Antibacterial pharmaceutical composition containing meconopsis horridis

Abstract

The invention provides a pharmaceutical composition containing meconopsis horridis, which is prepared from the following raw materials in parts by weight: meconopsis horridis ethanol extract, stigma croci ethanol extract and myrobalan ethanol extract, and the extracts are calculated by dry weight. The invention also provides application of the pharmaceutical composition in preparation of antibacterial drugs.

Description

Technical Field

[0001] The invention relates to the field of Tibetan antibacterial drugs, and in particular to antibacterial drugs containing Tibetan medicines such as Meconopsis spinosa. Background Art

[0002] Bacterial infection refers to a series of pathological changes caused by bacteria invading the host, growing and multiplying and interacting with the body. Clinically, bacteria that can cause host infection are called pathogenic bacteria or pathogens. Some bacteria are not pathogenic under normal circumstances, but can cause diseases under specific conditions such as decreased host immune defense or dysbiosis. Such bacteria are called opportunistic pathogens or conditional pathogens. After the human body is infected with bacteria, a series of symptoms may occur, such as inflammation, rash, fever, sepsis, etc.

[0003] Klebsiella pneumoniae is the most important type of bacteria in the genus Klebsiella of the family Enterobacteriaceae (commonly known as pneumonia bacillus), and the diseases caused by it account for more than 95% of Klebsiella infections. It exists in the upper respiratory tract and intestines of the human body. When the body's resistance is reduced, it enters the lungs through the respiratory tract and causes large or small lobular consolidation, which is more common in the upper lobes. Klebsiella pneumoniae is a Gram-negative bacillus. The exudate in the lesion is viscous and heavy, causing the interlobular space to fall. The bacteria have capsules. When they grow and multiply in the alveoli, they cause tissue necrosis, liquefaction, and the formation of single or multiple abscesses. When the lesion involves the pleura and pericardium, it can cause exudative or purulent effusion. The fibrous tissue in the lesion proliferates actively and is easy to organize; fibrinous pleural effusions may have adhesions in the early stage. In hospital-acquired sepsis, Klebsiella, Pseudomonas aeruginosa, and Serratia are all important pathogens with a high mortality rate.

[0004] Pseudomonas aeruginosa is an obligate aerobic bacterium that exists in water, soil, plants, animals, and even humans. During its growth, it produces water-soluble pigments pyocyanin and pyocyanin with fluorescent pigments, which make the pus or dressings after infection appear green, hence the name. When the body's immunity is low, infection can occur in various parts of the body, including burns or trauma sites, urinary tract, cornea, gastrointestinal tract, etc., and can cause otitis media, keratitis, urethritis, gastroenteritis, endocarditis, and empyema. In addition, bacteremia, sepsis, and severe epidemic diarrhea in infants caused by this bacterium have also been reported.

[0005] Currently, antibiotics are generally used to treat bacterial infections. However, the use of antibiotics has led to the emergence of a large number of clinical drug-resistant bacteria. Summary of the invention

[0006] The present invention intends to select some drugs from Tibetan medicine and compound them, so as to find a drug composition that can effectively fight bacteria, especially Klebsiella pneumoniae and Pseudomonas aeruginosa, to provide a new treatment for such bacterial infections, thereby avoiding the use of antibiotics and increasing the chance of drug resistance.

[0007] The invention provides a pharmaceutical composition containing Meconopsis spinulosa, wherein the raw materials are selected from the following components in parts by weight: Meconopsis spinulosa ethanol extract, saffron ethanol extract and terminalia chebula ethanol extract, and the extracts are calculated by dry weight.

[0008] The weight portion of the ethanol extract of Meconopsis spinulosa is 5 to 15 parts, further selected from 8 to 12 parts, and further selected from 10 to 11 parts. The weight portion of the ethanol extract of Meconopsis spinulosa includes but is not limited to 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, etc.

[0009] The weight portion of the ethanol extract of Terminalia chebula is 1 to 10 parts, further selected from 1 to 8 parts, further selected from 1 to 7 parts, and can also be selected from 1 to 2 parts or 6 to 7 parts. The weight portions of the ethanol extract of Terminalia chebula include but are not limited to 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, etc.

[0010] Wherein, the weight portion of saffron ethanol extract is 1 to 60 parts, further selected from 1 to 50 parts. The weight portions of saffron ethanol extract include but are not limited to 1 part, 2 parts, 3 parts, 4 parts, 5 parts, 6 parts, 7 parts, 8 parts, 9 parts, 10 parts, 11 parts, 12 parts, 13 parts, 14 parts, 15 parts, 16 parts, 17 parts, 18 parts, 19 parts, 20 parts, 21 parts, 22 parts, 23 parts, 24 parts, 25 parts, 26 parts, 27 parts, 28 parts, 29 parts, 30 parts, 31 parts, 32 parts, 33 parts, 34 parts, 35 parts, 36 parts, 37 parts, 38 parts, 39 parts, 40 parts, 41 parts, 42 parts, 43 parts, 44 parts, 45 parts, 46 parts, 47 parts, 48 ​​parts, 49 parts, 50 parts, etc.

[0011] Furthermore, Meconopsis spinulosa ethanol extract: Terminalia chebula ethanol extract: Crocus sativus ethanol extract = (10-11): (1-7): (48-49).

[0012] Furthermore, the ethanol extract of Meconopsis spinulosa: the ethanol extract of Terminalia chebula: the ethanol extract of Crocus sativus = (10-11): (1-2): (48-49); or, the ethanol extract of Meconopsis spinulosa: the ethanol extract of Terminalia chebula: the ethanol extract of Crocus sativus = (10-11): (6-7): (48-49).

[0013] In certain specific embodiments of the present invention, when the composition is liquid, the ethanol extract of Meconopsis spinulosa is 10-11 mg / ml, the ethanol extract of Terminalia chebula is 1-2 mg / ml, and the ethanol extract of Crocus sativus is 48-49 mg / ml; or, the ethanol extract of Meconopsis spinulosa is 10-11 mg / ml, the ethanol extract of Terminalia chebula is 6-7 mg / ml, and the ethanol extract of Crocus sativus is 48-49 mg / ml.

[0014] In certain specific embodiments of the present invention, the ethanol extract is an ethanol extract with a concentration of more than 80% V / V.

[0015] In certain specific embodiments of the present invention, the ethanol extract is an ethanol extract with a concentration of more than 95% V / V.

[0016] In certain embodiments of the present invention, the ethanol extract is a 95% V / V ethanol or anhydrous ethanol extract.

[0017] The present invention also provides a method for preparing the above-mentioned pharmaceutical composition, comprising the following contents:

[0018] (1) extracting each raw material with ethanol of corresponding concentration, concentrating and drying the extract to obtain each ethanol extract;

[0019] (2) Mix the ethanol extracts according to the aforementioned weight portions to obtain the product.

[0020] The present invention also provides use of the above-mentioned pharmaceutical composition in preparing antibacterial drugs.

[0021] Furthermore, the drug is a drug against Klebsiella pneumoniae and / or Pseudomonas aeruginosa.

[0022] Klebsiella pneumoniae is the most important type of bacteria in the genus Klebsiella of the family Enterobacteriaceae (commonly known as pneumonia bacillus), and the diseases caused by it account for more than 95% of Klebsiella infections. It exists in the upper respiratory tract and intestines of the human body. When the body's resistance is reduced, it enters the lungs through the respiratory tract and causes large or small lobular consolidation, which is more common in the upper lobes. When it grows and reproduces in the alveoli, it causes tissue necrosis, liquefaction, and the formation of single or multiple abscesses. When the lesion involves the pleura and pericardium, it can cause exudative or purulent effusion. The fibrous tissue in the lesion proliferates actively and is easy to organize; fibrinous pleural effusions may show adhesions in the early stage. In hospital-acquired sepsis, Klebsiella is an important pathogen with a high mortality rate.

[0023] Pseudomonas aeruginosa, also known as Pseudomonas aeruginosa, is a Gram-negative, aerobic, long rod-shaped bacterium with only unidirectional motility. It is an opportunistic infection bacterium and also an opportunistic infection for plants. After infection, the pus and exudate are green. Pseudomonas aeruginosa can produce a variety of substances related to toxicity, such as endotoxins, exotoxin a, elastase, collagenase, pancreatic peptidase, etc., among which exotoxin a is the most important. Pseudomonas aeruginosa exotoxin a is a heat-labile single-chain polypeptide with a molecular weight of about 66,000. It can be detoxified into a toxoid by treatment with formaldehyde or glutaraldehyde and neutralized by specific antitoxins. It is highly toxic. After injection into animals, cell swelling, fatty degeneration and necrosis may occur in the main target organ liver; other organ lesions include pulmonary hemorrhage and renal necrosis. The mechanism of exotoxin a is somewhat similar to that of diphtheria toxin, that is, it ultimately inactivates elongation factor 2 (ef~2) on the ribosome and inhibits protein synthesis in host cells, but the specific process is different. In addition, exotoxin a and diphtheria toxin differ in protein structure, immunogenicity, whip cells and sensitive animals. Pseudomonas aeruginosa infection can occur in any part and tissue of the human body, commonly in burns or trauma sites, middle ear, cornea, urethra and respiratory tract. It can also cause endocarditis, gastroenteritis, empyema and even sepsis. After infection, patients can produce specific antibodies, which have a certain anti-infection effect. The use of anti-Pseudomonas aeruginosa immune serum can reduce the incidence and mortality of secondary sepsis in patients.

[0024] Klebsiella pneumoniae and Pseudomonas aeruginosa are common drug-resistant bacteria in clinical practice. The present invention further found that the pharmaceutical composition of the present invention also has good inhibitory activity against such drug-resistant bacteria. Based on such research results, in the present invention, the drug is a drug against drug-resistant bacteria.

[0025] Furthermore, the drug-resistant bacteria are selected from:

[0026] Klebsiella pneumoniae that is resistant to one or more of the following drugs: doxycycline, neomycin, tetracycline, oxytetracycline, ampicillin, penicillin, clindamycin, lincomycin, sulfamethoxazole, midecamycin, kanamycin;

[0027] or / and, Pseudomonas aeruginosa resistant to one or more of the following drugs: cefotaxime, ceftriaxone, cefoperazone, polymyxin, mezlocillin, amoxicillin, tylosin, tetracycline, doxycycline, rifampicin, trimethoprim, tylosin.

[0028] Furthermore, the medicine of the present invention is used to treat infections caused by the bacteria.

[0029] Meconopsis horridula Hook.f.et Thoms. is an annual herbaceous plant of the genus Meconopsis in the family Papaveraceae. It tastes bitter, is cold in nature, and is slightly toxic. It has the functions of relieving fever, relieving pain, setting bones, and promoting blood circulation and removing blood stasis. It is used to treat head injuries, fractures, bruises, etc. According to the "Jingzhu Materia Medica", "It is warm and bitter in taste, can clear the heat in the bones, and is the nectar for treating skull fractures."

[0030] Saffron is the dried stigma of Crocus sativus L, a plant of the Iridaceae family. It has the effects of promoting blood circulation and removing blood stasis, cooling blood and detoxifying, relieving depression and calming the mind. It is used for amenorrhea, postpartum blood stasis, fever and rashes, depression, palpitations and madness.

[0031] Terminalia chebula Retz. or Terminalia chebula Retz. var. tomentella Kurt. of the Combretaceae family is a dry, mature fruit. It has the effects of astringing the intestines and stopping diarrhea, astringing the lungs and stopping coughs, lowering fire and relieving sore throats. It is often used for chronic diarrhea, dysentery, rectal prolapse due to blood in the stool, asthma and cough due to lung deficiency, chronic cough, sore throat and hoarseness. DETAILED DESCRIPTION

[0032] The raw materials of the medicine of the present invention are purchased from the market.

[0033] The Pseudomonas aeruginosa and Klebsiella pneumoniae used in the present invention are separated from clinical animals by the Veterinary Research Institute of Sichuan Academy of Animal Husbandry Sciences and are stored in the Veterinary Research Institute of Sichuan Academy of Animal Husbandry Sciences.

[0034] Example 1

[0035] (1) Raw materials: Meconopsis pilosa, Saffron, Terminalia chebula

[0036] (2) Weigh each raw material separately for extraction: 90 g of the raw material is crushed, wrapped with filter paper, and then extracted with anhydrous ethanol for 6 h. The ethanol extract is concentrated and dried to obtain the ethanol extract of each raw material.

[0037] Among them, the yield of Meconopsis spinosa ethanol extract was 8.73%, the yield of Crocus saffron ethanol extract was 46.3%, and the yield of Terminalia chebula ethanol extract was 33.93%.

[0038] Example 2

[0039] (1) Raw materials: Meconopsis pilosa, Saffron, Terminalia chebula

[0040] (2) Weigh each raw material separately for extraction: 200 g of the raw material is crushed, wrapped with filter paper, and then extracted with 95% ethanol for 8 h. The ethanol extract is concentrated and dried to obtain the ethanol extract of each raw material.

[0041] Example 3

[0042] (1) Raw materials: Meconopsis pilosa, Saffron, Terminalia chebula

[0043] (2) Weigh each raw material separately for extraction: 500 g of the raw material is crushed, wrapped with filter paper, added with anhydrous ethanol, and refluxed for 5 h. The ethanol extract is collected, concentrated, and dried to obtain the ethanol extract of each raw material.

[0044] Example 4

[0045] (1) Raw materials: Meconopsis pilosa, Saffron, Terminalia chebula

[0046] (2) Weigh each raw material separately for extraction: 500 g of the raw material is crushed, wrapped with filter paper, added with 80% ethanol, and ultrasonically extracted for 2 h. The ethanol extract is collected, concentrated, and dried to obtain the ethanol extract of each raw material.

[0047] Take the extract prepared in Example 1, dilute to obtain 0.69 g / ml of Meconopsis spinosa mother solution, 0.77 g / ml of Crocus sativus mother solution, and 0.87 g / ml of Terminalia chebula mother solution, and set aside.

[0048] Test Example 1 Drug sensitivity test of Pseudomonas aeruginosa

[0049] 1 Materials and methods

[0050] 1.1 Main reagents and instruments

[0051] Tryptic soy agar (TSA) and tryptic soy broth (TSB) were purchased from Oboxin Co., Ltd.; drug sensitivity tablets were purchased from OXOID Company, UK; constant temperature shaking incubator (THZ-92C) was purchased from Changzhou Gode Instrument Manufacturing Co., Ltd.; constant temperature incubator (GH-600) was purchased from Beijing ZTE Weiye Instrument Co., Ltd.

[0052] 1.2 Strain recovery

[0053] Under the clean bench, use a sterile inoculation loop to dip into the preserved bacterial liquid of Pseudomonas aeruginosa, inoculate it on TSA medium by zoning and streaking method, incubate it upside down in a constant temperature incubator at 37℃ for 16h to obtain a single colony, then pick up the single colony with a sterile inoculation loop, inoculate it into a test tube containing 5mL TSB medium, and place it in a constant temperature air oscillator at a speed of 160r / min and 37℃ for 12h of shaking culture.

[0054] 1.3 Drug sensitivity test

[0055] The KB paper method was used to test the drug sensitivity of the strain. Dip the above-mentioned resuscitated strain solution with a sterile cotton swab, squeeze out the excess water on the cotton swab on the inner side of the tube wall, and evenly apply it to the entire surface of the TSA culture medium. After the bacterial solution is completely absorbed, use sterile tweezers to clamp the drug sensitivity paper and stick it on the surface of the plate. The plate was inverted and incubated in a 37°C constant temperature incubator for 16 hours, and the diameter of the inhibition zone was measured. The results were judged according to the antimicrobial drug sensitivity test execution standards issued by the Clinical Laboratory Standards Institute (CLSI) of the United States.

[0056] 2 Results

[0057] The results of drug sensitivity test are shown in Table 1. The strain was sensitive to six drugs including amikacin, tobramycin, levofloxacin, ciprofloxacin, florfenicol and azithromycin, intermediate to two drugs including enrofloxacin and aztreonam, and resistant to twelve drugs including cefotaxime, ceftriaxone, cefoperazone, polymyxin, mezlocillin, amoxicillin, tylosin, tetracycline, doxycycline, rifampicin, trimethoprim and tylosin.

[0058] Table 1 Results of drug sensitivity test of Pseudomonas aeruginosa

[0059]

[0060]

[0061] Note: S indicates drug sensitivity, I indicates drug resistance, and R indicates drug resistance

[0062] Test Example 2 Drug sensitivity test of Klebsiella pneumoniae

[0063] Referring to Test Example 1, a drug resistance study was conducted on Klebsiella pneumoniae. See Table 2 for details.

[0064] Table 2 Results of drug sensitivity test of Klebsiella pneumoniae

[0065]

[0066]

[0067] Note: S indicates drug sensitivity, I indicates drug resistance, and R indicates drug resistance

[0068] Test Example 3 Antibacterial Activity Test of the Pharmaceutical Composition of the Present Invention

[0069] 1. Preparation of bacterial solution

[0070] The culture medium used was Mueller-Hinton (MH) broth recommended by NCCLS, pH 7.2-7.4. Use an inoculation loop to pick up three activated Pseudomonas aeruginosa and Klebsiella pneumoniae, inoculate them into 4 ml of culture medium, and culture them at 35°C until the bacterial solution reaches the logarithmic growth phase. Use the culture medium to calibrate the concentration to 0.5 McFarland turbidimetric standard, and dilute the bacterial suspension 1:100 for later use.

[0071] 2. Drug sensitivity test

[0072] 2.1 Single extract MIC determination:

[0073] The drug sensitivity plate was prepared by the 48-well plate micro-liquid dilution method. The single extracts were Meconopsis spinosa, Terminalia chebula, and saffron alcohol extracts. The first well was added with 200ul of culture medium and 200ul of extract mother solution, and then diluted in multiples. The last well was the control group without adding drug solution. 200ul of bacterial suspension was added to each well.

[0074] 2.2 Drug synergy experiment:

[0075] Group 1, Meconopsis spinulosa and Terminalia chebula group:

[0076] First, add 100ul of culture solution to each well of the culture plate, add 100ul of 4-fold diluted Terminalia chebula solution to the first column, and dilute it to the 8th well; add 100ul of 4-fold diluted Meconopsis pilosa to each well of the 1st to 8th well of the first row, add 100ul of 8-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the second row, add 100ul of 16-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the third row, add 100ul of 32-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the fourth row, and finally add 200ul of prepared Klebsiella pneumoniae suspension to each well. The 12th column was used as a control.

[0077] Group 2, Meconopsis spinulosa, Terminalia chebula and Saffron group (Saffron diluted 16 times):

[0078] First, add 100ul of culture solution to each well of the culture plate, add 100ul of 4-fold diluted Terminalia chebula solution to the first column, and dilute it to the 8th well; add 100ul of 4-fold diluted Meconopsis pilosa to each well of the 1st to 8th well of the first row, add 100ul of 8-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the second row, add 100ul of 16-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the third row, add 100ul of 32-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the fourth row, and finally add 200ul of Klebsiella pneumoniae suspension containing 16-fold diluted saffron solution to each well. The 12th column was used as a control.

[0079] Group 3: Meconopsis spinulosa, Terminalia chebula and saffron group (saffron diluted 32 times):

[0080] First, add 100ul of culture solution to each well of the culture plate, add 100ul of 4-fold diluted Terminalia chebula solution to the first column, and dilute it to the 8th well; add 100ul of 4-fold diluted Meconopsis pilosa to each well of the 1st to 8th well of the first row, add 100ul of 8-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the second row, add 100ul of 16-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the third row, add 100ul of 32-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the fourth row, and finally add 200ul of Klebsiella pneumoniae suspension containing 32-fold diluted saffron solution to each well. The 12th column was used as a control.

[0081] Control group: The control group included culture medium, 16-fold diluted saffron suspension, 32-fold diluted saffron suspension, and 128-fold diluted saffron suspension.

[0082] Group 4, Meconopsis spinulosa and Terminalia chebula group:

[0083] First, add 100ul of culture solution to each well of the culture plate, add 100ul of 4-fold diluted Terminalia chebula solution to the first column, and dilute it to the 8th well; add 100ul of 4-fold diluted Meconopsis pilosa to each well of the 1st to 8th well of the first row, add 100ul of 8-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the second row, add 100ul of 16-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the third row, add 100ul of 32-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the fourth row, and finally add 200ul of prepared Pseudomonas aeruginosa suspension to each well. The 12th column was used as a control.

[0084] Group 5, Meconopsis spinulosa, Terminalia chebula and Saffron group (Saffron diluted 16 times):

[0085] First, add 100ul of culture solution to each well of the culture plate, add 100ul of 4-fold diluted Terminalia chebula solution to the first column, and dilute it to the 8th well; add 100ul of 4-fold diluted Meconopsis pilosa to each well of the 1st to 8th well of the first row, add 100ul of 8-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the second row, add 100ul of 16-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the third row, add 100ul of 32-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the fourth row, and finally add 200ul of Pseudomonas aeruginosa suspension containing 16-fold diluted saffron solution to each well. The 12th column was used as a control.

[0086] Group 6: Meconopsis spinulosa, Terminalia chebula and saffron group (saffron diluted 32 times):

[0087] First, add 100ul of culture solution to each well of the culture plate, add 100ul of 4-fold diluted Terminalia chebula solution to the first column, and dilute it to the 8th well; add 100ul of 4-fold diluted Meconopsis pilosa to each well of the 1st to 8th well of the first row, add 100ul of 8-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the second row, add 100ul of 16-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the third row, add 100ul of 32-fold diluted Meconopsis pilosa to each well of the 1st to 11th well of the fourth row, and finally add 200ul of Pseudomonas aeruginosa suspension containing 32-fold diluted saffron solution to each well. The 12th column was used as a control.

[0088] 3. Determination of minimum inhibitory concentration (MIC value)

[0089] The results were determined after incubation at 35°C for 20 h. All tests were performed twice. The MIC values ​​of the same strain were considered to be the same when the dilutions in the two tests did not exceed 2.

[0090] 4. Minimum bactericidal concentration (MBC) determination

[0091] Aseptically take 3ul of the liquid from the MIC test culture well, inoculate it into solid culture medium, and culture it at 35℃ for 24 hours. Observe the bacterial growth and determine the minimum bactericidal concentration.

[0092] 5. Evaluation of the effect of combined medication

[0093] The fractional inhibitory concentration index (FICI) is used to express it, that is, the MIC value of the drug in combination divided by the sum of the quotients of the MIC value of the drug when used alone, expressed as: FICI = MIC A combination / MIC A alone + MIC B combination / MIC B alone.

[0094] Standard: FICI < 1 indicates synergistic effect, FICI = 1 indicates additive effect, 1 < FICI ≤ 2 indicates irrelevant effect, and FICI > 2 indicates antagonistic effect.

[0095] Table 3 Test results of antibacterial activity of each raw material extract MIC: clear "-", turbid "+"; MBC: no bacteria "N", bacteria "Y"

[0096]

[0097]

[0098] Results: The minimum MIC concentration of Terminalia chebula against Klebsiella pneumoniae was 27.1875 mg / ml, and the minimum MIC concentration against Pseudomonas aeruginosa was 27.1875 mg / ml; the minimum MIC concentration of Meconopsis spinulosa against Klebsiella pneumoniae was 43.125 mg / ml, and the minimum MIC concentration against Pseudomonas aeruginosa was 43.125 mg / ml; Saffron extract had no obvious antibacterial effect on Klebsiella pneumoniae and Pseudomonas aeruginosa.

[0099] Table 4 Results of drug sensitivity and synergy test of Klebsiella pneumoniae MIC: clear "-", turbid "+"; MBC: no bacteria "N", bacteria "Y"

[0100]

[0101]

[0102] Results: The Meconopsis spinulosa and Terminalia chebula contained 21.5625 mg / ml and 6.7969 mg / ml, respectively, in formulation 1, which had an antibacterial effect on Klebsiella pneumoniae. The Meconopsis spinulosa and Terminalia chebula contained 10.7813 mg / ml and 6.7969 mg / ml and 48.125 mg / ml, respectively, in formulation 2, which had an antibacterial effect on Klebsiella pneumoniae.

[0103] Table 5 Results of drug sensitivity and synergistic test of Pseudomonas aeruginosa MIC: clear "-", turbid "+"; MBC: no bacteria "N", bacteria "Y"

[0104]

[0105]

[0106] Results: The Meconopsis spinulosa and Terminalia chebula in formulation 4 contained 21.5625 mg / ml and 3.3984 mg / ml, which had an antibacterial effect on Pseudomonas aeruginosa. The Meconopsis spinulosa, Terminalia chebula and Terminalia chebula in formulation 5 contained 10.7813 mg / ml, 1.6992 mg / ml and 48.125 mg / ml of saffron, which had an antibacterial effect on Klebsiella pneumoniae.

[0107] It can be seen from the above experiment that when saffron 48.125 mg / ml is used alone, it has no inhibitory effect on clinically extracted drug-resistant bacteria. However, when saffron is used in combination with Meconopsis pricklyi and Terminalia chebula, the composition can also exert obvious antibacterial activity at a concentration far below MIC, and FICI is <1, indicating that the present invention uses saffron in combination with Meconopsis pricklyi and Terminalia chebula to produce a synergistic effect.

Claims

1. A pharmaceutical composition containing Meconopsis spinosa, characterized in that: The raw materials are selected from the following components in weight: Meconopsis spinulosa ethanol extract, saffron ethanol extract, terminalia chebula ethanol extract, the extracts are calculated by dry weight; the ethanol extract is an ethanol extract with a concentration of more than 80% V / V.

2. The pharmaceutical composition according to claim 1, characterized in that Meconopsis spinulosa ethanol extract: Terminalia chebula ethanol extract: Crocus sativus ethanol extract = (10-11): (1-7): (48-49).

3. The pharmaceutical composition according to claim 1, characterized in that Meconopsis spinulosa ethanol extract: Terminalia chebula ethanol extract: Crocus sativus ethanol extract = (10-11): (1-2): (48-49); or, Meconopsis spinulosa ethanol extract: Terminalia chebula ethanol extract: Crocus sativus ethanol extract = (10-11): (6-7): (48-49).

4. The pharmaceutical composition according to claim 1, characterized in that When the composition is liquid, the ethanol extract of Meconopsis spinulosa is 10-11 mg / ml, the ethanol extract of Terminalia chebula is 1-2 mg / ml, and the ethanol extract of Crocus sativus is 48-49 mg / ml; or, the ethanol extract of Meconopsis spinulosa is 10-11 mg / ml, the ethanol extract of Terminalia chebula is 6-7 mg / ml, and the ethanol extract of Crocus sativus is 48-49 mg / ml.

5. The pharmaceutical composition according to claim 1, characterized in that The ethanol extract is an ethanol extract with a concentration of more than 95% V / V; further, the ethanol extract is a 95% V / V ethanol or anhydrous ethanol extract.

6. The method for preparing the pharmaceutical composition according to claim 1, characterized in that: It includes the following: (1) extracting each raw material with ethanol of corresponding concentration, concentrating and drying the extract to obtain each ethanol extract; (2) Mix the ethanol extracts according to the aforementioned weight portions to obtain the product.

7. Use of the pharmaceutical composition according to any one of claims 1 to 5 in the preparation of drugs for antibacterial or therapeutic bacterial infections.

8. The use according to claim 7, characterized in that The drug is a drug for resisting Klebsiella pneumoniae and / or Pseudomonas aeruginosa.

9. The use according to claim 7 or 8, characterized in that The drug is a drug against drug-resistant bacteria.

10. The use according to claim 9, characterized in that The drug-resistant bacteria are selected from: Klebsiella pneumoniae that is resistant to one or more of the following drugs: doxycycline, neomycin, tetracycline, oxytetracycline, ampicillin, penicillin, clindamycin, lincomycin, sulfamethoxazole, midecamycin, kanamycin; or / and, Pseudomonas aeruginosa resistant to one or more of the following drugs: cefotaxime, ceftriaxone, cefoperazone, polymyxin, mezlocillin, amoxicillin, tylosin, tetracycline, doxycycline, rifampicin, trimethoprim, tylosin.

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