CN113491760A - Medicine for treating respiratory system diseases - Google Patents

Abstract

The invention relates to a medicament for treating respiratory diseases, which is used for treating or preventing pulmonary embolism, or interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, lung diseases caused by smoking and complicated hemorrhage.

Description

Medicine for treating respiratory system diseases

Technical Field

The invention relates to the field of medicine, in particular to a medicine for treating respiratory system diseases.

Background

Protein is an important component for composing all cells and tissues of human body. All important components of the body require the involvement of proteins. Protein accounts for about 18% of the total mass of the human body, and most importantly, it is related to life phenomena. The protein is a substance with a certain spatial structure formed by the way that polypeptide chains consisting of amino acids in a dehydration condensation mode are folded in a coiling way. After the translational synthesis of protein, glycosylation is a main posttranslational modification form of protein, and some amino acid residues can be subjected to posttranslational modification to generate chemical structure change so as to activate or regulate the structure and the function of the protein. Glycosylation, a major posttranslational modification of proteins, has an important influence on the structure and function of proteins, and the sugar chains of glycoproteins play an important role in cell-to-cell recognition, adhesion, communication, immune response, and the like.

According to the classical natural medicine theory, bioactive substances mainly comprise peptides, saccharides, terpenes, alkaloids, polyethers, macrolides, sterols and the like, and the classification method is only a simple classification. Peptides, saccharides, and not only metabolic products of proteins, but "terpenes, alkaloids, polyethers, macrolides, sterols" are not metabolic products of proteins, but only metabolic products of proteins, and are substances stored in specific cells or cell sites of organisms.

The early major work in the development of natural drugs has been in the fields of terpenes, alkaloids, polyethers, macrolides, sterols, etc. With the development of high performance liquid chromatography, mass spectrometry, and liquid chromatography-mass spectrometry, the development of natural drugs is becoming more focused on peptides and saccharides.

The polypeptide is a compound formed by combining one or more amino acids through peptide bonds according to a certain arrangement sequence, and the structure of the compound is between the amino acids and the protein. Compared with proteins, polypeptides have a simple structure and a small molecular weight, and more readily pass through various biological membranes, and play important roles in life activities, for example, in molecular recognition, signal transduction, cell differentiation, and ontogeny. The polypeptide is better than amino acid in absorption in organism, and has physiological activity function incomparable with amino acid. The source of the polypeptide is divided into two sources of synthesis and natural. The native polypeptide is derived from: animals, plants, microorganisms. The end of part of polypeptide chain is glycosylated and modified to become glycopeptide, however, glycopeptide only occupies a small part (2-5%) of all peptide segments, mass spectrum response is easily inhibited by high abundance non-glycopeptide, and the activities of glycopeptide and polypeptide are not consistent. Polysaccharides are important active substances for maintaining free water required for vital activities in the body, and polypeptides and polysaccharides in the living body play important physiological roles.

The natural polypeptide has antibacterial, antiviral, and blood pressure lowering activities, Mohammad A obtains a cyclic acetal peptide (Microspisamid) composed of 13 amino acid residues from Indonesia sponge, and the compound can inhibit cytopathic effect caused by HIV-I infection. Zhao et al extracted an antihypertensive peptide from tuna skeletal protein hydrolysate (seoul fishery, 2015, vol 32, stage 5). The university of Catharanthus roseus supports the research of the pest-resistant mechanism of the pea plant polypeptide PA1 b.

The research on sugar reduction and pain relief of ginseng glycopeptides is supported by the Chinese medical academy of Jilin province, the research on hepatic fibrosis resistance of turtle shell polypeptide is supported by Hubei Chinese medical university, the molecular mechanism of ascidian polypeptide antagonism to survivin is supported by the oceanic research institute of Chinese academy of sciences, the construction of a novel major disease treatment system based on natural polysaccharide-polypeptide couplet is supported by Beijing chemical university, the molecular mechanism research on rice bran polypeptide mediated cholesterol outflow is supported by Beijing industry and university of Qingdao science and technology supports the molecular mechanism research on the inhibition of lung cancer growth by clam polypeptide targeting tubulin. Southern medical university has supported the structure and function studies of potential medicinal polypeptides from the skin of a hypsizygus marmoreus. University of south China supports the research on American cockroach antiviral polypeptide, and university of Kunming medical science supports the research on the structure and function of potential medicinal polypeptide and protein in the skin of Rana arborina.

The patent application ZL931047226 filed by the university of Hunan university relates to a tiger-stripe bird-catching spider toxin and a preparation method thereof, wherein the chemical structure of the tiger-stripe bird-catching spider toxin is a polypeptide consisting of 33 amino acid residues in a specific sequence and containing three pairs of intra-chain disulfide bonds. It can irreversibly block mammalian neuromuscular junction transmission, and can be used as a tool reagent for neurobiological and pharmacological research.

ZL00042548 is submitted to the university of the Hunan Master, and relates to the application of tiger stripe analgesic peptide in preparing analgesic drugs. The tiger stripe analgesic peptide can be used as a single effective active component for preparing analgesic drugs. The Huwen analgesic peptide blocks the transmission of pain nerve signals by inhibiting the release of presynaptic neurotransmitter through the combination with N-type high threshold calcium ion channels of the presynaptic membrane.

Korean institute of science and technology filed ZL9510017701.2, which relates to a method for purifying hirudin using metal ion affinity chromatography with copper ions as affinity adsorbent and phosphate buffer as eluent.

Zl200380002805.x filed by denna ltd, relates to a process for the preparation of the alpha subunit of β -conglycinin, resuspending the β -conglycinin enriched fraction in denaturing buffer, and performing metal affinity chromatography purification on iminodiacetic acid agarose matrix (Sigma) bound to zinc to obtain the alpha subunit.

ZL200610017237.6 was submitted by Jilin province institute of traditional Chinese medicine and Chinese medicine, and relates to a method for reflux extraction of ginseng with 50% ethanol and 80% ethanol precipitation to obtain ginseng glycopeptide crude product, and simultaneously controlling the approximate range of molecular weight, and then removing impurities by refrigeration, decolorization and ultrafiltration to obtain refined ginseng glycopeptide.

ZL200810230190.0 is submitted to university of Liaoning teachers and relates to a wood frog anticancer peptide and a preparation method thereof, the anticancer peptide takes wood frog skin as a raw material, and is purified by column chromatography, desalted, frozen and condensed after being extracted by acidified ethanol and the supernatant fluid is frozen and dried, so as to obtain the wood frog anticancer peptide with anticancer activity.

ZL200710012966.7 was submitted by the institute of chemical and physical sciences of Chinese academy of sciences, and phosphopeptides are retained in the immobilized titanium ion affinity chromatography material due to the strong interaction between phosphate groups and immobilized titanium ions, thereby realizing the specific separation and enrichment of phosphopeptides from complex proteolytic liquid.

ZL201610442864.8, submitted by the institute of chemical and physical sciences of Chinese academy of sciences, relates to the preparation of a ginseng extract containing endogenous ginseng polypeptide as the main ingredient by grinding ginseng into powder, extracting with an organic solvent or formic acid, separating the extract with a column, and concentrating the eluate under reduced pressure.

The university of Guangxi submitted ZL201810125564.6, which relates to the steps of extracting turtle shells serving as a raw material with water to obtain a reaction raw material solution, performing enzymolysis treatment, microfiltration and ultrafiltration, separating by using a gel chromatography column and an anion exchange chromatography column to obtain an inhibitory peptide raw material, and performing multiple chromatographic separation and purification on the inhibitory peptide raw material to obtain the angiotensin converting enzyme inhibitory peptide.

Although the research on natural polypeptide has been the focus of the global pharmaceutical research, the difficulties encountered in the research and development process are very significant, and especially the purification and isolation techniques are not perfect, so it is necessary to conduct intensive research on the extraction and preparation process of natural polypeptide.

Technical scheme

The invention relates to a medicament for treating respiratory system diseases, which is characterized in that:

treating or preventing pulmonary embolism, or, interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, lung disease caused by smoking, or;

treating or preventing pulmonary embolism, or hemorrhage due to interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, and lung disease caused by smoking.

The invention relates to a medicament for treating respiratory system diseases, which is characterized in that:

the natural raw materials are from: centipede, leech, ground beeltle, periplaneta americana, scorpion, earthworm, Chinese forest frog, ginseng, American ginseng, medlar and lucid ganoderma.

The invention relates to a medicament for treating respiratory system diseases, which is characterized in that:

the preparation method is carried out according to the conventional method.

The invention relates to a medicament for treating respiratory system diseases, which is characterized in that:

the natural raw materials are prepared by any one of drying, crushing, water extraction, ethanol extraction, polyamide separation, macroporous resin separation and sephadex separation and the combination thereof.

The invention relates to a medicament for treating respiratory system diseases, which is characterized in that:

is separated from natural raw materials, and the separation process comprises the following steps:

the first stage is as follows: primary processing of natural raw materials; the method comprises the following steps: drying, crushing and removing impurities;

and a second stage: enriching natural peptides;

and a third stage: purifying the natural peptide: preparing a liquid phase, desalting,

a fourth stage: the preservation comprises the following steps: and (5) freeze-drying.

A medicament for treating respiratory diseases, characterized by: the dosage forms comprise capsule, enteric capsule, tablet, injection, lyophilized powder for injection, controlled release injection, liposome injection, and spray.

A medicament for treating respiratory diseases, characterized by: may also include a pharmaceutical carrier and/or a pharmaceutical excipient.

A medicament for treating respiratory diseases, characterized by: scorpion venom, centipede venom, etc., and can be used for treating or preventing pulmonary embolism, interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, and lung disease caused by smoking; treating or preventing pulmonary embolism, or hemorrhage due to interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, and lung disease caused by smoking.

A medicament for treating respiratory diseases, characterized by: the natural peptide is LPRW peptide, LPRT peptide, LPRR peptide, and LPRL peptide.

Experimental example 1:

the first stage is as follows: preference for primary processing of natural raw materials

The primary processing steps of the natural raw materials of the invention are as follows: drying, pulverizing, and removing impurities.

The drying mode of the natural raw materials is as follows: vacuum drying or natural drying;

the crushing mode of the natural raw materials is as follows: any one of conventional pulverization and superfine pulverization;

the impurities of the natural raw materials are removed as follows: any one of water extraction, ethanol extraction and carbon dioxide supercritical extraction.

Selecting four natural raw materials, removing impurities, spreading, naturally drying for 12 and 24 hours, pulverizing, sieving with 200 mesh sieve, soaking in 85% ethanol for 2 times, each for 12 hours, extracting to remove fat to obtain natural raw material powders A1, A2, A3 and A4;

selecting four natural raw materials, removing impurities, spreading uniformly, vacuum drying for 4, 8 hours, micronizing to 100 μm, microwave puffing, soaking in 85% ethanol for 2 times, each for 12 hours, extracting to remove fat to obtain natural raw material powders B1, B2, B3, and B4;

selecting four natural raw materials, removing impurities, uniformly spreading, vacuum drying for 4-8 hours, micronizing to 100 μm, microwave puffing, and carbon dioxide supercritical extraction to remove fat, wherein the carbon dioxide supercritical extraction process conditions are as follows: the extraction pressure is 25MPa, the extraction temperature is 35 ℃, the flow of CO2 is 20L/h, and the extraction time is 2 h; obtaining natural raw material powders C1, C2, C3 and C4;

the natural raw materials are as follows: centipede, scorpion, ginseng, ganoderma lucidum each 20 g.

And (4) measuring the content of impurities in the groups A, B and C of the natural raw material powder.

The method adopts group C process for Scolopendra, Scorpio, Ganoderma, and Ginseng radix, and has high impurity yield.

Experimental example 2: enrichment of natural peptides

And a second stage: enriching natural peptides: eluting according to 1, or, 2, or, 3 times

Step 1, taking natural raw material powder (C1, C2, C3 and C4), adding a proper amount of chitosan, dissolving with distilled water, adsorbing suspension by adopting polyamide, eluting with distilled water and 5-15% ethanol, collecting ethanol eluate, and adjusting the ethanol content of the eluate to 2-4%; (ii) a

Step 2, adsorbing 2-4% ethanol eluent by using macroporous resin, eluting by using distilled water and 5-15% ethanol, and collecting ethanol eluent;

any of 1 and 2 may be repeated.

The polyamide and the macroporous absorption resin are 80-200 meshes.

The macroporous adsorbent resin is medium-polar, weak-polar or nonpolar adsorbent resin, such as D101, HPD100, AB-8, HPD400, HPD600, etc.,

collecting the eluates of the first and second stages 1-step distilled water from the natural raw material powders (C1, C2, C3, and C4), and vacuum drying.

Collecting the ethanol eluate of the step 1 of the second stage in the obtained natural raw material powder (C1, C2, C3, C4), and vacuum drying to obtain vacuum dried powder.

Experimental example 3: purifying the natural peptide: preparation of liquid phase, desalting

Preparation chromatography: bestarose HP, DEAE Bestarose FF, Sephadex TMG-50, Sephadex TMG-75, Sephadex LH-20, CM Sepharose TM, Bestarose FF, 200mm × 250 mm; 200mm × 600 mm; 200mm × 400 mm; 200mm x 800mm

Sample injection: vacuum drying deionized water solution of the powder;

mobile phase: 0.01mol/L sodium hydroxide aqueous solution (A),

0.01mol/L potassium dihydrogen phosphate water solution (B),

gradient elution procedure:

0-10min，B:5％

10-20min，B:5-20％；

collecting the peak time; part of 10-19 min;

flow rate: 5.0-10.0mL/min,

detection wavelength: the particle size of the nano-particles is 260nm,

sample introduction amount: 0.5mL of the solution is added,

collecting the dehydrated and desalted extract.

Experimental example 4: determination of the lyophilization Process

Determination of minimum melting Point

The instrument comprises the following steps: SWCII digital Beckmann thermometer and freezing point tester

The method comprises the following steps: placing the dehydrated and desalted extract in an inner tube of a freezing point tester, inserting a temperature thermocouple, and freezing in a refrigerator at-25 deg.C. And (3) placing the frozen sample in an outer tube of a freezing point tester, measuring the temperature of the sample at different times, and drawing a sample temperature-time curve. The lowest melting point is read from the curve.

The freeze-drying process is carried out according to the following degrees: firstly, a pre-freezing process; ② sublimation drying; and thirdly, resolving and drying.

The freeze drying process is optimized by taking the appearance formability, redissolution property and water content as the investigation indexes.

Compared with the prior art, the invention has the following advantages: the natural peptide provided by the invention has the advantages of wide source, high purity, good stability and strong activity;

the sources of starfish, cow frog skin, centipede, leech, ground beeltle, American cockroach, scorpion, earthworm, Chinese forest frog, ginseng, American ginseng, medlar and lucid ganoderma comprise commercial sources or are prepared according to the common technology.

Example 1: preparation of LPRW peptide and its freeze dried powder

Preparation of centipede raw material powder: selecting 500 g centipedes, removing impurities, uniformly spreading, drying for 4 hours in vacuum, carrying out superfine grinding for 100 mu m, carrying out microwave puffing, carrying out carbon dioxide supercritical extraction for removing fat, wherein the process conditions of the carbon dioxide supercritical extraction are as follows: the extraction pressure is 25MPa, the extraction temperature is 35 ℃, the flow of CO2 is 20L/h, and the extraction time is 2 h; obtaining one of centipede raw material powder;

dissolving 100g of centipede raw material powder in distilled water, adsorbing by macroporous resin D101 with 200 meshes, eluting by distilled water and 10% ethanol, and collecting ethanol eluate; vacuum drying to obtain one of Scolopendra vacuum dried powder.

Taking 3 g of one of centipede vacuum drying powder, adding 0.1 g of chitosan, dissolving with deionized water, and refining by preparative column chromatography, wherein a filler is Sephadex LH-20; sample injection: vacuum drying deionized water solution of the powder; mobile phase: 0.01mol/L aqueous solution of sodium hydroxide (A) and 0.01mol/L aqueous solution of potassium dihydrogen phosphate (B); gradient elution procedure: 0-10min, B5%; 10-20min, B10%; collecting the peak time; part of 10-12 min; flow rate: 1.0mL/min, detection wavelength: 260nm, sample size: 0.5 mL; dehydrating and desalting to obtain one of Scolopendra refined products;

adding 5.0mg methionine and 5.0g lactose into 10.0mg of one of Scolopendra refined extract, dissolving, adding water for injection, heating to dissolve, diluting to 100ml, filtering, ultrafiltering the filtrate, freezing at-10 deg.C for 5 hr, drying under reduced pressure for 3 hr and-20 deg.C; heating and drying for 4 hours at the temperature of minus 5 ℃, and heating and drying for 4 hours for the second time at the temperature of 20 ℃; one of the white powder LPRW peptide freeze-dried powder is obtained.

Example 2: preparation of LPRW peptide II and its freeze dried powder

Preparation of centipede raw material powder: selecting 500 g centipedes, removing impurities, uniformly spreading, drying for 4 hours in vacuum, carrying out superfine grinding for 100 mu m, carrying out microwave puffing, carrying out carbon dioxide supercritical extraction for removing fat, wherein the process conditions of the carbon dioxide supercritical extraction are as follows: the extraction pressure is 25MPa, the extraction temperature is 35 ℃, the flow of CO2 is 20L/h, and the extraction time is 2 h; obtaining the second centipede raw material powder;

taking two 100g of centipede raw material powder, adding 5g of chitosan, dissolving with distilled water, adsorbing suspension by using 200-mesh polyamide, eluting with distilled water and 10% ethanol, collecting ethanol eluate, drying in vacuum, dissolving with distilled water, adsorbing with 200-mesh macroporous resin D101, eluting with distilled water and 10% ethanol, and collecting ethanol eluate; vacuum drying to obtain Scolopendra powder II.

Taking 3 g of centipede vacuum drying powder, adding 0.1 g of chitosan, dissolving with deionized water, and refining by preparative column chromatography, wherein a filler is Sephadex LH-20; sample injection: vacuum drying deionized water solution of the powder; mobile phase: 0.01mol/L aqueous solution of sodium hydroxide (A) and 0.01mol/L aqueous solution of potassium dihydrogen phosphate (B); gradient elution procedure: 0-10min, B5%; 10-20min, B10%; collecting the peak time; part of 10-12 min; flow rate: 1.0mL/min, detection wavelength: 260nm, sample size: 0.5 mL; dehydrating and desalting to obtain the second centipede refined product;

adding 5.0mg methionine and 5.0g lactose into the second 10.0mg of Scolopendra refined extract, dissolving, adding water for injection, heating to dissolve, diluting to 100ml, filtering, ultrafiltering the filtrate, freezing at-10 deg.C for 5 hr, drying under reduced pressure for 3 hr and-20 deg.C; heating and drying for 4 hours at the temperature of minus 5 ℃, and heating and drying for 4 hours for the second time at the temperature of 20 ℃; the second lyophilized powder of white LPRW peptide was obtained.

Example 3: preparation of one of LPRT peptides and freeze-dried powder thereof

Preparing scorpion raw material powder: selecting 500 g of scorpion, removing impurities, uniformly spreading, drying for 4 hours in vacuum, carrying out superfine grinding for 100 mu m, carrying out microwave puffing, carrying out carbon dioxide supercritical extraction for removing fat, wherein the process conditions of the carbon dioxide supercritical extraction are as follows: the extraction pressure is 25MPa, the extraction temperature is 35 ℃, the flow of CO2 is 20L/h, and the extraction time is 2 h; obtaining one of the scorpion raw material powder;

dissolving one 100g of Scorpio powder in distilled water, adsorbing with 200 mesh macroporous resin D101, eluting with distilled water and 10% ethanol, and collecting ethanol eluate; vacuum drying to obtain one of Scorpio vacuum dried powder.

Taking 2g of one of the scorpion vacuum drying powder, adding 0.1 g of chitosan, dissolving by deionized water, and refining by preparative column chromatography, wherein the filler is Sephadex LH-20; sample injection: vacuum drying deionized water solution of the powder; mobile phase: 0.01mol/L aqueous solution of sodium hydroxide (A) and 0.01mol/L aqueous solution of potassium dihydrogen phosphate (B); gradient elution procedure: 0-10min, B5%; 10-20min, B10%; collecting the peak time; 15-18 min; flow rate: 1.0mL/min, detection wavelength: 260nm, sample size: 0.5 mL; dehydrating and desalting to obtain one of Scorpio refined products;

adding methionine 5.0mg into one of Scorpio refined extract 10.0mg, adding lactose 5.0g for dissolving, adding water for injection, heating for dissolving, diluting to 100ml, filtering, ultrafiltering the filtrate, freezing at-10 deg.C for 5 hr, drying under reduced pressure for 3 hr and-20 deg.C; heating and drying for 4 hours at the temperature of minus 5 ℃, and heating and drying for 4 hours for the second time at the temperature of 20 ℃; one of the white powder of the lyophilized powder of LPRT peptide was obtained.

Example 4: preparation of LPRT peptide II and its freeze dried powder

Preparing a second scorpion raw material powder: selecting 500 g of scorpion, removing impurities, uniformly spreading, drying for 4 hours in vacuum, carrying out superfine grinding for 100 mu m, carrying out microwave puffing, carrying out carbon dioxide supercritical extraction for removing fat, wherein the process conditions of the carbon dioxide supercritical extraction are as follows: the extraction pressure is 25MPa, the extraction temperature is 35 ℃, the flow of CO2 is 20L/h, and the extraction time is 2 h; obtaining the second scorpion raw material powder;

taking two 100g of scorpion raw material powder, adding 5g of chitosan, dissolving with distilled water, adsorbing suspension by using 200-mesh polyamide, eluting with distilled water and 10% ethanol, collecting ethanol eluate, vacuum drying, dissolving with distilled water, adsorbing with 200-mesh macroporous resin D101, eluting with distilled water and 10% ethanol, and collecting ethanol eluate; vacuum drying to obtain Scorpio vacuum dried powder II.

Taking two 2g of the scorpion vacuum drying powder, adding 0.1 g of chitosan, dissolving by deionized water, and refining by preparative column chromatography, wherein the filler is Sephadex LH-20; sample injection: vacuum drying deionized water solution of the powder; mobile phase: 0.01mol/L aqueous solution of sodium hydroxide (A) and 0.01mol/L aqueous solution of potassium dihydrogen phosphate (B); gradient elution procedure: 0-10min, B5%; 10-20min, B10%; collecting the peak time; 15-18 min; flow rate: 1.0mL/min, detection wavelength: 260nm, sample size: 0.5 mL; dehydrating and desalting to obtain second Scorpio refined product;

adding methionine 5.0mg into the second 10.0mg of Scorpio refined extract, dissolving lactose 5.0g, adding water for injection, heating to dissolve, diluting to 100ml, filtering, ultrafiltering the filtrate, freezing at-10 deg.C for 5 hr, drying under reduced pressure for 3 hr and-20 deg.C; heating and drying for 4 hours at the temperature of minus 5 ℃, and heating and drying for 4 hours for the second time at the temperature of 20 ℃; obtain white powder LPRT peptide freeze-dried powder II.

Example 5: stability test

The products of examples 1, 2, 3 and 4 were subjected to influence factor tests, accelerated tests and long-term tests according to the guidelines of the XIX C drug stability test in the second part of the "chinese pharmacopoeia", 2010 edition. The investigation items include: properties, identification and content.

The products of examples 1, 2, 3 and 4 were irradiated under strong light for 192 hours, and the content was not changed after the test.

The products of examples 1, 2, 3 and 4 were placed in a retention chamber at 35 ℃ and 60. + -. 10% relative humidity and sampled for determination at 6, 12, 24 and 48 months, respectively, and the contents were unchanged after the test.

Example 6: toxicity test

The products of examples 1, 2, 3, 4 were administered intravenously to rats, dissolved and diluted with 0.9% sodium chloride injection to the concentrations used in the tests, and the products of examples 1, 2, 3, 4 were administered intramuscularly to rats, dissolved and diluted with 0.9% sodium chloride injection to the concentrations used in the tests. The products of examples 1, 2, 3, 4 are non-toxic.

Example 7:

preparation of the product tablet of example 1

0.5g of the product of example 1; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; 2g of magnesium stearate, granulating, drying at 60 ℃, grading by using a 12-mesh sieve, and pressing into tablets.

Preparation of the product capsules of example 1

0.5g of the product of example 1; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; granulating with 100 mesh sieve, drying at 80 deg.C, grading with 100 mesh sieve, and filling into empty capsule.

Preparation of product spray of example 1

0.5g of the product of example 1; adding 30% ethanol water solution, and making into spray according to conventional method.

Preparation of product tablets of example 2

0.5g of the product of example 2; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; 2g of magnesium stearate, granulating, drying at 60 ℃, grading by using a 12-mesh sieve, and pressing into tablets.

Preparation of the product capsules of example 2

0.5g of the product of example 2; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; granulating with 100 mesh sieve, drying at 80 deg.C, grading with 100 mesh sieve, and filling into empty capsule.

Preparation of product spray of example 2

0.5g of the product of example 2; adding 30% ethanol water solution, and making into spray according to conventional method.

Example 8:

preparation of product tablets of example 3

0.5g of the product of example 3; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; 2g of magnesium stearate, granulating, drying at 60 ℃, grading by using a 12-mesh sieve, and pressing into tablets.

Preparation of the product capsules of example 3

0.5g of the product of example 3; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; granulating with 100 mesh sieve, drying at 80 deg.C, grading with 100 mesh sieve, and filling into empty capsule.

Preparation of product spray of example 3

0.5g of the product of example 3; adding 30% ethanol water solution, and making into spray according to conventional method.

Preparation of the product tablet of example 4

0.5g of the product of example 4; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; 2g of magnesium stearate, granulating, drying at 60 ℃, grading by using a 12-mesh sieve, and pressing into tablets.

Preparation of the product capsules of example 4

0.5g of the product of example 4; 10g of microcrystalline cellulose; 10g of poloxamer; 50ml of 95% ethanol; 50g of hydroxypropyl cellulose; 100ml of 25 percent starch slurry; granulating with 100 mesh sieve, drying at 80 deg.C, grading with 100 mesh sieve, and filling into empty capsule.

Preparation of product spray of example 4

0.5g of the product of example 4; adding 30% ethanol water solution, and making into spray according to conventional method.

Example 9

Selecting 500 g centipedes, removing impurities, uniformly spreading, drying in vacuum for 4 hours, and then preparing according to any one of the following modes: crushing; micronizing; extracting with water; or, extracting with ethanol; obtaining centipede extract; the application of the centipede extract in preparing medicaments for treating or preventing respiratory diseases.

Example 10

Selecting 500 g centipedes, removing impurities, uniformly spreading, drying for 4 hours in vacuum, crushing, extracting with water, and drying the water extract in vacuum; passing through macroporous resin column, eluting with purified water, and eluting with 5%, 10%, 20%, 40%, and 80% ethanol; collecting 5% and 10% ethanol-eluted Scolopendra extract; the application of the centipede extract in preparing medicaments for treating or preventing respiratory diseases.

2g of centipede poison, sterilizing and freeze-drying; the application of the compound in preparing medicines for treating or preventing respiratory diseases.

Example 11

Selecting 500 g of scorpion, removing impurities, uniformly spreading, drying in vacuum for 4 hours, and then preparing according to any one of the following modes: crushing; micronizing; extracting with water; or, extracting with ethanol; obtaining the scorpion extract; the Scorpio extract can be used for preparing medicine for treating or preventing respiratory system diseases.

2g of scorpion venom, sterilizing and freeze-drying; the application of the compound in preparing medicines for treating or preventing respiratory diseases.

Example 12

Selecting 500 g of scorpion, removing impurities, evenly spreading, drying for 4 hours in vacuum, crushing, extracting with water, and drying the water extract in vacuum; passing through macroporous resin column, eluting with purified water, and eluting with 5%, 10%, 20%, 40%, and 80% ethanol; collecting 5%, 10%, 20%, 40%, 80% ethanol-eluted Scorpio extract; the Scorpio extract can be used for preparing medicine for treating or preventing respiratory system diseases.

Example 13

Selecting 50g of scorpion and 50g of centipede, removing impurities, uniformly spreading, drying in vacuum for 4 hours, and then preparing according to any one of the following modes: crushing; micronizing; extracting with water; or, extracting with ethanol; obtaining scorpion centipede extract; the Scorpio and Scolopendra extract can be used for preparing medicine for treating or preventing respiratory system diseases.

Example 14

Selecting 50g of scorpion and 50g of centipede, removing impurities, uniformly spreading, drying in vacuum for 4 hours, crushing, extracting with water, and drying the water extract in vacuum; passing through macroporous resin column, eluting with purified water, and eluting with 5%, 10%, 20%, 40%, and 80% ethanol; collecting 5% and 10% ethanol-eluted Scorpio-Scolopendra extract; the Scorpio and Scolopendra extract can be used for preparing medicine for treating or preventing respiratory system diseases.

Example 15

60 male Wistar rats are taken, 7.5 percent chloral hydrate is injected into the abdominal cavity of 0.6mL/100g for anesthesia, and 2.5U/kg of bleomycin is injected into the trachea. The specific scheme is as follows: weighing and recording the weight of the rat, fixing the rat on an operation table, disinfecting the neck with 70% alcohol, vertically scratching the neck of the rat by a scalpel for about 1cm, separating tissues by using a micro forceps to expose the trachea, penetrating a syringe into the trachea from the annular gap of the tracheal cartilage towards the centripetal end, slowly injecting a bleomycin physiological saline solution with the volume suitable for the weight of the rat according to the measurement of 2.5U/kg, immediately standing the animal, rotating the animal left and right, and uniformly distributing the liquid medicine in the lung.

Referring to the literature method, after adapting to the environment in a clean-grade observation room for 3 days, 36 mice are selected and randomly divided into 6 groups, wherein each group comprises 6 mice, the first group is a blank control group, the second group is a group of 80mg/kg injection for removing thromboembolism, the third group is a group of 20ug/kg lyophilized powder of LPRW peptide, the fourth group is a group of 20ug/kg lyophilized powder of LPRT peptide, the fifth group is a group of 20ug/kg lyophilized powder of LPRT peptide, the sixth group is a group of 200ug/kg scorpion venom BMK, and the 1-time intraperitoneal injection is carried out, and the volume is 10 ml/kg.

30min after administration, 3% chloral hydrate 300mg/kg was anesthetized by intraperitoneal injection, and the common carotid artery was isolated. And (3) measuring the thrombosis time in the rat body by adopting a YLS-14B type small animal thrombogenesis instrument. The experimental conditions are as follows: the stimulation current is 1.2mA, the stimulation time is 5min, and the recording time is 30 min. When in measurement, the stimulation and temperature control electrodes are fully contacted with the common carotid artery, after the signal is stable, the stimulation is started, the 100 percent formation time of the thrombus in 30min is taken as the formation time of the thrombus in the rat body, and the formation time is taken as '30' if the thrombus does not reach 100 percent formation in 30 min; the maximum percentage of thrombus formation within 30min and 100% thrombus formation within 30min were also recorded.

And (5) carrying out HE (high intensity staining) on the lung tissue section, and observing the pathological characteristics of the lung tissue and the hyperplasia condition of the pulmonary artery intima.

Effect of thrombogenic time in rats

Note: p < 0.05 compared to control blank; p < 0.01; p < 0.001

The rats are administrated by one-time intraperitoneal injection, and compared with a blank control group, the thrombus formation time in the rats can be obviously prolonged by each sample administration group; in addition, the lyophilized powder of LPRW peptide, LPRT peptide and BMK can also obviously reduce the maximum percentage of thrombus formation in rats within 30min and 100% of thrombus formation rate within 30min, and have significant differences compared with a blank control group. The three measurement indexes are comprehensively compared, wherein the effect of the lyophilized powder of LPRT peptide is optimal.

Severe congestion and bleeding of the lungs of the blank control group occurred; inflammatory cells are in a perivascular infiltration around the pulmonary vessels. The alveolar walls of the blank control group are thickened, the alveolar space of the blank control group is obviously increased, and emphysema and edema occur; capillary congestion of alveolar walls of the blank control group, and degeneration, necrosis and shedding of a large number of alveolar epithelial cells and macrophages in alveolar cavities of the blank control group.

Compared with the blank group, the Xuesaitong injection, the freeze-dried powder of LPRW peptide, the freeze-dried powder of LPRT peptide and the scorpion venom BMK can reduce the congestion and bleeding of the lung, reduce the alveolar septum, and reduce the degeneration, necrosis and shedding of alveolar epithelial cells and macrophages, and the freeze-dried powder of LPRT peptide has more obvious effect.

The literature: influence of the zhongjun, zhanghong, etc. coronary artery dredging capsule on experimental thrombosis in rats, journal of chinese experimental prescriptions, 2012; 18(7): 229-230; jurigo, etc., blood-clearing and blood-circulation-promoting granules improve microcirculation and influence on thrombosis, and experimental animal science, 2017; 24(1): 41-44.

Claims (9)

1. A medicament for treating respiratory diseases, characterized by:

treating or preventing pulmonary embolism, or, interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, lung disease caused by smoking, or;

treating or preventing pulmonary embolism, or hemorrhage due to interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, and lung disease caused by smoking.

2. A medicament for the treatment of respiratory diseases according to claim 1, characterized in that:

the natural raw materials are from: centipede, leech, ground beeltle, periplaneta americana, scorpion, earthworm, Chinese forest frog, ginseng, American ginseng, medlar and lucid ganoderma.

3. A medicament for the treatment of respiratory diseases according to claim 2, characterized in that:

the preparation method is carried out according to the conventional method.

4. A medicament for the treatment of respiratory diseases according to claim 2, characterized in that:

the natural raw materials are prepared by any one of water extraction, ethanol extraction, macroporous resin separation and sephadex separation and the combination thereof.

5. A medicament for the treatment of respiratory diseases according to claim 4, characterized in that:

is separated from natural raw materials, and the separation process comprises the following steps:

the first stage is as follows: primary processing of natural raw materials; the method comprises the following steps: drying, crushing and removing impurities;

and a second stage: enriching natural peptides;

and a third stage: purifying the natural peptide: preparing a liquid phase, desalting,

a fourth stage: the preservation comprises the following steps: and (5) freeze-drying.

6. A medicament for the treatment of respiratory diseases according to claim 1, characterized in that: the dosage forms comprise capsule, enteric capsule, tablet, injection, lyophilized powder for injection, controlled release injection, liposome injection, and spray.

7. A medicament for the treatment of respiratory diseases according to claims 1-3, characterized in that: may also include a pharmaceutical carrier and/or a pharmaceutical excipient.

8. A medicament for the treatment of respiratory diseases according to claims 1-3, characterized in that: the natural peptide is LPRW peptide, LPRT peptide, LPRR peptide, and LPRL peptide.

9. A medicament for treating respiratory diseases, characterized by: scorpion venom and centipede venom for preparing medicine for treating or preventing pulmonary embolism, interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis and lung disease caused by smoking; can be used for preparing medicine for treating or preventing pulmonary embolism, or, interstitial pneumonia, viral pneumonia, pulmonary edema, emphysema, silicosis, pneumoconiosis, and hemorrhage complicated with lung disease caused by smoking.