CN103191183A - Use of palchouli oil in preparation of antiviral drug - Google Patents
Use of palchouli oil in preparation of antiviral drug Download PDFInfo
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Abstract
The invention discloses use of palchouli oil in preparation of an antiviral drug, and discloses an antiviral pharmaceutical composition, which is a drug prepared from the palchouli oil as an active ingredient and a pharmaceutically acceptable auxiliary material or an auxiliary ingredient. The palchouli oil disclosed by the invention is significant in anti-virus effect, can treat a disease caused by virus, is accurate in curative effect on vital myocarditis and pneumonia, and has strong practical application value.
Description
Technical field
The present invention relates to the new purposes of oil of Herba Pogostemonis, particularly the purposes of oil of Herba Pogostemonis in the preparation antiviral drugs.
Background technology
By the infectious disease that virus causes, serious threat always human life with healthy.According to statistics, the infectious disease more than 80% is to be caused by virus.Viral disease has related to clinical medical every field, and disease viral and that cause becomes the focus that many subjects are paid close attention at present, and modal in the virus infection is exactly by catching that Respirovirus and enterovirus are caused.
Respirovirus is that a big class can be invaded respiratory tract and causes the respiratory tract local patholoic change, or is the virus that portal of entry causes the respiratory tissues organ lesion with the respiratory tract only, is the main pathogens that causes the acute respiration road transmission.At present, the known virus of acute respiratory infection that can cause has kind more than 10, comprises influenza virus, parainfluenza virus, adenovirus, herpes simplex virus, respiratory syncytial virus etc.According to statistics, 90% above acute respiratory infection causes by virus, is clinically commonly encountered diseases and frequently-occurring disease with upper respiratory tract infection especially, and severe patient is bronchiolitis and pneumonia among the infected, can cause acute respiratory distress and heart failure, even causes death.Respirovirus easily morphs, and it is pathogenic thereby increase, or causes some new diseases.Wherein influenza virus (influenza) is the main Causative virus that causes acute respiratory infection disease influenza, influenza have infectiousness strong, propagate fast, crowd characteristics such as susceptible repeatedly.Issue up-to-date epidemic situation circular according to World Health Organization (WHO), only this year so far first type HINI influenza oneself causes in the whole world and surpasses 10,000 people's death, its in the Central Asia, the activity of South Asia region continues to strengthen especially.China is the multiple district of influenza, and popular or local the breaking out basically of influenza all can be taken place every year, makes people more than 100,000,000 suffer the puzzlement of influenza, and because influenza surpasses 500,000 people to the hospitalier of hospital, the society that causes and financial burden are difficult to statistics.The harm of adenovirus also can not be ignored, adenovirus AdV(Adneovirus) is the common virus of childrens respiratory tract infection, the most normal disease that causes is pharyngoamygdalitis, secondly be pneumonia, gastroenteritis, bronchitis and otitis media, also can cause pharynx-conjunctivitis, encephalitis, keratitis, enteritis etc., case fatality rate is very high.
Coxsackie virus (Coxsaekieviurs) belongs to Picornaviridae (Picomaviridea), and enterovirus genus (Enterroviurs) can be divided into A group and B group according to its difference to the neonatal rat pathogenecity.Coxsackie B virus (Coxsackievirusgroup B, CVB) can cause epidemic pleurodynia, aseptic encephalitis, meningoencephalitis and pericarditis, especially CVB3 is the main pathogens of viral myocarditis, can cause that focal necrosis takes place in cardiac muscular tissue, and with inflammatory cell infiltration, pathological changes such as myocardial cell cracking, persistent viral infection can cause DCM (dilated cardiomyopathy), has brought great threat to the mankind.
Since the nineties in 20th century, research worker is found many new antiviral drugs, kind surplus the antiviral drugs of using has clinically surpassed 20 at present, but its definite curative effect still has very big dispute, and after the antiviral chemosynthesis class medicine prolonged application, virus easily produces drug resistance to it, has now become the thorny problem for the treatment of viral disease clinically.Routine is used for preventing and treating medicine for treating viral infections virazole (ribavirin) now, by inhibition phosphoric acid time fast nuclear dehydrogenase, thereby suppresses virus replication, and medication in early days is effective.But there is medullary cell toxicity in the virazole vein when giving, therefore preferably only give through respiratory tract by a spot of aerosol, but leukopenia often appears in the clinical practice, can cause side effect such as headache, irreversible anemia, serum bilirubin rising when dosage is excessive, zoopery has the report of teratogenesis.Therefore, the research of antiviral drugs has important and practical meanings, and the research of antiviral drugs of seeking a kind of efficient, safety, few side effects is imperative.
The natural resources of Chinese medicinal materials of China is very abundant, and applicating history is long, has accumulated a lot of data and experience in long-term practice.The Chinese medicine curative effect affirms that toxic and side effects is little, and the medicine source is abundant and cheap.Propose to have heat clearing away in the theory of Chinese medical science, Detoxication, as single medicines such as Flos Lonicerae, Folium Isatidis, Rhizoma Atractylodis, the Radixs Astragali, and compound medicine such as Radix Isatidis electuary can be made the most of the advantage at anti-virus aspect.Experiment in vitro and clinical practice prove, most viruses can obtain better curative effect to the Chinese herbal medicine sensitivity as Respirovirus (influenza virus, Measles virus, mumps virus), enterovirus (the scorching virus of ridge ash, rotavirus, Coxsackie virus), the use treatments by Chinese herbs such as (epidemic encephalitis B virus, encephalitises) of sooty mould poison.The general pharmacological action principle is by suppressing virus replication, stop virus to cause the pathological changes of cell, regulate immunization, improve pulmonary circulation, comprehensive functions such as minimizing and the pathogenic position of elimination inflammatory exudate, make disease of viral infection sx or recovery from illness, so the Chinese medicine antiviral there are potential advantages and wide prospect.
Herba Pogostemonis is the perennial thorn stamen of Labiatae (Labiatae) herbaceous plant Herba Pogostemonis (Pogostemon cablin (Blanco) Benth.), it is fragrant to have another name called branch, originate in Tropical Asia such as Philippine, existing China, India, Japan, Indonesia, Malaysia, Madagascar, Brazil, Paraguay and the Russia etc. of extensively being distributed in.At present wide flower bud perfume (or spice) in Guangzhou, Zhaoqing, Zhanjiang and the Hainan in Guangdong, all there are cultivation in provinces (district) such as Guangxi, Sichuan, be one of " ten Da Nan medicines ", its medical material commodity are divided into board perfume (or spice) (Guangzhou product), branch fragrant (Zhaoqing product), fragrant (Hainan product) 4 kinds of profound fragrant (Zhanjiang product) and south by place of production difference.Its acrid in the mouth, slightly warm in nature is returned spleen, stomach, lung meridian, with all herbal medicine, has eliminating turbid pathogen with aromatics, the preventing or arresting vomiting that whets the appetite, delivers the effect of expelling summer-heat.Be usually used in the vomiting of turbid damp obstructing in middle-JIAO, gastral cavity painful abdominal mass, the heat-damp in summer asthenia, uncomfortable in chestly do not relax, cold-damp is closed summer-heat, stomachache vomiting and diarrhoea, nasosinusitis headache, affection of exogenous wind-cold.The volatile oil of Herba Pogostemonis has antiinflammatory, analgesia and antibacterial activity.
Herba Pogostemonis is the dry aerial parts of labiate Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze., contains volatile oil, the same Herba Pogostemonis of effect.Modern pharmacology shows, Herba Pogostemonis has effects such as resisting pathogenic microbes, antiinflammatory, analgesic, analgesia, be the main component for the treatment of such as HUOXIANG ZHENGQI SHUI, antivirus oral liquid influenzas, flu Chinese patent medicine commonly used, but its effective substance is still indeterminate and do not see that resisiting influenza virus effect report in its body is arranged.The volatile oil of Herba Pogostemonis has antibacterial action.
Summary of the invention
The object of the present invention is to provide the new purposes of oil of Herba Pogostemonis, and be the pharmaceutical composition of active component with the oil of Herba Pogostemonis.
The new purposes of oil of Herba Pogostemonis of the present invention is the purposes in the preparation antiviral drugs.
Described antiviral drugs is the medicine of resisiting influenza virus, CVB-3 and/or adenovirus.
Described antiviral drugs is that treatment respiratory tract infectious disease, pneumonia, otitis media, viral myocarditis, conjunctivitis, encephalitis, keratitis, hepatitis are or/and the medicine of enteritis.Wherein, the medicine of described treatment respiratory tract infectious disease is that treatment rhinitis, pharyngitis, tonsillitis, laryngitis, tracheitis are or/and bronchitic medicine.
Wherein, described oil of Herba Pogostemonis derives from the volatile oil of Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze. extraction.
Wherein, the described oil of Herba Pogostemonis that comes from Herba Pogostemonis Pogostemon cablin (Blanco) Benth., patchouli alcohol content is not less than 40%(w/w), Pogostone content is not less than 20%(w/w).
Described oil of Herba Pogostemonis is prepared by following method: the herb of getting Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze., be ground into coarse powder, add water, soak, the employing steam distillation extracts, and namely gets oil of Herba Pogostemonis.
Antiviral medicinal composition of the present invention is to be active component with the oil of Herba Pogostemonis, adds the medicament that acceptable accessories or complementary composition are prepared from.
Described medicament is external preparation, oral formulations or ejection preparation.
Described external preparation is lotion, suppository, liniment or liniment.
The content of oil of Herba Pogostemonis is 0.1%～100%(w/w) in the described preparation.
Oil of Herba Pogostemonis of the present invention has antiviral effect, especially the inhibitory action to influenza virus, CVB-3 and/or adenovirus is remarkable, acute respiratory infection and other inflammation that virus is caused have therapeutical effect, wherein, to the determined curative effect of viral myocarditis, pneumonia, has good potential applicability in clinical practice.
Obviously, according to foregoing of the present invention, according to ordinary skill knowledge and the customary means of this area, not breaking away under the above-mentioned basic fundamental thought of the present invention prerequisite, can also make modification, replacement or the change of other various ways.
The specific embodiment of form is described in further detail foregoing of the present invention again by the following examples.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.
The specific embodiment
The preparation of embodiment 1 oil of Herba Pogostemonis of the present invention
Get Herba Pogostemonis Pogostemon cablin (Blanco) Benth. herb, pulverized the 20-40 mesh sieve, add the distilled water of 10-14 times of weight, soak after 1-5 hour, adopt steam distillation to extract 2-6 hour, namely get oil of Herba Pogostemonis of the present invention, claim patchouli oil again, wherein, contain patchouli alcohol (C
15H
26O) must not be lower than 40%w/w, Pogostone must not be lower than 20%w/w.
The preparation of embodiment 2 oil of Herba Pogostemonis of the present invention
Get Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze. herb, pulverized the 20-40 mesh sieve, add the distilled water of 10-14 times of weight, soak after 1-5 hour, adopt steam distillation to extract 2-6 hour, namely get oil of Herba Pogostemonis of the present invention.
Oil of Herba Pogostemonis of the present invention also can extract according to pharmacopeia (version in 2005) appendix XD determination of volatile oil method, also can adopt organic solvent extraction, supercritical CO
2Prior aries such as extraction are extracted, and perhaps obtain by buying the commercially available prod.
Below prove beneficial effect of the present invention by the test of pesticide effectiveness:
Experimental example 1 oil of Herba Pogostemonis extracorporeal antivirus effect experiment of the present invention
1, material and instrument
1.1 cell strain and Strain
1.1.1 cell strain:
Mdck cell (Madin-Darby canine kidney(cell line)), Hep-2 cell (people's laryngeal carcinoma epithelial cell), HeLa cell (cervical cancer cell) are available from Chinese medicine and biological products assay institute.
1.1.2 Strain:
Influenza A virus is selected the reassortant virus strain for use, and serotype is that H1N1(is hereinafter to be referred as influenza virus), be derived from national influenza center; Coxsackie virus selects for use B to organize 3 types (hereinafter to be referred as CVB-3), is derived from visiting center, Sichuan Province; Adenovirus is selected common 3 types (Ad-3) for use, is presented by Gansu Province Disease Control and Prevention Center.
1.2 medicine and control drug
Trial drug: the patchouli oil of invention medicine: embodiment 1 preparation.
Positive drug: ribavirin injection: specification is 100mg/ml, and Tianjin Pharmaceutical Jiaozuo Co., Ltd. produces.Authentication code: the accurate word H19992467 of traditional Chinese medicines, product batch number: 10110521; The injection acyclovir: specification is the 0.25g/ bottle, and Tianjin Pharmaceutical Jiaozuo Co., Ltd. produces.Authentication code: the accurate word H20034034 of traditional Chinese medicines, product batch number: 10122403.
1.3 culture medium, reagent and consumptive material
RPMI-1640 culture medium (Gibco
TMCompany, lot number 1313945); DMEM culture medium (Gibco
TMCompany, lot number 1345538); Hyclone (HyClone biochemistry goods (Beijing) company limited, lot number NVM0347); Trypsin Amresco company); Penicillin G sodium, the aseptic Tissue Culture Flask of streptomycin and 96 porocyte culture plates (U.S. Corning company).1,5, the 10ml Dispette, the disposable rifle head of 200 μ l, 1ml (the biological consumptive material in Haimen, Jiangsu company)
1.4 key instrument
Water isolation type constant temperature CO
2Incubator (model MCO-15AC, SANYO GS company);
OLYMPUS inverted microscope (model C KX41, Japanese Olympus company);
Micropipettor (French GILSON company produce);
Water isolation type electro-heating standing-temperature cultivator (model GSV-DA-1, Huangshi, Hubei Province medical apparatus and instruments factory);
Horizontal centrifuge (model LXJ-II, Shanghai medical analytical instrument factory);
Electronic balance (model JA-2603, Shanghai balance equipment factory).
2, test method
2.1 the pre-treatment of medicine and positive control
Elder generation is the white liquid preparation with 0.5% tween 80 hydrotropy with invention medicine patchouli oil, and its initial concentration is that 20%(patchouli oil density is 1.012g/ml, concentration 202.4mg/ml).With 0.5% tween 80 this white liquid is done 1/10 dilution before using, standby after the filtration sterilization.
To do 1/10 dilution (being 10mg/ml) with standby with sterilized water for the positive control drug 'Libaweilin ' injection of influenza virus and Coxsackie virus with for the positive control medicine injection acyclovir of adenovirus.
The mensuration of 2.250% histiocyte infective dose
Influenza virus and adenovirus are diluted to the viral liquid of variable concentrations with 3% hyclone DMEM, Coxsackie B virus 3 usefulness 3% hyclone RPMI-1640.
Influenza virus inoculation mdck cell, Coxsackie B virus 3 strains inoculation HeLa cell, adenovirus inoculation Hep-2 cell, in 37 ℃, 5%CO
2The observation infection cell is justified cytopathy (cytopathiceffect such as contracting, come off after cultivating 7d in the incubator, CPE) situation, with measure the infective dose that each virus causes 50% pathogenic histiocyte (50%tissue culture infective dose, TCID50).
2.3 antiviral experiment
In the MDCK of 96 well culture plates cell monolayer, add 100 * TCID50 influenza virus liquid 0.1ml, in the HeLa cell monolayer, add 100 * TCID50CVB-3 virus liquid 0.1ml and in the Hep-2 cell monolayer, add 100 * TCID50Ad-3 virus liquid 0.1ml, in 37 ℃, 5%CO
2Adsorbed 2 hours, and inhaled and abandon viral liquid, add dilution good invention medicine and positive control drug respectively, each concentration is done 3 multiple holes.Experiment arranges the infection contrast of not administration of infection equivalent virus and the normal cell contrast of virus-free infection simultaneously.Each culture plate is in 37 ℃, 5%CO
2Continue to cultivate 5d, every day, microscopically was observed CPE.
3. experimental result and pharmacodynamic index calculate
3.1 the TCID that each is viral
50Value
CPE occurs in about 3 days behind the influenza virus inoculation mdck cell, occurred CPE behind Coxsackie B virus 3 inoculation HeLa cells and the adenovirus Ad-3 inoculation Hep-2 cell in about 5 days.It is 10 that the TCID50 of influenza virus tires
-4, the TCID50 of Coxsackie B virus 3 and adenovirus Ad-3 tires and is 10
-12
3.3 invention medicine antiviral experiment effect and pharmacodynamic index calculate
3.3.1 experimental result
Experimental result is as shown in table 1:
Table 1 invention medicine antiviral experimental result (n=3)
As can be seen from Table 1, medicine of the present invention is good to the inhibition of influenza virus, Coxsackie B virus 3 strains and adenovirus.Reach identical viral suppression ratio (75%), amount of drug of the present invention is 0.092mg/ml, and the consumption of positive drug is 0.156mg/ml, is 1.7 times of amount of drug of the present invention.
Description of test, the effect that oil of Herba Pogostemonis of the present invention suppresses influenza A virus, CVB-3 and adenovirus is good.
Experimental example 2 oil of Herba Pogostemonis interior resisting virus experiments of the present invention
1, material and instrument
1.1 Strain
Influenza A virus is selected the reassortant virus strain for use, and serotype is that H1N1(is hereinafter to be referred as influenza virus), be derived from national influenza center; Coxsackie virus selects for use B to organize 3 types (hereinafter to be referred as CVB-3), is derived from visiting center, Sichuan Province; Adenovirus is selected common 3 types (Ad-3) for use, is presented by Gansu Province Disease Control and Prevention Center.
1.2 medicine and control drug
Trial drug: the patchouli oil of invention medicine: embodiment 1 preparation.
Positive drug: ribavirin injection: specification is 100mg/ml, and Tianjin Pharmaceutical Jiaozuo Co., Ltd. produces.Authentication code: the accurate word H19992467 of traditional Chinese medicines, product batch number: 10110521; The injection acyclovir: specification is the 0.25g/ bottle, and Tianjin Pharmaceutical Jiaozuo Co., Ltd. produces.Authentication code: the accurate word H20034034 of traditional Chinese medicines, product batch number: 10122403.
1.3 animal: mice Bab/c level hero, body weight 10 scholar 2g, male and female half and half
Mice Bab/c level hero, body weight 18 scholar 2g, male and female half and half
1.4 culture medium, reagent and consumptive material
RPMI-1640 culture medium (Gibco
TMCompany, lot number 1313945); DMEM culture medium (Gibco
TMCompany, lot number 1345538); Hyclone (HyClone biochemistry goods (Beijing) company limited, lot number NVM0347); Trypsin Amresco company); Penicillin G sodium, the aseptic Tissue Culture Flask of streptomycin and 96 porocyte culture plates (U.S. Corning company).1,5, the 10ml Dispette, the disposable rifle head of 200 μ l, 1ml (the biological consumptive material in Haimen, Jiangsu company)
Ether (analytical pure, the Long Huagongshijichang of Chengdu section, lot number: 20111215); 0.9% normal saline (Cologne, Chengdu pharmaceutical Co. Ltd, lot number: H120115)
Lactic acid dehydrogenase (LDH) is measured test kit, and bio-engineering research institute, lot number: 20121013 are built up in Nanjing; Superoxide dismutase (SOD) is measured test kit, and bio-engineering research institute, lot number: 20121012 are built up in Nanjing; Malonaldehyde (MDA) is measured test kit, and bio-engineering research institute, lot number: 20121010 are built up in Nanjing; The CK-MB(creatine kinase isozyme), TNF-α (tumor-activated factor-alpha) measures test kit, U.S. RD company, lot number: 201210
1.4 key instrument
Water isolation type constant temperature CO
2Incubator (model MCO-15AC, SANYO GS company);
OLYMPUS inverted microscope (model C KX41, Japanese Olympus company);
Micropipettor (French GILSON company produce);
Water isolation type electro-heating standing-temperature cultivator (model GSV-DA-1, Huangshi, Hubei Province medical apparatus and instruments factory);
Horizontal centrifuge (model LXJ-II, Shanghai medical analytical instrument factory);
Electronic balance (model JA-2603, Shanghai balance equipment factory);
Microplate reader, U.S. Thermo Scientific company, model: Varioskan Flash.
2, test method
2.1 the pre-treatment of medicine and positive control
Elder generation is the white liquid preparation with 0.5% tween 80 hydrotropy with invention medicine oil of Herba Pogostemonis, and its initial concentration is that 20%(oil of Herba Pogostemonis density is 1.012g/ml, concentration 202.4mg/ml).With 0.5% tween 80 this white liquid is done 1/10 dilution before using, standby after the filtration sterilization.
To do 1/10 dilution (being 10mg/ml) with standby with sterilized water for the positive control drug 'Libaweilin ' injection of influenza virus and Coxsackie virus with for the positive control medicine injection acyclovir of adenovirus.
2.2 invention medicine interior resisting virus experiment
The foundation of model 2.2.1 the viral infection animal is caused a disease
Get 40 of Bab/c mices, the SPF level, body weight 10 scholar 2g are divided into 4 groups at random, 10 every group.Behind the injection site routine disinfection, by group respectively former times of lumbar injection, 1/2 and 1/4 CVB3 virus liquid 0.3ml, the normal control group adopts 0.3ml normal saline lumbar injection.
Other gets the Bab/c mice, SPF level, body weight 18 scholar 2g, random packet, 10 every group.Slightly anaesthetize with ether earlier by group, then respectively the collunarium infector doubly, 1/2 and 1/4 influenza virus liquid or adenopathy venom 1ml, the normal control group adopts 1ml normal saline collunarium.
Animal housing keeps air fresh, relative humidity 60%, 2 ℃ of temperature 22 scholars, every day is supplement feed, moisture, replacing bedding and padding regularly, observe morbidity and the death condition of mice, and record it, to determine whether successfully to set up the pathogenic model of CVB3, influenza virus and adenovirus infection animal.
The model test 2.2.2 invention medicine interior resisting virus causes a disease
2.2.2.1 invention medicine interior resisting virus myocarditis model test
Get 140 of Bab/c mices, body weight 10 scholar 2g are divided into 7 groups at random, 20 every group.Make marks.
With patchouli oil clinical injection consumption and LD
50Be foundation, in the safe dose scope, set up the medication therapy groups of high, medium and low, ultralow 4 kinds of concentration respectively, i.e. the 1.25%(0.127g/kg of medicine original concentration), 0.95%(0.096g/kg), 0.625%(0.063g/kg) and 0.48%(0.049g/kg).Normal control group, CVB3 virus model matched group and ribavirin positive drug matched group (positive drug concentration is 0.15g/kg) are set simultaneously.
With injected in mice position routine disinfection, except the normal control group, other respectively organize the required CVB3 virus liquid of lumbar injection virus model, and the normal control group adopts the normal saline lumbar injection, and injection volume is identical with other papova liquid injection volumes.Begin intraperitoneal injection behind the 24h, injection volume is 0.1ml/10g, every day 1 time.Animal housing keeps air fresh, relative humidity 60%, 2 ℃ of temperature 22 scholars, every day is supplement feed, moisture, replacing bedding and padding regularly, treat 7 days continuously after, stop administration and observe the 15th day.Observe morbidity sign and the death condition of mice every day, take by weighing body weight and record.
Started at the same day by the treatment administration, and put to death 4 at random from each treated animal respectively when the 5th day, the 10th day and the 15th day, eye socket is got blood and is collected, and takes out heart, liver, lung, kidney, remove surperficial blood with filter paper after, conscience lung kidney body rate is calculated in weighing.Then each internal organs is got 100mg, with the abundant homogenate of RPMI-1640 cell maintenance medium 2ml.After tissue homogenate is used the filtering with microporous membrane degerming, multigelation three times, the centrifugal 30min of 3000rPm gets supernatant 0.1ml and is inoculated in 96 orifice plates that grow into monolayer Hela cell, and each sample is done 3 multiple holes, 37 ℃, 5%CO2 discards liquid in the hole after hatching 2h, and PBS adds 1640 cell maintenance mediums after washing 3 times, 37 ℃, 5%C02 cultivates 72h-120h.Every day observation of cell growing state and corresponding CPE degree, CPE person not occurring does not still have CPE for 3 times then for the virus-free infection person through blind passage.Eye socket is got the blood that blood collects spend the night in 4 ℃ of placements, the centrifugal 20min of 3000rpm gets serum and carries out the serum biochemistry index and detect.
2.2.2.2 anti-pneumonia model test in the invention medicine body
Get 140 of Bab/c mices, body weight 18 scholar 2g, random packet, 20 every group.Make marks.
Be foundation with the clinical gastric infusion consumption of patchouli oil, in the safe dose scope, set up the medication therapy groups of high, medium and low, ultralow 4 kinds of concentration respectively, i.e. the 10%(1.27g/kg of medicine original concentration), 5%(0.508g/kg), 2.5%(0.254g/kg) and 1.25%(0.127g/kg).Normal control group, influenza virus model control group, adenovirus model control group, ribavirin positive drug matched group and acyclovir positive drug matched group are set simultaneously.
Except the normal control group, model group infects for respectively the mice collunarium by modeling success required influenza virus and adenovirus virus quantity, and the normal control group adopts the normal saline collunarium, and the collunarium amount is identical with other papova liquid infective doses.Begin gastric infusion behind the 24h, irritating the stomach amount is 0.3ml/10g, every day 1 time.
Animal housing keeps air fresh, relative humidity 60%, 2 ℃ of temperature 22 scholars, every day is supplement feed, moisture, replacing bedding and padding regularly, treat 7 days continuously after, stop administration and observe the 15th day.Observe morbidity sign and the death condition of mice every day, take by weighing body weight and record.
Started at the same day by the treatment administration, when the 5th day, the 10th day and the 15th day, put to death 4 dissections at random from each treated animal respectively, perusal phase pulmonary lesion situation, after winning full lung and removing surperficial blood with filter paper, its lung body rate is calculated in weighing.Grind the Mus lung then and inoculate corresponding sensitive cells, to determine that whether the pneumonopathy change is by due to this test viral infection.
3, result of the test
3.1 mice body inner virus model is set up
With former times, 1/2 and 1/4 CVB3 virus liquid intraperitoneal injection of mice, 0.3ml/ only observes and finds that mice hair before injecting virus is glossy, and activity, diet, defecation are normal.Behind the injectable drug, compare with normal control group mice, the minimizing that came into play at the 2nd～3 day of former times of viral liquid group mice, dietary amount descends, and loses weight, hair tarnishes, and frizzle appears, and died off in the 6th～7 day, all dead during by the 8th day, dissect and find that heart is obviously little than normal group mice, simultaneously visible milky point-like, streak class fat-like pathological changes under the heart adventitia.1/2 former times of concentration group and 1/4 former times of concentration group mice also appearance activity minimizing in the time of the 2nd～3 day, dietary amount descends, and loses weight, but began the increase activity in the time of the 5th day, and dietary amount increases, and state returns to normal group mice state gradually.Illustrate set up the viral myocarditis model need be with former times of CVB3 virus liquid intraperitoneal injection of mice, 0.3ml/ is only.
With former times, 1/2 and 1/4 influenza virus liquid, adenopathy venom collunarium infecting mouse respectively, 1ml/ only, observe to find that model mice compares with normal control group mice, former times of viral liquid group mice began to occur successively death at collunarium the same day, and be all dead in about the 4th day; 1/2 former times of concentration group reduces appearance activity in the 2nd～3 day, and dietary amount descends, and hair tarnishes, and frizzle occurs, occurs death in succession on the 5th～6 day, and is all dead in the time of the 7th day, dissects and finds that pulmonary lesion is obvious.Malaise symptoms appearred in 1/4 former times of concentration group at the 2nd day, but took a turn for the better in succession in the 3rd～4 day state.Illustrate that influenza virus and the adenovirus of setting up pneumonia model needs are 1/2 former times of concentration group, 1ml/ only.
3.2 invention medicine interior resisting virus effect
3.2.1 the invention medicine is to the effect of mice with viral myocarditis model
3.2.1.1 ordinary circumstance influence
Experimental result is as shown in table 2:
Table 2 each experimental mice body weight and death condition
| Group | 0d | 5d | 10d | 14d | Total death toll |
| Normal group | 12.0±0.5 | 13.3±0.5 | 15.0±0.5 | 18.9±2.1 | 0/20 |
| Model group | 12.5±0.4 | 10.3±0.5 | 12.0±1.5 | 17.5±0.5 | 18/20 |
| Positive group (0.15g/kg) | 12.0±0.5 | 11.8±0.7 | 12.6±1.2 | 15.3±1.0 | 8/20 |
| Invention medicine 1.25% (0.127g/kg) | 11.5±0.6 | 9.8±0.7 | 15.0±1.5 | 16.8±1.5 | 14/20 |
| Invention medicine 0.95% (0.096g/kg) | 12.0±0.5 | 9.9±0.2 | 10.8±0.7 | 17.2±1.6 | 6/20 |
| Invention medicine 0.63% (0.063g/kg) | 11.8±0.5 | 10.5±0.5 | 12.1±1.2 | 14.5±1.2 | 8/20 |
| Invention medicine 0.48%(0.049g/kg) | 12.0±0.5 | 10.5±0.5 | 11.5±1.0 | 13.8±1.2 | 13/20 |
The action of normal group mice is normal, and when finishing to observation, mortality rate is 0%;
Model group, positive group and invention medicine group mice all occur losing weight when modeling finishes in various degree, and companion's fur is not damp, movable, the minimizing of ingesting, the increase of urinating;
The model group mice began to die off in the 6th～7 day, observed when finishing, and mortality rate is up to 90%;
The symptom of positive drug group and invention medicine group decimal all alleviates, and activity, food ration increase, and body weight begins to rise:
Wherein, the mortality rate of positive group is 40%; The mortality rate of invention medicine 1.25% is 70%; The mortality rate of invention medicine 1.25% is 30%; The mortality rate of invention medicine 0.63% is 40%; The mortality rate of invention medicine 0.48% is 65%.
Oil of Herba Pogostemonis of the present invention raises earlier to the increase along with consumption of the therapeutic effect of viral myocarditis, and the back reduces, and is the effect optimum of 0.096g/kg with the consumption.
When patchouli oil using dosage of the present invention was 0.063～0.096g/kg, therapeutic effect was better than positive drug (0.15kg/g), illustrated that medicine of the present invention is good to the therapeutic effect of viral myocarditis.
3.2.1.2 the invention medicine is to the influence of mice cardiopulmonary kidney liver body rate
Experimental result is as shown in table 3:
Body weight and the cardiopulmonary kidney liver body rate result (n=4) of each experimental mice of table 3 different sample times
| Time | Project | Normal group | Model group | Positive group | 1.250% | 0.950% | 0.625% | 0.480% |
| 5d | Body weight | 12.3±0.5 | 10.3±0.5 | 11.8±0.7 | 9.8±0.7 | 9.9±0.2 | 10.5±0.5 | 10.5±0.5 |
| ? | Heart body rate | 0.6315±0.0235 | 0.6597±0.0663 | 0.7184±0.0042 | 0.5996±0.0046 | 0.6821±0.0112 | 0.6076±0.0848 | 0.6419±0.0109 |
| ? | Lung body rate | 0.8166±0.0450 | 0.8887±0.0783 | 0.8332±0.0399 | 0.9207±0.0077 | 0.7389±0.0453 | 0.9248±0.0257 | 0.8686±0.0932 |
| ? | Kidney body rate | 1.3447±0.0286 | 1.6533±0.0867 | 1.5537±0.102 | 1.4497±0.0234 | 1.1958±0.0611 | 1.5233±0.1719 | 1.5800±0.0665 |
| ? | Liver body rate | 4.4300±0.3908 | 3.7796±0.3976 | 4.1572±0.4694 | 4.4444±0.5096 | 5.0274±0.9181 | 3.8457±0.2019 | 5.2962±1.0982 |
| 10d | Body weight | 15.0±0.5 | 12.0±1.5 | 12.6±1.2 | 15.0±1.5 | 10.8±0.7 | 12.1±1.2 | 11.5±1.0 |
| ? | Heart body rate | 0.5877±0.0601 | 0.5232±0.0546 | 0.5922±0.0922 | 0.5253±0.0192 | 0.5843±0.0277 | 0.5388±0.0812 | 0.5606±0.0564 |
| ? | Lung body rate | 0.7314±0.0653 | 0.7073±0.1145 | 0.7642±0.0773 | 0.7402±0.0998 | 0.7428±0.0332 | 0.8637±0.1201 | 0.7274±0.0124 |
| ? | Kidney body rate | 1.3330±0.0909 | 1.2767±0.2100 | 1.4587±0.1702 | 1.2835±0.1712 | 1.3658±0.0299 | 1.4435±0.9087 | 1.2995±0.1021 |
| ? | Liver body rate | 5.5674±0.9655 | 4.7726±1.3837 | 4.4343±0.9207 | 4.8972±0.2366 | 5.1760±0.8779 | 3.8525±2.0364 | 4.3800±0.34 |
| 14d | Body weight | 18.9±2.1 | 17.5±2.0 | 15.3±1.0 | 16.8±1.5 | 17.2±1.6 | 14.5±1.2 | 12.8±1.2 |
| ? | Heart body rate | 0.4601±0.0312 | 0.4960±0.0213 | 0.4881±0.0505 | 0.4555±0.0368 | 0.4807±0.0093 | 0.5966±0.0557 | 0.4641±0.0634 |
| ? | Lung body rate | 0.6754±0.0846 | 0.6954±0.264 | 0.7439±0.0892 | 0.8072±0.0385 | 0.6291±0.0150 | 0.7214±0.0854 | 0.7187±0.1611 |
| ? | Kidney body rate | 1.0997±0.0570 | 1.1669±0.0690 | 1.3179±0.1507 | 1.2525±0.0993 | 1.3243±0.0551 | 1.2476±0.0229 | 1.1804±0.1223 |
| ? | Liver body rate | 4.3529±0.2331 | 4.4303±0.8590 | 4.5229±1.2381 | 4.1260±0.8362 | 5.5778±0.0487 | 5.0745±0.5338 | 4.0730±1.0212 |
Normal group mice cardiopulmonary kidney liver device is red and gloss all;
Model group heart color is dim and dwindle and simultaneously milky point-like, streak class fat-like occur with visceral pericardium, the lungs enlargement, and Liver and kidney does not show and changes;
The heart of positive group and each concentration group of invention medicine the dim color that narrows down to of color occurs and recovers gradually, the trend that visceral pericardium milky point-like, streak class fat-like reduce, and lungs enlargement degree alleviates.
Patchouli oil of the present invention can be alleviated every organ disease, and the color that can effectively dwindle heart is dim to recovering normal gradually, further illustrates patchouli oil of the present invention and can alleviate viral myocarditis.
3.2.1.3 the invention medicine is to the influence of mice serum biochemical indicator
The mice serum of virus inoculation 5d, 10d and 14d is done the serum biochemistry index detect, normal group is compared with model group, and LDH, CK-MB and TNF-alpha levels and MDA content are significantly low, SOD active obviously high (P＜0.01); Positive group is compared with model group with each concentration group of invention medicine, LDH, CK-MB and TNF-alpha levels and MDA content have reduction in various degree, the SOD activity then increases (P＜0.01) in various degree, and positive group and each concentration group of invention medicine are compared LDH, CK-MB and TNF-alpha levels and MDA content in various degree increase are arranged with normal group, and the SOD activity then reduces (P＜0.05) in various degree.The LDH of normal group and CK-MB level are along with the increase of time changes not quite, and the SOD activity has less increase, and MDA content and TNF-alpha levels then obviously reduce; The LDH of model group, CK-MB level are along with the increase of time obviously reduces, and SOD activity, MDA content and TNF-alpha levels then significantly increase; The LDH of positive group, 0.95% and 0.625% concentration group and CK-MB level are along with the less reduction of the increase of time, and SOD is active to be increased, and MDA content and TNF-alpha levels then obviously reduce, and the index variation all is tending towards the normal group level; The LDH of 1.25% and 0.48% concentration group, CK-MB level, MDA content and TNF-alpha levels are along with the increase of time reduces, and SOD is active to be increased, and the index variation is tending towards the model group level.Each experimental mice serum biochemistry index the results are shown in Table 4.
The serum biochemistry index (n=4) of each experimental mice of table 4 different sample times
(
*Compare with model group, P＜0.01,
#Compare P＜0.05 with normal group)
As can be seen from Table 4, the invention medicine can significantly reduce LDH, CK-MB and TNF-alpha levels and MDA content, improves the SOD activity, compares with model group, significant difference (P＜0.01) has verified that further patchouli oil of the present invention is to the therapeutical effect of viral myocarditis.
3.2.2 the invention medicine is to the effect of pneumonia mouse model
3.2.2.1 ordinary circumstance influence
After viral collunarium infected 2～3 days, losing weight all appearred in virus group mice in various degree, and companion's fur is not damp, movable, the minimizing of ingesting, the increase of urinating; The model group mice began to die off in the 6th～7 day, only remained 2 after 12 days until observing end; 4 concentration groups of invention medicine mice loses weight obvious than model group, it is dead that ribavirin, the positive group of acyclovir and each concentration group mice of invention medicine part occurred in 5～6 days, positive group mice takes a turn for the better gradually in 7～8 days (after being drug withdrawal 1 day) initial states, activity, food ration increase, and body weight begins to rise.Though and each concentration group mice partial status of invention medicine takes a turn for the better to some extent.
3.2.2.2 the invention medicine is to the influence of mouse lung index
Invention Drug therapy pneumonia the results are shown in Table 5,6.
Resisiting influenza virus effect body weight and lung index compare (n=4) in the table 5 medicine body
As can be seen from Table 5, the invention drug level can effectively improve the lung index greater than 5% o'clock, illustrated that the invention medicine can effectively treat the pneumonia that influenza virus causes.
Anti-adenovirus effect body weight and lung index compare (n=4) in the table 6 medicine body
As can be seen from Table 6, the invention medicine can effectively improve the lung index, illustrates that the invention medicine can effectively treat the pneumonia that adenovirus causes.
To sum up, oil of Herba Pogostemonis has antiviral effect, resistance to influenza virus, CVB-3 and adenovirus is clear and definite, can treat respiratory tract infectious disease, pneumonia, otitis media, viral myocarditis, conjunctivitis, encephalitis, keratitis, hepatitis and enteritis, wherein, to the determined curative effect of viral myocarditis, pneumonia, pharmacological action is strong, and market application foreground is good.
Claims (11)
1. oil of Herba Pogostemonis is in the purposes of preparation in the antiviral drugs.
2. purposes according to claim 1, it is characterized in that: described antiviral drugs is the medicine of resisiting influenza virus, CVB-3 and/or adenovirus.
3. purposes according to claim 2 is characterized in that: described antiviral drugs is that treatment respiratory tract infectious disease, pneumonia, otitis media, viral myocarditis, conjunctivitis, encephalitis, keratitis, hepatitis are or/and the medicine of enteritis.
According to claim 3 for, it is characterized in that: the medicine of described treatment respiratory tract infectious disease is that treatment rhinitis, pharyngitis, tonsillitis, laryngitis, tracheitis are or/and bronchitic medicine.
5. according to any described purposes of claim 1～4, it is characterized in that: described oil of Herba Pogostemonis derives from the volatile oil of Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze. extraction.
6. purposes according to claim 5 is characterized in that: the described oil of Herba Pogostemonis that comes from Herba Pogostemonis Pogostemon cablin (Blanco) Benth., patchouli alcohol content is not less than 40%(w/w), Pogostone content is not less than 20%(w/w).
7. according to any described purposes of claim 1～6, it is characterized in that: described oil of Herba Pogostemonis is to be prepared as follows: the herb of getting Herba Pogostemonis Pogostemon cablin (Blanco) Benth. or Herba Pogostemonis Agastache rugosa (Fisch.Et Mey.) O.Ktze., be ground into coarse powder, add water, soak, the employing steam distillation extracts, namely.
8. antiviral medicinal composition is characterized in that: it is to be active component with the oil of Herba Pogostemonis, adds the medicament that acceptable accessories or complementary composition are prepared from.
9. pharmaceutical composition according to claim 8, it is characterized in that: described medicament is external preparation, oral formulations or ejection preparation.
10. pharmaceutical composition according to claim 9, it is characterized in that: described oral formulations is tablet, capsule, granule.
11. pharmaceutical composition according to claim 8 is characterized in that: the content of oil of Herba Pogostemonis is 0.1%～100%(w/w) in the described preparation.
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| CN116440188A (en) * | 2023-05-17 | 2023-07-18 | 天津中医药大学 | Medical application of wrinkled giant hyssop leaf extract |
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