CN111557938A - Disinfectant for quickly damaging viral envelope - Google Patents
Disinfectant for quickly damaging viral envelope Download PDFInfo
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- CN111557938A CN111557938A CN202010438735.8A CN202010438735A CN111557938A CN 111557938 A CN111557938 A CN 111557938A CN 202010438735 A CN202010438735 A CN 202010438735A CN 111557938 A CN111557938 A CN 111557938A
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Abstract
The invention discloses a disinfectant for quickly damaging a virus envelope. The disinfectant preparation takes edible acid and cetylpyridinium chloride as main active ingredients, can cause damage to the virus envelope in a very short time, further quickly kill various envelope viruses, and can also kill and inhibit bacteria and fungi. The disinfectant can be divided into external skin disinfectant (such as hand disinfectant) and internal mucosa disinfectant (such as collutory and nasal wash) according to different adjuvants. The disinfectant can also be prepared into liquid and dry powder preparations according to different requirements. The disinfectant preparation has quick and efficient killing effect on various enveloped viruses, bacteria and fungi. In addition, the disinfectant preparation has the advantages of strong stability, no stimulation to human skin, no residue after use, easy water solubility, safety, no toxic or side effect, and suitability for disinfection of daily articles such as oral and nasal mucosa, hand and foot skin, wounds, clothes and the like.
Description
Technical Field
The invention relates to the technical field of disinfection, in particular to a disinfectant capable of quickly damaging a virus envelope.
Background
Viruses are a non-cellular organism, and typically consist of a nucleic acid and an outer protein coat that surrounds the nucleic acid, but have a portion of the virion with an outer envelope structure, which we call enveloped viruses. The envelope is mainly derived from host cell membrane, and is a lipid bilayer membrane containing protein, polysaccharide and lipid. The protein molecules on the envelope of the virus are encoded by the viral genome, often in the form of glycoproteins or lipoproteins, and can mediate the fusion of the envelope of the virus with the cell membrane or intracellular membrane of a host cell, so that the contents of the virus can be released into the host cell, and the envelope also has the function of maintaining the structural integrity of the virion. Viruses such as human acquired immunodeficiency syndrome virus (HIV), Hepatitis B Virus (HBV), Paramyxovirus (Paramyxovirus), Influenza virus (Influenza virus), Ebola virus (Ebola virus) which pose a great threat to the safety of human life health are enveloped viruses. The current common mode is to use 75% ethanol solution or high-temperature disinfection method, but the mode can only disinfect the skin in vitro, the application of the disinfectant to the mucosa in the mouth and the nose is extremely limited, and the disinfectant used for the mucosa of the skin in the market is mainly used for sterilization and has little effect on virus inactivation. The development of a skin mucosa disinfectant capable of inhibiting or inactivating viruses can greatly improve the possibility of virus infection, and is of great significance. The present inventors have made intensive studies on this for this reason, and after having paid a lot of creative efforts, have completed the present invention.
Quaternary ammonium salts have been reported to be synthesized as disinfectants as early as 1915, and the disinfectants have certain bactericidal capacity. In 1935, the bactericidal performance and bacteriostatic principle of such disinfection were reported by German scientists, and have attracted great interest. The same year is used for clinical disinfection practice. Such disinfectant drugs are then widely used for disinfecting skin mucosa, hand washing and medical instruments, and also for disinfecting appliances and equipment in various public places and in various production lines. Cetylpyridinium chloride, which is one of quaternary ammonium salt disinfectants, is also a surfactant, and can play a role in bacteriostasis and sterilization by reducing surface tension and further increasing permeability of cell walls and cell membranes. At the same time, the present study demonstrated that cetylpyridinium chloride can kill viruses directly by disrupting the viral envelope and there was no evidence of selective resistance. The cetylpyridinium chloride serving as a cationic surfactant has good surface activity in an acid medium; and easily precipitate in an alkaline medium to lose surface activity. In order to utilize the cetylpyridinium chloride to destroy the virus envelope effect to the maximum extent, the cetylpyridinium chloride and the acidic substance can be used together to kill the virus. The edible acid is used for adjusting the disinfectant to be acidic, so that the antiviral capability of the cetylpyridinium chloride can be utilized to the maximum extent, and the action time is shortened. The edible acid can be added into the disinfectant according to the standard of compound food, and has low price and cost. Therefore, the edible acid is used together with the cetylpyridinium chloride, so that the killing effect of the cetylpyridinium chloride on viruses is maximized, and the enveloped viruses can be obviously killed in a short time even at a very small concentration.
Disclosure of Invention
Specifically, the technical problems to be solved by the present invention are: provides a disinfectant preparation for quickly damaging virus envelope.
In order to solve the technical problems, the technical scheme of the invention is as follows:
the invention provides a disinfectant preparation for quickly damaging a virus envelope, which takes cetylpyridinium chloride as a main active ingredient.
The disinfectant preparation is used as a preferable scheme: the disinfectant can be divided into external skin disinfectant (such as hand disinfectant) and internal mucosa disinfectant (such as collutory and nasal wash) according to different adjuvants.
The disinfectant preparation is used as a preferable scheme: the disinfectant can also be prepared into liquid and dry powder disinfectant according to different requirements.
The disinfectant preparation is used as a preferable scheme: the composition comprises the following components in percentage by weight:
cetylpyridinium chloride 0.05-2 wt%, preferably 0.1 wt%;
0.005-0.05 wt% of citric acid, preferably 0.0125 wt%.
The disinfectant preparation adjuvant is used as a preferable scheme: including but not limited to the following: edible acid, ethanol, sodium benzoate, potassium sorbate, glycerol, poloxamer, etc.
The in vitro skin disinfection preparation is used as a preferred scheme: the composition comprises the following components in percentage by weight:
cetylpyridinium chloride 0.05-2 wt%, preferably 0.1 wt%;
0.005-0.05 wt% of citric acid, preferably 0.025 wt%;
ethanol 20-75 wt%, preferably 25 wt%;
the balance of water.
The in vivo mucosa disinfection preparation is a preferable scheme: the composition comprises the following components in percentage by weight:
cetylpyridinium chloride 0.05-2 wt%, preferably 0.05 wt%;
0.005-0.05 wt% of citric acid, preferably 0.025 wt%;
sodium benzoate 0.05-0.15 wt%, preferably 0.1 wt%;
potassium sorbate 0.01-0.05 wt%, preferably 0.04 wt%;
1-10 wt% of glycerol, preferably 5 wt%;
the balance of water.
The liquid and dry powder disinfection preparation is used as a preferred scheme: the liquid disinfectant is prepared by adding water into the effective component; the dry powder disinfectant is prepared by mixing solid effective components at a certain proportion, packaging in equal amount, and adding appropriate amount of water.
The invention provides the application of the disinfectant, and the disinfectant is suitable for disinfecting daily articles such as oral and nasal mucosa, hand and foot skin, wounds, clothes and the like so as to kill viruses and bacteria.
As a preferred embodiment, the viruses, bacteria and fungi include human acquired immunodeficiency syndrome virus (HIV), Hepatitis B Virus (HBV), Paramyxovirus (Paramyxovirus), Influenza virus (Influenza virus), Ebola virus (Ebola virus), Coronavirus (Coronavir), enteropathogenic bacteria, pyococcus, pathogenic yeast, nosocomial infection common pathogenic bacteria.
The invention provides a using method of a disinfectant, which comprises the following steps: when the skin, the wound surface and the object surface of hands and feet are disinfected, spraying or wiping the raw liquid for 2 minutes, and then cleaning; when the oral and nasal mucosa is disinfected, the original solution is used for gargling or wiping, and the action time is 1 minute.
The invention has the following advantages:
(1) the invention can quickly and efficiently damage the virus envelope, further kill various envelope viruses in a very short time, and simultaneously the disinfectant can also kill and inhibit bacteria and fungi.
(2) The disinfectant can be used for disinfecting hand and foot skins, mouth and nose mucous membranes and damaged wounds and can also be used for disinfecting daily articles such as clothes and the like. The disinfection process does not cause toxicity and irritation. The oral preparation of the invention can be used as mouthwash, and does not cause damage to skin and mucous membrane and strong irritation. (3) The disinfectant preparation has the advantages of strong stability, no stimulation to human skin, no residue after use, easy water dissolution, safety, no toxic or side effect, and can be selected from liquid agents or dry powder agents according to different requirements.
Drawings
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the present invention.
FIG. 1 shows the Western Blot detection of the expression level of HBV capsid assembly-associated functional proteins after treatment with HBVCp149 and candidate drug.
FIG. 2 shows the PCR detection of RNA levels of HBV capsid assembled related functional proteins after treatment with HBVCp149 and drug candidates.
FIG. 3 shows the quantification of HBV-Cp149 DNA fragment size after treatment with candidate drugs in HepG2.2.15 cells.
FIG. 4 is a graph of serum HBV DNA levels of mice treated with example 4 after intramuscular injection for 2 days by quantitative real-time PCR analysis. Controls were injected intramuscularly with DMSO (1: 1000 dilution).
FIG. 5 is a graph of HBV DNA levels in serum of mice treated with example 4 after intramuscular injection for 3 days by quantitative real-time PCR analysis. Controls were injected intramuscularly with DMSO (1: 1000 dilution).
Detailed Description
The technical solutions of the present invention are further illustrated below by means of specific examples, which are intended to help the better understanding of the contents of the present invention, but these specific examples do not limit the scope of the present invention in any way. The starting materials used in this embodiment are commercially available or may be obtained by methods known in the art. In this embodiment, the concentrations are given by mass percent.
Example 1
A mouthwash disinfectant for quickly damaging a viral envelope is prepared from the following components in percentage by weight:
cetylpyridinium chloride is preferably 0.1 wt%;
preferably, citric acid is 0.0125 wt%;
sodium benzoate is preferably 0.1% by weight;
potassium sorbate is preferably 0.04 wt%;
the glycerol is preferably 5 wt%;
sorbitol is preferably 5 wt%;
the zinc gluconate is preferably 0.2 wt%;
and the balance of water.
Example 2
A nasal wash disinfectant for quickly damaging a virus envelope comprises the following components in percentage by weight:
cetylpyridinium chloride is preferably 0.1 wt%;
preferably, citric acid is 0.0125 wt%;
sodium benzoate is preferably 0.1% by weight;
potassium sorbate is preferably 0.04 wt%;
the glycerol is preferably 5 wt%;
and the balance of water.
Example 3
A nasal wash disinfection dry powder preparation for rapidly damaging a virus envelope comprises the following components by weight percent:
cetylpyridinium chloride is preferably 0.1 wt%;
preferably, citric acid is 0.0125 wt%;
sodium benzoate is preferably 0.1% by weight;
potassium sorbate is preferably 0.04 wt%;
example 4
A hand and foot skin disinfectant for rapidly damaging virus envelope comprises the following components in percentage by weight:
cetylpyridinium chloride is preferably 0.1 wt%;
preferably, citric acid is 0.025 wt%;
and the balance of water.
Example 5
A hand and foot skin disinfectant for damaging virus envelope comprises the following components in percentage by weight:
cetylpyridinium chloride is preferably 0.1 wt%;
preferably, citric acid is 0.025 wt%;
ethanol is preferably 20 wt%;
and the balance of water.
Example 6
A hand and foot skin disinfection dry powder preparation for damaging virus envelope comprises the following components by weight percent:
cetylpyridinium chloride is preferably 0.1 wt%;
preferably, citric acid is 0.025 wt%;
example 7
The anti-HBV effect test of the disinfectant preparation comprises the following steps: example 4 was used as the experimental group, untreated as the normal group, and the group without citric acid addition was designated as the control group (i.e., 0.1 wt% cetylpyridinium chloride and water balance).
CPC can inhibit HBV capsid assembly in vitro, see fig. 1, fig. 2, fig. 3, fig. 4 and fig. 5.
The results of fig. 1 and 2 show that the effect of cetylpyridinium chloride on disrupting the capsular membrane is significantly enhanced in an acidic environment, and the ability to inhibit the assembly of the capsular membrane is much stronger than that of the conventional antiviral drugs.
a | b | c | d | |
Cp149 | + | + | + | + |
37℃ | - | + | + | + |
Inhibitor | - | + | Control group | Example 4 |
The results of fig. 3 show that: the Cp149DNA fragment treated in example 4 is mostly small, indicating that the DNA cannot be normally replicated or the DNA fragment is damaged, which indicates that example 4 has a good effect of damaging the viral DNA.
Example 4 inhibition of HBV infection in vivo (FIGS. 4 and 5)
The experiments of fig. 4 and 5 demonstrate that the disinfectant formulation of example 4 can achieve killing effect on HBV by inhibiting synthesis of HBV capsid in vivo and in vitro.
Example 8
The disinfectant preparation has the following effects of resisting influenza virus: example 4 was used as the experimental group.
Effective influenza virus killing concentration (EC 50) and therapeutic index for CPC.
All experiments were repeated 6 times and the same values were obtained.
Therapeutic index was calculated from CC 50/EC 50; a clinical strain of influenza A virus (H1N1pdm09) propagated from a patient sample was determined to have a CC 50 of 96 μ g/mL Clinical Isolate 40 by neutral Red analysis.
Example 9
The antibacterial effect test of the disinfectant preparation comprises the following steps: example 4 (denoted CPC) was used as the experimental group.
The average inactivation index for the novel coronaviruses and bacteria is given in the following table:
categories | Number of experiments | |
Staphylococcus aureus | ||
3 | >99% | |
Escherichia coli | 3 | >99% |
Candida albicans | 3 | >99% |
From the above results, the disinfectant provided by the invention has a fast and efficient killing effect on pathogenic bacteria and fungi.
Example 10
Acute eye irritation test
Experimental groups: EXAMPLE one (stock solution)
Experimental animals: white rabbit, acute eye irritation test, three rabbits
The experimental method comprises the following steps:
1. the rabbit was fixed, the lower eyelid of the right eye of the rabbit was gently pulled open, 0.5ml (100mg) of the test sample was administered to the conjunctival sac to allow the upper and lower eye curtains to be passively closed for 30 seconds to prevent the test sample from being lost, and the untreated other eye was used as a self-control or as a drop vehicle.
2. If the test result shows that the tested object has irritation, and if the test result shows that the tested object has irritation, the tested object is dripped into the eyes of the domestic rabbits for 30 seconds, and the tested object is washed for at least 30 seconds by using water flow which has enough quantity and high flow speed and can not cause the injury of the eyes of the animals.
Clinical examination and scoring:
the eyes of the animals were examined 1, 24, 48, 72h and 4d and 7d after instillation of the test substance. If no stimulus response occurs for 72 hours, the test is terminated. If the cornea is found to be involved or other eye irritation is found, the examination is made. If no stimulus response occurs for 72 hours, the test is terminated. If corneal involvement or other ocular irritation is detected and the patient does not recover after 7 days, the observation time is prolonged to determine the reversibility or irreversibility of the lesion, which is generally not more than 21 days. In addition to corneal, iris, conjunctival observations, other damaging effects should be recorded and reported. After 24h observation and recording, changes in the cornea and iris can be examined using 2% sodium fluorescein solution or slit lamp, loupe, etc.
From the above results, the disinfectant provided by the invention has less irritation and side effects, and the components are simpler, so that the side effects and irritation of certain people are avoided as much as possible.
Although the invention has been described with reference to a preferred embodiment, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention.
Claims (15)
1. A disinfectant preparation for quickly damaging a viral envelope is characterized in that: the disinfectant preparation takes edible acid and cetylpyridinium chloride as main active ingredients, can cause damage to the virus envelope in a very short time, further quickly kill various envelope viruses, and can also kill and inhibit bacteria and fungi.
2. Use according to claim 1, characterized in that: the cetylpyridinium chloride is one of quaternary ammonium salt disinfectants and is also a surfactant, is named cetyl pyridinium chloride in a Chinese name, and has a molecular formula of C21H38ClN ● H2O.
3. Use according to claim 1, characterized in that: cetylpyridinium chloride can kill enveloped viruses directly by disrupting the viral envelope and there is no evidence of selective resistance. By matching with edible acid, the disinfectant is in a weakly acidic environment, so that the killing effect of cetylpyridinium chloride on viruses and bacteria is greatly enhanced.
4. The edible acid of claim 3, wherein: the edible acid as a food additive mainly comprises: citric acid (also called citric acid), malic acid, gluconic acid, lactic acid, tartaric acid, phosphoric acid, fumaric acid, adipic acid, succinic acid, etc.
5. Use according to claim 3, characterized in that: a weakly acidic disinfectant or pharmaceutical preparation containing cetylpyridinium chloride as main ingredient can inhibit the activity of viruses, especially enveloped viruses, most pathogenic bacteria or fungi in a short time.
6. A sanitising formulation or pharmaceutical product according to claim 5 wherein: the composition comprises the following components in percentage by weight:
cetylpyridinium chloride 0.05-2 wt%, preferably 0.1 wt%;
0.005-0.05 wt% of citric acid, preferably 0.0125 wt%.
7. A sanitising formulation or pharmaceutical product according to claim 5 wherein: the disinfectant can be divided into external skin disinfectant (such as hand disinfectant), internal mucosa disinfectant (such as collutory and nasal wash) or corresponding medicines according to different additives, and can be added into washing products such as laundry detergent and liquid detergent for killing bacteria, fungi and viruses in daily necessities.
8. A sanitising formulation or pharmaceutical product according to claim 5 wherein: the disinfectant can also be prepared into liquid and dry powder disinfectant according to different requirements.
9. The enveloped virus, bacterium and fungus of claim 5 characterized in that: including human acquired immunodeficiency syndrome virus (HIV), Hepatitis B Virus (HBV), Paramyxovirus (Paramyxovirus), Influenza virus (Influenza virus), Ebola virus (Ebola virus), enteropathogenic bacteria, pyococcus, pathogenic yeast, common pathogenic bacteria for hospital infection, etc.
10. A sanitising formulation or pharmaceutical adjuvant according to claim 7 wherein: including but not limited to the following: ethanol, sodium benzoate, potassium sorbate, glycerol, poloxamer, etc.
11. The in vitro skin sanitizing formulation of claim 7, wherein: the composition comprises the following components in percentage by weight:
cetylpyridinium chloride 0.05-2 wt%, preferably 0.1 wt%;
0.005-0.05 wt% of citric acid, preferably 0.025 wt%;
ethanol 20-75 wt%, preferably 0.025 wt%;
the balance of water.
12. The in vivo mucosal disinfecting formulation of claim 7, wherein: the composition comprises the following components in percentage by weight:
cetylpyridinium chloride 0.05-2 wt%, preferably 0.05 wt%;
0.005-0.05 wt% of citric acid, preferably 0.0125 wt%;
sodium benzoate 0.05-0.15 wt%, preferably 0.1 wt%;
potassium sorbate 0.01-0.05 wt%, preferably 0.04 wt%;
1-10 wt% of glycerol, preferably 5 wt%;
the balance of water.
13. A liquid and dry powder sanitizing formulation as set forth in claim 8, wherein: the liquid disinfectant is prepared by adding water into the effective component; the dry powder disinfectant is prepared by mixing solid effective components at a certain proportion, packaging in equal amount, and adding appropriate amount of water.
14. Use of a sanitising agent or a medicament as claimed in claim 3 wherein: is suitable for sterilizing daily articles such as oral mucosa, nasal mucosa, hand and foot skin, wound and clothes, etc., to kill bacteria, fungi and viruses.
15. Use according to claim 14, characterized in that: when the skin, the wound surface and the object surface of hands and feet are disinfected, spraying or wiping the raw liquid for 2 minutes, and then cleaning; when the oral and nasal mucosa is disinfected, the original solution is used for gargling or wiping, and the action time is 1 minute.
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