing, shortness of breath, sputum, cough; secondary or concurrent symptom scores: chest tightness, sputum color, fever, itchy throat, aversion to wind, sweating, fatigue, palpitations, purple lips and nails, clubbing, tongue coating, pulse, Velcros rales. The difference in the total score of TCM clinical symptoms before and after treatment (0 and 3 months) was compared.

[0167] 2.3.2. Secondary efficacy indicators

[0168] (1) Dyspnea Index Score

[0169] Referring to Table 8, the Borg scale scores of all enrolled patients before and after treatment were comprehensively evaluated by two quality control personnel at the associate chief physician level.

[0170] Table 8 Borg dyspnea index score table

[0171] Points Performance 1 No breathlessness or fatigue at all 2 Very mild shortness of breath or fatigue, barely noticeable 3 Very mild breathing difficulty or fatigue 4 Mild shortness of breath or fatigue 5 Moderate dyspnea or fatigue 6 Slightly severe shortness of breath or fatigue 7 Severe breathing difficulties or fatigue 8 Very severe breathing difficulty or fatigue 9 Very severe breathing difficulty or fatigue 10 Extreme shortness of breath or fatigue, reaching the limit

[0172] (2) St. George's Respiratory Questionnaire

[0173] The St. George's Respiratory Questionnaire (SGRQ) score includes three parts: symptoms, activity ability and disease impact. The scores of the subjects before and after treatment were recorded and statistically analyzed.

[0174] (3) Chest high-resolution CT (HRCT) results

[0175] The evaluation criteria were formulated with reference to the literature, and the chest HRCT scores before and after treatment (0 and 3 months) were compared. The specific method is as follows:

[0176] A 64-row spiral CT machine was used for thin-layer scanning. HRCT images were read and scored by three chest radiologists (deputy director / director-attending physician-resident physician), and the average value was finally taken. HRCT scoring of the enrolled patients was performed according to the Muller method: the aortic arch level, tracheal bifurcation level, and 1 cm level above the diaphragm were selected, and the ground-glass changes, grid-like changes, and honeycomb-like changes were scored separately according to Tables 9 and 10. The total HRCT score was obtained by adding up the scores of the three changes.

[0177] Table 9 HRCT imaging feature scoring table (points)

[0178] Imaging features Slight attenuation Ground glass opacity Ground glass opacity with DB Reticular shadow Net Shadow DB Honeycomb Shadow score 1 2 3 4 5 6

[0179] Table 10 HRCT lesion area scoring table (points)

[0180] area(%) 0<IS≤5 5<IS≤25 25<IS≤50 50<IS≤75 75<IS≤100 score 1 2 3 4 5

[0181] (4) Lung function

[0182] The evaluation of pulmonary function efficacy was made according to references, and the vital capacity (VC), forced vital capacity (FVC) and diffusion capacity (DLCO) were compared before and after treatment (0 and 3 months).

[0183] (5) Inflammation-related cytokines and immune function detection

[0184] Before and after treatment, 2 mL of anticoagulated peripheral blood was collected from each subject, and the blood was naturally precipitated and centrifuged at 4°C to obtain the upper serum, which was stored at -80°C for later use. Flow cytometry was used to measure immune indicators such as CD4 / CD8, and ELISA was used to measure the expression of immune-related cytokines IFN-γ, TNF-α, IL-2, IL-4, IL-6, IL-10, and IL-17.

[0185] (6) Detection of fibrosis-related factors

[0186] The levels of serum TGF-β1, matrix metalloproteinase (MMP2), matrix metalloproteinase inhibitor (TIMP-1), connective tissue growth factor (CTGF), and platelet-derived growth factor (PDGF) before and after treatment were detected by ELISA method.

[0187] (7) Clinical efficacy observation

[0188] The evaluation criteria for TCM syndrome efficacy refer to the "Guidelines for Clinical Research of New Chinese Medicines (Trial)" (2002 Edition). Two quality control personnel at the level of associate chief physicians will evaluate the total score of TCM clinical syndromes of all enrolled patients before and after treatment (0 and 3 months). The evaluation criteria are divided into clinical cure (symptom score reduction ≥90%), marked effect (symptom score reduction ≥70%), effective (symptom score reduction ≥30%), and ineffective (symptom score reduction less than 30%).

[0189] 2.3.3. Adverse reaction observation

[0190] The control group and the treatment group were observed for abnormalities in liver and kidney function, coagulation mechanism disorders, drug allergies, and dropouts before and after treatment.

[0191] 3. Data Statistical Analysis

[0192] All data analyses were performed using SPSS 23.0 statistical software. Measurement data that met normal distribution and variance homogeneity were expressed as mean ± standard deviation (x ± s) and were tested by t test; measurement data that did not meet normal distribution and variance homogeneity were expressed as median and upper and lower interquartile range M (P25, P75), and non-parametric tests were used; the comparison of rank data between the two groups was performed by Mann-Whitney U test of two independent samples; count data were expressed as frequency and tested by non-parametric tests. The comparison between the two groups before and after treatment was performed by t test or non-parametric test. P < 0.05 indicated statistically significant differences.

[0193] 4. Results

[0194] During the clinical observation, the patients' conditions were relatively stable. No patients were hospitalized due to acute exacerbation within 3 months, and no special treatment was given. There was no significant change in the TCM syndrome type, and Qi deficiency and blood stasis were still the main syndrome types. There was no dropout in the treatment group and the control group. Finally, a total of 80 patients completed this clinical trial, with 40 cases in the treatment group and 40 cases in the control group.

[0195] 4.1 Impact on major clinical symptoms and total symptom score

[0196] As can be seen from Table 11, the improvement in the total symptom score in both groups was downward compared with before treatment (P<0.05), but the improvement effect of the Chinese medicine combination combined with pirfenidone treatment group on the total symptom score was significantly better than that in the control group, and the difference was statistically significant (P<0.05).

[0197] Table 11 Comparison of changes in main symptoms and total symptom scores between the two groups before and after treatment (M (P25, P75, points)

[0198]

[0199] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0200] 4.2 Effects on lung function (pulmonary ventilation function and diffusion function)

[0201] As shown in Table 12, compared with before treatment, the treatment group had significant improvements in VC%, FVC%, and DLCO% (P<0.05), while the control group had no significant improvement in VC%, FVC%, and DLCO% (P>0.05). Compared with the control group, the improvement in DLCO% in the treatment group was significantly better than that in the control group, and the difference was statistically significant (P<0.05).

[0202] Table 12 Comparison of lung function changes between the two groups before and after treatment ( % )

[0203]

[0204]

[0205] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0206] 4.3 Impact on Chest HRCT

[0207] As shown in Table 13, there was no significant difference in the HRCT score change before and after treatment in the control group (P>0.05), and the HRCT of the treatment group decreased after treatment compared with before treatment, and there was a statistical difference (P<0.05), but there was no statistically significant difference between the two groups after treatment (P>0.05).

[0208] Table 13 Comparison of HRCT scores between the two groups before and after treatment (M (P25, P75), points)

[0209]

[0210] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0211] 4.4 Changes in quality of life before and after treatment

[0212] As shown in Table 14, the scores of symptoms, activities and impacts in the treatment group decreased significantly compared with those before treatment (P<0.05); the scores of the control group did not decrease significantly compared with those before treatment (P>0.05). After treatment, the decreases of symptoms and impacts in the treatment group were more obvious than those in the control group, with significant differences (P<0.05).

[0213] Table 14 Comparison of changes in quality of life scores between the two groups before and after treatment ( point)

[0214] Group Symptoms section Activities Impact Control group Before treatment 51.03±7.03 43.15±2.23 39.43±3.30 (n＝40) After treatment 49.88±5.10 42.95±2.01 38.15±2.74 Treatment Group Before treatment 50.52±6.48 46.19±5.95 43.72±7.30 (n＝40) After treatment <![CDATA[46.37±5.39 *# ]]> <![CDATA[44.05±4.46 * ]]> <![CDATA[40.35±5.83 *# ]]>

[0215] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0216] 4.5 Impact on Dyspnea Index

[0217] There was no significant improvement in the dyspnea index score of the control group before and after treatment (P>0.05), while the dyspnea index score of the treatment group was significantly improved compared with before treatment (P<0.05), and there was a significant difference in the dyspnea index score between the two groups after treatment (P<0.05) (as shown in Table 15).

[0218] Table 15 Comparison of Borg dyspnea scores before and after treatment between the two groups (M (P25, P75), points)

[0219] n Before treatment After treatment Control group 40 3(2，3.75) 3(2，3) Treatment Group 40 2(2，3) <![CDATA[2(1,2.75) *# ]]>

[0220] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0221] 4.6 Effects on immune function and immune-related inflammatory cytokines

[0222] As shown in Table 16, compared with before treatment, the expression levels of CD4 / CD8 in both groups were increased after treatment, but there was a significant difference in the treatment group (P<0.05), while there was no significant difference in the control group (P>0.05). Comparing the two groups, the upregulation of CD4 / CD8 in the treatment group was significantly better than that in the control group.

[0223] As can be seen from Tables 17 and 18, compared with before treatment, the control group only had a significant up-regulation effect on IFN-γ (P<0.05); while the treatment group could significantly down-regulate the expression of pro-inflammatory cytokines IL-4, IL-6, IL-17, and TNF-α (P<0.05), and up-regulate the expression of anti-inflammatory factors IFN-γ, IL-2, and IL-10 (P<0.05); among them, the regulatory effect on IL-4, IL-6, IL-17, and IL-10 was significantly different from that of the control group (P<0.05), and there was no significant difference in the regulatory effect on other immune-related cytokines (P>0.05).

[0224] Table 16 Comparison of CD4 / CD8 expression between the two groups before and after treatment (M (P25, P75))

[0225] n Before treatment After treatment Control group 40 1.33(0.98，1.72) 1.68(1.54，2.18) Treatment Group 40 1.36(0.98，1.97) <![CDATA[2.40(1.46,3.20) *# ]]>

[0226] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0227] Table 17 Comparison of immune-related inflammatory cytokine expression between the two groups before and after treatment (M (P25, P75), pg / ml)

[0228]

[0229] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0230] Table 18 Comparison of immune-related inflammatory cytokine expression between the two groups before and after treatment (M (P25, P75), pg / ml)

[0231]

[0232] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0233] 4.7 Effects on fibrosis-related cytokines

[0234] As can be seen from Tables 19 and 20, after treatment, both the treatment group and the control group could down-regulate the expression of MMP-2, TIMP-1, CTGF, PDGF, and TGF-β1 in the patients' peripheral blood (P<0.05), and the down-regulation effect of the treatment group was more obvious than that of the control group (P<0.05).

[0235] Table 19 Comparison of the expression of MMP-2, TIMP-1 and CTGF before and after treatment between the two groups

[0236]

[0237] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0238] Table 20 Comparison of PDGF and TGF-β1 expression between the two groups before and after treatment ( ng / ml)

[0239] Group PDGF TGF-β1 Control group Before treatment 11.07±0.31 2.08±0.05 (n＝40) After treatment <![CDATA[9.67±0.23 * ]]> <![CDATA[1.65±0.05 * ]]> Treatment Group Before treatment 12.62±0.32 2.13±0.36 (n＝40) After treatment <![CDATA[8.23±0.20 *# ]]> <![CDATA[1.42±0.03 *# ]]>

[0240] Note: “*” means comparison with the group before treatment, P<0.05; “#” means comparison with the control group after treatment, P<0.05.

[0241] 4.8 Correlation Analysis

[0242] This study used Spearman correlation analysis to analyze the correlation between the difference in the total clinical score of the treatment group before and after treatment (after treatment - before treatment) and the difference in related detection indicators. It was found that the difference in the total clinical symptom score of the treatment group was negatively correlated with the difference in CD4 / CD8, DLCO%, and IL-2 levels, and positively correlated with the difference in TGF-β expression, and had no significant correlation with other detection indicators (P>0.05) (as shown in Table 21).

[0243] Table 21 Correlation analysis between the difference in total clinical scores before and after treatment and the difference in each test index in the treatment group

[0244]

[0245]

[0246] 4.9 Clinical efficacy observation

[0247] After treatment, 1 patient in the control group was clinically cured, 4 patients were markedly effective, 25 patients were effective, 9 patients were ineffective, and the total effective rate of the control group was 77.5%; 4 patients in the treatment group were clinically cured, 7 patients were markedly effective, 26 patients were effective, 3 patients were ineffective, and the total effective rate of the treatment group was 90%. The difference between the two groups was significant (P<0.05) (as shown in Table 22).

[0248] Table 22 Comparison of the effective rate of treatment between the two groups

[0249]

[0250] Note: According to non-parametric test, P < 0.05. There is a significant statistical difference in the effective rate between the two groups.

[0251] The embodiments of the present invention are described in detail above. Specific examples are used herein to illustrate the principles and implementation methods of the present invention. The description of the above embodiments is only used to help understand the method of the present invention and its core idea. At the same time, changes or deformations made by those skilled in the art based on the ideas of the present invention, the specific implementation methods and application scope of the present invention, all belong to the scope of protection of the present invention. In summary, the content of this specification should not be understood as limiting the present invention.

Claims

1. A pharmaceutical composition, characterized in that The pharmaceutical composition comprises (a) a compound of formula (I); and (b) a Chinese medicine composition, wherein the raw materials of the Chinese medicine composition include, by weight: 50-250 parts of peach kernel, 300-700 parts of salvia miltiorrhiza, 200-600 parts of gynostemma pentaphyllum, 150-450 parts of fermented cordyceps powder, 50-250 parts of pine pollen, 50-250 parts of schisandra chinensis, and an appropriate amount of starch. Alternatively, the Chinese medicine composition is prepared by adding the following Chinese medicine extracts in the following weight percentage ratios as the active ingredients, and adding excipients: 25-55% of Salvia miltiorrhiza extract, 12-40% of Gynostemma pentaphyllum extract, 5-12% of Schisandra chinensis alcohol extract, 12-30% of Cordyceps mycelium extract, 6-12% of pine pollen extract, 5-30% of peach kernel extract, and the rest are excipients; The above-mentioned pharmaceutical compound preparation wherein each effective component is prepared by the following method: ① Salvia miltiorrhiza extract Take Danshen medicinal material, crush it into granules, add about 12 times of water, extract twice at 95℃±2, combine the extracts and concentrate for later use. After testing, it is found that the water-soluble components of Danshen are mainly salvianolic acid B and sodium danshensu. Add about 70% ethanol to the residues and reflux twice, each time for about 1 hour, combine the extracts and concentrate for later use. After testing, the fat-soluble components of Danshen are mainly tanshinone. Combine the concentrated solutions and dry them for later use. ② Gynostemma pentaphyllum extract After the Gynostemma pentaphyllum medicinal material is soaked in about 75% ethanol, high-frequency ultrasonic extraction is performed three times to obtain the Gynostemma pentaphyllum extract, and the main component is total saponin after testing; ③Schisandra chinensis ethanol extract The raw material of Schisandra chinensis is added with about 80% ethanol for reflux extraction twice, and the extract is dried after the ethanol is recovered, and then crushed to obtain fine powder. After testing, the lignan components in Schisandra chinensis belong to the biphenylcyclooctene type lignan components, and Schisandrin A is the main component; the extraction rate of Schisandrin A by reflux extraction with about 80% ethanol is 0.3% to 2.0%; ④Cordyceps mycelium extract Take the Cordyceps mycelium raw material, add about 75% ethanol and warm extract three times, combine the extracts, recover the ethanol, and dry the extracts for later use; ⑤Pine pollen lipid extract Using about 70% ethanol to reflux heat to extract pine pollen, recovering ethanol from the extract, and drying the extract for later use; ⑥Peach kernel extract The effective components of the traditional Chinese medicine peach kernel are water-soluble components. The peach kernel is decocted twice with about 10 times the amount of water by water decoction method, and the extract is concentrated into an extract, and the extract is dried for later use. The above-mentioned extracts are dried by vacuum drying or spray drying. Wherein, the mass ratio between the compound of formula (I) and the traditional Chinese medicine composition is (1-150):(1-150).

2. The pharmaceutical composition according to claim 1, characterized in that The raw materials of the Chinese medicine composition include, by weight: 100-200 parts of peach kernel, 500-600 parts of salvia miltiorrhiza, 300-500 parts of gynostemma pentaphyllum, 200-300 parts of fermented cordyceps powder, 100-200 parts of pine pollen, 100-200 parts of schisandra chinensis, and an appropriate amount of starch; Preferably, the raw materials of the traditional Chinese medicine composition include, by weight: about 130 parts of peach kernel, about 530 parts of salvia miltiorrhiza, about 400 parts of gynostemma pentaphyllum, about 260 parts of fermented cordyceps powder, about 130 parts of pine pollen, about 130 parts of schisandra chinensis, and an appropriate amount of starch; More preferably, the mass ratio between the compound of formula (I) and the traditional Chinese medicine composition is (3-120):(8-120); Preferably, the mass ratio between the compound of formula (I) and the traditional Chinese medicine composition is (1-5):(5-10); Particularly preferably, the mass ratio between the compound of formula (I) and the traditional Chinese medicine composition is about 3: about 8; Particularly preferably, the mass ratio between the compound of formula (I) and the traditional Chinese medicine composition is (2.85-3.15):(7.6-8.4).

3. The pharmaceutical composition according to claim 1 or 2, characterized in that The pharmaceutical composition further comprises one or more drugs and / or extracts for preventing and treating diseases related to pulmonary fibrosis; Preferably, the pulmonary fibrosis is idiopathic pulmonary fibrosis; Preferably, the drug is selected from one or more of the following: tyrosine kinase inhibitors, antacids, antioxidants, anti-connective tissue growth factor antibodies, lysophosphatidic acid inhibitors, pentraxin-2, JAK-STAT signaling pathway inhibitors, phosphodiesterase 4 inhibitors and TAS-115; More preferably, the tyrosine kinase inhibitor is nintedanib; More preferably, the antacid is a proton pump inhibitor and / or a histamine 2 receptor antagonist; More preferably, the antioxidant is N-acetylcysteine.

4. The pharmaceutical composition according to claim 1 or 2, characterized in that The pharmaceutical composition further comprises one or more pharmaceutically acceptable excipients.

5. The pharmaceutical composition according to claim 4, characterized in that The auxiliary materials are selected from one or more of the following: diluents, wetting agents, binders, disintegrants, inclusion agents, flavoring agents, sustained-release agents, retention aids, lubricants, dispersants, plasticizers, opacifiers and antioxidants.

6. The pharmaceutical composition according to claim 4, characterized in that The dosage form of the pharmaceutical composition is powder, tablet, pill, capsule, film, lozenge, granule, injection or oral solution; More preferably, the dosage form of the pharmaceutical composition is a capsule or a tablet; Also preferably, the tablet is a dispersible tablet or a sustained-release tablet.

7. The pharmaceutical composition according to claim 1 or 2, characterized in that The preparation method of the Chinese medicine composition comprises the following steps: (1) adding water to decoct the salvia miltiorrhiza, peach kernel and gynostemma pentaphyllum weighed in weight ratio, combining the decoctions, taking the supernatant, concentrating it into an extract, cooling it, adding ethanol, precipitating it and filtering it; (2) weighing another portion of the fermented Cordyceps sinensis powder, adding ethanol to Schisandra chinensis and refluxing, cooling, combining the ethanol solution, filtering, and concentrating the filtrate to dryness; (3) weighing pine pollen and adding ethanol to warm immerse, combining the leaching solution, concentrating and drying to form a dry paste; and (4) Take the above three dry pastes, mix and grind them, add an appropriate amount of starch, mix well, and obtain the Chinese medicine composition.

8. Use of the pharmaceutical composition according to any one of claims 1 to 7 in the preparation of a medicament for preventing and treating diseases related to pulmonary fibrosis; Preferably, the pulmonary fibrosis is idiopathic pulmonary fibrosis; More preferably, the pharmaceutical composition is a drug that inhibits inflammation and fibrosis-related factors, regulates CD4 + The differentiation direction of T cell subsets plays a role in preventing and treating pulmonary fibrosis.

9. Use of the pharmaceutical composition according to any one of claims 1 to 7 in the preparation of a medicament for down-regulating the expression of cytokines selected from the group consisting of: IL-4, IL-6, IL-17, TNF-α, MMP-2, TIMP-1, CTGF, TGF-β1 and PDGF.

10. Use of the pharmaceutical composition according to any one of claims 1 to 7 in the preparation of a medicament for upregulating the expression of cytokines selected from the group consisting of: CD4 / CD8, IFN-γ, IL-2 and IL-10.

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