CN1663604A - Spray for treating chronic pharyngitis - Google Patents
Spray for treating chronic pharyngitis Download PDFInfo
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- CN1663604A CN1663604A CN 200410104428 CN200410104428A CN1663604A CN 1663604 A CN1663604 A CN 1663604A CN 200410104428 CN200410104428 CN 200410104428 CN 200410104428 A CN200410104428 A CN 200410104428A CN 1663604 A CN1663604 A CN 1663604A
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Abstract
The present invention belongs to a spray for treating chronic pharyngitis, especially a spray synchronized by biological preparation and Chinese traditional medicine for treating chronic pharyngitis. It is characterized in that the spray mainly includes Alpha-mannan peptides 10-40%, and antiphlogistic refrigeration ease pain Chinese traditional medicine 60-90%. The component includes Alpha-mannan peptides 10-40%, antiphlogistic refrigeration ease pain Chinese traditional medicine 10-50%, and relieving cough Chinese traditional medicine 10-60%. The component includes Alpha-mannan peptides 10-40%, antiphlogistic refrigeration ease pain Chinese traditional medicine 10-50%, relieving cough Chinese traditional medicine 10-60%, and 10-60% Chinese traditional medicine for reducing capillary permeability. The spray for treating chronic pharyngitis is a Chinese Western medicine composite preparation, is convenient in use and high in local medicine concentration, and can be prescribed reasonably according to the pathological mechanism of the chronic pharyngitis, act against various pathological changes, exert synthetic curative effect and achieve thorough healing.
Description
Affiliated technical field
The invention belongs to a kind of spray for the treatment of chronic pharyngitis, particularly about a kind of spray with biological preparation and the synthetic treatment chronic pharyngitis of Chinese medicine.
Background technology
Chronic pharyngitis is common clinical, frequently-occurring disease.Modern medicine thinks that this disease is that antibacterial, virus and some deleterious physics, chemical factor stimulate the pharyngeal mucosa that causes, sticking touching down and the reaction of adenoid diffuse inflammation.Its histological type can be divided into chronic simple pharyngitis, chronic hypertrophic pharyngitis.Local pathological change is mainly the hyperemia of pharyngeal mucous membrane layer, connective tissue and lymphadenosis under the mucosa, mucous gland hypertrophy, hypersecretion.Chinese medicine thinks that primary disease is many by wind-heat evil toxin invasion and attack bottleneck throat, or the stomach mansion have hot and suffocating, upper punch throat, or hyperactivity of deficient fire causes.The pathogeny of chronic pharyngitis summarize have following some: the 1. pharyngeal infection that pathogenic microorganisms such as antibacterial and virus are arranged; 2. Abwehrkraft des Koepers descends, the comprehensive adjustment ability drop; 3. pharyngeal connective tissue proliferation, sticking touching with peripheral nerve is in high sensitiveness; 4. pharyngeal vascular permeability increases, vascular endothelial cell swelling, luminal stenosis, thrombosis; 5. local immunity reaction imbalance, mastocyte is reactive to be strengthened, and the local immunity complex generates in a large number.
Summary of the invention
The purpose of this invention is to provide a kind of spray for the treatment of chronic pharyngitis, it treats chronic pharyngitis with biological preparation and the synthetic spray of Chinese medicine.This spray is a kind of Chinese medicine and western medicine compound formulation, and it is easy to use, the local drug concentration height, and at the pathomechanism of chronic pharyngitis, scientific composition, the pathological change of antagonism each side, the performance comprehensive therapeutic effect reaches thorough healing.
Technical scheme of the present invention is: design a kind of spray for the treatment of chronic pharyngitis, it is characterized in that: it mainly includes the Chinese medicine 60-90% of α-mannatide 10-40%, antiinflammatory antipyretic-antalgic class.
Described composition includes the Chinese medicine 10-50% of α-mannatide 10-40%, antiinflammatory antipyretic-antalgic class, the Chinese medicine 10-60% of antitussive class.
Described composition includes Chinese medicine 10-40%, the Chinese medicine 10-50% of antitussive class, the Chinese medicine 10-60% of reduction capillary permeability of α-mannatide 10-30%, antiinflammatory antipyretic-antalgic class.
Described composition include Chinese medicine 10-35%, the antitussive class of α-mannatide 10-25%, antiinflammatory antipyretic-antalgic class Chinese medicine 10-40%, reduce capillary permeability Chinese medicine 10-50%, reduce blood viscosity, improve the Chinese medicine 10-40% of hemorheological property.
The Chinese medicine of described antiinflammatory antipyretic-antalgic class: Flos Lonicerae, Radix Isatidis, Fructus Forsythiae, Folium Isatidis, Indigo Naturalis, Rhizoma Coptidis, Radix Scutellariae, Cortex Phellodendri, Caulis Fibraureae, Herba Andrographis, Herba Violae, Herba Taraxaci, Herba Patriniae, Rhizoma Paridis, Radix Gentianae, Radix Sophorae Tonkinensis, the Rhizoma Anemarrhenae, Cortex Magnoliae Officinalis, Cortex Moutan, the Radix Paeoniae Alba, Spica Prunellae, Fructus Trichosanthis, Calculus Bovis, Bulbus Allii, Fructus Chebulae; The Chinese medicine of antitussive class: Radix Rehmanniae, Radix Glycyrrhizae, Bulbus Fritillariae Thunbergii, Fritillary flower, Folium Eriobotryae, Folium Rhododendri Daurici, Folium et Cacumen Rhododendri Mariae, Herba Cymbopogonis, the Rhizoma Pinelliae, Radix Peucedani, Cortex Mori, Fructus Aristolochiae, the Rhizoma Anemarrhenae, Semen Plantaginis, Bulbus Lilii, Radix Ophiopogonis, Rhizoma Polygoni Cuspidati; Reduce the Chinese medicine of capillary permeability: the Flos Sophorae Immaturus, Flos Sophorae, Fructus Forsythiae, Rhizoma Imperatae, the Radix Astragali, Caulis Sargentodoxae, Radix Scutellariae, Cornu Bubali, CORTEX ACANTHOPANACIS, Pericarpium Citri Reticulatae Viride, Pericarpium Citri Reticulatae; Reduce blood viscosity, improve the Chinese medicine of hemorheological property: ligustrazine, Radix Salviae Miltiorrhizae, the Radix Astragali, Radix Rehmanniae, Hirudo, Ganoderma, Radix Puerariae, Flos Carthami, Radix Paeoniae Rubra.
The composition of the every 1000g of described medicament is; α-mannatide 400g, Flos Lonicerae 40g, Radix Isatidis 50g, Fructus Forsythiae 20g, Folium Isatidis 40g, Indigo Naturalis 20g, Rhizoma Coptidis 20g, Radix Scutellariae 20g, Cortex Phellodendri 20g, Caulis Fibraureae 20g, Herba Andrographis 30g, Herba Violae 30g, Herba Taraxaci 40g, Herba Patriniae 10g, Rhizoma Paridis 10g, Radix Gentianae 10g, Radix Sophorae Tonkinensis 30g, Rhizoma Anemarrhenae 20g, Cortex Magnoliae Officinalis 10g, Cortex Moutan 40g, Radix Paeoniae Alba 20g, Spica Prunellae 10g, Fructus Trichosanthis 10g, Calculus Bovis 10g, Bulbus Allii 50g, Fructus Chebulae 20g.
The composition of the every 1000g of described medicament is; α-mannatide 365g, Flos Lonicerae 20g; Radix Isatidis 10g; Fructus Forsythiae 20g; Folium Isatidis 15g; Indigo Naturalis 20g; Rhizoma Coptidis 10g; Radix Scutellariae 15g; Cortex Phellodendri 20g; Caulis Fibraureae 15g; Herba Andrographis 20g; Herba Violae 15g; Herba Taraxaci 25g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 20g; Rhizoma Anemarrhenae 15g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 25g; Radix Paeoniae Alba 20g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 30g; Fructus Chebulae 20g; Radix Rehmanniae 10g; Radix Glycyrrhizae 20g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 25g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g.
The composition of the every 1000g of described medicament is; α-mannatide 250g, Flos Lonicerae 20g; Radix Isatidis 20g; Folium Isatidis 15g; Indigo Naturalis 10g; Rhizoma Coptidis 10g; Cortex Phellodendri 10g; Caulis Fibraureae 15g; Herba Andrographis 10g; Herba Violae 15g; Herba Taraxaci 15g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 15g; Rhizoma Anemarrhenae 15g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 25g; Radix Paeoniae Alba 10g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 20g; Fructus Chebulae 15g; Radix Rehmanniae 10g; Radix Glycyrrhizae 20g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 25g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g; Flos Sophorae Immaturus 20g; Flos Sophorae 20g; Fructus Forsythiae 30g; Rhizoma Imperatae 20g; Radix Astragali 15g; Caulis Sargentodoxae 20g; Radix Scutellariae 20g; Cornu Bubali 20g; CORTEX ACANTHOPANACIS 20g; Pericarpium Citri Reticulatae Viride 10g; Pericarpium Citri Reticulatae 15g.
The composition of the every 1000g of described medicament is; α-mannatide 200g, Flos Lonicerae 20g; Radix Isatidis 10g; Folium Isatidis 15g; Indigo Naturalis 10g; Rhizoma Coptidis 10g; Cortex Phellodendri 10g; Caulis Fibraureae 15g; Herba Andrographis 10g; Herba Violae 10g; Herba Taraxaci 15g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 15g; Rhizoma Anemarrhenae 10g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 15g; Radix Paeoniae Alba 10g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 15g; Fructus Chebulae 15g; Radix Rehmanniae 10g; Radix Glycyrrhizae 15g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 15g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g; Flos Sophorae Immaturus 20g; Flos Sophorae 20g; Fructus Forsythiae 30g; Rhizoma Imperatae 20g; Caulis Sargentodoxae 20g; Radix Scutellariae 20g; Cornu Bubali 20g; CORTEX ACANTHOPANACIS 20g; Pericarpium Citri Reticulatae Viride 10g; Pericarpium Citri Reticulatae 15g; ligustrazine 10g; Radix Salviae Miltiorrhizae 10g; Radix Astragali 30g; Radix Rehmanniae 10g; Hirudo 10g; Ganoderma 10g; Radix Puerariae 15g; Flos Carthami 10g; Radix Paeoniae Rubra 10g.
Characteristics of the present invention are: α-mannatide is a kind of biological response modifier, can make too high or too low immunoreation recover normal, and the enhancing body resistance also has antiphlogistic effect; Flos Lonicerae, Radix Isatidis have antiinflammatory, analgesia, antipyretic effect; Radix Rehmanniae, Radix Glycyrrhizae have inhibitory action to the connective tissue proliferation in inflammation later stage, and have sticking the touching of the pharyngeal and bronchus of protection and avoid stimulating, and act on throat's peripheral nerve weakening cough, thereby reach cough suppressing effect; Radix Scrophulariae, Herba Menthae increase capillary permeability the obvious suppression effect, thereby suppresses inflammatory exudation; Radix Trichosanthis has the expansion local vascular, improves the blood stasis state, and can suppress or alleviate the infiltration of inflammatory cell; Fructus Ligustri Lucidi, Radix Angelicae Sinensis, Radix Platycodonis remove has the whole blood of reduction and plasma viscosity, improve hemorheological property, promote the absorption of local inflammation, also have immunoregulatory effect, can significantly suppress mastocyte and under antigen is attacked, take off the granule reaction, strengthen the immune complex ability in the blood circulation of removing.
The local excitation test of treatment chronic pharyngitis spray of the present invention: select 6 of 2.5-3.0kg healthy rabbits, this spray of topical application, wherein 3 disposable heavy dose of contacts, partial reacting phenomenon behind observation and record 24h, the 72h.The result shows: do not see local excitation reactions such as rubescent, dermexanthesis, vesicle behind medication 24h, the 72h, illustrate that treatment chronic pharyngitis spray of the present invention does not have the acute irritation reaction; Other 3 every days three times, each low dose of contact continues to observe after 10 days, does not see local excitation reactions such as rubescent, dermexanthesis, vesicle yet, illustrates that treatment chronic pharyngitis spray of the present invention does not have chronic cumulative bad irritant reaction.This spray is to be perfectly safe, and do not have any untoward reaction.
The specific embodiment
Because the present invention is carried recoverable space craft (spacecraft or retrievable satellite) with the Alpha-hemolytic streptococcus strain, utilize the comprehensive function of factors such as little in the space (zero) gravity, cosmic ray, alternating magnetic field, fine vacuum, hyperpyrexia deep cooling, make the dna double chain structure fracture of bacterial strain, genome is reset, thereby produces new bacterial strain.After returning ground, bacterial strain through space treatment is cultivated and screened, by studying contents such as its morphological characteristic, cultural characteristic, biochemical reactions, metabolite, hereditary character and protein expression, hereditary stability, pilot scale production target, optimize the stable strain of plus variant, inherited character wherein, the medicine of the ulcerative colitis that after cultivation and fermentation, obtains medical treatment.This strain in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, is numbered CGMCCNo.1082.
Particularly Alpha-hemolytic streptococcus D33 bacterial strain behind ground screening, separation and the purification, selects higher, the secreted mannan peptide content of fermentation unit higher high yield, quality strains T33 strain through space treatment mutation.By Institute of Microorganism, Academia Sinica whole-cell protein SDS-polyacrylamide gel (SDS-PAGE) analysis of T33 bacterial strain and D33 bacterial strain and the genomic DNA restricted enzyme cutting analysis (RFLP/PFGE) of bacterial strain are compared, can prove that its gene of T33 bacterial strain that space mutagenesis and ground filter out has variation.This mutant strain is stable in heredity, stronger production adaptability is arranged, the mannan peptide content that is produced by its fermentation improves, content of peptides improves 3 to 5 times than national standard (80%), institute for drug control, Xi'an sample presentation is measured and is reached 271.6%, and fermentation content on average exceeds more than three times than matched group product content.This strain is through the spaceship-carried space flight of Shenzhou series of spacecraft, and further screening and culturing breeding forms stable hereditary property, the tangible high-yield highly-effective strain of variation features to strain by BeiJing ZhongKe Institute of Micro-biology of institute and biology department of Northwest University.The mannan peptide content of α in the every fermentation unit of this strain-space flight strain production has improved more than several times.(the material 1 of seeing Appendix: science and technology bureau Shaanxi Province, Shaanxi Province scientific and technological achievement assay certificate material
Adnexa material 2: the mannatide that the space flight strain is produced is produced bacterial strain and is carried notarization through " No. three, divine boat " airship.
Adnexa material 3: the mannatide that the space flight strain is produced is produced bacterial strain and is returned ground screening report through the lift-launch of " No. three, divine boat " airship
Adnexa material 4: Microbe Inst., Chinese Academy of Sciences produces the form Physiology and biochemistry probation report book of bacterial strain and ground contrast bacterium about the mannatide that " No. three, divine boat " airship carries the production of space flight strain
Adnexa material 5: Microbe Inst., Chinese Academy of Sciences produces SDS-PAGE and the RFLP/PFGE Analysis and Identification statement of bacterial strain and ground contrast bacterium about the mannatide that " No. three, divine boat " airship carries the production of space flight strain
Adnexa material 6: the mannatide oral liquid examining report that institute for drug control, Shaanxi Province " No. three, divine boat " space flight strain is produced
Adnexa material 7: scientific and technological novelty assessment report copy
Adnexa material 8: notice is accepted in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation culture presevation)
Strain improvement of the present invention: on the basis of Alpha-hemolytic streptococcus growth rhythm, selected than the proper growth strain in period, adopt the plate growth bacterium colony, immerse methods such as the bacteria suspension in other material, germ-carrying sand, carry " No. three, divine boat " spacecraft on March 25th, 2002.Carried out 6 days 0 18 hours flying at space track (200 kilometers of perigee altitudes, 350 kilometers of altitude of the apogees) at rail.The specific condition of space mainly is presented as special environments such as microgravity, fine vacuum, high radiation, alternating magnetic field.The strain of outer-space flight be placed in one with the diverse environment of tellurian ecological environment in, the electronics, proton and the mental retardation heavy particle that mainly comprise coming self-magnetic field to capture; Cosmic ray such as proton, particle and heavier high energy heavy particle from the milky way galaxy; From the proton of sun magnetic storm and heavy particle etc.These particles act on the dna double chain structure in the microbial cell nuclear, can cause double-strand break.Microgravity, high vacuum environment can make the base sequence of DNA recombinate, promptly genomic reorganization and variation.The new bacterial strain that the variation back produces, variation in various degree all can take place in its morphological characteristic, cultural characteristic, biochemical reactions, metabolite, hereditary character and protein expression, pilot scale production target etc.After ground is returned in spacecraft, through further screening, only optimize wherein 0.2% plus variant and the stable bacterial strain of inherited character, make the production strain through cultivation.This strain that after spaceship-carried mutation, selects December in 2003 29 days by China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, be numbered CGMCCNo.1082.
Experiment and middle trial production result show, stable on inherited character through the novel space strain behind the space mutagenesis, stronger production adaptability is arranged, the mannan peptide content that is produced by its fermentation improves, content of peptides improves 3 to 5 times than national standard (80%), institute for drug control, Xi'an sample presentation is measured and is reached 271.6%, and fermentation content on average exceeds more than three times than matched group product content, and fermentation period shortens greatly.
Preparation method:
The space flight strain is produced the preparation process of mannatide:
The mannatide of space flight strain of the present invention production is by following method preparation: the space strain preparation--purify, and final production goes out mannatide by one grade fermemtation--second order fermentation-----fermentation liquid.
space strain preparation:
With through the Alpha-hemolytic streptococcus of space mutagenesis breeding meticulous selection-breeding as producing strain.
A. inclined-plane seed culture medium: Carnis Bovis seu Bubali cream 0.5%, yeast extract 0.6%, peptone 0.5%, glucose 0.4%, sodium chloride 0.5%, agar 3%, sheep blood 8%; PH7.4.
Inclined-plane seed culture medium sterilising temp 121,30 minutes time, steam pressure 0.12Mpa.
Unpacking strain cryovial with aseptic broth bouillon dilution, inserts the blood inclined-plane under aseptic condition, inoculate rearmounted 38 constant temperature culture 30 hours.
B. meat soup seed culture medium: Carnis Bovis seu Bubali cream 0.5%, yeast extract 0.6%, peptone 0.5%, glucose 0.4%, sodium chloride 0.5%; PH7.4.
Meat soup seed culture medium sterilising temp 121,30 minutes time, steam pressure 0.12Mpa.
Cultured slant strains is pressed 9% inoculum concentration under aseptic condition, insert in the meat soup seed culture medium 38 constant temperature culture 30 hours.
The sweat:
Fermentation medium: Carnis Bovis seu Bubali cream 0.4%, yeast extract 0.5%, peptone 0.5%, glucose 0.4%, sodium chloride 0.5%.
Fermentation medium sterilising temp 121,30 minutes time, steam pressure 0.12Mpa.
A. one grade fermemtation jar: with good meat soup strain under aseptic condition by 10% inoculum concentration, insert first class seed pot, 29 constant temperature culture 30 hours, tank pressure was no more than 0.02Mpa, ventilation is advisable can stir culture fluid, fully stirs continuously.
B. second order fermentation jar: with the one grade fermemtation culture fluid under aseptic condition by 20% inoculum concentration, insert fermentation tank, 29 constant temperature culture 70 hours, tank pressure is 0.02Mpa not, jar is put in deactivation.Deactivation is adopted to heat and is made the fermentation liquid temperature reach 100, is incubated 60 minutes, and cooling is left standstill.
The leaching process:
A, fermentation liquid concentrate, and make concentrated solution volume and fermentating liquid volume ratio be controlled at 1: 15-1: 20 or be determined by circumstances.
The concentrated solution of b, fermentation liquid adds 80-99% ethanol, and the volume ratio is controlled at 1: 1.5-1: 5.5 or be determined by circumstances.Fully stir leave standstill after, centrifugal removal supernatant, the precipitation dissolved in distilled water, accent pH5.0 obtains lysate and lysate is left standstill.
C, the more centrifugal removal impurity of lysate is obtained supernatant, accurately measure the clear liquid volume, calculate required amount of alcohol by the supernatant stereometer, adjust pH slowly joins ethanol in the supernatant, and fully stirs, and leaves standstill the centrifugal removal supernatant in back and obtains precipitate.
D, precipitate reuse dissolved in distilled water are transferred pH, and centrifugal, supernatant adds ethanol precipitation again.Such technology repeatable operation to intermediate detection qualified till, the precipitate that obtains is the mannatide crude product that the space flight strain is produced.Vacuum drying 3-8 hour, promptly obtain the mannan peptide product that the space flight strain is produced.
The check of the product that said method obtains:
[character] this product is white or little yellow amorphous powder; Odorless, tasteless.
This product is easily molten in water, and is insoluble in ethanol, chloroform and acetone.
Specific optical rotation: get this product, accurate claim surely, be dissolved in water and be diluted to the solution that contains 10mg among every 1ml approximately, measure (two appendix vIE of Chinese Pharmacopoeia version in 2000) in accordance with the law, specific optical rotation be extremely+80 of+70.
[discriminating] 1, get this product 10mg, add water 0.5ml and make dissolving, add a-naphthols alcoholic solution (5-100) 1ml, shake up, slowly add sulphuric acid 0.5ml along tube wall, after several minutes, the interface is aubergine.
2, get this product, add water and make the solution that contains 1mg among every 1ml, get about 10ul point on filter paper, dry, fix, put high salpeter solution and (get periodic acid 1.2g with dehydrated alcohol, after adding water 30ml dissolving. add 0.2mol/L sodium acetate solution 1.5ml and ethanol 100ml, mixing is promptly.Put the dark place and preserve, can use the several months) the middle immersion 5 minutes, take out, wash with 70% alcoholic solution, (get potassium iodide 5g, sodium thiosulfate 5g adds water 100ml and makes dissolving to put reducing solution, add ethanol 150ml, 2mol/L hydrochloric acid solution 2.5ml again, with adding, face the time spent and join with stirring) the middle immersion 5 minutes, take out, wash with 70% alcoholic solution, put in the pinkish red industry sulphuric acid test solution and soaked about 30 minutes, take out, (get sodium pyrosulfite 0.4g with sodium metabisulfite solution, add hydrochloric acid 1ml, being dissolved in water makes into 100ml, promptly), and flushing, dry, the place should be aubergine at the filter paper point sample.
3, get test sample and reference substance is an amount of, add respectively the chlorination sodium injection make contain 1mg among every 1ml solution as need testing solution and reference substance solution, press the test of mannatide immunogenicity determining method, test sample and contrast QC should all not have haemolysis to be taken place.
[inspection] 1, acidity: get this product, add water and make the solution that contains 10mg among every 1ml, measure (two appendix of Chinese Pharmacopoeia version in 2000 H) in accordance with the law, pH value should be 3.0-5.0.
2, trap: get this product, add water and make the solution that contains 0.4mg among every 1ml, according to spectrophotography (two appendix of Chinese Pharmacopoeia version in 2000 A), wavelength place at 260nm, its trap must not be greater than 0.25, and at the wavelength place of 280nm, its trap must not be greater than 0.20.
3, total nitrogen: get this product, measure according to N2 method (two appendix of Chinese Pharmacopoeia version in 2000 D, second method). press dry product and calculate, contain total nitrogen and should be 0.8-2.0%.
4, immunogenicity: get test sample and reference substance is an amount of, add the chlorination sodium injection respectively and make the solution that contains 10mg among every 1ml, make 1: 2,1: 4,1: 8,1: 16,1: 32,1: 64,1: 128,1: 256 diluent as need testing solution and reference substance solution with phosphate buffer respectively again, check in accordance with the law (attached mannatide immunogenicity determining method) that the least concentration of the insoluble blood vessel of test sample should be higher than more than a times of reference substance respective concentration.
5, loss on drying is got this product, is dried to constant weight at 105, subtracts weight loss and must not cross 5.0% (two appendix of Chinese Pharmacopoeia version in 2000 L).
6, heavy metal is got this product, at 1.0g, checks that in accordance with the law (Chinese Pharmacopoeia version in 2000 H) contains heavy metal and must not cross 20/1000000ths.
7, the undue toxicity gets this product, adds the chlorination sodium injection and makes the solution that contains 0.5mg among every 1ml, checks (Chinese Pharmacopoeia version appendix in 2000 C) in accordance with the law. press the intravenous injection administration, and should (injection) up to specification.
[assay]
1. the preparation of reference substance solution
Precision takes by weighing the D-mannose reference substance 0.1g that is dried to constant weight through 105 and puts in the 100ml measuring bottle, is dissolved in water and is diluted to scale. shake up; Precision is measured 5ml, puts in the measuring bottle of 100ml, adds water to scale, shakes up. contain mannose 50ug among every 1ml.
2. need testing solution is equipped with
It is an amount of to get this product, and accurate the title decides, and is dissolved in water and makes the solution that contains 40ug among every 1ml.
3. the preparation of standard curve
Precision takes by weighing reference substance solution 0,0.2,0.4,0.6,0.8,1.0ml, put respectively in the tool plug test tube, respectively add water to 1.0ml, add 3% phenol solution 1.0ml again, shake up, pour sulphuric acid 4.5ml, shake up, being positioned over room temperature, is blank with 0 pipe. measure trap according to spectrophotography (two appendix of Chinese Pharmacopoeia version in 2000 A) at the wavelength place of 490nm.To corresponding trap, calculate regression equation with mannose ug number.
4. algoscopy
Precision is measured need testing solution 1.0ml, and from " adding 3% phenol solution 1.0ml again ", operation is in accordance with the law measured trap, by the content of regression equation calculation mannose under the sighting target directrix curve preparation.
Mannatide immunogenicity determining method (complement combined techniques);
1, test solution
A. phthalate buffer (pH7.2)
Get sodium chloride 8.5g, sodium hydrogen phosphate 0.565g and potassium dihydrogen phosphate 0.135g, add water to 1000ml and make its dissolving, add 10% Adlerika 1ml, shake up.
B.1% sheep erythrocyte suspension
The preparation of sheeps blood erythrocyte: (get glucose 20.5g, sodium citrate 8.0g, sodium chloride 4.2g in filling equivalent A Shi liquid by sheep jugular vein sterile blood sampling, citric acid 5.5g, add water to 1000ml and make dissolving, 100 sterilization 30 minutes) sterile chamber in, 4 preserve.
The preparation of 1% sheeps blood erythrocyte suspension: it is an amount of to get above-mentioned sheeps blood erythrocyte, and with sodium chloride injection washing three times, each centrifugal 5 minutes (2000 rev/mins), sheeps blood erythrocyte is amassed in pressure, makes 1% sheeps blood erythrocyte suspension with sodium chloride injection.Get suspension, make need testing solution for 20 times with the sodium chloride injection dilution, other gets 20 times of equivalent suspension thin ups as blank solution, according to spectrophotography (two appendix of Chinese Pharmacopoeia version in 2000 A), wavelength place at 541nm measures, its trap should be 0.65-0.70, should regulate the concentration of 1% sheep erythrocyte suspension as overrun.
C. hemolysin and sensitization sheeps blood erythrocyte
The preparation of hemolysin: get above-mentioned hematocrit sheeps blood erythrocyte, make 25% sheeps blood erythrocyte suspension with sodium chloride injection. get 1 of rabbit, the above-mentioned sheeps blood erythrocyte suspension of intravenous injection, once a day, totally seven times, first three time 3ml, three 5ml in back, last is injected blood sampling in back seven days.Separation of serum, 56 .30 minute deactivations, packing, 0 is following to be preserved.
The mensuration of amboceptor unit: it is an amount of to get hemolysin, add phosphate buffer and make 1: 1000,1: 2000,1: 3000,1: 4000,1: 5000,1: 6000,1: 7000,1: 8000,1: 9000,1: 10000 diluent respectively, respectively getting 0.1ml puts in the test tube, add 1% Sanguis caprae seu ovis cell suspension 0.1ml, shake up, add dilution factor and be 1: 30 guinea pig serum (complement) 0.2ml and phosphate buffer 0.2ml, shake up, 37 insulation 30 minutes, the high dilution of complete hemolysis pipe is 1 unit hemolysin.
The preparation of sensitization sheeps blood erythrocyte: before facing usefulness, the sheeps blood erythrocyte suspension with 2% mixes with 2 unit haemolysis rope equal-volumes, and 37 are incubated 15 minutes promptly.
Complement: get the Cavia porcellus more than three, heart blood sampling centrifugalize serum, 0 degree is following to be preserved.
The mensuration of complement unit: it is an amount of to get complement, add phosphate buffer, make 1: 40,1: 60,1: 80,1: 100,1: 120,1: 140,1: 160,1: 180 diluent respectively, respectively get 0.2ml and put in the test tube, add the 0.2ml phosphate buffer, shake up, 37 insulation is after 30 minutes, add sensitization sheeps blood erythrocyte 0.2ml respectively, shake up, 37 are incubated 30 minutes again. and the high dilution of complete hemolysis pipe is the complement of 1 unit.
Antibody: it is an amount of to get the mannatide reference substance, add the chlorination sodium injection and make the reference substance solution immunizing rabbit that contains 10mg among every 1ml, the next day adopt ear vein injection reference substance solution once.Inject for the first time 0.2ml, for the second time respectively inject 0.5ml to the 5th time, respectively inject 1.0ml the 6th time to the tenth time, the tenth once respectively injects 2.0ml to the 15 time, and last is injected blood sampling in back three days, centrifugalize serum, 56 deactivation in following 30 minutes, packing, 0 is following to be preserved (before facing usefulness, needing 56 deactivation once more in 30 minutes).
The mensuration of antibody unit: it is an amount of to get antibody, add phosphate buffer and make 1: 2,1: 4,1: 8,1: 16,1: 32.1: 64,1: 128 diluent respectively, respectively getting 0.1ml puts in the test tube, add mannatide reference substance solution 0.1ml and the 2 complement 0.2ml of unit, shake up, place more than 4 hours in 4-8, put 37 insulation 30 minutes, the high dilution of insoluble blood vessel is 1 unit antibody.Establish antigen simultaneously and (do not add antibody, replace with the 0.1ml phosphate buffer), antibody (do not add antigen, replace with the 0.1ml phosphate buffer), complement (do not add antigen, antibody, replace with the 0.2ml phosphate buffer) control tube, more than three kinds of control tube haemolysis entirely; The sensitization sheeps blood erythrocyte that other establishes (do not add antigen, antibody and complement, replace with the 0.4ml phosphate buffer) control tube is haemolysis not.
2, immunogenicity determining method
It is an amount of to get rapid glycopeptide reference substance of manna and test sample, make the reference substance solution and the need testing solution of variable concentrations according to the regulation under the medicine item, respectively getting 0.1ml puts in the test tube, add the 2 antibody 0.1ml of unit and the 2 complement 0.2ml. of unit shake up, place more than 4 hours in 4-8, put 37 insulation 30 minutes, add sensitization sheeps blood erythrocyte 0.2ml, shake up, 37 are incubated 30 minutes again.Observe the haemolysis situation of each pipe, the least concentration of insoluble blood vessel is represented the immunogenicity of mannatide.Establish antigen simultaneously and (do not add antibody, replace with the 0.1ml phosphate buffer), antibody (do not add antigen, replace with the 0.1ml phosphate buffer), complement (do not add antigen, antibody, replace) control tube with the 0.2ml phosphate buffer, more than three kinds of control tube haemolysis entirely: the sensitization sheeps blood erythrocyte that other establishes (do not add antigen, antibody and complement, replace with the 0.4ml phosphate buffer) control tube is haemolysis not.
Embodiment 1 prescription is formed: get above-mentioned α-mannatide 500g, and Flos Lonicerae 150g, Radix Isatidis 150g, Radix Rehmanniae 120g, Radix Glycyrrhizae 120g, Radix Scrophulariae 120g, Herba Menthae 120g, Radix Trichosanthis 100g, Fructus Ligustri Lucidi 120g, Radix Angelicae Sinensis 150g, Radix Platycodonis 100g makes every bottle of 100ml.
During preparation with above Chinese medicine dense fry in shallow oil three times → merge medicine juice → filtrations → concentrated → with α-mannatide mixing → filtration → quality inspection → packing → finished product qualified back → warehouse-in to be checked.The preparation technology that concrete production process is pressed 2000 editions sprays of Pharmacopoeia of People's Republic of China carries out.(Chinese Pharmacopoeia Commission compiles; Chemical Industry Press publishes)
Other embodiment is identical with the production process of embodiment 1, just the drug component difference.
Contain α-mannatide 400g among the medicine 1000g of embodiment 2, Flos Lonicerae 40g, Radix Isatidis 50g, Fructus Forsythiae 20g, Folium Isatidis 40g, Indigo Naturalis 20g, Rhizoma Coptidis 20g, Radix Scutellariae 20g, Cortex Phellodendri 20g, Caulis Fibraureae 20g, Herba Andrographis 30g, Herba Violae 30g, Herba Taraxaci 40g, Herba Patriniae 10g, Rhizoma Paridis 10g, Radix Gentianae 10g, Radix Sophorae Tonkinensis 30g, Rhizoma Anemarrhenae 20g, Cortex Magnoliae Officinalis 10g, Cortex Moutan 40g, Radix Paeoniae Alba 20g, Spica Prunellae 10g, Fructus Trichosanthis 10g, Calculus Bovis 10g, Bulbus Allii 50g, Fructus Chebulae 20g.
Contain α-mannatide 365g among the medicine 1000g of embodiment 3, Flos Lonicerae 20g; Radix Isatidis 10g; Fructus Forsythiae 20g; Folium Isatidis 15g; Indigo Naturalis 20g; Rhizoma Coptidis 10g; Radix Scutellariae 15g; Cortex Phellodendri 20g; Caulis Fibraureae 15g; Herba Andrographis 20g; Herba Violae 15g; Herba Taraxaci 25g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 20g; Rhizoma Anemarrhenae 15g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 25g; Radix Paeoniae Alba 20g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 30g; Fructus Chebulae 20g; Radix Rehmanniae 10g; Radix Glycyrrhizae 20g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 25g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g.
Contain α-mannatide 250g among the medicine 1000g of embodiment 4, Flos Lonicerae 20g; Radix Isatidis 20g; Folium Isatidis 15g; Indigo Naturalis 10g; Rhizoma Coptidis 10g; Cortex Phellodendri 10g; Caulis Fibraureae 15g; Herba Andrographis 10g; Herba Violae 15g; Herba Taraxaci 15g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 15g; Rhizoma Anemarrhenae 15g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 25g; Radix Paeoniae Alba 10g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 20g; Fructus Chebulae 15g; Radix Rehmanniae 10g; Radix Glycyrrhizae 20g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 25g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g; Flos Sophorae Immaturus 20g; Flos Sophorae 20g; Fructus Forsythiae 30g; Rhizoma Imperatae 20g; Radix Astragali 15g; Caulis Sargentodoxae 20g; Radix Scutellariae 20g; Cornu Bubali 20g; CORTEX ACANTHOPANACIS 20g; Pericarpium Citri Reticulatae Viride 10g; Pericarpium Citri Reticulatae 15g.
Contain α-mannatide 200g among the medicine 1000g of embodiment 5, Flos Lonicerae 20g; Radix Isatidis 10g; Folium Isatidis 15g; Indigo Naturalis 10g; Rhizoma Coptidis 10g; Cortex Phellodendri 10g; Caulis Fibraureae 15g; Herba Andrographis 10g; Herba Violae 10g; Herba Taraxaci 15g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 15g; Rhizoma Anemarrhenae 10g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 15g; Radix Paeoniae Alba 10g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 15g; Fructus Chebulae 15g; Radix Rehmanniae 10g; Radix Glycyrrhizae 15g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 15g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g; Flos Sophorae Immaturus 20g; Flos Sophorae 20g; Fructus Forsythiae 30g; Rhizoma Imperatae 20g; Caulis Sargentodoxae 20g; Radix Scutellariae 20g; Cornu Bubali 20g; CORTEX ACANTHOPANACIS 20g; Pericarpium Citri Reticulatae Viride 10g; Pericarpium Citri Reticulatae 15g; ligustrazine 10g; Radix Salviae Miltiorrhizae 10g; Radix Astragali 30g; Radix Rehmanniae 10g; Hirudo 10g; Ganoderma 10g; Radix Puerariae 15g; Flos Carthami 10g; Radix Paeoniae Rubra 10g.
The animal pharmacodynamic study
Experiment one: the antiinflammatory of spray of the present invention and immunoregulation effect
The four kinds of sprays of the present invention of making a summary are mainly used in the treatment chronic pharyngitis.Pharmacological research proves, four kinds of sprays of the present invention have certain antiinflammatory action, can significantly suppress the take off granule of mastocyte under antigen is attacked reacts, the mastocyte that antigen is caused discharged sensitive media inhibitory action, can improve and infect Sendai virus mouse red blood cell garland formation rate, strengthen the ability that mice is removed immune complex in the blood circulation, the periphery blood T lymphocyte lipase rate of dyeing that antagonism virus causes descends.
1 experiment material
1.1 medicine and reagent
JINHOUJIAN PENWUJI: Hongyu Pharmaceutical Co., Ltd., Guizhou Prov. provides.
α-mannatide spray: provide by Xi'an Hengtongguanghua Pharmaceutical Co., Ltd.
Four kinds of sprays of the present invention: provide by Xi'an Hengtongguanghua Pharmaceutical Co., Ltd.
Sendai virus: provided by microorganism parasite teaching and research room of Xi'an Jiaotong University Medical College, the blood clotting titre is 1: 320.
Chicken erythrocyte (CRBC): come high chicken heparanase anticoagulation ,-4 ℃ are kept in the A Shi liquid, use in 2 weeks.
1.2 animal
Kunming mouse, body weight (20 ± 2) g; SD rat and Wistar rat, body weight (180 ± 20) g, the male and female dual-purpose provides by Xi'an Jiaotong University Medical College's Experimental Animal Center.
2 methods and result
2.1 the antiinflammatory action of four kinds of sprays of the present invention
2.1.1 Oleum Tiglii is brought out the influence of mice auricular concha inflammation
Get 70 mices, be divided into 7 groups at random, 10 every group.Be normal saline matched group, spray group of the present invention, α-mannatide spray group, JINHOUJIAN PENWUJI group.More than each group give four kinds of normal saline, sprays of the present invention, α-mannatide spray, the JINHOUJIAN PENWUJI of equivalent (1ml), every day 3 times, logotype 3 days respectively at every turn.After the last administration 30 minutes, with 2% Oleum Tiglii (its composition: Oleum Tiglii 0.2ml, 95% ethanol 2ml and ether 7.8ml) 0.05ml spreads upon the positive and negative two sides of mice left side ear equably, and auris dextra is measured two ear weight differences (the results are shown in Table 1) in contrast after 4 hours.
Table 1 four kinds of sprays of the present invention bring out mice to Oleum Tiglii
The influence of auricular concha inflammation (X ± s, n=10)
Group two ear weight difference (mg) suppression ratio (%)
Normal saline matched group 18.24 ± 4.11
First kind of spray group 9.72 ± 3.35** 46.71 of the present invention
Second kind of spray group 9.50 ± 2.15** 49.58 of the present invention
The third spray group 9.34 ± 2.27** 51.24 of the present invention
The 4th kind of spray group 9.07 ± 2.08** 54.19 of the present invention
α-mannatide spray group 13.84 ± 3.82* 30.50
JINHOUJIAN PENWUJI group 13.69 ± 4.06* 25.29
Annotate: compare with the normal saline matched group:
*P＜0.05,
*P＜0.01
The experimental data of last table shows: four kinds of sprays of the present invention all can significantly suppress the mice auricular concha inflammation that Oleum Tiglii brings out, and along with the reinforcement of filling a prescription, effect is obvious further; α-mannatide spray and JINHOUJIAN PENWUJI also can suppress the mice auricular concha inflammation that Oleum Tiglii brings out to a certain extent, but suppression ratio is starkly lower than four kinds of sprays of the present invention, illustrate that four kinds of sprays of the present invention have unique therapeutic effect to the mice local inflammation, list α-mannatide and the JINHOUJIAN PENWUJI of being better than evident in efficacy.
2.1.2 influence to the mouse peritoneal capillary permeability
Get 70 mices, be divided into 7 groups at random, 10 every group.Be that normal saline matched group, four kinds of sprays of the present invention form four groups, α-mannatide spray group, JINHOUJIAN PENWUJI group.More than each group give normal saline, four kinds of sprays of the present invention, α-mannatide spray, the JINHOUJIAN PENWUJI of equivalent (1ml), every day 3 times, logotype 3 days respectively at every turn.After the last administration 60 minutes, the blue 5ml/kg of each caudal vein injection 0.5%Evans, pneumoretroperitoneum was injected 0.5% acetic acid 10ml/kg in 5 minutes, and the blue concentration (the results are shown in Table 2) of Evans with the mice sacrificed by exsanguination, was measured with UV-754 type ultraviolet-uisible spectrophotometer after 30 minutes in the interval at the 610mm place.
Table 2 four kinds of sprays of the present invention are to the mouse peritoneal blood capillary
The influence of permeability (X ± s, n=10)
Blue concentration (mg/L) suppression ratio (%) of group Evans
Normal saline matched group 9.51 ± 2.04
First kind of spray group 5.14 ± 1.92* 46.00 of the present invention
Second kind of spray group 5.00 ± 1.63* 48.12 of the present invention
The third spray group 4.86 ± 1.37* 51.69 of the present invention
The 4th kind of spray group 4.50 ± 1.12* 57.03 of the present invention
α-mannatide spray group 7.89 ± 2.73 20.48
JINHOUJIAN PENWUJI group 8.02 ± 2.58 18.38
Annotate: compare with the normal saline matched group:
*P＜0.05
Last table data show: four kinds of sprays of the present invention can significantly suppress the permeability of mice blood capillary, and along with the reinforcement of filling a prescription, effect is obvious further; α-mannatide spray and JINHOUJIAN PENWUJI also have inhibitory action to the permeability of mice blood capillary, but suppression ratio is starkly lower than four kinds of sprays of the present invention.Illustrate that four kinds of sprays of the present invention increase the therapeutic effect that uniqueness is arranged to the mice capillary permeability, list α-mannatide and the JINHOUJIAN PENWUJI of being better than evident in efficacy.
2.1.3 cotton balls is brought out the influence of rat granulation tissue hyperplasia
Get 70 of male SD rats, be divided into 7 groups at random, 10 every group.Be that normal saline matched group, four kinds of sprays of the present invention form four groups, α-mannatide spray group, JINHOUJIAN PENWUJI group.Under aseptic condition, in the rat back otch, sterilized cotton ball 20mg is implanted the oxter, both sides, to sew up the incision, the part is coated with (12.5 * 10
5) kanamycin of U/L.From burying the same day of cotton balls, organize normal saline, four kinds of sprays of the present invention, α-mannatide spray, the JINHOUJIAN PENWUJI of equivalent (1ml), every day 3 times, logotype 8 days every day respectively to each.Pulled cervical vertebra on the 8th day and put to death, take out cotton balls 60 ℃ of oven dry, weigh, the weight that deducts cotton balls is the net weight of granulation tissue (the results are shown in Table 3).
The experimental data of table 3 shows: four kinds of sprays of the present invention can significantly suppress to bury the rat granuloma that cotton balls brings out, and along with the reinforcement of filling a prescription, effect is obvious further; α-mannatide spray and JINHOUJIAN PENWUJI also have inhibitory action to the rat granuloma of burying cotton balls and bringing out, but suppression ratio is starkly lower than four kinds of sprays of the present invention.Illustrate that four kinds of sprays of the present invention have unique therapeutic effect to the rat granuloma of burying cotton balls and bringing out, evident in efficacy be better than single with α-mannatide and JINHOUJIAN PENWUJI.
Table 3 four kinds of sprays of the present invention bring out rat to burying cotton balls
Granulomatous influence (X ± s, n=10)
Group granuloma thousandweight (mg) suppression ratio (%)
Normal saline matched group 80.19 ± 11.71
First kind of spray group 51.45 ± 10.86* 40.83 of the present invention
Second kind of spray group 49.13 ± 9.27* 43.04 of the present invention
The third spray group 47.38 ± 9.02* 45.68 of the present invention
The 4th kind of spray group 45.06 ± 8.32* 49.78 of the present invention
α-mannatide spray group 71.89 ± 12.73 10.48
JINHOUJIAN PENWUJI group 78.62 ± 10.58 4.37
Annotate: compare with the normal saline matched group:
*P＜0.05
2.2 spray of the present invention is to Immune Effects
2.2.1 influence to the reaction of rat cranial periosteum mast cell degranulation
Get 70 of Wistar rats, be divided into 7 groups at random, promptly normal saline matched group, four kinds of sprays of the present invention form four groups, α-mannatide spray group, isoproterenol group (50 μ g/kg are made into 1ml solution).Each is organized and gives normal saline, four kinds of sprays of the present invention, α-mannatide spray, isoproterenol every day respectively 3 times, be total to 1ml, continuous 6 days, after the last administration, the blue solution of 1: 20 rat anti trichosanthin antiserum 0.1ml, Radix Trichosanthis Evans (trichosanthin 1mg is dissolved in the blue solution of 1ml0.5%Evans) through intravenous injection, after 30 minutes, is put to death rat, cut skull immediately, microscopically calculates the percentage rate (the results are shown in Table 4) of mast cell degranulation.
Above experimental data shows: four kinds of sprays of the present invention can suppress the rat hypertrophy cell that antigen causes and take off the granule reaction, significantly are lower than the mast cell degranulation percentage rate of normal saline matched group, and are suitable with the inhibition degree of isoproterenol; α-mannatide takes off the granule reaction to rat hypertrophy cell also significant inhibitory effect, but the inhibition degree is weaker than four kinds of sprays of the present invention slightly.Illustrate that rat hypertrophy cell that four kinds of sprays of the present invention cause antigen takes off the granule reaction curative effect is preferably arranged.
Table 4 four kinds of sprays of the present invention are to rat hypertrophy cell
Take off granule reaction influence (X+s, n=10)
Group mast cell degranulation percentage rate (%)
Normal saline matched group 95.01 ± 2.79
First kind of spray group 39.23 ± 1.69* of the present invention
Second kind of spray group 38.17 ± 1.32* of the present invention
The third spray group 37.03 ± 1.12* of the present invention
The 4th kind of spray group 36.91 ± 1.04* of the present invention
α-mannatide spray group 55.18 ± 3.45*
The different third feritin group, 38.46 ± 2.18*
Annotate: compare with the normal saline matched group: * P＜0.05
2.2.2 to infecting Sendai virus mouse red blood cell C
3The influence of formation of b garland and periphery blood T lymphocyte lipase rate of dyeing
With 1% sterilised yeast suspension of washing 3 times, filter through qualitative filter paper, recover original volume, boiled 20 minutes, fully mixing washs 2 times, makes it be the individual cells dispersity under low power lens is observed, add 37 ℃ of incubations of equivalent mice serum 15 minutes,, centrifugally be made into 5 * 10 with normal saline washing
7/ ml sensitization yeast uses liquid.Totally 160 of mices are got 20 as the blank group, and all the other every mice gives Sendai virus liquid 50 μ l from nasal cavity.Behind infective virus the 2nd day is divided into 7 groups at random: promptly normal saline matched group, four kinds of sprays of the present invention form four groups, α-mannatide spray group, levamisole group (0.195g/kg is made into 1ml solution), 20 every group.Except that the blank group, each group is corresponding administration or normal saline respectively, and volume is 1ml, divides every day and gives for three times, and administration is 7 days altogether.After the last administration 4 hours, get blood in the mouse orbit, anticoagulant heparin, the centrifuging and taking cell calculates C
3B receptor rosette rate (the results are shown in Table 5).Other gets the blood push jack, immersed in 37 ℃ of incubation liquid 30 minutes, and the water flushing, 1% peacock green dyeing, flushing is dried, and microscopy calculates T lymphocyte lipase rate of dyeing (the results are shown in Table 6).
Above experimental data shows: mouse red blood cell C behind the infection Sendai virus
3B garland formation rate significantly descends, and statistical significance is arranged; After using four kinds of sprays of the present invention, Sendai virus induced mice erythrocyte C
3B garland formation rate descends and returns to normal level again, and does not have significant difference between the blank group; α-mannatide spray and levamisole are to Sendai virus induced mice erythrocyte C
3B garland formation rate also has certain rising effect, but effect four kinds of sprays are not remarkable as described herein.Illustrate that four kinds of sprays of the present invention are to Sendai virus induced mice erythrocyte C
3The therapeutic effect that b garland formation rate descends obviously is better than single with α-mannatide spray and levamisole.
In addition, mice periphery blood T lymphocyte lipase rate of dyeing significantly descends behind the infection Sendai virus; After using four kinds of sprays of the present invention, the decline of Sendai virus induced mice periphery blood T lymphocyte lipase rate of dyeing returns to basic normal level again, compares no significant difference with the blank group; α-mannatide spray and levamisole also have certain rising effect to Sendai virus induced mice periphery blood T lymphocyte lipase rate of dyeing, but effect four kinds of sprays are not remarkable as described herein.Illustrate that the therapeutic effect that four kinds of sprays of the present invention descend to Sendai virus induced mice periphery blood T lymphocyte lipase rate of dyeing obviously is better than single with α-mannatide spray and levamisole.
Table 5 four kinds of sprays of the present invention are to infecting the Sendai virus mouse red blood cell
C
3The influence of b garland formation rate (X ± s, n=20)
Group C
3B rosette rate (%)
Blank group 11.45 ± 2.13
Normal saline matched group 7.70 ± 1.42#
First kind of spray group 11.12 ± 2.51** of the present invention
Second kind of spray group 11.23 ± 1.69** of the present invention
The third spray group 11.40 ± 1.78** of the present invention
The 4th kind of spray group 11.47 ± 1.09** of the present invention
α-mannatide spray group 8.70 ± 1.30*
Levamisole group 9.75 ± 1.60*
Annotate: compare with the normal saline matched group: * P＜0.05, * * P＜0.01; Compare #P＜0.01 with the blank group
Table 6 four kinds of sprays of the present invention are to infecting Sendai virus mice periphery blood T lymphocyte
The influence of lipase rate of dyeing (X ± s, n=20)
Group T lymphocyte lipase rate of dyeing (%)
Blank group 40.58 ± 4.37
Normal saline matched group 27.41 ± 2.25#
First kind of spray group 38.34 ± 3.26* of the present invention
Second kind of spray group 38.75 ± 3.06* of the present invention
The third spray group 39.12 ± 3.03* of the present invention
The 4th kind of spray group 40.14 ± 3.78* of the present invention
α-mannatide spray group 30.04 ± 2.29
Levamisole group 32.29 ± 2.30*
Annotate: compare with the normal saline matched group: * P＜0.05; Compare #P＜0.01 with the blank group
3 discuss
Experiment shows that four kinds of sprays of the present invention can suppress mastocyte very significantly and take off the granule reaction under antigen is attacked, and the prompting said preparation may discharge the sensitive media symptom that reduces inflammation by suppressing mastocyte.After the mouse infection Sendai virus, because the interior immune complex of serum sharply increases and/or the ability of erythrocyte removing immune complex reduces erythrocyte C
3B receptor garland formation rate obviously descends, and gives C behind four kinds of sprays of the present invention
3B receptor garland formation rate is significantly improved, and the prompting said preparation can strengthen the ability that mice antagonism Sendai virus infects.Normal development mature T lymphocyte surface of cell membrane has the lipase of certain content, and its content and function are proportionate.The T cell lipase rate of dyeing that infects the Sendai virus mice obviously descends, and this experiment prompting four kinds of sprays of the present invention have the effect that strengthens the T cell function.
The checking of the curative effect of treatment chronic pharyngitis spray of the present invention: the palace so-and-so, woman, 36 years old, the teacher, surplus main suit's pharyngalgia, dry pharynx, the pharynx foreign body sensation 10 year, once repeated multiple times was at the man hospital admissions of number, be diagnosed as chronic pharyngitis, long-term clothes antibiotics, Herba Pileae Scriptae Tabellae and other medicine, symptom repeatedly, nothing is clearly better, look into retropharyneal folliculosis, the granular lymph follicle in 6 places is arranged, the vocal cords chronic congestion, body temperature is higher to be 38.2.The patient is with this compound spray further consultation after 5 days, and subjective symptoms alleviates, and especially dry pharynx and foreign body sensation are clearly better, and checks that patient is pharyngeal, sees that pharyngeal chronic inflammatory disease alleviates, and pharynx rear wall lymph follicle reduces to 3, and vocal cords do not have hyperemia, temperature recovery to 36.8.Once more during further consultation, malaise symptoms such as conscious pharyngalgia, dry pharynx and pharynx foreign body sensation have disappeared after three weeks, and voiceless sound is hoarse, clinical examination: the pharyngeal mucosa normal color, mucosa is moistening, and the pharynx rear wall is smooth, lymph follicle disappears, and indirect laryngoscopy larynx mucomembranous color is normal.The patient is imitated very satisfied to this medication, it is simple and convenient to feel to use this medicine, no pain, and good effect has solved her and has spent a large amount of manpowers, the unsolved problem of financial resources for many years, and this medicine is pharyngitis patient's a Gospel.
The clinical efficacy statistics of treatment chronic pharyngitis spray of the present invention: 108 routine chronic pharyngitis patients are collected in this research altogether, wherein male 60 examples, women 48 examples; Age 19-62 year, the course of disease 4 months-7 years.Main performance is pharyngeal to be done, itch, hypodynia, foreign body sensation, dry cough or burn feeling.Check that the pharyngeal mucosa chronic congestion is kermesinus, retropharyneal folliculosis, lateral pharyngeal band plumpness.Throat checks no abnormal, and gets rid of stomach, backflowing property of esophagus disease.All patients use spray of the present invention and treat, every day three times, each two sprays, logotype 10 days.Curative effect determinate standard: transference cure, pharyngeal mucosa is smooth, and color and luster normally is recovery from illness; Symptom obviously alleviates, and the pharynx rear wall has congested for effective slightly; Symptom and sign all do not have obviously be improved as invalid.After treatment, recovery from illness 92 examples among the 108 routine patients account for 85.2% as a result; Effective 14 examples account for 12.9%; Invalid 2 examples account for 1.9%.The statistics total effective rate is 98.1%.
Process is collected a large amount of medication patient medical record data further investigations, concludes and sums up, and confirms that this Chinese medicine and western medicine compound spray has the General Symptomies such as heating, headache, inappetence and extremities aching pain that the elimination chronic pharyngitis causes.To pharyngeal sense of discomfort, as dry, itch, scorching hot, hypodynia and foreign body sensation etc. are evident in efficacy.Pharyngeal mucous hyperemia after the medication, thicken phenomenon and fade away, the rheuminess thing reduces also and disappears.
Claims (9)
1, a kind of spray for the treatment of chronic pharyngitis is characterized in that: it mainly includes the Chinese medicine 60-90% of α-mannatide 10-40%, antiinflammatory antipyretic-antalgic class.
2, a kind of spray for the treatment of chronic pharyngitis according to claim 1 is characterized in that: described composition includes the Chinese medicine 10-50% of α-mannatide 10-40%, antiinflammatory antipyretic-antalgic class, the Chinese medicine 10-60% of antitussive class.
3, a kind of spray for the treatment of chronic pharyngitis according to claim 1 is characterized in that: described composition includes Chinese medicine 10-40%, the Chinese medicine 10-50% of antitussive class, the Chinese medicine 10-60% of reduction capillary permeability of α-mannatide 10-30%, antiinflammatory antipyretic-antalgic class.
4, a kind of spray for the treatment of chronic pharyngitis according to claim 1 is characterized in that: described composition include Chinese medicine 10-35%, the antitussive class of α-mannatide 10-25%, antiinflammatory antipyretic-antalgic class Chinese medicine 10-40%, reduce capillary permeability Chinese medicine 10-50%, reduce blood viscosity, improve the Chinese medicine 10-40% of hemorheological property.
5, a kind of spray for the treatment of chronic pharyngitis according to claim 1 is characterized in that: the Chinese medicine of described antiinflammatory antipyretic-antalgic class: Flos Lonicerae, Radix Isatidis, Fructus Forsythiae, Folium Isatidis, Indigo Naturalis, Rhizoma Coptidis, Radix Scutellariae, Cortex Phellodendri, Caulis Fibraureae, Herba Andrographis, Herba Violae, Herba Taraxaci, Herba Patriniae, Rhizoma Paridis, Radix Gentianae, Radix Sophorae Tonkinensis, the Rhizoma Anemarrhenae, Cortex Magnoliae Officinalis, Cortex Moutan, the Radix Paeoniae Alba, Spica Prunellae, Fructus Trichosanthis, Calculus Bovis, Bulbus Allii, Fructus Chebulae; The Chinese medicine of antitussive class: Radix Rehmanniae, Radix Glycyrrhizae, Bulbus Fritillariae Thunbergii, Fritillary flower, Folium Eriobotryae, Folium Rhododendri Daurici, Folium et Cacumen Rhododendri Mariae, Herba Cymbopogonis, the Rhizoma Pinelliae, Radix Peucedani, Cortex Mori, Fructus Aristolochiae, the Rhizoma Anemarrhenae, Semen Plantaginis, Bulbus Lilii, Radix Ophiopogonis, Rhizoma Polygoni Cuspidati; Reduce the Chinese medicine of capillary permeability: the Flos Sophorae Immaturus, Flos Sophorae, Fructus Forsythiae, Rhizoma Imperatae, the Radix Astragali, Caulis Sargentodoxae, Radix Scutellariae, Cornu Bubali, CORTEX ACANTHOPANACIS, Pericarpium Citri Reticulatae Viride, Pericarpium Citri Reticulatae; Reduce blood viscosity, improve the Chinese medicine of hemorheological property: ligustrazine, Radix Salviae Miltiorrhizae, the Radix Astragali, Radix Rehmanniae, Hirudo, Ganoderma, Radix Puerariae, Flos Carthami, Radix Paeoniae Rubra.
6, a kind of spray for the treatment of chronic pharyngitis according to claim 1, it is characterized in that: the composition of the every 1000g of described medicament is; α-mannatide 400g, Flos Lonicerae 40g, Radix Isatidis 50g, Fructus Forsythiae 20g, Folium Isatidis 40g, Indigo Naturalis 20g, Rhizoma Coptidis 20g, Radix Scutellariae 20g, Cortex Phellodendri 20g, Caulis Fibraureae 20g, Herba Andrographis 30g, Herba Violae 30g, Herba Taraxaci 40g, Herba Patriniae 10g, Rhizoma Paridis 10g, Radix Gentianae 10g, Radix Sophorae Tonkinensis 30g, Rhizoma Anemarrhenae 20g, Cortex Magnoliae Officinalis 10g, Cortex Moutan 40g, Radix Paeoniae Alba 20g, Spica Prunellae 10g, Fructus Trichosanthis 10g, Calculus Bovis 10g, Bulbus Allii 50g, Fructus Chebulae 20g.
7, a kind of spray for the treatment of chronic pharyngitis according to claim 1, it is characterized in that: the composition of the every 1000g of described medicament is; α-mannatide 365g, Flos Lonicerae 20g; Radix Isatidis 10g; Fructus Forsythiae 20g; Folium Isatidis 15g; Indigo Naturalis 20g; Rhizoma Coptidis 10g; Radix Scutellariae 15g; Cortex Phellodendri 20g; Caulis Fibraureae 15g; Herba Andrographis 20g; Herba Violae 15g; Herba Taraxaci 25g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 20g; Rhizoma Anemarrhenae 15g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 25g; Radix Paeoniae Alba 20g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 30g; Fructus Chebulae 20g; Radix Rehmanniae 10g; Radix Glycyrrhizae 20g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 25g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g.
8, a kind of spray for the treatment of chronic pharyngitis according to claim 1, it is characterized in that: the composition of the every 1000g of described medicament is; α-mannatide 250g, Flos Lonicerae 20g; Radix Isatidis 20g; Folium Isatidis 15g; Indigo Naturalis 10g; Rhizoma Coptidis 10g; Cortex Phellodendri 10g; Caulis Fibraureae 15g; Herba Andrographis 10g; Herba Violae 15g; Herba Taraxaci 15g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 15g; Rhizoma Anemarrhenae 15g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 25g; Radix Paeoniae Alba 10g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 20g; Fructus Chebulae 15g; Radix Rehmanniae 10g; Radix Glycyrrhizae 20g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 25g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g; Flos Sophorae Immaturus 20g; Flos Sophorae 20g; Fructus Forsythiae 30g; Rhizoma Imperatae 20g; Radix Astragali 15g; Caulis Sargentodoxae 20g; Radix Scutellariae 20g; Cornu Bubali 20g; CORTEX ACANTHOPANACIS 20g; Pericarpium Citri Reticulatae Viride 10g; Pericarpium Citri Reticulatae 15g.
9, a kind of spray for the treatment of chronic pharyngitis according to claim 1, it is characterized in that: the composition of the every 1000g of described medicament is; α-mannatide 200g, Flos Lonicerae 20g; Radix Isatidis 10g; Folium Isatidis 15g; Indigo Naturalis 10g; Rhizoma Coptidis 10g; Cortex Phellodendri 10g; Caulis Fibraureae 15g; Herba Andrographis 10g; Herba Violae 10g; Herba Taraxaci 15g; Herba Patriniae 10g; Rhizoma Paridis 10g; Radix Gentianae 10g; Radix Sophorae Tonkinensis 15g; Rhizoma Anemarrhenae 10g; Cortex Magnoliae Officinalis 10g; Cortex Moutan 15g; Radix Paeoniae Alba 10g; Spica Prunellae 10g; Fructus Trichosanthis 10g; Calculus Bovis 10g; Bulbus Allii 15g; Fructus Chebulae 15g; Radix Rehmanniae 10g; Radix Glycyrrhizae 15g; Bulbus Fritillariae Thunbergii 10g; Fritillary flower 5g; Folium Eriobotryae 10g; Folium Rhododendri Daurici 20g; Folium et Cacumen Rhododendri Mariae 15g; Herba Cymbopogonis 20g; Rhizoma Pinelliae 10g; Radix Peucedani 10g; Cortex Mori 10g; Fructus Aristolochiae 10g; Rhizoma Anemarrhenae 15g; Semen Plantaginis 15g; Bulbus Lilii 20g; Radix Ophiopogonis 10g; Rhizoma Polygoni Cuspidati 10g; Flos Sophorae Immaturus 20g; Flos Sophorae 20g; Fructus Forsythiae 30g; Rhizoma Imperatae 20g; Caulis Sargentodoxae 20g; Radix Scutellariae 20g; Cornu Bubali 20g; CORTEX ACANTHOPANACIS 20g; Pericarpium Citri Reticulatae Viride 10g; Pericarpium Citri Reticulatae 15g; ligustrazine 10g; Radix Salviae Miltiorrhizae 10g; Radix Astragali 30g; Radix Rehmanniae 10g; Hirudo 10g; Ganoderma 10g; Radix Puerariae 15g; Flos Carthami 10g; Radix Paeoniae Rubra 10g.
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| CN200410104428A CN100592919C (en) | 2003-12-31 | 2004-12-19 | Spray for treating chronic pharyngitis |
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| CN200310122249.1 | 2003-12-31 | ||
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| CN100592919C CN100592919C (en) | 2010-03-03 |
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| CN101850073A (en) * | 2010-06-08 | 2010-10-06 | 幸泽刚 | Chinese medicinal composition for treating chronic pharyngitis |
| CN101485827B (en) * | 2009-02-17 | 2011-03-23 | 张跃 | Medicament for treating chronic inflammation disease of throat |
| CN102357155A (en) * | 2011-10-27 | 2012-02-22 | 潘根起 | Traditional Chinese medicine combination for treating herpes zoster |
| CN102671075A (en) * | 2012-05-09 | 2012-09-19 | 太仓市伟基生物科技有限公司 | Chewing gum for treating chronic pharyngitis |
| CN103566182A (en) * | 2013-10-24 | 2014-02-12 | 宋海平 | Traditional Chinese medicine preparation for preventing and treating radioactive mucositis |
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| CN104922346A (en) * | 2015-04-29 | 2015-09-23 | 金先琦 | Traditional Chinese medicine for treating chronic pharyngitis |
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| CN101850073B (en) * | 2010-06-08 | 2011-11-09 | 幸泽刚 | Chinese medicinal composition for treating chronic pharyngitis |
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| CN102357155A (en) * | 2011-10-27 | 2012-02-22 | 潘根起 | Traditional Chinese medicine combination for treating herpes zoster |
| CN102357155B (en) * | 2011-10-27 | 2012-11-14 | 潘根起 | Traditional Chinese medicine combination for treating herpes zoster |
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| CN102671075B (en) * | 2012-05-09 | 2013-11-27 | 太仓市伟基生物科技有限公司 | Chewing gum for treating chronic pharyngitis |
| CN103566141B (en) * | 2012-10-31 | 2015-11-25 | 张绪伟 | A kind of preparation method for the treatment of the medicament of chronic pharyngitis |
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| CN103566182A (en) * | 2013-10-24 | 2014-02-12 | 宋海平 | Traditional Chinese medicine preparation for preventing and treating radioactive mucositis |
| CN103784709A (en) * | 2014-01-28 | 2014-05-14 | 刘永秋 | Chinese medicinal composition for treating chronic pharyngitis and preparation method thereof |
| CN104027413A (en) * | 2014-06-18 | 2014-09-10 | 潘井生 | Traditional Chinese medicine composition for treating dental ulcer |
| CN104189572A (en) * | 2014-08-22 | 2014-12-10 | 郎社芳 | Traditional Chinese medicine composition for treating chronic pharyngitis, as well as aerosol and preparation thereof |
| CN104922346A (en) * | 2015-04-29 | 2015-09-23 | 金先琦 | Traditional Chinese medicine for treating chronic pharyngitis |
| CN104887936A (en) * | 2015-05-28 | 2015-09-09 | 陈有伦 | Chinese medicinal composition for treating swelling and pain in throat |
| CN106266854A (en) * | 2015-06-02 | 2017-01-04 | 中南林业科技大学 | A kind of treat acute/chronic pharyngitis Chinese medicine preparation |
| CN105194249A (en) * | 2015-09-18 | 2015-12-30 | 马鞍山德宏堂生物科技有限公司 | Traditional Chinese medicine decoction for treating sphagitis |
| CN105920295A (en) * | 2016-04-28 | 2016-09-07 | 浙江医药高等专科学校 | Traditional Chinese medicine composition with effects of moistening throat and relieving sore throat and its preparation method and use |
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| CN107929710A (en) * | 2018-01-27 | 2018-04-20 | 朱增伟 | For treating the Chinese medicine composition of chronic pharyngitis |
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| CN100592919C (en) | 2010-03-03 |
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