CN112957391A - Thymus plant extract, preparation method and application - Google Patents

Thymus plant extract, preparation method and application Download PDF

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CN112957391A
CN112957391A CN202110186091.2A CN202110186091A CN112957391A CN 112957391 A CN112957391 A CN 112957391A CN 202110186091 A CN202110186091 A CN 202110186091A CN 112957391 A CN112957391 A CN 112957391A
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thyme
thymus
plant
extract
liver
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折改梅
闫昕
王秀环
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Beijing University of Chinese Medicine
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/51—Concentration or drying of the extract, e.g. Lyophilisation, freeze-drying or spray-drying
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

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Abstract

The invention relates to a thymus plant extract, a preparation method and application thereof. The invention is based on the discovery that thymus plants have good curative effect on liver injury by the inventor, and the invention is provided. Meanwhile, the liver protection effect of the total extract of the thymus plant is obviously better than that of different component types and parts, and the liver protection activity of the thymus plant is proved to have a multi-component synergistic effect. The medicine disclosed by the invention is high in safety, readily available in raw materials, remarkable in curative effect and wide in application prospect in the field of liver injury prevention and treatment.

Description

Thymus plant extract, preparation method and application
Technical Field
The invention relates to the technical field of traditional Chinese medicine product research, and particularly relates to an extract, a preparation method and application.
Background
The liver is an important parenchymal metabolic organ of a human body, is also the largest digestive gland in the digestive system, and plays an important role in the aspects of digestion, detoxification, metabolism, immunoregulation, hematopoiesis and the like of the body. Liver injury is a common liver disease, including hepatitis, fatty liver, liver fibrosis, cirrhosis and liver cancer, and is one of the major health problems of humans.
The causes of liver injury are various and mainly classified into drug-induced liver injury, chemical liver injury, non-alcoholic fatty liver injury and immunological liver injury. The drug-induced liver injury refers to liver injury induced by various prescription or non-prescription chemical drugs, biological agents, traditional Chinese medicines, natural medicines, health products, dietary supplements, metabolites thereof, auxiliary materials and the like; chemical liver injury is liver injury caused by alcohol, chemical toxic substances in the environment and other hepatotoxic substances; non-alcoholic fatty liver injury refers to the clinical pathological syndrome characterized mainly by diffuse hepatocellular steatosis, except for alcohol and other definite liver damage factors; immune liver injury is an autoimmune liver disease based on immune response and mainly expressed by inflammatory cell infiltration of liver tissues and hepatocyte necrosis.
In the syndrome of traditional Chinese medicine, liver injury can be classified into jaundice and hypochondriac pain according to its clinical manifestations. The traditional Chinese medicine researches on etiology and pathogenesis of liver injury: non-alcoholic liver injury is mainly caused by improper diet, emotional disorder, fatigue and loss of tolerance and constitutional factors. Liver depression and qi stagnation, spleen dysfunction in transportation and transportation, qi and blood phlegm stasis in hypochondrium, obstruction of liver meridian are basic pathogenesis; alcoholic liver injury is the condition of excessive drinking, retention of damp-heat toxin in the interior, liver damage and spleen damage, liver catharsis and spleen transportation and transformation failure, disharmony of qi and blood, phlegm turbidity generation, qi and blood phlegm-dampness mutual pulsation, and stagnation under the hypochondrium to form lumps. The long-term persistence can cause the dysfunction of the liver, the spleen and the kidney, and the qi and the blood are combined to form the swelling; the direct disease of drug-induced liver injury is mainly caused by the pathogenic factors of drug-induced toxicity, and the key pathogenesis is liver dysfunction and diarrhea, and spleen dysfunction.
Liver damage is a serious threat to human health, and the World Health Organization (WHO) survey reports show that the population suffering from various degrees of liver disease worldwide is about 10%. The search for liver-protecting drugs is ongoing, in recent years, natural plant ingredients have attracted attention in this field, and more researchers have begun to explore the possibility of natural medicinal plants and their active ingredients in the prevention and treatment of liver injury.
Thyme (A)Thymus vulgaris.Linn Aeus) is Thymus of Labiatae (Labiatae) ((R)Thymus) The plant is known as perennial low bush. The plants are named because of strong aromatic odor in the flowering period. According to record of book 66 (1977) of Chinese plant record, 300-400 thyme species exist in the world, the origin is the coast of Mediterranean sea, and the thyme species are widely distributed in Europe, North America and Asia at present. According to the related information, the Thymus chinensis has 17 varieties and 2 varieties including Thymus vulgaris: (A), (B), (CT. mongolicusRonn.), and thyme (a) from a different plant (A. serpyllum)T. marschalianusWilld.), thyme short (thyme(s) ((s))T. mandschuricus Ronn.)、Thymus serpyllum (L.) MoenchT. disjunctusKlok.), thyme (xing Kai), (xi Shi xi), (xi Kai), (xi Shi xi) (Klok.) (xi Shi xi), (xi Shi xi) (thyme (xi) (xiT. przewalskiiKom.), thyme (xing' an: (A. am.)T. dahuricusSerg.), thyme (thyme) and (thyme)T. proximusShrg.), thyme (thyme) of lineal leaf (T. linearisBenth.) and Altha thyme (T. altaicusKlok. et Shost.), thyme (thyme) oblique (T. inaequalisKlok.), thyme (thyme for developing pulse: (A)T. nervulosusKlok.), thyme of black dragon river (T. amurensisKlok.), thyme brevifolia (thyme gracile (L.), (T. curtusKlok.), thyme from alpine (thyme: (a) ((b))T. diminulusKlok.), thyme rose (T. roseusSchipcz.), thyme (Zanthoxylum thyme: (A. am. F.)T. seravschanicusKlok.), red pepper (T.quinquecostatusCelak.) these 17 species and the wild pepper variety Zhanmai (T.quinquecostatus Celak. var. przewalskii) Asian variety (Asian sweet pepper) ((Asian sweet pepper))T.quinquecostatus Celak. var. asiaticus) These 2 variants. The gravel slope land and the grassland in arid and semiarid mountain areas are mainly distributed in Xinjiang, Gansu, Qinghai, Ningxia, Shaanxi, inner Mongolia, Shanxi, Shandong, Anhui, Hebei, Liaoning, Heilongjiang and other lands in the north of the yellow river.
The chemical composition of thyme can be divided into two major categories, volatile oil and non-volatile oil. The volatile oils are mainly composed of terpenes, esters, phenolic derivatives, and the like. The nonvolatile oil mainly comprises flavone, phenylpropanoid, tannin, organic acid, terpenes and phytosterol. Research shows that thyme plant has wide pharmacological activity including antibiosis, antioxidation, antitumor, antiphlogosis, pain relieving, cough relieving, blood pressure lowering, blood sugar lowering, parasite expelling, etc. The main chemical component types of thymus plants suggest that they may have the potential to protect the liver. At present, few research reports on the liver protection function of thymus plants exist, application examples exist, and systematic research and analysis are lacked. The invention researches the liver protection activity of the thymus plant of China.
Disclosure of Invention
The invention provides a traditional Chinese medicine extract which is extracted from Chinese thymus plants, and the preferred thymus plants comprise thyme, thyme of different strains, thyme of short section, thyme of long teeth, thyme of Xingkai, thyme of Xingan, thyme of pseudo-thyme, thyme of linear leaves, thyme of altai, thyme of oblique leaves, thyme of pulse display, thyme of Heilongjiang, thyme of short hair, thyme of Gaoshan, thyme of rose, thyme of black-pepper, thyme of thym.
The invention also provides a preparation method of the thymus plant extract, which comprises the following steps:
(1) taking 100 parts by weight of thymus plants including thyme, short thyme, altai thyme, black dragon river thyme, long teeth thyme, rose thyme, xingan thyme, leafy thyme, alpine thyme, Xingkai thyme, linear thyme, allophytic thyme, pulse-developing thyme, short thyme, pseudo thyme, thyme variant of thyme or thyme Asian thyme, adding ethanol, heating, refluxing and extracting, and repeatedly extracting for several times to completely extract;
(2) mixing the materials obtained by the several times of extraction, recovering the solvent, and drying under reduced pressure to obtain an extract dry extract.
Preferably, the extraction method comprises the following material-liquid ratio: 1: 20-1: 80 (g: mL).
Preferably, the extraction time of each extraction method is 40-120 minutes.
Preferably, the concentration of the solvent ethanol in the extraction method is 50-90%.
Preferably, the extraction times of the extraction method are 2-4 times.
Preferably, the solvent recovery process temperature is: 30 to 70 ℃.
The thyme plant of the present invention can be taken from different places, for example, from the arid and semiarid mountain gravel slope and grassland of Xinjiang, Gansu, Qinghai, Ningxia, Shaanxi, inner Mongolia, Shanxi, Shandong, Anhui, Hebei, Liaoning, Heilongjiang and other places in the north of the yellow river.
The invention also provides the application of the thymus plant or the traditional Chinese medicine extract prepared by the preparation method in preparing the medicine for preventing and treating acute and chronic liver injury.
The invention also provides a traditional Chinese medicine extract which is prepared by the preparation method.
The invention provides a thymus plant extract, a preparation method and application thereof, and the extract has obvious liver protection activity and no toxic or side effect.
The thymus plant extract can be produced by using a conventional method in the existing pharmaceutical field according to actual needs, and prepared into any clinically or pharmaceutically acceptable dosage form to be applied to a patient needing treatment in an oral or parenteral administration mode, such as: such as powder, granule, decoction, tablet, dispersible tablet, sustained release tablet, sublingual tablet, orally disintegrating tablet, capsule, soft capsule, microcapsule, pill, dripping pill, concentrated pill, pellet, syrup, aerosol, injection, oral liquid, gel, microsphere, sustained release preparation, controlled release preparation, quick release preparation, etc. The preferable dosage forms are powder, granule, and capsule.
In the process of preparing the above-mentioned all dosage forms, any one or more excipients or carriers, such as absorbents, inclusion agents, diluents, wetting agents, binders, disintegrants, retarding agents, lubricants, glidants, sweeteners, coating agents, clarifiers, preservatives, mucilage agents, buffering agents, solubilizers, antioxidants, solubilizing agents, aromatizers, bases, etc., may be appropriately added according to actual requirements by using conventional methods in the pharmaceutical field.
The selective range of the absorbent comprises: active carbon, calcium sulfate, calcium hydrophosphate, calcium carbonate, magnesium oxide, magnesium carbonate and aluminum hydroxide gel powder.
The selection range of the inclusion agent comprises:  -cyclodextrin, beta-cyclodextrin, hydroxypropyl-beta-cyclodextrin.
The diluent selection ranges include: dextrin, water soluble dextrin, spray dried lactose, starch, soluble starch, compressible starch, lactose, mannitol, chitosan, xylitol, bifidobacterium, fast flowing lactose, anhydrous beta-lactose,  -lactose-hydrate, and anhydrous  -lactose.
The wetting agent selection ranges include: water and ethanol.
The adhesive selection ranges include: starch slurry, dextrin slurry, ethanol syrup (ethanol concentration less than 60%), sucrose syrup, liquid glucose, maltose, refined honey, acacia slurry, gelatin slurry, compressible starch, dextrin, polyvinylpyrrolidone, sodium carboxymethylcellulose, low-substituted hydroxypropyl cellulose, hydroxypropyl methylcellulose, microcrystalline cellulose, and polyethylene glycol.
The selection range of the disintegrating agent comprises: sodium carboxymethyl starch, dried starch, carboxymethyl starch, microcrystalline cellulose, konjac powder, magnesium trisilicate, tween-80, sodium lauryl sulfate, sodium stearyl sulfonate, tween-80, hydroxypropyl starch propionate, croscarmellose sodium, M CC, HPM C, L-HPC, Avicel PH101, Avicel PH102, Avicel PH103, Avicel PH105, Avicel PH301, Avicel PH302, Avicel RC591, Emcocel.
The selection range of the retarder comprises: PVP, polyethylene glycol and benzene two formic acid cellulose.
The lubricant selection ranges include: magnesium stearate, talcum powder, polyethylene glycol, liquid paraffin, water-soluble sodium lauryl sulfate and boric acid.
The selection range of the glidant comprises: silica gel, aluminum hydroxide gel powder and fast flow lactose.
The sweetener selection range comprises: stevia sugar, high fructose, i.e., aspartame, xylitol, sodium cyclamate,  -sorbitol, beta-sorbitol, gamma-sorbitol, sucrose, saccharin sodium, simple syrup, and aromatic syrup.
The selection range of the coating agent comprises: HPM C, PVP, polyethylene glycol, acrylic resins, aqueous acrylic resin dispersions, ethylcellulose pseudolatex, cellulose acetate phthalate latex, aqueous hydroxypropyl methylcellulose acetate succinate dispersion, methylcellulose, hydroxypropyl cellulose, aqueous polyvinyl acetate phthalate dispersion latex, carboxymethyl ethyl cellulose, shellac, hydrogenated vegetable oils, poly (ethyl acetate-methacrylic acid acrylate), poly (ethyl acetate-trimethyl ammonium methacrylate acetate) pseudolatex, chitin and chitosan, low-substituted hydroxypropyl cellulose, acrylic resin coating materials I, II, III, iv and sodium carboxymethyl cellulose, compressible starch, sucrose fatty acid esters, ethyl cellulose.
The range of fragrance selections includes: lemon, peppermint oil, apple essence, banana essence flavor agent, menthol, liquorice sweet essence and sodium cyclamate.
Antioxidant selection ranges include: dibutylhydroxytoluene, gallic acid, ascorbic acid, sodium ascorbate and malic acid.
The selection range of the mucilage comprises: mucilage such as sodium alginate, Arabic gum, gelatin, methyl cellulose, ethyl maltol, steviosin, sodium carboxymethyl cellulose and the like.
Buffer selection ranges include: tartaric acid, citric acid and sodium bicarbonate.
The range of solubilizer choices includes: tween 80, poloxamer, span and soybean lecithin.
Clarifying agent selection ranges include: gelatin, 101 juice clarifying agent and ZTC 1 + 1 natural clarifying agent.
The selection range of the cosolvent comprises: ethanol, ethylene glycol, polyethylene glycol, propylene glycol, glycerol.
Preservative selection ranges include: parabens, benzoic acid and its salt, parabens, methyl paraben, propyl paraben, sorbic acid and its salt, chlorhexidine acetate, and domiphen bromide.
The matrix selection range comprises: water-soluble matrix (polyethylene glycol, PEG-4000/6000, sodium stearate, glycerol, gelatin, water, etc.), fat-soluble matrix (selected from beeswax, stearic acid, hydrogenated vegetable oil, etc.), and delayed release matrix (stearic acid, glyceryl monostearate, insect wax, beeswax, hydrogenated oil and vegetable oil, polyoxyethylene monostearate (S-40), stearyl alcohol, semisynthetic fatty acid ester, polyethylene glycol, surfactant, etc.).
The preparation of the powder of the invention comprises the following steps: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Taking 1 part of dry extract powder and 1.3 parts of dextrin and lactose (2: 1), and fully and uniformly mixing to obtain the traditional Chinese medicine.
Preparation of the granules of the invention: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Taking 1 part of dry extract powder, 1.3 parts of dextrin and lactose (2: 1), fully and uniformly mixing, granulating by using 80% ethanol, drying and finishing granules to obtain the traditional Chinese medicine composition.
The preparation of the capsule of the invention: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Taking 1 part of dry extract powder, 1.3 parts of dextrin and lactose (2: 1), fully and uniformly mixing, granulating by using 80% ethanol, drying, finishing granules, and filling the granules into capsules to obtain the capsule.
The preparation of the soft capsule of the invention comprises the following steps: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Adding gelatin into a reaction tank, adding appropriate amount of water under stirring, sealing, adding glycerol after gelatin is completely dissolved, stirring, vacuum degassing for 2 hr, placing into a heat-insulating barrel, and standing overnight. Mixing vegetable oil, lecithin and insect wax, adding dry extract powder, stirring, grinding to colloid, and standing at room temperature. Pouring the medicinal liquid into a soft capsule machine, making into soft capsule, and drying.
Preparation of the tablets of the invention: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Mixing dry extract powder 1 part, dextrin and lactose (2: 1) 1.3 parts, granulating with 80% ethanol, drying, grading, and tabletting.
The preparation of the sustained-release tablet of the invention comprises the following steps: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Taking 1 part of dry extract powder, 1.3 parts of dextrin and lactose (2: 1), fully and uniformly mixing, granulating by using 80% ethanol, granulating by using a 18-mesh sieve, drying at 55 ℃, granulating by using a 16-mesh sieve, adding sodium carboxymethyl starch, uniformly mixing, and pressing by using a tablet press to obtain a tablet core; then preparing a slow-release coating mixed solution, and uniformly grinding the acrylic acid II resin, HPMC, PEG-6000, tween-80, castor oil, talcum powder, titanium dioxide and magnesium stearate by a colloid mill. Finally, the tablet cores are placed in a sugar coating pan, sprayed with liquid by a spray gun for coating, polished, cooled and dried for about 1 hour at 50 ℃ to obtain the tablet cores.
The preparation of the sublingual tablet of the invention comprises the following steps: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Taking 1 part of dry extract powder, hydroxypropyl methylcellulose, microcrystalline cellulose and talcum powder, uniformly mixing by an equivalent incremental method, and pressing by a tablet press to obtain the traditional Chinese medicine.
The preparation of the pill of the invention: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 120-mesh sieve to obtain dry extract powder. Sieving the film forming powder with a 120-140-mesh sieve, placing the film forming powder into a sugar coating pot, spraying a proper amount of clear water, starting a machine, uniformly moistening the film forming powder until the film forming powder is held by a hand to form a mass, dispersing the mass by light pressure, and granulating. Putting the granules into a sugar coating pan, slightly adding a little dry powder, starting the pan, rotating to spherical shape, taking out, sieving and the like. Then the spherical film is put into a sugar-coating pan, and powder is evenly added to spray water, so that the pills are evenly enlarged. Selecting uniform and qualified pills, covering with medicinal fine powder (100-120 times), rolling in a sugar-coating pan to make the pills round and compact, screening the pills, and oven-drying in an oven at 60-80 deg.C.
The preparation of the dripping pill of the invention: taking dry extracts of thymus plant thymus mongolicus, thyme with different strains, thyme with short sections, thyme with long teeth, thyme with Xingkai, thyme with Xingan, thyme with pseudo-thyme, thyme with linear leaves, thyme with altai, thyme with oblique leaves, thyme with pulse-appearing, thyme with black dragon river, thyme with short hair, thyme with alpine, thyme with rose, thyme with black-pepper, thyme variant with thyme with spread hair or thyme with Asia, crushing and sieving with a 80-mesh sieve to obtain dry extract powder. Taking PEG 4000: PEG 6000 is 1:2, dry extract powder: mixing the ground substances at a ratio of 1:2, placing in a container, heating to 70-80 deg.C, cooling to 40-50 deg.C, dropping into dripping pill machine, dropping into liquid paraffin or dimethyl silicone oil at 6-8 deg.C, making into dripping pill, and removing oil.
Compared with the prior art, the invention has the following advantages and effects:
1. the inventor systematically discovers that the thymus plant has good prevention and treatment effects on liver injury for the first time.
2. The thyme plant has wide medicinal and edible values, so the raw materials are easy to obtain and have high safety, and the thyme plant has wide application prospect in the field of prevention and treatment of liver injury diseases.
Drawings
FIG. 1 is a HE staining pattern (X100) of pathological sections of liver tissues of mice in each experimental group.
Detailed Description
In order that the technical solutions of the present invention may be better understood and applied, the present invention will be further described and illustrated in detail with reference to the following specific examples, but the embodiments of the present invention are not limited thereto.
Example 1 preparation of thymus extract method one
1) Taking thymus plants including thyme, short-section thyme, Altai thyme, black-QI thyme, black dragon river thyme, long-teeth thyme, rose thyme, Xingan thyme, leafy thyme, alpine thyme, Xingkai thyme, linea thyme, allophytic thyme, pulse-showing thyme, short-hair thyme, pseudo-thyme, thyme variant or Asian thyme 100g, adding 6L of 50% ethanol according to the material-liquid ratio of 1:60 (g: mL), heating and refluxing for extraction for 60 minutes to obtain decoction and decoction dregs.
2) Extracting the residue for 2 times according to the extraction operation in step 1). Mixing the extractive solutions obtained in step 1) and the above repeated extraction, filtering, concentrating, and drying to obtain corresponding plant extract.
Example 2 preparation of Thymus extract
1) Taking thymus plants including thyme, short-section thyme, Altai thyme, black-QI thyme, black dragon river thyme, long-teeth thyme, rose thyme, Xingan thyme, leafy thyme, alpine thyme, Xingkai thyme, linea thyme, allophytic thyme, pulse-showing thyme, short-hair thyme, pseudo-thyme, thyme variant or Asian thyme 100g, adding 4L of 80% ethanol according to the material-liquid ratio of 1:40 (g: mL), heating and refluxing for extraction for 80 minutes to obtain decoction and decoction dregs.
2) Extracting the residue for 1 time according to the extraction operation in step 1). Mixing the extractive solutions obtained in step 1) and the above repeated extraction, filtering, concentrating, and drying to obtain corresponding plant extract.
Example 3 preparation of Thymus extract
1) Taking thymus plants including thyme, short-section thyme, Altai thyme, black-QI thyme, black dragon river thyme, long-teeth thyme, rose thyme, Xingan thyme, leafy thyme, alpine thyme, Xingkai thyme, linea thyme, allophytic thyme, pulse-showing thyme, short-hair thyme, pseudo-thyme, thyme variant or Asian thyme 100g, adding 70% ethanol 2L according to the material-liquid ratio of 1:20 (g: mL), heating and refluxing for extraction for 80 minutes to obtain decoction and decoction dregs.
2) Extracting the residue for 2 times according to the extraction operation in step 1). Mixing the extractive solutions obtained in step 1) and the above repeated extraction, filtering, concentrating, and drying to obtain corresponding plant extract.
Example 4 preparation of Thymus extract
1) Taking thymus plants including thyme, short-section thyme, Altai thyme, black-QI thyme, black dragon river thyme, long-teeth thyme, rose thyme, Xingan thyme, leafy thyme, alpine thyme, Xingkai thyme, linea thyme, allophytic thyme, pulse-showing thyme, short-hair thyme, pseudo-thyme, thyme variant or Asian thyme 100g, adding 65% ethanol 4L according to the material-liquid ratio of 1:40 (g: mL), heating and refluxing for extraction for 120 minutes to obtain decoction and decoction dregs.
2) Extracting the residue for 1 time according to the extraction operation in step 1). Mixing the extractive solutions obtained in step 1) and the above repeated extraction, filtering, concentrating, and drying to obtain corresponding plant extract.
Example 5 Thymus extract hepatoprotective effect test
The invention adopts Lieber-DeCarli alcohol liquid feed to feed and induces a mouse liver injury model. The protective effect of thymus plant extract on liver injury was evaluated using alanine Aminotransferase (ALT), aspartate Aminotransferase (AST), Total Cholesterol (TC), and total Triglyceride (TG) as indices.
1. Experimental Material
(1) Laboratory animal
220 SPF male C57/6N mice, weighing 20.0 + -2.0 g, were provided by the Beijing Wintolite laboratory animal technology, Inc. (license number SYXK (Jing) 2017-.
(2) Medicaments and agents
The Lieber-DeCarli alcohol liquid feed is provided by southbound telofil feed science and technology ltd; the serum biochemical index (AST, ALT, TC, TG) detection kit is provided by Beckman company in Germany.
2. Experimental methods
(1) Preparation of thymus plant extract
1) Taking 100g of thyme plant, adding 4L of 80% ethanol according to the material-liquid ratio of 1:40 (g: mL), heating and refluxing for extraction for 80 minutes to obtain decoction and dregs.
2) Extracting the residue for 1 time according to the extraction operation in step 1). Mixing the extracting solutions obtained in the step 1) and the repeated extraction, filtering, concentrating and drying to obtain dry extractum of the thymus plant extract.
(2) Grouping animals
The mice of C57/6N were bred and bred adaptively in an SPF-level environment for one week, and then divided into a normal group, a model group, a silymarin treatment group, a thyme pup treatment group, an Altai thyme treatment group, a thyme treatment group of black-QIAO thyme, a thyme treatment group of black Longjiang thyme, a thyme treatment group of long teeth thyme, a thyme treatment group of rose thyme, a thyme treatment group of Xingan thyme, a thyme treatment group of oblique leaf thyme, a thyme treatment group of Gaoshan thyme, a thyme treatment group of Xingkai thyme, a thyme treatment group of line leaf thyme, a thyme treatment group of abnormal plants, a thyme treatment group of pulse-showing thyme, a thyme treatment group of short hair, a thyme-like treatment group, a thyme treatment group, a.
(3) Animal modeling and administration
A liver injury model of a mouse is established by adopting a Lieber-Decalli liquid feed pairing feeding method. The normal group was fed with control liquid diet (no alcohol), and the model group and each thyme treatment group were fed with alcohol liquid diet (4% V/V) for 6 weeks. During the period, the thymus plant treatment group is respectively administrated with 120 mg/kg of corresponding plant extract by intragastric administration daily, and the silymarin treatment group is administrated with 60 mg/kg of silymarin.
(4) Detection method
1) General observations
After the completion of 6 weeks of feeding, the general conditions of the mice, including food intake, body hair, mental state, activity and body weight, were observed.
2) Serum biochemical index detection
After 6 weeks of experiment, the mice in each group are anesthetized by chloral hydrate, blood is taken and placed in a centrifuge tube, centrifugation is carried out at 4000 r/min for 15min, and serum is separated. ALT, AST, TC and TG levels in serum are detected by a Beckmann full-automatic biochemical analyzer.
3. Results
(1) General observations
The body weight and the food intake of each group of mice in the experimental period have no obvious difference; the mice in the normal group are normal in performance, good in mental state and stable in behavior, and have no drunkenness such as lethargy, walking instability and standing instability. The model group mice show intoxication manifestations such as lethargy, slow movement, slow response, bad mental state, unstable walking and standing. The above-mentioned adverse symptoms were improved in each of the thymus-treated and silymarin-treated mice. The skin of the mice in the normal group, the thymus plant treatment group and the silymarin treatment group is glossy, while the skin of the mice in the model group is loose and lacks luster, and has a depilation phenomenon.
(2) Serum biochemical index detection result
As shown in Table 1, compared with the normal group of mice, the serum levels of ALT, AST, TC and TG of the model group of mice are all obviously increased (P is less than 0.01), which indicates that the liver has abnormal functions and cell damage, and the liver damage model is successfully established. Compared with the model group, the levels of the biochemical indexes in the serum of the thymus plant treatment group and the silymarin treatment group are remarkably reduced (P is less than 0.05 or P is less than 0.01).
TABLE 1 influence of Thymus extracts on Biochemical indices of mouse serum (` X. + -. S)
Group of ALT (U/L) AST (U/L) TC (mmol/L) TG (mmol/L)
Normal group 32.00±5.14 130.20±7.39 3.48±0.38 2.17±0.21
Model set 74.00±15.99## 181.12±18.43## 4.08±0.21## 2.82±0.37##
Silymarin treatment group 43.90±6.89## 152.94±7.76## 3.52±0.32## 2.24±0.32##
Thyme treatment group 44.72±7.68** 148.16±12.29** 3.64±0.34* 2.40±0.26*
Short thyme treatment group 46.34±8.70** 143.56±10.63** 3.59±0.31* 2.35±0.53*
Altai thyme treatment group 57.64±17.47** 157.47±23.79** 3.64±0.55* 2.41±0.39*
Thyme treatment group of Wuqia 51.61±10.47** 158.48±18.78** 3.59±0.73* 2.37±0.48*
Heilongjiang thyme treatment group 41.10±5.86** 143.53±17.54** 3.62±0.49* 2.40±0.38*
Long-tooth thyme treatment group 47.30±8.98** 150.96±12.00** 3.50±0.47** 2.34±0.37*
Rose and thyme treatment group 47.00±13.91** 150.66±11.75** 3.10±0.65** 2.26±0.27**
Xingan thyme treatment group 49.40±22.87** 143.13±13.06** 3.58±0.40** 2.34±0.29*
Thymus serpyllum treatment group 45.70±13.05** 149.23±12.68** 3.70±0.41* 2.41±0.47*
High mountain thyme treatment group 56.00±16.11** 151.90±15.15** 3.24±0.57** 2.36±0.83*
Xingkai thyme treatment group 52.50±11.14** 153.83±11.84** 3.46±0.52** 2.43±0.66*
Thyme herb line leaf treatment group 48.30±9.81** 150.46±9.71** 3.69±0.32* 2.37±0.40*
Special thyme treatment group 40.80±8.92** 144.17±8.05** 3.50±0.36** 2.36±0.39*
Thyme treatment group with pulse showing function 52.60±9.62** 147.13±9.29** 3.69±0.39* 2.27±0.31**
Thymus serpyllum treatment group 54.20±10.21** 167.51±20.78* 3.68±0.37* 2.36±0.63*
Thyme-imitating treatment group 46.30±12.07** 161.59±23.48** 3.56±0.36** 2.26±0.47**
Mongolian thyme herb treatment group 39.50±4.99** 142.14±7.72** 3.58±0.32** 2.38±0.27*
Thymus-spread variant treatment group 48.60±10.42** 142.08±9.04** 3.54±0.34** 2.42±0.31*
Asian thyme herb treatment group 45.30±13.59** 154.16±17.09** 3.40±0.41** 2.42±0.45**
Note: # P <0.05, # P <0.01 compared to normal; comparison with model group of P <0.05, P <0.01
4. Conclusion
The research result shows that the thymus plant extract can improve the rise of serum transaminase, total cholesterol and total triglyceride caused by liver injury and has a protective effect on liver function.
Example 6 liver protection histopathology Studies of Thymus plant extracts
1. Experimental Material
(1) Laboratory animal
SPF-grade male C57/6N mice 210, weighing 20.0 + -2.0 g, were provided by the Beijing Wintolite laboratory animal technology Co., Ltd (license number SYXK (Jing) 2017-.
(2) Medicaments and agents
The Lieber-DeCarli alcohol liquid feed is provided by southbound telofil feed science and technology ltd; paraformaldehyde, hematoxylin, eosin, differentiation solution, xylene, rewet solution, differentiation solution, neutral gum, section paraffin, glycerol, glass slide, and cover slip are provided by seville Biotech limited.
2. Experimental methods
(1) Preparation of thymus plant extract
1) Taking 100g of thyme plant, adding 4L of 80% ethanol according to the material-liquid ratio of 1:40 (g: mL), heating and refluxing for extraction for 80 minutes to obtain decoction and dregs.
2) Extracting the residue for 1 time according to the extraction operation in step 1). Mixing the extracting solutions obtained in the step 1) and the repeated extraction, filtering, concentrating and drying to obtain dry extractum of the thymus plant extract.
(2) Grouping animals
The mice of C57/6N were bred and bred adaptively in an SPF-level environment for one week, and then divided into a normal group, a model group, a thyme treatment group, a short thyme treatment group, an altai thyme treatment group, a black thyme treatment group, a long thyme treatment group, a rose thyme treatment group, a Xingan thyme treatment group, a thyme treatment group of alpine thyme, a Xinkhaki thyme treatment group, a thyme treatment group of linear thyme, an allogenic thyme treatment group, a thyme-developing thyme treatment group, a thyme-short thyme treatment group, a thyme-simulating treatment group, a thyme-pepper variety treatment group, an Asian thyme treatment group, and 10 mice per group.
(3) Animal modeling and administration
A liver injury model of a mouse is established by adopting a Lieber-Decalli liquid feed pairing feeding method. The normal group was fed with control liquid diet (no alcohol), and the model group and each thyme treatment group were fed with alcohol liquid diet (4% V/V) for 6 weeks. During the treatment period, 120 mg/kg of the corresponding plant extract was administered to each thymus treatment group daily.
(4) HE staining of liver tissue sections
1) Preparing a paraffin section: taking a proper amount of liver tissue paraformaldehyde, fixing for 48h, and then dehydrating with gradient alcohol, and making into tissue wax block after transparency and wax penetration. The tissue wax block was cut into 5 μm thick sections with a microtome and mounted on glass slides. Then baking slices, dewaxing and rehydrating for later use.
2) Dyeing: staining tissue sections with hematoxylin staining solution for 1-5 min, washing off excessive staining solution with double distilled water, performing microscopic examination after differentiation and fading of 0.5% hydrochloric acid alcohol solution for 20s, washing with double distilled water for 1 min, returning blue with 1% dilute ammonia water for 30 s, washing with distilled water for 1 min, staining with eosin staining solution for 20s-300s, and washing off excessive staining solution with double distilled water.
3) Dehydration, transparency and mounting: and dehydrating the dyed tissue slices in 85% ethanol solution for 20s, dehydrating in 90% ethanol solution for 30 s, dehydrating in 95% ethanol solution for 1 min twice, dehydrating in absolute ethanol for 2 min twice, and removing excessive ethanol by suction. And then the dehydrated tissue slices are transparent for 2 min by xylene, repeated for 3 times, and finally, the slices are sealed by neutral gum.
4) And (3) observing tissue morphology: and (3) scanning the slices under a super-resolution micro-imager to obtain images, and observing and comparing morphological changes of tissues of each group.
3. Results
As shown in FIG. 1, the hepatocytes of the normal group of mice were arranged radially with the central venous lumen as the center, with clear liver cords, intact hepatocyte structure, clear outline, and no significant swelling or steatosis. The liver of the model group mouse is obviously subjected to steatosis, a large number of fat vacuoles appear in cells, inflammatory cell infiltration is visible in the central veins and the central area of the lobule, the cell outline is fuzzy, and a large number of necroses appear. The volume and the number of fat drops in the liver cells of the mice in each thymus plant treatment group are reduced, the liver cords are arranged regularly, and the cell and tissue structure characteristics are close to those of the mice in a normal group.
4. Conclusion
The research result shows that the thymus plant extract has a protective effect on the liver and can improve the degree of liver injury.
Example 7 synergistic Effect study of Thymus for liver protection
By taking thyme as an example, the anti-liver injury activity of the total extract and the main component types of the thyme plant is compared and researched, and the synergistic effect of the liver protection activity of the thyme plant is clear.
1. Experimental Material
(1) Laboratory animal
SPF-grade male C57/6N mice, weighing 20.0 + -2.0 g, were obtained from Beijing Wintolite laboratory animal technology, Inc. (license number SYXK (Jing) 2017-.
(2) Medicaments and agents
The Lieber-DeCarli alcohol liquid feed is provided by southbound telofil feed science and technology ltd; the serum biochemical index (AST, ALT, TC, TG) detection kit is provided by Beckman company in Germany.
2. Experimental methods
(1) Preparation of total extract of herba Thymi
1) Taking 100g of Japanese pepper, adding 4L of 80% ethanol according to the material-liquid ratio of 1:40 (g: mL), heating and refluxing for extraction for 80 minutes to obtain decoction and dregs.
2) Extracting the residue for 1 time according to the extraction operation in step 1). Mixing the extractive solutions obtained in step 1) and the above repeated extraction, filtering, and concentrating to obtain total extract of herba Zanthoxyli.
(2) Preparation of triterpenoid part of thyme herb
1) Taking 100g of each thyme plant, adding 4L of 80% ethanol according to the material-liquid ratio of 1:40 (g: mL), heating and refluxing for extraction for 80 minutes to obtain decoction and dregs.
2) Extracting the residue for 1 time according to the extraction operation in step 1). Mixing the extractive solutions obtained in step 1) and the above repeated extraction, filtering, concentrating, and drying to obtain dry extract.
3) Taking the dry extract obtained in the step 2), adding water to disperse the dry extract into a solution of 2 g/mL, and loading the solution onto an AB-8 macroporous resin column. Eluting with 30% ethanol, and discarding eluate. Then eluting with 60-80% ethanol, and recovering solvent from the eluent to obtain the dry extract of the triterpenoid part of the thyme herb.
(3) Preparation of Dijiao phenolic part
1) Taking 100g of Japanese pepper, adding 40% ethanol according to the material-liquid ratio of 1:25 (g: mL), heating and refluxing for extraction for 40 minutes to obtain decoction and dregs.
2) Extracting the residue for 2 times according to the extraction operation in step 1). Mixing the extractive solutions obtained in step 1) and the above repeated extraction, filtering, concentrating, and drying to obtain dry extract.
3) Taking the dry extract obtained in the step 2), adding water to disperse the dry extract into a solution of 2 g/mL, and loading the solution onto an AB-8 macroporous resin column. Eluting with water, and removing water eluate. Eluting with 70% ethanol, and recovering solvent from the eluate to obtain dry extract of phenols part of herba Zanthoxyli.
(4) Grouping animals
50C 57/6N mice were bred in SPF environment, and after adaptive breeding for one week, the mice were divided into a normal group, a model group, a thyme herb total extract treatment group, a thyme herb triterpene part treatment group and a thyme herb phenol part treatment group, each of which was 10 mice.
(5) Animal modeling and administration
A liver injury model of a mouse is established by adopting a Lieber-Decalli alcohol liquid feed pairing feeding method. The normal group was fed with control liquid feed (no alcohol), the model group, the thyme herb total extract treatment group, the thyme herb triterpene site treatment group, and the thyme herb phenolic site treatment group were fed with alcohol liquid feed (4% V/V) for 6 weeks. During the period, the total extract of the Japanese pepper, the triterpenoid part of the Japanese pepper and the dry extract of the phenols part of the Japanese pepper are respectively given to each treatment group by 120 mg/kg every day.
(6) Detection method
After 6 weeks of experiment, the mice in each group are anesthetized by chloral hydrate, blood is taken and placed in a centrifuge tube, centrifugation is carried out at 4000 r/min for 15min, and serum is separated. ALT, AST, TC and TG levels in serum are detected by a Beckmann full-automatic biochemical analyzer.
3. Results
As shown in tables 2-3, compared with the normal group of mice, the levels of ALT, AST, TC and TG in the serum of the model group of mice are all obviously increased (P is less than 0.01), which indicates that the liver has abnormal functions and cell damage, and the liver damage model is successfully established. In the three groups of treatment groups, the serum biochemical index level of the thyme herb total extract treatment group is reduced most obviously compared with that of the model group, and the serum ALT, AST, TC and TG levels of mice in the thyme herb total extract treatment group are obviously lower than those of the thyme herb triterpene part treatment group and the thyme herb phenol part treatment group (P is less than 0.05 or P is less than 0.01).
TABLE 2 serum transaminase level test results for each group of mice
Group of ALT (U/L) AST (U/L)
Normal group 32.00±5.14 130.20±7.39
Model set 74.00±15.99## 181.12±18.43##
Pepper total extract treatment group 39.50±4.99** 142.14±7.72**
Thyme herb triterpene part treatment group 51.40±9.66△ 162.24±18.62△△
Dijiao phenolic part treatment group 56.00±9.58△△ 159.17±12.50△
Note: # P <0.05, # P <0.01 compared to normal; comparing P <0.05, P <0.01 to model groups; compared with the treatment group of the total extract of the thymus serpyllum, the delta P is less than 0.05, and the delta P is less than 0.01.
TABLE 3 serum TC and TG level assay results in various groups of mice
Group of TC (mmol/L) TG (mmol/L)
Normal group 3.48±0.38 2.17±0.21
Model set 4.08±0.21## 2.78±0.40##
Pepper total extract treatment group 3.58±0.32** 2.38±0.27*
Thyme herb triterpene part treatment group 3.88±0.22△ 2.80±0.45△
Dijiao phenolic part treatment group 3.79±0.22△ 2.85±0.42△
Note: comparison with Normal group # P<0.05，##P<0.01; comparison with model group<0.05，**P<0.01; compared with the treatment group of the total extract of Japanese pepper△P<0.05，△△P<0.01。
4. Conclusion
The research result shows that the anti-liver injury activity of the total extract of the thymus herb is obviously superior to that of the triterpene and the phenol of the thymus herb, and the liver protection activity of the thymus plant is proved to have synergistic effect.
The above is only a preferred embodiment of the present invention, and it should be noted that the above preferred embodiment should not be considered as limiting the present invention, and the protection scope of the present invention should be subject to the scope defined by the claims. It will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the spirit and scope of the invention, and these modifications and adaptations should be considered within the scope of the invention.

Claims (10)

1. Application of Thymus plant in preparing medicine for preventing and treating liver injury is provided.
2. Use of thymus plant according to claim 1 for the preparation of a medicament for the prevention and treatment of liver damage, characterized in that it comprises the following steps: extracting Thymus plant to obtain extract for preparing liver protecting medicine.
3. The thymus plant extract as claimed in claim 2, characterized by the raw materials: labiatae (Labi)atae) Thymus (Thymus)Thymus) The whole plant of the plant.
4. Thymus plant material according to claim 3, characterized in that preferred thymus plants comprise thyme, allogenic thyme, short-cut thyme, long-toothed thyme, Xingkai thyme, Xingan thyme, pseudo-thyme, line-leaf thyme, Altai thyme, oblique-leaf thyme, pulse-producing thyme, Black-Dragon-river thyme, short-hair thyme, mountain thyme, Rose thyme, Black-thyme, thyme variation of thyme, and Asian thyme.
5. A thymus plant extract characterized in that the preparation method of said extract comprises the following steps:
(1) adding ethanol into a thymus plant medicinal material, heating, refluxing and extracting, and filtering an extracting solution to obtain a first material;
(2) adding ethanol into the rest Thymus plant materials, repeatedly extracting for more than 0 times, and filtering the extractive solution to obtain a second material;
(3) and mixing the first material and the second material, recovering the solvent, and drying under reduced pressure to obtain the composite material.
6. The method for preparing an extract according to claim 5, wherein: the material-liquid ratio is thymus plant: ethanol 1:20 to 1:80 (g: mL); repeating the extraction for 1-3 times; the extraction time is 40-120 minutes each time; the concentration of the solvent ethanol used for extraction is 50-90%.
7. A method for producing a Thymus extract according to any one of claims 2 to 6.
8. Use of thymus plant and its extract as claimed in any one of claims 2 to 6 for the preparation of a medicament for the protection of the liver.
9. The application of thymus plants and thymus plant extracts as claimed in any one of claims 2 to 6 to the preparation of liver-protecting drugs, characterized in that: the dosage form of the medicine can be powder, granules, decoction, tablets, dispersible tablets, sustained-release tablets, sublingual tablets, orally disintegrating tablets, capsules, soft capsules, microcapsules, pills, dropping pills, concentrated pills, micro-pills, pellets, syrups, aerosols, injections, oral liquids, gels, microspheres, sustained-release preparations, controlled-release preparations and quick-release preparations.
10. The thymus extract as claimed in any one of claims 2 to 9, characterized in that it may further comprise pharmaceutically acceptable excipients.
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Citations (2)

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Publication number Priority date Publication date Assignee Title
CN106344648A (en) * 2016-09-26 2017-01-25 折改梅 Thymus plant extract and application thereof
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Publication number Priority date Publication date Assignee Title
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