Composition of traditional Chinese medicine extracts as well as preparation method and application of composition

Abstract

The invention discloses a traditional Chinese medicine extract composition as well as a preparation method and application thereof, and the traditional Chinese medicine extract composition comprises the following components in percentage by dry matter content of 100%: 1-10% of sennoside A, 1-10% of sennoside B, 1-10% of magnolol, 1-10% of honokiol, 30-50% of paeoniflorin, 5-10% of albiflorin, 0.1-0.5% of corydaline and 0.1-0.5% of tetrahydropalmatine. The raw medicinal materials comprise mangnolia officinalis, folium sennae, radix paeoniae alba (or radix paeoniae rubra) and rhizoma corydalis (or vinegar-processed rhizoma corydalis). The composition disclosed by the invention is rigorous in traditional Chinese medicine compatibility structure, ingenious in formula and unique in formula; the processing technology is advanced, green, environment-friendly and free of pollutants, medicine compound processing, medicine residue ecological utilization, quality control and reasonable development are integrated, the utilization rate is high, the purpose of the extract is special, and the market application prospect is wide.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of traditional Chinese medicine compound, and specifically discloses a composition of traditional Chinese medicine extract and a preparation method and application thereof. BACKGROUND

[0002] With the continuous changes of dietary structure, lifestyle, mental and psychological stress and social environmental factors, the incidence of chronic constipation shows a significant upward trend, involving various clinical fields, seriously disturbing people's daily life and affecting people's quality of life. Constipation is a common and frequently-occurring disease in China, usually characterized by less stool frequency (less than 3 times a week), dry and hard stool or non-dry stool but defecation disorder, among which, defecation disorder can also manifest as difficult defecation, defecation disorder, incomplete defecation, time-consuming defecation or the need for manual assistance for defecation. If the onset time is more than 6 months, chronic constipation can be diagnosed. Constipation is first mentioned in Huangdi Neijing, which is called "posterior incontinence" and "difficulty in urination and defecation"; in the Han Dynasty, Zhang Zhongjing called constipation "yang knot" and "yin knot" in Shanghan Zabing Lun, and "spleen knot" in Jingui Yaolue, and initially created "Maren Pill Prescription" to treat spleen knot syndrome, which is still a commonly used prescription for treating constipation today.

[0003] Abdominal pain refers to the pain in the abdominal region caused by various reasons, which is the manifestation of the lesions of internal and external organs in the abdominal cavity. The causes of acute and chronic abdominal pain are numerous, including diseases of internal organs in the abdominal cavity, diseases of the abdominal wall, diseases of the chest cavity, toxic and metabolic disorders, organic neuropathy and systemic diseases, etc. Constipation accompanied by severe abdominal pain, abdominal distension and vomiting often suggests acute constipation, abdominal gas obstruction and intestinal stasis.

[0004] Due to the complexity of traditional Chinese medicine ingredients, mainly including lignans, glycosides and alkaloids, etc., these active ingredients help to lubricate the intestines, relieve abdominal pain and other effects. Under the guidance of traditional Chinese medicine theory, for the same indications, different dialectical mechanisms, the choice of prescription is different, and the choice and dosage of the original medicinal materials are also different.

[0005] Irritable bowel syndrome (IBS) is a functional disease characterized by abdominal pain and changes in bowel habits. Globally, the prevalence of IBS is more than 11%, and about one-third of IBS patients have constipation as the main symptom (IBS-C), which is characterized by abdominal pain, reduced defecation frequency and difficulty in defecation. Clinical treatment includes lifestyle adjustment and drug therapy. However, these methods can only temporarily relieve specific symptoms and cannot maintain long-term efficacy, leading to poor patient compliance and increased treatment costs. Therefore, it is crucial to develop new therapeutic drugs. SUMMARY

[0006] The purpose of the present application is to provide a composition of traditional Chinese medicine extract and a preparation method and application thereof.

[0007] In one aspect, the present application provides a composition of traditional Chinese medicine extract, wherein, based on 100% of dry matter content of the composition, it contains: sennoside A 1-5%, sennoside B 1-5%, magnolol 1-5%, honokiol 1-5%, paeoniflorin 30-50%, albiflorin 5-10%, fumarine 0.1-0.3%, and stylopine 0.1-0.3%, and the rest is other substances.

[0008] According to a specific embodiment of the present application, preferably, based on 100% of dry matter content of the composition of traditional Chinese medicine extract, it contains: sennoside A 1-5%, sennoside B 1-5%, magnolol 1-5%, honokiol 1-5%, paeoniflorin 30-50%, albiflorin 5-10%, fumarine 0.1-0.3%, and stylopine 0.1-0.3%, and the rest is other substances.

[0009] According to a specific embodiment of the present application, preferably, based on 100% of dry matter content of the composition of traditional Chinese medicine extract, it contains: sennoside A 1-5%, sennoside B 1-5%, magnolol 1-5%, honokiol 1-5%, paeoniflorin 30-50%, albiflorin 5-10%, fumarine 0.1-0.3%, and stylopine 0.1-0.3%, and the rest is other substances.

[0010] In another aspect, the present application provides a composition of traditional Chinese medicine extract, wherein the composition of traditional Chinese medicine extract comprises extracts of original medicinal materials; the original medicinal materials comprise Magnolia officinalis, Folium Sennae, Radix Paeoniae Alba, and Sargentcolumbofis Rhizoma; and the mass ratio of the original medicinal materials Magnolia officinalis, Folium Sennae, Radix Paeoniae Alba, and Sargentcolumbofis Rhizoma is (1-20):(1-30):(1-25):(5-50).

[0011] Preferably, the mass ratio of the original medicinal materials Magnolia officinalis, Folium Sennae, Radix Paeoniae Alba, and Sargentcolumbofis Rhizoma is (10-20):(1-10):(15-25):(5-15) or (1-2):(1-10):(1-10):(20-50); preferably, the mass ratio of the original medicinal materials Magnolia officinalis, Folium Sennae, Radix Paeoniae Alba, and Sargentcolumbofis Rhizoma is (10-20):(1-5):(15-25):(8-15); and more preferably, the mass ratio of the original medicinal materials Magnolia officinalis, Folium Sennae, Radix Paeoniae Alba, and Sargentcolumbofis Rhizoma is 15:2:20:10.

[0012] The composition of the present application can be composed of concentrated Chinese medicine granules (CCMG) containing equal amounts of original medicinal materials.

[0013] The composition of the present application can also be composed of purified Chinese Medicine extracts (PCME) containing equivalent amount of Q-markers of raw medicinal materials.

[0014] The information of medicinal materials contained in the Chinese medicine composition of the present application is shown in the following table:

[0015]

[0016] In another aspect, the present application provides a pharmaceutical composition for treating constipation, which is composed of the above-mentioned composition of Chinese medicine extracts and one or more pharmaceutically acceptable excipients.

[0017] According to a specific embodiment of the present application, preferably, the dosage form of the pharmaceutical composition comprises one or a combination of two or more of a capsule, a tablet, a powder, a solution or a suspension.

[0018] In another aspect, the present application provides a preparation method of the above-mentioned composition of Chinese medicine extracts or the above-mentioned pharmaceutical composition, wherein the preparation method comprises:

[0019] The composition of Chinese medicine extracts or the pharmaceutical composition is obtained by mixing Magnolia officinalis extract, senna leaf extract, Radix Paeoniae Alba or Rubra extract, Corydalis or acetic Corydalis extract.

[0020] According to a specific embodiment of the present application, the preparation method of Magnolia officinalis extract comprises: extracting Magnolia officinalis with water or ethanol to obtain Magnolia officinalis extract solution, further concentrating to obtain Magnolia officinalis extract concentrate, and extracting the Magnolia officinalis extract concentrate with petroleum ether to obtain Magnolia officinalis extract.

[0021] According to a specific embodiment of the present application, the preparation method of senna leaf extract comprises: extracting senna leaf with water, sodium bicarbonate or ethanol to obtain senna leaf extract solution, further concentrating to obtain senna leaf concentrate, and purifying the senna leaf concentrate by column to obtain senna leaf extract.

[0022] According to a specific embodiment of the present application, the preparation method of Radix Paeoniae Alba or Rubra extract comprises: extracting Radix Paeoniae Alba or Rubra with water or ethanol to obtain Radix Paeoniae Alba or Rubra extract solution, further concentrating to obtain Radix Paeoniae Alba or Rubra concentrate, and purifying the Radix Paeoniae Alba or Rubra concentrate by column to obtain Radix Paeoniae Alba or Rubra extract.

[0023] According to a specific embodiment of the present application, the preparation method of Corydalis or acetic Corydalis extract comprises: extracting Corydalis or acetic Corydalis with water or ethanol to obtain Corydalis or acetic Corydalis extract solution, further concentrating to obtain Corydalis or acetic Corydalis concentrate, and purifying the Corydalis or acetic Corydalis concentrate by column to obtain Corydalis or acetic Corydalis extract.

[0024] According to the specific embodiment of the present application, the temperature for water or ethanol extraction in the preparation method of the Magnoliae officinalis extract is 40-90°C water; and the temperature for petroleum ether extraction is 40-70°C.

[0025] According to the specific embodiment of the present application, the temperature for water, sodium bicarbonate or ethanol extraction in the preparation method of the Folii sennae extract is 25-90°C.

[0026] According to the specific embodiment of the present application, the temperature for water or ethanol extraction in the preparation method of the Radix paeoniae alba or Radix paeoniae rubra extract is 60-90°C.

[0027] According to the specific embodiment of the present application, the temperature for water or ethanol extraction in the preparation method of the Rhizoma corydalis or Rhizoma corydalis vinegar extract is 60-90°C.

[0028] According to the specific embodiment of the present application, preferably, the preparation method of the Magnoliae officinalis extract comprises:

[0029] (1) crushing Magnoliae officinalis, adding water or 30%-95% ethanol in an amount of 8-12 times, and extracting at 40-90°C water bath reflux for 1-3 times, each time for 0.5-1.5 hours to obtain Magnoliae officinalis extract liquid;

[0030] (2) filtering and combining the Magnoliae officinalis extract liquid to obtain filtrate; and concentrating the filtrate at 40-80°C under reduced pressure to Magnoliae officinalis extract paste with a relative density of 1.15-1.20;

[0031] (3) adding petroleum ether in an amount of 50-150 times to the Magnoliae officinalis extract paste, and extracting at 40-70°C water bath reflux for 3-12 hours to obtain Magnoliae officinalis petroleum ether extract liquid;

[0032] (4) precipitating crystals by cooling the Magnoliae officinalis petroleum ether extract liquid, evaporating the petroleum ether at 40-70°C, and freeze-drying to obtain Magnoliae officinalis extract.

[0033] According to the specific embodiment of the present application, preferably, the preparation method of the Folii sennae extract comprises:

[0034] (1) adding water, 0.1% sodium bicarbonate or 30%-95% ethanol in an amount of 80-120 times to Folii sennae, and extracting at 25-90°C water bath reflux for 1-3 times, each time for 0.5-1.5 hours to obtain Folii sennae extract liquid;

[0035] (2) filtering and combining the Folii sennae extract liquid to obtain filtrate; and concentrating the filtrate at 40-80°C under reduced pressure to Folii sennae concentrated liquid with a relative density of 1.05-1.10;

[0036] (3) The senna leaf concentrated solution is purified by column, eluted with 5-10 times column volume of 5% ethanol to obtain an eluate, and then freeze-dried to obtain the senna leaf extract.

[0037] According to the specific embodiments of the present application, preferably, the preparation method of the Radix Paeoniae Alba or Rubra extract comprises:

[0038] (1) The Radix Paeoniae Alba or Rubra is crushed, 8-12 times amount of water or 30%-95% ethanol is added, and the mixture is refluxed in a water bath at 60-90°C for 1-3 times, each time for 0.5-1.5 hours to obtain a Radix Paeoniae Alba or Rubra extract solution;

[0039] (2) The Radix Paeoniae Alba or Rubra extract solution is filtered and combined, and the filtrate is concentrated under reduced pressure at 40-80°C to a Radix Paeoniae Alba or Rubra concentrated solution with a relative density of 1.05-1.10;

[0040] (3) The Radix Paeoniae Alba or Rubra concentrated solution is purified by column, eluted with 8-16 times column volume of 20% ethanol to obtain an eluate, and then freeze-dried to obtain the Radix Paeoniae Alba or Rubra extract.

[0041] According to the specific embodiments of the present application, preferably, the preparation method of the Radix Paeoniae Alba or Rubra extract comprises:

[0042] (1) The Radix Paeoniae Alba or Rubra is crushed, 8-12 times amount of water or 30%-95% ethanol is added, and the mixture is refluxed in a water bath at 60-90°C for 1-3 times, each time for 0.5-1.5 hours to obtain a Radix Paeoniae Alba or Rubra extract solution;

[0043] (2) The Radix Paeoniae Alba or Rubra extract solution is filtered and combined, and the filtrate is concentrated under reduced pressure at 40-80°C to a Radix Paeoniae Alba or Rubra concentrated solution with a relative density of 1.05-1.10;

[0044] (3) The Radix Paeoniae Alba or Rubra concentrated solution is purified by column, eluted with 8-16 times column volume of 20% ethanol to obtain an eluate, and then freeze-dried to obtain the Radix Paeoniae Alba or Rubra extract.

[0045] According to the specific embodiments of the present application, the preparation method further comprises a step of detecting the active ingredients in the above-mentioned traditional Chinese medicine extract composition or the above-mentioned pharmaceutical composition.

[0046] According to the specific embodiments of the present application, the active ingredients include magnolol, honokiol, sennoside A, sennoside B, paeoniflorin, albiflorin, stylopine, and tetrahydropalmatine.

[0047] According to the specific embodiments of the present application, preferably, the detection is liquid chromatography-tandem mass spectrometry detection.

[0048] According to a specific embodiment of the present application, preferably, in the liquid chromatography-tandem mass spectrometry detection, the chromatographic conditions comprise:

[0049] The chromatographic column is filled with octadecylsilane-bonded silica gel, and the theoretical plate number of the chromatographic column is ≥1000; the column temperature is 40℃;

[0050] The mobile phase comprises mobile phase A and mobile phase B, the mobile phase A is 0.1% formic acid aqueous solution, and the mobile phase B is 0.1% formic acid acetonitrile solution;

[0051] The elution mode is gradient elution, and the elution program is as follows:

[0052]

[0053]

[0054] The mass spectrometry conditions comprise:

[0055] A triple quadrupole mass spectrometry detector;

[0056] Electrospray ionization in negative mode;

[0057] The detection mode is multiple reaction monitoring;

[0058] When detecting magnolol, the multiple reaction monitoring ion pairs are m / z 265.0→247.1 and m / z 265.0→245.0;

[0059] When detecting honokiol, the multiple reaction monitoring ion pairs are m / z 265.0→249.0 and m / z 265.0→223.0;

[0060] When detecting sennoside A and sennoside B, the multiple reaction monitoring ion pairs are m / z 861.4→699.0 and m / z 861.4→386.0;

[0061] When detecting paeoniflorin, the multiple reaction monitoring ion pairs are m / z 525.0→327.0 and m / z 525.0→121.0;

[0062] When detecting paeoniflorin lactone, the multiple reaction monitoring ion pairs are m / z 525.0→479.0 and m / z 525.0→121.0;

[0063] When detecting fumaritine, the multiple reaction monitoring ion pairs are m / z 356.1→159.0 and m / z 356.1→159.0;

[0064] When detecting fumaritin, the multiple reaction monitoring ion pairs are m / z 370.2→170.0 and m / z 370.2→170.0.

[0065] In another aspect, the present application provides a use of the above-mentioned traditional Chinese medicine extract, the above-mentioned pharmaceutical composition or the above-mentioned preparation method in the preparation of a drug for preventing and / or treating constipation.

[0066] According to a specific embodiment of the present application, preferably, the constipation is abdominal pain type constipation.

[0067] The present application discloses a compound composition for treating abdominal pain type constipation and a preparation process thereof. The compound composition is prepared from Magnolia officinalis, senna leaf, Radix Paeoniae Alba (or Radix Paeoniae Rubra) and Rhizoma Corydalis (or Rhizoma Corydalis in vinegar). The present application also provides a preparation method and a quality control method of the compound composition. The compound composition has a strict medicinal compatibility structure, a clever prescription, a reliable prescription and a unique prescription. The processing technology is advanced, green, pollution-free, and integrates compound processing, ecological utilization of residues, quality control and reasonable development. The utilization rate is high, the extract has a special purpose, and the market application prospect is broad. The compound composition has a significant therapeutic effect on the prevention and treatment of IBS-C, has no obvious toxic and side effects, and has the advantages of green health. BRIEF DESCRIPTION OF DRAWINGS

[0068] Figure 1 Magnolia officinalis standard curve in Example 5.

[0069] Figure 2 Magnolia officinalis standard curve in Example 5.

[0070] Figure 3 Sennoside A standard curve in Example 5.

[0071] Figure 4 Sennoside B standard curve in Example 5.

[0072] Figure 5 Paeoniflorin standard curve in Example 5.

[0073] Figure 6 Paeoniflorin standard curve in Example 5.

[0074] Figure 7 Tetrahydropalmatine standard curve in Example 5.

[0075] Figure 8 Tetrahydropalmatine standard curve in Example 5.

[0076] Figure 9 The effect of four optional compositions of CDD-2105 on the defecation frequency of loperamide-induced constipation mice within 6h.

[0077] Figure 10 The effect of four optional compositions of CDD-2105 on the fecal water content of loperamide-induced constipation mice.

[0078] Figure 11 Effect of CDD-2105 purified Chinese medicine extract (PCME) on the feces water content of loperamide-induced constipation mice.

[0079] Figure 12 Effect of CDD-2105 purified Chinese medicine extract (PCME) on the feces water content of loperamide-induced constipation mice.

[0080] Figure 13 Effect of four optional compositions of CDD-2105 on the latency of mice's response to thermal stimulation-induced pain.

[0081] Figure 14 Effect of CDD-2105 purified Chinese medicine extract (PCME) on the latency of mice's response to thermal stimulation-induced pain.

[0082] Figure 15 Effect of CDD-2105 Chinese medicine quality marker mixture (MM) on the body weight change of mice.

[0083] Figure 16 Effect of CDD-2105 Chinese medicine quality marker mixture (MM) on the food intake of mice.

[0084] Figure 17 Effect of CDD-2105 Chinese medicine quality marker mixture (MM) on the organ coefficient of mice. DETAILED DESCRIPTION

[0085] The present application will be further described in detail by specific examples, but the protection scope of the present application is not limited. Unless otherwise specified, the experimental methods used in the present application are conventional methods, and the experimental apparatuses, materials, reagents, etc. used can be obtained from commercial channels.

[0086] I. Production and quality control

[0087] 1. Instruments

[0088] Ultrasonic cleaner (KQ-500DE, Kunshan Ultrasonic Instrument Co., Ltd.); ultra-high performance liquid chromatography coupled with triple quadrupole mass spectrometer (Agilent 1290 Infinity II-Agilent 6470 LC / TQ, Agilent Technologies, USA); electronic analytical balance (ml204 / 02, Mettler-Toledo); electronic analytical balance (XPR205, Mettler-Toledo); electronic analytical balance (E600-2, Changshu Shuangjie Testing Instrument Factory).

[0089] 2. Materials and reagents

[0090] Magnolia officinalis (Batch No. 20221014-13, Shanghai Hunliang Standard Technology Service Co., Ltd.); Senna leaf (Batch No. 20221014-02, Shanghai Hunliang Standard Technology Service Co., Ltd.); Radix Paeoniae Alba (Batch No. 20221014-07, Shanghai Hunliang Standard Technology Service Co., Ltd.); Radix Paeoniae Rubra (Batch No. 20221014-08, Shanghai Hunliang Standard Technology Service Co., Ltd.); Rhizoma Corydalis in vinegar (Batch No. 2210006-ZJ, Sichuan Xinhua Traditional Chinese Medicine Co., Ltd.); Corydalis tuber (Batch No. 20221014-10, Shanghai Hunliang Standard Technology Service Co., Ltd.); Magnolol (Batch No. 110729-202015, China National Institute for Food and Drug Control); and honokiol (Batch No. YY90181, Shanghai Yuyeh Co., Ltd.); Sennoside A (Batch No. CFS202201, Chemfaces); Sennoside B (Batch No. CFS202102, Chemfaces); Paeoniflorin (Batch No. NO811A, Meilunbio); Albiflorin (Batch No. B21149, Shanghai Yuyeh Co., Ltd.); Tetrahydropalmatine (Batch No. B74569, Shanghai Yuyeh Co., Ltd.); Corydaline (Batch No. B21425, Shanghai Yuyeh Co., Ltd.).

[0091] Acetonitrile (OMINI) was chromatographically pure, methanol (Guangdong Guanghua Science and Technology Co., Ltd.) was analytically pure, and water was ultrapure water.

[0092] Example 1

[0093] Magnolia officinalis was extracted and purified as follows:

[0094] 15 g of Magnolia officinalis was ground (through a 100-mesh screen), 95% ethanol was added in an amount of 12 times, and water bath reflux extraction was performed for 3 times, 1.5 hours each time. The extraction liquid was filtered and combined, and the filtrate was concentrated under reduced pressure (40-80°C) to a relative density of 1.15-1.20 of the extract. Petroleum ether was added in an amount of 150 times, and extraction was performed at 40°C water bath for 12 hours. The petroleum ether extract was separated while hot, and was left to stand (4°C) overnight to precipitate crystals. The petroleum ether was evaporated (40-70°C), and the Magnolia officinalis dry extract powder was obtained by freeze-drying. The yield was 4.91%, the purity was 81.17%, the Magnolol content was 37.95%, and the Honokiol content was 43.22%.

[0095] Example 2

[0096] Senna leaf was extracted and purified as follows:

[0097] 2g senna leaf without crushing, 80 times the amount of water, water bath reflux extraction 3 times, 30 minutes each time, the extract was filtered and combined, the filtrate was concentrated under reduced pressure (40℃-80℃) to a concentrated solution with a relative density of 1.05-1.10. Prepare AB-8 macroporous resin, column packing, wet sample loading. First use 5 times the column volume of water and 8 times the column volume of 0.5% NaHCO3 to remove impurities, then use 10 times the column volume of 5% ethanol to elute and collect the fractions, and freeze-dry to obtain senna leaf dry extract powder, with a yield of 1.53% and a purity of 29.87%, of which sennoside A content is 14.71%, sennoside B content is 15.15%;

[0098] Example 3

[0099] The white peony root was extracted and purified as follows:

[0100] 20g white peony root was crushed (through a 100 mesh screen), 30% ethanol was added 12 times the amount, and water bath reflux extraction was performed 3 times, 30 minutes each time. The extract was filtered and combined, and the filtrate was concentrated under reduced pressure (40℃-80℃) to a concentrated solution with a relative density of 1.05-1.10. Prepare AB-8 macroporous resin, column packing, wet sample loading. First use 8 times the column volume of water to remove impurities, then use 16 times the column volume of 20% ethanol to elute and collect the fractions, and freeze-dry to obtain white peony root dry extract powder, with a yield of 6.0% and a purity of 81.96%, of which paeoniflorin content is 53.35%, paeonolactone glycoside content is 28.61%;

[0101] Example 4

[0102] 10g of the vinegar fuzi was extracted and purified as follows:

[0103] The vinegar fuzi was crushed (through a 100 mesh screen), 50% ethanol was added 10 times the amount, and water bath reflux extraction was performed 3 times, 30 minutes each time. The extract was filtered and combined, and the filtrate was concentrated under reduced pressure (40℃-80℃) to a concentrated solution with a relative density of 1.05-1.10. Prepare AB-8 macroporous resin, column packing, wet sample loading. First use 8 times the column volume of water and 8 times the column volume of 40% ethanol to remove impurities, then use 8 times the column volume of 95% ethanol to elute and collect the fractions, and freeze-dry to obtain vinegar fuzi dry extract powder, with a yield of 0.187% and a purity of 14.58%, of which tetrahydropalmatine content is 6.57%, and benzylisoquinoline alkaloids content is 8.01%;

[0104] Example 5

[0105] I. 20mg, 69.83mg, 374mg, 12.48mg of the magnolia officinalis dry extract powder, senna leaf dry extract powder, white peony root dry extract powder, and vinegar fuzi dry extract powder obtained in Examples 1-4 were mixed to obtain a refined extract composition (Purified Chinese Medicine extracts, PCME).

[0106] II. Quality control method of composition index component, specifically comprising:

[0107] The composition is subjected to content determination by liquid chromatography-mass spectrometry, specifically as follows:

[0108] Chromatographic conditions and system suitability test:

[0109] Octadecylsilane-bonded silica gel is used as the filler; 0.1% formic acid aqueous solution is used as the mobile phase A, and 0.1% formic acid acetonitrile solution is used as the mobile phase B, which is subjected to gradient elution according to the following procedure:

[0110] Time (min) Mobile phase A (%) Mobile phase B (%) 0-6 88→75 12→25 6-7 75→55 25→45 7-10 55→20 45→80 10-10.1 20→0 80→100

[0111] 1. Magnolol and honokiol

[0112] The chromatographic conditions described in the "chromatographic conditions and system suitability test" are used, and the column temperature is maintained at 40℃. A triple quadrupole mass spectrometer detector and an electro-spray ionization negative mode (ESI - ) are used, and the multiple reaction monitoring (MRM) ion pairs m / z 265.0→247.1 and m / z 265.0→245.0 are used. The theoretical plate number of the chromatographic column is not less than 1000, and the reference magnolol peak is calculated.

[0113] The column temperature is maintained at 40℃. A triple quadrupole mass spectrometer detector and an electro-spray ionization negative mode (ESI - ) are used, and the multiple reaction monitoring (MRM) ion pairs m / z 265.0→249.0 and m / z 265.0→223.0 are used. The theoretical plate number of the chromatographic column is not less than 1000, and the reference honokiol peak is calculated.

[0114] The reference solution is prepared by accurately weighing a certain amount of magnolol and honokiol reference substances, adding methanol to prepare a series of standard solutions containing 6.0, 15.0, 30.0, 60.0, and 90.0 μg of magnolol and honokiol per 1 ml, respectively. Figure 1 The standard curve of magnolol is prepared; Figure 2 The standard curve of honokiol is prepared.

[0115] Preparation of test sample solution

[0116] About 0.2 g of the dry extract powder of the composition is accurately weighed, placed in a conical flask with a stopper, 25 ml of methanol is accurately added, soaked for 24 hours, filtered, and the filtrate is obtained.

[0117] Determination method

[0118] 1 μl of the reference solution and the test sample solution is accurately taken respectively, injected into the liquid chromatograph, and determined, and the results are obtained.

[0119] 2. Sennoside A and Sennoside B

[0120] The column temperature was maintained at 40 °C using the chromatographic conditions described in the "Chromatographic conditions and system suitability test". A triple quadrupole mass spectrometer detector and electrospray ionization in negative mode (ESI - ) was used with multiple reaction monitoring (MRM) ion pairs m / z 861.4→699.0 and m / z 861.4→386.0. The theoretical plates of the column were not less than 1000, calculated with reference to the sennoside A peak and the sennoside B peak.

[0121] Preparation of the reference solution The sennoside A and sennoside B reference substances were weighed accurately and dissolved in 0.1% NaHC03 solution to obtain a series of standard solutions containing 6.0, 15.0, 30.0, 60.0, 90.0 μg of sennoside A and sennoside B per 1 ml, respectively. Figure 3 Sennoside A standard curve in Example 5; Figure 4 Sennoside B standard curve in Example 5.

[0122] Preparation of the test solution

[0123] About 0.5 g of the composition dry extract powder was accurately weighed into a conical flask with a stopper, 50 ml of 0.1% NaHC03 solution was accurately added, the weight was determined, ultrasonic treatment (power 250 W, frequency 40 kHz) was carried out for 15 minutes, it was removed and allowed to cool, the weight was again determined, the lost weight was made up with 0.1% NaHC03 solution, it was shaken well, filtered and the filtrate was obtained.

[0124] Determination method

[0125] 2 μl of the reference solution and the test solution were accurately pipetted respectively and injected into the liquid chromatograph, and determination was carried out, and the test solution was obtained.

[0126] 3. Paeoniflorin and Albiflorin

[0127] The column temperature was maintained at 40 °C using the chromatographic conditions described in the "Chromatographic conditions and system suitability test". A triple quadrupole mass spectrometer detector and electrospray ionization in negative mode (ESI - ) was used with multiple reaction monitoring (MRM) ion pairs m / z 525.0→327.0 and m / z 525.0→121.0. The theoretical plates of the column were not less than 1000, calculated with reference to the paeoniflorin peak.

[0128] The column temperature was maintained at 40 °C. A triple quadrupole mass spectrometer detector and electrospray ionization in negative mode (ESI -), using multiple reaction monitoring (MRM) ion pairs m / z 525.0→ 479.0 and m / z 525.0→ 121.0. The theoretical plates of the column are not less than 1000, calculated with reference to the peak of paeonol glycoside.

[0129] The preparation of the control solution: accurately weigh appropriate amount of paeonol and paeonol glycoside reference substances, add methanol to prepare a series of standard solutions containing 6.0, 15.0, 30.0, 60.0, 90.0 μg of paeonol and paeonol glycoside per 1 ml, respectively. Figure 5 The standard curve of paeonol is as follows: Figure 6 The standard curve of paeonol glycoside is as follows.

[0130] Preparation of test solution

[0131] Accurately weigh about 0.5 g of the composition dry extract powder, place it in a conical flask with a stopper, accurately add 25 ml of methanol, weigh, soak for 4 hours, then ultrasonically treat (power 250 W, frequency 40 kHz) for 20 minutes, take it out, cool, re-weigh, make up the weight loss with methanol, shake well, filter, and take the filtrate, which is the test solution.

[0132] Determination method

[0133] Accurately pipette 1 μl of the control solution and the test solution, respectively, into the liquid chromatograph, and determine, to obtain the results.

[0134] 4. Corydalis B and Corydalis A

[0135] The chromatographic conditions described in the "Chromatographic conditions and system suitability test" are used, and the column temperature is maintained at 40℃. A triple quadrupole mass spectrometer detector and electrospray ionization positive mode (ESI + ) are used, and multiple reaction monitoring (MRM) ion pairs m / z 356.1→ 159.0 and m / z 356.1→ 159.0 are used. The theoretical plates of the column are not less than 1000, calculated with reference to the peak of corydalis B.

[0136] The column temperature is maintained at 40℃. A triple quadrupole mass spectrometer detector and electrospray ionization positive mode (ESI + ) are used, and multiple reaction monitoring (MRM) ion pairs m / z 370.2→ 170.0 and m / z 370.2→ 170.0 are used. The theoretical plates of the column are not less than 1000, calculated with reference to the peak of corydalis A.

[0137] The preparation of the control solution: accurately weigh appropriate amount of corydalis B and corydalis A reference substances, add methanol to prepare a series of standard solutions containing 6.0, 15.0, 30.0, 60.0, 90.0 μg of corydalis B and corydalis A per 1 ml, respectively. Figure 7 The standard curve of corydalis B is as follows:Figure 8 CDD-2105 standard curve.

[0138] Preparation of test solution

[0139] About 0.5 g of the dry powder of the composition was precisely weighed into a conical flask with a stopper, 50 ml of concentrated ammonia-methanol (1:20) mixed solution was precisely added, the weight was determined, and after cold soaking for 1 hour, it was refluxed in a water bath at 80°C for 1 hour, taken out, cooled, and weighed again. The lost weight was made up with concentrated ammonia-methanol (1:20) mixed solution, shaken well, filtered, and the filtrate was evaporated in an evaporating dish. Methyl alcohol was added for dissolution, and then transferred to a 5 ml volumetric flask. Methyl alcohol was added to the mark of the volumetric flask. Shake well, filter, and use the continuous filtrate as the test solution.

[0140] Determination method

[0141] 1 μl of the control solution and the test solution was precisely taken and injected into the liquid chromatograph, and the determination was performed, and the results were obtained.

[0142] 5. The content determination results are shown in Table 1:

[0143] Table 1

[0144] Index component Dry paste powder content (%) Sennoside A 1.6539 Sennoside B 2.2611 Magnolol 2.2408 Honokiol 1.6849 Paeoniflorin 42.7401 Albiflorin 6.3684 Fuzhengsu B 0.1748 Fuzhengsu A 0.1407

[0145] II. Pharmacological activity experiment

[0146] Materials and methods

[0147] 1. Materials

[0148] CDD-2105: CDD-2105 composition solutions were prepared using 0.5% carboxymethylcellulose sodium (CMC-Na) solution, including CDD-2105 formula granule composition (Concentrated Chinese Medicine Granules, CCMG), CDD-2105 purified extract composition (Purified Chinese Medicine extracts, PCME), and CDD-2105 traditional Chinese medicine quality marker composition (Q-markers Mixture, MM). The intragastrically administered volume of mice was 15 ml / kg.

[0149] Loperamide solution: Loperamide solution was prepared at a concentration of 0.5 mg / ml, and the intragastrically administered amount of mice was 5 ml / kg.

[0150] Linaclotide solution: Linaclotide solution was prepared at a concentration of 3.97 μg / ml using 0.5% CMC-Na solution, and the intragastrically administered amount of mice was 15 ml / kg.

[0151] 2. Method

[0152] 2.1 Evaluation of the laxative effect of CDD-2105 on loperamide-induced constipation mice

[0153] (1) Animals and administration

[0154] Experiment 1: 6-8 week-old C57BL / 6J male mice were randomly divided into 6 groups, 8 mice in each group, namely normal control group (Control), loperamide group (Lop), Lop+CDD-2105 CCMG_1, Lop+CDD-2105 CCMG_2, Lop+CDD-2105 CCMG_3 and Lop+CDD-2105 CCMG_4. All CDD-2105 treatment groups were given CDD-2105 CCMG by gavage. The amount of crude drugs contained in CDD-2105 traditional Chinese medicine formula granule composition is shown in Table 2, and the administration of each group is shown in Table 3.

[0155] Table 2

[0156]

[0157] Table 3

[0158]

[0159] Experiment 2: 6-8 week-old C57BL / 6J male mice were randomly divided into 6 groups, 10 mice in each group, namely normal control group (Control), loperamide group (Lop), Lop+CDD-2105 CCMG group, Lop+CDD-2105 PCME (0.25x) group, Lop+CDD-2105 PCME (0.5x) group and Lop+CDD-2105 PCME (1x) group. The amount of crude drugs contained in CDD-2105 CCMG was the same as that in CDD-2105 CCMG_4, and the content of traditional Chinese medicine quality markers in each composition is shown in Table 4.

[0160] Table 4

[0161]

[0162] (2) Administration method: On the day of the experiment, except for the normal control group of mice given Mili-Q water by gavage, the rest of the mice were given loperamide by gavage, and each test drug was given at the same time.

[0163] (3) Defecation frequency: On the day of the experiment, the number of fecal particles within 6 hours after administration was recorded as the defecation frequency.

[0164] (4) Water content: On the day of the experiment, fecal samples from mice within 6 hours after administration were collected in a 2ml centrifuge tube with a known weight, the wet weight of the feces was recorded, and after drying at 105°C overnight, the dry weight of the feces was recorded, the water content of the mouse feces was calculated and compared.

[0165] (5) Results

[0166] From Figure 9 , Figure 10 it can be seen that compared with the model group, CDD-2105CCMG_2, _3, _4 significantly accelerated the defecation frequency of mice with constipation induced by loperamide, and increased the water content of feces. Because the diarrhea of CDD-2105CCMG_2, _3 group was more serious, the content of traditional Chinese medicine quality markers in CDD-2105PCME(1x) was set to be equivalent to that in CDD-2105CCMG_4 in the follow-up experiment. The results showed that CDD-2105PCME(1x) significantly increased the defecation frequency of mice with constipation induced by loperamide, and the effect was equivalent to that of CDD-2105CCMG( Figure 11 ). Except for CDD-2105PCME(0.25x), other doses of CDD2105 PCME significantly increased the water content of feces of mice with constipation induced by loperamide( Figure 12 ). These results showed that CDD-2105 can effectively relieve the constipation symptoms of mice induced by loperamide.

[0167] 2.2 Evaluation of the analgesic effect of CDD-2105 on mice with central pain induced by heat stimulation

[0168] (1) Animals and administration

[0169] Experiment one: 6-8 week old C57BL / 6J male mice, the baseline pain threshold of mice was measured before the experiment, and mice with pain threshold in the range of 5s-30s were selected, and qualified mice were randomly divided into normal control group, CDD-2105CCMG_1 group, CDD-2105CCMG_2 group, CDD-2105CCMG_3 group, CDD2105 CCME_4 group, 8 mice in each group. The amount of crude drug contained in CDD-2105 groups is shown in Table 2, and the administration of each group is shown in Table 5.

[0170] Table 5

[0171] Group Dosing condition Control group 0.5% CMC-Na, 15 ml / kg, i.g. CDD-2105CCMG_1 CDD-2105CCMG_1, 15 ml / kg, i.g. CDD-2105CCMG_2 CDD-2105CCMG_2, 15 ml / kg, i.g. CDD-2105CCMG_3 CDD-2105CCMG_3, 15 ml / kg, i.g. CDD-2105CCMG_4 CDD-2105CCMG_4, 15 ml / kg, i.g.

[0172] Experiment 2: Six- to eight-week-old male C57BL / 6J mice were used. Before the experiment, the baseline pain threshold of the mice was measured. Mice with pain thresholds ranging from 5 to 30 seconds were selected and randomly divided into five groups: a control group, a linaclotide group, a CDD-2105PCME (0.5×) group, a CDD-2105PCME (1×) group, and a CDD-2105PCME (2×) group, with 12 mice in each group. The content of each component of CDD-2105 was the same as described in Table 4. The treatments for each group are shown in Table 6 below.

[0173] Table 6

[0174] Group Dosing condition Control group 0.5% carboxymethyl cellulose sodium (CMC-Na), 15 ml / kg, i.g. Linaclotide 3.97 μg / ml linaclotide, 15 ml / kg, i.g. CDD-2105PCME (0.5x) CDD-2105PCME (0.5x), 15 ml / kg, i.g. CDD-2105PCME (1x) CDD-2105PCME (1x), 15 ml / kg, i.g. CDD-2105PCME (2x) CDD-2105PCME (2x), 15 ml / kg, i.g.

[0175] (2) Hot plate test method: On the day of the experiment, 3 hours after the mice were administered the drug by gavage, the mice were placed on a hot plate at 55±1℃. The time when the mice first licked their hind paws was recorded, and the pain threshold (latency of pain response) of the mice was measured. If the mice did not lick their paws within 30 seconds, the mice should be removed immediately to avoid burns. The pain threshold is calculated as 30 seconds.

[0176] (3) Results

[0177] like Figure 13 As shown, 3 hours after administration, the pain thresholds of mice in groups CDD-2105CCMG_2,_3, and_4 were significantly higher than those in the control group. In subsequent experiments, the content of the quality markers of traditional Chinese medicine in CDD-2105PCME(1×) was set to be comparable to that in CDD-2105CCMG_4, which had the lowest content of raw medicinal materials. Figure 14 As shown, 3 hours after administration, the pain threshold of mice in each CDD-2105PCME treatment group was significantly longer than that of the normal group, exhibiting a clear dose-response relationship. The effect of CDD-2105PCME (2×) was comparable to that of the clinically equivalent dose of linaclotide. These results indicate that CDD-2105 has a good alleviating effect on central pain perception impairment induced by heat stimulation in mice.

[0178] 2.3 Toxicity evaluation of the CDD-2105 mixture of traditional Chinese medicine quality markers

[0179] (1) Animals and drug administration

[0180] Six- to eight-week-old ICR mice, half male and half female, were randomly divided into a normal control group, and four groups: CDD-2105MM (10x), CDD-2105MM (20x), CDD-2105MM (50x), and CDD-2105MM (100x). Single-dose administration was performed using the up-down method. Mortality, body weight, and food intake changes were recorded after administration. On day 14 post-administration, the morphology, weight, and organ coefficients of the heart, liver, spleen, lung, and kidneys were observed. The traditional Chinese medicine quality markers of CDD-2105, including corydaline, magnolol, sennoside A, and paeoniflorin, were administered in combination. The total content of these quality markers in each combination is shown in Table 7 below.

[0181] Table 7

[0182] Composition Total content of traditional Chinese medicine quality markers (mg / kg) Clinical equivalent dose multiple CDD-2105MM (10x) 924.33 10x CDD-2105MM (20x) 1848.46 20x CDD-2105MM (50x) 4621.15 50x CDD-2105MM (100x) 9242.29 100x

[0183] (2) Statistical analysis

[0184] Statistical analysis was performed using GraphPad 10 software. Results are expressed as mean ± standard error. Differences between groups in defecation frequency, fecal water content, and pain response latency were analyzed using one-way ANOVA and Dunnett's multiple comparison test. Differences between groups in body weight and organ coefficients were analyzed using two-way ANOVA and Dunnett's multiple comparison test. A p-value < 0.05 was considered statistically significant.

[0185] (3) Results

[0186] Following a single administration of a high-dose mixture of CDD-2105 traditional Chinese medicine quality markers, no deaths occurred in either female or male mice during the observation period, and body weight ( Figure 15 ) and food intake ( Figure 16 No negative effects were observed. Fourteen days after administration, visual observation revealed no significant lesions in any organ, and the organ coefficient was not significantly affected by CDD-2105MM. Figure 17 The results showed that CDD-2105 had no serious acute toxicity risk and good safety.

[0187] In summary, the traditional Chinese medicine composition CDD-2105 shown in this invention can effectively relieve IBS-C symptoms without the risk of serious acute toxicity.

Claims

1. A composition of traditional Chinese medicine extract, wherein, The composition of the traditional Chinese medicine extract contains, based on the dry matter content of the composition of the traditional Chinese medicine extract, sennoside A 1-10%, sennoside B 1-10%, magnolol 1-10%, honokiol 1-10%, paeoniflorin 30-50%, albiflorin 5-10%, fumarine 0.1-0.5%, and stylopine 0.1-0.5%.

2. The composition of traditional Chinese medicine extract according to claim 1, wherein, The composition of the traditional Chinese medicine extract contains, based on the dry matter content of the composition of the traditional Chinese medicine extract, sennoside A 1-5%, sennoside B 1-5%, magnolol 1-5%, honokiol 1-5%, paeoniflorin 30-50%, albiflorin 5-10%, fumarine 0.1-0.3%, and stylopine 0.1-0.3%.

3. A composition of traditional Chinese medicine extract, wherein, The composition of the traditional Chinese medicine extract includes extracts of original medicinal materials; The original medicinal materials include Magnolia officinalis, Cassia obtusifolia, Paeonia lactiflora or Paeonia suffruticosa, Corydalis saxicola or Corydalis saxicola in vinegar; The mass ratio of the original medicinal materials Magnolia officinalis, Cassia obtusifolia, Paeonia lactiflora or Paeonia suffruticosa, Corydalis saxicola or Corydalis saxicola in vinegar is (1-20):(1-30):(1-25):(5-50); Preferably, the mass ratio of the original medicinal materials Magnolia officinalis, Cassia obtusifolia, Paeonia lactiflora or Paeonia suffruticosa, Corydalis saxicola or Corydalis saxicola in vinegar is (10-20):(1-10):(15-25):(5-15) or (1-2):(1-10):(1-10):(20-50).

4. A pharmaceutical composition for treating constipation, which is composed of the composition of the traditional Chinese medicine extract according to any one of claims 1-3 and one or more pharmaceutically acceptable excipients.

5. The pharmaceutical composition of claim 4, wherein, The dosage form of the pharmaceutical composition includes one or a combination of more than two of capsules, tablets, powders, solutions, or suspensions.

6. A method for preparing the composition of the traditional Chinese medicine extract according to any one of claims 1 to 3 or the pharmaceutical composition according to claim 4 or 5, wherein, The preparation method includes: The composition or the pharmaceutical composition is obtained by mixing Magnolia officinalis extract, Cassia obtusifolia extract, Paeonia lactiflora or Paeonia suffruticosa extract, and Corydalis saxicola or Corydalis saxicola in vinegar extract.

7. The preparation method according to claim 6, wherein: The preparation method of the Magnolia officinalis extract includes: extracting Magnolia officinalis with water or ethanol to obtain Magnolia officinalis extract liquid, further concentrating to obtain Magnolia officinalis extract concentrate, and extracting the Magnolia officinalis extract concentrate with petroleum ether to obtain Magnolia officinalis extract; The preparation method of the Cassia obtusifolia extract includes: extracting Cassia obtusifolia with water, sodium bicarbonate, or ethanol to obtain Cassia obtusifolia extract liquid, further concentrating to obtain Cassia obtusifolia concentrated liquid, and purifying the Cassia obtusifolia concentrated liquid by column to obtain Cassia obtusifolia extract; The preparation method of the Paeonia lactiflora or Paeonia suffruticosa extract includes: extracting Paeonia lactiflora or Paeonia suffruticosa with water or ethanol to obtain Paeonia lactiflora or Paeonia suffruticosa extract liquid, further concentrating to obtain Paeonia lactiflora or Paeonia suffruticosa concentrated liquid, and purifying the Paeonia lactiflora or Paeonia suffruticosa concentrated liquid by column to obtain Paeonia lactiflora or Paeonia suffruticosa extract; The preparation method of the Corydalis saxicola or Corydalis saxicola in vinegar extract includes: extracting Corydalis saxicola or Corydalis saxicola in vinegar with water or ethanol to obtain Corydalis saxicola or Corydalis saxicola in vinegar extract liquid, further concentrating to obtain Corydalis saxicola or Corydalis saxicola in vinegar concentrated liquid, and purifying the Corydalis saxicola or Corydalis saxicola in vinegar concentrated liquid by column to obtain Corydalis saxicola or Corydalis saxicola in vinegar extract.

8. The preparation method according to claim 6, wherein: In the preparation method of the Magnoliae officinalis extract, the temperature for water or ethanol extraction is 40-90 DEG C; the temperature for petroleum ether extraction is 40-70 DEG C; In the preparation method of the Folii sennae extract, the temperature for water, sodium bicarbonate or ethanol extraction is 25-90 DEG C; In the preparation method of the Radix paeoniae alba or Radix paeoniae rubra extract, the temperature for water or ethanol extraction is 60-90 DEG C; In the preparation method of the Rhizoma corydalis or Rhizoma corydalis vinegar extract, the temperature for water or ethanol extraction is 60-90 DEG C.

9. The production method according to claim 6, wherein The preparation method further comprises a step of detecting the active ingredients in the traditional Chinese medicine extract composition of any one of claims 1-3 or the pharmaceutical composition of claim 4 or 5; Preferably, the detection is liquid chromatography-tandem mass spectrometry detection.

10. Use of the traditional Chinese medicine extract composition of any one of claims 1-3, the pharmaceutical composition of claim 4 or 5 or the preparation method of any one of claims 6-9 in the preparation of a drug for preventing and / or treating constipation. Preferably, the constipation is abdominal pain type constipation.

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