CN112294789B - Compound preparation for preventing and treating respiratory tract infection and application thereof - Google Patents

Compound preparation for preventing and treating respiratory tract infection and application thereof Download PDF

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CN112294789B
CN112294789B CN202011101034.1A CN202011101034A CN112294789B CN 112294789 B CN112294789 B CN 112294789B CN 202011101034 A CN202011101034 A CN 202011101034A CN 112294789 B CN112294789 B CN 112294789B
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chlorhexidine
respiratory tract
compound preparation
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CN112294789A (en
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郑涛
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    • 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
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid, pantothenic acid
    • A61K31/198—Alpha-aminoacids, e.g. alanine, edetic acids [EDTA]
    • 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
    • A61K31/13—Amines
    • A61K31/155—Amidines (), e.g. guanidine (H2N—C(=NH)—NH2), isourea (N=C(OH)—NH2), isothiourea (—N=C(SH)—NH2)
    • 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
    • A61P11/00—Drugs for disorders of the respiratory system
    • A61P11/02—Nasal agents, e.g. decongestants
    • 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
    • A61P11/04—Drugs for disorders of the respiratory system for throat disorders
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
    • A61P15/02—Drugs for genital or sexual disorders; Contraceptives for disorders of the vagina
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • 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
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • 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

Abstract

The invention provides a compound preparation for preventing and treating respiratory tract infection, which comprises the following raw materials: a. one or more of amino acid and analogues thereof; b. one or more of chlorhexidine and analogs thereof; wherein the mass ratio of the raw material a to the raw material b is (0.01-100): (0.01-100). The compound preparation can effectively prevent respiratory tract infection and has the advantages of strong antibacterial/antiviral property, wide antibacterial/antiviral spectrum, low toxic and side effects and the like.

Description

Compound preparation for preventing and treating respiratory tract infection and application thereof
Technical Field
The invention relates to a medicament for preventing and treating respiratory tract infection, in particular to a compound preparation for preventing and treating respiratory tract infection and application thereof, belonging to the field of medicines.
Background
Respiratory tract infection mainly refers to inflammation caused by infection of upper respiratory tract (such as nose, ear, sinus and throat) and/or lower respiratory tract (such as trachea, bronchus and lung) by virus/bacteria/fungi, for example, acute upper respiratory tract infection is a generic name of acute inflammation from nasal cavity to larynx, which is also one of more common infectious diseases, and researches show that common microorganisms/viruses causing respiratory tract infection mainly include pneumococci, staphylococcus aureus, escherichia coli, streptococcus b, influenza a virus, influenza b virus and the like. For example, around 90% of influenza is caused by influenza virus, bacterial infection occurs secondary to viral infection, influenza occurs throughout the year (most common in winter and spring), and can occur in people of any age group, seriously affecting human health and life.
Common respiratory tract infection diseases mainly comprise cold, acute/chronic tracheitis/bronchitis/pneumonia, acute/chronic rhinitis/nasosinusitis, tonsillitis and the like, and the application of antibacterial/antiviral drugs to an infected organism is an effective means for preventing and treating the respiratory tract infection diseases, however, most of the conventional drugs have the defects of large toxic and side effects, easy generation of drug resistance in the organism and the like, narrow antibacterial/antiviral spectrum, single effect and limited application range, so that the development of a novel pharmaceutical preparation for improving the prevention and treatment effect, expanding the application range, reducing the toxic and side effects, drug resistance and the like is still an important subject and challenge for technicians in the field.
In addition, research and development of novel pharmaceutical preparations provide more drug choices, and the development of new viruses and diseases caused by the new viruses is urgent, for example, new coronavirus pneumonia (COVID-19) occurring in 2019 brings great training and challenge to human beings, COVID-19 is acute infectious pneumonia of respiratory system caused by infection of new coronavirus, the new coronavirus is mainly spread through respiratory droplets and direct contact, respiratory tract and nasal mucosa are the first damaged parts, a large amount of viruses are replicated and shed in respiratory tract epithelial cells in early infection, the viruses can be detected in the epithelial cells in about 96 hours after infection, the main clinical symptoms of patients include fever, dry cough, nasal mucus, sore throat, olfactory decline, hypodynamia and the like, as the disease progresses, a large amount of mucus is generated in large and small bronchi and mucus plugs are formed, a cytokine storm exists in a heavy and critical patient, an immune system is seriously damaged, and secondary pulmonary fungus, bacteria or fungus and bacteria mixed infection is one of important causes of death of the critical patient (see a novel diagnosis and treatment scheme (trial seventh edition) for coronavirus pneumonia), unfortunately, no specific medicine for treating respiratory system infection caused by the novel coronavirus exists at present. With the development of human society, especially during and after the covi-19 epidemic situation, the human health defense enters the normalization, and the reasonable administration, the selective administration, the effective administration and the preventive administration will be concerned more and more widely, and the provision of more and more effective pharmaceutical preparations for preventing and treating common diseases such as respiratory tract infection and the like is more important.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a compound preparation for preventing and treating respiratory tract infection, which has the advantages of strong antibacterial/antiviral property, wide antibacterial/antiviral spectrum, low toxic and side effects, difficulty in causing drug resistance of organisms and the like, can effectively prevent and treat respiratory tract infection, and overcomes the defects in the prior art.
In one aspect of the invention, a compound preparation for preventing and treating respiratory tract infection is provided, which comprises the following raw materials: a. one or more of amino acid and analogues thereof; b. one or more of chlorhexidine and analogs thereof; wherein the mass ratio of the raw material a to the raw material b is (0.01-100): (0.01-100).
The compound preparation provided by the invention has the advantages of strong antibacterial/antiviral property, wide antibacterial/antiviral spectrum, low toxic and side effects, difficulty in generating drug resistance in organisms and the like due to the synergistic cooperation of the components, can effectively prevent and treat respiratory tract infection, and particularly can prevent and treat upper respiratory tract infection diseases such as cold, rhinitis, nasosinusitis, tonsillitis and the like.
The inventor considers through research and analysis that the amino acid component (the raw material a) can participate in the synthesis of body protein, stimulate the release of growth hormone, have a nutritional effect on local inflammation, expand local blood vessels, improve the local blood supply condition of inflammation, promote the recovery of inflammation, be used as an immunopotentiator, increase the immune function of T cells, promote the immune system to secrete natural killer cells, phagocytes, interleukin (Interleukin-1) and other endogenous substances, and be beneficial to improving the anti-infection capability of the body, and the amino acid component can achieve the broad-spectrum antibacterial/viral and bactericidal/viral effects and effectively prevent and treat diseases such as respiratory tract infection by being cooperatively matched with the chlorhexidine component (the raw material b). In addition, the results of years of clinical use show that arginine and chlorhexidine do not generate drug resistance, and amino acid (which is more commonly used as a pharmaceutic adjuvant) and chlorhexidine (such as chlorhexidine acetate disinfectant preservatives and the like) generate few adverse reactions.
Further, in the compound preparation, the mass ratio of the raw material a to the raw material b can be (0.01-50): (0.01-50), for example, (0.1-50): (0.1-50) or (1-50): (1-50) or (10-50): (10-50) or (12-50): (12-50).
The compound preparation can be in the dosage forms of medical conventional liquid agents (such as solution, emulsion, suspension and the like), gels, solid agents and the like, such as spray, hand sanitizer, disinfectant, powder mist and the like, and can be prepared by adopting the conventional method in the field and corresponding dosage form auxiliary materials (or excipients).
In an embodiment of the present invention, the raw materials of the compound preparation include, by 100 parts by weight, 0.01 to 10 parts of raw material a, 0.01 to 10 parts of raw material b, and the balance of auxiliary materials, and the compound preparation may be generally a solid dosage form such as powder. Wherein, the raw material a can be 0.1 to 10 parts or 1 to 10 parts, and the raw material b can be 0.1 to 10 parts or 1 to 10 parts.
In a preferred embodiment of the present invention, the compound reagent may be a liquid preparation or a gel preparation, wherein the content of the raw material a is 0.01 to 100mg/ml, further 0.1 to 50mg/ml, or 1 to 50mg/ml, or 10 to 50mg/ml, or 12 to 50mg/ml, the content of the raw material b is 0.01 to 100mg/ml, further 0.1 to 50mg/ml, or 1 to 50mg/ml, or 10 to 50mg/ml, or 12 to 50mg/ml, and the balance may be an auxiliary material.
Specifically, the compound preparation can be a nasal drop, specifically a gel nasal drop (or a gel nasal drop), is used for local administration on the upper respiratory tract nasal mucosa, a pathogenic bacteria prevention and treatment wall (also called build Di, the gel nose drop can be called build Di gel nose drop) is locally constructed on the nasal mucosa of the upper respiratory tract, on one hand, the pathogenic bacteria can be prevented from entering the body, on the other hand, the proper pH environment can be locally constructed, the effective components of the medicine are fully released, the medicine with higher concentration is adhered to the local part, the contact time of the medicine and the nasal mucosa of the upper respiratory tract is increased, and the gel nasal drop is locally applied to form a film adhered to the surface of inflammation, the medicine can deeply enter a subcutaneous tissue layer in a short time to quickly exert the medicine effect, thereby achieving the effects of improving the treatment effect, expanding the antibacterial/antiviral spectrum, reducing the morbidity and the severe transformation rate and the like.
According to further research of the present invention, the amino acid may be at least one selected from arginine, leucine, glutamic acid, ornithine and lysine (lysine), and the material a may be one or a mixture of several of the amino acids and/or analogues thereof, so as to improve characteristics of the compound preparation, such as antibacterial/antiviral property, antibacterial/antiviral spectrum, and the like. Specifically, in one embodiment of the present invention, the amino acid is arginine, which can stimulate the release of growth hormone, promote the absorption of inflammation, enhance the immune function, and prevent viral infection, thereby improving the local anti-infection ability.
In addition, the chlorhexidine can be chlorhexidine acetate, which is beneficial for multi-target attack of compound preparation, expansion of antibacterial/antiviral spectrum, and enhancement of antibacterial/antiviral effect.
In the present invention, the amino acid analog may be a substance having a structure similar to that of an amino acid in the conventional art, for example, an amino acid derivative such as a salt of an amino acid, and similarly, the chlorhexidine analog may be a substance having a structure similar to that of chlorhexidine in the conventional art, for example, a chlorhexidine derivative such as a salt of chlorhexidine. In one embodiment of the invention, amino acid analogs include salts of amino acids; and/or the chlorhexidine analogue comprises a salt formed by chlorhexidine, and is beneficial to the prevention and treatment effect of the compound preparation on respiratory tract infection. It is understood that the amino acid and the analogues thereof, the chlorhexidine and the analogues thereof and other raw materials are medicinal raw materials, and the salt formed by the amino acid and the salt formed by the chlorhexidine are medicinal salts.
Further, the chlorhexidine forming salt may specifically include a chlorhexidine organic acid salt, such as may include chlorhexidine digluconate or chlorhexidine diacetate or a mixture of both.
On the other hand, the invention also provides application of the compound preparation in preparing a medicament for preventing and treating respiratory tract infection diseases.
Specifically, the compound preparation can be used as a medicine for preventing and treating diseases such as respiratory tract infection and the like, or used as a raw material medicine for preparing the medicine for preventing and treating the diseases, and particularly has obvious prevention and treatment effects on the upper respiratory tract infection diseases.
The respiratory tract infection diseases include diseases caused by microbial and/or viral infection, wherein the microbial can be bacteria and/or fungi, such as common gram-positive bacteria and gram-negative bacteria. According to the research of the present invention, the above microorganisms and/or viruses may include one or more of pneumococci, staphylococcus aureus, escherichia coli, streptococcus b, novel coronaviruses, and influenza a and b viruses. Research shows that the compound preparation has stronger inhibiting and killing effects on the microbial bacteria and/or viruses, has stronger sterilizing and disinfecting effects compared with the conventional medicaments such as Xinjieer, ribavirin and the like, and has the advantages of small toxic and side effects, wide antibacterial/antiviral spectrum, difficulty in causing the organisms to generate drug resistance and the like.
In one embodiment of the present invention, the compound preparation is a bundi gel nasal drop, and the in vivo prevention and treatment effects thereof are as follows through pharmacodynamic tests: the build Di gel nose drops can obviously prolong the survival days of mice infected by microorganisms such as staphylococcus aureus, pneumococcus and the like, and improve the number of survival animals, so that the build Di gel nose drops have good protection effect on the mice infected by microorganisms such as staphylococcus aureus, pneumococcus and the like; its in vitro antibacterial activity is as follows: the build Di gel nose drop has obvious antibacterial and bactericidal effects on Diplococcus pneumoniae, Staphylococcus aureus, Escherichia coli, Streptococcus B and other microorganisms, and also has strong inhibiting and killing effects on influenza A virus and influenza B virus.
The above respiratory tract infection diseases may include one or more of common cold, rhinitis, sinusitis, tonsillitis, tracheitis, bronchitis, and pneumonia, such as acute/chronic rhinitis, acute/chronic sinusitis, acute/chronic tracheitis, and acute/chronic bronchitis. In addition, through further experimental study, the compound preparation also has strong inhibiting and killing effects on the novel coronavirus, so that the compound preparation is expected to be used for preventing and treating COVID-19 pneumonia caused by the virus.
In another aspect of the invention, the invention also provides an application of the compound preparation in preparing a medicament for preventing and treating vaginitis and/or cervicitis.
The inventor researches and discovers that the compound preparation not only has good prevention and treatment effects on respiratory tract infection diseases, but also is suitable for preventing and treating vaginitis and/or cervicitis, and particularly has more remarkable prevention and treatment effects on external negative vaginitis and/or cervicitis. The compound preparation can be directly used as a medicine for preventing and treating vaginitis and/or cervicitis, or can be used as a raw material medicine for preparing the medicine for preventing and treating the diseases, and as mentioned above, the compound preparation has the advantages of strong antibacterial/antiviral property, wide antibacterial/antiviral spectrum, low toxic and side effects, difficulty in causing an organism to generate drug resistance and the like, so that the compound preparation has greater practical significance.
The implementation of the invention has at least the following beneficial effects:
the compound preparation is prepared by compounding the amino acids and the chlorhexidine, belongs to a non-hormone non-antibiotic preparation, reduces the risks of drug resistance and side effects such as cortical hyperfunction and the like, has stronger antibacterial/antiviral property and wider antibacterial/antiviral spectrum, has wide application range, stronger lipophilicity, is beneficial to the absorption of organisms, takes effect quickly, can effectively prevent and treat diseases such as respiratory tract infection and the like, and is beneficial to practical application and popularization.
Detailed Description
The present invention will be described in more detail with reference to examples. It is to be understood that the practice of the invention is not limited to the following examples, and that any variations and/or modifications may be made thereto without departing from the scope of the invention.
Example 1
The compound preparation provided by the embodiment is a build di gel nasal drop, which is composed of the following raw materials: 1mg/ml of arginine, 1mg/ml of chlorhexidine and the balance of auxiliary materials. The compound preparation is prepared by adopting conventional gel nasal drops auxiliary materials and conventional methods in the field, and is not described again.
Example 2
The antibacterial/antiviral properties of the tyndallia gel nasal drops were examined by the following tests, which were conducted using the following reagents/drugs, mouse, microorganism, virus, and cell sources, unless otherwise specified:
chlorhexidine, manufactured by Jiutai pharmaceuticals, Inc. of Jinzhou;
arginine, manufactured by Shandong Tangzheng Biotech, Inc.;
ciprofloxacin, manufactured by south new drug manufacturing, guangzhou, inc;
ribavirin: manufactured by north china pharmaceutical limited;
kunming (KM) mice purchased from the animal center of the military medical science research institute of the military scientific institute, all mice can be freely eaten and have sufficient fresh drinking water, the feeding environment of the mice meets the environmental facility standard of the animal grade of SPF experiments, and the mice are approved by the animal ethics committee of the military medical science research institute of the military scientific institute;
standard strains such as staphylococcus aureus, escherichia coli, bacillus subtilis and the like are provided by the identification of microorganisms of the Ministry of health;
the clinical separated strains of staphylococcus aureus, escherichia coli, pneumococcus and streptococcus type B are provided by the separation of a first hospital microbe room affiliated to the medical college of Zhejiang university;
influenza A virus A/PR8/34(H1N1), influenza B virus B/Jingfang 98-76, purchased from the institute of preventive medicine sciences virus of China;
MDCK cells, purchased from the biomedical research institute of military medical science institute.
In the following experiments, mice were used, group housed, water ad libitum, housed in the environment prior to dosing: room temperature (about 22-25 ℃), humidity 40-60%; the statistical methods of statistical significance involved in the following experiments mainly include t-test (measurement data) and chi-square test (counting data), which are conventional methods in the art and are not described in detail.
Test 1 pharmacodynamic test (in vitro antibacterial Activity test)
1.1 preparation of bacterial liquid
Respectively inoculating standard strains of staphylococcus aureus, escherichia coli and bacillus subtilis bud bacillus and clinical isolated strains of staphylococcus aureus, escherichia coli, pneumococcus and streptococcus B in a meat broth culture medium, and culturing at 37 ℃ for 18 hours; taking nutrient broth culture of each strain cultured for 18 hr, and culturing the nutrient broth according to the ratio of 10-3Diluting to obtain 8 bacterial liquids.
1.2 Experimental procedure of two-fold dilution method (test tube method)
1) Examples of the experiments
(1) Taking 11 sterilized test tubes, adding 1.6ml of nutrient broth liquid culture medium into the 1 st test tube, adding 1ml of nutrient broth liquid culture medium into each of the other test tubes, adding 0.4ml of the bundy gel nasal drops of the example 1 into the 1 st test tube, uniformly mixing, adding 1ml of the mixture in the 1 st test tube into the 2 nd test tube, uniformly mixing, adding 1ml of the mixture in the 2 nd test tube into the 3 rd test tube … …, sequentially diluting until 1ml of the mixture in the 9 th test tube is added into the 10 th test tube, uniformly mixing, sucking 1ml of the mixture out of the 10 th test tube, and discarding; no drug solution (i.e. gel nasal drops or diluent thereof) is added into the 11 th test tube as a control; then, 0.1ml of the bacterial suspension was added to each test tube, and cultured at 37 ℃ for 20 hours, and the Minimum Inhibitory Concentration (MIC) was observed.
The 8 bacterial liquids prepared in 1.1 were each tested according to the procedure of the step (1).
2) Comparative test
Comparative example 1: adopting chlorhexidine acetate single drug to replace build digel nasal drop to carry out the experimental process of 1.2, and taking the positive control;
comparative example 2: replacing build gel nose drops with arginine single drug to carry out the experimental process of 1.2, and taking the experimental process as a positive control;
comparative example 3: the experimental procedure 1.2 was carried out using ciprofloxacin instead of tsukuidi gel nasal drops as a positive control.
1.4 test results
The test corresponding to the 1 st to 10 th test tubes in the experimental example 1 shows good inhibition effect on the standard strain and the clinical isolated strain, and is superior to the comparative examples 1 to 3, especially the following cases are more remarkable:
in the test tube, when the concentration of the bundi gel nasal drops is more than 1.25ug/ml (the concentration refers to that the sum of the mass of arginine and chlorhexidine acetate in 1ml of test tube mixture is 1.25ug) (the test corresponding to the 1 st to 9 th test tubes), the bundi gel nasal drops have obvious bacteriostatic action on staphylococcus aureus, streptococcus B, bacillus subtilis and the like, and are superior to the comparative examples 1 to 3;
when the concentration of the bundi gel nasal drops is more than 2.5ug/ml (the test corresponding to the 1 st to 8 th test tubes), the bundi gel nasal drops have obvious bacteriostatic action on pneumococci and are superior to the comparative examples 1 to 3;
when the concentration of the build gel nasal drops is more than 5.0ug/ml (test corresponding to 1 st to 7 th test tubes), the build gel nasal drops have obvious bacteriostatic action on escherichia coli and are superior to comparative examples 1 to 3.
Table 1 below shows MICs corresponding to the 2 nd test tubes (n ═ 2) of the experimental examples and comparative examples 1 to 350As a result, the antibacterial effect of the build-up gel nose drops is further more visually shown.
Table 1 results of in vitro antibacterial activity test (n ═ 2) of experimental examples and comparative examples 1 to 3
Figure BDA0002725383070000081
Experiment 2 protection experiment of build digel nose drops on microorganism bacterium infected mouse
2.1 preparation of bacterial liquid
Inoculating clinically isolated staphylococcus aureus into a nutrient broth culture medium, and culturing for 18h in an incubator at 37 ℃; taking out, streaking on blood plate (10% sheep red blood cell nutrition agar plate), and culturing in 37 deg.C incubator for 20 hr; taking out, eluting bacterial colony with 6ml of 5% gastrodin (pH7.2) to obtain bacterial liquid A0, washing a blood plate with 4ml of physiological saline solution, adding the liquid after washing the blood plate into the bacterial liquid A0, and blowing the bacterial liquid A0 to disperse the bacterial colony; taking 4 parts of bacterial liquid A0, and preparing bacterial liquids A1, A2, A3 and A4 with different concentrations by using physiological saline respectively, wherein the specific steps are as follows:
a1 is prepared from 0.33ml bacterial liquid A0 and 1ml normal saline (the concentration of A1 is recorded as 0.33 ml/ml);
a2 is prepared from 0.20ml bacterial liquid A0 and 1ml normal saline (the concentration of A2 is recorded as 0.20 ml/ml);
a3 is prepared from 0.11ml bacterial liquid A0 and 1ml normal saline (the concentration of A3 is recorded as 0.11 ml/ml);
a4 was prepared from 0.06ml of bacterial solution A0 and 1ml of physiological saline (the concentration of A4 was 0.06 ml/ml).
2.2 screening of appropriate bacterial liquid concentration
Taking 40 KM mice (the weight is about 18-22g, and the weight is half each male and female), averagely dividing the KM mice into 4 groups (half each female and male in each group), and respectively marking the groups as a group 1a, a group 2a, a group 3a and a group 4 a; injecting a bacterium solution A1 into the mice of the group 1a, injecting a bacterium solution A2 into the mice of the group 2a, and injecting a bacterium solution A3 into the mice of the group 3 a; mice of group 4a were injected with bacterial suspension a4, and each mouse was injected with 0.5ml of bacterial suspension, and the number of deaths of infected mice was observed.
The results show that the mortality rates of the mice infected by the groups 1a to 4a are respectively 100%, 90% and 40%, and according to the results, the bacteria solution (A3) with the concentration of 0.11ml/ml is selected for subsequent tests to test the protective effect of the tyndado gel nasal drops on the mice infected by the staphylococcus aureus.
2.3 dosing test procedure
Taking KM mice (with the weight of 18-22g), averagely dividing into 8 groups (a control group 1a-5a and a test group 1a-3a), wherein each group comprises 20 mice (each half of male and female), respectively performing intragastric administration on each group of mice, and continuously administering for 5 days, wherein the administration volume is 10ml/kg and 1 time/day; wherein, 1h after 3 days of administration, except for a blank control group (namely a control group 1a), all the other groups are injected with golden yellow staphylococcus bacteria liquid (A3) in an intraperitoneal way, the injection amount is 0.5ml of bacteria liquid per mouse, then the administration is continued for 2 days, and the death condition of each group of animals within 7 days is observed. Wherein the medicines used in each group are as follows:
control group 1 a: an equivalent amount of physiological saline (administration volume: 10 ml/kg);
control group 2 a: an equivalent amount of physiological saline (administration volume: 10 ml/kg);
control group 3 a: ciprofloxacin, 0.5g/10 ml/kg;
control group 4 a: chlorhexidine acetate, 0.5g/10 ml/kg;
control group 5 a: arginine, 0.5g/10 ml/kg;
test group 1 a: 0.031g/10ml/kg of bundi gel nasal drop;
test group 2 a: 0.125g/10ml/kg of build Di gel nose drops;
test group 3 a: 0.5g/10ml/kg of build Di gel nose drops.
In the above control group, "0.5 g/10 ml/kg" means that the administration volume is 10ml/kg, and the content of the effective component in each 10ml of the preparation is 0.5g, and unless otherwise specified, the following 0.5g/10ml/kg means the same.
In the test group, 0.031g/10ml/kg represents a dosage capacity of 10ml/kg, and each 10ml of the preparation contains 0.031g arginine and 0.031g chlorhexidine acetate; 0.125g/10ml/kg represents a 10ml/kg administration volume, containing per 10ml of the medicament 0.125g arginine and 0.125g chlorhexidine acetate; 0.5g/10ml/kg indicates a 10ml/kg administration volume, containing 0.5g arginine and 0.5g chlorhexidine acetate per 10ml of the medicament. Unless otherwise specified, the following terms 0.031g/10ml/kg, 0.125g/10ml/kg and 0.5g/10ml/kg mean the above meanings, respectively.
2.4 results of the experiment
The average survival days of the mice in each group are shown in Table 2, and it can be seen that the survival numbers and the average survival days of the test groups 1a-3a are better than those of the control groups 1a-5a, especially the test groups 2a and 3a have more significant effects, and the difference between the survival numbers and the survival days of the test groups 2a-3a has larger statistical significance (P is less than 0.05) compared with the control groups 3a-5 a.
TABLE 2 protective Effect of groups on Staphylococcus aureus infected mice
Group of Animal number (only) Dosage (g/kg) Death number (only) Survival number (only) Number of days of survivald
Control group 1a 20 - 0 20 7.00±0.00
Control group 2a 20 - 18 2※※ 3.07±1.07※※
Control group 3a 20 0.5 11 9☆☆ 4.55±1.13☆☆
Control group 4a 20 0.5 9 11 4.89±0.78
Control group 5a 20 0.5 10 10 4.40±1.17
Test group 1a 20 0.031+0.031 5 15 5.20±0.84
Test group 2a 20 0.125+0.125 3 17**◇○ 6.33±0.58*◇○
Test group 3a 20 0.5+0.5 2 18**◇○○ 6.50±0.71*◇○
The corresponding color is that P is less than 0.01 compared with the contrast group 1 a; it means that P < 0.01 compared to control 2 a; p < 0.05 compared to control 3 a; p < 0.05 compared with the reference group 4 a; o represents P < 0.05 as compared with the control group 5 a; d represents data on days of survival as mean ± standard deviation.
Test 3 protective Effect on Diplococcus pneumoniae infected mice
3.1 preparation of bacterial liquid
Inoculating the clinically isolated pneumococcus into a culture medium of a tube-embedded broth, and culturing in an incubator at 37 ℃ for 18 h; taking out, streaking on a blood plate (1% sheep red blood cell nutrient agar plate), and culturing in an incubator at 37 deg.C for 24 h; taking out, eluting bacterial colonies by 6ml of 5% gastrodin (pH7.2) to obtain bacterial liquid B0, washing a blood plate by 4ml of physiological saline solution, adding the liquid after washing the blood plate into the bacterial liquid B0, and blowing the bacterial liquid B0 to disperse the bacterial colonies; taking 4 parts of bacterial liquid B0, and preparing bacterial liquids B1, B2, B3 and B4 with different concentrations by using physiological saline respectively, wherein the specific steps are as follows:
b1 is prepared from 1ml bacterial liquid B0 and 1ml normal saline (the concentration of B1 is recorded as 1 ml/ml);
b2 is prepared from 0.50ml bacterial liquid B0 and 1ml normal saline (the concentration of B2 is recorded as 0.5 ml/ml);
b3 is prepared from 0.33ml bacterial liquid B0 and 1ml normal saline (the concentration of B3 is recorded as 0.33 ml/ml);
b4 was prepared from 0.25ml of bacterial liquid B0 and 1ml of physiological saline (the concentration of B4 was 0.25 ml/ml).
3.2 screening of appropriate bacterial liquid concentration
Taking 40 KM mice (the weight is about 18-22g, and the weight is half each male and female), averagely dividing the KM mice into 4 groups (half each female and male in each group), and respectively marking the groups as a group 1b, a group 2b, a group 3b and a group 4 b; injecting a bacterium solution B1 into the mice of the group 1B, injecting a bacterium solution B2 into the mice of the group 2B, and injecting a bacterium solution B3 into the mice of the group 3B; mice in group 4B were injected with bacterial suspension B4, 0.5ml of bacterial suspension per mouse, and the number of deaths of infected mice was observed.
The results show that the mortality rates of the mice infected by the groups 1B-4B are respectively 100%, 60% and 30%, according to the results, the subsequent test can be performed by using 0.45ml/ml bacterial solution (prepared by 0.45ml bacterial solution B0 and 1ml normal saline, and the mortality rate of the mice infected by the bacterial solution with the concentration of 0.45ml/ml is estimated to be between 80% and 90%), and the protective effect of the tyndallic gel nasal drop on the mice infected by the pneumococci is tested.
3.3 dosing test procedure
Taking 40 KM mice (with the weight of 18-22g), averagely dividing the KM mice into 8 groups (a control group 1b-5b and a test group 1b-3b), wherein each group comprises 20 mice (each half of male and female), respectively performing intragastric administration on each group of mice, and continuously administering the KM mice for 5 days, wherein the administration volume is 10ml/kg and 1 time/day; wherein, 1h after 3 days of administration, except for a blank control group (namely a control group 1b), all the other groups are injected with the bacterial liquid with the concentration of 0.45ml/ml in an intraperitoneal way, the injection amount is 0.5ml of the bacterial liquid/mouse, then the administration is continued for 2 days, and the death condition of each group of animals within 7 days is observed. Wherein the medicines used in each group are as follows:
control group 1 b: an equivalent amount of physiological saline (administration volume: 10 ml/kg);
control group 2 b: an equivalent amount of physiological saline (administration volume: 10 ml/kg);
control group 3 b: ciprofloxacin, the administration dose is 0.5g/10 mg/kg;
control group 4 b: chlorhexidine acetate, administration dosage 0.5g/10 ml/kg;
control group 5 b: arginine, the dosage of administration is 0.5g/10 ml/kg;
test group 1 b: the build Di gel nose drop has the administration dosage (0.031g arginine +0.031g chlorhexidine acetate) of 0.031g/10 ml/kg;
test group 2 b: build Di gel nose drop, the administration dosage (0.125g arginine +0.125g chlorhexidine acetate) 0.125g/10 ml/kg;
test group 3 b: the build Di gel nose drops, the administration dosage (0.5g arginine +0.5g chlorhexidine acetate) 0.5g/10 ml/kg.
3.4 test results
As shown in Table 3, the survival days of the mice in each group are better than those in the control groups 1b-5b, especially the test groups 2b and 3b have more significant effect, and the difference between the survival days and the survival days of the test groups 2b-3b has larger statistical significance (P is less than 0.05) compared with the control groups 3b-5 b.
TABLE 3 protective Effect of groups on Diplococcus pneumoniae infected mice
Group of Animal number (only) Dosage (g/kg) Death number (only) Survival number (only) Number of days of survivald
Normal control group 20 - 0 20 7.000±0.000
Model set 20 - 17 3※※ 3.23±1.30※※
Ciprofloxacin 20 0.5 10 10☆ 4.40±1.07☆☆
Chlorhexidine group 20 0.5 8 12 4.75±0.71
Arginine group 20 0.5 9 11 4.22±1.20
Build di 1 group 20 0.031+0.031 7 13 5.14±0.69
Build di 2 group 20 0.125+0.125 4 16* 6.00±0.82*◇○
Build di 3 groups 20 0.5+0.5 2 18**◇○ 6.50±0.71*◇○
The corresponding color is that P is less than 0.01 compared with the contrast group 1 b; it means that P < 0.01 compared to control 2 b; comparison with control 3b indicates P < 0.05; let it mean that P < 0.05 is compared with the control 4 b; o represents P < 0.05 as compared with the control group 5 b; d represents data on days of survival as mean ± standard deviation.
Experiment 4 antiviral experiment (in vitro) of build Di gel nose drops
Test groups: in MDCK plates grown as monolayers, each well (see well numbers 1-5 in Table 2) was infected with 100TCID for influenza A or B virus50Adsorbing the virus solution for 1h, and removing the virus solution to obtain cell sap; adding bundi gel nasal drops into the cell sap at 37 deg.C and 5% CO2Culturing in an incubator, and observing cytopathic effect.
Six different drug concentration tests were carried out according to the above procedure, with drug concentrations of 80. mu.g/ml, 40. mu.g/ml, 20. mu.g/ml, 10. mu.g/ml, 5. mu.g/ml, 2.5. mu.g/ml (see Takedi gel nose drops test group (. mu.g/ml) in Table 4); the drug concentration refers to the mass of the build gel nasal drops added per milliliter of cell sap (for example, 80 mug/ml refers to 80 mug of the build gel nasal drops added into 1ml of cell sap); wherein, in the build Di gel nasal drops, the mass content of arginine and chlorhexidine acetate is 100 mug/ml respectively.
According to the test process of the test group, the following control groups are designed at the same time:
virus control group: the control group is different from the test group only in that no drug is added after the cell fluid is infected with the virus;
arginine control group: the control group and the test group are only different in that the build digel nose drops are replaced by arginine single drug, and the concentration of the drug is 100 mug/ml (the concentration refers to that 100 mug arginine is added into per ml of cell sap);
chlorhexidine control group: the control group is different from the test group only in that the build gel nasal drops are replaced by chlorhexidine acetate single drug, and the drug concentration is 100 mug/ml (the concentration refers to that 100 mug of chlorhexidine acetate is added in each ml of cell sap);
ribavirin control group: the control group is different from the test group only in that ribavirin is adopted to replace the zadi gel nasal drops, and the drug concentration is 10mg/ml (the concentration refers to that 10mg of ribavirin drug is added into per ml of cell sap);
cell control group: the control group differed from the test group only in that it was not infected with virus.
The results are shown in tables 4 and 5: as can be seen from Table 4, when the concentration of the bundy gel nasal drops is more than 10 mug/ml, the toxic effect of influenza A virus on MDCK cells can be effectively inhibited, and the effect is superior to that of an arginine single-drug and chlorhexidine single-drug control group; as can be seen from Table 5, when the concentration of the bundy gel nasal drops is more than 20 mug/ml, the toxic effect of the influenza B virus on MDCK cells can be inhibited, and the effect is better than that of a single-drug control group of arginine and chlorhexidine.
TABLE 4 inhibition of influenza A virus by tsukui gel nose drops
Figure BDA0002725383070000131
Note: -acellular lesions; + 25% cytopathic effect; , + + 50% cytopathic effect; + + + 75% cytopathy; + ++ + 100% cytopathy
TABLE 5 inhibition of influenza B virus by buna gel nasal drops
Figure BDA0002725383070000132
Note: -acellular lesions; + 25% cytopathic effect; , + + 50% cytopathic effect; + + + 75% cytopathy; + + + + + 100% cytopathy.

Claims (6)

1. The application of a compound preparation in preparing a medicament for preventing and treating respiratory tract infection diseases comprises the following raw materials:
a. arginine;
b. at least one of chlorhexidine, a chlorhexidine salt;
wherein the compound preparation is a gel preparation; the content of the raw material a is 10-50mg/ml, and the content of the raw material b is 10-50 mg/ml;
the respiratory tract infection diseases comprise diseases caused by microbial bacteria and/or virus infection, and the microbial bacteria and/or virus comprise one or more of diplococcus pneumoniae, staphylococcus aureus, escherichia coli, streptococcus type b and influenza a virus and influenza b virus.
2. The use according to claim 1, wherein the content of the raw material a is 12-50mg/ml, and the content of the raw material b is 12-50 mg/ml.
3. Use according to claim 1, wherein the chlorhexidine salt comprises a chlorhexidine organic acid salt.
4. Use according to claim 3, wherein the chlorhexidine salt comprises chlorhexidine digluconate or chlorhexidine diacetate or a mixture of both.
5. The use of claim 1, wherein the respiratory tract infection diseases comprise one or more of common cold, rhinitis, sinusitis, tonsillitis, tracheitis, bronchitis, and pneumonia.
6. The application of a compound preparation in preparing a medicament for preventing and treating vaginitis and/or cervicitis comprises the following raw materials:
a. an amino acid;
b. at least one of chlorhexidine, a chlorhexidine salt;
wherein the compound preparation is a gel preparation; the content of the raw material a is 10-50mg/ml, and the content of the raw material b is 10-50 mg/ml; the amino acid is selected from arginine; the chlorhexidine salt includes chlorhexidine digluconate or chlorhexidine diacetate or a mixture of both.
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