CN100522183C - Earthworm acidic-part medicine for treating cough asthma disease and preparing method - Google Patents
Earthworm acidic-part medicine for treating cough asthma disease and preparing method Download PDFInfo
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- CN100522183C CN100522183C CNB2006100242720A CN200610024272A CN100522183C CN 100522183 C CN100522183 C CN 100522183C CN B2006100242720 A CNB2006100242720 A CN B2006100242720A CN 200610024272 A CN200610024272 A CN 200610024272A CN 100522183 C CN100522183 C CN 100522183C
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Abstract
The present invention relates to an earthworm acid region medicine for curing cough and asthma diseases and its preparation method. Said earthworm acid region medicine contains fatty acid, amino acid and purine compound, and can be used for curing the diseases of asthma and chronic bronchitis, etc. Its preparation method includes the following steps: using strong base anion-exchange resin to adsorb aqueous extract of earthworm, then using dilute hydrochloric acid or alcohol-containing dilute hydrochloric acid to make elution.
Description
Technical field
The present invention relates to medicine of a kind of pharmaceutical engineering technical field and preparation method thereof, specifically, what relate to is a kind of earthworm acidic-part medicine for the treatment of cough illness and preparation method thereof.
Background technology
Asthma is that a kind of what caused by multiple factor is the immune allergic disorder of feature with reversibility airway obstruction, airway hyperreactivity and airway inflammation.Chronic bronchitis (abbreviation chronic bronchitis) is the chronic nonspecific inflammation of trachea, bronchial mucosa and surrounding tissue.Clinical manifestation such as chronic bronchitis, obstructive disease of lung primary symptom is " cough, expectorant, breathe heavily ", and doctor's custom abbreviates the chronic cough and asthma disease as.Cough illness is the commonly encountered diseases in serious harm people health Jiankang, frequently-occurring disease.Cough illness does not still have specific Therapeutic Method at present, and in China, clinical experience for many years shows that this disease of Chinese medicine has higher curative effect, and toxic and side effects is little.The Chinese medicine active substance of research determined curative effect, safe and reliable anti-cough illness, further developing new drug will be benefited the patient, produce remarkable social benefit and economic benefit.
" Pheretima recorded of Chinese pharmacopoeia version in 2005 is the dry body of huge earthworm animal Pheretima aspergillum Pheretimaaspergillum of section (E.Perrier), popular Pheretimatschiliensis Pheretima vulagris Chen, William Pheretimatschiliensis Pheretima guillelm (Michaelsen) or the blind Pheretimatschiliensis Pheretima of comb pectiniferaMichaelsen, have heat clearing away arresting convulsion, collateral dredging, relieving asthma, the effect of diuresis, it is a kind of conventional animal medical material, pharmacologically active is remarkable, aboundresources.The component of bibliographical information Pheretima can block histamine receptor, and antihistamine is made tracheospasm and increases capillary permeability.Pheretima 90% alcohol extract can obviously increase rat and rabbit trachea lung perfusion flow, and the bronchoconstriction that can cause antihistamine and pilocarpine.Fresh Pheretima body fluid is extremely sensitive to the common pathogenic bacteria of pulmonary infection.Pheretima can promote immunocyte significantly---the Turnover of Mouse Peritoneal Macrophages activation has immunoregulation effect.
Do not see the report of acidic site of Pheretima about cough illness such as treatment asthma, chronic bronchitiss and preparation method thereof in existing document and the concrete practice as yet.Pheretima is direct as the prescription raw material with the medicated powder form generally speaking, perhaps pulverizes the back and decocts extracting juice jointly with other flavour of a drug entry, perhaps carries out water extract-alcohol precipitation simply.Prescribed preparation of including of 2005 editions Chinese Pharmacopoeias " lung-clearing anti-inflammatory ball " for example promptly is that Pheretima and Herba Ephedrae, Semen Armeniacae Amarum (stir-fry), Fructus Arctii, Gypsum Fibrosum, Semen Lepidii (Semen Descurainiae) are ground into fine powder, with Cornu Saigae Tataricae impalpable powder and artificial Calculus Bovis's powder facing-up, makes water-honeyed pill again.For example Lee like really the silver-colored imperial DINGCHUAN TANG of " silver-colored imperial DINGCHUAN TANG treatment childhood asthma 72 examples " (2002 the 23rd the 6th phases of volume of the Shaanxi traditional Chinese medical science) report be Pheretima with Semen Ginkgo, Herba Ephedrae (processed), system Semen Armeniacae Amarum, Cortex Lycii, honey-fried CORTEX MORI, Fructus Perillae, Radix Stephaniae Tetrandrae, Radix Angelicae Sinensis, Radix Paeoniae Rubra the totally 10 medical material mixing waters of distinguishing the flavor of take juice.For example record the sterile water solution after the earthworm injection of in 1987 version " Guangdong Province's drug standard " first volume filters for the Pheretima water extract-alcohol precipitation again.In the Pheretima application form, through separating remove impurity, effective site that Pheretima is relievingd asthma and invalid target do not mix utilization for Pheretima powder or Pheretima extracting solution, and active ingredient chemical substance basis is unclear, be difficult to set up the quality standard of medicine, be difficult to effectively control drug quality.
Summary of the invention
The present invention is directed to the deficiencies in the prior art, a kind of earthworm acidic-part medicine that is used for the treatment of cough illness and preparation method thereof is provided.Make its foundation that helps drug standard and quality control.
The present invention is achieved by the following technical solutions, the earthworm acidic-part medicine of treatment cough illness disclosed by the invention is the effective site of extracting from Pheretima that contains fatty acid, aminoacid, purine three compounds, wherein aminoacid and purine mass fraction sum are 50%～80% and the material that made by following operation: get dried Pheretima, be ground into coarse powder, add for the first time 2～8 times of water gaging soaking at room temperature of Pheretima, 2～5h, filter; Add for the second time 40 ℃～50 ℃ warm macerating 1～3h of 2～6 times of water gagings of Pheretima, filter, repeat this operation once, filter, merge three times filtrate, low-speed centrifugal 3～15min gets centrifuged supernatant, is evaporated to 2～6 times of volumes of Pheretima consumption, adding 95% ethanol again makes and contains alcohol amount and reach 70%, leave standstill 8～24h, centrifugal, get supernatant, supernatant is evaporated to 2～5 times of volumes of Pheretima consumption, again it is passed through strong-base anion-exchange resin, blade diameter length ratio 1:10, resin demand are 2～5 times of Pheretima; It is neutral colourless earlier being eluted to eluent with the deionized water of 3～15 times of column volumes, and reuse contains 0%～35% alcoholic solution eluting of 0.1N hydrochloric acid, collects eluent, and 40 ℃～50 ℃ drying under reduced pressure promptly obtain being used for the treatment of the acid effect of the Pheretima position medicine of cough illness.
Described fatty acid comprises: oxalic acid, succinic acid, malic acid, adipic acid, 1,3-propanedicarboxylic acid, 3-hydroxycaproic acid, Palmic acid, lauric acid,-alkene hexadecanoic acid, tridecanoic acid, myristic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, linoleic acid, oleic acid, stearic acid, arachidic acid, eicosatrienoic acid, arachidonic acid, the pentaene arachic acid, the triolefin arachic acid, six alkene behenic acids, 12-methyl alkanoic acid, 12-methyl tetradecanoic acid, 14-methyl pentadecanoic acid, 14-methyl hexadecanoic acid, nonadecane olefin(e) acid etc.
Described aminoacid comprises: aspartic acid, isoleucine, threonine, leucine, serine, tyrosine, glutamic acid, phenylalanine, proline, γ-An Jidingsuan, glycine, ornithine, alanine, lysine, cystine, histidine, valine, tryptophan, methionine, arginine etc.
Described purine comprises xanthine, hypoxanthine etc.
The present invention is above-mentioned, and to be used for the treatment of the preparation method of earthworm acidic-part medicine of cough illness as follows: get dried Pheretima, be ground into coarse powder, add 2～8 times of water gaging soaking at room temperature of Pheretima, 2～5h for the first time, filter; Add for the second time 40 ℃～50 ℃ warm macerating 1～3h of 2～6 times of water gagings of Pheretima, filter, repeat this operation once, filter, merge three times filtrate, low-speed centrifugal 3～15min gets centrifuged supernatant, is evaporated to 2～6 times of volumes of Pheretima consumption, adding 95% ethanol again makes and contains alcohol amount and reach 70%, leave standstill 8～24h, centrifugal, get supernatant, supernatant is evaporated to 2～5 times of volumes of Pheretima consumption, again it is passed through strong-base anion-exchange resin, resin column blade diameter length ratio 1:10, resin demand are 2～5 times of Pheretima; It is neutral colourless earlier being eluted to eluent with the deionized water of 3～15 times of column volumes, and reuse contains 0%～35% alcoholic solution eluting of 0.1N hydrochloric acid, collects eluent, and 40 ℃～50 ℃ drying under reduced pressure promptly obtain being used for the treatment of the acid effect of the Pheretima position medicine of cough illness.
Described strong-base anion-exchange resin can be the strong-base anion-exchange resin of HZ-201 or other different models.
Ninhydrin colorimetry is measured the content of total amino acids in the acid sites, and the HPLC method is measured the content of purine in the acid sites.
Earthworm acidic-part medicine of the present invention is compared with the Pheretima raw medicinal herbs, has following advantage: 1. through the extraction separation remove impurity, removed for the cough illness inert matter, the antiasthmatic activity component content is improved greatly.2. acidic site of Pheretima does not have the intrinsic fishy smell of Pheretima, uses its preparation of making and more helps patient's acceptance.
In a word, with existing Pheretima powder or the Pheretima decocting liquid phase ratio used, earthworm acidic-part medicine chemical composition of the present invention is clear, pharmacologically active strengthens, and has reduced drug dose, more convenient patient's medication, increase the compliance of patient's medication, simultaneously, help the foundation and the quality control of drug standard, making the application Pheretima be prepared into various anti-cough illness preparations becomes possibility.
Description of drawings
The extraction separation flow chart at Fig. 1 Pheretima three positions
The diastole effect that the guinea-pig isolated tracheal smooth muscle that Fig. 2 Pheretima three positions cause 10 μ M carbachols shrinks
The diastole effect that the guinea-pig isolated tracheal smooth muscle that Fig. 3 Pheretima three positions cause 10 μ M group ammonia shrinks
Fig. 4 Pheretima three positions cause the inhibitory action that rat tracheal mucosa short circuit current rises to 10 μ M carbachols
Fig. 5 acidic site of Pheretima GC-MS total ion current collection of illustrative plates
The HPLC collection of illustrative plates of Fig. 6 acidic site of Pheretima
The specific embodiment
The present invention is described in detail below in conjunction with embodiment and accompanying drawing.
The embodiment 1 screening Pheretima effective site of relievining asthma
3 positions that prepare Pheretima respectively comprise acidic site of Pheretima, Pheretima alkalescence position, Pheretima water extract-alcohol precipitation position (preparation process is seen Fig. 1).Observe the relexation that guinea pig tracheal smooth muscle tension force that three extract parts cause carbachol (CCh) and histamine (His) increases subsequently, and the inhibitory action (suppressing the sputum secretory action) that the rat tracheal epithelium short circuit current that CCh causes is increased.3 Pheretima extract parts are seen Fig. 2 and Fig. 3 to the diastole effect of tracheal smooth muscle, the result show diastole effect that guinea-pig isolated tracheal smooth muscle that 3 kinds of Pheretima extract parts cause for 10 μ M carbachols shrinks all a little less than, acidic site of Pheretima has maximum diastolic rate, is 35.53%; Excellent sub-Pheretima alkalescence position, the equal significance of diastole effect ground and water extract-alcohol precipitation position that the guinea-pig isolated tracheal smooth muscle that acidic site of Pheretima causes for 10 μ M group ammonia shrinks, its maximum diastolic rate reaches 77.99%, illustrate in the Pheretima that the effective site that tracheal smooth muscle is played the diastole effect is acid sites, its effect mainly is the contraction that anti-group ammonia causes.
3 kinds of Pheretima positions cause the decline effect that short circuit current rises to 10 μ M carbachols, see Fig. 4, the result shows that the decline effect that short circuit current that acidic site of Pheretima causes 10 μ M carbachols rises is better than Pheretima alkalescence position and water extract-alcohol precipitation position, illustrate that the effective site of Pheretima inhibition tracheal mucosa ion secretion is acid sites.
In sum, in three extraction separation positions of Pheretima, acidic site of Pheretima has the diastole effect of the guinea-pig isolated tracheal smooth muscle contraction that causes for group ammonia.Simultaneously, acidic site of Pheretima suppresses the effect of isolated rat tracheal mucosa ion secretion in addition, thereby can suppress excessive sputum secretion.In a word, the Pheretima effective site that is used for cough illness treatment is acidic site of Pheretima.
Embodiment 2 preparation acidic site of Pheretima
Preparation method:
Get dried Pheretima 1000g, be ground into coarse powder, add 5 times of water gaging soaking at room temperature of Pheretima 3h for the first time, filter; Add for the second time 45 ℃ of warm macerating 2h of 4 times of water gagings of Pheretima, filter, repeat this operation once, filter, merge three times filtrate, low-speed centrifugal 9min gets centrifuged supernatant, be evaporated to 4 times of volumes of Pheretima consumption, add 95% ethanol again and make and contain alcohol amount and reach 70%, leave standstill 16h, centrifugal, get supernatant, supernatant is evaporated to 3 times of volumes of Pheretima consumption.Again it is passed through strong-base anion-exchange resin (resin column blade diameter length ratio 1:10, resin demand are 3 times of Pheretima).It is neutral colourless earlier being eluted to eluent with the deionized water of 9 times of column volumes, and reuse contains 20% alcoholic solution eluting of 0.1N hydrochloric acid, collection yellow eluent part, promptly the obtain medical treatment earthworm acidic-part medicine of cough illness of 45 ℃ of drying under reduced pressure.
Implementation result:
The percentage by weight that amino acid content accounts for whole acid sites is 64.68%, and purine content accounts for 0.68%.
Embodiment 3 preparation acidic site of Pheretima
Preparation method:
Get dried Pheretima 1000g, be ground into coarse powder, add 2 times of water gaging soaking at room temperature of Pheretima 2h for the first time, filter; Add for the second time 40 ℃ of warm macerating 1h of 2 times of water gagings of Pheretima, filter, repeat this operation once, filter, merge three times filtrate, low-speed centrifugal 3min gets centrifuged supernatant, be evaporated to 2 times of volumes of Pheretima consumption, add 95% ethanol again and make and contain alcohol amount and reach 70%, leave standstill 8h, centrifugal, get supernatant, supernatant is evaporated to 2 times of volumes of Pheretima consumption.Again it is passed through strong-base anion-exchange resin (resin column blade diameter length ratio 1:10, resin demand are 2 times of Pheretima).It is neutral colourless earlier being eluted to eluent with the deionized water of 3 times of column volumes, reuse 0.1N hydrochloric acid solution eluting, collection yellow eluent part, promptly the obtain medical treatment earthworm acidic-part medicine of cough illness of 50 ℃ of drying under reduced pressure.
Implementation result: the percentage by weight that aminoacid accounts for whole acid sites is 50.36%, and purine content accounts for 0.44%.
Embodiment 4 preparation acidic site of Pheretima
Preparation method:
Get dried Pheretima 1000g, be ground into coarse powder, add 8 times of water gaging soaking at room temperature of Pheretima 5h for the first time, filter; Add for the second time 50 ℃ of warm macerating 3h of 6 times of water gagings of Pheretima, filter, repeat this operation once, filter, merge three times filtrate, low-speed centrifugal 15min gets centrifuged supernatant, be evaporated to 6 times of volumes of Pheretima consumption, add 95% ethanol again and make and contain alcohol amount and reach 70%, leave standstill 24h, centrifugal, get supernatant, supernatant is evaporated to 5 times of volumes of Pheretima consumption.Again it is passed through strong-base anion-exchange resin (resin column blade diameter length ratio 1:10, resin demand are 5 times of Pheretima).It is neutral colourless earlier being eluted to eluent with the deionized water of 15 times of column volumes, and reuse contains 35% alcoholic solution eluting of 0.1N hydrochloric acid, collection yellow eluent part, promptly the obtain medical treatment earthworm acidic-part medicine of cough illness of 40 ℃ of drying under reduced pressure.
Implementation result: the percentage by weight that aminoacid accounts for whole acid sites is 78.45%, and purine content accounts for 1.25%.
Each composition of 5 pairs of acidic site of Pheretima of embodiment is confirmed
With the ethyl chloroformate ethyl esterization, utilize gas chromatography/mass spectrometry/computer multiple techniques to study its composition embodiment 2 gained acidic site of Pheretima, each component is retrieved by computer spectrum storehouse, and list of references confirmed chemical compound, the results are shown in Figure 5.
Wherein each peak is represented respectively: appearance time 6.21min is a Fumaric acid, and 6.31min is a succinic acid, and 7.68min is a malic acid, 7.71min be alanine, 7.91min is a glycine, 9.25min is an adipic acid, 9.4min be valine, 10.65min is a leucine, 10.70min is a 1,3-propanedicarboxylic acid, 10.89min be isoleucine, 11.25min be aspartic acid, 12.66min is the 3-hydroxycaproic acid, 14.20min is a glutamic acid, 17.34min be myristic acid, 18.73min be phenylalanine, 18.85min is a proline, 21.40min is an alkene hexadecanoic acid, 21.76 be lysine, 21.92min be hexadecanoic acid, 23.90min is a heptadecanoic acid, 24.75min is a linoleic acid, 24.81min be oleic acid, 25.08min be stearic acid, 26.17min is an arachidonic acid, 26.22min is the pentaene arachic acid, 26.32min be the triolefin arachic acid, 26.7min is a tyrosine, 27.40min is six alkene behenic acids, 27.49min be tryptophan, 27.69min is a cysteine.
Embodiment 2 gained acidic site of Pheretima are carried out HPLC analyze, by comparing with reference substance, result: 5.316min is a hypoxanthine, and 5.71min is an xanthine, and 9.898min is the tyrosine (see figure 6).
The research of embodiment 6 pharmacological toxicologies
1. antiasthmatic effect
Embodiment 2 gained acidic site of Pheretima 0.3～0.6mg/ml causes that to 10uM group ammonia the guinea pig trachea ring contraction of exsomatizing has tangible diastole effect.Press 360mg, 720mg, 1080mg/kg are to the Cavia porcellus gastric infusion, and the allergic asthma that egg protein is caused obviously prolongs incubation period, to sucking asthma also obviously prolongation incubation period that histamine causes.
2. to the influence of the phenol red excretion amount of mice trachea
Embodiment 2 gained acidic site of Pheretima are pressed 180mg, and 360mg, 540mg/kg can prolong the mouse cough incubation period that strong aqua ammonia and sulfur dioxide bring out to the mouse stomach administration, reduce the cough number of times, and the phenol red excretion amount of mice trachea is obviously increased.
3. antiinflammatory action
Embodiment 2 gained acidic site of Pheretima can suppress the growth of staphylococcus aureus, Diplococcus pneumoniae, alpha streptococcus and to the swelling of anti-inflammatory Mus ear.Relatively there were significant differences with the matched group gentamycin; Can reduce the quantity of skin exfoliative cyte and inflammatory cell in patient's sputum, prompting has antiinflammatory action.
4. enhance immunity effect
Select for use the activation rate of macrophage to estimate immunization, the result shows, embodiment 2 gained acidic site of Pheretima have and promote immunocyte significantly---the effect of macrophage activation, concentration is comparatively remarkable effect in 0.45%～0.9% o'clock.Too high or too low effect is more weak or do not have an effect.
5. acute toxicity test
Use the Wistar healthy mice, adopt oral and two kinds of administering modes of tail vein injection, measure the median lethal dose(LD 50) (LD50) of embodiment 2 gained acidic site of Pheretima, oral is 1753mg/Kg ± 104mg/Kg, and lumbar injection is 985mg/Kg ± 87mg/Kg.
Above-mentioned evidence acidic site of Pheretima safety of the present invention, toxicity are low, have antitussive, eliminate the phlegm, relieving asthma, the effect of cough illness patient's immunity of organisms such as antiinflammatory, enhancing asthma, chronic bronchitis.
Claims (6)
1, a kind of earthworm acidic-part medicine for the treatment of cough illness, it is characterized in that, contain aminoacid, fatty acid, purine, wherein aminoacid and purine mass fraction sum are 50%～80% and the material that made by following operation: get dried Pheretima, be ground into coarse powder, add for the first time 5～8 times of water gaging soaking at room temperature of Pheretima, 3～5h, filter; Add for the second time 40 ℃～50 ℃ warm macerating 1～3h of 3～6 times of water gagings of Pheretima, filter, repeat this operation once, filter, merge three times filtrate, low-speed centrifugal 5～10min gets centrifuged supernatant, is evaporated to 3～6 times of volumes of Pheretima consumption, adding 95% ethanol again makes and contains alcohol amount and reach 70%, leave standstill 12～24h, centrifugal, get supernatant, supernatant is evaporated to 3～5 times of volumes of Pheretima consumption, again its flow velocity with 3～5ml/min is passed through strong-base anion-exchange resin, blade diameter length ratio 1:10, resin demand are 2～3 times of Pheretima; It is neutral colourless that first deionized water with 5～15 times of column volumes is eluted to eluent, it is that 0%～35% 0.1N hydrochloric acid solution is with 3～5ml/min eluting that reuse contains concentration of alcohol, collect eluent, 40 ℃～50 ℃ drying under reduced pressure promptly obtain being used for the treatment of the acid position medicine of imitating of Pheretima of cough illness.
2, the earthworm acidic-part medicine of treatment cough illness according to claim 1 is characterized in that, described fatty acid comprises: oxalic acid, succinic acid, malic acid, adipic acid, 1,3-propanedicarboxylic acid, 3-hydroxycaproic acid, Palmic acid, lauric acid, one alkene hexadecanoic acid, tridecanoic acid, myristic acid, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, linoleic acid, oleic acid, stearic acid, arachidic acid, eicosatrienoic acid, arachidonic acid, the pentaene arachic acid, the triolefin arachic acid, six alkene behenic acids, 12-methyl alkanoic acid, 12-methyl tetradecanoic acid, 14-methyl pentadecanoic acid, 14-methyl hexadecanoic acid, the nonadecane olefin(e) acid.
3, the earthworm acidic-part medicine of treatment cough illness according to claim 1, it is characterized in that described aminoacid comprises: aspartic acid, isoleucine, threonine, leucine, serine, tyrosine, glutamic acid, phenylalanine, proline, γ-An Jidingsuan, glycine, ornithine, alanine, lysine, cystine, histidine, valine, tryptophan, methionine, arginine.
4, the earthworm acidic-part medicine of treatment cough illness according to claim 1 is characterized in that, described purine comprises xanthine, hypoxanthine.
5, a kind of preparation method for the treatment of the earthworm acidic-part medicine of cough illness is characterized in that, gets dried Pheretima, is ground into coarse powder, adds 5～8 times of water gaging soaking at room temperature of Pheretima, 3～5h for the first time, filters; Add for the second time 40 ℃～50 ℃ warm macerating 1～3h of 3～6 times of water gagings of Pheretima, filter, repeat this operation once, filter, merge three times filtrate, low-speed centrifugal 5～10min, get centrifuged supernatant, be evaporated to 3～6 times of volumes of Pheretima consumption, adding 95% ethanol again makes and contains alcohol amount and reach 70%, leave standstill 12～24h, centrifugal, get supernatant, supernatant is evaporated to 3～5 times of volumes of Pheretima consumption, again its flow velocity with 3～5ml/min is passed through strong-base anion-exchange resin, blade diameter length ratio 1:10, resin demand are 2～3 times of Pheretima, it is neutral colourless that first deionized water with 5～10 times of column volumes is eluted to eluent, reuse contain concentration of alcohol be 25%～35% 0.1N hydrochloric acid solution with 3～5ml/min eluting, collect the yellow eluent part, 40 ℃～50 ℃ drying under reduced pressure promptly obtain being used for the treatment of the acid position medicine of imitating of Pheretima of cough illness.
6, the preparation method of the earthworm acidic-part medicine of treatment cough illness according to claim 5 is characterized in that, described resin is a strong-base anion-exchange resin.
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| BR112013006044B1 (en) | 2010-09-24 | 2022-03-29 | University Of Florida Research Foundation, Inc | Therapeutic composition for enteral administration comprising serine and valine as free amino acids, as well as packaging containing said composition |
| AU2014249078B2 (en) * | 2013-03-11 | 2018-07-26 | University Of Florida Research Foundation, Incorporated | Materials and methods for improving lung function and for prevention and/or treatment of radiation-induced lung complications |
| CN106265750A (en) * | 2015-06-10 | 2017-01-04 | 葛以东 | A kind of formula controlling childhood asthma |
| CN110592052B (en) * | 2019-09-22 | 2020-09-29 | 内蒙古新宏生物科技有限公司 | Production method and application of lumbruse polypeptide antibacterial nutrient solution |
| EP4084784A4 (en) * | 2020-01-13 | 2024-01-17 | Univ Monash | Compositions and methods |
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| 正交试验法优选广地龙提取工艺. 高华宏，刘昌，王文琼，潘玲.湖北中医学院学报，第6卷第3期. 2004 |
| 正交试验法优选广地龙提取工艺. 高华宏，刘昌，王文琼，潘玲.湖北中医学院学报，第6卷第3期. 2004 * |
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