CN108434166B

CN108434166B - Pharmaceutical composition for treating thromboembolism, preparation method, preparation and application thereof

Info

Publication number: CN108434166B
Application number: CN201810227117.1A
Authority: CN
Inventors: 周荣光; 赵加强; 顾静波; 文冰亭; 郭泽剑
Original assignee: Kunming Pharmaceutical Corp
Current assignee: Kunming Pharmaceutical Corp
Priority date: 2018-03-20
Filing date: 2018-03-20
Publication date: 2021-06-22
Anticipated expiration: 2038-03-20
Also published as: CN108434166A

Abstract

The invention discloses a pharmaceutical composition for treating thromboembolism, a preparation method, a preparation and application thereof. The pharmaceutical composition for treating the thromboembolism comprises the following pharmaceutical components in percentage by mass: ginsenoside Rb20.001-99.996%, ginsenoside Rb10.001-99.996%, ginsenoside Rg10.001-99.996%, ginsenoside Rc 0.001-99.996% and notoginsenoside R10.001-99.996%. The pharmaceutical composition and the preparation thereof have obvious curative effects on ischemic cardiovascular and cerebrovascular diseases such as myocardial infarction, cerebral apoplexy, cerebral infarction, cerebral hemiplegia and the like, and retinopathy or optic nerve injury diseases caused by the ischemic cardiovascular and cerebrovascular diseases, diabetes and the like, and have wide application prospects. Compared with the prior art and products, the pharmaceutical composition and the preparation thereof for treating the thromboembolism have better curative effect and higher safety, and can better meet the requirements of clinical medication. The preparation method provided by the invention has the advantages of simple process, mild conditions, convenience in operation, stable and controllable quality, and suitability for industrial large-scale production.

Description

Pharmaceutical composition for treating thromboembolism, preparation method, preparation and application thereof

Technical Field

The invention belongs to the technical field of medicines, and particularly relates to a Xuesaitong medicinal composition, and a preparation method, a preparation and application thereof.

Background

Pseudo-ginseng is the dry root and rhizome of the Araliaceae plant Panax Notoginseng (Burk) F. H Chen, also named pseudo-ginseng and pseudo-ginseng, is a rare traditional Chinese medicine for treating cardiovascular and cerebrovascular diseases in traditional Chinese medicine, and has the effects of removing blood stasis, stopping bleeding, reducing swelling and relieving pain. Recorded in Yu Jia Yao Jie, san Qi can harmonize Ying and stop bleeding, unblock collaterals and remove blood stasis, and remove blood stasis to astringe new blood. All blood stasis is broken after delivery, in menstrual period, traumatic injuries, carbuncle and swelling; it can stop any new blood, such as hematemesis, epistaxis, metrorrhagia, knife wound, and flechage wound. "

Panax Notoginsenosides (PNS) are a general term for a series of saponins and aglycones thereof extracted from Panax notoginseng, and the constituents are diverse and complex, some constituents have been separated and confirmed, but a large number of unknown constituents have not been clarified by human beings. To date, over 100 monomeric saponin components have been isolated and identified from PNS, such as ginsenosides Rb1, Rb2, Rb3, Rc, Rd, Re, Ro, Rh1, Rg1, Rg2, Rg3, Rg4, Rg5, notoginsenoside R1, R2, R3, R4, and the like. These saponins are mainly classified into triol type and diol type according to their aglycones. The diol type (PDS) mainly comprises panaxadiol saponins Rb1, Rb2, Rc, Rd, Rh2, etc., and the triol type (PTS) mainly comprises panaxatriol saponins Re, Rg1, Rg2, Rh1, R1, etc. The content of notoginsenoside in the cultivated roots is reported to be Rg1(43.36%), Rb1(41.08%), R1(3.65%), Re (3.42%), Rh1 (0.22%) and other saponin 8.26% from high to low. Literature studies have also shown that PNS contains mainly ginsenoside Rb1 (about 30%), ginsenoside Rg1 (about 20%), notoginsenoside R1 (about 5%) and ginsenoside Re (about 2.5%). It is also known that the underground part of notoginseng is mainly triol saponin, and the content ratio of triol saponin to diol type is about 3: 1, while the overground part is mainly diol type (PDS).

The Xuesaitong product sold in the market at present is a traditional Chinese medicine preparation which is prepared by taking Panax Notoginseng Saponins (PNS) as a medicine raw material and adding necessary pharmaceutic adjuvants. The Xuesaitong products which are on the market at present have the dosage forms of freeze-drying agent, injection, capsule, soft capsule, tablet, dropping pill and the like, have the efficacies of activating blood and dissolving stasis, and dredging and activating collaterals, and are clinically used for treating the meridian restoration stage of ischemic apoplexy (cerebral infarction) caused by blood stasis blockage, and the symptoms of hemiplegia, hemianesthesia, facial distortion, aphasia and the like. However, in the current Panax Notoginseng Saponins (PNS) raw materials from different manufacturers on the market, due to different production places, different application positions, different harvesting time and years of panax notoginseng used for extracting the panax notoginseng saponins, different extraction methods and different process conditions, the obtained Panax Notoginseng Saponins (PNS) have different chemical components, and even if different batches of Panax Notoginseng Saponins (PNS) are produced by the same manufacturer, the chemical components and the composition and the content of the panax notoginseng saponins are different. Due to the reasons, the pharmacodynamic components and the compositions of the commercially available Xuesaitong products are not fixed and clear, and the pharmacodynamic components and the composition ratios of the products of different manufacturers or different batches are different, so that the clinical curative effect of the products is seriously influenced, and a series of adverse reactions and safety problems are caused, and therefore, research and solution by scientific researchers are urgently needed.

Disclosure of Invention

The first purpose of the invention is to provide a pharmaceutical composition for treating thrombus; the second purpose is to provide a preparation method of the pharmaceutical composition for treating thromboembolism; the third purpose is to provide the preparation of the pharmaceutical composition for treating the thromboembolism; the fourth purpose is to provide a soft capsule preparation of the pharmaceutical composition for treating the thromboembolism; the fifth purpose is to provide a preparation method of the soft capsule preparation of the pharmaceutical composition for treating thromboembolism; the sixth purpose is to provide the application of the pharmaceutical composition for treating thromboembolism and/or the preparation thereof.

The first purpose of the invention is realized by that the pharmaceutical composition for treating thromboembolism comprises the following pharmaceutical components in percentage by mass:

ginsenoside Rb20.001-99.996%

Ginsenoside Rb10.001-99.996%

Ginsenoside Rg 10.001-99.996%

Ginsenoside Rc 0.001-99.996%

And (3) notoginsenoside R10.001-99.996%.

Preferably, the pharmaceutical composition comprises the following monomeric pharmaceutical components in percentage by mass:

ginsenoside Rb20.01-99.96%

Ginsenoside Rb10.01-99.96%

Ginsenoside Rg 10.01-99.96%

Ginsenoside Rc 0.01-99.96%

10.01 to 99.96 percent of notoginsenoside R

More preferably, the pharmaceutical composition comprises the following monomer pharmaceutical components in percentage by mass:

ginsenoside Rb254.6%

Ginsenoside Rb121.1%

Ginsenoside Rg 111.3%

Ginsenoside Rc 8.5%

Notoginsenoside R14.5%

Or:

ginsenoside Rb220.5%

Ginsenoside Rb147.8%

Ginsenoside Rg123.4%

Ginsenoside Rc 6.7%

Notoginsenoside R11.6%

Or:

ginsenoside Rb21.2%

Ginsenoside Rb114.6%

Ginsenoside Rg 155.7%

Ginsenoside Rc 19.3%

Notoginsenoside R19.2%

Or:

ginsenoside Rb221.1%

Ginsenoside Rb12.5%

Ginsenoside Rg 128.2%

Ginsenoside Rc 37.6%

Notoginsenoside R110.6%

Or:

ginsenoside Rb220.9%

Ginsenoside Rb12.7%

Ginsenoside Rg 121.5%

Ginsenoside Rc 10.2%

Notoginsenoside R144.7%

In the invention, the ginsenoside Rb2 is a monomer compound, and the chemical structure is shown as the following formula

Shown as the following, the English name is Ginsenoside Rb2, the English chemical name is 20- ((6-O-alpha-L-Arabinopyranosyl-beta-D-glucopyranosyl) oxy) -12beta-hydroxydammar-24-en-3beta-yl 2-O-beta-D-glucopyranosyl-beta-D-glucopyranoside, the molecular formula is C₅₃H₉₀O₂₂Molecular weight 1079.27;

the chemical structure of ginsenoside Rb1 is shown in the following formula

Shown as the English name Ginsenoside Rb1, the English chemical name is (3beta,12beta) -20- [ (6-O-beta-D-Glucopyranosyl) oxy]-12-hydroxydammar-24-en-3-yl2-O-beta-D-glucopyranosyl-beta-D-glucopyranoside, molecular formula C54H92O23, molecular weight 1109.29;

the chemical structure of the ginsenoside Rg1 is shown as the following formula

Shown in the formula of the Chinese, the English name is Ginsenoside Rg1, the alias is notoginsenoside C1, the English alias is Sanchinoside C1, and the molecular formula is C₄₂H₇₂O₁₄Molecular weight 801.01;

ginsenoside Rc has the chemical structure shown in the following formula

Shown as the English name of Ginsenoside Rc, the English chemical name of 20- [ (6-O-alpha-L-Arabinofuranosyl-beta-D-glucopyranosyl) oxy]-12b-hydroxydammar-24-en-3b-yl 2-O-beta-D-glucopyranosyl-beta-D-glucopyranoside, molecular formula C₅₃H₉₀O22, molecular weight 1079.27;

notoginsenoside R1, chemical structure shown in the following formula

Shown in the specification, the chemical name of the compound is Notogenoside R1, the chemical name of the compound is Glucopyranosyloxy) -3, 12-dihydroxdalmar-24-en-6-yl 2-O-beta-D-glucopyranosyl-, (3beta, 6 alpha, 12beta) -20- (beta-D-Glucopyranosyloxy) -3, 12-dihydroxdalmar-24-en-6-yl 2-O-beta-D-glucopyranosyl-beta-D-glucopyranoside, the molecular formula of which is C₄₇H₈₀O₁₈Molecular weight 933.131.

The compounds of ginsenoside Rb2, ginsenoside Rb1, ginsenoside Rg1, ginsenoside Rc and notoginsenoside R1 can be extracted and separated from plants such as Notoginseng radix, Ginseng radix, radix Panacis Quinquefolii, radix Pseudostellariae, radix Acanthopanacis Senticosi, rhizoma Gynurae Divaricatae, rhizoma Panacis japonici, and Korean ginseng, or prepared by total synthesis, semi-synthesis or biosynthesis. At present, the product can be directly purchased from the market, and the purity can reach more than 98 percent.

The invention takes the commercial panax notoginseng saponins medicine as the contrast, adopts ICR mice to carry out intraperitoneal injection administration, carries out the acute toxicity experimental study of intravenous injection administration of the medicine composition, and the result shows that: the LD50 of the pharmaceutical composition ICR mouse by single intravenous injection is 397mg/kg, which is obviously higher than the LD50 value of the commercial panax notoginseng saponins, namely 306mg/kg, which shows that the pharmaceutical composition has extremely high medicinal safety and the safety is superior to that of the existing panax notoginseng saponins. See example 4 for details.

In addition, the invention also takes the commercial panax notoginseng saponins as the contrast, and carries out the experimental study on the vascular irritation, the hemolytic property and the anaphylaxis of the pharmaceutical composition, and the result shows that the pharmaceutical composition has no obvious adverse reactions such as irritation, hemolytic property, anaphylaxis and the like, completely meets the requirement of medicinal safety, and each adverse reaction safety evaluation index is superior to the panax notoginseng saponins contrast drug. See example 5 for details.

The second object of the present invention is achieved by comprising the steps of:

step 1), weighing notoginsenoside R1, placing the notoginsenoside R1 in an ethanol solvent, and stirring and dissolving to obtain a solution A;

step 2) weighing ginsenoside Rb2, ginsenoside Rb1, ginsenoside Rg1 and ginsenoside Rc, dissolving in distilled water under stirring, heating to 50-70 deg.C, adding solution A under stirring, adding active carbon for injection, heating to boil for 30-50 min, standing, cooling to room temperature, and filtering to remove active carbon;

and 3) placing the filtrate in a vacuum freeze dryer, cooling to-40 to-30 ℃, pre-freezing for 3-5 hours, controlling the temperature to-25 to-20 ℃ and the pressure to 10-20 Pa, carrying out sublimation drying for 3-5 hours, controlling the temperature to 30-50 ℃ and the pressure to 1-10 Pa, and carrying out resolution drying for 4-6 hours to obtain the product.

The traditional preparation method of the pharmaceutical composition is usually realized by adopting ways of mechanical grinding, mixing and the like, and has the problems that various components are difficult to uniformly mix, the quality of the product is lack of uniformity and the like. Especially, the raw materials are subjected to the action of mechanical shearing force and impact force in the mixing and grinding processes, and the raw materials rub against each other to generate local overheating, so that the stability and the pharmacological activity of the raw materials are damaged to a certain extent. In order to overcome the defects, the pharmaceutical composition is prepared by adopting a solvent dissolving and low-temperature freeze drying mode, so that the quality uniformity of the composition is ensured, and the stability and the pharmacological activity of the pharmaceutical ingredients are fully ensured without being damaged.

The third purpose of the invention is realized by that the pharmaceutical preparation for treating thromboembolism is prepared from the pharmaceutical composition for treating thromboembolism and pharmaceutical auxiliary materials, carriers and matrixes, and comprises injections, tablets, oral liquid, capsules, soft capsules, dropping pills, sustained-release preparations and controlled-release preparations.

The pharmaceutical preparation for treating the thromboembolism is prepared from the pharmaceutical composition for treating the thromboembolism and pharmaceutical auxiliary materials, carriers and matrixes, the administration route can be intestinal or parenteral, such as oral administration, intravenous injection, intramuscular injection, subcutaneous injection, nasal cavities, oral mucosa, eyes, lungs, respiratory tracts, skin, vagina, rectum and the like, and the dosage form can be tablets, capsules, soft capsules, granules, pills, dropping pills, injection, freeze-dried powder injection, oral liquid, patches, paste, cataplasm or sustained-release preparation, controlled-release preparation and the like.

The pharmaceutical dosage form of the pharmaceutical composition of the present invention may be a liquid dosage form, a solid dosage form, or a semisolid dosage form. The liquid dosage forms can be solution (including true solution and colloidal solution), emulsion (including o/w type, w/o type and multiple emulsion), suspension, injection (including water injection, powder injection and infusion), eye drop, nose drop, lotion, oral liquid, liniment, etc.; the solid dosage form can be tablet (including common tablet, enteric coated tablet, buccal tablet, dispersible tablet, chewable tablet, effervescent tablet, orally disintegrating tablet, sustained release tablet, controlled release tablet), capsule (including hard capsule, soft capsule, enteric coated capsule, sustained release capsule, controlled release capsule), granule, powder, pellet, dripping pill, suppository, pellicle, patch, aerosol (powder), spray, etc.; semisolid dosage forms can be ointments, gels, pastes, and the like.

The pharmaceutical composition can be prepared into common preparations, sustained release preparations, controlled release preparations, targeting preparations and various particle delivery systems.

For tableting the pharmaceutical composition of the present invention, a wide variety of excipients known in the art may be used, including diluents, binders, wetting agents, disintegrants, lubricants, glidants. The diluent can be starch, dextrin, sucrose, glucose, lactose, mannitol, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, calcium hydrogen phosphate, calcium carbonate, etc.; the humectant can be water, ethanol, isopropanol, etc.; the binder can be starch slurry, dextrin, syrup, Mel, glucose solution, microcrystalline cellulose, acacia slurry, gelatin slurry, sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; the disintegrant may be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, crosslinked polyvinylpyrrolidone, crosslinked sodium carboxymethylcellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfate, etc.; the lubricant and glidant may be talc, silicon dioxide, stearate, tartaric acid, liquid paraffin, polyethylene glycol, and the like. The tablets may be further formulated into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or double-layer and multi-layer tablets.

In order to prepare the pharmaceutical composition of the present invention into a capsule, the pharmaceutical composition of the present invention may be mixed with a diluent and a glidant, and the mixture may be directly placed into a hard capsule or a soft capsule. The pharmaceutical composition can also be prepared into granules or pellets with a diluent, an adhesive and a disintegrating agent, and then placed into hard capsules or soft capsules.

In order to prepare the pharmaceutical composition of the invention into injection, water, ethanol, isopropanol, propylene glycol, polyethylene glycol or a mixture thereof can be used as a solvent, and a proper amount of solubilizer, cosolvent, pH regulator and osmotic pressure regulator which are commonly used in the field can be added. The solubilizer or cosolvent can be ethanol, isopropanol, propylene glycol, polyethylene glycol, poloxamer, lecithin, hydroxypropyl-beta-cyclodextrin, etc.; the pH regulator can be citrate, phosphate, carbonate, acetate, hydrochloric acid, hydroxide, etc.; the osmotic pressure regulator can be sodium chloride, mannitol, glucose, phosphate, citrate, acetate, etc. For example, mannitol and glucose can be added as proppant for preparing lyophilized powder for injection.

In addition, colorants, preservatives, flavors, or other additives may also be added to the pharmaceutical preparation, if desired.

For the purpose of administration and enhancing the therapeutic effect, the drug or the pharmaceutical composition or the pharmaceutical preparation of the present invention can be administered by any known administration method.

The dosage of the pharmaceutical composition of the present invention to be administered may vary widely depending on the nature and severity of the disease to be prevented or treated, the individual condition of the patient or animal, the route of administration and the dosage form, and the like. Generally, the dosage range of the pharmaceutical composition of the present invention is 0.001-1000 mg/Kg body weight, preferably 0.01-500 mg/Kg body weight, and more preferably 0.1-200 mg/Kg body weight. The above-described dosage may be administered in one dosage unit or divided into several dosage units, depending on the clinical experience of the physician and the dosage regimen including the use of other therapeutic means.

The pharmaceutical composition can be taken alone or combined with other therapeutic drugs or symptomatic drugs. When the compound of the present invention is used in a synergistic manner with other therapeutic agents, the dosage thereof should be adjusted according to the actual circumstances.

The fourth purpose of the invention is realized by processing the Xuesaitong soft capsule preparation by taking the Xuesaitong medicinal composition as an effective component of a medicament and auxiliary materials and/or auxiliary agents.

Specifically, the soft capsule preparation for removing thromboembolism of the invention contains the following active pharmaceutical ingredients in percentage by mass:

ginsenoside Rb20.001-99.996%

Ginsenoside Rb10.001-99.996%

Ginsenoside Rg 10.001-99.996%

Ginsenoside Rc 0.001-99.996%

Notoginsenoside R10.001-99.996%

Preferably, the soft capsule preparation for removing the thrombus contains the following medicinal active ingredients in percentage by mass:

ginsenoside Rb20.01-99.96%

Ginsenoside Rb10.01-99.96%

Ginsenoside Rg 10.01-99.96%

Ginsenoside Rc 0.01-99.96%

10.01 to 99.96 percent of notoginsenoside R

Preferably, the soft capsule preparation for removing thromboembolism of the present invention contains the following pharmaceutical active ingredients by mass percent:

ginsenoside Rb254.6%

Ginsenoside Rb121.1%

Ginsenoside Rg 111.3%

Ginsenoside Rc 8.5%

Notoginsenoside R14.5%

Or

Ginsenoside Rb220.5%

Ginsenoside Rb147.8%

Ginsenoside Rg123.4%

Ginsenoside Rc 6.7%

Notoginsenoside R11.6%

Or

Ginsenoside Rb21.2%

Ginsenoside Rb114.6%

Ginsenoside Rg 155.7%

Ginsenoside Rc 19.3%

Notoginsenoside R19.2%

Or

Ginsenoside Rb221.1%

Ginsenoside Rb12.5%

Ginsenoside Rg 128.2%

Ginsenoside Rc 37.6%

Notoginsenoside R110.6%

Or

Ginsenoside Rb220.9%

Ginsenoside Rb12.7%

Ginsenoside Rg 121.5%

Ginsenoside Rc 10.2%

Notoginsenoside R144.7%

The fifth purpose of the invention is realized by weighing the pharmaceutical composition for treating thromboembolism as the active ingredient of the medicine, adding the pharmaceutical composition into the solvent, stirring the mixture evenly, putting the mixture into a pelleting press, pelleting the mixture with capsule skin and drying the mixture to obtain the medicine.

In the preparation method of the soft capsule for removing thromboembolism, the solvent is any one of water, ethanol, ethylene glycol, propylene glycol, glycerol, tween, sesame oil, peanut oil, ginger oil, zanthoxylum oil, rose oil, linseed oil, rapeseed oil, perilla oil, peony seed oil, black peach oil, sunflower oil, agastache oil, hemp seed oil, lard, beef tallow, mutton fat, fish oil, shark oil, snake oil and the like or a mixture of any two or more of the above in any proportion.

In the preparation method of the soft capsule for treating thromboembolism, the capsule skin is prepared by hot-melting, modulating, plastic and glue stabilizing colloid with a solvent and a blender. The colloid for preparing capsule skin is selected from animal gum such as gelatin, oxhide gum, dog skin gum, and pig skin gum, and also selected from vegetable gum such as carrageenan, guar gum, seaweed gum, locust bean gum, konjac gum, xanthan gum, etc. The solvent for preparing capsule shell is any one or mixture of any two or more of water, ethanol, propanol, ethylene glycol, isopropanol, n-glycerol, isobutanol, tert-butanol, n-butanol and polyethylene glycol at any ratio. The blending agents for preparing the capsule shell include toners such as lutein, carmine, amaranth, olive green, etc., also include acid modifiers such as citric acid, sodium citrate, carbonic acid, sodium carbonate, sodium bicarbonate, phosphoric acid, trisodium phosphate, disodium phosphate, monosodium phosphate, malic acid, sodium malate, lactic acid, sodium lactate, and also include flexibilizers such as cellulose, methyl cellulose, carboxymethyl cellulose, hydroxymethyl cellulose, beeswax, paraffin, stearic acid, glycerol monostearate, etc.

The sixth purpose of the invention is realized by the application of the pharmaceutical composition for treating the thromboembolism and/or the preparation thereof in preparing medicaments for preventing and/or treating cardiovascular and cerebrovascular diseases.

The pharmaceutical composition and/or the preparation thereof for treating the thromboembolism are applied to the preparation of medicines for preventing and/or treating ischemic cardiovascular and cerebrovascular diseases.

The pharmaceutical composition and/or the preparation thereof can be applied to the preparation of medicines for preventing and/or treating ischemic cerebral apoplexy, cerebral infarction and cerebral hemiplegia.

The pharmaceutical composition for treating thromboembolism and/or the preparation thereof are applied to preparation of drugs for preventing and/or treating retinopathy.

The pharmaceutical composition and/or the preparation thereof for treating the blood stasis are applied to the preparation of medicines for preventing and/or treating asthenopia or visual deterioration caused by optic atrophy, retinitis pigmentosa or macula retinae.

The pharmaceutical composition and/or the preparation thereof can be applied to the preparation of drugs for preventing and/or treating retinopathy caused by diabetes, cardiovascular diseases, cerebrovascular diseases and neurological diseases.

The invention takes the medicines such as the commercial panax notoginseng saponins and the like as the contrast, adopts the myocardial ischemia animal model of the SD rat, carries out the experimental study on the protective action of the pharmaceutical composition on the myocardial ischemia of the SD rat, and the result shows that: the pharmaceutical composition can obviously reduce the myocardial infarction area of a rat with myocardial ischemia, and has obvious protective effect on the myocardial ischemia; can obviously improve the cardiac function index of the rat with myocardial ischemia, and has obvious improvement effect on the cardiac dysfunction caused by myocardial ischemia injury; can obviously reduce the increase of serum LDH, CK, GOT and HBDH activity after coronary artery ligation of rats, and shows that the composition has protective effect on myocardial ischemia caused by coronary artery ligation. Experiments also show that the protective effect of the pharmaceutical composition on myocardial ischemia is better than that of contrast drugs such as commercial panax notoginseng saponins and the like with the same dose. The research shows that the pharmaceutical composition can be used for preventing or treating ischemic cardiovascular and cerebrovascular diseases, and the curative effect of the pharmaceutical composition is superior to that of the existing panax notoginseng saponins. See experimental example 1 for details.

The invention also takes the medicines such as the commercial panax notoginseng saponins and the like as the contrast, adopts an SD rat cerebral ischemia reperfusion injury animal model, carries out the experimental study on the protective effect of the pharmaceutical composition on the SD rat cerebral ischemia reperfusion injury, and the result shows that: the pharmaceutical composition can obviously improve the neurological symptoms of ischemia-reperfusion rats and has obvious treatment effect on the symptoms of stroke and hemiplegia caused by ischemia-reperfusion; the cerebral infarction volume of an ischemia-reperfusion rat can be remarkably reduced; the Evans blue content of the brain of the ischemia reperfusion rat can be obviously reduced, and the pharmaceutical composition has a protective effect on the blood brain barrier of the ischemia reperfusion rat. Experiments also show that the protective effect of the pharmaceutical composition on cerebral ischemia-reperfusion injury is better than that of commercial panax notoginseng saponins and other contrast drugs with the same dose. The research shows that the pharmaceutical composition can be used for preventing or treating ischemic cerebrovascular diseases such as ischemic cerebral apoplexy, cerebral infarction, cerebral hemiplegia and the like, and the curative effect of the pharmaceutical composition is superior to that of the existing panax notoginseng saponins. See experimental example 2 for details.

The invention also provides application of the pharmaceutical composition in preparing a medicament for preventing or treating retinopathy or optic nerve damage, in particular application of the pharmaceutical composition in preparing a medicament for preventing or treating asthenopia or visual deterioration caused by optic atrophy, retinitis pigmentosa or macula retinae and application of the pharmaceutical composition in preparing a medicament for preventing or treating retinopathy caused by diabetes, cardiovascular diseases, cerebrovascular diseases and neurological diseases.

The invention takes the medicines such as the commercial panax notoginseng saponins and the like as the contrast, adopts the SD rat persistent ocular hypertension retinal nerve injury animal model, carries out the experimental study on the protective action of the pharmaceutical composition on the SD rat retinal nerve injury, and the result shows that: the pharmaceutical composition can obviously reduce the damage of the persistent ocular hypertension to retinal nerves, and has good protection and repair effects on neuron damage. The experiment also shows that the protective effect of the pharmaceutical composition on the optic nerve injury is better than that of the contrast drugs such as the commercial panax notoginseng saponins and the like with the same dose. Therefore, the pharmaceutical composition can be used as a therapeutic drug for retina or optic neuropathy. See experimental example 3 for details.

Compared with the panax notoginseng saponins in the prior art, the pharmaceutical composition and preparation for treating thromboembolism provided by the invention have the following remarkable advantages and characteristics:

1. the pharmacological components are clear and definite, the components are fixed, and the product quality is uniform, stable and controllable.

2. The composition does not contain toxic and harmful non-pharmacological components or unknown components, has low toxic and side effects and adverse reactions, and has higher medication safety performance;

3. has definite pharmacological action and higher clinical curative effect.

The panax notoginseng saponins in the prior art contain more than 100 saponin components and a plurality of unknown components, have complicated and variable components, are difficult to control the quality, particularly have unclear action mechanisms, pharmacological actions and mutual interaction and influence of the components so far, have frequent adverse reactions, and bring a plurality of problems to the safety and the effectiveness of clinical medication. Compared with the prior art and products, the pharmaceutical composition and the preparation thereof for treating the thromboembolism have the advantages of higher curative effect, clear and fixed components, uniform and stable product quality, low toxic and side effects and adverse reactions, and greatly improved safety and effectiveness of medicament use.

Detailed Description

The present invention is further illustrated by the following examples, which are not intended to be limiting in any way, and any modifications or alterations based on the teachings of the present invention are intended to fall within the scope of the present invention.

The pharmaceutical composition for treating the thromboembolism comprises the following pharmaceutical components in percentage by mass:

ginsenoside Rb20.001-99.996%

Ginsenoside Rb10.001-99.996%

Ginsenoside Rg 10.001-99.996%

Ginsenoside Rc 0.001-99.996%

And (3) notoginsenoside R10.001-99.996%.

ginsenoside Rb20.01-99.96%

Ginsenoside Rb10.01-99.96%

Ginsenoside Rg 10.01-99.96%

Ginsenoside Rc 0.01-99.96%

10.01-99.96% of notoginsenoside R.

ginsenoside Rb254.6%

Ginsenoside Rb121.1%

Ginsenoside Rg 111.3%

Ginsenoside Rc 8.5%

Notoginsenoside R14.5%.

ginsenoside Rb220.5%

Ginsenoside Rb147.8%

Ginsenoside Rg123.4%

Ginsenoside Rc 6.7%

11.6 percent of notoginsenoside R.

ginsenoside Rb21.2%

Ginsenoside Rb114.6%

Ginsenoside Rg 155.7%

Ginsenoside Rc 19.3%

19.2 percent of notoginsenoside R.

ginsenoside Rb221.1%

Ginsenoside Rb12.5%

Ginsenoside Rg 128.2%

Ginsenoside Rc 37.6%

Notoginsenoside R110.6%.

ginsenoside Rb220.9%

Ginsenoside Rb12.7%

Ginsenoside Rg 121.5%

Ginsenoside Rc 10.2%

Notoginsenoside R144.7%.

The preparation method of the pharmaceutical composition for treating the thromboembolism comprises the following steps:

In the step 1), the mass ratio of the notoginsenoside R1 to the ethanol solvent is 0.05-5: 100, and the volume percentage concentration of the ethanol is 50-100%.

The mass ratio of the four ginsenosides in the step 2) to the active carbon for injection and the distilled water is 0.1-10: 0.01-0.5: 100.

the pre-freezing cooling rate in the step 3) is 5-15 ℃/h.

The pharmaceutical preparation for treating thromboembolism is prepared from the pharmaceutical composition for treating thromboembolism and pharmaceutical auxiliary materials, carriers and matrixes, and comprises injections, tablets, oral liquid, capsules, soft capsules, dropping pills, sustained-release preparations and controlled-release preparations.

The soft capsule preparation of the invention is prepared by taking the pharmaceutical composition of the Xuesaitong as the effective component of the medicine and assisting with auxiliary materials and/or auxiliary agents.

The soft capsule preparation for treating the thromboembolism comprises the following medicinal active ingredients in percentage by mass:

ginsenoside Rb20.001-99.996%

Ginsenoside Rb10.001-99.996%

Ginsenoside Rg 10.001-99.996%

Ginsenoside Rc 0.001-99.996%

And (3) notoginsenoside R10.001-99.996%.

ginsenoside Rb20.01-99.96%

Ginsenoside Rb10.01-99.96%

Ginsenoside Rg 10.01-99.96%

Ginsenoside Rc 0.01-99.96%

10.01-99.96% of notoginsenoside R.

ginsenoside Rb254.6%

Ginsenoside Rb121.1%

Ginsenoside Rg 111.3%

Ginsenoside Rc 8.5%

Notoginsenoside R14.5%.

ginsenoside Rb220.5%

Ginsenoside Rb147.8%

Ginsenoside Rg123.4%

Ginsenoside Rc 6.7%

11.6 percent of notoginsenoside R.

ginsenoside Rb21.2%

Ginsenoside Rb114.6%

Ginsenoside Rg 155.7%

Ginsenoside Rc 19.3%

19.2 percent of notoginsenoside R.

ginsenoside Rb221.1%

Ginsenoside Rb12.5%

Ginsenoside Rg 128.2%

Ginsenoside Rc 37.6%

Notoginsenoside R110.6%.

ginsenoside Rb220.9%

Ginsenoside Rb12.7%

Ginsenoside Rg 121.5%

Ginsenoside Rc 10.2%

Notoginsenoside R144.7%.

The preparation method of the soft capsule preparation for removing thromboembolism provided by the invention comprises the steps of weighing the pharmaceutical composition for removing thromboembolism as an active ingredient of a medicine, adding the pharmaceutical composition into a solvent, uniformly stirring, placing the mixture into a pill press, pressing pills with capsule shells, and drying to obtain the soft capsule preparation.

The solvent is any one or a mixture of more than two of water, ethanol, ethylene glycol, propylene glycol, glycerol, tween, peanut oil, ginger oil, zanthoxylum oil, rose oil, rapeseed oil, peony seed oil, black peach oil, sunflower oil, patchouli oil, lard, beef tallow, mutton fat, fish oil, shark oil and snake oil in any proportion.

The capsule skin is prepared by hot-melting, blending, plastic cement and stabilizing glue with a colloid, a solvent and a blender.

The colloid for preparing capsule shell is selected from animal gum or vegetable gum.

The animal glue comprises gelatin, oxhide glue, dog hide glue or pig hide glue.

The vegetable gum comprises carrageenan, guar gum, seaweed gum, locust bean gum, konjac gum and xanthan gum.

The solvent for preparing capsule shell is any one or mixture of any two or more of water, ethanol, propanol, ethylene glycol, isopropanol, n-glycerol, isobutanol, tert-butanol, n-butanol and polyethylene glycol at any ratio.

The preparation for capsule skin comprises toner and acid regulating agent.

The toner comprises carmine, amaranth or olive green.

The acid regulator comprises citric acid, sodium citrate, carbonic acid, sodium carbonate, sodium bicarbonate, phosphoric acid, trisodium phosphate, disodium phosphate, monosodium phosphate, malic acid, sodium malate, lactic acid or sodium lactate.

The preparation for capsule skin also comprises a softening agent.

The softening agent comprises cellulose, methylcellulose, carboxymethyl cellulose, hydroxymethyl cellulose, beeswax, paraffin, stearic acid or glycerol monostearate.

The application of the pharmaceutical composition and/or the preparation thereof for treating the thromboembolism is the application of the pharmaceutical composition and/or the preparation thereof for preparing the medicines for preventing and/or treating the cardiovascular and cerebrovascular diseases.

The invention is further illustrated below with reference to specific embodiments:

EXAMPLE 11 preparation of pharmaceutical composition #

The raw materials comprise:

ginsenoside Rb254.6 g

Ginsenoside Rb121.1 g

Ginsenoside Rg 111.3 g

Ginsenoside Rc 8.5 g

Notoginsenoside R14.5g

In total 100g

The preparation method comprises the following steps:

1) weighing 14.5 g of notoginsenoside R, placing in 100g of ethanol solvent with the volume percentage concentration of 50%, and stirring for dissolving to obtain a solution A;

2) weighing ginsenoside Rb254.6 g, ginsenoside Rb121.1 g, ginsenoside Rg111.3 g and ginsenoside Rc 8.5 g, dissolving in 900 g distilled water under stirring, heating to 50-70 deg.C, adding solution A under stirring, adding 2.0g of active carbon for injection, heating to boil for 30-50 min, standing, cooling to room temperature, and filtering to remove active carbon;

3) and (3) placing the filtrate in a vacuum freeze dryer, cooling to minus 40 to minus 30 ℃ at a cooling rate of 5 ℃/h, pre-freezing for 3-5 hours, controlling the temperature to minus 25 to minus 20 ℃ and the pressure to 10-20 Pa, carrying out sublimation drying for 3-5 hours, controlling the temperature to 30-50 ℃ and the pressure to 1-10 Pa, carrying out resolution drying for 4-6 hours, and recovering to normal pressure room temperature to obtain the product.

EXAMPLE 22 preparation of pharmaceutical composition

The raw materials comprise:

ginsenoside Rb220.5 g

Ginsenoside Rb147.8 g

Ginsenoside Rg123.4 g

Ginsenoside Rc 6.7 g

Notoginsenoside R11.6 g

In total 100g

The preparation method comprises the following steps:

1) weighing 11.6 g of notoginsenoside R, placing in 100g of ethanol solvent with the volume percentage concentration of 80%, and stirring for dissolving to obtain solution A;

2) weighing ginsenoside Rb220.5 g, ginsenoside Rb147.8 g, ginsenoside Rg123.4 g and ginsenoside Rc 6.7 g, dissolving in 950 g distilled water under stirring, heating to 50-70 deg.C, adding solution A under stirring, adding activated carbon for injection 1.0g, heating to boil for 30-50 min, standing, cooling to room temperature, and filtering to remove activated carbon;

3) and (3) placing the filtrate in a vacuum freeze dryer, cooling to minus 40 to minus 30 ℃ at a cooling rate of 15 ℃/h, pre-freezing for 3-5 hours, controlling the temperature to minus 25 to minus 20 ℃ and the pressure to 10-20 Pa, carrying out sublimation drying for 3-5 hours, controlling the temperature to 30-50 ℃ and the pressure to 1-10 Pa, carrying out resolution drying for 4-6 hours, and recovering to normal pressure room temperature to obtain the product.

EXAMPLE 33 preparation of pharmaceutical composition

The raw materials comprise:

ginsenoside Rb21.2 g

Ginsenoside Rb114.6 g

Ginsenoside Rg 155.7 g

Ginsenoside Rc 19.3 g

Notoginsenoside R19.2 g

In total 100g

The preparation method comprises the following steps:

1) weighing 19.2 g of notoginsenoside R, placing in 500 g of ethanol solvent with volume percentage concentration of 100%, and stirring for dissolving to obtain solution A;

2) weighing ginsenoside Rb21.2 g, ginsenoside Rb114.6 g, ginsenoside Rg155.7 g and ginsenoside Rc 19.3 g, dissolving in 1000 g distilled water under stirring, heating to 50-70 deg.C, adding solution A under stirring, adding activated carbon for injection 3.0g, heating to boil for 30-50 min, standing, cooling to room temperature, and filtering to remove activated carbon;

3) and (3) placing the filtrate in a vacuum freeze dryer, cooling to minus 40 to minus 30 ℃ at a cooling rate of 10 ℃/h, pre-freezing for 3-5 hours, controlling the temperature to minus 25 to minus 20 ℃ and the pressure to 10-20 Pa, carrying out sublimation drying for 3-5 hours, controlling the temperature to 30-50 ℃ and the pressure to 1-10 Pa, carrying out resolution drying for 4-6 hours, and recovering to normal pressure room temperature to obtain the product.

EXAMPLE 44 preparation of pharmaceutical composition

The raw materials comprise:

ginsenoside Rb221.1 g

Ginsenoside Rb12.5g

Ginsenoside Rg 128.2g

Ginsenoside Rc 37.6g

Notoginseng radix Saponin R110.6g

In total 100g

The preparation method comprises the following steps:

1) weighing 110.6g of notoginsenoside, placing in 1500 g of ethanol solvent with volume percentage concentration of 60%, and stirring for dissolving to obtain solution A;

2) weighing ginsenoside Rb221.1 g, ginsenoside Rb12.5 g, ginsenoside Rg128.2g and ginsenoside Rc 37.6g, dissolving in 5000 g of distilled water under stirring, heating to 50-70 deg.C, adding solution A under stirring, adding 8.0g of active carbon for injection, heating to boil for 30-50 min, standing, cooling to room temperature, and filtering to remove active carbon;

3) and (3) putting the filtrate into a vacuum freeze dryer, cooling to minus 40 to minus 30 ℃ at a cooling rate of 12 ℃/h, pre-freezing for 3 to 5 hours, controlling the temperature to minus 25 to minus 20 ℃ and the pressure to 10 to 20Pa, carrying out sublimation drying for 3 to 5 hours, controlling the temperature to 30 to 50 ℃ and the pressure to 1 to 10Pa, carrying out resolution drying for 4 to 6 hours, and recovering to the normal-pressure room temperature to obtain the product.

EXAMPLE 55 preparation of pharmaceutical composition

The raw materials comprise:

ginsenoside Rb220.9 g

Ginsenoside Rb12.7g

Ginsenoside Rg 121.5g

Ginsenoside Rc 10.2g

Notoginseng radix Saponin R144.7g

In total 100g

The preparation method comprises the following steps:

1) weighing 144.7g of notoginsenoside, placing in 4500 g of 90% ethanol solvent by volume percent, stirring and dissolving to obtain solution A;

2) weighing ginsenoside Rb220.9 g, ginsenoside Rb12.7 g, ginsenoside Rg121.5g and ginsenoside Rc 10.2g, dissolving in 3000 g distilled water under stirring, heating to 50-70 deg.C, adding solution A under stirring, adding activated carbon for injection 1.0g, heating to boil for 30-50 min, standing, cooling to room temperature, and filtering to remove activated carbon;

EXAMPLE 61 preparation of # control sample

The raw materials comprise:

ginsenoside Rb 150g

Ginsenoside Rg 150g

In total 100g

The preparation method comprises the following steps:

weighing ginsenoside Rb 150g and ginsenoside Rg 150g, putting the materials into 1000 g of 75% ethanol solvent by volume percentage, stirring and dissolving, adding 1.5g of activated carbon for injection, heating and boiling for 30-50 minutes, standing and cooling to room temperature, filtering to remove the activated carbon, putting the filtrate into a vacuum freeze dryer, cooling to-40 to-30 ℃ at a cooling rate of 15 ℃/h, pre-freezing for 3-5 hours, controlling the temperature to be-25 to-20 ℃, the pressure to be 10-20 Pa, carrying out sublimation drying for 3-5 hours, controlling the temperature to be 30-50 ℃, the pressure to be 1-10 Pa, carrying out resolution drying for 4-6 hours, and recovering to normal pressure and room temperature to obtain the ginsenoside Rb-Rg-Rb-Rg.

EXAMPLE 72 preparation of # control sample

The raw materials comprise:

notoginseng radix saponin R130 g

Ginsenoside Rb 135 g

Ginsenoside Rg 135 g

In total 100g

The preparation method comprises the following steps:

weighing notoginsenoside R130 g, ginsenoside Rb 135 g and ginsenoside Rg 135 g, putting the notoginsenoside R130 g, ginsenoside Rb 135 g and ginsenoside Rg 135 g into 1000 g of 75% ethanol solvent by volume percentage, stirring and dissolving, adding 1.5g of activated carbon for injection, heating and boiling for 30-50 minutes, standing and cooling to room temperature, filtering to remove the activated carbon, putting the filtrate into a vacuum freeze dryer, cooling to-40 to-30 ℃ at a cooling rate of 15 ℃/h, pre-freezing for 3-5 hours, then controlling the temperature to-25 to-20 ℃, controlling the pressure to 10-20 Pa, carrying out sublimation drying for 3-5 hours, then controlling the temperature to 30-50 ℃, the pressure to 1-10 Pa, carrying out resolution drying for 4-6 hours, and recovering to the room temperature under normal pressure to obtain the oral liquid.

EXAMPLE 83 preparation of # control sample

The raw materials comprise:

ginsenoside Rb 225 g

Notoginseng radix saponin R125 g

Ginsenoside Rb 125 g

Ginsenoside Rg 125 g

In total 100g

The preparation method comprises the following steps:

weighing 225 g of ginsenoside Rb, 125 g g of notoginsenoside, 125 g of ginsenoside Rb and 125 g, placing the materials in 1000 g of 75 volume percent ethanol solvent, stirring for dissolving, adding 1.5g of active carbon for injection, heating and boiling for 30-50 minutes, standing and cooling to room temperature, filtering to remove the active carbon, placing the filtrate in a vacuum freeze dryer, cooling to-40 to-30 ℃ at a cooling rate of 15 ℃/h, pre-freezing for 3-5 hours, controlling the temperature to-25 to-20 ℃, the pressure to 10-20 Pa, carrying out sublimation drying for 3-5 hours, controlling the temperature to 30-50 ℃ and the pressure to 1-10 Pa, carrying out resolution drying for 4-6 hours, and recovering to the room temperature under normal pressure to obtain the ginsenoside Rb-125-g.

EXAMPLE 9 tablets

The medicine prescription (mass percent):

the No. 1 pharmaceutical composition of the invention example 1 is 15.0%

62.5 percent of starch

Microcrystalline cellulose 13.2%

Sodium carboxymethyl starch 9.3%

The total is 100 percent

According to the formula ratio, the No. 1 pharmaceutical composition provided by the invention is mixed with starch, microcrystalline cellulose and sodium carboxymethyl starch, and then the mixture is tabletted on a tabletting machine to obtain the compound tablet.

EXAMPLE 10 capsules

The medicine prescription (mass percent):

the 2# pharmaceutical composition of the invention example 2 is 10.0%

69.0 percent of starch

Microcrystalline cellulose 12.5%

Magnesium stearate 3.2%

Sodium carboxymethyl starch 5.3%

The total is 100 percent

According to the formula proportion, the 2# pharmaceutical composition of the invention in the embodiment 2 is mixed with starch, microcrystalline cellulose, sodium carboxymethyl starch and magnesium stearate and then filled into hard gelatin capsules to obtain the pharmaceutical composition.

Example 11 oral liquid

The medicine prescription (mass percent):

5.2% of the No. 3 pharmaceutical composition of example 3 of the invention

12.3 percent of sucrose

0.5 percent of ethanol

82.0 percent of distilled water

The total is 100 percent

Mixing and dissolving the 3# pharmaceutical composition of the embodiment 3 of the invention with cane sugar, ethanol and distilled water according to the proportion of the prescription, filtering, encapsulating the mixture in 10 mL/capsule in oral liquid, and carrying out damp-heat sterilization at 100 ℃ for 30min to obtain the oral liquid.

EXAMPLE 12 injection solution

The prescription of the medicine is as follows:

55g of No. 4 pharmaceutical composition of example 4 of the present invention

90g of sodium chloride

Proper amount of water for injection

Making into 1000ml

According to the formula, the No. 4 pharmaceutical composition provided by the invention is added with sodium chloride and a proper amount of water for injection, the mixture is stirred uniformly, 0.1% of activated carbon for injection is added, the mixture is adsorbed, filtered and decarbonized, the water for injection is added to a specified amount, the mixture is filtered by a microporous filter membrane, the mixture is encapsulated according to 5 mL/bag, and the mixture is sterilized by moist heat at 100 ℃ for 30min and qualified through light inspection, thus obtaining the pharmaceutical composition.

EXAMPLE 13 lyophilized powder for injection

The prescription of the medicine is as follows:

100g of 5# pharmaceutical composition of example 5 of the present invention

90g of sodium chloride

Mannitol 75g

Proper amount of water for injection

Making into 1000ml

According to the formula proportion, the 5# pharmaceutical composition provided by the invention and prepared by the preparation method is added with sodium chloride, mannitol and a proper amount of water for injection, the mixture is uniformly stirred, 0.1% of activated carbon for injection is added, the mixture is adsorbed, filtered and decarbonized, the water for injection is supplemented to a specified amount, the mixture is filtered by a microporous filter membrane, the mixture is subpackaged by 2 mL/piece, and the product is obtained after freeze drying, packaging and passing inspection.

Example 141 preparation of XUESAITONG Soft Capsule

The prescription of the medicine is as follows:

100g of No. 1 pharmaceutical composition of example 1 of the present invention

No. 1 solvent 250g

No. 1 capsule skin 100g

Making into 1000 soft capsules

The preparation method comprises the following steps:

weighing 100g of the No. 1 pharmaceutical composition of the embodiment 1, adding 250g of the No. 1 solvent, uniformly stirring, putting into a pelleting press, pelleting with No. 1 capsule skin, and drying to obtain each Xuesaitong soft capsule containing 100mg of the active ingredient.

Solvent composition No. 1 (mass percent)

48.5 percent of normal glycerol

20.0 percent of ethanol

25.8 percent of water

0.1 percent of rose oil

5.6 percent of black peach oil

The total is 100 percent

Preparation method of No. 1 solvent:

weighing n-glycerol, ethanol, water, rose oil and black peach oil according to the formula proportion, mixing, and stirring uniformly to obtain the product.

The 1# capsule skin formula (mass percent):

18.2 percent of gelatin

75.9 percent of water

0.2 percent of citric acid

Monostearyl fatty acid glyceride 0.5%

5.2 percent of carboxymethyl cellulose

The total is 100 percent

The preparation method of the No. 1 capsule shell comprises the following steps: weighing gelatin according to the formula ratio, adding the gelatin into water, heating and stirring at 60-80 ℃ for dissolving, then sequentially adding carboxymethyl cellulose, glycerol monostearate and citric acid for modulation and plastic cement, and stabilizing the glue at 30-40 ℃ for 24 hours to obtain the gelatin.

EXAMPLE 152 preparation of XUESAITONG Soft Capsule

The prescription of the medicine is as follows:

120g of No. 2 pharmaceutical composition of example 2 of the present invention

350g of No. 2 solvent

2# Capsule skin 150g

Making into 2000 soft capsules

The preparation method comprises the following steps:

weighing 120g of the No. 2 pharmaceutical composition of the embodiment 2, adding 350g of the No. 2 solvent, uniformly stirring, putting into a pelleting press, pelleting with No. 2 capsule skin, and drying to obtain each Xuesaitong soft capsule containing 60mg of the effective component.

2# solvent composition (mass percentage)

42.3 percent of ethylene glycol

15.2 percent of ethanol

18.6 percent of water

Tween 602.8%

Peanut oil 21.1%

The total is 100 percent

Preparation method of No. 2 solvent:

according to the formula proportion, the glycol, the ethanol, the water, the Tween 60 and the peanut oil are weighed, mixed and stirred uniformly to obtain the peanut oil.

The 2# capsule skin formula (mass percent):

seaweed gel 23.2%

70.4 percent of water

0.02 percent of amaranth

Disodium phosphate 0.87%

0.21 percent of monosodium phosphate

5.3 percent of methyl cellulose

The total is 100 percent

2# Capsule skin preparation method: weighing seaweed gel according to the formula ratio, adding the seaweed gel into water, heating and stirring at 60-80 ℃ for dissolving, then sequentially adding methyl cellulose, amaranth, disodium phosphate and monosodium phosphate for blending and plastic cement, and stabilizing the glue at 30-40 ℃ for 24 hours to obtain the seaweed gel.

Example 163 preparation of XUESAITONG Soft Capsule

The prescription of the medicine is as follows:

60g of No. 3 pharmaceutical composition of example 3 of the present invention

400g of No. 3 solvent

No. 3 capsule shell 200 g

Making into 2000 soft capsules

The preparation method comprises the following steps:

weighing 60g of the 3# pharmaceutical composition of the embodiment 3, adding 400g of the 3# solvent, uniformly stirring, putting into a pelleting press, pelleting with 3# capsule skin, and drying to obtain each Xuesaitong soft capsule containing 30mg of the active ingredient.

3# solvent composition (mass percentage)

35.3 percent of propylene glycol

Polyethylene glycol-40026.5%

Tween 606.8%

31.4 percent of fish oil

The total is 100 percent

Preparation method of No. 3 solvent:

weighing propylene glycol, polyethylene glycol-400, tween 60 and fish oil according to the formula proportion, mixing, and stirring uniformly to obtain the composition.

The 3# capsule skin formula (mass percent):

guar gum 16.3%

Xanthan gum 7.5%

60.4 percent of water

6.2 percent of ethylene glycol

Sodium lactate 1.3%

4.8 percent of beeswax

Hydroxymethyl cellulose 3.5%

The total is 100 percent

The preparation method of the 3# capsule shell comprises the following steps: weighing guar gum, xanthan gum and ethylene glycol according to the formula ratio, adding into water, heating and stirring at 60-80 ℃ for dissolving, then sequentially adding hydroxymethyl cellulose, beeswax and sodium lactate for blending, and plastic cement, and stabilizing the rubber at 30-40 ℃ for 24 hours to obtain the guar gum.

Example 174 preparation of XUESAITONG Soft Capsule

The prescription of the medicine is as follows:

50g of No. 4 pharmaceutical composition of example 4 of the present invention

Solvent No. 4 (300 g)

No. 4 capsule skin 150g

Making into 2000 soft capsules

The preparation method comprises the following steps:

weighing 50g of the No. 4 pharmaceutical composition of the embodiment 4, adding 300g of the No. 4 solvent, uniformly stirring, putting into a pelleting press, pelleting with No. 4 capsule skin, and drying to obtain the Xuesaitong soft capsule containing 25mg of the active ingredient in each capsule.

4# solvent composition (mass percentage)

79.3 percent of water

20.7 percent of normal glycerol

The total is 100 percent

Preparation method of No. 4 solvent:

weighing water and normal glycerol according to the formula ratio, mixing, and stirring uniformly to obtain the product.

The 4# capsule skin formula (mass percent):

gelatin 12.5%

6.9 percent of carrageenan

65.4 percent of water

Polyethylene glycol-4009.7%

0.9 percent of sodium bicarbonate

1.5 percent of paraffin

3.1 percent of cellulose

The total is 100 percent

The preparation method of the No. 4 capsule shell comprises the following steps: weighing gelatin, carrageenan and polyethylene glycol-400 according to the formula proportion, adding into water, heating and stirring at 60-80 ℃ for dissolving, then sequentially adding cellulose, paraffin and sodium bicarbonate for blending and plastic cement, and stabilizing the glue at 30-40 ℃ for 24 hours to obtain the gelatin.

Example 185 preparation of XUESAITONG Soft Capsule

The prescription of the medicine is as follows:

150g of 5# pharmaceutical composition of example 5 of the present invention

400g of No. 5 solvent

No. 5 capsule shell 200 g

Making into 1000 soft capsules

The preparation method comprises the following steps:

weighing 150g of the 5# pharmaceutical composition of the embodiment 5, adding 400g of the 5# solvent, uniformly stirring, putting into a pelleting press, pelleting with 5# capsule skin, and drying to obtain each Xuesaitong soft capsule containing 150mg of the active ingredient.

5# solvent composition (mass percentage)

58.1 percent of ethanol

27.6 percent of water

Ethylene glycol 14.3%

The total is 100 percent

Preparation method of No. 5 solvent:

according to the formula proportion, the ethanol, the water and the ethylene glycol are weighed, mixed and stirred uniformly to obtain the water-based paint.

The 5# capsule skin formula (mass percent):

5.3 percent of oxhide glue

Locust bean gum 13.7%

75.4 percent of water

Olive green 0.03%

0.97 percent of sodium malate

Monostearyl fatty acid glyceride 1.5%

3.1 percent of methyl cellulose

The total is 100 percent

The preparation method of the No. 5 capsule shell comprises the following steps: weighing the oxhide gelatin and the locust bean gum according to the formula ratio, adding the oxhide gelatin and the locust bean gum into water, heating and stirring the mixture at the temperature of between 60 and 80 ℃ to dissolve the mixture, then sequentially adding the olive green, the methyl cellulose, the monostearyl fatty acid glyceride and the sodium malate to blend and mold the mixture, and stabilizing the mixture at the temperature of between 30 and 40 ℃ for 24 hours to obtain the artificial leather.

Experimental example 1 protective action of the pharmaceutical composition of the present invention on myocardial ischemia in SD rats

1. Materials and methods

1.1 drugs and reagents

1# pharmaceutical composition: ginsenoside Rb254.6%, ginsenoside Rb121.1%, ginsenoside Rg111.3%, ginsenoside Rc 8.5%, and notoginsenoside R14.5%, by making use of example 1.

2# pharmaceutical composition: ginsenoside Rb220.5%, ginsenoside Rb147.8%, ginsenoside Rg123.4%, ginsenoside Rc 6.7%, and notoginsenoside R11.6%, by preparing as in example 2.

3# pharmaceutical composition: ginsenoside Rb21.2%, ginsenoside Rb114.6%, ginsenoside Rg155.7%, ginsenoside Rc 19.3%, and notoginsenoside R19.2%, by weight, as in example 3.

4# pharmaceutical composition: ginsenoside Rb221.1%, ginsenoside Rb12.5%, ginsenoside Rg128.2%, ginsenoside Rc 37.6%, and notoginsenoside R110.6%, by making use of example 4.

5# pharmaceutical composition: ginsenoside Rb220.9%, ginsenoside Rb12.7%, ginsenoside Rg121.5%, ginsenoside Rc 10.2%, and notoginsenoside R144.7%, by weight, as in example 5.

Control sample # 1: ginsenoside Rb 150% and ginsenoside Rg 150% were prepared as in example 6.

Control sample # 2: ginsenoside Rb 135%, ginsenoside Rg 135%, and notoginsenoside R130%, prepared as in example 7.

Control sample # 3: ginsenoside Rb 125%, ginsenoside Rg 125%, notoginsenoside R125%, and ginsenoside Rb 225% were prepared as in example 8.

Commercially available panax notoginseng saponins: the quality of the product meets the requirements of the 2015 edition of Chinese pharmacopoeia, and is purchased from Chinese medicine GmbH.

Triphenyltetrazolium Chloride (TTC): supplied by fine chemicals, Inc., of Ma, Beijing.

Sodium pentobarbital: supplied by Beijing Chemicals, Inc.

Heparin sodium injection: offered by Wanbang, Jiangsu, Biochemical medicine, Inc.

Creatine Kinase (CK) kit and Lactate Dehydrogenase (LDH) kit: supplied by Beijing Zhongsheng Beizhong Beijing Biotechnology GmbH.

1.2 Main instruments

HX-300 animal respirator and BL-420S biological function appearance: chengduta alliance technologies, Inc.;

TGL-16G-A high speed refrigerated centrifuge: shanghai' an pavilion scientific medical instrument factory;

hitachi 7600 full-automatic biochemical analyzer: shimadzu corporation, Japan;

BS 110S electronic balance: sartorius, germany.

1.3 animal grouping and administration

SD male rats 110, with a mass of 250 ± 30g, were randomly divided into 11 groups: the test results show that the.

The rats in each drug group are respectively given 50mg of ig^-1The sham-operated group and the model group were administered with the same amount of the solvent 2 times a day for 7 days. After the last administration for 30min, ligation of anterior descending coronary artery was performed to prepare an acute myocardial ischemia model.

1.4 preparation of acute myocardial ischemia animal model

Rats were ip given 60mg of 1% sodium pentobarbital^-1Anaesthetizing, connecting electrocardiogram electrodes under the skin of four limbs, and recording standard II-lead electrocardiogram. Inserting trachea through mouth and connecting with animal respirator. The chest is opened between the fourth intercostal on the left side, the heart is squeezed out slightly, the lower edge of the junction of the pulmonary artery cone and the left auricle is ligated by 1-2 mm by silk thread, the heart is rapidly put back into the chest cavity, and the ligation part is cyanotic, bulging and ST segment lifting of electrocardiogram are taken as signs of successful ligation. The false operation group only perforates the needle at the corresponding position of the left anterior descending branch without knotting, and the rest operations are the same as the operation group. Observing for 10 min after operation, squeezing out air in thoracic cavity and closing thoracic layer by layer, recovering spontaneous respiration of rat, pulling out trachea cannula and removing tracheal endocrine, and feeding with feed and water for 24 hr. In the experimental process, all rats which die due to surgery, anesthesia accident and the like and have postoperative survival time which does not meet the requirement of material drawing time are not listed in experimental observation items.

1.5 cardiac function testing

60 mg/kg of 1% sodium pentobarbital is given in ip 24 hours after the rat ligates the left anterior descending branch of the coronary artery^－1Anaesthesia, fixing in the supine position, cutting the skin in the middle of the neck, and blunt-separating the neck muscles with hemostats to expose the right common carotid artery. The proximal end is threaded with silk thread, the distal end is clamped with artery clamp, a V-shaped incision is cut on the right common carotid artery, and the insertion is filled with 250 kU.L^－1A catheter of heparin in saline solution and fixed with silk thread. Loosening the artery clamp toThe heparinized PE catheter is inserted into the left ventricle through the right common carotid artery, and whether the insertion tube enters the ventricle cavity or not is judged according to the change of the pressure graph shown by the display. The other end of the conduit is connected with a pressure transducer, and signals are input into a BL-420S biological signal acquisition and analysis system to monitor the hemodynamics change of the left ventricle. After the left ventricle is inserted and balanced for 30min, the Left Ventricular Systolic Pressure (LVSP) and the Left Ventricular End Diastolic Pressure (LVEDP) are recorded, and simultaneously, the left ventricular pressure electric signal is synchronously input into a differentiator, and the maximum rising rate (+ dp/dt max) of the left ventricular pressure in the isovolumetric systolic period and the maximum falling rate (-dp/dt max) of the left ventricular pressure in the isovolumetric diastolic period are measured.

1.6 TTC staining for myocardial infarction area

After the cardiac function is detected, the heart is taken out quickly, washed in ice-cold normal saline to remove blood stain, and non-myocardial tissues such as the atrium, blood vessels, fat and the like are removed, and the heart is placed in a refrigerator at the temperature of minus 20 ℃ for 30 min. Under the cardiac ligature, the heart was cut into 5 pieces (1.5-2 mm thick) from the apex to the bottom of the heart in parallel coronary sulci, and incubated in 1% TTC phosphate buffer solution (pH 7.4) in a thermostatic water bath at 37 ℃ for 10 min. Fixing the stained heart transverse section in 4% paraformaldehyde solution, scanning the front and back sides of the heart section, calculating the infarction range by Photoshop software, and expressing the infarction area as the percentage of the whole ventricular area.

1.7 detection of serum myocardial enzymes

After the cardiac function test is finished, the abdominal aorta is bled, centrifuged at 1200 r/min and 4 ℃ for 15min to obtain a serum sample, and the serum sample is frozen in a refrigerator at minus 80 ℃ for later use. The serum myocardial zymogram detection comprises CK, LDH, alpha-hydroxybutyrate dehydrogenase (alpha-HBDH), glutamic-pyruvic transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT), and is performed by adopting a Hitachi 7600 full-automatic biochemical analyzer for determination.

2. Results of the experiment

2.1 Effect on myocardial ischemia damaged rat infarct size

The experimental results show that the pharmaceutical composition can significantly reduce the myocardial infarction area of rats with myocardial ischemia, has obvious protective effect on myocardial ischemia, and has better effect than 1-3# control samples and commercial panax notoginseng saponins control drugs.

2.2 Effect on cardiac function in rats with myocardial ischemia

The experimental results show that the pharmaceutical composition can obviously improve the cardiac function index of a myocardial ischemia rat, has obvious improvement effect on cardiac dysfunction caused by myocardial ischemia injury, and has better effect than a 1-3# control sample and a commercial panax notoginseng saponins control medicament.

2.3 Effect on serum myocardial enzymes in rats with myocardial ischemia

The experimental results show that the pharmaceutical composition can obviously reduce the increase of serum LDH, CK, GOT and HBDH activity after rat coronary artery ligation, and the pharmaceutical composition has a protective effect on myocardial ischemia caused by coronary artery ligation, and the effect is superior to that of a 1-3# control sample and a commercially available panax notoginseng saponins control medicament.

And (4) conclusion: the pharmaceutical composition can obviously reduce the myocardial infarction range of a rat after coronary artery ligation, improve the cardiac function of the rat after the myocardial infarction, and reduce the increase of serum LDH, CK, GOT and HBDH activity of the rat after coronary artery ligation, which shows that the pharmaceutical composition has a protective effect on myocardial ischemia caused by coronary artery ligation, and the effect of the pharmaceutical composition is superior to that of a control sample and commercially available panax notoginseng saponins.

Experimental example 2 study on protective effect of cerebral ischemia reperfusion injury in rat

1. Materials and instruments

1.1 drugs and reagents

The above test medicines are mixed with 0.9% normal saline to obtain 1.0% solution.

Interleukin-beta (IL-1 beta), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), interleukin-8 (IL-8) ABC-ELISA test kit: supplied by Sigma company.

Evans Blue (Evans Blue): fluka corporation;

formamide: tianjin outer ring chemical company;

2% tetrazolium red (TTC): supplied by Sigma company.

1.2 Main instruments

CR-412 Low temperature Normal speed centrifuge: supplied by Joean, France;

721, spectrophotometer: the third instrument factory in Shanghai;

computer image analysis system: produced by Shanghai medical university.

2. Experimental methods

2.1 establishment of cerebral ischemia-reperfusion model

SD rats 330, randomly divided into 11 groups: the test results show that the. Model group and each drug group rats ip 10% chloral hydrate 3.5 mL^{－ 1}Anaesthetizing, fixing on back, separating left Common Carotid Artery (CCA), Internal Carotid Artery (ICA) and External Carotid Artery (ECA), ligating ECA and CCA, clamping ICA distal end with artery clamp, making an incision at common carotid artery about 0.5 cm away from ECA and ICA bifurcation rapidly, inserting a round and blunt nylon thread with diameter of 0.3 mm at one end, inserting depth (20.0 +/-2.0 mm, realizing cerebral ischemia caused by middle cerebral artery occlusion, reserving 1 cm outside the nylon thread, suturing skin, slightly lifting the reserved thread after 2 h to have slight resistance to realize middle cerebral artery reperfusion.

2.2 animal dosing and treatment

The medicine groups are administrated once in ip 15min before ischemia and 6 h after ischemia, and the dosage of each administration is 10 mg/kg^{－ 1}. The sham operation group and the model group were given the same amount of physiological saline ip.

2.3 neurobehavioral Scoring

After recovery from anesthesia, animals were returned to their cages, free access, and neuro-behavioral scoring was performed 24 h post-surgery. Scoring with reference to the 5-point standard score of Zea Longa: the symptom of no nerve injury was 0 point, the healthy side forepaw could not be fully extended for 1 point, the healthy side was turned for 2 points, the healthy side was tilted for 3 points, the spontaneous walking was not possible, and the consciousness loss was 4 points.

2.4 infarct volume determination

10 rats in each group were taken, the brains were cut off after 2 h reperfusion for 22 h after cerebral ischemia, and the brains were coronal-dissected at 1, 3, 5, 7 mm from the forehead with a brain dissector according to the thickness of 2 mm. The brain slices are immersed in 2% tetrazolium red at 37 ℃ for staining, taken out after 30min and fixed in 10% formaldehyde, and photographed after 24 h. Calculating the infarct size by adopting an image analysis system, and accumulating the product of the infarct size and the thickness of each brain slice to obtain the infarct volume.

2.5 brain tissue Evans blue quantitation

10 rats were taken from each group and treated with 10% chloral hydrate (3.5 mL^{－ 1}) Under anesthesia, the left ventricle was perfused with saline until the effluent was colorless, and 2% Evans blue (1 mL/mouse) was injected from the right femoral vein. Accurately weighing the brain tissue by an electronic balance, putting the brain tissue into a test tube of middle size, respectively adding 3 mL of formamide, covering the test tube, incubating the test tube in a water bath box at 45 ℃ for 48 h, shaking up the test tube gently, centrifuging the test tube at 3000 r/min for 15min, taking the supernatant, measuring the absorbance (A) at lambda = 632 nm, and calculating the content of Evans blue in the brain tissue.

2.6 measurement of cytokine content

10 rats in each group are taken, sacrificed 24 h after operation, brain tissues are quickly stripped on an ice tray, washed by ice-cold PBS, sucked dry by filter paper, immediately weighed by an electronic balance to prepare 10 percent brain tissue homogenate, the homogenate is centrifuged for 15min at the speed of 4000 r/min at the temperature of 4 ℃, supernatant is taken, and the contents of IL-1 beta, TNF-alpha, IL-6 and IL-8 are detected at the wavelength of 492 nm by adopting a double-antibody sandwich ABC-ELISA method.

3. Results of the experiment

3.1 Effect on neurological symptoms in ischemia-reperfusion rats

Experiments show that after 2 hours of ischemia, rats in the model group are subjected to reperfusion for 22 hours, and hemiplegia-like symptoms are caused, mainly manifested by adduction of contralateral forelimb, shoulder pronation, reduction of forelimb muscle tension, reduction of shoulder resistance and inclination towards healthy side. The experimental result shows that the pharmaceutical composition can obviously improve the neurological symptoms of rats subjected to ischemia reperfusion, has obvious treatment effect on the symptoms of stroke and hemiplegia caused by ischemia reperfusion, and has better effect than the 1-3# control sample and the commercial panax notoginseng saponins control medicament.

2. Effect on cerebral infarction volume of ischemia-reperfusion rat

The experimental results show that the pharmaceutical composition can significantly reduce the cerebral infarction volume of an ischemia-reperfusion rat, and the effect is superior to that of a 1-3# control sample and a commercially available panax notoginseng saponins control drug.

3. Influence on Evans blue content in brain tissue of ischemia-reperfusion rat

Blood Brain Barrier (BBB) disruption is an important pathophysiological basis for cerebral ischemia-reperfusion brain injury. Evans blue series small molecules are common BBB indicators. Evans blue binds to plasma proteins and normally cannot pass the blood brain barrier. When cerebral ischemia reperfusion is carried out, BBB is damaged, the Evans blue content in the brain tissue is increased, and the damage degree of the blood brain barrier can be reflected by measuring the Evans blue content in the brain tissue.

The experimental results show that the pharmaceutical composition can significantly reduce the content of evans blue in the brain of an ischemia-reperfusion rat, and the pharmaceutical composition has a protective effect on the blood brain barrier of the ischemia-reperfusion rat, and the effect of the pharmaceutical composition is superior to that of a 1-3# control sample and a commercially available panax notoginseng saponins control drug.

And (4) conclusion:

the pharmaceutical composition can obviously improve the neurological symptoms of ischemia-reperfusion rats and has obvious treatment effect on the symptoms of stroke and hemiplegia caused by ischemia-reperfusion; the cerebral infarction volume of an ischemia-reperfusion rat can be remarkably reduced; the Evans blue content of the brain of the ischemia reperfusion rat can be obviously reduced, and the pharmaceutical composition has a protective effect on the blood brain barrier of the ischemia reperfusion rat. Experiments also show that the protective effect of the pharmaceutical composition on cerebral ischemia-reperfusion injury is better than that of commercial panax notoginseng saponins and other contrast drugs with the same dose. The research shows that the pharmaceutical composition can be used for preventing or treating ischemic cerebrovascular diseases such as ischemic cerebral apoplexy, cerebral infarction, cerebral hemiplegia and the like, and the curative effect of the pharmaceutical composition is superior to that of the existing panax notoginseng saponins.

Experimental example 3 study on protective action against rat retinal nerve injury

1. Materials and methods

1.1 drugs and reagents

Brilliant Cresyl violet (Cresyl Vio-let): the Shanghai pharmaceutical group chemical reagents, Inc.

1.2 Experimental instruments

An operation microscope: gx.ss.zz-3, shanghai medical optics instruments and plants;

ophthalmic microsurgical instruments: suzhou Mingren medical devices, Inc.;

nikon optical microscope: alphaphot-2 YS2 and YS100, supplied by Nikon corporation; electronic analytical balance: AB204-N, Mettler Toledo group); digital photography microscope: model BX51, available from OLYMPUS, Inc.

1.3 Experimental animals

SD rats, provided by the experimental animal center of Kunming medical university, are male and female, have the body weight of 200-240 g, are 8-12 weeks old, have no obvious neck distortion through examination, have transparent cornea, clear iris blood vessels, have equi-large circles such as pupils, and are sensitive to light reaction. After 3 days of acclimatization, continuously measuring intraocular pressure for 3 days, and removing the part with average intraocular pressure higher or lower than normal intraocular pressure interval (9-16 mmHg).

1.4 animal grouping and administration

SD rats 110, randomly divided into 11 groups: a normal control group, a model group, a No. 1 medicinal composition group, a No. 2 medicinal composition group, a No. 3 medicinal composition group, a No. 4 medicinal composition group, a No. 5 medicinal composition group, a No. 1 control sample group, a No. 2 control sample group, a No. 3 control sample group and a commercial Panax notoginsenosides group, wherein each group contains 10 panax notoginsenosides. Except for the normal control group, 100 rats were cauterized by Akira method to close the right episcleral vein, and a model of sustained ocular hypertension retinal nerve injury was made for the rats, and the experiment was carried out to observe that the ocular pressure stabilized at 30mm Hg or more for 10 days. Each drug group is treated for 1 month by intragastric administration 1 time a day, and the normal control group and the model group are administered with distilled water with the same amount by intragastric administration. Intraocular pressure was measured 1 time per week. After the experiment period of each group, retinas were taken for full-thickness plating, and cresol purple staining was used for RGCL neuron counting.

1.5 computer image analysis

Each to-be-detected retina slice is symmetrically drawn through the optic papilla, is divided into 4 quadrants including the part above the temples, the part below the temples, the part above the noses and the part below the noses, each quadrant retina is divided into 3 equal parts, namely 3 areas in the center, the middle and the periphery, 3 points are randomly taken from each subarea, the area of each point is 32500 mu m, and the retinal neuron cells of each point are counted and converted into the cell number (cell number/mm) of each square millimeter²) The mean of the cell densities at 3 points per region was taken as the neuron density for that region. Vascular endothelial cells in RGCL are readily distinguished by morphology, cells with densely stained nuclei and no Neisseria species are glial cells, neither of which is taken into RGCL neuronal counts

2 results of the experiment

The experimental results show that the pharmaceutical composition can obviously reduce the damage of the persistent ocular hypertension to the retinal nerve, has good protection and repair effects on the retinal nerve damage, and can be used as a therapeutic drug for retina or optic neuropathy.

EXAMPLE 4 acute toxicity test

1, materials and methods:

1.1 test drugs: # 5 pharmaceutical composition, prepared as in example 5.

1.2 control drugs: the quality of the panax notoginseng saponins sold in the market meets the requirements of 2015 edition of Chinese pharmacopoeia, and is purchased from Chinese medicine GmbH.

1.3 Experimental animals: 130 SPF-grade healthy ICR mice are 3-4 weeks old and have the weight of 18.0-22.0 g.

1.4 test methods: 130 ICR mice, randomized into 13 groups: blank control group, test drug 1-6 dosage group, control drug 1-6 dosage group, 10 each group, each male and female half. According to the results of the preliminary test, the test drug and the control drug were each administered at a dose of 550mg/kg body weight as the highest dose, and 5 doses were administered downward at an interdose ratio of 0.80, for a total of 6 dose groups. Each group of mice was fasted for 12 h before the experiment, and then were administered by intraperitoneal injection for 1 time, and a blank control group was administered by intraperitoneal injection of physiological saline of the same volume. After administration, feeding normally, and observing for 1 time every 15min within 2 h after administration; observing for 1 time every 30min within 2-4 h after the medicine is applied; observing for 1 time every 1 hour within 4-8 hours after the medicine is applied; observing for 1 time every 4 hours within 8-24 hours after the medicine is applied; after the 2 nd day, observe 1 time each day, weigh the weight, closely observe the dynamics of each mouse eating, drinking water condition and possible dynamics, abnormal muscle movement, external reaction, pupil change, abnormal secretion, abnormal stool and urine, eyeball protrusion, eyelid ptosis, abnormal respiration, skin color change and other toxic reaction and death condition in 14 d. If some mice die, the dead mice are dissected, and pathological changes of organs such as heart, liver, spleen, lung, kidney, brain, stomach, small intestine and the like are examined for 14 d observation. According to the death number and the administration dose of mice in each dose group, the LD50 values of the test drug and the control drug were calculated by the Bliss method.

2. experimental results

The test method for measuring LD50 by one-time intravenous injection of mice is adopted to observe and compare the acute toxicity reaction of the 5# pharmaceutical composition and the commercial panax notoginseng saponins. The result shows that the LD50 of the 5# pharmaceutical composition ICR mouse by single intravenous injection is 397mg/kg, and the LD50 of the commercial panax notoginseng saponins ICR mouse by single intravenous injection is 306mg/kg, which indicates that the half lethal dose of the 5# pharmaceutical composition is higher than that of the commercial panax notoginseng saponins.

The acute toxicity test was performed on the pharmaceutical compositions # 1-4 of examples 1-4 according to the above experimental protocol, and the results showed that the differences between groups were not statistically significant, similar to the results of the acute toxicity test on the pharmaceutical composition # 5 of example 5.

The experiments show that the pharmaceutical composition has extremely high medicinal safety, and the safety of the pharmaceutical composition is superior to that of the existing panax notoginseng saponins.

Experimental example 5 Experimental study on vascular irritation, hemolysis and hypersensitivity

1. Materials and instruments

1.1 drugs, reagents

1.1 drugs and reagents

The above drugs are dissolved with 0.9% NaCl injection for clinical application.

Bovine serum albumin: supplied by Genbase Bio-science Co.

Evans blue: shanghai such as Geji Biotech development Ltd.

1.2 Main instruments

TP1020 dehydrator, EG1160 embedding machine, RM 2235 electric microtome, HI1210 sheet spreading machine, HI1220 sheet baking machine, MLB biological microscope: leica instruments, Germany;

MR231 model bench high-speed refrigerated centrifuge: provided by France Jiean group;

t6 uv-vis spectrophotometer: supplied by Beijing general analysis instruments, Inc.

1.3 Experimental animals

New Zealand rabbits: the weight of the common grade is 2.0-2.5 kg for each half of male and female, and is provided by the Guangdong province medical experiment animal center.

Adult healthy ICR mice, weight 22~25 g, Guangdong province medical laboratory animal center provides.

2. Experimental methods

2.1 vascular irritation test

Selecting 70 New Zealand rabbits, and randomly dividing the New Zealand rabbits into 7 groups: the composition comprises a negative control group, a No. 1 medicinal composition group, a No. 2 medicinal composition group, a No. 3 medicinal composition group, a No. 4 medicinal composition group, a No. 5 medicinal composition group and a commercial Panax notoginsenosides group, wherein each group comprises 10 animals and males. Selecting a left ear marginal vein for intravenous drip, administering an equal amount of 0.9% NaCl injection to a negative control group, wherein the drip rate is about 50 drops per minute and qd, continuously administering for 7 d, observing the reaction condition of the injection part of the New Zealand rabbit by naked eyes before administration and 48 and 72 hours after the last administration every day, scoring according to the vascular stimulation reaction score standard (shown in table 1), calculating the average score of each part, and evaluating the stimulation reaction degree according to the vascular stimulation reaction evaluation standard (shown in table 2). Meanwhile, the general conditions, fur, feces, activities and the like of the New Zealand rabbits were observed. At 72 h after the last administration, 2 (female and male half) New Zealand rabbits per group were euthanized, the left (administration side) rabbit ear of the New Zealand rabbit was excised along the root of the ear, and 1 tissue was taken at each of the injection site, the proximal end of the injection site, and the distal end of the injection site for histopathological examination. The remaining 2 (male and female halves) new zealand rabbits in each group were observed for 14 days and then harvested for histopathological examination to see how reversible the stimulatory responses were.

TABLE 1 vascular irritation response score criteria

TABLE 2 evaluation criteria for vascular irritation response

2.2 in vitro hemolysis assay

Collecting blood of about 30mL of New Zealand rabbit heart, placing the blood into a triangular flask containing glass beads, shaking for 10 min, removing fibrinogen to obtain defibrinated blood, adding about 10 times of NaCl injection, shaking uniformly, centrifuging at 1200 r/min for 15min, removing supernatant, washing the precipitated red blood cells for 2-3 times by using the NaCl injection according to the method until the supernatant does not show red, and preparing the obtained red blood cells into 2% suspension by using the NaCl injection for later use. 80 clean test tubes are taken and numbered as No. 1-8 in sequence, and 10 test tubes are used in each number. 2% erythrocyte suspension, NaCl injection, purified water and liquid medicine were added in this order as shown in Table 3, mixed well and immediately incubated in a thermostatic water bath at 37. + -. 0.5 ℃. Observation after 3 h incubation: if the solution is clear red, no cells or a small amount of red blood cells remain at the tube bottom, indicating hemolysis; the erythrocytes all settled down, and the supernatant was colorless and clear, indicating no hemolysis. The solution has a brownish red or reddish brown flocculent precipitate, which does not disperse after shaking, indicating that the erythrocyte coagulation occurs. If the phenomenon of erythrocyte agglomeration exists, the agglomerate can be placed on a glass slide, 2 drops of NaCl solution are dripped on the edge of a cover glass, and the observation under a microscope shows that the agglomerated erythrocyte can be dissolved out and is pseudo-agglomerated, and the agglomerated erythrocyte can not be dissolved out and is true agglomerated. After observation, the supernatant was taken, and the A value of each tube was read on a spectrophotometer at 545 nm in the blank of purified water, and the hemolysis ratio of each test tube was calculated by the following formula:

hemolysis rate = (A t-a nc)/(a pc-a nc) × 100%. In the formula, A t represents the absorbance of the test tube, Anc represents the absorbance of the negative control tube (i.e., tube No. 7), and Apc represents the absorbance of the positive control tube (i.e., tube No. 8). If the hemolysis rate is > 5%, it indicates that hemolysis is present.

Sample adding table (ml) for in vitro hemolysis test

2.3 allergy test

80 adult healthy ICR mice, weighing 22-25 g, were randomized into 8 groups: a normal saline control group, a positive medicament bovine serum albumin control group, a No. 1 medicinal composition group, a No. 2 medicinal composition group, a No. 3 medicinal composition group, a No. 4 medicinal composition group, a No. 5 medicinal composition group and a commercially available panax notoginseng saponins group. Each of the above test substances contained 0.4% Evans blue, and each group of mice was injected with the corresponding test substance via tail vein. Behavioral changes and auricle blue staining of mice were observed 30min after dosing. The number of animals presenting with blue staining of auricle, the total number of ears with blue staining, and the area of blue staining of auricle were recorded for each group. The auricle bluing area (S) is the area of bluing/auricle area, and the value of bluing is evaluated according to the ranking criteria shown below. Then, each group of mice is killed after cervical vertebra is removed, double ears are taken and cut, 2mL of formamide is used for soaking for 2d, a 400-mesh sieve is used for filtering, and the supernatant is taken to measure the absorbance A at the position of 630 nm. The evans blue dye exudation amount (μ g) of both ears of each mouse was calculated from the evans blue standard curve. Calculating the positive rate (reaction rate for short) of the vascular reaction of each group according to the number of animals stained with auricle blue in each group; and calculating the ear blue staining rate according to the number of blue stained ears.

Grading index of blue dye

3. Results of the experiment

3.1 vascular irritation test

The experimental results show that the vascular irritation reaction value of the pharmaceutical composition provided by the invention to the intravenous drip of New Zealand rabbits is 0.11-0.23, the pharmaceutical composition belongs to a non-irritant grade, and the vascular irritation of the pharmaceutical composition is obviously lower than that of the panax notoginseng saponins sold in the market as a control medicament.

3.2 hemolytic test

The experimental results show that the pharmaceutical composition has no obvious hemolytic property, and the hemolytic rate of the pharmaceutical composition is obviously lower than that of the panax notoginseng saponins sold in the market as a control medicament.

3.3 hypersensitivity

The experimental results show that the degree of inflammatory reaction caused by anaphylaxis of the pharmaceutical composition is slight and is obviously lower than that of panax notoginseng saponins sold in the market as a control medicament.

And (4) conclusion: the experimental results show that the pharmaceutical composition has no obvious adverse reactions such as irritation, hemolytic property, anaphylaxis and the like, completely meets the requirement of medicinal safety, and has better safety evaluation indexes of various adverse reactions than the panax notoginseng saponins control drug.

Typical cases are:

1. zhangzhi, male, age 65, with diabetes for 7 years. From 5 months in 2014, the eyes are felt to be degraded, the right eye and the left eye of the vision are examined to be 0.5 and 0.1, microangioma can be seen in the retina, the fovea centralis reflects light and disappears, and the diabetic retinopathy is confirmed. After taking the 5# Xuesaitong soft capsule of the invention in the embodiment 18, 1 granule a day and 3 months, the examination shows that the vision of the right eye is 1.0, the vision of the left eye is 0.6, the vitreous opacity of the eyes is absorbed, and the blood stasis of the capillary is reduced. The follow-up visit is half a year without relapse.

2. Li A certain woman, age 58, with diabetes for 7 years, feels that binocular vision is degraded and the anterior shadow flies in front of the eyes since 2014 8. After examination, retinal vein is dilated, microangioma and hemorrhagic macula are existed, left eye vision is 0.3, right eye vision is 0.4, and diabetic retinopathy is confirmed. After taking the 4# Xuesaitong soft capsule of the embodiment 17 of the invention 1 granule a day and 3 months later, the flying shadow of the visual objects is reduced, the blood stasis of the capillary vessels is reduced, the vision of the left eye is recovered to 0.9, and the vision of the right eye is recovered to 1.0. The follow-up visit is half a year without relapse.

3. The 3# Xuesaitong soft capsule of the embodiment 16 of the invention is taken by students in the first, middle and third years, the load of the class school is large in daily life, the eyes are very tired after the students study in the daytime, the eyes are swollen in the daytime, the eyes are usually painful at night, the visual objects are blurred, the lacrimation and other conditions disappear after 1 month, and the visual fatigue condition is obviously improved.

4. In the year 35, the company investigators of a certain pharmaceutical company often use computers and mobile phones for work. From 2015 9, eyes were particularly fatigued, dry, itchy, painful, and blurred vision. The symptoms are not obviously improved by using eye drops such as ofloxacin. After the 2# Xuesaitong soft capsule of the embodiment 15 of the invention is taken 1 granule a day and 3 months, the symptoms of dry eyes, itching and pain disappear, and the vision is obviously improved. The follow-up visit is half a year without relapse.

Claims

1. The pharmaceutical composition for treating the thromboembolism is characterized by being prepared from the following components:

ginsenoside Rb254.6%

Ginsenoside Rb121.1%

Ginsenoside Rg 111.3%

Ginsenoside Rc 8.5%

Notoginsenoside R14.5%.

2. The pharmaceutical composition for treating the thromboembolism is characterized by being prepared from the following components:

ginsenoside Rb220.5%

Ginsenoside Rb147.8%

Ginsenoside Rg123.4%

Ginsenoside Rc 6.7%

11.6 percent of notoginsenoside R.

3. The pharmaceutical composition for treating the thromboembolism is characterized by being prepared from the following components:

ginsenoside Rb21.2%

Ginsenoside Rb114.6%

Ginsenoside Rg 155.7%

Ginsenoside Rc 19.3%

19.2 percent of notoginsenoside R.

4. The pharmaceutical composition for treating the thromboembolism is characterized by being prepared from the following components:

ginsenoside Rb221.1%

Ginsenoside Rb12.5%

Ginsenoside Rg 128.2%

Ginsenoside Rc 37.6%

Notoginsenoside R110.6%.

5. The pharmaceutical composition for treating the thromboembolism is characterized by being prepared from the following components:

ginsenoside Rb220.9%

Ginsenoside Rb12.7%

Ginsenoside Rg 121.5%

Ginsenoside Rc 10.2%

Notoginsenoside R144.7%.

6. A method for preparing the pharmaceutical composition for treating thromboembolism as claimed in any one of claims 1 to 5, which comprises the following steps:

step 1), weighing notoginsenoside R1, placing the notoginsenoside R1 in an ethanol solvent, and stirring for dissolving to obtain a solution A;

step 2), weighing ginsenoside Rb2, ginsenoside Rb1, ginsenoside Rg1 and ginsenoside Rc, stirring and dissolving in distilled water, heating to 50-70 ℃, adding the solution A while stirring, adding activated carbon for injection, heating to boil for 30-50 minutes, standing and cooling to room temperature, and filtering to remove the activated carbon;

7. The preparation method according to claim 6, wherein the mass ratio of the notoginsenoside R1 to the ethanol solvent in the step 1) is 0.05-5: 100, and the volume percentage concentration of the ethanol is 50-100%.

8. The preparation method according to claim 6, wherein the mass ratio of the four ginsenosides in step 2) to the activated carbon for injection and the distilled water is 0.1-10: 0.01-0.5: 100.

9. The preparation method according to claim 6, wherein the pre-freezing and cooling rate in the step 3) is 5-15 ℃/h.

10. A preparation of Xuesaitong medicine, which is characterized in that the preparation is prepared from the Xuesaitong medicine composition as claimed in any one of claims 1 to 5 and pharmaceutical auxiliary materials and is prepared into injections, tablets, oral liquid, capsules or dropping pills.

11. A soft capsule preparation of Xuesaitong, characterized in that it is prepared by using the pharmaceutical composition of any claim 1 to 5 as the effective component of the medicine, and adding proper auxiliary materials.

12. The soft capsule formulation of claim 11, wherein the soft capsule formulation comprises the following pharmaceutically active ingredients in mass percent:

ginsenoside Rb254.6%

Ginsenoside Rb121.1%

Ginsenoside Rg 111.3%

Ginsenoside Rc 8.5%

Notoginsenoside R14.5%.

13. The soft capsule formulation of claim 11, wherein the soft capsule formulation comprises the following pharmaceutically active ingredients in mass percent:

ginsenoside Rb220.5%

Ginsenoside Rb147.8%

Ginsenoside Rg123.4%

Ginsenoside Rc 6.7%

11.6 percent of notoginsenoside R.

14. The soft capsule formulation of claim 11, wherein the soft capsule formulation comprises the following pharmaceutically active ingredients in mass percent:

ginsenoside Rb21.2%

Ginsenoside Rb114.6%

Ginsenoside Rg 155.7%

Ginsenoside Rc 19.3%

19.2 percent of notoginsenoside R.

15. The soft capsule formulation of claim 11, wherein the soft capsule formulation comprises the following pharmaceutically active ingredients in mass percent:

ginsenoside Rb221.1%

Ginsenoside Rb12.5%

Ginsenoside Rg 128.2%

Ginsenoside Rc 37.6%

Notoginsenoside R110.6%.

16. The soft capsule formulation of claim 11, wherein the soft capsule formulation comprises the following pharmaceutically active ingredients in mass percent:

ginsenoside Rb220.9%

Ginsenoside Rb12.7%

Ginsenoside Rg 121.5%

Ginsenoside Rc 10.2%

Notoginsenoside R144.7%.

17. The soft capsule preparation according to claim 11, wherein the active ingredient of the drug according to any one of claims 12 to 16 is weighed, added to a solvent, stirred uniformly, placed in a pelleting press, pelleted with capsule skin, and dried to obtain the soft capsule preparation.

18. The soft capsule preparation of claim 17, wherein the solvent is any one or a mixture of any two or more of water, ethanol, ethylene glycol, propylene glycol, glycerol, tween, peanut oil, ginger oil, zanthoxylum oil, rose oil, rapeseed oil, peony seed oil, black peach oil, sunflower oil, patchouli oil, lard, beef tallow, mutton fat, fish oil, shark oil and snake oil in any proportion.

19. The soft capsule preparation of claim 17, wherein the capsule shell is prepared from a colloid, a solvent, a blending agent, hot-melt, a blend, a plastic, and a stabilizing agent.

20. The soft capsule formulation of claim 19, wherein the colloid from which the capsule shell is made is selected from the group consisting of animal gums and vegetable gums.

21. The soft capsule formulation of claim 20, wherein the animal glue is gelatin, bovine hide glue, dog hide glue or porcine hide glue.

22. The soft capsule formulation of claim 20, wherein the vegetable gum is carrageenan, guar gum, alginate gum, locust bean gum, konjac gum, or xanthan gum.

23. The soft capsule preparation according to claim 19, wherein the solvent for preparing the capsule shell is any one or a mixture of any two or more of water, ethanol, propanol, ethylene glycol, isopropanol, n-glycerol, isobutanol, t-butanol, n-butanol, and polyethylene glycol in any ratio.

24. The soft capsule formulation of claim 19, wherein the blending agent for preparing the capsule shell is a toning agent and an acid blending agent.

25. The soft capsule formulation according to claim 24, characterized in that the hueing agent is carmine, amaranth or olive green.

26. The method for preparing the soft capsule formulation for thromboembolism relieving as claimed in claim 24, wherein the acid regulator is citric acid, sodium citrate, carbonic acid, sodium carbonate, sodium bicarbonate, phosphoric acid, trisodium phosphate, disodium phosphate, monosodium phosphate, malic acid, sodium malate, lactic acid or sodium lactate.

27. The soft capsule formulation of claim 19, wherein the agent for preparing the capsule shell is a softening agent.

28. The soft capsule formulation of claim 27, wherein the softening agent is cellulose, methylcellulose, carboxymethylcellulose, hydroxymethylcellulose, beeswax, paraffin, stearic acid, or glycerol monostearate.

29. The application of the pharmaceutical composition for treating thromboembolism as claimed in any one of claims 1 to 5 in preparing medicaments for preventing and/or treating cardiovascular and cerebrovascular diseases.

30. The application of the pharmaceutical composition for treating thromboembolism as claimed in any one of claims 1 to 5 in preparing medicaments for preventing and/or treating ischemic cardiovascular and cerebrovascular diseases.

31. Use of the pharmaceutical composition for treating cerebral infarction according to any one of claims 1 to 5 in the preparation of a medicament for preventing and/or treating ischemic cerebral apoplexy, cerebral infarction and cerebral hemiplegia.

32. Use of the pharmaceutical composition for treating thromboembolism as claimed in any one of claims 1 to 5 in the preparation of a medicament for the prevention and/or treatment of retinopathy.

33. Use of the pharmaceutical composition for treating thromboembolism according to any one of claims 1 to 5 in the preparation of a medicament for preventing and/or treating asthenopia or visual deterioration caused by optic atrophy, retinitis pigmentosa or macular degeneration.

34. Use of the pharmaceutical composition for treating thromboembolism as claimed in any one of claims 1 to 5 in preparing a medicament for preventing and/or treating retinopathy caused by diabetes, cardiovascular diseases, cerebrovascular diseases and neurological diseases.