Drug for treating urolithiasis-related diseases and preparation method thereof

Abstract

The present invention provides a medicinal active ingredient extract extracted from Polygala gracilis. The extract may contain a flavonol compound having the structure of the following formula (I) as a first active ingredient, a xanthone compound selected from the following formula (II) as a second active ingredient, and a glycolipid compound selected from the following formula (III-1) as a third active ingredient. Animal experiments have confirmed that the drug of the present invention has significantly superior effects in typical test indicators such as calcium oxalate crystal aggregation, renal interstitial inflammatory cell infiltration, and renal tubular dilatation lesions compared with potassium sodium hydrogen citrate, and have shown that the drug has high potential and market prospects as an adjuvant drug for the treatment of urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and after surgical treatment of urolithiasis. [C1] TIFF2024504686000070.tif40156[2] TIFF2024504686000071.tif29156[3] TIFF2024504686000072.tif56156

Description

[Technical field]

[0001] REFERENCE TO RELATED APPLICATIONS This application claims priority to Chinese Patent Application No. 202110060462.2, filed on January 18, 2021, the entire contents of which are incorporated herein by reference.

[0002] The present invention belongs to the field of medicinal chemistry. Specifically, the present invention provides a drug for treating urolithiasis, which is an extract of the plant Polygala gracilis, preferably an extract of the stem and leaf parts of Polygala gracilis, containing structurally specific flavonol compounds, xanthones, and glycolipids as the main active ingredients. The drug of the present invention is obviously superior to the existing mainstream clinical drug potassium sodium hydrogen citrate, and has fewer side effects in treating urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, or as an adjuvant drug after surgical treatment of urolithiasis, and therefore has high medical and economic value. [Background technology]

[0003] Urolithiasis (also called urinary stones) is the oldest disease discovered in humans. Urinary stones were discovered 6,800 years ago, in 4800 BC, at the ruins of El Amra in Egypt. More than 2,000 years ago, Hippocrates described renal abscesses caused by kidney stones and also described arthrolithiasis. Also, 2,000 years ago in China, there was a description of urinary stones and painful dysuria due to urinary stones. Urolithiasis is not only a human disease, but also occurs in animals.

[0004] Urolithiasis is a general term for diseases caused by urinary stones formed by precipitation and concentration of crystals in urine. The development of urolithiasis is associated with supersaturation of stone-forming salts (e.g., excessive excretion of salts, uric acidity, low urine output), preformed nuclei (e.g., uric acid crystals and other stones), and the presence in the urine of factors that promote crystallization formation inhibitor abnormalities. Idiopathic hypercalciuria [urinary calcium >300 mg / d (>7.5 mmol / d) in men and >250 mg / d (6.2 mmol / d) in women] is a genetic disorder that occurs in 50% of men with calcium stones and 75% of women with calcium stones. Hypocitraturia [urinary citrate <350 mg / d (<1820 μmol / d)] alone or in combination with other diseases may promote stone formation, since citrate normally binds urinary calcium to form soluble calcium citrate.

[0005] There are many areas in the world with a high incidence of urinary stones, called stone regions. This mainly refers to areas with a high incidence of urinary stones due to malnutrition in children. Areas with a relatively high incidence of urinary stones include the southeastern United States, the United Kingdom, the Nordic countries, the Mediterranean countries, northern India, Pakistan, northern Australia, Central Europe, the Malay Peninsula, and southern China. Areas with a low incidence include Central America, South America, and Africa. The average incidence of urinary stones in China is 5%, and there are about 50 million patients with urinary stones. This distribution has a certain regional distribution. The incidence is relatively high in areas such as Guangdong, Guangxi, Yunnan, Guizhou, Shandong, Hunan, Jiangxi, and Anhui. The incidence in the south is significantly higher than that in the north. The lowest incidence is 2.5% in Heilongjiang Province, and the highest incidence is 59% in Guizhou Province, a difference of 20 times. Furthermore, the recurrence rate of urolithiasis is very high, approximately 15-50%.

[0006] For a long time, people have been searching for an effective treatment method for urinary tract stones, including extracorporeal shock wave lithotripsy, flexible ureteroscopic lithotripsy, minimally invasive percutaneous nephrotripsy (stone diameter ≦2cm), open surgery (stone diameter ≧2cm), drug therapy (stone diameter <0.6cm), and other methods. Among these, extracorporeal shock wave lithotripsy for stones at the upper, middle, and lower tubular orifices will inevitably cause ureteral damage, and impaction of stones in the renal pelvis may also cause kidney damage, and clinically hematuria is evident. Lithotripsy and stone removal by flexible ureteroscope may damage the ureter and kidney, resulting in incomplete stone removal. Lithotripsy and stone removal by minimally invasive percutaneous nephrotripsy may lead to kidney damage and incomplete stone removal. With the development of clinical medicines, open surgery is rarely used to treat kidney stones.

[0007] Clinically, the treatment of urinary tract stones involves the use of excretory agents such as potassium sodium hydrogen citrate granules, potassium citrate, thiazide diuretics, magnesium, and acetylcysteine ​​as Western medicines, but their effects are not ideal and their toxicity and side effects are obvious. Potassium sodium hydrogen citrate granules (trade name: Uralyt-U) was developed in 1965 by MADAUS AG in Germany as the world's first citrate preparation to dissolve and prevent uric acid stones. In 2005, it was recommended by the Urolithology Group of the Urology Branch of the Chinese Medical Association as the only domestically legal citrate preparation with independent chemical structure and stone-dissolving effect. However, potassium sodium hydrogen citrate granules need to be taken in a very high effective dosage, with a daily dosage of 4 bags (each bag is 2.5 grams of granules, a total of 10 grams of granules), taken three times after meals: one bag in the morning, one bag at noon, and two bags at night. The granules can be taken with water. One gram of potassium sodium hydrogen citrate contains 0.172 grams or 4.4 mmol of potassium and 0.1 grams or 4.4 mmol of sodium (equivalent to 0.26 grams of sodium chloride). Daily intake of such large amounts of sodium and potassium ions can cause serious diseases such as severe hyperkalemia, cardiac arrhythmias, and hypertension, and therefore the scope of use of this drug is severely limited.

[0008] In China, a large number of herbal medicines and Chinese patent medicines have also been explored in clinical and medical research as treatment plans. For example, Chinese patent applications CN103285355A, CN103704591A, CN104083644A, CN105213919A, CN105998861A, CN1653929A, etc., describe a series of multi-component Chinese herbal medicines for treating urolithiasis. However, multi-component Chinese herbal medicines generally have problems such as complex ingredients, underdeveloped pharmaceutical technology, difficulty in quality control, inaccuracy of quantitative detection methods, large dosages, incomplete quality control standards, and inconsistency with modern clinical pharmaceutical standards and drug requirements. In addition, some traditional Chinese patent medicines, such as Bishitong, Paishi Granules, Shilintong Tablets, and Relinqing Granules, are also common Chinese patent medicines used to treat urolithiasis. However, these traditional Chinese patent medicines have problems such as complex ingredients, underdeveloped pharmaceutical technology, difficult quality control, inaccurate quantitative detection methods, large dosages, incomplete quality control standards, and inconsistency with modern clinical drug regulations.

[0009] With the continuous deepening of the reform of the Chinese drug evaluation system, the quality requirements of drugs declared for registration continue to increase. Among these, for the clinical trials of Chinese herbal medicines and their active extracts, the new approval standards generally require double-blind comparative trials compared with typical Western medicines (such as potassium sodium hydrogen citrate for the treatment of urolithiasis) for the same indicators, and only if the clinical efficacy and safety of the natural medicines or their extracts are the same as or better than those of Western medicines (compound medicines) with the same indicators of clinical trials, the sale of natural medicines or their extracts can be approved. Due to the high evaluation standard requirements of natural medicines, currently, 40-50 chemically and biologically novel drugs are approved for production every year in China, while only 1-2 are approved for production every year for traditional Chinese medicines or natural medicines. Therefore, although there are pharmaceutical companies that try to extract active active extracts and active medicinal ingredients from various Chinese herbal medicines for the treatment of urolithiasis in recent years, few of them have obtained production approval. Even individual drugs that have entered phase 2 / 3 clinical trials have ended in failure. One of the important reasons is that research on the active parts and medicinal ingredients of these drugs is insufficient, resulting in the clinical efficacy and safety of these drugs being significantly inferior to existing typical clinical Western medicines such as potassium sodium hydrogen citrate, and therefore they could not pass the evaluation tests and were not approved for manufacturing.

[0010] Therefore, developing a drug for urolithiasis that is safe and effective, has stable and controllable quality, obvious therapeutic effect, few side effects (same as or less than that of potassium sodium hydrogen citrate, the existing main drug for urolithiasis), is well absorbed by the body, and meets the modern drug registration requirements, at low cost and with simple process, is still an urgent pursuit of the medical community.

[0011] Polygala gracilis is a plant of the Polygalaceae family and is widely distributed in China, where it is widely used in traditional medicine, mainly for removing phlegm, relieving coughs, releasing blood congestion, stopping bleeding, calming the heart and nerves, detoxifying, and reducing swelling.

[0012] The functions and indications of the traditional Chinese medicine Polygala gracilis, as listed in the "Chinese Pharmacopoeia" (2015 edition, Vol. 1), are to clear phlegm, relieve cough, promote blood circulation, reduce swelling, detoxify, and relieve pain. It is also used for cough with excess phlegm, sore throat, external treatment of wounds, swelling, skin boils, and snakebites.

[0013] The preparation of Polygala gracilis listed in the "Chinese Pharmacopoeia" (2015 edition, Vol. 1) is Polygala gracilis granular compound. The formula is 150g Polygala gracilis, 350g Daqing Bao Bao Bao, 200g Wild Chrysanthemum, 250g Spora Lygodii, 250g Herba Hedyotis, 200g Herba Violae. Functions and Indications: Clearing heat and relieving sore throat, dissipating stagnation, relieving pain, clearing phlegm, and relieving cough. It is used for pharyngeal swelling, sore throat, fever, and cough caused by wind-heat attacking the lungs or phlegm-heat inhibiting the lungs; acute pharyngitis, acute exacerbation of chronic pharyngitis, and upper respiratory tract infections with the above symptoms.

[0014] There are also many patent documents that describe the extraction of various medicinal components of Polygala for use as medicines. For example, patent CN1303097C describes polygala saponins and their aglycones, total saponins and their total aglycones, and their effects in treating depression, developing intelligence, sedation, anxiolysis, and hypnosis. Patent CN104004110B describes the application of Polygala polysaccharides extracted from Polygala in the preparation of drugs and health foods for improving the immune function of the body. Patent CN108159126A describes the application of polygala saponin extract in the preparation of anti-cancer drugs. Patent CN103006793B describes the separation and purification process of the effective anti-inflammatory part of Polygala, and discloses that the total flavonoids and total saponin extract of Polygala are the anti-inflammatory effective parts. Patent CN108948125A describes a method for preparing Polygala sapogenin using Polygala gracilis, a method for preparing Polygala sapogenin, and a method for preparing Polygala flavones, the latter of which mentions four specific flavone molecules, kaempferol-3-O-6''-O-(3-hydroxy-3-methyl-glutaryl)glucoside, astrogrin, kaempferol 3-(6-acetyl)glucoside, and kaempferol 3,7-diglucoside. However, the pharmaceutical activity of the extracted components has not been confirmed. Also, existing published literature does not disclose the therapeutic use of specific extracts of Polygala gracilis for urolithiasis.

[0015] In addition, all of the above patents use the whole part of Polygala gracilis and use saponin compounds as essential medicinal ingredients. The quality control detection standard for Polygala gracilis compounds listed in the "Chinese Pharmacopoeia" (2015 edition, Vol. 1) is that the content of Polygala gracilis saponin (C53H86O23) calculated as a dry product should not be less than 0.60%. However, the chemical components of Polygala gracilis include saponins, flavonoids, glycolipids, alkaloids, phenols, tannins, polysaccharides, etc., and these components are very complex. According to a CNKI literature search, there are more than 100 compounds with confirmed molecular structures. Therefore, it is difficult to develop all parts of Polygala gracilis into natural medicines that meet modern pharmaceutical standards. In fact, only preparations of Polygala gracilis compounds are currently distributed, and Polygala gracilis used as a single herbal medicine is not distributed. Summary of the Invention

[0016] The main objective of the present invention is to provide a drug for urolithiasis, which is safe and effective, has stable and controllable quality, obvious therapeutic effect, few side effects (same as or less than that of potassium sodium hydrogen citrate, the existing main drug for urolithiasis), is well absorbed by the body, and meets the modern drug registration requirements, at low cost and with a simple process.

[0017] Through extensive experimental studies, the present inventors have discovered a medicinal active ingredient extract of a flavonol compound having a specific structure of general formula (I) extracted from the plant Polygala gracilis.

[0018] Specifically, the present invention provides a medicinal active ingredient extract extracted from the plant Polygala gracilis, the extract containing a flavonol compound of the following formula (I) as a first active ingredient: [ka] wherein R1 is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Api, -ORha, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Api, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Api, -O-Gal-Rha, -O-Glc-Glc-Api, -O-Gal-Glc-Api, -O-Glc-Glc-Api, -O-Gal-Gal-Api, -O-Gal-Gal-Api, -O-Gal-Rha-Gal, -O-Gal-Rha-Gal, -O-Gal-Rha-Glc, -O-Glc-Rha-Glc, and -O-Glc-Rha-Gal; R2 is a substituent selected from the group consisting of -OH, -O-Me, -O-Glc, -O-Gal, -O-Api, and -O-Rha; R3 is a substituent selected from the group consisting of H, OH, -O-Me, -O-Glc, -O-Gal, -O-Api, and -O-Rha; R4 is a substituent selected from the group consisting of OH and -O-Me.

[0019] The medicinal active ingredient extract may contain a xanthone compound selected from the following formula (II) as a second active ingredient, and a glycolipid compound selected from the following formula (III) as a third active ingredient. [ka] Here, R5 is a substituent selected from the group consisting of -O-Gal, -O-Api, -ORha, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Api, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Api, -O-Gal-Rha, -O-Api-Glc, -O-Api-Gal, -O-Api-Api, and -O-Api-Rha, and R6 is a substituent selected from the group consisting of -OH and -O-Me. [ka] wherein R7, R8 are each independently selected from the group consisting of H and CH3; In the definitions of R1 to R6 in the above general formula (I) and general formula (II), Glc represents glucosyl, Gal represents galactosyl, Api represents celosyl, and Rha represents rhamnose.

[0020] In a preferred technical solution, the flavonol compound of general formula (I) as the first active ingredient is preferably selected from one or more compounds represented by the following general formula: [ka] wherein R1 is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Rha, -O-Glc-Glc-Api, -O-Gal-Glc-Api, -O-Glc-Gal-Api, and -O-Gal-Gal-Api. [ka] wherein R1 is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Api, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Api, -O-Gal-Rha, -O-Glc-Glc-Api, -O-Gal-Glc-Api, -O-Glc-Gal-Api, -O-Gal-Gal-Api, -O-Gal-Rha-Gal, -O-Gal-Rha-Gal, -O-Gal-Rha-Glc, -O-Glc-Rha-Glc, and -O-Glc-Rha-Gal. [ka] wherein R1 is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Api, and -O-Gal-Api. [ka] wherein R is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Rha, and -O-Gal-Rha. [ka] wherein R is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Rha, and -O-Gal-Rha. [ka] wherein R is selected from the group consisting of -OH, -O-Gal, and -O-Gal-Api.

[0021] The xanthone compound of general formula (II) as the second active ingredient is preferably selected from one or more of the following formulae (II-1), (III-2), and (II-3). [ka] [ka] [ka]

[0022] The glycolipid compound as the third medicinal ingredient is preferably a compound of the following formula (III-1) or (III-2). [ka] [ka]

[0023] In a further preferred technical solution, the first active ingredient in the pharmaceutical active ingredient extract of Polygala gracilis is selected from the flavonol compounds of the above general formulas F-7K, F-7Q, F-74Q, and F-74K.

[0024] In a further preferred technical solution, the first active ingredient in the medicinal active ingredient extract of Polygala gracilis is preferably at least one selected from the following compounds: [ka] [ka] [ka]

[0025] In a further preferred technical solution, the total content of the flavonol compound of general formula (I) as the first medicinal active ingredient and the xanthone of general formula (II) and glycolipid of general formula (III) as the second medicinal active ingredient, which may be contained, in the medicinal active ingredient extract of Polygala gracilis is 30-100% of the total extract of Polygala gracilis, where the component content (%) is the HPLC% content calculated using the HPLC integral area normalization method according to a method commonly used in the art.

[0026] In a more preferred technical solution, the total content of the flavonol compound of general formula (I) as the first medicinal ingredient in the medicinal active ingredient extract of Polygala gracilis is 20-100% of the total extract of Polygala gracilis.

[0027] In a more preferred technical solution, the total content of the flavonol compound of general formula (I) as the first medicinal ingredient in the medicinal active ingredient extract of Polygala gracilis is 75-100% of the total extract of Polygala gracilis.

[0028] The present invention also provides a method for preparing a pharmaceutical active ingredient extract of Polygala gracilis, said method comprising the steps of: (1) Pretreatment of Polygala The whole or above-ground parts of Polygala gracilis, or commercially available medicinal materials of Polygala gracilis, are taken, washed and crushed to obtain raw materials of Polygala gracilis. (2) Rough extraction of the effective parts of Polygala gracilis A portion of the raw material Polygala obtained in step (1) is taken and heated and refluxed for 1 to 3 hours each time with ethanol at a concentration of 20 to 95% (v / v), which is approximately 6 to 12 times the weight of the raw material Polygala, and extracted 1 to 3 times by repeatedly refluxing. The obtained alcohol extracts are combined and concentrated by filtration or centrifugation to obtain a total alcohol extract of Polygala. or A portion of the raw material Polygala obtained in step (1) is heated and boiled in deionized water in an amount approximately 6 to 15 times the weight of the Polygala. The boiling is continued for 1 to 3 hours, and extraction is repeated 1 to 3 times. The resulting aqueous extracts are combined, filtered or centrifuged, and concentrated to obtain a total aqueous extract of Polygala. (3) Purification of the active parts of Polygala gracilis The concentrate of the total aqueous extract or total alcohol extract of Polygala gracilis in step (2) is separated using a macroporous adsorption resin or polyamide resin chromatographic column, and gradient elution is performed successively with different ratios of water / ethanol until the eluate becomes colorless. The 0-95% ethanol gradient eluate is collected and dried under reduced pressure to obtain the desired extract of medicinal active ingredients of Polygala gracilis.

[0029] In the above extraction method, in step (3), the macroporous resin is preferably selected from macroporous resins of type D101, HPD100, HPD200, or AB-8, and the polyamide resin is preferably selected from SCR polyamide resins of 100-200 mesh.

[0030] In the above extraction method, the Polygala gracilis in step (1) is preferably the stem and leaf parts of Polygala gracilis.

[0031] In the above extraction method, in step (3), the macroporous resin is preferably selected from macroporous resins of type D101 or AB-8, and the polyamide resin is preferably selected from SCR polyamide resins of 100-200 mesh.

[0032] In the above extraction method, it is preferred that the gradient elution is carried out in step (3) by successively eluting with water, 25% ethanol, 50% ethanol, 75% ethanol, and 95% ethanol until the eluate becomes colorless.

[0033] The present invention provides the use of a medicinal active ingredient extract of Polygala gracilis in the preparation of a medicament for treating or preventing urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug after surgical treatment of urolithiasis.

[0034] The present invention also provides a pharmaceutical composition comprising, as an active ingredient, at least one compound selected from the following formulae: [ka] [ka] [ka]

[0035] In a preferred technical solution, said pharmaceutical composition further comprises a pharma- ceutically acceptable carrier, excipient, or auxiliary material.

[0036] The present invention further provides the use of the above pharmaceutical composition in the preparation of a medicament for the treatment or prevention of urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug following surgical treatment of urolithiasis.

[0037] The invention also provides the use of any one of the following compounds in the preparation of a medicament: [ka] [ka] [ka]

[0038] The medicament is used for the treatment or prevention of urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug after surgical treatment of urolithiasis.

[0039] Through many experimental studies, the inventors have found that the medicinal ingredient extract of the present invention can be used as an adjuvant drug for the treatment of urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and after surgical treatment of urolithiasis, and is significantly more effective than the herbal medicines and crude drug extracts currently known in published literature, and has the same or better effect as the well-known clinical Western medicine potassium sodium hydrogen citrate (see the pharmacological examples below for details). Preliminary studies have shown that it is important that the compounds of general formula (I) and general formula (II) selected as the main medicinal ingredients of the present invention have a hydroxyl group as a substituent at the β-position of the carbonyl of flavonoids and xanthones. Here, the hydroxyl group and the ketone carbonyl group act together to more effectively react with the calcium ion-containing components of urinary stones, thereby more effectively decomposing or dissolving urinary stones.

[0040] Therefore, a specific technical solution of the present invention provides the use of any one of the compounds of general formula (I) and general formula (II) or a combination of two or more of them in the preparation of a medicament for treating or preventing urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug after surgical treatment of urolithiasis.

[0041] In a preferred technical solution of the present invention, there is provided the use of any one of the compounds of the above general formulae (I-1)-(I-4) and / or general formulae (II-1)-(II-3), or a combination of two or more of them, in the preparation of a medicament for treating or preventing urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug after surgical treatment of urolithiasis.

[0042] The pharmaceutical active ingredient extract of Polygala gracilis of the present invention can also be prepared into various dosage forms, such as enteral dosage forms such as capsules, tablets, pills, oral liquids, granules, tinctures, sustained release preparations, and parenteral dosage forms such as injections and topical preparations, by conventional pharmaceutical preparation methods. [Brief description of the drawings]

[0043] [Figure 1] FIG. 1 is a HPLC analysis chromatogram of the alcohol extract from all parts of Polygala gracilis of the present invention. [Diagram 2] FIG. 2 is a HPLC analysis chromatogram of the alcohol extract from the aerial parts of Polygala gracilis of the present invention. [Diagram 3] FIG. 3 is an HPLC analysis chromatogram of the effective portion obtained after gradient elution of the aerial parts of Polygala gracilis of the present invention with polyamide resin. [Figure 4] FIG. 4 is a CH correlation two-dimensional NMR chromatogram of the compound F-7Q-1 of the present invention. [Diagram 5] FIG. 5 is a CH remote two-dimensional NMR correlation chromatogram of the compound F-7Q-1 of the present invention. [Figure 6] FIG. 6 is a CH correlation two-dimensional NMR chromatogram of the compound F-7K-1 of the present invention. [Figure 7] FIG. 7 is a CH remote correlation two-dimensional NMR chromatogram of the compound F-7K-1 of the present invention. [Figure 8] FIG. 8 is a CH correlation two-dimensional NMR chromatogram of the compound F-74Q-1 of the present invention. [Figure 9] FIG. 9 is a CH remote correlation two-dimensional NMR chromatogram of the compound F-74Q-1 of the present invention. [Figure 10] FIG. 10 shows the results of HE microscopic observation of renal tubular dilatation lesions in an animal experiment using the drug of the present invention (normal group). [Figure 11]FIG. 11 shows the results of HE microscopic observation of renal tubular dilatation lesions in an animal experiment using the drug of the present invention (model group). [Figure 12] FIG. 12 shows the results of HE microscopic observation of renal tubular dilatation lesions in an animal experiment using the drug of the present invention (potassium sodium hydrogen citrate drug group). [Figure 13] FIG. 13 shows the results of HE microscopic observation of renal tubular dilatation lesions in an animal experiment using the drug of the present invention (low dose group). [Figure 14] FIG. 14 shows the results of HE microscopic observation of renal tubular dilatation lesions in an animal experiment using the drug of the present invention (medium dose group). [Figure 15] FIG. 15 shows the results of HE microscopic observation of renal tubular dilatation lesions in an animal experiment using the drug of the present invention (high dose group). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0044] In the following, the embodiments of the present invention will be described in detail. The diagrams of the embodiments are shown in the attached drawings. The embodiments described below with reference to the drawings are merely exemplary and are intended to explain the present invention, and should not be understood as limiting the present invention.

[0045] I. Preparation example Preparation Example 1: Component analysis of alcohol extract from all parts of Polygala gracilis All the pieces of Polygala were weighed and 10 times the amount of 75% ethanol was added. The resulting solution was heated to reflux and extracted three times for 3 hours each time, filtered hot, and then the alcohol extracts were combined.

[0046] The alcoholic extract was concentrated to obtain an extract concentrate with a relative density of 1.1-1.3 g / ml.

[0047] The fingerprint analysis of the obtained alcohol extract concentrate of Polygala gracilis was carried out using HPLC. HPLC test conditions: Mobile phase: acetonitrile (A), 0.1% formic acid in water (B), binary gradient separation Flow rate: 0.8mL min -1 Detection wavelength: 330nm Column temperature: 20℃ Injection volume: 20μL Chromatograms were recorded for 90 min.

[0048] Preparation Example 2: Component analysis of water extract from all parts of Polygala gracilis All the pieces of Polygala were weighed and 10 times the amount of deionized water was added. The resulting solution was heated to reflux and extracted three times for 3 hours each time, filtered hot, and then the water extracts were combined. The extract is a dark brown liquid.

[0049] The aqueous extract was concentrated to obtain an extract concentrate with a relative density of 1.1-1.3 g / ml.

[0050] The fingerprint analysis of the obtained Polygala water extract concentrate was carried out using HPLC (see Figure 1 in the attached drawings for details). It can be basically confirmed that the extract mainly contains four major compounds and several dozens of other compounds. In the HPLC chromatogram, the peak time of xanthones is between 12 and 25 minutes, the peak time of flavonol compounds is between 18 and 68 minutes, the peak time of glycolipids is between 15 and 45 minutes, and the peak time of saponins is between 42 and 85 minutes.

[0051] Comparing the components obtained by the aqueous extraction method with those obtained by the alcohol extraction method, it can be seen that the extract obtained by the aqueous extraction method contains less impurities with low polarity (such as chlorophyll) and more highly polar tannins due to the relatively high polarity of water. The extract is dark brownish yellow in color. However, the alcohol extract contains more impurities with low polarity (such as chlorophyll) and significantly less impurities with high polarity (such as tannins). The alcohol extract is greenish in color, but turns brownish yellow after cooling.

[0052] Extraction of the whole herb consumes a lot of medicinal plants. Considering that most of the commercially available Polygala gracilis is the aerial part, it is also beneficial to protect medicinal plant resources by selecting the aerial part and only keeping the roots of the plant. Therefore, the inventor further tried to extract medicinal ingredients from the aerial part of Polygala gracilis by alcohol extraction. The specific method is as follows:

[0053] Preparation Example 3: Component analysis of the water extract from the aerial parts of Polygala gracilis The aerial parts of Polygala gracilis were weighed and deionized water was added in an amount 10 times that of the medicinal material. The resulting solution was heated to reflux and extracted three times for three hours each time, filtered hot, and then the water extracts were combined.

[0054] The aqueous extract was concentrated to obtain an extract concentrate with a relative density of 1.1-1.3 g / ml.

[0055] A fingerprint analysis of the concentrate of deionized water extract of the aerial parts of Polygala gracilis was carried out using HPLC.

[0056] Preparation Example 4: Component analysis of alcohol extract from the aerial parts of Polygala gracilis The aerial parts of Polygala gracilis were weighed and 50% ethanol was added in an amount 10 times that of the medicinal material. The resulting solution was heated to reflux and extracted three times for three hours each time, filtered while hot, and then the alcohol extracts were combined.

[0057] The alcoholic extract was concentrated to obtain an extract concentrate with a relative density of 1.1-1.3 g / ml.

[0058] The fingerprint analysis of the obtained alcohol extract concentrate of the aerial parts of Polygala gracilis was carried out using HPLC (see FIG. 2 for the results).

[0059] The extract concentrates were analyzed by HPLC-MS to confirm their chemical composition. MS analysis included positive and negative ion analysis, MS-MS analysis, and MS-MS-MS analysis. The analytical results were compared with existing literature, and the structural types of four major categories were preliminarily confirmed: flavonol compounds, xanthones, glycolipids, and saponins. Furthermore, the flavonol compounds were further divided according to the differences in the parent nucleus of the flavonol compounds.

[0060] However, due to the complexity and diversity of the spatial structure and glycosyl binding methods in glycosides, it is possible to determine whether the binding of multiple glycosides to the parent nucleus of a flavonol compound is a single glycan, and the order of glycoside fragmentation in mass spectrometry by HPLC and multi-stage tandem MS to confirm the molecular weight of the terminal sugar of the glycan and the basic type of glycogen. However, due to the complexity and diversity of the spatial structure and glycosyl binding methods in glycosides, it is necessary to further identify by other means the arrangement of hydroxyl groups on specific carbon atoms of isomeric sugar rings (e.g., glucose or galactose), the position of the glycoside-glycosidic bond (e.g., 1-2 bond, 1-4 bond, or 1-6 bond), and the α- or β-configuration of the sugar configuration. Therefore, only by the structure identified by HPLC-MS-MS, a compound with the same molecular formula may have multiple structural combinations of the aglycon of a flavonol compound and different types of glycosides.

[0061] To facilitate the distinction, the present application uses compound categories to identify and distinguish compounds that may have various combinations of building blocks. For example, in the present application, F is used to represent a class of flavonol compounds, and FQ is used to represent a class of flavonol glycosides, where the parent nucleus of the flavonol compounds is quercetin. In determining the structure, 302-162-132 represents that the parent nucleus of the flavonol aglycon is quercetin, to which glucose (or galactose) is bound, and to which cellulose is bound. Here, 302 is the parent nucleus of quercetin, 162 is the molecular weight of the characteristic peak of the glucose (or galactose) fragmentation fragment in the mass spectrum, and 132 is the molecular weight of the characteristic peak of the cellulose fragmentation fragment.

[0062] It has been previously confirmed by HPLC and multi-stage tandem MS analysis that the alcohol extract from the aerial parts of Polygala gracilis contains mainly the following components:

[0063] [Table 1-1] [Table 1-2]

[0064] In the above table, the chemical composition structure mainly includes the following categories:

[0065] (1) Flavonols with glycosides 1. Flavonol glycoside compounds in the compound category F-7K. The aglycone structure of this compound is rhamnocitrin, 3,4',5-trihydroxy-7-methoxyflavone, or 7-methoxyl-kaempferol with a molecular weight of 300, and this compound has the following general structure: [ka] wherein R1=glycosyl and may be selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Rha, -O-Glc-Glc-Api, -O-Gal-Glc-Api, -O-Glc-Gal-Api, and -O-Gal-Gal-Api.

[0066] 2. Flavonol glycoside compound of compound category F-7Q. The aglycone structure of this compound is rhamnetin, 3,3',4',5-tetrahydroxy-7-methoxyflavone, or 7-methoxyl-quercetin with a molecular weight of 316, and this compound has the following general structure: [ka] wherein R1=glycosyl and may be selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Api, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Api, -O-Gal-Rha, -O-Glc-Glc-Api, -O-Gal-Glc-Api, -O-Glc-Gal-Api, -O-Gal-Gal-Api, -O-Gal-Rha-Gal, -O-Gal-Rha-Gal, -O-Glc-Rha-Glc, -O-Glc-Rha-Glc, and -O-Glc-Rha-Gal.

[0067] 3. Flavonol glycoside compound of compound category F-74Q. The aglycon structure of this compound is Ombuine with a molecular weight of 330, 3,5,3'-trihydroxy 7,4'-dimethoxy flavone, 7,4'-dimethoxyl-quercetin, and this compound has the following general structure: [ka] where R1=glycosyl and may be selected from the group consisting of OH, -O-Glc, -O-Gal, -O-Glc-Api, and -O-Gal-Api.

[0068] 4. Flavonol glycoside compound, whose compound category is FK. The aglycone structure of this compound is kaempferol or 3,4',5,7-tetrahydroxyflavone with a molecular weight of 286, and this compound has the following general structure: [ka] wherein R may be selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Rha, and -O-Gal-Rha.

[0069] 5. Flavonol glycoside compound, which is a compound category FQ. The aglycone structure is quercetin or 3,3',4',5,7-pentahydroxyflavone with a molecular weight of 302, and this compound has the following general structure: [ka] wherein R may be selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Rha, and -O-Gal-Rha.

[0070] 6. Flavonol glycoside compound of the compound category F-74K. The aglycone structure is ermanin, 3,5-dihydroxy 7,4'-dimethoxy flavone, or 7,4'-dimethoxyl-kaempferol with a molecular weight of 314, and this compound has the following general structure: [ka] where R=glycosyl and may be selected from the group consisting of -OH, -O-Gal, and -O-Gal-Api.

[0071] (2) Xanthone compounds in Polygala gracilis By secondary mass spectrometry, it can be confirmed that the xanthone compounds of Polygala gracilis are selected from the following formulae (II-1), (II-2), and (II-3). [ka] [ka] [ka]

[0072] (3) Glycolipid compounds By secondary mass spectrometry, it can be confirmed that the structure of the glycolipid compound is selected from the following formulas (III-1) and (III-2). [ka] [ka]

[0073] (4) Saponin compounds By HPLC and multi-stage tandem MS analysis, it can be confirmed that the alcohol extract from the aerial parts of Polygala gracilis of the present application contains polygalasaponin VIII, polygalasaponin XXI, polygalasaponin X, polygalasaponin XXIX, and other saponin compounds.

[0074] The alcohol extract from Polygala gracilis aerial part contains not only flavonols, xanthones and other target active ingredients, but also saponins, glycolipids and other ingredients. Therefore, the present inventors further refine the above alcohol extract from Polygala gracilis aerial part by separation methods such as macroporous resin and polyamide resin to separate and enrich the target active ingredients.

[0075] Preparation Example 5: Purification of alcohol extract from the aerial parts of Polygala gracilis with macroporous resin The extract concentrate obtained in Preparation Example 1 was passed through a D101 macroporous adsorption resin column at a flow rate of 1 column bed volume per hour. After the adsorption was completed, the column was first washed with 8 times the amount of resin to remove impurities, then washed with 0%-25, 25%-50, 50%-75, 75%-95% ethanol gradient elution solution of 2-5 times the column bed volume, and elution was performed at a flow rate of 0.5-2 times the column bed volume per hour to obtain the eluate. The ethanol elution solutions of different concentrations were concentrated 5-20 times, respectively, to obtain eluate concentrates with relative densities of 1.1-1.3 g / ml.

[0076] The components of the obtained alcohol extract concentrate of all parts of Polygala gracilis and the ethanol gradient eluate concentrate passed through a macroporous adsorption resin column using HPLC were subjected to fingerprint analysis.

[0077] Preparation Example 6: Purification treatment of the water extract from the aerial parts of Polygala gracilis with polyamide resin The extract concentrate obtained in Preparation Example 3 was passed through a polyamide resin column at a flow rate of 0.5 to 1 times the column bed volume per hour. After the adsorption was completed, the column was first washed with 2 to 8 times the amount of resin to remove impurities, and then washed with 0 to 25%, 25% to 50%, 50% to 75%, and 75% to 95% ethanol gradient elution solutions of 2 to 5 times the column bed volume, and elution was performed at a flow rate of 0.5 to 2 times the column bed volume per hour to obtain eluates. The above ethanol eluates of different concentrations were each concentrated 5 to 20 times to obtain eluate concentrates with relative densities of 1.1 to 1.3 g / ml.

[0078] Comparing the separation effects of the macroporous resin and the polyamide resin, it was found that the polyamide resin was able to remove more of the saponin components from the Polygala gracilis extract, and thus had a higher separation and purification effect.

[0079] The components of the extract concentrate of the aerial parts of Polygala gracilis obtained and the concentrate of the ethanol gradient eluate passed through a polyamide resin column using HPLC were subjected to fingerprint analysis.

[0080] HPLC analysis confirmed that the total content of the active ingredients (I), (II), and (III) was 50% to 90% of the orange to red eluate concentrate from the 0 to 25% ethanol elution portion (as shown in FIG. 3 herein).

[0081] The concentrate of the eluate was dried under reduced pressure at 75°C and crushed to obtain enriched medicinal ingredients of the aerial parts of Polygala gracilis, which were used in the drug efficacy comparison experiment.

[0082] The enriched active ingredients in the aerial parts of Polygala gracilis were analyzed by HPLC-MS. MS analysis included positive and negative ion analysis, MS-MS analysis, and MS-MS-MS analysis. The analytical results were compared with existing literature, and the compound structures shown in Table 2 below were confirmed.

[0083] [Table 2-1] [Table 2-2]

[0084] Based on the above analysis, it can be confirmed that the main components of Polygala extract, which has been extracted with water and purified with polyamide resin, include flavonol compounds of compound categories F-7K, F-7Q, F-74Q, and F-74K, xanthone compounds of general formula (II-1) (polygalaxanthone III), and glycolipid compounds.

[0085] In Table 2, the compound with a content of 18.97% was identified as F-7K-1, the compound with a content of 33.71% was identified as F-7Q-1, the compound with a content of 23.60% was identified as F-74Q-1, and the compound with a content of 4.81% was identified as compound F-74K-1 (Polygalitol B).

[0086] The meanings of the above compound category names are exactly the same as those in Table 1.

[0087] It should be emphasized that the degree of difference in the structure and content of the medicinal ingredients in the obtained medicinal ingredient extract may vary depending on the specific part of the plant Polygala parvum extracted (whole part, rhizome, or stem and leaves), the source, the preparation of the plant Polygala parvum (commercially available dried product, fresh Polygala parvum), as well as the conditions of the specific extraction and purification steps. In the present invention, the medicinal ingredient extract is preferably obtained from the fresh or dried aerial parts of Polygala parvum by water extraction and alcohol / water gradient elution on a polyamide column.

[0088] Preparation Example 7: Isolation and purification of the main active compounds Once the parent nucleus of a flavonol compound and the presence of multiple sugar rings in its molecule have been determined, it can be determined by HPLC-MS-MS whether it is a single sugar chain or not by obvious characteristics. However, there is insufficient basis to distinguish the arrangement of hydroxyl groups on a specific carbon atom of a sugar ring of the same molecular weight (e.g., glucose or galactose). When a sugar chain has more than two sugar rings, there is insufficient basis to determine the sugar bond position (e.g., 1-2 bond, 1-4 bond, or 1-6 bond) and the α- or β-configuration of the sugar structure. For this reason, the pure compounds of the main active ingredients were separated by semi-preparative HPLC and their specific structures were determined by a combination of H NMR, C NMR, and two-dimensional NMR.

[0089] The extract prepared in Example 5 was further separated and purified to obtain the pure single compounds of the three components with the highest content in the extract. That is, the pure single compounds of compounds F-7Q-1, F-7K-1, and F-74Q-1 were obtained.

[0090] The semi-preparative HPLC apparatus and conditions were as follows. HPLC test conditions Mobile phase: acetonitrile (A), deionized water (B), binary gradient separation Semi-preparative column: 19 x 250 mm, C18 Flow rate: 8mL min -1 Detection wavelength: 330nm Column temperature: room temperature Injection volume: 0.5mL The chromatogram was recorded for 180 min.

[0091] The structures of compounds F-7Q-1, F-7K-1, and F-74Q-1 were analyzed by NMR and two-dimensional NMR, respectively. The analytical results were as follows.

[0092] 1. Confirmation of the structure of compound F-7Q-1 Molecular weight: 640, sugar chain: 316-162-162

[0093] Important information regarding the 2D NMR of compound F-7Q-1 is summarized in the table below.

[0094] [Table 3]

[0095] Based on the above chromatographic analysis data, the exact spatial structure of the main active ingredient compound F-7Q-1 was finally confirmed to be as follows: [ka]

[0096] The chemical name of F-7Q-1 is rhamnetin 3-O-β-D-glucopyranosyl(1→2)-β-D-galactopyranoside or rhamnetin-3-O-(2″-O-β-D-glucopyranosyl)-β-D-galactopyranoside.

[0097] (2) Confirmation of the structure of compound F-7K-1 Molecular weight: 624, sugar chain: 300-162-162

[0098] Important information regarding the 2D NMR of compound F-7K-1 is summarized in the table below.

[0099] [Table 4]

[0100] Based on the above chromatographic analysis data, the exact spatial structure of the main active ingredient compound F-7K-1 was finally confirmed to be as follows: [ka]

[0101] The chemical name of F-7K-1 is rhamnocitrin 3-O-β-D-glucopyranosyl(1→2)-β-D-galactopyranoside, or OR rhamnocitrin-3-O-(2″-O-β-D-glucopyranosyl)-β-D-galactopyranoside.

[0102] 3. Confirmation of the structure of compound F-74Q-1 Molecular weight: 624, glycan: 330-162-132, identified

[0103] Important information regarding the 2D NMR of compound F-74Q-1 is summarized in the table below.

[0104] [Table 5]

[0105] Based on the above chromatographic analysis data, the exact spatial structure of the main active ingredient compound F-74Q-1 was finally confirmed to be as follows: [ka]

[0106] The chemical name of F-74Q-1 is 3,5,3'-trihydroxy-7,4'-dimethoxyflavone-3-O-β-D-apiofuranosyl(1→2)-β-D-galactopyranoside, polygallin C, or polygalitol C.

[0107] Based on the research of the present inventors, the inventors believe that the compounds F-7Q-1, F-7K-1, and F-74Q-1 isolated as the main components of the Polygala gracilis extract of the present invention play an important role in achieving the desired pharmaceutical effect. These compounds have a hydroxyl group as a substituent at the β-position of the carbonyl in the described molecular structure, and this hydroxyl group and the ketone carbonyl group act together to more effectively react with the calcium ion-containing components of stones in the urinary system, thereby more effectively decomposing or dissolving stones in the urinary system. Therefore, at least one of the compounds F-7Q-1, F-7K-1, and F-74Q-1 can also be used as the main and necessary active ingredients in the corresponding pharmaceutical composition for treating or preventing urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug after surgical treatment of urolithiasis. The pharmaceutical composition may further include a pharmaceutically acceptable adjuvant, carrier, or excipient.

[0108] II. Experimental examples of pharmacological activity 1. Preparation of Test Samples

[0109] [1] High-dose group test sample: The water extract prepared in Preparation Example 5, 0-25% ethanol gradient elution through a polyamide column, was concentrated and dried to obtain an extract of the active ingredient. A solution with a density of approximately 1.2 g / ml was prepared. The concentration of the active ingredient measured according to the pharmacopoeia standard (a standard curve was created using rutin as a standard substance) was approximately 130.4 mg / ml. [2] Medium dose group test sample: The high dose group sample was diluted once to obtain the medium dose group test sample. [3] Low dose group test sample: The medium dose group sample was diluted once to obtain the low dose group test sample. [4] Positive control test sample: Potassium sodium hydrogen citrate aqueous solution with a concentration of 100 mg / ml, 3 ml per day, equivalent to 300 mg / day.

[0110] 2. Animal Experiments 2.1. Experimental animals and feeding conditions Thirty-six SD rats were purchased from Shanghai Slack. License number: SCXK(Shanghai)2017-0005, certificate number: 20170005011248. Drinking water was ultrapure water. The license number of the experimental animal room was SYXK(Zhejiang)2015-0008.

[0111] Feeding environment: Temperature ranged from 20°C to 25°C, and relative humidity ranged from 40% to 70%. Adaptation feeding was performed for one week prior to the experiment.

[0112] 2.2. Experimental scheme 2.2.1 Experimental animals 36 SPF-grade male SD rats, 6-8 weeks old, 200-250 g

[0113] 2.2.3 Creating the model SD rats were fed an adaptive diet for 7 days, and all groups (except the normal group) were administered 1% ethylene glycol (in drinking water) + 2% ammonium chloride (intraenterically) at 2 ml / rat to establish the model for 28 consecutive days.

[0114] 2.2.4 Formation and treatment of experimental groups Thirty-six male SD rats were randomly divided into six groups (six rats in each group) according to body weight: normal group, model group, potassium sodium hydrogen citrate group, and traditional Chinese medicine extract (low, medium, high) dose group. During the modeling process, 3 ml of drugs were administered intraenterally every day. The drugs included positive control drugs and purified fractions passed through polyamide columns. After 4 weeks, the animals were euthanized.

[0115] 2.2.5 Kidneys The kidney tissue was peeled off in vivo, and one kidney was stored in a cryopreservation tube at -80°C to detect the Ca2+ concentration as tissue homogenate. The kit operation steps were the same as above. The other kidney was fixed in formalin solution, tissue sections were prepared, and HE staining was performed.

[0116] (1) The process for preparing paraffin sections was as follows. [1] Fixation, [2] Cutting, [3] Dehydration, [4] Clearing, [5] Immersion in wax, [6] Embedding, [7] Slicing, [8] Bake the slices, [9] Storage: Store in a box at room temperature.

[0117] (2) The HE staining process was as follows.

[0118] [1] Dewaxing and rehydration, [2] Dyeing, [3] Dehydration, penetration, and placement

[0119] [4] Staining results: the nuclei were blue, the cytoplasm was pink, and the red blood cells were bright red.

[0120] 3. Test Results 3.1. Positive test for calcium oxalate crystals in rat urine The results of urine routine report showed that rats in all other groups except normal group were positive for calcium oxalate crystals in urine.

[0121] 3.2. Serum Ca2+ concentration test results The difference in serum Ca2+ concentration between groups was small. Compared with the normal group, the model group and the medium dose treatment group showed significant differences. The difference in urinary Ca2+ concentration was large. Compared with the normal group, the low dose treatment group and the high dose treatment group showed highly significant differences (P<0.01). Compared with the model group, the low dose treatment group showed highly significant differences (P<0.01). The potassium sodium hydrogen citrate drug group had the highest concentration in kidney tissue, with highly significant differences compared with the model group and the normal group (P<0.01). The other groups did not show significant differences.

[0122] The amount of CRE in serum was, from highest to lowest, the model group > medium-dose treatment group > potassium sodium hydrogen citrate drug group > low-dose treatment group > high-dose treatment group > normal group. Compared with the normal group, the medium-dose treatment group and the high-dose treatment group showed significant differences (P<0.05), but there were no significant differences in the other groups.

[0123] The serum BUN level, from highest to lowest, was: model group > low-dose treatment group > potassium sodium hydrogen citrate drug group > medium-dose treatment group > high-dose treatment group > normal group. Compared with the normal group, all groups showed highly significant differences (P<0.01).

[0124] Compared with the model group, all the other groups showed highly significant differences (P<0.01), except for the low-dose treatment group, which showed significant differences (P<0.05).

[0125] 3.3. Observation of lesions using HE microscope The findings were three-fold: calcium oxalate crystal aggregation, renal tubular dilatation lesions, and chronic renal interstitial inflammatory cell infiltration.

[0126] The characteristics of the animal model were as follows:

[0127] After 4 weeks, the serum BUN content of the model animals increased significantly. There was no significant change in the P and CA contents in the blood, but the 24-hour urinary OX and CA excretion and the renal tissue CA content all increased significantly. The kidneys were macroscopically observed to be enlarged, and the cross sections were pale in color. When the kidney cross sections were touched with the hand, they had a clear rough texture of fine sand. The boundary between the renal cortex and the renal medulla was unclear. Compared with the normal group, the model group clearly showed aggregation of calcium oxalate crystals, swelling, degeneration, necrosis, and lumen expansion of tubular epithelial cells, and chronic renal interstitial inflammatory cell infiltration.

[0128] As shown in the figures of Figures 10 to 15, the following was observed under HE microscope. Compared with the normal group, the renal tubules in the model group were significantly dilated, numerous brownish yellow calcium oxalate crystals were observed, and inflammatory cells were infiltrated into the local renal interstitium. Compared with the model group, the renal tubule dilation was improved and the brownish yellow calcium oxalate crystals were significantly reduced in the medium and high dose groups. In addition, no obvious inflammatory cell infiltration in the renal interstitium was observed. In the low dose group, renal tubule dilation was improved and no obvious inflammatory cells were observed in the renal interstitium, but the brownish yellow calcium oxalate crystals were not significantly reduced. In the potassium sodium hydrogen citrate group, renal tubule dilation was improved and inflammatory cell infiltration in the renal interstitium was sometimes observed. In addition, the brownish yellow calcium oxalate crystals were not significantly reduced.

[0129] The evaluation sheet for renal lesions was as follows:

[0130] [Table 6] Note: No lesions are designated as -, moderate lesions as +, lesions as ++, and significant lesions as +++.

[0131] Based on the results of the above animal pharmacological tests, it has been fully proven that the medicinal ingredient extracts of Polygala gracilis (compounds obtained after the aqueous extract is eluted with 0-25% ethanol through a polyamide resin column) in the medium and high dose groups of the present invention from the three main indicators for the treatment of nephrolithiasis (aggregation of calcium oxalate crystals, renal tubular dilatation lesions, and chronic renal interstitial inflammatory cell infiltration) are all significantly superior to potassium sodium hydrogen citrate of the same quality. These drug efficacy levels have achieved the requirements of China Drug Registration Evaluation, indicating that it has excellent ability and market prospects in the treatment of urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug after surgical treatment of urolithiasis.

[0132] Benefits and Effects The drug of the present invention has significantly better effects than potassium sodium hydrogen citrate in typical test indicators such as calcium oxalate crystal aggregation, renal interstitial inflammatory cell infiltration, and renal tubular dilatation lesions, and these drug efficacy levels meet the requirements of China Drug Registration Evaluation, which shows its excellent ability and market prospects in treating urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and as an adjuvant drug after surgical treatment of urolithiasis.

[0133] Compared with potassium sodium hydrogen citrate, the most widely used clinical drug, the active ingredients (I) to (III) contained in the Polygala gracilis extract of the present invention do not contain sodium ions and potassium ions, and therefore do not lead to serious side effects (severe hyperkalemia, arrhythmia, and hypertension) as occurs with potassium sodium hydrogen citrate, making them safer.

[0134] Compared with other herbal medicines and drug extracts used in the treatment of diseases related to urinary tract stones, the pharmaceutical active ingredient extract of the present invention has simpler ingredients, clearer structures of the active ingredients, and more stable and controllable quality.

[0135] In addition, the pharmaceutical active ingredient extract of the present invention is preferably extracted from the stems and leaves of the aerial parts of Polygala gracilis, which is less expensive and more environmentally friendly, in order to avoid the problem of excessively long growth cycle of medicinal plants caused by extraction of the whole Polygala gracilis.

[0136] In other words, the drug of the present invention has obvious therapeutic effects on urolithiasis and other related diseases, few side effects (same as or less than those of potassium sodium hydrogen citrate, the existing main drug for urolithiasis), simple process, safe and effective, stable and controllable quality, meets the modern drug registration requirements, and has excellent medical and economic value.

Claims

1. Use of an aqueous or alcoholic extract of medicinal active ingredients extracted from the plant Polygala gracilis in the preparation of a medicament for the treatment or prevention of urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and for use as an adjuvant drug following surgical treatment of urolithiasis, comprising The medicinal active ingredient extract is (a) comprising one or more flavonol compounds represented by the following formulae (F-7K), (F-7Q), (F-74Q), (F-K), (F-Q), and (F-74K) as a first medicinal ingredient; 【Chemistry 1】 wherein R 1 is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Rha, -O-Glc-Glc-Api, -O-Gal-Glc-Api, -O-Glc-Gal-Api, and -O-Gal-Gal-Api; 【Chemistry 2】 wherein R 1 is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Glc, -O-Glc-Gal, -O-Glc-Api, -O-Glc-Rha, -O-Gal-Glc, -O-Gal-Gal, -O-Gal-Api, -O-Gal-Rha, -O-Glc-Glc-Api, -O-Gal-Glc-Api, -O-Gal-Gal-Api, -O-Glc-Gal-Api, -O-Gal-Gal-Api, -O-Gal-Rha-Gal, -O-Gal-Rha-Gal, -O-Gal-Rha-Glc, -O-Glc-Rha-Glc, and -O-Glc-Rha-Gal; 【Chemistry 3】 wherein R 1 is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Api, and -O-Gal-Api; 【Chemistry 4】 wherein R is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Rha, and -O-Gal-Rha; 【Chemistry 5】 wherein R is selected from the group consisting of -OH, -O-Glc, -O-Gal, -O-Glc-Rha, and -O-Gal-Rha; 【Chemistry 6】 wherein R is selected from the group consisting of -OH, -O-Gal, and -O-Gal-Api; the total content of the flavonol compound which is the first medicinal ingredient (a) is 75 to 100% of the total extract of Polygala gracilis; Here, the content (%) of a component is the HPLC% content calculated using the HPLC integral area normalization method according to a method commonly used in the art. Use of an aqueous or alcoholic extract of medicinal active ingredients extracted from the plant Polygala gracilis.

2. Use of a water extract or alcohol extract of a medicinal medicinal ingredient extracted from the plant Polygala gracilis described in claim 1, further comprising (b) a chitosan compound selected from one or more of the following formulae (II-1), (II-2), and (II-3) as a second medicinal ingredient. 【Chemistry 7】 【Chemistry 8】 【Chemistry 9】

3. Use of a water extract or alcohol extract of a medicinal medicinal ingredient extracted from the plant Polygala gracilis described in claim 1 or 2, further comprising (c) a glycolipid compound selected from one or more of the following formulae (III-1) or (III-2) as a third medicinal ingredient. 【Chemistry 10】 【Chemistry 11】

4. 3. Use of an aqueous or alcoholic extract of a medicinal active ingredient extracted from the plant Polygala gracilis according to claim 1 or 2, wherein the first active ingredient (a) is one selected from the following compounds: 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】

5. 2. Use of any one of the following compounds in the preparation of a medicament for the treatment or prevention of urolithiasis, urinary tract infection, or kidney damage caused by urolithiasis, and for use as an adjuvant drug following surgical treatment of urolithiasis: 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】

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