WO2021179616A1 - 小柴胡颗粒联合磷酸氯喹的新应用 - Google Patents

小柴胡颗粒联合磷酸氯喹的新应用 Download PDF

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WO2021179616A1
WO2021179616A1 PCT/CN2020/122715 CN2020122715W WO2021179616A1 WO 2021179616 A1 WO2021179616 A1 WO 2021179616A1 CN 2020122715 W CN2020122715 W CN 2020122715W WO 2021179616 A1 WO2021179616 A1 WO 2021179616A1
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traditional chinese
chinese medicine
medicine composition
ginger
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刘宏
张俊华
毕聪
郑如文
江志强
何逸禧
杜海泳
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广州白云山光华制药股份有限公司
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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)
    • A61K36/23—Apiaceae or Umbelliferae (Carrot family), e.g. dill, chervil, coriander or cumin
    • A61K36/233—Bupleurum
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A61K31/4706—4-Aminoquinolines; 8-Aminoquinolines, e.g. chloroquine, primaquine
    • 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)
    • A61K36/34—Campanulaceae (Bellflower family)
    • A61K36/344—Codonopsis
    • 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)
    • A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/484—Glycyrrhiza (licorice)
    • 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)
    • A61K36/53—Lamiaceae or Labiatae (Mint family), e.g. thyme, rosemary or lavender
    • A61K36/539—Scutellaria (skullcap)
    • 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)
    • A61K36/72—Rhamnaceae (Buckthorn family), e.g. buckthorn, chewstick or umbrella-tree
    • A61K36/725—Ziziphus, e.g. jujube
    • 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)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/888—Araceae (Arum family), e.g. caladium, calla lily or skunk cabbage
    • A61K36/8888—Pinellia
    • 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)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/906—Zingiberaceae (Ginger family)
    • A61K36/9068—Zingiber, e.g. garden ginger
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • 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
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants

Definitions

  • the invention relates to a new application of Xiao Chaihu granules combined with chloroquine phosphate, and belongs to the field of combined Chinese and Western medicine.
  • Mycoplasma pneumoniae is the pathogen of human primary atypical pneumonia and one of the most common pathogens of community-acquired pneumonia in our country.
  • Mycoplasma pneumoniae infection is mainly transmitted by droplets, which can cause upper and lower respiratory tract diseases such as pharyngitis, bronchitis, and acute pneumonia. It can also cause complications such as encephalitis, radiculitis, myocarditis, and hemolytic anemia.
  • Macrolide antibiotics are the first choice for its treatment. However, studies have shown that the resistance rate of Mycoplasma pneumoniae to macrolide antibiotics in vitro is significantly higher than that of other countries. Therefore, we should be vigilant and try other drug combinations.
  • lopinavir and ritonavir used to treat new coronavirus pneumonia are not superior to the control group in terms of improving clinical symptoms and accelerating virus clearance, and the effect is not significant . This may be because pneumonia is a complex disease with multiple system disorders. It is not effective to treat the disease only by actively attacking the virus.
  • the combination of Chinese and Western medicine is combined with anti-inflammatory, anti-viral or free radical scavenging when fighting infections. Defensive treatment may achieve better results.
  • Xiaochaihu Decoction is one of the eight famous prescriptions of traditional Chinese medicine.
  • Xiaochaihu Granules is a granular Chinese patent medicine preparation first developed by Guangzhou Baiyunshan Guanghua Pharmaceutical based on Xiaochaihu Decoction. It is formulated by the "Monarchs and Ministers", which has the effects of relieving the surface and heat, soothing the liver and stomach, and is used for exogenous diseases and evil offenses of Shaoyang syndrome.
  • Xiao Chaihu prescription has been proven to have many effects such as antipyretic, anti-inflammatory, regulating immunity, anti-liver fibrosis, anti-allergic and so on.
  • Xiao Chaihu granules have been effective in treating typhoid fever such as colds and flu.
  • Xiao Chaihu granules was listed as a special drug for the fight against SARS in Guangdong Province;
  • Xiao Chaihu granules were used to fight global "Influenza A"; in 2013, Xiao Chaihu granules were included in the diagnosis and treatment of dengue fever by the Guangdong Provincial Health Commission Guidelines for treatment and medication.
  • Chloroquine phosphate tablets are used to treat falciparum malaria, vivax malaria and vivax malaria that are sensitive to chloroquine.
  • chloroquine phosphate also has immunomodulatory activity, and can play antiviral and reduce lung injury effects through mechanisms such as inhibiting viral gene expression, changing the pH value of the endosome, and influencing autophagy.
  • Currently, cell experiments show that chloroquine phosphate can effectively inhibit the new coronavirus infection. It has been included in the "New Coronavirus Pneumonia Diagnosis and Treatment Plan (Trial Sixth Edition)" jointly issued by the National Health Commission and the State Administration of Traditional Chinese Medicine for use in the new Clinical treatment of pneumonia caused by coronavirus infection.
  • One of the objectives of the present invention is to provide a new application of a traditional Chinese medicine composition (Xiao Chaihu granules).
  • the active ingredients of the traditional Chinese medicine composition consist of 150 ⁇ 5 parts of Bupleurum spp., 56 ⁇ 5 parts of Scutellaria baicalensis Georgi, Ginger Pinellia 56 ⁇ 3 parts, Codonopsis 56 ⁇ 3 parts, Ginger 56 ⁇ 3 parts, Licorice 56 ⁇ 3 parts, Jujube 56 ⁇ 3 parts.
  • Another object of the present invention is to provide a new application of the above-mentioned traditional Chinese medicine composition combined with chloroquine phosphate.
  • the application of a traditional Chinese medicine composition combined with chloroquine phosphate in the preparation of a medicine for removing excess free radicals caused by Mycoplasma pneumoniae infection is composed of 150 ⁇ 5 parts of Bupleurum and 56 ⁇ 5 parts of Scutellaria baicalensis Georgi. 5 parts, ginger pinellia 56 ⁇ 3 parts, dangshen 56 ⁇ 3 parts, ginger 56 ⁇ 3 parts, licorice 56 ⁇ 3 parts, jujube 56 ⁇ 3 parts.
  • Another object of the present invention is to provide a combined medicine for removing excess free radicals caused by Mycoplasma pneumoniae infection.
  • a combined medicine for removing excessive free radicals caused by Mycoplasma pneumoniae infection comprising a traditional Chinese medicine composition and chloroquine phosphate, wherein the traditional Chinese medicine composition and the chloroquine phosphate become independent dosing units, or the traditional Chinese medicine composition and The chloroquine phosphate together form a combined dosing unit;
  • the weight parts of the active ingredients of the traditional Chinese medicine composition are composed of: Bupleurum vulgare 150 ⁇ 5 parts, Scutellaria baicalensis Georgi 56 ⁇ 5 parts, Jiang Pinellia 56 ⁇ 3 parts, and Codonopsis pilosulae 56 ⁇ 3 parts. , 56 ⁇ 3 parts of ginger, 56 ⁇ 3 parts of licorice, 56 ⁇ 3 parts of jujube.
  • Another object of the present invention is to also provide the application of the combined drug in preparing a drug for preventing and treating Mycoplasma pneumoniae.
  • the active ingredients of the traditional Chinese medicine composition are composed of 150 ⁇ 5 parts of Bupleurum spp., 56 ⁇ 5 parts of Scutellaria baicalensis Georgi, 56 ⁇ 3 parts of Scutellaria baicalensis Georgi. Parts, 56 ⁇ 3 parts of Codonopsis, 56 ⁇ 3 parts of ginger, 56 ⁇ 3 parts of licorice, 56 ⁇ 3 parts of jujube.
  • a combined medicine for the treatment of Mycoplasma pneumoniae comprising a traditional Chinese medicine composition and chloroquine phosphate, wherein the traditional Chinese medicine composition and the chloroquine phosphate become independent dosing units, or the traditional Chinese medicine composition and the chloroquine phosphate together form a combination
  • the active ingredient of the traditional Chinese medicine composition consists of 150 ⁇ 5 parts of Bupleurum, 56 ⁇ 5 parts of Scutellaria baicalensis, 56 ⁇ 3 parts of Pinellia ginger, 56 ⁇ 3 parts of Codonopsis, 56 ⁇ 3 parts of Ginger, 56 ⁇ 3 parts of licorice and 56 ⁇ 3 parts of jujube.
  • the active ingredients of the traditional Chinese medicine composition consist of 150 parts by weight of Bupleurum, 56 parts of Scutellaria baicalensis, 56 parts of Pinellia ginger, 56 parts of Codonopsis, 56 parts of ginger, 56 parts of licorice, and 56 parts of jujube. .
  • the traditional Chinese medicine composition is Xiao Chaihu granules.
  • the chloroquine phosphate is a tablet.
  • the present invention uses an SD rat model of Mycoplasma pneumoniae infection to carry out experiments, and it is found that the traditional Chinese medicine composition (Bupleurum, Scutellaria, Pinellia ginger, Codonopsis, ginger, licorice, jujube) can eliminate excessive free radicals caused by Mycoplasma pneumoniae infection.
  • the traditional Chinese medicine composition Bupleurum, Scutellaria, Pinellia ginger, Codonopsis, ginger, licorice, jujube
  • the combination of the traditional Chinese medicine composition (such as Xiao Chaihu granules) and chloroquine phosphate can significantly increase the serum levels of glutathione peroxidase, superoxide dismutase, and catalase, reduce the content of malondialdehyde, and Its effect is stronger than that of Xiao Chaihu granules and chloroquine phosphate alone, indicating that the combined administration can improve the body's ability to scavenge free radicals and reduce the cytotoxicity caused by free radical accumulation. It is found that Xiao Chaihu granules combined with chloroquine phosphate tablets can eliminate Mycoplasma pneumoniae infection.
  • Xiao Chaihu granules have the effect of removing Mycoplasma pneumoniae and sensitizing azithromycin.
  • Xiao Chaihu granules combined with chloroquine phosphate tablets have a better sensitizing effect on azithromycin.
  • Xiaochaihu granules combined with chloroquine phosphate can be used to prevent and treat Mycoplasma pneumoniae.
  • Figure 1 is a bar graph of the experimental results of each experimental group on the content of serum ROS in Example 2 of the present invention
  • Figure 2 is a bar graph showing the experimental results of each experimental group on the content of GSH-PX in serum in Example 2 of the present invention
  • Fig. 3 is a bar graph of the experimental results of the SOD content in serum of each experimental group in Example 2 of the present invention.
  • Figure 4 is a bar graph of the experimental results of each experimental group on the content of CAT in serum in Example 2 of the present invention
  • Figure 5 is a bar graph of the experimental results of each experimental group in Example 2 of the present invention on the content of MDA in the serum.
  • the "plurality” mentioned in the present invention means two or more.
  • “And/or” describes the association relationship of the associated objects, indicating that there can be three types of relationships, for example, A and/or B, which can mean: A alone exists, A and B exist at the same time, and B exists alone.
  • the character “/” generally indicates that the associated objects before and after are in an "or” relationship.
  • an application of a traditional Chinese medicine composition in the preparation of a medicine for removing excess free radicals caused by Mycoplasma pneumoniae infection.
  • the active ingredients of the traditional Chinese medicine composition are Bupleurum, Scutellaria, Pinellia ginger, Codonopsis and Ginger , Licorice, jujube.
  • the active ingredients of the traditional Chinese medicine composition consist of 150 parts by weight of Bupleurum vulgare, 56 parts of scutellaria baicalensis, 56 parts of ginger pinellia, 56 parts of Codonopsis, 56 parts of ginger, 56 parts of licorice, and jujube 56 copies.
  • the traditional Chinese medicine composition may be an ointment, a decoction or a concentrated pill, a mixture (including an oral liquid), a syrup, or a granule, etc. In a preferred embodiment, it is Granules.
  • the present invention relates to a combined drug for removing excess free radicals caused by Mycoplasma pneumoniae infection, comprising the above-mentioned traditional Chinese medicine composition and chloroquine phosphate, and the traditional Chinese medicine composition and the chloroquine phosphate are separately administered Unit, or the traditional Chinese medicine composition and the chloroquine phosphate together form a combined administration unit. That is, the traditional Chinese medicine composition (for example, small Bupleurum granules) and chloroquine phosphate can be administered separately or in combination.
  • the traditional Chinese medicine composition for example, small Bupleurum granules
  • chloroquine phosphate can be administered separately or in combination.
  • Chloroquine phosphate can be in the form of tablets or capsules, or other oral dosage forms. In a preferred embodiment, it is a tablet.
  • Xiao Chaihu granules have the effect of removing Mycoplasma pneumoniae and sensitizing azithromycin, and the combination of Xiao Chaihu granules and chloroquine phosphate tablets has better sensitizing effect on azithromycin.
  • Xiao Chaihu granules can be used to prepare medicines for the treatment of Mycoplasma pneumoniae, especially for assisting the application of azithromycin in the treatment of Mycoplasma pneumoniae.
  • Xiao Chaihu particles combined with chloroquine phosphate are used in the preparation of drugs for Mycoplasma pneumoniae.
  • Xiao Chaihu particles combined with chloroquine phosphate are used in the preparation of drugs for Mycoplasma pneumoniae assisting azithromycin.
  • the small Bupleurum granules in the following examples are sold by Guangzhou Baiyunshan Guanghua Pharmaceutical, and the specific active ingredients are: Bupleurum 150g, Scutellaria baicalensis Georgi 56g, Ginger Pinellia 56g, Codonopsis 56g, Ginger 56g, Licorice 56g, Jujube 56g, and the formulation is granules. .
  • the present invention found that the minimum inhibitory concentration of Xiao Chaihu granules against the standard strain of Mycoplasma pneumoniae FH was 3.125 mg/mL, and the minimum inhibitory concentration of chloroquine phosphate tablets against the standard strain of Mycoplasma pneumoniae FH was 8 mg/mL.
  • small Chaihu granules combined with azithromycin can significantly reduce the minimum inhibitory concentration of azithromycin (0.002mg/ML ⁇ 0.0005mg/mL), and small Chaihu granules + chloroquine phosphate tablets combined with azithromycin can significantly reduce the minimum inhibitory concentration of azithromycin (0.002mg/ml).
  • ML ⁇ 0.000125mg/mL indicating that Xiao Chaihu granules, Xiao Chaihu granules combined with azithromycin, Xiao Chaihu granules + chloroquine phosphate tablets combined with azithromycin can significantly inhibit Mycoplasma pneumoniae in vitro, that is, it is found that Xiao Chaihu granules can eliminate Mycoplasma pneumoniae and have the effect on azithromycin. Sensitization, Xiao Chaihu granules + chloroquine phosphate tablets have sensitization to azithromycin.
  • the standard strain MPFH of Mycoplasma pneumoniae was inoculated into the PPLO complete culture medium, and placed in a biochemical incubator containing CO 2 and saturated nitrogen at 37° C. When the culture medium turned from red to yellow, the subculture began.
  • the concentration of MPFH was quantitatively determined by CCU, and a bacterial solution with a concentration of 1 ⁇ 10 5 CCU/mL was taken for use.
  • Small Bupleurum Granules brew with boiling water to make them completely dissolve, sterilize and refrigerate to make 1000mg/mL small Bupleurum granules medicinal solution.
  • Chloroquine phosphate tablets add appropriate amount of water, ultrasonically shake, fully dissolve, sterilize and refrigerate to make 1280mg/mL chloroquine phosphate tablets liquid.
  • Azithromycin Prepare azithromycin solution with a final concentration of 1280 ⁇ g/mL under aseptic conditions.
  • MIC is an indicator to test whether a drug has the ability to inhibit the growth and reproduction of microorganisms in vitro.
  • MIC was determined on 96-well plates by liquid microdilution method. Use PPLO medium (containing phenol red indicator) to dilute each drug solution into 10 equal concentration gradients, and inoculate the same amount of MPFH bacterial solution (1 ⁇ 10 5 CCU/mL) so that the total volume of each well is 200 ⁇ L, each This sample was repeated three times. The hole without drug solution was used as a positive control, and the hole with medium added alone was used as a negative control. Drop in sterile liquid paraffin to seal, cover with cover, and incubate in 37°C incubator for 2-3 days.
  • the minimum drug solution concentration that does not grow Mycoplasma pneumoniae is the MIC value of the drug to Mycoplasma pneumoniae.
  • Xiao Chaihu granule combined with azithromycin administration group was added with minimal inhibitory concentration of Xiao Chaihu granules, set 10 concentrations of azithromycin, and record the MIC value of azithromycin after combined administration;
  • chloroquine phosphate combined with azithromycin administration group was added with minimal inhibitory concentration of phosphoric acid Chloroquine tablets, set 10 concentrations of azithromycin, record the MIC value of azithromycin after the combined administration;
  • Xiao Chaihu granules + chloroquine phosphate tablets combined with azithromycin administration group add the smallest inhibitory concentration of Xiao Chaihu particles, chloroquine phosphate tablets, set 10 concentrations Of azithromycin, record the
  • the minimum inhibitory concentrations of Xiao Chaihu granules, chloroquine phosphate tablets, azithromycin, Xiao Chaihu granules combined with azithromycin, chloroquine phosphate tablets combined with azithromycin, Xiao Chaihu granules + chloroquine phosphate tablets combined with azithromycin on MPFH are shown in Table 1.
  • Table 1 The results showed that the minimum inhibitory concentration of Xiao Chaihu granules against the standard strain of Mycoplasma pneumoniae FH was 3.125 mg/mL, and the minimum inhibitory concentration of chloroquine phosphate tablets against the standard strain of Mycoplasma pneumoniae FH was 8 mg/mL.
  • Xiao Chaihu granules combined with azithromycin can significantly reduce The minimum inhibitory concentration of azithromycin (0.002mg/ML ⁇ 0.0005mg/mL), Xiaochaihu granules+chloroquine phosphate tablets combined with azithromycin can significantly reduce the minimum inhibitory concentration of azithromycin (0.002mg/ML ⁇ 0.000125mg/mL).
  • the present invention uses an SD rat model of Mycoplasma pneumoniae infection to carry out experiments and finds that Xiao Chaihu granules, especially Xiao Chaihu granules combined with chloroquine phosphate tablets can significantly increase serum glutathione peroxidase, superoxide dismutase, and peroxide Hydrogenase level, reduce the content of malondialdehyde, and its effect is stronger than that of Xiao Chaihu granules and chloroquine phosphate alone, indicating that the combined administration can improve the body's ability to scavenge free radicals and reduce the cytotoxicity caused by free radical accumulation. That is, Xiao Chaihu granules are found
  • the combined chloroquine phosphate tablet has the purpose of eliminating excessive free radicals caused by Mycoplasma pneumoniae infection.
  • the standard strain MPFH of Mycoplasma pneumoniae was inoculated into the PPLO complete culture medium, and placed in a biochemical incubator containing CO 2 and saturated nitrogen at 37° C. When the culture medium turned from red to yellow, the subculture began.
  • the concentration of MPFH was quantitatively determined by CCU, and a bacterial solution with a concentration of 1 ⁇ 10 8 CCU/mL was taken for use.
  • SD rats were randomly divided into 10 groups, blank control group, Mycoplasma pneumoniae infection model group, positive drug azithromycin group (10mg/kg), chloroquine phosphate group (45mg/kg, the human clinical equivalent dose), low dose of Xiaochaihu granules Group (5.4g/kg, which is the human clinical equivalent dose), Xiao Chaihu granule medium dose group (10.8 g/kg), Xiao Chaihu granule high dose group (21.6 g/kg), Xiao Chaihu granules and chloroquine phosphate tablets combined administration Medicine low-dose group (Xiao Chaihu granules 5.4g/kg, chloroquine phosphate tablets 45mg/kg), combined administration middle-dose group (10.8g/kg, 45mg/kg), combined administration high-dose group (21.6g/kg, 45mg/kg), 10 animals per group.
  • the rats were lightly anesthetized with ether. Except for the blank control group, rats in each group were inoculated intranasally with 1 ⁇ 10 8 CCU/mL of MPFH bacterial solution, 100 ⁇ L per day for 4 days, and the blank control group was intranasally instilled with the same amount of normal saline .
  • the rats in each group began to intervene on the second day of the last vaccination.
  • Azithromycin was administered by subcutaneous injection, the Xiaochaihu granule group and the combined administration group were administered by gavage, and the blank control group and the model group were gavage with the same amount of normal saline.
  • ROS reactive oxygen species
  • GSH-PX glutathione peroxidase
  • SOD superoxide dismutase
  • CAT catalase
  • MDA malondialdehyde
  • ROS refers to the general term of oxygen-containing and active substances in the body or in the natural environment, including superoxide anion free radicals, hydroxyl free radicals, hydroperoxide free radicals, hydrogen peroxide, and lipid peroxides. ROS can oxidize protein, DNA and lipids, induce cell apoptosis, and directly participate in the pathological process of a variety of lung diseases, leading to lung damage and affecting alveolar repair.
  • GSH-PX can scavenge peroxides and hydroxyl free radicals in cells, block the chain reaction of lipid peroxidation, and protect the structure of cells. It is a kind of free radical scavenging enzyme widely present in the body.
  • SOD is the main antioxidant enzyme in the body, which can catalyze the disproportionation of superoxide anion radicals to generate oxygen and hydrogen peroxide, protecting cells from damage.
  • CAT is an enzyme that catalyzes the decomposition of hydrogen peroxide into oxygen and water, and is a free radical scavenger of hydrogen peroxide.
  • the MDA content can indirectly reflect the severity of the body's cells being attacked by free radicals.
  • the experimental results are shown in Figure 1 to Figure 5.
  • the experimental groups corresponding to each number are: 1- blank control group; 2- Mycoplasma pneumoniae infection model group; 3- positive drug azithromycin group; 4-chloroquine phosphate group; 5-Xiao Chaihu granules Low-dose group; 6-Xiao Chaihu granule medium-dose group; 7-Xiao Chaihu particle high-dose group; 8-co-administration low-dose group; 9-co-administration medium-dose group; 10-co-administration high-dose group.
  • the combined administration of Xiao Chaihu granules, chloroquine phosphate tablets, Xiao Chaihu granules and chloroquine phosphate tablets can significantly increase the levels of antioxidant enzymes GSH-PX, SOD and CAT and reduce ROS. , MDA content (P ⁇ 0.05, P ⁇ 0.01).
  • MDA content (P ⁇ 0.05, P ⁇ 0.01).
  • the levels of GSH-PX and SOD in the low, medium, and high-dose groups were significantly increased, and the ROS content was significantly reduced (P ⁇ 0.05, P ⁇ 0.01).
  • the MDA content was significantly different from that of the chloroquine phosphate group (P ⁇ 0.05, P ⁇ 0.01).
  • the GSH-PX level of the low, medium and high dose groups of the combined administration group was significantly higher than that of the Xiao Chaihu granule corresponding dose group, and the ROS content was significantly lower than that of the Xiao Chaihu granule corresponding dose group; the combined administration group
  • the levels of SOD and CAT in the middle and high dose groups were significantly different from the corresponding dose groups of Xiao Chaihu granules;
  • the MDA content of the combined dose group in the middle dose group was significantly lower than that in the middle dose group of Xiao Chaihu granules (P ⁇ 0.05, P ⁇ 0.01).

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Abstract

本发明采用肺炎支原体感染的SD大鼠模型进行实验，发现小柴胡颗粒能够清除肺炎支原体感染所致的过量自由基，特别地，小柴胡颗粒与磷酸氯喹联合一起，更能够显著升高血清中谷胱甘肽过氧化物酶、超氧化物歧化酶、过氧化氢酶水平，降低丙二醛含量，表明联合给药可提升机体清除自由基能力，降低自由基堆积产生的细胞毒性，发现小柴胡颗粒联合磷酸氯喹片具有清除肺炎支原体感染所致的过量自由基这一用途，解决过量自由基在肺炎支原体感染中的问题，并对阿奇霉素具有更好的增敏作用，从而可以用于防治肺炎支原体。

Description

小柴胡颗粒联合磷酸氯喹的新应用 技术领域
本发明涉及小柴胡颗粒联合磷酸氯喹的新应用，属于中西药联合用药领域。
背景技术
肺炎支原体是人类原发性非典型肺炎的病原体，也是我国社区获得性肺炎最常见的病原体之一。肺炎支原体感染主要经飞沫传染，会引发咽炎、支气管炎、急性肺炎等上、下呼吸道疾病，同时还可引起脑炎、神经根炎、心肌炎、溶血性贫血等并发症。大环内酯类抗生素是其治疗的首选药物，然而研究表明，我国肺炎支原体在体外对大环内酯类抗生素的耐药率明显高于其他国家，因此应提高警惕，并尝试其他用药组合。最新临床研究表明，用于治疗新型冠状病毒肺炎的洛匹那韦利托那韦(克力芝)和阿比多尔在改善临床症状和加速病毒清除方面均未优于对照组，疗效不显著。这可能是由于肺炎类疾病是一个多系统紊乱的复杂疾病，仅仅通过主动进攻病毒来治疗疾病的效果不好，而开展中西药联合，在抗击感染时结合抗炎、抗病毒或清除自由基的防守性治疗或许可以取得较好的疗效。
小柴胡汤为中医八大名方之一，系医圣张仲景所著《伤寒杂病论》中少阳病的代表性方剂。小柴胡颗粒系广州白云山光华制药依据小柴胡汤首家研制而成的颗粒型中成药制剂，由柴胡、黄芩、半夏(姜制)、党参、生姜、甘草、大枣七个药味按照“君臣佐使”组方而成，具有解表散热，疏肝和胃功效，用于外感病，邪犯少阳证。小柴胡组方已被证明具有解热抗炎、调节免疫、抗肝纤维化、抗过敏等多方面功效。近年来，小柴胡颗粒在治疗感冒、流感等伤寒疫 症方面疗效显著。2003年，小柴胡颗粒被列入广东省抗击“非典”专用药物；2009年，小柴胡颗粒用于抗击全球“甲型流感”；2013年，小柴胡颗粒被广东省卫健委列入登革热诊疗指南救治用药。
磷酸氯喹片用于治疗对氯喹敏感的恶性疟、间日疟及三日疟。此外磷酸氯喹还具有免疫调节活性，并可通过抑制病毒基因表达、改变内吞体pH值、影响自噬反应等机制发挥抗病毒及减轻肺损伤作用。当前，细胞实验表明磷酸氯喹能有效抑制新型冠状病毒感染，已被列入国家卫生健康委员会、国家中医药管理局共同发布的《新型冠状病毒肺炎诊疗方案(试行第六版)》，用于新型冠状病毒感染肺炎的临床治疗。
发明内容
本发明的目的之一是提供一种中药组合物(小柴胡颗粒)的新应用。
具体技术方案如下。
一种中药组合物在制备清除肺炎支原体感染所致的过量自由基的药物中的应用，所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
本发明的另一目的是提供上述中药组合物联合磷酸氯喹的新应用。
具体技术方案如下。
一种中药组合物联合磷酸氯喹在制备清除肺炎支原体感染所致的过量自由基的药物中的应用，所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
本发明的另一目的是提供一种清除肺炎支原体感染所致的过量自由基的联合用药物。
具体技术方案如下。
一种清除肺炎支原体感染所致的过量自由基的联合用药物，包括中药组合物和磷酸氯喹，所述中药组合物与所述磷酸氯喹分别成为独立的给药单元，或所述中药组合物与所述磷酸氯喹共同形成组合的给药单元；所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
本发明另一目的是还提供了所述的联合用药物制备防治肺炎支原体的药物中的应用。
一种中药组合物联合磷酸氯喹在制备防治肺炎支原体的药物中的应用，所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
一种治疗肺炎支原体的联合用药物，包括中药组合物和磷酸氯喹，所述中药组合物与所述磷酸氯喹分别成为独立的给药单元，或所述中药组合物与所述磷酸氯喹共同形成组合的给药单元；所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
在其中一个实施例中，所述中药组合物的活性成分的重量份组成为：柴胡150份、黄芩56份、姜半夏56份、党参56份、生姜56份、甘草56份、大枣56份。
在其中一个实施例中，所述中药组合物是小柴胡颗粒。
在其中一个实施例中，所述磷酸氯喹为片剂。
本发明采用肺炎支原体感染的SD大鼠模型进行实验，发现中药组合物(柴胡、黄芩、姜半夏、党参、生姜、甘草、大枣)能够清除肺炎支原体感染所致的过量自由基，特别地，该中药组合物(例如小柴胡颗粒)与磷酸氯喹联合一起，更能够显著升高血清中谷胱甘肽过氧化物酶、超氧化物歧化酶、过氧化氢酶水平，降低丙二醛含量，且其作用强于小柴胡颗粒与磷酸氯喹单独使用的作用，表明联合给药可提升机体清除自由基能力，降低自由基堆积产生的细胞毒性，发现小柴胡颗粒联合磷酸氯喹片具有清除肺炎支原体感染所致的过量自由基这一用途，解决过量自由基在肺炎支原体感染中的问题。并且发现小柴胡颗粒具有清除肺炎支原体作用且对阿奇霉素具有增敏作用、小柴胡颗粒联合磷酸氯喹片对阿奇霉素具有更好的增敏作用。小柴胡颗粒联合磷酸氯喹可以用于防治肺炎支原体。
附图说明
图1是本发明实施例2各实验组对血清中ROS含量的实验结果柱形图；
图2是本发明实施例2各实验组对血清中GSH-PX含量的实验结果柱形图；
图3是本发明实施例2各实验组对血清中SOD含量的实验结果柱形图；
图4是本发明实施例2各实验组对血清中CAT含量的实验结果柱形图；
图5是本发明实施例2各实验组对血清中MDA含量的实验结果柱形图。
具体实施方式
本发明下列实施例未注明具体条件的实验方法，通常按照常规条件，或按照制造厂商所建议的条件。实施例中所用到的各种常用化学试剂，均为市售产品。
除非另有定义，本发明所使用的所有的技术和科学术语与属于本发明的技 术领域的技术人员通常理解的含义相同。本发明的说明书中所使用的术语只是为了描述具体的实施例的目的，不用于限制本发明。
本发明的术语包括”和“具有”以及它们任何变形，意图在于覆盖不排他的包含。例如包含了一系列步骤的过程、方法、装置、产品或设备没有限定于已列出的步骤或组分，而是可选地还包括没有列出的步骤，或可选地还包括对于这些过程、方法、产品或设备固有的其它步骤或组分。
在本发明中提及的“多个”是指两个或两个以上。“和/或”，描述关联对象的关联关系，表示可以存在三种关系，例如，A和/或B，可以表示：单独存在A，同时存在A和B，单独存在B这三种情况。字符“/”一般表示前后关联对象是一种“或”的关系。
机体在感染后肺炎支原体感染，免疫系统被过度激活，免疫细胞产生大量的细胞因子和自由基，自由基得不到及时清除，作用于机体造成多器官损伤甚至引起器官衰竭。因此，清除过量的自由基对人体器官有一定的保护作用，可减轻肺炎支原体造成的损伤。
本发明在其中一个方面，提供一种中药组合物在制备清除肺炎支原体感染所致的过量自由基的药物中的应用，所述中药组合物的活性成分为柴胡、黄芩、姜半夏、党参、生姜、甘草、大枣。
在其中一个实施例中，柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
在其中一个优选的实施例中，所述中药组合物的活性成分的重量份组成为：柴胡150份、黄芩56份、姜半夏56份、党参56份、生姜56份、甘草56份、大枣56份。在其中一些实施例中，所述所述中药组合物可以是膏剂、汤剂或者浓缩丸、合剂(包括口服液)、糖浆剂、或是颗粒剂等，在一个优选的实施例中， 其为颗粒剂。
进一步地，上述中药组合物联合磷酸氯喹在制备清除肺炎支原体感染所致的过量自由基的药物中的应用。
本发明在另一方面是涉及一种清除肺炎支原体感染所致的过量自由基的联合用药物，包括上述中药组合物和磷酸氯喹，所述中药组合物与所述磷酸氯喹分别成为独立的给药单元，或所述中药组合物与所述磷酸氯喹共同形成组合的给药单元。即所述中药组合物(例如小柴胡颗粒)、磷酸氯喹可以是分开单独给药，也可以是两者组合在一起用药。
磷酸氯喹可以是片剂或者胶囊，或者是其他口服剂型。在一个优选的实施例中，其为片剂。
本发明的一个实施中，还公开了小柴胡颗粒具有清除肺炎支原体作用且对阿奇霉素具有增敏作用、小柴胡颗粒联合磷酸氯喹片对阿奇霉素具有更好的增敏作用。小柴胡颗粒可以用于制备治疗肺炎支原体的药物中，特别是用于辅助阿奇霉素在治疗肺炎支原体的药物中应用。进一步地，小柴胡颗粒联合磷酸氯喹应用于制备肺炎支原体的药物中，在其中一个优选的实施中，小柴胡颗粒联合磷酸氯喹应用于制备辅助阿奇霉素的肺炎支原体的药物中。
为了便于理解本发明，下面将对本发明进行更全面的描述。本发明可以以许多不同的形式来实现，并不限于本文所描述的实施例。相反地，提供这些实施例的目的是使对本发明公开内容的理解更加透彻全面。
以下实施例中的小柴胡颗粒由广州白云山光华制药出售，其活性成分具体组成为：柴胡150g、黄芩56g、姜半夏56g、党参56g、生姜56g、甘草56g、大枣56g，制剂为颗粒剂。
实施例1
本发明通过肺炎支原体标准菌株体外抑菌试验，发现小柴胡颗粒对肺炎支原体标准菌株FH的最小抑菌浓度为3.125mg/mL，磷酸氯喹片对肺炎支原体标准菌株FH的最小抑菌浓度为8mg/mL，小柴胡颗粒联合阿奇霉素可以显著降低阿奇霉素的最小抑菌浓度(0.002mg/ML→0.0005mg/mL)，小柴胡颗粒+磷酸氯喹片联合阿奇霉素可以显著降低阿奇霉素的最小抑菌浓度(0.002mg/ML→0.000125mg/mL)，表明小柴胡颗粒、小柴胡颗粒联合阿奇霉素、小柴胡颗粒+磷酸氯喹片联合阿奇霉素在体外能够显著抑制肺炎支原体，即发现小柴胡颗粒具有清除肺炎支原体作用且对阿奇霉素具有增敏作用、小柴胡颗粒+磷酸氯喹片对阿奇霉素具有增敏作用。
通过以下具体实施例对小柴胡颗粒联合磷酸氯喹片对阿奇霉素抑制肺炎支原体的增敏作用进行进一步说明：
1、肺炎支原体菌株的培养
将肺炎支原体标准菌株MPFH接种至PPLO完全培养液中，放入含CO 2、饱和氮的37℃的生化培养箱中，待培养液由红变黄时开始传代。采用CCU定量测定MPFH浓度，取浓度为1×10 5CCU/mL的菌液备用。
2、药液的准备
小柴胡颗粒：开水冲泡，使其完全溶解，灭菌消毒后冷藏，制成1000mg/mL的小柴胡颗粒药液。
磷酸氯喹片：加水适量，超声震荡，充分溶解，灭菌消毒后冷藏，制成1280mg/mL的磷酸氯喹片药液。
阿奇霉素：在无菌条件下制备成终浓度为1280μg/mL的阿奇霉素药液。
3、最小抑菌浓度(MIC)测定
MIC是考查药物在体外是否具有抑制微生物生长繁殖能力的指标。采用液体微量稀释法在96孔板上测定MIC。使用PPLO培养基(含酚红指示剂)将各药液稀释为10个等倍浓度梯度，接种等量MPFH菌液(1×10 5CCU/mL)，使得各孔液体总体积为200μL，每个样品重复三次。以不加药液孔为阳性对照，以单加培养基孔为阴性对照。滴入无菌液体石蜡密封，盖上盖板，放入37℃培养箱孵育2-3天。观察培养基颜色，对照孔从红变黄时为阳性，颜色不变为阴性。不生长肺炎支原体的最低药液浓度为该药对肺炎支原体的MIC值。小柴胡颗粒联合阿奇霉素给药组加入最小抑菌浓度的小柴胡颗粒，设置10个浓度的阿奇霉素，记录联合给药后阿奇霉素的MIC值；磷酸氯喹片联合阿奇霉素给药组加入最小抑菌浓度的磷酸氯喹片，设置10个浓度的阿奇霉素，记录联合给药后阿奇霉素的MIC值；小柴胡颗粒+磷酸氯喹片联合阿奇霉素给药组加入最小抑菌浓度的小柴胡颗粒、磷酸氯喹片，设置10个浓度的阿奇霉素，记录联合给药后阿奇霉素的MIC值。
3、实验结果
小柴胡颗粒、磷酸氯喹片、阿奇霉素、小柴胡颗粒联合阿奇霉素、磷酸氯喹片联合阿奇霉素、小柴胡颗粒+磷酸氯喹片联合阿奇霉素对MPFH的最小抑菌浓度见表1。结果表明，小柴胡颗粒对肺炎支原体标准菌株FH的最小抑菌浓度为3.125mg/mL，磷酸氯喹片对肺炎支原体标准菌株FH的最小抑菌浓度为8mg/mL，小柴胡颗粒联合阿奇霉素可以显著降低阿奇霉素的最小抑菌浓度(0.002mg/ML→0.0005mg/mL)，小柴胡颗粒+磷酸氯喹片联合阿奇霉素可以显著降低阿奇霉素的最小抑菌浓度(0.002mg/ML→0.000125mg/mL)。
表1 不同药物处理对MPFH的最小抑菌浓度(MIC)
Figure PCTCN2020122715-appb-000001
Figure PCTCN2020122715-appb-000002
实施例2
本发明采用肺炎支原体感染的SD大鼠模型进行实验，发现小柴胡颗粒，特别是小柴胡颗粒联合磷酸氯喹片能够显著升高血清中谷胱甘肽过氧化物酶、超氧化物歧化酶、过氧化氢酶水平，降低丙二醛含量，且其作用强于小柴胡颗粒与磷酸氯喹单独用药，表明联合给药可提升机体清除自由基能力，降低自由基堆积产生的细胞毒性，即发现小柴胡颗粒联合磷酸氯喹片具有清除肺炎支原体感染所致的过量自由基这一用途。
通过以下具体的实施例对小柴胡颗粒联合磷酸氯喹片用于清除肺炎支原体感染所致的过量自由基的用途进行进一步说明：
1、实验动物
SPF级雄性大鼠100只，SD属，体重240-260g，由广东省医学实验动物中心提供，SYXK(粤)2018－0002)。在实验动物适应新环境一周后开始实验，并在实验过程中采取适当的方法减轻对动物的伤害。
2、肺炎支原体菌株的培养
将肺炎支原体标准菌株MPFH接种至PPLO完全培养液中，放入含CO 2、饱和氮的37℃的生化培养箱中，待培养液由红变黄时开始传代。采用CCU定量测定MPFH浓度，取浓度为1×10 8CCU/mL的菌液备用。
3、动物实验分组及给药
SD大鼠随机分为10组，空白对照组、肺炎支原体感染模型组、阳性药阿 奇霉素组(10mg/kg)、磷酸氯喹组(45mg/kg，为人体临床等效剂量)、小柴胡颗粒低剂量组(5.4g/kg，为人体临床等效剂量)、小柴胡颗粒中剂量组(10.8g/kg)、小柴胡颗粒高剂量组(21.6g/kg)、小柴胡颗粒与磷酸氯喹片联合给药低剂量组(小柴胡颗粒5.4g/kg，磷酸氯喹片45mg/kg)、联合给药中剂量组(10.8g/kg，45mg/kg)、联合给药高剂量组(21.6g/kg，45mg/kg)，每组10只。采用乙醚轻度麻醉大鼠，除空白对照组外，其余各组大鼠滴鼻接种1×10 8CCU/mL的MPFH菌液，每天100μL，共4天，空白对照组滴鼻等量生理盐水。各组大鼠在最后一次接种的第2天开始干预，阿奇霉素采用皮下注射给药，小柴胡颗粒组与联合给药组采用灌胃给药，空白对照组和模型组以等量生理盐水灌胃，每天给药1次，连续给药7天。模型建立后，各组动物分笼饲养，实验组和空白对照组隔离饲养。
4、样品采集及药效指标检测
末次给药1h后，各组大鼠采用乙醚麻醉，心脏取血至普通采血管，室温放置30min，5000r/min离心5min，得血清。试剂盒检测血清中活性氧(ROS)、谷胱甘肽过氧化物酶(GSH-PX)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、丙二醛(MDA)的含量。
5、实验结果
已有研究表明，肺炎支原体感染的患者体内存在大量自由基，这些自由基的急剧堆积诱导免疫细胞产生大量的促炎细胞因子，加速了疾病的发生和发展。ROS是指机体内或者自然环境中由氧组成,含氧并且性质活泼的物质的总称，包括超氧阴离子自由基、羟自由基、氢过氧自由基以及过氧化氢、过氧化脂质等。ROS可氧化蛋白质、DNA和脂质，诱发细胞凋亡，直接参与多种肺部疾病的病理过程，导致肺部损伤并影响肺泡修复功能。GSH-PX能够清除细胞中的过 氧化物和羟自由基，阻断脂质过氧化连锁反应，可以起到保护细胞结构的作用，是体内广泛存在的一种自由基清除酶。SOD是机体内主要的抗氧化酶，能够催化超氧阴离子自由基歧化生成氧和过氧化氢，保护细胞免受损害。CAT是催化过氧化氢分解成氧和水的酶，是过氧化氢自由基清除剂。MDA含量则能够间接反映机体细胞受自由基攻击的严重程度。
实验结果见图1-图5，其中各数字对应的实验组为：1-空白对照组；2-肺炎支原体感染模型组；3-阳性药阿奇霉素组；4-磷酸氯喹组；5-小柴胡颗粒低剂量组；6-小柴胡颗粒中剂量组；7-小柴胡颗粒高剂量组；8-联合给药低剂量组；9-联合给药中剂量组；10-联合给药高剂量组。a代表P<0.01vs空白组；b、c代表P＜0.05和P＜0.01vs模型组；d、e代表P＜0.05和P＜0.01联合给药组vs磷酸氯喹组；f、g代表P＜0.05和P＜0.01联合给药组vs小柴胡颗粒组(联合给药低剂量组vs小柴胡颗粒低剂量组，联合给药中剂量组vs小柴胡颗粒中剂量组，联合给药高剂量组vs小柴胡颗粒高剂量组)(n＝10)。
实验结果(表2、图1、图2、图3、图4、图5)表明，空白对照组与模型组大鼠的ROS、GSH-PX、SOD、CAT、MDA含量均存在显著差异(P＜0.01)，其原因可能是机体感染支原体后，过氧化反应增加，自由基清除酶被过度消耗，组织中的活性氧堆积，产生细胞毒性。与模型组相比，小柴胡颗粒、磷酸氯喹片、小柴胡颗粒和磷酸氯喹片联合给药低、中、高剂量均可显著升高抗氧化物酶GSH-PX、SOD和CAT水平，降低ROS、MDA含量(P＜0.05，P＜0.01)。与磷酸氯喹组相比，联合给药低、中、高剂量组的GSH-PX、SOD水平显著升高，ROS含量显著降低(P＜0.05，P＜0.01)，中、高剂量组的CAT、MDA含量与磷酸氯喹组有显著差异(P＜0.05，P＜0.01)。与小柴胡颗粒组相比，联合给药组低、中、高剂量组的GSH-PX水平显著高于小柴胡颗粒对应剂量组，ROS 含量显著低于小柴胡颗粒对应剂量组；联合给药组中、高剂量组的SOD、CAT水平与小柴胡颗粒对应剂量组存在显著差异；联合给药组中剂量组的MDA含量显著低于小柴胡颗粒中剂量组(P＜0.05，P＜0.01)。
实验结果表明磷酸氯喹、小柴胡颗粒以及小柴胡颗粒联合磷酸氯喹片均可提高机体清除自由基能力，抑制过氧化反应，减轻机体的氧化应急损伤。同时小柴胡颗粒联合磷酸氯喹片的作用整体优于小柴胡颗粒或磷酸氯喹的作用，表明二者联用在清除自由基方面的效果更优。
表2 小柴胡颗粒联合磷酸氯喹片对血清ROS、GSH-PX、SOD、CAT、MDA的影响(n＝10)
Figure PCTCN2020122715-appb-000003
以上所述实施例仅表达了本发明的几种实施方式，其描述较为具体和详细但并不能因此而理解为对发明专利范围的限制。应当指出的是，对于本领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干变形和改进，这些都属于本发明的保护范围。因此，本发明专利的保护范围应以所附权利要求为准。

Claims (10)

  1. 一种中药组合物在制备清除肺炎支原体感染所致的过量自由基的药物中的应用，所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
  2. 根据权利要求1所述的应用，其特征在于，所述中药组合物是小柴胡颗粒；和/或所述中药组合物的活性成分的重量份组成为：柴胡150份、黄芩56份、姜半夏56份、党参56份、生姜56份、甘草56份、大枣56份。
  3. 一种中药组合物联合磷酸氯喹在制备清除肺炎支原体感染所致的过量自由基的药物中的应用，所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
  4. 根据权利要求3所述的应用，其特征在于，所述磷酸氯喹为片剂。
  5. 根据权利要求3或4所述的应用，其特征在于，所述中药组合物是小柴胡颗粒；和/或所述中药组合物的活性成分的重量份组成为：柴胡150份、黄芩56份、姜半夏56份、党参56份、生姜56份、甘草56份、大枣56份。
  6. 一种清除肺炎支原体感染所致的过量自由基的联合用药物，其特征在于，包括中药组合物和磷酸氯喹，所述中药组合物与所述磷酸氯喹分别成为独立的给药单元，或所述中药组合物与所述磷酸氯喹共同形成组合的给药单元；所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
  7. 根据权利要求6所述的联合用药物，其特征在于，所述磷酸氯喹为片剂。
  8. 根据权利要求6或7所述的联合用药物，其特征在于，所述中药组合物 为小柴胡颗粒；和/或所述中药组合物的活性成分的重量份组成为：柴胡150份、黄芩56份、姜半夏56份、党参56份、生姜56份、甘草56份、大枣56份。
  9. 根据权利要求6-8任一项所述的联合用药物制备防治肺炎支原体的药物中的应用。
  10. 一种中药组合物在制备防治肺炎支原体的药物中的应用，所述中药组合物的活性成分的重量份组成为：柴胡150±5份、黄芩56±5份、姜半夏56±3份、党参56±3份、生姜56±3份、甘草56±3份、大枣56±3份。
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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103954723A (zh) * 2014-04-28 2014-07-30 四川逢春制药有限公司 小柴胡颗粒的检测方法
CN103961665A (zh) * 2014-05-07 2014-08-06 江西百神药业股份有限公司 一种小柴胡颗粒的制备方法
CN108424434A (zh) * 2017-02-13 2018-08-21 四川大学 氯喹胆固醇衍生物及其制备方法和用途
CN111329982A (zh) * 2020-03-12 2020-06-26 广州白云山光华制药股份有限公司 小柴胡颗粒联合磷酸氯喹的新应用

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8440695B2 (en) * 2005-11-09 2013-05-14 St Jude Children's Research Hospital Use of chloroquine to treat metabolic syndrome
CN102349993B (zh) * 2011-10-18 2012-10-03 广州白云山光华制药股份有限公司 一种小柴胡颗粒剂及其制备方法
CN105106914B (zh) * 2015-09-25 2019-03-12 山西医科大学 一种小柴胡缓释片剂及其制备方法
CN109125699A (zh) * 2018-10-24 2019-01-04 遵义医学院 小柴胡汤在制备治疗肝性脑病药物中的应用

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103954723A (zh) * 2014-04-28 2014-07-30 四川逢春制药有限公司 小柴胡颗粒的检测方法
CN103961665A (zh) * 2014-05-07 2014-08-06 江西百神药业股份有限公司 一种小柴胡颗粒的制备方法
CN108424434A (zh) * 2017-02-13 2018-08-21 四川大学 氯喹胆固醇衍生物及其制备方法和用途
CN111329982A (zh) * 2020-03-12 2020-06-26 广州白云山光华制药股份有限公司 小柴胡颗粒联合磷酸氯喹的新应用

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
JIANG HAILI: "Xiaochaihu and Shigao Decoction in the Treatment of Infantile Mycoplasmal Pneumonia for 36 Cases", CHINESE MEDICINE MODERN DISTANCE EDUCATION OF CHINA, vol. 13, no. 14, 1 January 2015 (2015-01-01), pages 53 - 54, XP055852614, ISSN: 1672-2779, DOI: 10.3969/j.issn.1672-2779.2015.14.028 *
LIANG XUE: "Research on the Treatment of Mycoplasma Pneumonia in Children Using Minor Bupleurum Decoction (Xiao Chai Hu Tang) and Gypsum Decoction (Shi Gao Tang)", WORLD LATEST MEDICINE INFORMATION, vol. 17, no. 91, 14 November 2017 (2017-11-14), pages 123, XP055852703, DOI: 10.19613/j.cnki.1671-3141.2017.91.100 *
SHIMIZU TAKASHI, KIMURA YUI, KIDA YUTAKA, KUWANO KOICHI, TACHIBANA MASATO, HASHINO MASANORI, WATARAI MASAHISA: "Cytadherence of Mycoplasma pneumoniae Induces Inflammatory Responses through Autophagy and Toll-Like Receptor 4", INFECTION AND IMMUNITY, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 82, no. 7, 1 July 2014 (2014-07-01), US, pages 3076 - 3086, XP055852708, ISSN: 0019-9567, DOI: 10.1128/IAI.01961-14 *
SUN LEI: "Treating 40 Cases of Mycoplasma Pneumonia with Modified Minor Bupleurum Decoction (Xiao Chai Hu Tang))", DIET HEALTH, vol. 6, no. 40, 19 August 2019 (2019-08-19), pages 61, XP009530761, ISSN: 2095-8439 *
XIAO CHAI, HU TANG: " A Case of Treating Children with Atelectasis Caused by Mycoplasma Pneumonia Using Modified Minor Bupleurum Decoction ", PROCEEDINGS OF THE 2ND NATIONAL ACADEMIC CONFERENCE ON PREVENTION AND TREATMENT OF RESPIRATORY DISEASES WITH INTEGRATED TRADITIONAL AND WESTERN MEDICINE, 15 March 2013 (2013-03-15), pages 600 - 602, XP009530762 *

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