CN1548152A - Treating and preventing effect of lysozyme on SARS - Google Patents
Treating and preventing effect of lysozyme on SARS Download PDFInfo
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- CN1548152A CN1548152A CNA031266010A CN03126601A CN1548152A CN 1548152 A CN1548152 A CN 1548152A CN A031266010 A CNA031266010 A CN A031266010A CN 03126601 A CN03126601 A CN 03126601A CN 1548152 A CN1548152 A CN 1548152A
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Abstract
The present invention is use of lysozyme as medicine and disinfectant. Lysozyme is used in preventing and treating SARS.
Description
● invention field
The present invention adopts lysozyme as medicine and disinfectant applications, is used for the treatment of SARS treatment of diseases and prevention.
● background technology
Severe acute respiratory syndrome (Severe Acute Respiratory Syndrome, SARS), this is meant popular a kind of respiratory infectious disease in the recent period, it at first takes place popular, popular 20 a plurality of countries and regions at present in area, China Guangdong.The route of infection are mainly by short distance droplet transmission, contact patient respiration road secretions and close contact transmission.The general susceptible of crowd, medical personnel are high-risk group of primary disease.The clinical pneumonia that mainly shows as, classical symptom is (>38 ℃), dry cough, rapid breathing or the dyspnea of having a high fever.The mortality rate of SARS sufferer is about 4%.World Health Organization (WHO) is named as severe acute respiratory syndrome on March 15th, 2003, this disease symptoms shows as and is about its incubation period between two to 12 days, usually at four to five days.Onset is anxious, be first symptom with heating, and body temperature is general>and 38 ℃, fear of cold is arranged by chance; Can be with headache, joint aches, muscular soreness, weak, diarrhoea; Normal other symptom of no upper respiratory tract; Cough can be arranged, mostly be dry cough, few expectorant, idol has sputum with blood streaks; Can have uncomfortable in chestly, severe patient occur to breathe quickens, tachypnea, or obviously respiratory distress.Prevention and treatment for this disease do not have the vaccine prevention atypical pneumonia at present; There are not specific medicament and Therapeutic Method, but behind supportive treatment and symptomatic treatment timely, the most humans rehabilitation of can fully recovering.World Health Organization (WHO) announces that on April 16th, 2003 causing a disease of SARS was a kind of new coronavirus originally, the called after SARS virus.
The status of SARS virus on taxonomy is: ssRNA positive strand virus, noDNA stage, Nidovirales, cornaviridae, cornavirus, SARScornavirus.
SARS virus is the single strand RNA virus of normal chain, duplicates the intermediate without DNA, uses standard cipher.In GeneBank, have the whole genome sequence and the note of Urbani, CUHK-W1,14 strains such as HKU-39849, BJ01 at present.Coronavirus is a kind of ssRNA (+) virus, and under Electronic Speculum, virion is irregular shape, and diameter is about 60 ~ 220nm, and peplos is arranged, and there is quincuncial peplomer on its surface, and shape such as Phaleria macrocarpa are so be called coronavirus (coronavirus).The granule center is indefinite form under the negative staining Electronic Speculum, nucleocapsid is rarefaction.Two kinds of glycoproteins are arranged: S albumen on the peplos: spike glycoprotein is main antigen, with receptors bind, makes cell fusion.M albumen: stride film, participate in peplos and form.In some subclass, also has the third glycoprotein, HE---hemagglutinin esterase.Geneome RNA combines with alkaline phosphatase albumen N.The human corona virus belongs to OC43 and two antigenic types of 229E respectively, and it is the cause of disease that causes human upper respiratory tract infection, often causes adult's common cold.
In to SARS treatment of diseases and prevention process, it is the respiratory disorder that coronavirus infection causes for the preliminary proof of etiology research.Clinical observation is found: dead patient person in middle and old age case, chronically ill man are many, the resistance of individual patients plays a significantly greater role in therapeutic process, the application of various antibiotic, antiviral drugs means is not subjected to Expected Results, makes the application of clinical medicine be restricted.The medicine that searching has different antivirus action mechanism becomes a major issue.
Lysozyme is a clinical application for many years, machine-processed different with existing antibiotic, antiviral drugs, its antibiosis and antiviral functions mechanism is unique: 1], the lysozyme antibiotic effect is by the β-1 between the N-acetylglucosamine in the cell wall that destroys antibacterial, 4 glycosidic bonds, make the insoluble mucopolysaccharide of cell wall resolve into the solubility glucosides, cause cell wall rupture, cellular content is overflowed and is made bacterolysis.2], lysozyme can also combine with the various acidic materials that bring out inflammation, makes its inactivation.3], the lysozyme antivirus action loses by combining with electronegative virus protein, forms double salt with RNA, DNA, apoprotein, makes virally inactivated.4], up-to-date result of study more proves: lysozyme anti-HIV-1 type virus mechanism is the same with RNA enzyme (RNase).And it is reported that coronavirus is a RNA viruses.
Lysozyme is an endogenous active substance, be one of the extraneous self-defense response mechanism of invading of human body self opposing, nonspecific immunity factor, its enhancing immunity effect is by strengthening macrophage and leukocytic phagocytic function, and improve the leukopenia that cytostatics causes, strengthen the immunity of cell.Existing clinical data has proved that it is important step that treatment SARS disease improves autoimmune mechanism.
Lysozyme is a clinical use old medicine for many years, the national standard of existing Chinese medicine, and be the OTC medicine, the lysozyme raw material extracts from egg, and its safety obtains the proof of clinical use for many years.
Mechanism of action and its antiviral characteristics according to lysozyme, we test to lysozyme anti-coronavirus activity with to SARS treatment of diseases and preventive effect, the proof lysozyme has anti-coronavirus effect preferably, the SARS case is had the effect of treatment and prevention.
Summary of the invention
The invention provides new medicinal usage of a kind of lysozyme and disinfectant applications, be used for the SARS prevention and treatment of diseases.
The invention provides the new medicinal usage of a kind of lysozyme, comprise: contain lysozyme and one or more adjuvant or and the drug regimen of excipient, described drug regimen can be prepared to pharmaceutical dosage forms such as injection, transfusion, solution, capsule, tablet, granule, dragee, buccal tablet, enteric coatel tablets, suppository, eye drop, gel, spray.
The invention provides the new disinfectant applications of a kind of lysozyme, comprise: contain lysozyme and one or more adjuvant or and the combination of the disinfectant of excipient, described drug regimen can be prepared to sterilization dosage forms such as handwashing liquid, gel, Emulsion, spray, facial film, disinfectant.
Embodiment
Example one: lysozyme is to the toxic damages test of isolated culture cell
1) test method
Cell culture: passage cell is seeded on the Tissue Culture Plate, puts in 37 ℃ of incubators, grow up to cell monolayer.Measure medicinal liquid: test medicinal liquid: A by following grouping preparation: cell matched group: blank, common culture fluid.B: lysozyme medicinal liquid group, adopt common culture fluid dissolving, through the membrane filtration of 0.22 μ m, make its final concentration reach 200mg/ml.C: lysozyme medicinal liquid group, adopt common culture fluid dissolving, through the membrane filtration of 0.22 μ m, make its final concentration reach 400mg/ml.
Cultured cell is done following processing: change the cell culture fluid in the culture hole into the A-C medicinal liquid, continue to cultivate 72 hours in 37 ℃ of incubators.
Observe lysozyme to the cultured cell degree of impairment: adopt inverted microscope to observe, observe 72h continuously, write down each porocyte situation of change.
2) result of the test
Blank group cell did not have significant change in 72 hours, the cell growth is normal, and cell density increases.200,400mg/ml lysozyme group cell growth has no significant effect.
Example two: the lysozyme antivirus test that exsomatizes
1) test method
Cell culture: passage cell is seeded on the Tissue Culture Plate, puts in 37 ℃ of incubators, grow up to cell monolayer.Measure medicinal liquid: test medicinal liquid: A by following grouping preparation: cell matched group: blank, common culture fluid, also uninfecting virus group.B: infection group and viral infection group, common culture fluid, viral infection, but do not add any test drug.C: positive controls, behind the viral infection, add ribavirin in the common culture fluid, through the membrane filtration of 0.22 μ m, make its final concentration reach 0.25mg/ml.D: the testing drug group, behind the viral infection, adopt common culture fluid dissolving, through the membrane filtration of 0.22 μ m, make its final concentration reach 20mg/ml.The E group: the testing drug group, preparation is with the D group, and through the membrane filtration of 0.22 μ m, its final concentration reaches 80mg/ml, the F group: the testing drug group, preparation is with the D group, and through the membrane filtration of 0.22 μ m, its final concentration reaches 320mg/ml.
Protection test: cultured cell is done following processing: change the cell culture fluid in the culture hole into the A-F medicinal liquid; cultivate after 30 minutes, will add the coronavirus culture solution of certain titre in every hole respectively, 37 ℃ of absorption are after 30 minutes; culture fluid is removed in suction, washes twice with common culture fluid.Add the A-F medicinal liquid again, in 37 ℃ of incubators, continue to cultivate 72 hours.
Therapeutic test: cultured cell is done following processing: change the cell culture fluid in the culture hole into common culture fluid, add the coronavirus culture solution of certain titre in every hole respectively, 37 ℃ of absorption were inhaled the venom of preventing or cure a disease after 30 minutes, washed twice with common culture fluid.Add the A-F medicinal liquid again, in 37 ℃ of incubators, continue to cultivate 72 hours.
Observe the medicine antivirus action: adopt inverted microscope to observe, observe 72h continuously, write down each porocyte pathological changes situation.
2) result of the test
Blank group cell did not have significant change in 72 hours, the cell growth is normal, and cell density increases.Damage appears in the infection group and viral infection group cell, and it shows as cell surface and the graininess projection occurs, broken, dissolving that cell occurs, and last cell death, fragment swims in the culture fluid.Positive controls ribavirin (0.25mg/ml) can effectively suppress the cytopathy that virus causes, its effect shows as the time dilation that the initial cell pathological changes occurs, and the process of cell injury slows down slowly, and the dead chance that occurs of final cell large tracts of land reduces.
Adopting lysozyme to carry out in the protection test, the cytopathy that the 20mg/ml lysozyme can effectively protect cell antagonism virus to cause in early days.Lysozyme (80,320mg/ml) can protect cell effectively to the cytopathy that anti-coronavirus causes, it is characterized by the time that the initial cell pathological changes occurs that delays, the process of the cell injury that slows down reduces the death of cell large tracts of land.
Adopting lysozyme to carry out in the therapeutic test, the cytopathy that the 20mg/ml lysozyme can effectively protect cell antagonism virus to cause in early days.Lysozyme (80,320mg/ml) can protect cell effectively to the cytopathy that anti-coronavirus causes, it is characterized by the time that the initial cell pathological changes occurs that delays, the process of the cell injury that slows down reduces the death of cell large tracts of land.
The stripped antivirus test of the mixed enzyme solution of example three, lysozyme
1) mixed enzyme of employing cylindricality lipase from candida sp and lysozyme makes the lipase activity unit of force reach 5,500 enzyme activity unit/grams (U/g), and lysozyme content is 5, and 300mg/g is made into following test group:
Group | Lipase (U/g) | Lysozyme (mg/g) |
????A | ????5 | ????5 |
????B | ????500 | ????5 |
????C | ????5 | ????300 |
????D | ????500 | ????300 |
2) test method is with example two.
3) result of the test
Blank group cell did not have significant change in 72 hours, the cell growth is normal, and cell density increases.Damage appears in the infection group and viral infection group cell, and it shows not, and the graininess projection appears in cell surface, broken, dissolving that cell occurs, and last cell death, fragment swims in the culture fluid.Positive controls ribavirin (0.25mg/ml) can effectively suppress the cytopathy that virus causes, its effect shows as the time dilation that the initial cell pathological changes occurs, and the process of cell injury slows down slowly, and the dead chance that occurs of final cell large tracts of land reduces.
Carry out in the protection test at employing lysozyme and lipase bulk testing group; the cytopathy that the A group of low dosage proportioning and the D group of high dose proportioning all can effectively protect cell antagonism virus to cause in early days; prolong in time; the protective effect of B, two groups of mixed liquor pair cells of C is better; it is characterized by the time that the initial cell pathological changes occurs that delays; the process of the cell injury that slows down reduces the death of cell large tracts of land.
Adopting lysozyme to carry out in the therapeutic test; the cytopathy that the A group of low dosage proportioning and the D group of high dose proportioning all can effectively protect cell antagonism virus to cause in early days; prolong in time; the protective effect of B, two groups of mixed liquor pair cells of C is better; it is characterized by the time that the initial cell pathological changes occurs that delays; the process of the cell injury that slows down reduces the death of cell large tracts of land.
Example four, lysozyme sheet are to the therapeutical effect of SARS case
Adopt at random, contrast method, in the SARS case, two groups of picked at random, every group 6 example, one group gives lysozyme 100mg/ time, every day 3-6 time, successive administration 10 days is as test group; Organize for lysozyme in contrast, observe the therapeutic effect of lysozyme for other one group patient SARS.
Observed result: compare with matched group, the case heating paresthesia that gives lysozyme medicine group is eased, and body temperature comparatively fast descends and recovers normal, and the course of disease shortens, and the cough symptom is eased even disappears, and chest films showed lamellar shade reduces to some extent, and it is clear that lung marking becomes.
Example five, lysozyme sheet are to SARS high-risk group's preventive effect
Adopt random method, to live in the SARS epidemic-stricken area, daily requirement often appears at the adult of public place, and the men and women does not limit, as SARS high-risk group.Every example gives oral lysozyme enteric coatel tablets, and 50mg/ time, every day 3 times, successive administration 20 days is observed the preventive effect of lysozyme to SARS high-risk group.
Observed result: give lysozyme medicine group and obvious respiratory symptom do not occur, it is normal that body temperature keeps, and chest films showed is normal.
List of references
1、Tenovuo?J.Clinical?application?of?antimicrobial?hostproteins?lacoperoxidase,lysozyme?and?lactoferrin?inxerosotmia:efficacy?and?safety,Oral?Disease,2002,8:23
2、Basschalack?B,et?al,Inactivation?of?gram-negativebacteria?by?lysozyme,Denatured?lysozyme,andlysozyme-derived?peptides?under?high?hydrostatic?pressure,Applied?Environmental?Microbiolgoy,2001,67:339
3、Ibrahim?HR,et?al,A?helix-loop-helix?peptide?at?theupper?lip?of?the?active?site?cleft?of?lysozyme?confers?potentantimicrobial?activity?with?membrane?permeabilizationaction,J?Biol.Chem.2001,276:43767,
4、Lee-Huang,et?al,Lysozyme?and?RNases?as?anti-HIVcomponents?in?β-core?preparations?of?human?chorinoicgonadotropin,PNAS,1999,96:2678;
5、Hill?G,Summary?of?evidence?that?the?foreskin?andlysozyme?may?protest?against?HIV?infection.Data?on?file.
6、Domachowske?JB,Rocombinant?humm?eosinophil-derivednourotoxin/RNase?2?functions?as?an?effective?antiviral?agentagainst?respiratory?syncytial?virus,J.infect.Dis.1998,177:1458,
7、Coronavirus?as?a?possible?cause?of?severe?acuterespiratory?syndrome,Lancet,2003,
8、Marra,MA,et?al,The?genome?sequence?of?theSARS-associated?coronavirus,Sci.2003,108595:1.
Claims (8)
1) a kind of employing lysozyme is used for SARS treatment of diseases and prevention as medicine and disinfectant applications.
2) according to the medicine of claim 1, it is characterized in that containing lysozyme and one or more adjuvant or and the drug regimen of excipient, described drug regimen can be prepared to pharmaceutical dosage forms such as injection, transfusion, solution, capsule, tablet, granule, dragee, buccal tablet, enteric coatel tablets, suppository, eye drop, gel, spray.
3) according to the medicine of claim 2 or and drug regimen, it is characterized in that lysozyme wherein every day dosage at 10mg-10g, be used for SARS treatment of diseases and prevention.
4) according to the disinfectant of claim 1, it is characterized in that containing lysozyme and one or more adjuvant or and the disinfectant combination of excipient, described drug regimen can be prepared to disinfectant such as handwashing liquid, gel, Emulsion, spray, facial film, disinfectant.
5) according to the disinfectant of claim 4, it is characterized in that the lysozyme content in the solution wherein is 0.1mg/ml-1g/ml, the lysozyme content in the various solid preparations is 0.1mg/g-800mg/g.
6) according to the purposes of claim 1, it is characterized in that adopting mixing enzyme preparation be raw material medicine or and disinfectant, wherein contain the lysozyme components in proportions and reach more than 1% of mixing enzyme preparation total amount.
7) according to the purposes of claim 1, it is characterized in that adopting mixing enzyme preparation be raw material medicine or and disinfectant, wherein contain lysozyme composition dosage every day reach the described scope of claim 3 and or the content of lysozyme reach the described scope of claim 5.
8), it is characterized in that adopting lysozyme to be used for other viral infection treatment of diseases and preventions according to the purposes of claim 1.
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CN106075415A (en) * | 2016-06-30 | 2016-11-09 | 史留喜 | A kind of spray treating pharyngolaryngitis and preparation method thereof |
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- 2003-05-20 CN CNA031266010A patent/CN1548152A/en active Pending
Cited By (12)
Publication number | Priority date | Publication date | Assignee | Title |
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CN106075415A (en) * | 2016-06-30 | 2016-11-09 | 史留喜 | A kind of spray treating pharyngolaryngitis and preparation method thereof |
WO2021012789A1 (en) * | 2019-07-19 | 2021-01-28 | 广州新创忆药物临床研究有限公司 | Pharmaceutical composition containing lysozyme and use thereof |
CN112533628A (en) * | 2019-07-19 | 2021-03-19 | 广州新创忆药物临床研究有限公司 | Pharmaceutical composition containing lysozyme and application thereof |
CN112135625A (en) * | 2020-04-27 | 2020-12-25 | 广州新创忆药物临床研究有限公司 | Medicine and food for preventing or treating COVID-19 new coronary pneumonia and application thereof |
CN112135625B (en) * | 2020-04-27 | 2021-07-06 | 广州新创忆药物临床研究有限公司 | Medicine for preventing or treating COVID-19 new coronary pneumonia and application thereof |
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JP2022533876A (en) * | 2020-04-27 | 2022-07-27 | 広州新創憶薬物臨床研究有限公司 | Medicines, foodstuffs and their use to prevent or treat COVID-19 novel coronavirus pneumonia |
JP2023517771A (en) * | 2020-04-27 | 2023-04-26 | 広州新創憶薬物臨床研究有限公司 | Anti-COVID-19 Medicines, Foods, and Uses |
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EP4144363A4 (en) * | 2020-04-27 | 2024-05-29 | Guangzhou Century Clinical Research Co., Ltd | Drug, food and application of anti-coronavirus infection |
JP7543427B2 (en) | 2020-04-27 | 2024-09-02 | 広州新創憶薬物臨床研究有限公司 | Medicines, foods and uses for anti-COVID-19 infection |
WO2022041664A1 (en) * | 2020-08-25 | 2022-03-03 | 杭州国光药业股份有限公司 | Color modifier, lysozyme lozenge, and method for preparing lysozyme lozenge |
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