CN114097832A - Long-acting inactivated virus spray and preparation method thereof - Google Patents
Long-acting inactivated virus spray and preparation method thereof Download PDFInfo
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- CN114097832A CN114097832A CN202111444064.7A CN202111444064A CN114097832A CN 114097832 A CN114097832 A CN 114097832A CN 202111444064 A CN202111444064 A CN 202111444064A CN 114097832 A CN114097832 A CN 114097832A
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- honeysuckle
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- fructus cnidii
- ethanol
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- VTAJIXDZFCRWBR-UHFFFAOYSA-N Licoricesaponin B2 Natural products C1C(C2C(C3(CCC4(C)CCC(C)(CC4C3=CC2)C(O)=O)C)(C)CC2)(C)C2C(C)(C)CC1OC1OC(C(O)=O)C(O)C(O)C1OC1OC(C(O)=O)C(O)C(O)C1O VTAJIXDZFCRWBR-UHFFFAOYSA-N 0.000 description 1
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- 239000001685 glycyrrhizic acid Substances 0.000 description 1
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- LPLVUJXQOOQHMX-QWBHMCJMSA-N glycyrrhizinic acid Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@H](O[C@@H]1O[C@@H]1C([C@H]2[C@]([C@@H]3[C@@]([C@@]4(CC[C@@]5(C)CC[C@@](C)(C[C@H]5C4=CC3=O)C(O)=O)C)(C)CC2)(C)CC1)(C)C)C(O)=O)[C@@H]1O[C@H](C(O)=O)[C@@H](O)[C@H](O)[C@H]1O LPLVUJXQOOQHMX-QWBHMCJMSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/10—Apiaceae or Umbelliferae [Carrot family], e.g. parsley, caraway, dill, lovage, fennel or snakebed
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
- A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
- A01N25/06—Aerosols
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Agronomy & Crop Science (AREA)
- Plant Pathology (AREA)
- Environmental Sciences (AREA)
- Zoology (AREA)
- Mycology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Toxicology (AREA)
- Pest Control & Pesticides (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Medicinal Preparation (AREA)
- Cosmetics (AREA)
Abstract
The invention provides a long-acting inactivated virus spray and a preparation method thereof, belonging to the technical field of disinfection and antibiosis; according to the invention, by adjusting the proportion of honeysuckle, forsythia and fructus cnidii and the proportion of 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester and fucoidan, the synergistic effect can be obviously exerted, the antiviral effect is obvious, the viruses can be effectively inhibited from invading and harming human bodies, the killing effect on new coronavirus is obvious, and the effect is more durable. The spray is used in a spray form, is used for disinfecting ambient air, can effectively kill viruses in a short time, is nontoxic and harmless to human bodies, has the advantages of naturalness, safety and mild pesticide effect, has no toxic or side effect, is simple in preparation method and low in cost, and is beneficial to large-scale production.
Description
Technical Field
The invention belongs to the technical field of disinfection and antibiosis, and particularly relates to a long-acting inactivated virus spray and a preparation method thereof.
Background
Viral pneumonia is common in winter and spring, can be sporadic or fulminant, and is clinically manifested as fever, general aching pain, small part of the body with dyspnea, and lung infiltration shadow. Viral pneumonia is associated with the virulence of the virus, the route of infection, and the age and immune status of the host. Viruses causing viral pneumonia are common to influenza viruses, and others are parainfluenza viruses, cytomegaloviruses, adenoviruses, rhinoviruses, coronaviruses, and the like. The diagnosis depends on etiology examination, including virus isolation, serological examination, and detection of viral antigens and nucleic acids. Coronavirus, RNA virus, divided into four genera of alpha, beta, gamma and delta, wherein the novel coronavirus (SARS-CoV-2) is a novel coronavirus of beta genus, and has envelope, round or elliptic net shape particle, usually polymorphism, and diameter of 60-140 nm. The S protein is one of the main proteins of the novel coronavirus, the coding gene of the S protein is used for virus typing, and the respiratory epithelial cells of a human are infected through a molecular mechanism of interaction of the S-protein and human ACE2, so that the novel coronavirus has strong infectivity on the human. This pulmonary infection caused by the novel coronavirus is called novel coronavirus pneumonia. The N protein wraps the viral genome and can be used as a diagnostic antigen.
The main transmission routes of the novel coronavirus are respiratory droplet transmission (sneezing, coughing and talking droplets of a patient and infection caused by the fact that exhaled air is directly inhaled in a short distance) and contact transmission (the droplets are deposited on the surface of an object and contact with mucous membranes such as oral cavities, nasal cavities, eyes and the like after contacting with polluted hands to cause infection). Epidemiological investigation shows that many cases can be traced to the close contact with the diagnosed cases.
Alcohol, chemical products containing chlorine components and some disinfection compositions are widely applied to inactivation of new coronavirus, but the alcohol has extremely strong volatility and short action time, 75 percent of alcohol is flammable and explosive substances of the alcohol, and large-area, high-concentration and frequent spraying easily causes fire hazard; the disinfectant containing chloride ions has unacceptable smell and irritation to the skin, and has pathogenic risk when contacting the skin for a long time. Patent CN112266646A discloses a long-acting slow-release antibacterial and disinfectant composition, which comprises the following components in percentage by mass: 0.05-95% of polymer; 0.001-10% of bactericidal disinfectant; 0.01-90% of other additives, the composition can form an organic coating, the coating has excellent affinity to human skin, has good adhesion to various surfaces, can be cured at room temperature, and slowly releases various sterilizing and disinfecting drugs; the coating has a self-cleaning function, and can keep the effects of long-lasting sterilization and killing new coronavirus on the surface of the coating. Patent CN113144177A discloses an oropharyngeal surface and hand surface sterilization and antiviral spray, which is prepared from lysozyme, tetrapyrachlor, glycyrrhizic acid, acesulfame, menthol and hydrogen-rich water, and has the characteristics of broad-spectrum antivirus, extremely strong permeability, high action speed, good stability, low toxicity, no corrosiveness, small irritation and good environmental protection property, and particularly, the spray is easy to form a layer of 'invisible protective film' on the oral mucosa, the face and the skin, particularly the hand surface, can effectively prevent the infection of bacteria and viruses, and has the effects of bacteriostasis and antivirus for a long time. In the prior art, the spray for inactivating the new coronavirus has fewer types, short action time and poorer safety.
Therefore, the development of a long-acting, effective, safe and economic spray for inactivating the new coronavirus is an urgent problem to be solved.
Disclosure of Invention
The purpose of the invention is as follows: in order to overcome the defects in the prior art, the invention provides a long-acting inactivated virus spray and a preparation method thereof. The spray has good effect of killing new coronavirus, long action time and high safety.
The technical scheme is as follows: in order to achieve the purpose, the technical scheme of the invention is as follows:
in one aspect, the present invention provides an inactivated virus spray, comprising: honeysuckle, forsythia, fructus cnidii, 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol.
Specifically, the spray comprises the following components in parts by weight: 10-70 parts of honeysuckle, 5-40 parts of fructus forsythiae, 5-40 parts of fructus cnidii, 0.1-5 parts of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 0.1-5 parts of polyglycerol fatty acid ester, 0.01-1 part of fucoidan, 1-20 parts of ethanol and 1-20 parts of glycerol.
Further specifically, the spray comprises the following components in parts by weight: 30-50 parts of honeysuckle, 10-30 parts of fructus forsythiae, 10-30 parts of fructus cnidii, 0.5-2 parts of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 0.5-2 parts of polyglycerol fatty acid ester, 0.05-0.5 part of fucoidan, 5-15 parts of ethanol and 5-15 parts of glycerol.
Specifically, the weight ratio of the honeysuckle to the fructus forsythiae to the fructus cnidii is 1-5:1:1, and preferably 2:1: 1.
Specifically, the weight ratio of the 3- [ trimethoxysilyl ] propyl dimethyloctadecyl ammonium chloride, the polyglycerol fatty acid ester and the fucoidan sulfate is 1:1:0.05-0.5, and preferably 1:1: 0.1.
Further specifically, the spray comprises the following components in parts by weight: 40 parts of honeysuckle, 20 parts of forsythia, 20 parts of fructus cnidii, 1 part of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 1 part of polyglycerol fatty acid ester, 0.1 part of fucoidan, 10 parts of ethanol and 10 parts of glycerol.
On the other hand, the invention provides a preparation method of the inactivated virus spray, which comprises the following steps:
(1) taking honeysuckle, fructus forsythiae and fructus cnidii according to weight fractions, putting the honeysuckle, the fructus forsythiae and the fructus cnidii into a grinder, fully grinding the honeysuckle, the fructus forsythiae and the fructus cnidii, and sieving the ground honeysuckle, the fructus forsythiae and the fructus cnidii to obtain honeysuckle powder, fructus cnidii powder respectively;
(2) heating and refluxing the honeysuckle powder, the forsythia powder and the fructus cnidii powder prepared in the step (1) by using ethanol to prepare honeysuckle alcohol extract, forsythia alcohol extract and fructus cnidii alcohol extract;
(3) and (3) mixing the honeysuckle alcohol extract, the forsythia alcohol extract and the fructus cnidii alcohol extract prepared in the step (2) with 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol according to parts by weight, and uniformly stirring to obtain the inactivated virus spray.
Specifically, the preparation method of the alcohol extract in the step (2) comprises the following steps: extracting the powder with 85-90% ethanol under reflux for 1-3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 2-4 times to obtain ethanol extract.
Further specifically, the preparation method of the alcohol extract in the step (2) comprises the following steps: extracting the powder with 85% ethanol under reflux for 3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 3 times to obtain ethanol extract.
In another aspect, the invention provides the application of the spray in preparing a medicine for inactivating the new coronavirus.
Has the advantages that: compared with the prior art, the invention has the advantages that:
the invention provides an inactivated new corona virus spray, which is characterized in that all components are mutually compatible and optimally combined, the proportion of honeysuckle, forsythia and fructus cnidii and the proportion of 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester and fucoidan sulfate are adjusted, so that the obvious synergistic effect can be exerted, the antiviral effect is obvious, the virus invasion and harm to human bodies can be effectively inhibited, the killing effect on new corona viruses is obvious, and the effect is more durable. The spray is used in a spray form, is used for disinfecting ambient air, can effectively kill viruses in a short time, is nontoxic and harmless to human bodies, has the advantages of naturalness, safety and mild pesticide effect, has no toxic or side effect, is simple in preparation method and low in cost, and is beneficial to large-scale production.
Detailed Description
The present invention will be further illustrated in detail with reference to the following specific examples, which are not intended to limit the present invention but are merely illustrative thereof. The experimental methods used in the following examples are not specifically described, and the materials, reagents and the like used in the following examples are generally commercially available under the usual conditions without specific descriptions.
The examples, where no specific techniques or conditions are indicated, are carried out according to the techniques or conditions described in the literature of the art (for example, see J. SammBruk et al, molecular cloning, A laboratory Manual, third edition, scientific Press, ed. by Huang Pe, et al) or according to the instructions of the product. The experimental methods in the following examples are conventional methods unless otherwise specified, and the bacterial species, drugs, reagents and the like used in the following examples are commercially available without otherwise specified.
The invention adopts independent repeated experiments, and the experiments are repeated for 3 times. The experimental data are analyzed by SPSS18.0 statistical analysis software, the measured data are expressed by mean +/-standard deviation, the mean of the two samples is compared, and after the normality test and the homogeneity test of the variance, the difference is suggested to have statistical significance by adopting t or t' test, wherein P is less than 0.05.
Example 1
An inactivated neo-coronavirus spray, said inactivated neo-coronavirus spray comprising:
40 parts of honeysuckle, 20 parts of forsythia, 20 parts of fructus cnidii, 1 part of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 1 part of polyglycerol fatty acid ester, 0.1 part of fucoidan, 10 parts of ethanol and 10 parts of glycerol.
The preparation method of the spray comprises the following steps:
(1) taking honeysuckle, fructus forsythiae and fructus cnidii according to weight fractions, putting the honeysuckle, the fructus forsythiae and the fructus cnidii into a grinder, fully grinding the honeysuckle, the fructus forsythiae and the fructus cnidii, and sieving the ground honeysuckle, the fructus forsythiae and the fructus cnidii to obtain honeysuckle powder, fructus cnidii powder respectively;
(2) and (2) carrying out heating reflux extraction on the honeysuckle powder, the forsythia powder and the fructus cnidii powder prepared in the step (1) by adopting ethanol to prepare a honeysuckle alcohol extract, a forsythia alcohol extract and a fructus cnidii alcohol extract, wherein the specific ethanol heating reflux extraction step is as follows: extracting the powder with 85% ethanol under reflux for 3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 3 times to obtain ethanol extract;
(3) and (3) mixing the honeysuckle alcohol extract, the forsythia alcohol extract and the fructus cnidii alcohol extract prepared in the step (2) with 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol according to parts by weight, and uniformly stirring to obtain the inactivated new coronavirus spray.
Example 2
An inactivated neo-coronavirus spray, said inactivated neo-coronavirus spray comprising:
50 parts of honeysuckle, 10 parts of forsythia, 10 parts of fructus cnidii, 2 parts of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 2 parts of polyglycerol fatty acid ester, 0.1 part of fucoidan, 15 parts of ethanol and 5 parts of glycerol.
The preparation method of the spray comprises the following steps:
(1) taking honeysuckle, fructus forsythiae and fructus cnidii according to weight fractions, putting the honeysuckle, the fructus forsythiae and the fructus cnidii into a grinder, fully grinding the honeysuckle, the fructus forsythiae and the fructus cnidii, and sieving the ground honeysuckle, the fructus forsythiae and the fructus cnidii to obtain honeysuckle powder, fructus cnidii powder respectively;
(2) and (2) carrying out heating reflux extraction on the honeysuckle powder, the forsythia powder and the fructus cnidii powder prepared in the step (1) by adopting ethanol to prepare a honeysuckle alcohol extract, a forsythia alcohol extract and a fructus cnidii alcohol extract, wherein the specific ethanol heating reflux extraction step is as follows: extracting the powder with 85% ethanol under reflux for 3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 3 times to obtain ethanol extract;
(3) and (3) mixing the honeysuckle alcohol extract, the forsythia alcohol extract and the fructus cnidii alcohol extract prepared in the step (2) with 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol according to parts by weight, and uniformly stirring to obtain the inactivated new coronavirus spray.
Example 3
An inactivated neo-coronavirus spray, said inactivated neo-coronavirus spray comprising:
30 parts of honeysuckle, 30 parts of weeping forsythia, 30 parts of common cnidium fruit, 0.5 part of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 0.5 part of polyglycerol fatty acid ester, 0.25 part of fucoidan, 5 parts of ethanol and 15 parts of glycerol.
The preparation method of the spray comprises the following steps:
(1) taking honeysuckle, fructus forsythiae and fructus cnidii according to weight fractions, putting the honeysuckle, the fructus forsythiae and the fructus cnidii into a grinder, fully grinding the honeysuckle, the fructus forsythiae and the fructus cnidii, and sieving the ground honeysuckle, the fructus forsythiae and the fructus cnidii to obtain honeysuckle powder, fructus cnidii powder respectively;
(2) and (2) carrying out heating reflux extraction on the honeysuckle powder, the forsythia powder and the fructus cnidii powder prepared in the step (1) by adopting ethanol to prepare a honeysuckle alcohol extract, a forsythia alcohol extract and a fructus cnidii alcohol extract, wherein the specific ethanol heating reflux extraction step is as follows: extracting the powder with 85% ethanol under reflux for 3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 3 times to obtain ethanol extract;
(3) and (3) mixing the honeysuckle alcohol extract, the forsythia alcohol extract and the fructus cnidii alcohol extract prepared in the step (2) with 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol according to parts by weight, and uniformly stirring to obtain the inactivated new coronavirus spray.
Example 4
An inactivated neo-coronavirus spray, said inactivated neo-coronavirus spray comprising:
60 parts of honeysuckle, 5 parts of forsythia, 5 parts of fructus cnidii, 3 parts of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 3 parts of polyglycerol fatty acid ester, 2 parts of fucoidan, 20 parts of ethanol and 1 part of glycerol.
The preparation method of the spray comprises the following steps:
(1) taking honeysuckle, fructus forsythiae and fructus cnidii according to weight fractions, putting the honeysuckle, the fructus forsythiae and the fructus cnidii into a grinder, fully grinding the honeysuckle, the fructus forsythiae and the fructus cnidii, and sieving the ground honeysuckle, the fructus forsythiae and the fructus cnidii to obtain honeysuckle powder, fructus cnidii powder respectively;
(2) and (2) carrying out heating reflux extraction on the honeysuckle powder, the forsythia powder and the fructus cnidii powder prepared in the step (1) by adopting ethanol to prepare a honeysuckle alcohol extract, a forsythia alcohol extract and a fructus cnidii alcohol extract, wherein the specific ethanol heating reflux extraction step is as follows: extracting the powder with 85% ethanol under reflux for 3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 3 times to obtain ethanol extract;
(3) and (3) mixing the honeysuckle alcohol extract, the forsythia alcohol extract and the fructus cnidii alcohol extract prepared in the step (2) with 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol according to parts by weight, and uniformly stirring to obtain the inactivated new coronavirus spray.
Example 5
An inactivated neo-coronavirus spray, said inactivated neo-coronavirus spray comprising:
20 parts of honeysuckle, 40 parts of forsythia, 40 parts of common cnidium fruit, 0.5 part of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 0.5 part of polyglycerol fatty acid ester, 0.01 part of fucoidan, 1 part of ethanol and 20 parts of glycerol.
The preparation method of the spray comprises the following steps:
(1) taking honeysuckle, fructus forsythiae and fructus cnidii according to weight fractions, putting the honeysuckle, the fructus forsythiae and the fructus cnidii into a grinder, fully grinding the honeysuckle, the fructus forsythiae and the fructus cnidii, and sieving the ground honeysuckle, the fructus forsythiae and the fructus cnidii to obtain honeysuckle powder, fructus cnidii powder respectively;
(2) and (2) carrying out heating reflux extraction on the honeysuckle powder, the forsythia powder and the fructus cnidii powder prepared in the step (1) by adopting ethanol to prepare a honeysuckle alcohol extract, a forsythia alcohol extract and a fructus cnidii alcohol extract, wherein the specific ethanol heating reflux extraction step is as follows: extracting the powder with 85% ethanol under reflux for 3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 3 times to obtain ethanol extract;
(3) and (3) mixing the honeysuckle alcohol extract, the forsythia alcohol extract and the fructus cnidii alcohol extract prepared in the step (2) with 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol according to parts by weight, and uniformly stirring to obtain the inactivated new coronavirus spray.
EXAMPLE 1 cytotoxicity test
1. Experimental methods
The specific experimental steps are as follows:
(1) by 1 × 105Concentration per well, human embryonic kidney cells 293(HEK293 cells) were seeded into 96-well plates and 100. mu.L DMEM medium was added thereto at 37 ℃ with 5% CO2Culturing until a monolayer of cells is formed;
(2) discarding culture solution, washing with PBS for 2 times, diluting the spray prepared in examples 1-5 of the present invention with DMEM in gradient, inoculating on cells at a concentration of 100 μ L/well with more than 4 multiple wells, setting the same concentration control group, 37 deg.C, and 5% CO2Culturing for 48 h;
(3) mu.L of MTT solution (2.5mg/mL) was added to each well at 37 ℃ with 5% CO2Incubating for 4 h; discarding the supernatant, adding 120 mu LDMSO/well, and oscillating for 20 min;
(4) measuring absorbance (OD) of 490nm wavelength of each well on a multifunctional reader, and calculating 50% toxicity concentration by Reed-Muench methodHalf the Toxic Concentration (TC)50)。
2. Results of the experiment
The results of the measurements are shown in Table 1 below.
TABLE 1 cytotoxicity assay results
| Group of | TC50(μg/mL) |
| Example 1 | 116.28 |
| Example 2 | 115.56 |
| Example 3 | 115.71 |
| Example 4 | 113.39 |
| Example 5 | 114.35 |
As shown in Table 1, the spray prepared by the invention has no cytotoxicity.
Experimental example 2 antiviral Performance test
1. Experimental methods
The specific experimental steps are as follows:
(1) HEK293 cells were plated at 4.5X 104The cells/well concentration was added to a 96-well plate in DMEM containing 1% fetal bovine serum at 37 ℃ with 5% CO2Culturing for 24 h;
(2) discard the supernatant and add 100TCID to each well separately502019-nCoV virus liquid, 5% CO at 37 DEG C2Adsorbing for 2 h;
(3) discarding supernatant, washing cells with PBS, performing gradient dilution on the spray prepared in examples 1-5 of the invention with DMEM, respectively adding into a well plate to obtain an experimental group, adding DMEM with the same amount into a blank control group, adding 5% CO at 37 deg.C2Incubating for 72h, and collecting virus supernatant;
(4) HEK293 cells were plated at 2X 106The cells/well were inoculated into 12-well plates at 37 ℃ with 5% CO2Culturing for 24 h;
(5) diluting the virus supernatant collected in step (3) by 10 times gradient, inoculating to HEK293 cells, and culturing at 37 deg.C with 5% CO2Adsorbing for 2 h;
(6) the supernatant was discarded and the cells were washed with PBS, 2mL of 1% (w/v) methylcellulose-DMEM overlay was added, 5% CO at 37 ℃2Culturing for 72 h;
(7) removing the methylcellulose-DMEM covering, washing the cells 2 times with PBS, and fixing the cells with 10% formaldehyde for 30min at room temperature;
(8) the supernatant was discarded, the cells were washed with PBS 2 times, stained with 0.5% crystal violet for 5min at room temperature, washed with deionized water 2 times, dried and the plaques were counted, and the 50% virus-inhibited drug concentration was calculated as the half-inhibitory concentration of the drug by the Reed-Muench method (IC 50).
2. Results of the experiment
The results of the measurements are shown in Table 2 below.
TABLE 2 results of antiviral Performance test
| Group of | IC50(μg/mL) |
| Example 1 | 12.16 |
| Example 2 | 12.97 |
| Example 3 | 13.34 |
| Example 4 | 17.21 |
| Example 5 | 18.33 |
As shown in Table 2, the spray prepared by the invention has better antiviral activity.
Experimental example 3 time limit verification of inactivated Virus
The experiment for inactivation of the new coronavirus 2019-nCoV was carried out with reference to Disinfection Specification, second part-2.1.1.7 (2002 edition).
The spray of the embodiment 1-5 of the invention is subjected to inactivation detection of the new coronavirus 2019-nCoV, the spray is subjected to detection after reacting with the new coronavirus 2019-nCoV for 2h, 1d, 7d and 14d, and blank control is set in the experiment. The results of the measurements are shown in Table 3 below.
TABLE 3 antiviral time limit test results
| Time | Example 1 | Example 2 | Example 3 | Example 4 | Example 5 |
| 2h | 99.99 | 99.96 | 99.95 | 99.92 | 99.90 |
| 1d | 99.97 | 99.89 | 99.82 | 99.77 | 99.68 |
| 7d | 99.91 | 99.73 | 99.70 | 99.41 | 99.22 |
| 14d | 99.53 | 99.30 | 99.25 | 99.05 | 89.76 |
As can be seen from the above table 3, the application can achieve long-acting killing of the new coronavirus 2019-nCoV by adjusting the content and the proportion of each component in the spray, and the virus inactivation time limit is longer.
The above description is only of the preferred embodiments of the present invention, and it should be noted that: it will be apparent to those skilled in the art that various modifications and adaptations can be made without departing from the principles of the invention and these are intended to be within the scope of the invention.
Claims (10)
1. An inactivated virus spray is characterized in that: the spray comprises the following components: honeysuckle, forsythia, fructus cnidii, 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol.
2. The spray according to claim 1, wherein: the spray comprises the following components in parts by weight: 10-70 parts of honeysuckle, 5-40 parts of fructus forsythiae, 5-40 parts of fructus cnidii, 0.1-5 parts of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 0.1-5 parts of polyglycerol fatty acid ester, 0.01-1 part of fucoidan, 1-20 parts of ethanol and 1-20 parts of glycerol.
3. The spray according to claim 2, wherein: the spray comprises the following components in parts by weight: 30-50 parts of honeysuckle, 10-30 parts of fructus forsythiae, 10-30 parts of fructus cnidii, 0.5-2 parts of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 0.5-2 parts of polyglycerol fatty acid ester, 0.05-0.5 part of fucoidan, 5-15 parts of ethanol and 5-15 parts of glycerol.
4. A spray according to claim 3, wherein: the weight ratio of the honeysuckle to the fructus forsythiae to the fructus cnidii is 1-5:1: 1; the weight ratio of the 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, the polyglycerol fatty acid ester and the fucoidan sulfate is 1:1: 0.05-0.5.
5. The spray according to claim 4, wherein: the weight ratio of the honeysuckle to the fructus forsythiae to the fructus cnidii is 2:1: 1; the weight ratio of the 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, the polyglycerol fatty acid ester and the fucoidan sulfate is 1:1: 0.1.
6. The spray according to claim 5, wherein: the spray comprises the following components in parts by weight: 40 parts of honeysuckle, 20 parts of forsythia, 20 parts of fructus cnidii, 1 part of 3- [ trimethoxy silicon group ] propyl dimethyl octadecyl ammonium chloride, 1 part of polyglycerol fatty acid ester, 0.1 part of fucoidan, 10 parts of ethanol and 10 parts of glycerol.
7. The method for preparing the inactivated virus spray according to any one of claims 1 to 6, wherein the method comprises the following steps: the preparation method comprises the following steps:
(1) taking honeysuckle, fructus forsythiae and fructus cnidii according to weight fractions, putting the honeysuckle, the fructus forsythiae and the fructus cnidii into a grinder, fully grinding the honeysuckle, the fructus forsythiae and the fructus cnidii, and sieving the ground honeysuckle, the fructus forsythiae and the fructus cnidii to obtain honeysuckle powder, fructus cnidii powder respectively;
(2) heating and refluxing the honeysuckle powder, the forsythia powder and the fructus cnidii powder prepared in the step (1) by using ethanol to prepare honeysuckle alcohol extract, forsythia alcohol extract and fructus cnidii alcohol extract;
(3) and (3) mixing the honeysuckle alcohol extract, the forsythia alcohol extract and the fructus cnidii alcohol extract prepared in the step (2) with 3- [ trimethoxysilyl ] propyl dimethyl octadecyl ammonium chloride, polyglycerol fatty acid ester, fucoidan, ethanol and glycerol according to parts by weight, and uniformly stirring to obtain the inactivated virus spray.
8. The method of claim 7, wherein: the preparation method of the alcohol extract in the step (2) comprises the following steps: extracting the powder with 85-90% ethanol under reflux for 1-3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 2-4 times to obtain ethanol extract.
9. The method of claim 8, wherein: the preparation method of the alcohol extract in the step (2) comprises the following steps: extracting the powder with 85% ethanol under reflux for 3 times, recovering solvent under reduced pressure to obtain extract, dispersing the extract in distilled water, and extracting with ethyl acetate for 3 times to obtain ethanol extract.
10. Use of a spray according to any one of claims 1 to 6 for the preparation of a medicament for inactivating a new coronavirus.
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