CN113350440B - Application of traditional Chinese medicine composition in preparation of antibacterial drugs - Google Patents
Application of traditional Chinese medicine composition in preparation of antibacterial drugs Download PDFInfo
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- CN113350440B CN113350440B CN202010153831.8A CN202010153831A CN113350440B CN 113350440 B CN113350440 B CN 113350440B CN 202010153831 A CN202010153831 A CN 202010153831A CN 113350440 B CN113350440 B CN 113350440B
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Classifications
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- A—HUMAN NECESSITIES
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- 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
- A61K36/17—Gnetophyta, e.g. Ephedraceae (Mormon-tea family)
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- A—HUMAN NECESSITIES
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/06—Aluminium, calcium or magnesium; Compounds thereof, e.g. clay
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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
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Abstract
The invention discloses an application of a traditional Chinese medicine composition in preparation of antibacterial medicines. The traditional Chinese medicine composition mainly comprises ephedra, gypsum, fructus forsythiae, radix scutellariae, cortex mori radicis, bitter apricot kernel, peucedanum root and other medicinal herbs, plays the integral regulation advantage of the compound traditional Chinese medicine, has the characteristics of multiple directions, multiple layers and multiple targets, and has remarkable effect by in-vivo and in-vitro practical verification.
Description
Technical Field
The invention relates to an application of a traditional Chinese medicine composition in preparation of antibacterial medicines, and belongs to the field of Chinese herbal medicines.
Background
Antibiotics, which are chemical substances that interfere with the developmental functions of other living cells, are secondary metabolites with antipathogenic or other activities produced by microorganisms (including bacteria, fungi, actinomycetes) or higher animals and plants during life. Extracts from microbial cultures of antibiotics commonly used in clinic and chemically synthesized or semisynthetic compounds.
Antibacterial or bactericidal action of antibiotics and other antibacterial agents is mainly aimed at killing the mechanism of 'bacteria but no people (or other animals and plants'), and comprises four action mechanisms, namely: inhibiting bacterial cell wall synthesis, enhancing bacterial cell membrane permeability, interfering with bacterial protein synthesis, and inhibiting bacterial nucleic acid replication transcription.
When people have minor diseases such as cold, diarrhea, cough and the like at ordinary times, antibiotics are immediately applied to treat the diseases, and the diseases are quickly removed. However, abuse of antibiotics, which causes more and more pathogens to begin to develop resistance, is not a smart drug, and none inhibits or kills all bacteria. With the application of a large number of antibiotics and the continuous development of new products, bacterial resistance is also growing year by year. As a result of human abuse of antibiotics, the generation of the first generation of resistant bacteria takes only 2 years, and the development of a new antibiotic by medical workers takes about 10 years, the development speed of antibiotics has far not followed the propagation speed of resistant bacteria.
In recent years, antibiotics play a great role in the treatment of many bacterial infectious diseases, but have the problems of long treatment course, random replacement and blind use and various combined irregular treatments caused by abuse, so that multiple drug resistance and wide drug resistance of many strains occur. The traditional Chinese medicine with excellent antibacterial effect provides a new direction and thought for solving the problem of bacterial drug resistance.
The traditional Chinese medicine antibiotics are a large class of traditional Chinese medicines which can inhibit and kill part of bacterial viruses, can regulate immunity in multiple links and have the efficacy of improving drug resistance to different degrees. The traditional Chinese medicine prescription is usually compounded by monarch drugs, ministerial drugs, adjuvant drugs and conductant drugs, the antibacterial traditional Chinese medicine prescription is synergistic antibacterial among the drugs, and the traditional Chinese medicine compound preparation is also compound antibacterial of various traditional Chinese medicines. The Shuanghuanglian oral liquid is a Chinese patent medicine with broad-spectrum antibacterial effect, and has good killing effect on various bacteria clinically. At present, a plurality of traditional Chinese medicine antibacterial medicines exist, and the antibacterial effects of different compound antibacterial agents on the same strain are different, so that the optimal compound antibacterial agents need to be explored and found.
The traditional Chinese medicine composition has an antibacterial effect. The present invention is an improved invention based on the application number 2008100894475, which is incorporated by reference in its entirety. The above patent does not disclose that the Chinese medicinal composition has antibacterial effect.
Disclosure of Invention
The invention provides application of a traditional Chinese medicine composition in preparation of antibacterial medicines, wherein the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:
52-86 parts of ephedra, 194-324 parts of gypsum, 194-324 parts of weeping forsythia, 78-130 parts of baical skullcap root, 194-324 parts of white mulberry root-bark
78-130 parts of bitter apricot kernel, 78-130 parts of peucedanum root, 78-130 parts of pinellia tuber, 78-130 parts of dried orange peel, 78-130 parts of fritillary bulb and 78-130 parts of
78-130 parts of burdock, 78-130 parts of honeysuckle, 39-65 parts of rheum officinale, 46-76 parts of platycodon grandiflorum and 39-65 parts of liquorice.
The traditional Chinese medicine composition comprises the following raw materials in parts by weight:
herba Ephedrae 52; gypsum 324; fructus forsythiae 194; radix Scutellariae 78; cortex Mori 194; bitter apricot seed 130; radix Peucedani 78; pinellia ternate 130; dried orange peel 78; fritillary bulb 78; fructus Arctii 130; honeysuckle 130; rhubarb 39; radix Platycodi 76; licorice 65.
The traditional Chinese medicine composition also preferably comprises the following raw materials in parts by weight:
herba Ephedrae 86; gypsum 194; fructus forsythiae 324; radix Scutellariae 130; cortex Mori 324; bitter apricot kernel 78; radix Peucedani 130; pinellia tuber 78; dried orange peel 130; fritillary 130; burdock 78; honeysuckle 78; rhubarb 65; radix Platycodi 46; licorice 39.
The traditional Chinese medicine composition also preferably comprises the following raw materials in parts by weight:
ephedra 69; gypsum 259; fructus forsythiae 259; radix Scutellariae 104; cortex Mori 259; bitter almonds 104; radix Peucedani 104; pinellia ternate 104; dried orange peel 104; fritillary 104; fructus Arctii 104; honeysuckle 104; rhubarb 52; radix Platycodi 61; licorice 52.
The traditional Chinese medicine composition also preferably comprises the following raw materials in parts by weight:
herba Ephedrae 55; gypsum 254; fructus forsythiae 318; radix Scutellariae 107; white mulberry root bark 203; bitter apricot kernel 107; radix Peucedani 82; pinellia ternate 105; dried orange peel 84; fritillary bulb 125; burdock 122; honeysuckle 113; rhubarb 42; radix Platycodi 60; licorice root 50.
In the traditional Chinese medicine composition, the bitter apricot kernel is fried bitter apricot kernel, the fritillaria is thunberg fritillary bulb, the honeysuckle is lonicera japonica and the pinellia tuber is purified pinellia tuber.
The traditional Chinese medicine composition mainly comprises ephedra, gypsum, fructus forsythiae, radix scutellariae, cortex mori radicis and the like, plays the integral regulation advantage of the compound traditional Chinese medicine, has the characteristics of multiple directions, multiple layers and multiple targets, and has remarkable effect in clinical practice, and the actual effect can effectively kill various bacteria.
The traditional Chinese medicine can be replaced by traditional Chinese medicines with the same or similar effects, and the medicines can be processed according to national traditional Chinese medicine processing standards or Chinese medicine dictionary.
The active ingredients of the traditional Chinese medicine composition are prepared by the following steps:
A. weighing Bulbus Fritillariae Thunbergii according to the proportion of the prescription, and pulverizing into fine powder for use;
B. weighing herba Ephedrae, fructus forsythiae, parched semen Armeniacae amarum, purified pinellia Tuber, fructus Arctii, and radix et rhizoma Rhei according to the proportion, reflux extracting with 40-70% ethanol for 2 times, each for 1-4 hr, adding 8-10 times of the extractive solution for the first time and adding 6-9 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.14-1.16 at 60deg.C;
C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water and decocting for two times, 1-4 hours each time, adding 9-11 times of the weight of the first time and 7-9 times of the weight of the second time, merging decoction, filtering, concentrating to obtain clear paste with the relative density of 1.14-1.16 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;
the fine powder obtained in the step A and the combined fluid extract obtained in the step C jointly form the active ingredient of the pharmaceutical composition.
The medicament of the invention is in the dosage forms of capsules, tablets, powder, granules, oral liquid, soft capsules, pills, tinctures, syrups, suppositories, gels, sprays or injections.
To enable the above dosage forms, pharmaceutically acceptable excipients are added in the preparation of these dosage forms, for example: fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, flavoring agents, preservatives, matrices, and the like. The filler comprises: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, and the like; the disintegrating agent comprises: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, crosslinked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, crosslinked sodium carboxymethyl cellulose, and the like; the lubricant comprises: magnesium stearate, sodium lauryl sulfate, talc, silica, and the like; the suspending agent comprises: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, and the like; the binder includes starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.; the sweetener comprises: saccharin sodium, aspartame, sucrose, sodium cyclamate, glycyrrhetinic acid, etc.; the flavoring agent comprises: sweetener and various flavors; the preservative comprises: nipagin, benzoic acid, sodium benzoate, sorbic acid and salts thereof, benzalkonium bromide, chlorhexidine acetate, eucalyptus oil and the like; the matrix comprises: PEG6000, PEG4000, insect wax, and the like.
Wherein the tablet is prepared by the following steps:
A. weighing Bulbus Fritillariae Cirrhosae according to the proportion of the prescription, and pulverizing into fine powder for use;
B. weighing herba Ephedrae, fructus forsythiae, semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei according to the formula ratio, reflux extracting with 40-70% ethanol for 2 times, each for 1-4 hr, adding 8-10 times of the extractive solution for the first time and adding 6-9 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.14-1.16 at 60deg.C;
C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, honeysuckle, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water for decoction twice, each time for 1-4 hours, adding 9-11 times of the weight of the first time and 7-9 times of the weight of the second time, merging decoction, filtering, concentrating to obtain clear paste with the relative density of 1.14-1.16 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;
D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;
E. the spray powder obtained in the step D and the fine powder obtained in the step A are used for preparing soft materials by taking ethanol as an adhesive, and sieving and granulating are carried out; tabletting according to conventional pharmaceutical method.
The preparation method of the preferred tablet comprises the following steps:
A. weighing Bulbus Fritillariae Cirrhosae according to the proportion of the prescription, and pulverizing into fine powder for use;
B. weighing herba Ephedrae, fructus forsythiae, semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei according to the formula ratio, reflux extracting with 50% ethanol for 2 times, each for 3 hr, adding 10 times of the extractive solution for the first time, adding 6 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.15 at 60deg.C;
C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, honeysuckle, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water for decoction twice, each time for 2 hours, adding 10 times of the weight for the first time, adding 7 times of the weight for the second time, merging the decoction, filtering, concentrating to obtain clear paste with the relative density of 1.15 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;
D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;
E. and D, preparing soft materials by taking ethanol as an adhesive from the spray powder obtained in the step A, sieving, granulating, drying, finishing, adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, uniformly mixing, and tabletting. The preparation method of other dosage forms of the medicine comprises the following steps: the raw materials are weighed according to the proportion and prepared by adopting a conventional preparation method, for example, a preparation process recorded in Fan Biting traditional Chinese medicine pharmacy (1 st edition of 1997 of Shanghai science publishing Co., ltd.) is adopted to prepare a pharmaceutically acceptable conventional dosage form.
The traditional Chinese medicine composition has an antibacterial effect on the A-type hemolytic streptococcus; the traditional Chinese medicine composition can have an antibacterial effect on the beta-hemolytic streptococcus; the traditional Chinese medicine composition has an antibacterial effect on staphylococcus aureus. The Chinese medicinal composition has an antibacterial effect on pneumococci.
Description of the drawings:
fig. 1: the pharmaceutical composition has a protective effect on mice infected with staphylococcus aureus.
Detailed Description
The following examples are presented to illustrate the preparation of the medicaments of the present invention, but are not intended to limit the scope of the invention in any way.
Example 1
Prescription:
52 g of ephedra; 324 grams of gypsum; 194 g of weeping forsythia; 78 g of baikal skullcap root; 194 g of white mulberry root-bark; 130 g of bitter almond; radix peucedani 78 g; 130 g of pinellia ternate; 78 g of dried orange peel; 78 g of fritillary bulb; 130 g of burdock; 130 g of lonicera japonica; 39 g of rheum officinale; 76 g of platycodon grandiflorum; 65 g of liquorice.
The preparation method comprises the following steps:
A. weighing Bulbus Fritillariae Thunbergii according to the proportion of the prescription, and pulverizing into fine powder for use;
B. weighing herba Ephedrae, fructus forsythiae, semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei according to the formula ratio, reflux extracting with 50% ethanol for 2 times, each for 3 hr, adding 10 times of the extractive solution for the first time, adding 6 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.15 at 60deg.C;
C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water and decocting for 2 hours each time, adding 10 times of the weight for the first time and 7 times of the weight for the second time, merging the decoctions, filtering, concentrating to obtain clear paste with the relative density of 1.15 measured by heat at 60 ℃, merging with the clear paste obtained in the step B for standby;
D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;
E. and D, preparing the spray powder obtained in the step A, preparing soft materials by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, and finishing. Adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, mixing, and making into tablet by conventional method.
Example 2
Prescription:
86 g of ephedra; 194 g of gypsum; 324 g of fructus forsythiae; 130 g of radix scutellariae; 324 g of white mulberry root-bark; 78 g of fried bitter apricot kernel; 130 g of radix peucedani; 78 g of pinellia ternate; 130 g of dried orange peel; 130 g of fritillary bulb; 78 g of burdock; 78 g of lonicera japonica; 65 g of rheum officinale; 46 g of platycodon grandiflorum; licorice root, radix Glycyrrhizae Praeparata 39 g.
The preparation method comprises the following steps:
A. weighing Bulbus Fritillariae Cirrhosae according to the proportion of the prescription, and pulverizing into fine powder for use;
B. weighing herba Ephedrae, fructus forsythiae, parched semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei, reflux extracting with 40% ethanol for 2 times and 4 hr each time, adding 8 times of the extractive solution for the first time and 9 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.14 at 60deg.C;
C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water and decocting for two times, wherein the amount of the water is 9 times for each time, the amount of the water is 7 times for the first time, the decoction is combined, filtered and concentrated to the clear paste with the relative density of 1.16 measured by heat at 60 ℃ for standby;
D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;
E. and D, preparing the spray powder obtained in the step A, preparing soft materials by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, and finishing. Adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, mixing, and making into tablet by conventional method.
Example 3
Prescription:
69 g of ephedra; 259 grams of gypsum; 259 g of weeping forsythia; 104 g of radix scutellariae; 259 g of white mulberry root-bark; 104 g of fried bitter apricot kernel; 104 g of radix peucedani; 104 g of purified pinellia tuber; 104 g of dried orange peel; 104 g of fritillary bulb; 104 g of burdock; 104 g of lonicera japonica; 52 g of rheum officinale; 61 g of platycodon grandiflorum; licorice root, radix Glycyrrhizae Praeparata 52 g.
The preparation method comprises the following steps:
A. weighing Bulbus Fritillariae Thunbergii according to the proportion of the prescription, and pulverizing into fine powder for use;
B. weighing herba Ephedrae, fructus forsythiae, parched semen Armeniacae amarum, purified pinellia Tuber, fructus Arctii, and radix et rhizoma Rhei, reflux extracting with 70% ethanol for 2 times (1 hr each time), adding 10 times of the extractive solution for the first time, adding 6 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.16 at 60deg.C;
C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the prescription, adding water and decocting for two times, 1 hour each time, adding 11 times of the weight for the first time, adding 7 times of the weight for the second time, merging the decoctions, filtering, concentrating to obtain clear paste with the relative density of 1.14 measured by heat at 60 ℃, and merging with the clear paste obtained in the step B for standby;
D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;
E. and D, preparing the spray powder obtained in the step A, preparing the fine powder obtained in the step A by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, finishing, adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, uniformly mixing, and preparing into tablets according to a conventional preparation method.
Example 4:
the formula of the raw materials is as follows:
ephedra herb 55 g; 254 g of gypsum; 318 g of fructus forsythiae; baical skullcap root 107 g; 203 g of white mulberry root-bark; stir-frying 107 g of bitter apricot seeds; radix peucedani 82 g; 105 g of pinellia ternate; 84 g of dried orange peel; 125 g of fritillary bulb; 122 g of burdock; 113 g of lonicera japonica; 42 g of rheum officinale; 60 g of platycodon grandiflorum; 50 g of liquorice.
The preparation method comprises the following steps:
A. weighing Bulbus Fritillariae Thunbergii according to the proportion of the prescription, and pulverizing into fine powder for use;
B. weighing herba Ephedrae, fructus forsythiae, parched semen Armeniacae amarum, rhizoma Pinelliae, fructus Arctii, and radix et rhizoma Rhei, reflux extracting with 60% ethanol for 2 times and 2 hr each time, adding 9 times of the extractive solution for the first time and 7 times of the extractive solution for the second time, mixing the extractive solutions, filtering, recovering ethanol from the filtrate under reduced pressure, concentrating to obtain fluid extract with relative density of 1.15 at 60deg.C;
C. weighing gypsum, white mulberry root-bark, peucedanum root, dried orange peel, lonicera japonica, platycodon grandiflorum and liquorice according to the proportion of the formula, adding water and decocting for 2.5 hours each time, adding 10 times of the weight of the mixture for the first time, adding 7 times of the weight of the mixture for the second time, merging the decoctions, filtering, concentrating to obtain clear paste with the relative density of 1.14 measured by heat at 60 ℃, merging the clear paste obtained in the step B for later use;
D. c, spraying and drying the combined fluid extract obtained in the step C, and collecting spraying powder for later use;
E. and D, preparing the spray powder obtained in the step A, preparing soft materials by taking 80% ethanol as an adhesive, sieving, granulating, drying at 60 ℃, and finishing. Adding sodium carboxymethyl starch, microcrystalline cellulose and magnesium stearate, mixing, and making into tablet by conventional method.
Example 5:
62 g of ephedra, 220 g of gypsum, 256 g of weeping forsythia, 90 g of baikal skullcap root, 300 g of white mulberry root-bark
90 g of bitter apricot kernel, 90 g of peucedanum root, 90 g of pinellia tuber, 100 g of dried orange peel, 100 g of fritillary bulb, 100 g of burdock, 100 g of honeysuckle, 50 g of rhubarb, 66 g of platycodon root and 50 g of liquorice
The above materials are made into capsule by conventional method.
Example 6:
ephedra 68 g gypsum 215 g weeping forsythia 215 g baikal skullcap root 100 g mulberry bark 220 g
90 g of bitter apricot kernel, 90 g of peucedanum root, 90 g of pinellia tuber, 90 g of dried orange peel, 90 g of fritillary bulb and 90 g of
90 g of burdock, 90 g of honeysuckle, 50 g of rhubarb, 50 g of platycodon root, 50 g of liquorice
The above materials are made into granule by conventional method.
Example 7:
50 g of ephedra, 200 g of gypsum, 300 g of weeping forsythia, 100 g of baikal skullcap root, 250 g of mulberry bark
100 g of bitter apricot kernel, 100 g of peucedanum root, 100 g of pinellia tuber, 100 g of dried orange peel, 100 g of fritillary bulb and 100 g of
100 g of burdock, 100 g of honeysuckle, 50 g of rheum officinale, 50 g of platycodon grandiflorum, 50 g of liquorice
The above materials are made into injection by conventional method.
Example 8:
ephedra herb 60 g gypsum 200 g weeping forsythia fruit 200 g baikal skullcap root 95 g mulberry bark 230 g
Semen armeniacae amarae 95 g radix peucedani 95 g pinellia ternate 95 g dried orange peel 95 g fritillary bulb 95 g
95 g of burdock, 95 g of honeysuckle, 50 g of rhubarb, 50 g of platycodon root, 50 g of liquorice
The above materials are made into pill by conventional method.
Experimental example:
in order to confirm the antibacterial effect of the Chinese medicinal composition, the following pharmacological test study was carried out by the preparation method of the medicinal composition example 3, the granules before tabletting (hereinafter referred to as the medicinal composition of the invention), and the university of Beijing Chinese medicine was entrusted with:
in vitro antibacterial test study of the pharmaceutical composition of the invention
Experimental materials
1. Medicine and reagent:
(1) Test agent the pharmaceutical composition of the invention is provided by Shijia with the lot number 070601 by the company of Kaolin pharmaceutical Co., ltd. The test is carried out by using the medicinal composition particles, the property of the medicinal composition particles is brown yellow powder, and each gram of the particles is equivalent to 4.095 grams of crude drugs. The dosage is 22 g/person of crude drug per day, the tablet is taken orally, and the weight of human body is 60kg, which is equivalent to 0.367g/kg.
(2) The positive medicine is SHUANGHUANGLIAN oral liquid, manufactured by Sanjing medicine Co., ltd., and batch number is 07101243. The product is brown clear liquid, and has sweet and slightly bitter taste. Function and main indications: pungent and cool, has the effects of relieving exterior syndrome, clearing away heat and toxic materials. Is suitable for upper respiratory tract infection, pharyngitis, tonsillitis, etc. caused by bacterial infection. The drug amount per milliliter is not indicated in the specification, so the bacteriostasis test is expressed as the dilution multiple of the double coptis chinensis.
2. Bacterial strain:
(1) Standard strain: staphylococcus aureus (bacterium number 26112), streptococcus a haemolyticus (bacterium number 32209), streptococcus b haemolyticus (bacterium number 32210), pneumococcus (bacterium number 31001), escherichia coli (bacterium number 44155), staphylococcus epidermidis (bacterium number 26487), rhodococcus catarrhalis (bacterium number 29103), pseudomonas aeruginosa (bacterium number 10104), bacillus dysenteriae (bacterium number 51592), typhoid bacillus (bacterium number 50071) all purchased from the chinese medicine biologicals institute.
(2) Clinical strain: 5 strains of staphylococcus aureus, streptococcus A haemolyticus, streptococcus B haemolyticus and pneumococcus, more than 20 strains of bacteria are clinically separated from an eastern hospital affiliated to Beijing university of traditional Chinese medicine.
3. Culture medium: broth and 2% agar broth pour dishes for staphylococcus aureus, escherichia coli, staphylococcus epidermidis, pseudomonas aeruginosa, bacillus dysenteriae, typhoid bacillus passage and testing; 10% serum broth and 10% serum agar broth plates were used for passage and testing of Streptococcus hemolyticus A, streptococcus B, streptococcus pneumoniae.
Method and results
1. The preparation of the pharmaceutical composition comprises the following steps: 4.88g of the pharmaceutical composition particles of the present invention were weighed and dissolved in 40ml of physiological saline, which corresponds to 500mg crude drug/ml. High pressure sterilizing, refrigerating in 4 deg.c refrigerator for 5 days, centrifuging at 5000rpm for 20min, and collecting supernatant.
2. Antibacterial action of the pharmaceutical composition of the invention on standard strains (steel loop method): each standard strain used for bacteriostasis test was passaged twice with broth medium and serum broth medium, respectively, and then 18-hour culture of each strain was used as 10 with broth medium -2 Diluting to obtain 10 -2 100 mu L of diluted bacterial liquid is respectively dripped into a 2% agar broth plate and a 10% serum agar broth plate, the plates are evenly spread by using a sterilized L-shaped glass rod, 5 sterilized stainless steel rings (with the diameter of 7 mm) are placed at certain intervals on the plates, sterile physiological saline (negative control) and tested liquid medicine are respectively dripped into each steel ring, namely a Shuanghuanglian stock solution, a pharmaceutical composition stock solution of the invention (500 mg/ml), a pharmaceutical composition 1:1 diluent of the invention (250 mg/ml) and a pharmaceutical composition of the inventionThe clear pharmaceutical composition is diluted with 1:4 liquid (125 mg/ml), and after being cultured in an incubator at 37 ℃ for 18 hours, the diameter (mm) of a bacteriostasis ring is measured, and the bacteriostasis effect of the medicine is judged according to the size of the bacteriostasis ring. The results are shown in Table 1 below.
2. Inhibition of standard strains in vitro by the pharmaceutical composition of the invention (test tube method): according to the results of the above experiments, the pharmaceutical composition has a certain inhibition effect on gram-positive strains, so that four standard gram-positive bacteria (staphylococcus aureus, streptococcus alpha haemolyticus, streptococcus ethiosus and pneumococcus) and one gram-negative bacteria (escherichia coli) are selected for the experiment.
(1) Drug dilution: taking 8 test tubes, sequentially numbering, sucking 4ml of broth culture medium containing serum by a pipette according to a sterile operation procedure, putting the broth culture medium into each test tube, adding 8ml of stock solution (500 mg/ml) of the pharmaceutical composition of the invention into a first tube without adding the culture medium, taking out 4ml of the stock solution from the first tube, putting the stock solution into a second tube, and repeatedly shaking uniformly; also, 4ml of the mixture is sucked from the second part and added into a third pipe, and the mixture is repeatedly shaken uniformly; the above steps were repeated until serial fold ratios were diluted to the eighth tube. The titer of the medicine which is diluted in sequence is stock solution, 1:1, 1:2, 1:4, 1:8, 1:16, 1:32 and 1:64, and the concentration of the medicine composition of the invention is 500mg/ml, 250mg/ml, 125mg/ml, 62.5mg/ml, 31.3mg/ml, 15.6mg/ml, 7.8mg/ml and 3.9mg/ml in sequence; the Shuanghuanglian oral liquid is diluted as well.
(2) And (3) bacterial liquid dilution: culture broth of pneumococci, alpha-hemolytic streptococcus, beta-hemolytic streptococcus, E.coli and Staphylococcus aureus for 18h 10. Mu.l+1 ml broth was diluted 1:100.
(3) The test method comprises the following steps: 40 test tubes are added with 1.8ml of broth culture medium or broth culture medium containing serum, and then the test tubes are added with the pharmaceutical composition of the invention and 0.2ml of Shuanghuanglian with different dilutions, wherein the final concentration of the pharmaceutical composition of the invention in each test tube is respectively 50mg/ml, 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.13mg/ml, 1.56mg/ml, 0.78mg/ml and 0.39mg/ml, and the Shuanghuanglian oral liquid cannot be represented by the concentration because the concentration is not marked. Bacterial inoculation: 20. Mu.l of diluted bacteria liquid of pneumococcus, alpha hemolytic streptococcus and beta hemolytic streptococcus is added into a culture medium containing serum, and 10. Mu.l of culture liquid of escherichia coli and staphylococcus aureus is added into a culture medium without serum. After incubation at 37℃for 24h, bacterial growth was observed. The Minimum Inhibitory Concentration (MIC) of the drug against the strain was taken as the minimum drug concentration for sterile growth. Mu.l of each tube culture from which bacterial growth was not seen was aspirated and transferred to sterile medium, and after incubation at 37℃for 24h, bacterial growth was observed, and the absence of bacterial growth indicated that the concentration was the Minimum Bactericidal Concentration (MBC) of the drug. The results are shown in tables 2 and 3.
3. Inhibition of clinical strains in vitro by the pharmaceutical composition of the invention (test tube method): the test selects 20 gram positive bacteria (5 strains of staphylococcus aureus, streptococcus alpha haemolyticus, streptococcus beta haemolyticus and pneumococcus) obtained by clinical separation, and researches the inhibition effect of the pharmaceutical composition to clinical strains. The test method is the same as above. The results are shown in tables 4 and 5.
Knot (S)
(1) The in vitro anti-standard strain of the pharmaceutical composition is researched by adopting a steel loop method, and the pharmaceutical composition has an inhibition effect on staphylococcus aureus, alpha hemolytic streptococcus, beta hemolytic streptococcus, pneumococcus, staphylococcus epidermidis, catarrhalis, escherichia coli and shigella in the standard strain in vitro; has no inhibiting effect on Pseudomonas aeruginosa and typhoid bacillus.
(2) The test tube method researches on the in vitro anti-standard strain of the pharmaceutical composition show that the minimum antibacterial concentration and the minimum antibacterial concentration of the pharmaceutical composition are respectively 25mg/ml and 50mg/ml for the A-type hemolytic streptococcus, 12.5mg/ml and 25mg/ml for the staphylococcus aureus and the B-type hemolytic streptococcus, and 6.25mg/ml and 12.5mg/ml for the pneumococcus. The maximum concentration of the pharmaceutical composition of the present invention and Shuanghuanglian used in the test failed to inhibit the growth of E.coli.
(3) The test tube method researches on in vitro anti-clinical isolated strains of the pharmaceutical composition show that the minimum antibacterial concentration of the pharmaceutical composition is 12.5mg/ml for staphylococcus aureus and pneumococcus and 25mg/ml for both; the minimum antibacterial concentration of the streptococcus A is 50mg/ml, but the maximum concentration can not kill the streptococcus A; the minimum antibacterial concentration of the beta-hemolytic streptococcus is 12.5-25 mg/ml, and the minimum antibacterial concentration of the beta-hemolytic streptococcus is 50mg/ml.
In conclusion, the pharmaceutical composition has a certain antibacterial effect on gram-positive bacteria (streptococcus A and B hemolyticus, pneumococcus and staphylococcus aureus) common to respiratory tract infection in vitro.
(II) in vivo antibacterial test study of the pharmaceutical composition of the invention
Experimental materials
1. Animals: ICR mice, male and female halves, have a weight of 18-22 g, and are produced by Beijing vitamin Toril Hua laboratory animal technologies Co., ltd., license number SCXK (Beijing 2005-2006).
2. Medicine and reagent:
(1) Test agent the pharmaceutical composition of the invention is the same as in experiment one. Before the product is used, 0.5% CMC-Na is used for preparing the required concentration.
(2) The positive medicine is Shuanghuanglian oral liquid, and the same experiment is carried out.
(3) Gastric membranogen was purchased from the commercial company of Yideliong, beijing. Weighing 5g of gastric membranogen, placing into a mortar, adding a small amount of physiological saline, adding with grinding, adding 100ml, and sterilizing under 10 lbs under pressure for 10 min.
3. Strains: staphylococcus aureus (bacterial number 26112), purchased from the chinese pharmaceutical biologicals institute.
4. Instrument: UV-120-02 type ultraviolet-visible spectrophotometry was performed on the product of Shimadzu, japan, 400R high-speed refrigerated centrifuge, heraeus, germany.
Method and results
1. Preparation, injection and counting of staphylococcus aureus bacterial liquid: the staphylococcus aureus is inoculated once by a broth culture medium, the bacterial culture is cultivated for 6 hours by the broth culture medium, after the bacterial culture is cultivated for 16 hours in a 37 ℃ incubator, the bacterial culture is centrifugated for 3000rpm multiplied by 10 minutes, the precipitate is taken, diluted by sterilized normal saline, and is subjected to color comparison at 640nm by an ultraviolet-visible spectrophotometer, the OD value of bacterial liquid is regulated to 0.200, and part of bacterial liquid is taken out for counting bacterial count. Centrifuging the bacterial solution at 3000rpm×10min, removing supernatant, and recovering bacteria to original volume with sterilized 5% gastric membranogen, wherein each mouse is injected intraperitoneally with 0.4ml, and the bacterial injection amount of each mouse is about 1.6X10 8 And each.
2. Grouping and treating animals: 92 ICR mice were randomly divided into 5 groups, i.e., control group (gavage to equal volume of 0.5% CMC-Na); double coptis group (10 ml/kg for gastric lavage, equivalent to 10 times daily amount of people); the dosage of the pharmaceutical composition group of the invention is 1.9g/kg (corresponding to 5 times of daily dosage), 3.7g/kg (corresponding to 10 times of daily dosage) and 7.4g/kg (corresponding to 20 times of daily dosage). The five groups were dosed 1 hour prior to bacterial injection, 0.4ml/10g body weight, 2 times daily for 3 days. One week of continuous observation, 1 time every 3 hours on the first day, followed by 2 times per day. The results show that the large and medium doses of the pharmaceutical composition have remarkable protection effect on the lethal dose infection of staphylococcus aureus in mice, and can delay the death time of the mice. Statistical processing the Chi-Square Tests were performed using SPSS software. The results are shown in tables 6 and 7, FIG. 1.
Knot (S)
(1) The mortality rate of the staphylococcus aureus control group mice injected with the staphylococcus aureus control group reaches 94 percent within 7 days.
(2) The mortality of mice after oral administration of the double coptis is reduced to 50%, and compared with a control group, the double coptis has a remarkable difference (p < 0.01), which shows that the double coptis has a remarkable protective effect on staphylococcus aureus injected into the mice.
(3) The mortality of mice in the large and medium dose groups of the pharmaceutical composition is respectively reduced to 53 percent and 58 percent, compared with the control group, the pharmaceutical composition has significant difference (p < 0.01), and the mortality of mice in the small dose group also has a certain reduction trend, but no statistical difference. The result shows that the pharmaceutical composition has remarkable protective effect on the staphylococcus aureus injected into the mice in vivo, and can delay the death time of the mice. The pharmaceutical composition of the invention shows a certain dose-effect relationship in each dose group.
Claims (7)
1. The application of the traditional Chinese medicine composition in preparing antibacterial medicines is characterized in that the traditional Chinese medicine composition is prepared from the following raw materials in parts by weight:
52-86 parts of ephedra, 194-324 parts of gypsum, 194-324 parts of weeping forsythia, 78-130 parts of baical skullcap root, 194-324 parts of white mulberry root-bark
The application of the Chinese medicinal composition in preparing medicaments for resisting escherichia coli, staphylococcus epidermidis, catarrhalis and shigella dysenteriae is that the Chinese medicinal composition is applied to preparing medicaments for resisting escherichia coli, staphylococcus epidermidis, catarrhalis and shigella dysenteriae.
2. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:
ephedra 52 gypsum 324 weeping forsythia 194 baical skullcap root 78 white mulberry root-bark 194 bitter apricot seed 130 peucedanum root 78 pinellia tuber 130 tangerine peel 78 fritillary bulb 78 burdock 130 honeysuckle 130 rhubarb 39 platycodon root 76 liquoric root 65.
3. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:
herba Ephedrae 86; gypsum 194; fructus forsythiae 324; radix Scutellariae 130; cortex Mori 324; bitter apricot kernel 78; radix Peucedani 130; pinellia tuber 78; dried orange peel 130; fritillary 130; burdock 78; honeysuckle 78; rhubarb 65; radix Platycodi 46; licorice 39.
4. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:
ephedra 69; gypsum 259; fructus forsythiae 259; radix Scutellariae 104; cortex Mori 259; bitter almonds 104; radix Peucedani 104; pinellia ternate 104; dried orange peel 104; fritillary 104; fructus Arctii 104; honeysuckle 104; rhubarb 52; radix Platycodi 61; licorice 52.
5. The use according to claim 1, characterized by being prepared from the following raw materials in parts by weight:
herba Ephedrae 55; gypsum 254; fructus forsythiae 318; radix Scutellariae 107; white mulberry root bark 203; bitter apricot kernel 107; radix Peucedani 82; pinellia ternate 105; dried orange peel 84; fritillary bulb 125; burdock 122; honeysuckle 113; rhubarb 42; radix Platycodi 60; licorice root 50.
6. The use according to any one of claims 1-5, characterized in that the bitter almonds are fried bitter almonds, fritillary bulbs, honeysuckle flowers, lonicera confusa and pinellia ternate.
7. The use according to any one of claims 1-5, characterized in that the pharmaceutical dosage form is a tablet, capsule, powder, granule, oral liquid, pill, tincture, syrup, suppository, gel, spray.
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| PCT/CN2021/078602 WO2021179947A1 (en) | 2020-03-07 | 2021-03-02 | Use of pharmaceutical composition in preparing antibacterial drug |
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| 国家药典委员会编.中华人民共和国药典.中国医药科技出版社,2015,一部(2015),735. * |
| 连花急支片与抗生素治疗急性气管-支气管炎的随机对照临床研究;杨立波等;《中国医药导报》;第12卷(第19期);131页左栏第1段 * |
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