CN115414487A - Application of cepharanthine and macrolide antibiotics in preparation of antibacterial products - Google Patents

Application of cepharanthine and macrolide antibiotics in preparation of antibacterial products Download PDF

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CN115414487A
CN115414487A CN202211089890.9A CN202211089890A CN115414487A CN 115414487 A CN115414487 A CN 115414487A CN 202211089890 A CN202211089890 A CN 202211089890A CN 115414487 A CN115414487 A CN 115414487A
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cepharanthine
group
erythromycin
streptococcus
combination
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CN115414487B (en
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李秀云
史文娜
谭冬梅
段雪霞
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Shandong Maternal And Child Health Hospital
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • 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
    • A61K31/4748—Quinolines; Isoquinolines forming part of bridged ring systems
    • 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
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7042—Compounds having saccharide radicals and heterocyclic rings
    • A61K31/7048—Compounds having saccharide radicals and heterocyclic rings having oxygen as a ring hetero atom, e.g. leucoglucosan, hesperidin, erythromycin, nystatin, digitoxin or digoxin
    • 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
    • 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
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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Abstract

The invention discloses application of cepharanthine and macrolide antibiotics in preparation of antibacterial products, and belongs to the technical field of medicines. The chessboard method is adopted to evaluate the in-vitro antibacterial activity of the combination of the cepharanthine and the erythromycin on the drug-resistant type B-group streptococcus, and researches show that the combination of the cepharanthine and the erythromycin shows a synergistic effect on the drug-resistant type B-group streptococcus, so that the dosage of the erythromycin can be remarkably reduced while the same antibacterial effect is achieved. The research result shows that the drug combination is expected to be applied to clinic and provides an idea for overcoming the current drug resistance situation of the clinical group B streptococcus.

Description

Application of cepharanthine and macrolide antibiotics in preparation of antibacterial products
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to application of cepharanthine and macrolide antibiotics in preparation of antibacterial products.
Background
The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.
Group B Streptococcus (GBS), also known as group B streptococcus or streptococcus agalactiae (s. Agalactiae), is one of the beta hemolytic aerobic gram-positive cocci. Group B streptococci can cause infection of pregnant women, premature fetal maldevelopment (low-body-weight infants), premature rupture of fetal membranes, late abortion and the like, and can also cause septicemia, pneumonia, meningitis and even death of newborn infants. At present, the prevention scheme for GBS at home and abroad mainly adopts antibiotic prevention, beta-lactam antibiotics are selected more, and macrolide antibiotics are commonly used for replacing patients with allergic constitution or slight infection. However, the use of macrolide antibiotics has increased dramatically worldwide, resulting in the emergence of a large number of resistant strains. The problem of drug resistance of GBS to macrolide antibiotics, especially erythromycin, is becoming more serious in our country, which brings great difficulty to clinical treatment and control of nosocomial infection.
Cepharanthin (Cepharanthin) is alkaloid separated from Stephania Japonica plant, is a leukocyte increasing drug, has effects of promoting peripheral leukocyte proliferation and certain function of proliferating platelet, and can be used for treating leukopenia due to chemotherapy and radiotherapy. In addition, cepharanthine has recently been reported as a potential drug for the treatment of viral infections. However, there has been no report on the antibacterial effect of the combination of cepharanthine and antibiotics.
Disclosure of Invention
In order to solve the defects of the prior art, the invention aims to provide the application of the cepharanthine and macrolide antibiotics in preparing antibacterial products. The chessboard method is adopted to evaluate the in-vitro antibacterial activity of the combination of the cepharanthine and the erythromycin on the drug-resistant type B streptococcus, and researches show that the combination of the cepharanthine and the erythromycin shows a synergistic effect on the drug-resistant type B streptococcus, so that the dosages of the cepharanthine and the erythromycin can be obviously reduced while the same bacteriostatic effect is achieved. The research result shows that the drug combination is expected to be applied to clinic and provides an idea for overcoming the current drug resistance situation of clinical group B streptococcus.
In order to realize the purpose, the technical scheme of the invention is as follows:
in a first aspect, there is provided the use of cepharanthine in combination with a macrolide antibiotic in the manufacture of an antibacterial product.
Cepharanthin (Cepharanthin) is alkaloid separated from Stephania Japonica plant, is a leukocyte increasing drug, has effects of promoting peripheral leukocyte proliferation and certain function of proliferating platelet, and can be used for treating leukopenia due to chemotherapy and radiotherapy. In addition, cepharanthine has also been recently reported as a potential drug for the treatment of viral infections, but there are no reports related to the treatment of group B streptococci. The invention discloses that combination of cepharanthine and macrolide antibiotics can improve the sensitivity of drug-resistant group B streptococcus to erythromycin, and has good curative effect on drug-resistant group B streptococcus.
The research of the invention shows that the combination of cepharanthine and erythromycin can significantly reduce the dosage of erythromycin.
Preferably, the macrolide antibiotics comprise erythromycin, erythromycin ethylsuccinate, azithromycin and roxithromycin.
More preferably, the macrolide antibiotic is erythromycin.
Preferably, the bacteria include, but are not limited to, group B streptococcus; further preferably, the group B streptococcus includes drug-resistant group B streptococcus.
In a second aspect, a pharmaceutical composition comprises cepharanthine and a macrolide antibiotic.
Preferably, in the pharmaceutical composition, the minimum inhibitory concentration of the cepharanthine is 8-32 mug/mL, and the minimum inhibitory concentration of the erythromycin is 0.0625-1 mug/mL.
In some embodiments of the present invention, the drug-resistant group B streptococcus is inhibited by a combination of cepharanthine and erythromycin, wherein the combination is at the respective concentrations of: the minimum inhibitory concentration of cepharanthine is 8-32 μ g/mL, and the minimum inhibitory concentration of erythromycin is 0.0625-1 μ g/mL. According to the experimental result, when the concentration of erythromycin is more than 0.0625-1 mug/mL and the concentration of cepharanthine is more than 8-32 mug/mL, the effect of synergistically resisting the drug-resistant type B streptococcus can be achieved when the bacteria infection caused by the drug-resistant type B streptococcus clinically is caused.
Preferably, the effective concentration ratio of the cepharanthine to the erythromycin is as follows: erythromycin =8-50:0.0625-10; the preferable mixture ratio is 8-32:0.0625-1.
In a third aspect, the use of the pharmaceutical composition of the second aspect in the preparation of a medicament for the prevention and treatment of diseases caused by group B streptococcus.
Preferably, the disease includes, but is not limited to, neonatal group B streptococcal disease, adult group B streptococcal disease.
Wherein said neonatal group B streptococcal disease includes both early onset and late onset types; early onset neonates have a time to onset within seven days of birth, including pulmonary infection, bacteremia, meningitis, and septicemia; late onset neonates, which are mostly within seven to three months of birth, include meningitis, endocarditis, arthritis, osteomyelitis, cellulitis, and septicemia.
The adult group B streptococcal disease includes maternal and maternal infections and infections in other adults; maternal infections including chorioamnionitis, endometritis, urinary infections, meningitis, endocarditis, and septicemia; other adult infections include pneumonia, meningitis, endocarditis, skin soft tissue infections, and sepsis.
In the medicine, the combination of cepharanthine and macrolide antibiotics is used as an active ingredient which accounts for 60-99% of the weight of the medicine, and the rest is a pharmaceutically acceptable carrier, diluent or solution or salt solution.
The present invention does not exclude combinations of one or more ingredients having the same or similar activity as the pharmaceutical composition, which combinations are likely to reduce the amount of the pharmaceutical composition in the composition, e.g. by 1-60% of the total weight, or, where the combination is particularly effective, to reduce the amount of the composition, e.g. by as little as less than 1% of the total weight, the skilled person being able to obtain the desired amount by routine experimentation as the case may be.
The medicaments of the invention may be in a variety of forms such as tablets, capsules, powders, syrups, solutions, suspensions and aerosols, and the like, and may be presented in suitable solid or liquid carriers or diluents and in suitable sterile devices for injection or instillation.
These pharmaceutical compositions are generally safe, non-toxic and biologically desirable, and thus, the pharmaceutically acceptable carriers or excipients described herein are non-toxic and safe. The pharmaceutically acceptable carriers and excipients of the present invention are generally well known to those skilled in the art or can be determined by those skilled in the art based on the circumstances.
The pharmaceutically acceptable excipients include, but are not limited to, excipients, carriers, and the like. One skilled in the art can select the appropriate excipient for various purposes. For example, depending on the nature of the drug or the mode of administration, commonly used excipients such as solvents, solubilizers, surfactants, preservatives, fillers, emulsifiers, binders, disintegrants, stabilizers, flavors, antioxidants, colorants, diluents, pH adjusters, pressure adjusters, and the like, or combinations of two or more of these, and the like.
The pharmaceutical composition may be administered in unit dosage form. The administration form may be a conventional form such as a liquid form such as an emulsion form, a colloid form, a true solution form, a microparticle form, a racemic form; such as other conventional machine types such as tablets, capsules, dripping pills, aerosols, pills, oral liquids, powders, injections, solutions, suspensions, emulsions, granules, clathrates, landfills, and the like. These dosage forms can be prepared by conventional methods of pharmaceutical manufacture, such as mixing, granulating, tableting, filling, dissolving or suspending dispersion, and the like.
The pharmaceutical composition of the present invention can be clinically used for mammals including human and animals, and can be administered by intravenous injection, oral, nasal, cutaneous, pulmonary, or gastrointestinal administration routes, etc. Regardless of the method of administration, the optimal dosage for an individual will depend on the particular treatment. Usually starting with a small dose and gradually increasing the dose until the most suitable dose is found.
In a fourth aspect, a method of treating group B streptococcal infection, the method comprising administering the pharmaceutical composition of the second aspect.
Compared with the prior art, the invention has the beneficial effects that:
cepharanthine is mainly used for leucopenia caused by various reasons such as chemotherapy, radiotherapy and the like in clinic, and at present, cepharanthine is not reported in the field of resisting group B streptococcus. The invention firstly provides the combination of cepharanthine and macrolide antibiotics for antibacterial treatment, and is expected to provide a thought for the development of antibacterial drugs.
The research on the combined application of cepharanthine and erythromycin shows that the combination of cepharanthine and erythromycin has obvious effect on drug-resistant group B streptococcus. Researches find that the cepharanthine can obviously improve the sensitivity of drug-resistant group B streptococcus to erythromycin, can greatly reduce the dosage of erythromycin if being expected to be applied to clinic, and has great clinical significance.
Detailed Description
It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an", and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
As the background suggests, the problem of GBS resistance to macrolide antibiotics, particularly erythromycin, is also growing, which presents great difficulties in clinical treatment and in the control of nosocomial infections. The invention establishes that the combination of cepharanthine and erythromycin has good in vitro antibacterial activity on group B streptococcus and shows synergistic effect through a chessboard method.
In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.
Example 1
Synergistic effect of combination of cepharanthine and erythromycin on drug-resistant group B streptococci
1 materials and methods
1.1 reagents and instruments
Erythromycin (HPLC ≥ 98%) was purchased from Dalian Meiren Biotechnology Ltd;
cepharanthine (HPLC ≥ 98%) was purchased from Yuanye biology, inc.;
THB medium and Mueller-Hinton agar medium were purchased from Haibo Biotechnology Ltd;
HKM-9802 Infrared inoculation sterilization circulator Guangdong Huanji Microbiol technologies, inc.;
DHP-81508 electric constant temperature incubator, jinan Ou Labock instruments, inc.;
VORTEX-GENIE2 VORTEX oscillator U.S. scientific industries;
STAR-VF-20 pure water apparatus Shanghai gravel pot Water treatment facilities, inc.;
pipette, us Eppendorf;
BSA124S electronic balance Sartorius, germany.
1.2 strains
3 strains of group B streptococcus (GroupBstreptococcus): GBS2, GBS3, GBS4;
2 method of experiment
2.1 chessboard method
2.1.1 working procedure
Referring to the CLSIM100-30 protocol, the drug stock solution was diluted to 2-fold concentration with THB liquid medium (final drug concentration range: erythromycin 0.0625-32. Mu.g/mL, cepharanthine 0.5-32. Mu.g/mL). The operation is carried out according to the following method: sequentially adding 50 mu L of diluted erythromycin from high concentration to low concentration into the 2 nd to 11 th columns of a 96-well plate; from low concentration to high concentration, 50 μ L of the diluted cepharanthine solution was added to the G-A line of a 96-well plate in order. 100 μ to each well except row 12 L bacterial suspension (10) 6 CFU/mL) and THB liquid culture based on wells with less than 200 μ L liquid. Wherein, the H1 wells are positive growth control wells, and the wells in row 12 are blank control wells. Culturing at 35 deg.C under constant temperature and humidity for 18h, and taking Minimum Inhibitory Concentration (MIC) in the lowest concentration well for macroscopic sterile growth.
2.1.2 method for evaluating effect of drug combination
The results were evaluated by FICI method. The method is a nonparametric method model developed on the basis of the Loeweaddivity (LA) theory. The formula is as follows:
FICI = MIC of a drug in combination/MIC of a drug in combination + MIC of B drug in combination/MIC of B drug in combination.
The experimental result is judged by adopting the standard that FICI is less than or equal to 0.5 for synergistic action, FICI is less than or equal to 0.5 for additive action, FICI is less than or equal to 1.0 for irrelevant action, and FICI is more than 2.0 for antagonistic action.
2.2 results of the experiment
2.2.1 results of the chessboard method experiment
TABLE 1 synergistic effect of the combination of cepharanthine and erythromycin on drug-resistant group B streptococci
Figure BDA0003836797530000081
The chessboard method experiment result shows that: when the cepharanthine and the erythromycin are used together for the drug-resistant strains of the group B streptococcus, the MIC of the cepharanthine and the erythromycin is reduced, the sensitivity of the strains to the erythromycin is enhanced, the strains become sensitive from drug resistance (MIC is more than or equal to 4), the MIC of the erythromycin after the combination is 0.0625-1 mug/mL, and the MIC of the cepharanthine is reduced to 8-32 mug/mL (Table 1). Based on MIC value, calculating FICI value after combination of two medicines, and determining as synergistic effect or additive effect.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (10)

1. The application of cepharanthine in combination with macrolide antibiotics in preparing antibacterial products is provided.
2. Use of cepharanthine in combination with a macrolide antibiotic according to claim 1, wherein said macrolide antibiotic comprises erythromycin, erythromycin ethylsuccinate, azithromycin, roxithromycin.
3. Use of cepharanthine in combination with a macrolide antibiotic according to claim 2, wherein said macrolide antibiotic is erythromycin, for the preparation of an antibacterial product.
4. Use of cepharanthine in combination with a macrolide antibiotic according to claim 1 for the manufacture of an antibacterial product, wherein said bacteria include but are not limited to group B streptococcus;
preferably, the group B streptococcus includes a drug-resistant group B streptococcus.
5. A pharmaceutical composition comprising cepharanthine and a macrolide antibiotic.
6. The pharmaceutical composition of claim 5, wherein the pharmaceutical composition has a minimum inhibitory concentration of cepharanthine of 8-32 μ g/mL and a minimum inhibitory concentration of erythromycin of 0.0625-1 μ g/mL.
7. The pharmaceutical composition of claim 6, wherein the effective concentration ratio of cepharanthine to erythromycin is cepharanthine: erythromycin =8-50:0.0625-10, preferably 8-32:0.0625-1.
8. Use of a pharmaceutical composition according to any one of claims 5 to 7 for the manufacture of a medicament for the prevention or treatment of a disease caused by group B streptococcus.
9. Use of a pharmaceutical composition according to claim 8 for the manufacture of a medicament for the treatment of a group B streptococcal disease, including but not limited to neonatal group B streptococcal disease, adult group B streptococcal disease;
wherein said neonatal group B streptococcal disease includes both early onset and late onset types; early onset neonates have a time to onset within seven days of birth, including pulmonary infection, bacteremia, meningitis, and septicemia; late onset neonatal morbidity, mostly seven to three months after birth, including meningitis, endocarditis, arthritis, osteomyelitis, cellulitis, and septicemia;
The adult group B streptococcal disease comprises maternal infection and infection of other adults; maternal infections including chorioamnionitis, endometritis, urinary infections, meningitis, endocarditis, and septicemia; other adult infections include pneumonia, meningitis, endocarditis, skin soft tissue infections, and sepsis.
10. A method of treating group B streptococcal infection, comprising administering a pharmaceutical composition according to any one of claims 5 to 7.
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Citations (4)

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Publication number Priority date Publication date Assignee Title
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CN106167493A (en) * 2016-07-06 2016-11-30 河北大学 The preparation method of novel cepharanthine and the application on medicine thereof
US9539305B1 (en) * 2014-03-14 2017-01-10 Fleurir Abx Llc Pristinamycin compositions, LpxC compositions, their improvements, and combinations thereof
CN110448554A (en) * 2019-09-12 2019-11-15 济南市儿童医院（山东大学齐鲁儿童医院） Application of D-penicillamine and fluconazole in preparation of antifungal product

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Publication number Priority date Publication date Assignee Title
US9539305B1 (en) * 2014-03-14 2017-01-10 Fleurir Abx Llc Pristinamycin compositions, LpxC compositions, their improvements, and combinations thereof
CN105457084A (en) * 2015-12-17 2016-04-06 张德信 Surgical suture with analgesic and antibacterial effect and preparation method thereof
CN106167493A (en) * 2016-07-06 2016-11-30 河北大学 The preparation method of novel cepharanthine and the application on medicine thereof
CN110448554A (en) * 2019-09-12 2019-11-15 济南市儿童医院（山东大学齐鲁儿童医院） Application of D-penicillamine and fluconazole in preparation of antifungal product

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Title
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