Traditional Chinese medicine composition preparation for treating nephropathy as well as preparation method and quality control method thereof

Abstract

The invention discloses a traditional Chinese medicine composition preparation for treating nephropathy as well as a preparation method and a quality control method thereof, and belongs to the technical field of traditional Chinese medicine preparations. The traditional Chinese medicine composition preparation for treating nephropathy is prepared from the following components in percentage by weight: a traditional Chinese medicine composition and 1-90% of excipient, the traditional Chinese medicine composition is prepared from astragalus membranaceus, glossy privet fruit, radix paeoniae alba, bighead atractylodes rhizome, honeysuckle, curcuma zedoary and rhizoma sparganii. The preparation method is simple, granulation is smooth, particle forming is good, friability is low, the disintegration time of the prepared preparation is short, the defects that traditional Chinese medicine extract tablets are unstable in hygroscopicity and quality are overcome, and the quality control method is good in specificity, repeatability and stability and suitable for industrial production. The method can be used for quality control of traditional Chinese medicine composition preparations and traditional Chinese medicine compositions.

Description

Technical Field

[0001] The invention belongs to the technical field of traditional Chinese medicine pharmaceutical preparation, and particularly relates to a traditional Chinese medicine composition preparation for treating kidney disease, a preparation method thereof and a quality control method thereof. Background Art

[0002] Chronic glomerulonephritis (CGN), also known as chronic nephritis, is a primary glomerular disease characterized by multiple etiologies and pathological types, with chronic glomerular lesions being the predominant manifestation. Common clinical symptoms include hematuria, proteinuria, hypertension, and edema, most of which are insidious and develop slowly. Prolonged disease can progress to varying degrees of renal function impairment, renal fibrosis, and ultimately end-stage renal failure, threatening the lives of millions of patients. The disease is difficult to cure and prone to relapse, and modern medicine currently lacks a specific treatment.

[0003] The current treatment status of chronic glomerulonephritis is divided into the following categories: (1) General treatment: Patients with chronic nephritis who have edema, hypertension, large amounts of proteinuria, and renal function impairment should all get adequate rest and be given a high-quality low-protein diet according to their renal function status, while controlling the intake of phosphorus in food; (2) Control of hypertension: Effective control of hypertension plays an important role in protecting target organs and reducing target organ damage, especially controlling intrarenal capillary hypertension is an important measure to delay the progression of chronic renal failure; (3) Reduce proteinuria: Currently, the following types of drugs are mainly used to reduce urine protein in clinical practice: angiotensin-converting enzyme inhibitors (ACEI) and angiotensin II receptor antagonists (ARB), adrenal glucocorticoids, and immunosuppressants; (4) Improve renal microcirculation: The use of alprostadil can improve the patient's renal tissue blood rheology indicators by supplementing exogenous prostaglandin E1.

[0004] CN1565594A discloses a Chinese patent medicine for treating nephropathy and its preparation method. This Chinese patent medicine (i.e. Shenhua tablets) is made of Radix Astragali, Fructus Ligustri Lucidi, Radix Paeoniae Alba, Flos Lonicerae, Rhizoma Sparganii, Rhizoma Atractylodis Macrocephalae and Rhizoma Curcumae. Atractylodes Macrocephalae, Rhizoma Sparganii, Radix Paeoniae Alba and Flos Lonicerae are distilled with water to obtain volatile oil. Radix Astragali and Fructus Ligustri Lucidi are subjected to alcohol extraction to obtain an extractum. The residues after extracting the volatile oil and the residues after alcohol extraction are subjected to water extraction to obtain a clear paste. Finally, the extractum, the clear paste and the volatile oil are merged to make. Clinically, this Chinese patent medicine can reduce urine protein, serum creatinine and urinary nitrogen in patients with chronic nephritis and chronic renal insufficiency, improve plasma albumin, alleviate the pathological changes of the kidney, improve hemorheological indexes, and has a good therapeutic effect on kidney diseases such as chronic nephritis and chronic renal insufficiency.

[0005] How to further improve the efficacy of Chinese patent medicines in order to better treat kidney disease is an urgent problem that needs to be solved.

[0006] At the same time, there are currently problems in the preparation process of Shenhua tablets, such as difficulty in granulation and long disintegration time of the tablets during the dispersion process, and there is a lack of effective quality control methods. Therefore, it is urgent to standardize its production process and improve product quality and production controllability. Summary of the Invention

[0007] One of the purposes of the present invention is to provide a traditional Chinese medicine composition and its preparation, which can effectively treat kidney disease by adjusting the amount of raw materials and increasing the proportion of honeysuckle in the prescription.

[0008] The second purpose of the present invention is to provide a preparation method of the above-mentioned Chinese medicine composition preparation, standardize the production process, and provide technical support for industrial production.

[0009] The third object of the present invention is to provide a quality control method for the above-mentioned Chinese medicine composition preparation to improve the quality of the product.

[0010] In order to achieve the above object, the present invention adopts the following technical solutions:

[0011] In a first aspect, the present invention provides a Chinese medicine composition preparation, which is composed of a Chinese medicine composition and 1-90% of an excipient by weight.

[0012] The Chinese medicine composition comprises astragalus, glossy privet fruit, white peony root, atractylodes macrocephala, honeysuckle, zedoaria and trigonosciadium japonicum.

[0013] In some embodiments, the traditional Chinese medicine composition is made of the following raw materials in parts by weight: 61-119 parts of Astragalus, 61-119 parts of Ligustrum lucidum, 37-71 parts of White Peony, 37-71 parts of Atractylodes, 80-105 parts of Honeysuckle, 49-95 parts of Curcuma and 49-95 parts of Trigonosciadium japonicum.

[0014] In some embodiments, the traditional Chinese medicine composition is made of the following raw materials in parts by weight: 72-108 parts of Astragalus, 72-108 parts of Ligustrum lucidum, 42-66 parts of White Peony, 42-66 parts of Atractylodes, 80-105 parts of Honeysuckle, 54-90 parts of Curcuma and 54-90 parts of Trigonosciadium japonicum.

[0015] In some embodiments, the traditional Chinese medicine composition is made of the following raw materials in parts by weight: 88-92 parts of Astragalus, 88-92 parts of Ligustrum lucidum, 52-55 parts of White Peony, 52-55 parts of Atractylodes, 86-89 parts of Honeysuckle, 69-72 parts of Curcuma and 69-72 parts of Trigonosciadium japonicum.

[0016] In some embodiments, the extract of the traditional Chinese medicine composition is made from the following raw materials in parts by weight: 88-92 parts of Astragalus, 88-92 parts of Ligustrum lucidum, 52-55 parts of White Peony, 52-55 parts of Atractylodes, 87.8 parts of Honeysuckle, 69-72 parts of Curcuma and 69-72 parts of Trigonosciadium japonicum.

[0017] In one embodiment, the Chinese medicine composition is prepared from the following raw materials in parts by weight: 90.1 parts of Astragalus, 90.1 parts of Ligustrum lucidum, 53.5 parts of White Peony Root, 53.5 parts of Atractylodes Macrocephala, 87.8 parts of Honeysuckle, 70.9 parts of Curcuma and 70.9 parts of Trigonosciadium japonicum.

[0018] The astragalus described in the present invention includes astragalus medicinal materials and decoction pieces thereof. In some embodiments, the decoction pieces are processed astragalus;

[0019] The glossy privet fruit described in the present invention includes glossy privet fruit medicinal materials and decoction pieces thereof. In some embodiments, the decoction pieces are processed glossy privet fruit;

[0020] The white peony root described in the present invention includes white peony root medicinal materials and their decoction pieces. In some embodiments, the decoction pieces are processed white peony root;

[0021] The Atractylodes macrocephala described in the present invention includes Atractylodes macrocephala medicinal materials and their decoction pieces. In some embodiments, the decoction pieces are processed Atractylodes macrocephala. In some embodiments, the decoction pieces are bran-fried Atractylodes macrocephala.

[0022] The honeysuckle described in the present invention includes honeysuckle medicinal materials and decoction pieces thereof;

[0023] The zedoary turmeric described in the present invention includes zedoary turmeric medicinal materials and decoction pieces thereof. In some embodiments, the decoction pieces are processed zedoary turmeric;

[0024] The Trigonum gracile described in the present invention includes Trigonum gracile medicinal materials and their decoction pieces. In some embodiments, the decoction pieces are processed Trigonum gracile.

[0025] The astragalus, glossy privet fruit, white peony root, atractylodes macrocephala, honeysuckle, zedoaria and trillium described in the present invention are preferably prepared according to the processing method for the above-mentioned medicinal materials under the 2020 edition of the "Chinese Pharmacopoeia".

[0026] Through research, the inventors discovered that Astragalus membranaceus is sweet and slightly warm in nature, making it an excellent herb for tonifying the spleen and replenishing qi. The spleen is located in the middle of the body, like a dam in the middle, and therefore has the function of restraining the overflow of kidney water and consolidating the loss of kidney essence. Astragalus membranaceus is used to tonify the spleen by promoting earth and overcoming dampness, raising the clear and consolidating essence, thus achieving the effect of collecting earth and sealing the kidney. Ligustrum lucidum fruit is sweet, bitter, and cooling in nature, making it a key herb for nourishing the kidney and clearing away heat. When these two herbs are used together, Astragalus membranaceus replenishes qi and generates yin, while Ligustrum lucidum fruit nourishes yin and transforms qi, thus achieving a wonderful mutual transformation of qi and yin. Atractylodes macrocephala is bitter and sweet in nature, and can not only replenish qi and strengthen the spleen, but also dry dampness and invigorate the spleen, thus supporting Astragalus membranaceus's ability to promote earth and overcome dampness, consolidate the kidney and seal the meridians. White peony root is bitter, sour, and slightly cold in nature, and is good at nourishing blood and nourishing yin. When used with Ligustrum lucidum fruit, it can not only enhance the function of nourishing the liver and kidneys, but also have the effect of promoting blood circulation and promoting urination. Both Trigonosciadium japonicum and Curcuma zedoaria are pungent and warm in nature, and are effective in promoting blood circulation and removing blood stasis. Their combined efficacy is even stronger. Honeysuckle, on the other hand, is sweet and cold in nature, and is specialized in clearing heat and detoxifying. Based on the principles of this formula, the dosage of each component is adjusted to achieve optimal therapeutic efficacy in treating kidney disease.

[0027] In some embodiments, the method for preparing the traditional Chinese medicine composition comprises the following steps:

[0028] (1) Extracting volatile oil: extracting volatile oil from Atractylodes macrocephala and Curcuma zedoaria, and obtaining volatile oil, medicinal residue and extract I after solid-liquid separation;

[0029] (2) Alcohol extraction: Astragalus and Ligustrum lucidum are extracted with alcohol, and solid-liquid separation is performed to obtain alcohol extract and alcohol-extracted medicinal residue;

[0030] (3) Water extraction: extracting the medicinal residue obtained in step (1), the alcohol-extracted medicinal residue obtained in step (2), and the remaining raw materials by water extraction, and separating the solid and liquid to obtain aqueous extract II;

[0031] (4) Concentration: The extract I obtained in step (1) and the aqueous extract II obtained in step (3) are combined and concentrated to obtain aqueous extract concentrate III;

[0032] (5) alcohol precipitation: subjecting the aqueous extract concentrate obtained in step (4) to alcohol precipitation to obtain an alcohol precipitation supernatant;

[0033] (6) combining the alcohol extract obtained in step (2) and the alcohol precipitation supernatant obtained in step (5), and concentrating to obtain a thick paste;

[0034] (7) The thick paste obtained in step (6) is dried and mixed with volatile oil to obtain the product.

[0035] The extraction in step (1) of the present invention is a conventional volatile oil extraction process, such as distillation.

[0036] In some embodiments, the extraction in step (1) is a steam distillation method, including a heating distillation method after the raw material is immersed in water and a distillation method after the raw material is contacted with hot steam.

[0037] In one embodiment, the amount of water used in the extraction in step (1) is 1-8 times the total weight of Atractylodes macrocephala and Curcuma zedoaria, preferably 2-4 times, and most preferably 3 times.

[0038] In one embodiment, soaking is further included before the extraction; preferably, the soaking time is 0.2-8 hours, more preferably 0.2-6 hours, and most preferably 0.5 hours.

[0039] In one embodiment, the extraction time of the extraction in step (1) is 3-10 hours, preferably 4-6 hours, and most preferably 5 hours.

[0040] The alcohol extraction in step (2) of the present invention is a conventional process in the art, which utilizes the selectivity of alcohol solvents to dissolve and separate the active ingredients in traditional Chinese medicine.

[0041] In one embodiment, the alcohol extraction solvent used in the alcohol extraction in step (2) has an alcohol content of 50-80%, preferably 50-69%, and most preferably 60%.

[0042] In one embodiment, the amount of the alcohol extraction solvent used in the alcohol extraction in step (2) is 6-18 times the total weight of Astragalus and Ligustrum lucidum, preferably 6-12 times, and most preferably 10 times.

[0043] In one embodiment, the alcohol extraction in step (2) is performed 1-5 times, preferably 2-4 times, and most preferably 3 times.

[0044] In one embodiment, the extraction time of the alcohol extraction in step (2) is 1-5 hours, preferably 1-3 hours, and most preferably 1.5 hours.

[0045] In one embodiment, the alcohol extract in step (2) can be used directly without concentration or after concentration. In one embodiment, the concentration temperature is 50-75°C, preferably 50-70°C, and more preferably 60-70°C. In one embodiment, the relative density of the alcohol extract after concentration is 1.00-1.25 at 50-75°C, preferably 1.03-1.15, and more preferably 1.05-1.10.

[0046] The water extraction in step (3) of the present invention can be a conventional water extraction process in the art, such as room temperature extraction, hot water extraction, microwave-assisted extraction, steam distillation, ultrasonic-assisted extraction, aqueous phase extraction or countercurrent extraction.

[0047] In one embodiment, the amount of water used in the water extraction in step (3) is 6-18 times, preferably 6-14 times, and most preferably 10 times the total weight of the traditional Chinese medicine composition.

[0048] In one embodiment, the water extraction in step (3) is performed 1-4 times, preferably 1-3 times, and optimally 2 times.

[0049] In one embodiment, the water extraction time in step (3) is 1-5 hours, preferably 1-3 hours, and optimally 2 hours.

[0050] The concentration in step (4) of the present invention is performed by conventional methods in the art, such as freeze concentration, reduced pressure concentration or membrane concentration.

[0051] In one embodiment, the concentration in step (4) is concentration under reduced pressure.

[0052] In one embodiment, the concentration temperature in step (4) is 50-75°C, preferably 50-70°C, further 60-70°C.

[0053] In one embodiment, the relative density of the water-extracted concentrated solution III in step (4) is 1.10-1.40 at 50-75°C, preferably 1.10-1.30, and more preferably 1.15-1.20.

[0054] In one embodiment, the alcohol solvent used in the alcohol precipitation in step (5) is ethanol.

[0055] In one embodiment, after the alcohol solvent is added to the alcohol precipitation in step (5), the alcohol content is 50-80%, preferably 50-70%, and most preferably 60%.

[0056] In one embodiment, the temperature of the alcohol precipitation in step (5) is 0-10°C, preferably 2-8°C, more preferably 4-6°C, and most preferably 4°C.

[0057] In one embodiment, the standing time of the alcohol precipitation in step (5) is 12-84 hours, preferably 12-36 hours, and most preferably 24 hours.

[0058] In one embodiment, the alcohol precipitation supernatant in step (5) can be used directly without concentration or after concentration. In one embodiment, the concentration is concentrated under reduced pressure; in one embodiment, the concentration temperature is 50-75°C, preferably 50-70°C, and more preferably 60-70°C; in one embodiment, the relative density of the concentrated solution obtained by the concentration at 50-75°C is 1.00-1.25, preferably 1.05-1.15, and most preferably 1.05-1.10.

[0059] The concentration in step (6) of the present invention is concentration under reduced pressure.

[0060] In one embodiment, the concentration temperature in step (6) is higher than 45°C, preferably 45-90°C, more preferably 50-80°C, and even more preferably 55-75°C.

[0061] In one embodiment, the relative density of the thick paste in step (6) is 1.00-1.25 at 55-75°C, preferably 1.03-1.20, and more preferably 1.05-1.10.

[0062] In one embodiment, the relative density of the thick paste in step (7) is 1.15-1.45 at 55-75°C, preferably 1.20-1.40, and more preferably 1.25-1.30.

[0063] The drying in step (7) of the present invention is a conventional drying method in the art, including freeze drying, vacuum drying, oven drying or spray drying.

[0064] In one embodiment, the drying temperature in step (7) is 40-80°C, preferably 50-75°C, more preferably 50-70°C, and even more preferably 60-70°C.

[0065] The solid-liquid separation mentioned in the present invention is a conventional method in the art, such as centrifugation and / or filtration, which is not limited here.

[0066] In one embodiment, the preparation method comprises the following steps:

[0067] (1) Extracting volatile oil: Take Atractylodes macrocephala and Curcuma zedoaria, add 2-8 times water, soak for 0.2-8 hours, extract by steam distillation for 3-10 hours, collect volatile oil, filter the aqueous extract, and obtain aqueous extract I and medicinal residue for later use;

[0068] (2) Alcohol extraction: Take Astragalus and Ligustrum lucidum, add 6-18 times the amount of 50-80% ethanol, reflux and extract 1-5 times, each extraction time is 1-5 hours, filter, collect the alcohol solution, and concentrate under reduced pressure to obtain an alcohol extraction solution with a relative density of 1.00-1.25 (50-75°C) and alcohol extraction residue for use;

[0069] (3) Water extraction: Combine the medicinal residues and alcohol-extracted medicinal residues with the medicinal materials of Trigonella ternata, White Peony Root, and Honeysuckle, add 6-18 times the amount of water, reflux and extract 1-4 times, each extraction time is 1-5 hours, filter, and obtain aqueous extract II;

[0070] (4) Concentration: The aqueous extract I and the aqueous extract II are combined and concentrated to obtain an aqueous extract concentrate having a relative density of 1.10-1.40 (50-75° C.);

[0071] (5) Alcohol precipitation: Add ethanol to the aqueous extract concentrate to make the alcohol content reach 50-80%, let it stand at 0-10°C for 12-84 hours, filter to obtain the alcohol precipitate, and concentrate under reduced pressure to obtain the alcohol precipitation supernatant with a relative density of 1.00-1.25 (50-75°C);

[0072] (6) combining the alcohol extraction solution and the alcohol precipitation supernatant, and concentrating under reduced pressure to obtain a thick paste with a relative density of 1.15-1.45 (50-75°C);

[0073] (7) Dry the thick paste at 40-80°C and mix it with volatile oil to obtain the product.

[0074] In some embodiments, the Chinese medicine composition can also be prepared by subjecting the volatile oil obtained in step (1) of the above preparation method to inclusion treatment, and the inclusion compound used in the inclusion treatment is a conventional choice in the art, such as β-cyclodextrin.

[0075] In one embodiment, the preparation method of the traditional Chinese medicine composition is:

[0076] (1) Grind Atractylodes macrocephala and Curcuma zedoaria into coarse powder, add 3 times the amount of water, soak for 0.5 hours, extract for 5 hours, extract volatile oil, collect the aqueous solution and medicinal residue after extraction separately for later use; enclose the volatile oil in β-cyclodextrin, dry, and grind;

[0077] (2) Extracting Astragalus and Ligustrum lucidum by reflux extraction with 10 times the amount of 60% ethanol for 3 times, each time for 1.5 hours, and concentrating to obtain an alcohol extract with a relative density of 1.05-1.10. The alcohol-extracted medicinal residue was collected for later use;

[0078] (3) combining the alcohol-extracted medicinal residue obtained in step (2) and the medicinal residue obtained in step (1) with Trigonella ternate, white peony root, and honeysuckle, adding 10 times the amount of water and reflux extracting twice, each time for 2 hours, concentrating to obtain a water extract with a relative density of 1.18-1.20, precipitating with 60% ethanol, and letting the precipitate stand at 0-4°C for 24-48 hours. Taking the supernatant and concentrating it to obtain an alcohol precipitate with a relative density of 1.25-1.30;

[0079] (4) combining the alcohol extract obtained in step (2) and the alcohol precipitate obtained in step (3), drying them under vacuum or vacuum belt drying, pulverizing them, and mixing them with the inclusion compound obtained in step (1) to obtain the traditional Chinese medicine composition.

[0080] Preferably, the inclusion process in step (1) is as follows: grinding β-cyclodextrin with water, adding volatile oil ethanol solution dropwise after grinding evenly and continuing grinding, refrigerating, taking out and filtering, drying, and crushing to obtain volatile oil β-cyclodextrin inclusion complex.

[0081] Further preferably, the amount of β-cyclodextrin added is calculated by volume mass ratio (mL:g): volatile oil:β-cyclodextrin = 1:7-12; more preferably 10.

[0082] Further preferably, the amount of purified water added is calculated in terms of mass-to-volume ratio (g:mL): β-cyclodextrin: purified water = 1:2-6; further preferably, it is 4.

[0083] Further preferably, the volatile oil ethanol solution is prepared by dissolving the volatile oil in anhydrous ethanol.

[0084] More preferably, the grinding time is 20-60 minutes, and more preferably 30 minutes.

[0085] More preferably, the refrigeration is: refrigeration at -4-4°C for 12-24 hours; more preferably, refrigeration at 4°C for 24 hours.

[0086] More preferably, the drying is forced air drying, and the drying temperature is 35-45°C, and more preferably 40°C.

[0087] The excipients in the present invention include at least one of pharmaceutically acceptable fillers, disintegrants, lubricants and coating excipients.

[0088] In some embodiments, the filler is at least one of microcrystalline cellulose, dextrin and starch; preferably starch.

[0089] In some embodiments, the disintegrant is at least one of polyvinyl pyrrolidone, sodium carboxymethyl starch, starch, and low-substituted hydroxypropyl cellulose; preferably starch.

[0090] In some embodiments, the lubricant is at least one of talc, magnesium stearate and micronized silica gel; preferably magnesium stearate.

[0091] In some embodiments, the coating excipient is at least one of hydroxypropyl methylcellulose, Eudragit E100, and Opadry 290W665025-CN; preferably Opadry 290W665025-CN.

[0092] In some embodiments, the filler accounts for 15-25% of the weight of the traditional Chinese medicine composition, the disintegrant accounts for 1-4% of the weight of the traditional Chinese medicine composition, and the lubricant accounts for 0.1-0.5% of the weight of the traditional Chinese medicine composition.

[0093] In some embodiments, the weight percentage of the excipient in the traditional Chinese medicine composition preparation of the present invention is 5-85%, preferably 10-80%, more preferably 15-75%, and most preferably 20%.

[0094] In some embodiments, the filler and disintegrant account for 19.9% ​​of the weight of the traditional Chinese medicine composition, and the lubricant accounts for 0.1% of the weight of the traditional Chinese medicine composition.

[0095] In a second aspect, the present invention provides a method for preparing the above-mentioned Chinese medicine composition preparation, comprising the following steps: mixing the Chinese medicine composition with a filler and a disintegrant to form granules, adding a lubricant and tableting to obtain the Chinese medicine composition preparation.

[0096] The traditional Chinese medicine composition preparation prepared by the present invention has strong hygroscopicity. In order to make the preparation more stable, in some embodiments, the preparation method further includes a coating step after the tableting step.

[0097] In some embodiments, the coating process is as follows: the coating solid content is 15-20%, preferably 17%; the tablet bed temperature is 35-45°C, preferably 40°C; and the coating time is 50-60 min.

[0098] In a third aspect, the present invention provides the use of the above-mentioned Chinese medicine composition preparation and the Chinese medicine composition preparation prepared by the above-mentioned preparation method in the preparation of drugs for treating kidney disease.

[0099] The kidney disease described in the present invention includes chronic nephritis, proteinuria, or early or mid-stage renal insufficiency.

[0100] In one embodiment, the renal disease is chronic glomerulonephritis.

[0101] In one embodiment, the kidney disease is chronic nephritis with deficiency of both Qi and Yin, and blood stasis and toxins, or symptoms include fatigue, dry mouth and throat; dull complexion, hot palms and soles, soreness of waist and knees, fixed or stabbing pain in the waist, sore throat, pale red, reddish or dark red tongue, thin white or thin yellow or little coating, and thin, weak or thready pulse.

[0102] In some embodiments, the drug comprises the traditional Chinese medicine composition preparation (referred to as active ingredient) and pharmaceutically acceptable excipients.

[0103] The medicaments of the present invention may be prepared by any method known in pharmacy. Generally speaking, these preparation methods include associating the active ingredient with a carrier or excipient and / or one or more other auxiliary ingredients, and if necessary and / or desired, shaping and / or packaging the product into a desired single-dose or multi-dose unit.

[0104] The drug of the present invention can be prepared according to known methods, such as those described in the general rules for preparation of the Japanese Pharmacopoeia, 16th edition, the United States Pharmacopoeia, and the European Pharmacopoeia, 9th edition. The specific method depends on the dosage form.

[0105] The active ingredient and excipients in the medicaments of the present invention will vary depending on the identity, size and / or condition of the subject being treated and further on the route of administration of the medicament. The medicament may contain between 0.1% and 100% (w / w) active ingredient.

[0106] As used herein, "treating" unless otherwise indicated, means reversing, alleviating, inhibiting the progression of, or preventing the condition or disorder, or one or more symptoms of such condition or disorder, to which the term applies.

[0107] In a fourth aspect, the present invention provides a method for quality control of the above-mentioned Chinese medicine composition preparation, comprising the following steps:

[0108] S1. Preparation of test solution: dissolve the Chinese medicine composition preparation in water by ultrasonication, extract by shaking with water-saturated n-butanol, combine the n-butanol solutions, wash with ammonia test solution, discard the ammonia test solution, evaporate the n-butanol solution to dryness in a water bath, add water to the residue, dissolve it by ultrasonication, let it cool, pass it through a resin column, elute with water and ethanol respectively, discard the eluent, elute with ethanol again, collect the eluent, evaporate to dryness in a water bath, dissolve the residue in methanol, filter, and take the filtrate to obtain;

[0109] S2. Detection: Detect the content of astragaloside IV using high performance liquid chromatography.

[0110] In one embodiment, the resin in step S1 is a D101 macroporous resin.

[0111] In one embodiment, the volume percentage of ethanol in step S1 is 30-80%.

[0112] In one embodiment, the preparation method of the test solution in step S1 is: adding water to ultrasonically dissolve the Chinese medicine composition preparation, shaking and extracting with water-saturated n-butanol 4 times, 40 mL each time, combining the n-butanol solution, washing thoroughly 3 times with an ammonia test solution, 40 mL each time, discarding the ammonia test solution, and evaporating the n-butanol solution in a water bath. 5 mL of water is added to the residue, ultrasonically dissolved, cooled, passed through a D101 macroporous resin column (inner diameter 1.5 cm, column height 12 cm), eluted with 50 mL of water, discarded the water liquid, and then eluted with 30 mL of 40 v / v% ethanol, discarded the eluate, and then eluted with 80 mL of 70 v / v% ethanol, collected the eluate, evaporated to dryness in a water bath, dissolved in methanol, transferred to a 5 mL volumetric flask, added methanol to the scale, shaken, filtered, and the filtrate was obtained.

[0113] In one embodiment, the HPLC detection conditions in step S2 are: octadecylsilane bonded silica gel as filler; acetonitrile-water as mobile phase; evaporative light scattering detector detection; flow rate 0.8-1.2 mL / min; column temperature 25-35°C.

[0114] In one embodiment, the volume ratio of acetonitrile and water in step S2 is 30-40:60-70, preferably 31-33:67-69; further preferably 32:68; the drift tube temperature detected by the evaporative light scattering detector is 70-85°C, the atomization temperature is 70-80°C, and the gas flow rate is 1.2-2.0 L / min. Preferably, the drift tube temperature is 80°C, the atomization temperature is 75°C, and the gas flow rate is 1.6 L / min.

[0115] In one embodiment, the detection conditions of the high performance liquid chromatography in step S2 are: octadecylsilane bonded silica gel as the filler, acetonitrile-water (32:68) as the mobile phase; evaporative light scattering detector detection, drift tube temperature 80°C, nebulization temperature 75°C, gas flow rate 1.6 L / min; flow rate 1.0 mL / min; column temperature 30°C.

[0116] In one embodiment, step S1 further comprises the preparation of a reference solution: taking an astragaloside IV reference substance, adding methanol to prepare a solution.

[0117] In one embodiment, the reference solution is prepared by taking an appropriate amount of astragaloside IV reference substance, accurately weighing it, adding methanol to make a solution containing 0.5 mg per 1 mL, and shaking it well.

[0118] In one embodiment, step S1 further includes the preparation of a negative control solution: taking a negative extract that does not contain astragalus, and preparing it according to the preparation method of the test solution.

[0119] The Chinese medicine composition preparation described in the quality control method of the present invention can also be a Chinese medicine composition extract.

[0120] The beneficial effects of the present invention are:

[0121] (1) Based on the composition described in CN1565594A, the present invention adjusts the amount of raw materials and increases the proportion of honeysuckle in the prescription to prepare a traditional Chinese medicine composition preparation that can effectively treat kidney disease. Compared with the existing technology, the present invention has a better therapeutic effect on kidney disease.

[0122] (2) The preparation method of the Chinese medicine composition provided by the present invention is simple, the granulation is smooth, the granules are well formed, the friability is low, and the disintegration time is short.

[0123] (3) The Chinese medicine composition preparation provided by the present invention is coated with a moisture-proof coating liquid, which solves the shortcomings of hygroscopicity and unstable quality of Chinese medicine extract tablets and significantly reduces the moisture absorption rate compared with plain tablets.

[0124] (4) The quality control method of the present invention has good specificity, repeatability and stability, and can be used for the quality control of Chinese medicine composition preparations and Chinese medicine composition extracts. BRIEF DESCRIPTION OF THE DRAWINGS

[0125] Figure 1 This is the chromatogram of the reference substance in Example 13.1.

[0126] Figure 2 This is the chromatogram of the test sample in Example 13.1.

[0127] Figure 3 This is the negative control chromatogram in Example 13.1.

[0128] Figure 4 This is the linear relationship diagram of astragaloside IV in Example 13.3. DETAILED DESCRIPTION

[0129] The present invention is further described below with reference to the following examples, but these examples are not intended to limit the scope of the present invention.

[0130] The medicinal materials used in the following examples are as follows:

[0131] Astragalus is the dried root of Astragalus membranaceus (Fisch.) Bge.var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge. of the leguminous family.

[0132] Ligustrum lucidum fruit is the dried mature fruit of Ligustrum lucidum Ait., a plant of the Oleaceae family.

[0133] White peony root is the dried root of Paeonia lactiflora Pall., a plant of the Ranunculaceae family.

[0134] Atractylodes is the dried rhizome of Atractylodes macrocephala Koidz., a plant of the Asteraceae family.

[0135] Honeysuckle is the dried buds or initially opened flowers of Lonicera japonica Thunb. of the Caprifoliaceae family.

[0136] Curcuma zedoaria is the dried rhizome of Curcuma phaeocaulis Val., a plant of the ginger family.

[0137] Sparganium is the dried tuber of Sparganium stoloniferum Buch.-Ham., a plant of the Sparganaceae family.

[0138] Astragalus, Ligustrum lucidum fruit, white peony root, white atractylodes, honeysuckle, zedoaria, and trillium are prepared by drying fresh medicinal materials. According to the processing methods and preparation regulations of the above-mentioned medicinal materials in the 2020 edition of the "Chinese Pharmacopoeia", the various regulations were checked and the results showed that all the medicinal materials used in the test were in compliance with the pharmacopoeia regulations.

[0139] Example 1 Chinese medicine composition

[0140] Weigh 428 g of Astragalus, 428 g of Ligustrum lucidum fruit, 254 g of White Peony Root, 254 g of Atractylodes Macrocephala, 417 g of Honeysuckle, 337 g of Curcuma and 337 g of Sparganium as raw materials.

[0141] Extract volatile oil: Take Atractylodes macrocephala and Curcuma zedoariae and place them in a distiller, add 3 times the amount of water and soak for 0.5 hours, distill and extract for 5 hours, collect the volatile oil, filter the water extract, collect it in another container, and set the residue aside.

[0142] Alcohol extraction: Take Astragalus and Ligustrum lucidum, add 10 times the amount of 60% ethanol, reflux and extract 3 times, each extraction time 1.5 hours, filter, combine the filtrate, recover the ethanol, and concentrate under reduced pressure to an alcohol-extracted clear paste with a relative density of 1.07 (60°C). The residue is set aside.

[0143] Water extraction and concentration: the residues after alcohol extraction and volatile oil extraction are combined with the medicinal materials of Trigonum multiflorum, white peony root and honeysuckle, and reflux extraction is performed twice with 10 times the amount of water, each extraction is 2 hours, filtered, and the filtrate is combined with the distilled aqueous solution, and concentrated under reduced pressure to a water extract concentrate with a relative density of 1.15 (60°C).

[0144] Alcohol precipitation of water extract concentrate: add ethanol to the water extract concentrate to make the alcohol content reach 60%, refrigerate at 4℃ and let it stand for 24 hours. Take the supernatant and concentrate it under reduced pressure to an alcohol-precipitated clear paste with a relative density of 1.05 (60℃).

[0145] Concentration: Combine the alcohol precipitation paste and the alcohol extraction paste, and concentrate under reduced pressure to a thick paste with a relative density of 1.25 (60°C).

[0146] Combining: The thick paste is dried (60° C.), crushed into fine powder, and volatile oil is added for further combining and mixing to obtain a medicine for treating kidney disease.

[0147] Example 2 Chinese medicine composition

[0148] Weigh 445 g of Astragalus, 445 g of Ligustrum lucidum fruit, 277.5 g of White Peony Root, 277.5 g of Atractylodes macrocephala, 462.5 g of Honeysuckle, 360 g of Curcuma, and 360 g of Trigonosciadium japonicum as raw materials.

[0149] Extract volatile oil: Take Atractylodes macrocephala and Curcuma zedoariae and place them in a distiller, add 4 times the amount of water and soak for 0.2 hours, distill and extract for 6 hours, collect the volatile oil, filter the water extract, collect it in another container, and set aside the residue.

[0150] Alcohol extraction: Take the medicinal materials of Astragalus and Ligustrum lucidum, add 12 times the amount of 50% ethanol, reflux and extract 4 times, each extraction for 1 hour, filter, combine the filtrate, recover the ethanol, and concentrate under reduced pressure to an alcohol-extracted clear paste with a relative density of 1.05 (70°C), and the residue is set aside.

[0151] Water extraction and concentration: the residues after alcohol extraction and volatile oil extraction are combined with the medicinal materials of Trigonosciadium japonicum, White Peony Root, and Honeysuckle, and reflux extracted 3 times with 6 times the amount of water, each extraction time is 3 hours, filtered, and the filtrate is combined with the distilled aqueous solution, and concentrated under reduced pressure to a water extract concentrate with a relative density of 1.10 (70°C).

[0152] Alcohol precipitation of the water extract concentrate: add ethanol to the water extract concentrate to make the alcohol content reach 70%, refrigerate at 0℃ and let it stand for 48 hours. Take the supernatant and concentrate it under reduced pressure to an alcohol-precipitated paste with a relative density of 1.03 (70℃).

[0153] Concentration: Combine the alcohol precipitation paste and the alcohol extraction paste, and concentrate under reduced pressure to a thick paste with a relative density of 1.30 (70°C).

[0154] Combining: The thick paste is dried (40° C.), crushed into fine powder, and volatile oil is added for further combining and mixing to obtain a medicine for treating kidney disease.

[0155] Example 3 Chinese medicine composition

[0156] Weigh 595 g of Astragalus, 595 g of Ligustrum lucidum, 355 g of White Peony Root, 355 g of Atractylodes Macrocephala, 525 g of Honeysuckle, 475 g of Curcuma, and 475 g of Trigonosciadium japonicum as raw materials.

[0157] Extract volatile oil: Take Atractylodes macrocephala and Curcuma zedoariae and place them in a distiller, add 6 times the amount of water and soak for 2.5 hours, distill and extract for 8 hours, collect the volatile oil, filter the water extract, collect it in another container, and set aside the residue.

[0158] Alcohol extraction: Take the medicinal materials of Astragalus and Ligustrum lucidum, add 18 times the amount of 70% ethanol, reflux and extract once, each extraction time is 4 hours, filter, combine the filtrate, recover the ethanol, and concentrate under reduced pressure to an alcohol-extracted clear paste with a relative density of 1.10 (60°C), and the residue is set aside.

[0159] Water extraction and concentration: the residues after alcohol extraction and volatile oil extraction are combined with the medicinal materials of Trigonosciadium japonicum, White Peony Root, and Honeysuckle, and reflux extraction is performed twice with 10 times the amount of water, each extraction is 2 hours, filtered, and the filtrate is combined with the distilled aqueous solution, and concentrated under reduced pressure to a water extract concentrate with a relative density of 1.20 (60°C).

[0160] Alcohol precipitation of the water extract concentrate: add ethanol to the water extract concentrate to make the alcohol content reach 70%, refrigerate at 6℃ and let it stand for 12 hours. Take the supernatant and concentrate it under reduced pressure to an alcohol-precipitated paste with a relative density of 1.10 (60℃).

[0161] Concentration: Combine the alcohol precipitation paste and the alcohol extraction paste, and concentrate under reduced pressure to a thick paste with a relative density of 1.35 (55°C).

[0162] Combining: The thick paste is dried (50° C.), crushed into fine powder, and volatile oil is added for further combining and mixing to obtain a medicine for treating kidney disease.

[0163] Example 4 Chinese medicine composition

[0164] Weigh 540 g of Astragalus, 540 g of Ligustrum lucidum fruit, 360 g of White Peony Root, 360 g of Atractylodes Macrocephala, 900 g of Honeysuckle, 450 g of Curcuma, and 450 g of Trigonosciadium japonicum as raw materials.

[0165] Extract volatile oil: Take Atractylodes macrocephala and Curcuma zedoariae and place them in a distiller, add twice the amount of water and soak for 8 hours, distill and extract for 3 hours, collect the volatile oil, filter the water extract, collect it in another container, and set aside the residue.

[0166] Alcohol extraction: Take the medicinal materials of Astragalus and Ligustrum lucidum, add 18 times the amount of 80% ethanol, reflux and extract once, each extraction time is 2.5 hours, filter, combine the filtrate, recover the ethanol, and concentrate under reduced pressure to an alcohol-extracted clear paste with a relative density of 1.16 (75°C), and the residue is set aside.

[0167] Water extraction and concentration: Combine the residues after alcohol extraction and volatile oil extraction with the medicinal materials of Trigonum multiflorum, White Peony Root, and Honeysuckle, add 18 times the amount of water and reflux and extract once, each extraction time is 3.5 hours, filter, combine the filtrate with the distilled aqueous solution, and concentrate under reduced pressure to a water extract concentrate with a relative density of 1.25 (75°C).

[0168] Alcohol precipitation of the water extract concentrate: add ethanol to the water extract concentrate to make the alcohol content reach 75%, refrigerate at 8℃ and let it stand for 60 hours. Take the supernatant and concentrate it under reduced pressure to an alcohol-precipitated paste with a relative density of 1.15 (75℃).

[0169] Concentration: Combine the alcohol precipitation paste and the alcohol extraction paste, and concentrate under reduced pressure to a thick paste with a relative density of 1.40 (75°C).

[0170] Combining: The thick paste is dried (80° C.), crushed into fine powder, and volatile oil is added for further combining and mixing to obtain a medicine for treating kidney disease.

[0171] Example 5 Chinese medicine composition

[0172] Weigh 295 g of Astragalus, 295 g of Ligustrum lucidum fruit, 200 g of White Peony Root, 200 g of Atractylodes Macrocephala, 400 g of Honeysuckle, 245 g of Curcuma, and 245 g of Trigonosciadium japonicum as raw materials.

[0173] Extract volatile oil: Take Atractylodes macrocephala and Curcuma zedoariae and place them in a distiller, add twice the amount of water and soak for 1.5 hours, distill and extract for 4 hours, collect the volatile oil, filter the water extract, collect it in another container, and set aside the residue.

[0174] Alcohol extraction: Take the medicinal materials of Astragalus and Ligustrum lucidum, add 15 times the amount of 60% ethanol, reflux and extract 5 times, each extraction time is 3.5 hours, filter, combine the filtrate, recover the ethanol, and concentrate under reduced pressure to an alcohol-extracted clear paste with a relative density of 1.03 (65°C), and the residue is set aside.

[0175] Water extraction and concentration: the residues after alcohol extraction and volatile oil extraction are combined with the medicinal materials of Trigonum multiflorum, white peony root and honeysuckle, and reflux extraction is performed 3 times with 16 times the amount of water, each extraction is 5 hours, filtered, and the filtrate is combined with the distilled aqueous solution, and concentrated under reduced pressure to a water extract concentrate with a relative density of 1.30 (65°C).

[0176] Alcohol precipitation of water extract concentrate: add ethanol to the water extract concentrate to make the alcohol content reach 55%, refrigerate at 10℃ and let it stand for 36 hours. Take the supernatant and concentrate it under reduced pressure to an alcohol-precipitated paste with a relative density of 1.00 (65℃).

[0177] Concentration: Combine the alcohol precipitation paste and the alcohol extraction paste, and concentrate under reduced pressure to a thick paste with a relative density of 1.15 (80°C).

[0178] Combining: The thick paste is dried (70° C.), crushed into fine powder, and volatile oil is added for further combining and mixing to obtain a medicine for treating kidney disease.

[0179] Example 6 Chinese medicine composition

[0180] Weigh 360 g of Astragalus, 360 g of Ligustrum lucidum fruit, 180 g of White Peony Root, 180 g of Atractylodes Macrocephala, 360 g of Honeysuckle, 270 g of Curcuma, and 270 g of Trigonosciadium japonicum as raw materials.

[0181] Extract volatile oil: Take Atractylodes macrocephala and Curcuma zedoariae and place them in a distiller, add 8 times the amount of water and soak for 6 hours, distill and extract for 10 hours, collect the volatile oil, filter the water extract, collect it in another container, and set aside the residue.

[0182] Alcohol extraction: Take Astragalus and Ligustrum lucidum medicinal materials, add 6 times the amount of 80% ethanol, reflux extraction twice, each extraction time is 5 hours, filter, combine the filtrate, recover the ethanol, and concentrate under reduced pressure to an alcohol-extracted clear paste with a relative density of 1.20 (50°C), and the residue is set aside.

[0183] Water extraction and concentration: the residues after alcohol extraction and volatile oil extraction are combined with the medicinal materials of Trigonum multiflorum, white peony root and honeysuckle, and reflux extraction is performed 4 times with 14 times the amount of water, each extraction is 1 hour, filtered, and the filtrate is combined with the distilled aqueous solution, and concentrated under reduced pressure to a water extract concentrate with a relative density of 1.35 (50°C).

[0184] Alcohol precipitation of the water extract concentrate: add ethanol to the water extract concentrate to make the alcohol content reach 50%, refrigerate at 4°C and let it stand for 84 hours. Take the supernatant and concentrate it under reduced pressure to an alcohol-precipitated paste with a relative density of 1.20 (50°C).

[0185] Concentration: Combine the alcohol precipitation paste and the alcohol extraction paste, and concentrate under reduced pressure to a thick paste with a relative density of 1.20 (60°C).

[0186] Combining: The thick paste is dried (75° C.), crushed into fine powder, and further combined and mixed with volatile oil to obtain a medicine for treating kidney disease.

[0187] Example 7 Chinese medicine composition

[0188] Weigh 445 g of Astragalus, 445 g of Ligustrum lucidum fruit, 277.5 g of White Peony Root, 277.5 g of Atractylodes macrocephala, 462.5 g of Honeysuckle, 360 g of Curcuma, and 360 g of Trigonosciadium japonicum as raw materials.

[0189] Extract volatile oil: Take Atractylodes macrocephala and Curcuma zedoariae and place them in a distiller, add 4 times the amount of water and soak for 1 hour, distill and extract for 6 hours, collect the volatile oil, filter the water extract, collect it in another container, and set aside the residue.

[0190] Alcohol extraction: Take the medicinal materials of Astragalus and Ligustrum lucidum, add 12 times the amount of 50% ethanol, reflux and extract 4 times, each extraction for 1 hour, filter, combine the filtrate, recover the ethanol, and set aside the alcohol extract and the residue.

[0191] Water extraction and concentration: the residues after alcohol extraction and volatile oil extraction are combined with the medicinal materials of Trigonosciadium japonicum, White Peony Root, and Honeysuckle, and reflux extracted 3 times with 6 times the amount of water, each extraction time is 3 hours, filtered, and the filtrate is combined with the distilled aqueous solution, and concentrated under reduced pressure to a water extract concentrate with a relative density of 1.10 (70°C).

[0192] Alcohol precipitation of the aqueous extract concentrate: Add ethanol to the aqueous extract to make the alcohol content reach 70%, and refrigerate at 4°C for 48 hours. Take the supernatant for later use to obtain the alcohol precipitation supernatant.

[0193] Concentration: Combine the alcohol precipitation supernatant with the alcohol extract and concentrate under reduced pressure to a thick paste with a relative density of 1.25 (45°C).

[0194] Combining: The thick paste is dried (40° C.), crushed into fine powder, and volatile oil is added for further combining and mixing to obtain a medicine for treating kidney disease.

[0195] Example 8 Study on the efficacy of Chinese herbal medicine compositions prepared by different extraction processes on the SD rat adriamycin nephropathy model

[0196] The Chinese medicine composition of Example 1 was used as raw material to investigate the effects of extract powder A, extract powder B and extract powder C prepared according to processes A, B and C on the adriamycin nephropathy model of SD rats. Among them, (1) process A is water decoction, specifically: the Chinese medicine composition is decocted with water for 3 times, each time with 10 times the amount of water for 6 hours, the filtrate is combined, concentrated under reduced pressure, and dried to obtain Chinese medicine composition A; (2) process B adopts the extraction process of Example 1 in CN1565594A, specifically: Atractylodes macrocephala, Curcuma zedoaria, Trillium gracile, Paeonia lactiflora and Lonicera japonica are added with 10 times the amount of water, distilled for 8 hours to obtain volatile oil, and the medicinal liquid is stored in a separate container for standby use; Astragalus membranaceus and Ligustrum lucidum are refluxed and extracted twice with 80% ethanol, the first time with 5 times the amount and 2 hours, the second time with 5 times the amount and 4 hours, the ethanol solution is combined, the ethanol is recovered, and the extract is concentrated under reduced pressure to a relative density of 1.6. The residue after volatile oil extraction is combined with the residue after alcohol extraction, decocted with 6 times water for 4 hours, filtered, the filtered liquid and the liquid after volatile oil extraction are combined, and concentrated under reduced pressure to a clear paste with a relative density of 1.2, ethanol is added to make the alcohol content reach 60%, stirred, and allowed to stand for 30 hours. The supernatant is taken, ethanol is recovered under reduced pressure, concentrated to a clear paste with a relative density of 1.6, combined with the alcohol extract, and dried to obtain a dry powder; (3) Process C is the extraction process of Example 1 of the present invention.

[0197] 1. Experimental Animals

[0198] 180 male Sprague-Dawley rats, SPF grade. Weight at model establishment: 180-350 g. Animal use license number: SYXK (Su) 2016-0032. Room temperature and humidity control range: 20-26°C; 40-70%. Air changes: ≥15 / hour. Working lighting: ≥200 Lux; animal lighting: 15-20 Lux (lighting hours: 12 hours / day (lights on: 06:00 AM - off: 18:00 PM).

[0199] 2. Experimental animal grouping and dosage design

[0200] The proposed clinical dosage of extract powder C is 3.48g / day (calculated as extract powder). The average adult weight is 70kg, and the average SD rat weight is 300g. The equivalent dose for SD rats is 0.26g / kg. In this study, the doses of extract powder A, extract powder B, and extract powder C were all equivalent to 0.26g / kg. A normal control group, a model group, and a positive control group were also set up. The normal control group and the model group were gavaged with an equal volume of drinking water for the test animals, and the positive control group was given losartan potassium at a dose of 10mg / kg. A total of 6 groups were divided. Among them, the ratio of crude drug dosage corresponding to the same dose of extract powder B and extract powder C was 1.27:1.

[0201] The specific groups and numbers of animals are shown in Table 1 .

[0202] Table 1 Dosage groups for process screening pharmacodynamic studies

[0203]

[0204] Note: All groups were given the test substance, control substance or drinking water of the experimental animals at 1 mL / 100 g body weight.

[0205] 3. Test methods

[0206] Prepare doxorubicin 0.4 mg / mL and inject 1 mL / 100 g body weight of SD rats through the tail vein as a single injection. The modeling dose is 4 mg / kg. Detect 24-hour proteinuria 21 days after modeling. Screen rats with 24-hour urine protein greater than 44 mg and randomly group them according to the amount of 24-hour urine protein. Administer the drug on the day of grouping. Administer the drug for 8 weeks, and detect 24-hour urine protein at 4, 6, and 8 weeks of administration. Detect urine creatinine at the same time in the 8th week. The rats were killed the day after the last dose, and blood was collected from the abdominal aorta. The serum was centrifuged and the blood biochemical related indicators were measured. At the same time, the kidneys were taken and weighed, and the left kidney was taken for pathological examination.

[0207] 4. Administer medication

[0208] Administration: Oral administration.

[0209] Dosage volume: 1 mL / 100 g body weight.

[0210] Dosing cycle: 8 weeks.

[0211] 5. Detection indicators

[0212] 5.1 Weight

[0213] The experiment started with weekly weighing.

[0214] 5.2 General status observation

[0215] Observation time and frequency: After modeling or drug administration, observe the general condition of the animals every day;

[0216] Observation content: including but not limited to the animal's appearance, general behavior, posture, movement status, coat and skin, feces and urine, mental state, glandular secretion, respiratory status, fecal characteristics and survival period observation, etc.

[0217] 5.3 Blood collection and hematological indicators

[0218] Detection time: Detect after the end of drug administration.

[0219] Test animals: All living animals

[0220] Sampling method: Blood was collected from the abdominal aorta after the test.

[0221] Blood sample processing method: centrifuge at 3000g for 10 min and collect serum.

[0222] Test items and test methods: see Table 2 below.

[0223] Table 2 Blood biochemical test items

[0224]

[0225] 5.4 Urine testing

[0226] Testing time: once about two weeks after model establishment, and once about 4 weeks, 6 weeks, and 8 weeks after administration.

[0227] Test animals: All surviving animals.

[0228] Sampling method: 24-hour urine collection was performed using the metabolic cage method.

[0229] The test items and methods are shown in Table 3 below.

[0230] Table 3 Urine test items

[0231]

[0232] 5.5 Gross anatomy and histopathological examination

[0233] 5.5.1 Gross anatomy

[0234] Dissection time: After the experiment is completed.

[0235] Animals for autopsy: All living animals.

[0236] 5.5.2 Histopathological examination

[0237] Organ examination: Focus on observing changes in the kidney's texture, color, and volume. The left kidney of all surviving animals was sampled.

[0238] Animals examined: All animals.

[0239] Fixation method: Formalin fixation.

[0240] Preparation and staining.

[0241] 6. Statistical analysis

[0242] The results are presented in the form of charts, etc. SPSS23.0 software was used for statistics, and the data are expressed as mean ± standard deviation. The data were compared among the groups using one-way analysis of variance, and post hoc comparisons were performed. P < 0.05 indicated significant differences, and P < 0.01 indicated extremely significant differences.

[0243] 7. Test results

[0244] 7.1 Animal screening and grouping

[0245] A total of 170 animals were modeled in the experiment, and 24-hour urine protein was detected on the 21st day after modeling. The average urine protein in the normal control group was 29.2 mg. 55 rats with urine protein between 44.0-156.0 mg were screened and randomly divided into 5 groups according to the 24-hour urine protein range, namely model group, extract powder A, extract powder B, extract powder C and positive control group.

[0246] 7.2 Animal Death and General Condition Observation

[0247] All animals in the experiment had smooth fur, normal behavior, and normal feces and urine. No animals died during the experiment.

[0248] 7.3 weight

[0249] The body weight of each group during the experiment is shown in Table 4.

[0250] Table 4 Body weight results of each group

[0251]

[0252] Note: Compared with the normal control group, *p<0.05, **p<0.01.

[0253] The results showed that compared with the normal control group, the body weights of the animals in the other groups showed extremely significant differences (p < 0.01) after four weeks of administration, and extract powder C had a better effect on body weight in all treatment groups.

[0254] 7.4 24-hour urine protein results

[0255] The 24-hour urine protein test results of each group of animals during the experiment are shown in Table 5.

[0256] Table 5 24hr urine protein results

[0257]

[0258] Note: Compared with the normal control group, *p<0.05, **p<0.01; compared with the model group, #p<0.05, ##p<0.01.

[0259] The results showed that before administration, compared with the normal control group, the 24-hour urine protein of rats in other groups was extremely significantly increased (p < 0.01), indicating that the model was successful. Compared with the model group, there was no significant difference in the 24-hour urine protein of rats in the extract powder A, B, C and positive control groups (p > 0.05), and the groups were evenly divided.

[0260] After six weeks of administration, the extract powder C group showed a significant decrease in 24-hour urine protein compared to the model group (p < 0.05). This experiment demonstrates that the Chinese herbal composition prepared using the extraction process of the present invention is fast-acting and superior to the Chinese herbal compositions prepared using processes A and B in reducing 24-hour urine protein, and also superior to positive-acting drugs.

[0261] 7.5 Kidney weight and renal index

[0262] The results of the left and right kidneys and renal indexes of the animals in each group after 8 weeks of drug administration are shown in Table 6.

[0263] Table 6 Kidney weight and renal index results

[0264]

[0265] Note: Compared with the normal control group, *p<0.05, **p<0.01.

[0266] The results showed that after 8 weeks of administration, compared with the normal control group, except for the extremely significant increase in the renal index of rats in the model group (p < 0.01), there was no significant difference in the renal index of rats in other groups (p > 0.05).

[0267] 7.6 Blood biochemistry test results

[0268] The blood biochemical results of animals in each group after 8 weeks of drug administration are shown in Table 7.

[0269] Table 7 Animal blood biochemical results

[0270]

[0271] Note: Compared with the normal control group, *p<0.05, **p<0.01.

[0272] The results showed that compared with the normal control group, the serum ALB level of rats in the model group was extremely significantly reduced (p < 0.01). However, compared with the model group, the ALB levels of extract powder B, extract powder C, and the positive control group increased, but there was no significant difference compared with the normal group. The results showed that, compared with the model group, extract powders B, C, and the positive control group, with the exception of the traditional Chinese medicine composition prepared by process A, all had the effect of increasing ALB levels in rat serum.

[0273] 7.7 Pathological test results

[0274] The left kidney was routinely stained with HE and examined under a light microscope. Scoring was based on the severity of basophilic tubules, proteinaceous casts, glomerular degeneration, and interstitial inflammatory cell infiltration. No lesions were scored as 0, and 1, 2, 3, and 4 indicated mild, moderate, and severe lesions, respectively.

[0275] Pathological results showed that compared with the normal control group, the model group showed pathological changes such as basophilic renal tubules, tubular protein casts, swelling and degeneration of glomerular visceral epithelial cells, and interstitial inflammatory cell infiltration in the kidneys. Compared with the model group, each treatment group showed similar pathological changes. The degree of lesions in the extract powder group A was not significantly reduced compared with the model group, while the degree of lesions in the extract powder group B was slightly reduced compared with the model group. The degree of renal lesions in the extract powder group C and the positive control group was significantly reduced compared with the model group. This shows that compared with conventional processes, the Chinese medicine composition prepared by the extraction process of the present invention is more effective in treating kidney disease.

[0276] Example 9 Preparation of Chinese medicine composition

[0277] Weigh 254g of Atractylodes macrocephala and 337g of Curcuma zedoaria respectively, grind them into coarse powder, add 3 times the amount of water, soak for 0.5 hours, steam distill and extract for 5 hours, collect the volatile oil, and collect the aqueous solution and the medicinal residue after extraction for use; the volatile oil is included in the β-cyclodextrin (take 10 times the amount of β-cyclodextrin of the volatile oil, add 4 times the amount of purified water of β-cyclodextrin, grind it thoroughly, grind it evenly, slowly add the volatile oil ethanol solution dropwise, grind it for 30 minutes, refrigerate it at 4°C for 24h, take it out, filter it, and dry it with air at 40°C), grind it to obtain the volatile oil β-cyclodextrin inclusion complex; weigh 428g of Astragalus membranaceus and 428g of Ligustrum lucidum respectively, add 10 times the amount of 60v / v% The method comprises the following steps: extracting with ethanol under reflux for 3 times, each time for 1.5 hours, and concentrating to obtain an alcohol extract with a relative density of 1.05-1.10; combining the alcohol-extracted medicinal residues and the medicinal residues after volatile oil extraction with 337 g of Trigonum multiflorum, 254 g of white peony root, and 417 g of honeysuckle, adding 10 times the amount of water and extracting under reflux for 2 times, each time for 2 hours, and concentrating to obtain a water extract with a relative density of 1.18-1.20; precipitating with 60 v / v% ethanol, and allowing the precipitate to stand at 0-4° C. for 24-48 hours, collecting the supernatant and concentrating to obtain an alcohol precipitate with a relative density of 1.25-1.30, combining the precipitate with the alcohol extract, vacuum drying, crushing, and mixing with a volatile oil β-cyclodextrin inclusion compound to obtain a traditional Chinese medicine composition.

[0278] The moisture content of this product is controlled within 8%.

[0279] Example 10 Study on the Forming Process of Chinese Medicine Composition Preparation

[0280] Determination of disintegration time: According to the provisions of the 2015 edition of the Chinese Pharmacopoeia, Part IV (General Rule 0101) Tablets, and according to the disintegration time test method (General Rule 0921), take 6 plain tablets of this product, and all of them should disintegrate within 60 minutes.

[0281] Example 10.1 Selection of fillers

[0282] Dextrin, starch, and microcrystalline cellulose were selected as the main excipients for granule forming. The granule forming condition, tablet disintegration time, and hardness were used as the evaluation indicators to conduct a comparative study on the filling types. The specific experiments are as follows:

[0283] Take the traditional Chinese medicine composition prepared by the preparation method of Example 9, weigh 20 g per portion, weigh 3 portions in total, weigh 3.7 g each of dextrin, starch, and microcrystalline cellulose, add 1.3 g of sodium carboxymethyl starch (CMS-Na) to each group, mix well, and the preparation molding formula is shown in Table 8.

[0284] Table 8 Formulation Formula

[0285]

[0286] According to the above formulation, granules were produced using a dry granulator (feed power: 10 Hz; roller power: 20 Hz; granulation power: 10 Hz; pressure: 2 MPa). The granulation process was observed to examine the effects of the various excipients on granulation. The granules were sieved through a 65-mesh sieve. 0.1% magnesium stearate was added to each granule, mixed, and then tableted. Tablet hardness and disintegration time were used as evaluation criteria. The results are shown in Table 9.

[0287] Table 9 Effects of different fillers on granule forming and tablets

[0288]

[0289] The experimental results show that, based solely on formability, the use of dextrin, starch, and microcrystalline cellulose can all meet the needs of formulation forming, and the disintegration time is all within 15 minutes. Among them, the tablets using starch as a filler disintegrate the fastest. The disintegration effects of the three tablets are: starch > dextrin > microcrystalline cellulose, so starch is selected as the filler.

[0290] Example 10.2 Investigation of filler dosage

[0291] The granulation conditions, tablet disintegration time, and hardness of the tablets with a weight ratio of the Chinese medicine extract to starch of 3:1, 4:1, and 5:1 were compared. The specific experiments are as follows:

[0292] The Chinese medicine composition prepared according to the preparation method of Example 9 was taken, and 20 g of each portion was weighed. Three portions were weighed, and 5.7 g, 3.7 g, and 2.7 g of starch were added, respectively. 1.3 g of sodium carboxymethyl starch (CMS-Na) was added to each group. The mixture was uniformly mixed and dry granulated (feeding power: 10 Hz; roller power: 20 Hz; granulation power: 10 Hz; pressure: 2 MPa). The granulation phenomenon was observed and the effect of the different excipient ratios on granulation formation was investigated. The granules were sieved through a 65-mesh sieve. 0.1% magnesium stearate was added to the resulting granules, mixed, and tableted. The hardness and disintegration time of the plain tablets were used as the evaluation indicators. The results are shown in Table 10.

[0293] Table 10 Filler dosage investigation

[0294]

[0295] The experimental results show that when the ratio of extract powder to starch is 5:1, granulation is difficult and it is difficult to granulate normally. When the ratio is between 3:1-4:1, the granulation process is smooth, the granule formability is good, and the hardness and disintegration time meet the requirements. Considering factors such as tablet weight, the ratio of dry extract powder to starch is selected as 4:1, that is, the starch dosage is 20%.

[0296] Example 10.3 Investigation of disintegrant types and dosages

[0297] Considering that this product is easy to absorb moisture, a coating process is required. To ensure the normal disintegration of the tablets after coating, the effects of the type and dosage of disintegrants on the tablets were investigated during the preparation process. Cross-linked polyvinyl pyrrolidone PVPP, low-substituted hydroxypropyl cellulose L-HPC, and sodium carboxymethyl starch CMS-Na were selected as disintegrants. The specific experiments are as follows:

[0298] The Chinese medicine composition prepared according to the preparation method of Example 9 was weighed, and 20 g of each portion was weighed for a total of 3 portions. 4 g of starch was added to each portion. 1 g each of cross-linked polyvinyl pyrrolidone (PVPP), low-substituted hydroxypropyl cellulose (L-HPC), and sodium carboxymethyl starch (CMS-Na) was added to each group. The mixture was uniformly mixed and dry granulated (feeding power: 10 Hz; roller power: 20 Hz; granulation power: 10 Hz; pressure: 2 MPa). The granules were sieved through a 65-mesh sieve. 0.1% magnesium stearate was added to the resulting granules, mixed, and tableted. The hardness and disintegration time of the plain tablets were evaluated. The results are shown in Table 11.

[0299] Table 11 Investigation of disintegrant types

[0300]

[0301] The experimental results show that PVPP has the lowest hardness and the shortest disintegration time when used as a disintegrant, so PVPP is tentatively selected as the disintegrant.

[0302] Weigh 20g of the traditional Chinese medicine composition prepared according to the method of Example 9, for a total of four portions. PVPP was added at 0%, 1%, 2%, and 4% concentrations, respectively. An appropriate amount of starch was added to achieve a 4:1 ratio of extract powder to excipients. Mix thoroughly, dry granulate (feeding power: 10 Hz; roller power: 20 Hz; granulation power: 10 Hz; pressure: 2 MPa), and sieve through a 65-mesh sieve. The resulting granules were then added with 0.1% magnesium stearate, mixed thoroughly, and tableted. Tablet hardness and disintegration time were used as evaluation criteria. The results are shown in Table 12.

[0303] Table 12 Investigation of disintegrant dosage

[0304]

[0305] The results show that when the disintegrant PVPP dosage is 0-4%, the material fluidity, dry granulation and tableting process formability are all good, and the addition of disintegrant has no significant difference in the disintegration time of the plain tablets.

[0306] Taking into account the fact that the types and dosages of various disintegrants did not significantly shorten the disintegration time of the plain tablets, and that the plain tablets were completely disintegrated within 15 minutes when starch was used as a filler, starch was finally selected as the formulation filler and disintegrant.

[0307] Example 10.4 Investigation of lubricant dosage

[0308] The Chinese medicine composition prepared according to the preparation method of Example 9 was taken, and 20 g of each portion was weighed. A total of 3 portions were weighed, and 5 g of starch was added to each portion. The mixture was uniformly mixed and granulated using a dry granulator (feed power: 10 Hz; roller power: 20 Hz; granulation power: 10 Hz; pressure: 2 MPa). The granules were sieved through a 65-mesh sieve. 0.1%, 0.3%, and 0.5% of magnesium stearate were added to each granule, respectively. The mixture was uniformly mixed and tableted. The lubricant was evaluated using tablet hardness and disintegration time as evaluation indicators. The results are shown in Table 13.

[0309] Table 13 Investigation of lubricant dosage

[0310]

[0311] The results show that when the dosage of magnesium stearate is 0.1-0.5%, the tablet hardness and disintegration time are better, and 0.1% magnesium stearate is proposed to be used as a lubricant.

[0312] Example 11 Study on the coating process of Chinese medicine composition preparation

[0313] Considering that this product is easy to absorb moisture and stick together, it is necessary to conduct coating process research. The specific experiments are as follows:

[0314] 100 tablets prepared in Example 10.4 with a 0.1% lubricant were taken. Three replicates were coated with Opadry 290W665025-CN as a coating excipient. Coating solutions with solids contents of 16%, 17%, and 18% were prepared, respectively. The tablet bed temperature was 40°C. Conventional coating techniques were used, with the inlet air temperature controlled at 85-105°C, the outlet air temperature at 50-70°C, and the peristaltic pump speed at 50-100 rpm. The tablet weight gain was 3%-5%. The properties of the film-coated tablets were observed during the coating process. The results showed that all three coating solution concentrations were suitable for coating. Considering the difficulty of preparing the coating solution and the drying time of the coated tablets, a coating solution with a solids content of 17% was used as the coating solution.

[0315] Example 12 Preparation of Chinese medicine composition preparation

[0316] Based on the above research, 100 g of the traditional Chinese medicine composition prepared according to the method of Example 9 was added with 25 g of starch, mixed, dry granulated, sieved through a 65-mesh sieve, and 0.1% magnesium stearate was added. The mixture was mixed, compressed, and film-coated using Opadry 290W665025-CN as a coating excipient with a coating liquid solid content of 17%. The tablets were packaged to obtain the product. The results of the preparation process evaluation indicators are shown in Tables 14 and 15.

[0317] Table 14 Test results of the preparation process of Chinese medicine composition

[0318]

[0319] Table 15 Test results of moisture absorption performance of Chinese medicine composition preparations under different humidity conditions

[0320]

[0321] Compared with plain tablets, the hygroscopicity of the film-coated tablets prepared by the process provided by the present invention is significantly reduced under different humidity conditions.

[0322] Example 13 Quality Control of Chinese Medicine Composition

[0323] 1. Chromatographic conditions and system suitability test

[0324] Octadecylsilane-bonded silica gel was used as the filler; acetonitrile-water (32:68) was used as the mobile phase; detection was performed using an evaporative light scattering detector (drift tube temperature 80°C, nebulizer temperature 75°C, gas flow rate 1.6 L / min); flow rate 1.0 mL / min; column temperature 30°C. The theoretical plate number, calculated based on the astragaloside IV peak, should be no less than 4000.

[0325] 2. Preparation of reference solution

[0326] Take an appropriate amount of astragaloside IV reference substance, weigh accurately, add methanol to make a solution containing 0.5 mg per 1 mL, shake well, and obtain.

[0327] 3. Preparation of test solution

[0328] Accurately weigh 2.0 g of dry extract powder (prepared according to the preparation method of Example 9, batch number: 2018091003), add 10 mL of water, dissolve it by ultrasound, extract it with n-butanol saturated with water by shaking 4 times, 40 mL each time, combine the n-butanol solution, wash it thoroughly with ammonia test solution 3 times, 40 mL each time, discard the ammonia test solution, evaporate the n-butanol solution in a water bath, add 5 mL of water to the residue, dissolve it by ultrasound, let cool, pass through a D101 macroporous resin column (inner diameter 1.5 cm, column height 12 cm), elute with 50 mL of water, discard the water liquid, then elute with 30 mL of 40% ethanol, discard the eluate, and then elute with 80 mL of 70% ethanol, collect the eluate, evaporate it in a water bath, dissolve the residue in methanol, transfer it to a 5 mL measuring flask, add methanol to the scale, shake well, filter, and take the filtrate to obtain.

[0329] 4. Preparation of negative control solution

[0330] Take 2.0 g of negative extract powder without astragalus, weigh accurately, and prepare according to the "Preparation of test solution".

[0331] 5. Determination

[0332] Accurately pipette 10 μL and 20 μL of the reference solution, 20 μL of the test solution, and 20 μL of the negative control solution, respectively, and inject them into the high performance liquid chromatograph to determine the peak area and calculate the content using the external standard two-point method logarithmic equation.

[0333] Example 13.1 Specificity test

[0334] Accurately pipette 20 μL of reference solution, negative control solution and test solution respectively, inject them into high performance liquid chromatography instrument for determination.

[0335] See the results Figure 1-3 The negative control solution did not interfere with the detection of astragaloside IV in the extract powder sample. This method has good specificity for the determination of astragaloside IV.

[0336] Example 13.2 Instrument Precision Test

[0337] Accurately draw astragaloside IV reference solution (0.4524 mg / mL) and inject it into the high performance liquid chromatograph. Repeat the injection 6 times, 10 μL each time, measure the peak area, and calculate the RSD value.

[0338] The results are shown in Table 16. The instrument precision of this method is good.

[0339] Table 16 Instrument precision test results

[0340]

[0341] Example 13.3 Investigation of linear relationship

[0342] Accurately weigh 15.48 mg of astragaloside IV reference substance, place it in a 10 mL volumetric flask, add methanol to dissolve and dilute to the scale, shake well, and obtain 1.5078 mg mL -1 Solution ①: Accurately measure 5 mL of solution ① into a 10 mL volumetric flask, add methanol to the mark, and shake well to obtain 0.7539 mg mL -1 Solution ②: Accurately measure 3 mL of solution ② into a 5 mL volumetric flask, add methanol to the mark, and shake well to obtain 0.4524 mg mL -1 Solution ③: Accurately measure 2 mL of solution ③ into a 5 mL volumetric flask, add methanol to the mark, and shake well to obtain 0.3016 mg mL -1 Solution ④: Accurately measure 1 mL of solution ④ into a 5 mL volumetric flask, add methanol to the mark, and shake well to obtain 0.1508 mg mL -1 Solution ⑤: Accurately measure 1 mL of solution ⑤ into a 5 mL volumetric flask, add methanol to the mark, and shake well to obtain 0.0905 mg mL -1 Solution ⑥. Take 10 μL of each concentration of the reference solution and inject it into the HPLC. Measure the peak area of ​​the chromatographic peak and use the injection concentration (mg·mL-1 ) as the abscissa (X), the logarithm of the peak area as the ordinate (Y), and linear regression was performed. The results are shown in Table 17. Figure 4 .

[0343] The linear regression equation of astragaloside IV is: y = 1.6263x + 0.4041; R = 0.9992, and the range of astragaloside IV is 0.0905-1.5078 mg·mL -1 It showed good linearity within the concentration range.

[0344] Table 17 Results of investigation on the linear relationship of astragaloside IV

[0345]

[0346] Example 13.4 Solution stability test

[0347] Prepare an appropriate amount of astragaloside IV reference substance (Batch No. 110781-201616) according to the method under "Preparation of Reference Solution." Accurately pipette 10 μL of the reference solution and inject it into the HPLC at 0, 2, 4, 6, 12, 24, and 36 hours. Measure the peak area and calculate the content and RSD value. The results are shown in Table 18. Astragaloside IV is stable after 36 hours.

[0348] Table 18 Stability test results

[0349]

[0350] Example 13.5 Repeatability test

[0351] Take 2.0 g of dry extract powder (prepared according to the preparation method of Example 9, batch number: 2018091003) and prepare the test solution in 6 replicates according to the method under "Preparation of Test Solution". Accurately pipette 20 μL of each test solution and inject it into the high performance liquid chromatograph. Determine the peak area and calculate the content and RSD value. The results are shown in Table 19. This method has good reproducibility.

[0352] Table 19 Repeatability test results

[0353]

[0354] Example 13.6 Sample recovery test

[0355] Accurately weigh 6 samples of known content (prepared according to the preparation method of Example 9, batch number: 2018091003, astragaloside IV content: 1.03 mg g -1 ), 1 g of each portion was added to 8 mL of water, and 2 mL of astragaloside IV reference solution (0.4235 mg mL -1), prepare the test solutions according to the method under "Preparation of Test Solution", accurately pipette 20 μL of each test solution, inject into the HPLC, measure the peak area, and calculate the sample recovery. The results are shown in Table 20, and the sample recovery rate of this method is good.

[0356] Table 20 Sample recovery test results

[0357]

[0358] Example 13.7 Durability Test

[0359] 13.7.1 Column Temperature Investigation

[0360] The chromatographic conditions were the same except that the column temperature was changed to 25°C, 30°C, and 35°C. 2.0 g of dry extract powder (prepared according to the preparation method of Example 9, batch number: 2018091003) was accurately weighed and the test solution was prepared according to the method under "Preparation of Test Solution". 10 and 20 μL of the reference solution and 20 μL of the test solution were accurately pipetted into the HPLC instrument at different column temperatures. The content and RSD value were calculated. The results are shown in Table 21. This method has good durability at 25-35°C.

[0361] Table 21 Test results at different column temperatures

[0362]

[0363] 13.7.2 Investigation of Mobile Phase Ratio

[0364] The chromatographic conditions were the same as those described above, except that the mobile phase ratios were changed to 31:69, 32:68, and 33:67. 2.0 g of dry extract powder (prepared according to the preparation method of Example 9, batch number: 2018091003) was accurately weighed and the test solution was prepared according to the method under "Preparation of Test Solution." 10 and 20 μL of the reference solution and 20 μL of the test solution were accurately pipetted into the HPLC instrument at different mobile phase ratios. The contents and RSD values ​​were calculated. The results are shown in Table 22. This method demonstrated good robustness under different mobile phase ratios.

[0365] Table 22 Experimental results of different mobile phase ratios

[0366]

[0367] 13.7.3 Flow rate investigation

[0368] The chromatographic conditions were as follows except that the flow rate was changed to 0.8 ml min -1 、1.0ml·min -1 、1.2ml·min -1Other chromatographic conditions were the same as above. 2.0 g of dry extract powder (prepared according to the preparation method of Example 9, batch number: 2018091003) was accurately weighed and the test solution was prepared according to the method under "Preparation of Test Solution." 10 and 20 μL of the reference solution and 20 μL of the test solution were accurately pipetted into the HPLC instrument at different flow rates. The contents and RSD values ​​were calculated. The results are shown in Table 23. This method showed good robustness under different flow rates.

[0369] Table 23 Test results at different flow rates

[0370]

[0371] 13.7.4 Chromatographic Column Inspection

[0372] The chromatographic conditions were the same, except that the chromatographic column was changed to an SB-C18 column. 2.0 g of dry extract powder (prepared according to the preparation method in Example 9, batch number: 2018091003) was accurately weighed and the test solution was prepared according to the method under "Preparation of Test Solution." 10 and 20 μL of the reference solution and 20 μL of the test solution were accurately pipetted into the HPLC instrument, and the content and RSD values ​​were calculated. The results are shown in Table 24, demonstrating the robustness of this method under different chromatographic column conditions.

[0373] Table 24 Test results of different chromatographic columns

[0374]

[0375] Example 14: Detection of Chinese medicine composition samples

[0376] Three batches of dry extract powder (prepared according to the preparation method of Example 9, batch numbers: 2018091001, 2018091002, 2018091003) were taken, each 2.0 g, and two parallel portions were accurately weighed. Test solutions were prepared according to the method under "Preparation of Test Solution". 10 and 20 μL of the reference solution and 20 μL of the test solution were accurately aspirated and injected into the high performance liquid chromatograph. The content and RSD value were calculated. The results are shown in Table 25.

[0377] Table 25 Sample testing

[0378]

[0379] Example 15: Detection of Chinese medicine composition preparation samples

[0380] Grind an appropriate amount of SZ1105-SHP tablets (prepared according to the preparation method of Example 12, batch numbers: 20180110601, 2018110602, 2018110603) into a powder. Accurately weigh approximately 2.5 g of each powder in duplicate. Prepare test solutions according to the method under "Preparation of Test Solutions." Accurately pipette 10 and 20 μL of the reference solution and 20 μL of the test solution each into a high-performance liquid chromatograph, and calculate the content and RSD. The results are shown in Table 26.

[0381] Table 26 Sample test results

[0382]

[0383] The above further describes the present invention in conjunction with specific embodiments. However, these embodiments are merely exemplary and do not constitute any limitation on the scope of the present invention. It should be understood by those skilled in the art that the details and forms of the technical solutions of the present invention may be modified or replaced without departing from the spirit and scope of the present invention, and such modifications and replacements shall fall within the scope of protection of the present invention.

Claims

1. A Chinese medicine composition preparation for treating kidney disease, characterized in that: The invention is composed of a Chinese medicine composition and 1-90% of an excipient by weight; the Chinese medicine composition is made from the following raw materials: astragalus root, glossy privet fruit, white peony root, atractylodes macrocephala, honeysuckle, zedoaria and trigonosciadium japonicum.

2. The Chinese medicine composition preparation according to claim 1, characterized in that The excipients include at least one of pharmaceutically acceptable fillers, disintegrants, lubricants and coating excipients.

3. The Chinese medicine composition preparation according to claim 2, characterized in that The filler is at least one of microcrystalline cellulose, dextrin, and starch, preferably starch; the disintegrant is at least one of polyvinyl pyrrolidone, sodium carboxymethyl starch, starch, and low-substituted hydroxypropyl cellulose, preferably starch; the lubricant is at least one of talc, magnesium stearate, and micropowdered silica, preferably magnesium stearate; the coating excipient is at least one of hydroxypropyl methylcellulose, Eudragit E100, and Opadry 290W665025-CN, preferably Opadry 290W665025-CN; Preferably, the filler is starch, the disintegrant is starch, and the lubricant is magnesium stearate.

4. The Chinese medicine composition preparation according to claim 2, characterized in that The filler accounts for 15-25% of the weight of the traditional Chinese medicine composition preparation, the disintegrant accounts for 1-4% of the weight of the traditional Chinese medicine composition preparation, and the lubricant accounts for 0.1-0.5% of the weight of the traditional Chinese medicine composition preparation; preferably, the filler and disintegrant account for 19.9% ​​of the weight of the traditional Chinese medicine composition preparation, and the lubricant accounts for 0.1% of the weight of the traditional Chinese medicine composition preparation.

5. The Chinese medicine composition preparation according to claim 1, characterized in that The weight percentage of the excipient is 5-85%, preferably 10-80%, more preferably 15-75%, and most preferably 20%.

6. The Chinese medicine composition preparation according to claim 1, characterized in that The Chinese medicine composition is prepared from the following raw materials in parts by weight: 61-119 parts of Astragalus, 61-119 parts of Ligustrum lucidum, 37-71 parts of White Peony Root, 37-71 parts of Atractylodes Macrocephala, 80-105 parts of Honeysuckle, 49-95 parts of Curcuma and 49-95 parts of Trigonosciadium japonicum; Preferably, it is made from the following raw materials in parts by weight: 72-108 parts of Astragalus, 72-108 parts of Ligustrum lucidum, 42-66 parts of White Peony Root, 42-66 parts of Atractylodes Macrocephala, 80-105 parts of Honeysuckle, 54-90 parts of Curcuma and 54-90 parts of Trigonosciadium japonicum; More preferably, it is made from the following raw materials in parts by weight: 88-92 parts of Astragalus, 88-92 parts of Ligustrum lucidum, 52-55 parts of White Peony, 52-55 parts of Atractylodes macrocephala, 86-89 parts of Honeysuckle, 69-72 parts of Curcuma and 69-72 parts of Trigonosciadium japonicum, and further made from the following raw materials in parts by weight: 88-92 parts of Astragalus, 88-92 parts of Ligustrum lucidum, 52-55 parts of White Peony, 52-55 parts of Atractylodes macrocephala, 87.8 parts of Honeysuckle, 69-72 parts of Curcuma and 69-72 parts of Trigonosciadium japonicum; Most preferably, the medicine is prepared from the following raw materials in parts by weight: 90.1 parts of Astragalus, 90.1 parts of Ligustrum lucidum, 53.5 parts of White Peony Root, 53.5 parts of Atractylodes Macrocephala, 87.8 parts of Honeysuckle, 70.9 parts of Curcuma and 70.9 parts of Trigonosciadium japonicum.

7. The Chinese medicine composition preparation according to claim 1, characterized in that The preparation method of the Chinese medicine composition is: (1) Extracting volatile oil: extracting volatile oil from Atractylodes macrocephala and Curcuma zedoaria, and obtaining volatile oil, medicinal residue and extract I after solid-liquid separation; (2) Alcohol extraction: Astragalus and Ligustrum lucidum are extracted with alcohol, and solid-liquid separation is performed to obtain alcohol extract and alcohol-extracted medicinal residue; (3) Water extraction: extracting the medicinal residue obtained in step (1), the alcohol-extracted medicinal residue obtained in step (2), and the remaining raw materials by water extraction, and separating the solid and liquid to obtain aqueous extract II; (4) Concentration: The extract I obtained in step (1) and the aqueous extract II obtained in step (3) are combined and concentrated to obtain aqueous extract concentrate III; (5) alcohol precipitation: subjecting the aqueous extract concentrate III obtained in step (4) to alcohol precipitation to obtain an alcohol precipitation supernatant; (6) combining the alcohol extract obtained in step (2) and the alcohol precipitation supernatant obtained in step (5), and concentrating to obtain a thick paste; (7) The thick paste obtained in step (6) is dried and mixed with volatile oil to obtain; or (1) Soaking Atractylodes macrocephala and Curcuma zedoaria in water, extracting volatile oil, encapsulating, and drying to obtain an inclusion compound, and collecting the aqueous solution and the medicinal residue after extraction for later use; (2) extracting Astragalus and Ligustrum lucidum with ethanol, concentrating to obtain an alcohol extract, and collecting the alcohol-extracted medicinal residue for later use; (3) combining the alcohol-extracted medicinal residue obtained in step (2) and the medicinal residue obtained in step (1) with Trigonella ternate, white peony root, and honeysuckle, extracting with water, concentrating to obtain a water extract, precipitating with alcohol, and concentrating the supernatant to obtain an alcohol precipitate; (4) The alcohol extract obtained in step (2) and the alcohol precipitate obtained in step (3) are combined and dried, and then mixed with the inclusion compound obtained in step (1) to obtain the product.

8. The method for preparing the Chinese medicine composition according to any one of claims 1 to 7, characterized in that: The method comprises the following steps: uniformly mixing a traditional Chinese medicine composition with a filler and a disintegrant to prepare granules, adding a lubricant and tableting to obtain the traditional Chinese medicine composition preparation; preferably, after the tableting step, a coating step is further included, and the coating process is as follows: the solid content of the coating liquid is 15-20%, preferably 17%; the tablet bed temperature is 35-45°C, preferably 40°C; and the coating time is 50-60 minutes.

9. A method for controlling the quality of the Chinese medicine composition preparation according to any one of claims 1 to 7 or the Chinese medicine composition therein, comprising the following steps: S1. Preparation of test solution: dissolve the Chinese medicine composition preparation in water by ultrasonication, extract by shaking with water-saturated n-butanol, combine the n-butanol solutions, wash with ammonia test solution, discard the ammonia test solution, evaporate the n-butanol solution to dryness in a water bath, add water to the residue, dissolve it by ultrasonication, let it cool, pass it through a resin column, elute with water and ethanol respectively, discard the eluent, elute with ethanol again, collect the eluent, evaporate to dryness in a water bath, dissolve the residue in methanol, filter, and take the filtrate to obtain; S2. Detection: Detect the content of astragaloside IV using high performance liquid chromatography.

10. The quality control method according to claim 9, characterized in that: The resin in step S1 is a D101 macroporous resin, and the volume percentage of ethanol in step S1 is 30-80%; The detection conditions of the high performance liquid chromatography in step S2 are: octadecylsilane bonded silica gel as a filler, acetonitrile-water as a mobile phase, evaporative light scattering detector detection, a flow rate of 0.8-1.2 mL / min, and a column temperature of 25-35° C.; preferably, step S1 also includes the preparation of a reference solution: taking an astragaloside IV reference substance and adding methanol to prepare a solution; Step S1 also includes the preparation of a negative control solution: taking a negative extract that does not contain astragalus and preparing it according to the preparation method of the test solution.

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