WO2021179947A1 - 药物组合物在制备抗菌药物中的应用 - Google Patents

药物组合物在制备抗菌药物中的应用 Download PDF

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WO2021179947A1
WO2021179947A1 PCT/CN2021/078602 CN2021078602W WO2021179947A1 WO 2021179947 A1 WO2021179947 A1 WO 2021179947A1 CN 2021078602 W CN2021078602 W CN 2021078602W WO 2021179947 A1 WO2021179947 A1 WO 2021179947A1
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pharmaceutical composition
pinellia
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贾振华
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石家庄以岭药业股份有限公司
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Priority to ZA2022/08931A priority patent/ZA202208931B/en

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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
    • A61K36/17—Gnetophyta, e.g. Ephedraceae (Mormon-tea family)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • 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
    • 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)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/28—Asteraceae or Compositae (Aster or Sunflower family), e.g. chamomile, feverfew, yarrow or echinacea
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/34—Campanulaceae (Bellflower family)
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    • A61K36/18—Magnoliophyta (angiosperms)
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/70—Polygonaceae (Buckwheat family), e.g. spineflower or dock
    • A61K36/708—Rheum (rhubarb)
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/888—Araceae (Arum family), e.g. caladium, calla lily or skunk cabbage
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    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
    • A61K36/8966—Fritillaria, e.g. checker lily or mission bells
    • A—HUMAN NECESSITIES
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    • A—HUMAN NECESSITIES
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • 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
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
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    • 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
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    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
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    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates to the application of a pharmaceutical composition in the preparation of antibacterial drugs, and belongs to the field of medicine.
  • Antibiotics refer to a class of secondary metabolites produced by microorganisms (including bacteria, fungi, actinomycetes) or higher animals and plants during their lives that have anti-pathogen or other activities and can interfere with the development of other living cells. Chemical material. Commonly used clinical antibiotics include extracts from microbial culture fluids and chemically synthesized or semi-synthetic compounds. The antibacterial or bactericidal effect of antibiotics mainly includes four major mechanisms of action, namely: inhibition of bacterial cell wall synthesis, enhancement of bacterial cell membrane permeability, interference with bacterial protein synthesis, and inhibition of bacterial nucleic acid replication and transcription. Antibiotics play a major role in the treatment of many bacterial infectious diseases.
  • the present invention is an improved invention based on the pharmaceutical composition disclosed in the CN101549060B patent, and the contents recorded in the patent document are quoted here in full.
  • the aforementioned patent does not disclose that the pharmaceutical composition has an antibacterial effect.
  • the pharmaceutical composition of the present invention has excellent antibacterial effect
  • the pharmaceutical composition is made of the following raw materials in parts by weight:
  • Ephedra 52-86 Plaster 194-324 Forsythia 194-324 Scutellaria 78-130 Mulberry 194-324 Bitter almond 78-130 Qianhu 78-130 Pinellia 78-130 Chenpi 78-130 Fritillaria 78-130 Burdock seed 78 -130 Honeysuckle 78-130 Rhubarb 39-65 Platycodon 46-76 Licorice 39-65.
  • the weight ratio of the bulk drug of the pharmaceutical composition of the present invention is preferably:
  • the weight ratio of the bulk drug of the pharmaceutical composition of the present invention is also preferably:
  • the weight ratio of the bulk drug of the pharmaceutical composition of the present invention is also preferably:
  • Ephedra 69 Gypsum 259; Forsythia 259; Scutellaria baicalensis 104; Mulberry bark 259; Bitter almond 104; Qianhu 104; Pinellia 104; Licorice 52.
  • the weight ratio of the bulk drug of the pharmaceutical composition of the present invention is also preferably:
  • the bitter almond is fried bitter almond
  • the fritillary is Fritillaria vulgaris
  • the honeysuckle is Lonicera japonica
  • the pinellia is Qing Pinellia.
  • the pharmaceutical composition of the present invention has been confirmed by clinical experiments to effectively kill various bacteria, and the effect is remarkable.
  • the active ingredient of the pharmaceutical composition of the present invention can be made by a method comprising the following steps:
  • step D Mix the fine powder obtained in step A and the clear ointment mixture obtained in step C to obtain the active ingredient of the pharmaceutical composition.
  • the dosage form of the medicine of the present invention can be capsule, tablet, powder, granule, oral liquid, soft capsule, pill, tincture, syrup, suppository, gel, spray or injection.
  • compositions such as fillers, disintegrants, lubricants, suspending agents, binders, sweeteners, correctives, and preservatives. , Substrate, etc.
  • Fillers include: starch, pregelatinized starch, lactose, mannitol, chitin, microcrystalline cellulose, sucrose, etc.; disintegrants include: starch, pregelatinized starch, microcrystalline cellulose, sodium carboxymethyl starch, Cross-linked polyvinylpyrrolidone, low-substituted hydroxypropyl cellulose, croscarmellose sodium, etc.; lubricants include: magnesium stearate, sodium lauryl sulfate, talc, silicon dioxide, etc.; suspending agent Including: polyvinylpyrrolidone, microcrystalline cellulose, sucrose, agar, hydroxypropyl methylcellulose, etc.; binders include starch slurry, polyvinylpyrrolidone, hydroxypropyl methylcellulose, etc.; sweeteners include: Saccharin sodium, aspartame, sucrose, cyclamate, glycyrrhetinic acid, etc.; correctives include:
  • the tablet can be made by a method including the following steps:
  • step D Spray drying the cleansing paste mixture obtained in step C, and collect the spray powder for later use;
  • step E The spray powder obtained in step D and the fine powder obtained in step A are prepared by using ethanol as a binder to prepare a soft material, and then sieved to granulate; it is obtained by pressing according to the conventional method of pharmacy.
  • the preferred preparation method of the tablet may be a method including the following steps:
  • step D Spray drying the cleansing paste mixture obtained in step C, and collect the spray powder for later use;
  • step E The spray powder obtained in step D and the fine powder obtained in step A are used to prepare a soft material with ethanol as a binder, sieved, granulated, dried, and then sieved, and then mixed with sodium starch glycolate, microcrystalline cellulose, and magnesium stearate. Evenly, press the tablet and get it.
  • the raw materials can be weighed in proportion and prepared by conventional preparation methods, for example, according to the preparation described in Fan Biting's "Chinese Medicine Pharmacy” (Shanghai Science Press, December 1997, 1st edition) Process, made into a pharmacologically acceptable conventional dosage form.
  • the pharmaceutical composition of the present invention has a significant antibacterial effect on type A hemolytic streptococcus, type B hemolytic streptococcus, Staphylococcus aureus and pneumococcus.
  • the bacterium is a gram-positive bacterium; more preferably, the gram-positive bacterium is selected from Staphylococcus aureus, streptococcus A hemolytic, streptococcus B hemolytic, and pneumococcus.
  • the bacterium is a gram-negative bacterium; more preferably, the gram-negative bacterium is selected from Escherichia coli and Shigella.
  • Figure 1 shows the protective effect of the pharmaceutical composition of the present invention on mice infected with Staphylococcus aureus.
  • step D Spray drying the cleansing paste mixture obtained in step C, and collect the spray powder for later use;
  • step E The spray powder obtained in step D and the fine powder obtained in step A are prepared using 80% ethanol as a binder to prepare a soft material, sieved and granulated, dried at 60 degrees, and then granulated. Add sodium carboxymethyl starch, microcrystalline cellulose, and magnesium stearate, mix well, and prepare tablets according to the conventional preparation method to obtain.
  • step D Spray drying the cleansing paste mixture obtained in step C, and collect the spray powder for later use;
  • step E The spray powder obtained in step D and the fine powder obtained in step A are prepared using 80% ethanol as a binder to prepare a soft material, sieved and granulated, dried at 60 degrees, and then granulated. Add sodium carboxymethyl starch, microcrystalline cellulose, and magnesium stearate, mix well, and prepare tablets according to the conventional preparation method to obtain.
  • step D Spray drying the cleansing paste mixture obtained in step C, and collect the spray powder for later use;
  • step E The spray powder obtained in step D and the fine powder obtained in step A are prepared by using 80% ethanol as a binder to prepare a soft material, sieved and granulated, dried at 60 degrees, sizing, adding sodium carboxymethyl starch and microcrystalline cellulose , Magnesium stearate is mixed uniformly, and it is made into tablets according to the conventional preparation method.
  • the API formula is:
  • step D Spray drying the cleansing paste mixture obtained in step C, and collect the spray powder for later use;
  • step E The spray powder obtained in step D and the fine powder obtained in step A are prepared using 80% ethanol as a binder to prepare a soft material, sieved and granulated, dried at 60 degrees, and then granulated. Add sodium carboxymethyl starch, microcrystalline cellulose, and magnesium stearate, mix well, and prepare tablets according to the conventional preparation method to obtain.
  • the above medicinal materials can be obtained as capsules according to conventional methods.
  • Ephedra 68g Plaster 215g Forsythia 215g Radix Scutellariae 100g Mulberry Bark 220g Bitter Almond 90g Qian Hu 90g Pinellia 90g Tangerine Peel 90g Fritillaria 90g Burdock Seed 90g Honeysuckle 90g Rhubarb 50g Platycodon 50g Licorice 50g
  • the above medicinal materials are prepared into granules according to conventional methods.
  • the above medicinal materials can be obtained as injections according to conventional methods.
  • the above medicinal materials can be obtained as pills according to conventional methods.
  • the pharmaceutical composition prepared in Example 3 of the present invention that is, the granules (hereinafter referred to as the drug of the present invention) after granulation in step E of Example 3 and before tableting (hereinafter referred to as the drug of the present invention) were performed as follows:
  • the test drug is the pharmaceutical composition of the present invention (prepared in Example 3), the pharmaceutical composition is granules, brown-yellow powder, and each gram of granules is equivalent to 4.095 g crude drug.
  • the dosage is 22g of crude drug per person per day, orally, the human body weight is calculated as 60kg, which is equivalent to 0.367g pharmaceutical composition/kg body weight.
  • the positive control drug is Shuanghuanglian Oral Liquid, produced by Harbin Pharmaceutical Group Sanjing Pharmaceutical Co., Ltd., and the batch number is 07101243.
  • Staphylococcus aureus (bacterial number 26112), A hemolytic streptococcus (bacterial number 32209), beta hemolytic streptococcus (bacterial number 32210), pneumococcus (bacterial number 31001), large intestine Bacillus (Bacterial No. 44155), Staphylococcus Epidermidis (Bacterial No. 26487), Catacoccus (Bacterial No. 29103), Pseudomonas aeruginosa (Bacterial No. 10104), Shigella (Bacterial No. 51592), Typhoid Bacillus (Bacterial No. 50071), All were purchased from the National Institute for the Control of Pharmaceutical and Biological Products.
  • Culture medium broth medium and 2% agar broth medium, for Staphylococcus aureus, Escherichia coli, Staphylococcus epidermidis, Pseudomonas aeruginosa, Shigella, Typhoid bacillus passage and test; 10% serum broth
  • the medium and 10% serum agar broth medium are used for the passage and test of type A hemolytic streptococcus, type B hemolytic streptococcus, and pneumococcus.
  • Preparation of the pharmaceutical composition of the present invention Weigh 4.88 g of the pharmaceutical composition particles of the present invention and dissolve them in 40 ml of physiological saline, which is equivalent to 500 mg crude drug/ml. After autoclaving and storing in a refrigerator at 4°C for 5 days, centrifuged at 5000 rpm for 20 minutes, and the supernatant was taken for in vitro experiments.
  • Table 1 In vitro antibacterial effect of the pharmaceutical composition of the present invention (diameter of inhibition zone mm)
  • (1)Drug dilution Take 8 test tubes and number them in sequence. Use a pipette to pipette 4ml of serum-containing broth medium according to the aseptic procedure and put them into each test tube except the first tube; the first tube does not add culture medium Instead, add 8ml of the original solution of the pharmaceutical composition of the present invention (500mg/ml), take out 4ml from the first tube and put it in the second tube, shake it up repeatedly; in the same way, add 4ml from the second tube and add it to the third tube , Shake well; repeat the above steps until the serial multiples are diluted to the eighth tube.
  • the titers of the drug dilution are stock solution, 1:1, 1:2, 1:4, 1:8, 1:16, 1:32, 1:64, and the sequential concentration of the pharmaceutical composition of the present invention is 500 mg/ml, 250mg/ml, 125mg/ml, 62.5mg/ml, 31.3mg/ml, 15.6mg/ml, 7.8mg/ml, 3.9mg/ml; Shuanghuanglian oral liquid was diluted in the same way.
  • Bacterial solution dilution Dilute the culture solution of pneumococcus, A-hemolytic streptococcus, acetyl-hemolytic streptococcus, Escherichia coli and Staphylococcus aureus cultured for 18 hours into 1:100 dilution.
  • Test method Take each 40 test tubes and add 1.8ml of broth medium or serum-containing broth medium, and then add 0.2ml of the pharmaceutical composition of the present invention and Shuanghuanglian of different dilutions in each test tube.
  • the final concentrations of the pharmaceutical composition of the present invention are respectively 50mg/ml, 25mg/ml, 12.5mg/ml, 6.25mg/ml, 3.13mg/ml, 1.56mg/ml, 0.78mg/ml, 0.39mg/ml, Shuanghuanglian Concentration cannot be expressed in oral liquid because the concentration is not indicated.
  • Bacterial inoculation Take 20 ⁇ l of diluted bacterial solution of pneumococcus, streptococcus alpha, and streptococcus acetyl into the serum-containing medium, and take 10 ⁇ l of Escherichia coli and Staphylococcus aureus culture solution and add serum-free Medium. Observe the growth of bacteria after incubating at 37°C for 24 hours. The lowest drug concentration for aseptic growth was taken as the minimum inhibitory concentration (MIC) of the drug for the strain. Pipette 10 ⁇ l of each tube culture that has never seen bacterial growth and transfer it to sterile medium. After incubating at 37°C for 24 hours, observe whether there is bacterial growth. If there is no bacterial growth, it means that the concentration is the minimum bactericidal concentration (MBC) of the drug. . Results Table 2 and Table 3.
  • Table 2 The minimum inhibitory concentration of the drug combination of the present invention against standard strains in vitro (test tube method)
  • Table 3 The minimum bactericidal concentration of the drug combination of the present invention against standard strains in vitro (test tube method)
  • test tube method 20 strains of gram-positive bacteria isolated clinically (among which Staphylococcus aureus, streptococcus alpha, hemolytic beta Streptococcus and Pneumococcus (5 strains each), the inhibitory effect of the pharmaceutical composition of the present invention on clinical strains was studied.
  • the test method is the same as above. The results are shown in Tables 4 and 5.
  • Table 4 Antibacterial effect of the pharmaceutical composition of the present invention on clinical strains (test tube method)
  • Table 5 The bactericidal effect of the pharmaceutical composition of the present invention on clinical strains (test tube method)
  • the pharmaceutical composition of the present invention has a minimum inhibitory concentration of 12.5 mg/ml for both Staphylococcus aureus and Pneumococcus, which is minimally bactericidal for both The concentration is 25mg/ml; the minimum inhibitory concentration for A hemolytic streptococcus is 50mg/ml; the minimum inhibitory concentration for B hemolytic streptococcus is 12.5-25mg/ml, and the minimum bactericidal concentration is 50mg/ml .
  • the pharmaceutical composition of the present invention has effective antibacterial effects against common gram-positive bacteria (A and B hemolytic streptococcus, pneumococcus, and staphylococcus aureus) that are common in respiratory tract infections.
  • common gram-positive bacteria A and B hemolytic streptococcus, pneumococcus, and staphylococcus aureus
  • test drug is the pharmaceutical composition of the present invention, which is the same as the in vitro antibacterial test study (1).
  • the test drug is prepared with 0.5% CMC-Na to the required concentration before use.
  • Gastric membrane purchased from Beijing Yidelong Trading Company. Weigh 5g of gastricin, put it in a mortar, add a small amount of normal saline, add it with the grinding, and finally add to 100ml, and sterilize it under 10 pounds of pressure for 10 minutes.
  • Staphylococcus aureus (bacteria number 26112), purchased from the National Institute for the Control of Pharmaceutical and Biological Products.
  • UV-120-02 ultraviolet-visible spectrophotometer is a product of Shimadzu, Japan, a 400R high-speed refrigerated centrifuge, and a product of Heraeus, Germany.
  • Staphylococcus aureus bacteria liquid Take Staphylococcus aureus, seed it once with broth medium, and then use broth medium to take the above bacterial culture for 6 hours, 37°C incubator After 16 hours of incubation in medium, centrifuge at 3000rpm ⁇ 10min, take the precipitate, dilute it with sterile saline, and compare the color with an ultraviolet-visible spectrophotometer at a wavelength of 640nm, adjust the OD value of the bacterial solution to 0.200, and take out a part of the bacterial solution Count the number of bacteria. Centrifuge the bacterial solution at 3000rpm ⁇ 10min and remove the supernatant. Use sterilized 5% gastrin to restore the bacteria to the original volume. Inject 0.4ml into the intraperitoneal cavity of each mouse, and the amount of bacteria injected into each mouse is about 1.6 ⁇ 10 8 indivual.
  • mice 92 ICR mice were randomly divided into 5 groups, namely the control group (gave an equal volume of 0.5% CMC-Na); Shuanghuanglian group (gave 10ml/kg, equivalent to a human daily dose 10 times the amount); the pharmaceutical composition group of the present invention is administered intragastrically, and the doses are respectively 1.9g/kg (equivalent to 5 times the daily dosage of a person), 3.7g/kg (equivalent to 10 times the daily dosage of a person), 7.4g/kg (equivalent to 20 times the human consumption).
  • the five groups were administered 1 hour before the injection of bacteria, 0.4ml/10g body weight, twice a day for 3 days.
  • Table 7 The protective effect of the drug combination of the present invention against Staphylococcus aureus infection
  • mice in the control group injected with Staphylococcus aureus reached 94% within 7 days.
  • mice in the large and medium-dose groups of the pharmaceutical composition of the present invention were reduced to 53% and 58%, respectively, which were significantly different from the control group (p ⁇ 0.01); although there was no statistical difference in the low-dose group, However, the mortality rate of mice is still decreasing, showing a dose-effect relationship as a whole.
  • the results show that the pharmaceutical composition of the present invention has a significant protective effect on the injection of Staphylococcus aureus in mice, and can delay the death time of the mice. .

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Abstract

一种药物组合物在制备抗菌药物中的应用，所述药物组合物由麻黄、石膏、连翘、黄芩、桑白皮、炒苦杏仁、前胡、清半夏、陈皮、浙贝母、牛蒡子、山银花、大黄、桔梗、甘草制成，用于抑制革兰氏阳性细菌金黄色葡萄球菌、甲型溶血型链球菌、乙型溶血型链球菌和肺炎双球菌，以及革兰氏阴性细菌大肠杆菌和痢疾杆菌。

Description

药物组合物在制备抗菌药物中的应用
本申请要求于2020年3月7日提交到中国专利局的发明名称为“一种中药组合物在制备抗菌药物中的应用”的中国专利申请202010153831.8的优先权，其内容通过引用以整体并入本文。
技术领域
本发明涉及一种药物组合物在制备抗菌药物中的应用，属于医药领域。
背景技术
抗生素，是指由微生物(包括细菌、真菌、放线菌属)或高等动植物在生活过程中所产生的具有抗病原体或其他活性的一类次级代谢产物，能干扰其他生活细胞发育功能的化学物质。临床常用的抗生素有微生物培养液中的提取物以及用化学方法合成或半合成的化合物。抗生素的抑菌或杀菌作用，主要包含四大作用机理，即：抑制细菌细胞壁合成，增强细菌细胞膜通透性，干扰细菌蛋白质合成以及抑制细菌核酸复制转录。抗生素在许多细菌感染性疾病的治疗上发挥着重大作用。然而，由于抗生素的滥用，导致越来越多的病菌开始产生耐药性。由于抗生素的使用存在着滥用导致的疗程长、随意更换、盲目使用以及多种并用的不规范治疗,导致了许多菌株出现多重耐药和耐药广泛的问题。
因此，目前对于有效的抗菌药物，尤其是对于具有广谱抗菌性能、能有效改善耐药性的抗菌药物依然存在着迫切需求。
本发明是在CN101549060B专利公开的药物组合物的基础上进行的改进发明，在此全文引用该专利文件记载的内容。上述专利并未公开该药物组合物有抗菌作用。
发明内容
本发明人在研究中意外发现，本发明的药物组合物具有优异的抗菌作用，，所述药物组合物由下列重量份的原料药制成：
麻黄52-86 石膏194-324 连翘194-324 黄芩78-130 桑白皮194-324 苦杏仁78-130 前胡78-130 半夏78-130 陈皮78-130 贝母78-130 牛蒡子78-130 金银花78-130 大黄39-65 桔梗46-76 甘草39-65。
本发明药物组合物的原料药的重量份比优选为：
麻黄52；石膏324；连翘194；黄芩78；桑白皮194；苦杏仁130；前胡78；半夏130；陈皮78；贝母78；牛蒡子130；金银花130；大黄39；桔梗76；甘草65。
本发明药物组合物的原料药的重量份比还优选为：
麻黄86；石膏194；连翘324；黄芩130；桑白皮324；苦杏仁78；前胡130；半夏78；陈皮130；贝母130；牛蒡子78；金银花78；大黄65；桔梗46；甘草39。
本发明药物组合物的原料药的重量份比还优选为：
麻黄69；石膏259；连翘259；黄芩104；桑白皮259；苦杏仁104；前胡104；半夏104；陈皮104；贝母104；牛蒡子104；金银花104；大黄52；桔梗61；甘草52。
本发明药物组合物的原料药的重量份比还优选为：
麻黄55；石膏254；连翘318；黄芩107；桑白皮203；苦杏仁107；前胡82；半夏105；陈皮84；贝母125；牛蒡子122；金银花113；大黄42；桔梗60；甘草50。
优选地，在本发明的药物组合物中，所述苦杏仁为炒苦杏仁、贝母为浙贝母、金银花为山银花、半夏为清半夏。
本发明药物组合物经临床实验证实能有效杀灭多种细菌，效果显著。
本发明药物组合物的活性成分可以由包括以下步骤的方法制成:
A、按组方比例称取浙贝母，粉碎成细粉，备用；
B、按组方比例称取麻黄、连翘、炒苦杏仁、清半夏、牛蒡子、大黄加40-70％乙醇回流提取2次，每次1-4小时，第一次加8-10倍 量，第二次加6-9倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.14-1.16的清膏，备用；
C、按组方比例称取石膏、桑白皮、前胡、陈皮、山银花、桔梗、甘草，加水煎煮两次，每次1-4小时，第一次加9-11倍量，第二次加7-9倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.14-1.16的清膏，并与步骤B所得的清膏合并，得清膏混合物，备用；
D、将步骤A所得细粉和步骤C所得清膏混合物混合，得到该药物组合物的活性成分。
本发明药物的剂型可以为胶囊剂、片剂、散剂、颗粒剂、口服液、软胶囊、丸剂、酊剂、糖浆剂、栓剂、凝胶剂、喷雾剂或注射剂。
为使上述剂型能够实现，优选在制备这些剂型时加入药学可接受的辅料，例如：填充剂、崩解剂、润滑剂、助悬剂、粘合剂、甜味剂、矫味剂、防腐剂、基质等。填充剂包括：淀粉、预胶化淀粉、乳糖、甘露醇、甲壳素、微晶纤维素、蔗糖等；崩解剂包括：淀粉、预胶化淀粉、微晶纤维素、羧甲基淀粉钠、交联聚乙烯吡咯烷酮、低取代羟丙纤维素、交联羧甲基纤维素钠等；润滑剂包括：硬脂酸镁、十二烷基硫酸钠、滑石粉、二氧化硅等；助悬剂包括：聚乙烯吡咯烷酮、微晶纤维素、蔗糖、琼脂、羟丙基甲基纤维素等；粘合剂包括，淀粉浆、聚乙烯吡咯烷酮、羟丙基甲基纤维素等；甜味剂包括：糖精钠、阿斯帕坦、蔗糖、甜蜜素、甘草次酸等；矫味剂包括：甜味剂及各种香精；防腐剂包括：尼泊金类、苯甲酸、苯甲酸钠、山梨酸及其盐类、苯扎溴铵、醋酸氯乙定、桉叶油等；基质包括：PEG6000，PEG4000，虫蜡等。
其中片剂可以采用包括如下步骤的方法制成：
A、按组方比例称取贝母，粉碎成细粉，备用；
B、按组方比例称取麻黄、连翘、苦杏仁、半夏、牛蒡子、大黄，加40-70％乙醇回流提取2次，每次1-4小时，第一次加8-10倍量，第二次加6-9倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.14-1.16的清膏，备用；
C、按组方比例称取石膏、桑白皮、前胡、陈皮、金银花、桔梗、 甘草，加水煎煮两次，每次1-4小时，第一次加9-11倍量，第二次加7-9倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.14-1.16的清膏，并与步骤B所得的清膏合并，得清膏混合物，备用；
D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
E、将步骤D所得喷雾粉与步骤A所得细粉，以乙醇为黏合剂制软材，过筛制粒；按药学常规方法压片即得。
优选的片剂的制备方法可以是包括以下步骤的方法：
A、按组方比例称取贝母，粉碎成细粉，备用；
B、按组方比例称取麻黄、连翘、苦杏仁、半夏、牛蒡子、大黄，加50％乙醇回流提取2次，每次3小时，第一次加10倍量，第二次加6倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.15的清膏，备用；
C、按组方比例称取石膏、桑白皮、前胡、陈皮、金银花、桔梗、甘草，加水煎煮两次，每次2小时，第一次加10倍量，第二次加7倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.15的清膏，与步骤B所得的清膏合并，得清膏混合物，备用；
D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
E、将步骤D所得喷雾粉与步骤A所得细粉，以乙醇为黏合剂制软材，过筛制粒干燥后整粒，加入羧甲淀粉钠、微晶纤维素、硬脂酸镁，混匀，压片即得。
要制备得到本发明药物的其他剂型，可以按比例称取原料药，采用常规的制备方法制备，例如，按照范碧亭《中药药剂学》(上海科学出版社1997年12月第1版)记载的制备工艺，制成药剂学可接受的常规剂型。
实验证明，本发明的药物组合物对甲型溶血性链球菌、乙型溶血性链球菌、金黄色葡萄球菌和肺炎双球菌都有显著的抑菌作用。
在本发明的另一方面，还涉及抑制或杀灭细菌的方法，包括向有此需要的对象施用本发明的药物组合物。在本发明的另一方面，还涉及预防细菌感染或杀灭细菌的方法，包括向有此需要的对象施用本发 明的药物组合物。优选地，所述细菌为革兰氏阳性细菌；更优选地，所述革兰氏阳性细菌选自金黄色葡萄球菌、甲型溶血型链球菌、乙型溶血型链球菌和肺炎双球菌。在另一个优选的方面，所述细菌为革兰氏阴性细菌；更优选地，所述革兰氏阴性细菌选自大肠杆菌和痢疾杆菌。
附图说明
图1显示本发明的药物组合物对金黄色葡萄球菌感染的小鼠的保护作用。
具体实施方式
下述实施例用于举例说明本发明药物组合物的制备，但应理解其不对本发明的范围构成限制。
实施例1
处方：
麻黄52克；石膏324克；连翘194克；黄芩78克；桑白皮194克；苦杏仁130克；前胡78克；半夏130克；陈皮78克；浙贝母78克；牛蒡子130克；山银花130克；大黄39克；桔梗76克；甘草65克。
制备方法：
A、按组方比例称取浙贝母，粉碎成细粉，备用；
B、按组方比例称取麻黄、连翘、苦杏仁、半夏、牛蒡子、大黄，加50％乙醇回流提取2次，每次3小时，第一次加10倍量，第二次加6倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.15的清膏，备用；
C、按组方比例称取石膏、桑白皮、前胡、陈皮、山银花、桔梗、甘草，加水煎煮两次，每次2小时，第一次加10倍量，第二次加7倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.15的清膏，并与步骤B所得的清膏合并，得清膏混合物，备用；
D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
E、将步骤D所得喷雾粉与步骤A所得细粉，以80％乙醇为黏合剂制备软材，过筛制粒，60度干燥后，整粒。加入羧甲基淀粉钠、微晶纤维素、硬脂酸镁混匀，按常规制剂方法制成片剂，即得。
实施例2
处方：
麻黄86克；石膏194克；连翘324克；黄芩130克；桑白皮324克；炒苦杏仁78克；前胡130克；半夏78克；陈皮130克；贝母130克；牛蒡子78克；山银花78克；大黄65克；桔梗46克；甘草39克。
制备方法：
A、按组方比例称取贝母，粉碎成细粉，备用；
B、按组方比例称取麻黄、连翘、炒苦杏仁、半夏、牛蒡子、大黄，加40％乙醇回流提取2次，每次4小时，第一次加8倍量，第二次加9倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.14的清膏，备用；
C、按组方比例称取石膏、桑白皮、前胡、陈皮、山银花、桔梗、甘草，加水煎煮两次，每次4小时，第一次加9倍量，第二次加7倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.16的清膏，并与步骤B所得的清膏合并，得清膏混合物，备用；
D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
E、将步骤D所得喷雾粉与步骤A所得细粉，以80％乙醇为黏合剂制备软材，过筛制粒，60度干燥后，整粒。加入羧甲基淀粉钠、微晶纤维素、硬脂酸镁混匀，按常规制剂方法制成片剂，即得。
实施例3
处方：
麻黄69克；石膏259克；连翘259克；黄芩104克；桑白皮259克；炒苦杏仁104克；前胡104克；清半夏104克；陈皮104克；浙贝母104克；牛蒡子104克；山银花104克；大黄52克；桔梗61 克；甘草52克。
制备方法：
A、按组方比例称取浙贝母，粉碎成细粉，备用；
B、按组方比例称取麻黄、连翘、炒苦杏仁、清半夏、牛蒡子、大黄，加70％乙醇回流提取2次，每次1小时，第一次加10倍量，第二次加6倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.16的清膏，备用；
C、按组方比例称取石膏、桑白皮、前胡、陈皮、山银花、桔梗、甘草，加水煎煮两次，每次1小时，第一次加11倍量，第二次加7倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.14的清膏，并与步骤B所得的清膏合并，得清膏混合物，备用；
D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
E、将步骤D所得喷雾粉与步骤A所得细粉，以80％乙醇为黏合剂制备软材，过筛制粒，60度干燥后，整粒，加入羧甲基淀粉钠、微晶纤维素、硬脂酸镁混匀，按常规制剂方法制成片剂，即得。
实施例4：
原料药配方为：
麻黄55克；石膏254克；连翘318克；黄芩107克；桑白皮203克；炒苦杏仁107克；前胡82克；半夏105克；陈皮84克；浙贝母125克；牛蒡子122克；山银花113克；大黄42克；桔梗60克；甘草50克。
制备方法：
A、按组方比例称取浙贝母，粉碎成细粉，备用；
B、按组方比例称取麻黄、连翘、炒苦杏仁、半夏、牛蒡子、大黄，加60％乙醇回流提取2次，每次2小时，第一次加9倍量，第二次加7倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.15的清膏，备用；
C、按组方比例称取石膏、桑白皮、前胡、陈皮、山银花、桔梗、 甘草，加水煎煮两次，每次2.5小时，第一次加10倍量，第二次加7倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.14的清膏，并与步骤B所得的清膏合并，得清膏混合物，备用；
D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
E、将步骤D所得喷雾粉与步骤A所得细粉，以80％乙醇为黏合剂制备软材，过筛制粒，60度干燥后，整粒。加入羧甲基淀粉钠、微晶纤维素、硬脂酸镁混匀，按常规制剂方法制成片剂，即得。
实施例5：
麻黄62克；石膏220克；连翘256克；黄芩90克；桑白皮300克；苦杏仁90克；前胡90克；半夏90克；陈皮100克；贝母100克；牛蒡子100克；金银花100克；大黄50克；桔梗66克；甘草50克。
以上药材，按常规方法制成胶囊剂即得。
实施例6：
麻黄68克 石膏215克 连翘215克 黄芩100克 桑白皮220克 苦杏仁90克 前胡90克 半夏90克 陈皮90克 贝母90克 牛蒡子90克 金银花90克 大黄50克 桔梗50克 甘草50克
以上药材，按常规方法制成颗粒剂即得。
实施例7：
麻黄50克；石膏200克；连翘300克；黄芩100克；桑白皮250克；苦杏仁100克；前胡100克；半夏100克；陈皮100克；贝母100克；牛蒡子100克；金银花100克；大黄50克；桔梗50克；甘草50克。
以上药材，按常规方法制成注射剂即得。
实施例8：
麻黄60克；石膏200克；连翘200克；黄芩95克；桑白皮230克；苦杏仁95克；前胡95克；半夏95克；陈皮95克；贝 母95克；牛蒡子95克；金银花95克；大黄50克；桔梗50克；甘草50克。
以上药材，按常规方法制成丸剂即得。
生物活性实验例：
为证实本发明药物组合物具有抗菌效果，采用本发明实施例3制得的药物组合物(即实施例3步骤E整粒后、压片前的颗粒(以下称本发明药物)，进行了以下药理试验研究：
(一)本发明药物组合物体外抗菌试验研究
实验材料
1、药品与试剂：
(1)受试药为本发明药物组合物(实施例3制得)，所述药物组合物为颗粒，棕黄色粉末，每克颗粒相当于4.095克生药。用量为每日生药22g/人，口服，人体重以60kg计，折合0.367g药物组合物/kg体重。
(2)阳性对照药为双黄连口服液，哈药集团三精制药股份有限公司生产，批号为07101243。
2、细菌株：
(1)标准菌株：金黄色葡萄球菌(菌号26112)，甲型溶血性链球菌(菌号32209)，乙型溶血性链球菌(菌号32210)，肺炎双球菌(菌号31001)，大肠杆菌(菌号44155)，表皮葡萄球菌(菌号26487)，卡它球菌(菌号29103)，绿脓杆菌(菌号10104)，痢疾杆菌(菌号51592)，伤寒杆菌(菌号50071)，均购自中国药品生物制品检定所。
(2)临床菌株：金黄色葡萄球菌、甲型溶血性链球菌、乙型溶血性链球菌、肺炎双球菌各5株，以上20株细菌由北京中医药大学附属东方医院临床分离所得。
3、培养基：肉汤培养基和2％琼脂肉汤培养基，供金黄色葡萄球菌、大肠杆菌、表皮葡萄球菌、绿脓杆菌、痢疾杆菌、伤寒杆菌传代和试验用；10％血清肉汤培养基和10％血清琼脂肉汤培养基，供甲型 溶血性链球菌、乙型溶血性链球菌、肺炎双球菌传代和试验用。
方法与结果
1、本发明药物组合物的配制：称取4.88g的本发明药物组合物颗粒，溶于40ml生理盐水中，相当于500mg生药/ml。高压灭菌置4℃冰箱冷藏5日后，5000rpm离心20min，取上清供体外试验用。
2、本发明药物组合物对标准菌株的抑菌作用(钢环法)：抑菌试验用的各种标准菌株，分别用肉汤培养基和血清肉汤培养基传代二次，然后用肉汤培养基将各菌18小时培养物作10 -2稀释，取10 -2稀释菌液100μl，分别滴加于2％琼脂肉汤平皿和10％血清琼脂肉汤平皿，用灭菌L型玻璃棒在此平皿上均匀铺开，在平皿间隔一定部位放上5个灭菌不锈钢环(直径7mm)，在各钢环内分别滴加无菌生理盐水(阴性对照)及受试药液，即双黄连原液、本发明药物组合物原液(500mg/ml)、本发明药物组合物1:1稀释液(250mg/ml)、本发明药物组合物1:2稀释液(125mg/ml)，37℃培养箱中培养18小时后，测抑菌环直径(mm)。根据抑菌圈大小，判断药物的抑菌效果。结果如下表1所示。
表1：本发明药物组合物的体外抗菌作用(抑菌圈直径mm)
Figure PCTCN2021078602-appb-000001
Figure PCTCN2021078602-appb-000002
注：“—”表示无抑菌圈
2、本发明药物组合物体外对标准菌株的抑制作用(试管法)：以上试验的结果表明本发明药物组合物对革兰氏阳性菌株有一定的抑制作用，故本试验选用标准四株革兰氏阳性菌(金黄色葡萄球菌、甲性溶血型链球菌、乙性溶血型链球菌、肺炎双球菌)和一株革兰氏阴性菌(大肠杆菌)进行试验。
(1)药物稀释：取8支试管依次编号，按无菌操作程序用移液管吸取含血清的肉汤培养基4ml放入除第一管以外的各试管中；第一管不加培养基，而是加8ml的本发明药物组合物原液(500mg/ml)，从第一管中取出4ml放入第二管中，反复摇匀；同样，再从第二中吸取4ml加入第三管中，反复摇匀；重复以上步骤，直至系列倍比稀释到第八管。药物稀释的滴度依次为原液、1:1、1:2、1:4、1:8、1:16、1:32、1:64，本发明药物组合物的依次浓度为500mg/ml、250mg/ml、125mg/ml、62.5mg/ml、31.3mg/ml、15.6mg/ml、7.8mg/ml、3.9mg/ml；双黄连口服液作同样稀释。
(2)菌液稀释：将培养18h的肺炎双球菌、甲性溶血型链球菌、乙性溶血型链球菌、大肠杆菌和金黄色葡萄球菌培养液10μl+1ml肉汤作1:100稀释。
(3)试验方法：各取40只试管加肉汤培养基或含血清的肉汤培养基1.8ml，再在试管中加不同稀释度的本发明药物组合物和双黄连0.2ml，各试管中本发明药物组合物的终浓度分别为50mg/ml、25mg/ml、12.5mg/ml、6.25mg/ml、3.13mg/ml、1.56mg/ml、0.78mg/ml、0.39mg/ml，双黄连口服液因未标明浓度，故无法用浓度表示。细菌接种：取肺炎双球菌、甲性溶血型链球菌、乙性溶血型链球菌的稀释菌液20μl加入含血清的培养基中，取大肠杆菌和金黄色葡萄球菌培养液10μl加入不含血清的培养基中。37℃培养24h后观察细菌生长情况。以无菌生长的最低药物浓度作为药物对该菌株的最小抑菌浓度(MIC)。从未见细菌生长的各管培养物中分别吸取10μl转种于无菌培养基中，37℃培养24h后观察有无细菌生长，无细菌生长则表示该浓度为药物的最小杀菌浓度(MBC)。结果表2、表3。
表2：本发明药物组合物体外对标准菌株的最小抑菌浓度(试管法)
Figure PCTCN2021078602-appb-000003
表3：本发明药物组合物体外对标准菌株的最小杀菌浓度(试管法)
Figure PCTCN2021078602-appb-000004
3、本发明药物组合物体外对临床菌株的抑制作用(试管法)：试验选用20株临床分离所得的革兰氏阳性菌(其中金黄色葡萄球菌、甲性溶血型链球菌、乙性溶血型链球菌、肺炎双球菌各5株)，进行本发明药物组合物对临床菌株的抑制作用的研究。试验方法同上。结果见表4，5。
表4：本发明药物组合物对临床菌株的抑菌作用(试管法)
Figure PCTCN2021078602-appb-000005
表5：本发明药物组合物对临床菌株的杀菌作用(试管法)
Figure PCTCN2021078602-appb-000006
结论：
(1)采用钢环法研究本发明药物组合物体外抗标准菌株显示，本发明药物组合物体外对标准菌株中的金黄色葡萄球菌、甲性溶血型链球菌、乙性溶血型链球菌、肺炎双球菌、表皮葡萄球菌、卡他球菌、大肠杆菌、痢疾杆菌均有抑制作用。
(2)采用试管法研究本发明药物组合物体外抗标准菌株显示，本发明药物组合物对甲型溶血性链球菌的最小抑菌浓度和最小杀菌浓度分别为25mg/ml和50mg/ml，对金黄色葡萄球菌和乙型溶血性链球菌的最小抑菌浓度和最小杀菌浓度分别为12.5mg/ml和25mg/ml，对肺炎双球菌最小抑菌浓度和最小杀菌浓度分别为6.25mg/ml和12.5mg/ml。。
(3)采用试管法研究本发明药物组合物体外抗临床分离菌株显示，本发明药物组合物对金黄色葡萄球菌和肺炎双球菌最小抑菌浓度均为12.5mg/ml，对二者的最小杀菌浓度则为25mg/ml；对甲型溶血性链球菌的最小抑菌浓度为50mg/ml；对乙型溶血性链球菌的最小抑菌浓度为12.5～25mg/ml，最小杀菌浓度为50mg/ml。
由此可见，本发明药物组合物体外对呼吸道感染常见的革兰氏阳性细菌(甲、乙型溶血性链球菌，肺炎双球菌，金黄色葡萄球菌)均有有效的抗菌作用。
(二)本发明药物组合物体内抗菌试验研究
实验材料
1、动物：ICR小鼠，雌雄各半，体重18～22g，北京维通利华实验动物技术有限公司生产，许可证编号SCXK(京2005-2006)。
2、药品与试剂：
(1)受试药为本发明药物组合物，同体外抗菌试验研究(一)。所述受试药临用前用0.5％CMC-Na配成所需浓度使用。
(2)阳性对照药为双黄连口服液，同体外抗菌试验研究(一)。
(3)胃膜素，购于北京益得龙商贸公司。称取胃膜素5g，放入研钵内，加少量生理盐水，随磨随加，最后加至100ml，于10磅加压灭菌10分钟即可。
3、菌株：金黄色葡萄球菌(菌号26112)，购自中国药品生物制品检定所。
4、仪器：UV-120-02型紫外-可见分光光度计为日本岛津产品，400R型高速冷冻离心机，德国Heraeus产品。
方法与结果
1、金黄色葡萄球菌菌液的制备、注射及计数：取金黄色葡萄球菌，用肉汤培养基传种一次，再用肉汤培养基取培养6小时的上述细菌培养物，37℃培养箱中培养16小时后，离心3000rpm×10min，取沉淀，用灭菌生理盐水稀释，再用紫外-可见分光光度计在640nm波 长处比色，将细菌液的OD值调至0.200，取出部分菌液作菌数计数。细菌液离心3000rpm×10min后去上清，用灭菌后的5％胃膜素将细菌恢复至原体积，每只小鼠腹腔注射0.4ml，每只小鼠注射细菌量约为1.6×10 8个。
2、动物分组及处理：92只ICR小鼠随机分为5组，即对照组(灌胃给等体积0.5％CMC-Na)；双黄连组(灌胃给10ml/kg，相当于人日用量的10倍)；本发明药物组合物组，灌胃给药，剂量分别为1.9g/kg(相当于人日用量的5倍)、3.7g/kg(相当于人日用量的10倍)、7.4g/kg(相当于人用量的20倍)。此五组于注射细菌前1小时给药，0.4ml/10g体重，每日2次，持续3天。连续观察一周，第一天每3小时观察1次，之后每天观察2次。结果表明，本发明药物组合物大、中剂量对小鼠体内金黄色葡萄球菌致死量感染有显著的保护作用，并能推迟小鼠的死亡时间。统计学处理采用SPSS软件进行卡方检验。结果见表6、7和图1。
表6：本发明药物组合物体内抗金黄色葡萄球菌感染的作用
Figure PCTCN2021078602-appb-000007
表7：本发明药物组合物体内对金黄色葡萄球菌感染的保护作用
Figure PCTCN2021078602-appb-000008
Figure PCTCN2021078602-appb-000009
注：与对照组比较，*p<0.05，**p<0.01。
结果与结论：
(1)注射金黄色葡萄球菌的对照组小鼠7天内死亡率达94％。
(2)口服双黄连后小鼠死亡率降至50％，与对照组比较具有显著性差异(p<0.01)，表明双黄连对小鼠体内注射金黄色葡萄球菌具有显著的保护作用。
(3)本发明药物组合物大、中剂量组小鼠死亡率分别降至53％和58％，与对照组比较均有显著性差异(p<0.01)；小剂量组虽无统计学差异，但小鼠死亡率仍有降低趋势，总体上呈现量效关系。结果表明本发明药物组合物对小鼠体内注射金黄色葡萄球菌具有显著的保护作用，并能推迟小鼠死亡时间。。

Claims (14)

  1. 一种药物组合物在制备抗菌药物中的应用，其中所述药物组合物由下列重量份的原料药制成：
    麻黄52-86 石膏194-324 连翘194-324 黄芩78-130 桑白皮194-324 苦杏仁78-130 前胡78-130 半夏78-130 陈皮78-130 贝母78-130 牛蒡子78-130 金银花78-130 大黄39-65 桔梗46-76 甘草39-65。
  2. 根据权利要求1所述的应用，其中所述药物组合物由下列重量份的原料药制成：
    麻黄52 石膏324 连翘194 黄芩78 桑白皮194 苦杏仁130 前胡78 半夏130 陈皮78 贝母78 牛蒡子130 金银花130 大黄39 桔梗76 甘草65。
  3. 根据权利要求1所述的应用，其中所述药物组合物由下列重量份的原料药制成：
    麻黄86；石膏194；连翘324；黄芩130；桑白皮324；苦杏仁78；前胡130；半夏78；陈皮130；贝母130；牛蒡子78；金银花78；大黄65；桔梗46；甘草39。
  4. 根据权利要求1所述的应用，其中所述药物组合物由下列重量份的原料药制成：
    麻黄69；石膏259；连翘259；黄芩104；桑白皮259；苦杏仁104；前胡104；半夏104；陈皮104；贝母104；牛蒡子104；金银花104；大黄52；桔梗61；甘草52。
  5. 根据权利要求1所述的应用，其中所述药物组合物由下列重量份的原料药制成：
    麻黄55；石膏254；连翘318；黄芩107；桑白皮203；苦杏仁107；前胡82；半夏105；陈皮84；贝母125；牛蒡子122；金银花113；大黄42；桔梗60；甘草50。
  6. 根据权利要求1-5任一项所述的应用，其中所述苦杏仁为炒苦杏仁、贝母为浙贝母、金银花为山银花、半夏为清半夏。
  7. 根据权利要求1-6任一项所述的应用，其中所述药物组合物的活性成分由以下步骤制成：
    A、按组方比例称取贝母，粉碎成细粉，备用；
    B、按组方比例称取麻黄、连翘、苦杏仁、半夏、牛蒡子、大黄，加40-70％乙醇回流提取2次，每次1-4小时，第一次加8-10倍量，第二次加6-9倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.14-1.16的清膏，备用；
    C、按组方比例称取石膏、桑白皮、前胡、陈皮、金银花、桔梗、甘草，加水煎煮两次，每次1-4小时，第一次加9-11倍量，第二次加7-9倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.14-1.16的清膏，并与步骤B所得的清膏合并，得清膏混合物；
    D、将步骤A所得细粉和步骤C所得的清膏混合物混合，得到该药物组合物的活性成分。
  8. 根据权利要求1-6任一项所述的应用，其中所述药物剂型为片剂、胶囊剂、散剂、颗粒剂、口服液、丸剂、酊剂、糖浆剂、栓剂、凝胶剂、喷雾剂或注射剂的形式。
  9. 根据权利要求8所述的应用，其中所述片剂是由包括以下步骤的方法制成的：
    A、按组方比例称取贝母，粉碎成细粉，备用；
    B、按组方比例称取麻黄、连翘、苦杏仁、半夏、牛蒡子、大黄，加40-70％乙醇回流提取2次，每次1-4小时，第一次加8-10倍量，第二次加6-9倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.14-1.16的清膏，备用；
    C、按组方比例称取石膏、桑白皮、前胡、陈皮、金银花、桔梗、甘草，加水煎煮两次，每次1-4小时，第一次加9-11倍量，第二次加7-9倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.14-1.16的清膏，并与步骤B所得的清膏合并，得清膏混合物；
    D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
    E、将步骤D所得喷雾粉与步骤A所得细粉，以乙醇为黏合剂制软材， 过筛制粒；压片。
  10. 根据权利要求9所述的应用，其中所述片剂是由包括以下步骤的方法制成的：
    A、按组方比例称取贝母，粉碎成细粉，备用；
    B、按组方比例称取麻黄、连翘、苦杏仁、半夏、牛蒡子、大黄，加50％乙醇回流提取2次，每次3小时，第一次加10倍量，第二次加6倍量，提取液合并，滤过，滤液减压回收乙醇，浓缩至60℃热测相对密度为1.15的清膏，备用；
    C、按组方比例称取石膏、桑白皮、前胡、陈皮、金银花、桔梗、甘草，加水煎煮两次，每次2小时，第一次加10倍量，第二次加7倍量，煎液合并，滤过，浓缩至60℃热测相对密度为1.15的清膏，并与步骤B所得的清膏合并，得清膏混合物；
    D、将步骤C所得的清膏混合物喷雾干燥，收集喷雾粉备用；
    E、将步骤D所得喷雾粉与步骤A所得细粉，以乙醇为黏合剂制软材，过筛制粒干燥后整粒，加入羧甲淀粉钠、微晶纤维素、硬脂酸镁，混匀，压片。
  11. 根据权利要求1-5任一项所述的应用，其中该药物用于抑制革兰氏阳性细菌感染。
  12. 根据权利要求11所述的应用，其中所述革兰氏阳性细菌选自金黄色葡萄球菌、甲型溶血型链球菌、乙型溶血型链球菌和肺炎双球菌。
  13. 根据权利要求1-5任一项所述的应用，其中所述药物用于抑制革兰氏阴性细菌感染。
  14. 根据权利要求13所述的应用，其中所述革兰氏阴性细菌选自大肠杆菌和痢疾杆菌。
PCT/CN2021/078602 2020-03-07 2021-03-02 药物组合物在制备抗菌药物中的应用 WO2021179947A1 (zh)

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