A composition for tonifying kidney and its preparation method

Abstract

The application provides a kidney-tonifying composition and a preparation method thereof, and belongs to the technical field of traditional Chinese medicine extracts. The preparation method comprises the following steps: S1, crushing Tribulus terrestris and screening to obtain Tribulus terrestris powder; S2, taking fresh Cistanche deserticola and sodium chloride, crushing and grinding together, heating under nitrogen protection, adding methanol, concentrated hydrochloric acid and the Tribulus terrestris powder obtained in step S1, refluxing, filtering to obtain a crude extract; S3, concentrating the crude extract obtained in step S2, adjusting the pH to obtain a concentrated solution; S4, column chromatography separation of the concentrated solution obtained in step S3, concentrating and drying the eluent to obtain a kidney-tonifying traditional Chinese medicine extract. The application further provides a kidney-tonifying composition, which is prepared from the kidney-tonifying traditional Chinese medicine extract and Myristicae Semen, Polygalae Radix, Morus alba fruit, and Radix Glycyrrhizae. The composition has good application prospect due to the good kidney-tonifying effect of the monarch, the ministers and the aids.

Description

Technical Field

[0001] The present invention belongs to the technical field of traditional Chinese medicine compositions, and particularly relates to a kidney-tonifying and strengthening composition and a preparation method thereof. Background Art

[0002] In recent years, with increasing work pressure, the number of patients suffering from kidney deficiency has also been increasing year by year. Clinically, symptoms of kidney yang deficiency include pale or dark complexion, weakness in the waist and knees, lack of energy, cold hands and feet, fear of cold and wind, loose stools and diarrhea, insomnia and irritability, impotence and premature ejaculation in men, low libido, frequent urination and nocturnal emissions, uterine cold and infertility in women, and menstrual irregularities. Existing kidney-tonifying and yang-strengthening products are primarily divided into Traditional Chinese Medicine (TCM) and Western medicine. Among Western medicines, chemically synthesized drugs, such as sildenafil citrate, can address male sexual dysfunction to a certain extent, but they struggle to address the root cause of the problem through holistic health care. Traditional Chinese Medicine (TCM), with its reliable efficacy, minimal toxicity and side effects, and safe clinical application, offers unique advantages for improving and preventing these symptoms.

[0003] Tribulus terrestris Fructus Tribuli It is mainly produced in Henan, Hebei, Shandong, Anhui, Jiangsu, Sichuan, Shaanxi and other places in my country. The medicinal base of Chinese medicine is the fruit part. The multiple fruit of Tribulus terrestris is mostly composed of 5 lobes, which are radially arranged in a prismatic spherical shape. The commercial fruit peel is hard and wooden, containing 3-4 seeds. The seeds are oval, slightly flat and oily. The smell is faint and the taste is bitter. The fruit of Tribulus terrestris contains tribulus glycoside, kaempferol, kaempferol-3-glucoside, kaempferol-3-rutinoside, quercetin and other ingredients. The seed oil contains tribulamide, N-trans-p-hydroxyphenylethyl caffeamide, physcion methyl ether and other ingredients. Modern pharmacological studies have shown that Tribulus terrestris has anti-hypertensive, anti-myocardial ischemia, anti-aging and sexual enhancement effects.

[0004] Cistanche deserticola Herba Cistanches Cistanche deserticola is primarily produced in Inner Mongolia, Ningxia, Gansu, and Xinjiang. Its medicinal base is its fleshy stem. Its stem is oblate, slightly curved, and has a brown or grayish-brown surface. It is densely covered with fleshy, diamond-shaped or triangular scales arranged in a shingle-like pattern. It has a faint aroma and a sweet, slightly bitter taste. Its stem contains cistanchesides A, B, C, and H, eugenol, 2'-acetyleugenol, echinoside, (2,5-dioxo-4-imidazolidinyl)amino acid, betaine, daucin, 8-epilogous acid, β-sitosterol, and N,N-dimethylglycine methyl ester. It also contains a variety of amino acids, polysaccharides, and other nutrients. Modern pharmacological research has shown that Cistanche deserticola has immune-enhancing, endocrine-regulating, metabolic-stimulating, anti-aging, and laxative effects.

[0005] Chinese patent CN103405617B discloses a Chinese herbal composition for tonifying the kidney and strengthening yang, and its preparation method. The raw materials are composed of epimedium, bullwhip, psoralea corylifolia, Morinda officinalis, red ginseng, astragalus, cinnamon bark, leek seeds, walnut kernel, Rehmannia root, wolfberry fruit, Rehmannia root, silkworm cocoon, Chinese angelica, jujube, and Sichuan aconite. These raw materials are purified by a specific process to obtain the Chinese herbal composition. However, the raw material formula of this composition is large, and the inclusion of the animal-derived drugs bullwhip and silkworm cocoon increases the cost.

[0006] Chinese Patent No. 105497496B discloses a traditional Chinese medicine composition for improving male sexual function, its preparation method, and its use. The raw materials are composed of ginseng, schisandra chinensis, ophiopogon japonicus, wolfberry fruit, and salvia miltiorrhiza. The formula is simple and relatively cost-effective, but animal experiments have shown limited effectiveness in improving sexual function.

[0007] On the other hand, Chinese medicine extraction technology helps to improve the effects of Chinese medicine from the perspective of enriching the effective ingredients of Chinese medicine, and at the same time can remove unnecessary unknown components in Chinese medicinal materials. It is the only way to achieve better application effects of Chinese medicine and Chinese medicine compositions.

[0008] Chinese Patent No. 102441040A discloses a method for preparing total phenylethanoid glycosides from Cistanche deserticola, comprising the steps of pulverizing the Cistanche deserticola, extracting it, adsorbing it with an adsorption resin, eluting it, and concentrating it. This method can prevent the decomposition and oxidation of phenylethanoid glycosides from Cistanche deserticola. However, this method does not disclose the kidney-strengthening and male sexual function-improving effects of the extract obtained from this method.

[0009] Chinese Patent 116966213A discloses a method for extracting tribulus saponins from Tribulus terrestris and its application, addressing the issues of existing single extraction methods and the low purity of tribulus saponins. The method includes drying the tribulus terrestris, pulverizing it, extracting it with alcohol, extracting it with a mixed solvent, and recrystallizing it. However, the method does not disclose the extract's ability to strengthen the kidneys and improve male sexual function.

[0010] In summary, it is difficult in the prior art to provide a preparation method for simultaneously extracting effective ingredients from the Chinese medicinal raw materials of Tribulus terrestris and Cistanche deserticola, and making the obtained Chinese medicinal extract have better kidney-tonifying and kidney-strengthening effects. Summary of the Invention

[0011] In view of this, the existing technology is difficult to provide a preparation method for extracting effective ingredients from the Chinese medicinal raw materials of Tribulus terrestris and Cistanche deserticola at the same time, and making the obtained Chinese medicinal extract have better kidney-tonifying and kidney-strengthening effects. The purpose of the present invention is to provide a preparation method, product and application of a kidney-strengthening Chinese medicinal extract.

[0012] To achieve the purpose of the present invention, on the one hand, the present invention provides a method for preparing a kidney-strengthening Chinese medicine extract, comprising the following steps:

[0013] S1, crushing and sieving Tribulus terrestris L. to obtain Tribulus terrestris L. powder;

[0014] S2, taking fresh Cistanche and granular sodium chloride, crushing and grinding together, heating under nitrogen protection, then adding methanol, concentrated hydrochloric acid, and the Tribulus terrestris L. powder obtained in step S1, refluxing, and filtering to obtain a crude extract;

[0015] S3, concentrating the crude extract obtained in step S2 and adjusting the pH to obtain a concentrated solution;

[0016] S4, column chromatography separation of the concentrated solution obtained in step S3, concentration and drying of the eluent to obtain a kidney-strengthening traditional Chinese medicine extract.

[0017] Preferably, in step S1, the amount of Tribulus terrestris L. is 23-32 parts by mass.

[0018] More preferably, in step S1, the amount of Tribulus terrestris L. is 28 parts by mass.

[0019] Preferably, in step S2, the amount of fresh Cistanche is 100-120 parts by mass.

[0020] More preferably, in step S2, the amount of fresh Cistanche is 110 parts by mass.

[0021] Preferably, in step S2, the amount of sodium chloride is 45-55 parts by mass.

[0022] More preferably, in step S2, the amount of sodium chloride is 50 parts by mass.

[0023] Preferably, in step S2, the amount of methanol is 150-250 parts by mass.

[0024] More preferably, in step S2, the amount of methanol is 200 parts by mass.

[0025] In step S2, the concentrated hydrochloric acid is commercially available concentrated hydrochloric acid with a hydrogen chloride mass fraction of 36%-38%.

[0026] Preferably, in step S2, the amount of concentrated hydrochloric acid is 0.8-1.5 parts by mass.

[0027] More preferably, in step S2, the amount of concentrated hydrochloric acid is 1 part by mass.

[0028] Preferably, in step S2, the sodium chloride is granular sodium chloride.

[0029] More preferably, the particle size of the granular sodium chloride is greater than or equal to 1 mm.

[0030] More preferably, and as an example of the present application, the particle size of the granular sodium chloride is 2-2.8 mm.

[0031] Preferably, in step S2, the grinding time is 10-45 min.

[0032] More preferably, in step S2, the grinding time is 30 min.

[0033] Preferably, in step S2, the heating temperature under nitrogen protection is 65-85℃.

[0034] More preferably, in step S2, the heating temperature under nitrogen protection is 75℃.

[0035] Preferably, in step S2, the heating time under nitrogen protection is 20-50 min.

[0036] More preferably, in step S2, the heating time under nitrogen protection is 30 min.

[0037] In another aspect, the present application provides a kidney-strengthening traditional Chinese medicine extract prepared by the above preparation method.

[0038] In still another aspect, the present application provides a kidney-tonifying composition prepared from the above kidney-strengthening traditional Chinese medicine extract, Myristica fragrans, Polygala tenuifolia, Morus alba fruit, and Radix glycyrrhizae; wherein the kidney-strengthening traditional Chinese medicine extract is prepared by the above preparation method.

[0039] Preferably, the composition is prepared from 14-23 parts by mass of the kidney-strengthening traditional Chinese medicine extract, 10-20 parts by mass of Myristica fragrans, 5-9 parts by mass of Polygala tenuifolia, and 3-8 parts by mass of Radix glycyrrhizae; wherein the raw materials of the kidney-strengthening traditional Chinese medicine extract are 23-32 parts by mass of Tribulus terrestris and 100-120 parts by mass of fresh Herba cistanche.

[0040] More preferably, the composition is prepared from 19 parts by mass of Myristica fragrans, 15 parts by mass of Polygala tenuifolia, 7 parts by mass of Morus alba fruit, and 5 parts by mass of Radix glycyrrhizae; wherein the raw materials of the kidney-strengthening traditional Chinese medicine extract are 28 parts by mass of Tribulus terrestris and 110 parts by mass of fresh Herba cistanche.

[0041] In still another aspect, the present application provides a preparation method of the above composition, comprising the following steps:

[0042] T1, mixing Myristica fragrans with an organic solvent, heating to reflux, filtering to obtain a filtrate, concentrating, and drying to obtain a Myristica fragrans extract;

[0043] T2, drying, decocting, filtering, and combining the filtrates of Polygala tenuifolia, Morus alba fruit, and Radix glycyrrhizae to obtain a total filtrate;

[0044] T3, mixing the kidney-strengthening Chinese medicine extract, the myristica extract obtained in step T1, and the total filtrate obtained in step T2, concentrating, to obtain a composition.

[0045] Preferably, in step T1, the organic solvent is selected from at least one of n-hexane, cyclohexane, acetone, and ethanol.

[0046] More preferably, in step T1, the organic solvent is a mixture of ethanol and cyclohexane.

[0047] Still more preferably, and as an example of the present application, the mixture of ethanol and cyclohexane is specifically a mixture of ethanol and cyclohexane in a volume ratio of 2.5:1.

[0048] Preferably, in step T1, the mass ratio of the organic solvent to the myristica is 100:17-23.

[0049] Preferably, in step T1, the concentrating is specifically rotary evaporation.

[0050] Preferably, in step T1, the drying is specifically removal of the organic solvent by drying.

[0051] Preferably, in step T2, the decocting is specifically adding 3-8 times the mass of water to the prepared mixture, and decocting for 3-5 hours.

[0052] In still another aspect, the present application provides a medicine, the raw material of which is the kidney-strengthening Chinese medicine extract prepared by the above method or / and the above composition or / and the composition prepared by the above method.

[0053] The medicine can be prepared into dosage forms for administration, such as pills, capsules, granules, oral liquids, powders, tablets, lozenges, and sugar lozenges, and suitable excipients in the art can be selected for different dosage forms.

[0054] The excipients used can be solid, liquid, or gas. Examples of solid excipients include lactose, white clay, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid excipients include sugar syrup, peanut oil, olive oil, and water. Examples of gaseous excipients include carbon dioxide and nitrogen.

[0055] In preparing the composition of oral dosage forms, any convenient pharmaceutical medium can be used. For example, water, ethanol, oil, alcohol, flavoring agents, preservatives, colorants, etc. can be used to form oral liquid preparations, such as suspensions, emulsions, and solutions; while carriers, such as starch, sugar, microcrystalline cellulose, diluents, granulating agents, emulsifying agents, lubricants, binders, disintegrating agents can be used to form oral solid preparations, such as powders, capsules, and tablets. Due to their ease of administration, tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers. Standard aqueous or non-aqueous techniques can be selected for coating of tablets.

[0056] Tablets containing the compositions of the present invention can be prepared by tableting or molding, optionally with the use of one or more auxiliary ingredients or adjuvants. Compressed tablets can be prepared by compressing the active ingredient in a free-flowing form (such as a powder or granules) in a suitable machine, optionally mixed with a binder, lubricant, inert diluent, surfactant, or dispersant. Molded tablets can be molded in a suitable machine, i.e., a mixture of powdered compounds moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of active ingredient. For example, a formulation intended for oral administration to humans may contain about 0.5 mg to about 5 g of active drug, mixed with an appropriate and convenient carrier material, which may account for about 5% to 95% of the total composition. Unit dosage forms typically contain about 1 mg to about 2 g of active ingredient, typically 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg.

[0057] Pharmaceutical compositions suitable for parenteral administration of the present invention can be prepared as aqueous solutions or suspensions of the active compound. Suitable surfactants, such as hydroxypropylcellulose, can be included. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, and oil mixtures thereof. In addition, preservatives can be added to prevent the harmful growth of microorganisms.

[0058] Medicine of the present invention can be the form that is suitable for topical use, for example aerosol, cream, ointment, lotion, powder or the like.In addition, composition can be suitable form and be used for transdermal administration device.Can use Chinese medicine composition of the present invention, prepare these prescriptions by conventional processing method.For example, by mixing hydrophilic material and water, and about 5wt% to about 10wt% compound, prepare cream or ointment with required consistency.

[0059] The medicament of the present invention can be in a form suitable for rectal administration, wherein the carrier is a solid. It is best to prepare the mixture into unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. Suppositories can be prepared by first forming a composition containing a softened or melted carrier, followed by cooling and shaping in a mold.

[0060] In addition to the above-mentioned carrier components, the above-mentioned pharmaceutical preparations may include (if applicable) one or more additional carrier components, such as diluents, buffers, flavoring agents, binders, surfactants, thickeners, lubricants, preservatives (including antioxidants), etc. In addition, other excipients, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc., colorants and flavoring agents, etc., may be added. The preparation is made isotonic with the blood of the intended recipient. The components containing the Chinese medicine composition of the present invention can also be prepared in the form of a powder or concentrate.

[0061] Preferably, the dosage form of the medicine comprises ointment, pill, granule, oral liquid, tablet, lozenge, capsule, powder, syrup, tincture.

[0062] In still another aspect, the present application provides the above-mentioned kidney-tonifying traditional Chinese medicine extract and its preparation method, the above-mentioned composition and its preparation method, and the above-mentioned kidney-tonifying medicine in the production of kidney-tonifying products.

[0063] Preferably, the kidney-tonifying product is a kidney-tonifying medicine.

[0064] The performance of the traditional Chinese medicine components used in the present application is further explained as follows.

[0065] Triadax Spinosus:

[0066]

Synonyms

[0067]

Origin

[0068]

Original Plant

[0069] 2. Triadax Spinosus T. cistoides L.

[0070]

Harvesting and Processing

[0071]

Medicine Property

[0072]

Function and Main Treatment

[0073] Cistanche:

[0074]

Synonyms

[0075]

Origin

[0076]

Original Plant

[0077]

Harvesting and Processing

[0078]

Property

[0079]

Function and Indication

[0080] Myristica fragrans Houtt.

[0081]

Synonym

[0082]

Origin

[0083]

Original Plant

[0084]

Harvesting and Processing

[0085]

Property

[0086]

Function and Indication

[0087] Polygala sibirica L.

[0088] ​

Syn.

[0089]

Source

[0090]

Original Plant

[0091] 2. Polygala sibirica L. P. sibirica

[0092]

Harvesting Processing

[0093]

Property

[0094] Compendium of Materia Medica records: “Polygala enters the kidney channel of the lesser yin foot, and is not a heart channel drug. Its function is to strengthen the will and ease the mind, and treat forgetfulness. Since essence and will are both stored in the kidney channel.”

[0095]

Function-Indications

[0096] Mulberry Fruit:

[0097]

Syn.

[0098]

Source

[0099]

Original Plant

[0100]

Harvesting Processing

[0101]

Property

[0102]

Function-Indications

[0103] Licorice:

[0104] [Synonyms] Meicao, Migan (Ben Cao), Micao, Heigencao (Bielu), Guolao (Bencao Jing Jizhu), Lingtong (Jisizhu), Fencao (Qunfangpu), Tiancao (Chinese Medicinal Plants), Tiangenzi (Zhongyaozhi), Bangcao (Heilongjiang Traditional Chinese Medicine).

[0105] [Origin] The roots and rhizomes of Glycyrrhiza glabra, Glycyrrhiza glabra and Glycyrrhiza inflata, all plants of the genus Glycyrrhiza in the Leguminosae family.

[0106]

Original Plant

[0107] 2. Licorice G. glabra L.

[0108] 3. Licorice root G. inflata Batal.

[0109] [Harvesting and processing] Dig up in August-September, remove the rhizome, stem base and fibrous roots, cut into sections of appropriate length, sun-dry until half-dried, tie into small bundles, and sun-dry again until completely dry.

[0110] [Processing] The licorice used in the present invention is processed licorice. Roasted licorice is also known as honey licorice. The processing method is as follows:

[0111] Dilute refined honey with an appropriate amount of boiling water, add licorice slices, mix well, let it simmer for a while, place in a hot pot, and stir-fry over low heat until the surface turns dark yellow and is no longer sticky. Remove and let cool. For every 100 kg of licorice slices, use 25 kg of refined honey. Honey licorice resembles licorice root, has a dark yellow, slightly glossy surface, is slightly sticky, and has a sweet taste. It is used to tonify the middle qi and relieve acute pain. According to the "Medicinal Properties Micro-Yun", "(Licorice) is sweet and warm, and can eliminate severe heat. Therefore, when used raw, it calms qi, replenishes spleen and stomach deficiency, and significantly relieves heart fire. When roasted, it warms qi, replenishes the three burners, dispels superficial cold, eliminates pathogenic heat, relieves sore throat, soothes vital energy, and nourishes yin and blood."

[0112]

Properties

[0113] [Functions and Indications] It can soothe the stomach and relieve urgency, moisten the lungs and detoxify, and harmonize various medicines. Roasted, it treats spleen and stomach deficiency, fatigue, poor appetite, abdominal pain and loose stools, cramps and pain in the limbs, palpitations and restlessness, and pulmonary tuberculosis and cough. Raw, it treats sore throat, carbuncle swelling, fetal toxins in children, and drug and food poisoning.

[0114] Compared with the prior art, the present invention has the following beneficial effects:

[0115] (1) The preparation method of the kidney-strengthening traditional Chinese medicine extract provided by the application can efficiently extract effective components with kidney-strengthening functions in two kinds of traditional Chinese medicinal materials, i.e., Tribulus terrestris and Cistanche, and compared with conventional extraction methods, the traditional Chinese medicine extract extracted by the method has good kidney-strengthening effects.

[0116] (2) In the raw traditional Chinese medicinal materials of the kidney-tonifying and kidney-strengthening composition provided by the application, Cistanche fills up kidney yang and increases essence and blood, and is the monarch drug; Polygala improves kidney and yang and easily essence, Myristica removes cold and dampness, warms and supplements the spleen and stomach to warm the kidney, Tribulus terrestris calms the liver and benefits essence, cools and nourishes blood, benefits and supplements yin, and is the minister drug; mulberry fruit nourishes yin and blood, benefits the kidney and essence, clears virtual fire and stops virtual wind, and is the assistant drug; and the fried licorice root harmonizes the middle and relieves urgency, and is the ministerial drug. The whole formula assists in supplementing kidney yang, and has the effects of supplementing blood and nourishing yin, dispersing cold and resolving deficiency.

[0117] (3) The composition is proved by animal experiments to have significant effects of tonifying the kidney and strengthening the kidney and improving sexual function, and has wide application prospects. DETAILED DESCRIPTION

[0118] The following non-limiting examples can enable those skilled in the art to more fully understand the present application, but do not limit the present application in any way. The following content is only an exemplary description of the scope of the present application, and those skilled in the art can make various changes and modifications to the application disclosed herein, which should also belong to the scope of the present application.

[0119] The present application is further described below in the form of specific examples. The various chemical reagents used in the examples of the present application are obtained through conventional commercial channels unless otherwise specified. If not otherwise specified, the contents described below are mass contents, and the parts of the raw materials are mass parts. If not otherwise specified, it is understood to be carried out at room temperature.

[0120] Example 1

[0121] 1. Preparation of traditional Chinese medicine extract

[0122] S1, dry Tribulus terrestris 28 parts are crushed using a high-throughput crusher, and sieved through a pharmacopoeia No. 3 sieve to obtain Tribulus terrestris powder.

[0123] S2, take fresh Cistanche 110 parts, granular sodium chloride (average particle size 2-2.8 mm) 50 parts, and pound together into a paste, then grind for 30 min, and transfer into a flask. Under nitrogen protection, heat to 75℃ for 30 min, then add methanol 200 parts, concentrated hydrochloric acid 1 part, and the Tribulus terrestris powder obtained in step S1, heat and maintain reflux for 3 hours. After cooling to room temperature, filter to obtain the filtrate as a crude extract.

[0124] S3, the crude extract prepared in step S2 is concentrated to a volume of 0.2 times the original volume by room temperature distillation under reduced pressure, and the pH is adjusted to 6-8 using sodium bicarbonate to obtain a concentrated solution.

[0125] S4, the concentrated solution prepared in step S3 is separated by column chromatography, and eluted with an ethanol-water solution. The eluate is concentrated by a rotary evaporator at 30°C and dried at 15°C to obtain a traditional Chinese medicine extract. The chromatography column used is a D101 macroporous resin chromatography column.

[0126] 2. Preparation of the composition.

[0127] T1, 19 parts by mass of nutmeg is mixed with 100 parts by mass of an organic solvent, heated to reflux for 8 hours, filtered to obtain the filtrate, rotary evaporated at 50°C, and dried to obtain a nutmeg extract. The organic solvent is a mixture of ethanol and cyclohexane in a volume ratio of 2.5:1.

[0128] T2, 15 parts of radix polygalae, 7 parts of fructus mori, and 5 parts of honeyed licorice are vacuum dried at 45°C for 12 hours, ground by a pulverizer to pass through a No. 3 sieve, and mixed with 8 times the mass of water to decoct for 5 hours. The filtrate is obtained by suction filtration. The residue is mixed with 5 times the mass of water to decoct for 4 hours, and the filtrate is obtained by suction filtration. Further, the residue is mixed with 3 times the mass of water to decoct for 3 hours, and the filtrate is obtained by suction filtration. The filtrates obtained by the three decoctions are combined to obtain a total filtrate.

[0129] T3, the traditional Chinese medicine extract obtained in step S4 of the preparation of the traditional Chinese medicine extract (110 parts of fresh nutmeg and 28 parts of dried tribulus terrestris) is mixed with the nutmeg extract obtained in step T1 and the total filtrate obtained in step T2, and concentrated to a relative density of 1.4 to obtain a clear paste, which is the composition.

[0130] Example 2

[0131] 1. Preparation of the traditional Chinese medicine extract.

[0132] S1, 23 parts of dried tribulus terrestris is ground by a high-throughput pulverizer to pass through a No. 3 sieve to obtain tribulus terrestris powder.

[0133] S2, 120 parts of fresh cistanche and 55 parts of coarse salt (average particle size greater than or equal to 1 mm) are mashed together to a paste, ground for 30 min, and transferred to a flask. Under nitrogen protection, heated to 75°C for 30 min, then added 200 parts of methanol, 1 part of concentrated hydrochloric acid, and the tribulus terrestris powder obtained in step S1, and heated to reflux for 3 hours. After cooling to room temperature, the filtrate is obtained by suction filtration as a crude extract.

[0134] S3, the crude extract prepared in step S2 is concentrated to a volume of 0.2 times the original volume by room temperature distillation under reduced pressure, and the pH is adjusted to 6-8 using sodium bicarbonate to obtain a concentrated solution.

[0135] S4, column chromatography separation of the concentrated solution prepared in step S3, eluted with ethanol-water solution, the eluate was collected, the eluate was concentrated by rotary evaporator 30℃, 15℃ drying, to obtain the extract of traditional Chinese medicine.

[0136] 2. Preparation of the composition.

[0137] T1, 23 parts by mass of nutmeg was mixed with 100 parts by mass of organic solvent, heated to reflux for 8 hours, filtered to take the filtrate, 50℃ rotary evaporation, drying, to obtain the nutmeg extract. The organic solvent is a mixture of ethanol, cyclohexane in a volume ratio of 2.5:1.

[0138] T2, 10 parts of polygala, 5 parts of mulberry fruit, 8 parts of fried licorice were vacuum dried at 45℃ for 12 hours, crushed by a pulverizer, and passed through a No. 3 sieve. The residue was mixed with 8 times the mass of water and decocted for 5 hours. The filtrate was obtained by filtration. The residue was mixed with 5 times the mass of water and decocted for 4 hours. The filtrate was obtained by filtration. Further, the residue was mixed with 3 times the mass of water and decocted for 3 hours. The filtrate was obtained by filtration. The filtrates obtained by the three decoctions were combined to obtain the total filtrate.

[0139] T3, the traditional Chinese medicine extract obtained in step S4 of the preparation of the traditional Chinese medicine extract (120 parts of fresh nutmeg, 23 parts of dried tribulus terrestris) was mixed with the nutmeg extract obtained in step T1 and the total filtrate obtained in step T2, concentrated to a relative density of 1.4 to obtain a clear paste, which is the composition.

[0140] Compared with Example 1, the amounts of tribulus terrestris, cistanche, nutmeg, polygala, mulberry fruit and fried licorice were adjusted.

[0141] Example 3

[0142] 1. Preparation of the traditional Chinese medicine extract.

[0143] S1, dry tribulus terrestris 32 parts was crushed by a high-throughput pulverizer, and passed through a No. 3 sieve to obtain tribulus terrestris powder.

[0144] S2, fresh cistanche 100 parts, coarse salt (average particle size greater than or equal to 1 mm) 45 parts were mashed together into a paste, ground for 30 min, and transferred into a flask. Under nitrogen protection, heated to 75℃ for 30 min, then added methanol 200 parts, concentrated hydrochloric acid 1 part, and tribulus terrestris powder obtained in step S1. The mixture was heated and refluxed for 3 hours. After cooling to room temperature, the filtrate was filtered to obtain the crude extract.

[0145] S3, the crude extract prepared in step S2 was subjected to room temperature vacuum distillation, concentrated to a volume of 0.1-0.35 times that of the original, and the pH was adjusted to 6.5-8 using sodium bicarbonate to obtain a concentrated solution.

[0146] S4, column chromatography separation of the concentrated solution prepared in step S3, eluted with ethanol-water solution, the eluate was collected, the eluate was concentrated by rotary evaporator 30℃, 15℃ drying, get traditional Chinese medicine extract.

[0147] 2. Preparation of the composition.

[0148] T1, 17 parts by mass of nutmeg and 100 parts by mass of organic solvent were mixed and heated to reflux for 8 hours. The filtrate was filtered and concentrated by rotary evaporation at 50℃. The dried nutmeg extract was obtained. The organic solvent was a mixture of ethanol and cyclohexane in a volume ratio of 2.5:1.

[0149] T2, 20 parts of polygala, 5 parts of mulberry fruit, 8 parts of fried licorice were vacuum dried at 45℃ for 12 hours, and then crushed through a 400 mesh sieve. The mixture was mixed with 8 times the mass of water and boiled for 5 hours. The filtrate was obtained by filtration. The residue was mixed with 5 times the mass of water and boiled for 4 hours. The filtrate was obtained by filtration. Further, the residue was mixed with 3 times the mass of water and boiled for 3 hours. The filtrate was obtained by filtration. The filtrates obtained by the three times of boiling were combined to obtain the total filtrate.

[0150] T3, the traditional Chinese medicine extract obtained in step S4 of the preparation of the traditional Chinese medicine extract (100 parts of fresh nutmeg, 32 parts of dried tribulus terrestris) was mixed with the nutmeg extract obtained in step T1 and the total filtrate obtained in step T2. Concentrate to relative density 1.4 to obtain clear paste, which is the composition.

[0151] Compared with example 1 and example 2, the amount of tribulus terrestris, cistanche, nutmeg, polygala, mulberry fruit and fried licorice was adjusted.

[0152] Example 4

[0153] 1. Preparation of traditional Chinese medicine extract.

[0154] S1, dry tribulus terrestris 28 parts was crushed by high throughput crusher, and then passed through pharmacopoeia No. 3 sieve to obtain tribulus terrestris powder.

[0155] S2, take fresh cistanche 110 parts, granular sodium chloride (average particle size 2-2.8mm) 50 parts, together to mash to paste, then grind for 10 min, transfer to flask. Under nitrogen protection, heat to 85℃ for 20 min, then add methanol 250 parts, concentrated hydrochloric acid 1.5 parts, tribulus terrestris powder obtained in step S1, heat to reflux for 3 hours. After cooling to room temperature, the filtrate was filtered and obtained as the crude extract.

[0156] S3, the crude extract prepared in step S2 was subjected to room temperature vacuum distillation, concentrated to 0.2 times the original volume, and the pH was adjusted to 6-8 with sodium bicarbonate to obtain the concentrated solution.

[0157] S4, the concentrated solution prepared in step S3 is subjected to column chromatography separation, elution is performed using an ethanol-water solution, the eluate is collected, the eluate is concentrated by a rotary evaporator at 30°C, and dried at 15°C to obtain a traditional Chinese medicine extract. The chromatography column used is a D101 macroporous resin chromatography column.

[0158] 2. Preparation of the composition.

[0159] T1, 19 parts by mass of mace are mixed with 100 parts by mass of n-hexane, heated to reflux for 8 hours, filtered to take the filtrate, rotary evaporated at 50°C, and dried to obtain a mace extract.

[0160] T2, 15 parts of polygala, 7 parts of mulberry fruit, and 5 parts of fried licorice are vacuum dried at 45°C for 12 hours, pulverized by a pulverizer to pass through a 400-mesh sieve, mixed with 8 times the mass of water to decoct for 5 hours, and filtered to take the filtrate. The filter residue is mixed with 5 times the mass of water to decoct for 4 hours, and filtered to take the filtrate. Further, the filter residue is mixed with 3 times the mass of water to decoct for 3 hours, and filtered to take the filtrate. The filtrates obtained in the three decocting steps are combined to obtain a total filtrate.

[0161] T3, the traditional Chinese medicine extract obtained in step S4 of the preparation of the traditional Chinese medicine extract (110 parts of fresh mace and 28 parts of dried tribulus terrestris) is mixed with the mace extract obtained in step T1 and the total filtrate obtained in step T2, concentrated to a relative density of 1.4 to obtain a clear extract, which is the composition.

[0162] Example 5

[0163] 1. Preparation of the traditional Chinese medicine extract.

[0164] S1, 28 parts of dried tribulus terrestris is pulverized by a high-throughput pulverizer to pass through a No. 3 pharmacopoeia sieve to obtain tribulus terrestris powder.

[0165] S2, 110 parts of fresh cistanche and 50 parts of granular sodium chloride (average particle size 2-2.8 mm) are mashed together to a paste, and then ground for 45 min and transferred into a flask. Under nitrogen protection, heated to 65°C for 50 min, then 150 parts of methanol, 0.8 parts of concentrated hydrochloric acid, and the tribulus terrestris powder obtained in step S1 are added, heated to reflux for 3 hours. After cooling to room temperature, the filtrate is filtered to obtain a crude extract.

[0166] S3, the crude extract prepared in step S2 is subjected to room temperature reduced pressure distillation, concentrated to a volume of 0.2 times that of the original, and the pH is adjusted to 6-8 using sodium bicarbonate to obtain a concentrated solution.

[0167] S4, the concentrated solution prepared in step S3 is subjected to column chromatography separation, elution is performed using an ethanol-water solution, the eluate is collected, the eluate is concentrated by a rotary evaporator at 30°C, and dried at 15°C to obtain a traditional Chinese medicine extract. The chromatography column used is a D101 macroporous resin chromatography column.

[0168] 2. Preparation of the composition.

[0169] T1, 19 parts of Myristica fragrans by mass was mixed with 100 parts of acetone, heated to reflux for 8 hours, filtered to take the filtrate, rotary evaporated at 50°C, dried to obtain Myristica fragrans extract.

[0170] T2, 15 parts of Polygala tenuifolia, 7 parts of Morus alba, and 5 parts of Radix Glycyrrhizae were vacuum dried at 45°C for 12 hours, ground by a pulverizer to pass through a 400-mesh sieve, mixed with 8 times the mass of water to decoct for 5 hours, and filtered to take the filtrate. The residue was mixed with 5 times the mass of water to decoct for 4 hours, and filtered to take the filtrate. Further, the residue was mixed with 3 times the mass of water to decoct for 3 hours, and filtered to take the filtrate. The filtrates obtained in the three decocting processes were combined to obtain the total filtrate.

[0171] T3, the Chinese medicinal extract obtained in step S4 (110 parts of fresh Myristica fragrans and 28 parts of dried Tribulus terrestris) was mixed with the Myristica fragrans extract obtained in step T1 and the total filtrate obtained in step T2, concentrated to a relative density of 1.4 to obtain a clear extract, which was the composition.

[0172] Comparative Example 1

[0173] Compared with Example 1, in step S1, the granular sodium chloride (average particle size 2-2.8 mm) was replaced with the same mass of quartz sand (average particle size 2.5 mm), and the rest was the same.

[0174] Comparative Example 2

[0175] Compared with Example 1, in step S1, the granular sodium chloride (average particle size 2-2.8 mm) was replaced with powdered sodium chloride (particle size less than 300 μm), and the rest was the same.

[0176] Comparative Example 3

[0177] Compared with Example 1, in step S1, the granular sodium chloride (average particle size 2-2.8 mm) was replaced with 10 parts, and the rest was the same.

[0178] Comparative Example 4

[0179] Compared with Example 1, in step S2, the nitrogen protection was replaced with air, and the rest was the same.

[0180] Comparative Example 5

[0181] Compared with Example 1, in step S2, the addition of 200 parts of methanol and 2 parts of concentrated hydrochloric acid was replaced with the addition of 200 parts of methanol and 2 parts of glacial acetic acid, and the rest was the same.

[0182] Comparative Example 6

[0183] Compared with Example 1, the polygala in step T2 is replaced by the same mass fraction of ophiopogon, and the rest is the same.

[0184] Comparative Example 7

[0185] 1. Preparation of Chinese medicine extract

[0186] 28 parts of dried Tribulus terrestris L. were crushed, passed through a No. 3 pharmacopoeia sieve, mixed with 110 parts of fresh Cistanche deserticola Y.C.Ma and 202 parts of water, and heated to decoct. After being cooled to room temperature, the filtrate was obtained by filtration. The residue was decocted twice again, and the amount of water used for each decoction was 5 times the mass of the solid material. The filtrates were combined, concentrated, and separated by column chromatography. Ethanol-water solution was used for elution, and the eluate was collected. The eluate was concentrated by a rotary evaporator at 30°C and dried at 15°C to obtain the Chinese medicine extract. The chromatography column used was a D101 macroporous resin chromatography column.

[0187] 2. Preparation of the composition

[0188] T1. 19 parts of Myristicae Fructus were mixed with 100 parts of cyclohexane, heated to reflux for 8 hours, filtered to obtain the filtrate, and dried at 50°C to obtain the Myristicae Fructus extract.

[0189] T2. 15 parts of Polygalae Radix, 7 parts of Morus alba L. and 5 parts of Glycyrrhizae Radix et Rhizoma praeparata cum melle were vacuum-dried at 45°C for 12 hours, crushed by a pulverizer to pass through a 400-mesh sieve, and mixed with 8 times the mass of water to decoct for 5 hours to obtain the filtrate. The residue was mixed with 5 times the mass of water to decoct for 4 hours to obtain the filtrate. Further, the residue was mixed with 3 times the mass of water to decoct for 3 hours to obtain the filtrate. The filtrates obtained by the three decoctions were combined to obtain the total filtrate.

[0190] T3. The Chinese medicine extract obtained in step S4 in 1. Preparation of Chinese medicine extract was mixed with the Myristicae Fructus extract obtained in step T1 and the total filtrate obtained in step T2, and concentrated to a relative density of 1.4 to obtain a clear paste, which was the composition.

[0191] Effect Example 1

[0192] Kidney tonifying effect experiment

[0193] The kidney tonifying effect of the composition provided by each embodiment and comparative example of the present application was evaluated according to the method described in “4.1 Kidney tonifying experiment” in the experimental example of the reference patent CN103405617B.

[0194] 1. Experimental method

[0195] Male SD rats with body weight of 120-150 g were taken, and 12.5% urethane 0.6 mL per rat was injected to make them enter light anesthesia, fixed in back position, and the scrotal skin was sterilized and the bilateral testes were removed. After the operation, 20000 U / kg body weight of penicillin sodium was injected intramuscularly to prevent infection for 3 days. After 3 days, the rats were randomly divided into groups, with five rats in each group. Specifically, the dosages and treatment methods of each group were as follows:

[0196] ①Untreated group: 5 healthy male SD rats with testes not removed, with body weight of 120-150 g. Normal saline was administered by gavage every day.

[0197] ②Model group: SD rats with testes removed by the above method, and normal saline was administered by gavage every day.

[0198] ③Testosterone propionate group: SD rats with testes removed by the above method, and 2 mg / kg of testosterone propionate was subcutaneously injected every day.

[0199] ④Each example and comparative example group: The compositions prepared in each of Examples 1-5 and Comparative Examples 1-7 were appropriately diluted, and each rat was administered by gavage with 1 g / kg body weight of crude drug per day.

[0200] Note: The water content of fresh Cistanche used in each example and comparative example of the present application is as high as 70%-85%, so the calculation formula of the crude drug amount is as follows:

[0201] Crude drug amount (g) = fresh Cistanche amount (g) x 20% + other raw drug amount (g)

[0202] After the above groups were continuously administered by gavage for 30 days, 24 h after the last administration, each group of rats was sacrificed and dissected, and the mass of the preputial gland, seminal vesicle gland, prostate, and levator ani muscle was immediately weighed, and the organ coefficients were calculated, and the results are shown in the following table:

[0203]

[0204] Note: 1, the experimental results are mean ± standard deviation; 2, * represents a significant difference <0.05 compared with the model group, ** represents a significant difference <0.01 compared with the model group, and the test method is t test.

[0205] From the experimental results described in the above table, it can be seen that the compositions of Examples 1-5 have the function of significantly improving the organ coefficients of the preputial gland, seminal vesicle gland + prostate, and levator ani muscle of rats.

[0206] Comparative Example 1 uses quartz sand instead of granular sodium chloride in the preparation of the traditional Chinese medicine extract. Quartz sand cannot increase the salinity of the mixture, thereby reducing the enzyme activity of the plants to protect the active ingredients in fresh Cistanche, and only acts as a grinding aid. Comparative Example 2 uses ultra-fine sodium chloride powder, which can increase the salinity of the mixture, but cannot provide the grinding aid effect of the granular sodium chloride (2-2.8 mm) in Examples 1-5. Comparative Example 3 uses too little sodium chloride, which cannot provide sufficient salinity and grinding aid. The experimental results show that the composition of the traditional Chinese medicine extract prepared by the method of Comparative Examples 1-3 does not have the function of improving the organ coefficients of the rat preputial gland, seminal vesicle + prostate, and levator ani muscle, which is not as good as the composition provided in Examples 1-5.

[0207] Comparative Example 4 does not use nitrogen protection but is exposed to air during the preparation of Cistanche, and the prepared traditional Chinese medicine extract is oxidized by air. The experimental results show that the composition of the traditional Chinese medicine extract prepared by the method of Comparative Example 4 does not have the function of improving the organ coefficients of the rat preputial gland, seminal vesicle + prostate, and levator ani muscle, which is not as good as the composition provided in Examples 1-5.

[0208] Comparative Example 5 uses glacial acetic acid as the acidic substance instead of hydrochloric acid, which cannot achieve sufficient extraction. The experimental results show that the composition of the traditional Chinese medicine extract prepared by the method of Comparative Example 5 does not have the function of improving the organ coefficients of the rat preputial gland, seminal vesicle + prostate, and levator ani muscle, which is not as good as the composition provided in Examples 1-5.

[0209] Comparative Example 6 uses Ophiopogon japonicus instead of Radix Polygalae in the composition, which does not have the function of improving the organ coefficients of the rat preputial gland, seminal vesicle + prostate, and levator ani muscle, which is not as good as the composition provided in Examples 1-5. It can be seen that not all compositions can achieve the kidney-tonifying and kidney-strengthening effect of the composition provided in Examples 1-5.

[0210] Comparative Example 7 uses a conventional water extraction method to extract the traditional Chinese medicine extract. The experimental results show that the composition of the traditional Chinese medicine extract prepared by the method of Comparative Example 7 does not have the function of improving the organ coefficients of the rat preputial gland, seminal vesicle + prostate, and levator ani muscle, which is not as good as the composition provided in Examples 1-5. This shows that not all extraction methods of traditional Chinese medicine extracts can achieve the effect of the traditional Chinese medicine extract provided in Examples 1-5.

[0211] Example 2

[0212] Experiment on improving the sexual function of kidney-yang deficiency male mice

[0213] 1. Establishing a kidney-yang deficiency male mouse model

[0214] Male ICR mice with a body weight of 20-25 g were injected intramuscularly with hydrocortisone at a dose of 25 mg / kg body weight once a day for 14 consecutive days, thereby obtaining kidney-yang deficiency male ICR mice. Ten normal control male ICR mice with a body weight of 20-25 g were injected intramuscularly with 0.9% sodium chloride injection once a day.

[0215] 2. Effects of the composition on the sexual function of kidney-yang deficiency male mice

[0216] The normal control mice (mice injected intramuscularly with 0.9% sodium chloride injection) were given 10 mL / kg body weight of deionized water for gavage. The kidney-yang deficiency male ICR mice were randomly divided into groups, with 10 mice in each group. The kidney-yang deficiency group was given 10 mL / kg body weight of deionized water for gavage; the positive control group was given 1 mg / kg body weight of methyltestosterone tablets for gavage; and the mice in Example 1-5 and Comparative Example 1-7 were given the corresponding composition or Chinese medicine granules (Comparative Example 3) for gavage at a dose of 1 g / kg body weight of crude drug. The gavage was performed once a day for 20 consecutive days, and mating ability detection was performed after the last administration.

[0217] Note: The fresh Cistanche used in each example and comparative example of the present application has a water content of 70%-85%, and therefore the calculation formula of the crude drug amount is as follows:

[0218] Crude drug amount (g) = fresh Cistanche amount (g) × 20% + other raw material drug amount (g)

[0219] Take 18-22 g female ICR mice, and give each female mouse 0.4 mg / kg of diethylstilbestrol suspension for gavage once a day for 3 days before the mating experiment. Synchronize the estrus time with the mating time. After the last administration, select female mice in the estrus stage for the experiment. Perform the mating experiment at night, put one male mouse into each cage for 5 min of adaptation, and then put one female mouse into each cage. Record the video for 20 min using an infrared camera to measure the capture latency (the time from when the female mouse is put into the cage to the first capture of the male mouse), the capture frequency (the number of times the male mouse captures the female mouse within 20 min after the female mouse is put into the cage), and the capture rate (the proportion of male mice in the group that exhibit the capture behavior to the total number n of male mice in the group).

[0220] Among them, the average capture frequency = total capture frequency within 20 min ÷ the number of male mice that exhibit the capture behavior. The experimental results are shown in the following table:

[0221]

[0222] Note: 1. The capture latency experimental results are the average value ± standard deviation; 2. * represents a significant difference <0.05 compared with the kidney-yang deficiency group, and the test method is t-test.

[0223] From the experimental results of the above table, it can be seen that the compositions provided by Examples 1-5 can significantly reduce the capture latency of mice, increase the capture rate and the average capture times, and effectively improve the sexual function of male mice.

[0224] Finally, it should be noted that the above content is only used to illustrate the technical solutions of the present application, and is not a limitation on the scope of protection of the present application. Simple modifications or equivalent replacements of the technical solutions of the present application made by those skilled in the art do not deviate from the essence and scope of the technical solutions of the present application.

Claims

1. A composition for invigorating kidney, characterized in that, The invention is prepared from kidney-strengthening Chinese medicine extract, 14-23 parts by weight of nutmeg, 10-20 parts by weight of polygala tenuifolia, 5-9 parts by weight of mulberry fruit, and 3-8 parts by weight of roasted liquorice root; The preparation method of the kidney-strengthening Chinese medicine extract comprises the following steps: S1. Grinding and sieving the Tribulus terrestris to obtain Tribulus terrestris powder; S2. Take fresh Cistanche deserticola and sodium chloride, mash and grind them together, heat under nitrogen protection, then add methanol, concentrated hydrochloric acid, and the Tribulus terrestris powder obtained in step S1, reflux, and filter to obtain a crude extract; S3, concentrating the crude extract obtained in step S2, adjusting the pH, and obtaining a concentrated solution; S4, separating the concentrated solution obtained in step S3 by column chromatography, concentrating and drying the eluate to obtain a kidney-strengthening Chinese medicine extract; In parts by mass, the amount of Tribulus terrestris in step S1 is 23-32 parts; the amount of fresh Cistanche deserticola in step S2 is 100-120 parts; in step S2, the amount of sodium chloride is 45-55 parts, the amount of methanol is 150-250 parts, the amount of concentrated hydrochloric acid is 0.8-1.5 parts, and the sodium chloride in step S2 is granular sodium chloride.

2. The composition of claim 1, wherein, The grinding time in step S2 is 10-45 minutes, and the heating temperature under nitrogen protection is 65-85° C. and the heating time is 20-50 minutes.

3. Process for the preparation of a composition according to any one of claim 1 or claim 2, characterized in that, The following steps are involved: T1. Mixing nutmeg with an organic solvent, heating under reflux, filtering the filtrate, concentrating, and drying to obtain a nutmeg extract; T2. Dry and decoct the Polygala tenuifolia, Morus alba, and Radix Glycyrrhizae, filter, and combine the filtrates to obtain the total filtrate; T3. The kidney-strengthening Chinese herbal extract, the nutmeg extract obtained in step T1, and the total filtrate obtained in step T2 are mixed and concentrated to obtain a composition.

4. The production method according to claim 3, characterized by, In step T1, the organic solvent is selected from at least one of n-hexane, cyclohexane, acetone, and ethanol.

5. A kidney tonic medicine, characterized in that, The raw material is the composition described in any one of claims 1-2 or the composition prepared by the preparation method described in any one of claims 3-4.

6. Use of the composition according to any one of claims 1-2, the preparation method according to any one of claims 3-4, or the kidney-tonifying drug according to claim 5 in the production of kidney-tonifying and kidney-strengthening products.

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