A preparation of Fagopyrum dibotrys solution for atomization inhalation, its preparation method and application

Abstract

The present invention belongs to the technical field of pharmaceutical preparations, and discloses a Fagopyrum dibotrys solution preparation for aerosol inhalation, its preparation method and application. The present invention provides a Fagopyrum dibotrys solution preparation for aerosol inhalation, including: the effective components of Fagopyrum dibotrys are obtained by extracting Fagopyrum dibotrys with an aqueous ethanol solution, subjecting it to alcohol precipitation, and then separating and purifying the obtained solution by resin adsorption. The present invention also provides a preparation method of the Fagopyrum dibotrys solution preparation for aerosol inhalation. The Fagopyrum dibotrys solution preparation for aerosol inhalation provided by the present invention is used in the application of drugs having the effects of enhancing the efficacy and reducing the toxicity in the radiotherapy and chemotherapy of lung tumors. This preparation is administered to patients by aerosol inhalation, directly delivering the drug to the lungs. It not only uses a small dose, has a quick onset of action, but also is safe, effective, has a high bioavailability, and has no other toxic side effects.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a Fagopyrum dibotrys solution preparation for aerosol inhalation, its preparation method and application. Background Art

[0002] Fagopyrum dibotrys is the dried rhizome of the plant Fagopryum dibtrys (D.DOn) Hara in the Polygonaceae family. It has the effects of clearing the lungs and expelling phlegm, clearing heat and detoxifying, expelling pus and detumescence, dispelling wind and removing dampness, etc. It also has cancer chemoprevention and anti-cancer activities, inhibits platelet aggregation, reduces reduction reactions and enhances macrophage ability.

[0003] The anti-tumor active ingredients of Fagopyrum dibotrys mainly exist in the rhizome part. A proanthocyanidin condensed tannin mixture with high anti-cancer activity is extracted from the root of Fagopyrum dibotrys, which is mainly composed of (-)-epicatechin and its dimer, called "Golden E". It can not only inhibit the growth of tumor cells but also enhance the immune function of the body. Due to the poor hygroscopicity and stability of "Golden E", when the dosage form of the "Golden E" medicament is a solid preparation, the capsule is relatively stable, but appropriate preservatives and stabilizers are often added during the preparation and storage processes, which cause certain damage to the human body. In addition, the drug loading capacity of the capsule is small, and the number of capsules taken orally by patients at one time is too large. Some patients experience stomach discomfort and digestive tract symptoms such as nausea after taking it, resulting in poor patient compliance, which greatly limits its clinical application.

[0004] With the increase in the number of clinical drug users, a single dosage form has been difficult to meet the clinical needs. As a new dosage form, the Fagopyrum dibotrys solution for aerosol inhalation has a faster onset and higher bioavailability compared with traditional dosage forms, and has broad clinical application prospects. Compared with conventional preparations, the advantages of inhalants are as follows: 1) Inhalants can directly enter the blood from the lungs and have a fast onset; 2) Local administration provides the accumulation of drugs in the target organ, increasing the curative effect while reducing toxic and side effects; 3) There is no first-pass effect and the bioavailability is high; 4) The aerosol inhalation preparation is a sterile preparation and no preservatives need to be added during the preparation process.

[0005] A solution preparation for aerosol inhalation with Chinese medicinal Fagopyrum dibotrys as the active ingredient disclosed in Patent CN109745364A is prepared by decocting Fagopyrum dibotrys with water, alcohol precipitation, and decolorization with activated carbon, and is used for the treatment of diseases such as acute lung abscess, acute and chronic tracheitis, asthmatic tracheitis, and bronchial asthma. However, there is currently no solution dosage form of Fagopyrum dibotrys for aerosol inhalation for the treatment of lung cancer diseases, which is difficult to meet the growing clinical needs. Summary of the Invention

[0006] In order to solve the above technical problems, the present invention provides a Fagopyrum dibotrys solution preparation for aerosol inhalation, improves the extraction method and preparation process of the active ingredients of Fagopyrum dibotrys, and the prepared Fagopyrum dibotrys solution preparation for aerosol inhalation is specifically used for the treatment of lung cancer diseases.

[0007] The technical solution of the present invention is as follows:

[0008] The present invention provides a Fagopyrum dibotrys solution preparation for atomization inhalation, and the solution preparation includes: effective components of Fagopyrum dibotrys, an osmotic pressure regulator, and a solvent, and a protective gas is introduced into the solution.

[0009] In the above-mentioned Fagopyrum dibotrys solution preparation for atomization inhalation, the effective components of Fagopyrum dibotrys are prepared by extracting Fagopyrum dibotrys with an ethanol aqueous solution, performing alcohol precipitation, and then subjecting the obtained solution to resin adsorption separation and purification.

[0010] As an implementation manner, the volume concentration of ethanol in the ethanol aqueous solution is 20%-60%, preferably 30%-40%; the extraction temperature is 20°C-70°C, preferably 30°C-60°C. The number of extractions is N times, where N is a natural number greater than or equal to 1, and N is preferably 1 or 2 or 3. After each extraction, filtration is performed to obtain N portions of filtrate, and the N portions of filtrate are combined.

[0011] As an implementation manner, before light-shielding cold storage and standing, alcohol precipitation, and resin adsorption separation, the solution is concentrated to improve the purification efficiency. The concentration is preferably carried out under reduced pressure at 50°C-60°C. The time for light-shielding cold storage and standing in the present invention is preferably 12-24 hours. During alcohol precipitation, the mass fraction of ethanol in the solution is 70%-90%. The stationary phase for the resin adsorption separation is macroporous resin, reverse-phase resin, or polyamide resin, and the mobile phase is an ethanol solution with a volume fraction of 30%-50%, preferably 35%-45% ethanol solution.

[0012] Among them, in each liter of the Fagopyrum dibotrys solution preparation for atomization inhalation, it includes the effective components of Fagopyrum dibotrys prepared from 100-2000 g of Fagopyrum dibotrys. As a preferred implementation manner, each liter of the Fagopyrum dibotrys solution preparation for atomization inhalation includes the effective components of Fagopyrum dibotrys prepared from 200-1800 g of Fagopyrum dibotrys, and further preferably the effective components of Fagopyrum dibotrys prepared from 250-1000 g of Fagopyrum dibotrys, such as 300 g, 500 g, 600 g, 900 g, 1200 g, 1600 g, 1650 g, 1700 g, 1800 g, or 1900 g, and the dosage ranges between any of these values. In the above-mentioned Fagopyrum dibotrys solution preparation for atomization inhalation, through reasonable adjustment of the concentration of the effective components of Fagopyrum dibotrys in the Fagopyrum dibotrys solution preparation for atomization inhalation, it is not only beneficial to its ability to form an aerosol, but also can improve its medicinal effect and reduce its irritation to the patient.

[0013] In the above-mentioned Fagopyrum dibotrys solution preparation for inhalation, the osmotic pressure regulator adjusts the osmotic pressure of the Fagopyrum dibotrys solution preparation to 280 - 320 mmol / L, more preferably 280 - 300 mmol / L. As a preferred embodiment, the osmotic pressure regulator is an inorganic salt or a saccharide. In each liter of the above-mentioned Fagopyrum dibotrys solution preparation for inhalation, as a preferred embodiment, the dosage of the inorganic salt osmotic pressure regulator is 1 - 15 g, such as 1.5 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, 10 g, 11 g, 12 g, 13 g or 14 g, and the dosage range between any of these values. The inorganic salt osmotic pressure regulator is preferably one or more of sodium chloride, magnesium chloride, and calcium chloride, more preferably sodium chloride; the saccharide osmotic pressure regulator is preferably one or more of glucose and fructose.

[0014] In the above-mentioned Fagopyrum dibotrys solution preparation for inhalation, as a preferred embodiment, the solvent is water or sterile injection water, preferably sterile injection water.

[0015] In the above-mentioned Fagopyrum dibotrys solution preparation for inhalation, as a preferred embodiment, it further includes a pH regulator. The pH regulator is selected from sodium hydroxide, potassium hydroxide, potassium dihydrogen phosphate, sodium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, sodium carbonate, and sodium bicarbonate, more preferably sodium hydroxide; as a preferred embodiment, the dosage of the pH regulator preferably makes the pH value of the Fagopyrum dibotrys solution preparation for inhalation be 3.5 - 7.5, more preferably 4.5 - 6.0; for example, the pH is 4.0, 4.5, 5.0, 5.5, 6.0, 6.5 or 7.0, and the pH value range between any of these values.

[0016] In the above-mentioned Fagopyrum dibotrys solution preparation for inhalation, the protective gas can be any one or several of carbon dioxide, nitrogen, and inert gas, preferably carbon dioxide. As a preferred embodiment, after introducing the protective gas, the residual oxygen content in the Fagopyrum dibotrys solution < 5 ppm, and the residual oxygen content in the headspace above the liquid surface < 2%, which can avoid the decomposition of the effective components of Fagopyrum dibotrys and improve its stability and storage time.

[0017] For the Fagopyrum dibotrys solution preparation for inhalation obtained by the present invention, an ultrasonic nebulizer, a compressed air nebulizer or a vibrating mesh nebulizer is used to inhale the Fagopyrum dibotrys solution for inhalation into the lungs in an atomized form, directly acting on the lesion site, with a fast onset and being safer and more effective.

[0018] The present invention also provides a preparation method of the above-mentioned Fagopyrum dibotrys solution preparation, including the following steps:

[0019] (1) Extract and concentrate Fagopyrum dibotrys with an ethanol aqueous solution to obtain Fagopyrum dibotrys concentrated solution 1;

[0020] (2) After the Fagopyrum dibotrys concentrated solution 1 is stored in the dark and refrigerated for 12 - 24 hours and then centrifuged, the supernatant is the Fagopyrum dibotrys solution 1;

[0021] (3) The Fagopyrum dibotrys solution 1 is concentrated to obtain a concentrated solution 2, subjected to alcohol precipitation, stored in the dark and refrigerated for 12 - 24 hours, and then filtered to obtain the Fagopyrum dibotrys solution 2;

[0022] (4) After removing alcohol from the Fagopyrum dibotrys solution 2, it is adsorbed, separated and purified by a resin column to obtain the active ingredient of Fagopyrum dibotrys;

[0023] (5) After the active ingredient of Fagopyrum dibotrys is stored in the dark and refrigerated for 24 - 48 hours, it is filtered, an osmotic pressure regulator and a pH regulator are added, filtered and filled, and a protective gas is introduced for sealing to obtain the Fagopyrum dibotrys solution for atomization inhalation.

[0024] As an embodiment, before the Fagopyrum dibotrys solution 2 is adsorbed, separated and purified by a resin column, it is pretreated first; preferably: the Fagopyrum dibotrys solution 2 is concentrated to remove alcohol to 1 / 2 - 2 times of the feeding amount.

[0025] As an embodiment, the filtration in step (5) is filtration with a 0.22 μm microporous membrane.

[0026] As an embodiment, a protective gas is introduced into the Fagopyrum dibotrys solution preparation until saturation. The introduction of carbon dioxide and / or nitrogen can be specifically any one of the following methods: 1) Dry ice is put into the filled Fagopyrum dibotrys solution, and the dry ice is converted into carbon dioxide to make the residual oxygen content of the Fagopyrum dibotrys solution < 5 ppm, and the residual oxygen content in the headspace above the liquid surface of the Fagopyrum dibotrys solution < 2%; 2) Carbon dioxide gas is directly filled under the liquid surface of the filled Fagopyrum dibotrys solution until the residual oxygen content of the Fagopyrum dibotrys solution < 5 ppm, and the residual oxygen content in the headspace above the liquid surface of the Fagopyrum dibotrys solution < 2%; 3) Nitrogen is directly introduced under the liquid surface of the filled Fagopyrum dibotrys solution until the residual oxygen content of the Fagopyrum dibotrys solution < 5 ppm, and the residual oxygen content in the headspace above the liquid surface of the Fagopyrum dibotrys solution < 2%.

[0027] The present invention also provides an application of the above-mentioned Fagopyrum dibotrys solution preparation for atomization inhalation in the preparation of a drug with synergistic and toxicity-reducing effects on radiotherapy and chemotherapy of lung tumors. Experiments prove that the Fagopyrum dibotrys solution for atomization inhalation of the present invention has an inhibitory effect on mouse Lewis lung cancer xenografts, and the tumor inhibition effect is better than that of oral Fagopyrum dibotrys capsules.

[0028] Compared with the prior art, the advantages of the present invention are as follows:

[0029] The solution preparation for nebulization inhalation of the present invention uses Fagopyrum dibotrys, a traditional Chinese medicine, as the raw material. It is extracted with an ethanol aqueous solution and purified by resin adsorption separation to obtain an extract with Fagopyrum dibotrys as the core labeled component. It is supplemented with an osmotic pressure regulator in the form of an aqueous solution at a specific concentration and has a certain pH value. It is introduced into a sterile, light-proof and sealed pharmaceutical system, and a protective gas is introduced to form a solution preparation for nebulization inhalation based on the active ingredients of Fagopyrum dibotrys. Adopting a combination of medicine and device, it is administered by nebulizer for the treatment of lung cancer patients. By adjusting the concentration of the active ingredients in Fagopyrum dibotrys and the synergistic effect of Fagopyrum dibotrys and the osmotic pressure regulator, the anti-cancer effect of the Fagopyrum dibotrys solution preparation for nebulization inhalation is enhanced. The Fagopyrum dibotrys solution preparation for nebulization inhalation of the present invention has excellent physical performance parameters and is easy to nebulize to form an aerosol; it also has the advantages of reliable quality, high safety, high stability and long storage time.

[0030] The present invention dissolves the active ingredients of Fagopyrum dibotrys in a suitable medium and delivers them to the lungs by oral inhalation in the form of an aerosol through a nebulizer. The drug is directly delivered to the site of action, reducing the distribution of the drug in other tissues and organs. Compared with oral administration, the total effective rate of the nebulization inhalation administration method is greatly improved, and it has the characteristics of safety, effectiveness, high bioavailability and low toxicity and side effects, so it has obvious advantages.

[0031] The Fagopyrum dibotrys solution preparation for anti-cancer prepared by the present invention fills the gap in the current market. The solution preparation prepared by the present invention is specially designed for patients for nebulization inhalation and is used in conjunction with a nebulizer, and the use process is convenient. Compared with Weimaining capsules, the administration route is different, the dosage is reduced, and the safety is improved. At the same time, it can avoid the damage caused to patients by the first-pass effect of the liver required for oral administration, increase the compliance of patients, and is a good administration route for the elderly and children who are inconvenient to take oral medicine.

[0032] The Fagopyrum dibotrys solution preparation for inhalation provided by the present invention is not only simple and easy to operate, has low production costs, but also is easy to industrialize. Description of the Drawings

[0033] Figure 1 Drug distribution after nebulization of the Fagopyrum dibotrys solution for nebulization inhalation by nebulizer A;

[0034] Figure 2 Drug distribution after nebulization of the Fagopyrum dibotrys solution for nebulization inhalation by nebulizer B;

[0035] Figure 3 Inhibitory effect of the Fagopyrum dibotrys solution for nebulization inhalation on Lewis lung cancer in mice. Detailed Embodiments

[0036] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without making creative efforts fall within the scope of protection of the present invention.

[0037] Example 1

[0038] Preparation of Fagopyrum dibotrys solution for atomization inhalation

[0039] (1) Take 300 g of Fagopyrum dibotrys, add 10 times 45% ethanol solution, extract at 40 - 50 °C for 3 times, each extraction time is 1 hour, filter separately, combine the filtrates, and concentrate and remove ethanol at 50 - 60 °C to 1 time the amount of Fagopyrum dibotrys per 1 mL to obtain Fagopyrum dibotrys concentrated solution 1;

[0040] (2) Let the Fagopyrum dibotrys concentrated solution 1 stand in the dark and refrigerate for 12 hours, centrifuge at 4000 r / min for 15 min, take the supernatant and concentrate it under reduced pressure at 50 - 60 °C to an appropriate amount, add absolute ethanol to make the ethanol content reach 90%, refrigerate and stand for 24 hours, filter, remove ethanol under reduced pressure at 50 - 60 °C, concentrate to 2 times the amount of Fagopyrum dibotrys per 1 mL, pass through a macroporous resin column, elute with 30% ethanol solution, collect the eluate, remove ethanol under reduced pressure at 50 - 60 °C, and add injection water to 1000 mL to obtain Fagopyrum dibotrys solution;

[0041] (3) Let the Fagopyrum dibotrys solution stand in the cold for 48 hours, filter, adjust the pH of the filtrate to 5.5 with sodium hydroxide solution, add 7.5 g of sodium chloride to adjust the osmotic pressure, filter through a 0.22 μm filter membrane and fill into containers, seal with carbon dioxide to obtain Fagopyrum dibotrys solution for atomization inhalation. The residual oxygen content of the solution is measured to be 2.1 ppm, and the residual oxygen content in the headspace is 0.9%.

[0042] Example 2

[0043] Preparation of Fagopyrum dibotrys solution for atomization inhalation

[0044] (1) Take 500 g of Fagopyrum dibotrys, add 10 times 35% ethanol solution, extract at 50 - 60 °C for 3 times, each extraction time is 1 hour, filter separately, combine the filtrates, and concentrate and remove ethanol at 50 - 60 °C to 1 time the amount of Fagopyrum dibotrys per 1 mL to obtain Fagopyrum dibotrys concentrated solution 1;

[0045] (2) Keep the Fagopyrum dibotrys concentrated solution 1 in the dark and refrigerate it for 12 hours, centrifuge at 4000 r / min for 15 min, take the supernatant, concentrate it under reduced pressure at 50 - 60 °C to an appropriate amount, add anhydrous ethanol until the alcohol content reaches 75%, refrigerate and stand for 24 hours, filter, remove the alcohol under reduced pressure at 50 - 60 °C, concentrate to 2 times the amount of Fagopyrum dibotrys used per 1 mL, pass through a SEP reverse resin column, elute with a 40% ethanol solution, collect the eluate, remove the alcohol under reduced pressure at 50 - 60 °C, add injection water to 1000 mL to obtain the Fagopyrum dibotrys solution;

[0046] (3) Keep the Fagopyrum dibotrys solution in the refrigerator and stand for 24 hours, filter, adjust the pH of the filtrate to 4.5 with sodium hydroxide solution, add 6 g of sodium chloride to adjust the osmotic pressure, filter through a 0.22 μm filter membrane and fill into containers, seal with carbon dioxide to obtain the Fagopyrum dibotrys solution for atomization inhalation. The residual oxygen content in the solution is measured to be 2.2 ppm, and the residual oxygen content in the headspace is 0.8%.

[0047] Example 3

[0048] Preparation of Fagopyrum dibotrys solution for atomization inhalation

[0049] (1) Take 1000 g of Fagopyrum dibotrys, add 10 times of 55% ethanol solution, extract at 50 - 60 °C for 3 times, each extraction time is 1 hour, filter separately each time, combine the filtrates, concentrate and remove the alcohol at 50 - 60 °C to 1 times the amount of Fagopyrum dibotrys used per 1 mL to obtain the Fagopyrum dibotrys concentrated solution 1;

[0050] (2) Keep the Fagopyrum dibotrys concentrated solution 1 in the dark and refrigerate it for 12 hours, centrifuge at 4000 r / min for 15 min, take the supernatant, concentrate it under reduced pressure at 50 - 60 °C to an appropriate amount, add anhydrous ethanol until the alcohol content reaches 85%, refrigerate and stand for 24 hours, filter, remove the alcohol under reduced pressure at 50 - 60 °C, concentrate to 2 times the amount of Fagopyrum dibotrys used per 1 mL, pass through a polyamide resin column, elute with a 45% ethanol solution, collect the eluate, remove the alcohol under reduced pressure at 50 - 60 °C, add injection water to 1000 mL to obtain the Fagopyrum dibotrys solution;

[0051] (3) Keep the Fagopyrum dibotrys solution in the refrigerator and stand for 24 hours, filter, adjust the pH of the filtrate to 5.5 with sodium hydroxide solution, add 4.5 g of sodium chloride to adjust the osmotic pressure, filter through a 0.22 μm filter membrane and fill into containers; add dry ice to the solution, seal after the dry ice is converted into carbon dioxide to obtain the Fagopyrum dibotrys solution for atomization inhalation. The residual oxygen content in the solution is measured to be 2.2 ppm, and the residual oxygen content in the headspace is 0.8%.

[0052] Example 4

[0053] Same as Example 2, except that after filling, nitrogen is introduced into the solution until saturated and then sealed. The residual oxygen content in the solution is measured to be 3.3 ppm, and the residual oxygen content in the headspace is 1.9%.

[0054] Examples 5 - 8

[0055] Same as Example 2, except that the pH of the filtrate was adjusted to 5, 5.5, 6 and 6.5 with 2% sodium hydroxide solution respectively.

[0056] Examples 9 - 12

[0057] Same as Example 4, except that the pH of the filtrate was adjusted to 5, 5.5, 6 and 6.5 with 2% sodium hydroxide solution respectively.

[0058] Comparative Example 1

[0059] Same as Example 2, except that 2 g of antioxidant sodium sulfite was added to the solution and carbon dioxide was not introduced into the solution.

[0060] Comparative Example 2

[0061] Same as Example 2, except that 2 g of antioxidant sodium metabisulfite was added to the solution and carbon dioxide was not introduced into the solution.

[0062] Comparative Example 3

[0063] Same as Example 2, except that 2 g of antioxidant sodium thiosulfate was added to the solution and carbon dioxide was not introduced into the solution.

[0064] Comparative Example 4

[0065] Same as Example 2, except that carbon dioxide was not introduced into the solution.

[0066] Comparative Example 5

[0067] Preparation of Fagopyrum dibotrys Capsules

[0068] (1) Take 500 g of Fagopyrum dibotrys, add 10 times of 35% ethanol solution, extract at 50 - 60 °C for 3 times, each extraction time is 1 hour, filter in batches, combine the filtrate, concentrate and remove ethanol at 50 - 60 °C to 1 time of the amount of Fagopyrum dibotrys per 1 mL to obtain Fagopyrum dibotrys concentrated solution 1;

[0069] (2) Let the Fagopyrum dibotrys concentrated solution 1 stand in the dark and refrigerate for 12 hours, centrifuge at 4000 r / min for 15 min, take the supernatant and concentrate it under reduced pressure to an appropriate amount at 50 - 60 °C, add anhydrous ethanol to make the alcohol content reach 75%, refrigerate and stand for 24 hours, filter, remove ethanol under reduced pressure at 50 - 60 °C, concentrate to 2 times the amount of Fagopyrum dibotrys per 1 mL, pass through a SEP reverse resin column, elute with 40% ethanol solution, collect the eluate, remove ethanol under reduced pressure at 50 - 60 °C, concentrate to a thick paste, vacuum dry at 60 °C, pulverize, sieve, add an appropriate amount of magnesium stearate, and fill into capsules (each capsule contains 0.4 g) to obtain.

[0070] Table 1: Specific implementation schemes of Examples 1 - 4 and Comparative Examples 1 - 4

[0071]

[0072] Table 2: Specific implementation schemes of Examples 5-12

[0073]

[0074]

[0075] Experimental Example 1

[0076] The content of the active ingredients in Examples 1-3 and Comparative Example 5 was detected by high performance liquid chromatography, and the results are shown as follows:

[0077] Table 3: Comparison of the content of active ingredients in Fagopyrum dibotrys solutions for aerosol inhalation at different concentrations

[0078]

[0079] Experimental Example 2

[0080] Determination of the stability of Fagopyrum dibotrys solution for aerosol inhalation

[0081] The samples prepared in Example 2, Example 4, Examples 5-12 and Comparative Examples 1-4 were placed in a stability test chamber at 40°C and a light box (4500 LX) for 30 days, and the color change of the solution, as well as the changes in pH value, procyanidin B 2 and epicatechin content were observed.

[0082] After being placed for 30 days under different protection conditions and in different environments, the color and state changes of the solution are shown in the following table:

[0083] Table 4: Color and state changes of Fagopyrum dibotrys solution for aerosol inhalation after being placed for 30 days under different conditions

[0084]

[0085] It can be seen from Table 4 that after the Fagopyrum dibotrys solution for aerosol inhalation is placed in a stability test chamber at 40°C and a light box (4500 LX) for 30 days under the protection of carbon dioxide or nitrogen, the color and state of the solution do not change. Under the conditions of no protection or adding 0.2% sodium sulfite, sodium metabisulfite, and sodium thiosulfate, the color of the solution quickly deepens, and there is even turbidity and precipitation, indicating poor stability.

[0086] After being placed for 30 days under different protection conditions and in different environments, the pH value changes of the Fagopyrum dibotrys solution are as follows:

[0087] Table 5: Changes in pH value of Fagopyrum dibotrys solution after being placed at 40°C for 30 days under different protection conditions

[0088]

[0089] Table 6: Changes in the pH value of the Fagopyrum dibotrys solution after being placed under light for 30 days under different protection conditions

[0090]

[0091] As can be seen from Table 5 and Table 6, for the Fagopyrum dibotrys solutions for nebulization inhalation with different pH values, after being placed in a 40 °C stability test chamber and a light box (4500 LX) for 30 days under the conditions of being protected by introducing carbon dioxide or nitrogen respectively, the pH of the solution did not change. However, under the conditions of no protection or adding 0.2% sodium sulfite, sodium metabisulfite, or sodium thiosulfate, the pH of the solution decreased significantly.

[0092] Under different protection conditions and in different environments, the changes in the content of procyanidin B in the Fagopyrum dibotrys solutions with different pH values after being placed for 30 days are as follows: 2 Content changes are as follows:

[0093] Table 7: Changes in the content of procyanidin B (mg / mL) in the Fagopyrum dibotrys solution after being placed at 40 °C for 30 days under different protection conditions 2 (mg / mL) Content changes

[0094]

[0095] Table 8: Changes in the content of procyanidin B (mg / mL) in the Fagopyrum dibotrys solution after being placed under light for 30 days under different protection conditions 2 (mg / mL) Content changes

[0096]

[0097]

[0098] As can be seen from Table 7 and Table 8, for the Fagopyrum dibotrys solutions for nebulization inhalation with different pH values, after being placed in a 40 °C stability test chamber and a light box (4500 LX) for 30 days under the protection of introducing carbon dioxide or nitrogen respectively, the content of procyanidin B in the solution did not change. Under the conditions of no protection or adding 0.2% sodium sulfite, sodium metabisulfite, or sodium thiosulfate, the content of procyanidin B 2 decreased significantly. 2 Content decreased significantly.

[0099] Under different protection conditions and in different environments, the changes in the content of epicatechin in the Fagopyrum dibotrys solutions with different pH values after being placed for 30 days are as follows:

[0100] Table 9: Changes in the content of epicatechin (mg / mL) in the Fagopyrum dibotrys solution after being placed at 40 °C for 30 days under different protection conditions

[0101]

[0102] Table 10: Changes in the content of epicatechin (mg / mL) in the Fagopyrum dibotrys solution under different protection conditions after being placed in the light for 30 days

[0103]

[0104]

[0105] As can be seen from Table 9 and Table 10, for the Fagopyrum dibotrys solution for nebulization inhalation with different pH values, after being placed in a 40°C stability test chamber and a light box (4500 LX) for 30 days, and under the protection of carbon dioxide or nitrogen being introduced respectively, the content of epicatechin in the solution did not change. Under the conditions of no protection, adding 0.2% sodium sulfite, sodium metabisulfite, and sodium thiosulfate, the content of epicatechin in the Fagopyrum dibotrys solution for nebulization inhalation decreased.

[0106] Experimental Example 3

[0107] Study on the atomization characteristics of the Fagopyrum dibotrys solution for nebulization inhalation

[0108] Two different nebulizers (commercial samples, divided into Type A and Type B) were selected, and the next-generation impactor (NGI) method was used to measure the aerodynamic particle size distribution. Parameters such as fine particle dose (FPD), mass median aerodynamic diameter (MMAD), and geometric standard deviation (GSD) were selected to characterize the in vitro atomization data of the Fagopyrum dibotrys solution for nebulization inhalation with procyanidin B 2

[0109] 1. Investigation of atomization parameters

[0110] Instrument parameters: The NGI instrument was selected for measurement, the flow rate was 15 L / min, and the operation was carried out after being placed in a cooling device at 5°C for 90 min. 5 mL of the Fagopyrum dibotrys solution for nebulization inhalation was accurately measured and placed in the atomization device for atomization. Type A and Type B nebulizers were used, both of which were compressed air nebulizers.

[0111] The atomization time of Type A and Type B nebulizers was investigated respectively. According to the different atomization parameters of Type A and Type B nebulizers, different atomization times were screened. Atomization was carried out for 1, 2, 3, 4, and 5 min respectively, and the NGI collection plates at all levels were visually inspected after atomization. It was observed that starting from 3 min, the atomized droplets on the collection plate showed obvious deformation and an overload phenomenon. Therefore, the atomization time was investigated for 1 and 2 min. The main indicators of the aerodynamic particle size distribution are fine particle dose (FPD), fine particle fraction (FPF), mass median aerodynamic diameter (MMAD), and geometric deviation (GSD), as shown in Table 11.

[0112] Table 11 Measurement results of atomization parameters of nebulizers of different brands (n = 3)

[0113] ​

[0114]

[0115] After atomization for 1 min and 2 min, the FPD increased with the prolongation of atomization time, but the FPF was basically the same, indicating that the proportion of fine particles has nothing to do with the length of atomization time. The atomization characteristic data such as MMAD and GSD showed little difference. However, due to the short atomization time of 1 min, the measurement error was large and the data variation was large. Therefore, after selecting atomization for 2 min, the aerodynamic particle size distribution of Fagopyrum dibotrys solution for inhalation aerosol was measured. The ideal GSD value of the aerosol is 1, but the aerosols of most drugs are non-uniform phases, and the GSD value is generally between 1.5 and 2.5. It can be seen from the above table that the Fagopyrum dibotrys solution for inhalation aerosol has excellent physical performance parameters, is easy to atomize to form an aerosol, and is beneficial to aerosol inhalation.

[0116] 2. Determination of aerodynamic particle size distribution

[0117] Using the screening parameters in the above experiment for determination, accurately measure 5 mL of Fagopyrum dibotrys solution for inhalation aerosol, and place it in the atomization device for atomization. Use Type A atomizer and Type B atomizer, and the atomization time is 2 min. NGI instrument, the flow rate is 15 L / min, and the flow rate variation range is (±5)%. After placing it in the cooling device at 5 °C for 90 min, perform the operation. The measurement results are shown in Figure 1 and Figure 2 .

[0118] From Figure 1 and Figure 2 , it can be seen that at a flow rate of 15 L / min, the resolution between NGI levels is relatively high, the particle sizes of the deposited particles between adjacent levels hardly overlap, the level loss is small, the data is reliable, and there is no need to use a separator.

[0119] Experimental Example 4

[0120] Inhibitory effect of Fagopyrum dibotrys solution for inhalation aerosol on transplanted Lewis lung tumor in mice

[0121] 1. Materials: Mouse Lewis lung cancer strain (Lewis lung carcinoma, 3LL).

[0122] 2. Animals: C57 BL / 6J mice, body weight (18 ± 2 g).

[0123] 3. Main reagents: Fagopyrum dibotrys solution for inhalation aerosol (Example 2), Fagopyrum dibotrys capsule (Comparative Example 5), cyclophosphamide CTX (commercially available).

[0124] 4. Model establishment: Lewis lung cancer-bearing mice were prepared. The tumor tissues were dissected, and well-growing masses were selected. They were homogenized at a ratio of tumor (g): normal saline (mL) of 1:3, and the cell count was adjusted to 2×10 6 / mL. Healthy C57 BL / 6J mice aged 6 - 8 weeks were selected, and 0.2 mL was subcutaneously inoculated into the right axilla of each mouse.

[0125] 5. Grouping and drug administration

[0126] After inoculation, the mice were randomly divided into 4 groups, with 15 mice in each group: normal saline group (NS), Fagopyrum dibotrys solution for aerosol inhalation group (Example 2 group, SSL2, 120 mg / kg), Fagopyrum dibotrys capsule group (Control Example 7 group, SSL19, 200 mg / kg), and cyclophosphamide group (CTX, 60 mg / kg). Drugs were administered on the second day after inoculation. For the Fagopyrum dibotrys solution for aerosol inhalation group, the mice were placed in a pet atomization box for aerosol inhalation administration; for the other groups, gavage administration was used for 15 consecutive days. The cyclophosphamide group was administered by intraperitoneal injection only once, and the mice were sacrificed by cervical dislocation on the 16th day.

[0127] 6. Tumor inhibition rate

[0128] The tumor tissues were weighed, and the tumor inhibition rate was calculated.

[0129] Tumor inhibition rate = (1 - average tumor weight of the experimental group / average tumor weight of the control group) × 100%

[0130] The inhibitory effect of Fagopyrum dibotrys solution for aerosol inhalation on Lewis lung cancer in mice (see Table 12, Figure 3 )

[0131] Table 12 Effects of Fagopyrum dibotrys solution for aerosol inhalation on tumor weight and tumor inhibition rate in mice (x±s, n = 15)

[0132]

[0133] From Table 12 and Figure 3 , it can be seen that the tumors in the saline control group were the heaviest; the tumors in the cyclophosphamide group were the lightest; the tumor weights in the Fagopyrum dibotrys solution for aerosol inhalation group were lighter than those in the Fagopyrum dibotrys capsule group; there were significant differences in the tumor weights compared with the control group (P < 0.01). The tumor inhibition rate in the Fagopyrum dibotrys solution for aerosol inhalation group was higher than that in the Fagopyrum dibotrys capsule group, reaching 49.61%.

[0134] The specific embodiments described above have further detailed the purpose, technical solutions, and beneficial effects of the present invention. It should be understood that the above description is only the specific embodiments of the present invention and is not used to limit the protection scope of the present invention. Any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A preparation of Fagopyrum dibotrys solution for atomization inhalation, characterized in that, the solution preparation includes: the active ingredient of Fagopyrum dibotrys, an osmotic pressure regulator and a solvent, and a protective gas is introduced into the solution; the active ingredient of Fagopyrum dibotrys is prepared by extracting Fagopyrum dibotrys with an ethanol aqueous solution, performing alcohol precipitation, and then subjecting the obtained solution to resin adsorption separation and purification; the volume concentration of ethanol in the ethanol aqueous solution is 20%-60%, and the extraction temperature is 20°C-70°C; the mobile phase for the resin adsorption separation and purification is an ethanol solution with a volume fraction of 30%-50%.

2. The Fagopyrum dibotrys solution preparation according to claim 1, characterized in that, the protective gas is an inert gas.

3. The Fagopyrum dibotrys solution preparation according to claim 1, characterized in that, the protective gas is any one or two of carbon dioxide and nitrogen.

4. The Fagopyrum dibotrys solution preparation according to claim 1, characterized in that, the stationary phase for the resin adsorption separation is macroporous resin, reversed-phase resin or polyamide resin.

5. The Fagopyrum dibotrys solution preparation according to claim 1, characterized in that, the residual oxygen content in the Fagopyrum dibotrys solution < 5 ppm, and the residual oxygen content in the headspace above the liquid surface < 2%.

6. The Fagopyrum dibotrys solution preparation according to claim 1, characterized in that, for each liter of the Fagopyrum dibotrys solution preparation for atomization inhalation, the dosage of Fagopyrum dibotrys is 100-2000 g, and the osmotic pressure of the Fagopyrum dibotrys solution preparation is 280-320 mmol / L.

7. The Fagopyrum dibotrys solution preparation according to claim 1, characterized in that, the pH value of the Fagopyrum dibotrys solution preparation for atomization inhalation is 3.5-7.

5.

8. The preparation method of the Fagopyrum dibotrys solution preparation according to any one of claims 1 to 7, characterized in that, it includes the following steps: (1) Extract and concentrate Fagopyrum dibotrys with an ethanol aqueous solution to obtain Fagopyrum dibotrys concentrated solution 1; (2) After the Fagopyrum dibotrys concentrated solution 1 is stored in the dark and refrigerated for 12-24 hours, centrifuge to obtain the supernatant as Fagopyrum dibotrys solution 1; (3) Concentrate the Fagopyrum dibotrys solution 1 to obtain concentrated solution 2, perform alcohol precipitation, store in the dark and refrigerate for 12-24 hours, and then filter to obtain Fagopyrum dibotrys solution 2; (4) After removing alcohol from the Fagopyrum dibotrys solution 2, perform resin column adsorption separation and purification to obtain the active ingredient of Fagopyrum dibotrys; (5) After the active ingredient of Fagopyrum dibotrys is stored in the dark and refrigerated for 24-48 hours, filter, add an osmotic pressure regulator and a pH regulator, filter and fill, and seal with a protective gas to obtain the Fagopyrum dibotrys solution preparation for atomization inhalation.

9. The preparation method of the Fagopyrum dibotrys solution preparation according to claim 8, characterized in that, when performing alcohol precipitation on the concentrated solution 2, the mass fraction of ethanol in the solution is 70%-90%.

10. The application of the Fagopyrum dibotrys solution preparation for atomization inhalation according to any one of claims 1 to 7 or the Fagopyrum dibotrys solution preparation for atomization inhalation prepared by the preparation method according to any one of claims 8 to 9 in the preparation of a drug for treating Lewis lung cancer.

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